PMID- 16306383 TI - Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Insight into the diversity and evolution of the cryptomonad nucleomorph genome. AB - The cryptomonads are an enigmatic group of marine and freshwater unicellular algae that acquired their plastids through the engulfment and retention of a eukaryotic ("secondary") endosymbiont. Together with the chlorarachniophyte algae, the cryptomonads are unusual in that they have retained the nucleus of their endosymbiont in a miniaturized form called a nucleomorph. The nucleomorph genome of the cryptomonad Guillardia theta has been completely sequenced and with only three chromosomes and a total size of 551 kb, is a model of nuclear genome compaction. Using this genome as a reference, we have investigated the structure and content of nucleomorph genomes in a wide range of cryptomonad algae. In this study, we have sequenced nine new cryptomonad nucleomorph 18S ribosomal DNA (rDNA) genes and four heat shock protein 90 (hsp90) gene fragments, and using pulsed-field gel electrophoresis and Southern hybridizations, have obtained nucleomorph genome size estimates for nine different species. We also used long range polymerase chain reaction to obtain nucleomorph genomic fragments from Hanusia phi CCMP325 and Proteomonas sulcata CCMP704 that are syntenic with the subtelomeric region of nucleomorph chromosome I in G. theta. Our results indicate that (1) the presence of three chromosomes is a common feature of the nucleomorph genomes of these organisms, (2) nucleomorph genome size varies dramatically in the cryptomonads examined, (3) unidentified cryptomonad species CCMP1178 has the largest nucleomorph genome identified to date at approximately 845 kb, (4) nucleomorph genome size reductions appear to have occurred multiple times independently during cryptomonad evolution, (5) the relative positions of the 18S rDNA, ubc4, and hsp90 genes are conserved in three different cryptomonad genera, and (6) interchromosomal recombination appears to be rapidly changing the size and sequence of a repetitive subtelomeric region of the nucleomorph genome between the 18S rDNA and ubc4 loci. These results provide a glimpse into the genetic diversity of nucleomorph genomes in cryptomonads and set the stage for more comprehensive sequence-based studies in closely and distantly related taxa. PMID- 16306384 TI - Testing "species pair" hypotheses: evolutionary processes in the lichen-forming species complex Porpidia flavocoerulescens and Porpidia melinodes. AB - Pairs of taxa are commonly found in lichen-forming ascomycetes that differ primarily in their reproductive modes: one taxon reproduces sexually, the other vegetatively. The evolutionary processes underlying such "species pairs" are unknown. The species pair formed by Porpidia flavocoerulescens (sexual) and Porpidia melinodes (vegetative) was chosen to investigate four previously proposed hypotheses. These hypotheses posit that species pairs are either two monophyletic, independently evolving species with contrasting reproductive mode; a single outcrossing species polymorphic with regard to its reproductive modes; a sexual mother lineage frequently giving rise to asexual spin-offs; or a complex of cryptic species. The phylogenetic patterns observed within the species pair in the present study were analyzed using stringent hypothesis testing and visualizations of relationships and conflict based on tree and network reconstructions. DNA sequences at the three analyzed loci revealed the same four to five deeply divergent lineages. A detailed analysis of DNA-sequence variability revealed closely linked gene loci, but high levels of conflict within each of the gene fragments, as well as between observed genetic lineages. The observed patterns of phylogenetic relationships, linkage, and conflict are not congruent with any of the previously proposed species pair hypotheses. Rather, it is proposed that the observed results can be explained by conflicting reproductive and nutritional requirements imposed by an obligate symbiotic lifestyle. These interacting constraints produce recurring selective sweeps within predominantly vegetatively reproducing lineages and are the main forces that shape the evolution within the investigated species pair. PMID- 16306385 TI - The emerging role of the endocannabinoid system in endocrine regulation and energy balance. AB - During the last few years, the endocannabinoid system has emerged as a highly relevant topic in the scientific community. Many different regulatory actions have been attributed to endocannabinoids, and their involvement in several pathophysiological conditions is under intense scrutiny. Cannabinoid receptors, named CB1 receptor and CB2 receptor, first discovered as the molecular targets of the psychotropic component of the plant Cannabis sativa, participate in the physiological modulation of many central and peripheral functions. CB2 receptor is mainly expressed in immune cells, whereas CB1 receptor is the most abundant G protein-coupled receptor expressed in the brain. CB1 receptor is expressed in the hypothalamus and the pituitary gland, and its activation is known to modulate all the endocrine hypothalamic-peripheral endocrine axes. An increasing amount of data highlights the role of the system in the stress response by influencing the hypothalamic-pituitary-adrenal axis and in the control of reproduction by modifying gonadotropin release, fertility, and sexual behavior. The ability of the endocannabinoid system to control appetite, food intake, and energy balance has recently received great attention, particularly in the light of the different modes of action underlying these functions. The endocannabinoid system modulates rewarding properties of food by acting at specific mesolimbic areas in the brain. In the hypothalamus, CB1 receptor and endocannabinoids are integrated components of the networks controlling appetite and food intake. Interestingly, the endocannabinoid system was recently shown to control metabolic functions by acting on peripheral tissues, such as adipocytes, hepatocytes, the gastrointestinal tract, and, possibly, skeletal muscle. The relevance of the system is further strenghtened by the notion that drugs interfering with the activity of the endocannabinoid system are considered as promising candidates for the treatment of various diseases, including obesity. PMID- 16306387 TI - Modeling genome evolution with a diffusion approximation of a birth-and-death process. AB - MOTIVATION: In our previous studies, we developed discrete-space birth, death and innovation models (BDIMs) of genome evolution. These models explain the origin of the characteristic Pareto distribution of paralogous gene family sizes in genomes, and model parameters that provide for the evolution of these distributions within a realistic time frame have been identified. However, extracting the temporal dynamics of genome evolution from discrete-space BDIM was not technically feasible. We were interested in obtaining dynamic portraits of the genome evolution process by developing a diffusion approximation of BDIM. RESULTS: The diffusion version of BDIM belongs to a class of continuous-state models whose dynamics is described by the Fokker-Plank equation and the stationary solution could be any specified Pareto function. The diffusion models have time-dependent solutions of a special kind, namely, generalized self-similar solutions, which describe the transition from one stationary distribution of the system to another; this provides for the possibility of examining the temporal dynamics of genome evolution. Analysis of the generalized self-similar solutions of the diffusion BDIM reveals a biphasic curve of genome growth in which the initial, relatively short, self-accelerating phase is followed by a prolonged phase of slow deceleration. This evolutionary dynamics was observed both when genome growth started from zero and proceeded via innovation (a potential model of primordial evolution), and when evolution proceeded from one stationary state to another. In biological terms, this regime of evolution can be tentatively interpreted as a punctuated-equilibrium-like phenomenon whereby evolutionary transitions are accompanied by rapid gene amplification and innovation, followed by slow relaxation to a new stationary state. PMID- 16306388 TI - Incorporation of splice site probability models for non-canonical introns improves gene structure prediction in plants. AB - MOTIVATION: The vast majority of introns in protein-coding genes of higher eukaryotes have a GT dinucleotide at their 5'-terminus and an AG dinucleotide at their 3' end. About 1-2% of introns are non-canonical, with the most abundant subtype of non-canonical introns being characterized by GC and AG dinucleotides at their 5'- and 3'-termini, respectively. Most current gene prediction software, whether based on ab initio or spliced alignment approaches, does not include explicit models for non-canonical introns or may exclude their prediction altogether. With present amounts of genome and transcript data, it is now possible to apply statistical methodology to non-canonical splice site prediction. We pursued one such approach and describe the training and implementation of GC-donor splice site models for Arabidopsis and rice, with the goal of exploring whether specific modeling of non-canonical introns can enhance gene structure prediction accuracy. RESULTS: Our results indicate that the incorporation of non-canonical splice site models yields dramatic improvements in annotating genes containing GC-AG and AT-AC non-canonical introns. Comparison of models shows differences between monocot and dicot species, but also suggests GC intron-specific biases independent of taxonomic clade. We also present evidence that GC-AG introns occur preferentially in genes with atypically high exon counts. AVAILABILITY: Source code for the updated versions of GeneSeqer and SplicePredictor (distributed with the GeneSeqer code) isavailable at http://bioinformatics.iastate.edu/bioinformatics2go/gs/download.html. Web servers for Arabidopsis, rice and other plant species are accessible at http://www.plantgdb.org/PlantGDB-cgi/GeneSeqer/AtGDBgs.cgi, http://www.plantgdb.org/PlantGDB-cgi/GeneSeqer/OsGDBgs.cgi and http://www.plantgdb.org/PlantGDB-cgi/GeneSeqer/PlantGDBgs.cgi, respectively. A SplicePredictor web server is available at http://bioinformatics.iastate.edu/cgi bin/sp.cgi. Software to generate training data and parameterizations for Bayesian splice site models is available at http://gremlin1.gdcb.iastate.edu/~volker/SB05B/BSSM4GSQ/ PMID- 16306389 TI - The choice of optimal distance measure in genome-wide datasets. AB - MOTIVATION: Many types of genomic data are naturally represented as binary vectors. Numerous tasks in computational biology can be cast as analysis of relationships between these vectors, and the first step is, frequently, to compute their pairwise distance matrix. Many distance measures have been proposed in the literature, but there is no theory justifying the choice of distance measure. RESULTS: We examine the approaches to measuring distances between binary vectors and study the characteristic properties of various distance measures and their performance in several tasks of genome analysis. Most distance measures between binary vectors turn out to belong to a single parametric family, namely generalized average-based distance with different exponents. We show that descriptive statistics of distance distribution, such as skewness and kurtosis, can guide the appropriate choice of the exponent. On the contrary, the more familiar distance properties, such as metric and additivity, appear to have much less effect on the performance of distances. AVAILABILITY: R code GADIST and Supplementary material are available at http://research.stowers institute.org/bioinfo/ PMID- 16306390 TI - DNA assembly with gaps (Dawg): simulating sequence evolution. AB - MOTIVATION: Relationships amongst taxa are inferred from biological data using phylogenetic methods and procedures. Very few known phylogenies exist against which to test the accuracy of our inferences. Therefore, in the absence of biological data, simulated data must be used to test the accuracy of methods which produce these inferences. Researchers have limited or non-existent options for simulations useful for studying the impact of insertions, deletions, and alignments on phylogenetic accuracy. RESULTS: To satisfy this gap I have developed a new algorithm of indel formation and incorporated it into a new, flexible, and portable application for sequence simulation. The application, called Dawg, simulates phylogenetic evolution of DNA sequences in continuous time using the robust general time reversible model with gamma and invariant rate heterogeneity and a novel length-dependent model of indel formation. On completion, Dawg produces the true alignment of the simulated sequences. Unlike other applications, Dawg allows indel lengths to be explicitly distributed via a biologically realistic power law. Many options are available to allow users to customize their simulations and results. Because simulating with indels would be problematic if biologically realistic parameters could not be estimated, a script is provided with Dawg that can estimate the parameters of indel formation from sequence data. Dawg was applied to the sequences of four chloroplast trnK introns. It was used to parametrically bootstrap an estimation of the rate of indel formation for the phylogeny. Because Dawg can assist in parametric bootstrapping of sequence data it is useful beyond phylogenetics, such as studying alignment algorithms or parameters of molecular evolution. AVAILABILITY: Dawg 1.0.0 can be obtained at the following websites: http://www.genetics.uga.edu/sw/ or http://scit.us/dawg/. The package includes source code, example files, a brief manual and helper scripts. Binary distributions are available for Windows and Macintosh OS X. A development page for Dawg exists at http://scit.us/dawg/, with links to a Subversion repository, mailing lists and updated versions. PMID- 16306391 TI - Within-host evolution of CD8+-TL epitopes encoded by overlapping and non overlapping reading frames of simian immunodeficiency virus. AB - In order to understand the impact of overlapping reading frames on natural selection by host CD8+ T lymphocytes (CD8(+)-TL), we analyzed the pattern of nucleotide substitution in simian immunodeficiency virus (SIV) genomes sampled from populations at time of death in 35 rhesus monkeys. Both the mean number of nonsynonymous nucleotide substitutions per nonsynonymous site (d(N)) and the mean number of synonymous nucleotide substitutions per synonymous site (d(S)) were elevated in overlap regions in comparison to non-overlap regions. Mean d(N) exceeded mean d(S) in CD8(+)-TL epitopes restricted by the host's class I major histocompatibility complex molecules. This pattern, which is indicative of positive Darwinian selection favoring amino acid changes in these epitopes, was seen in both overlap and non-overlap regions; but mean d(N) was particularly elevated in restricted CD8(+)-TL epitopes encoded in overlap regions. Amino acid changes from the inoculum were defined as parallel if the same amino acid change occurred at the same site independently in two or more monkeys, and a surprisingly high proportion (71.9%) of observed amino acid changes throughout the SIV genome occurred in parallel in different monkeys. The proportion of parallel changes in restricted epitopes encoded by overlapping reading frames was still higher (80%), supporting the hypothesis that the interaction of positive selection and overlapping reading frames enhances the probability of convergent or parallel amino acid change. PMID- 16306392 TI - Four-helix bundle: a ubiquitous sensory module in prokaryotic signal transduction. AB - MOTIVATION: Transmembrane chemoreceptors in Escherichia coli utilize ligand binding domains for detecting various external signals. The structure of this domain in the E.coli aspartate receptor, Tar, is known and its signal transduction mechanism is under investigation. Current domain models for this important sensory module are inaccurate and, therefore, cannot reveal the distribution of this domain within the current genomic landscape. RESULTS: We carried out sensitive and exhaustive PSI-BLAST searches initiated with the sequence corresponding to a known structure of the four-helix, ligand-binding domain of the aspartate chemoreceptor. From the resulting sequences, we built a multiple sequence alignment for this domain family, which confirmed that the current TarH model is erroneous and fails to detect most of the domain homologs. In the process, we developed a technique that visualizes the secondary structure prediction of each protein sequence in order to improve the multiple sequence alignment. We found that the four-helix up-and-down bundle represents a large domain family and includes representatives of all major classes of prokaryotic signal transduction, namely histidine kinases, di-guanylate cyclases and chemotaxis receptors. PMID- 16306393 TI - Mining sequence annotation databanks for association patterns. AB - MOTIVATION: Millions of protein sequences currently being deposited to sequence databanks will never be annotated manually. Similarity-based annotation generated by automatic software pipelines unavoidably contains spurious assignments due to the imperfection of bioinformatics methods. Examples of such annotation errors include over- and underpredictions caused by the use of fixed recognition thresholds and incorrect annotations caused by transitivity based information transfer to unrelated proteins or transfer of errors already accumulated in databases. One of the most difficult and timely challenges in bioinformatics is the development of intelligent systems aimed at improving the quality of automatically generated annotation. A possible approach to this problem is to detect anomalies in annotation items based on association rule mining. RESULTS: We present the first large-scale analysis of association rules derived from two large protein annotation databases-Swiss-Prot and PEDANT-and reveal novel, previously unknown tendencies of rule strength distributions. Most of the rules are either very strong or very weak, with rules in the medium strength range being relatively infrequent. Based on dynamics of error correction in subsequent Swiss-Prot releases and on our own manual analysis we demonstrate that exceptions from strong rules are, indeed, significantly enriched in annotation errors and can be used to automatically flag them. We identify different strength dependencies of rules derived from different fields in Swiss-Prot. A compositional breakdown of association rules generated from PEDANT in terms of their constituent items indicates that most of the errors that can be corrected are related to gene functional roles. Swiss-Prot errors are usually caused by under-annotation owing to its conservative approach, whereas automatically generated PEDANT annotation suffers from over-annotation. AVAILABILITY: All data generated in this study are available for download and browsing at http://pedant.gsf.de/ARIA/index.htm. PMID- 16306394 TI - The adult mouse hippocampal progenitor is neurogenic but not a stem cell. AB - The aim of this investigation was to characterize the proliferative precursor cells in the adult mouse hippocampal region. Given that a very large number of new hippocampal cells are generated over the lifetime of an animal, it is predicted that a neural stem cell is ultimately responsible for maintaining this genesis. Although it is generally accepted that a proliferative precursor resides within the hippocampus, contradictory reports exist regarding the classification of this cell. Is it a true stem cell or a more limited progenitor? Using a strict functional definition of a neural stem cell and a number of in vitro assays, we report that the resident hippocampal precursor is a progenitor capable of proliferation and multipotential differentiation but is unable to self-renew and thus proliferate indefinitely. Furthermore, the mitogen FGF-2 stimulates proliferation of these cells to a greater extent than epidermal growth factor (EGF). In addition, we found that BDNF was essential for the production of neurons from the hippocampal progenitor cells, being required during proliferation to trigger neuronal fate. In contrast, a bona fide neural stem cell was identified in the lateral wall of the lateral ventricle surrounding the hippocampus. Interestingly, EGF proved to be the stronger mitogenic factor for this cell, which was clearly a different precursor from the resident hippocampal progenitor. These results suggest that the stem cell ultimately responsible for adult hippocampal neurogenesis resides outside the hippocampus, producing progenitor cells that migrate into the neurogenic zones and proliferate to produce new neurons and glia. PMID- 16306395 TI - Downstream regulatory element antagonist modulator regulates Ca2+ homeostasis and viability in cerebellar neurons. AB - The Na+/Ca2+ exchangers NCX1, NCX2, and NCX3 are vital for the control of cellular Ca2+ homeostasis. Here, we show that a doublet of downstream regulatory element sites in the promoter of the NCX3 gene mediates transcriptional repression of NCX3 by the Ca2+-modulated transcriptional repressor downstream regulatory element antagonist modulator (DREAM). Overexpression of a DREAM EF hand mutant insensitive to Ca2+ (EFmDREAM) in hippocampus and cerebellum of transgenic mice significantly reduced NCX3 mRNA and protein levels without modifying NCX1 and NCX2 expression. Cerebellar granules from EFmDREAM transgenic mice showed increased levels of cytosolic Ca2+ and were more vulnerable to increased Ca2+ influx after partial opening of voltage-gated plasma membrane Ca2+ channels induced by increasing K+ in the culture medium but survived better in the conditions of reduced Ca2+ influx prevailing in low extracellular K+. Overexpression of NCX3 in EFmDREAM transgenic granules using a lentiviral vector restored the normal survival response to high K+ observed in wild-type granules. Thus, the downregulation of the regulator of Ca2+ homeostasis NCX3 by Ca2+ regulated DREAM is a striking example of the autoregulatory property of the Ca2+ signal in neurons. PMID- 16306396 TI - Involvement of 5-HT1A receptors in prefrontal cortex in the modulation of dopaminergic activity: role in atypical antipsychotic action. AB - Atypical antipsychotics increase dopamine (DA) release in the medial prefrontal cortex (mPFC), an effect possibly involved in the superior effects of atypical versus classical antipsychotics on cognitive/negative symptoms. We examined the role of 5-HT1A receptors in the mPFC on the modulation of dopaminergic activity and the mesocortical DA release in vivo. The highly selective 5-HT1A agonist BAY x 3702 (BAY; 10-40 microg/kg, i.v.) increased the firing rate and burst firing of DA neurons in the ventral tegmental area (VTA) and DA release in the VTA and mPFC. The increase in DA release in both areas was potentiated by nomifensine coperfusion. The selective 5-HT1A antagonist WAY-100635 reversed the effects of BAY in both areas, and the changes in the VTA were prevented by frontocortical transection. The application of BAY in rat and mouse mPFC by reverse dialysis increased local extracellular DA at a low concentration (3 microM) and reduced it at a higher concentration (30 microM). Both effects disappeared in 5-HT1A knock out mice. In the presence of bicuculline, BAY reduced DA release at all concentrations. The atypical antipsychotics clozapine, olanzapine, and ziprasidone (but not haloperidol) enhanced DA release in the mPFC of wild-type but not 5-HT1A knock-out mice after systemic and local (clozapine and olanzapine) administration in the mPFC. Likewise, bicuculline coperfusion prevented the elevation of DA release produced by local clozapine or olanzapine application. These results suggest that the activation of mPFC 5-HT1A receptors enhances the activity of VTA DA neurons and mesocortical DA release. This mechanism may be involved in the elevation of extracellular DA produced by atypical antipsychotics. PMID- 16306397 TI - The suppressive field of neurons in lateral geniculate nucleus. AB - The responses of neurons in lateral geniculate nucleus (LGN) exhibit powerful suppressive phenomena such as contrast saturation, size tuning, and masking. These phenomena cannot be explained by the classical center-surround receptive field and have been ascribed to a variety of mechanisms, including feedback from cortex. We asked whether these phenomena might all be explained by a single mechanism, contrast gain control, which is inherited from retina and possibly strengthened in thalamus. We formalized an intuitive model of retinal contrast gain control that explicitly predicts gain as a function of local contrast. In the model, the output of the receptive field is divided by the output of a suppressive field, which computes the local root-mean-square contrast. The model provides good fits to LGN responses to a variety of stimuli; with a single set of parameters, it captures saturation, size tuning, and masking. It also correctly predicts that responses to small stimuli grow proportionally with contrast: were it not for the suppressive field, LGN responses would be linear. We characterized the suppressive field and found that it is similar in size to the surround of the classical receptive field (which is eight times larger than commonly estimated), it is not selective for stimulus orientation, and it responds to a wide range of frequencies, including very low spatial frequencies and high temporal frequencies. The latter property is hardly consistent with feedback from cortex. These measurements thoroughly describe the visual properties of contrast gain control in LGN and provide a parsimonious explanation for disparate suppressive phenomena. PMID- 16306398 TI - Quinoline and benzimidazole derivatives: candidate probes for in vivo imaging of tau pathology in Alzheimer's disease. AB - Neurofibrillary tangles (NFTs), neuropil threads, and neuritic elements of senile plaques predominantly comprise hyperphosphorylated tau protein and represent pathological characteristics of Alzheimer's disease (AD). These lesions occur before the presentation of clinical symptoms and correlate with the severity of dementia. In vivo detection of these lesions would thus prove useful for preclinical diagnosis of AD and for tracking disease progression. The present study introduces three novel compounds, 4-[2-(2-benzoimidazolyl)ethenyl]-N,N diethylbenzenamine (BF-126), 2-[(4-methylamino)phenyl]quinoline (BF-158), and 2 (4-aminophenyl)quinoline (BF-170), as candidate probes for in vivo imaging of tau pathology in the AD brain. When solutions of these compounds are injected intravenously into normal mice, these agents exhibit excellent brain uptake and rapid clearance from normal brain tissue. These compounds display relatively lower binding affinity to beta-amyloid fibrils and higher binding affinity to tau fibrils, compared with previously reported probe BF-168. In neuropathological examination using AD brain sections, BF-126, BF-158, and BF-170 clearly visualize NFTs, neuropil threads, and paired helical filament-type neuritis. Autoradiography using 11C-labeled BF-158 further demonstrated labeling of NFTs in AD brain sections. These findings suggest the potential usefulness of quinoline and benzimidazole derivatives for in vivo imaging of tau pathology in AD. PMID- 16306399 TI - A-type and T-type currents interact to produce a novel spike latency-voltage relationship in cerebellar stellate cells. AB - The modification of first-spike latencies by low-threshold and inactivating K+ currents (IA) have important implications in neuronal coding and synaptic integration. To date, cells in which first-spike latency characteristics have been analyzed have shown that increased hyperpolarization results in longer first spike latencies, producing a monotonic relationship between first-spike latency and membrane voltage. Previous work has established that cerebellar stellate cells express members of the Kv4 potassium channel subfamily, which underlie IA in many central neurons. Spike timing in stellate cells could be particularly important to cerebellar output, because the discharge of even single spikes can significantly delay spike discharge in postsynaptic Purkinje cells. In the present work, we studied the first-spike latency characteristics of stellate cells. We show that first-spike latency is nonmonotonic, such that intermediate levels of prehyperpolarization produce the longest spike latencies, whereas greater hyperpolarization or depolarization reduces spike latency. Moreover, the range of first-spike latency values can be substantial in spanning 20-128 ms with preceding membrane shifts of <10 mV. Using patch clamp and modeling, we illustrate that spike latency characteristics are the product of an interplay between IA and low-threshold calcium current (IT) that requires a steady-state difference in the inactivation parameters of the currents. Furthermore, we show that the unique first-spike latency characteristics of stellate cells have important implications for the integration of coincident IPSPs and EPSPs, such that inhibition can shift first-spike latency to differentially modulate the probability of firing. PMID- 16306400 TI - Energy inhibition elevates beta-secretase levels and activity and is potentially amyloidogenic in APP transgenic mice: possible early events in Alzheimer's disease pathogenesis. AB - Beta-secretase [beta-site amyloid precursor protein-cleaving enzyme 1 (BACE1)] is the key rate-limiting enzyme for the production of the beta-amyloid (Abeta) peptide involved in the pathogenesis of Alzheimer's disease (AD). BACE1 levels and activity are increased in AD brain and are likely to drive Abeta overproduction, but the cause of BACE1 elevation in AD is unknown. Interestingly, cerebral glucose metabolism and blood flow are both reduced in preclinical AD, suggesting that impaired energy production may be an early pathologic event in AD. To determine whether reduced energy metabolism would cause BACE1 elevation, we used pharmacological agents (insulin, 2-deoxyglucose, 3-nitropropionic acid, and kainic acid) to induce acute energy inhibition in C57/B6 wild-type and amyloid precursor protein (APP) transgenic (Tg2576) mice. Four hours after treatment, we observed that reduced energy production caused a approximately 150% increase of cerebral BACE1 levels compared with control. Although this was a modest increase, the effect was long-lasting, because levels of the BACE1 enzyme remained elevated for at least 7 d after a single dose of energy inhibitor. In Tg2576 mice, levels of the BACE1-cleaved APP ectodomain APPsbeta were also elevated and paralleled the BACE1 increase in both relative amount and duration. Importantly, cerebral Abeta40 levels in Tg2576 were increased to approximately 200% of control at 7 d after injection, demonstrating that energy inhibition was potentially amyloidogenic. These results support the hypothesis that impaired energy production in the brain may drive AD pathogenesis by elevating BACE1 levels and activity, which, in turn, lead to Abeta overproduction. This process may represent one of the earliest pathogenic events in AD. PMID- 16306401 TI - Choroid plexus megalin is involved in neuroprotection by serum insulin-like growth factor I. AB - The involvement of circulating insulin-like growth factor I (IGF-I) in the beneficial effects of physical exercise on the brain makes this abundant serum growth factor a physiologically relevant neuroprotective signal. However, the mechanisms underlying neuroprotection by serum IGF-I remain primarily unknown. Among many other neuroprotective actions, IGF-I enhances clearance of brain amyloid beta (Abeta) by modulating transport/production of Abeta carriers at the blood-brain interface in the choroid plexus. We found that physical exercise increases the levels of the choroid plexus endocytic receptor megalin/low-density lipoprotein receptor-related protein-2 (LRP2), a multicargo transporter known to participate in brain uptake of Abeta carriers. By manipulating choroid plexus megalin levels through viral-directed overexpression and RNA interference, we observed that megalin mediates IGF-I-induced clearance of Abeta and is involved in IGF-I transport into the brain. Through this dual role, megalin participates in the neuroprotective actions of IGF-I including prevention of tau hyperphosphorylation and maintenance of cognitive function in a variety of animal models of cognitive loss. Because we found that in normal aged animals, choroid plexus megalin/LRP2 is decreased, an attenuated IGF-I/megalin input may contribute to increased risk of neurodegeneration, including late-onset Alzheimer's disease. PMID- 16306402 TI - Aging-dependent and -independent modulation of associative learning behavior by insulin/insulin-like growth factor-1 signal in Caenorhabditis elegans. AB - Mutations in the insulin/IGF-1 neuroendocrine pathway extend lifespan and affect development, metabolism, and other biological processes in Caenorhabditis elegans and in other species. In addition, they may play a role in learning and memory. Investigation of the insulin/IGF-1 pathway may provide clues for the prevention of age-related declines in cognitive functions. Here, we examined the effects of the life-extending (Age) mutations, such as the age-1 (phosphatidylinositol 3-OH kinase) and daf-2 (insulin/IGF-1 receptor) mutations, on associative learning behavior called isothermal tracking. This thermotaxis learning behavior associates paired stimuli, temperature, and food. The age-1 mutation delayed the age-related decline of isothermal tracking, resulting in a 210% extension of the period that ensures it. The effect is dramatic compared with the extension of other physiological health spans. In addition, young adults of various Age mutants (age-1, daf-2, clk-1, and eat-2) showed increased consistency of temperature-food association, which may be caused by a common feature of the mutants, such as the secondary effects of life extension (i.e., enhanced maintenance of neural mechanisms). The age-1 and daf-2 mutants but not the other Age mutants showed an increase in temperature-starvation association through a different mechanism. Increased temperature-food association of the daf-2 mutant was dependent on neuronal Ca2+-sensor ncs-1, which modulates isothermal tracking in the AIY interneuron. Interestingly, mutations in the daf-7 TGFbeta gene, which functions in parallel to the insulin/IGF-1 pathway, caused deficits in acquisition of temperature-food and temperature-starvation association. This study highlights roles of the Age mutations in modulation of certain behavioral plasticity. PMID- 16306403 TI - Stoichiometry of the alpha9alpha10 nicotinic cholinergic receptor. AB - The alpha9 and alpha10 nicotinic cholinergic subunits assemble to form the receptor that mediates synaptic transmission between efferent olivocochlear fibers and hair cells of the cochlea. They are the latest vertebrate nicotinic cholinergic receptor (nAChR) subunits that have been cloned, and their identification has established a distant early divergent branch within the nAChR gene family. The alpha10 subunit serves as a "structural" component leading to heteromeric alpha9alpha10 nAChRs with distinct properties. We now have probed the stoichiometry of recombinant alpha9alpha10 nAChRs expressed in Xenopus oocytes. We have made use of the analysis of the population of receptors assembled from a wild-type subunit and its partner alpha9 or alpha10 subunit bearing a reporter mutation of a valine to threonine at position 13' of the second transmembrane domain (TM2). Because the mutation increased the sensitivity of the receptor for acetylcholine (ACh) but mutations at different subunits were not equivalent, the number of alpha9 and alpha10 subunits could be inferred from the number of components in compound concentration-response curves to ACh. The results were confirmed via the analysis of the effects of a mutation to threonine at position 17' of TM2. Because at this position the mutations at different subunits were equivalent, the stoichiometry was inferred directly from the shifts in the ACh EC50 values. We conclude that the recombinant alpha9alpha10 receptor is a pentamer with a (alpha9)2(alpha10)3 stoichiometry. PMID- 16306404 TI - Neural activity controls the synaptic accumulation of alpha-synuclein. AB - The presynaptic protein alpha-synuclein has a central role in Parkinson's disease (PD). However, the mechanism by which the protein contributes to neurodegeneration and its normal function remain unknown. Alpha-synuclein localizes to the nerve terminal and interacts with artificial membranes in vitro but binds weakly to native brain membranes. To characterize the membrane association of alpha-synuclein in living neurons, we used fluorescence recovery after photobleaching. Despite its enrichment at the synapse, alpha-synuclein is highly mobile, with rapid exchange between adjacent synapses. In addition, we find that alpha-synuclein disperses from the nerve terminal in response to neural activity. Dispersion depends on exocytosis, but unlike other synaptic vesicle proteins, alpha-synuclein dissociates from the synaptic vesicle membrane after fusion. Furthermore, the dispersion of alpha-synuclein is graded with respect to stimulus intensity. Neural activity thus controls the normal function of alpha synuclein at the nerve terminal and may influence its role in PD. PMID- 16306405 TI - Phorbol esters target the activity-dependent recycling pool and spare spontaneous vesicle recycling. AB - Using electrophysiology and styryl dye imaging, we studied the effect of phorbol 12-myristate 13-acetate (PMA) on activity-dependent and spontaneous vesicle recycling. In electrophysiological experiments, we found that the PMA effect depended on the maturational state of the synapses. Spontaneous neurotransmitter release from nascent synapses without a functional readily releasable pool (RRP) was unresponsive to PMA application. In contrast, mature synapses responded robustly to PMA application, consistent with previous studies. Using styryl dye imaging, we found that there was a PMA-dependent increase in the size of the RRP when PMA was present before, during, or after activity-dependent dye uptake, suggesting that this effect involves an increase in the population of the RRP by vesicles recruited from the reserve pool. Additionally, we found that when PMA was present during spontaneous dye uptake, there was an increase in dye labeling, and these additional dye-loaded vesicles showed rapid destaining in response to strong stimulation and were also releasable by hypertonic sucrose. In contrast, these observations were not reproducible when PMA treatment was performed after spontaneous dye uptake and extracellular dye washout. Together, these findings suggest that the increased spontaneous neurotransmission in the presence of PMA was attributable to release of vesicles from the RRP rather than an effect of PMA on the spontaneously recycling pool. Thus, the phorbol esters selectively regulate the activity-dependent pool of vesicles, indicating that priming mechanisms that prepare vesicles for fusion, which are targeted by phorbol esters, are different for the spontaneous and evoked forms of fusion. PMID- 16306406 TI - The small GTPase Rab7 controls the endosomal trafficking and neuritogenic signaling of the nerve growth factor receptor TrkA. AB - Nerve growth factor (NGF) and its TrkA receptor exert important bioactivities on neuronal cells such as promoting survival and neurite outgrowth. Activated TrkA receptors are not only localized on the cell surface but also in signaling endosomes, and internalized TrkA receptors are important for the mediation of neurite outgrowth. The regulation of the endosomal trafficking of TrkA is so far unknown. Because the endosome-associated GTPase Rab7 coimmunoprecipitated with TrkA, we examined whether the endosomal trafficking of TrkA might be under the control of Rab7. Inhibiting Rab7 by expression of a green fluorescent protein tagged, dominant-negative Rab7 variant resulted in endosomal accumulation of TrkA and pronounced enhancement of TrkA signaling in response to limited stimulations with NGF, such as increased activation of Erk1/2 (extracellular signal-regulated kinase 1/2), neurite outgrowth, and expression of GAP-43 (growth-associated protein 43). Our studies show that the endosomal GTPase Rab7 controls the endosomal trafficking and neurite outgrowth signaling of TrkA. Because mutations of Rab7 are found in patients suffering from hereditary polyneuropathies, dysfunction of Rab7 might contribute to neurodegenerative conditions by affecting the trafficking of neurotrophins. Moreover, strategies aimed at controlling Rab7 activity might be useful for the treatment of neurodegenerative diseases. PMID- 16306407 TI - Emerging patterns of neuronal responses in supplementary and primary motor areas during sensorimotor adaptation. AB - Acquisition and retention of sensorimotor skills have been extensively investigated psychophysically, but little is known about the underlying neuronal mechanisms. Here we examine the evolution of neural activity associated with adaptation to new kinematic tasks in two cortical areas: the caudal supplementary motor area (SMA proper), and the primary motor cortex (MI). We investigate the hypothesis that adaptation starts at premotor areas, i.e., higher in the hierarchy of computation, until a stable representation is formed in primary areas. In accordance with previous studies, we found that adaptation can be characterized by two phases: an early phase that is accompanied by fast and substantial reduction of errors, followed by a late phase with slower and more moderate improvements in behavior. We used unsupervised clustering to separate the activity of the single cells into groups of cells with similar response patterns, under the assumption that each such subpopulation forms a functional unit. We specifically observed the number of clusters in each cortical area during early and late phases of the adaptation and found that the number of clusters is higher in the SMA during early phases of adaptation. In contrast, a higher number of clusters was observed in MI only during late phases. Our results suggest a new approach to analyze responses of large populations of neurons and use it to show a hierarchy of dynamic reorganization of functional groups during adaptation. PMID- 16306408 TI - Age-related changes in the inhibitory response properties of dorsal cochlear nucleus output neurons: role of inhibitory inputs. AB - Age-related hearing loss frequently results in a loss in the ability to discriminate speech signals, especially in noise. This is attributable, in part, to a loss in temporal resolving power and ability to adjust dynamic range. Circuits in the adult dorsal cochlear nucleus (DCN) have been shown to preserve signal in background noise. Fusiform cells, major DCN output neurons, receive focused glycinergic inputs from tonotopically aligned vertical cells that also project to the ventral cochlear nucleus. Glycine-mediated inhibition onto fusiform cells results in decreased tone-evoked activity as intensity is increased at frequencies adjacent to characteristic frequency (CF). DCN output is thus shaped by glycinergic inhibition, which can be readily assessed in recordings from fusiform cells. Previous DCN studies suggest an age-related loss of markers for glycinergic neurotransmission. The present study postulated that response properties of aged fusiform cells would show a loss of inhibition, resembling conditions observed with glycine receptor blockade. The functional impact of aging was examined by comparing response properties from units meeting fusiform-cell criteria in young and aged rats. Fusiform cells in aged animals displayed significantly higher maximum discharge rates to CF tones than those recorded from young-adult animals. Fusiform cells of aged rats displayed significantly fewer nonmonotonic CF rate-level functions and an age-related change in temporal response properties. These findings are consistent with an age related loss of glycinergic input, likely from vertical cells, and with findings from other sensory aging studies suggesting a selective age-related decrement in inhibitory amino acid neurotransmitter function. PMID- 16306409 TI - Intraneuronal beta-amyloid expression downregulates the Akt survival pathway and blunts the stress response. AB - Early events in Alzheimer's disease (AD) pathogenesis implicate the accumulation of beta-amyloid (Abeta) peptide inside neurons in vulnerable brain regions. However, little is known about the consequences of intraneuronal Abeta on signaling mechanisms. Here, we demonstrate, using an inducible viral vector system to drive intracellular expression of Abeta42 peptide in primary neuronal cultures, that this accumulation results in the inhibition of the Akt survival signaling pathway. Induction of intraneuronal Abeta42 expression leads to a sequential decrease in levels of phospho-Akt, increase in activation of glycogen synthase kinase-3beta, and apoptosis. Downregulation of Akt also paralleled intracellular Abeta accumulation in vivo in the Tg2576 AD mouse model. Overexpression of constitutively active Akt reversed the toxic effects of Abeta through a mechanism involving the induction of heat shock proteins (Hsps). We used a small-interfering RNA approach to explore the possibility of a link between Akt activity and Hsp70 expression and concluded that neuroprotection by Akt could be mediated through downstream induction of Hsp70 expression. These results suggest that the early dysfunction associated with intraneuronal Abeta accumulation in AD involve the associated impairments of Akt signaling and suppression of the stress response. PMID- 16306410 TI - Upregulation of the voltage-gated sodium channel beta2 subunit in neuropathic pain models: characterization of expression in injured and non-injured primary sensory neurons. AB - The development of abnormal primary sensory neuron excitability and neuropathic pain symptoms after peripheral nerve injury is associated with altered expression of voltage-gated sodium channels (VGSCs) and a modification of sodium currents. To investigate whether the beta2 subunit of VGSCs participates in the generation of neuropathic pain, we used the spared nerve injury (SNI) model in rats to examine beta2 subunit expression in selectively injured (tibial and common peroneal nerves) and uninjured (sural nerve) afferents. Three days after SNI, immunohistochemistry and Western blot analysis reveal an increase in the beta2 subunit in both the cell body and peripheral axons of injured neurons. The increase persists for >4 weeks, although beta2 subunit mRNA measured by real-time reverse transcription-PCR and in situ hybridization remains unchanged. Although injured neurons show the most marked upregulation,beta2 subunit expression is also increased in neighboring non-injured neurons and a similar pattern of changes appears in the spinal nerve ligation model of neuropathic pain. That increased beta2 subunit expression in sensory neurons after nerve injury is functionally significant, as demonstrated by our finding that the development of mechanical allodynia-like behavior in the SNI model is attenuated in beta2 subunit null mutant mice. Through its role in regulating the density of mature VGSC complexes in the plasma membrane and modulating channel gating, the beta2 subunit may play a key role in the development of ectopic activity in injured and non-injured sensory afferents and, thereby, neuropathic pain. PMID- 16306411 TI - The mechanosensitivity of mouse colon afferent fibers and their sensitization by inflammatory mediators require transient receptor potential vanilloid 1 and acid sensing ion channel 3. AB - Mechanical hypersensitivity of the colon underlies in part the chronic abdominal pain experienced by patients with irritable bowel syndrome, yet the molecules that confer mechanosensitivity to colon sensory neurons and their contribution to visceral pain are unknown. We tested the hypothesis that transient receptor potential vanilloid 1 (TRPV1) and acid-sensing ion channel 3 (ASIC3) are peripheral mechanosensors in colon afferent neuronal fibers that mediate visceral nociceptive behavior in mice. Visceral nociception, modeled by the visceromotor response to colorectal distension, and colon afferent fiber mechanosensitivity were assessed in control (C57BL/6) mice and two congenic knock-out mouse strains with deletions of either TRPV1 or ASIC3. Phasic colon distension (15-60 mmHg) produced graded behavioral responses in all three mouse strains. However, both TRPV1 and ASIC3 knock-out mice were significantly less sensitive to distension, with an average response magnitude only 58 and 50% of controls, respectively. The behavioral deficits observed in both strains of knock-out mice were associated with a significant and selective reduction in afferent fiber sensitivity to circumferential stretch of the colon, an effect that was mimicked in control preparations by pretreatment with capsazepine, a TRPV1 antagonist, but not amiloride, a nonselective ASIC antagonist (both 500 microM). In addition, whereas stretch-evoked afferent fiber responses were enhanced by chemical inflammatory mediators in control mice, this effect was differentially impaired in both knock out mouse strains. These results demonstrate a peripheral mechanosensory role for TRPV1 and ASIC3 in the mouse colon that contributes to nociceptive behavior and possibly peripheral sensitization during tissue insult. PMID- 16306412 TI - Spatiotemporal gating of sensory inputs in thalamus during quiescent and activated states. AB - The main role of the thalamus is to relay sensory inputs to the neocortex according to the regulations dictated by behavioral state. Hence, changes in behavioral state are likely to transform the temporal and spatial properties of thalamocortical receptive fields. We compared the receptive fields of single cells in the ventroposterior medial thalamus (VPM) of urethane-anesthetized rats during quiescent states and during aroused (activated) states. During quiescent states, VPM cells respond to stimulation of a principal whisker (PW) and may respond modestly to one or a few adjacent whiskers (AWs). During either generalized forebrain activation or selective thalamic activation caused by carbachol infusion in the VPM, the responses to AWs enhance so that VPM receptive fields become much larger. Such enlargement is not observed at the level of the principal trigeminal nucleus, indicating that it originates within the thalamus. Interestingly, despite the increase in AW responses during activation, simultaneous deflection of the PW and AWs produced VPM responses that resembled the PW response, as if the AWs were not stimulated. This nonlinear summation of sensory responses was present during both quiescent and activated states. In conclusion, the thalamus suppresses the excitatory surround (AWs) of the receptive field during quiescent states and enlarges this surround during arousal. But, thalamocortical cells represent only the center (PW) of the receptive field when the center (PW) and surround (AWs) are stimulated simultaneously. PMID- 16306413 TI - Prediction and decoding of retinal ganglion cell responses with a probabilistic spiking model. AB - Sensory encoding in spiking neurons depends on both the integration of sensory inputs and the intrinsic dynamics and variability of spike generation. We show that the stimulus selectivity, reliability, and timing precision of primate retinal ganglion cell (RGC) light responses can be reproduced accurately with a simple model consisting of a leaky integrate-and-fire spike generator driven by a linearly filtered stimulus, a postspike current, and a Gaussian noise current. We fit model parameters for individual RGCs by maximizing the likelihood of observed spike responses to a stochastic visual stimulus. Although compact, the fitted model predicts the detailed time structure of responses to novel stimuli, accurately capturing the interaction between the spiking history and sensory stimulus selectivity. The model also accounts for the variability in responses to repeated stimuli, even when fit to data from a single (nonrepeating) stimulus sequence. Finally, the model can be used to derive an explicit, maximum likelihood decoding rule for neural spike trains, thus providing a tool for assessing the limitations that spiking variability imposes on sensory performance. PMID- 16306414 TI - Enhanced susceptibility to kainate-induced seizures, neuronal apoptosis, and death in mice lacking gangliotetraose gangliosides: protection with LIGA 20, a membrane-permeant analog of GM1. AB - Knock-out (KO) mice lacking gangliotetraose gangliosides attributable to disruption of the gene for GM2/GD2 synthase [GalNAcT (UDP-N acetylgalactosamine:GM3/GD3 beta-1,4-N-acetylgalactosaminyltransferase; EC 2.4.1.92 [EC])] are revealing key neural functions for the complex gangliosides of brain. This study has found such animals to be highly susceptible to kainic acid (KA)-induced seizures in terms of both seizure severity and duration. Intraperitoneal injection of 25 mg/kg KA produced status epilepticus for approximately 200 min in normal mice or heterozygotes and more than four times longer in the KO mice. The latter group suffered approximately 30% mortality, which increased to approximately 75% at dosage of 30 mg/kg KA, compared with 10 14% for the other two genotypes at the latter dosage. Nissl staining and terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling assay revealed substantial deterioration of pyramidal neurons attributable to apoptosis in the KO hippocampus, especially the CA3 region. Seizure activity in the KO mouse was only moderately diminished by intraperitoneal injection of GM1 ganglioside, whereas LIGA 20, a semisynthetic analog of GM1, substantially reduced both seizure severity and cell damage. The potency of LIGA 20 was correlated with its enhanced membrane permeability (compared with GM1), as seen in the increased uptake of [3H]LIGA 20 into the subcellular fractions of brain including cell nuclei. The latter finding is consonant with LIGA 20-induced restoration of the Na+/Ca2+ exchanger located at the inner membrane of the nuclear envelope in KO mice, an exchanger dependent on tight association with GM1 or its analog for optimal activity. These results point to a neuroprotective role for GM1 and its associated exchanger in the nucleus, based on regulation of Ca2+ flux between nucleoplasm and nuclear envelope. PMID- 16306415 TI - Attention modulates the responses of simple cells in monkey primary visual cortex. AB - Spatial attention has long been postulated to act as a spotlight that increases the salience of visual stimuli at the attended location. We examined the effects of attention on the receptive fields of simple cells in primary visual cortex (V1) by training macaque monkeys to perform a task with two modes. In the attended mode, the stimuli relevant to the animal's task overlay the receptive field of the neuron being recorded. In the unattended mode, the animal was cued to attend to stimuli outside the receptive field of that neuron. The relevant stimulus, a colored pixel, was briefly presented within a white-noise stimulus, a flickering grid of black and white pixels. The receptive fields of the neurons were mapped by correlating spikes with the white-noise stimulus in both attended and unattended modes. We found that attention could cause significant modulation of the visually evoked response despite an absence of significant effects on the overall firing rates. On further examination of the relationship between the strength of the visual stimulation and the firing rate, we found that attention appears to cause multiplicative scaling of the visually evoked responses of simple cells, demonstrating that attention reaches back to the initial stages of visual cortical processing. PMID- 16306416 TI - Single synaptic vesicle tracking in individual hippocampal boutons at rest and during synaptic activity. AB - How synaptic vesicles move within central nervous synapses to their docking sites at the plasma membrane is widely discussed in synaptic physiology. This question is especially difficult to investigate in the small hippocampal boutons, which themselves can slowly move during observation in primary cell culture. Here, we describe a single particle tracking method using dual fluorescent dye labels that enabled us to visualize the movements of a single vesicle and the respective synaptic bouton simultaneously during resting conditions and stimulation. We found vesicle mobility to be very low in the absence of stimulation, in line with previous studies. Interestingly, mobility was also found to be low during synaptic activity. We found that vesicles labeled preferentially via early, late, and spontaneous endocytotic mechanisms behaved similarly at rest and during stimulation. PMID- 16306417 TI - Reversal of maternal programming of stress responses in adult offspring through methyl supplementation: altering epigenetic marking later in life. AB - Stress responses in the adult rat are programmed early in life by maternal care and associated with epigenomic marking of the hippocampal exon 1(7) glucocorticoid receptor (GR) promoter. To examine whether such epigenetic programming is reversible in adult life, we centrally infused the adult offspring with the essential amino acid L-methionine, a precursor to S-adenosyl-methionine that serves as the donor of methyl groups for DNA methylation. Here we report that methionine infusion reverses the effect of maternal behavior on DNA methylation, nerve growth factor-inducible protein-A binding to the exon 1(7) promoter, GR expression, and hypothalamic-pituitary-adrenal and behavioral responses to stress, suggesting a causal relationship among epigenomic state, GR expression, and stress responses in the adult offspring. These results demonstrate that, despite the inherent stability of the epigenomic marks established early in life through behavioral programming, they are potentially reversible in the adult brain. PMID- 16306418 TI - Separate face and body selectivity on the fusiform gyrus. AB - Recent reports of a high response to bodies in the fusiform face area (FFA) challenge the idea that the FFA is exclusively selective for face stimuli. We examined this claim by conducting a functional magnetic resonance imaging experiment at both standard (3.125 x 3.125 x 4.0 mm) and high resolution (1.4 x 1.4 x 2.0 mm). In both experiments, regions of interest (ROIs) were defined using data from blocked localizer runs. Within each ROI, we measured the mean peak response to a variety of stimulus types in independent data from a subsequent event-related experiment. Our localizer scans identified a fusiform body area (FBA), a body-selective region reported recently by Peelen and Downing (2005) that is anatomically distinct from the extrastriate body area. The FBA overlapped with and was adjacent to the FFA in all but two participants. Selectivity of the FFA to faces and FBA to bodies was stronger for the high-resolution scans, as expected from the reduction in partial volume effects. When new ROIs were constructed for the high-resolution experiment by omitting the voxels showing overlapping selectivity for both bodies and faces in the localizer scans, the resulting FFA* ROI showed no response above control objects for body stimuli, and the FBA* ROI showed no response above control objects for face stimuli. These results demonstrate strong selectivities in distinct but adjacent regions in the fusiform gyrus for only faces in one region (the FFA*) and only bodies in the other (the FBA*). PMID- 16306419 TI - Selection of nonapoptotic spermatozoa as a new tool for enhancing assisted reproduction outcomes: an in vitro model. AB - Magnetic cell sorting (MACS) using annexin V-conjugated microbeads eliminates apoptotic spermatozoa based on the externalization of phosphatidylserine residues. The procedure delivers two sperm fractions: annexin V-negative (nonapoptotic) and annexin V-positive (apoptotic). Our aim was to determine whether the sperm fertilizing potential can be improved by selecting a nonapoptotic fraction using MACS. Semen samples (n = 35) were subjected to separation on a density gradient followed by MACS. Extent of apoptosis was assessed by measuring levels of activated caspase 3 using fluorescein-labeled inhibitors of caspase, alterations in mitochondrial membrane potential (MMP) using a lipophilic cationic dye, and DNA fragmentation using terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labeling assay. The sperm fertilization potential was assessed using hamster oocyte penetration assay and hamster oocyte-intracytoplasmic sperm injection (ICSI). Annexin V negative sperm displayed superior quality in terms of high motility, low caspase 3 activation, MMP integrity, and small extent of DNA fragmentation. Annexin V negative sperm demonstrated higher oocyte penetration capacity but comparable sperm chromatin decondensation (SCD) following ICSI. Conversely, the annexin V positive sperm presented with poor quality and fertilization potential. The oocyte penetration rate was negatively correlated with apoptotic marker expression, whereas SCD following ICSI was only associated with apoptosis on sperm-damaged membranes. We conclude that apoptosis appears to impact sperm oocyte penetration rate; however, it does not seem to affect early stages of fertilization such as SCD in spermatozoa of healthy donors. The selection of nonapoptotic sperm by MACS may be used to enhance results of in vitro fertilization by increasing sperm-oocyte penetration. PMID- 16306420 TI - Anchorage-independent growth of mouse male germline stem cells in vitro. AB - Spermatogenesis originates from a small number of spermatogonial stem cells that reside on the basement membrane and undergo self-renewal division to support spermatogenesis throughout the life of adult animals. Although the recent development of a technique to culture spermatogonial stem cells allowed reproduction of self-renewal division in vitro, much remains unknown about how spermatogonial stem cells are regulated. In this study, we found that spermatogonial stem cells could be cultured in an anchorage-independent manner, which is characteristic of stem cells from other types of self-renewing tissues. Although the cultured cells grew slowly (doubling time, approximately 4.7 days), they expressed markers of spermatogonia, and grew exponentially for at least 5 months to achieve 1.5 x 10(10) -fold expansion. The cultured cells underwent spermatogenesis following transplantation into the seminiferous tubules of infertile animals and fertile offspring were obtained by microinsemination of germ cells that had developed within the testes of recipients of the cultured cells. These results indicate that spermatogonial stem cells can undergo anchorage-independent, self-renewal division, and suggest that stem cells have the common property to survive and proliferate in the absence of exogenous substrata. PMID- 16306421 TI - Immunocontraceptive effects on female rabbits infected with recombinant myxoma virus expressing rabbit ZP2 or ZP3. AB - Recombinant myxoma viruses expressing rabbit zona pellucida 2 (rZP2) or rabbit zona pellucida 3 (rZP3) glycoproteins were constructed and tested in domestic rabbits to assess their potential to induce autoimmune infertility. The recombinant virus expressing rZP2 had no effect on fertility or ovarian histology, despite all animals developing antibodies against the rZP2 antigen. However, recombinant viruses expressing rZP3 induced infertility in 70% of animals at the first breeding. Serum antibodies were relatively short-lived, but antibody was bound to zona pellucida of all rabbits from Day 10 onward. There was no obvious correlation between infertility and rZP3 antibody titer. There was a transient inflammatory response in the ovaries of rZP3-immunized rabbits at Day 15 but no T-cell response to rZP3 could be detected at any time. Dysfunctional follicular formation was present in ovaries from rabbits infected with rZP3 expressing viruses 15-40 days postinfection but this had disappeared at later time points. A recombinant myxoma virus expressing a modified rZP3 antigen with the C-terminal hydrophobic putative anchor sequence deleted was also tested. This virus did not induce either infertility or an antibody response against the zona pellucida. Thus, the context of antigen presentation was crucial for an autoimmune response. PMID- 16306422 TI - Clonal amniotic fluid-derived stem cells express characteristics of both mesenchymal and neural stem cells. AB - Recent evidence has shown that amniotic fluid may be a novel source of fetal stem cells for therapeutic transplantation. We previously developed a two-stage culture protocol to isolate a population of amniotic fluid-derived mesenchymal stem cells (AFMSCs) from second-trimester amniocentesis. AFMSCs maintain the capacity to differentiate into multiple mesenchymal lineages and neuron-like cells. It is unclear whether amniotic fluid contains heterogeneous populations of stem cells or a subpopulation of primitive stem cells that are similar to marrow stromal cells showing the behavior of neural progenitors. In this study, we showed a subpopulation of amniotic fluid-derived stem cells (AF-SCs) at the single-cell level by limiting dilution. We found that NANOG- and POU5F1 (also known as OCT4)-expressing cells still existed in the expanded single cell-derived AF-SCs. Aside from the common mesenchymal characteristics, these clonal AF-SCs also exhibit multiple phenotypes of neural-derived cells such as NES, TUBB3, NEFH, NEUNA60, GALC, and GFAP expressions both before and after neural induction. Most importantly, HPLC analysis showed the evidence of dopamine release in the extract of dopaminergic-induced clonal AF-SCs. The results of this study suggest that besides being an easily accessible and expandable source of fetal stem cells, amniotic fluid will provide a promising source of neural progenitor cells that may be used in future cellular therapies for neurodegenerative diseases and nervous system injuries. PMID- 16306423 TI - Placental insufficiency leads to developmental hypertension and mesenteric artery dysfunction in two generations of Sprague-Dawley rat offspring. AB - It is generally accepted that preeclampsia results from reduction in perfusion to the uteroplacental unit leading to maternal hypertension and fetal growth restriction. Placental insufficiency creates an environment of fetal undernutriton, predisposing the fetus to the development of adult disease. In this study, we characterized the development and perpetuation of hypertension in two generations of male and female offspring subjected to an environment of fetal undernutrition via reduced uteroplacental perfusion pressure. Further, we examined vascular responses of resistance arteries in these animals to determine the influence of placental insufficiency on the development and perpetuation of hypertension. Experimental dams underwent a surgical procedure to reduce uteroplacental perfusion pressure, with resulting offspring comprising the first generation (F1). One male and one female from each of the F1 experimental litters served as breeders of the second generation (F2). Weekly systolic blood pressure measurements were obtained from 4 to 24 wk in control, F1, and F2 offspring. Vascular responsiveness to the vasoconstrictors phenylephrine and potassium chloride and the vasorelaxants acetylcholine and sodium nitroprusside was determined in the three offspring groups at 6, 9, and 12 wk of age. Our findings indicate that placental insufficiency during a critical developmental window in late gestation leads to hypertension in juvenile Sprague-Dawley rat offspring and is perpetuated in a second generation of offspring in a gender-specific manner. Further, exposure to placental insufficiency during late gestation leads to developmental alterations characterized by vascular hyperresponsiveness, perpetuated to a second generation of offspring in the absence of persistent environmental stimuli, contributing to hypertension. PMID- 16306424 TI - Expression and glycosylation with polylactosamine of CD44 antigen on macrophages during follicular atresia in pig ovaries. AB - Macrophages are essential in cleaning up apoptotic debris during follicular atresia. However, the key factors of this process are still unclear. In the present study, we evaluated CD44 mRNA, CD44 protein, and CD44 antigen glycosylation on macrophages during follicular atresia in the pig. Atresia was classified into five stages: stage I, healthy follicles; stage II, early atretic follicles having apoptotic granulosa cells with an unclear basement membrane; stage III, progressing atretic follicles having apoptotic granulosa cells completely diffused from the basement membrane; stage IV, late atretic follicles with increasing lysosomal activity; and stage V, disintegrated atretic follicles having collapsed theca cells and strong lysosomal activity. Immunohistological analysis showed that macrophages expressing CD44 invaded the inside of stage III follicles, accompanied by a collapse of basement membrane. Semiquantitative RT PCR showed that only mRNA of the CD44 standard isoform (CD44s) was present in inner cells of follicles, and not any CD44 variant isoform (CD44v) mRNAs. The amount of CD44s mRNA was increased at stage III. Western blot and lectin blot analyses showed that CD44 was markedly expressed at stage III and glycosylated with polylactosamine at the same time. After macrophages invaded atretic follicles at stages III-V, the CD44 expressed on macrophages was glycosylated with polylactosamine. The lysosomal activity began to increase at stage IV, and reached the highest level at stage V. Increased CD44s protein and posttranslational modification of CD44 with polylactosamine on macrophages from stage III could be involved in the cleaning up apoptotic granulosa cells. PMID- 16306425 TI - Influence of TRP53 status on FAS membrane localization, CFLAR (c-FLIP) ubiquitinylation, and sensitivity of GC-2spd (ts) cells to undergo FAS-mediated apoptosis. AB - Previously we reported that testicular germ cells undergo FAS-mediated apoptosis after exposure of mice to the Sertoli cell toxicant mono-(2-ethylhexyl) phthalate (MEHP) and that this process is partially dependent on the TRP53 protein (p53). Recent reports have suggested that TRP53 may influence the ubiquitinylation and consequent proteosomal degradation of a negative regulator of FAS, CFLAR (L) (c FLIP [L]), in human colon cancer cells. To further characterize the relationship between CFLAR and TRP53, we used the transformed germ cell line GC-2spd (ts), which harbors a temperature-sensitive Trp53 mutation that allows for TRP53 activation at 32 degrees C. We report here that GC-2 cells expressed a 10-fold increase in basal cell membrane FAS levels and an increased sensitivity to FAS agonistic antibody (JO2)-triggered apoptosis only when they were maintained at the permissive TRP53 temperature. After JO2 exposure, CFLAR (L) protein levels were enhanced only at the nonpermissive TRP53 temperature (37 degrees C) while real-time PCR results indicated an absence of Cflar (L) mRNA changes in GC-2 cells regardless of the temperature. Furthermore, transfection of GC-2 cells at 37 degrees C with siRNA against Cflar resulted in reduction of CFLAR (L) protein levels and increased sensitivity to JO2-mediated apoptosis. The CFLAR (L) protein was also more strongly ubiquitinylated in response to JO2 treatment at the permissive TRP53 temperature. Taken together, these data suggest that the TRP53 protein influences the sensitivity of GC-2 cells to undergo FAS-mediated apoptosis by modulating the expression of FAS on their cell membranes and subsequently influencing the degradation of the antiapoptotic protein CFLAR (L). PMID- 16306426 TI - The apolipoprotein B/AI ratio and the metabolic syndrome independently predict risk for myocardial infarction in middle-aged men. AB - BACKGROUND: Both the metabolic syndrome and an increased apolipoprotein B/AI (apoB/AI) ratio are powerful risk factors for cardiovascular events. We hypothesized that the apoB/AI ratio well-characterizes the dyslipidemia associated with insulin resistance and the metabolic syndrome and investigated those relations and if the apoB/AI ratio and the metabolic syndrome independently predicted subsequent myocardial infarction (MI). METHODS AND RESULTS: A community based sample of 1826 men aged 50 was investigated at baseline and again at age 70. ApoB/AI ratio and the metabolic syndrome (National Cholesterol Education Program definition) were evaluated, and the incidence of fatal and nonfatal MI was followed for a median of 26.8 years from the age 50 baseline. ApoB/AI ratio was significantly higher in men with versus without the metabolic syndrome (P<0.0001), and increased with the number of components defining the syndrome (P<0.0001). ApoB/AI ratio was inversely related to euglycemic insulin clamp glucose disposal rate at age 70 (r=-0.34, P<0.0001). During follow-up from age 50, 462 subjects developed an MI. An apoB/AI ratio > or =0.9 (hazard ratio [HR], 1.48; 95% confidence interval [CI], 1.15 to 1.91) and presence of the metabolic syndrome (HR, 1.69; 95% CI, 1.30 to 2.21) at baseline were independent predictors for MI, adjusting for low-density lipoprotein cholesterol and smoking. CONCLUSIONS: The apoB/AI ratio was related to the metabolic syndrome, as well as to a direct measurement of insulin resistance. Despite this, the apoB/AI ratio and the metabolic syndrome were both independent long-term predictors of MI in a community-based sample of middle-aged men. PMID- 16306427 TI - Importance of junctional adhesion molecule-A for neointimal lesion formation and infiltration in atherosclerosis-prone mice. AB - OBJECTIVE: Although junctional adhesion molecule-A (JAM-A) has recently been implicated in leukocyte recruitment on early atherosclerotic endothelium and after reperfusion injury, its role in neointima formation after arterial injury remains to be elucidated. METHODS AND RESULTS: Here we show that the genetic deletion of JAM-A in apolipoprotein E-deficient (apoE(-/-)) mice significantly reduced neointimal hyperplasia after wire injury of carotid arteries without altering medial area. This was associated with a significant decrease in neointimal macrophage content, whereas the relative content of smooth muscle cells and endothelial recovery was unaltered in JAM-A(-/-)apoE(-/-) compared with JAM-A(+/+)apoE(-/-) lesions. In carotid arteries perfused ex vivo, deficiency in JAM-A significantly impaired the recruitment of monocytes 1 week, but not 1 day, after injury. These effects were paralleled by an attenuation of monocyte arrest and transmigration on activated JAM-A(-/-)apoE(-/-) versus JAM-A(+/+)apoE(-/-) endothelial cells under flow conditions in vitro. A mechanism underlying reduced recruitment was implied by findings that the luminal expression of the arrest chemokine RANTES in injured arteries and its endothelial deposition by activated platelets in vitro were diminished by JAM-A deficiency. CONCLUSIONS: Our data provide the first evidence to our knowledge for a crucial role of JAM-A in accelerated lesion formation and monocyte infiltration in atherosclerosis-prone mice. PMID- 16306428 TI - Elevated plasma membrane cholesterol content alters macrophage signaling and function. AB - OBJECTIVE: During atherogenesis, macrophages migrate into the subendothelial space where they ingest deposited lipoproteins, accumulate lipids, and transform into foam cells. It is unclear why these macrophages do not remove their lipid loads from the region. This study was aimed at testing the hypothesis that macrophage behavior is altered when membrane cholesterol levels are elevated, as might be the case for cells in contact with lipoproteins within atherosclerotic lesions. METHODS AND RESULTS: We examined the effects of elevating membrane cholesterol on macrophage behavior. J774 macrophages were treated with either acetylated low-density lipoprotein (ac-LDL) and ACAT inhibitor or cholesterol chelated methyl-beta-cyclodextrin (chol-MbetaCD) to increase membrane cholesterol levels. Our results show that elevating the membrane cholesterol of J774 macrophages induced dramatic ruffling, stimulated cell spreading, and affected F actin organization. Cellular adhesion was required for these effects, and Rac mediated signaling pathways were involved. Additionally, 3-dimensional transwell chemotaxis assays showed that migration of J774 macrophages was significantly inhibited when membrane cholesterol levels were raised. CONCLUSIONS: These findings indicate that increased membrane cholesterol causes dramatic effects on macrophage cellular functions related to the actin cytoskeleton. They should provide new insights into the early steps of atherogenesis. PMID- 16306429 TI - Vascular function and mild renal impairment in stable coronary artery disease. AB - OBJECTIVE: In patients with coronary artery disease, the concomitant presence of renal function impairment is associated with decreased survival. We aimed to assess whether in coronary artery diseased patients renal function impairment is associated with systemic vascular function, functional parameters of the renin angiotensin system, or inflammation as potential mediators for cardiovascular risk. METHODS AND RESULTS: We studied 125 patients, 87% male, with a mean age of 62.2+/-8.2 years; 72% had 3-vessel disease, and mean renal function was 74+/-13 mL/min per 1.73 m2. Internal thoracic artery rings were sampled during coronary bypass surgery and used for in vitro vascular measurements. We could not establish an association between endothelium-dependent vasorelaxation (response to methacholine) and renal function. In addition, vascular response to potassium chloride, phenylephrine, and angiotensin II were not associated with renal function. Finally, serum angiotensin-converting enzyme (ACE) activity, usage of ACE inhibitors, C-reactive protein, and soluble intercellular adhesion molecule 1 were not related to renal function. CONCLUSIONS: In coronary artery disease patients, mild renal function impairment is not associated with systemic vasomotor responsiveness, inflammation, or functional systemic parameters of the renin-angiotensin system. The relation between systemic endothelial dysfunction and mild renal insufficiency might be more complicated than previously thought. PMID- 16306430 TI - Lesional overexpression of matrix metalloproteinase-9 promotes intraplaque hemorrhage in advanced lesions but not at earlier stages of atherogenesis. AB - BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is involved in atherosclerosis and elevated MMP-9 activity has been found in unstable plaques, suggesting a crucial role in plaque rupture. This study aims to assess the effect of MMP-9 on plaque stability in apolipoprotein E-deficient mice at different stages of plaque progression. METHODS AND RESULTS: Atherosclerotic lesions were elicited in carotid arteries by perivascular collar placement. MMP-9 overexpression in intermediate or advanced plaques was effected by intraluminal incubation with an adenovirus (Ad.MMP-9). A subset was coincubated with Ad.TIMP-1. Mock virus served as a control. Plaques were analyzed histologically. In intermediate lesions, MMP 9 overexpression induced outward remodeling, as shown by a 30% increase in media size (p=0.03). In both intermediate and advanced lesions, prevalence of vulnerable plaque morphology tended to be increased. Half of MMP-9-treated lesions displayed intraplaque hemorrhage, whereas in controls and the Ad.MMP 9/Ad.TIMP-1 group this was 8% and 16%, respectively (p=0.007). Colocalization with neovessels may point to neo-angiogenesis as a source for intraplaque hemorrhage. CONCLUSIONS: These data show a differential effect of MMP-9 at various stages of plaque progression and suggest that lesion-targeted MMP-9 inhibition might be a valuable therapeutic modality in stabilizing advanced plaques, but not at earlier stages of lesion progression. PMID- 16306431 TI - Isolation of "side population" progenitor cells from healthy arteries of adult mice. AB - OBJECTIVE: Circulating progenitors and stem cells have been reported to contribute to angiogenesis and arterial repair after injury. In the present study, we investigated whether the arterial wall could host permanently residing progenitor cells under physiological context. METHODS AND RESULTS: Using the Hoechst-based flow cytometry method, we identified and isolated progenitor cells termed side population (SP) cells at a prevalence of 6.0+/-0.8% in the tunica media of adult mice aortas. Arterial SP cells expressed the ATP-binding cassette transporter subfamily G member 2, frequently present on SP cell surface, and displayed a Sca-1+ c-kit(-/low) Lin- CD34(-/low) profile. They did not form myeloid or lymphoid hematopoietic colonies after plating in methylcellulose-based medium. Importantly, cultured SP cells were able to acquire the phenotype of endothelial cells (CD31, VE-cadherin, and von Willebrand factor expression) or of smooth muscle cells (alpha-smooth muscle actin, calponin, and smooth muscle myosin heavy chain expression), in presence of either vascular endothelial growth factor or transforming growth factor (TGF)-beta1/PDGF-BB, respectively. Moreover, they generated vascular-like branching structures, composed of both VE-cadherin+ cells and alpha-smooth muscle actin+ cells on Matrigel. CONCLUSIONS: In this study, we provide the first evidence to our knowledge that in the adult mice, the normal arterial wall harbors SP cells with vascular progenitor properties. PMID- 16306432 TI - Cell-based ex vivo delivery of angiogenic growth factors for cardiac repair. PMID- 16306433 TI - Early anticoagulant effect of atorvastatin. PMID- 16306434 TI - Mechanisms of statin-induced myopathy: a role for the ubiquitin-proteasome pathway? PMID- 16306435 TI - Apolipoprotein AV: low concentration, high impact. PMID- 16306436 TI - Killing two birds with one stone: targeting chemokine receptors in atherosclerosis and HIV infection. PMID- 16306437 TI - Dissociation between the short-term effects of nocturnal hemodialysis on endothelium dependent vasodilation and plasma ADMA. PMID- 16306438 TI - Associations between liver histology and carotid intima-media thickness in patients with nonalcoholic fatty liver disease. PMID- 16306439 TI - Heparin cofactor II levels do not predict the development of coronary heart disease: the Atherosclerosis Risk in Communities (ARIC) study. PMID- 16306442 TI - Diabetic microangiopathy in ischemic limb is a disease of disturbance of the platelet-derived growth factor-BB/protein kinase C axis but not of impaired expression of angiogenic factors. AB - Diabetic foot is caused by microangiopathy and is suggested to be a result of impaired angiogenesis. Using a severe hindlimb ischemia model of streptozotocin induced diabetic mice (STZ-DM), we show that diabetic foot is a disease solely of the disturbance of platelet-derived growth factor B-chain homodimer (PDGF-BB) expression but not responses of angiogenic factors. STZ-DM mice frequently lost their hindlimbs after induced ischemia, whereas non-DM mice did not. Screening of angiogenesis-related factors revealed that only the expression of PDGF-BB was impaired in the STZ-DM mice on baseline, as well as over a time course after limb ischemia. Supplementation of the PDGF-B gene resulted in the prevention of autoamputation, and, furthermore, a protein kinase C (PKC) inhibitor restored the PDGF-BB expression and also resulted in complete rescue of the limbs of the STZ DM mice. Inhibition of overproduction of advanced-glycation end product resulted in dephosphorylation of PKC-alpha and restored expression of PDGF-BB irrespective of blood sugar and HbA1c, indicating that advanced-glycation end product is an essential regulator for PKC/PDGF-BB in diabetic state. These findings are clear evidence indicating that diabetic vascular complications are caused by impairment of the PKC/PDGF-B axis, but not by the impaired expression of angiogenic factors, and possibly imply the molecular target of diabetic foot. PMID- 16306443 TI - Antihypertensive effects of the putative T-type calcium channel antagonist mibefradil are mediated by the L-type calcium channel Cav1.2. AB - The role of T-type Ca2+ channels for cardiovascular physiology, in particular blood pressure regulation, is controversial. Selective blockade of T-type Ca2+ channels in resistance arteries has been proposed to explain the effect of the antihypertensive drug mibefradil. In the present study, we used a third generation, time- and tissue-specific conditional knockout model of the L-type Ca2+ channel Cav1.2 (Cav1.2SMAKO mice) to genetically dissect the effects of mibefradil on T- and L-type Ca2+ channels. Myogenic tone and phenylephrine induced contraction in hindlimb perfusion experiments were sensitive to mibefradil in control mice, whereas the drug showed no effect in Cav1.2-deficient animals. Mean arterial blood pressure in awake, freely moving control mice was reduced by 38+/-2.5 mm Hg at a dose of 1.25 mg/kg bodyweight mibefradil, but not changed in Cav1.2SMAKO mice. These results demonstrate that the effect of the putative T-type Ca2+ channel-selective blocker mibefradil on blood pressure and small vessel myogenic tone is mediated by the Cav1.2 L-type Ca2+ channel. PMID- 16306444 TI - Hypoxia inducible factor-1 activation by prolyl 4-hydroxylase-2 gene silencing attenuates myocardial ischemia reperfusion injury. AB - Hypoxia inducible factor-1 (HIF-1) regulates changes in transcription of key genes such as inducible NO synthase (iNOS) in hypoxic/ischemic environments. In normoxia, HIF-1 activation is controlled by HIF-1alpha-prolyl 4-hydroxylases, which target HIF-1alpha for ubiquitination and proteasomal degradation. We hypothesized that normoxic HIF-1 preservation could attenuate cardiac ischemia/reperfusion injury via a preconditioning effect. HIF-1 preservation was achieved by using small interfering RNA (siRNA) to silence murine HIF-1alpha prolyl-4 hydroxylase-2 (PHD2). PHD2 siRNA reduced PHD2 mRNA expression 89+/-1.5% (P<0.001) in a time- and concentration-dependent manner in normoxic murine microvascular endothelial cells (EC). PHD2 silencing in normoxic EC stabilized HIF-1alpha protein levels while significantly increasing HIF-1 transcriptional activity and iNOS mRNA expression. Wild-type mice infused with PHD2 siRNA (1.5 microg/g body weight) showed a 61+/-2.4% (P<0.05) reduction in cardiac PHD2 mRNA within 24 hours. In addition HIF-1alpha protein levels and HIF-1-dependent iNOS mRNA levels were increased. PHD2 siRNA-transfected hearts from wild-type mice (n=6) subjected to 30 minutes ischemia followed by 60 minutes reperfusion exhibited reduced infarct size when compared with saline-treated controls (9.7+/ 1.9% versus 31.6+/-1.8%, respectively, P<0.0001, n=6) and to control mice transfected with a nontargeting siRNA control (28.4+/-3.0%, P<0.0001, n=6). Hearts from iNOS knockout mice receiving PHD2 siRNA by identical injection protocol (n=6) exhibited infarct size indistinguishable from saline controls (28.7+/-1.3%). These results show that in vitro and in vivo, PHD2 silencing using a siRNA strategy produces transcriptionally active HIF-1. Normoxic activation of HIF-1 in hearts following in vivo PHD2 siRNA administration attenuates reperfusion injury via an iNOS-dependent pathway. PMID- 16306445 TI - Microtubules modulate the stiffness of cardiomyocytes against shear stress. AB - Although microtubules are involved in various pathological conditions of the heart including hypertrophy and congestive heart failure, the mechanical role of microtubules in cardiomyocytes under such conditions is not well understood. In the present study, we measured multiple aspects of the mechanical properties of single cardiomyocytes, including tensile stiffness, transverse (indentation) stiffness, and shear stiffness in both transverse and longitudinal planes using carbon fiber-based systems and compared these parameters under control, microtubule depolymerized (colchicine treated), and microtubule hyperpolymerized (paclitaxel treated) conditions. From all of these measurements, we found that only the stiffness against shear in the longitudinal plane was modulated by the microtubule cytoskeleton. A simulation model of the myocyte in which microtubules serve as compression-resistant elements successfully reproduced the experimental results. In the complex strain field that living myocytes experience in the body, observed changes in shear stiffness may have a significant influence on the diastolic property of the diseased heart. PMID- 16306446 TI - Cardiomyocyte cell cycle activation ameliorates fibrosis in the atrium. AB - MHC-TGFcys33ser transgenic mice have elevated levels of active transforming growth factor (TGF)-beta1 in the myocardium. Previous studies have shown that these animals develop atrial, but not ventricular, fibrosis. Here we show that atrial fibrosis was accompanied with cardiomyocyte apoptosis. Although similar levels of cardiomyocyte apoptosis were present in the right and left atria of MHC TGFcys33ser hearts, the extent of fibrosis was more pronounced in the right atrium. Thus, additional factors influence the degree of atrial fibrosis in this model. Tritiated thymidine incorporation studies revealed cardiomyocyte cell cycle activity in left atrial cardiomyocytes, but not in right atrial cardiomyocytes. These observations suggested that cardiomyocyte cell cycle activation ameliorated the severity of atrial fibrosis. To directly test this hypothesis, MHC-TGFcys33ser mice were crossed with MHC-cycD2 mice (which have constitutive cardiomyocyte cell cycle activity in the right atrium). Mice inheriting both transgenes exhibited right atrial cardiomyocyte cell cycle activity and a concomitant reduction in the severity of right atrial fibrosis, despite the presence of a similar level of cardiomyocyte apoptosis as was observed in mice inheriting the MHC-TGFcys33ser transgene alone. These data support the notion that cardiomyocyte cell cycle induction can antagonize fibrosis in the myocardium. PMID- 16306447 TI - c-Jun N-terminal kinases mediate reactivation of Akt and cardiomyocyte survival after hypoxic injury in vitro and in vivo. AB - Akt is a central regulator of cardiomyocyte survival after ischemic injury in vitro and in vivo, but the mechanisms regulating Akt activity in the postischemic cardiomyocyte are not known. Furthermore, although much is known about the detrimental role that the c-Jun N-terminal kinases (JNKs) play in promoting death of cells exposed to various stresses, little is known of the molecular mechanisms by which JNK activation can be protective. We report that JNKs are necessary for the reactivation of Akt after ischemic injury. We identified Thr450 of Akt as a residue that is phosphorylated by JNKs, and the phosphorylation status of Thr450 regulates reactivation of Akt after hypoxia, apparently by priming Akt for subsequent phosphorylation by 3-phosphoinositide-dependent protein kinase. The reduction in Akt activity that is induced by JNK inhibition may have significant biological consequences, as we find that JNKs, acting via Akt, are critical determinants of survival in posthypoxic cardiomyocytes in culture. Furthermore, in contrast to selective p38-mitogen-activated protein kinase inhibition, which was cardioprotective in vivo, concurrent inhibition of both JNKs and p38-mitogen activated protein kinases increased ischemia/reperfusion injury in the heart of the intact rat. These studies demonstrate that reactivation of Akt after resolution of hypoxia and ischemia is regulated by JNKs and suggest that this is likely a central mechanism of the myocyte protective effect of JNKs. PMID- 16306448 TI - Intracellular calcium handling dysfunction and arrhythmogenesis: a new challenge for the electrophysiologist. PMID- 16306449 TI - Cardiac progenitor cells: the revolution continues. PMID- 16306450 TI - The translation of transcription. PMID- 16306451 TI - An apolipoprotein A-I mimetic works best in the presence of apolipoprotein A-I. PMID- 16306452 TI - NO to small mothers against decapentaplegic (Smad). PMID- 16306453 TI - Endothelial extracellular matrix: biosynthesis, remodeling, and functions during vascular morphogenesis and neovessel stabilization. AB - The extracellular matrix (ECM) is critical for all aspects of vascular biology. In concert with supporting cells, endothelial cells (ECs) assemble a laminin-rich basement membrane matrix that provides structural and organizational stability. During the onset of angiogenesis, this basement membrane matrix is degraded by proteinases, among which membrane-type matrix metalloproteinases (MT-MMPs) are particularly significant. As angiogenesis proceeds, ECM serves essential functions in supporting key signaling events involved in regulating EC migration, invasion, proliferation, and survival. Moreover, the provisional ECM serves as a pliable scaffold wherein mechanical guidance forces are established among distal ECs, thereby providing organizational cues in the absence of cell-cell contact. Finally, through specific integrin-dependent signal transduction pathways, ECM controls the EC cytoskeleton to orchestrate the complex process of vascular morphogenesis by which proliferating ECs organize into multicellular tubes with functional lumens. Thus, the composition of ECM and therefore the regulation of ECM degradation and remodeling serves pivotally in the control of lumen and tube formation and, finally, neovessel stability and maturation. PMID- 16306455 TI - Stroke attributable to a calcific embolus from the brachiocephalic trunk. AB - BACKGROUND AND PURPOSE: Calcific brain embolization is a rare event that is usually secondary to cardiac valve calcification. We present a case of stroke caused by embolization of calcific material from the brachiocephalic trunk, probably induced by radiotherapy. SUMMARY OF CASE: A 56-year-old right-handed female developed left-sided hemiparesis, hemihypesthesia, and sensory inattention. She had a history of right breast carcinoma that was excised 8 years previously followed by radiotherapy. She had no other history of note. Computed tomography of the head and magnetic resonance imaging confirmed a calcific embolus in right middle cerebral artery and an acute infarction in the corresponding territory. Plain chest radiography, carotid ultrasonography, transthoracic and transoesophageal echocardiography failed to demonstrate the source of calcific embolism. Computed tomography of the thorax revealed heavy calcification of the brachiocephalic trunk and the origin of the right common carotid artery. CONCLUSIONS: Undertaking a vigilant systematic search for the source in cases of calcific embolization is necessary. The aorta and its main branches are possible, yet unusual, sources of calcific emboli that merit investigation. PMID- 16306456 TI - Prophylaxis of thrombotic and embolic events in acute ischemic stroke with the low-molecular-weight heparin certoparin: results of the PROTECT Trial. AB - BACKGROUND AND PURPOSE: Patients with stroke are at substantial risk of thromboembolic complications and therefore require antithrombotic prophylaxis. To show the noninferiority of the low-molecular-weight heparin certoparin to unfractionated heparin (UFH) for the prevention of thromboembolic complications, we performed a randomized, double-blind, active-controlled multicenter trial in patients with acute ischemic stroke. METHODS: Overall, 545 patients were randomized within 24 hours of stroke onset to treatment with certoparin (3000 U anti-Xa OD; n=272) or UFH (5000 U TID; n=273) for 12 to 16 days. Patients with paresis of a leg and an National Institutes of Health Stroke Scale score of 4 to 30 points were included. The primary end point was a composite outcome of proximal deep vein thrombosis, pulmonary embolism, or death related to venous thromboembolism during treatment. Computed tomography was performed at trial entry, after 7 days, and when clinical deterioration occurred. RESULTS: The per protocol analysis revealed 17 (7.0%) primary events in the certoparin group compared with 24 (9.7%) in the UFH group, thereby demonstrating noninferiority (P=0.0011), confirmed by intention-to-treat analysis (6.6% versus 8.8%; P=0.008). Major bleeding occurred during treatment in 3 patients allocated to certoparin (1.1%) and 5 patients allocated to UFH (1.8%). CONCLUSIONS: Certoparin (3000 U anti-Xa OD) is at least as effective and safe as UFH (TID) for the prevention of thromboembolic complications in patients with acute ischemic stroke. PMID- 16306457 TI - Ataxic hemiparesis from strategic frontal white matter infarction with crossed cerebellar diaschisis. AB - BACKGROUND: Ataxic hemiparesis is a classic lacunar syndrome that most often localizes to the pons. RESULTS: We report 3 patients who presented with left sided ataxic hemiparesis and were found on imaging to have small right frontal subcortical white matter infarcts in similar locations by diffusion-weighted MRI. [99mTc]hexamethylpropylenamine oxime single-photon emission computed tomography (SPECT) scans in all 3 patients showed decreased metabolism in the contralateral cerebellar hemisphere, indicative of crossed cerebellar diaschisis (CCD). CONCLUSIONS: CCD is under-recognized in ataxic hemiparesis and may have implications for functional recovery after this type of ischemic stroke. PMID- 16306458 TI - Improved interpretation of stroke trial results using empirical Barthel item weights. AB - BACKGROUND AND PURPOSE: Attempts have been made to provide guidelines for interpreting Barthel scores. We used a Rasch analysis to improve the measurement properties and clinical interpretability of the Barthel index score. METHODS: A specific extension of Rasch model was used to identify items that preclude the summation of items and to improve the item rating scale by examining the scores on the Barthel of 559 stroke patients scored 3 weeks (n=89) and 6 months (n=470) after stroke. The clinical interpretation of the revised Rasch modeled Barthel was illustrated by re-examining the results of a previously published trial on the effectiveness of leg and arm training after stroke. RESULTS: Most rating scales could be improved by collapsing nondiscriminating rating categories. Two items showed misfit: Bladder and Bowel. The remaining Barthel showed an excellent fit to the extended Rasch model (R1c Goodness-of-Fit P=0.35). Both items and patients could be placed on a common logit unit scale, allowing a clearer interpretation of the trial effect. Using the modeled activities of daily living difficulty/ability scale, we could express the differences between treatment arms in modeled probabilities of a positive score to each Barthel item for the treatment arms not conveyed by the original ordinal Barthel sum scores. CONCLUSIONS: We improved the psychometric properties and clinical interpretation of the Barthel index. PMID- 16306459 TI - Hyperglycemia, insulin, and acute ischemic stroke: a mechanistic justification for a trial of insulin infusion therapy. AB - BACKGROUND AND PURPOSE: Hyperglycemia is associated with increased mortality and morbidity in acute ischemic stroke. SUMMARY OF REVIEW: Hyperglycemia induces a pro-oxidative and proinflammatory state that can cause direct neuronal toxicity. Hyperglycemia-mediated increase in matrix metalloproteinase-9 can cause neuronal damage by an increase in cerebral edema. Moreover, hyperglycemia may be responsible for a procoagulant state that can further compromise blood supply to the penumbral areas in acute ischemic stroke. Insulin infusion has an effect that is opposite to that of hyperglycemia. It not only lowers blood glucose levels but also exerts an antioxidant and anti-inflammatory effect. Insulin also improves NO production and results in improved blood circulation to the ischemic areas. This article focuses on the potential mechanisms underlying the injurious effects of glucose and the beneficial effects of insulin. CONCLUSIONS: In the absence of other potential beneficial therapies, there is an urgency to institute trials with insulin infusion in acute ischemic stroke. PMID- 16306460 TI - Is there a causal relationship between the amount of alcohol consumption and stroke risk? PMID- 16306461 TI - Treating warfarin-related intracerebral hemorrhage: is fresh frozen plasma enough? PMID- 16306462 TI - Activity in the peri-infarct rim in relation to recovery from stroke. AB - BACKGROUND AND PURPOSE: In the rim of tissue surrounding a cortical infarct, animal studies have described an increase in a number of growth-related processes that likely contribute to behavioral recovery. The current study hypothesized that in patients with good outcome after stroke, brain activation in peri-infarct tissue would be greater than normal. METHODS: In 15 patients with good recovery chronically after ischemic cortical stroke, activation within peri-infarct brain tissue was directly compared with activation within the same brain tissue of 13 control subjects. RESULTS: Although most patients did show activation within peri infarct tissues, their activation compared with controls was reduced rather than increased. Evaluation of the T2*-weighted images underlying functional MRI mapping disclosed a significant gradient of increased T2* signal in peri-infarct tissues, likely attributable to tissue changes such as gliosis. CONCLUSIONS: Among well-recovered stroke patients, cortical activation is present in the area surrounding a cortical infarct but is smaller than normal. A baseline derangement of the T2*-weighted signal underlying functional MRI (fMRI) is also present in this area, which might influence interpretation of fMRI findings. The relationship between increased tissue T2* signal and fMRI activation is not known and requires further study. PMID- 16306463 TI - Retinal microvascular abnormalities and MRI-defined subclinical cerebral infarction: the Atherosclerosis Risk in Communities Study. AB - BACKGROUND AND PURPOSE: Retinal microvascular abnormalities reflect cumulative small vessel damage from elevated blood pressure and may reflect subclinical cerebral microvascular changes. We examined their associations with MRI-defined cerebral infarcts. METHODS: Population-based, cross-sectional study of 1684 persons 55 to 74 years of age without a history of clinical stroke, sampled from 2 US southeastern communities. Retinal photographs were obtained and graded for presence of retinal microvascular abnormalities, including arteriovenous nicking, focal arteriolar narrowing, retinal hemorrhages, soft exudates and microaneurysms. Photographs were also digitized, and retinal vessel diameters were measured and summarized as the arteriole-to-venule ratio (AVR). Cerebral MRI scans were graded for presence of cerebral infarct, defined as a lesion > or =3 mm diameter in a vascular distribution with typical imaging characteristics. RESULTS: There were a total of 183 MRI cerebral infarcts. After adjustment for age, gender, race, 6-year mean arterial blood pressure, diabetes, and other stroke risk factors, cerebral infarcts were associated with retinal microvascular abnormalities, with odds ratios 1.90 (95% CI, 1.25 to 2.88) for arteriovenous nicking, 1.89 (95% CI, 1.22 to 2.92) for focal arteriolar narrowing, 2.95 (95% CI, 1.30 to 6.71) for blot hemorrhages, 2.08 (95% CI, 0.69, 6.31) for soft exudates, 3.17 (95% CI, 1.05 to 9.64) for microaneurysms, and 1.74 (95% CI, 0.95 to 3.21) for smallest compared with largest AVR. In stratified analyses, these associations were only present in persons with hypertension. CONCLUSIONS: Retinal microvascular abnormalities are associated with MRI-defined subclinical cerebral infarcts independent of stroke risk factors. These data suggest that retinal photography may be useful for studying subclinical cerebrovascular disease in population-based studies. PMID- 16306464 TI - Moderate alcohol consumption reduces risk of ischemic stroke: the Northern Manhattan Study. AB - BACKGROUND AND PURPOSE: Moderate alcohol consumption is protective against coronary disease, but its relationship to ischemic stroke (IS) is controversial. METHODS: Stroke-free participants > or =40 years of age identified by random digit dialing were enrolled in a prospective cohort study between 1993 and 2001. Alcohol consumption was assessed through in-person interview and categorized as none in the past year, > or =1 drink in past month to < or =2 per day (moderate drinkers), and >2 drinks daily. Lifetime drinking was also assessed. Cox proportional hazard regression modeling was used to assess hazard ratios and their 95% CIs for the association of drinking with risk of stroke and vascular events. RESULTS: Mean age among participants (n=3176) was 69.1+/-10.3 years; 62.8% were women, 20.8% were non-Hispanic white, 24.5% non-Hispanic black, and 52.4% were Hispanic. No alcohol in the previous year was present in 62.3%, and 32.5% drank moderately. After adjusting for other risk factors compared with those who did not drink in the past year, moderate drinkers had a reduced risk of IS (0.67; 95% CI, 0.46 to 0.99) and IS, myocardial infarction, or vascular death (0.74; 95% CI, 0.59 to 0.94). Results were similar when never-drinkers were used as referent group. Reduction in risk was seen for nonatherosclerotic IS subtypes, and results stratified by age, sex, and race-ethnicity were similar. CONCLUSIONS: Moderate alcohol consumption is associated with decreased risk of IS in a multiethnic population. This effect is independent of other risk factors and holds for nonatherosclerotic stroke subtypes. PMID- 16306465 TI - Timing of fresh frozen plasma administration and rapid correction of coagulopathy in warfarin-related intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Anticoagulation-related intracerebral hemorrhage (ICH) is often fatal, and rapid reversal of anticoagulation is the most appealing strategy currently available for treatment. We sought to determine whether particular emergency department (ED) interventions are effective in reversing coagulopathy and improving outcome. METHODS: Consecutive patients with warfarin-related ICH presenting to an urban tertiary care hospital from 1998 to 2004 were prospectively captured in a database. ED records were retrospectively reviewed for dose and timing of fresh-frozen plasma (FFP) and vitamin K, as well as serial coagulation measures. After excluding patients with incomplete ED records, do-not resuscitate orders established in the ED, initial international normalized ratio (INR) < or =1.4, and for whom no repeat INR was performed, 69 patients were available for analysis. The primary outcome was a documented INR < or =1.4 within 24 hours of ED presentation. RESULTS: Patients whose INR was successfully reversed within 24 hours had a shorter median time from diagnosis to first dose of FFP (90 minutes versus 210 minutes; P=0.02). In multivariable analysis, shorter time to vitamin K, as well as FFP, predicted INR correction. Every 30 minutes of delay in the first dose of FFP was associated with a 20% decreased odds of INR reversal within 24 hours (odds ratio, 0.8; 95% CI, 0.63 to 0.99). Dosing of FFP and vitamin K had no effect. No ED intervention was associated with improved clinical outcome. CONCLUSIONS: Time to treatment is the most important determinant of 24-hour anticoagulation reversal. Although additional study is required to determine the clinical benefit of rapid reversal of anticoagulation, minimizing delays in FFP administration is a prudent first step in emergency management of warfarin-related ICH. PMID- 16306466 TI - Antiplatelet drugs in the secondary prevention after stroke: differential efficacy in large versus small vessel disease? A subgroup analysis from ESPS-2. AB - BACKGROUND AND PURPOSE: Arterial disease resulting in cerebral ischemia can be classified into large vessel disease (LVD) and small vessel disease (SVD). We assessed whether antiplatelet drugs were more efficacious in large than in small vessel cerebrovascular disease. METHODS: Individual patient data of the second European Stroke Prevention Study (n=6602), in which patients with a previous transient ischemic attack or ischemic stroke were randomized to aspirin, dipyridamole, their combination, or placebo, were reanalyzed. Type of vessel disease was classified according to clinical symptoms or physical examination. Presence of a lacunar syndrome was considered typical for SVD and evidence of cortical dysfunction for LVD. Vascular events (nonfatal stroke, nonfatal myocardial infarction, nonfatal other vascular event, or vascular death) were taken as outcome. Cox regression analyses were performed. RESULTS: A total of 419 first vascular events occurred in 2600 patients with SVD and 367 in 1816 patients with LVD (mean follow-up 1.7 years). For aspirin versus placebo, the hazard ratio (HR) was 0.86 (95% CI, 0.66 to 1.11) in patients with SVD and 0.80 (95% CI, 0.61 to 1.06) in those with LVD (Pinteraction=0.74). For dipyridamole versus placebo, the HR was 0.86 (95% CI, 0.67 to 1.12) in patients with SVD and 0.90 (95% CI, 0.68 to 1.19) in patients with LVD (Pinteraction=0.84). Similar observations were made for the outcome stroke only. CONCLUSIONS: Our findings do not concur with the hypothesis that aspirin, dipyridamole, or the combination may be especially effective in preventing vascular events in patients with previous cerebral ischemia that was caused by LVD compared with SVD. PMID- 16306467 TI - Subtype hypertension and risk of stroke in middle-aged and older Chinese: a 10 year follow-up study. AB - BACKGROUND AND PURPOSE: Hypertension is the most important indicator of stroke. We aim to compare the long-term effects of the subtypes of hypertension on the risk of stroke in a Chinese cohort. METHODS: A total of 26,587 subjects > or =35 years of age and free of stroke were recruited in 5 cities in 1987. The subtypes of hypertension were defined as isolated systolic hypertension (ISH), isolated diastolic hypertension (IDH), systolic and diastolic hypertension (SDH), as well as managed hypertension (MHT), according to the criteria of systolic blood pressure > or =140 or diastolic blood pressure >90 mm Hg or under antihypertensive treatment. The relative risks of stroke with the subtypes of hypertension, compared with normotensives, were estimated using the Cox model after adjustments for age, sex, and other confounders. RESULTS: The prevalence of hypertension was: ISH 7.1%, SDH 18.4%, IDH 6.7%, and MHT 3.9%. During a total of 233 437 person years of follow-up, 1107 subjects developed stroke (614 ischemic and 451 hemorrhagic events and 42 unclassified). SDH patients were at the highest risk of stroke among all the hypertensives. The hazard ratio and 95% CI was 2.96 (2.49 to 3.52) for all stroke, 4.05 (3.10 to 5.30) for hemorrhagic, and 2.33 (1.84 to 2.95) for ischemic stroke. Although the incidence of stroke was higher in the older population, the effect of hypertension, especially SDH, on hemorrhagic stroke is stronger in the middle-aged population. CONCLUSIONS: ISH and IDH are similarly prevalent in the population; both are independent predictors of stroke. Patients with SDH are at the highest risk of stroke and should be treated more aggressively. PMID- 16306468 TI - C-reactive protein and risk of first-ever ischemic and hemorrhagic stroke in a general Japanese population: the Hisayama Study. AB - BACKGROUND AND PURPOSE: The role of high-sensitivity C-reactive protein (hsCRP) in the development of stroke is not clearly understood. We investigated the relationship between serum hsCRP levels and stroke occurrence in a general Japanese population. METHODS: We followed 2692 subjects > or =40 years of age for 12 years. The relative risks and 95% CIs for ischemic and hemorrhagic stroke occurrence were calculated according to the hsCRP quintiles. RESULTS: During the follow-up, 129 first-ever ischemic and 59 hemorrhagic strokes occurred. In men, the age-adjusted incidence of ischemic stroke significantly increased with elevated serum hsCRP levels; the difference between the first and fifth quintiles was statistically significant (1.4 versus 6.6 per 1000 person-years; P=0.02). This association remained significant even after adjustment for other confounding factors, such as age, systolic blood pressure, ECG abnormalities, diabetes, body mass index, total cholesterol, high-density lipoprotein cholesterol, smoking habits, alcohol intake, and regular exercise (adjusted relative risks, 3.11; 95% CI, 1.04 to 9.32; P=0.04). However, such associations were not observed for ischemic stroke in women or in hemorrhagic stroke in either sex. Among male subjects who were both in the fifth hsCRP level and had hypertension, diabetes, obesity, hypercholesterolemia, or a smoking habit, the risk of ischemic stroke was extremely increased, even after adjustment for other risk factors. CONCLUSIONS: Our findings suggest that elevated serum hsCRP levels are an independent risk factor for future ischemic stroke in Japanese men and that the coexistence of a high hsCRP level with another risk factor extremely increases the risk of ischemic stroke. PMID- 16306469 TI - Incidence and trends of stroke and its subtypes in China: results from three large cities. AB - BACKGROUND AND PURPOSE: To examine the incidence and trends of stroke and its major subtypes during the 1990s in 3 cities in China. METHODS: Stroke cases registered between 1991 to 2000 were initially identified through the stroke surveillance networks established in Beijing, Shanghai, and Changsha, and then confirmed by neurologists. RESULTS: The age-standardized incidence rates per 100,000 person years of overall first-ever stroke were 135.0 (95% CI, 126.5 to 144.6) in Beijing, 76.1 (70.6 to 82.6) in Shanghai, and 150.0 (141.3 to 160.0) in Changsha during the 1990s. Incidence of ischemic stroke (IS) was highest in Beijing, followed by Changsha and Shanghai; for intracerebral hemorrhage (ICH), the highest rate was found in Changsha, followed by Beijing and Shanghai. The same order as ICH was also observed for subarachnoid hemorrhage. The age-adjusted incidence of overall stroke and ICH for individuals > or =55 years of age in our populations was generally higher than that from Western populations. During the 1990s, ICH incidence decreased significantly at a rate of 12.0% per year in Beijing, 4.4% in Shanghai, and 7.7% in Changsha; in contrast, except for Changsha, IS incidence increased in Beijing (5.0% per year) and Shanghai (7.7%). CONCLUSIONS: There is a geographic variation in the incidence of stroke and its subtypes among these 3 cities, but the incidence of overall and hemorrhagic stroke in China is generally higher than that in the Western countries. Interestingly, the decrease in ICH and increase in IS during the past decade may reflect some underlying changes of risk factors in Chinese populations. PMID- 16306470 TI - Fluoxetine treatment in poststroke depression, emotional incontinence, and anger proneness: a double-blind, placebo-controlled study. AB - BACKGROUND AND PURPOSE: The efficacy and safety of the selective serotonin reuptake inhibitor fluoxetine have rarely been studied in the treatment of poststroke emotional disturbances. METHODS: Stroke patients (152) who had poststroke depression (PSD), emotional incontinence (PSEI), or anger proneness (PSAP) were studied. PSD was evaluated by Beck Depression Inventory and Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, PSEI by Kim's criteria, and PSAP was assessed by Spielberger Trait Anger Scale. Subjects were randomly given either fluoxetine 20 mg/day (n=76) or placebo (n=76) for 3 months. Follow-up evaluations were done 1, 3, and 6 months after the beginning of the treatment. The primary outcome measurement was the scores of emotional disturbances at each follow-up assessment. The secondary outcome measurements were the percentage changes of the scores and the subjective responses of the patients. RESULTS: Although patients in the fluoxetine group more often dropped out because of adverse effects, fluoxetine administration was generally safe. Fluoxetine significantly improved PSEI and PSAP, whereas no definitive improvement of PSD was found. Improvement of PSAP was noted even at 3 months after the discontinuation of the treatment. CONCLUSIONS: Fluoxetine is efficacious in the treatment of PSEI and PSAP. Its effect on PSD is not solidly confirmed. PMID- 16306475 TI - Looking back: rheumatoid arthritis--aetiology, occurrence and mortality. AB - Kellgren and Lawrence performed the first studies of the incidence and prevalence of rheumatoid arthritis (RA) in the UK. These have since been updated in Norfolk. The current annual incidence is 36/100,0000 in women and 14/100,000 in men. The prevalence is 0.8% of the adult population. The incidence, prevalence and mortality of RA have fallen in women in the last 50 yr. Excess mortality is predominantly due to cardiovascular disease. Risk factors for the development of RA include obesity, smoking, high red meat consumption, a previous blood transfusion and an adverse pregnancy outcome. Protective factors include the oral contraceptive pill and adequate fruit intake. PMID- 16306476 TI - Towards an epidemiology of rheumatoid arthritis outcome with respect to treatment: randomized controlled trials overestimate treatment response and effectiveness. AB - The results and outcomes of randomized clinical trials of leflunomide and anti TNF therapy are much better than are seen in rheumatoid arthritis patients in the community. This appears to be an effect of the clinical trial system. The consequence of deriving effectiveness estimates from clinical trials is to overestimate the effectiveness and thereby the cost-effectiveness of rheumatoid arthritis treatments. PMID- 16306477 TI - A history of the Arthritis Research Campaign Epidemiology Unit, 1954-2004. AB - The founders of the Arthritis Research Campaign (arc) Epidemiology Unit were Kellgren and Lawrence. Their initial work was conducted in miners and then in the general population. They pioneered standardized methods of interview, examination and X-ray evaluation. They also played a key role in the early development of classification criteria for use in epidemiological surveys. The second director, Philip Wood, focused on understanding the impact of rheumatic diseases on the individual and on society. He was largely responsible for the International Classification of Impairments Disabilities and Handicaps. In the last 30 yrs, developments in statistics, computing and genetics have enabled the basic questions of disease aetiology to be re-examined. The third director, Alan Silman, took over in 1988. The Unit now has three research divisions: Genetics and Genomics, Outcome Studies and Population Surveys. PMID- 16306478 TI - Looking back: developments in our understanding of the occurrence, aetiology and prognosis of chronic pain 1954-2004. AB - This article summarizes the work of the Arthritis Research Campaign Epidemiology Unit in the field of pain and soft tissue rheumatism during the 50 years 1954 2004. It reviews the information gathered on the occurrence of symptoms and its causes, which began with Kellgren and Lawrence's pioneering work during the 1950s in the coal fields and general population of Leigh, Greater Manchester. They studied the roles of posture, mechanical load and the physical environment (dampness) on back pain. This was followed by the Calderdale study in the 1980s examining the prevalence of disability and its causes in the population, which demonstrated the important role of regional pain in causing disability. More recent studies in the 1990s and at the beginning of the 21st century have allowed us to define the relative roles of mechanical (load) factors, individual (psychological) factors and the social environment, and the biological mechanisms by which they may result in symptoms. The further challenge over the coming decade is to use our knowledge of the aetiology and influences on outcome to design management strategies which demonstrate improved outcomes for patients. PMID- 16306479 TI - The epidemiology of osteoarthritis: Manchester and beyond. AB - The paper describes the contribution made to the understanding of the epidemiology of osteoarthritis by the Arthritis Research Campaign Epidemiology Unit. Kellgren and Lawrence defined the condition radiographically and produced the widely used Atlas of Standard Radiographs of Arthritis. They documented the age- and sex-specific prevalence of osteoarthritis. Based on studies in occupational groups, they proposed that local biomechanical stress was linked to causation. Philip Wood focused on osteoarthritis as a syndrome of pain and disability. During the directorship of Alan Silman the emphasis has been on osteoarthritis as a public health problem. PMID- 16306480 TI - Looking back: developments in our understanding of the occurrence, aetiology and prognosis of osteoporosis over the last 50 years. AB - This paper summarizes the contribution of the Arthritis Research Campaign Epidemiology Unit, in particular in its role as coordinator of the European Vertebral Osteoporosis Study, to our current understanding of osteoporosis. One in three postmenopausal women has osteoporosis in the UK. The cost to the NHS is around pound 1.7 billion per year. With the ageing population this is destined to rise. PMID- 16306481 TI - Epidemiology of osteoporotic fracture: looking to the future. AB - This paper reviews the recent literature on candidate genes, anthropometric and environmental factors, and the evolving area of intrauterine fetal programming with regard to the development of osteoporosis. PMID- 16306482 TI - An introduction to the Atlas of Standard Radiographs of Arthritis. PMID- 16306483 TI - The Atlas of Standard Radiographs of Arthritis. PMID- 16306484 TI - Looking back: developments in our understanding of the genetic epidemiology of rheumatoid arthritis over the last 50 years. AB - The contribution of the arc Epidemiology Unit over its 50-yr history to the study of the genetic epidemiology of rheumatoid arthritis (RA) has been reviewed. Early family and population based studies carried out by John Lawrence were important in establishing the role of both genetic and environmental factors in determining susceptibility to RA. More recently, under the leadership of Alan Silman, population-based cohorts, twin- and family-based studies have formed the basis for an extensive programme of research aimed at identifying specific genetic factors that might influence susceptibility and outcome. A review of linkage and association studies is presented. PMID- 16306485 TI - Current understanding of the genetic aetiology of rheumatoid arthritis and likely future developments. AB - Most of the work described herein was performed by the North American Rheumatoid Arthritis Consortium (NARAC). After a brief description of the NARAC and the multiplex family resource that has been developed by this consortium, we will summarize the current status of genome-wide screens using this valuable family collection. Next, we describe work that is under way to further delineate the genes on chromosome 18q that demonstrate linkage to rheumatoid arthritis (RA), including an analysis of candidate genes in the region and results of dense association mapping. We also describe an extensive analysis of functional single nucleotide polymorphisms (SNPs) that is under way in collaboration with Celera Diagnostics, as well as studies designed to further dissect the phenotypic and genotypic heterogeneity of RA. We conclude by briefly summarizing our future plans to elucidate the genetic aetiology of RA. PMID- 16306486 TI - Comparison of prostate cancer mortality in five countries: France, Italy, Japan, UK and USA from the WHO mortality database (1960-2000). PMID- 16306487 TI - Lateral plantar nerve injury following steroid injection for plantar fasciitis. AB - A 41 year old man presented with pain and numbness affecting the lateral aspect of his foot after a steroid injection for plantar fasciitis. Examination confirmed numbness and motor impairment of the lateral plantar nerve. The findings were confirmed by electromyographic studies. The anatomy of the lateral plantar nerve and correct technique for injection to treat plantar fasciitis are discussed. PMID- 16306488 TI - Popliteus tendon tenosynovitis. AB - This case report highlights an unusual cause of anterolateral knee pain. The popliteus muscle arises from three origins--that is, the lateral femoral condyle, the fibula head, and the lateral meniscus--and inserts into the proximal tibia above the soleal line. It may be subjected to a number of pathologies including tenosynovitis, acute calcific tendonitis, rupture, and even avulsion. In this case, the diagnosis of popliteus tendon tenosynovitis was not made from magnetic resonance imaging findings, but was confirmed and successfully treated during arthroscopic examination. PMID- 16306489 TI - The lean and slippered pantaloon... PMID- 16306490 TI - Last orders, gents... PMID- 16306491 TI - They think it's all over, but it may not be! PMID- 16306492 TI - 2012 Olympics: who will survive? PMID- 16306493 TI - Yoga for anxiety: a systematic review of the research evidence. AB - Between March and June 2004, a systematic review was carried out of the research evidence on the effectiveness of yoga for the treatment of anxiety and anxiety disorders. Eight studies were reviewed. They reported positive results, although there were many methodological inadequacies. Owing to the diversity of conditions treated and poor quality of most of the studies, it is not possible to say that yoga is effective in treating anxiety or anxiety disorders in general. However, there are encouraging results, particularly with obsessive compulsive disorder. Further well conducted research is necessary which may be most productive if focused on specific anxiety disorders. PMID- 16306494 TI - Evidence for secular trends in children's physical activity behaviour. AB - It is not clear whether the global increase in weight problems in children is the result of excessive energy intake or decreasing energy expenditure. Methodological limitations have made it difficult to analyse. There is evidence that at least part of the problem may lie with increasing energy consumption, but it is important to examine the other side of the energy equation also. However, it is not possible to conclusively describe physical activity trends because of the absence of suitable baseline data. One solution is to summate all available evidence in as many areas of daily activities as possible and then draw tentative conclusions. This review summarises available trend data on direct representations of physical activity in a range of contexts, together with indirect measures such as sedentariness, fitness, and attitudes. The conclusions drawn are: physical activity in clearly defined contexts such as active transport, school physical education, and organised sports is declining in many countries; young people would like to be active but are often constrained by external factors such as school policy or curricula, parental rules in relation to safety and convenience, and physical environmental factors. PMID- 16306495 TI - Frostbite: incidence and predisposing factors in mountaineers. AB - BACKGROUND: Frostbite, the most common cold injury, occurs in mountaineers, a major group at risk, more often than in the general population. OBJECTIVES: To describe the incidence of frostbite and the situations associated with it in mountaineering, emphasising factors that can be modified to decrease its frequency and severity. METHODS: In this cross sectional, questionnaire based study, 637 mountaineers were asked if they had suffered any frostbite injuries during the preceding two years and to provide the personal and circumstantial details. RESULTS: The mean incidence was 366/1000 population per year. Grade 1 injury (83.0%) and hands (26.4%) and feet (24.1%) involvement were most common. There was a significant relation between lack of proper equipment (odds ratio 14.3) or guide (p<0.001) and the injury. Inappropriate clothing, lack or incorrect use of equipment, and lack of knowledge of how to deal with cold and severe weather were claimed to be the main reasons for the injury. CONCLUSIONS: In high altitude and winter expeditions, mountaineers should wear appropriate clothing, have the necessary equipment such as quality boots and mittens, use a competent guide, and have training on how to tackle cold weather. They should also avoid wet clothing, windy terrains, and should never remain in the same position for long periods to reduce the risk of cold injuries. PMID- 16306496 TI - Stress fractures of the femoral shaft in women's college lacrosse: a report of seven cases and a review of the literature. AB - BACKGROUND: Stress fractures do not often occur in the shaft of the femur. They are more common in the femoral neck, the tibial shaft, the metatarsals, and other bones of the foot. In female athletes, stress fractures classically afflict the distance runner, the ballerina, the gymnast, and the figure skater. OBJECTIVES: To describe the clinical presentation, diagnosis, treatment, and outcome of seven college female lacrosse players with femoral shaft stress fractures, and review the literature. RESULTS: The unusual results of this study support the principle that clinical suspicion should be high when treating any female athlete regardless of the sport. In this case series, an abrupt change in the quality of the running surface during the competitive training season was the only underlying common thread among the athletes. CONCLUSION: The findings suggest that risk factors for the female athlete are variable and are no longer limited to the undernourished or overtrained. PMID- 16306497 TI - Classifying sports medicine diagnoses: a comparison of the International classification of diseases 10-Australian modification (ICD-10-AM) and the Orchard sports injury classification system (OSICS-8). AB - BACKGROUND: The International classification of diseases 10-Australian modification (ICD-10-AM) and the Orchard sports injury classification system (OSICS-8) are two classifications currently being used in sports injury research. OBJECTIVES: To compare these two systems to determine which was the more reliable and easier to apply in the classification of injury diagnoses of patients who presented to sports physicians in private sports medicine practice. METHODS: Ten sports physicians/sports physician registrars each coded one of 10 different lists of 30 sports medicine diagnoses according to both ICD-10-AM and OSICS-8 in random order. The coders noted the time taken to apply each classification system, and allocated an ease of fit score for individual diagnoses into the systems. The 300 diagnoses were each coded twice more by "expert" coders from each system, and these results compared with those of the 10 volunteers. RESULTS: Overall, there was a higher level of agreement between the different coders for OSICS-8 than for ICD-10-AM. On average, it was 23.5 minutes quicker to complete the task with OSICS-8 than with ICD-10-AM. Furthermore, there was also higher concordance between the three coders with OSICS-8. Subjective analysis of the codes assigned indicated reasons for disagreement and showed that, in some instances, even the "expert" coders had difficulties in assigning the most appropriate codes. CONCLUSIONS: Based on the results of this study, OSICS-8 appears to be the preferred system for use by inexperienced coders in sports medicine research. The agreement between coders was, however, lower than expected. It is recommended that changes be made to both OSICS-8 and ICD-10-AM to improve their reliability for use in sports medicine research. PMID- 16306498 TI - How evidence based is the management of two common sports injuries in a sports injury clinic? AB - OBJECTIVES: To examine the diagnosis and management of adults attending a sports injury clinic, to establish to what extent the management of the two most common injuries treated at this clinic is evidence based, and to explore factors that affect management. METHODS: A retrospective examination of 100 random case notes extracted age, sex, sport, type and site of injury, treatment, and outcome. Systematic literature reviews examined the extent and quality of scientific evidence for the management of the two most commonly presenting injuries. A clinical attachment period and practitioner interviews allowed recognition of factors impinging on management decisions. RESULTS: Patellofemoral pain syndrome (PFPS; 10% of all injuries) and Achilles tendinopathy (6% of all injuries) were the most commonly presenting injuries. The mean (SD) number of treatments used for PFPS was 2.8 (0.9). The mean number of treatments used for Achilles tendinopathy was 3.7 (1.0). Clinicians reported that personal experience formed the basis of management plans in 44% of PFPS cases and 59% of Achilles tendinopathy cases, and that primary research evidence only accounted for 24% of management plans in PFPS and 14% in Achilles tendinopathy. Practitioners were unaware of literature supporting over 50% of the treatment modalities they used. However, clinicians were often using evidence based treatments, unaware of the supporting research data. CONCLUSIONS: This study highlights a lack of evidence base, a lack of knowledge of the research evidence, and a lack of management based on the current evidence that is available for these conditions. Practitioners practised evidence based medicine in under 50% of cases. PMID- 16306499 TI - Effects of inhaled bronchodilators and corticosteroids on exercise induced arterial hypoxaemia in trained male athletes. AB - OBJECTIVES: To determine the effect of prophylactic treatment with an inhaled bronchodilator and anti-inflammatory on arterial saturation (SaO2) in trained non asthmatic male athletes with exercise induced arterial hypoxaemia (EIAH). METHODS: Nine male athletes (mean (SD) age 26.3 (6.7) years, height 182.6 (7.9) cm, weight 79.3 (10.5) kg, VO2MAX 62.3 (6.3) ml/kg/min, SaO2MIN 92.5 (1.1)%) with no history of asthma were tested in two experimental conditions. A combination of a therapeutic dose of salbutamol and fluticasone or an inert placebo was administered in a randomised crossover design for seven days before maximal cycling exercise. Oxygen consumption (VO2), ventilation (VE), heart rate (HR), power output, and SaO2 were monitored during the exercise tests. RESULTS: There were no significant differences between the drug (D) and placebo (P) conditions for minimal SaO2 (D = 93.6 (1.4), P = 93.0 (1.1)%; p = 0.93) VO2MAX (D = 61.5 (7.2), P = 61.9 (6.3) ml/kg/min; p = 0.91), peak power (D = 444.4 (48.3), P = 449.4 (43.9) W; p = 0.90), peak VE (D = 147.8 (19.1), P = 149.2 (15.5) litres/min; p = 0.82), or peak heart rate (D = 182.3 (10.0), P = 180.8 (5.5) beats/min; p = 0.76). CONCLUSIONS: A therapeutic dose of salbutamol and fluticasone did not attenuate EIAH during maximal cycling in a group of trained male non-asthmatic athletes. PMID- 16306500 TI - Specific incremental test in elite squash players. AB - OBJECTIVES: To compare cardiorespiratory responses between incremental treadmill (non-specific) and field (sport specific) tests in elite squash players. METHODS: Seven elite players (ranked 1 to 25 in their national federation including the World number 1) randomly performed an incremental treadmill test (TT) and a squash specific graded test (ST) to exhaustion. The ST consisted of repeated displacements replicating the game of squash, at increasing speed on the court. In both tests, ventilatory variables and heart rate were determined at the ventilatory threshold, respiratory compensation point, and maximal loads (max). RESULTS: Heart rate and percentage maximal oxygen uptake (VO2MAX) at the ventilatory threshold and respiratory compensation point were not different between the ST and TT, whereas VO2MAX was higher in the ST than in the TT (63.6 (3.0) v 54.9 (2.5) ml/kg/min; p < 0.001). Time to exhaustion was not different between the ST and TT (1056 (180) v 962 (71) seconds) but correlated with the ranking of the players only in the ST (r = -0.96, p < 0.001). CONCLUSIONS: VO2MAX values derived from laboratory testing were not relevant for accurately estimating fitness in elite squash players. So the ST may be used as an additional test for determination of training intensity. Improved training advice for prescribing aerobic exercise or perfecting stroke technique may result from these results. PMID- 16306501 TI - Physiological characteristics of top level off-road motorcyclists. AB - OBJECTIVES: The study aims to analyse the physiological characteristics of top level off-road motocross, enduro, and desert rally motorcyclists to facilitate the design of a specific training program. METHODS: Twenty seven off-road top level riders participated in this study which measured anthropometric data, maximum aerobic power with incremental tests of both arms and legs, isokinetic dynamometry of the knee and elbow, handgrip strength, heart rate, and blood lactate concentrations during competition. RESULTS: The physical demands of the various races appear to influence the development of distinct musculoskeletal characteristics, as well as aerobic and anaerobic metabolism. Motocross riders have more muscle mass, higher isokinetic handgrip strength, and greater aerobic power than enduro and desert rally riders. However, there are no significant anthropometric and physiological differences between desert rally and enduro riders. Desert rally riders tend to be overweight with maximum aerobic power similar to that of healthy individuals. The mechanical characteristics of the motorcycle and the technical and tactical skills of the riders seem to be more important for race success than the metabolic capabilities of the rider. CONCLUSIONS: Desert rally and enduro riders present similar anthropometric and physiological characteristics. Both have a maximum aerobic power similar to that of healthy normal individuals, although desert rally riders tend to be overweight. Motocross riders on the other hand, have more muscle mass, more strength, and greater aerobic power. The differences observed suggest the need for a specific training program to address the requirements of different riders to reduce the possibility of injury. PMID- 16306502 TI - Neuromuscular and biomechanical characteristic changes in high school athletes: a plyometric versus basic resistance program. AB - BACKGROUND: In order to improve neuromuscular and biomechanical characteristic deficits in female athletes, numerous injury prevention programs have been developed and have successfully reduced the number of knee ligament injuries. However, few have investigated the neuromuscular and biomechanical changes following these training programs. It is also largely unknown what type of program is better for improving the landing mechanics of female athletes. OBJECTIVES: To investigate the effects of an 8 week plyometric and basic resistance training program on neuromuscular and biomechanical characteristics in female athletes. METHODS: Twenty seven high school female athletes participated either in a plyometric or a basic resistance training program. Knee and hip strength, landing mechanics, and muscle activity were recorded before and after the intervention programs. In the jump-landing task, subjects jumped as high as they could and landed on both feet. Electromyography (EMG) peak activation time and integrated EMG of thigh and hip muscles were recorded prior to (preactive) and subsequent to (reactive) foot contact. RESULTS: Both groups improved knee extensor isokinetic strength and increased initial and peak knee and hip flexion, and time to peak knee flexion during the task. The peak preactive EMG of the gluteus medius and integrated EMG for the gluteus medius during the preactive and reactive time periods were significantly greater for both groups. CONCLUSIONS: Basic training alone induced favourable neuromuscular and biomechanical changes in high school female athletes. The plyometric program may further be utilised to improve muscular activation patterns. PMID- 16306503 TI - Initial effects of anti-pronation tape on the medial longitudinal arch during walking and running. AB - OBJECTIVES: To investigate the effect of an augmented LowDye taping technique on the medial longitudinal arch of the foot during dynamic tasks such as walking and jogging, and to elucidate the relation between tape induced changes in static and dynamic foot posture. METHODS: Seventeen subjects (mean (SD) age 27 (5.8) years) who were asymptomatic and exhibited a navicular drop greater than 10 mm were studied. Medial longitudinal arch height standardised to foot length during standing and at mid-stance of walking and jogging was measured from digital video images taken before and after the application of an anti-pronation taping technique. A no tape control condition was also included. RESULTS: Compared with the no tape control condition, tape produced a significant mean (SD) increase in the medial longitudinal arch height index of 0.031 (0.015), 0.026 (0.014), and 0.016 (0.017) during standing, walking, and jogging respectively (p < 0.05). The relative increase in medial longitudinal arch height represents an anti-pronation effect. The tape induced changes in the medial longitudinal arch height measured during standing correlated strongly with those measured during walking and jogging (Pearson's r = 0.7 and 0.76 respectively). CONCLUSIONS: The augmented LowDye tape was effective in controlling pronation during both static and dynamic activity. Tape induced changes in static foot posture paralleled those during walking and jogging. PMID- 16306504 TI - An exercise programme for the management of lateral elbow tendinopathy. AB - BACKGROUND: Home exercise programmes and exercise programmes carried out in a clinical setting are commonly advocated for the treatment of lateral elbow tendinopathy (LET), a very common lesion of the arm with a well-defined clinical presentation. The aim of this study is to describe the use and effects of strengthening and stretching exercise programmes in the treatment of LET. ECCENTRIC EXERCISES: Slow progressive eccentric exercises for LET should be performed with the elbow in extension, forearm in pronation, and wrist in extended position (as high as possible). However, it is unclear how the injured tendon, which is loaded eccentrically, returns to the starting position without experiencing concentric loading and how the "slowness" of eccentric exercises should be defined. Nor has the treatment regimen of the eccentric exercises of a supervised exercise programme been defined. STRETCHING EXERCISES: Static stretching is defined as passively stretching a given muscle-tendon unit by slowly placing and maintaining it in a maximal position of stretch. We recommend the position should be held for 30-45 s, three times before and three times after eccentric exercises during each treatment session with a 30 s rest interval between each procedure. The treatment region of static stretching exercises when a supervised exercise programme is performed is unknown. DISCUSSION: A well designed trial is needed to study the effectiveness of a supervised exercise programme for LET consisting of eccentric and static stretching exercises. The issues relating to the supervised exercise programme should be defined so that therapists can replicate the programme. PMID- 16306505 TI - Effects of dietary carbohydrate on delayed onset muscle soreness and reactive oxygen species after contraction induced muscle damage. AB - BACKGROUND: Delayed onset muscle soreness (DOMS) occurs after unaccustomed exercise and has been suggested to be attributable to reactive oxygen species (ROS). Previous studies have shown increased ROS after lengthening contractions, attributable to invading phagocytes. Plasma glucose is a vital fuel for phagocytes, therefore carbohydrate (CHO) status before exercise may influence ROS production and DOMS. OBJECTIVE: To examine the effect of pre-exercise CHO status on DOMS, ROS production, and muscle function after contraction induced muscle damage. METHOD: Twelve subjects performed two downhill runs, one after a high CHO diet and one after a low CHO diet. Blood samples were drawn for analysis of malondialdehyde, total glutathione, creatine kinase, non-esterified fatty acids, lactate, glucose, and leucocytes. DOMS and muscle function were assessed daily. RESULTS: The high CHO diet resulted in higher respiratory exchange ratio and lactate concentrations than the low CHO diet before exercise. The low CHO diet resulted in higher non-esterified fatty acid concentrations before exercise. DOMS developed after exercise and remained for up to 96 hours, after both diets. A biphasic response in creatine kinase occurred after both diets at 24 and 96 hours after exercise. Malondialdehyde had increased 72 hours after exercise after both diets, and muscle function was attenuated up to this time. CONCLUSIONS: Downhill running resulted in increased ROS production and ratings of DOMS and secondary increases in muscle damage. CHO status before exercise had no effect. PMID- 16306506 TI - Effects of dominant somatotype on aerobic capacity trainability. AB - PURPOSE: This study examined the association between dominant somatotype and the effect on aerobic capacity variables of individualised aerobic interval training. METHODS: Forty one white North African subjects (age 21.4+/-1.3 years; Vo2max = 52.8+/-5.7 ml kg(-1) min(-1)) performed three exercise tests 1 week apart (i) an incremental test on a cycle ergometer to determine Vo2max and Vo2 at the second ventilatory threshold (VT2); (ii) a VAM-EVAL track test to determine maximal aerobic speed (vVo2max); and (iii) an exhaustive constant velocity test to determine time limit performed at 100% vVo2max (tlim100). Subjects were divided into four somatometric groups: endomorphs-mesomorphs (Endo-meso; n = 9), mesomorphs (Meso; n = 11), mesomorphs-ectomorphs (Meso-ecto; n = 12), and ectomorphs (Ecto; n = 9). Subjects followed a 12 week training program (two sessions/week). Each endurance training session consisted of the maximal number of successive fractions for each subject. Each fraction consisted of one period of exercise at 100% of vVo2max and one of active recovery at 60% of vVo2max. The duration of each period was equal to half the individual tlim100 duration (153.6+/-39.7 s). After the training program, all subjects were re-evaluated for comparison with pre-test results. RESULTS: Pre- and post-training data were grouped by dominant somatotype. Two way ANOVA revealed significant somatotype aerobic training interaction effects (p<0.001) for improvements in vVo2max, Vo2max expressed classically and according to allometric scaling, and Vo2 at VT2. There were significant differences among groups post-training: the Meso-ecto and the Meso groups showed the greatest improvements in aerobic capacity. CONCLUSION: The significant somatotype-aerobic training interaction suggests different trainability with intermittent and individualised aerobic training according to somatotype. PMID- 16306507 TI - Effect of swimming intensity on subsequent cycling and overall triathlon performance. AB - OBJECTIVES: To investigate the effects of different swimming intensities on subsequent cycling and overall triathlon performance. METHODS: Nine highly trained, male triathletes completed five separate laboratory sessions comprising one graded exercise test, a swim time trial (STT), and three sprint distance triathlons (TRI). The swimming velocities of the three TRI sessions were 80-85% (S80), 90-95% (S90), and 98-102% (S100) of the STT velocity. Subsequent cycling and running were performed at a perceived maximal intensity. Swimming stroke mechanics were measured during the swim. Plasma lactate concentration and ratings of perceived exertion were recorded at the conclusion of the swim and over the course of subsequent cycling and running. Oxygen consumption was recorded during the cycle. RESULTS: The S80 and S90 cycle times were faster than the S100 cycle time (p<0.05). The overall triathlon time of S80 was faster than that of S100 (p<0.05). The S100 swim was characterised by a greater stroke rate than S80 and S90 (p<0.05) and a greater plasma lactate concentration than S80 (p<0.01). CONCLUSION: A swimming intensity below that of a time trial effort significantly improves subsequent cycling and overall triathlon performance. PMID- 16306508 TI - Specificity of VO2MAX and the ventilatory threshold in free swimming and cycle ergometry: comparison between triathletes and swimmers. AB - OBJECTIVES: To compare maximal heart rate (HRmax), maximal oxygen consumption (VO2MAX), and the ventilatory threshold (VT; %VO2MAX) during cycle ergometry and free swimming between swimmers and triathletes. METHODS: Nine swimmers and ten triathletes completed an incremental swimming and cycling test to exhaustion. Whole body metabolic responses were determined in each test. RESULTS: The swimmers exhibited a significantly higher VO2MAX in swimming than in cycling (58.4 (5.6) v 51.3 (5.1) ml/kg/min), whereas the opposite was found in the triathletes (53.0 (6.7) v 68.2 (6.8) ml/kg/min). HRmax was significantly different in the maximal cycling and swimming tests for the triathletes (188.6 (7.5) v 174.8 (9.0) beats/min). In the maximal swimming test, HRmax was significantly higher in the swimmers than in the triathletes (174.8 (9.0) v 184.6 (9.7) beats/min). No significant differences were found for VT measured in swimming and cycling in the triathletes and swimmers. CONCLUSION: This study confirms that the exercise testing mode affects the VO2MAX value, and that swimmers have very specific training adaptations even compared with triathletes. This may be a function of acute physiological responses combined with the specialist training status of the different athletes influencing maximal cardiac output or oxygen extraction. In contrast, the different training regimens do not seem to influence the VT, as this variable did not differ between the two testing modes in either group. PMID- 16306509 TI - William Penny Brookes (1809-1895): forgotten Olympic Lord of the Rings. AB - The contribution of Dr William Penny Brookes (1809-1895) to the Olympic Games is scarcely reported in the medical literature. Presented here is a brief report of his life and work. Furthermore, the need to promote public health and fitness through the Olympics is re-emphasised. PMID- 16306512 TI - Shortchanged by Sir2. PMID- 16306513 TI - Mitochondrial Dynamics in Cell Life and Death. AB - Mitochondria are dynamic organelles that are able to change their morphology during cellular development and to meet the energetic needs of the cell, for example. During the past several years, knowledge about the molecular machinery involved in the control of mitochondrial shape has improved considerably. In August 2005, leading researchers in the field of mitochondrial dynamics met at the Venetian Institute of Molecular Medicine in Padova, Italy, to discuss novel and intriguing findings presented at the first workshop on "Mitochondrial Dynamics in Cell Life and Death." Some of the findings related to apoptosis and aging are described in this Perspective. PMID- 16306514 TI - Images in clinical medicine. Caput medusae. PMID- 16306515 TI - The origins of pandemic influenza--lessons from the 1918 virus. PMID- 16306516 TI - Biomedical research and biosecurity. PMID- 16306517 TI - Hostile use of the life sciences. PMID- 16306518 TI - Neuroblastoma--from genetic profiles to clinical challenge. PMID- 16306519 TI - Mycophenolate mofetil or intravenous cyclophosphamide for lupus nephritis. AB - BACKGROUND: Since anecdotal series and small, prospective, controlled trials suggest that mycophenolate mofetil may be effective for treating lupus nephritis, larger trials are desirable. METHODS: We conducted a 24-week randomized, open label, noninferiority trial comparing oral mycophenolate mofetil (initial dose, 1000 mg per day, increased to 3000 mg per day) with monthly intravenous cyclophosphamide (0.5 g per square meter of body-surface area, increased to 1.0 g per square meter) as induction therapy for active lupus nephritis. A change to the alternative regimen was allowed at 12 weeks in patients who did not have an early response. The study protocol specified adjunctive care and the use and tapering of corticosteroids. The primary end point was complete remission at 24 weeks (normalization of abnormal renal measurements and maintenance of baseline normal measurements). A secondary end point was partial remission at 24 weeks. RESULTS: Of 140 patients recruited, 71 were randomly assigned to receive mycophenolate mofetil and 69 were randomly assigned to receive cyclophosphamide. At 12 weeks, 56 patients receiving mycophenolate mofetil and 42 receiving cyclophosphamide had satisfactory early responses. In the intention-to-treat analysis, 16 of the 71 patients (22.5 percent) receiving mycophenolate mofetil and 4 of the 69 patients receiving cyclophosphamide (5.8 percent) had complete remission, for an absolute difference of 16.7 percentage points (95 percent confidence interval, 5.6 to 27.9 percentage points; P=0.005), meeting the prespecified criteria for noninferiority and demonstrating the superiority of mycophenolate mofetil to cyclophosphamide. Partial remission occurred in 21 of the 71 patients (29.6 percent) and 17 of the 69 patients (24.6 percent), respectively (P=0.51). Three patients assigned to cyclophosphamide died, two during protocol therapy. Fewer severe infections and hospitalizations but more diarrhea occurred among those receiving mycophenolate. CONCLUSIONS: In this 24 week trial, mycophenolate mofetil was more effective than intravenous cyclophosphamide in inducing remission of lupus nephritis and had a more favorable safety profile. PMID- 16306520 TI - High-dose acetylcysteine in idiopathic pulmonary fibrosis. AB - BACKGROUND: Idiopathic pulmonary fibrosis is a chronic progressive disorder with a poor prognosis. METHODS: We conducted a double-blind, randomized, placebo controlled multicenter study that assessed the effectiveness over one year of a high oral dose of acetylcysteine (600 mg three times daily) added to standard therapy with prednisone plus azathioprine. The primary end points were changes between baseline and month 12 in vital capacity and in single-breath carbon monoxide diffusing capacity (DL(CO)). RESULTS: A total of 182 patients were randomly assigned to treatment (92 to acetylcysteine and 90 to placebo). Of these patients, 155 (80 assigned to acetylcysteine and 75 to placebo) had usual interstitial pneumonia, as confirmed by high-resolution computed tomography and histologic findings reviewed by expert committees, and did not withdraw consent before the start of treatment. Fifty-seven of the 80 patients taking acetylcysteine (71 percent) and 51 of the 75 patients taking placebo (68 percent) completed one year of treatment. Acetylcysteine slowed the deterioration of vital capacity and DL(CO): at 12 months, the absolute differences in the change from baseline between patients taking acetylcysteine and those taking placebo were 0.18 liter (95 percent confidence interval, 0.03 to 0.32), or a relative difference of 9 percent, for vital capacity (P=0.02), and 0.75 mmol per minute per kilopascal (95 percent confidence interval, 0.27 to 1.23), or 24 percent, for DL(CO) (P=0.003). Mortality during the study was 9 percent among patients taking acetylcysteine and 11 percent among those taking placebo (P=0.69). There were no significant differences in the type or severity of adverse events between patients taking acetylcysteine and those taking placebo, except for a significantly lower rate of myelotoxic effects in the group taking acetylcysteine (P=0.03). CONCLUSIONS: Therapy with acetylcysteine at a dose of 600 mg three times daily, added to prednisone and azathioprine, preserves vital capacity and DL(CO) in patients with idiopathic pulmonary fibrosis better than does standard therapy alone. PMID- 16306521 TI - Chromosome 1p and 11q deletions and outcome in neuroblastoma. AB - BACKGROUND: Neuroblastoma is a childhood cancer with considerable morbidity and mortality. Tumor-derived biomarkers may improve risk stratification. METHODS: We screened 915 samples of neuroblastoma for loss of heterozygosity (LOH) at chromosome bands 1p36 and 11q23. Additional analyses identified a subgroup of cases of 11q23 LOH with unbalanced 11q LOH (unb11q LOH; defined as loss of 11q with retention of 11p). The associations of LOH with relapse and survival were determined. RESULTS: LOH at 1p36 was identified in 209 of 898 tumors (23 percent) and LOH at 11q23 in 307 of 913 (34 percent). Unb11q LOH was found in 151 of 307 tumors with 11q23 LOH (17 percent of the total cohort). There was a strong association of 1p36 LOH, 11q23 LOH, and unb11q LOH with most high-risk disease features (P<0.001). LOH at 1p36 was associated with amplification of the MYCN oncogene (P<0.001), but 11q23 LOH and unb11q LOH were not (P<0.001 and P=0.002, respectively). Cases with unb11q LOH were associated with three-year event-free and overall survival rates (+/-SE) of 50+/-5 percent and 66+/-5 percent, respectively, as compared with 74+/-2 percent and 83+/-2 percent among cases without unb11q LOH (P<0.001 for both comparisons). In a multivariate model, unb11q LOH was independently associated with decreased event-free survival (P=0.009) in the entire cohort, and both 1p36 LOH and unb11q LOH were independently associated with decreased progression-free survival in the subgroup of patients with features of low-risk and intermediate-risk disease (P=0.002 and P=0.02, respectively). CONCLUSIONS: Unb11q LOH and 1p36 LOH are independently associated with a worse outcome in patients with neuroblastoma. PMID- 16306522 TI - Beta-blockers to prevent gastroesophageal varices in patients with cirrhosis. AB - BACKGROUND: Nonselective beta-adrenergic blockers decrease portal pressure and prevent variceal hemorrhage. Their effectiveness in preventing varices is unknown. METHODS: We randomly assigned 213 patients with cirrhosis and portal hypertension (minimal hepatic venous pressure gradient [HVPG] of 6 mm Hg) to receive timolol, a nonselective beta-blocker (108 patients), or placebo (105 patients). The primary end point was the development of gastroesophageal varices or variceal hemorrhage. Endoscopy and HVPG measurements were repeated yearly. RESULTS: During a median follow-up of 54.9 months, the rate of the primary end point did not differ significantly between the timolol group and the placebo group (39 percent and 40 percent, respectively; P=0.89), nor were there significant differences in the rates of ascites, encephalopathy, liver transplantation, or death. Serious adverse events were more common among patients in the timolol group than among those in the placebo group (18 percent vs. 6 percent, P=0.006). Varices developed less frequently among patients with a baseline HVPG of less than 10 mm Hg and among those in whom the HVPG decreased by more than 10 percent at one year and more frequently among those in whom the HVPG increased by more than 10 percent at one year. CONCLUSIONS: Nonselective beta blockers are ineffective in preventing varices in unselected patients with cirrhosis and portal hypertension and are associated with an increased number of adverse events. (ClinicalTrials.gov number, NCT00006398.) PMID- 16306523 TI - Basal-cell carcinoma. PMID- 16306524 TI - Images in clinical medicine. Unfortunate ingestion. PMID- 16306525 TI - Case records of the Massachusetts General Hospital. Case 36-2005. A 61-year-old woman with seizure, disturbed gait, and altered mental status. PMID- 16306526 TI - Mycophenolate mofetil for lupus nephritis. PMID- 16306527 TI - Antioxidant therapy for idiopathic pulmonary fibrosis. PMID- 16306528 TI - Beta-blockers to prevent esophageal varices--an unfulfilled promise. PMID- 16306529 TI - Jumping frogs, endangered toads, and California's medical-marijuana law. PMID- 16306530 TI - Inducing intestinal growth. PMID- 16306531 TI - Hepatitis A associated with green onions. PMID- 16306532 TI - Nomogram for exercise capacity in women. PMID- 16306533 TI - Soft-tissue sarcoma. PMID- 16306534 TI - Case 24-2005: a woman with estrogen-receptor-positive breast cancer. PMID- 16306535 TI - Mitochondrial toxicity associated with linezolid. PMID- 16306536 TI - Refeeding David Blaine--studies after a 44-day fast. PMID- 16306537 TI - A kingpin of academic inclusive fitness: the history and contributions of Bruce Grant. PMID- 16306538 TI - Effective population size under random mating with a finite number of matings. PMID- 16306539 TI - Decreasing the risk of diabetic retinopathy in a study of case management: the California Medi-Cal Type 2 Diabetes Study. AB - OBJECTIVE: Diabetic retinopathy affects >60% of people with type 2 diabetes during the first 2 decades of the disease and is ameliorated by good glycemic control. This study tested whether intensive diabetes case management could prevent or delay diabetic retinopathy in patients with established type 2 diabetes. RESEARCH DESIGN AND METHODS: This study was part of a randomized, controlled clinical trial of diabetes case management in type 2 diabetes in southern California counties serving low income ethnic minority populations. Subjects were randomized to intervention (diabetes case management) or control (traditional treatment) groups. Subjects with at least two retinal photographs (n = 149) were included in this analysis to assess the effect of intervention on development or progression of diabetic retinopathy. RESULTS: Progression of retinopathy in the intervention group was not significantly less than in the control group (P = 0.226). However, those in the intervention group with no evidence of retinopathy at baseline were less likely to develop diabetic retinal changes (5/48) during a mean follow-up of 23.1 months than those in the control group (10/34, chi(2) = 4.805, P = 0.028). This difference remained significant in a logistic regression model that controlled for potential confounders (odds ratio 5.35 [95% CI 1.14-25.12]). CONCLUSIONS: This study shows that a relatively short duration of case management instituted before the onset of clinically identifiable retinopathy significantly diminished the risk of developing retinopathy in patients with type 2 diabetes. The findings also emphasized the retinal disease burden in this population, with development and progression of retinopathy occurring in <2 years. PMID- 16306540 TI - Beneficial effects of a Dietary Approaches to Stop Hypertension eating plan on features of the metabolic syndrome. AB - OBJECTIVE: To determine the effects of a Dietary Approaches to Stop Hypertension (DASH) eating plan on metabolic risks in patients with the metabolic syndrome. RESEARCH DESIGN AND METHODS: This was a randomized controlled outpatient trial conducted on 116 patients with the metabolic syndrome. Three diets were prescribed for 6 months: a control diet, a weight-reducing diet emphasizing healthy food choices, and the DASH diet with reduced calories and increased consumption of fruit, vegetables, low-fat dairy, and whole grains and lower in saturated fat, total fat, and cholesterol and restricted to 2,400 mg Na. The main outcome measures were the components of the metabolic syndrome. RESULTS: Relative to the control diet, the DASH diet resulted in higher HDL cholesterol (7 and 10 mg/dl), lower triglycerides (-18 and -14 mg/dl), systolic blood pressure (SBP) ( 12 and -11 mmHg), diastolic blood pressure (-6 and -7 mmHg), weight (-16 and -14 kg), fasting blood glucose (FBG) (-15 and -8 mg/dl), and weight (-16 and -15 kg), among men and women, respectively (all P < 0.001). The net reduction in triglycerides (-17 and -18 mg/dl), SBP (-11 and -11 mmHg), diastolic blood pressure (-5 and -6 mmHg), and FBG (-4 and -6 mg/dl), weight (-16 and -15 kg), and increase in HDL (5 and 10 mg/dl) among men and women, respectively, was higher in the DASH group (all P < 0.05). The weight-reducing diet resulted in significant change in triglycerides (-13 and -10 mg/dl), SBP (-6 and -6 mmHg), and weight (-13 and -12 kg) among men and women, respectively (all P < 0.05). CONCLUSIONS: The DASH diet can likely reduce most of the metabolic risks in both men and women; the related mechanisms need further study. PMID- 16306541 TI - Dietary glycemic index and glycemic load, carbohydrate and fiber intake, and measures of insulin sensitivity, secretion, and adiposity in the Insulin Resistance Atherosclerosis Study. AB - OBJECTIVE: We studied the association of digestible carbohydrates, fiber intake, glycemic index, and glycemic load with insulin sensitivity (S(I)), fasting insulin, acute insulin response (AIR), disposition index, BMI, and waist circumference. RESEARCH DESIGN AND METHODS: Data on 979 adults with normal (67%) and impaired (33%) glucose tolerance from the Insulin Resistance Atherosclerosis Study (1992-1994) were analyzed. Usual dietary intake was assessed via a 114-item interviewer-administered food frequency questionnaire from which nutrient intakes were estimated. Published glycemic index values were assigned to food items and average dietary glycemic index and glycemic load calculated per subject. S(I) and AIR were determined by frequently sampled intravenous glucose tolerance test. Disposition index was calculated by multiplying S(I) with AIR. Multiple linear regression modeling was employed. RESULTS: No association was observed between glycemic index and S(I), fasting insulin, AIR, disposition index, BMI, or waist circumference after adjustment for demographic characteristics or family history of diabetes, energy expenditure, and smoking. Associations observed for digestible carbohydrates and glycemic load, respectively, with S(I), insulin secretion, and adiposity (adjusted for demographics and main confounders) were entirely explained by energy intake. In contrast, fiber was associated positively with S(I) and disposition index and inversely with fasting insulin, BMI, and waist circumference but not with AIR. CONCLUSION: Carbohydrates as reflected in glycemic index and glycemic load may not be related to measures of insulin sensitivity, insulin secretion, and adiposity. Fiber intake may not only have beneficial effects on insulin sensitivity and adiposity, but also on pancreatic functionality. PMID- 16306542 TI - Relative risk of mortality associated with diabetes as a function of birth weight. AB - OBJECTIVE: Birth weight is a risk factor for both diabetes and mortality. Diabetes is a risk factor for mortality. Whether the excess mortality observed for diabetes varies with birth weight is unclear. RESEARCH DESIGN AND METHODS: Among all 2,508 Rochester, Minnesota, residents who first met research criteria for adult-onset diabetes in 1960-1995, 171 were born locally in-hospital after 1922 (i.e., birth weights available) as singleton, term infants. Each case subject and two age- and sex-matched nondiabetic control subjects (born locally, residing locally when the case subject met the criteria for diabetes) were followed through 31 December 2000 for vital status. RESULTS: Of the diabetic case subjects, 16% (27 of 171) died vs. 7% (25 of 342) of control subjects (P = 0.004). The difference was less for normal-birth-weight (NBW) (2,948-<3,856 g) individuals (12% [12 of 102] vs. 8% [20 of 246], P = 0.31) than for abnormal birth-weight individuals (low birth weight [LBW] 20% [8 of 39] vs. 2% [1 of 46], P = 0.01; high birth weight [HBW] 23% [7 of 30] vs. 8% [4 of 50], P = 0.16), as confirmed with age- and sex-adjusted Cox proportional hazards (diabetes associated hazard ratio 1.4 [95% CI 0.69-2.90] for NBW vs. 4.8 [1.7-13.3] for abnormal birth weight, test for interaction P = 0.056). The observed diabetes deaths were greater than expected, based on mortality for the general population (27 vs. 13.3, P < 0.001), with 70% of excess deaths occurring among LBW (8 vs. 2.2, P < 0.001) and HBW (7 vs. 3.1, P = 0.03) individuals. CONCLUSIONS: The excess mortality observed for diabetes appears disproportionately concentrated among abnormal-birth-weight individuals, thus identifying a subset of at-risk diabetic individuals and reinforcing the importance of NBW deliveries. PMID- 16306543 TI - Race differences in long-term diabetes management in an HMO. AB - OBJECTIVE: We examined race differences in diabetes outcomes over 4-8 years in a single HMO. RESEARCH DESIGN AND METHODS: We identified black and white adult diabetic patients who were continuously enrolled (1992-2001) and in whom diabetes was 1) diagnosed before 1994 (n = 1,686) or 2) newly diagnosed in 1994-1997 (n = 1,280). We used hierarchical models to estimate the effect of race on average annual HbA(1c) (A1C) controlling for baseline A1C, BMI, and age, as well as annual measures of type of diabetes medications, diabetes-related hospitalization, time and the number of A1C tests, physician visits, and nondiabetes medications. Stratifying by sex accounted for significant interactions between sex and race. RESULTS: At baseline, black and white patients had similar rates of A1C testing and physician visits, but blacks had higher unadjusted A1C values. In multivariate models, among patients with previously diagnosed diabetes, average A1C was nonsignificantly 0.11 higher (95% CI -0.12 to 0.34) in black than in white men but was 0.30 higher (0.14-0.46; P = 0.0007) in black than in white women. Among patients with newly diagnosed diabetes, the adjusted black-white gap was 0.49 among men (0.17-0.80; P = 0.007) and was 0.05 among women (-0.20 to -0.31), which was positive but not significant. CONCLUSIONS: Factors other than the quality of care may explain persistent race differences in A1C in this setting. Future interventions should target normalization of A1C by identifying potential psychosocial barriers to therapy intensification among patients and clinicians and development of culturally appropriate interventions to aid patients in successful self-management. PMID- 16306544 TI - Elevated hip fracture risk in type 1 diabetic patients: a population-based cohort study in Sweden. AB - OBJECTIVE: Patients with type 1 diabetes often have low bone mineral density, but epidemiological data on fracture risk are sparse and imprecise, particularly for men. RESEARCH DESIGN AND METHODS: In the Swedish Inpatient Register, we identified a population-based cohort of 24,605 patients (12,551 men and 12,054 women) who were hospitalized for diabetes before age 31 years during 1975 through 1998. Follow-up for hip fracture was accomplished through cross-linkage in the Inpatient Register until the end of 1998. Censoring information was obtained from the registers of Death and Migration. Using the Kaplan-Meier method, we calculated the cumulative probability of getting a hip fracture. Standardized hospitalization ratios and their 95% CIs estimated relative risks with the age-, sex-, and calendar period-matched Swedish general population as reference. RESULTS: In total, 70 and 51 first hip fractures were ascertained in men and women, respectively, corresponding to a cumulative probability (both sexes) of 65.8/1,000 until age 65 years. Markedly elevated risks were observed in both men and women (standardized hospitalization ratios = 7.6 [95% CI 5.9-9.6] and 9.8 [7.3-12.9], respectively), increasing with follow-up time. Ophthalmic, nephropathic, neurological, and cardiovascular complications were indicators of particularly high risks. CONCLUSIONS: Both male and female type 1 diabetic patients are at increased risk for hip fracture. Although optimal preventive measures still need to be defined, the co-occurrence with other diabetes complications suggests that tighter metabolic control might reduce the risk. PMID- 16306545 TI - A computer simulation model of diabetes progression, quality of life, and cost. AB - OBJECTIVE: To develop and validate a comprehensive computer simulation model to assess the impact of screening, prevention, and treatment strategies on type 2 diabetes and its complications, comorbidities, quality of life, and cost. RESEARCH DESIGN AND METHODS: The incidence of type 2 diabetes and its complications and comorbidities were derived from population-based epidemiologic studies and randomized, controlled clinical trials. Health utility scores were derived for patients with type 2 diabetes using the Quality of Well Being-Self Administered. Direct medical costs were derived for managed care patients with type 2 diabetes using paid insurance claims. Monte Carlo techniques were used to implement a semi-Markov model. Performance of the model was assessed using baseline and 4- and 10-year follow-up data from the older-onset diabetic population studied in the Wisconsin Epidemiologic Study of Diabetic Retinopathy (WESDR). RESULTS: Applying the model to the baseline WESDR population with type 2 diabetes, we predicted mortality to be 51% at 10 years. The prevalences of stroke and myocardial infarction were predicted to be 18 and 19% at 10 years. The prevalences of nonproliferative diabetic retinopathy, proliferative retinopathy, and macular edema were predicted to be 45, 16, and 18%, respectively; the prevalences of microalbuminuria, proteinuria, and end-stage renal disease were predicted to be 19, 39, and 3%, respectively; and the prevalences of clinical neuropathy and amputation were predicted to be 52 and 5%, respectively, at 10 years. Over 10 years, average undiscounted total direct medical costs were estimated to be USD $53,000 per person. Among survivors, the average utility score was estimated to be 0.56 at 10 years. CONCLUSIONS: Our computer simulation model accurately predicted survival and the cardiovascular, microvascular, and neuropathic complications observed in the WESDR cohort with type 2 diabetes over 10 years. The model can be used to predict the progression of diabetes and its complications, comorbidities, quality of life, and cost and to assess the relative effectiveness, cost-effectiveness, and cost-utility of alternative strategies for the prevention and treatment of type 2 diabetes. PMID- 16306546 TI - Race, ethnicity, socioeconomic position, and quality of care for adults with diabetes enrolled in managed care: the Translating Research Into Action for Diabetes (TRIAD) study. AB - OBJECTIVE: To examine racial/ethnic and socioeconomic variation in diabetes care in managed-care settings. RESEARCH DESIGN AND METHODS: We studied 7,456 adults enrolled in health plans participating in the Translating Research Into Action for Diabetes study, a six-center cohort study of diabetes in managed care. Cross sectional analyses using hierarchical regression models assessed processes of care (HbA(1c) [A1C], lipid, and proteinuria assessment; foot and dilated eye examinations; use or advice to use aspirin; and influenza vaccination) and intermediate health outcomes (A1C, LDL, and blood pressure control). RESULTS: Most quality indicators and intermediate outcomes were comparable across race/ethnicity and socioeconomic position (SEP). Latinos and Asians/Pacific Islanders had similar or better processes and intermediate outcomes than whites with the exception of slightly higher A1C levels. Compared with whites, African Americans had lower rates of A1C and LDL measurement and influenza vaccination, higher rates of foot and dilated eye examinations, and the poorest blood pressure and lipid control. The main SEP difference was lower rates of dilated eye examinations among poorer and less educated individuals. In almost all instances, racial/ethnic minorities or low SEP participants with poor glycemic, blood pressure, and lipid control received similar or more appropriate intensification of therapy relative to whites or those with higher SEP. CONCLUSIONS: In these managed-care settings, minority race/ethnicity was not consistently associated with worse processes or outcomes, and not all differences favored whites. The only notable SEP disparity was in rates of dilated eye examinations. Social disparities in health may be reduced in managed-care settings. PMID- 16306547 TI - Comparison of a needle-type and a microdialysis continuous glucose monitor in type 1 diabetic patients. AB - OBJECTIVE: We examined the reliability of two continuous glucose sensors in type 1 diabetic patients at night and during rapid glucose excursions and verified the hypothesized nocturnal hypoglycemic drift of the needle-type sensor (CGMSgold) and delay of the microdialysis sensor (GlucoDay). RESEARCH DESIGN AND METHODS: Blood was sampled overnight twice per hour in 13 patients. Rapid-acting insulin was given subcutaneously 30 min after breakfast. Sampling once per minute started 45 min after breakfast and 75 min after insulin injection for 30 min, with the aim of determining peak and nadir glucose values. Mean absolute differences (MADs) between sensor and blood glucose values were calculated. Sensor curves were modeled for all patients using linear regression. Horizontal and vertical shifts of sensor curves from the blood glucose curves were assessed. A vertical shift indicates sensor drift and a horizontal shift sensor delay. RESULTS: Drift was minimal in the needle-type and microdialysis sensors (-0.02 and -0.04 mmol/l). Mean +/- SD delay was 7.1 +/- 5.5 min for the microdialysis sensor (P < 0.001). MAD was 15.0% for the needle-type sensor and 13.6% for the microdialysis sensor (P = 0.013). After correction for the 7-min delay, the microdialysis MAD improved to 11.7% (P < 0.0001). CONCLUSIONS: The microdialysis sensor was more accurate than the needle-type sensor, with or without correction for a 7-min delay. In contrast to the previous version, the current needle-type sensor did not exhibit nocturnal hypoglycemic drift. Continuous subcutaneous glucose sensors are valuable adjunctive tools for glucose trend analyses. However, considering the large MADs, individual sensor values should be interpreted with caution. PMID- 16306548 TI - Rosiglitazone improves exercise capacity in individuals with type 2 diabetes. AB - OBJECTIVE: Although exercise is recommended as a cornerstone of treatment for type 2 diabetes, it is often poorly adopted by patients. We have noted that even in the absence of apparent cardiovascular disease, persons with type 2 diabetes have an impaired ability to carry out maximal exercise, and the impairment is correlated with insulin resistance and endothelial dysfunction. We hypothesized that administration of a thiazolidinedione (TZD) agent would improve exercise capacity in type 2 diabetes. RESEARCH DESIGN AND METHODS: Twenty participants with uncomplicated type 2 diabetes were randomly assigned in a double-blind study to receive either 4 mg/day of rosiglitazone or matching placebo after baseline measurements to assess endothelial function (brachial artery diameter by brachial ultrasound), maximal oxygen consumption (VO(2max)), oxygen uptake (VO(2)) kinetics, and insulin sensitivity by hyperinsulinemic-euglycemic clamp. Measurements were reassessed after 4 months of treatment. RESULTS: Participant groups did not differ at baseline in any measure. Rosiglitazone-treated participants (n = 10) had significantly improved VO(2max) (19.8 +/- 5.3 ml . kg( 1) . min(-1) before rosiglitazone vs. 21.2 +/- 5.1 ml . kg(-1) . min(-1) after rosiglitazone, P < 0.01), insulin sensitivity, and endothelial function. A change in VO(2max) correlated with improved insulin sensitivity measured by clamp (r = 0.68, P < 0.05) and with improved brachial artery diameter (r = 0.70, P < 0.05). Placebo-treated participants (n = 10) showed no changes in VO(2max) (19.4 +/- 5.2 ml . kg(-1) . min(-1) before rosiglitazone vs. 18.1 +/- 5.3 ml . kg(-1) . min(-1) after rosiglitazone, NS) or brachial artery diameter. CONCLUSIONS: This is the first known report showing that a TZD improved exercise function in type 2 diabetes. Whether this is due to the observed improvements in insulin sensitivity and/or endothelial function or to another action of the TZD class requires further exploration. PMID- 16306549 TI - Modulation of food intake by glucose in patients with type 2 diabetes. AB - OBJECTIVE: A gain in body weight is a common adverse effect of glucose-lowering therapies in patients with type 2 diabetes, the mechanisms of which are not completely understood. Blood glucose is considered to play a crucial role in the regulation of food intake. On this background, we hypothesized that a short-term reduction of blood glucose concentration to normal values acutely increases food intake in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: To test this hypothesis, 12 patients with type 2 diabetes were examined twice, once during a euglycemic (5.0 mmol/l) clamp experiment and another time during a hyperglycemic (10.5 mmol/l) clamp. The experiments were performed in a single-blind fashion with the order of conditions balanced across patients. On both clamp conditions, insulin was infused at a constant rate of 2.5 mU/kg per min for 125 min. Simultaneously, a glucose solution was infused at a variable rate to achieve target glycemic levels. During the final 30 min of the clamps, the patients were allowed to eat as much as they liked from a standard breakfast buffet. RESULTS: Compared with the hyperglycemic condition, the patients ingested on average 25 +/ 10% more energy during euglycemia (645 +/- 75 vs. 483 +/- 37 kcal; P = 0.029). The increased energy intake during euglycemia was equally distributed across macronutrient components, i.e., during euglycemia the patients ate more carbohydrates (+27.1 +/- 11.4%; P = 0.037), fat (+22.5 +/- 10.0%; P = 0.046), and proteins (+25.2 +/- 11.2%; P = 0.046) than during hyperglycemia. Circulating levels of insulin, amylin, leptin, ghrelin, and glucagon-like peptide-1 did not differ between the euglycemic and hyperglycemia clamp, excluding a major contribution of these hormones to the difference in food intake. Summing up the glucose administered intravenously and the food ingested yielded a remarkably similar total energy influx in both conditions (794 +/- 64 vs. 790 +/- 53 kcal; P = 0.961). CONCLUSIONS: Together our data suggest that total energy supply to the organism is tightly regulated on a short-term basis independent of the route of influx. Alternatively, it can be hypothesized that euglycemia stimulated or that hyperglycemia suppressed food intake at the subsequent buffet meal in our type 2 diabetic patients. Regardless of these different interpretations, our data indicate an important regulatory role of glucose for food intake in type 2 diabetic patients that is of considerable clinical relevance. PMID- 16306550 TI - Serum interleukin-18 levels are associated with nephropathy and atherosclerosis in Japanese patients with type 2 diabetes. AB - OBJECTIVE: Interleukin (IL)-18 is a proinflammatory cytokine secreted from mononuclear cells. Serum concentration of IL-18 is a strong predictor of death in patients with cardiovascular diseases. Recent studies have shown that microinflammation is involved in the pathogenesis of diabetic nephropathy as well as of cardiovascular diseases. This study aimed to test the hypothesis that the serum level of IL-18 is a common predictor of nephropathy and atherosclerosis in patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: Eighty-two Japanese patients with type 2 diabetes and 55 age- and sex-matched healthy control subjects were enrolled. Patients with renal dysfunction (creatinine clearance <1 ml/s) were excluded. We assessed clinical parameters and measured serum and urinary IL-18 levels, serum IL-6 levels, carotid intima-media thickness (IMT), and brachial-ankle pulse wave velocity (baPWV) in all patients. Further, we evaluated changes of urinary albumin excretion rate (AER) after 6 months in 76 diabetic patients. RESULTS: Serum and urinary IL-18 levels were significantly elevated in patients with type 2 diabetes as compared with control subjects (serum IL-18 179 +/- 62 vs. 121 +/- 55 pg/ml, P < 0.001; urinary IL-18 97 +/- 159 vs. 47 +/- 54 pg/ml, P = 0.035). Univariate linear regression analysis showed significant positive correlations between serum IL-18 and AER (r [correlation coefficient] = 0.525, P < 0.001), HbA(1c) (r = 0.242, P = 0.029), high sensitivity C-reactive protein (hs-CRP) (r = 0.240, P = 0.031), and urinary beta 2 microglobulin (r = 0.235, P = 0.036). Serum IL-18 levels also correlated positively with carotid IMT (r = 0.225, P = 0.042) and baPWV (r = 0.232, P = 0.040). We also found a significant correlation between urinary IL-18 and AER (r = 0.309, P = 0.005). Multivariate linear regression analysis showed that AER (standard correlation coefficients [B] = 0.405, P < 0.001) and hs-CRP (B = 0.207, P = 0.033) were independently associated with serum IL-18 levels. AER was also independently associated with urinary IL-18 levels (B = 0.295, P = 0.005). Moreover, serum and urinary IL-18 levels correlated positively with AER after 6 months (r = 0.489, P < 0.001 and r = 0.320, P = 0.005) and changes in AER during the follow-up period (r = 0.268, P = 0.018 and r = 0.234, P = 0.042). CONCLUSIONS: Serum levels of IL-18 might be a predictor of progression of diabetic nephropathy as well as cardiovascular diseases. PMID- 16306551 TI - The effect of monochromatic infrared energy on sensation in patients with diabetic peripheral neuropathy: a double-blind, placebo-controlled study. AB - OBJECTIVE: The purpose of this study was to determine the effect of monochromatic infrared energy (MIRE) on plantar sensation in subjects with diabetic peripheral neuropathy. RESEARCH DESIGN AND METHODS: In this randomized, double-blind, placebo-controlled study, 39 subjects with diabetic peripheral neuropathy completed the 8-week study. Subjects received 30 min of active or placebo MIRE three times a week for 4 weeks. Plantar sensation was tested with monofilaments at the beginning of the study (M1), following 4 weeks of treatment (M2), and after an additional 4 weeks of nontreatment (M3). The number of sites that could sense the 5.07 monofilament was totaled at M1, M2, and M3. Data were analyzed using a special repeated-measures statistic followed by a post hoc Tukey-Kramer test. RESULTS: The average number of sites that patients could sense the 5.07 monofilament increased for both the active and placebo groups. There were significant gains from M1 to M2 (P < 0.002), no significant gains from M2 to M3 (P = 0.234), and significant gains from M1 to M3 (P < 0.002) for both the active and placebo groups. There were no significant differences between active and placebo groups at any measurement. CONCLUSIONS: Thirty minutes of active MIRE applied 3 days per week for 4 weeks was no more effective than placebo MIRE in increasing sensation in subjects with diabetic peripheral neuropathy. Clinicians should be aware that MIRE may not be an effective modality for improving sensory impairments in patients with diabetic neuropathy. PMID- 16306552 TI - Type 2 diabetes as a "coronary heart disease equivalent": an 18-year prospective population-based study in Finnish subjects. AB - OBJECTIVE: The purpose of this study was to investigate the hypothesis that coronary heart disease (CHD) mortality in diabetic subjects without prior evidence of CHD is equal to that in nondiabetic subjects with prior myocardial infarction or any prior evidence of CHD. RESEARCH DESIGN AND METHODS: During an 18-year follow-up total, cardiovascular disease (CVD) and CHD deaths were registered in a Finnish population-based study of 1,373 nondiabetic and 1,059 diabetic subjects. RESULTS: Adjusted multivariate Cox hazard models indicated that diabetic subjects without prior myocardial infarction, compared with nondiabetic subjects with prior myocardial infarction, had a hazard ratio (HR) of 0.9 (95% CI 0.6-1.5) for the risk of CHD death. The corresponding HR was 0.9 (0.5 1.4) in men and 1.9 (0.6 -6.1) in women. Diabetic subjects without any prior evidence of CHD (myocardial infarction or ischemic electrocardiogram [ECG] changes or angina pectoris), compared with nondiabetic subjects with prior evidence of CHD, had an HR of 1.9 (1.4-2.6) for CHD death (men 1.5 [1.0-2.2]; women 3.5 [1.8-6.8]). The results for CVD and total mortality were quite similar to those for CHD mortality. CONCLUSIONS: Diabetes without prior myocardial infarction and prior myocardial infarction without diabetes indicate similar risk for CHD death in men and women. However, diabetes without any prior evidence of CHD (myocardial infarction or angina pectoris or ischemic ECG changes) indicates a higher risk than prior evidence of CHD in nondiabetic subjects, especially in women. PMID- 16306553 TI - "Pressure gradient" as an indicator of plantar skin injury. AB - OBJECTIVE: Peak plantar pressures (PPPs) have been studied extensively as a contributing factor to skin breakdown, especially in the forefoot where most plantar neuropathic ulcers occur. The purposes of this article were to 1) describe an additional pressure variable, the peak pressure gradient (PPG), 2) determine whether the PPG is higher in the forefoot than in the rearfoot (even when compared with PPP), and 3) determine the correlation between the PPG and PPP at the forefoot and rearfoot in subjects with diabetes, peripheral neuropathy, and a history of plantar ulcer. RESEARCH DESIGN AND METHODS: Twenty subjects (12 male and 8 female) with diabetes, peripheral neuropathy, and a mean +/- SD age of 57 +/- 9 years participated. Plantar pressures were collected during walking in footwear. The PPP and the PPG (defined as the spatial change in plantar pressure across adjacent sites of the foot surface around the PPP) were determined for the forefoot and rearfoot, and the forefoot-to-rearfoot ratios for each variable were calculated. RESULTS: The mean PPG was 143% higher in the forefoot than in the rearfoot, whereas the mean PPP was only 36% higher in the forefoot than in the rearfoot (P < 0.0001). The PPG forefoot-to-rearfoot ratio (2.84 +/- 1.36) was nearly two times greater than the PPP forefoot-to-rearfoot ratio (1.48 +/- 0.58) (P < 0.0001). The correlation between PPP and PPG was r = 0.59 at the forefoot and r = 0.75 at the rearfoot. CONCLUSIONS: The PPG was substantially higher in the forefoot than in the rearfoot even when compared with the PPP. The PPG appears to be providing additional information about the stresses experienced by the soft tissues of the foot, especially in the forefoot. The PPG may be a useful indicator of skin trauma because spatial changes in high plantar pressures may identify high stress concentrations within the soft tissue. PMID- 16306554 TI - Hepatic enzymes, the metabolic syndrome, and the risk of type 2 diabetes in older men. AB - OBJECTIVE: We have examined the relationship between hepatic enzymes, the metabolic syndrome, insulin resistance, and type 2 diabetes and assessed the potential of hepatic enzyme measurements in determining diabetes risk. RESEARCH DESIGN AND METHODS: We conducted a prospective study of 3,500 nondiabetic men aged 60-79 years who were followed-up for a mean period of 5 years and in whom there were 100 incident type 2 diabetes cases. RESULTS: In cross-sectional analyses, alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) were strongly associated with obesity, insulin resistance, and the metabolic syndrome. Prospectively, the risk of type 2 diabetes significantly increased with increasing levels of ALT and GGT even after adjustment for confounders including BMI (top versus bottom quarter ALT: relative risk 2.72 [95% CI 1.47-5.02]; GGT: 3.68 [1.68-8.04]). Additional adjustment for insulin resistance attenuated the effects, but the relationships with ALT and GGT remained significant (1.91 [1.01 3.60] and 2.69 [1.21-5.97], respectively). Further adjustment for inflammatory markers (C-reactive protein) made minor differences. Among high-risk subjects (obese men or those with the metabolic syndrome), elevated GGT and ALT enhanced the prediction of diabetes risk. CONCLUSIONS: Elevated levels of ALT and GGT within the normal range are independent predictors of type 2 diabetes in older men and are useful additional measures in identifying those at high risk of diabetes. PMID- 16306555 TI - Anthropometry, glucose tolerance, and insulin concentrations in Indian children: relationships to maternal glucose and insulin concentrations during pregnancy. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that the environment experienced by fetuses of mothers with gestational diabetes mellitus (GDM) and mothers with higher glucose concentrations that are in the normal range causes increased adiposity and altered glucose/insulin metabolism in childhood. RESEARCH DESIGN AND METHODS: Children (n = 630) whose mothers were tested for glucose tolerance during pregnancy had detailed anthropometry performed at birth and annually thereafter. At 5 years, plasma glucose and insulin concentrations were measured in the children (2-h oral glucose tolerance test) and their fathers (fasting samples only). RESULTS: Newborns of diabetic mothers (n = 41) were larger in all body measurements than control newborns (babies with nondiabetic parents). At 1 year, these differences had diminished and were not statistically significant. At 5 years, female offspring of diabetic mothers had larger subscapular and triceps skinfold thicknesses (P = 0.01) and higher 30- and 120 min insulin concentrations (P < 0.05) than control children. Offspring of diabetic fathers (n = 41) were lighter at birth than control children (P < 0.001); they showed no differences in anthropometry at 5 years. In control children, skinfold thickness and 30-min insulin concentrations were positively related to maternal insulin area under the curve, and skinfold thicknesses were related to paternal fasting insulin concentrations independently of the parents' skinfold thickness and socioeconomic status. CONCLUSIONS: Maternal GDM is associated with adiposity and higher glucose and insulin concentrations in female offspring at 5 years. The absence of similar associations in offspring of diabetic fathers suggests a programming effect in the diabetic intrauterine environment. More research is needed to determine whether higher maternal glucose concentrations in the nondiabetic range have similar effects. PMID- 16306556 TI - Dietary calcium, vitamin D, and the prevalence of metabolic syndrome in middle aged and older U.S. women. AB - OBJECTIVE: To examine whether and to what extent intakes of calcium and vitamin D are related to the metabolic syndrome in middle-aged or older women. RESEARCH DESIGN AND METHODS: We analyzed data from 10,066 women aged > or =45 years participating in the Women's Health Study who were free of cardiovascular disease, cancer, or diabetes and who never used postmenopausal hormones. We used multiple logistic regression models to estimate multivariable odds ratios (ORs) and 95% CIs comparing different dietary intake levels of calcium and vitamin D. RESULTS: In age- and calorie-adjusted analyses, higher intakes of total, dietary, and supplemental calcium were significantly and inversely associated with the prevalence of metabolic syndrome. After further adjusting for smoking status, exercise, alcohol intake, multivitamin use, and parental history of myocardial infarction before age 60 years, the ORs of having the metabolic syndrome for increasing quintiles of total calcium intake were 1.00 (reference), 0.82 (95% CI 0.70-0.97), 0.84 (0.71-0.99), 0.70 (0.59-0.83), and 0.64 (0.54-0.77) (P for trend <0.0001). This association was not appreciably altered by additional adjustment for other dietary factors or total vitamin D intake. In contrast, neither total (P for trend = 0.13) nor supplemental (P for trend = 0.45) vitamin D was significantly associated with metabolic syndrome. Dietary vitamin D was inversely associated with prevalence of metabolic syndrome but was not independent of total calcium intake. Similar strong relations between intakes of dairy products and metabolic syndrome were also observed. After adjustment for lifestyle and dietary factors, the multivariable ORs comparing highest with lowest intake categories were 0.66 (0.55-0.80) (P for trend <0.0001) for total dairy products and 0.85 (0.71-1.02) (P for trend = 0.05) for total milk intake. CONCLUSIONS: Our results indicate that intakes of calcium and dairy products may be associated with lower prevalence of the metabolic syndrome in middle-aged and older women. PMID- 16306557 TI - Alcohol consumption and risk of type 2 diabetes among older women. AB - OBJECTIVE: This study aimed to investigate the relation between alcohol consumption and type 2 diabetes among older women. RESEARCH DESIGN AND METHODS: Between 1993 and 1997, 16,330 women aged 49-70 years and free from diabetes were enrolled in one of the Dutch Prospect-EPIC (European Prospective Study Into Cancer and Nutrition) cohorts and followed for 6.2 years (range 0.1-10.1). At enrollment, women filled in questionnaires and blood samples were collected. RESULTS: During follow-up, 760 cases of type 2 diabetes were documented. A linear inverse association (P = 0.007) between alcohol consumption and type 2 diabetes risk was observed, adjusting for potential confounders. Compared with abstainers, the hazard ratio for type 2 diabetes was 0.86 (95% CI 0.66-1.12) for women consuming 5-30 g alcohol per week, 0.66 (0.48-0.91) for 30-70 g per week, 0.91 (0.67-1.24) for 70-140 g per week, 0.64 (0.44-0.93) for 140-210 g per week, and 0.69 (0.47-1.02) for >210 g alcohol per week. Beverage type did not influence this association. Lifetime alcohol consumption was associated with type 2 diabetes in a U-shaped fashion. CONCLUSIONS: Our findings support the evidence of a decreased risk of type 2 diabetes with moderate alcohol consumption and expand this to a population of older women. PMID- 16306558 TI - A low-glycemic load diet facilitates greater weight loss in overweight adults with high insulin secretion but not in overweight adults with low insulin secretion in the CALERIE Trial. PMID- 16306559 TI - Lack of effect of guideline changes on LDL cholesterol reporting and control for diabetes visits in the U.S., 1995-2004. PMID- 16306560 TI - Diabetes and the labor market: the community-wide economic cost in the Lower Rio Grande Valley. PMID- 16306561 TI - Hypoglycemia in type 2 diabetes: pathophysiology, frequency, and effects of different treatment modalities. PMID- 16306562 TI - Acute neuropathic joint disease: a medical emergency? PMID- 16306563 TI - Commentary on the results and clinical implications of the PROactive study. PMID- 16306564 TI - Clinical diabetic neuropathy. PMID- 16306565 TI - Why might thiazolidinediones increase exercise capacity in patients with type 2 diabetes? PMID- 16306566 TI - Do glycemic index, glycemic load, and fiber play a role in insulin sensitivity, disposition index, and type 2 diabetes? PMID- 16306567 TI - Efficacy of pitavastatin, a new HMG-CoA reductase inhibitor, on lipid and glucose metabolism in patients with type 2 diabetes. PMID- 16306568 TI - Type III allergy to insulin detemir. PMID- 16306569 TI - Twelve-hour glycemic profiles with meals of high, medium, or low glycemic load. PMID- 16306570 TI - A case of lipoatrophy with insulin glargine: long-acting insulin analogs are not exempt from this complication. PMID- 16306571 TI - Induction of innate immune response genes by Sin Nombre hantavirus does not require viral replication. AB - Maladaptive immune responses are considered to be important factors in the pathogenesis of the two diseases caused by hantaviruses, hemorrhagic fever with renal syndrome and hantavirus cardiopulmonary syndrome (HCPS). While the intensity of adaptive antiviral T-cell responses seems to correlate with the severity of HCPS, there is increasing evidence that innate antiviral responses by endothelial cells, the native targets for hantavirus infection in vivo, are induced within hours of exposure to infectious hantaviruses. To investigate early events in the innate response to Sin Nombre virus (SNV), the principal etiologic agent of HCPS in North America, we treated human endothelial cells with live virus, or virus subjected to inactivation by UV irradiation at minimal doses required to inhibit replication, and assayed host expression of interferon stimulated genes (ISG) by microarray and reverse transcription-PCR. We show herein that a variety of ISG are induced between 4 and 24 h after exposure to both live and killed virus. The levels of such induction at early time points (before 24 h) were generally higher in cells treated with SNV particles that had been killed by exposure to UV irradiation. Additionally, SNV exposed to increasing doses of UV irradiation induced ISG better than live virus despite increased disruption of viral RNA integrity. However, SNV replication was required for continued ISG overexpression by 3 days posttreatment. These results suggest that hantavirus particles may themselves be capable of early induction of ISG and that ongoing production of viral particles during infection could contribute to the pathogenic process. PMID- 16306572 TI - A point mutation within the replicase gene differentially affects coronavirus genome versus minigenome replication. AB - During the construction of the transmissible gastroenteritis virus (TGEV) full length cDNA clone, a point mutation at position 637 that was present in the defective minigenome DI-C was maintained as a genetic marker. Sequence analysis of the recovered viruses showed a reversion at this position to the original virus sequence. The effect of point mutations at nucleotide 637 was analyzed by reverse genetics using a TGEV full-length cDNA clone and cDNAs from TGEV-derived minigenomes. The replacement of nucleotide 637 of TGEV genome by a T, as in the DI-C sequence, or an A severely affected virus recovery from the cDNA, yielding mutant viruses with low titers and small plaques compared to those of the wild type. In contrast, T or A at position 637 was required for minigenome rescue in trans by the helper virus. No relationship between these observations and RNA secondary-structure predictions was found, indicating that mutations at nucleotide 637 most likely had an effect at the protein level. Nucleotide 637 occupies the second codon position at amino acid 108 of the pp1a polyprotein. This position is predicted to map in the N-terminal polyprotein papain-like proteinase (PLP-1) cleavage site at the p9/p87 junction. Replacement of G-637 by A, which causes a drastic amino acid change (Gly to Asp) at position 108, affected PLP-1-mediated cleavage in vitro. A correlation was found between predicted cleaving and noncleaving mutations and efficient virus rescue from cDNA and minigenome amplification, respectively. PMID- 16306573 TI - Kaposi's sarcoma-associated herpesvirus induction of AP-1 and interleukin 6 during primary infection mediated by multiple mitogen-activated protein kinase pathways. AB - Kaposi's sarcoma is an angioproliferative disseminated tumor of endothelial cells linked to infection with Kaposi's sarcoma-associated herpesvirus (KSHV). AP-1 transcription factors are involved in diverse biological processes, including infection and replication of viruses, cell growth, oncogenesis, angiogenesis, and invasion of cancer cells. Here we show that KSHV activates AP-1 during primary infection. The activation of AP-1 at the early stage of KSHV infection is mainly mediated by virus entry events. Concurrently, KSHV infection strongly activates MEK, JNK, and to a lesser extent, p38 mitogen-activated protein kinase (MAPK) pathways. Specific inhibitors or dominant negative constructs of MEK and JNK completely abolish AP-1 activation by KSHV, while those of p38 reduce it by half. Furthermore, individual MAPK pathways differentially regulate KSHV activation of AP-1 components. KSHV activation of AP-1 leads to the transcriptional induction of interleukin 6 (IL-6), which is inhibited by inhibitors or dominant negative constructs of MAPK pathways. Together, these results demonstrate that KSHV induces AP-1 and IL-6 during primary infection by modulating multiple MAPK pathways. Because of the diverse roles of IL-6, AP-1, and MAPK pathways in viral infection and tumor induction and promotion, these results have important implications in the pathogenesis of KSHV-induced malignancies. PMID- 16306574 TI - Simultaneous mutations in CA and Vif of Maedi-Visna virus cause attenuated replication in macrophages and reduced infectivity in vivo. AB - Maedi-visna virus (MVV) is a lentivirus of sheep sharing several key features with the primate lentiviruses. The virus causes slowly progressive diseases, mainly in the lungs and the central nervous system of sheep. Here, we investigate the molecular basis for the differential growth phenotypes of two MVV isolates. One of the isolates, KV1772, replicates well in a number of cell lines and is highly pathogenic in sheep. The second isolate, KS1, no longer grows on macrophages or causes disease. The two virus isolates differ by 129 nucleotide substitutions and two deletions of 3 and 15 nucleotides in the env gene. To determine the molecular nature of the lesions responsible for the restrictive growth phenotype, chimeric viruses were constructed and used to map the phenotype. An L120R mutation in the CA domain, together with a P205S mutation in Vif (but neither alone), could fully convert KV1772 to the restrictive growth phenotype. These results suggest a functional interaction between CA and Vif in MVV replication, a property that may relate to the innate antiretroviral defense mechanisms in sheep. PMID- 16306575 TI - A vesicular stomatitis virus recombinant expressing granulocyte-macrophage colony stimulating factor induces enhanced T-cell responses and is highly attenuated for replication in animals. AB - Live attenuated vectors based on recombinant vesicular stomatitis viruses (rVSVs) expressing foreign antigens are highly effective vaccines in animal models. In this study, we report that an rVSV (VSV-GMCSF1) expressing high levels of murine granulocyte-macrophage colony-stimulating factor (GM-CSF) from the first position in the viral genome is highly attenuated in terms of viral dissemination and pathogenesis after intranasal delivery to mice. However, this highly attenuated virus generated antibody and T-cell responses equivalent to those induced by a control virus expressing enhanced green fluorescent protein (EGFP) from the first position (VSV-EGFP1). The better containment and clearance of VSV-GMCSF1 may be due to enhanced recruitment of macrophages to the site of infection but is not explained by a greater induction of interferons. The primary CD8 T-cell and neutralizing antibody responses to VSV-GMCSF1 were equivalent to those generated by VSV-EGFP1, while the CD8 T-cell memory and recall responses to the vector were enhanced in mice infected with VSV-GMCSF1. It is likely that the GM-CSF produced by immunization with this virus results in an enhanced recruitment of antigen presenting cells, leading to better acute and long-term T-cell responses. This recruitment appears to cancel out any negative effect of viral attenuation on immunogenicity. PMID- 16306576 TI - Luxury at a cost? Recombinant mouse hepatitis viruses expressing the accessory hemagglutinin esterase protein display reduced fitness in vitro. AB - Group 2 coronaviruses encode an accessory envelope glycoprotein species, the hemagglutinin esterase (HE), which possesses sialate-O-acetylesterase activity and which, presumably, promotes virus spread and entry in vivo by facilitating reversible virion attachment to O-acetylated sialic acids. While HE may provide a strong selective advantage during natural infection, many laboratory strains of mouse hepatitis virus (MHV) fail to produce the protein. Apparently, their HE genes were inactivated during cell culture adaptation. For this report, we have studied the molecular basis of this phenomenon. By using targeted RNA recombination, we generated isogenic recombinant MHVs which differ exclusively in their expression of HE and produce either the wild-type protein (HE+), an enzymatically inactive HE protein (HE0), or no HE at all. HE expression or the lack thereof did not lead to gross differences in in vitro growth properties. Yet the expression of HE was rapidly lost during serial cell culture passaging. Competition experiments with mixed infections revealed that this was not due to the enzymatic activity: MHVs expressing HE+ or HE0 propagated with equal efficiencies. During the propagation of recombinant MHV-HE+, two types of spontaneous mutants accumulated. One produced an anchorless HE, while the other had a Gly-to-Trp substitution at the predicted C-terminal residue of the HE signal peptide. Neither mutant incorporated HE into virion particles, suggesting that wild-type HE reduces the in vitro propagation efficiency, either at the assembly stage or at a postassembly level. Our findings demonstrate that the expression of "luxury" proteins may come at a fitness penalty. Apparently, under natural conditions the costs of maintaining HE are outweighed by the benefits. PMID- 16306577 TI - Expression of hemagglutinin esterase protein from recombinant mouse hepatitis virus enhances neurovirulence. AB - Murine hepatitis virus (MHV) infection provides a model system for the study of hepatitis, acute encephalitis, and chronic demyelinating disease. The spike glycoprotein, S, which mediates receptor binding and membrane fusion, plays a critical role in MHV pathogenesis. However, viral proteins other than S also contribute to pathogenicity. The JHM strain of MHV is highly neurovirulent and expresses a second spike glycoprotein, the hemagglutinin esterase (HE), which is not produced by MHV-A59, a hepatotropic but only mildly neurovirulent strain. To investigate a possible role for HE in MHV-induced neurovirulence, isogenic recombinant MHV-A59 viruses were generated that produced either (i) the wild-type protein, (ii) an enzymatically inactive HE protein, or (iii) no HE at all (A. Lissenberg, M. M. Vrolijk, A. L. W. van Vliet, M. A. Langereis, J. D. F. de Groot Mijnes, P. J. M. Rottier, and R. J. de Groot, J. Virol. 79:15054-15063, 2005 [accompanying paper]). A second, mirror set of recombinant viruses was constructed in which, in addition, the MHV-A59 S gene had been replaced with that from MHV-JHM. The expression of HE in combination with A59 S did not affect the tropism, pathogenicity, or spread of the virus in vivo. However, in combination with JHM S, the expression of HE, regardless of whether it retained esterase activity or not, resulted in increased viral spread within the central nervous system and in increased neurovirulence. Our findings suggest that the properties of S receptor utilization and/or fusogenicity mainly determine organ and host cell tropism but that HE enhances the efficiency of infection and promotes viral dissemination, at least in some tissues, presumably by serving as a second receptor-binding protein. PMID- 16306578 TI - Ubiquitination of the prototype foamy virus envelope glycoprotein leader peptide regulates subviral particle release. AB - Foamy virus (FV) particle egress is unique among retroviruses because of its essential requirement for Gag and Env coexpression for budding and particle release. The FV glycoprotein undergoes a highly unusual biosynthesis resulting in the generation of three particle-associated, mature subunits, leader peptide (LP), surface (SU), and transmembrane (TM), derived from a precursor protein by posttranslational proteolysis mediated by furin or furinlike proteases. Previously at least three LP products of different molecular weights were detected in purified FV particles. Here we demonstrate that the higher-molecular weight forms gp28LP and gp38LP are ubiquitinated variants of the major gp18LP cleavage product, which has a type II membrane topology. Furthermore, we show that all five lysine residues located within the N-terminal 60-amino-acid cytoplasmic domain of gp18LP can potentially be ubiquitinated, however, there seems to be a preference for using the first three. Inactivation of ubiquitination sites individually resulted in no obvious phenotype. However, simultaneous inactivation of the first three or all five ubiquitination sites in gp18LP led to a massive increase in subviral particles released by these mutant glycoproteins that were readily detectable by electron microscopy analysis upon expression of the ubiquitination-deficient glycoprotein by itself or in a proviral context. Surprisingly, only the quintuple ubiquitination mutant showed a two- to threefold increase in single-cycle infectivity assays, whereas all other mutants displayed infectivities similar to that of the wild type. Taken together, these data suggest that the balance between viral and subviral particle release of FVs is regulated by ubiquitination of the glycoprotein LP. PMID- 16306579 TI - Vaccinia virus nicking-joining enzyme is encoded by K4L (VACWR035). AB - Vaccinia virus encodes an enzyme with DNA modifying activity that cleaves and inefficiently cross-links cruciformic DNA. This enzyme is contained within the virion, expressed at late times postinfection, and processes DNA in an energy independent, Mg2+ ion-independent manner. Viral nuclease activity was measured in extracts from cells infected with well-defined viral mutants. Since some viral extracts lacked nuclease activity, the gene encoding the activity was postulated to be one of the open reading frames absent in the viruses lacking activity. Inducible expression of each candidate open reading frame revealed that only the gene VACWR035, or K4L, was required for nuclease activity. A recombinant virus missing only the open reading frame for K4L lacked nuclease activity. Extracts from a recombinant virus expressing K4L linked to a FLAG polypeptide were able to cleave and cross-link cruciformic DNA. There were no significant differences between the virus lacking K4L and wild-type vaccinia virus WR with respect to infectivity, growth characteristics, or processing of viral replicative intermediate DNA, including both telomeric and cross-linked forms. Purification of the K4L FLAG polypeptide expressed in bacteria yielded protein containing nicking-joining activity, implying that K4L is the only vaccinia virus protein required for the nicking-joining enzymatic activity. PMID- 16306580 TI - Simian immunodeficiency virus variants that differ in pathogenicity differ in fitness under rapid cell turnover conditions. AB - Simian immunodeficiency virus (SIV) has been shown to progress through a number of changes that lead to the emergence of pathogenic viral variants in macaques initially infected with a mildly cytopathic variant, SIVMneCL8. One of these late stage isolates, SIVMne170, replicates to high levels in vivo and causes a rapid disease course when reintroduced into naive macaques, resulting in a viral set point up to 3,000-fold higher than the set point of the parental virus, SIVMneCL8. However, in cell culture both viruses replicate with similar kinetics. One major difference between in vivo and in vitro cultures is the life span of the infected cells. Here, we manipulated the life span of infected cells in vitro, and we show that the fitness of SIVMne170 in cultures with a limited cell life span dramatically increased compared to its fitness in cultures with a nonlimited life span of cells. The increase in fitness was at least partially due to the fact that the rapid turnover system eliminates the negative influence of the cytopathic effects associated with replication of SIVMne170. Because the relative fitness of SIVMneCL8 and SIVMne170 observed in the rapid turnover system more accurately reflects their fitness in vivo, the system represents an improved approach to comparing relative fitness of viruses. PMID- 16306581 TI - Transcriptional analysis of latent and inducible Kaposi's sarcoma-associated herpesvirus transcripts in the K4 to K7 region. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is a gamma-2 herpesvirus with a genome containing a long unique coding region (LUR) flanked by GC-rich terminal repeat sequences. The LUR encodes approximately 90 annotated open reading frames (ORFs) with complex patterns of gene expression during viral latency, reactivation, and de novo infection. To identify unannotated KSHV genes, we examined the region between 21,500 and 30,000 bp of the KSHV LUR, representing approximately 8.5 kb of sequence. This region encodes seven known single-exon ORFs (K4, K4.1, K4.2, K5, K6, K7, and PAN), but previous computer analyses have failed to identify additional likely genes in the remaining 5.2 kb. We identified four novel transcripts using Northern blotting, phage library screening, and 5' rapid amplification of cDNA ends analysis in the region between ORFs K4.2 and K7. In vitro analysis of KSHV-infected primary effusion lymphoma cell lines in the presence of 12-O-tetradecanoylphorbol-13-acetate and phosphonoformic acid suggests that one latent transcript is coterminal with the previously annotated K3 gene encoding an ubiquitin-ligase known to downregulate major histocompatibility complex class I expression. This alternatively spliced transcript may contribute to KSHV adaptive immune evasion during latent infection. Other transcripts are inducible, including a 6.1-kb transcript that is the largest transcript found in the KSHV genome to date. PMID- 16306582 TI - Humoral and cellular immune response to RNA immunization with flavivirus replicons derived from tick-borne encephalitis virus. AB - A new vaccination principle against flaviviruses, based on a tick-borne encephalitis virus (TBEV) self-replicating noninfectious RNA vaccine that produces subviral particles, has recently been introduced (R. M. Kofler, J. H. Aberle, S. W. Aberle, S. L. Allison, F. X. Heinz, and C. W. Mandl, Proc. Natl. Acad. Sci. USA 7:1951-1956, 2004). In this study, we evaluated the potential of the self-replicating RNA vaccine in mice in comparison to those of live, attenuated vaccines and a formalin-inactivated whole-virus vaccine (ImmunInject). For this purpose, mice were immunized using gene gun-mediated application of the RNA vaccine and tested for CD8+ T-cell responses, long-term duration, neutralizing capacity, and isotype profile of specific antibodies and protection against lethal virus challenge. We demonstrate that the self-replicating RNA vaccine induced a broad-based, humoral and cellular (Th1 and CD8+ T-cell response) immune response comparable to that induced by live vaccines and that it protected mice from challenge. Even a single immunization with 1 microg of the replicon induced a long-lasting antibody response, characterized by high neutralizing antibody titers, which were sustained for at least 1 year. Nevertheless, it was possible to boost this response further by a second injection with the RNA vaccine, even in the presence of a concomitant CD8+ T-cell response. In this way it was possible to induce a balanced humoral and cellular immune response, similar to infection-induced immunity but without the safety hazards of infectious agents. The results also demonstrate the value of TBEV replicon RNA for inducing protective long-lasting antiviral responses. PMID- 16306583 TI - Chimeric recombinant human metapneumoviruses with the nucleoprotein or phosphoprotein open reading frame replaced by that of avian metapneumovirus exhibit improved growth in vitro and attenuation in vivo. AB - Chimeric versions of recombinant human metapneumovirus (HMPV) were generated by replacing the nucleoprotein (N) or phosphoprotein (P) open reading frame with its counterpart from the closely related avian metapneumovirus (AMPV) subgroup C. In Vero cells, AMPV replicated to an approximately 100-fold-higher titer than HMPV. Surprisingly, the N and P chimeric viruses replicated to a peak titer that was 11 and 25-fold higher, respectively, than that of parental HMPV. The basis for this effect is not known but was not due to obvious changes in the efficiency of gene expression. AMPV and the N and P chimeras were evaluated for replication, immunogenicity, and protective efficacy in hamsters. AMPV was attenuated compared to HMPV in this mammalian host on day 5 postinfection, but not on day 3, and only in the nasal turbinates. In contrast, the N and P chimeras were reduced approximately 100-fold in both the upper and lower respiratory tract on day 3 postinfection, although there was little difference by day 5. The N and P chimeras induced a high level of neutralizing serum antibodies and protective efficacy against HMPV; AMPV was only weakly immunogenic and protective against HMPV challenge, reflecting antigenic differences. In African green monkeys immunized intranasally and intratracheally, the mean peak titer of the P chimera was reduced 100- and 1,000-fold in the upper and lower respiratory tracts, whereas the N chimera was reduced only 10-fold in the lower respiratory tract. Both chimeras were comparable to wild-type HMPV in immunogenicity and protective efficacy. Thus, the P chimera is a promising live HMPV vaccine candidate that paradoxically combines improved growth in vitro with attenuation in vivo. PMID- 16306584 TI - Clade replacements in dengue virus serotypes 1 and 3 are associated with changing serotype prevalence. AB - The evolution of dengue virus (DENV) is characterized by phylogenetic trees that have a strong temporal structure punctuated by dramatic changes in clade frequency. To determine the cause of these large-scale phylogenetic patterns, we examined the evolutionary history of DENV serotype 1 (DENV-1) and DENV-3 in Thailand, where gene sequence and epidemiological data are relatively abundant over a 30-year period. We found evidence for the turnover of viral clades in both serotypes, most notably in DENV-1, where a major clade replacement event took place in genotype I during the mid-1990s. Further, when this clade replacement event was placed in the context of changes in serotype prevalence in Thailand, a striking pattern emerged; an increase in DENV-1 clade diversity was associated with an increase in the abundance of this serotype and a concomitant decrease in DENV-4 prevalence, while clade replacement was associated with a decline in DENV 1 prevalence and a rise of DENV-4. We postulate that intraserotypic genetic diversification proceeds at times of relative serotype abundance and that replacement events can result from differential susceptibility to cross-reactive immune responses. PMID- 16306585 TI - Effect of bottlenecking on evolution of the nonstructural protein 3 gene of hepatitis C virus during sexually transmitted acute resolving infection. AB - Sexual partners of patients infected with the hepatitis C virus (HCV) often have detectable HCV-specific T-cell responses in the absence of seroconversion, suggesting unapparent, spontaneously resolving infection. To determine whether differences in the evolutionary potential of bottlenecked inoculum may explain the low rate of HCV persistence after sexual exposure, we have investigated changes in the entire HCV nonstructural 3 (NS3) gene over time in a chronic carrier and compared his viral quasispecies with that of the acute-phase isolate of his sexual partner, who developed acute resolving hepatitis C. The overall rate of accumulation of mutations, estimated by regression analysis of six consecutive consensus NS3 sequences over 8 years, was 1.5 x 10(-3) mutations per site per year, with small intersample fluctuations related to changes in environmental conditions. Comparison of quasispecies parameters in one isolate of the chronic carrier with those of the acute-phase isolate of the infected partner revealed a higher heterogeneity and lower proportion of nonsynonymous mutations in the former. All NS3 sequences from the acute-phase isolate clustered with a single sequence from the chronic isolate, despite complete HLA mismatch between the patients, suggesting bottlenecking during transmission. The low risk of viral persistence after sexual exposure to HCV may be related to the selection of a limited number of viral particles carrying a particular combination of mutations which may further limit the potential of a relatively homogeneous quasispecies to rapidly diversify and overcome the immune response of the exposed host. PMID- 16306586 TI - Blockade of neutrophil elastase attenuates severe liver injury in hepatitis B transgenic mice. AB - Serine proteinases produced by polymorphonuclear neutrophils play important roles in neutrophil-mediated tissue injury at inflammatory sites. Although neutrophil recruitment to the liver has been shown to be involved in the exacerbation of liver inflammation, the function of neutrophil elastase (NE) in liver injury remains unclear. Here, we found that administration of an NE inhibitor (NEI) reduced serum alanine aminotransferase (sALT) activity and inflammatory cell infiltration into the liver from 8 to 24 h after injection of antigen-specific cytotoxic T lymphocytes (CTLs) into hepatitis B virus transgenic mice. Furthermore, the NEI treatment reduced the expressions of inflammatory cytokines and chemokines in the liver and tumor necrosis factor alpha production by macrophages. In addition, the NEI treatment suppressed the mRNA expressions of CC chemokine ligand 3 (CCL-3), CCL-4, and macrophage inflammatory protein 2 (MIP-2) in neutrophils in the liver at 8 h after the CTL injection. In support of these results, we confirmed that administration of anti-CCL-3, anti-CCL-4, and anti-MIP 2 monoclonal antibodies suppressed sALT activity and leukocyte migration into the liver. In conclusion, the present results suggest that NE contributes to the early step of the inflammatory cascade in acute viral hepatitis and that NEIs may have potential as therapeutic drugs against acute severe viral hepatitis. PMID- 16306587 TI - Transcription program of red sea bream iridovirus as revealed by DNA microarrays. AB - Red sea bream iridovirus (RSIV) has been identified as the causative agent of a serious disease in red sea bream and at least 30 other marine fish species. We developed a viral DNA microarray containing 92 putative open reading frames of RSIV to monitor the viral gene transcription program over the time course of an in vitro infection and to classify RSIV transcripts into temporal kinetic expression classes. The microarray analysis showed that viral genes commenced expression as early as 3 h postinfection (p.i.) and this was followed by a rapid escalation of gene expression from 8 h p.i. onwards. Based on the expression of some enzymes associated with viral DNA replication, the DNA replication of RSIV appeared to begin at around 8 h p.i. in infected cells in vitro. Using a de novo protein synthesis inhibitor (cycloheximide) and a viral DNA replication inhibitor (phosphonoacetic acid), 87 RSIV transcripts could be classified into three temporal kinetic classes: nine immediate-early (IE), 40 early (E), and 38 late (L) transcripts. The gene expression of RSIV occurred in a temporal kinetic cascade with three stages (IE, E, and L). Although the three classes of transcripts were distributed throughout the RSIV genome, E transcripts appeared to cluster in at least six discrete regions and L transcripts appeared to originate from seven discrete regions. The microarray data were statistically confirmed by using a t test, and were also clustered into groups based on similarity in the gene expression patterns by using a cluster program. PMID- 16306588 TI - Rotavirus glycoprotein NSP4 is a modulator of viral transcription in the infected cell. AB - The outer shell of the rotavirus triple-layered virion is lost during cell entry, yielding a double-layered particle (DLP) that directs synthesis of viral plus strand RNAs. The plus-strand RNAs act as templates for synthesis of the segmented double-stranded RNA (dsRNA) genome in viral inclusion bodies (viroplasms). The viral endoplasmic reticulum (ER)-resident glycoprotein NSP4 recruits progeny DLPs formed in viroplasms to the ER, where the particles are converted to triple layered particles (TLPs) via budding. In this study, we have used short interfering RNAs to probe the role of NSP4 in the viral life cycle. Our analysis showed that knockdown of NSP4 expression had no marked effect on the expression of other viral proteins or on the replication of the dsRNA genome segments. However, NSP4 loss of function suppressed viroplasm maturation and caused a maldistribution of nonstructural and structural proteins that normally accumulate in viroplasms. NSP4 loss of function also inhibited formation of packaged virus particles, instead inducing the accumulation of empty particles. Most significant was the observation that NSP4 knockdown led to dramatically increased levels of viral transcription late in the infection cycle. These findings point to a multifaceted role for NSP4 in virus replication, including influencing the development of viroplasms, linking genome packaging with particle assembly, and acting as a modulator of viral transcription. By recruiting transcriptionally active or potentially active DLPs to the ER for conversion to quiescent TLPs, NSP4 acts as a feedback inhibitor down-regulating viral transcription when adequate levels of plus-strand RNAs are available to allow for productive infection. PMID- 16306589 TI - Restriction of feline immunodeficiency virus by Ref1, Lv1, and primate TRIM5alpha proteins. AB - The Ref1 and Lv1 postentry restrictions in human and monkey cells have been analyzed for lentiviruses in the primate and ungulate groups, but no data exist for the third (feline) group. We compared feline immunodeficiency virus (FIV) to other restricted (human immunodeficiency virus type 1 [HIV-1], equine infectious anemia virus [EIAV]) and unrestricted (NB-tropic murine leukemia virus [NB-MLV]) retroviruses across wide ranges of viral inputs in cells from multiple primate and nonprimate species. We also characterized restrictions conferred to permissive feline and canine cells engineered to express rhesus and human TRIM5alpha proteins and performed RNA interference (RNAi) against endogenous TRIM5alpha. We find that expression of rhesus or human TRIM5alpha proteins in feline cells restricts FIV, impairing pseudotyped vector transduction and viral replication, but rhesus TRIM5alpha is more restricting than human TRIM5alpha. Notably, however, canine cells did not support restriction by human TRIM5alpha and supported minimal restriction by rhesus TRIM5alpha, suggesting that these proteins may not function autonomously or that a canine factor interferes. Stable RNAi knockdown of endogenous rhesus TRIM5alpha resulted in marked increases in FIV and HIV-1 infectivities while having no effect on NB-MLV. A panel of nonprimate cell lines varied widely in susceptibility to lentiviral vector transduction, but normalized FIV and HIV-1 vectors varied concordantly. In contrast, in human and monkey cells, relative restriction of FIV compared to HIV 1 varied from none to substantial, with the greatest relative infectivity deficit for FIV vectors observed in human T-cell lines. Endogenous and introduced TRIM5alpha restrictions of FIV could be titrated by coinfections with FIV, HIV-1, or EIAV virus-like particles. Arsenic trioxide had complex and TRIM5alpha independent enhancing effects on lentiviral but not NB-MLV infection. Implications for human gene therapy are discussed. PMID- 16306590 TI - The papain-like protease of severe acute respiratory syndrome coronavirus has deubiquitinating activity. AB - Replication of the genomic RNA of severe acute respiratory syndrome coronavirus (SARS-CoV) is mediated by replicase polyproteins that are processed by two viral proteases, papain-like protease (PLpro) and 3C-like protease (3CLpro). Previously, we showed that SARS-CoV PLpro processes the replicase polyprotein at three conserved cleavage sites. Here, we report the identification and characterization of a 316-amino-acid catalytic core domain of PLpro that can efficiently cleave replicase substrates in trans-cleavage assays and peptide substrates in fluorescent resonance energy transfer-based protease assays. We performed bioinformatics analysis on 16 papain-like protease domains from nine different coronaviruses and identified a putative catalytic triad (Cys1651 His1812-Asp1826) and zinc-binding site. Mutagenesis studies revealed that Asp1826 and the four cysteine residues involved in zinc binding are essential for SARS CoV PLpro activity. Molecular modeling of SARS-CoV PLpro suggested that this catalytic core may also have deubiquitinating activity. We tested this hypothesis by measuring the deubiquitinating activity of PLpro by two independent assays. SARS CoV-PLpro hydrolyzed both diubiquitin and ubiquitin-7-amino-4-methylcoumarin (AMC) substrates, and hydrolysis of ubiquitin-AMC is approximately 180-fold more efficient than hydrolysis of a peptide substrate that mimics the PLpro replicase recognition sequence. To investigate the critical determinants recognized by PLpro, we performed site-directed mutagenesis on the P6 to P2' residues at each of the three PLpro cleavage sites. We found that PLpro recognizes the consensus cleavage sequence LXGG, which is also the consensus sequence recognized by cellular deubiquitinating enzymes. This similarity in the substrate recognition sites should be considered during the development of SARS-CoV PLpro inhibitors. PMID- 16306591 TI - The papain-like protease from the severe acute respiratory syndrome coronavirus is a deubiquitinating enzyme. AB - The severe acute respiratory syndrome coronavirus papain-like protease (SARS-CoV PLpro) is involved in the processing of the viral polyprotein and, thereby, contributes to the biogenesis of the virus replication complex. Structural bioinformatics has revealed a relationship for the SARS-CoV PLpro to herpesvirus associated ubiquitin-specific protease (HAUSP), a ubiquitin-specific protease, indicating potential deubiquitinating activity in addition to its function in polyprotein processing (T. Sulea, H. A. Lindner, E. O. Purisima, and R. Menard, J. Virol. 79:4550-4551, 2005). In order to confirm this prediction, we overexpressed and purified SARS-CoV PLpro (amino acids [aa]1507 to 1858) from Escherichia coli. The purified enzyme hydrolyzed ubiquitin-7-amino-4 methylcoumarin (Ub-AMC), a general deubiquitinating enzyme substrate, with a catalytic efficiency of 13,100 M(-1)s(-1), 220-fold more efficiently than the small synthetic peptide substrate Z-LRGG-AMC, which incorporates the C-terminal four residues of ubiquitin. In addition, SARS-CoV PLpro was inhibited by the specific deubiquitinating enzyme inhibitor ubiquitin aldehyde, with an inhibition constant of 210 nM. The purified SARS-CoV PLpro disassembles branched polyubiquitin chains with lengths of two to seven (Ub2-7) or four (Ub4) units, which involves isopeptide bond cleavage. SARS-CoV PLpro processing activity was also detected against a protein fused to the C terminus of the ubiquitin-like modifier ISG15, both in vitro using the purified enzyme and in HeLa cells by coexpression with SARS-CoV PLpro (aa 1198 to 2009). These results clearly establish that SARS-CoV PLpro is a deubiquitinating enzyme, thereby confirming our earlier prediction. This unexpected activity for a coronavirus papain-like protease suggests a novel viral strategy to modulate the host cell ubiquitination machinery to its advantage. PMID- 16306592 TI - RDR6 has a broad-spectrum but temperature-dependent antiviral defense role in Nicotiana benthamiana. AB - SDE1/SGS2/RDR6, a putative RNA-dependent RNA polymerase (RdRP) from Arabidopsis thaliana, has previously been found to be indispensable for maintaining the posttranscriptional silencing of transgenes, but it is seemingly redundant for antiviral defense. To elucidate the antiviral role of this RdRP in a different host plant and to evaluate whether plant growth conditions affect its role, we down-regulated expression of the Nicotiana benthamiana homolog, NbRDR6, and examined the plants for altered susceptibility to various viruses at different growth temperatures. The results we describe here clearly show that plants with reduced expression of NbRDR6 were more susceptible to all viruses tested and that this effect was more pronounced at higher growth temperatures. Diminished expression of NbRDR6 also permitted efficient multiplication of tobacco mosaic virus in the shoot apices, leading to serious disruption with microRNA-mediated developmental regulation. Based on these results, we propose that NbRDR6 participates in the antiviral RNA silencing pathway that is stimulated by rising temperatures but suppressed by virus-encoded silencing suppressors. The relative strengths of these two factors, along with other plant defense components, critically influence the outcome of virus infections. PMID- 16306593 TI - Contributions of matrix and large protein genes of the measles virus edmonston strain to growth in cultured cells as revealed by recombinant viruses. AB - The Edmonston strain of measles virus (MV) was obtained by sequential passages of the original isolate in various cultured cells. Although attenuated in vivo, it grows efficiently in most primate cell lines. Previous studies have revealed that MV tropism cannot be solely explained by the use of CD150 and/or CD46 as a cellular receptor. In order to evaluate the contributions of individual genes of the Edmonston strain to growth in cultured cells, we generated a series of recombinant viruses in which part of the genome of the clinical isolate IC-B (which uses CD150 as a receptor) was replaced with the corresponding sequences of the Edmonston strain. The recombinant virus possessing the Edmonston hemagglutinin (H) gene (encoding the receptor-binding protein) grew as efficiently in Vero cells as the Edmonston strain. Those viruses having either the matrix (M) or large (L) protein gene from the Edmonston strain could also replicate well in Vero cells, although they entered them at low efficiencies. P64S and E89K substitutions were responsible for the ability of the M protein to make virus grow efficiently in Vero cells, while the first half of the Edmonston L gene was important for better replication. Despite efficient growth in Vero cells, the recombinant viruses with these mutations had growth disadvantage in CD150-positive lymphoid B95a cells. Thus, not only the H gene but also the M and L genes contribute to efficient replication of the Edmonston strain in some cultured cells. PMID- 16306594 TI - Modulation of HLA-G expression in human neural cells after neurotropic viral infections. AB - HLA-G is a nonclassical human major histocompatibility complex class I molecule. It may promote tolerance, leading to acceptance of the semiallogeneic fetus and tumor immune escape. We show here that two viruses-herpes simplex virus type 1 (HSV-1), a neuronotropic virus inducing acute infection and neuron latency; and rabies virus (RABV), a neuronotropic virus triggering acute neuron infection upregulate the neuronal expression of several HLA-G isoforms, including HLA-G1 and HLA-G5, the two main biologically active isoforms. RABV induces mostly HLA G1, and HSV-1 induces mostly HLA-G3 and HLA-G5. HLA-G expression is upregulated in infected cells and neighboring uninfected cells. Soluble mediators, such as beta interferon (IFN-beta) and IFN-gamma, upregulate HLA-G expression in uninfected cells. The membrane-bound HLA-G1 isoform was detected on the surface of cultured RABV-infected neurons but not on the surface of HSV-1-infected cells. Thus, neuronotropic viruses that escape the host immune response totally (RABV) or partially (HSV-1) regulate HLA-G expression on human neuronal cells differentially. HLA-G may therefore be involved in the escape of certain viruses from the immune response in the nervous system. PMID- 16306595 TI - Novel caprine adeno-associated virus (AAV) capsid (AAV-Go.1) is closely related to the primate AAV-5 and has unique tropism and neutralization properties. AB - Preexisting humoral immunity to adeno-associated virus (AAV) vectors may limit their clinical utility in gene delivery. We describe a novel caprine AAV (AAV Go.1) capsid with unique biological properties. AAV-Go.1 capsid was cloned from goat-derived adenovirus preparations. Surprisingly, AAV-Go.1 capsid was 94% identical to the human AAV-5, with differences predicted to be largely on the surface and on or under the spike-like protrusions. In an in vitro neutralization assay using human immunoglobulin G (IgG) (intravenous immune globulin [IVIG]), AAV-Go.1 had higher resistance than AAV-5 (100-fold) and resistance similar to that of AAV-4 or AAV-8. In an in vivo model, SCID mice were pretreated with IVIG to generate normal human IgG plasma levels prior to the administration of AAV human factor IX vectors. Protein expression after intramuscular administration of AAV-Go.1 was unaffected in IVIG-pretreated mice, while it was reduced 5- and 10 fold after administration of AAV-1 and AAV-8, respectively. In contrast, protein expression after intravenous administration of AAV-Go.1 was reduced 7.1-fold, similar to the 3.8-fold reduction observed after AAV-8 administration in IVIG pretreated mice, and protein expression was essentially extinguished after AAV-2 administration in mice pretreated with much less IVIG (15-fold). AAV-Go.1 vectors also demonstrated a marked tropism for lung when administered intravenously in SCID mice. The pulmonary tropism and high neutralization resistance to human preexisting antibodies suggest novel therapeutic uses for AAV-Go.1 vectors, including targeting diseases such as cystic fibrosis. Nonprimate sources of AAVs may be useful to identify additional capsids with distinct tropisms and high resistance to neutralization by human preexisting antibodies. PMID- 16306596 TI - Genetic analysis of influenza virus NS1 gene: a temperature-sensitive mutant shows defective formation of virus particles. AB - To perform a genetic analysis of the influenza A virus NS1 gene, a library of NS1 mutants was generated by PCR-mediated mutagenesis. A collection of mutant ribonucleic proteins containing the nonstructural genes was generated from the library that were rescued for an infectious virus mutant library by a novel RNP competition virus rescue procedure. Several temperature-sensitive (ts) mutant viruses were obtained by screening of the mutant library, and the sequences of their NS1 genes were determined. Most of the mutations identified led to amino acid exchanges and concentrated in the N-terminal region of the protein, but some of them occurred in the C-terminal region. Mutant 11C contained three mutations that led to amino acid exchanges, V18A, R44K, and S195P, all of which were required for the ts phenotype, and was characterized further. Several steps in the infection were slightly altered: (i) M1, M2, NS1, and neuraminidase (NA) accumulations were reduced and (ii) NS1 protein was retained in the nucleus in a temperature-independent manner, but these modifications could not justify the strong virus titer reduction at restrictive temperature. The most dramatic phenotype was the almost complete absence of virus particles in the culture medium, in spite of normal accumulation and nucleocytoplasmic export of virus RNPs. The function affected in the 11C mutant was required late in the infection, as documented by shift-up and shift-down experiments. The defect in virion production was not due to reduced NA expression, as virus yield could not be rescued by exogenous neuraminidase treatment. All together, the analysis of 11C mutant phenotype may indicate a role for NS1 protein in a late event in virus morphogenesis. PMID- 16306597 TI - Specific binding of Autographa californica M nucleopolyhedrovirus occlusion derived virus to midgut cells of Heliothis virescens larvae is mediated by products of pif genes Ac119 and Ac022 but not by Ac115. AB - Per os infectivity factors PIF1 (Ac119) and PIF2 (Ac022), like P74, are essential for oral infection of lepidopteran larval hosts of Autographa californica M nucleopolyhedrovirus (AcMNPV). Here we show that Ac115 also is a PIF (PIF3) and that, unlike PIF1 and PIF2, it does not mediate specific binding of AcMNPV occlusion-derived virus (ODV) to midgut target cells. We used an improved in vivo fluorescence dequenching assay to compare binding, fusion, and competition among control AcMNPV ODV and the ODVs of AcMNPV PIF1, PIF2, and PIF3 deletion mutants. Our results showed that binding and fusion of PIF1 and PIF2 mutants, but not the PIF3 mutant, were both qualitatively and quantitatively different from those of control ODV. Unlike control and PIF3-deficient ODV, an excess of PIF1- or PIF2 deficient ODV failed to compete effectively with control ODV's binding to specific receptors on midgut epithelial cells. Moreover, the levels of PIF1- and PIF2-deficient ODV binding were depressed threefold compared to control levels. Binding, fusion, and competition by PIF3-deficient ODV, however, were all indistinguishable from those of control ODV. These results implicated PIF1 and PIF2 as ODV envelope attachment proteins that mediate specific binding to primary target cells within the midgut. In contrast, PIF3 mediates another unidentified, but critical, early event during primary infection. PMID- 16306598 TI - Phylogenetic analysis of the main neutralization and hemagglutination determinants of all human adenovirus prototypes as a basis for molecular classification and taxonomy. AB - Human adenoviruses (HAdV) are responsible for a wide spectrum of diseases. The neutralization epsilon determinant (loops 1 and 2) and the hemagglutination gamma determinant are relevant for the taxonomy of HAdV. Precise type identification of HAdV prototypes is crucial for detection of infection chains and epidemiology. epsilon and gamma determinant sequences of all 51 HAdV were generated to propose molecular classification criteria. Phylogenetic analysis of epsilon determinant sequences demonstrated sufficient genetic divergence for molecular classification, with the exception of HAdV-15 and HAdV-29, which also cannot be differentiated by classical cross-neutralization. Precise sequence divergence criteria for typing (<2.5% from loop 2 prototype sequence and <2.4% from loop 1 sequence) were deduced from phylogenetic analysis. These criteria may also facilitate identification of new HAdV prototypes. Fiber knob (gamma determinant) phylogeny indicated a two-step model of species evolution and multiple intraspecies recombination events in the origin of HAdV prototypes. HAdV-29 was identified as a recombination variant of HAdV-15 (epsilon determinant) and a speculative, not-yet-isolated HAdV prototype (gamma determinant). Subanalysis of molecular evolution in hypervariable regions 1 to 6 of the epsilon determinant indicated different selective pressures in subclusters of species HAdV-D. Additionally, gamma determinant phylogenetic analysis demonstrated that HAdV-8 did not cluster with -19 and -37 in spite of their having the same tissue tropism. The phylogeny of HAdV-E4 suggested origination by interspecies recombination between HAdV-B (hexon) and HAdV-C (fiber), as in simian adenovirus 25, indicating additional zoonotic transfer. In conclusion, molecular classification by systematic sequence analysis of immunogenic determinants yields new insights into HAdV phylogeny and evolution. PMID- 16306599 TI - Episomal maintenance of plasmids with hybrid origins in mouse cells. AB - Bovine papillomavirus type 1 (BPV1), Epstein-Barr virus (EBV), and human herpesvirus 8 genomes are stably maintained as episomes in dividing host cells during latent infection. The mitotic segregation/partitioning function of these episomes is dependent on single viral protein with specific DNA-binding activity and its multimeric binding sites in the viral genome. In this study we show that, in the presence of all essential viral trans factors, the segregation/partitioning elements from both BPV1 and EBV can provide the stable maintenance function to the mouse polyomavirus (PyV) core origin plasmids but fail to do so in the case of complete PyV origin. Our study is the first which follows BPV1 E2- and minichromosome maintenance element (MME)-dependent stable maintenance function with heterologous replication origins. In mouse fibroblast cell lines expressing PyV large T antigen (LT) and either BPV1 E2 or EBV EBNA1, the long-term episomal replication of plasmids carrying the PyV minimal origin together with the MME or family of repeats (FR) element can be monitored easily for 1 month under nonselective conditions. Our data demonstrate clearly that the PyV LT-dependent replication function and the segregation/partitioning function of the BPV1 or EBV are compatible in certain, but not all, configurations. The quantitative analysis indicates a loss rate of 6% per cell, doubling in the case of MME-dependent plasmids, and 13% in the case of FR-dependent plasmids in nonselective conditions. Our data clearly indicate that maintenance functions from different viruses are principally interexchangeable and can provide a segregation/partitioning function to different heterologous origins in a variety of cells. PMID- 16306600 TI - Th-cytotoxic T-lymphocyte chimeric epitopes extended by Nepsilon-palmitoyl lysines induce herpes simplex virus type 1-specific effector CD8+ Tc1 responses and protect against ocular infection. AB - Molecularly defined vaccine formulations capable of inducing antiviral CD8+ T cell-specific immunity in a manner compatible with human delivery are limited. Few molecules achieve this target without the support of an appropriate immunological adjuvant. In this study, we investigate the potential of totally synthetic palmitoyl-tailed helper-cytotoxic-T-lymphocyte chimeric epitopes (Th CTL chimeric lipopeptides) to induce herpes simplex virus type 1 (HSV-1)-specific CD8+ T-cell responses. As a model antigen, the HSV-1 glycoprotein B498-505 (gB498 505) CD8+ CTL epitope was synthesized in line with the Pan DR peptide (PADRE), a universal CD4+ Th epitope. The peptide backbone, composed solely of both epitopes, was extended by N-terminal attachment of one (PAM-Th-CTL), two [(PAM)2 Th-CTL], or three [(PAM)3-Th-CTL] palmitoyl lysines and delivered to H2b mice in adjuvant-free saline. Potent HSV-1 gB498-505-specific antiviral CD8+ T-cell effector type 1 responses were induced by each of the palmitoyl-tailed Th-CTL chimeric epitopes, irrespective of the number of lipid moieties. The palmitoyl tailed Th-CTL chimeric epitopes provoked cell surface expression of major histocompatibility complex and costimulatory molecules and production of interleukin-12 and tumor necrosis factor alpha proinflammatory cytokines by immature dendritic cells. Following ocular HSV-1 challenge, palmitoyl-tailed Th CTL-immunized mice exhibited a decrease of virus replication in the eye and in the local trigeminal ganglion and reduced herpetic blepharitis and corneal scarring. The rational of the molecularly defined vaccine approach presented in this study may be applied to ocular herpes and other viral infections in humans, providing steps are taken to include appropriate Th and CTL epitopes and lipid groups. PMID- 16306601 TI - Respiratory syncytial virus-inducible BCL-3 expression antagonizes the STAT/IRF and NF-kappaB signaling pathways by inducing histone deacetylase 1 recruitment to the interleukin-8 promoter. AB - Respiratory syncytial virus (RSV) is a paramyxovirus that produces airway inflammation, in part by inducing interleukin-8 (IL-8) expression, a CXC-type chemokine, via the NF-kappaB/RelA and STAT/IRF signaling pathways. In RSV infected A549 cells, IL-8 transcription attenuates after 24 h in spite of ongoing viral replication and persistence of nuclear RelA, suggesting a mechanism for transcriptional attenuation. RSV infection induces B-cell lymphoma protein -3 (Bcl-3) expression 6 to 12 h after viral infection, at times when IL-8 transcription is inhibited. By contrast, 293 cells, deficient in inducible Bcl-3 expression, show no attenuation of IL-8 transcription. We therefore examined Bcl 3's role in terminating virus-inducible IL-8 transcription. Transient expression of Bcl-3 potently inhibited virus-inducible IL-8 transcription by disrupting both the NF-kappaB and STAT/IRF pathways. Although previously Bcl-3 was thought to capture 50-kDa NF-kappaB1 isoforms in the cytoplasm, immunoprecipitation (IP) and electrophoretic mobility shift assays indicate that nuclear Bcl-3 associates with NF-kappaB1 without affecting DNA binding. Additionally, Bcl-3 potently inhibited the STAT/IRF pathway. Nondenaturing co-IP assays indicate that nuclear Bcl-3 associates with STAT-1 and histone deacetylase 1 (HDAC-1), increasing HDAC-1 recruitment to the IL-8 promoter. Treatment with the HDAC inhibitor trichostatin A blocks attenuation of IL-8 transcription. A nuclear targeting-deficient Bcl-3 is unable to enhance HDAC-1-mediated chemokine repression. Finally, small inhibitory RNA-mediated Bcl-3 "knockdown" resulted in enhanced RSV-induced chemokine expression in A549 cells. These data indicate that Bcl-3 is a virus inducible inhibitor of chemokine transcription by interfering with the NF-kappaB and STAT/IRF signaling pathways by complexing with them and recruiting HDAC-1 to attenuate target promoter activity. PMID- 16306602 TI - Soluble receptor-mediated targeting of mouse hepatitis coronavirus to the human epidermal growth factor receptor. AB - The mouse hepatitis coronavirus (MHV) infects murine cells by binding of its spike (S) protein to murine CEACAM1a. The N-terminal part of this cellular receptor (soR) is sufficient for S binding and for subsequent induction of the conformational changes required for virus-cell membrane fusion. Here we analyzed whether these characteristics can be used to redirect MHV to human cancer cells. To this end, the soR domain was coupled to single-chain monoclonal antibody 425, which is directed against the human epidermal growth factor receptor (EGFR), resulting in a bispecific adapter protein (soR-425). The soR and soR-425 proteins, both produced with the vaccinia virus system, were able to neutralize MHV infection of murine LR7 cells. However, only soR-425 was able to target MHV to human EGFR-expressing cancer cells. Interestingly, the targeted infections induced syncytium formation. Furthermore, the soR-425-mediated infections were blocked by heptad repeat-mimicking peptides, indicating that virus entry requires the regular S protein fusion process. We conclude that the specific spike-binding property of the CEACAM1a N-terminal fragment can be exploited to direct the virus to selected cells by linking it to a moiety able to bind a receptor on those cells. This approach might be useful in the development of tumor-targeted coronaviruses. PMID- 16306603 TI - Genomic sequence analysis of Epstein-Barr virus strain GD1 from a nasopharyngeal carcinoma patient. AB - To date, the only entire Epstein-Barr virus (EBV) genomic sequence available in the database is the prototype B95.8, which was derived from an individual with infectious mononucleosis. A causative link between EBV and nasopharyngeal carcinoma (NPC), a disease with a distinctly high incidence in southern China, has been widely investigated. However, no full-length analysis of any substrain of EBV from this area has been reported. In this study, we analyzed the entire genomic sequence of an EBV strain from a patient with NPC in Guangdong, China. This EBV strain was termed GD1 (Guangdong strain 1), and the full-length sequence of GD1 was submitted to the GenBank database. The assigned accession number is AY961628. The entire GD1 sequence is 171,656 bp in length, with 59.5% G+C content and 40.5% A+T content. We detected many sequence variations in GD1 compared to prototypical strain B95.8, including 43 deletion sites, 44 insertion sites, and 1,413 point mutations. Furthermore, we evaluated the frequency of some of these GD1 mutations in Cantonese NPC patients and found them to be highly prevalent. These findings suggest that GD1 is highly representative of the EBV strains isolated from NPC patients in Guangdong, China, an area with the highest incidence of NPC in the world. Furthermore, these findings provide the second full-length sequence analysis of any EBV strain as well as the first full-length sequence analysis of an NPC-derived EBV strain. PMID- 16306604 TI - Basic residues in hypervariable region 1 of hepatitis C virus envelope glycoprotein e2 contribute to virus entry. AB - The N terminus of hepatitis C virus (HCV) envelope glycoprotein E2 contains a hypervariable region (HVR1) which has been proposed to play a role in viral entry. Despite strong amino acid variability, HVR1 is globally basic, with basic residues located at specific sequence positions. Here we show by analyzing a large number of HVR1 sequences that the frequency of basic residues at each position is genotype dependent. We also used retroviral pseudotyped particles (HCVpp) harboring genotype 1a envelope glycoproteins to study the role of HVR1 basic residues in entry. Interestingly, HCVpp infectivity globally increased with the number of basic residues in HVR1. However, a shift in position of some charged residues also modulated HCVpp infectivity. In the absence of basic residues, infectivity was reduced to the same level as that of a mutant deleted of HVR1. We also analyzed the effect of these mutations on interactions with some potential HCV receptors. Recognition of CD81 was not affected by changes in the number of charged residues, and we did not find a role for heparan sulfates in HCVpp entry. The involvement of the scavenger receptor class B type I (SR-BI) was indirectly analyzed by measuring the enhancement of infectivity of the mutants in the presence of the natural ligand of SR-BI, high-density lipoproteins (HDL). However, no correlation between the number of basic residues within HVR1 and HDL enhancement effect was observed. Despite the lack of evidence of the involvement of known potential receptors, our results demonstrate that the presence of basic residues in HVR1 facilitates virus entry. PMID- 16306605 TI - Upregulation of protein phosphatase 2Ac by hepatitis C virus modulates NS3 helicase activity through inhibition of protein arginine methyltransferase 1. AB - Hepatitis C virus (HCV) is a major cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma worldwide. HCV has a positive-strand RNA genome of about 9.4 kb in size, which serves as a template for replication and for translation of a polyprotein of about 3,000 amino acids. The polyprotein is cleaved co- and posttranslationally by cellular and viral proteases into at least 10 different mature proteins. One of these proteins, nonstructural protein 3 (NS3), has serine protease and NTPase/RNA helicase activity. Arginine 467 in the helicase domain of NS3 (arginine 1493 in the polyprotein) can be methylated by protein arginine methyltransferase 1 (PRMT1). Here we report that the methylation of NS3 inhibits the enzymatic activity of the helicase. Furthermore, we found that PRMT1 activity itself is regulated by protein phosphatase 2A (PP2A). PP2A inhibits PRMT1 enzymatic activity and therefore increases the helicase activity of NS3. This is important, because we found an increased expression of PP2A in cell lines with inducible HCV protein expression, in transgenic mice expressing HCV proteins in hepatocytes, and in liver biopsy samples from patients with chronic hepatitis C. Interestingly, up-regulation of PP2A not only modulates the enzymatic activity of an important viral protein, NS3 helicase, but also interferes with the cellular defense against viruses by inhibiting interferon-induced signaling through signal transducer and activator of transcription 1 (STAT1). We conclude that up regulation of PP2A might be crucial for the efficient replication of HCV and propose PP2A as a potential target for anti-HCV treatment strategies. PMID- 16306606 TI - A homozygous nonsense mutation (428G-->A) in the human secretor (FUT2) gene provides resistance to symptomatic norovirus (GGII) infections. AB - Noroviruses (formerly Norwalk-like viruses) are a major cause of acute gastroenteritis worldwide and are associated with a significant number of nosocomial and food-borne outbreaks. In this study we show that the human secretor FUT2 gene, which codes for an alpha(1,2)-fucosyltransferase synthesizing the H-type 1 antigen in saliva and mucosa, is associated with susceptibility to norovirus infections. Allelic polymorphism characterization at nucleotide 428 for symptomatic (n = 53) and asymptomatic (n = 62) individuals associated with nosocomial and sporadic norovirus outbreaks revealed that homozygous nonsense mutation (428G-->A) in FUT2 segregated with complete resistance for the disease. Of all symptomatic individuals, 49% were homozygous (SeSe) and 51% heterozygous (Sese428) secretors, and none were secretor negative (se428se428), in contrast to 20% nonsecretors (se428se428) among Swedish blood donors (n = 104) (P < 0.0002) and 29% for asymptomatic individuals associated with nosocomial outbreaks (P < 0.00001). Furthermore, saliva from secretor-positive and symptomatic patients but not from secretor-negative and asymptomatic individuals bound the norovirus strain responsible for that particular outbreak. This is the first report showing that the FUT2 nonsecretor (se428se428) genotype is associated with resistance to nosocomial and sporadic outbreaks with norovirus. PMID- 16306607 TI - A combination DNA and attenuated simian immunodeficiency virus vaccine strategy provides enhanced protection from simian/human immunodeficiency virus-induced disease. AB - Among the most effective vaccine candidates tested in the simian immunodeficiency virus (SIV)/macaque system, live attenuated viruses have been shown to provide the best protection from challenge. To investigate if preimmunization would increase the level of protection afforded by live attenuated SIVmac239Deltanef (Deltanef), macaques were given two priming immunizations of DNA encoding SIV Gag and Pol proteins, with control macaques receiving vector DNA immunizations. In macaques receiving the SIV DNA inoculation, SIV-specific cellular but not humoral responses were readily detectable 2 weeks after the second DNA inoculation. Following boosting with live attenuated virus, control of Deltanef replication was superior in SIV-DNA-primed macaques versus vector-DNA-primed macaques and was correlated with higher levels of CD8+/gamma-interferon-positive and/or interleukin-2-positive cells. Challenge with an intravenous inoculation of simian/human immunodeficiency virus (SHIV) strain SHIV89.6p resulted in infection of all animals. However, macaques receiving SIV DNA as the priming immunizations had statistically lower viral loads than control animals and did not develop signs of disease, whereas three of seven macaques receiving vector DNA showed severe CD4+ T-cell decline, with development of AIDS in one of these animals. No correlation of immune responses to protection from disease could be derived from our analyses. These results demonstrate that addition of a DNA prime to a live attenuated virus provided better protection from disease following challenge than live attenuated virus alone. PMID- 16306608 TI - Genetic and stochastic influences on the interaction of human immunodeficiency virus type 1 and cytotoxic T lymphocytes in identical twins. AB - Human immunodeficiency virus type 1 (HIV-1) evolves in vivo under selective pressure from CD8+ T-lymphocyte (CTL) responses, which are in turn determined by host and viral genetic factors, such as restricting major histocompatibility complex molecules and the available viral epitope sequences. However, CTL are derived stochastically through the random gene rearrangements to produce T-cell receptors (TCR), and the relative impact of genetic versus stochastic processes on CTL targeting of HIV and immune-driven viral evolution is unclear. Here we evaluate identical twins infected with HIV-1 as neonates from a common blood transfusion, with subsequently similar environmental exposures, thereby allowing controlled comparisons of CTL targeting and viral evolution. Seventeen years after infection, their CTL targeting of HIV-1 was remarkably similar. In contrast, their overall TCR profiles were highly dissimilar, and a dominant epitope was recognized by distinctly different TCR in each twin. Furthermore, their viral epitopes had diverged, and there was ongoing viral phylogenetic divergence between the twins between 12 and 17 years after infection. These results indicate that while CTL targeting is predominately genetically determined, stochastic influences render the interaction of HIV-1 and host immunity, and therefore viral escape and CTL efficacy, unpredictable. PMID- 16306609 TI - Division of labor within human immunodeficiency virus integrase complexes: determinants of catalysis and target DNA capture. AB - Following the completion of reverse transcription, the human immunodeficiency virus integrase (IN) enzyme covalently links the viral cDNA to a host cell chromosome. An IN multimer carries out this reaction, but the roles of individual monomers within the complex are mostly unknown. Here we analyzed the distribution of functions for target DNA capture and catalysis within the IN multimer. We used forced complementation between pairs of IN deletion derivatives in vitro as a tool for probing cis-trans relationships and analyzed amino acid substitutions affecting either catalysis or target site selection within these complementing complexes. This allowed the demonstration that the IN variant contributing the active catalytic domain was also responsible for recognition of the integration target DNA. We were further able to establish that a single monomer is responsible for both functions by use of assay mixtures containing three different IN genotypes. These data specify the ligands bound at the catalytically relevant IN monomer and allow more-specific modeling of the mechanism of inhibitors that also bind this surface of IN. PMID- 16306610 TI - Production of infectious human cytomegalovirus virions is inhibited by drugs that disrupt calcium homeostasis in the endoplasmic reticulum. AB - We previously reported that human cytomegalovirus (HCMV) infection induces endoplasmic reticulum (ER) stress, resulting in activation of the unfolded protein response (UPR). Although some normal consequences of UPR activation (e.g., translation attenuation) are detrimental to viral infection, we have previously shown that HCMV infection adapts the UPR to benefit the viral infection (14). For example, UPR-induced translation attenuation is inhibited by viral infection, while potentially beneficial aspects of the UPR are maintained. In the present work, we tested the ability of HCMV to overcome a robust induction of the UPR by the drugs thapsigargin and clotrimazole (CLT), which disrupt ER calcium homeostasis. A 24-h treatment with these drugs beginning at 48, 72, or 96 h postinfection (hpi) completely inhibited further production of infectious virions. HCMV could not overcome the inhibition of global translation by CLT; however, between 48 and 72 hpi, HCMV overcame translational inhibition caused by thapsigargin. Despite the restoration of translation in thapsigargin, the accumulation of immediate-early and early gene products was modestly retarded (50% or less), whereas the accumulation of an early-late and late gene product was significantly retarded. Electron microscopic analysis shows that the drugs severely disrupt the maturation of HCMV virions. This can be accounted for by both the retarded accumulation of late gene products and the drug-induced depletion of ER calcium, which disrupts critical cellular functions needed for maturation. PMID- 16306611 TI - Unique ligand binding sites on CXCR4 probed by a chemical biology approach: implications for the design of selective human immunodeficiency virus type 1 inhibitors. AB - The chemokine receptor CXCR4 plays an important role as the receptor for the normal physiological function of stromal cell-derived factor 1alpha (SDF-1alpha) and the coreceptor for the entry of human immunodeficiency virus type 1 (HIV-1) into the cell. In a recent work (S. Tian et al., J. Virol. 79:12667-12673, 2005), we found that many residues throughout CXCR4 transmembrane (TM) and extracellular loop 2 domains are specifically involved in interaction with HIV-1 gp120, as most of these sites did not play a role in either SDF-1alpha binding or signaling. These results provided direct experimental evidence for the distinct functional sites on CXCR4 for HIV-1 and the normal ligand SDF-1alpha. To further understand the CXCR4-ligand interaction and to develop new CXCR4 inhibitors to block HIV-1 entry, we have recently generated a new family of unnatural chemokines, termed synthetically and modularly modified (SMM) chemokines, derived from the native sequence of SDF-1alpha or viral macrophage inflammatory protein II (vMIP-II). These SMM chemokines contain various de novo-designed sequence replacements and substitutions by d-amino acids and display more enhanced CXCR4 selectivity, binding affinities, and/or anti-HIV activities than natural chemokines. Using these novel CXCR4-targeting SMM chemokines as receptor probes, we conducted ligand binding site mapping experiments on a panel of site-directed mutants of CXCR4. Here, we provide the first experimental evidence demonstrating that SMM chemokines interact with many residues on CXCR4 TM and extracellular domains that are important for HIV-1 entry, but not SDF-1alpha binding or signaling. The preferential overlapping in the CXCR4 binding residues of SMM chemokines with HIV 1 over SDF-1alpha illustrates a mechanism for the potent HIV-1 inhibition by these SMM chemokines. The discovery of distinct functional sites or conformational states influenced by these receptor sites mediating different functions of the natural ligand versus the viral or synthetic ligands has important implications for drug discovery, since the sites shared by SMM chemokines and HIV-1 but not by SDF-1alpha can be targeted for the development of selective HIV-1 inhibitors devoid of interference with normal SDF-1alpha function. PMID- 16306612 TI - Overexpression of tumor necrosis factor alpha by a recombinant rabies virus attenuates replication in neurons and prevents lethal infection in mice. AB - The effect of tumor necrosis factor alpha (TNF-alpha) on rabies virus (RV) infection of the mouse central nervous system (CNS) was studied, using recombinant RV engineered to express either soluble TNF-alpha [SPBN-TNF-alpha+] or insoluble membrane-bound TNF-alpha [SPBN-TNF-alpha(MEM)]. Growth curves derived from infections of mouse neuroblastoma NA cells revealed significantly less spread and production of SPBN-TNF-alpha+ than of SPBN-TNF-alpha(MEM) or SPBN TNF-alpha-, which carries an inactivated TNF-alpha gene. The expression of soluble or membrane-bound TNF-alpha was not associated with increased cell death or induction of alpha/beta interferons. Brains of mice infected intranasally with SPBN-TNF-alpha+ showed significantly less virus spread than did mouse brains after SPBN-TNF-alpha- infection, and none of the SPBN-TNF-alpha+-infected mice succumbed to RV infection, whereas 80% of SPBN-TNF-alpha- -infected mice died. Reduced virus spread in SPBN-TNF-alpha+-infected mouse brains was paralleled by enhanced CNS inflammation, including T-cell infiltration and microglial activation. These data suggest that TNF-alpha exerts its protective activity in the brain directly through an as yet unknown antiviral mechanism and indirectly through the induction of inflammatory processes in the CNS. PMID- 16306613 TI - SOCS1 and SOCS3 are targeted by hepatitis C virus core/gC1qR ligation to inhibit T-cell function. AB - T cells play an important role in the control of hepatitis C virus (HCV) infection. We have previously demonstrated that the HCV core inhibits T-cell responses through interaction with gC1qR. We show here that core proteins from chronic and resolved HCV patients differ in sequence, gC1qR-binding ability, and T-cell inhibition. Specifically, chronic core isolates bind to gC1qR more efficiently and inhibit T-cell proliferation as well as gamma interferon (IFN gamma) production more profoundly than resolved core isolates. This inhibition is mediated by the disruption of STAT phosphorylation through the induction of SOCS molecules. Silencing either SOCS1 or SOCS3 by small interfering RNA dramatically augments the production of IFN-gamma in T cells, thereby abrogating the inhibitory effect of core. Additionally, the ability of core proteins from patients with chronic infections to induce SOCS proteins and suppress STAT activation greatly exceeds that of core proteins from patients with resolved infections. These results suggest that the HCV core/gC1qR-induced T-cell dysfunction involves the induction of SOCS, a powerful inhibitor of cytokine signaling, which represents a novel mechanism by which a virus usurps the host machinery for persistence. PMID- 16306614 TI - Epstein-Barr-virus-encoded LMP2A induces primary epithelial cell migration and invasion: possible role in nasopharyngeal carcinoma metastasis. AB - Nonkeratinizing nasopharyngeal carcinomas (NPC) are >95% associated with the expression of the Epstein-Barr virus (EBV) LMP2A latent protein. However, the role of EBV, in particular, LMP2A, in tumor progression is not well understood. Using Affymetrix chips and a pattern-matching computational technique (neighborhood analysis), we show that the level of LMP2A expression in NPC biopsy samples correlates with that of a cellular protein, integrin-alpha-6 (ITGalpha6), that is associated with cellular migration in vitro and metastasis in vivo. We have recently developed a primary epithelial model from tonsil tissue to study EBV infection in epithelial cells. Here we report that LMP2A expression in primary tonsil epithelial cells causes them to become migratory and invasive, that ITGalpha6 RNA levels are up-regulated in epithelial cells expressing LMP2, and that ITGalpha6 protein levels are increased in the migrating cells. Blocking antibodies against ITGalpha6 abrogated LMP2-induced invasion through Matrigel by primary epithelial cells. Our results provide a link between LMP2A expression, ITGalpha6 expression, epithelial cell migration, and NPC metastasis and suggest that EBV infection may contribute to the high incidence of metastasis in NPC progression. PMID- 16306615 TI - Activation of the ATR pathway by human immunodeficiency virus type 1 Vpr involves its direct binding to chromatin in vivo. AB - The human immunodeficiency virus type 1 (HIV-1) protein Vpr (viral protein R) arrests cells in the G2 phase of the cell cycle, a process that requires activation of the ATR (ataxia-telangiectasia and Rad3-related) pathway. In this study we demonstrate that the expression of Vpr does not cause DNA double-strand breaks but rather induces ATR activation, as indicated by induction of Chk1 phosphorylation and the formation of gamma-H2AX and 53BP1 nuclear foci. We define a C-terminal domain containing repeated H(F/S)RIG sequences required for Vpr induced activation of ATR. Further investigation of the mechanism by which Vpr activates the ATR pathway reveals an increase in chromatin binding of replication protein A (RPA) upon Vpr expression. Immunostaining shows that RPA localizes to nuclear foci in Vpr-expressing cells. Furthermore, we demonstrate direct binding of Vpr to chromatin in vivo, whereas Vpr C-terminal domain mutants lose this chromatin-binding activity. These data support a mechanism whereby HIV-1 Vpr induces ATR activation by targeting the host cell DNA and probably interfering with normal DNA replication. PMID- 16306616 TI - Baculovirus entry into human hepatoma cells. AB - Autographa californica multiple nucleopolyhedrovirus (AcMNPV), a prototype member of the Baculoviridae family, has gained increasing interest as a potential vector candidate for mammalian gene delivery applications. AcMNPV is known to enter both dividing and nondividing mammalian cell lines in vitro, but the mode and kinetics of entry as well as the intracellular transport of the virus in mammalian cells is poorly understood. The general objective of this study was to characterize the entry steps of AcMNPV- and green fluorescent protein-displaying recombinant baculoviruses in human hepatoma cells. The viruses were found to bind and transduce the cell line efficiently, and electron microscopy studies revealed that virions were located on the cell surface in pits with an electron-dense coating resembling clathrin. In addition, virus particles were found in larger noncoated plasma membrane invaginations and in intracellular vesicles resembling macropinosomes. In double-labeling experiments, virus particles were detected by confocal microscopy in early endosomes at 30 min and in late endosomes starting at 45 min posttransduction. Viruses were also seen in structures specific for early endosomal as well as late endosomal/lysosomal markers by nanogold preembedding immunoelectron microscopy. No indication of viral entry into recycling endosomes or the Golgi complex was observed by confocal microscopy. In conclusion, these results suggest that AcMNPV enters mammalian cells via clathrin mediated endocytosis and possibly via macropinocytosis. Thus, the data presented here should enable future design of baculovirus vectors suitable for more specific and enhanced delivery of genetic material into mammalian cells. PMID- 16306617 TI - New genotype of avian influenza H5N1 viruses isolated from tree sparrows in China. AB - The 2004 outbreaks of highly pathogenic avian influenza H5N1 disease in China led to a great poultry loss and society attention. A survey of avian influenza viruses was conducted on tree sparrows (Passer montanus) collected in China in 2004. Four viruses were isolated from free-living tree sparrows. The results of the whole-genome analysis indicated that an H5N1 virus with a new genotype is circulating among tree sparrows. The hemagglutinin and neuraminidase genes of the new genotype were derived from Gs/Gd/96-like viruses and the nuclear protein gene descended from the 2001 genotype A H5N1 viruses, while the other inner genes originated from an unknown influenza virus. In experimental infection, all four viruses were highly pathogenic to chickens but not pathogenic to ducks or mice. The four tree sparrow viruses were different from the 2003 tree sparrow strain (genotype Z) in Hong Kong. The results suggested that H5N1 viruses might be distributed widely in tree sparrows. PMID- 16306618 TI - Recombination in the genesis and evolution of hepatitis B virus genotypes. AB - Hepatitis B virus (HBV) infection is widely distributed in both human and ape populations throughout the world and is a major cause of human morbidity and mortality. HBV variants are currently classified into the human genotypes A to H and species-associated chimpanzee and gibbon/orangutan groups. To examine the role of recombination in the evolution of HBV, large-scale data retrieval and automated phylogenetic analysis (TreeOrder scanning) were carried out on all available published complete genome sequences of HBV. We detected a total of 24 phylogenetically independent potential recombinants (different genotype combinations or distinct breakpoints), eight of which were previously undescribed. Instances of intergenotype recombination were observed in all human and ape HBV variants, including evidence for a novel gibbon/genotype C recombinant among HBV variants from Vietnam. By recording sequence positions in trees generated from sequential fragments across the genome, violations of phylogeny between trees also provided evidence for frequent intragenotype recombination between members of genotypes A, D, F/H, and gibbon variants but not in B, C, or the Asian B/C recombinant group. In many cases, favored positions for both inter- and intragenotype recombination matched positions of phylogenetic reorganization between the human and ape genotypes, such as the end of the surface gene and the core gene, where sequence relationships between genotypes changed in the TreeOrder scan. These findings provide evidence for the occurrence of past, extensive recombination events in the evolutionary history of the currently classified genotypes of HBV and potentially in changes in its global epidemiology and associations with human disease. PMID- 16306619 TI - Human cytomegalovirus infection induces specific hyperphosphorylation of the carboxyl-terminal domain of the large subunit of RNA polymerase II that is associated with changes in the abundance, activity, and localization of cdk9 and cdk7. AB - Human cytomegalovirus infection in the presence of the cyclin-dependent kinase (cdk) inhibitor roscovitine leads to changes in differential splicing and the polyadenylation of immediate early IE1/IE2 and UL37 transcripts (V. Sanchez, A. K. McElroy, J. Yen, S. Tamrakar, C. L. Clark, R. A. Schwartz, and D. H. Spector, J. Virol. 78:11219-11232, 2004). To determine if this was associated with specific phosphorylation of the C-terminal domain (CTD) of the RNA polymerase II (RNAP II) large subunit by cdk7/cyclin H and cdk9/cyclin T1, we examined the expression and localization of these kinases and the various phosphorylated forms of RNAP II. Infection resulted in increased RNAP II CTD phosphorylated on serines 2 and 5 and increased levels of activity of cdk7 and cdk9. At early times, cdk9 localizes with input viral DNA, and aggregates of cdk9 and cdk7 and a subset of Ser2-phosphorylated RNAP II colocalize with IE1/IE2 proteins adjacent to promyelocytic leukemia protein oncogenic domains. Later, cdk9 and Ser2 phosphorylated RNAP II form a nuclear punctate pattern; cdk7 resides in replication centers, and Ser5-phosphorylated RNAP II clusters at the peripheries of replication centers. Roscovitine treatment leads to decreased levels of hyperphosphorylated RNAP II (RNAP IIo) in infected cells and of hypophosphorylated RNAP II in mock-infected and infected cells. The RNAP IIo decrease does not occur if roscovitine is added 8 h postinfection, as was previously observed for processing of IE transcripts. These results suggest that accurate IE gene expression requires specific phosphorylation of the RNAP II CTD early in infection. PMID- 16306620 TI - Human cytomegalovirus UL97 Kinase is required for the normal intranuclear distribution of pp65 and virion morphogenesis. AB - Recombinant human cytomegaloviruses that do not express UL97 kinase activity exhibit a distinctive plaque morphology characterized by the formation of highly refractile bodies late in infection. These structures were also observed in infected cells treated with the UL97 kinase inhibitor maribavir. Nuclear inclusions were purified to near homogeneity, and the constituent proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry. This analysis demonstrated that the aggregates were formed principally of the tegument proteins pp65 and ppUL25 but also contained additional virion structural proteins including the major capsid protein. Immunoblotting experiments confirmed these results and identified a number of additional viral proteins present in the purified tegument aggregates. Interestingly, the formation of these structures appeared to be dependent on pp65, since it was not induced in cells infected with a recombinant virus with this open reading frame deleted. Morphologically similar aggregates could be reproduced in nuclei of uninfected cells by overexpressing pp65, and their formation was prevented by coexpressing the UL97 kinase. Inhibition of UL97 kinase activity with maribavir or mutation of an essential amino acid in the kinase abolished its ability to prevent aggregate formation. These data taken together suggest that the UL97 kinase impacts the aggregation of pp65 in the nuclei of infected cells. We propose that the kinase plays an important role in the acquisition of tegument during virion morphogenesis in the nucleus and that this activity represents an important step in the production of mature virus particles. PMID- 16306621 TI - Phylogenetic incongruence among oncogenic genital alpha human papillomaviruses. AB - The human papillomaviruses (HPVs) have long been thought to follow a monophyletic pattern of evolution with little if any evidence for recombination between genomes. On the basis of this model, both oncogenicity and tissue tropism appear to have evolved once. Still, no systematic statistical analyses have shown whether monophyly is the rule across all HPV open reading frames (ORFs). We conducted a taxonomic analysis of 59 mucosal/genital HPVs using whole-genome and sliding-window similarity measures; maximum-parsimony, neighbor-joining, and Bayesian phylogenetic analyses; and localized incongruence length difference (LILD) analyses. The algorithm for the LILD analyses localized incongruence by calculating the tree length differences between constrained and unconstrained nodes in a total-evidence tree across all HPV ORFs. The process allows statistical evaluation of every ORF/node pair in the total-evidence tree. The most significant incongruence was observed at the putative high-risk (i.e., cancer-associated) node, the common oncogenic ancestor for alpha HPV species 9 (e.g., HPV type 16 [HPV16]), 11, 7 (e.g., HPV18), 5, and 6. Although these groups share early-gene homology, including high degrees of similarity among E6 and E7, groups 9 and 11 diverge from groups 7, 5, and 6 with respect to L2 and L1. The HPV species groups primarily associated with cervical and anogenital cancers appear to follow two distinct evolutionary paths, one conferred by the early genes and another by the late genes. The incongruence in the genital HPV phylogeny could have occurred from an early recombination event, an ecological niche change, and/or asymmetric genome convergence driven by intense selection. These data indicate that the phylogeny of the oncogenic HPVs is complex and that their evolution may not be monophyletic across all genes. PMID- 16306622 TI - Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: role of ciliated cells in viral spread in the conducting airways of the lungs. AB - Severe acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and in vitro models of the lung are needed to elucidate cellular targets and the consequences of viral infection. The SARS-CoV receptor, human angiotensin 1 converting enzyme 2 (hACE2), was detected in ciliated airway epithelial cells of human airway tissues derived from nasal or tracheobronchial regions, suggesting that SARS-CoV may infect the proximal airways. To assess infectivity in an in vitro model of human ciliated airway epithelia (HAE) derived from nasal and tracheobronchial airway regions, we generated recombinant SARS-CoV by deletion of open reading frame 7a/7b (ORF7a/7b) and insertion of the green fluorescent protein (GFP), resulting in SARS-CoV GFP. SARS-CoV GFP replicated to titers similar to those of wild-type viruses in cell lines. SARS-CoV specifically infected HAE via the apical surface and replicated to titers of 10(7) PFU/ml by 48 h postinfection. Polyclonal antisera directed against hACE2 blocked virus infection and replication, suggesting that hACE2 is the primary receptor for SARS CoV infection of HAE. SARS-CoV structural proteins and virions localized to ciliated epithelial cells. Infection was highly cytolytic, as infected ciliated cells were necrotic and shed over time onto the luminal surface of the epithelium. SARS-CoV GFP also replicated to a lesser extent in ciliated cell cultures derived from hamster or rhesus monkey airways. Efficient SARS-CoV infection of ciliated cells in HAE provides a useful in vitro model of human lung origin to study characteristics of SARS-CoV replication and pathogenesis. PMID- 16306623 TI - Signal peptide cleavage and internal targeting signals direct the hepatitis C virus p7 protein to distinct intracellular membranes. AB - The hepatitis C virus (HCV) p7 protein forms an amantadine-sensitive ion channel required for viral replication in chimpanzees, though its precise role in the life cycle of HCV is unknown. In an attempt to gain some insights into p7 function, we examined the intracellular localization of p7 using epitope tags and an anti-p7 peptide antibody, antibody 1055. Immunofluorescence labeling of p7 at its C terminus revealed an endoplasmic reticulum (ER) localization independent of the presence of its signal peptide, whereas labeling the N terminus gave a mitochondrial-type distribution in brightly labeled cells. Both of these patterns could be visualized within individual cells, suggestive of separate pools of p7 where the N and C termini differed in accessibility to antibody. These patterns were disrupted by preventing signal peptide cleavage. Subcellular fractionation revealed that p7 was enriched in a heavy membrane fraction associated with mitochondria as well as normal ER-derived microsomes. The complex regulation of the intracellular distribution of p7 suggests that p7 plays multiple roles in the HCV life cycle either intracellularly or as a virion component. PMID- 16306624 TI - The C terminus of the nucleoprotein of influenza A virus delivers antigens transduced by Tat to the trans-golgi network and promotes an efficient presentation through HLA class I. AB - Cytotoxic T lymphocytes (CTLs) are the most powerful weapon of the immune system to eliminate cells infected by intracellular parasites or tumors. However, very often, escape mechanisms overcome CTL immune surveillance by impairing the classical HLA class I antigen-processing pathway. Here, we describe a strategy for CTL activation based on the ability of Tat to mediate transcellular delivery of viral proteins encompassing HLA class I-restricted epitopes. In this system, the recombinant protein TAT-NpFlu containing the transduction domain of Tat of human immunodeficiency virus type 1 fused to the amino acid region 301 to 498 of the nucleoprotein of influenza A virus is proven to sensitize different human cells to lysis by HLA-B27-restricted, Flu 383-391-specific CTL lines. The fusion protein is processed very effectively, since a comparable biological effect is obtained with an amount of protein between 1 and 2 orders of magnitude lower than that of the synthetic peptide. Interestingly, while part of TAT-NpFlu undergoes fast and productive cleavage, a large amount of it remains intact for up to 24 h. Confocal microscopy shows that TAT-NpFlu accumulates in the trans-Golgi network (TGN), where it starts to be detectable 1 h after transduction. Using TAT-NpFlu mutants and hybrid constructs, we demonstrate that enrichment in the TGN occurs only when the carboxy-terminal region of NpFlu (amino acids 400 to 498) is present. These data disclose an unconventional route for presentation of epitopes restricted for HLA class I molecules. PMID- 16306625 TI - Attenuation of simian immunodeficiency virus SIVmac239 infection by prophylactic immunization with dna and recombinant adenoviral vaccine vectors expressing Gag. AB - The prophylactic efficacy of DNA and replication-incompetent adenovirus serotype 5 (Ad5) vaccine vectors expressing simian immunodeficiency virus (SIV) Gag was examined in rhesus macaques using an SIVmac239 challenge. Cohorts of either Mamu A*01(+) or Mamu-A*01(-) macaques were immunized with a DNA prime-Ad5 boost regimen; for comparison, a third cohort consisting of Mamu-A*01(+) monkeys was immunized using the Ad5 vector alone for both prime and boost. All animals, along with unvaccinated control cohorts of Mamu-A*01(+) and Mamu-A*01(-) macaques, were challenged intrarectally with SIVmac239. Viral loads were measured in both peripheral and lymphoid compartments. Only the DNA prime-Ad5-boosted Mamu-A*01(+) cohort exhibited a notable reduction in peak plasma viral load (sevenfold) as well as in early set-point viral burdens in both plasma and lymphoid tissues (10 fold) relative to those observed in the control monkeys sharing the same Mamu A*01 allele. The degree of control in each animal correlated with the levels of Gag-specific immunity before virus challenge. However, virus control was short lived, and indications of viral escape were evident as early as 6 months postinfection. The implications of these results in vaccine design and clinical testing are discussed. PMID- 16306626 TI - Cytotoxic T-lymphocyte escape does not always explain the transient control of simian immunodeficiency virus SIVmac239 viremia in adenovirus-boosted and DNA primed Mamu-A*01-positive rhesus macaques. AB - Adenovirus 5 (Ad5) vectors show promise as human immunodeficiency virus vaccine candidates. Indian rhesus macaques vaccinated with Ad5-gag controlled simian human immunodeficiency virus SHIV89.6P viral replication in the absence of Env immunogens that might elicit humoral immunity. Here we immunized 15 macaques using either a homologous Ad5-gag/Ad5-gag (Ad5/Ad5) or a heterologous DNA-gag/Ad5 gag (DNA/Ad5) prime-boost regimen and challenged them with a high dose of simian immunodeficiency virus SIVmac239. Macaques vaccinated with the DNA/Ad5 regimen experienced a brief viral load nadir of less than 10,000 viral copies per ml blood plasma that was not seen in Mamu-A*01-negative DNA/Ad5 vaccinees, Mamu-A*01 positive Ad5/Ad5 vaccinees, or vaccine-naive controls. Interestingly, most of these animals were not durably protected from disease progression when challenged with SIVmac239. To investigate the reasons underlying this short-lived vaccine effect, we investigated breadth of the T-cell response, immunogenetic background, and viral escape from CD8+ lymphocytes that recognize immunodominant T-cell epitopes. We show that these animals do not mount unusually broad cellular immune response, nor do they express unusual major histocompatibility complex class I alleles. Viral recrudescence occurred in four of the five Mamu-A*01-positive vaccinated macaques. However, only a single animal in this group demonstrated viral escape in the immunodominant Gag181-189 CM9 response. These results suggest that viral "breakthrough" in vaccinated animals and viral escape are not inextricably linked and underscore the need for additional research into the mechanisms of vaccine failure. PMID- 16306627 TI - Restriction of human immunodeficiency virus type 1 by TRIM-CypA occurs with rapid kinetics and independently of cytoplasmic bodies, ubiquitin, and proteasome activity. AB - TRIM-CypA is an owl monkey-specific variant of the retrovirus restriction factor TRIM5alpha. Here, we exploit its modular domain organization and cyclosporine sensitivity to probe the kinetics and mechanism of TRIM5-mediated restriction. Time of addition/withdrawal experiments reveal that inhibition of incoming human immunodeficiency virus type 1 capsids by TRIM-CypA occurs within minutes of their delivery to the target cell cytoplasm. However, while TRIM-CypA restriction is partly dependent on a RING domain, restriction occurs independently of the ubiquitin/proteasome system. Moreover, tagged TRIM-CypA proteins can be fully active as restriction factors without forming cytoplasmic bodies. PMID- 16306628 TI - Identification of a functional envelope protein from the HERV-K family of human endogenous retroviruses. AB - Genome-wide screening of sequence databases for human endogenous retroviruses (HERVs) has led to the identification of 18 coding env genes, among which two-the syncytin genes-encode fusogenic ENV proteins possibly involved in placenta physiology. Here we show that a third ENV, originating from the most "recent" HERV-K(HML2) family, is functional. Immunofluorescence analysis of env-transduced cells demonstrates expression of the protein at the cell surface, and we show that the protein confers infectivity to simian immunodeficiency virus pseudotypes. Western blot analysis of the pseudotyped virions further discloses the expected specific cleavage of the ENV precursor protein. This functional ENV could play a role in the amplification--via infection of the germ line--of the HERV-K genomic copies, all the more as coding HERV-K gag and pol genes can similarly be found in the human genome, which could therefore generate infectious virions of a fully endogenous origin. PMID- 16306629 TI - Hepatitis C virus (HCV) and hepatitis B virus (HBV) can coinfect the same hepatocyte in the liver of patients with chronic HCV and occult HBV infection. AB - In this work, we have shown that hepatitis C virus (HCV) and hepatitis B virus (HBV) can coexist in the same hepatocyte using double fluorescent in situ hybridization in liver biopsy samples from patients with chronic HCV infection with occult HBV infection. Digital image analysis of hybridization signals showed that the HBV DNA levels in coinfected hepatocytes were lower than those in cells infected only with HBV. This finding supports the hypothesis of inhibition of HBV replication by HCV. Furthermore, HCV RNA levels were lower in coinfected cells than in cells infected only with HCV, suggesting that HBV may also inhibit HCV replication. PMID- 16306630 TI - A deubiquitinating activity is conserved in the large tegument protein of the herpesviridae. AB - The largest tegument protein of herpes simplex virus 1 (HSV-1), UL36, contains a novel deubiquitinating activity embedded in it. All members of the Herpesviridae contain a homologue of HSV-1 UL36, the N-terminal segments of which show perfect conservation of those residues implicated in catalysis. For murine cytomegalovirus and Epstein-Barr virus, chosen as representatives of the beta- and gammaherpesvirus subfamilies, respectively, we here show that the homologous modules indeed display deubiquitinating activity in vitro. The conservation of this activity throughout all subfamilies is indicative of an important, if not essential, function. PMID- 16306631 TI - Matrix-induced inhibition of membrane binding contributes to human immunodeficiency virus type 1 particle assembly defects in murine cells. AB - Defective human immunodeficiency virus type 1 (HIV-1) assembly in murine cells is accompanied by poor plasma membrane binding and proteolytic processing of the HIV 1 Gag precursor. Here, we show that such defects are induced by the propensity of the HIV-1 MA globular head to inhibit membrane binding and particle assembly, particularly at the low expression levels observed in murine cells. Simple additions to or deletion of the MA globular head can improve the yield of infectious virions from murine cells by >50-fold. Expression level and autoinhibition can be important confounding variables in studies of HIV-1 assembly and contribute to defects encountered in murine cells. PMID- 16306632 TI - MR imaging and MR spectroscopic imaging in the pre-treatment evaluation of prostate cancer. AB - Magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (1H MRSI) are emerging as the most sensitive tools for the non-invasive, anatomic and metabolic evaluation of prostate cancer. This article reviews the current applications of MRI and 1H MRSI in clinical practice and discusses the promise of these modalities for improving prostate cancer management. MRI demonstrates zonal anatomy with excellent contrast resolution and can reveal tumours in areas not routinely sampled on biopsy and not palpable on digital rectal examination. In addition, MR images allow assessment of local extent (including extracapsular extension and seminal vesicle invasion) and thus can assist in local staging while providing surgeons and radiation therapists with a visual road-map for treatment planning. The addition of 1H MRSI to MRI can improve prostate cancer detection and assessment of tumour volume; it also contributes indirectly to improved local staging. In addition, 1H MRSI metabolic and volumetric data correlate with pathological Gleason grade and thus may offer a non-invasive means to better predict prostate cancer aggressiveness. Combined MRI/1H MRSI is currently of greatest value for high-risk patients. With greater understanding of the relationship between spectroscopic data and tumour biology, it may become possible to use MRI/1H MRSI to achieve more precise stratification of patients in clinical trials, to monitor the progress of patients who select watchful waiting or minimally aggressive cancer therapies, and to guide and assess emerging local prostate cancer therapies. PMID- 16306633 TI - Advances in radiotherapy for prostate cancer. PMID- 16306634 TI - Thin section MRI in multidisciplinary pre-operative decision making for patients with rectal cancer. AB - High spatial resolution MRI of the rectum is an accurate method of staging rectal cancer. The technique requires attention to detail so that correct planes and scan parameters are used to obtain the best images. A detailed understanding of the pathological features of these tumours is required for image interpretation so that prognostic information beyond the basic T and N staging of the tumour can be obtained. Use of standardized criteria for reporting is reproducible in the multicentre setting and pre-operative multidisciplinary discussion of the MRI features increases the number of operations performed with tumour-free resection margins. PMID- 16306635 TI - Surgical management of rectal cancer: a multidisciplinary approach to technical and technological advances. PMID- 16306636 TI - Advances in chemoradiation therapy in rectal cancer: the impact of imaging. PMID- 16306638 TI - Intensity-modulated radiation therapy (IMRT): a clinical reality for cancer treatment, "any fool can understand this". The 2004 Silvanus Thompson Memorial Lecture. PMID- 16306639 TI - MR imaging in cervical cancer: seeing is believing. The 2004 Mackenzie Davidson Memorial Lecture. PMID- 16306640 TI - Pelvic malignancy: integrating form and function. AB - Despite the essential role morphological imaging plays in the management of patients with malignancy, anatomical techniques are limited in their ability to report on tumour biology and behaviour. It has therefore been necessary to develop imaging techniques that integrate form and function to probe the micro and molecular environments of cancers. The role of clinical functional and molecular magnetic resonance imaging is discussed with an emphasis on pelvic malignancy. It is argued that the radiological sciences need to take a lead in translating molecular and functional imaging techniques into man. Imaging in support of drug development is suggested as a focus for that development. PMID- 16306641 TI - A multidisciplinary approach to ovarian cancer at diagnosis. PMID- 16306642 TI - FPA-based infrared thermography as applied to the study of cutaneous perspiration and stimulated vascular response in humans. AB - This review gives an overview of focal plane array (FPA)-based infrared (IR) thermography as a powerful research method in the field of physiology and medicine. Comparison of the gained results with the data previously obtained by other authors with other research tools is given. Outer thermoregulatory manifestations displayed by the human organism subjected to whole-body heating (sauna bath) and physical loads (exercise bicycling) are quantitatively analysed. Some details of human body emotional sweating (psycho-physiological effect) are reported. Particular attention is paid to studying active sweat glands as individual objects. All experimental data were obtained with the help of a high sensitivity (0.03 degrees C) fast 128 x 128 InAs IR detector-based thermal imaging system operating in the short-wave spectral region (2.5 to 3 microm) and perfectly suiting medical purposes. It is shown that IR thermography makes it possible to overcome limitations inherent to contact measuring means that were traditionally used before in thermal studies. It is also shown that heterogeneous thermograms displayed by organisms with disturbed inner equilibrium can be quantitatively analysed in terms of statistical parameters of related surface temperature histograms, such as the mean temperature and the standard deviation of temperature (SDT). The increase and the decrease in SDT turned out to be typical of prolonged physical load and subsequent relaxation, and of external whole-body heating, respectively. Explanation of this result based on a hypothesis advanced within the context of the doctrine of human-organism evolution is given. Skin-temperature distribution function accompanying the relaxed organism in normality was found to closely resemble normal-distribution function. Symmetry break down and variation of the shape of this characteristic may serve as an indicator of homeostasis shift and can be used as a quantitative criterion for the latter. A new phenomenon, stable punctate hidrosis, is discovered and described. The term sweatology is introduced to refer to the discussed specific research area in biomedical science. PMID- 16306643 TI - Hyperspectral and multispectral bioluminescence optical tomography for small animal imaging. AB - For bioluminescence imaging studies in small animals, it is important to be able to accurately localize the three-dimensional (3D) distribution of the underlying bioluminescent source. The spectrum of light produced by the source that escapes the subject varies with the depth of the emission source because of the wavelength-dependence of the optical properties of tissue. Consequently, multispectral or hyperspectral data acquisition should help in the 3D localization of deep sources. In this paper, we describe a framework for fully 3D bioluminescence tomographic image acquisition and reconstruction that exploits spectral information. We describe regularized tomographic reconstruction techniques that use semi-infinite slab or FEM-based diffusion approximations of photon transport through turbid media. Singular value decomposition analysis was used for data dimensionality reduction and to illustrate the advantage of using hyperspectral rather than achromatic data. Simulation studies in an atlas-mouse geometry indicated that sub-millimeter resolution may be attainable given accurate knowledge of the optical properties of the animal. A fixed arrangement of mirrors and a single CCD camera were used for simultaneous acquisition of multispectral imaging data over most of the surface of the animal. Phantom studies conducted using this system demonstrated our ability to accurately localize deep point-like sources and show that a resolution of 1.5 to 2.2 mm for depths up to 6 mm can be achieved. We also include an in vivo study of a mouse with a brain tumour expressing firefly luciferase. Co-registration of the reconstructed 3D bioluminescent image with magnetic resonance images indicated good anatomical localization of the tumour. PMID- 16306644 TI - Targeting accuracy of an image guided gating system for stereotactic body radiotherapy. AB - Recently, a commercial system capable of x-ray image guided patient positioning and respiratory gated delivery has become available. Here we describe the operational principles of this system and investigate its geometric targeting accuracy under controlled conditions. The system tracks breathing via infrared (IR) detection of reflective markers located on the patient's abdomen. Localization kilovoltage (kV) x-rays are triggered from within the gated delivery window portion of the breathing trace and after positioning, the tumour will cross the linac isocentre during gated delivery. We tested geometric accuracy of this system by localizing and delivering gated fields to a moving phantom. Effects of phantom speed, gating window location, timing errors and phantom rotations on positioning and gating accuracy were investigated. The system delivered gated fields to both a moving and static phantom with equal accuracy. The position of the gating window affects accuracy only to the extent that an asymmetric breathing motion could affect dose distribution within its boundaries. Positioning errors were found to be less then 0.5 +/- 0.2 mm for phantom rotations up to 5 degrees. We found and corrected a synchronization error caused by a faulty x-ray duration setting and detected a 60 +/- 20 ms time delay in our linear accelerator. PMID- 16306645 TI - Robust optimization for intensity modulated radiation therapy treatment planning under uncertainty. AB - The recent development of intensity modulated radiation therapy (IMRT) allows the dose distribution to be tailored to match the tumour's shape and position, avoiding damage to healthy tissue to a greater extent than previously possible. Traditional treatment plans assume that the target structure remains in a fixed location throughout treatment. However, many studies have shown that because of organ motion, inconsistencies in patient positioning over the weeks of treatment, etc, the tumour location is not stationary. We present a probabilistic model for the IMRT inverse problem and show that it is identical to using robust optimization techniques, under certain assumptions. For a sample prostate case, our computational results show that this method is computationally feasible and promising-compared to traditional methods, our model has the potential to find treatment plans that are more adept at sparing healthy tissue while maintaining the prescribed dose to the target. PMID- 16306646 TI - Dosimetric characteristics of the Elekta Beam Modulator. AB - The dosimetric characteristics of a production pilot multi-leaf collimator (Elekta Beam Modulator, Elekta Oncology Systems, Crawley, UK) having a 4 mm leaf width (at isocentre) have been investigated. Characteristics explored included leaf bank set-up, penumbra width (80-20%) as a function of leaf position, leaf positioning reproducibility, interleaf leakage and leaf transmission. The penumbra values for leaf ends were measured to be between 4.2 and 4.8 mm for various large rectangular fields studied using Kodak X-omat V film at isocentre (1.5 cm deep). Similar films were taken with a standard 1 cm width multi-leaf collimator (MLC) and the penumbra for leaf ends was found to range from 4.3 to 5.2 mm. Other results showed that the rounded leaf tip provided tight control of the penumbra across the leaves' full range of travel. The positioning of the leaves was within a 0.5 mm range when approaching from the same direction. The maximum interleaf leakage was found to be 1.7% and the average leaf transmission less than 1.0%. No major differences were observed in leakage and transmission with changing gantry angle. PMID- 16306647 TI - Neutrons from fragmentation of light nuclei in tissue-like media: a study with the GEANT4 toolkit. AB - We study energy deposition by light nuclei in tissue-like media taking into account nuclear fragmentation reactions, in particular, production of secondary neutrons. The calculations are carried out within a Monte Carlo model for heavy ion therapy (MCHIT) based on the GEANT4 toolkit. Experimental data on depth-dose distributions for 135-400 A MeV (12)C and (18)O beams are described very well without any adjustment of the model parameters. This gives confidence in successful use of the GEANT4 toolkit for MC simulations of cancer therapy with beams of light nuclei. The energy deposition due to secondary neutrons produced by (12)C and (20)Ne beams in a (40-50 cm)(3) water phantom is estimated to be 1 2% of the total dose, that is only slightly above the neutron contribution (approximately 1%) induced by a 200 MeV proton beam. PMID- 16306648 TI - Optimal filter design for shielded and unshielded ambient noise reduction in fetal magnetocardiography. AB - The greatest impediment to extracting high-quality fetal signals from fetal magnetocardiography (fMCG) is environmental magnetic noise, which may have peak to-peak intensity comparable to fetal QRS amplitude. Being an unstructured Gaussian signal with large disturbances at specific frequencies, ambient field noise can be reduced with hardware-based approaches and/or with software algorithms that digitally filter magnetocardiographic recordings. At present, no systematic evaluation of filters' performances on shielded and unshielded fMCG is available. We designed high-pass and low-pass Chebychev II-type filters with zero phase and stable impulse response; the most commonly used band-pass filters were implemented combining high-pass and low-pass filters. The achieved ambient noise reduction in shielded and unshielded recordings was quantified, and the corresponding signal-to-noise ratio (SNR) and signal-to-distortion ratio (SDR) of the retrieved fetal signals was evaluated. The study regarded 66 fMCG datasets at different gestational ages (22-37 weeks). Since the spectral structures of shielded and unshielded magnetic noise were very similar, we concluded that the same filter setting might be applied to both conditions. Band-pass filters (1.0 100 Hz) and (2.0-100 Hz) provided the best combinations of fetal signal detection rates, SNR and SDR; however, the former should be preferred in the case of arrhythmic fetuses, which might present spectral components below 2 Hz. PMID- 16306649 TI - Magnetic images of the disintegration process of tablets in the human stomach by ac biosusceptometry. AB - Oral administration of solid dosage forms is usually preferred in drug therapy. Conventional imaging methods are essential tools to investigate the in vivo performance of these formulations. The non-invasive technique of ac biosusceptometry has been introduced as an alternative in studies focusing on gastrointestinal motility and, more recently, to evaluate the behaviour of magnetic tablets in vivo. The aim of this work was to employ a multisensor ac biosusceptometer system to obtain magnetic images of disintegration of tablets in vitro and in the human stomach. The results showed that the transition between the magnetic marker and the magnetic tracer characterized the onset of disintegration (t(50)) and occurred in a short time interval (1.1 +/- 0.4 min). The multisensor ac biosusceptometer was reliable to monitor and analyse the in vivo performance of magnetic tablets showing accuracy to quantify disintegration through the magnetic images and to characterize the profile of this process. PMID- 16306650 TI - Implementation and analysis of relief patterns of the surface of benign and malignant lesions of the skin by microtopography. AB - The objective of this study was to be able to distinguish between healthy skin tissue and malignant ones, furthermore determining a unique pattern of roughness for each skin lesion by microtopographic analysis of the skin surface of Mexican patients during the period from April to October 2002. The standard technique used in this study for the diagnosis of skin cancer and the comparison of the results was the haematoxylin-eosin histopathological technique. Latex impressions were taken from skin lesions as well as from the healthy skin of each patient to serve as control samples. These impressions were analysed by the MICROTOP.03.MFC microtopographic system inspection. It was observed that when the tumour becomes rougher, more malign will be the lesion. On average, the melanoma present an increase of roughness of 67% compared to healthy skin, obtaining a roughness relation of 1:2.54. The percentage decreases to 49% (49%, 1:60) in the case of basal cell carcinoma and to 40% in pre-malignant lesions such as melanocytic nevus (40%, 1:150). In benign lesions such as the seborrhoea keratosis only a small increase in roughness was noted (4%, 1:0.72). Microtopographic inspection of the skin surface can be considered as a complementary diagnostic technique for skin cancer. PMID- 16306651 TI - Three-dimensional simulation of ultrasound propagation through trabecular bone structures measured by synchrotron microtomography. AB - Three-dimensional numerical simulations of ultrasound transmission were performed through 31 trabecular bone samples measured by synchrotron microtomography. The synchrotron microtomography provided high resolution 3D mappings of bone structures, which were used as the input geometry in the simulation software developed in our laboratory. While absorption (i.e. the absorption of ultrasound through dissipative mechanisms) was not taken into account in the algorithm, the simulations reproduced major phenomena observed in real through-transmission experiments in trabecular bone. The simulated attenuation (i.e. the decrease of the transmitted ultrasonic energy) varies linearly with frequency in the MHz frequency range. Both the speed of sound (SOS) and the slope of the normalized frequency-dependent attenuation (nBUA) increase with the bone volume fraction. Twenty-five out of the thirty-one samples exhibited negative velocity dispersion. One sample was rotated to align the main orientation of the trabecular structure with the direction of ultrasonic propagation, leading to the observation of a fast and a slow wave. Coupling numerical simulation with real bone architecture therefore provides a powerful tool to investigate the physics of ultrasound propagation in trabecular structures. As an illustration, comparison between results obtained on bone modelled either as a fluid or a solid structure suggested the major role of mode conversion of the incident acoustic wave to shear waves in bone to explain the large contribution of scattering to the overall attenuation. PMID- 16306652 TI - Spectral image reconstruction for transcranial ultrasound measurement. AB - An approach aimed at improved ultrasound resolution and signal strength through highly attenuating media is presented. The method delivers a series of multiple cycle bursts in order to construct a discrete spectral (frequency domain) response in one dimension. Cross-correlation of this ultrasound A-mode response with its transmitted signal results in time-localized peaks that correspond to scattering locations. The approach is particularly relevant to the problem of transcranial ultrasound imaging, as it combines numerous smaller signals into a single signal whose net power may exceed that which could be achieved using a single burst. Tests are performed with human skull fragments and nylon-wire targets embedded in a tissue phantom. Skulls are oriented to produce both lateral and shear modes of transcranial propagation. A total of nine locations distributed over three ex vivo human skull samples are studied. Compared with pulsed and chirped signals, results indicate more localized peaks when using the multi-cycle approach, with more accurate positioning when combined with the transcranial shear mode. PMID- 16306653 TI - Monte Carlo simulations of increased/decreased scattering inclusions inside a turbid slab. AB - We analyse the effect on scattered photons of anomalous optical inclusions in a turbid slab with otherwise uniform properties. Our motivation for doing so is that inclusions affect scattering contrast used to quantify optical properties found from transmitted light intensity measured in transillumination experiments. The analysis is based on a lattice random walk formalism which takes into account effects of both positive and negative deviations of the scattering coefficient from that of the bulk. Our simulations indicate the existence of a qualitative difference between the effects of these two types of perturbations. In the case of positive perturbations the time delay is found to be proportional to the square of the size of the inclusion while for negative perturbations the time delay is a linear function of its volume. PMID- 16306654 TI - A differentiable Shepp-Logan phantom and its applications in exact cone-beam CT. AB - Recently, several exact cone-beam reconstruction algorithms, such as the generalized filtered-backprojection (FBP) and backprojection-filtration (BPF) methods, have been developed to solve the long object problem. Although the well known 3D Shepp-Logan phantom (SLP) is often used to validate these algorithms, it is deficient due to the discontinuity of the SLP. In this paper, we first construct a differentiable polynomial function to approximate the unit rectangular function on [-1, 1]. Then, we use this function to obtain a differentiable ellipsoid phantom, whose x-ray transform is differentiable for any smooth scanning trajectory. Finally, we propose a differentiable Shepp-Logan phantom (DSLP) for numerical simulation of the exact cone-beam CT algorithms. Our numerical simulation shows that the reconstructed DSLP has a better image quality than the reconstructed SLP, and is complementary to the traditional SLP for evaluation of the exact cone-beam CT algorithms. PMID- 16306655 TI - Tissue-mimicking agar/gelatin materials for use in heterogeneous elastography phantoms. AB - Five 9 cm x 9 cm x 9 cm phantoms, each with a 2-cm-diameter cylindrical inclusion, were produced with various dry-weight concentrations of agar and gelatin. Elastic contrasts ranged from 1.5 to 4.6, and values of the storage modulus (real part of the complex Young's modulus) were all in the soft tissue range. Additives assured immunity from bacterial invasion and can produce tissue mimicking ultrasound and NMR properties. Monitoring of strain ratios over a 7 to 10 month period indicated that the mechanical properties of the phantoms were stable, allowing about 1 month for the phantom to reach chemical equilibrium. The only dependable method for determining the storage moduli of the inclusions is to make measurements on samples excised from the phantoms. If it is desired to produce and accurately characterize a phantom with small inclusions with other shapes, such as an array of small spheres, an auxiliary phantom with the geometry of the cylindrical inclusion phantoms or the equivalent should be made at the same time using the same materials. The elastic contrast can then be determined using samples excised from the auxiliary phantom. A small increase of about 10% in volume of the cylindrical inclusions occurred-a tolerable increase. Interestingly, the smallest increase (about 5%) occurred in the phantom with the largest elastic contrast. PMID- 16306656 TI - High-precision dosimetry for radiotherapy using the optically stimulated luminescence technique and thin Al2O3:C dosimeters. AB - The potential of using the optically stimulated luminescence (OSL) technique with aluminium oxide (Al(2)O(3):C) dosimeters for a precise and accurate estimation of absorbed doses delivered by high-energy photon beams was investigated. This study demonstrates the high reproducibility of the OSL measurements and presents a preliminary determination of the depth-dose curve in water for a 6 MV photon beam from a linear accelerator. The uncertainty of a single OSL measurement, estimated from the variance of a large sample of dosimeters irradiated with the same dose, was 0.7%. In the depth-dose curve obtained using the OSL technique, the difference between the measured and expected doses was < or =0.7% for depths between 1.5 and 10 cm, and 1.1% for a depth of 15 cm. The readout procedure includes a normalization of the response of the dosimeter with respect to a reference dose in order to eliminate variations in the dosimeter mass, dosimeter sensitivity, and the reader's sensitivity. This may be relevant for quality assurance programmes, since it simplifies the requirements in terms of personnel training to achieve the precision and accuracy necessary for radiotherapy applications. We concluded that the OSL technique has the potential to be reliably incorporated in quality assurance programmes and dose verification. PMID- 16306657 TI - On the existence of low-energy photons (<150 keV) in the unflattened x-ray beam from an ordinary radiotherapeutic target in a medical linear accelerator. AB - Low-energy photons (<150 keV) are essential for obtaining high quality x-ray radiographs. These photons are usually produced in the accelerator target, but are effectively absorbed by the flattening filter and, at least partially, by the target itself. Experimental proof is presented for the existence of low-energy photons in the unflattened x-ray beam produced by a 6 MeV electron beam normally incident on the thinner of the two existing ports of the all-Cu radiotherapeutic target of a Clinac 18 (Varian Associates) linear accelerator. A number of one shot absorption measurements were carried out with 12 foils of Pb absorbers with thicknesses varying from 0.25 to 3 mm in steps of 0.25 mm arranged symmetrically around the central axis on a 7.2 cm radius circumference. A Kodak ECL film-screen cassette combination was used as a detector in the absorption measurements, in which optical density was measured as a function of the thickness of the Pb absorbers. Two sets of absorption measurements were carried out: the first one with the Clinac 18 6 MV unflattened beam and the second one with the Clinac 600C 6 MV therapeutic counterpart beam. There is a striking difference between the two sets: the optical density versus Pb-absorber thickness curve shows a sharp increase in optical density at small absorber thicknesses in the case of the unflattened 6 MV x-ray beam as compared with a gently sloping dependence in the case of the 6 MV therapeutic beam. A semi-quantitative assessment of the low energy photon contribution to the whole optical density/contrast is presented. A 0.85 mm thick Pb absorber intercepting the 6 MV unflattened x-ray beam eliminates almost totally the sharp peak in the optical density curve at small Pb-absorber thicknesses. This constitutes additional evidence for the existence of low-energy photons (<150 keV) in the unflattened 6 MV beam from the Cu therapeutic target. PMID- 16306658 TI - A dose verification method using a monitor unit matrix for dynamic IMRT on Varian linear accelerators. AB - Dosimetry verification is an important step during intensity modulated radiotherapy treatment (IMRT). The verification is usually conducted with measurements and independent dose calculations. However, currently available independent dose calculation methods were developed for step-and-shoot beam delivery methods, and their uses for dynamic multi-leaf collimator (MLC) delivery methods are not efficient. In this study, a dose calculation method was developed to perform independent dose verifications for a dynamic MLC-based IMRT technique for Varian linear accelerators. This method extracts the machine delivery parameters from the dynamic MLC (DMLC) files generated by the IMRT treatment planning system. Based on the parameters a monitor unit (MU) matrix was separately calculated as two terms: direct exposure from the open MLC field and leakage contributions, where the leaf-end leakage contribution becomes more important in higher dose gradient regions. The MU matrix was used to compute the primary dose and the scattered dose with a modified Clarkson technique. The doses computed using the method were compared with both measurement and treatment planning for 14 and 25 plans respectively. An average of less than 2% agreement was observed and the standard deviation was about 1.9%. PMID- 16306659 TI - An examination of the number of required apertures for step-and-shoot IMRT. AB - We have examined the degree to which step-and-shoot IMRT treatment plans can be simplified (using a small number of apertures) without sacrificing the dosimetric quality of the plans. A key element of this study was the use of direct aperture optimization (DAO), an inverse planning technique where all of the multi-leaf collimator constraints are incorporated into the optimization. For seven cases (1 phantom, 1 prostate, 3 head-and-neck and 2 lung), DAO was used to perform a series of optimizations where the number of apertures per beam direction varied from 1 to 15. In this work, we attempt to provide general guidelines for how many apertures per beam direction are sufficient for various clinical cases using DAO. Analysis of the optimized treatment plans reveals that for most cases, only modest improvements in the objective function and the corresponding DVHs are seen beyond 5 apertures per beam direction. However, for more complex cases, some dosimetric gain can be achieved by increasing the number of apertures per beam direction beyond 5. Even in these cases, however, only modest improvements are observed beyond 9 apertures per beam direction. In our clinical experience, 38 out of the first 40 patients treated using IMRT plans produced using DAO were treated with 9 or fewer apertures per beam direction. The results indicate that many step-and-shoot IMRT treatment plans delivered today are more complex than necessary and can be simplified without sacrificing plan quality. PMID- 16306660 TI - Effects of bone- and air-tissue inhomogeneities on the dose distributions of the Leksell Gamma Knife calculated with PENELOPE. AB - Monte Carlo simulation with PENELOPE (version 2003) is applied to calculate Leksell Gamma Knife dose distributions for heterogeneous phantoms. The usual spherical water phantom is modified with a spherical bone shell simulating the skull and an air-filled cube simulating the frontal or maxillary sinuses. Different simulations of the 201 source configuration of the Gamma Knife have been carried out with a simplified model of the geometry of the source channel of the Gamma Knife recently tested for both single source and multisource configurations. The dose distributions determined for heterogeneous phantoms including the bone- and/or air-tissue interfaces show non-negligible differences with respect to those calculated for a homogeneous one, mainly when the Gamma Knife isocentre approaches the separation surfaces. Our findings confirm an important underdosage (approximately 10%) nearby the air-tissue interface, in accordance with previous results obtained with the PENELOPE code with a procedure different from ours. On the other hand, the presence of the spherical shell simulating the skull produces a few per cent underdosage at the isocentre wherever it is situated. PMID- 16306661 TI - Performance of three-photon PET imaging: Monte Carlo simulations. AB - We have recently introduced the idea of making use of three-photon positron annihilations in positron emission tomography. In this paper, the basic characteristics of the three-gamma imaging in PET are studied by means of Monte Carlo simulations and analytical computations. Two typical configurations of human and small animal scanners are considered. Three-photon imaging requires high-energy resolution detectors. Parameters currently attainable by CdZnTe semiconductor detectors, the technology of choice for the future development of radiation imaging, are assumed. Spatial resolution is calculated as a function of detector energy resolution and size, position in the field of view, scanner size and the energies of the three-gamma annihilation photons. Possible ways to improve the spatial resolution obtained for nominal parameters, 1.5 cm and 3.2 mm FWHM for human and small animal scanners, respectively, are indicated. Counting rates of true and random three-photon events for typical human and small animal scanning configurations are assessed. A simple formula for minimum size of lesions detectable in the three-gamma based images is derived. Depending on the contrast and total number of registered counts, lesions of a few mm size for human and sub mm for small animal scanners can be detected. PMID- 16306662 TI - A count-rate model for PET scanners using pixelated Anger-logic detectors with different scintillators. AB - A high count-rate simulation (HCRSim) model has been developed so that all results are derived from fundamental physics principles. Originally developed to study the behaviour of continuous sodium iodide (NaI(Tl)) detectors, this model is now applied to PET scanners based on pixelated Anger-logic detectors using lanthanum bromide (LaBr(3)), gadolinium orthosilicate (GSO) and lutetium orthosilicate (LSO) scintillators. This simulation has been used to study the effect on scanner deadtime and pulse pileup at high activity levels due to the scintillator stopping power (mu), decay time (tau) and energy resolution. Simulations were performed for a uniform 20 cm diameter x 70 cm long cylinder (NEMA NU2-2001 standard) in a whole-body scanner with an 85 cm ring diameter and a 25 cm axial field-of-view. Our results for these whole-body scanners demonstrate the potential of a pixelated Anger-logic detector and the relationship of its performance with the scanner NEC rate. Faster signal decay and short coincidence timing window lead to a reduction in deadtime and randoms fraction in the LaBr(3) and LSO scanners compared to GSO. The excellent energy resolution of LaBr(3) leads to the lowest scatter fraction for all scanners and helps compensate for reduced sensitivity compared to the GSO and LSO scanners, leading to the highest NEC values at high activity concentrations. The LSO scanner has the highest sensitivity of all the scanner designs investigated here, therefore leading to the highest peak NEC value but at a lower activity concentration than that of LaBr(3). PMID- 16306663 TI - Optimization of multi-slice helical respiration-correlated CT: the effects of table speed and rotation time. AB - While respiration-correlated CT is gaining acceptance in clinical radiotherapy, the effect of scanning parameters on the image quality has yet to be addressed. The intent of this study was to characterize the effects of gantry rotation and table speed on various image quality characteristics in multi-slice, helical, retrospectively-gated CT images. Images of stationary and moving phantoms were obtained in helical mode on a 20-slice CT scanner. Motion was generated by a computer-controlled platform capable of moving simultaneously in two dimensions. Motion was monitored using a pressure gauge inserted inside an adjustable belt. Selected scans were retrospectively gated into ten phases based on the monitored motion. Gantry rotation speeds of 0.5 s and 1.0 s were evaluated with pitches ranging from 0.1 to 0.45. Several parameters, including calculated object volumes, trajectory (movement from peak to trough), deformation (actual volume divided by volume created with the maximum diameter of contoured object) and z axis resolution, were used to characterize image quality. These studies indicate that for objects in the peak phase of a movement pattern that simulates breathing, retrospectively gated scans using fast gantry rotation speeds produce volume, trajectory, deformation and z-axis resolution results comparable with those of a stationary object. PMID- 16306664 TI - Dynamic flat panel detector versus image intensifier in cardiac imaging: dose and image quality. AB - The practical aspects of the dosimetric and imaging performance of a digital x ray system for cardiology procedures were evaluated. The system was configured with an image intensifier (II) and later upgraded to a dynamic flat panel detector (FD). Entrance surface air kerma (ESAK) to phantoms of 16, 20, 24 and 28 cm of polymethyl methacrylate (PMMA) and the image quality of a test object were measured. Images were evaluated directly on the monitor and with numerical methods (noise and signal-to-noise ratio). Information contained in the DICOM header for dosimetry audit purposes was also tested. ESAK values per frame (or kerma rate) for the most commonly used cine and fluoroscopy modes for different PMMA thicknesses and for field sizes of 17 and 23 cm for II, and 20 and 25 cm for FD, produced similar results in the evaluated system with both technologies, ranging between 19 and 589 microGy/frame (cine) and 5 and 95 mGy min(-1) (fluoroscopy). Image quality for these dose settings was better for the FD version. The 'study dosimetric report' is comprehensive, and its numerical content is sufficiently accurate. There is potential in the future to set those systems with dynamic FD to lower doses than are possible in the current II versions, especially for digital cine runs, or to benefit from improved image quality. PMID- 16306665 TI - Evaluation of a software package for automated quality assessment of contrast detail images--comparison with subjective visual assessment. AB - Contrast detail analysis is commonly used to assess image quality (IQ) associated with diagnostic imaging systems. Applications include routine assessment of equipment performance and optimization studies. Most frequently, the evaluation of contrast detail images involves human observers visually detecting the threshold contrast detail combinations in the image. However, the subjective nature of human perception and the variations in the decision threshold pose limits to the minimum image quality variations detectable with reliability. Objective methods of assessment of image quality such as automated scoring have the potential to overcome the above limitations. A software package (CDRAD analyser) developed for automated scoring of images produced with the CDRAD test object was evaluated. Its performance to assess absolute and relative IQ was compared with that of an average observer. Results show that the software does not mimic the absolute performance of the average observer. The software proved more sensitive and was able to detect smaller low-contrast variations. The observer's performance was superior to the software's in the detection of smaller details. Both scoring methods showed frequent agreement in the detection of image quality variations resulting from changes in kVp and KERMA(detector), which indicates the potential to use the software CDRAD analyser for assessment of relative IQ. PMID- 16306666 TI - Quantitative fluorescence microscopy of macromolecules in gel and biological tissue. AB - Quantitative fluorescence microscopy provides valuable insight into drug delivery and pharmacokinetics. The technique is based on analysis of statistical fluctuations in fluorescence that arises as fluorophores pass through a small volume illuminated by a focused laser beam, and has been applied to measure particle motion and binding interactions in solutions, on surfaces and inside the cells. We examined the use of fluorescence correlation spectroscopy combined with a microscope (FCSM) to assess the transport of fluorescent beads and macromolecules in aqueous solutions, gels and living biological tissue. Obstructed diffusion of fluorescent beads in gels of various densities was tested to get a sensible estimate of diffusion in the interstitial tissue matrix consistent with previous reports. Fluorescently labelled liposomes as an artificial drug or gene carrying vehicles were used for pharmacokinetic tests of drug delivery in living tissue. The results indicate that FCS is an accurate and valuable tool for measuring the physical properties of gene vectors in vitro and for characterizing interactions with tissue in vivo. PMID- 16306667 TI - A reinterpretation of the phase transitions in Na2CO3. AB - Based on the structural data of phases alpha (hexagonal; 756-972 K), beta (monoclinic; 605-751 K), gamma (incommensurate, monoclinic; 295 K) and delta (lock-in, monoclinic; 110 K) of sodium carbonate, Na2CO3, we could draw a parallel between the phase transitions and the evolution of the second coordination sphere of the C atoms. The temperature-dependent structures observed in the beta phase are reproduced in the incommensurate gamma phase as a modulation wave, which relates to the content of the symmetrically equivalent {110} lattice planes in the alpha phase. By decreasing the temperature, the phase transitions are associated with a stepwise increase in the number of Na ions participating in the second coordination sphere of the C atoms. Over the full temperature range, this number increases from 3 to 7. The C-O distances and the mobility of the O atoms depends on the number of Na ions in the vicinity of the C atoms. PMID- 16306668 TI - X-ray diffraction study for one-dimensional ionic conductors K(x)(Ga(1 - y)Al(y))(2 + x)Ti(2 - x)O(7) (x approximately = 0.14, y approximately = 0.10, 0.23, 0.39). AB - The structures of one-dimensional ionic conductors K(x)(Ga(1 - y)Al(y))(2 + x)Ti(2 - x)O(7) (x approximately 0.14, y approximately 0.10, 0.23, 0.39) were refined by single-crystal X-ray diffraction. A one-dimensional tunnel-like space with a large cross section is formed by the linkage of coordination polyhedra of the metal and oxygen ions; K ions are distributed in the tunnel. Significant differences were seen in structures with different Al content; these differences could be explained by considerations in crystal chemistry. The probability density functions (PDFs) of the K ion were obtained using up to fourth-order terms of the atomic displacement parameters. The joint PDFs for the K ion have clarified that a K conduction path deviates slightly from the central axis of the tunnel in all the samples. In contrast with the usual one-dimensional ionic conductors, no distinct bottleneck effect was observed from the joint-PDFs and one-particle potentials. PMID- 16306669 TI - Tilt and acoustic instabilities in ABX4, A2BX4 and ABX3 perovskite structure types: their role in the incommensurate phases of the organic-inorganic perovskites. AB - An examination of the tilt modes and other low-frequency modes is made for an isolated, untilted perovskite layer, which maps very simply to the ABX4 perovskites. A sheet of pure tilts exists at the Brillouin Zone boundary at {xi, (1/2), zeta}. An instability is also found at all wavevectors which can be described as continuously varying from pure tilts to pure layer displacements as a function of the wavevector. Analysis is extended by considering the stacking of layers in the I-centered A2BX4-layer perovskites. The effect of freezing in the commensurate tilt required to generate the Cmca tilt system, pertinent to the modulated phases of propylammonium salts, is examined. The zero-frequency modes are restricted to two planes in the Brillouin Zone. All of the observed wavevectors associated with modulated phases, and the commensurately tilted propylammonium tetrachlorocadmate, are consistent with this calculation. The effect of full three-dimensional connectivity is briefly reviewed for the true ABX3 perovskites. While pure tilt incommensurates appear to be hypothetically possible, they do not appear to have been observed to date. PMID- 16306670 TI - Second-degree twinning and dynamic disorder in the crystal structure of deca dodecasil 3R. AB - The structure of deca-dodecasil 3R (DD-3R), Si120O240, a very well suited material for the synthesis of inorganic/organic composites structured on a nanometer level, has been investigated in detail. So far, a highly complicated twinning has hampered its structure description at a desirable level of accuracy. This twinning has now been resolved and a new structure determination is presented. Structure refinement in the R?bar 3 space group revealed a large, unusually shaped atomic displacement ellipsoid for oxygen-bridging units (tetrahedra), bridging Si-O bonds shorter than expected and the linear Si-O-Si' bond angle dictated by special positions at a threefold axis. A structure model based on a statistically disordered bridging O atom improved the accuracy of the Si-O bonds of interest, but provided unacceptable O-O contacts. To solve this dilemma, ab initio NVT molecular dynamics calculations were performed to study the possible configurations. Wavelet analysis of the time variations of selected Si-O distances pointed to a synchronous shift of the whole building units (tetrahedra). Low-frequency features of the calculated phonon density of states agree well with the published INS (inelastic neutron scattering) spectra of several silica polymorphs, indicating that the nature of the disorder in DD-3R is dynamic rather than static. PMID- 16306671 TI - Geometrical parameterization of the crystal chemistry of P6(3)/m apatites: comparison with experimental data and ab initio results. AB - Experimental structure refinements and ab initio simulation results for 18 published, fully ordered P6(3)/m (A;{?rm I}_4)(A;{?rm II}_6)(BO4)6X2 apatite end member compositions have been analyzed in terms of a geometric crystal-chemical model that allows the prediction of unit-cell parameters (a and c) and all atom coordinates. To an accuracy of +/- 0.025 A, the magnitude of c was reproduced from crystal-chemical parameters characterizing chains of ...-A(II)-O3-B-O3-A(II) ... atoms, whereas that of a was determined from those describing (A(I)O6)-(BO4) polyhedral arrangements. The c/a ratio could be predicted to +/-0.2% using multi variable functions based on geometric crystal-chemical model predictions, but could not be ascribed to the adjustment of a single crystal-chemical parameter. The correlations observed between algebraically independent crystal-chemical parameters representing the main observed polyhedral distortions reveal them as the minimum-energy solution to accommodate misfit components within this flexible structure type. For materials with given composition, good agreement (within +/- 0.5-2.0%) of ab initio crystal-chemical parameters was observed with only those from single-crystal refinements with R 4.0% was not as good, while the scatter with those from Rietveld refinements was considerable. Accordingly, ab initio cell data, atomic coordinates and crystal-chemical parameters were reported here for the following compositions awaiting experimental work: (Zn,Hg)10(PO4)6(Cl,F)2, (Ca,Cd)10(VO4)6Cl2 and (Ca,Pb,Cd)10(CrO4)6Cl2. PMID- 16306672 TI - Incommensurately modulated ordering of tetrahedral chains in Ca2Fe2O5 at elevated temperatures. AB - The basic building units of brownmillerite-type A2B2O5 structures are perovskite like layers of corner-sharing BO6 octahedra and zweier single chains of BO4 tetrahedra. A three-dimensional framework is formed by alternate stacking of octahedral layers and sheets of tetrahedral chains. The compound Ca2Fe2O5 is known to have Pnma symmetry at ambient conditions. The space group Imma was reported to be evident above 963 K. New high-temperature single-crystal X-ray diffraction experiments at 1100 K revealed that Ca2Fe2O5 forms an incommensurately modulated structure adopting the superspace group Imma(00gamma)s00, with gamma = 0.588 (2). The modulation affects the sequence of the enantiomorphic (right- and left-handed) oriented tetrahedral chains within the layer, breaking the lattice periodicity along c. This ordering can be modelled with crenel occupation modulation functions for the tetrahedrally coordinated Fe, as well as for the O atom interconnecting the tetrahedra. PMID- 16306673 TI - Structures of Na9[SO4]4X.2H2O2, where X = Cl or Br, in which the halide anions orchestrate extended orientation sequences of H2O2 solvate molecules. AB - Detailed structures of nonasodium tetrakis(sulfate) chloride diperhydrate, Na9[SO4]4Cl.2H2O2, and its novel bromide analogue are compared. Hydrogen peroxide could not be resolved in a previously reported Na9[SO4]4Cl.2H2O2 substructure. However, on lowering the symmetry to P4/n, and using reflection data based on full unit-cells, the H2O2 solvate can be clearly seen. Although H2O2 molecules are not directly bonded to the halide anions, they exert considerable influence on the eight sodium cations that constitute each halide's coordination shell so that H2O2 ordering can be linked to halide dimensions. PMID- 16306674 TI - Structural characterization of Cd3(O3PC2H4CO2)2.2H2O from in-house X-ray powder data and NMR. AB - The title compound poly[[bis(mu-2-carboxylatoethylphosphonato)cadmium] dihydrate], Cd3(O3PC2H4CO2)2.2H2O, was prepared by a hydrothermal reaction and its crystal structure determined from in-house powder data. The structure was solved in both P2(1)/c and P2(1) space groups. The refinement converged with Rp = 0.1046, R(wp) = 0.1378 and Rf = 0.0763 in P2(1)/c. However, the solid-state NMR data could not be explained. The structure was then solved in P2(1) and the refinement converged with Rp = 0.0750, Rwp = 0.1022 and R(f) = 0.0409 and satisfied the NMR requirements. PMID- 16306675 TI - Three modulation patterns in four related [M(H2O)2(15-crown-5)](NO3)2 structures. AB - The structures of [M(H2O)2(15-crown-5)](NO3)2, M = Cu, Zn, Mg and Co, and 15 crown-5 = 1,4,7,10,13-pentaoxacyclopentadecane, have been redetermined at 294 and 90 K. The four structures, and a second form of the Cu structure, have been reported in the literature, but are all incorrect in some significant way. The structures, which all have at least two independent formula units (i.e. Z' > or = 2), are related; each water ligand is hydrogen-bonded to two nitrate anions, while each nitrate anion is hydrogen bonded to the water ligands of two cations. In the tetragonal Co structure the hydrogen-bond pattern is three-dimensional; in the monoclinic Cu, Zn and Mg structures the hydrogen-bond patterns are two dimensional. In the isostructural Zn and Mg structures Z' = 3, while in the Cu structure Z' = 5. The Cu, Zn and Mg structures are modulated variants of a basic structure, which was reported for Cu but which probably does not exist. The conformations of the 11 independent cations are remarkably similar; they all have approximate twofold symmetry and so exist as conformational enantiomers. The most important modulation is imperfect enantiomeric alternation of the cations along the longest cell axis; the independent cations are related by very good pseudotranslation and pseudoinversion operations. The diffraction patterns for all four structures have classes of weak, even very weak, reflections. PMID- 16306676 TI - The unusual phases of anhydrous and hydrated pinacol. AB - The structure of highly twinned pinacol (2,3-dimethyl-2,3-butanediol) monohydrate, the existence of which has been known since 1922, has been determined, and the structures of anhydrous pinacol and its two other known hydrates have been reinvestigated. All the phases are unusual. The anhydrous phase is exceptional among molecular crystals in having molecules located on three different symmetry sites (1, ?overline 1 and 2). A hexagonal form of pinacol originally described as a second polymorph has been shown to be a solvate of uncertain composition that is very loosely packed. Pinacol hexahydrate, which was originally reported as tetragonal and highly disordered, appears to be described better as having an orthorhombic structure that is both disordered and twinned; the diffraction pattern at 90 K shows structured diffuse scattering that suggests short-range correlations of disordered molecules. The occurrence of this unusual set of structures is attributed to the combination of the hydrogen bonding requirements of the pinacol molecule with its small size and limited conformational flexibility. PMID- 16306677 TI - Isostructural phase transition in m-carboxyphenylammonium monohydrogenphosphite. AB - Crystals of m-carboxyphenylammonium monohydrogenphosphite, C7H8NO2+.H2PO3- (m CPAMP), space group P2(1)/c, grown from aqueous solution undergo a reversible first-order single-crystal phase transition at Tc = 246 (2) K with a hysteresis of 3.6 K. The thermal behaviour of the sample was characterized by differential scanning calorimetry (DSC) experiments. Variations of the unit-cell parameters versus temperature between 100 and 320 K are reported. The transition from the higher-temperature phase (HTP) to the lower-temperature phase (LTP) is characterized by a unit-cell volume contraction of 1.77%. The average structure and unit-cell packing of m-CPAMP at lower temperature (100 K) are reported from accurate X-ray data sets and compared with those of the higher-temperature phase (293 K) in order to investigate the mechanism of the phase transition. The reciprocal lattice reconstruction showed a few very weak satellite reflections which will be discussed in a forthcoming paper. PMID- 16306678 TI - Structure and intermolecular interactions of glipizide from laboratory X-ray powder diffraction. AB - The crystal structure of glipizide, used as a major treatment of type-2 diabetes, has been determined ab initio using variable-temperature laboratory X-ray powder diffraction combined with a direct-space Monte Carlo/simulated annealing methodology. The strengths of the intermolecular interactions (van der Waals, pi pi stacking, hydrogen bonding and steric interlock) were quantitatively estimated using the thermal expansion data, which were collected in the same set of experiments as those used to determine the structure. PMID- 16306679 TI - Pressure-induced polymorphism in cyclopropylamine. AB - The crystal structure of cyclopropylamine at 1.2 GPa has been determined by X-ray diffraction methods. The structure of this phase is orthorhombic, space group Pbca and the unit-cell dimensions are a = 5.0741 (10), b = 9.7594 (10) and c = 13.305 (2) A. Only one of the two H atoms of the amino group actively participates in the formation of the hydrogen-bonded chains, C(2) in graph-set notation, which lie parallel to the crystallographic a axis. Additionally, the topology of the crystal packing is studied using both Voronoi-Dirichlet polyhedra and Hirshfeld surface analyses for the low-temperature and the high-pressure structures of cyclopropylamine and the results are compared. PMID- 16306680 TI - Variable-temperature neutron diffraction studies of the short, strong hydrogen bonds in the crystal structure of pyridine-3,5-dicarboxylic acid. AB - Pyridine-3,5-dicarboxylic acid has been studied by single-crystal neutron diffraction at 15 and 296 K. Pyridine-3,5-dicarboxylic acid, in which the carboxylic acid protons have been replaced by deuterons, has also been studied at 15, 150 and 296 K. The protonated structure contains a short N...H...O hydrogen bond [N...O 2.523 (2) Angstrom at 15 K]. Temperature-dependent proton migration occurs where the N--H distance in the hydrogen bond changes from 1.213 (4) Angstrom at 15 K to 1.308 (6) Angstrom at 300 K. In the deuterated structure the overall hydrogen-bond length increased [N...O 2.538 (3) Angstrom at 15 K] and the magnitude of the migration increased so that the N--D distance changes from 1.151 (3) Angstrom at 15 K to 1.457 (4) Angstrom at 300 K. PMID- 16306681 TI - Recent advances in structural research on ether lipids from archaea including comparative and physiological aspects. AB - A great number of novel and unique chemical structures of archaeal polar lipids have been reported. Since 1993, when those lipids were reviewed in several review articles, a variety of core lipids and lipids with unique polar groups have been reported successively. We summarize new lipid structures from archaea elucidated after 1993. In addition to lipids from intact archaeal cells, more diverse structures of archaea-related lipids found in environmental samples are also reviewed. These lipids are assumed to be lipids from unidentified or ancient archaea or related organisms. In the second part of this paper, taxonomic and ecological aspects are discussed. Another aspect of archaeal lipid study has to do with its physiological significance, particularly the phase behavior and permeability of archaeal lipid membranes in relation to the thermophily of many archaea. In the last part of this review we discuss this problem. PMID- 16306682 TI - Taka-amylase A in the conidia of Aspergillus oryzae RIB40. AB - A study of Taka-amylase A of conidia from Aspergillus oryzae RIB40 was done. During the research, proteins from conidia and germinated conidia were analyzed using SDS-PAGE, 2-D gel electrophoresis, Western blot analysis, MALDI-TOF Mass spectrometry, and native-PAGE combined with activity staining of TAA. The results showed that TAA exists not only in germinated conidia but also in conidia. Some bands representing degraded products of TAA were detected. Conidia, which formed on starch (SCYA), glucose (DCYA), and glycerol (GCYA) plates, contained mature TAA. Only one active band of TAA was detected after native-PAGE activity staining. In addition, TAA activity was detected in cell extracts of conidia using 0.5 M acetate buffer, pH 5.2, as extraction buffer, but was not detected in whole conidia or cell debris. The results indicate that TAA exists in conidia in active form even when starch, glucose, or glycerol is used as carbon source. TAA might belong to a set of basal proteins inside conidia, which helps in imbibition and germination of conidia. PMID- 16306683 TI - Immunostimulatory effects of a polysaccharide-rich substance with antitumor activity isolated from black currant (Ribes nigrum L.). AB - The fruit juice of black currant was found to contain a polysaccharide-rich substance, which was designated cassis polysaccharide (CAPS), with macrophage stimulating activity. Especially, its interleukin (IL)-1beta-inducing activity was remarkably high, compared with other fruit juice preparations. CAPS was found to consist of rhamnose, mannose, arabinose, galactose, xylose, and glucose in a molar ratio of 11.3:0.9:54.1:29.8:2.0:1.9. CAPS turned out to be partitioned into a soluble component (CAPS-l.m.) and a precipitable component (CAPS-h.m.) with mean MWs of 80,000 and 600,000 respectively in 45% (v/v) ethanol solution. At least in vitro, CAPS-l.m. rather than CAPS-h.m. appeared to play an important role in macrophage activation. Oral administration of black currant juice and CAPS to Ehrlich carcinoma-bearing mice retarded the growth of the solid tumor by 45% and 51% respectively. CAPS administration had a stimulatory effect on the release of IL-2, IL-10, interferon-gamma, and IL-4 from splenocytes in comparison with PBS treatment in tumor-bearing mice. The IL-4 level was, however, still lower than that exhibited by a group of normal mice. CAPS showed a certain cytotoxicity directly against tumor cells. PMID- 16306684 TI - Fluorescent phospholipid analogs as microscopic probes for detection of the mycolic acid-containing layer in Corynebacterium glutamicum: detecting alterations in the mycolic acid-containing layer following ethambutol treatment. AB - Corynebacterium glutamicum belongs to the mycolic acid-containing actinomycetes, which also include Mycobacterium, Nocardia, and Rhodococcus. The cells of this group possess a cell wall with a thick outer layer composed primarily of mycolic acid, which functions as a permeability barrier. To investigate the mechanism of mycolic acid-containing layer (mycolate layer) formation, we have developed a fluorescence microscopic technique detecting the mycolate layer in situ. The staining specificity of fluorescence-labeled phospholipid analogs was determined by simultaneous staining with the hydrophobic fluorescent dye Nile Red and peptidoglycan-staining fluorescence-conjugated vancomycin. We found that fluorescence-labeled phospholipid analogs preferentially stain the mycolate layer. Using this technique, we observed the effect of the anti-mycobacterial drug ethambutol on C. glutamicum mycolate-layer formation. Ethambutol interfered specifically with mycolate-layer formation on the division planes and cell poles, while the side-wall mycolate layer was not severely affected. This indicates that mycolate-layer formation occurs mainly on division planes and cell poles in C. glutamicum, where the peptidoglycan layer is actively synthesized. PMID- 16306685 TI - Interaction between chitosan and oil under stomach and duodenal digestive chemical conditions. AB - Chitosan, the N acetylated derivative of chitin, has an effect on the absorption of dietary lipids, but there is not enough scientific knowledge about the mechanism. To study the interaction between chitosan and oil, the action of this biopolymer has been evaluated through an experimental model of the stomach and duodenum tract, although the enzimatic activity had not been evaluated. We microscopically confirmed that chitosan in a hychloridic acid medium (pH 1.0-2.0) emulsified lipids and the emulsion was a water in oil in water type (w/o/w). When the pH value and speed of agitation were increased to mirror the duodenum medium conditions under which lipids are absorbed, the emulsion capacity was better with an increased number of droplets and the emulsion continued as the w/o/w type. At pH 6.2, chitosan precipitated and lipids were entrapped in the formed flocculus. The binding oil was quantitatively determined, and we also demonstrate that a larger oil quantity induced less retention, while the chitosan characteristics had no influence. These observations allow us to postulate that the interaction between chitosan and oil inhibited duodenal absorption and enhanced lipid excretion. PMID- 16306686 TI - A novel 29-kDa crystal protein from Bacillus thuringiensis induces caspase activation and cell death of jurkat T cells. AB - Bacillus thuringiensis, the most successful and most widely used microbial insecticide, produces crystal proteins. The physiological significance of the crystal proteins is poorly understood except for the potent insecticidal activity. In this paper, we report a novel biological activity of the crystal protein. A 29-kDa crystal protein, p29, produced by B. thuringiensis subsp. coreanensis A1519, was toxic to Jurkat, a cell line from human leukemic T cells. Upon treatment of the Jurkat cells with p29 at a lower concentration, translocation of phosphatidylserine to the external cell surface, release of cytochrome c and Smac/DIABLO from the mitochondria, and activation of caspase-9 were induced. These cellular events were followed by activation of caspase-3, cleavage of poly (ADP-ribose) polymerase (PARP), and chromatin condensation. Peak activation of caspase-9 was prominent and preceded that of caspase-8. Depletion of Bax from the cytosol was observed as the progress of p29-induced cell death. At a higher concentration of p29, the cells showed similar and accelerated morphological change, but neither externalized phosphatidylserine nor caspase-3 activation was observed. These results suggest that p29 at the lower concentration induced cell death of Jurkat accompanied by apotosis-like cellular events, and that mitochondria played a major role in p29-induced cell death. PMID- 16306687 TI - Growth inhibition by tungsten in the sulfur-oxidizing bacterium Acidithiobacillus thiooxidans. AB - Growth of five strains of sulfur-oxidizing bacteria Acidithiobacillus thiooxidans, including strain NB1-3, was inhibited completely by 50 microM of sodium tungstate (Na(2)WO(4)). When the cells of NB1-3 were incubated in 0.1 M beta-alanine-SO(4)(2-) buffer (pH 3.0) with 100 microM Na(2)WO(4) for 1 h, the amount of tungsten bound to the cells was 33 microg/mg protein. Approximately 10 times more tungsten was bound to the cells at pH 3.0 than at pH 7.0. The tungsten binding to NB1-3 cells was inhibited by oxyanions such as sodium molybdenum and ammonium vanadate. The activities of enzymes involved in elemental sulfur oxidation of NB1-3 cells such as sulfur oxidase, sulfur dioxygenase, and sulfite oxidase were strongly inhibited by Na(2)WO(4). These results indicate that tungsten binds to NB1-3 cells and inhibits the sulfur oxidation enzyme system of the cells, and as a result, inhibits cell growth. When portland cement bars supplemented with 0.075% metal nickel and with 0.075% metal nickel and 0.075% calcium tungstate were exposed to the atmosphere of a sewage treatment plant containing 28 ppm of H(2)S for 2 years, the weight loss of the portland cement bar with metal nickel and calcium tungstate was much lower than the cement bar containing 0.075% metal nickel. PMID- 16306688 TI - Detection of hydrogen gas-producing anaerobes in refuse-derived fuel (RDF) pellets. AB - Recently, we reported that refuse-derived fuel (RDF) pellets contain a relatively high number of viable bacterial cells and that these bacteria generate heat and hydrogen gas during fermentation under wet conditions. In this study we analyzed bacterial cell numbers of RDF samples manufactured with different concentrations of calcium hydroxide, which is usually added to waste materials for the prevention of rotting of food wastes and the acceleration of drying of solid wastes, and determined the amount of hydrogen gas produced by them under wet conditions. Furthermore, we analyzed microflora of the RDF samples before and during fermentation by denaturing gradient gel electrophoresis of 16S rDNA followed by sequencing. We found that the RDF samples contained various kinds of clostridia capable of producing hydrogen gas. PMID- 16306689 TI - Proteomic studies of isoforms of the P25 component of Bombyx mori fibroin. AB - It is recognized that P25 is one of three polypeptide components of the fibroin synthesized in the larval silk gland (SG) of silkworm, having two glycosylated isoforms. In the present study, however, eight P25 isoforms were separated by proteomics, including two-dimensional gel electrophoresis of whole SG proteins, and were identified by the peptide mass fingerprinting method. Four of the eight isoforms were identified as Bombyx mandarina P25s, although the SG of Bombyx mori has never been considered to contain the P25 from B. mandarina. It is suggested that this diversity of P25 isoforms depends on phosphorylation modification in addition to glycosylation. PMID- 16306690 TI - Inactivation of enzymes in fresh sake using a continuous flow system for high pressure carbonation. AB - The Inactivation kinetics of alpha-glucosidase, glucoamylase, alpha-amylase, and acid carboxypeptidase in fresh sake using a continuous flow system for high pressure carbonation were investigated. In addition, the effects of ethanol and sugar concentrations on inactivation of the enzymes in high-pressure carbonated sake were investigated. Among the enzymes investigated, alpha-glucosidase was the most stable and alpha-amylase was the most labile on inactivation under carbonation. The decimal reduction times (D values) of alpha-glucosidase, glucoamylase, alpha-amylase (extrapolated from the Z value), and acid carboxypeptidase were 29, 6, 2, and 5 min respectively at 45 degrees C. These values are lower than those subjected to heat treatment. On the carbonation treatment as well as the heat treatment, ethanol accelerated the inactivation of all four enzymes, but glucose depressed the inactivation of these enzymes, except for acid carboxypeptidase. These results suggest that this continuous flow system enabled effective inactivation of enzymes in fresh sake. PMID- 16306691 TI - A single free cysteine residue and disulfide bond contribute to the thermostability of Aspergillus saitoi 1,2-alpha-mannosidase. AB - Aspergillus saitoi 1,2-alpha-mannosidase contains three conserved cysteine residues (Cys334, Cys363, and Cys443). We showed that Cys334 and Cys363 are involved in a disulfide bond, and that Cys443 contains a free thiol group. The cysteines were not essential for the activity analyzed by site-directed mutagenesis and kinetics. The substitution at each cysteine residue greatly destabilized the enzyme. The T(m) values of WT, C443A, C443G, C443S, and C443T were 55.8, 51.9, 50.2, 50.0, and 52.8 degrees C respectively. The specific activity of these mutants was almost equal to that of WT. Introducing Asp, Leu, Met, or Val at position 443 caused partial denaturation, although the enzymes had some activity. C443F, C443I, C443N, and C443Y were not secreted. These results suggest that the hydrophilic and large side chain causes the destabilization. Molecular modelling showed that the Cys443 residue is buried and surrounded by a hydrophobic environment. Cys334 and Cys363 form a disulfide bond, and Cys443 is involved in a hydrophobic interaction to stabilize the enzyme. PMID- 16306692 TI - Mitochondrial ABC transporter Atm1p is required for protection against oxidative stress and vacuolar functions in Schizosaccharomyces pombe. AB - A potential correlation between mitochondrial and vacuolar functions is known to exit in yeast. Fission yeast atm1(+), SPAC15A10.01, encodes a putative half-type ABC transporter with an N-terminal mitochondrial-targeting signal. In an attempt to evaluate the possible involvement of mitochondrion in vacuole function, a functional analysis of atm1(+) was performed by gene disruption. Growth of the atm1 mutant was inhibited in the presence of oxidizing agents, and S. cerevisiae Atm1p was found to complement this growth defect. atm1Delta cells exhibited defects in fluid-phase endocytosis and vacuolar fusion under hypotonic stress. GFP-tagged Atm1p was observed to be localized in the mitochondria. These data strongly suggest that fission yeast Atm1p was not only involved in protection against oxidative stress, but also played a role in vacuolar functions. PMID- 16306693 TI - Up-regulation of the claudin-6 gene in adipogenesis. AB - To investigate the role of claudin-6 in adipogenesis, claudin-6 mRNA was examined in adipose tissues and adipocyte differentiation. Claudin-6 mRNA was found to be differentially expressed in four different adipose tissues, and up-regulated in each fat depot of mice fed a high-fat diet as compared to a normal-fat diet. Levels of claudin-6 transcripts were increased during differentiation of 3T3-L1 cells in vitro. Moreover, small interfering RNA (siRNA)-mediated reduction of claudin-6 mRNA inhibited differentiation of 3T3-L1 cells. These results suggest that claudin-6 is another important regulator in adipogenesis and fat deposition. PMID- 16306694 TI - Analysis of carotenoid composition in petals of calendula (Calendula officinalis L.). AB - Nineteen carotenoids were identified in extracts of petals of orange- and yellow flowered cultivars of calendula (Calendula officinalis L.). Ten carotenoids were unique to orange-flowered cultivars. The UV-vis absorption maxima of these ten carotenoids were at longer wavelengths than that of flavoxanthin, the main carotenoid of calendula petals, and it is clear that these carotenoids are responsible for the orange color of the petals. Six carotenoids had a cis structure at C-5 (C-5'), and it is conceivable that these (5Z)-carotenoids are enzymatically isomerized at C-5 in a pathway that diverges from the main carotenoid biosynthesis pathway. Among them, (5Z,9Z)-lycopene (1), (5Z,9Z,5'Z,9'Z)-lycopene (3), (5'Z)-gamma-carotene (4), and (5'Z,9'Z)-rubixanthin (5) has never before been identified. Additionally, (5Z,9Z,5'Z)-lycopene (2) has been reported only as a synthesized compound. PMID- 16306695 TI - Browning of model orange juice solution: factors affecting the formation of decomposition products. AB - A model solution of orange juice was prepared and stored. Factors affecting browning and formation of such decomposition products as 3-hydroxy-2-pyrone (3OH2P), 5-hydroxymethylfurfural (HMF), furfural, 5-hydroxymaltol, and 2-furoic acid were examined. Ascorbic acid (AsA) was essential for browning, which was stimulated by amino acids and citric acid, and repressed by chelators and radical scavengers (RS). 3OH2P was derived from AsA. Its formation was stimulated by sugars and repressed by citric acid, chelating agents, and RS. HMF was derived from fructose. Furfural was derived from AsA, and its formation was stimulated by sugars and chelating agents and repressed by RS. 5-hydroxymaltol and 2-furoic acid were derived from fructose and AsA respectively. We did not find any decomposition products showing the same formation pattern as the browning, but a furfural solution with added amino acids turned brown like the model orange juice solution. It might be an indicator for the browning of orange juice. PMID- 16306696 TI - A novel thermophilic pectate lyase containing two catalytic modules of Clostridium stercorarium. AB - The Clostridium stercorarium F-9 pel9A gene encodes a pectate lyase Pel9A consisting of 1,240 amino acids with a molecular weight of 135,171. The mature form of Pel9A is a modular enzyme composed of two family-9 catalytic modules of polysaccharide lyases, CM9-1 and CM9-2, in order from the N terminus. Pel9A showed an overall sequence similarity to the hypothetical pectate lyase PelX of Bacillus halodurans (sequence identity 53%), and CM9-2 showed moderate sequence similarities to some pectate lyases of family 9. Sequence identity between CM9-1 and CM9-2 was 21.3%. The full-length Pel9A lacking the N-terminal signal peptide was expressed, purified, and characterized. The enzyme required Ca(2+) ion for its enzyme activity and showed high activity toward polygalacturonic acid but lower activity toward pectin, indicating that Pel9A is a pectate lyase. Immunological analysis using an antiserum raised against the purified enzyme indicated that Pel9A is constitutively synthesized by C. stercorarium F-9. PMID- 16306697 TI - Purification, characterization, and overexpression of psychrophilic and thermolabile malate dehydrogenase of a novel antarctic psychrotolerant, Flavobacterium frigidimaris KUC-1. AB - We purified the psychrophilic and thermolabile malate dehydrogenase to homogeneity from a novel psychrotolerant, Flavobacterium frigidimaris KUC-1, isolated from Antarctic seawater. The enzyme was a homotetramer with a molecular weight of about 123 k and that of the subunit was about 32 k. The enzyme required NAD(P)(+) as a coenzyme and catalyzed the oxidation of L-malate and the reduction of oxalacetate specifically. The reaction proceeded through an ordered bi-bi mechanism. The enzyme was highly susceptible to heat treatment, and the half-life time at 40 degrees C was estimated to be 3.0 min. The k(cat)/K(m) (microM(-1).s( 1)) values for L-malate and NAD(+) at 30 degrees C were 289 and 2,790, respectively. The enzyme showed pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of the coenzyme. The enzyme contained 311 amino acid residues and much lower numbers of proline and arginine residues than other malate dehydrogenases. PMID- 16306698 TI - Functional analysis of the YvrGHb two-component system of Bacillus subtilis: identification of the regulated genes by DNA microarray and northern blot analyses. AB - YvrGHb is a novel two-component system in Bacillus subtilis. The yvrG and yvrHb genes are considered to encode sensor kinase and response regulator respectively. We found that the YvrGHb system positively regulates the 7 transcriptional units (wprA, wapA-yxxG, dltABCDE, sunA, sunT-bdbA-yolJ-bdbB, yvrI-yvrHa, and sigX rsiX), and negatively regulates the lytABC operon. wprA, wapA, lytB, and lytC encode the main cell surface proteins of B. subtilis. Furthermore, SigX [extracytoplasmic function sigma factor (ECF sigma factor)] relating to the cell surface homeostatic functions, was regulated by this system. The yvrGHb null mutant showed the unusual autolysis and higher susceptibility to the four kinds of antibiotics (aztreonam, cefepime, bacitracin, and fosfomycin). These results indicate that the YvrGHb system is important to maintain the overall state of the cell surface through transcriptional regulation. PMID- 16306699 TI - Mode of action of beta-glucuronidase from Aspergillus niger on the sugar chains of arabinogalactan-protein. AB - A beta-glucuronidase purified from a commercial pectolytic enzyme preparation of Aspergillus niger hydrolyzed about half of the 4-O-methyl-glucuronic acid (4-Me GlcA) residues located at the nonreducing terminals of (1-->6)-linked beta galactosyl side chains of the carbohydrate portion of a radish arabinogalactan protein (AGP) modified by treatment with fungal alpha-L-arabinosidase. Digestion of the alpha-L-arabinosidase-treated AGP with exo-beta-(1-->3)-galactanase released, by exo-fission of beta-(1-->3)-galactosidic bonds in the backbone chains of the AGP, neutral beta-(1-->6)-galactooligosaccharides with various chain lengths and their acidic derivatives substituted at their nonreducing terminals with 4-Me-beta-GlcA groups. In contrast, successive digestion of the alpha-L-arabinosidase-treated AGP with beta-glucuronidase followed by exo-beta-(1 ->3)-galactanase liberated much higher amounts of beta-(1-->6)-galactooligomers together with a small portion of short acidic oligomers, mainly 4-Me-beta-GlcA-(1 ->6)-Gal and 4-Me-beta-GlcA-(1-->6)-beta-Gal-(1-->6)-Gal. These results indicate that beta-glucuronidase acts upon 4-Me-beta-GlcA residues in long (1-->6)-linked beta-galactosyl side chains of the AGP, whereas short acidic side chains survive the attack of the enzyme. PMID- 16306700 TI - Cloning and characterization of saponin hydrolases from Aspergillus oryzae and Eupenicillium brefeldianum. AB - We purified saponin hydrolases from Aspergillus oryzae PF1224 and Eupenicillium brefeldianum PF1226. It was confirmed that the enzymes from A. oryzae PF1224 (Sda1) and E. brefeldianum PF1226 (Sde1) are glycoproteins with molecular masses of 82 and 90 kDa respectively. The deduced amino acid sequences of each enzyme from the cloned genes (sda1 or sde1) showed approximately 50% homology with that of the saponin hydrolase Sdn1 from Neocosmospora vasinfecta var. vasinfecta PF1225 (DDBJ accession no. AB110615). When sda1 and sde1 were expressed in the host Trichoderma viride under the control of the cellobiohydrolase I gene promoter, recombinant proteins were secreted with molecular masses of 77 and 67 kDa respectively. These recombinant enzymes hydrolyzed soyasaponin I to soyasapogenol B and triose, and its substrate specificities for glycosides were similar to that of Sdn1, but the specific activities of these enzymes were lower than that of Sdn1. PMID- 16306701 TI - Isolation and structure determination of algicidal compounds from Ulva fasciata. AB - Thirty-seven species of seaweeds including 10 Chlorophyta, 13 Phaeophyta, and 14 Rhodophyta collected from the coast of Nagasaki Prefecture, Japan, were screened for algicidal activity against the red-tide phytoplankton Heterosigma akashiwo. The green alga Ulva fasciata (Ulvaceae, Chlorophyta) showed the strongest algicidal activity among the seaweeds tested. Bioassay-guided fractionation of the methanol extract of U. fasciata led to isolation of three algicidal compounds whose structures were determined to be hexadeca-4,7,10,13-tetraenoic acid (HDTA), octadeca-6,9,12,15-tetraenoic acid (ODTA), and alpha-linolenic acid on the basis of spectroscopic information. These polyunsaturated fatty acids (PUFAs) showed potent algicidal activity against H. akashiwo (LC(50) 1.35 microg/ml, 0.83 microg/ml, and 1.13 microg/ml for HDTA, ODTA, and alpha-linolenic acid, respectively), and the result demonstrated the potential of these PUFAs for practical harmful algal bloom control. PMID- 16306702 TI - Botanical and geographical origin identification of industrial ethanol by stable isotope analyses of C, H, and O. AB - The isotope ratios of carbon, hydrogen, and oxygen of rectified alcohols were determined to distinguish their botanical and geographical origins by continuous flow-isotope ratio mass spectrometry (CF-IRMS). The (13)C/(12)C and (18)O/(16)O ratios of 27 fermented alcohols with known origins showed clusters derived from each botanical origin, viz. corn, sugarcane, wheat, and tapioca. C3 and C4 plants were easily distinguishable by the (13)C/(12)C ratio. Sugarcane and corn are both C4 plants, and they showed small differences in isotope ratios. The combination plots of the D/H and (18)O/(16)O ratios enabled us to designate the geographical origins of alcohol derived from the same kind of crop, such as Chinese or American corn. The chemically synthetic and fermented alcohols were clearly distinguished by D/H and (18)O/(16)O ratios. Isotope ratios were useful for origin identification of alcohol. We plan to construct a database of alcohol isotope ratios to determine the origins of raw materials in alcohol. PMID- 16306703 TI - Enzymic activity of the K5-type yeast killer toxin and its characterization. AB - K5-type yeast killer toxin secreted by P. anomala NCYC 434 cells has a broad killing spectrum. Competitive inhibiton of killer activity showed that glucans, mainly the beta-1,3 glucan, represent the primary toxin binding site within the cell wall of sensitive cells. Its hydrolytic activity on laminarin in an exo-like fashion revealed that the toxin exerts its killing effect by exo-beta-1,3 glucanase activity. Its specific activity on laminarin was 120 U/mg, and the Michaelis constants K(m) and V(max) for laminarin hydrolysis were 0.25 mg/ml and 370 micromol/min/mg. The toxin exerted its cytocidal effect after 2 h contact with the target cells. Production of the toxin by the cells was induced only when they were grown in culture media rich in beta-glucan sources, and the addition of glucose increased the specific production rate. The enzymic activity of the toxin was fully inhibited by Hg(+2), but increased with some other metal ions, most of all by Pb(+2). PMID- 16306704 TI - Phorbol myristate acetate induces neutrophil death through activation of p38 mitogen-activated protein kinase that requires endogenous reactive oxygen species other than HOCl. AB - Stimulation of normal mouse neutrophils with phorbol 12-myristate 13-acetate resulted in an acceleration of chromatin condensation and phosphatidylserine externalization that was not associated with caspase-3 activation. Caspase independent death was completely inhibited by GF109203X and SB202190, specific inhibitors for protein kinase C and p38 mitogen-activated protein kinase respectively. Activation of p38 mitogen-activated protein kinase was completely suppressed by GF109203X, indicating that this enzyme is regulated by protein kinase C. On the other hand, cell death was abolished in NADPH oxidase-deficient neutrophils lacking superoxide production. Of note, p38 mitogen-activated protein kinase was activated by phorbol 12-myristate 13-acetate in normal and myeloperoxidase-deficient neutrophils lacking production of HOCl, whereas no activation was observed in NADPH oxidase-deficient neutrophils. These results strongly suggest that activation of p38 mitogen-activated protein kinase is regulated by endogenously generated superoxide or its metabolites other than HOCl, a critical regulator of inducer-stimulated death of neutrophils. PMID- 16306705 TI - Fredericamycin A affects mitochondrial inheritance and morphology in Saccharomyces cerevisiae. AB - Fredericamycin A (FMA) is an antibiotic product of Streptomyces griseus that exhibits modest antitumor activity in vivo and in vitro, but, its functions in vivo are poorly understood. We identified this compound as an inducer of G1 arrest in the yeast, Saccharomyces cerevisiae. FMA exhibits an IC50 of 24 nM towards the growth of a disruptant of multi-drug resistance genes, W303-MLC30, and its cytotoxicity is a function of the time of exposure as well as drug dose. Addition of 0.8 microM of FMA caused aggregation of mitochondria within 10 min of incubation and the drug induced petites at high frequency after 4 h of incubation. Rho(-) cells were about 20 times more resistant to FMA than isogenic rho(+) cells. Overexpression of topoisomerase I, a previously suggested target of the drug, did not alleviate the sensitivity of the cells to FMA or the aggregation of mitochondria. Our results suggest that mitochondria are the primary target site of FMA. PMID- 16306706 TI - Effect of caffeine on the body fat and lipid metabolism of rats fed on a high-fat diet. AB - The intake of caffeine (CF) at 0.025, 0.05 or 0.1% for 21 days progressively reduced the body fat mass and body fat percentage in Sprague-Dawley (SD) rats fed on a high-fat diet with increasing administration level. Moreover, CF increased the serum concentrations of catecholamines and free fatty acids in SD rats orally administered with CF (5 mg/kg). These results suggest that the intake of CF reduced body fat by lipolysis via catecholamines. CF has potential as a functional food ingredient with an anti-obesity action. PMID- 16306707 TI - Characteristics of wines made by Saccharomyces mutants which produce a polygalacturonase under wine-making conditions. AB - Wines by yeast mutants producing polygalacturonase in high glucose concentration, from Saccharomyces wine-making strains, had higher filterability and more concentrated anthocyanin contents than that of their parent strains. These results suggest that the clarification process was improved at a lower cost by the low viscosity and that high-quality wines result from the increase in the anthocyanin contents. PMID- 16306708 TI - Inhibitory effect of Aucubin isolated from Eucommia ulmoides against UVB-induced matrix metalloproteinase-1 production in human skin fibroblasts. AB - Of 30 herbal plants tested, the methanol extracts of Eucommia ulmoides (52%), Evodia officinalis (45%), and Pleuropterus multiflorus (41%) each showed a potent inhibitory effect on the matrix metalloproteinase-1 (MMP-1) production in ultraviolet B (UVB)-irradiated human fibroblasts. Aucubin was isolated as the MMP 1 inhibitor from E. ulmoides, and significantly suppressed the production of MMP 1 by nearly 57% compared to the control. It also reduced MMP-1 mRNA expression. These results suggest that aucubin is a photoprotective phytochemical, and could be used as a potential agent in preventing photoaging. PMID- 16306709 TI - Isolation of mutagenic beta-carboline derivatives after nitrite treatment of maillard reaction mixtures and analysis of these compounds from foodstuffs and human urine. AB - Mixtures of carbohydrate decomposition products and L-tryptophan were incubated at pH 7.0 and 37 degrees C for 4 weeks. These mixtures exhibited mutagenic activity toward S. typhimurium TA 100 without metabolic activation after a nitrite treatment at pH 4.0. Four beta-carboline derivatives were isolated as premutagens from mixtures of methylglyoxal and furfural. These premutagens were also found to be contained in daily foodstuffs and human urine samples. PMID- 16306710 TI - A role of androgen receptor protein in cell growth of an androgen-independent prostate cancer cell line. AB - Prostate cancer, which develops due to androgen and is initially responsive to androgen deprivation therapy, often comes to acquire androgen deprivation therapy resistance in short order. We investigated the role of androgen receptor (AR) protein in an androgen-independent prostate cancer cell line using AR ligands and AR siRNA. Although the androgen-independent cell line scarcely responded to AR ligands, their growth was attenuated by ablation of AR protein by siRNA. PMID- 16306711 TI - Single microspore reverse transcription-polymerase chain reaction: dynamic change of gene expression during microsporogenesis. AB - A sensitive and reliable single microspore reverse transcription-polymerase chain reaction (RT-PCR) assay was developed that requires little sample material and only a short sampling time and ensures template preservation. We propose that the assay provides a useful tool to study not only cell-cycle related events in developing pollen but also the molecular mechanisms underlying male sterility and mutants impaired in pollen development. PMID- 16306712 TI - Homogeneous enzymatic assay for L-cysteine with betaC-S lyase. AB - We have developed a new enzymatic assay for determining L-cysteine concentration. The method involves the use of betaC-S lyase from Streptococcus anginosus, which catalyzes the alpha,beta-elimination of L-cysteine to hydrogen sulfide, pyruvate, and ammonia. The production of pyruvate is measured by D-lactate dehydrogenase and NADH. The decrease in NADH was proportional to the L-cysteine concentration up to 1.0 mM. When serum samples were used, within-day and day-to-day coefficient variations were below 4%. This method is simple, and can easily and reliably be used for accurate determination of L-cysteine concentration in serum or other samples. PMID- 16306713 TI - Analysis of recQ gene transcript in fruiting bodies of basidiomycetous mushroom Lentinula edodes. AB - Quantitative reverse transcription PCR analysis and total RNA staining demonstrated that Lentinula edodes recQ gene (Le.recQ) transcript is present in all the parts of the fruiting body, but in hymenophore at the highest density. Results of in situ RNA-RNA hybridization showed that the Le.recQ transcript level within the hymenophore is higher in the hymenium, subhymenium, and the outer region of the trama. Trama cells themselves contain a lower level of the transcript. PMID- 16306714 TI - Synthesis and biological activity of fatty acid derivatives of quinine. AB - Derivatives of quinine with fatty acids including polyunsaturated fatty acids were prepared. They showed moderate antimalarial activity as compared with quinine itself using Plasmodium falciparum. The activities were not dependent on whether the fatty acyl group was saturated or unsaturated. On the other hand, the derivatives showed significantly higher cytotoxicity against a mammary tumor cell line FM3A than quinine itself. Calculating from these data, an acetyl derivative of quinine with the shortest acyl group was found to give the highest selectivity. PMID- 16306715 TI - Purification and characterization of aldoxime dehydratase of the head blight fungus, Fusarium graminearum. AB - Fungal aldoxime dehydratase (Oxd) of Fusarium graminearum MAFF305135 was purified and characterized for the first time from its overexpressing Escherichia coli transformant. The enzyme showed about 20% identity with known Oxds, and had similar enzymatic properties with nitrilase-linked Oxd from the Bacillus strain. It belongs to a group of phenylacetaldoxime dehydratases (EC 4.99.1.7), based on its substrate specificity and kinetic analysis. PMID- 16306716 TI - Apolipoprotein complexity in Japanese eel Anguilla japonica: truncated apolipoprotein A-I and apolipoprotein A-I-like protein in plasma lipoproteins. AB - A truncated apolipoprotein (apo) A-I with a molecular weight (M(r)) of 26 kDa was first isolated from the plasma high density lipoproteins of an atypical Japanese eel (Anguilla japonica). Interestingly, this eel contained a very small amount of intact apoA-I (M(r)28 kDa) in the plasma, although serine protease inhibitors were present throughout the plasma preparation. The N-terminal sequence of 20 amino acids in truncated apoA-I was completely identical with that of intact apoA I. Another apolipoprotein with M(r)28 kDa, whose N-terminal amino acid sequence differed from apoA-I, was also found in high density lipoprotein and low density lipoprotein. The apolipoprotein profiles of Japanese eel plasma appear to be complicated. PMID- 16306720 TI - Epidural analgesia and breast-feeding. PMID- 16306721 TI - Spinal anesthesia with an indwelling catheter reduces the stress response in pediatric open heart surgery. AB - BACKGROUND: Extreme stress and inflammatory responses to open heart surgery are associated with increased morbidity and mortality. Based on both animal and adult human data, it was hypothesized that spinal anesthesia would be more effective at attenuating these responses than conventional high dose intravenous opioid techniques in infants and young children undergoing open heart surgery. METHODS: A prospective randomized controlled clinical trial was performed in 60 children aged up to 24 months undergoing open heart surgery. Patients were randomly assigned to receive either high-dose intravenous opioid or high-dose intravenous opioid plus spinal anesthesia. Spinal anesthesia was administered via an indwelling intrathecal catheter. RESULTS: Spinal anesthesia significantly reduced the stress responses as measured by plasma norepinephrine and epinephrine concentrations (both P < 0.05). Spinal anesthesia reduced plasma lactate concentrations (P < 0.05), but increased fluid requirements during the first postoperative day (P < 0.05). There were no differences in other cardiovascular parameters. CONCLUSIONS: Continuous spinal anesthesia reduces stress responses in infants and young children undergoing cardiac surgery with cardiopulmonary bypass more effectively than high-dose intravenous opioids alone. PMID- 16306722 TI - Validity of unplanned admission to an intensive care unit as a measure of patient safety in surgical patients. AB - BACKGROUND: An unplanned admission to the intensive care unit within 24 h of a procedure (UIA) is a recommended clinical indicator in surgical patients. Often regarded as a surrogate marker of adverse events, it has potential as a direct measure of patient safety. Its true validity for such use is currently unknown. METHODS: The authors validated UIA as an indicator of safety in surgical patients in a prospective cohort study of 44,130 patients admitted to their hospital. They assessed the association of UIA with intraoperative incidents and near misses, increased hospital length of stay, and 30-day mortality as three constructs of patient safety. RESULTS: The authors identified 201 patients with a UIA; 104 (52.2%) had at least one incident or near miss. After adjusting for confounders, these incidents were significantly associated with UIA in all categories of surgical procedures analyzed; odds ratios were 12.21 (95% confidence interval [CI], 6.33-23.58), 4.06 (95% CI, 2.74-6.03), and 2.13 (95% CI, 1.02-4.42), respectively. The 30-day mortality for patients with UIA was 10.9%, compared with 1.1% in non-UIA patients. After risk adjustment, UIA was associated with excess mortality in several types of surgical procedures (odds ratio, 3.89; 95% CI, 2.14 7.04). The median length of stay was increased if UIA occurred: 16 days (interquartile range, 10-31) versus 2 days (interquartile range, 0.5-9) (P < 0.001). For patients with a UIA, the likelihood of discharge from hospital was significantly decreased in most surgical categories analyzed, with adjusted hazard ratios of 0.41 (95% CI, 0.23-0.77) to 0.58 (95% CI, 0.37-0.93). CONCLUSIONS: These findings provide strong support for the construct validity of UIA as a measure of patient safety. PMID- 16306723 TI - Behavioral interactions in the perioperative environment: a new conceptual framework and the development of the perioperative child-adult medical procedure interaction scale. AB - BACKGROUND: The authors suggest that research in the area of parental presence during induction of anesthesia should shift to emphasize what parents actually do during induction, rather than focusing simply on their presence. As a first step, the authors aimed to develop a behavioral coding system that would measure child and adult interactions in the perioperative environment. METHODS: The authors enrolled 45 parents and children (aged 2-12 yr) undergoing elective surgery and general anesthesia. A multidisciplinary team examined videotapes and transcriptions of interactions between children, parents, and medical personnel in the holding room and operating room. The team used an existing scale, the Child-Adult Medical Procedure Interaction Scale, as the prototype for the development of a new perioperative behavioral coding system. The research team conducted extensive revisions to the original scale and added multiple codes to the original scale, including nonverbal codes. Interrater reliability was assessed using weighted kappa statistics. Construct validity was also examined. RESULTS: The final Perioperative Child-Adult Medical Procedure Interaction Scale contains 40 codes in four domains. Analyses showed excellent reliability overall for verbal and nonverbal codes. Kappa values averaged 0.87 for verbal codes characterizing adult vocalizations, 0.92 for verbal codes characterizing child vocalizations, and 0.88 for nonverbal codes. Construct validity was demonstrated by finding the hypothesized associations between certain scale codes and children's anxiety (P = 0.0001). CONCLUSION: Showing excellent reliability, the Perioperative Child-Adult Medical Procedure Interaction Scale is an appropriate tool for assessing child-adult behavioral interaction during the perioperative period. When sequential analyses are conducted and target behaviors are identified, empirically based parent preparation programs can be developed. PMID- 16306724 TI - Optimal puncture site of the right internal jugular vein after laryngeal mask airway placement. AB - BACKGROUND: Puncturing of the internal jugular vein (IJV) after placement of a laryngeal mask airway (LMA) is difficult. Overlapping of the right internal jugular vein (IJV) and common carotid artery (CCA) after placement of an LMA Classic (Laryngeal Mask Company, Henley-on-Thames, United Kingdom) was investigated using an ultrasound scanner. A study was conducted to determine the optimal puncture site in the case of puncturing the right IJV after LMA-Classic placement. METHODS: The subjects in this study consisted of 114 patients (60 men and 54 women) scheduled to undergo LMA-Classic placement (size 4 for men and size 3 for women). Overlapping between the IJV and CCA was investigated at three points (high, middle, and low points) on the right side of the neck after LMA Classic placement. A test puncture was also made at the right low (supraclavicular) point after LMA-Classic placement. RESULTS: (1) The degree of overlapping of the right IJV and CCA after LMA-Classic placement was clearly greater than before LMA placement. (2) With the exception of some measurements, there were many cases in which the right IJV and CCA were completely overlapping after LMA-Classic placement. Furthermore, the CCA was not observed in the vicinity of the right IJV at the right low point. (3) There were no complications (including pneumothorax and accidental arterial puncture) during supraclavicular IJV puncture after LMA-Classic placement. CONCLUSION: After placement of the LMA Classic, overlapping of the IJV and CCA increased at the high and middle puncture points of the IJV; however, at the lower puncture point, the position of the vessels remained unaffected. Therefore, during IJV puncture at high or middle points, ultrasound guidance is advisable to avoid CCA puncture. PMID- 16306725 TI - Respiratory reflex responses of the larynx differ between sevoflurane and propofol in pediatric patients. AB - BACKGROUND: The effects of anesthetics on airway protective reflexes have not been extensively characterized in children. The aim of this study was to compare the laryngeal reflex responses in children anesthetized with either sevoflurane or propofol under two levels of hypnosis using the Bispectral Index score (BIS). The authors hypothesized that the incidence of apnea with laryngospasm evoked by laryngeal stimulation would not differ between sevoflurane and propofol when used in equipotent doses and that laryngeal responsiveness would be diminished with increased levels of hypnosis. METHODS: Seventy children, aged 2-6 yr, scheduled to undergo elective surgery were randomly allocated to undergo propofol or sevoflurane anesthesia while breathing spontaneously through a laryngeal mask airway. Anesthesia was titrated to achieve the assigned level of hypnosis (BIS 40 +/- 5 or BIS 60 +/- 5) in random order. Laryngeal and respiratory responses were elicited by spraying distilled water on the laryngeal mucosa, and a blinded reviewer assessed evoked responses. RESULTS: Apnea with laryngospasm occurred more often during anesthesia with sevoflurane compared with propofol independent of the level of hypnosis: episodes lasting longer than 5 s, 34% versus 19% at BIS 40 and 34% versus 16% at BIS 60; episodes lasting longer than 10 s, 26% versus 10% at BIS 40 and 26% versus 6% at BIS 60 (group differences P < 0.04 and P < 0.01, respectively). In contrast, cough and expiration reflex occurred significantly more frequently in children anesthetized with propofol. CONCLUSION: Laryngeal and respiratory reflex responses in children aged 2-6 yr were different between sevoflurane and propofol independent of the levels of hypnosis examined in this study. PMID- 16306726 TI - Intercellular adhesion molecule-1 inhibition attenuates neurologic and hepatic damage after resuscitation in mice. AB - BACKGROUND: Cardiac arrest and cardiopulmonary resuscitation may result in multiorgan damage after global hypoxia due to neutrophil recruitment. Patients display all signs of a systemic inflammatory response syndrome. Reducing neutrophil recruitment may thus preserve organ function. METHODS: Mice were subjected to cardiac arrest and resuscitation. CD18/CD11b expression on circulating neutrophils was assessed by flow cytometry. Intercellular adhesion molecule-1 expression was analyzed by Western blot and immunofluorescence. Neutrophil recruitment was quantified by immunohistochemistry. Neurologic function was assessed by a balance test. For liver and kidney function, plasma alanine aminotransferase activity and creatinine concentrations were determined. To reduce neutrophil recruitment, mice received 100 microg anti-intercellular adhesion molecule-1 antibody intraperitoneally. RESULTS: Resuscitation led to severe hypoxia, acidosis, and hypercarbia. Adhesion molecule expression and neutrophil recruitment were increased in the liver, kidney, and brain. Neurologic performance was impaired 24 h after cardiac arrest. Creatinine and alanine aminotransferase concentrations were significantly increased. Immunoneutralization of intercellular adhesion molecule-1 attenuated neutrophil influx in the liver along with alanine aminotransferase activity, whereas creatinine concentrations and neutrophil influx in the kidney remained unchanged. Neurologic function was improved in the treatment group. CONCLUSIONS: Global hypoxia induces activation of the endothelium in the brain, liver, and kidney. The resulting damage to the brain and liver are due to infiltration of neutrophils, whereas kidney damage is not, because reduction of neutrophil recruitment after cardiopulmonary resuscitation improves recovery of neurologic and hepatic but not renal function. Inhibition of intercellular adhesion molecule 1 after global hypoxia may be beneficial in patients experiencing cardiac arrest and resuscitation. PMID- 16306727 TI - Halothane inhibition of recombinant cardiac L-type Ca2+ channels expressed in HEK 293 cells. AB - BACKGROUND: Volatile anesthetics depress cardiac contractility, which involves inhibition of cardiac L-type calcium channels. To explore the role of voltage dependent inactivation, the authors analyzed halothane effects on recombinant cardiac L-type calcium channels (alpha1Cbeta2a and alpha1Cbeta2aalpha2/delta1), which differ by the alpha2/delta1 subunit and consequently voltage-dependent inactivation. METHODS: HEK-293 cells were transiently cotransfected with complementary DNAs encoding alpha1C tagged with green fluorescent protein and beta2a, with and without alpha2/delta1. Halothane effects on macroscopic barium currents were recorded using patch clamp methodology from cells expressing alpha1Cbeta2a and alpha1Cbeta2aalpha2/delta1 as identified by fluorescence microscopy. RESULTS: Halothane inhibited peak current (I(peak)) and enhanced apparent inactivation (reported by end pulse current amplitude of 300-ms depolarizations [I300]) in a concentration-dependent manner in both channel types. alpha2/delta1 coexpression shifted relations leftward as reported by the 50% inhibitory concentration of I(peak) and I300/I(peak)for alpha1Cbeta2a (1.8 and 14.5 mm, respectively) and alpha1Cbeta2aalpha2/delta1 (0.74 and 1.36 mm, respectively). Halothane reduced transmembrane charge transfer primarily through I(peak) depression and not by enhancement of macroscopic inactivation for both channels. CONCLUSIONS: The results indicate that phenotypic features arising from alpha2/delta1 coexpression play a key role in halothane inhibition of cardiac L type calcium channels. These features included marked effects on I(peak) inhibition, which is the principal determinant of charge transfer reductions. I(peak) depression arises primarily from transitions to nonactivatable states at resting membrane potentials. The findings point to the importance of halothane interactions with states present at resting membrane potential and discount the role of inactivation apparent in current time courses in determining transmembrane charge transfer. PMID- 16306728 TI - Memory enhancing effect of low-dose sevoflurane does not occur in basolateral amygdala-lesioned rats. AB - BACKGROUND: Certain anesthetics might enhance aversive memory at doses around 0.1 minimum alveolar concentration. This issue was investigated in a rat model of learning and memory. In addition, evidence for basolateral amygdala (BLA) involvement in mediating memory enhancement was sought. METHODS: First, the memory-enhancing potential of various anesthetics was determined. Rats underwent single-trial inhibitory avoidance training (0.3 mA shock/1 s) during exposure to air, 0.11% sevoflurane, 0.10% halothane, 0.77% desflurane, or 0.12% isoflurane. Memory was assessed at 24 h. Second, the BLA contribution to sevoflurane memory enhancement was determined. Rats received bilateral excitotoxic N-methyl-D aspartate (12.5 mg in 0.2 microl per BLA) lesions of the BLA 1 week before training. Memory of lesioned and control rats was compared 24 h after training in air or sevoflurane. RESULTS: Sevoflurane exposure during training significantly enhanced 24-h retention performance for both nonoperated and sham-operated rats (P < 0.005 for both vs. their respective controls). Halothane, but not desflurane or isoflurane, also enhanced retention performance (P < 0.05). However, halothane induced hyperalgesia during learning clouds interpreting enhanced retention performance solely as a memory consolidation effect. BLA lesions significantly reduced and equalized retention performance for both sevoflurane- and air-exposed animals. Lesions blocked memory enhancement without also causing a generalized inability to learn, because additional training revealed essentially normal task acquisition and 24-h memory. CONCLUSIONS: Sevoflurane enhances aversive memory formation in the rat. The BLA likely contributes to this effect. The risk of aversive memory formation may be enhanced during exposure to low-dose sevoflurane. PMID- 16306729 TI - Effects of nitrous oxide on the rat heart in vivo: another inhalational anesthetic that preconditions the heart? AB - BACKGROUND: For nitrous oxide, a preconditioning effect on the heart has yet not been investigated. This is important because nitrous oxide is commonly used in combination with volatile anesthetics, which are known to precondition the heart. The authors aimed to clarify (1) whether nitrous oxide preconditions the heart, (2) how it affects protein kinase C (PKC) and tyrosine kinases (such as Src) as central mediators of preconditioning, and (3) whether isoflurane-induced preconditioning is influenced by nitrous oxide. METHODS: For infarct size measurements, anesthetized rats were subjected to 25 min of coronary artery occlusion followed by 120 min of reperfusion. Rats received nitrous oxide (60%), isoflurane (1.4%) or isoflurane-nitrous oxide (1.4%/60%) during three 5-min periods before index ischemia (each group, n = 7). Control animals remained untreated for 45 min. Additional hearts (control, 60% nitrous oxide alone%, and isoflurane-nitrous oxide [0.6%/60%, in equianesthetic doses]) were excised for Western blot of PKC-epsilon and Src kinase (each group, n = 4). RESULTS: Nitrous oxide had no effect on infarct size (59.1 +/- 15.2% of the area at risk vs. 51.1 +/- 10.9% in controls). Isoflurane (1.4%) and isoflurane-nitrous oxide (1.4%/60%) reduced infarct size to 30.9 +/- 10.6 and 28.7 +/- 11.8% (both P < 0.01). Nitrous oxide (60%) had no effect on phosphorylation (2.3 +/- 1.8 vs. 2.5 +/- 1.7 in controls, average light intensity, arbitrary units) and translocation (7.0 +/- 4.3 vs. 7.4 +/- 5.2 in controls) of PKC-epsilon. Src kinase phosphorylation was not influenced by nitrous oxide (4.6 +/- 3.9 vs. 5.0 +/- 3.8; 3.2 +/- 2.2 vs. 3.5 +/- 3.0). Isoflurane-nitrous oxide (0.6%/60%, in equianesthetic doses) induced PKC-epsilon phosphorylation (5.4 +/- 1.9 vs. 2.8 +/- 1.5; P < 0.001) and translocation to membrane regions (13.8 +/- 13.0 vs. 6.7 +/- 2.0 in controls; P < 0.05). CONCLUSIONS: Nitrous oxide is the first inhalational anesthetic without preconditioning effect on the heart. However, isoflurane-induced preconditioning and PKC-epsilon activation are not influenced by nitrous oxide. PMID- 16306730 TI - Influence of sevoflurane on the metabolism and renal effects of compound A in rats. AB - BACKGROUND: The sevoflurane degradation product compound A is nephrotoxic in rats. In contrast, patient exposure to compound A during sevoflurane anesthesia has no clinically significant renal effects. The mechanism for this difference is incompletely understood. One possibility is that the metabolism and toxicity of compound A in humans is prevented by sevoflurane. However, the effect of sevoflurane on compound A metabolism and nephrotoxicity is unknown. Thus, the purpose of this investigation was to determine the effect of sevoflurane on the metabolism and renal toxicity of compound A in rats. METHODS: Male rats received 0.25 mmol/kg intraperitoneal compound A, alone and during sevoflurane anesthesia (3%, 1.3 minimum alveolar concentration, for 3 h). Compound A metabolites in urine were quantified, and renal function was evaluated by serum creatinine and urea nitrogen, urine volume, osmolality, protein excretion, and renal tubular histology. RESULTS: Sevoflurane coadministration with compound A inhibited compound A defluorination while increasing relative metabolism through pathways of sulfoxidation and beta-lyase-catalyzed metabolism, which mediate toxicity. Sevoflurane coadministration with compound A increased some (serum creatinine and urea nitrogen, and necrosis) but not other (urine volume, osmolality, and protein excretion) indices of renal toxicity. CONCLUSIONS: Sevoflurane does not suppress compound A nephrotoxicity in rats in vivo. These results do not suggest that lack of nephrotoxicity in surgical patients exposed to compound A during sevoflurane anesthesia results from an inhibitory effect of sevoflurane on compound A metabolism and toxicity. Rather, these results are consistent with differences between rats and humans in compound A exposure and inherent susceptibility to compound A nephrotoxicity. PMID- 16306731 TI - Influence of hemorrhagic shock and subsequent fluid resuscitation on the electroencephalographic effect of isoflurane in a swine model. AB - BACKGROUND: The authors have previously reported that hemorrhage does not alter the electroencephalographic effect of isoflurane under conditions of compensated hemorrhagic shock. Here, they have investigated the influence of decompensated hemorrhagic shock and subsequent fluid resuscitation on the electroencephalographic effect of isoflurane. METHODS: Twelve swine were anesthetized through inhalation of 2% isoflurane. The inhalational concentration was then decreased to 0.5% and maintained for 25 min, before being returned to 2% and maintained for 25 min (control period). Hemorrhagic shock was then induced by removing 28 ml/kg blood over 30 min. After a 30-min stabilization period, the inhalational concentration was varied as in the control period. Finally, fluid infusion was performed over 30 min using a volume of hydroxyethyl starch equivalent to the blood withdrawn. After a 30-min stabilization period, the inhalational concentration was again varied as in the control period. End-tidal isoflurane concentrations and spectral edge frequency were recorded throughout the study. The pharmacodynamics were characterized using a sigmoidal inhibitory maximal effect model for spectral edge frequency versus effect site concentration. RESULTS: Decompensated hemorrhagic shock slightly but significantly shifted the concentration-effect relation to the left, demonstrating a 1.12-fold decrease in the effect site concentration required to achieve 50% of the maximal effect in the spectral edge frequency. Fluid resuscitation reversed the onset of isoflurane, which was delayed by hemorrhage, but did not reverse the increase in end-organ sensitivity. CONCLUSIONS: Although decompensated hemorrhagic shock altered the electroencephalographic effect of isoflurane regardless of fluid resuscitation, the change seemed to be minimal, in contrast to several intravenous anesthetics. PMID- 16306732 TI - Rapacuronium augments acetylcholine-induced bronchoconstriction via positive allosteric interactions at the M3 muscarinic receptor. AB - BACKGROUND: Neuromuscular blocking agents' detrimental airway effects may occur as a result of interactions with muscarinic receptors, allergic reactions, or histamine release. Rapacuronium, a nondepolarizing muscle relaxant, was withdrawn from clinical use because of its association with fatal bronchospasm. Despite its withdrawal from clinical use, it is imperative that the mechanism by which bronchospasm occurred is understood so that new muscle relaxants introduced to clinical practice do not share these same detrimental airway effects. METHODS: Airway smooth muscle force was measured in guinea pig tracheal rings in organ baths exposed to muscle relaxants with or without subthreshold concentrations of acetylcholine. Antagonism of muscarinic, histamine, neurokinin, leukotriene receptors, or blockade of L-type calcium channels or depletion of nonadrenergic, noncholinergic neurotransmitters was performed. Muscle relaxants' potentiation of acetylcholine-stimulated inositol phosphate synthesis and allosteric interactions on the kinetics of atropine-induced [3H]N-methylscopolamine dissociation were measured in cells expressing recombinant human M3 muscarinic receptors. RESULTS: Rapacuronium, within clinically achieved concentrations, contracted tracheal rings in the presence but not in the absence of subthreshold concentrations of acetylcholine. This effect was prevented or reversed only by atropine. The allosteric action of rapacuronium was demonstrated by the slowing of atropine induced dissociation of [3H]N-methylscopolamine, and positive cooperativity was demonstrated by potentiation of acetylcholine-induced inositol phosphate synthesis. CONCLUSION: Many muscle relaxants have allosteric properties at muscarinic receptors; however, positive cooperativity at the M3 muscarinic receptor within clinically relevant concentrations is unique to rapacuronium. These findings establish novel parameters that should be considered in the evaluation of airway safety of any newly synthesized neuromuscular blocking agents considered for clinical practice. PMID- 16306733 TI - Surfactants reduce platelet-bubble and platelet-platelet binding induced by in vitro air embolism. AB - BACKGROUND: The effect of gas bubbles on platelet behavior is poorly characterized. The authors assessed platelet-bubble and platelet-platelet binding in platelet-rich plasma in the presence and absence of bubbles and three surface active compounds. METHODS: Platelet-rich plasma was prepared from blood drawn from 16 volunteers. Experimental groups were surfactant alone, sparging (microbubble embolization) alone, sparging with surfactant, and neither sparging nor surfactant. The surfactants were Pluronic F-127 (Molecular Probes, Eugene, OR), Perftoran (OJSC SPC Perftoran, Moscow, Russia), and Dow Corning Antifoam 1510US (Dow Corning, Midland, MI). Videomicroscopy images of specimens drawn through rectangular glass microcapillaries on an inverted microscope and Coulter counter measurements were used to assess platelet-bubble and platelet-platelet binding, respectively, in calcium-free and recalcified samples. Histamine-induced and adenosine diphosphate-induced platelet-platelet binding were measured in unsparged samples. Differences between groups were considered significant for P < 0.05 using analysis of variance and the Bonferroni correction. RESULTS: Sixty to 100 platelets adhered to bubbles in sparged, surfactant-free samples. With sparging and surfactant, few platelets adhered to bubbles. Numbers of platelet singlets and multimers not adherent to bubbles were different (P < 0.05) compared both with unsparged samples and sparged samples without surfactant. No significant platelet-platelet binding occurred in uncalcified, sparged samples, although 20-30 platelets adhered to bubbles. Without sparging, histamine and adenosine diphosphate provoked platelet-platelet binding with and without surfactants present. CONCLUSIONS: Sparging causes platelets to bind to air bubbles and each other. Surfactants added before sparging attenuate platelet bubble and platelet-platelet binding. Surfactants may have a clinical role in attenuating gas embolism-induced platelet-bubble and platelet-platelet binding. PMID- 16306734 TI - Effect of labor epidural analgesia with and without fentanyl on infant breast feeding: a prospective, randomized, double-blind study. AB - BACKGROUND: The influence of labor epidural fentanyl on the neonate is controversial. The purpose of this study was to determine whether epidural fentanyl has an impact on breast-feeding. METHODS: Women who previously breast fed a child and who requested labor epidural analgesia were randomly assigned in a double-blinded manner to one of three groups: (1) no fentanyl group, (2) intermediate-dose fentanyl group (intent to administer between 1 and 150 microg epidural fentanyl), or (3) high-dose epidural fentanyl group (intent to administer > 150 microg epidural fentanyl). On postpartum day 1, the mother and a lactation consultant separately assessed whether the infant was experiencing difficulty breast-feeding, and a pediatrician assessed infant neurobehavior. All women were contacted 6 weeks postpartum to determine whether they were still breast-feeding. RESULTS: Sixty women were randomly assigned to receive no fentanyl, 59 were randomly assigned to receive an intermediate dose, and 58 were randomly assigned to receive high-dose fentanyl. On postpartum day 1, women who were randomly assigned to receive high-dose fentanyl reported difficulty breast feeding (n = 12, 21%) more often than women who were randomly assigned to receive an intermediate fentanyl dose (n = 6, 10%), or no fentanyl (n = 6, 10%), although this did not reach statistical significance (P = 0.09). There was also no significant difference among groups in breast-feeding difficulty based on the lactation consultant's evaluation (40% difficulty in each group; P = 1.0). Neurobehavior scores were lowest in the infants of women who were randomly assigned to receive more than 150 microg fentanyl (P = 0.03). At 6 weeks postpartum, more women who were randomly assigned to high-dose epidural fentanyl were not breast-feeding (n = 10, 17%) than women who were randomly assigned to receive either an intermediate fentanyl dose (n = 3, 5%) or no fentanyl (n = 1, 2%) (P = 0.005). CONCLUSIONS: Among women who breast-fed previously, those who were randomly assigned to receive high-dose labor epidural fentanyl were more likely to have stopped breast-feeding 6 weeks postpartum than woman who were randomly assigned to receive less fentanyl or no fentanyl. PMID- 16306735 TI - Frontal slab composite magnetic resonance neurography of the brachial plexus: implications for infraclavicular block approaches. AB - BACKGROUND: Magnetic resonance neurography (MRN) is an imaging method by which nerves can be selectively highlighted. Using commercial software, the authors explored a variety of approaches to develop a three-dimensional volume-rendered MRN image of the entire brachial plexus and used it to evaluate the accuracy of infraclavicular block approaches. METHODS: With institutional review board approval, MRN of the brachial plexus was performed in 10 volunteer subjects. MRN imaging was performed on a GE 1.5-tesla magnetic resonance scanner (General Electric Healthcare Technologies, Waukesha, WI) using a phased array torso coil. Coronal STIR and T1 oblique sagittal sequences of the brachial plexus were obtained. Multiple software programs were explored for enhanced display and manipulation of the composite magnetic resonance images. The authors developed a frontal slab composite approach that allows single-frame reconstruction of a three-dimensional volume-rendered image of the entire brachial plexus. Automatic segmentation was supplemented by manual segmentation in nearly all cases. For each of three infraclavicular approaches (posteriorly directed needle below midclavicle, infracoracoid, or caudomedial to coracoid), the targeting error was measured as the distance from the MRN plexus midpoint to the approach-targeted site. RESULTS: Composite frontal slabs (coronal views), which are single-frame three-dimensional volume renderings from image-enhanced two-dimensional frontal view projections of the underlying coronal slices, were created. The targeting errors (mean +/- SD) for the approaches-midclavicle, infracoracoid, caudomedial to coracoid-were 0.43 +/- 0.67, 0.99 +/- 1.22, and 0.65 +/- 1.14 cm, respectively. CONCLUSION: Image-processed three-dimensional volume-rendered MNR scans, which allow visualization of the entire brachial plexus within a single composite image, have educational value in illustrating the complexity and individual variation of the plexus. Suggestions for improved guidance during infraclavicular block procedures are presented. PMID- 16306736 TI - Comparison of morphine, ketorolac, and their combination for postoperative pain: results from a large, randomized, double-blind trial. AB - BACKGROUND: Meta-analyses report similar numbers needed to treat for nonsteroidal antiinflammatory drugs (NSAIDs) and opioids. Differences in baseline pain intensity among the studies from which these numbers needed to treat were derived may have confounded the results. NSAIDs have an opioid-sparing effect, but the importance of this effect is unclear. Therefore, the authors sought to compare the proportions of subjects who obtain pain relief with ketorolac versus morphine after surgery and to determine whether the opioid-sparing effect of an NSAID reduces the magnitude of opioid side effects. METHODS: The study was a double blind, randomized controlled trial. The authors randomly assigned 1,003 adult patients to receive 30 mg ketorolac or 0.1 mg/kg morphine intravenously. They calculated the proportion of subjects who achieved at least 50% reduction in pain intensity 30 min after analgesic administration. Further, so long as pain intensity 30 min after analgesic administration was 5 or more out of 10, patients received 2.5 mg morphine every 10 min until pain intensity was 4 or less out of 10. The authors assessed the presence of opioid-related side effects. RESULTS: Five hundred patients received morphine and 503 received ketorolac. Fifty percent of patients in the morphine group achieved pain relief, compared with 31% in the ketorolac group (difference, 19%; 95% confidence interval, 13-25%). The ketorolac morphine group required less morphine (difference, 6.5 mg; 95% confidence interval, -5.8 to -7.2) and had a lower incidence of side effects (difference, 11%; 95% confidence interval, 5-16%) than the morphine group. CONCLUSIONS: Opioids are more efficacious analgesics than NSAIDs, although historic data for these two drugs yield similar numbers needed to treat. Adding NSAIDs to the opioid treatment reduces morphine requirements and opioid-related side effects in the early postoperative period. PMID- 16306737 TI - Minimum local analgesic dose: effect of different volumes of intrathecal levobupivacaine in early labor. AB - BACKGROUND: This double-blind, randomized study was aimed at detecting the effect of three different volumes of intrathecal levobupivacaine on the minimum local analgesic dose in early labor. METHODS: Ninety-three nulliparous women requesting combined spinal-epidural analgesia, at more than 37 weeks gestation, with spontaneous onset of labor, cervical dilatation from 2 to 5 cm, were enrolled. Parturients received 10 ml (group 10), 5 ml (group 5), or 2.5 ml (group 2.5) of the spinal solution containing plain levobupivacaine diluted with 0.9% wt/vol saline to achieve the desired dose and volume at room temperature. A lumbar epidural catheter was then placed. The initial dose for each group was 2.0 mg, and the following doses were determined by the response of the previous patient using up-down sequential allocation. The authors required the test solution to achieve a visual analog pain score of 10 mm or less to be considered effective. The up-down sequences were analyzed using the Dixon and Massey formula and regression logistic model. RESULTS: The minimum local analgesic dose of spinal levobupivacaine in spontaneously laboring women was 1.35 mg (95% confidence interval, 1.25-1.45 mg) in group 10, 1.63 mg (95% confidence interval, 1.51-1.76 mg) in group 5, and 1.97 mg (95% confidence interval, 1.89-2.05 mg) in group 2.5. A unit change in volume increased the odds of an effective response multiplicatively by a factor of 1.8. CONCLUSIONS: Analgesia can be achieved using lower doses and higher volumes even in subarachnoid space. The important role of the volume should be considered not only in epidural but also in spinal analgesia. PMID- 16306738 TI - A new inguinal approach for the obturator nerve block: anatomical and randomized clinical studies. AB - BACKGROUND: Obturator nerve block is highly recommended for knee surgery in addition to a femoral nerve block. The main disadvantage of the classic approach at the pubic tubercle is low patient acceptance due to pain and discomfort. The authors hypothesized that the use of a new inguinal obturator nerve block technique would reduce pain and discomfort in patients. METHODS: The inguinal approach was simulated in five fresh cadavers. Injection of latex was performed in two cadavers. The location of the needle and the extent of latex solution were analyzed. Fifty patients scheduled to undergo arthroscopic knee surgery were randomly assigned to receive obturator nerve block using either the inguinal (n = 25) or the pubic tubercle approach (n = 25). RESULTS: In all cadavers, the needle was close to the obturator nerve branches, which were surrounded by the latex solution. In the clinical study, visual analog scale pain scores and discomfort of block placement were significantly lower in the inguinal group compared with the pubic tubercle group (P < 0.01). In the inguinal group, there was a significant decrease in block performance time (P < 0.05) and in bolus of propofol and fentanyl used for the procedure (P < 0.01). Twenty minutes after application of the block, adductor strength decrease, occurrence, and location of cutaneous distribution of the obturator nerve were not significantly different between the groups. The incidence of minor complications was significantly increased in the pubic tubercle group (P < 0.05). No major complications were observed. CONCLUSIONS: The new inguinal approach decreases patient discomfort and pain of block placement as well as the time and sedation and analgesics required for a similar quality of sensory and motor block compared with the pubic tubercle approach. PMID- 16306739 TI - Ephedrine blocks rat sciatic nerve in vivo and sodium channels in vitro. AB - BACKGROUND: The sympathomimetic drug ephedrine has been used intrathecally as the sole local anesthetic for labor and delivery. Because ephedrine may be a useful adjuvant to local anesthetics, the authors investigated the local anesthetic properties of ephedrine in a rat sciatic nerve block model and the underlying mechanism in cultured cells stably expressing Na channels. METHODS: After approval of the animal protocol, the sciatic nerves of anesthetized rats were exposed by lateral incision of the thighs, 0.2 ml ephedrine at 0.25, 1, 2.5, or 5% and/or bupivacaine at 0.125% was injected, and the wound was closed. Motor and sensory/nociceptive functions were evaluated by the force achieved by pushing against a balance and the reaction to pinch, respectively. The whole cell configuration of the patch clamp technique was used to record Na currents from human embryonal kidney cells stably transfected with Nav1.4 channels. RESULTS: The nociception blockade was significantly longer than the motor blockade at test doses of 2.5 and 5% of ephedrine, or when 1% ephedrine was combined with 0.125% bupivacaine (analysis of variance with repeated measures, P < 0.001, n = 8/group). In vitro, the 50% inhibitory concentrations of ephedrine at -150 and 60 mV were 1,043 +/- 70 and 473 +/- 13 mum, respectively. High-frequency stimulation revealed a use-dependent block of 18%, similar to most local anesthetics. CONCLUSIONS: Because ephedrine's properties are at least partly due to Na channel blockade, detailed histopathologic investigations are justified to determine the potential of ephedrine as an adjuvant to clinically used local anesthetics. PMID- 16306740 TI - Spinal muscarinic and nicotinic subtypes activated by clonidine in postincisional pain. AB - BACKGROUND: A recent model of acute incisional pain has been characterized that strongly parallels the postoperative period in patients experiencing evoked pain. In that setting, abundant literature has revealed antihypersensitive effects produced by intrathecally administered alpha2-adrenergic receptor agonists, such as clonidine, in both animals and humans. Recent reports have suggested an obligatory role of spinal acetylcholine receptors in the analgesic action of intrathecal clonidine. The authors sought to determine the involvement of spinal muscarinic and nicotinic receptor subpopulations in the antihypersensitivity effect of intrathecal clonidine in a rodent model for human postoperative pain. METHODS: After intrathecal catheterization, rats underwent superficial plantar incision. Clonidine or a combination of clonidine and muscarinic receptor subtype antagonists (M1, M2, M3, and M4) or nicotinic receptor subtype antagonists (alpha4beta2 and alpha7) were intrathecally administered, and withdrawal thresholds to mechanical stimuli were examined. RESULTS: Spinal clonidine maximally reduced hypersensitivity adjacent to the wound 30 min after its injection. When animals were intrathecally pretreated with the M1 muscarinic antagonist toxin MT-7, the M3 muscarinic antagonist 4-diphenylacetoxy-N methylpiperidine, and the M4 muscarinic antagonist toxin MT-3, clonidine lost its antihypersensitive action. When animals were intrathecally pretreated with the alpha4beta2 nicotinic receptor antagonist dihydro-beta-erythroidine, but not with the alpha7 nicotinic receptor antagonist methyllycaconitine, the antihypersensitivity action of clonidine was abolished. CONCLUSIONS: These data indicate for the first time that the clonidine-induced increase in punctuate mechanical threshold is mediated via the activation of all but M2 muscarinic receptor subtypes, and via the activation of alpha4beta2 but not alpha7 nicotinic receptor subtypes in a rodent model for human postoperative pain. PMID- 16306741 TI - Bayesian prediction bounds and comparisons of operating room times even for procedures with few or no historic data. AB - BACKGROUND: Lower prediction bounds (e.g., for fasting), upper prediction bounds (e.g., to schedule delays between sequential surgeons), comparisons of operating room (OR) times (e.g., when sequencing cases among ORs), and quantification of case uncertainty (e.g., for sequencing a surgeon's list of cases) can be done accurately for combinations of surgeon and scheduled procedure(s) by using historic OR times. The authors propose that when there are few or no historic data, the predictive distribution of the OR time of a future case be centered at the scheduled OR time, and its proportional uncertainty be based on that of other surgeons and procedures. When there are a moderate or large number of historic data, the historic data alone are used in the prediction. When there are a small number of historic data, a weighted combination is used. METHODS: This Bayesian method was tested with all 65,661 cases from a hospital. RESULTS: Bayesian prediction bounds were accurate to within 2% (e.g., the 5% lower bounds exceeded 4.9% of the actual OR times). The predicted probability of one case taking longer than another was estimated to within 0.7%. When sequencing a surgeon's list of cases to reduce patient waiting past scheduled start times, both the scheduled OR time and the variability in historic OR times should be used together when assessing which cases should be done first. CONCLUSIONS: The authors validated a practical way to calculate prediction bounds and compare the OR times of all cases, even those with few or no historic data for the surgeon and the scheduled procedure(s). PMID- 16306742 TI - Sleep, anesthesiology, and the neurobiology of arousal state control. AB - Sleep, like breathing, is a biologic rhythm that is actively generated by the brain. Neuronal networks that have evolved to regulate naturally occurring sleep preferentially modulate traits that define states of sedation and anesthesia. Sleep is temporally organized into distinct stages that are characterized by a unique constellation of physiologic and behavioral traits. Sleep and anesthetic susceptibility are genetically modulated, heritable phenotypes. This review considers 40 yr of research regarding the cellular and molecular mechanisms contributing to arousal state control. Clinical and preclinical data have debunked and supplanted the primitive view that sleep need is a weakness. Sleep deprivation and restriction diminish vigilance, alter neuroendocrine control, and negatively impact immune function. There is overwhelming support for the view that decrements in vigilance can negatively impact performance. Advances in neuroscience provide a foundation for the sea change in public and legal perspectives that now regard a sleep-deprived individual as impaired. PMID- 16306743 TI - Does multimodal analgesia with acetaminophen, nonsteroidal antiinflammatory drugs, or selective cyclooxygenase-2 inhibitors and patient-controlled analgesia morphine offer advantages over morphine alone? Meta-analyses of randomized trials. AB - The authors analyzed data from 52 randomized placebo-controlled trials (4,893 adults) testing acetaminophen, nonsteroidal antiinflammatory drugs, or selective cyclooxygenase-2 inhibitors given in conjunction with morphine after surgery. The median of the average 24-h morphine consumption in controls was 49 mg (range, 15 117 mg); it was significantly decreased with all regimens by 15-55%. There was evidence of a reduction in pain intensity at 24 h (1 cm on the 0- to 10-cm visual analog scale) only with nonsteroidal antiinflammatory drugs. Nonsteroidal antiinflammatory drugs also significantly reduced the incidence of nausea/vomiting from 28.8% to 22.0% (number needed to treat, 15) and of sedation from 15.4% to 12.7% (number needed to treat, 37) but increased the risk of severe bleeding from 0% to 1.7% (number needed to harm, 59). Selective cyclooxygenase-2 inhibitors increased the risk of renal failure in cardiac patients from 0% to 1.4% (number needed to harm, 73). A decrease in morphine consumption is not a good indicator of the usefulness of a supplemental analgesic. There is evidence that the combination of nonsteroidal antiinflammatory drugs with patient controlled analgesia morphine offers some advantages over morphine alone. PMID- 16306744 TI - Developing social capital in the operating room: the use of population-based techniques. PMID- 16306745 TI - Pulsed radiofrequency: a neurobiologic and clinical reality. PMID- 16306746 TI - A comment on the history of the pulsed radiofrequency technique for pain therapy. PMID- 16306748 TI - Pulsed radiofrequency. PMID- 16306749 TI - Graphical display of data could reveal errors in statistical tests. PMID- 16306750 TI - Some points regarding anesthesia for patients with congenital long QT syndrome. PMID- 16306753 TI - Volatile anesthetics and the long QT syndrome. PMID- 16306755 TI - On the origin of critical care units: a clarification. PMID- 16306757 TI - Extended-release epidural morphine formulation data far from clear. PMID- 16306759 TI - Anesthetics and memory: on memory at the cognitive and cellular levels. PMID- 16306760 TI - Regular clinical use bispectral index monitoring may result in lighter depth of anesthesia as reflected in average higher bispectral index values. PMID- 16306761 TI - XXIVth Annual Meeting of the European Malignant Hyperthermia Group. PMID- 16306762 TI - 43rd Western Anesthesia Residents Conference. PMID- 16306764 TI - Association of polymorphisms in the interleukin-18 gene in patients with Crohn's disease depending on the CARD15/NOD2 genotype. AB - An increased expression of interleukin-18 (IL-18), a proinflammatory cytokine inducing interferon-gamma, has been found in Crohn's disease (CD). In the IL-18 gene, several partly functional relevant polymorphisms are known. This study sought to investigate associations of IL-18 polymorphisms in inflammatory bowel disease and CD according to CARD15/NOD2 mutation status and clinical phenotypes. METHODS: The IL-18 polymorphisms -607, -137, and the third position of codon 35 (c35/3) were genotyped in 210 patients with CD, 140 patients with ulcerative colitis, and 265 healthy controls using polymerase chain reaction and restriction fragment length polymorphism analysis. RESULTS: Frequencies of alleles and genotypes of the 3 polymorphisms and of the respective haplotypes and diplotypes displayed no significant differences between the whole groups of patients with CD and ulcerative colitis, respectively, compared with the controls. After stratification of patients with CD for CARD15/NOD2 status, significant associations of genotypes -137 CC (P = 0.018) and c35/3 CC (P = 0.010) and of the diplotype 2-2 (P = 0.018) were found in cases carrying CARD15/NOD2 mutations. Associations of genotypes -137 GG (P = 0.015) and c35/3 AA (P = 0.030) with colonic disease only in cases positive for CARD15/NOD2 mutations and of the genotype -607 AA (P = 0.007) with fistulas in cases negative for CARD15/NOD2 mutations were observed. CONCLUSIONS: In this study, significant differences of several genotypes and diplotypes within the IL-18 gene in CD depending on CARD15/NOD2 status have been found. In context with an increased expression of IL 18 in CD, it remains to be shown whether the expression of IL-18 is influenced by CARD15/NOD2 mutation status. PMID- 16306765 TI - Association between IL-18 gene promoter polymorphisms and inflammatory bowel disease in a Japanese population. AB - BACKGROUND: Interleukin-18 (IL-18) is a pleiotropic cytokine that induces the production of interferon (IFN)-gamma and also to regulate Th2 cytokines. Recently, association studies between IL-18 gene promoter polymorphisms and several Th1- or Th2-mediated inflammatory diseases were reported. In inflammatory bowel diseases (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), recent evidence suggests that IL-18 is involved in the pathogenesis. METHODS: Using DNA direct sequencing, we investigated IL-18 gene promoter polymorphisms at -607C/A and -137G/C. Allele, genotype, and haplotype frequencies were determined in 210 Japanese patients with UC, 205 patients with CD, and 212 controls. RESULTS: In UC, the -137C allele frequency was significantly higher in the proctitis-type patients than in controls (Pc = 0.0068). The -137 genotype frequency was also significantly different in the proctitis-type patients than in controls (Pc = 0.032). No other allele and genotype frequencies were significantly associated with UC after Bonferroni correction. Furthermore, the frequency of haplotype 2 (-607A, -137C), which had a lower promoter activity and IFN-gamma mRNA level than the other haplotypes as previously reported, was significantly higher in the proctitis-type patients than in controls (Pc = 0.01). In CD, we could not find any significant differences. CONCLUSIONS: IL-18 gene promoter polymorphisms may not be associated with disease susceptibility but related to the extent of disease in UC. PMID- 16306766 TI - T helper 1-inducing property of IL-27/WSX-1 signaling is required for the induction of experimental colitis. AB - BACKGROUND: WSX-1, a component of the interleukin (IL)-27 receptor, is a novel class I cytokine receptor with homology to the IL-12 receptor beta2 chain. Initially, WSX-1 signaling was reported to play an important role in the promotion of T helper-1 responses, but recent reports have revealed an anti inflammatory property in WSX-1 signaling. In the present study, we investigated the role of IL-27/WSX-1 signaling in a murine colitis model, dextran sulfate sodium (DSS) colitis, by using WSX-1 knockout (KO) mice. METHODS: First, we observed whether WSX-1 KO mice developed colitis spontaneously. Second, we induced DSS colitis in WSX-1 KO and wild-type (WT) mice. RESULTS: WSX-1 KO mice were observed not to develop colitis spontaneously. The severity of DSS colitis was decreased in WSX-1 KO mice in comparison with WT mice in association with a reduced production of interferon-gamma, IL-6, and tumor necrosis factor-alpha by lamina propria mononuclear cells from WSX-1 KO mice and the absence of T-bet expression in the colon from WSX-1 KO mice. CONCLUSIONS: This study revealed the inflammatory property of IL-27/WSX-1 signaling in intestinal inflammation. As a result, IL-27/WSX-1 signal pathway may thus be a promising candidate for the therapeutic intervention of human inflammatory bowel diseases such as Crohn's disease and ulcerative colitis. PMID- 16306767 TI - Regulatory role of the pituitary-adrenal axis in experimental colitis: effect of adrenalectomy on the clinical course and the TH1/TH2 immune profile. AB - BACKGROUND: The hypothalamic-pituitary-adrenal (HPA) axis plays an important role in modulating immune reactions in inflammatory bowel disease. Our aim was to assess the role of the HPA axis in the pathogenesis of immunomediated colitis in mice. METHODS: Trinitrobenzene sulfonic acid (TNBS) colitis was induced in Balb/c mice. Sham operation (sham+TNBS) or bilateral adrenalectomy (Adex+TNBS) was performed 3 days later. Control groups underwent adrenalectomy without colitis induction (Adex) or were untreated [naive mice (Naive)]. Mice were monitored for survival, weight loss, and macroscopic and microscopic scores of colitis. FACS analysis of CD4, CD8, natural killer T lymphocytes, and serum levels of adrenocorticotropic hormone (ACTH), corticosterone (CS), interferon-gamma (IFN gamma), interleukin-10 (IL-10), and IL-1beta were measured. Production of prostaglandin E2 (PGE2) and binding capacity to glucocorticoid receptor (GR) in colonic mucosa were also assessed. RESULTS: By day 7 following induction of colitis there was a marked increase in ACTH and CS levels in the colitis as compared with the control group (86 +/- 6.5 pg/mL and 16 +/- 1.9 pg/mL, and 23.3 +/- 2 pg/mL and 2.8 +/- 0.8 pg/mL, respectively). There was a decrease in ACTH and CS levels by day 28 in the colitis group, but the levels were still significantly higher than the levels in controls. Adrenalectomy markedly exacerbated colitis. The macroscopic and microscopic scores increased from 2.79 +/- 0.03 and 2.0 +/- 0.1 in the sham+TNBS group to 3.3 +/- 0.3 and 3.2 +/- 0.3 in the Adex+TNBS group. Survival and weight loss correlated with these differences. A significant increase in IL-10, IFN-gamma, and PGE2 was noted in the Adex+TNBS group compared with the sham+TNBS group. Splenic CD4 lymphocytes decreased in the sham+TNBS and Adex+TNBS groups as compared with control groups (Adex and naive). The CD8/CD4 ratio was significantly higher in the Adex+TNBS compared with the sham+TNBS group. Colitis also caused a significant decrease in the specific binding capacity of labeled dexamethasone to colonic mucosa. CONCLUSIONS: TNBS induced colitis activated the HPA axis and reduced the sensitivity of the inflamed mucosa to circulating glucocorticoids. Adrenalectomy markedly exacerbated TNBS-induced colitis. The effect was associated with changes in the peripheral CD8/CD4 ratio and with a TH1 cytokine shift. Our results suggest that adrenocortical hormones play an important role in the regulation of the immune system in experimental colitis. PMID- 16306768 TI - Piroxicam treatment of IL-10-deficient mice enhances colonic epithelial apoptosis and mucosal exposure to intestinal bacteria. AB - Treatment with the nonsteroidal anti-inflammatory drugs piroxicam or sulindac was recently shown to accelerate the development of colitis in interleukin (IL)-10 deficient (IL-10) mice. Although NSAIDs have been hypothesized to decrease the barrier function of the intestinal epithelium, the mechanism by which this accelerates colitis in IL-10 mice is not well understood. In this study, the effects of piroxicam on the colonic mucosa of IL-10 C57BL/6 mice were evaluated histologically. The effect of piroxicam on intestinal epithelial cells in vitro was assessed using colorimetric and fluorescent assays for cell viability and apoptotic cell death. Interactions of intestinal bacteria with the colonic mucosa were evaluated by rRNA-directed fluorescence in situ hybridization. In vivo treatment of C57BL/6 IL-10 mice with oral piroxicam markedly enhanced apoptosis of colonic epithelium and resulted in focal erosion of the mucosal surface, enhanced bacterial adhesion and invasion, and accelerated the development of colitis. In vitro, piroxicam induced apoptosis of CT26 murine intestinal epithelial cells in a dose-dependent fashion. Piroxicam-induced apoptosis of CT26 cells could not be prevented by addition of exogenous IL-10; however, IL-10 did significantly enhance their rate of proliferation. Thus, exposure to piroxicam enhances intestinal epithelial apoptosis both in vitro and in vivo and facilitates adhesion and invasion of intestinal bacteria into mucosal tissues in vivo. The role of IL-10 in this process requires further study. These studies support the hypothesis that increased exposure of mucosal cells to intestinal bacteria may lead to development of intestinal inflammation in IL-10 or other genetically susceptible individuals. PMID- 16306769 TI - Increased expression of MIP-3alpha/CCL20 in peripheral blood mononuclear cells from patients with ulcerative colitis and its down-regulation by sulfasalazine and glucocorticoid treatment. AB - CCL20 expression is known to increase in the mucosal tissues of inflammatory bowel diseases (IBDs). Moreover, the discovery of Nod2 as the IBD1 susceptibility gene has underscored the significance of blood mononuclear cells in IBD pathogenesis. METHODS: This study addresses whether CCL20 expression is similarly altered in peripheral blood mononuclear cells (PBMCs) of patients with ulcerative colitis (UC), a major type of IBD in Korea. RESULTS: Expression of CCL20 was significantly up-regulated in the PBMCs of patients with UC compared with those of normal healthy controls. Interestingly, untreated UC groups expressed higher levels of CCL20 mRNA than either treated UC or normal control groups, suggesting that CCL20 could be modulated by anti-inflammatory drugs. Accordingly, a strong association between CCL20 levels and disease activity index was observed. Supporting these findings, results from a 3-month follow-up study revealed that the UC groups treated with 5-aminosalicylic acid and glucocorticoid exhibited dramatic decreases of CCL20 mRNA in PBMCs, accompanied by ameliorated disease states. Moreover, tumor necrosis factor-alpha- or interleukin-1beta-induced CCL20 secretion was greatly diminished by 5-aminosalicylic acid and/or glucocorticoid treatment of human intestinal epithelial HT-29 cells. Of note, CCR6 cell populations were significantly reduced in the blood of severe patients with UC compared with normal controls, whereas no significant changes in CCR6 cell populations were observed in the blood of patients with mild UC or acute colitis. CONCLUSIONS: Collectively, these findings suggest that CCL20 expression in blood mononuclear cells is associated with altered immune and inflammatory responses in patients with UC. PMID- 16306770 TI - Acute pancreatitis in patients with Crohn's disease: clinical features and outcomes. AB - BACKGROUND: Acute pancreatitis has occasionally been associated with Crohn's disease (CD), but whether a causal association exists remains unclear. We sought to determine the frequency of etiologies in a consecutive series of patients with CD with acute pancreatitis. METHODS: A centralized diagnostic index was used to identify all patients with CD with acute pancreatitis that were evaluated at Mayo Clinic Rochester between 1976 and 2001. Both diagnoses were made or confirmed at our institution. Records were abstracted for demographics, presenting symptoms, diagnostic tests, risk factors of pancreatitis, treatment, and follow-up. RESULTS: Forty-eight patients with CD with pancreatitis were identified. The median age at diagnosis of acute pancreatitis was 47 years (range, 31-91 yr). Forty-six (96%) met biochemical criteria for acute pancreatitis. The most sensitive radiographic tests were abdominal computed tomography (70%) and abdominal ultrasound (46%). The etiology of pancreatitis was considered to be gallstones (21%), significant alcohol intake (15%), use of purine analogs (13%), duodenal Crohn's involvement (12%), postendoscopic retrograde cholangiopancreatography complications (10%), postoperative complications (12%), use of other medications (4%) and idiopathic (8%). The median length of hospitalization was 7 days (range, 0-40 d). Ten patients (21%) had recurrence of acute pancreatitis. Three patients (6%) were subsequently diagnosed with pancreatic cancer. CONCLUSIONS: A definite etiology could be identified in most patients with CD with acute pancreatitis. Gallstones and alcohol accounted for more than one third of cases, whereas CD, either because of duodenal involvement or medications used for its treatment, accounted for more than one quarter of the cases. A small proportion of patients remained idiopathic. PMID- 16306771 TI - Comparison of 4 neutrophil-derived proteins in feces as indicators of disease activity in ulcerative colitis. AB - BACKGROUND: To evaluate the diagnostic use of fecal concentrations of lactoferrin (Lf), calprotectin (Cal), polymorphonuclear neutrophil-elastase (PMN-e), and lysozyme (Lys) as indicators of disease activity in patients with active and inactive ulcerative colitis (UC). METHODS: A total of 76 fecal specimens were collected from 31 patients with UC in times of active and inactive status of disease. Disease activity was determined with the colitis activity index (CAI; Rachmilewitz index), which includes a combination of laboratory parameters and clinical symptoms, with a score of at least 6 indicating active disease. Fecal Lf, Cal, PMN-e, and Lys were measured and reported as micrograms per milliliter feces. Levels of more than 7.25, more than 6.00, at least 0.062, and at least 0.6 for Lf, Cal, PMN-e, and Lys, respectively, were considered elevated as specified by the manufacturers. RESULTS: Based on the CAI classification, 25 of the samples were from patients with active disease status and 51 were from patients with inactive status. Lf, PMN-e, and Cal but not Lys showed increased levels in samples from patients in active disease compared with those in remission (median for Lf: 28.12 +/- 110.86 versus 179.54 +/- 334.09, P < 0.001; median for Cal: 15.13 +/- 30.27 versus 116.23 +/- 182.29, P < 0.001; median for PMN-e: 0.21 +/- 0.44 versus 1.02 +/- 0.89, P < 0.001; median for Lys: 1.54 +/- 2.39 versus 3.75 +/- 5.39, P > 0.05). All 4 parameters correlated with the CAI (Lf: r = 0.441, P < 0.001; Cal: r = 0.505, P < 0.001; PMN-e: r = 0.604, P < 0.001; Lys: r = 0.295, P < 0.05). Introducing a composite index based on Lf, Cal, and PMN-e, the specificity was 72.5% and the sensitivity 88% compared with the CAI. CONCLUSIONS: Among the neutrophil-derived proteins in feces, PMN-e, Cal, and Lf represent useful markers of disease activity in patients with UC. Using all 3 markers in a composite index may be an additional noninvasive tool for the management of ambulant patients with UC. PMID- 16306772 TI - Preliminary observations of oral nicotine therapy for inflammatory bowel disease: an open-label phase I-II study of tolerance. AB - BACKGROUND: Transdermal nicotine provides benefit in active ulcerative colitis but is often associated with adverse events (AEs). An oral formulation has been developed to minimize AEs. This study was undertaken to make initial observations on the safety and tolerance of oral nicotine therapy in inflammatory bowel disease; the effect on disease activity was also noted. METHODS: Twenty-six patients with ulcerative colitis, 11 with active disease, and 5 patients with Crohn's colitis (2 with active disease) were given oral nicotine in 3-mg capsules, gradually increasing the dose to the maximum tolerated. AEs were recorded, concomitant prednisolone and/or azathioprine were reduced where possible, and disease activity was reassessed at the end of nicotine treatment. RESULTS: Patients were followed for up to 12 months. Twenty-nine of 31 could tolerate at least 6 mg of nicotine each day, and 5 patients tolerated at least 18 mg daily. Twenty-four patients had nicotine-related nonserious AEs; over one half occurred during the period of dose escalation, but 7 discontinued treatment because of them. Six of the 13 patients with active disease became asymptomatic, whereas 3 patients in remission developed active symptoms; 11 patients reduced their concomitant medication. CONCLUSIONS: Oral nicotine is a safe potential treatment of inflammatory bowel disease, but there is considerable variation in tolerance. PMID- 16306773 TI - Ubiquitin protein modification and signal transduction: implications for inflammatory bowel diseases. AB - A dysregulated immune response to luminal antigen(s) is associated with the development of inflammatory bowel diseases (IBDs). A complex network of inflammatory and immune mediators released by immune and nonimmune cells participate in the physiopathology of IBD. At the molecular level, events leading to the improper use of the signaling grid are likely responsible for the dysregulated activation of various transcription factors and subsequent induction of inflammatory genes. The posttranslational modification of signaling proteins by the ubiquitin system is a critical event in activation or repression of transcription factors. Two important transcriptional pathways in which ubiquitin is central are the nuclear factor-kappaB and hypoxia inducible factor-1 (HIF-1) pathways, both of which are important components of intestinal homeostasis. In this review, we discuss the role of ubiquitin modification in relation to nuclear factor-kappaB and HIF-1 signaling and consider its impact on intestinal inflammation. A greater understanding of posttranslational ubiquitin modification may lead to the identification of new therapeutic opportunities for the treatment of IBD. PMID- 16306774 TI - Bone loss in Crohn's disease: exercise as a potential countermeasure. AB - Crohn's disease (CD) is associated with a number of secondary conditions including osteoporosis, which increases the risk of bone fracture. The cause of metabolic bone disease in this population is believed to be multifactorial and may include the disease itself and associated inflammation, high-dose corticosteroid use, weight loss and malabsorption, a lack of exercise and physical activity, and an underlying genetic predisposition to bone loss. Reduced bone mineral density has been reported in between 5% to 80% of CD sufferers, although it is generally believed that approximately 40% of patients suffer from osteopenia and 15% from osteoporosis. Recent studies suggest a small but significantly increased risk of fracture compared with healthy controls and, perhaps, sufferers of other gastrointestinal disorders such as ulcerative colitis. The role of physical activity and exercise in the prevention and treatment of CD-related bone loss has received little attention, despite the benefits of specific exercises being well documented in healthy populations. This article reviews the prevalence of and risk factors for low bone mass in CD patients and examines various treatments for osteoporosis in these patients, with a particular focus on physical activity. PMID- 16306775 TI - TPMT genotype screening for patients about to begin azathioprine treatment--a look at costs and potential benefits. PMID- 16306776 TI - Immunomodulators and the need for surgery in Crohn's disease: insufficient evidence to convict. PMID- 16306777 TI - Successful infliximab therapy for a duodenal stricture caused by Crohn's disease. PMID- 16306778 TI - Debate on the lack of evidence of Mycobacterium avium subsp. paratuberculosis in Crohn's disease. PMID- 16306779 TI - Lymphoma of the female genital tract: current status. AB - SUMMARY: : Primary lymphomas affecting the female reproductive system are uncommon but often pose a diagnostic challenge if their existence is not suspected. This article reviews the pathological and clinical features of lymphomas occurring in various sites in the female genital tract including the vulva, vagina, cervix, endometrium, fallopian tubes, and ovaries. Using the recent World Health Organization classification, the various types of lymphomas are identified as separate diseases and not as morphological variations of the same disease. The immunophenotypic and cytogenetics features of the major lymphomas are summarized. The incidence, presenting symptoms, gross and microscopic features, major differential diagnostic considerations, response to therapy, and expected outcome are discussed. Using published data on patient outcome, the International Federation of Obstetricians and Gynecologists and Ann Arbor staging systems are compared for their predictive value, and the difficulty in assigning primary and secondary status in extranodal lymphomas is emphasized. The observed differences in the behavior of some lymphomas in gynecological sites compared with their usual nodal location are presented. Finally, the possible etiology of these conditions is discussed in light of the emerging paradigm of mucosa-associated lymphoid tissue lymphomas. PMID- 16306780 TI - Vulvar squamous cell carcinoma is a multifactorial disease following two separate and independent pathways. AB - Two separate pathways leading to vulvar carcinoma have been suggested. First, a human papillomavirus (HPV)-dependent pathway, in which premalignant stages of vulvar cancer are the classic vulvar intraepithelial neoplasia (VIN) lesions. Second, an HPV-independent pathway, associated with differentiated VIN III lesions and/or lichen sclerosus. To obtain insight into the mechanisms underlying these pathways, we determined the relationship between HPV DNA and the expression of p14(ARF) and p16(INK4A) in non- and (pre)malignant vulvar lesions. Seventy three archival samples of non- and (pre)neoplastic vulvar lesions were selected and tested for hr-HPV DNA using a broad-spectrum HPV detection/genotyping assay (SPF(10)-LiPA) and the expression of p14(ARF) and p16(INK4A). The prevalence of HPV increased with the severity of the classic VIN lesions; in VIN I no hr-HPV was detected, in VIN II 43%, and in VIN III 71% of the samples were hr-HPV positive. Roughly the same was true for the expression of p14(ARF) and p16(INK4A). The simultaneous expression of p14(ARF) and p16(INK4A) was highly associated with the presence of hr-HPV DNA. Hr-HPV was detected in only a single case of the differentiated VIN III lesions, whereas no expression of p14(ARF) was found and 16(INK4A) was present in only two cases. All 16 samples of vulvar cancer were hr-HPV DNA- negative, although in respectively 63% and 25%, p14(ARF) and p16(INK4A) was expressed. No relation was found between hr-HPV and the expression of p14(ARF) and p16(INK4A) in the 20 nonneoplastic vulvar lesions. Our results provide further evidence that vulvar squamous cell carcinoma is a multifactorial disease that develops from two different pathways. First, an HPV dependent pathway with a remarkable resemblance to CIN lesions and cervical carcinoma and second, an HPV-independent pathway in which differentiated VIN III lesions that are hr-HPV-negative may be precursors. PMID- 16306781 TI - Galectin-3 and CD1a-positive dendritic cells are involved in the development of an invasive phenotype in vulvar squamous lesions. AB - In this study, the expression patterns of Galectin-3 (Gal-3) and the frequency of infiltrating CD1a positive dendritic cells (DCs) were determined in 82 cases of vulvar tissues, consisting of normal squamous epithelia (NE, N = 10), vulvar condylomas (VC, N = 24), high grade vulvar intraepithelial neoplasias (HG-VIN, N = 26) of common type, and invasive keratinizing squamous cell carcinomas (SCC, N = 22) by a standard immunohistochemical method using monoclonal antibodies to investigate their differential expression in vulvar squamous dysplasia and infiltrating carcinomas with an emphasis on neoplastic transformation and progression. Gal-3 expression was cytoplasmic, nuclear or membranous in NE, VCs, and HG-VINs, with negative or weak and occasionally moderate reactivities. In SCCs, exclusively cytoplasmic staining patterns with moderate or strong reactivity in 59% of cases were observed (p < 0.0001, chi-square test); Gal-3 expression was not related with stage, grade, and recurrence. The frequency of CD1a positive DCs increased from NE and VCs to highest numbers in HG-VINs, was lowest in SCCs (p < 0.0001, ANOVA), and was not related with stage and grade, but with recurrence in SCCs (p = 0,048, t-test). This study indicates that qualitative and quantitative changes of Gal-3 immunoexpression and infiltration by CD1a positive DCs in vulvar NE, VCs, and HG-VIN lesions, respectively, compared with SCCs play a role in the development of an infiltrative phenotype, and may provide adjunctive criteria in the diagnosis of invasion of vulvar squamous epithelia. PMID- 16306782 TI - Screening for cervical cancer in high-risk populations: DNA pap test or Hybrid Capture II test alone? AB - This study was designed to evaluate whether Hybrid Capture II (HC2) test alone refer women to colposcopy as appropriately as DNA Papanicolaou (Pap) test, in the context of a high-risk group of women using the recently validated DNACitoliq LBC system. Women with suspected cervical disease were included in this cross sectional study at a tertiary center in Sao Paulo, Brazil, for further workup. All women had cervical material collected for LBC and HC2 for high-risk human papillomavirus (hrHPV)-DNA test. Irrespective of cytology and HC2 results, colposcopy, and cervical biopsy when applicable, was systematically performed. All tests were performed blindly. Sensitivity, specificity, positive and negative predictive values, and overall accuracy of both methods were computed in relation to histology. A total of 1,080 women were included: 36.4% (393/1080) had ACUS+, 10.2% (110/1080) were high-grade squamous intraepithelial lesions (HSIL) or cancer. Mean age was 33.5 years. All women underwent colposcopy, and cervical biopsies were performed in 38.4% (415/1080): 33% (137/415) of the biopsies were negative, 14.4% (155/415) were low-grade squamous intraepithelial lesions (LSIL), 10.7% (116/415) were HSIL, and 0.6% (7/415) were cancer. HC2 sensitivity to diagnose biopsy-proven HSIL was 100%. Because all HSIL cases had a positive HC2 test, sensitivity could not be improved by adding LBC. Specificity and positive and negative predictive values of DNA Pap were not significantly different from HC2 test alone when considering LSIL+ histology as "gold standard" and HSIL+ histology. As a screening strategy for women with high-risk for cervical cancer, DNA Pap test does not seem to add substantially to HC2 alone in terms of appropriately referring to colposcopy. PMID- 16306783 TI - Herpes simplex virus and human papillomavirus infection in cervical disease in Argentine women. AB - The aim of the present study was to determine that prevalence of herpes simplex virus (HSV) type 1 and 2 in cervical samples from Argentine women and to assess the role of HSV-2 in cervical cancer. A sample of 79 normal and 200 neoplastic cervical tissues (35 invasive cervical carcinomas, 75 high-grade squamous intraepithelial lesions, 79 low-grade squamous intraepithelial lesions and 11 abnormal squamous cells of undermined significance) was analyzed for herpes simplex and human papillomavirus DNA using the polymerase chain reaction method. Viral genotyping was performed by single strand conformation polymorphisms and restriction fragment length polymorphisms. The overall prevalence of HSV was 21.5% in controls and 29% in cases. Among women with normal cytology, herpes simplex prevalence in HPV positive (20.8%) women was approximately the same as in negative (21.8%) women. HPV- and age- adjusted ORs of high-grade squamous intraepithelial lesions and invasive cervical carcinomas for HSV-2 were 1.4 (p = 0.6) and 1.6 (p = 0.5), respectively. The obtained results indicated that herpes simplex virus may not be involved in cervical cancer development. Future investigations are needed to provided conclusive evidence on the role of this pathogen in cervical cancer. PMID- 16306784 TI - Primary langerhans cell histiocytosis of the vulva. AB - Langerhans cell histiocytosis (LCH) of the female genital tract is very rare. A review of the literature revealed that only 12 cases of primary vulvar LCH have previously been published. We describe an additional case of LCH in which the disease was confined to the vulva. A 64-year-old woman was admitted to our hospital with a nodular mass on her left labium minus, and complete surgical excision was performed. On histological and immunohistochemical examination, the tumor fulfilled the criteria of LCH. A metastatic workup did not reveal any evidence of disease beyond the vulva. The patient received local radiotherapy, and 22 months later she is in excellent condition without local recurrence or metastatic disease. PMID- 16306785 TI - Association of low-grade endometrioid carcinoma of the uterus and ovary with undifferentiated carcinoma: a new type of dedifferentiated carcinoma? AB - Low-grade endometrioid carcinomas, either of the endometrium or the ovaries, usually have an excellent prognosis. The association of this type of tumor with undifferentiated carcinoma is rare. In this study, we present the clinicopathologic features of 25 such cases. The age of the patients ranged from 30 to 82 years (median, 51 years). At presentation, the patients had either vaginal bleeding or pelvic pain. The endometrioid carcinoma involved the endometrium in 14 cases, the endometrium and 1 or both ovaries in 9 cases, and the ovaries in 2 cases. Undifferentiated carcinoma associated with low-grade endometrioid carcinoma was found at presentation in 19 grade 1 or 2 endometrioid carcinomas: 15 in the endometrium and 5 in the ovary. In one of these cases, undifferentiated carcinoma was found in the endometrium and the ovary. Undifferentiated carcinoma was found after resection of low-grade endometrioid carcinoma in six cases, involving the retroperitoneum, pelvis, vagina, or liver. The undifferentiated carcinoma was composed exclusively of diffuse sheets and solid nests of epithelial cells in l0 cases. Epithelial cells with isolated foci of keratinization were seen in nine cases and rhabdoid cells in a myxoid background in six cases. Twenty-four patients were treated with total abdominal hysterectomy and with bilateral salpingo-oophorectomy. Twenty-two patients received additional therapy as follows: chemotherapy (), radiotherapy (), and tamoxifen (). Follow-up showed that 15 patients died of disease in 1 to 60 months (median, 6 months), and 5 patients are alive with progressive disease with a follow-up between 6 and 8 months; 1 patient is alive with no evidence of disease at 104 months. In four cases, the diagnosis was made recently, with short follow ups of 3 and 4 months. Foci of undifferentiated carcinoma may be confused with solid endometrioid adenocarcinoma erroneously leading to the diagnosis of a grade 3 or a significantly less aggressive grade 2 endometrioid carcinoma. The recognition of undifferentiated carcinoma in an otherwise low-grade endometrioid adenocarcinoma is extremely important because it indicates aggressive behavior. In asynchronous cases, being aware of this association can explain the absence of a second primary. PMID- 16306786 TI - Assessment of uterine wall thickness and position of the vascular plexus in the deep myometrium: implications for the measurement of depth of myometrial invasion of endometrial carcinomas. AB - Accurate discrimination between FIGO stages IB and IC endometrial carcinomas has important prognostic and therapeutic implications, but depth of invasion as a percentage of myometrial thickness can be difficult to ascertain. In such cases, pathologists often presume that infiltration that reaches the arcuate vascular plexus (AVP) in the myometrium indicates >50% myoinvasion. The further assumption is sometimes made that the anterior and posterior uterine walls are of the same thickness. To our knowledge, neither supposition is based on published data. We performed a prospective study of myometrial thickness and the position of the AVP in 50 normal uteruses from patients aged 27 to 84 years. Myometrial thickness varied inversely with age (p < 0.0001); however, anterior and posterior wall myometrial thickness did not differ significantly in the cohort as a whole (p = 0.059) and in individual cases was highly correlated (p < 0.0001). The position of the AVP was variable. On average, its inner limit was situated at a depth of 47.3% of the thickness of the myometrium in both uterine walls, but the position varied between individuals and sometimes differed considerably between the anterior and posterior walls of the same uterus. The position of the AVP did not differ significantly with age. We conclude that carcinomatous infiltration well into or through the AVP usually signifies >50% myoinvasion; however, if infiltration barely extends into the AVP, the depth of invasion should be calculated with reference to the thickness of the myometrium in the opposite uterine wall. PMID- 16306787 TI - Expression of the high-mobility group protein HMGI(Y) in gestational trophoblastic diseases. AB - The high-mobility group protein HMGI(Y) is a member of a family of nonhistone chromosomal proteins, which have been implicated in the regulation of inducible gene transcription, integration of retroviruses into chromosomes, and induction of neoplastic transformation and metastatic progression in cancer cells. The human trophoblast is a tissue that shares proliferation capacity and invasiveness with neoplastic tissues, but in which these processes are tightly regulated. Recently we could show that HMGI(Y) is expressed in the normal human placenta, where it is localized in the nuclei of villous cytotrophoblast, in the anchoring villi at the implantation site and in extravillous (intermediate) trophoblast invading the maternal decidua. In contrast, the majority of the nuclei of the villous syncytiotrophoblast, a terminally differentiated tissue, was negative. The purpose of this study was to investigate the expression pattern of HMGI(Y) in gestational trophoblastic diseases (GTD), which has not been studied so far. To analyze the expression of HMGI(Y), we performed immunohistochemistry on a total of 29 cases of GTD, including 21 hydatidiform moles and 8 choriocarcinomas. Hydatidiform moles showed a positivity for HMGI(Y) in villous cytotrophoblast and in areas of the trophoblast proliferations on the villous surface; villous syncytiotrophoblast was negative. The choriocarcinomas showed strong immunoreactivity in all cases. The expression pattern of HMGI(Y) in gestational trophoblastic diseases indicates that it might play a role in the pathogenesis of GTD and might be potentially useful as an additional diagnostic marker for such lesions. PMID- 16306788 TI - Immunohistochemical evaluation of KIT expression in sarcomas of the gynecologic region. AB - KIT is expressed in most gastrointestinal stromal tumors, and they usually show c kit aberrations (most frequently deletions or deletions coexisting with a single or multiple point mutations). Recently, several studies regarding KIT expression in gynecologic tumors have been reported; however, their outcomes were not consistent. In this study, we immunohistochemically examined KIT expression in sarcomas of the female genital tract and studied the existence of c-kit aberrations to elucidate the characteristics of KIT-positive tumors in the gynecologic region. Formalin-fixed, paraffin-embedded tissues from 25 surgically resected and 1 biopsy specimen from 26 patients were used. Histological diagnoses included 14 uterine leiomyosarcomas, 6 carcinosarcomas, 5 endometrial stromal sarcomas, and 1 vaginal epithelioid sarcoma. Immunohistochemical studies were performed using anti-KIT polyclonal antibody. Only four of the above tumors (15%) were positive for KIT, all of which were carcinosarcomas. Specific KIT immunoreactivity was observed in the only carcinomatous components in one case, in the only sarcomatous component in two cases, and in the both components in one case. However, none of the cases showed c-kit aberrations in exons 9, 11, 13, and 17. Judicious decision is mandatory before applying Imatinib therapy to KIT positive gynecologic tumors. PMID- 16306789 TI - Endometrial mucinous microglandular adenocarcinoma: morphologic, immunohistochemical features, and emphasis in the human papillomavirus status. AB - We report two cases of endometrial microglandular adenocarcinoma, a rare neoplasm, which, in its morphologic features, mimics cervical microglandular hyperplasia and mucinous proliferations of endometrium. The criteria for a correct pathological diagnosis, such as clinical, morphologic, and immunohistochemical data, are emphasized. For the first time, we probed to establish whether endometrial mucinous microglandular adenocarcinoma could be correlated to human papilloma virus (HPV) infection by using polymerase chain reaction amplification (PCR) of tumoral DNA. Similar to previous studies reported in the literature, the present lesions, occurring in postmenopausal women, immunohistochemically showed positivity for B72.3, Ca 125, CEA, Vimentin, estrogen and progesterone receptors, and negativity for p53. Molecular study by PCR amplification of tumor DNA showed no signal for HPV DNA in any of these cases; thus, this variant of endometrial carcinoma is not caused by the HPV infection, but probably by other pathogenetic mechanisms, such as an accumulation of the mutations, which arrive in old age or as the consequence of a peculiar hormonal situation. PMID- 16306790 TI - Ovarian atypical proliferative (borderline) mucinous tumors: gastrointestinal and seromucinous (endocervical-like) types are immunophenotypically distinctive. AB - Ovarian atypical proliferative (borderline) mucinous tumors of gastrointestinal and seromucinous types are considered subtypes within the mucinous tumor category despite the presence of distinctive clinicopathologic features that seromucinous tumors share with pure serous tumors. Immunophenotypic differences have not been extensively investigated. Immunohistochemical studies were performed to compare the expression patterns of cytokeratins 7 and 20 (CK7, CK 20), estrogen and progesterone receptors (ER, PR), CA-125, mesothelin, and WT-1 in 28 tumors of gastrointestinal type and 12 tumors of seromucinous type. Both gastrointestinal and seromucinous type tumors had a high frequency of CK7 expression (93% and 100%, respectively). The gastrointestinal type tumors were characterized by frequent expression of CK20 (86%) and CDX2 (39%), infrequent expression of CA-125 (11%) and mesothelin (7%), and lack of expression of ER, PR, and WT-1. In contrast, the seromucinous type tumors were characterized by frequent expression of ER (100%), PR (67%), CA-125 (92%), and mesothelin (83%), infrequent expression of WT-1 (8%), and lack of expression of CK20 and CDX2. The gastrointestinal and seromucinous types of atypical proliferative mucinous tumors are immunophenotypically distinctive tumors. The former are characterized by expression of markers of gastrointestinal-type differentiation (CK20 and CDX2), whereas the latter are characterized by expression of "mullerian-type" markers (ER, PR, CA-125, and mesothelin). Expression of the latter markers in the seromucinous tumors, which also are expressed in pure serous tumors, and lack of expression of gastrointestinal-type markers, combined with the clinicopathologic features these tumors share with pure serous tumors, support the concept that this subtype is more closely related to serous than gastrointestinal type mucinous tumors and justify the designation "seromucinous." PMID- 16306791 TI - Ovarian biopsy in infertile patients with ovarian dysfunction. AB - Evaluation of ovarian histology is feasible by obtaining ovarian tissue through laparoscopy; however, the role of an ovarian biopsy in an infertility workup is not widely accepted. To gain insight into the role of ovarian biopsy in reproductive medicine, we conducted a cross-sectional study in infertile patients with ovarian dysfunction and correlated follicle stimulating hormone (FSH) and estradiol serum concentrations with ovarian follicular counts. Fifty women were recruited and classified into four groups: premature ovarian failure (POF), chronic anovulation, diminished ovarian reserve, and ovulatory patients (control group). Ovarian endocrine function was assessed by the determination of FSH, luteinizing hormone, and estradiol serum concentrations and correlated with the number of follicles present in the ovarian biopsies obtained by laparoscopy. The number of ovarian follicles observed for each individual biopsy varied extensively. Patients with POF presented significantly lower counts of primordial, primary, and secondary follicles. No significant differences were found in the other groups. In the total sample, primordial follicle counts correlated inversely with serum FSH levels (r = -0.4, p = 0.003) and directly with serum estradiol levels (r = 0.5, p = 0.001) however, such associations no longer remained after adjusting by group. We conclude that ovarian biopsies do not provide additional information to the clinical-hormonal criteria previously established in the workup of infertile patients. Therefore, its use cannot be generalized in the study of infertile patients with ovarian dysfunction. In contrast, ovarian biopsies may be useful to identify patients with POF when the ovarian reserve is likely altered. PMID- 16306792 TI - Immunohistochemical characteristics of mature ovarian cystic teratomas in patients with postoperative recurrence. AB - Because a mature teratoma could recur, we studied the clinical, pathological, and immunohistochemical characteristics of mature ovarian cystic teratomas of 121 patients who underwent surgeries between 1994 and 2002. Recurrence was defined as cases that required secondary surgeries for mature ovarian cystic teratomas. A total of 10 patients were grouped as recurrence, and 111 patients were grouped as non-recurrence. The recurrence group was generally younger (mean, 22.9 +/- 1.26 vs. 32.8 +/- 1.15 years; p < 0.05), had higher tridermal components, and had greater central nervous system (CNS) component expression rate (9 vs. 48%, p < 0.05) compared with the non-recurrence group. The time between primary and secondary surgery averaged 9 years. Our result showed that maturation of teratomas reduced the number of neurons and increased glia. The process is similar to the aging process of brains. The presence of tridermal components and a high synaptophysin/glial fibrillary acidic protein (SP/GFAP) ratio suggested that recurrent ovarian teratomas were capable of both neuronal and glial differentiations. Nestin was expressed in the astrocyte from both the recurrence and non-recurrence groups. Thus, we concluded that nestin was not a useful marker for predicting recurrence and that attention should be given when a high ratio of SP/GFAP is found in lesions. PMID- 16306793 TI - Nicorandil inhibits serum starvation-induced apoptosis in vascular endothelial cells. AB - The Impact Of Nicorandil in Angina (IONA) randomized trial showed a significant reduction in coronary events, in patients with stable angina treated with a KATP channel opener, nicorandil. However, the impact of nicorandil on endothelial apoptosis remains to be examined. We tested the hypothesis that nicorandil has anti-apoptotic effects in endothelial cells (ECs). Apoptosis was induced by serum starvation in the culture media in human umbilical vein endothelial cells. We examined the effects of nicorandil on endothelial cell apoptosis. Cell viability after serum starvation was significantly higher in the nicorandil-treated group compared with the control group (81 +/- 8% vs. 63 +/- 3%, P < 0.01). Apoptosis, as detected by caspase 3 activation and Hoechst 33258 assay, induced by serum starvation was also effectively abrogated by the treatment of nicorandil (100 muM). The protective effects of nicorandil on endothelial survival were significantly inhibited by a specific mitochondrial KATP channel blocker, 5 Hydroxydecanoic acid. A mitochondrial permeability transition pore activator significantly abolished the anti-apoptotic effect of nicorandil in endothelial cells, indicating that the mechanism of protective effect of nicorandil is involved in the mitochondrial apoptotic pathway although it affects neither Bcl-2 nor Bax protein expression levels. In conclusion, nicorandil inhibits serum starvation-induced endothelial cell apoptosis possibly through mitochondrial KATP channels. PMID- 16306794 TI - Pharmacological efficacy of CPU 86017 on hypoxic pulmonary hypertension in rats: mediated by direct inhibition of calcium channels and antioxidant action, but indirect effects on the ET-1 pathway. AB - Endothelin-1 (ET-1) plays a key role in the pathogenesis of pulmonary hypertension. The present study was conducted to examine the effects of a novel compound p-chlorobenzyltetrahydroberberine (CPU 86017) on endothelin-1 system of hypoxia-induced pulmonary hypertension in rats. SD male rats were divided into control, untreated pulmonary hypertension, nifedipine (10 mg/kg p.o.), and CPU 86017 (80, 40, and 20 mg/kg p.o.) groups. The pulmonary hypertension was established by housing the rats in a hypoxic (10 +/- 0.5% oxygen) chamber 8 hours per day for 4 weeks. Hemodynamic and morphologic assessment exhibited a significant increase in the central vein pressure (CVP), right ventricular systolic pressure (RVSP), and pulmonary arteriole remodeling in the pulmonary hypertensive rats, which were improved by CPU 86017 80 and 40 mg/kg administration (P < 0.01). The elevated pulmonary endothelin-1 level and the over active preproET-1 and iNOS mRNA expression were also decreased significantly (P < 0.01) in CPU 86017 groups. The maladjustment of redox enzyme system in pulmonary hypertension rats was corrected after treatment. We concluded that CPU 86017 improves pulmonary hypertension mainly to suppress the endothelin-1 pathway at the upstream and downstream via calcium antagonism and antioxidative action, then, resulting in a relief in pathogenesis of the disease. PMID- 16306795 TI - Effects of angiotensin II receptor blockade with valsartan on pro-inflammatory cytokines in patients with essential hypertension. AB - Chronic inflammation is common in hypertension and acts as an independent determinant of arterial blood pressure. Hypertensive patients are reported to have high circulating levels of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and C-reactive protein (CRP). Recently, angiotensin II receptor blockers (ARBs) have been shown to possess benefits in addition to their ability to lower blood pressure, including anti inflammatory and antioxidative properties within the vasculature. We evaluated the effects of the angiotensin II receptor blocker, valsartan, on these inflammatory cytokines. Thirty-nine patients with essential hypertension participated. These subjects received valsartan, 40 to 80 mg/day. Serum TNF alpha, IL-6, CRP, and serum amyloid A (SAA) were measured before and after 3 months of treatment with valsartan. Valsartan significantly decreased systolic and diastolic blood pressure (160 +/- 16/92 +/- 11 mm Hg to 147 +/- 21/84 +/- 11 mm Hg, P = 0.001/P = 0.001, respectively). Serum TNF-alpha (9.1 +/- 8.6 pg/mL to 6.1 +/- 1.0 pg/mL, P = 0.006) and IL-6 (9.3 +/- 1.7 pg/mL to 8.9 +/- 1.4 pg/mL, P = 0.005) were significantly reduced after treatment with valsartan. However, C reactive protein and serum amyloid A did not change. The angiotensin II receptor blocker, valsartan, may inhibit the development of atherosclerosis by lowering serum pro-inflammatory cytokines. PMID- 16306796 TI - Blood pressure and proteinuria after cessation of a brief renin-angiotensin system blockade in young and adult Lyon hypertensive rats. AB - The aims of the present work were to determine whether a brief renin-angiotensin system blockade with AT1 receptor antagonist or angiotensin-converting enzyme inhibitor may provide long-lasting protection against hypertension and proteinuria in both young and adult Lyon hypertensive (LH) rats. Young pre hypertensive Lyon hypertensive rats were orally treated with 5, 10, or 20 mg/kg/d of losartan, or with 0.4 mg/kg/d of perindopril from 3 to 12 weeks of age. Adult Lyon hypertensive rats were treated with 10 mg/kg/d of losartan, 0.4 mg/kg/d of perindopril, or a combination of both from 12 to 20 weeks of age. Telemetric blood pressure (BP) and urinary protein excretion were evaluated during and after treatment cessation. In young prehypertensive Lyon hypertensive rats, losartan fully prevented the hypertension and proteinuria; these effects were dose dependent and persisted long after treatment withdrawal. When renin-angiotensin system blockade was initiated in adult Lyon hypertensive rats with established hypertension, 10 mg/kg/d of losartan or 0.4 mg/kg/d of perindopril induced a significant regression in both blood pressure (15%-20%) and proteinuria (40%-50%) as did in young Lyon hypertensive rats; the combination treatment produced an additional effect only on blood pressure. After treatment cessation, a reduction in blood pressure persisted in all the pretreated adult Lyon hypertensive rats whereas the effect on proteinuria was less marked. In conclusion, whatever the blocker used, an early renin-angiotensin system blockade in prehypertensive Lyon hypertensive rats induces a durable prevention of hypertension and associated renal alterations. The similar renin-angiotensin system blockade in adult Lyon hypertensive rats provides a regression of hypertension and proteinuria, but the persistence of these beneficial effects is less pronounced than in young Lyon hypertensive rats, thus suggesting that the treatment with renin-angiotensin system blockers should be initiated as early as possible before the full expression of hypertension, to achieve the maximal long-lasting effects in mature stage. PMID- 16306797 TI - Insulin-sensitizing and cardiovascular effects of the sodium-hydrogen exchange inhibitor, cariporide, in the JCR: LA-cp rat and db/db mouse. AB - The effects of the sodium-hydrogen (Na/H) exchange inhibitor cariporide (HOE642), on insulin sensitivity and vascular function were studied in the JCR:LA-cp rat and the db/db mouse. In the insulin-resistant rat, cariporide reduced fasting insulin levels (42%, P < 0.02) and insulin response in a meal tolerance test (50%, P < 0.01), indicating increased insulin sensitivity. The ACE inhibitor, ramipril, used as a reference agent, reduced the insulin response to the meal, but not fasting levels. The EC50 for acetylcholine-mediated relaxation of phenylephrine-precontracted aortic rings was significantly lower in cariporide treated rats (P < 0.002), but not in ramipril-treated rats. Flow response of the coronary circulation to bradykinin was significantly greater in both cariporide- and ramipril-treated rats, (3-fold decrease in the EC50, P < 0.05). Cariporide treated hearts were smaller, slower beating, with greater developed LVP. In the obese db/db mouse, chronic treatment with cariporide obviated vascular hypercontractility and improved endothelial function. Thus, cariporide had beneficial effects on the abnormal insulin metabolism and associated vascular dysfunction in the JCR:LA-cp insulin-resistant rat, which develops advanced cardiovascular disease and ischemic myocardial lesions. It also improved vascular function in a similar mouse model of insulin resistance. These effects were markedly greater than those of ramipril. PMID- 16306798 TI - Mu-opioid receptor blockade protects against circulatory shock and cerebral ischemia during heatstroke. AB - Naltrexone, a nonselective antagonist of opioid receptors, is found to be beneficial in protecting against heatstroke. Further investigation using selective mu, delta, and kappa opioid receptor antagonists are needed to prove the involvement of specific receptors in heatstroke. Rats under sodium pentobarbital anesthesia were exposed to high ambient temperature of 43 degrees C to induce heatstroke. Control rats were exposed to 24 degrees C. In rats treated with normal saline 20 minutes before heat stress, the values for survival time were found to be 89-101 minutes. Intravenous administration of CTAP (a selective mu-opioid receptor antagonist; 50-200 microg/kg), but not nor-binaltorphimine (20 200 microg/kg; a kappa-opioid receptor antagonist) or ICI-174864 (50-500 microg/kg; a delta-opioid receptor antagonist), significantly increased the survival time to new values of 180-212 minutes. In vehicle-treated rats after heatstroke onset, the values for core temperature, intracranial pressure, and the extracellular markers for ischemia (eg, glutamate and lactate/pyruvate ratio) or damage (eg, glycerol) and neuronal damage scores in striatum were significantly higher than those of normothermic controls. In contrast, the values for mean arterial pressure, cerebral perfusion pressure, cerebral blood flow, and brain partial pressure of O2 were significantly lower than those of normothermic controls. The heatstroke-induced hyperthermia, arterial hypotension, intracranial hypertension, cerebral hypoperfusion and hypoxia, and increased levels of cellular ischemia and damage markers in striatum were all significantly attenuated by prior administration of CTAP. The data indicate that prior antagonism of mu-opioid receptors protects against circulatory shock and cerebral ischemia during heatstroke. PMID- 16306799 TI - Endothelium-dependent contractions occur in the aorta of wild-type and COX2-/- knockout but not COX1-/- knockout mice. AB - The present experiments were designed to determine whether or not endothelium dependent contractions can be evoked in the aorta of the mouse, and if so, whether or not deleting the COX1 gene affects the response. Sex differences in the response were also examined. Rings of murine aorta were suspended in a Halpern-Mulvany myograph for recording of isometric force. In the aorta of the male wild type C57BL/b6 mice (36-40 weeks old), both acetylcholine and the calcium ionophore caused endothelium-dependent increases in force in the presence of L-NAME, and these were inhibited by valeryl salicylate (a selective COX1 inhibitor) and S18886 (a selective antagonist of TP receptors). Such endothelium dependent contraction was absent in the aorta of COX1 knockout mice and present in that of COX2 knockout mice. Similar results were obtained in aortas of female wild-type, COX2 and COX1 knockout mice. These experiments reveal the existence of EDCF-mediated contractions in arteries of the mouse. These contractions, as in the aorta of the spontaneously hypertensive rat, are caused by endogenous agonists(s) of TP receptors produced by cyclooxygenase 1, because they are observed in the aortas of COX2 knockout mice but not in aortas of COX1 knockout mice. The present study provides direct evidence that COX1 is indeed the isoform of cyclooxygenase responsible for the production of EDCF. PMID- 16306800 TI - Improvement of EDHF by chronic ACE inhibition declines rapidly after withdrawal in rats with myocardial infarction. AB - Heart failure after myocardial infarction (MI) is associated with endothelial dysfunction. There is conflicting evidence on the exact nature of this endothelial dysfunction and how endothelium-dependent vasodilation is affected by angiotensin-converting enzyme inhibitor (ACE-I) therapy. Furthermore, consequences of acute ACE-I withdrawal are largely unknown. Therefore, we studied the contribution of nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF) to the effects of ACE-I therapy and its withdrawal on endothelial function in MI rats. Rats were subjected to coronary ligation to induce MI and were assigned to quinapril or vehicle from 2 weeks to 8 months post-MI. In parallel, MI rats treated for 14 months with quinapril were subjected to treatment withdrawal for 0, 4, and 6 weeks. Acetylcholine (ACh)-induced relaxation and underlying endothelium-derived mediators were studied in isolated aortic rings. Long-term quinapril (8 months) resulted in markedly improved endothelium-dependent vasodilation in rats with myocardial infarction, which could be attributed to marked improvement in non-NO/prostanoid-mediated relaxation (ie, EDHF). After 14 months of follow-up, maximum vasodilation was still preserved by quinapril. Withdrawal after 14 months of treatment caused significantly impaired ACh-induced EDHF-mediated relaxation within 4 weeks. A marked reduction in EDHF-mediated relaxation caused this impairment. NO-mediated relaxation was unaffected. These findings highlight the importance of EDHF impairment in development of endothelial dysfunction after myocardial infarction and the possibility of improving EDHF-mediated vasodilation with chronic ACE inhibitor therapy. In addition, withdrawal of chronic ACE inhibition after MI should be considered carefully, as profound endothelial dysfunction may develop rapidly. PMID- 16306801 TI - Short-term pioglitazone treatment improves vascular function irrespective of metabolic changes in patients with type 2 diabetes. AB - To determine whether pioglitazone influences endothelial function directly, we examined in a randomized, crossover, placebo-controlled, double-blind trial the effects of 4 weeks of pioglitazone treatment in 20 male type 2 diabetic patients. We conclude that short-term pioglitazone treatment ameliorates endothelial dysfunction in conduit arteries irrespective of significant beneficial changes in plasma levels of insulin, FFA, adiponectin, or CRP in type 2 patients with diabetes. Pioglitazone, a PPARgamma agonist, not only improves insulin resistance and glycemic control but may also have additional beneficial vascular effects in patients with type 2 diabetes. Low-grade inflammation, free fatty acids, and adiponectin may play a role in modulation of vascular function. We studied the effect of 4 weeks of pioglitazone treatment on endothelial function, metabolic changes, and C-reactive protein in patients with type 2 diabetes. A randomized, crossover, placebo-controlled, double-blind trial was performed in which pioglitazone 30 mg once daily was administered to 20 patients with type 2 diabetes on oral antihyperglycemic agents for 4 weeks. Shear stress-induced flow mediated dilation (FMD) of the brachial artery was used as outcome parameter for vascular function. Brachial artery endothelial function was significantly increased by pioglitazone treatment compared with placebo (FMD 5.4 +/- 0.5% versus 3.1 +/- 0.5%, P = 0.001). Endothelium-independent vasodilation was not different between the 2 study periods. Pioglitazone treatment reduced insulin, FFA, and C-reactive protein concentrations compared with placebo (18.3 +/- 2.4 versus 14.8 +/- 2.1 mU/L, P = 0.03; 641 +/- 46 versus 542 +/- 33 mumol/L, P = 0.04; and 3.5 +/- 0.6 mg/L versus 2.6 +/- 0.5 mg/L, P = 0.01; respectively). A significant increase in plasma adiponectin concentration (3.95 +/- 0.57 microg/mL versus 7.59 +/- 0.95 microg/mL, P = 0.002) was also observed. No correlations were found between these metabolic changes and the improvement of conduit artery endothelial function. Short-term pioglitazone treatment ameliorates endothelial dysfunction in conduit arteries irrespective of changes in insulin, FFA, adiponectin, or CRP in type 2 patients with diabetes. PMID- 16306802 TI - Effect of paclitaxel and mesenchymal stem cells seeding on ex vivo vascular endothelial repair and smooth muscle cells growth. AB - Late thrombosis and neointima proliferation after paclitaxel-eluting stents implanting may be related to delayed endothelial cells (ECs) regeneration. This study was to investigate whether mesenchymal stem cells (MSCs) seeding can accelerate endothelial repair and attenuate late smooth muscle cells (SMCs) proliferation after paclitaxel intervention. An ex vivo model of endothelium repair was developed in which rabbit smooth muscle cells were inoculated in the upper chamber and rabbit endothelial cells/human mesenchymal stem cells in the lower chamber of a co-culture system. Paclitaxel (10 nmol/L, 20 min) inhibited smooth muscle cell growth of the confluent endothelial cell group during the observed period. However, increased smooth muscle cells growth was observed in the proliferative endothelial cells group 10 days after paclitaxel intervention. Mesenchymal stem cell seeding inhibited late smooth muscle cell growth incompatible with the effect of proliferative endothelial cells. However, no inhibition on smooth muscle cell growth was observed with mesenchymal stem cell seeding in comparison to the effect of confluent endothelial cells. No vWF but Flk-1 protein was observed in the 25.71% of mesenchymal stem cells after having been co-cultured with rabbit endothelial cells for 5 days. These results indicate that late smooth muscle cell proliferation is closely related to the delayed endothelial cells regeneration after paclitaxel application. Mesenchymal stem cell seeding partly attenuates the late smooth muscle cell proliferation. Mesenchymal stem cells co-cultured with mature endothelial cells have the ability to differentiate toward endothelial cells. PMID- 16306803 TI - Divergent action of calcium channel blockers on ATP-binding cassette protein expression. AB - Calcium channel blockers (CCBs) are widely used in clinical practice, and have been reported to be effective in preventing the progression of atherosclerosis. We examined whether various types of calcium channel blockers affected the expression of ATP binding cassette transporter A1 (ABCA1), a factor contributing to anti-atherogenesis. Undifferentiated monocytic cell line, THP-1 cells were maintained in RPMI 1640 medium and treated with different kinds of calcium channel blockers. Among the calcium channel blockers tested, aranidipine and efonidipine increased ABCA1 protein expression without an increase in ABCA1 mRNA expression, whereas other calcium channel blockers (eg, nifedipine, amlodipine, and nicardipine) or T-type calcium channel blockers (eg, mibefradil and nickel chloride) failed to upregulate ABCA1 expression. H89, a protein kinase A inhibitor inhibited the aranidipine-induced ABCA1 protein expression, whereas genistein (a tyrosine kinase inhibitor), or AG490 (a JAK-2 inhibitor) had no effects. Neither of these inhibitors suppressed the efonidipine-induced ABCA1 protein expression. Intracellular cAMP levels were elevated only by aranidipine, but not by efonidipine. In conclusion, aranidipine and efonidipine have the ability to induce ABCA1 protein by distinct mechanisms; protein kinase A is involved in the aranidipine-induced ABCA1 upregulation. This non-class effect of calcium channel blockers may potentially offer beneficial action in the treatment of hypertensive subjects with atherosclerosis. PMID- 16306804 TI - Cardioprotection with adenosine A2 receptor activation at reperfusion. AB - Pre-ischemic treatment is seldom possible in the clinical setting of acute myocardial infarction. Thus, to successfully save myocardium from infarction, it is required that protective interventions must be effective when applied after ischemia has begun or at the onset of reperfusion. Unfortunately, in spite of a large body of experimental data showing that various interventions are cardioprotective at reperfusion, no specific therapy has yet been established to be clinically applicable. However, recent data from several laboratories have shown that adenosine and its analogues given at reperfusion can markedly protect the heart from ischemia/reperfusion injury. While the experimental data suggest that factors such as adenosine A2 receptor activation, anti-neutrophil effect, attenuation of free radical generation, increased nitric oxide (NO) availability, activation of the PI3-kinase/Akt pathway and ERK, prevention of mitochondrial damage, and anti-apoptotic effects may be involved in the protective effect of adenosine or its analogues, the exact receptor subtype(s), the detailed signaling mechanisms, and interaction between those individual factors are still unknown. A definite answer to these unsolved problems will offer insights into the mechanisms of cardioprotection at reperfusion, and will be critical for developing a successful therapeutic strategy to salvage ischemic myocardium in patients with acute myocardial infarction. PMID- 16306805 TI - Captopril restores endothelium-dependent relaxation induced by advanced oxidation protein products in rat aorta. AB - To explore whether advanced oxidation protein products (AOPP) can cause endothelial dysfunction in vitro, and whether captopril exerts beneficial effect on impaired endothelium-dependent relaxation induced by exogenous advanced oxidation protein products and to investigate the potential mechanisms. Both the Acetylcholine (ACh)-induced endothelium-dependent relaxation (EDR), sodium nitroprusside-induced endothelium-independent relaxation of aortic rings were measured by recording isometric tension after the rings were exposed to AOPP-BSA in the absence or presence of captopril to assess the injury effect of AOPP-BSA and the protective effect of captopril on the aortic endothelium, respectively. Co-incubation of aortic rings with AOPP-BSA (3 mmol/L) for 90 minutes resulted in a significant inhibition of EDR to ACh, but had no effects on endothelium independent relaxation to SNP. After incubation of the rings in the co-presence of captopril (3 to 30 micromol/L) or enalaprilat (30 micromol/L) with AOPP-BSA (3 mmol/L) for 90 minutes, captopril significantly and enalaprilat only partly attenuated the inhibition of EDR induced by AOPP-BSA. This protective effect of captopril (30 micromol/L) was abolished by N-nitro-L-arginine methyl ester (10 micromol/L), an inhibitor of nitric oxide synthase. Furthermore, the superoxide anion scavenger superoxide dismutase (SOD, 200 U/mL), and the nitric oxide precursor L-arginine (3 mmol/L) also ameliorated the impaired EDR caused by AOPP BSA. But D-arginine had no effect on the impaired EDR caused by AOPP-BSA. AOPP BSA can trigger endothelial dysfunction and captopril can protect the endothelium against functional damage induced by AOPP-BSA in rat aorta, increase nitric oxide bioavailability. The mechanisms of endothelial dysfunction induced by AOPP-BSA may include the decrease of NO and the generation of oxygen-free radicals. PMID- 16306806 TI - Flow-independent myocardial ischemia induced by endothelin-1: an NADH fluorescence analysis. AB - The endothelin-1 (ET-1) is known to cause myocardial ischemia; however, whether this effect is entirely dependent on vasoconstriction is uncertain. The aim of this study was to characterize the myocardial ischemia after the intracoronary administration of endothelin-1, and compare it with that induced by coronary stenosis. In the left anterior descending coronary artery of 15 dogs, a mild inflow reduction (30%) was produced for 10 minutes using intracoronary endothelin 1 (46 +/- 33 pmol/min) or coronary stenosis. The hearts were rapidly cross sectioned at short axial plane and freeze-clamped within 120 milliseconds using a specially developed device to visualize and quantify the area of ischemia (%IA) with NADH fluorescence photography. The %IA was larger in the endothelin-1 group than in the stenosis group (66 +/- 23 versus 18 +/- 18, P = 0.0005); furthermore, the ischemia was transmural in the ET-1 group, but limited to subendocardium in the stenosis group. ET-1 increased the coronary arterial resistance especially in subepicardial region and produced smaller ischemic foci in microcirculation. The mechanism of larger ischemia produced by ET-1 might depend on pro-ischemic effects on myocytes and vasoconstriction of the coronary microcirculation, predominantly in the subepicardium in vivo. PMID- 16306807 TI - Pioglitazone mimics preconditioning in the isolated perfused rat heart: a role for the prosurvival kinases PI3K and P42/44MAPK. AB - Ischemic preconditioning, the most powerful protection against infarction, activates PI3Kinase (PI3K)/AKT and P42/44MAPK. Pioglitazone, a thiazolidinedione and PPARgamma receptor agonist used in Type II diabetes treatment, has been shown to activate these kinase cascades. We therefore hypothesized that pioglitazone could protect the myocardium when given prior to myocardial ischemia/reperfusion injury. Langendorff perfused rat hearts underwent 40 minutes of stabilization then 35 minutes of regional ischemia and 120 minutes of reperfusion (control) or Pioglitazone (1, 2, 5, and 10 microM)-given before ischemia. Additional groups underwent the same protocol but with either PI3K inhibitors (15 microM LY294002 or 100 nM wortmannin) or P42/44MAPK inhibitors (10 microM U0126 or 10 microM PD98059) given either during stabilization or at reperfusion. Infarct size was determined as a percentage of risk zone (I/R%). Pioglitazone (2 microM) significantly reduced I/R% compared with control (25.4 +/- 3.1 versus 47.3 +/- 3.4; P < 0.05). This protection was abolished by PI3K inhibitors (pioglitazone+LY294002 46.5 +/- 5.0, pioglitazone + wortmannin 48.8 +/- 4.6 versus pioglitazone alone 25.4 +/- 3.1; P < or = 0.05) but not by P42/44MAPK inhibitors (pioglitazone+U0126 30.7 +/- 5.7, pioglitazone + PD98059 28.5 +/- 6.3 versus pioglitazone alone 25.4 +/- 3.1; P < or = 0.05) given in stabilization. However when the inhibitors were given at reperfusion, the protection was abrogated by blocking either pathway (pioglitazone+LY294002 49.8 +/- 3.1, pioglitazone+U0126 48.7 +/- 3.7 versus pioglitazone alone 25.4 +/- 3.1; P < or = 0.05). In conclusion pioglitazone induced significant protection against ischemia/reperfusion injury when administered prior to ischemia. This protection appears to involve PI3K and P42/44MAPK. PMID- 16306808 TI - Blood pressure responses of endothelin-1 1-31 within the rostral ventrolateral medulla through conversion to endothelin-1 1-21. AB - Endothelin-1 1-31 (ET-1 1-31), a novel member of the endothelin family comprising 31 amino acids and derived from the selective hydrolysis of big ET-1 by chymase, directly activates endothelin receptors or converts to ET-1 1-21 by ET converting enzyme (ECE). The cardiovascular effects of central ET-1 1-31 are not identified. The present study was designed to investigate the cardiovascular actions of ET-1 1-31 within the rostral ventrolateral medulla (RVLM) in anesthetized rats. Bilateral injection of ET-1 1-31 (0.5, 1, and 2 pmol for each side) into the rostral ventrolateral medulla produced an initial pressor and/or a long-lasting hypotensive action but did not affect HR. Unilateral microinjection of 2 and 4 pmol of ET-1 1-31 into the rostral ventrolateral medulla only produced a significant (P < 0.05) transient increase in blood pressure by an average of 13 and 12 mm Hg, respectively, whereas unilateral microinjection of 8 pmol of ET-1 1 31 produced a sustained fall in blood pressure (from 92 +/- 6 to 69 +/- 8 mm Hg, P < 0.05). The transient pressor effect of unilaterally injecting ET-1 1-31 (4 pmol) into the rostral ventrolateral medulla was completely abolished by pretreatment with either ETA receptor antagonist BQ123 (83 +/- 2 versus 84 +/- 5 mm Hg, P > 0.05) or ET converting enzyme inhibitor phosphoramidon (99 +/- 5 versus 99 +/- 7 mm Hg, P > 0.05) but not ETB receptor antagonist IRL1038 (89 +/- 6 versus 96 +/- 7 mm Hg, P < 0.05). In addition, prior injection of phosphoramidon also completely abolished the long-lasting hypotension of intra RVLM ET-1 1-31 (8 pmol) but did not modify the depressor action of intra-RVLM ET 1 1-21 (from 100 +/- 6 to 76 +/- 8 mm Hg, P < 0.05). In conclusion, the current results suggest that the cardiovascular effects of intra-RVLM ET-1 1-31 might be the result of conversion of ET-1 1-31 to ET-1 1-21 through activation of ETA receptors. PMID- 16306809 TI - Duration of the beneficial effects of levosimendan in decompensated heart failure as measured by echocardiographic indices and B-type natriuretic peptide. AB - Levosimendan is effective in the treatment of decompensated heart failure. The beneficial effects of a single dose of levosimendan last much longer than those of other inotropes. However, the exact duration of the beneficial effects is unknown. We prospectively determined the duration of the cardiac effects, as measured by echocardiography, of levosimendan (LS) following a 24-hour infusion regimen in patients with decompensated heart failure (DHF). The effects of LS on plasma B-type natriuretic peptide (BNP) were also examined. Twenty patients with DHF displaying (1) deteriorating symptoms despite optimal oral therapy, (2) left ventricular ejection fractions (LVEF) < 35%, and (3) cardiac indices of < 2.5 L/m/min received 24 hours of LS infusion. Echocardiography and BNP measurements were performed pre- and postinfusion and were reassessed on days 7, 30, and 90. Left ventricular systolic function indices (cardiac output and LVEF), LV filling pressure indices, and right ventricular systolic function indices all improved following LS treatment. Most of these improvements were sustained for at least 7 days (P < 0.05) and returned to baseline by day 30 postinfusion and remained so on day 90. Plasma BNP also displayed the same pattern of transient improvements. In conclusion, LS transiently improved the cardiac function, and the effects lasted for at least 7 days after discontinuation of infusion. Most effects, except LVEF, were not significantly different from baseline on day 30. PMID- 16306810 TI - Diminished responses to nifedipine imply severe end-organ damage in spontaneously hypertensive rats. AB - This study was designed to investigate the effect of end-organ damage (EOD), the initial blood pressure levels, and baroreflex sensitivity (BRS) on the blood pressure-lowering effect of nifedipine in spontaneously hypertensive rats (SHRs). Nifedipine was intravenously administered at a dose of 1 mg/kg. BP was continuously recorded in the conscious state before and after nifedipine administration. BRS was determined before drug administration. Two days after the blood pressure (BP) recording, rats were killed for organ-damage evaluation. Univariate correlation analysis showed that BP changes induced by nifedipine injection were negatively correlated with EOD score and aortic weight/length but positively correlated with left kidney weight/body weight and basal BP levels. Stepwise multiple linear regression analysis demonstrated that increase in overall end-organ damage was most significantly related to the decrease in hypotensive effect of nifedipine; increase in aortic hypertrophy was also related to a decreased fall in systolic and diastolic BP induced by nifedipine, whereas increase in initial BP levels was associated with increased hypotensive effect of nifedipine. In conclusion, the severity of overall EOD contributed more than basal BP levels to the diminished responses to nifedipine, and aortic hypertrophy was also involved in diminished drug responses. PMID- 16306811 TI - An epoxide hydrolase inhibitor, 12-(3-adamantan-1-yl-ureido)dodecanoic acid (AUDA), reduces ischemic cerebral infarct size in stroke-prone spontaneously hypertensive rats. AB - Soluble epoxide hydrolase (sEH) inhibitors have been demonstrated to have cardiovascular protective actions. This hydrolase enzyme converts fatty acid epoxides to their corresponding diols, and this conversion can alter the biologic activity of these metabolites. We hypothesized that 12-(3-adamantan-1-yl ureido)dodecanoic acid (AUDA), a sEH inhibitor, would protect stroke-prone spontaneously hypertensive rats from cerebral ischemia. AUDA was administered to 6-week-old male rats for 6 weeks, during which blood pressure was measured by telemetry. Cerebral ischemia was induced by middle cerebral artery occlusion, the size of the cerebral infarct was assessed after 6 hours of ischemia, and the results were expressed as a percentage of the hemisphere infarcted (%HI). Vascular structure and function were assessed using a pressurized arteriograph. Plasma levels of AUDA at the end of the treatment period averaged 5.0 +/- 0.4 ng/mL, and the urinary excretion rate was 99 +/- 21 ng/d. AUDA-treated rats had significantly smaller cerebral infarcts than control rats (36 +/- 4% vs 53 +/- 4% HI, treated versus control, P < 0.05, n = 6). This difference occurred independently of changes in blood pressure. AUDA treatment increased the passive compliance of the cerebral vessels but had no effect on vascular structure. The results of this study provide novel evidence suggesting that the sEH inhibitor AUDA is a possible therapeutic agent for ischemic stroke. PMID- 16306812 TI - Dissociation between metabolic and efficiency effects of perhexiline in normoxic rat myocardium. AB - The antianginal agent perhexiline inhibits rat cardiac carnitine palmitoyltransferase-1 (CPT-1) and CPT-2, key enzymes for mitochondrial transport of long-chain fatty acids. We tested the hypothesis that perhexiline, in therapeutic concentrations (2 microM), inhibits palmitate oxidation and enhances glucose oxidation in isolated rat cardiomyocytes and in the working rat heart, thereby increasing efficiency of oxygen utilization. In isolated cardiomyocytes, perhexiline (2 microM) exerted no acute effects on palmitate oxidation, but after 48 hours pre-exposure oxidation was inhibited by perhexiline (2 to 10 microM) by 15% to 35% (P < 0.0002). In non-ischemic working rat hearts (3%BSA, 0.4 mM palmitate, 11 mM glucose, 100 microU/mL insulin) perhexiline (2 microM) had no significant acute effect on cardiac efficiency, palmitate or glucose oxidation, but 24 hours pretreatment with transdermal perhexiline increased cardiac work (by 29%, P < 0.05) and cardiac efficiency (by 30%, P < 0.02) without significant effects on palmitate oxidation. The selective CPT-1 inhibitor oxfenicine (2 mM) inhibited palmitate oxidation and enhanced glucose oxidation, but failed to enhance cardiac efficiency. In conclusion, in the non-ischemic working rat heart, perhexiline increases myocardial efficiency by a mechanism(s) that is largely or entirely independent of its effects on CPT. Effects on cardiac efficiency during ischemia, and with changes in fatty acid oxidation after longer perhexiline pretreatment remain to be determined. PMID- 16306813 TI - Pomegranate flower extract diminishes cardiac fibrosis in Zucker diabetic fatty rats: modulation of cardiac endothelin-1 and nuclear factor-kappaB pathways. AB - The diabetic heart shows increased fibrosis, which impairs cardiac function. Endothelin (ET)-1 and nuclear factor-kappaB (NF-kappaB) interactively regulate fibroblast growth. We have recently demonstrated that Punica granatum flower (PGF), a Unani anti-diabetic medicine, is a dual activator of peroxisome proliferator-activated receptor (PPAR)-alpha and -gamma, and improves hyperglycemia, hyperlipidemia, and fatty heart in Zucker diabetic fatty (ZDF) rat, a genetic animal model of type 2 diabetes and obesity. Here, we demonstrated that six-week treatment with PGF extract (500 mg/kg, p.o.) in Zucker diabetic fatty rats reduced the ratios of van Gieson-stained interstitial collagen deposit area to total left ventricular area and perivascular collagen deposit areas to coronary artery media area in the heart. This was accompanied by suppression of overexpressed cardiac fibronectin and collagen I and III mRNAs. Punica granatum flower extract reduced the up-regulated cardiac mRNA expression of ET-1, ETA, inhibitor-kappaBbeta and c-jun, and normalized the down-regulated mRNA expression of inhibitor-kappaBalpha in Zucker diabetic fatty rats. In vitro, Punica granatum flower extract and its components oleanolic acid, ursolic acid, and gallic acid inhibited lipopolysaccharide-induced NF-kappaB activation in macrophages. Our findings indicate that Punica granatum flower extract diminishes cardiac fibrosis in Zucker diabetic fatty rats, at least in part, by modulating cardiac ET-1 and NF-kappaB signaling. PMID- 16306814 TI - Pharmacokinetic and pharmacodynamic interaction between irbesartan and hydrochlorothiazide in renal hypertensive dogs. AB - Combined irbesartan/hydrochlorothiazide (HCTZ) formulations are often used clinically. Pharmacokinetic-pharmacodynamic (PK/PD) modeling was applied to investigate the pharmacokinetic and pharmacodynamic interaction between irbesartan and HCTZ in renal hypertensive dogs at non-steady-state and steady state. The renal hypertensive dogs were treated with oral irbesartan alone, or HCTZ alone, or the combination of irbesartan and HCTZ for 8 days. Blood pressure and plasma concentrations were measured and pharmacokinetic-pharmacodynamic parameters were analyzed. Irbesartan showed a two-compartment model pharmacokinetic profile. The concentration-time course of irbesartan was not changed by HCTZ, but irbesartan increased the peak plasma concentration and area under the curve of HCTZ at steady-state. HCTZ had no blood pressure lowering effect at non-steady-state. Irbesartan plus HCTZ had greater blood pressure lowering action than irbesartan alone. HCTZ increased actions of irbesartan. Hysteresis loops were found between effect and plasma concentrations of irbesartan after a single dose. However, hysteresis loops disappeared at steady state with more rapid realization of maximum concentration and effects. The relationship between effects and effect-compartment concentrations of the drugs was represented by a sigmoid Emax model. The results suggest synergistic pharmacodynamic interaction between irbesartan and HCTZ in renal hypertensive dogs and some differences of pharmacokinetic-pharmacodynamic properties between irbesartan and irbesartan/HCTZ combinations at non-steady-state and steady state. PMID- 16306815 TI - The impact of instrument fracture on outcome of endodontic treatment. AB - Adoption of rotary nickel-titanium instruments has renewed concerns regarding instrument fracture and its consequences. The frequency of instrument fracture and its impact on treatment outcome were determined from an analysis of specialist endodontic practice records involving 8460 cases. A case-control study of treatment outcomes was conducted on a subset of 146 teeth with a retained instrument fragment (plus 146 matched controls), for which clinical and radiographic follow-up of at least 1 year was available. Masked radiographs were assessed by two calibrated examiners. Overall prevalence of retained fractured instruments was 3.3% of treated teeth. In the case-control study, overall healing rates were 91.8% for cases with a fractured instrument and 94.5% for matched controls (p > 0.05, Fisher's exact test). Healing in both groups was lower in teeth with a preoperative periapical radiolucency (86.7% versus 92.9%, p > 0.05). In the hands of skilled endodontists prognosis was not significantly affected by the presence of a retained fractured instrument. PMID- 16306816 TI - Enterococcus spp. in endodontically treated teeth with and without periradicular lesions. AB - The purpose of this study was to determine if Enterococcus spp. are more prevalent in endodontically treated teeth with periradicular lesions compared with teeth that require retreatment but have no periradicular rarefaction. Fifty eight teeth that had received root canal therapy more than 1 yr previously and required retreatment were included. Designation of lesion versus no lesion was determined by two experienced endodontists. DNA extraction and PCR amplification were performed using ubiquitous 16S rDNA bacterial primers, as well as Enterococcus spp.-specific primers. The results showed that the overall prevalence of bacteria was 90% and Enterococcus spp. was 12%. chi analysis revealed a statistically significant relationship between the presence of a lesion and the presence of bacteria, as detected by the universal primers (p = 0.032). Using logistic regression, a statistically significant relationship was found between teeth with normal periapex and the presence of Enterococcus spp. (p = 0.023). This study revealed that bacteria are significantly associated with endodontic treatment failure but enterococci are not associated with disease. PMID- 16306817 TI - The effect of cavity preparation on substance P expression in human dental pulp. AB - Substance P (SP) plays an important role during neurogenic inflammation of dental pulp. The purpose of this study was to use a radioimmunoassay for determining the effect of cavity preparation on SP expression in healthy human dental pulp. Ten pulp samples were obtained from healthy premolars where extraction was indicated for orthodontic reasons. Deep cavity preparation (<1 mm remaining dentine thickness) was performed before extraction in five of these bicuspids. All samples were processed and I-SP labeled; SP was quantified by competition assay. The results revealed SP expression in all human pulp samples. Mann-Whitney's U test revealed statistically significant higher expression in pulp from teeth where cavity preparation had been performed compared to control values (p < 0.05). These findings suggest that SP is released during common dental procedures (such as cavity preparation) and its expression may have an important clinical significance in terms of experiencing inflammation and pain. PMID- 16306818 TI - The upregulation of matrix metalloproteinase-9 in inflamed human dental pulps. AB - The matrix metalloproteinases (MMPs) participate in a wide variety of extracellular matrix degradation. Detailed knowledge of MMPs may be important for understanding the pathogenesis of pulpal inflammation. The purpose of this study was to compare MMP-9 expression in clinically healthy human pulp and inflamed human pulp tissue specimens. We compared the levels of MMP-9 between clinically healthy pulp and inflamed pulp tissues by using the semi-quantitative reverse transcriptase polymerase chain reaction analysis. In addition, immunohistochemistry was used to identify the in situ localization of MMP-9 expression in pulp specimens. MMP-9 mRNA gene was found to be increased in inflamed pulps as compared with clinically healthy pulp tissues (p < 0.05). The results from immunohistochemistry demonstrated that MMP-9 expression was significantly higher in the inflamed pulps than clinically healthy pulps (p < 0.05). MMP-9 stain was detected in the odontoblasts, fibroblasts, inflammatory infiltrates, and endothelial cells. Taken together, MMP-9 may play an important role in the pathogenesis of pulpal inflammation. PMID- 16306819 TI - Clinical efficacy of treatment procedures in endodontic infection control and one year follow-up of periapical healing. AB - The objective was to evaluate the clinical efficacy of chemomechanical preparation of the root canals with sodium hypochlorite and interappointment medication with calcium hydroxide in the control of root canal infection and healing of periapical lesions. Fifty teeth diagnosed with chronic apical periodontitis were randomly allocated to one of three treatments: Single visit (SV group, n = 20), calcium hydroxide for one week (CH group n = 18), or leaving the canal empty but sealed for one week (EC group, n = 12). Microbiological samples were taken to monitor the infection during treatment. Periapical healing was controlled radiographically following the change in the periapical index at 52 wk and analyzed using one-way ANOVA. All cases showed microbiological growth in the beginning of the treatment. After mechanical preparation and irrigation with sodium hypochlorite in the first appointment, 20 to 33% of the cases showed growth. At the second appointment 33% of the cases in the CH group revealed bacteria, whereas the EC group showed remarkably more culture positive cases (67%). Sodium hypochlorite was effective also at the second appointment and only two teeth remained culture positive. Only minor differences in periapical healing were observed between the treatment groups. However, bacterial growth at the second appointment had a significant negative impact on healing of the periapical lesion (p < 0.01). The present study indicates good clinical efficacy of sodium hypochlorite irrigation in the control of root canal infection. Calcium hydroxide dressing between the appointments did not show the expected effect in disinfection the root canal system and treatment outcome, indicating the need to develop more efficient inter-appointment dressings. PMID- 16306820 TI - The role of environmental changes on monospecies biofilm formation on root canal wall by Enterococcus faecalis. AB - Biofilm mode of growth is a strategy in microorganisms to survive harsh growth conditions. Although previous studies have established the ability of Enterococcus faecalis to survive postendodontic environmental conditions, the effect of such conditions on the ultrastructural and physiochemical features of E. faecalis biofilm has received less attention. This study aims to evaluate the effect of different growth conditions on the characteristics of E. faecalis biofilm on root canal, and the penetration of E. faecalis into dentinal tubules. Forty-five intact noncarious human maxillary molars were experimented under nutrient-rich, nutrient-deprived, aerobic, and anaerobic conditions for a period of 21 days. Scanning Electron Microscopy with Energy Dispersive X-ray microanalysis, Laser Confocal Scanning Microscopy and Light microscopic examinations were carried out. The microscopic analysis highlighted a distinct variation in the ultrastructure of the biofilms formed under different experimental conditions. The EDX microanalysis showed a significant increase in the levels of Calcium (Ca) in the biofilm structures formed under anaerobic nutrient-deprived condition (p < 0.001). The depth of bacterial penetration was significantly greater in nutrient-rich condition (p < 0.001). This study demonstrated distinct ultrastructural and physiochemical properties of the biofilms formed and dentinal tubular penetration of E. faecalis under different conditions. PMID- 16306821 TI - A comparison of the machining efficiency of two brands of stainless steel endodontic hand files. AB - The purpose of this study was to compare the machining efficiency of a flexible stainless steel K-type hand file recently introduced by Brasseler USA ("F-Style files"), with that of a well-studied Endodontic instrument ("Flex-o-files," Maillefer Dentsply), that has been on the market for more than 20 years. The comparison of machining efficiencies of these two brands of files was conducted on both dentin and plexiglas substrates, using an apparatus that allowed a constant force to be applied from each file being tested. The results indicated that the Brasseler instruments had poorer machining efficiency than those of the Maillefer files. PMID- 16306822 TI - An in vitro model to investigate filling of lateral canals. AB - Aims of this work were to examine lateral canals in extracted teeth, to propose a new technique to produce artificial lateral canals, and to compare two obturation techniques. Cleared roots were examined to record measure and shape of lateral canals. Artificial lateral canals were prepared on human demineralized teeth before final clearing. Specimens were divided in two groups: canals of group 1 were filled with Schilder's technique, canals of group 2 were filled with vertical compaction with apical backfilling. Stereomicroscopic analysis of lateral canal filling revealed lower filling rates in apical canals compared to coronal ones and higher filling rates with "vertical compaction with apical backfilling" compared to Schilder's group. The tested procedure appears to be a reliable technique to obtain standardized lateral canals and to compare filling procedures. PMID- 16306823 TI - Atomic force microscopy study of stainless-steel and nickel-titanium files. AB - Atomic force microscopy (AFM) is a well established and documented tool for materials investigation. The aim of this study was to evaluate the topography of conventional stainless-steel files and both hand and rotary nickel-titanium (NiTi) files by using AFM. One endodontic file of each of the following was selected: stainless-steel K-file Dentsply, stainless-steel K-file Moyco, hand NiTi K-file Nitiflex, hand NiTi Greater Taper, rotary NiTi Greater Taper, and rotary NiTi Quantec. The analyses were performed on twenty different points located along a 3-mm section starting at the tip of each file. Root mean square (RMS) parameters for contact mode imaging microscopy variations were measured. The differences between RMS values were tested by ANOVA with Fisher's protected LSD test for multiple comparisons (p < 0.05). RMS of depth profile data was used to determine any statistically significant difference in vertical amplitude. According to results, all instruments showed topographic irregularities distributed on surface. Endodontic files manufactured by the same method and alloy demonstrated significant differences, whereas no significant differences were found for instruments produced by different alloys and methods (p < 0.05). The hand NiTi Greater Taper, rotary NiTi Greater Taper, and rotary NiTi Quantec showed greater values of vertical amplitude topography compared to K-Dentsply and Nitiflex files (p < 0.05). The AFM technique proved to be a valuable research tool in the investigation of endodontic files topography. PMID- 16306824 TI - Apical leakage of resin based root canal sealers with a new computerized fluid filtration meter. AB - In this in vitro study, the apical leakage of three root-canal sealers: AH Plus, Diaket, and EndoREZ was evaluated using a new computerized fluid filtration meter. Forty-five extracted human premolar teeth with single root and canal were used. The coronal part of each tooth was removed and the root canals were prepared using GT Rotary files and crown-down technique. The roots were randomly divided into three groups of 15 samples, filled with one of the test materials and gutta-percha cones by the cold lateral condensation technique and were stored at 37 degrees C and 100% humidity for 7 days. One-week later, apical parts of roots of 10 +/- 0.05 mm were attached to computerized fluid filtration meter. Apical leakage quantity was determined as microl/cmH(2)O/min(-1). Statistical analysis indicated that root fillings with Diaket in combination with cold lateral condensation technique showed lower apical leakage than the others (p < 0.05). In addition, this new computerized fluid filtration meter allowed quantitative measurement of leakage easily. As it is a newly developed device to measure apical leakage of endodontic sealers, the reliability of it needed to be tested. PMID- 16306826 TI - Fracture resistance of human root dentin exposed to intracanal calcium hydroxide. AB - The purpose of the present study was to determine if exposure to intracanal calcium hydroxide [Ca(OH2)] alters the fracture resistance of human root dentin. One hundred and two freshly extracted single rooted human teeth divided into three groups of 34 teeth each. Coronal access and endodontic instrumentation using round burs, stainless steel files, and Profile GT rotary files were completed for each tooth. The prepared root canal system of each tooth was filled with saline solution (group 1), USP Ca(OH)2 (group 2), or Metapaste (group 3). The apicies and access openings were sealed with composite resin and the teeth were immersed in saline. After 30 days, the roots of 17 teeth from each group were sectioned horizontally into 1-mm thick disks and each disk was loaded to fracture at 2.5 mm/min with a SATEC universal-testing machine. After 180 days the same procedure was performed on the remaining 17 teeth in each of the 3 groups. The peak load at fracture was measured for each dentin disk. Data were analyzed using one-way ANOVA and a post hoc Student-Newman-Keuls test. After 30 days exposure to the test solution, there was no difference in the peak load at fracture for the three groups of teeth. However, after 180 days, the roots of the teeth exposed to USP Ca(OH)2 showed a significant decrease in peak load at fracture when compared to the 30-day groups and the 180-day groups exposed to saline or Metapaste. PMID- 16306825 TI - Does hybridization of intraradicular dentin really improve fiber post retention in endodontically treated teeth? AB - This study tested the hypothesis that hybridization of intraradicular dentin eliminates interfacial gaps, thereby improving the coronal seal and retention of teeth restored with fiber posts. Post spaces were bonded with two types of fiber posts, using the corresponding etch-and-rinse adhesives and dual-cured resin cements. Longitudinal sections of the interfaces were examined for dentin hybridization in the coronal- and middle-thirds of the root canals. Resin replicas of these sections were evaluated for interfacial gap formation. Although intraradicular dentin hybridization was not compromised irrespective of whether the adhesives were light-cured before cementation, the universal occurrence of interfacial gaps along the hybrid layer surface or the post-cement interface reflects the challenge in bonding to post spaces with low compliance and high C factors. The clinical success associated with bonded fiber posts is probably due predominantly to frictional retention. PMID- 16306827 TI - Endodontic treatment of a large cyst-like periradicular lesion using a combination of antibiotic drugs: a case report. AB - This case report describes the endodontic treatment of a large cyst-like periradicular lesion using a combination of antibiotic drugs. A 15-yr-old boy with a large cyst-like lesion from the apices of his teeth 29 to 31. Conservative root canal treatment was performed (tooth #30). During treatment procedure, 2.5% sodium hypochlorite solution was used for irrigation and a combination of antibiotic drugs was used for the intracanal dressing. Periapical healing was observed 3-month after initial treatment and continued at the 7-month review. PMID- 16306832 TI - Selective nerve root injections can predict surgical outcome for lumbar and cervical radiculopathy: comparison to magnetic resonance imaging. AB - OBJECTIVE: Diagnostic selective nerve root injection (SNI) results were analyzed in 101 patients who underwent lumbar or cervical decompression for radiculopathy and compared to surgical outcome 1 year postoperatively. A comparison of surgical outcomes was also examined between magnetic resonance imaging (MRI) and SNI results. RESULTS: Of the 101 patients, 91 (90%) had positive and 10 had negative SNI results at the level operated. Ninety-one percent of the patients with a positive SNI had good surgical outcomes, whereas 60% of the patients with a negative SNI had good outcomes. Of the patients with a positive MRI result, 87% had good surgical outcomes, whereas a similar percentage of the patients with a negative MRI (85%) had good surgical outcomes. When findings between SNI and MRI differed (n = 20), surgery at a level consistent with the SNI was more strongly associated with a good surgical outcome. Of the patients with a poor surgical outcome, surgery was most often performed at a level inconsistent with the SNI finding. CONCLUSIONS: Our study found that a diagnostic SNI can safely and accurately discern the presence or absence of cervical or lumbar radiculopathy. The diagnostic SNI can persuade surgeons from operating on an initially suspicious, but incorrect, level of radiculopathy. In cases where MRI findings are equivocal, multilevel, and/or do not agree with the patient's symptoms, the result of a negative diagnostic SNI (ie, lack of presence of radiculopathy) becomes superior in predicting the absence of an offending lesion. PMID- 16306833 TI - Is high level of disability an indication for spinal fusion? Analysis of long term outcome after posterior lumbar interbody fusion using carbon fiber cages. AB - OBJECTIVE: Posterior lumbar interbody fusion is a recognized procedure for the treatment of back pain associated with degenerative disc disease and segmental instability. It allows decompression of the spinal canal and circumferential fusion through a single posterior incision. METHODS: Sixty-five consecutive patients who underwent posterior lumbar interbody fusion using carbon cages and pedicle fixation between 1993 and 2000 were recruited and contacted with a postal survey. Clinical outcome was assessed by the postoperative clinical findings and complications and the fusion rate, which was assessed using the scoring system described by Brantigan and Steffee. Functional outcome was measured by using improvement in the Oswestry Disability Index, return to work, and satisfaction with the surgical outcome. The determinants of functional relief were analyzed against the improvement in disability using multiple regression analysis. RESULTS: The mean postoperative duration at the time of the study was 4.4 years. Overall radiologic fusion rate was 98%. There was a significant improvement in Oswestry Disability Index (P < 0.01). There was 84% satisfaction with the surgical procedure and 61% return to predisease activity level and full employment. We found preoperative level of disability to be the best determinant of functional recovery irrespective of age or the degree of psychological morbidity and litigation (P < 0.01). CONCLUSION: The combination of posterior lumbar interbody fusion and posterior instrumented fusion is a safe and effective method of achieving circumferential segmental fusion. A direct relationship between preoperative level of disability and functional recovery suggests that disability should be measured preoperatively and spinal fusion should be performed to alleviate disability caused by degenerative spine. PMID- 16306834 TI - Posterior fixation of thoracolumbar burst fracture: short-segment pedicle fixation versus long-segment instrumentation. AB - OBJECTIVE: The treatment of thoracolumbar burst fracture is a controversial issue. Short-segment (SS) pedicle fixation has become a popular treatment option. However, there are several studies regarding the high rate of failure. The aim of this prospective study was to compare SS versus long-segment (LS) instrumentation. METHODS: For this purpose, 18 consecutive patients were assigned to two groups. Group 1 included nine patients treated by SS pedicle fixation, whereas group 2 included nine patients treated by LS instrumentation. SS instrumentation was pedicle fixation one level above and below the fractured vertebra. LS instrumentation was hook fixation (claw hooks attached to second upper vertebra and infralaminar hooks attached to first upper vertebra) above and pedicle fixation (pedicle screws attached to first and second lower vertebrae) below the fractured vertebra. RESULTS: As a result, measurements of local kyphosis, sagittal index, and anterior vertebral height compression showed that the LS group had a better outcome at final follow-up (P < 0.05). Also, the SS group had a 55% failure rate, whereas the LS group had prolonged operative time and increased blood loss. However, there was no difference between the two groups according to Low Back Outcome Score. CONCLUSIONS: In conclusion, radiographic parameters demonstrated that LS instrumentation is a more effective management of thoracolumbar burst fractures. Nevertheless, clinical outcome was the same between the two groups. However, our conclusions were based on posterior-only surgery. Anterior column support would negate the need for LS fixation. Also, SS would have been more successful if two above and two below pedicle screws were used. PMID- 16306835 TI - Clinical results after cervical laminoplasty: differences due to the duration of wearing a cervical collar. AB - OBJECTIVE: This investigation assessed the relationship between the duration of wearing a cervical collar and clinical results in patients undergoing laminoplasty. METHODS: Twenty-five patients with cervical myelopathy who underwent laminoplasty and wore a cervical collar for 8 weeks postoperatively were followed for an average of 27.3 months. A second group of 26 patients who underwent laminoplasty and wore a cervical collar for 4 weeks was followed for an average of 32.8 months. RESULTS: In both groups, neurologic recovery rate and range of motion on lateral cervical radiographs were longitudinally compared with those obtained preoperatively. There was no significant difference in neurologic recovery between the two groups. In patients who wore the cervical collar for 4 weeks, the total range of the cervical spine was better maintained, especially the extension range of the cervical spine, compared with that in the patients who wore the cervical collar for 8 weeks. CONCLUSIONS: It is speculated that early removal of the cervical collar prevents contracture of the facet joint and postoperative atrophy and dysfunction of the extensor musculature of the cervical spine. PMID- 16306836 TI - Surgical outcome of ossification of the posterior longitudinal ligament (OPLL) of the thoracic spine: implication of the type of ossification and surgical options. AB - OBJECTIVE: Ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine produces myelopathy through anterior spinal cord compression that is usually progressive and unaffected by conservative treatment. Therefore, early decompressive surgery is imperative. However, decompression surgery of thoracic myelopathy is difficult, and the outcome is often poor. A retrospective study was conducted to investigate the surgical outcome of 21 patients with thoracic OPLL to evaluate which type of surgical approach is better and which type of thoracic OPLL results in a better surgical outcome. METHODS: A total of 21 patients with thoracic OPLL (10 men and 11 women; mean age 54 years), who underwent surgical treatment at our department from March 1985 to October 2000, were included in the study. Seven patients exhibited the flat-type OPLL and underwent either anterior decompression and fusion (one patient), anterior decompression via a posterior approach (three patients), or expansive laminoplasty (three patients). Fourteen patients exhibited the beak-type OPLL and also underwent either anterior decompression and fusion (two patients), anterior decompression via a posterior approach (six patients), or expansive laminoplasty (six patients). RESULTS: Regarding of operative time and blood loss, there were no marked differences between the two types of OPLL, regardless of the type of surgical procedure; anterior decompression and fusion and anterior decompression via a posterior approach yielded longer operative times and larger blood loss volumes than expansive laminoplasty. Concerning clinical outcome, there were five cases of neurologic deterioration. All of the five deteriorated cases were of the beak type OPLL treated by a posterior approach. Two of these patients were treated with expansive laminoplasty. CONCLUSIONS: There were five instances of neurologic deterioration in our thoracic OPLL series, and all of them exhibited beak-type OPLL. In the beak-type OPLL, a subtle alteration in the spinal alignment during posterior decompression procedures may increase spinal cord compression, leading to the deterioration of symptoms. A potential increase in kyphosis following laminectomy should be avoided by fixation with a temporary rod. If intraoperative monitoring suggests spinal cord dysfunction, an anterior decompression procedure should be attempted as soon as possible. PMID- 16306838 TI - Fractures and dislocations of the cervicothoracic junction. AB - Presented is a retrospective review of case notes and all available imaging studies in seven patients with acute fractures-dislocations of the cervicothoracic junction. Imaging studies included radiographs (five cases), computed tomography (six cases), and magnetic resonance imaging (seven cases). The study group consisted of five men and two women with mean age at presentation of 43.6 years (range 25-69 years). Four patients had been in road traffic accidents, whereas three patients had had falls. Three patients sustained complete neurologic deficits with no recovery, whereas the remaining four had no abnormal neurology or mild deficit at presentation and were normal at final follow-up. The injury was missed initially in three cases. The commonest injury pattern was traumatic spondylolisthesis of C7 on T1 with multilevel neural arch fractures, resulting in increased anteroposterior canal dimensions (four cases). Bilateral pars fractures of C7 and pure facet dislocation were seen in one case each. Neurologic deficit was related to the degree of anterior displacement of C7 on T1. Fracture-dislocation at the cervicothoracic junction is a rare injury with a variation of injury patterns and neurologic outcome. PMID- 16306839 TI - Biomechanical evaluation of the pullout strength of cervical screws. AB - OBJECTIVE: In the process of anterior cervical fusion, little is known about the biomechanics of anterior cervical screw pullout. In this study, three different aspects of cervical screw fixation were evaluated: self-tapping (ST) versus self drilling (SD) screws, the effect of screw geometry (length, diameter, thread pitch), and the use of rescue screws. METHODS: Nine screws consisting of different diameters, lengths, and thread pitch (cancellous and cortical) were tested in peak pullout force in an artificial bone model using an MTS 858 Mini Bionix test system. Rescue screws (4.5 mm) were then inserted in the failed holes of 4.0-mm screws and extracted to determine their holding strength. RESULTS: Length of screws and thread pitch both had a significant effect on the pullout force. Each 1 mm of increased screw length translates to 16 N of increased force to pullout in the foam bone model. Pullout strength did not vary significantly according to screw diameter or between SD and ST screws. However, the SD screw has an advantage because it can decrease the length of surgery. A decrease in pullout force of between 43% and 70% was found when using rescue screws. CONCLUSIONS: In situations in which the use of rescue/salvage screws is required, the surgeon should anticipate a significant decrease in the holding force compared with the original screw. Future directions for research include an evaluation of pullout force for screw and plate constructs. PMID- 16306840 TI - Evaluation of cyanoacrylate augmentation of transpedicular screw pullout strength. AB - PURPOSE: Pedicle screw fixation of osteoporotic bone in the elderly is a challenge. Various augmentation methods have been studied by many authors. Although polymethylmethacrylate (PMMA) augmentation is believed to be a standard method, its usage is fraught with complications. Butyl-2-cyanoacrylate is an alternative to PMMA as it is bioresorbable, biocompatible, inexpensive, and noninfective. The objective of the current study was to determine the pullout strength of the pedicle screws when butyl-2-cyanoacrylate is used for augmentation. METHODS: Fresh calf lumbar vertebrae were obtained from male calves weighing 100-120 kg and implanted with pedicle screws. The screws were placed in native, unaugmented bone (group 1), butyl-2-cyanoacrylate-augmented bone (group 2), and PMMA-augmented bone (group 3). Axial pullout tests were done by an Instron 4411 universal testing machine. Statistical analysis was performed using the SPSS 9.0 for Windows program. Paired samples t test was used, and P < 0.05 was considered significant. RESULTS: The mean bone mineral density of the vertebrae was 1.6 +/- 0.1 g/cm2. The mean pullout strengths were 1.55 +/- 0.23 kN for group 1, 1.62 +/- 0.42 kN for group 2, and 2.55 +/- 0.22 kN for group 3. There was no statistically significant difference between groups 1 and 2. PMMA augmentation increased the pullout strength significantly when compared with butyl-2-cyanoacrylate augmentation and native bone (P = 0.002 and P = 0.001, respectively). CONCLUSIONS: The results of this study show that butyl-2 cyanoacrylate has no contribution to the augmentation of pedicle screw fixation in a calf model when compared with native bone or PMMA augmentation. Further studies are required to evaluate the effectiveness of butyl-2-cyanoacrylate in osteoporotic specimens and under cyclic loading in calf vertebra and animal and cadaver models before dispensing with its utility as an augmentation method in the clinical setting. PMID- 16306841 TI - A prospective analysis of intraoperative electromyographic monitoring of posterior cervical screw fixation. AB - OBJECTIVE: This is a prospective study of 26 patients undergoing posterior cervical spine instrumentation with lateral mass or pedicle screws to determine the correlation between intraoperative screw stimulation thresholds and the position of posterior cervical lateral mass and pedicle screws. METHODS: One hundred forty-seven posterior cervical screws (122 lateral mass screws and 25 C7 pedicle screws) in 26 patients were electrically stimulated intraoperatively and stimulation thresholds recorded. Computed tomography (CT) scans were taken postoperatively and were evaluated independently to assess screw position. Electromyographic (EMG) thresholds and CT data were compared to assess the accuracy of the EMG screw stimulation technique in detecting screw malposition. RESULTS: Intraoperative electrical stimulation was accurate in verifying screw position. A stimulation threshold of 15 mA provided a 99% positive predictive value (89% sensitivity, 87% specificity) that the screw was within the lateral mass or pedicle. Stimulation values of 10-15 mA provided a 13% predictive value (66% sensitivity, 90% specificity) that the screw was within the lateral mass or pedicle. A stimulation value of <10 mA provided a 100% predictive value that the screw was malpositioned (70% sensitivity, 100% specificity). CONCLUSIONS: Intraoperative evoked EMG monitoring is a valuable tool in posterior cervical instrumentation using lateral mass and pedicle screws. Stimulation thresholds in this study correlated with screw position. Stimulation values of >15 mA reliably predict acceptable screw position. Values between 10 and 15 mA are generally associated with acceptable screw position, although exploration is recommended. Values below 10 mA are associated with screw malposition and warrant exploration, repositioning, and possible removal. PMID- 16306842 TI - Lateral lumbar disc herniations treated with a paraspinal approach: an independent assessment of longer-term outcomes. AB - In this independent review, at an average of 5.5 years after surgical excision of a lateral zone disc herniation using a paraspinal approach, 51 of 60 patients (85%) were satisfied with the results. Pain was eliminated in 36 (60%) and reduced in all but 3 (5%). Two-thirds of patients had no residual weakness or numbness. Thirty patients (50%) developed some new low-grade back pain, 17 (28%) developed some radicular pain, 9 (19%) developed some degree of radiographic instability, and 9 (15%) required further operative procedures. Based on this information, surgeons should be able to provide realistic expectations for their patients regarding outcomes. Based on our review of the literature, results using the paraspinal window appear similar, but not superior, to other available techniques, and the choice of approach should be based on the comfort of the surgeon. PMID- 16306843 TI - Accuracy and safety of thoracic pedicle screw placement in spinal deformities. AB - OBJECTIVES: To determine the safety of pedicle screw fixation in thoracic spine deformity correction. METHODS: One hundred twelve pedicle screws were surgically placed in 25 patients with degenerative, posttraumatic, and Scheuermann kyphosis and idiopathic and neuromuscular scoliosis. Screw position was evaluated using intraoperative and postoperative radiographs and thin-slice computed tomography. RESULTS: Of the total 112 thoracic pedicle screws that were inserted, 98 screws (87.5%) were fully contained within the cortical boundaries of the pedicle. When comparing proximal screws (T1-T8) with distal screws (T9-T12) and convex placed screws with concave ones, a statistically significant difference in screw placement was evident (P < 0.05). More misplaced screws were seen proximally and on the concave side. Of the 14 malpositioned screws, 2 (1.8%) demonstrated aortic abutment. There were no neurologic deficits, vascular injuries, or mechanical failures recorded. CONCLUSIONS: Placement of thoracic pedicle screws is both feasible and safe. PMID- 16306844 TI - High pulmonary risk scoliosis surgery: role of noninvasive ventilation and related techniques. AB - OBJECTIVE: There is little awareness among surgeons of the potential for noninvasive mechanical ventilation as an alternative to prolonged endotracheal intubation or tracheostomy for patients with neuromuscular scoliosis and ventilatory failure. These methods have not been reported for the perioperative management of scoliosis correction in patients with an inability to sustain their alveolar ventilation. METHODS: Five children with flaccid scoliosis secondary to muscular dystrophy or spinal muscular atrophy who had very high pulmonary risk were preoperatively trained in the use of noninvasive intermittent positive pressure ventilation (IPPV) and mechanically assisted coughing prior to spinal fusion. RESULTS: All patients were extubated by the third postoperative day to noninvasive IPPV despite continuous ventilator dependence. No patient developed any postoperative pulmonary complications or required a tracheotomy. CONCLUSIONS: It is critical for the orthopedic surgeon to be aware of these noninvasive options to tracheotomy to decrease the tendency to avoid surgery for these otherwise high-risk surgical patients. PMID- 16306845 TI - Late paraplegia after scoliosis treatment: an uncommon diagnosis. AB - We report the case of a young girl treated at age 16 for a progressive scoliosis by posterior instrumented arthrodesis. Ten years later, she suddenly developed lumbar pain and paraplegia. The surgical procedure showed a mass infiltrating the vertebral canal and the dural sheath following a supralaminar hook. Histology revealed a diagnosis of leiomyosarcoma. The outcome was poor with a rapid and fatal extension of the tumor. There is nothing to justify a causal link between the scoliosis and the late onset of a malignant tumor. Nevertheless, we discuss the potential role of diagnostic irradiation consecutive to scoliosis monitoring during growth and the potential role of environmental carcinogens like metallic biomaterials. Finally, rapid intrusion of this extraspinal tumor into the dural sheath resulted in a confusing clinical picture and delayed the diagnosis and treatment of the tumor. PMID- 16306846 TI - Monostotic fibrous dysplasia of the lumbar spine: case report and review of the literature. AB - Monostotic fibrous dysplasia of the spine is extremely rare. We present a 57-year old man who complained of persistent low back pain with monostotic fibrous dysplasia of the lumbar spine. Computed tomography revealed a lytic expansile lesion and marginal sclerosis in the L2 posterior element, although a bone scan did not reveal increased uptake in the lesion. The patient underwent total excision of the tumor via a posterior approach. Two years later, he is asymptomatic with no recurrence of the lesion, as confirmed by imaging. PMID- 16306847 TI - Clear cell meningioma of the cauda equina in an adult: case report and literature review. AB - In the pediatric population, clear cell meningiomas are more frequently intracranial than intraspinal in location. Tumors recur in up to 40% of cases within 15 postoperative months and are often managed with repeated resection with or without radiation therapy. The management strategy for adults with clear cell meningiomas involving the lumbar spinal canal (cauda equina) is less clearly defined. A 41-year-old woman presented with mild, right greater than left, lower extremity paresis. An enhanced magnetic resonance (MR) scan revealed a homogeneously enhancing intradural lesion filling the spinal canal at the L3-L4 level. Preoperative noncontrast MR studies of the brain and cervical and thoracic spine were negative. An L2-L5 laminectomy was performed for gross total excision of the intradural lesion, which was adherent to one nerve root of the cauda equina. Frozen-section diagnosis confirmed clear cell tumor. Differential diagnoses included meningioma versus renal cell carcinoma. Negative postoperative chest, abdominal, and pelvic computed tomography studies ruled out tumor of renal cell origin. Enhanced MR studies of the neuraxis proved negative. Consultations with multiple oncologists and radiation therapists recommended neither radiation nor chemotherapy following this initial surgery. She remains disease-free 1 year postoperatively. The high recurrence rate for clear cell meningiomas in children requires repeated tumor resection with or without secondary radiation therapy. Following gross total resection of lumbar tumors in adults, reserving radiation therapy for secondary recurrences provides optimal management. PMID- 16306848 TI - Dorsal extradural thoracic disc fragment: a diagnostic challenge. AB - Dorsal epidural migration of an extruded disc fragment is an infrequent event, especially in the thoracic spine. An uncommon case involving a 55-year-old man is presented, with a 1-month history of paraparesis and thoracolumbar pain. Magnetic resonance imaging demonstrated a dorsally located, extramedullary mass at the T10 T11 intervertebral level. The lesion was suspected to be a tumor. The patient underwent a T10-T11 laminectomy. Intraoperatively, an encapsulated mass of soft tissue adherent to the dural sac was found. The pathologic diagnosis was inflammatory tissue and disc material. Six months after the operation, the patient remained asymptomatic, and radiologic control showed no residual mass. Although rare, a sequestered disc fragment should be included in the differential diagnosis of an enhancing posterior extramedullary thoracic mass. Preoperative diagnosis of such pathology is difficult because the clinical signs and radiologic images may not entirely exclude other more common thoracic spinal lesions, especially tumors. PMID- 16306849 TI - Spinal epidural hematoma after removal of an epidural catheter: case report and review of the literature. AB - We report a case of spinal epidural hematoma after removal of an epidural catheter. The patient had no background of anticoagulant therapy or coagulopathy; sudden severe back pain occurred immediately after removal of the catheter. The chance of this occurring is estimated to be between 1:150,000 and 1:190,000. We studied 40 previous reports from 1952 to 2000, and we also investigated anticoagulant therapy and pathologic states, puncture difficulties and bleeding at the point of insertion, and its onset. In 23 cases (57.5%), anticoagulant therapy had been performed, and in 5 cases (12.5%), coagulopathy or liver dysfunction had been recognized. In 20 cases (50%), the initial symptoms were recognized within 24 hours after removal of the epidural catheter. Although spinal epidural hematoma is a very rare condition, it is a serious complication of continuous epidural anesthesia. PMID- 16306851 TI - Population pharmacokinetics of cyclophosphamide and its metabolites 4 hydroxycyclophosphamide, 2-dechloroethylcyclophosphamide, and phosphoramide mustard in a high-dose combination with Thiotepa and Carboplatin. AB - The anticancer prodrug cyclophosphamide (CP) is activated by the formation of 4 hydroxycyclophosphamide (4OHCP), which decomposes into phosphoramide mustard (PM). This activation pathway is inhibited by thiotepa. CP is inactivated by formation of 2-dechloroethylcyclophosphamide (2DCECP). The aim of this study was to develop a population pharmacokinetic model describing the complex pharmacokinetics of CP, 4OHCP, 2DCECP, and PM when CP is administered in a high dose combination with thiotepa and carboplatin. Patients received a combination of CP (1000-1500 mg/m/d), carboplatin (265-400 mg/m/d), and thiotepa (80-120 mg/m/d) administered in short infusions over 4 days. Twenty blood samples were collected per patient per course. Concentrations of CP, 4OHCP, 2DCECP, PM, thiotepa, and tepa were determined in plasma. Using NONMEM, an integrated population pharmacokinetic model was used to describe the pharmacokinetics of CP, 4OHCP, 2DCECP, and PM, including the already described processes of autoinduction of CP and the interaction with thiotepa. Data were available on 35 patients (70 courses). The pharmacokinetics of CP were described with a 2-compartment model, and those of 4OHCP, 2DCECP, and PM with 1-compartment models. Before onset of autoinduction, it was assumed that CP is eliminated through a noninducible pathway accounting for 20% of total CP clearance, whereas 2 inducible pathways resulted in formation of 4OHCP (75%) and 2DCECP (5%). It was assumed that 4OHCP was fully converted to PM. Induction of CP metabolism was mediated by 2 hypothetical amounts of enzyme whose quantities increased in time in the presence of CP (kenz=0.0223 and 0.0198 hours). Induction resulted in an increased formation of 4OHCP (approximately 50%), PM (approximately 50%), and 2DCECP (approximately 35%) during the 4-day course, and concomitant decreased exposure to CP (approximately 50%). The formation of 2DCECP was not inhibited by thiotepa. Apparent volumes of distribution of CP, PM, and 2DCECP could be estimated being 43.7, 55.5, and 18.5 L, respectively. Exposure to metabolites varied up to 9 fold. The complex population pharmacokinetics of CP, 4OHCP, 2DCECP, and PM in combination with thiotepa and carboplatin has been established and may form the basis for further treatment optimization with this combination. PMID- 16306852 TI - Hematocrit influences immunoassay performance for the measurement of tacrolimus in whole blood. AB - The comparison between the MEIA II and the EMIT assays for tacrolimus measurement and the interference by the hematocrit were evaluated in 93 samples from routine therapeutic monitoring at tacrolimus concentrations less than 9 microg/L (group A). Additionally, the incidence of false-positive results were determined in samples (n=46) from patients who were not receiving the drug (group B). In group A, no statistical differences were observed between the mean+/-SD values obtained by MEIA II (5.14+/-2.28 microg/L) and EMIT (4.61+/-1.79 microg/L). The correlation coefficient and the regression equation (95% CI) between both assays, were 0.761 and EMIT=1.088 (0.90, 1.35) MEIA II -0.38 (-1.65, -0.46), respectively. When the samples were stratified according to the hematocrit, the median differences between the methods (MEIA II minus EMIT) were as follows: hematocrit35%, 0.25 microg/L (P=0.02). In group B, false-positive results (above the detection limit) were observed in 63.04% of samples analyzed by MEIA II and in 2.17% of samples analyzed by EMIT. The median differences in apparent tacrolimus results were significantly higher in the samples with the lowest hematocrit: 2.2 microg/L, 1.4 microg/L, and 0.0 microg/L in samples with hematocrit35%, respectively. In conclusion, the differences in the tacrolimus results obtained by MEIA and EMIT assays were higher in samples from patients with hematocrit less than 25%, and the MEIA assay demonstrated a high incidence of false-positive results. PMID- 16306853 TI - Development and validation of a high-throughput assay for quantification of the proliferation inhibitor ABT-578 using LC/LC-MS/MS in blood and tissue samples. AB - We report here a specific, automated LC/LC-MS/MS assay for the quantification of ABT-578 in human and rabbit blood and rabbit tissues for drug-eluting stent development. After protein precipitation, samples were injected into the HPLC system and extracted online using a high flow of 5 mL/min. The extracts were then backflushed onto the analytic column. The [M+Na] of ABT-578 (m/z 988.6-->369.4) and its internal standard sirolimus (m/z 936.5-->409.3) were monitored. Extraction and analysis took 4 minutes. The assay was validated following the US Food & Drug Administration guidelines. Linearity was 0.025-25 ng/mL for most matrices. In human blood, interday accuracies were 81.8% (at 0.025 ng/mL), 91.0% (1 ng/mL), and 99.5% (50 ng/mL), and interday precisions were 10.7% (0.025 ng/mL), 3.0% (1 ng/mL), and 1.8% (50 ng/mL). PMID- 16306854 TI - Steady-state pharmacokinetics and tolerability of indinavir-lopinavir/r combination therapy in antiretroviral-experienced patients. AB - Six HIV-positive antiretroviral experienced patients initiating therapy with a regimen including lopinavir/ritonavir (400/100 mg twice per day) and indinavir (800 mg twice per day) underwent steady-state pharmacokinetic analysis. The AUC0 12 h of indinavir when combined with lopinavir/ritonavir was comparable with previously published data on indinavir/ritonavir 800/100 mg twice per day in HIV infected individuals. However, lopinavir AUC0-12 h, Cmax, and C12 h were lower than previously reported in the absence of indinavir. The regimen was well tolerated, although 2 patients developed grade 3 hypertriglyceridemia. No patient discontinued the regimen because of indinavir-related urologic or retinoid-type adverse effects. Further study of the regimen with larger cohorts of patients is necessary. PMID- 16306855 TI - No significant influence of saquinavir hard-gel capsule administration on pharmacokinetics of lopinavir in combination with ritonavir: a population approach. AB - The influence of saquinavir hard-gel capsules on lopinavir pharmacokinetic parameters was investigated using a population approach. Forty-nine patients infected with human immunodeficiency virus type 1 and treated with lopinavir/ritonavir, nucleoside/nucleotide reverse transcriptase inhibitors plus saquinavir (group A), and 118 patients treated with lopinavir/ritonavir plus nucleoside/nucleotide reverse transcriptase inhibitors (group B) were included in the study. No significant relationship was established between the presence or the daily dosage of saquinavir in the treatment and lopinavir population pharmacokinetic parameters. The values (mean+/-standard deviation) of the individual apparent clearance (5.0+/-1.8 vs. 5.0+/-3.2 L/h), volume of distribution (66.6+/-1.6 vs. 66.8+/-1.9 L), absorption rate constant (0.37+/-0.01 vs. 0.37+/-0.03 hours), and trough plasma concentration (5.5+/-2.3 vs. 5.3+/-1.9 mg/L) of lopinavir are not significantly different between groups A and B. This lack of influence of saquinavir on lopinavir pharmacokinetics makes the use of this combination in salvage therapy easier. PMID- 16306856 TI - Pharmacokinetic behavior of rituximab: a study of different schedules of administration for heterogeneous clinical settings. AB - This study was designed to report the pharmacokinetic behavior of Rituximab in patients affected with different diseases and treated with different schedules of administration. A low tumor burden was a common feature of all patients (N=48) included in our study, whereas the timing of Rituximab administration varied from weekly (groups 1, 2, 3) to monthly (group 4). Group 1 included patients with follicular lymphoma treated with 4 weekly doses of Rituximab after first-line chemotherapy with CHOP. At the start of Rituximab, patients were in partial or complete clinical response but showed persistence of disease at molecular level (bcl-2-positive) in bone marrow and/or peripheral blood. Patients in group 2 had autoimmune disorders and Rituximab was given to act on B-cells, interfering with their production of autoantibodies. In patients with amyloidosis (group 3), Rituximab was given to kill progenitor B-cells of the small clone terminating in amyloid-producing plasma cells. In groups 2 and 3, the target of monoclonal antibody was a population of small B cells, which make an intrinsic feature of the diseases. Group 4 included patients with relapsed or refractory follicular and mantle cell lymphoma who underwent a salvage program of immunochemotherapy, purging in vivo and autotransplant: the first of the six planned doses of Rituximab was administered after a debulking phase with a third-generation regimen, such as VACOP-B. An enzyme-linked immunoassay (ELISA) developed and validated in our laboratory was used for the pharmacokinetic study. Rituximab disposition was characterized by a 2-exponential decay, with a long elimination half-life of approximately 3 weeks (range, 248-859 hours). The total systemic clearance ranged between 3.1 and 11.9 mL/hr/m. After 4 weekly infusions, Rituximab concentration was approximately 2.5 microg/mL, which is approximately 85% of the steady-state level. Steady-state plasma concentrations of Rituximab were reached after 6 to 8 weekly infusions. The adopted pharmacokinetic model (2 compartment open model) seems to provide the best fit of Rituximab disposition both during and after treatment, even when different schedules of drug administration are used. Because several studies reported an association between response and serum Rituximab concentrations, a treatment based on a pharmacokinetic model may be useful for predicting the desired drug concentration. PMID- 16306857 TI - Analysis for identification in amnesty bin samples from dance venues. AB - The analysis of unknown substances discarded in amnesty bins, first described by Ramsey et al, from a large central London club and 7 smaller clubs in Manchester, UK are described. The contents of the bins were collected between July 2003 and March 2004. Solid dosage formulations were identified using the TICTAC database, chemical tests, and GC-MS screening. Drugs that could not be readily identified were subjected to other analytical techniques. The goal was to document the current range of drugs available on the dance scene and compare the findings between the London club, which had been the subject of a previous survey, and Manchester clubs. More than 1000 tablets, capsules, and powder doses were discarded in the amnesty bins. Tablets containing only MDMA (ecstasy) were found to be >94% and >84% of the total in London and Manchester, respectively. Although the quantities of tablets and powders recovered were different between London and Manchester, the proportions of the drugs were remarkably similar. The most common drugs found in powders in London and Manchester respectively were cocaine (29%, 40%), amphetamine (25%, 26%), ketamine (19%, 20%), and MDMA (19%, 11%). PMID- 16306858 TI - Randomized, double-blind, placebo-controlled study about the effects of cannabidiol (CBD) on the pharmacokinetics of Delta9-tetrahydrocannabinol (THC) after oral application of THC verses standardized cannabis extract. AB - Cannabidiol (CBD) is known to modify the effects of Delta-tetrahydrocannabinol (THC) by decreasing anxiety and antagonizing other THC-effects. As a reason, pharmacodynamic as well as pharmacokinetic mechanisms were suggested. In context of the use of cannabis-based medicine extracts for therapeutic purposes, a study was performed in a double-blind and placebo-controlled cross-over design in which each of 24 volunteers (12 male and 12 female, age 18-45 years) obtained soft gelatin capsules with 10 mg THC (THC-set), cannabis extract containing 10 mg THC +5.4 mg CBD (CAN-set) or placebo in weekly intervals. Blood samples were taken 30 minutes before and 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 9 hours and 24 hours after the intake. The concentrations of THC, of its metabolites 11-OH-THC, THC-COOH and of CBD in the plasma samples were determined by automatic solid phase extraction, derivatization with N,O bis(trimethylsilyl)triflouroacetamide and gas chromatography-mass spectrometry. The concentration versus time curves (maximum concentrations Cmax, corresponding time tmax and areas under the curves AUC) were evaluated by statistical methods with respect to equivalence or differences between the CAN-set and the THC-set. Furthermore, the intra-individual ratios of Cmax and AUC for 11-OH-THC/THC, THC COOH/THC and THC-COOH/11-OH-THC were compared between the THC-set and the CAN set. Despite the large variation of the data, evidence emerged from the total of the results that CBD partially inhibits the CYP 2C catalyzed hydroxylation of THC to 11-OH-THC. The probability for this inhibition is particularly high for oral intake because THC and CBD attain relatively high concentrations in the liver and because of the high first-pass metabolism of THC. However, the effect of CBD is small in comparison to the variability caused by other factors. Therefore, a pharmacokinetic reason for the differences determined between pure THC and cannabis extract is improbable at the doses chosen in this study. Significantly higher AUC and Cmax and shorter tmax were found for females as compared with males. PMID- 16306859 TI - Baseline measures of fatty acid ethyl esters in hair of neonates born to abstaining or mild social drinking mothers. AB - Elevated levels of fatty acid ethyl esters have been documented in the meconium of neonates born to heavily drinking mothers. Recently, accumulation of FAEE has been documented in the hair of drinking adults. To be able to use this test in the diagnosis of Fetal Alcohol Spectrum Disorder, one needs to establish baseline FAEE levels in infants born to nonalcoholic women because ethanol occurs in the baby naturally even without drinking. Scalp hair of 56 infants born to nonalcoholic women attending a well-baby clinic was obtained. Levels of FAEE were measured by GC-MS. FAEE were detected in almost all hair samples. Offspring of women admitting to mild, infrequent drinking did not differ in FAEE hair levels from infants of total abstainers. Baseline levels of FAEE in neonatal hair of nonalcoholic mothers have been established. Mild infrequent maternal drinking does not elevate these baseline levels. This may help avoid false-positive determinations when assessing infants born to problem drinking mothers. PMID- 16306860 TI - Effect of hemodialysis on the metabolic clearance of 5-Fluorouracil in a patient with end-stage renal failure. AB - A 64-year-old man on chronic hemodialysis for end-stage renal disease developed peritoneal carcinomatosis, and palliative chemotherapy with fluorouracil was started. The drug was administered (325 mg/m as IV bolus, at 2 PM) on 2 separate occasions, ie, 1 hour after dialysis and 2 days later, 49 hours after dialysis. The time course of the fluorouracil plasma concentration was determined, and the main pharmacokinetic parameters were calculated. The slope of the monoexponential decay of plasma concentration was significantly greater 1 hour (0.161 minutes) than 49 hours after dialysis (0.127 minutes), and plasma clearance was correspondingly higher (1.78 L/min versus 1.46 L/min). The volume of distribution did not change (11.1 L versus 11.5 L). Because fluorouracil is minimally excreted by the renal route (about 10% of the dose) and is almost entirely metabolized by dihydropyrimidine dehydrogenase (DPD), it is suggested that plasma factors that accumulate during the interdialytic period and are removed by dialysis may inhibit DPD activity and, consequently, fluorouracil metabolic clearance. PMID- 16306861 TI - Differences in CYP3A5*3 genotype distribution and combinations with other polymorphisms between Spaniards and Other Caucasian populations. AB - The goal of this study was to detect genotypic differences between Spaniards and other related populations regarding CYP3A4*1B, CYP3A5*3, and ABCB1 (MDR1) C3435T polymorphisms. DNA from 177 Spanish patients were analyzed for the presence of these mutations using PCR-restriction fragment length polymorphism or direct sequencing. The observed frequencies for CYP3A4*1B, CYP3A5*3, and C3435T alleles were within normal values in Caucasians (0.04, 0.91, and 0.5, respectively). However, 2.8% of the patients were homozygous for the wild-type CYP3A5*1 allele, an extremely uncommon genotype in other Caucasians. In addition, analysis of CYP3A4-3A5 haplotypes revealed the existence of 2 unusual subgroups: patients who were homozygous wild-type for both polymorphisms, and patients showing a CYP3A4*1A/*1B-CYP3A5*3/*3 genotype combination. The incidence of CYP3A5*1/*1 carriers and the occurrence of subjects combining the 2 above-mentioned unusual genotype combinations were more frequent in Spanish-Caucasians compared with American- or European-Caucasians. ABCB1 C3435T genotype frequencies were equally distributed between both single and combined CYP3A4 and 3A5 genotypes. These findings suggest that dose requirements for drugs metabolized by CYP3A and certain allele-disease association studies in white populations could show discrepancies in Spaniards. PMID- 16306862 TI - Possible reduction in indinavir serum concentrations by bosentan. PMID- 16306863 TI - Menopause and disorders of the central nervous system. AB - The cessation of ovarian estrogen production occurring around the time of menopause has the potential to influence central nervous system function, as well as a number of neurological disorders that affect women during midlife and old age, including memory loss and mild cognitive impairment, ischemic stroke, Parkinson's disease, and Alzheimer's disease. During midlife, there is observational evidence that episodic memory is not substantially affected by natural menopause or by use of estrogen-containing hormone therapy, but short term clinical trial evidence suggests hormone therapy might benefit verbal memory after surgical menopause. Clinical trial data indicate that hormone therapy does not reduce, and may increase, stroke incidence. Parkinson's disease and Alzheimer's disease are the 2 most common neurodegenerative illnesses. Estrogen influences dopaminergic pathways within the central nervous system. However, available observational evidence is limited and inconclusive regarding any role of hormone therapy in influencing risk or symptoms of Parkinson's disease, a disorder of dopaminergic neurons. Finally, clinical trial data indicate that hormone therapy should not be initiated in the late postmenopause with the goal of improving memory, preventing cognitive decline, reducing dementia risk, or improving Alzheimer's disease symptoms. An important priority for clinical investigation is to determine whether hormone therapy used during the menopausal transition and early postmenopause has long-term effects on cognition or dementia risk. The critical window hypothesis as applied to Alzheimer's disease conjectures that effects of early hormone therapy might differ from those of hormone therapy initiated in the late postmenopause, but convincing evidence is yet to be obtained. PMID- 16306864 TI - [Urinary incontinence in postmenopausal period: clinical and pharmacological treatments]. AB - Urinary incontinence is a common clinical problem in female sex and occurs especially in postmenopausal women; this disease, that represents an economical problem for society, begins in young age, arises in middle age and increases in women more than 65 years old. Studies carried out on etiological factors involved in urinary incontinence show that estrogens enhance the trophism and vascularization of the muscular and fascial support of the pelvic floor, the growth of fibroblasts and the collagen metabolism in the superficial fascia in postmenopausal women. The postmenopausal estrogenic deficit could be related to many urogenital problems, but many researches performed on the effects of estrogens in urogenital postmenopausal homeostasis and of hormonal replacement therapy in postmenopausal incontinent women, did not show conclusive findings; for this reason, even if many authors attributed to menopause a role of major risk factor for incontinence, a direct correlation has never been confirmed. The treatment of postmenopausal female incontinence may be clinical and pharmacological, and includes a first step therapy (bladder training, biofeedback techniques, electrical pelvic floor stimulation) and a second step therapy (pharmacological therapy, bladder devices and surgical operations). In this review the clinical and pharmacological treatments, their efficacy and their application in incontinent postmenopausal women are described. PMID- 16306865 TI - Sex hormones in the regulation of bone and cartilage metabolism: an old paradigm and a new challenge. AB - The effects of estrogen on tissues such as bone, endometrium and breast have been extensively studied, and the pleitropic effects of the female sex hormone are well established. Cartilage is not generally viewed as an estrogen responsive tissue. However, several epidemiological studies, and a few recent intervention studies supports that estrogen may have a role in osteoarthritis (OA), and recent animal studies further suggests that estrogen may be involved in regulation of cartilage turnover. Accordingly the issue of chondroprotrective properties of estrogen has received increased attention in recent scientific publications. In this review, we summarize current studies indicating a role for estrogen in the regulation of cartilage turnover and development of joint diseases. We report results from in vitro and animal studies where the effects of ovariectomy and treatment with estrogen and selective estrogen receptor modulators (SERM) on cartilage erosion have been evaluated. Furthermore, we report results from assessment of the effects of estrogen and SERM in postmenopausal women which shed new light on the interactions between estrogen and joint tissues. It still remains to be established whether estrogen or SERM could find a role in prophylaxis and/or treatment of OA, and much work lies ahead. Current data reviewed in this manuscript can be considered encouraging and they raise the hope that new treatment options for OA may become available based on estrogen and, in particular, compounds acting through the estrogen receptor. However, at present hormone replacement therapy and SERMs available in clinical practice, cannot be recommended as a therapy for arthritic disease. PMID- 16306866 TI - [Hyperhomocysteinemia: associated obstetric diabetes and fetal malformations]. AB - In its biological complexity, pregnancy represents a challenge both for the maternal organism and the fetal development and growth. During this period, some peculiar pathologies of pregnancy can occur which can involve or the fetus only i.e.: spontaneous pregnancy loss, intrauterine growth retardation, defects of neural tube, until the intrauterine fetal death; or pathologies occurring in the placenta and thus involving maternal organism and fetus too, such as pre eclampsia. All these pathologies recognize many risk factors, among them the hyperhomocysteinemia. Hyperhomocysteinoemia can be caused by enzymatic defects or lack of some vitamins cofactors (vitamin B6, vitamin B12 and folic acid). The genetic defects which, as homozygous genotype, cause high plasma levels of homocysteine are already well known; they lead to an activity reduction of the enzymes responsible for their metabolism, for example: the deficiency of cystathionine beta-synthase; the deficiency of the methylcobalamine production; the deficit of the 5-10 methylenetethrahydrofolate reductase (MTHFR). However, even the heterozygous genotypes, which have a variable incidence from 1/70 to 1/200 and directly of 5-15% for the C677T mutation of the 5-10 MTHFR, can determine a mild hyperhomocysteinemia with a consequent cardiovascular risk. The close implications, widely demonstrated in the international literature, between hyperhomocysteinemia and the maternal-fetal diseases are described. PMID- 16306867 TI - HIV-HCV co-infection during pregnancy. AB - AIM: Hepatitis C virus (HCV) infection is common in human immunodeficiency virus (HIV) infected individuals in the United States and in Western Europe. The aim of this study is to describe our experience in the management of HIV-HCV co-infected women during pregnancy and to report the rates of HIV and HCV vertical transmission in this particular population. METHODS: This is a prospective study including all HIV-HCV coinfected women who delivered in the major public hospital of Genoa from April 1990 to December 2002. The method of delivery and breastfeeding habits were recorded. The babies were monitored at the paediatric referral centre. RESULTS: Thirty-nine HIV-HCV co-infected pregnant women were included in the analysis, 2 of these women delivered twice during the study period. The mean age (+/-SD) of this population at delivery was 30.4+/-4.9 years. Thirty-four (87.2%) HIV-HCV co-infected women had a history of intravenous drug use. Seventeen (43.5%) women reported regular alcohol intake during pregnancy. Forty-one infants were born during the study period. There was no case of HIV vertical transmission in our population. Thirty-six (87.8%) of 41 babies were born to women with positive HCV-RNA. Two babies (4.9%) were HCV-infected. CONCLUSIONS: The improved prognosis for HIV-infected people has increased the desire of parenthood. There is no evidence to support advising against pregnancy in HIV-HCV co-infected women. PMID- 16306868 TI - [Preliminary experience with transobturator tape for treatment of urinary incontinence in women]. AB - AIM: Aim of this study is to report our preliminary experience with a minimally invasive surgical procedure using transobturator approach. METHODS: We enrolled in our study 19 patients with stress urinary incontinence and urethral hypermobility without genital prolapse. All patients underwent suburethral transobturator tape (TOT) procedure. Four patients underwent previous surgical procedure for incontinence (3 Kelly and 1 Burch). No patients reported previous major gynaecological surgery. Mean age was 58.68 years (range 36-75). The sling was placed according to the technique described by Delorme. RESULTS: Mean operating time was 21 min (range 14-48). No bladder lesions or intraoperative complications occurred. Fifteen patients were dismissed the same day of surgery. Mean hospital staying was 1.4 days (range 1-5). No infections, erosions or sieromas have been observed. CONCLUSIONS: Our experience shows as TOT technique is extremely rapid, reproducible and with a very short learning curve. The transobturator approach avoids the risk of bladder, bowel or vascular injuries. Compliance and patient satisfaction have been very encouraging, restoring a real functional well-being. However, the small number of patients in our study and the short follow up do not actually allow us to draw definitive results and further studies are needed to confirm the technique success. PMID- 16306869 TI - [Sonoembryological criteria in the evaluation of gestational age]. AB - AIM: The progressive appearance of specific embryological structures has been successfully used to define the ultrasonographic gestational age (GA). In our study we have revised this technique by comparing our results with the ones from Warren et al. obtained in 1989. Aim of the study is to evaluate the accuracy of this technique when applied working with new instrumentations and average skilled sonographers. METHODS: Sixty-five pregnant women with correct GA ranging between 4 and 12 weeks have been observed between April 2003 and March 2004. We exposed them to an ultrasound examination looking for these structures in terms of presence/absence: gestational sac, yolk sac, embryo pole with cardiac activity, thromboencephalic cavity, falx cerebri and physiologic midgut herniation. We used transvaginal sonography (TVS) until 11 weeks of GA, from 11 weeks + 1 day we switched to transabdominal sonography (TAS). We have compared our results with the ones published by Warren et al. RESULTS: Gestational sac's visualization has been possible between 4 weeks+3 days/5 weeks of GA. Yolk sac has been visualized between 5 weeks+4 days/6 weeks of GA, embryo with cardiac activity has been observed between 5 weeks+6 days/6 weeks+2 days of GA, romboencephalic cavity has been visualized between 7 weeks and 7 weeks+5 days, falx cerebri has been detected between 9 weeks+1 day/10 weeks+3 days. Visualization of physiologic midgut herniation has been possible between 8 weeks+3 days/8 weeks+6 days; it disappeared between 10 weeks+3 days/11 weeks +1 day. CONCLUSIONS: Although the technological improvement in the US equipment in the last 15 years and the combination of TVS with TAS, there has been no advanced visualization of those single embryological structures. Indeed there has been a reduction of the time range during which those structures have been visualized. This improvement might have important practical implications in the ultrasonographic assessment of GA. PMID- 16306870 TI - Prenatal diagnosis of placental chorioangioma: our experience. AB - Placental chorioangioma is the most common benign tumor of placenta. The relationship of vascularized chorioangiomas to adverse pregnancy outcome is well recognized. We report 3 cases of placental chorioangioma. Hypervascularization of the lesions in all patients and the immune hydrops with adverse fetal outcome in 2 cases are the complications of our mini-series. Ultrasonography and Doppler ultrasonography findings were useful in establishing the prenatal diagnosis and the prognosis. PMID- 16306871 TI - [Voluminous ectopic polycystic ovaries in Mayer-Rokitansky-Kuster-Hauser syndrome]. AB - We report the case of a 20-year-old woman with primary amenorrhea, normally developed sexual characteristics, infantile external genitals and absence of vaginal orifice. Blood chemistry studies showed elevated serum levels of gonadotropin, estrogens, testosterone, 17OH-progesterone, prolactin and TSH and low concentrations of FT(3) and FT(4). Cytogenetic karyotyping and in situ fluorescence hybridization revealed a 46,XX karyotype and the presence of Barr chromatin body. Laparoscopy disclosed absence of a vagina, apparently normal, enlarged tubes, multifolicular ectopic gonads and symmetric bilateral uterine buds. Diagnostic work-up included skeletal radiography, renal ultrasonography, intravenous pyelography, pelvic echography, pelvic phlebography and magnetic resonance imaging studies to demonstrate possible associations between the genitourinary and skeletal anomalies. Based on the clinical and laboratory findings, a diagnosis of Mayer-Rokitansky-Kuster-Hauser syndrome was established, associated with the presence of enlarged ectopic polycystic ovaries in the abdominal cavity. PMID- 16306872 TI - Dairy consumption and ovarian cancer risk in the Netherlands Cohort Study on Diet and Cancer. AB - Ovary cancer risk in relation to consumption of dairy products was investigated using a self-administered questionnaire on dietary habits and other risk factors for cancer, which was completed in 1986 by 62 573 postmenopausal women participating in the Netherlands Cohort Study. Follow-up for cancer was implemented by annual record linkage with the Netherlands Cancer Registry and a nationwide pathology registry. After 11.3 years of follow-up, data of 252 incident epithelial ovarian cancer cases and 2216 subcohort members were available for analysis. No association was seen between consumption of milk, yoghurt, cheese or fermented dairy products and ovarian cancer risk. The multivariable adjusted relative risk of epithelial ovarian cancer for women in the highest compared to the lowest quintile of intake of lactose or dairy fat was 0.93 (95% confidence interval (CI)=0.60-1.45; P(trend)=0.32) and 1.53 (95% CI=1.00-2.36; P(trend)=0.11), respectively. Lactose or dairy fat intakes were not associated with serous ovarian cancer risk. Our results do not support an association between consumption of dairy products or lactose intake and ovarian cancer. PMID- 16306873 TI - Combination chemotherapy with carboplatin, capecitabine and epirubicin (ECarboX) as second- or third-line treatment in patients with relapsed ovarian cancer: a phase I/II trial. AB - Platinum-based combination chemotherapy has been proven to be superior to single agent platinum in the treatment of relapsed ovarian cancer after a treatment-free interval of more than 6 months. A response rate of 41% was previously reported by our group using a combination of epirubicin, cisplatin and 5-FU in patients who relapsed within 12 months, we therefore assessed a similar, but more convenient combination of epirubicin, carboplatin and capecitabine in this phase-I/II trial. In total, 18 patients with recurrent epithelial ovarian carcinoma, who had not received more than two lines of chemotherapy and the treatment-free interval exceeded 6 months were treated with carboplatin AUC5, epirubicin 50 mg m(-2) and capecitabine at several dose levels on continuous 21 day cycles and 14 of 21 day cycles. Patients were assessed for toxicity and by CT and CA-125 for response. The overall response rate was 61.1%, with three complete and eight partial responses. Grade 3/4 haematological toxicity was seen in 10 out of 18 patients and caused dose reductions and treatment delays. The combination of epirubicin, carboplatin and capecitabine showed good activity but caused excessive toxicity. A phase-II trial using carboplatin and capecitabine is underway. PMID- 16306874 TI - Birth outcome in women with breast cancer. AB - We investigated whether maternal breast cancer affects birth outcome in a nationwide cohort study of 695 births from 1973 to 2002 of women with breast cancer with respect to preterm birth, low birth weight at term, stillbirth and congenital abnormalities as well as mean birth weight, compared with the outcomes of 33 443 births from unaffected mothers. There was no excess risk of adverse birth outcome for the 216 newborns of women with breast cancer before pregnancy. Stratification by mother's treatment did not change the results. For 37 newborns of women diagnosed during pregnancy, the prevalence ratio (PR) of preterm birth was 8.1 (95% confidence interval (CI): 3.8-17). However, 10 of the 12 preterm deliveries among these women were elective early deliveries. Among 442 births of women diagnosed in the 2 years from time of delivery, the PR of preterm birth was 1.4 (95% CI: 1.0-2.0), and the PR of low birth weight at term for boys was 2.9 (95% CI: 1.3-6.3). Overall, our results are reassuring regarding the risks of adverse birth outcome for breast cancer patients. PMID- 16306875 TI - Levels of omega-3 fatty acid in serum phospholipids and depression in patients with lung cancer. AB - Previous studies suggested that omega-3 fatty acids (FAs) have therapeutic effects against depression, but there is no evidence in the oncological setting. Our preliminary study reported the association between lower omega-3 FA intake and occurrence of depression in lung cancer patients. To explore the association further, the present study examined whether depression was associated with lower levels of omega-3 FAs in serum phospholipids. A total of 717 subjects in the Lung Cancer Database Project were divided into three groups by two cutoff points of the Hospital Anxiety and Depression Scale depression subscale (HADS-D). In all, 81 subjects of the nondepression and minor depression groups (HADS-D<5 and 510) for age, gender, clinical stage, and performance status. Fatty acids were assayed by gas chromatography and compared among the three matched groups. There were no differences between the major depression group and nondepression group in any FAs. The minor depression group had higher mean levels of docosahexaenoic acid (mean+/-s.d. (%), nondepression: 7.40+/-1.54; minor depression: 7.90+/-1.40; major depression: 7.25+/-1.52, P=0.017). These results suggested that serum FAs are associated with minor, but not major, depression in lung cancer patients. PMID- 16306876 TI - A twist for survival and cancer progression. AB - A major obstacle to the expansion of abnormal cells with significant proliferative potential is the induction of programmed cell death. Consequently, oncogene-driven hyperproliferation must be associated with apoptosis inhibition to allow malignant outgrowth. The oncogenic cooperation of N-Myc and Twist-1 in the development of neuroblastoma, the most common and deadly solid tumour of childhood, perfectly illustrates such a process. N-Myc promotes cell proliferation, whereas Twist-1 counteracts its pro-apoptotic properties by knocking-down the ARF/p53 pathway. On the basis of numerous recent studies reporting its overexpression in a variety of human cancers, we discuss in this review the role of Twist-1 as a potent inhibitor of the cell safety programs engaged in response to an abnormal mitogenic activity. PMID- 16306877 TI - Epidermal growth factor receptor dimerization status determines skin toxicity to HER-kinase targeted therapies. AB - Skin toxicity, a common drug-related adverse event observed in cancer patients treated with epidermal growth factor receptor (EGFR)-directed therapies is rarely seen with therapies targeting HER2. This study reports the significance of the EGFR and HER2 dimerization status in skin with regard to these dermatologic side effects. We demonstrate the differential effect of HER-directed therapies on the ligand driven activation status of EGFR, HER2 and MAPK in normal human epidermal keratinocytes. EGFR-directed therapies, such as gefitinib and cetuximab, inhibited ligand-induced activation of EGFR and MAPK in human keratinocytes. Pertuzumab, an antibody interfering with functional HER2 heterodimerization, failed to block ligand-induced HER signaling in primary keratinocytes. Using a novel proximity-based dimerization assay (eTagtrade mark) we show that EGFR homodimers are the predominant HER dimer pair in normal primary kertinocytes and in normal skin tissue from 16 patients with solid malignancies. The presence of [p]EGFR and [p]MAPK, but the absence of [p]HER2, demonstrates productive signaling via EGFR but not HER2 in human skin. These data illustrate the importance of the EGFR dimerization partner in human skin and suggests that inhibition of EGFR homodimer signaling rather than EGFR/HER2 heterodimer signaling maybe the key molecular event determining dermatologic toxicity discrepancies observed between EGFR-targeted versus HER2-targeted therapies. PMID- 16306878 TI - Segmental duplication density decrease with distance to human-mouse breaks of synteny. AB - Segmental duplications are large genomic segments of recent origin and nearly identical sequence. Segmental duplications account for up to 5% of the human genome and they are often involved in genomic rearrangements and human disease. We developed a rapid computational method to characterize segmental duplications in the mouse and the human genomes according to four sequence assemblies for each species. Segmental duplication content in the mouse genome assemblies has largely changed over the four releases (from 0.2 to 1.2%, 4.5 and 3.0%), while in the four human assemblies duplication content was 4.8, 3.5, 3.7 and 3.7%, respectively. This suggests that cataloguing and assembling duplications has been challenging in both genomes and any interpretation of comparative analyses of duplication content must keep this in perspective to avoid artifacts. Human and mouse segmental duplications are more frequent than expected in regions where there is a syntenic discontinuity and the duplication content in syntenic regions decreases significantly with distance from breakpoints of synteny. These observations indicate that in mouse and human the frequency of segmental duplications is strongly correlated with distance to human and mouse syntenic breaks or the most dynamic regions in evolution.. PMID- 16306879 TI - Do we need a uniform regulatory system for biobanks across Europe? AB - Within Europe, there is currently no uniform regulatory system that applies to human biobanks used for genetic research purposes. This has resulted in considerable variation in the national law that applies to the use of DNA samples, personal information and medical records in the countries across Europe. This could result in a situation where researchers collaborating across Europe may be operating unlawfully if they share research data and samples across borders where different laws are in operation. There are also concerns that the lack of standardised guidelines inhibits cooperation among researchers across Europe but also restricts the sharing of DNA samples and information across national borders, which is problematic for multinational companies and institutions carrying out collaborative research. Ultimately, the lack of a uniform regulatory system may have implications for the viability and long-term competitiveness of collaborative European research. The purpose of this paper is to discuss some of the preliminary issues that would need to be considered before such a regulatory system for biobanks could be developed within Europe. PMID- 16306880 TI - Optimal genotype determination in highly multiplexed SNP data. AB - High-throughput genotyping technologies that enable large association studies are already available. Tools for genotype determination starting from raw signal intensities need to be automated, robust, and flexible to provide optimal genotype determination given the specific requirements of a study. The key metrics describing the performance of a custom genotyping study are assay conversion, call rate, and genotype accuracy. These three metrics can be traded off against each other. Using the highly multiplexed Molecular Inversion Probe technology as an example, we describe a methodology for identifying the optimal trade-off. The methodology comprises: a robust clustering algorithm and assessment of a large number of data filter sets. The clustering algorithm allows for automatic genotype determination. Many different sets of filters are then applied to the clustered data, and performance metrics resulting from each filter set are calculated. These performance metrics relate to the power of a study and provide a framework to choose the most suitable filter set to the particular study. PMID- 16306881 TI - Haplotype analysis of tumour necrosis factor receptor genes in 1p36: no evidence for association with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with partially understood aetiology. The 1p36 region has been previously linked with SLE and harbours tumour necrosis factor receptor (TNFR) genes. Functional and genetic data implicate their gene products in SLE and other autoimmune diseases. In all, single-nucleotide polymorphisms (SNPs) across TNFRSF14 (HVEM), and 43 SNPs across the TNFRSF8 (CD30) and TNFRSF1B (CD120B) locus were investigated for linkage disequilibrium (LD) and haplotype analysis in European-Caucasians. Strong LD was observed across HVEM and CD120B, and little LD and recombination across CD30. We also examined the association of SNPs and haplotypes in HVEM, CD30 and CD120B with SLE in European-Caucasians. There was no evidence of association for these genes in 456 European-Caucasian families with SLE from UK. Haplotype tagging SNPs are made known across areas of strong LD, which will facilitate analysis for susceptibility in other diseases. PMID- 16306882 TI - Clustering of haplotypes based on phylogeny: how good a strategy for association testing? AB - Haplotypes are now widely used in association studies between markers and disease susceptibility locus. However, when a large number of markers are considered, the number of possible haplotypes increases leading to two problems: an increased number of degrees of freedom that may result in a lack of power and the existence of rare haplotypes that may be difficult to take into account in the statistical analysis. In a recent paper, Durrant et al proposed a method, CLADHC, to group haplotypes based on distance matrices and showed that this could considerably increase the power of the association test as compared to either single-locus analysis or haplotype analysis without prior grouping. Although the authors considered different one-disease-locus susceptibility models in their simulations, they did not study the impact of the linkage disequilibrium (LD) pattern and of the susceptibility allele frequency on their conclusions. Here, we show, using haplotype data from five regions of the genome of different lengths and with different LD patterns, that, when a single disease susceptibility locus is simulated, the prior grouping of haplotypes based on the algorithm of Durrant et al does not increase the power of association testing except in very particular situations of LD patterns and allele frequencies. PMID- 16306883 TI - Nematodes learn: now what? PMID- 16306884 TI - The practice of theoretical neuroscience. PMID- 16306886 TI - Neural mechanisms of attention and control: losing our inhibitions? PMID- 16306887 TI - Flipping the switch from electrical to chemical communication. PMID- 16306888 TI - Tinker to Evers to Chance: semaphorin signaling takes teamwork. PMID- 16306890 TI - Odor here, odor there: chemosensation and reproductive function. PMID- 16306891 TI - A natural approach to studying vision. AB - An ultimate goal of systems neuroscience is to understand how sensory stimuli encountered in the natural environment are processed by neural circuits. Achieving this goal requires knowledge of both the characteristics of natural stimuli and the response properties of sensory neurons under natural stimulation. Most of our current notions of sensory processing have come from experiments using simple, parametric stimulus sets. However, a growing number of researchers have begun to question whether this approach alone is sufficient for understanding the real-life sensory tasks performed by the organism. Here, focusing on the early visual pathway, we argue that the use of natural stimuli is vital for advancing our understanding of sensory processing. PMID- 16306892 TI - In praise of artifice. AB - The visual system evolved to process natural images, and the goal of visual neuroscience is to understand the computations it uses to do this. Indeed the goal of any theory of visual function is a model that will predict responses to any stimulus, including natural scenes. It has, however, recently become common to take this fundamental principle one step further: trying to use photographic or cinematographic representations of natural scenes (natural stimuli) as primary probes to explore visual computations. This approach is both challenging and controversial, and we argue that this use of natural images is so fraught with difficulty that it is not useful. Traditional methods for exploring visual computations that use artificial stimuli with carefully selected properties have been and continue to be the most effective tools for visual neuroscience. The proper use of natural stimuli is to test models based on responses to these synthetic stimuli, not to replace them. PMID- 16306893 TI - Analyzing receptive fields, classification images and functional images: challenges with opportunities for synergy. AB - In neurophysiology, psychophysics, optical imaging and functional imaging studies, the investigator seeks a relationship between a high-dimensional variable, such as an image, and a categorical variable, such as the presence or absence of a spike or a behavior. The usual analysis strategy is fundamentally identical across these contexts--it amounts to calculating the average value of the high-dimensional variable for each value of the categorical variable and comparing these results by subtraction. Though intuitive and straightforward, this procedure may be inaccurate or inefficient and may overlook important details. Sophisticated approaches have been developed within these several experimental contexts, but they are rarely applied beyond the context in which they were developed. Recognition of the relationships among these contexts has the potential to accelerate improvements in analytic methods and to increase the amount of information that can be gleaned from experiments. PMID- 16306898 TI - The timing of reperfusion therapy. PMID- 16306899 TI - Atrial fibrillation ablation: a realistic alternative to pharmacologic therapy. PMID- 16306913 TI - Can surgical septal myectomy improve survival in patients with obstructive hypertrophic cardiomyopathy? PMID- 16306914 TI - Does eplerenone benefit postinfarction patients with heart failure and left ventricular systolic dysfunction? PMID- 16306915 TI - Can EuroSCORE accurately predict long-term outcome after cardiac surgery? PMID- 16306916 TI - Single-chamber versus dual-chamber pacing: which is most efficacious in elderly patients with atrioventricular block? PMID- 16306917 TI - Does PCI reduce the occurrence of 'hard' clinical endpoints in patients with stable coronary artery disease? PMID- 16306918 TI - Do n-3 polyunsaturated fatty acids prevent atrial fibrillation after coronary artery bypass grafting surgery? PMID- 16306919 TI - Chronic heart failure: an overview of conventional treatment versus novel approaches. AB - Chronic heart failure (CHF) is a leading cause of hospitalization and is associated with a poor prognosis, although in the past decade substantial progress has been made in understanding the pathophysiology and therapy of CHF with reduced left ventricular (LV) ejection fraction. Use of angiotensin converting-enzyme inhibitors and angiotensin-receptor antagonists either individually or in combination, certain beta-receptor blockers, and judicious use of aldosterone antagonists, has reduced hospital admission rates and mortality from CHF with reduced LV ejection fraction. More clinical trials are needed, however, particularly in patients with CHF and preserved LV ejection fraction. In patients who remain symptomatic despite medical therapy, and who have long QRS intervals (>0.12 s) and markedly reduced LV ejection fraction, the value of cardiac resynchronization therapy with a biventricular pacemaker has now been demonstrated. Yet, morbidity and mortality remain high, indicating a major need for further improvement. Novel therapies include medical management with statins, vasopressin antagonists, erythropoietin, oxypurinol and levosimendan, which improve vascular and myocardial function and reduce fluid overload, in addition to surgical approaches, which reduce LV remodeling. These routes might not, however, suffice in patients with CHF and LV dysfunction. Prevention of apoptosis and particularly regeneration of cardiac muscle would represent a shift of the current paradigm. Stem-cell-based therapies are rapidly evolving, and while basic science is needed to optimize these strategies, medium-sized clinical studies could help to verify the beneficial effects on LV function. In this review, we discuss current treatment methods and new strategies to improve treatment of CHF. PMID- 16306920 TI - Immunomodulation of atherosclerosis with a vaccine. AB - Experimental observations have established that the innate and adaptive immune mechanisms both have roles in the modulation of atherosclerosis. The complex function that the immune system has in the pathophysiology of atherosclerosis is highlighted by the fact that both proatherogenic and atheroprotective effects of immune activation can be demonstrated. An immune response to the protein and lipid components of oxidized LDL cholesterol has been observed in experimental models, and immunization with these antigens has generally reduced atherosclerosis. The findings suggest the tantalizing possibility that an atheroprotective vaccine can be developed. Our laboratories have identified several antigenic epitopes in the human apolipoprotein B100 component of LDL cholesterol. Active immunization with some of these epitopes has reduced atherosclerosis in hyperlipidemic mice. We believe, therefore, that a vaccine based on apolipoprotein B100-related peptide could have a role in reducing atherosclerosis. In this review, we discuss the possible immunologic mechanisms by which vaccines against atherosclerosis might work and the ways in which such treatment might be most effectively administered. PMID- 16306921 TI - Technology Insight: an overview of research in drug-eluting stents. AB - Drug-eluting stents (DESs) have revolutionized interventional cardiology over the past few years to the extent that balloon angioplasty and bare stents did in the 1980s and 1990s. The first DESs became commercially available in Europe in 2002 and in the US in 2003, and it is estimated that up to 80% of patients who undergo stent implantation in the US now receive a DES. Two devices, Cypher sirolimus eluting stents (Cordis Corporation, Miami Lakes, FL) and Taxus paclitaxel-eluting stents (Boston Scientific Corporation, Natick, MN), are currently licensed for sale in both regions. Multiple new devices using different drugs, carriers and stents are currently undergoing clinical trials to establish their efficacy and obtain approval for commercialization. While the remarkable reduction of restenosis has accounted for the success of DESs, concerns remain regarding long term follow-up; published 3-year follow-up results are available for fewer than 200 patients overall. Reports of late stent thrombosis have emerged, particularly in relation to discontinuation of antiplatelet therapy. In patients treated with DESs, long-term administration of at least one antiplatelet agent must be continued following completion of the mandatory dual antiplatelet regimen. In this review, we summarize the findings available for DESs so far, discuss emerging safety and efficacy data, and look at the future directions for these devices. PMID- 16306922 TI - Ambulatory monitoring of aborted sudden cardiac death related to hypertrophic cardiomyopathy. AB - BACKGROUND: A 47-year-old woman with obstructive hypertrophic cardiomyopathy presented with chest pain that had persisted despite treatment with verapamil and alpha-receptor antagonists. The patient had no other significant cardiac symptoms, no history of hypertension, and no familial predisposition to hypertrophic cardiomyopathy or sudden cardiac death. A loud (grade III/VI), dynamic, systolic ejection murmur was noted that could be heard diffusely over the precordium. INVESTIGATIONS: Radionuclide perfusion imaging, coronary angiography, intracoronary Doppler flow measurements, and ambulatory electrocardiographic monitoring. DIAGNOSIS: Obstructive hypertrophic cardiomyopathy, myocardial ischemia and sudden cardiac arrest. MANAGEMENT: Surgical myectomy and cardioverter-defibrillator implantation. PMID- 16306923 TI - Mediterranean diet has no effect on markers of inflammation and metabolic risk factors in patients with coronary artery disease. AB - OBJECTIVE: Mediterranean diet is associated with decreased levels of inflammatory markers and metabolic risk factors in epidemiologic studies and recent trials on patients with metabolic syndrome. Given the recent improvements in medical treatments, it is unclear if such beneficial effects are also present in patients with coronary artery disease (CAD). We therefore investigated the effect of Mediterranean diet on markers of inflammation and metabolic risk factors in patients with treated CAD. DESIGN: Randomized, controlled trial. SUBJECTS: A total of 101 patients (59.4+/-8.6 years, 23% female) with established and treated CAD (80% statins). INTERVENTIONS: Participants were assigned to a Mediterranean diet group (MG; n=48) with a 1-year program of 100 h of education, or to a written advice-only group (AG; n=53). Before and after intervention, we measured serum high-sensitivity C-reactive protein (hs-CRP), fibrinogen, fasting insulin, homocysteine, serum lipids and plasma fatty acids. RESULTS: The Mediterranean diet program increased the intakes of fish, fruits/vegetables and moderately of canola/olive oil and increased plasma concentrations of long-chain n-3 polyunsaturated fatty acids in the MG. Median hs-CRP and mean fibrinogen, homocysteine, fasting insulin, triglycerides and serum cholesterols remained unchanged in both groups. CONCLUSIONS: Adoption of a Mediterranean diet by patients with medically treated CAD has no effect on markers of inflammation and metabolic risk factors. SPONSORSHIP: Alfried Krupp Foundation, Essen, Germany. PMID- 16306924 TI - Urbanization and obesity in The Gambia: a country in the early stages of the demographic transition. AB - OBJECTIVE: To investigate the distribution of overweight and obesity and its relationship with socio-economic and behavioural factors in a developing-country population undergoing rapid nutritional transition. DESIGN: Cross-sectional house to-house survey in urban Gambia. SUBJECTS: Four groups of 50 subjects were sampled as follows: young men (YM, 14-25 years), young women (YW, 14-25 years), older men (OM, 35-50 years) and older women (OW, 35-50 years). MEASUREMENTS: Several socio-economic and behavioural factors were investigated. Composite indices for socio-economic status, education, healthy lifestyle and western influences were created. Body weight, height, waist and hip circumferences were measured and body mass index (BMI) was calculated. Body composition was assessed by leg-to-leg bioimpedance. Overweight was defined as BMI=25.0-29.9 kg/m(2) and obesity as BMI>or=30.0 kg/m(2). RESULTS: There were highly significant gender and age differences in overweight (YM=0%, YW=10%, OM=6% and OW=34%) and obesity (YM=0%, YW=4%, OM=6% and OW=50%). Only 16% of OW were neither overweight nor obese compared to 88% of OM. OW had a higher fat mass percent (38.4%) than other groups, while fat-free mass (kg) was significantly higher in males than females with YW having the lowest value. Young generations were more educated and more influenced by western ideals than OM and OW. Weight gain was not always associated with weight concern and many overweight/obese subjects did not perceive themselves as overweight. CONCLUSION: Social and behavioural changes are already creating a perceptible 'generational gap' among this population undergoing rapid transition. The improved education and current lean status of the younger adults offers opportunities for preventative interventions. These need to be specially targeted at women. PMID- 16306925 TI - Efficacy of combined iron and zinc supplementation on micronutrient status and growth in Vietnamese infants. AB - OBJECTIVE: To evaluate the effect of combined iron-zinc supplementation on micronutrient status, growth and morbidity. DESIGN: Randomized, double-masked, placebo-controlled supplementation trial. SETTING: Rural district of Que Vo, in the Red River Delta in Vietnam. SUBJECTS: A total of 915 breast-fed infants aged 4-7 months were included and 784 completed the study. INTERVENTIONS: The Fe-group received daily and for a 6-month period 10 mg of iron, the Zn-group 10 mg zinc, the Fe-Zn group 10 mg iron+10 mg zinc and the placebo group a placebo. Hemoglobin (Hb), serum ferritin (SF) and zinc (SZn), and anthropometry were measured before and at the end of the intervention. Morbidity was recorded daily. RESULTS: Changes of Hb and SF were higher in both Fe and Fe+Zn groups (respectively 22.6 and 20.6 g/l for Hb; 36.0 and 24.8 microg/l for SF) compared to Zn and placebo groups (Hb: 6.4 and 9.8 g/l; SF: -18.2 and -16.9 microg/l, P<0.0001). SZn increased more in Zn group (10.3 micromol/l) than in Fe+Zn group (8.0 micromol/l, P=0.03) and more in these groups compared to Fe and placebo groups (1.6 and 1.2 micromol/l, P<0.0001). Weight gain was higher in the Zn group. No significant effects of supplementations on growth in length or morbidity. CONCLUSIONS: Combined iron-zinc supplementation had a positive effect on iron and zinc status in infants. However, the positive effect of zinc alone on SZn and weight would indicate a negative interaction of iron when added to zinc supplements. SPONSORSHIP: UNICEF New York. PMID- 16306926 TI - Risk factors for goiter in primary school girls in Qom city of Iran. AB - OBJECTIVE: Goiter is endemic in Iran. The iodine deficiency disorders program was begun a few years ago in Iran, and the coverage of iodized salt is sufficient now. But, in a periodic yearly medical examination of primary school girls in Qom, the prevalence of goiter was above 30% in 2002. This survey was designed to study the risk factors of goiter in those students. DESIGN: The study was a randomized (multistage, proportional simple random sampling) case-control study. SUBJECTS AND INTERVENTIONS: We selected and performed thyroid examinations in 1050 girl students in primary schools in Qom city of Iran in 2002. We found 284 cases: girls in primary schools had goiter in accordance with the clinical exam of World Health Organization classification. Among students who did not present with goiter in the clinical exam, we randomly selected 288 students as the control group. We used a questionnaire to evaluate them for the risk factors of goiter. RESULTS: The mean+/-s.d. ages of cases and controls were 8.7+/-1.3 and 8.9+/-1.3 years, respectively. There is no significant difference between the two groups regarding history of soya, kale, turnip, fish, daily iodized salt usage, education and job of mothers, monthly family income, nationality, immigration and residential situation. By using multinomial logistic regression, we found that storage of iodized salt in open containers, odds ratio (OR): 2.201 (1.412-3.428); P-value <0.0001, medium socioeconomic situation (SES) of family, OR: 2.099 (1.029 4.282), P-value=0.041, district 2 of Qom city, OR: 2.880 (1.376-6.027), P value=0.005, and district 3 of Qom city, OR: 2.051(1.032-4.078), P-value=0.041, were the major risk factors for goiter in this population. CONCLUSIONS: In this study, the main risk factors for goiter were storage of iodized salt in open containers, medium SES and also living in specific districts of Qom city. As the coverage of iodinized salt is over 95% in Iran, we advise the education of the family about storage of iodized salt in closed containers. We also recommend the study of the other risk factors of goiter in the different geographical areas of Iran, because of differences in the SES and nutritional habits. SPONSORSHIP: This study was supported by issuing permission letters for our activities: (not funding support) Qom Health Network and Medical Services, Qom Medical University, Qom Primary School Education Office, Fathemieh Medical University. PMID- 16306927 TI - Validity of a questionnaire to assess fruit and vegetable intake in adults. AB - OBJECTIVE: To validate a method for assessing fruit and vegetable intake among adults in a population of low intake. The method assesses mean intake and ranks individuals by their usual intake. DESIGN: A precoded fruit and vegetable Questionnaire included a 24-h recall and a food frequency. The participants filled in the Questionnaire, a week later they started a 7-day food record, 1-day weighed record and 6 days using household measures. SUBJECTS: Following advertisements 40 participants were recruited, 36 returned food records(mean age=37 years). RESULTS: No difference was observed between the average intake yielded by the 24-h recall and that from the 1-day weighed food record of fruits or vegetables. Correlation coefficients between results from the food frequency questionnaire and the 7-day food records were 0.45 (P=0.007) for vegetables, 0.63 (P<0.001) for fruits and 0.73 (P<0.001) for fruits and vegetables combined. Cross classification into quartiles showed that the proportion of participants in the same or the adjacent quartile of the intake distribution were 94% for fruit intake and 80% for vegetable intake. Registered intake of fruits and vegetables was higher the first 4 days of the record than the last 3 days (P=0.002). The 4 day food record correlated with the food frequency questionnaire in similar manner as the 7-day record. CONCLUSION: The present study indicates that the precoded 24-h recall may be valuable tool for measuring average intake of fruits and vegetables among adults in a population of low intake. Moreover, the food frequency questionnaire was valid for ranking individuals according to their usual intake. A 4-day food record might be sufficient when validating food frequency questionnaires for fruits and vegetables. PMID- 16306928 TI - Low-cost diets: more energy, fewer nutrients. AB - Energy-dense diets offer a low-cost dietary option to the consumer. However, they are more likely to be nutrient-poor. In this study, based on the French national food consumption survey, the diet costs were estimated using retail food prices in France. Adult participants were stratified by quartiles of energy cost (in euro/10 MJ). Dietary energy density, energy and nutrient intakes were then compared across groups. Participants in the lowest quartile of energy cost had the highest energy intakes, the most energy-dense diets and the lowest daily intakes of key vitamins and micronutrients. Participants in the highest quartile of energy cost had lower energy intakes, and diets that were higher in nutrients and lower in energy density. However, their daily diet costs were 165% higher. In this observational study, the more nutrient-dense diets were associated with higher diet costs. PMID- 16306929 TI - Breast and complementary feeding practices in relation to morbidity and growth in Malawian infants. AB - OBJECTIVES: The objective of this study was to compare growth, morbidity incidence and risk factors for undernutrition between infants receiving complementary feeding early, before 3 months of age, with those receiving complementary foods after 3 months in a poor rural Malawian community. METHODS: A cohort of babies was enrolled at birth for follow-up to 12 months of age. Weight, length, morbidity and feeding patterns were recorded at 4 weekly intervals from birth to 52 weeks. RESULTS: Mean age at introduction of water was 2.5 months (range 0-11.8), complementary foods 3.4 months (range, 1.0-10.7) and solids 4.5 months (range 1.2-13.8). Over 40% of infants had received complementary foods by 2 months and 65% by 3 months. The proportion of exclusively breast-fed infants, which included those receiving supplemental water, was 13% at 4 months, 6.3% at 5 months and 1.5% at 6 months. Infants with early complementary feeding had lower weight for age at 3 and 6 months (P<0.05), and at 9 months (P=0.07) and at 2 months they were approximately 200 g lighter. Early complementary feeding was significantly associated with increased risk for respiratory infection (P<0.05), and marginally increased risk for eye infection and episodes of malaria. Maternal illiteracy was associated with early complementary feeding (OR=2.1, 95% CI 1.3, 3.2), while later complementary feeding was associated with reduced infant morbidity and improved growth. CONCLUSION: Breast-feeding promotion programmes should target illiterate women. Greater emphasis is required to improve complementary feeding practices. PMID- 16306930 TI - Significant changes in physical activity among pregnant women in the UK as assessed by accelerometry and self-reported activity. AB - OBJECTIVE: Research on the impact of maternal physical activity on pregnancy outcomes has often employed subjective measures of physical activity obtained by diary or questionnaire. This study investigates the feasibility of using accelerometry as an objective measure of physical activity of pregnant women compared with subjective data obtained via activity recall among pregnant women. DESIGN: Activity data were collected prospectively on 57 women at 12, 16, 25, 34 and 38 weeks of gestation. Total daily physical activity was assessed by ambulatory accelerometer and activity interview (self-report). Maternal personality variables (health value, extroversion) were assessed by established scales. SETTING: Leicestershire, UK. SUBJECTS: Pregnant women were recruited by voluntary participation via antenatal booking clinics. In all, 64 pregnant women with low-risk pregnancy were enrolled onto the study, of whom 57 completed the study. RESULTS: Mean 24 h physical activity levels (PAL) decreased significantly from second to third trimester as assessed by self-report interview (1.51-1.29 Metabolic Equivalent TEE-h/day, P<0.01) and accelerometry (200.05-147.42 counts/min, P<0.01). The correlation between the two measures declined as pregnancy progressed (r value ranging from 0.55 to 0.08). Compliance with the accelerometers declined from 90% at 12 weeks to 47% at 34 weeks (P<0.01). Compliance with the self-report interviews was 100%. Those who fully complied with the accelerometry demonstrated a significantly higher health value (P<0.05) and a significantly greater level of extroversion (P<0.05) than those who did not. CONCLUSIONS: Accelerometers and self-reported activity interviews both indicated a significant decline in PAL during pregnancy. Although subjects showed a willingness to use both methods, accelerometers resulted in variable compliance with 72 h monitoring. Both techniques may be limited by the need to measure low levels of physical activity during the third trimester. SPONSORSHIP: Cambridge Neurotechnology Ltd, UK, assisted with the provision of Actiwatch accelerometers. PMID- 16306931 TI - Follow-up of diet and cardiovascular risk factors 20 years after cessation of intervention in the Oslo Diet and Antismoking Study. AB - OBJECTIVE: The Oslo Diet and Antismoking study was a 5-year randomised trial initiated in 1972-1973, which studied the effect of dietary change and smoking cessation for the prevention of coronary heart disease among high-risk middle aged men. To test the long-term maintenance of lifestyle change, we examined diet and cardiovascular risk factors in subjects initially randomised to the control and intervention groups 20 years after cessation of the intervention. SUBJECTS AND DESIGN: Of the original cohort that included 1232 participants, 910 survivors were identified in 1997 and cardiovascular risk factors were measured in 563 (62%) in 1997-1999. Of these, 558 (99%) also completed questionnaires about their food intake and attitudes to health and diet. RESULTS: Cigarette smoking was nearly halved between baseline and 20-year follow-up in each of the intervention and control groups (P<0.001 within groups), but did not differ between the intervention group (39%) versus the control group (34%); P=0.07. Body mass index increased by 1.4+/-2.6 and 1.6+/-2.6 kg/m(2) between baseline and 20-year follow up in the intervention and control groups, respectively (P<0.001 within groups; NS between groups). Serum total cholesterol and triglyceride concentrations decreased substantially in subjects treated or untreated with statins (P<0.001 within the intervention and control groups) but did not differ between the groups (total cholesterol change of -1.4+/-1.3 and -1.3+/-1.2 mmol/l, respectively, and triglyceride change of -0.5+/-1.0 mmol/l in both groups). Men in the intervention group reported a less atherogenic fat quality score and lower intakes of fat, saturated fat and cholesterol, higher intakes of long chain polyunsaturated fatty acids, protein and beta-carotene and greater attention to lifestyle and change of diet than the control group (all P<0.05). The fatty acid concentrations did not differ, however, between the intervention and control groups (P>0.05). CONCLUSIONS: No long-term differences in smoking rates or lipid concentrations between the intervention and control groups were observed in the surviving attendees two decades after the end of the trial. Lifestyle intervention still influenced the dietary intake, though modestly. PMID- 16306932 TI - Salty taste acceptance by infants and young children is related to birth weight: longitudinal analysis of infants within the normal birth weight range. AB - BACKGROUND: Birth weight and sodium intake are both associated with risk for hypertension. It is not known whether birth weight influences response to salty taste. OBJECTIVE: To assess the relationship between birth weight and salty taste acceptance of infants and young children. DESIGN: Acceptance of salty taste was assessed at 2 (n = 80) and 6 (n = 76) months in infants (birth weight >2.5 kg) enrolled in a prospective cohort study. Acceptance was expressed as proportional intake following 1-min ingestion tests with water and salt solutions (0.17 and 0.34 mol/l NaCl, in water). Birth weight was obtained by maternal report. Questionnaires completed by mothers and food-ranking procedures performed by children evaluated salt liking and preference in a subset (n = 38) of subjects at preschool age (36 or 48 months). SETTING: Nonprofit basic research institute in Philadelphia, PA, USA. RESULTS: Regression analysis revealed significant negative associations between birth weight and acceptance of salty taste at 2 months (0.17 mol/l, P < 0.0001; 0.34 mol/l, P < 0.01) but not at 6 months. Relationships were not affected by adjustment for potential confounders. In preschoolers, greater liking of (P < 0.05) and preference for (P < 0.01) salty foods was associated with lower birth weight in simple, but not adjusted, models. CONCLUSION: Measures related to salty taste preference were inversely related to birth weight over the first 4 years of life. Additional studies should substantiate these findings and explore whether early response to salty taste predicts future sodium intake, blood pressure, or other public health-related outcomes. SPONSORSHIP: National Institutes of Health (DC 00882). PMID- 16306933 TI - Effect of long-term mild hypothermia or short-term mild hypothermia on outcome of patients with severe traumatic brain injury. AB - To compare the effect of long-term mild hypothermia versus short-term mild hypothermia on the outcome of 215 severe traumatic brain injured patients with cerebral contusion and intracranial hypertension. At three medical centers, 215 patients aged 18 to 45 years old with an admission Glasgow Coma Scale < or =8 within 4 h after injury were randomly divided into two groups: long-term mild hypothermia group (n = 108) for 5+/-1.3 days mild hypothermia therapy and short term mild hypothermia group (n = 107) for 2+/-0.6 days mild hypothermia therapy. All patients had intracranial hypertension and frontotemporoparietal contusion with midline shift >1 cm confirmed on computed tomographic scan. Glasgow Outcome Scale at 6-month follow-up, 47 cases had favorable outcome (43.5%), and other 61 cases had unfavorable outcome (56.5%) in the long-term mild hypothermia group. However, only 31 cases had favorable outcome (29.0%), and other 76 cases had unfavorable outcome (71.0%) in the short-term mild hypothermia group (P < 0.05). The intracranial pressure significantly rebounded after rewarming in the short term mild hypothermia group, but not in the long-term mild hypothermia (P < 0.05). Furthermore, the incidence of stress ulcer, epilepsy, pulmonary infection, intracranial infection did not significantly differ between the two groups (P > 0.05). Compared with short-term mild hypothermia, long-term mild hypothermia significantly improves the outcome of severe traumatic brain injured patients with cerebral contusion and intracranial hypertension without significant complications. Our data suggest that 5 days of long-term cooling is more efficacious than 2 days of short-term cooling when mild hypothermia is used to control refractory intracranial hypertension in patients with severe traumatic brain injury. PMID- 16306934 TI - The rat blood-brain barrier transcriptome. AB - The blood-brain barrier (BBB) is the cellular interface between the circulating blood and neural environment, and is created by apposed endothelial cells and their intercellular tight junctions. Many aspects of how the BBB functions at the molecular level remain unresolved; therefore, we report for the first time a comprehensive gene expression profile of rat brain microvessels using serial analysis of gene expression (SAGE). We assembled a full and quantitative SAGE catalog containing 101,364 tags, of which 33% of the tags matched known genes, 51% matched expressed sequence tags (ESTs) in the Unigene database, and 16% of the tags were unassigned. The transcriptome catalog contains many new and novel transcripts among known BBB genes. A large compliment of junctional proteins and an extensive assortment of facilitated carrier and ATP-dependent transporters are included. To identify microvessel-enriched transcripts, we compared the microvessel SAGE catalog to cortex and hippocampus SAGE catalogs. This resulted in identification of 864 genes, including several known for their abundant expression at the BBB, such as the transferrin receptor (TrnR). Sorting enriched genes based on function revealed groups that encode transporters (11%), receptors (5%), proteins involved in vesicle trafficking (4%), structural proteins (10%), and components of signal transduction pathways (17%). This genomic repertoire emphasizes the unique cellular phenotype existing within the brain and further implicates the BBB as a mediator between the brain and periphery. These results may provide a useful resource and reference point from which to determine the effects of different physiological, developmental, and disease processes on BBB gene expression. PMID- 16306935 TI - Effect of duration of osmotherapy on blood-brain barrier disruption and regional cerebral edema after experimental stroke. AB - Osmotherapy is the cornerstone of medical management for cerebral edema associated with large ischemic strokes. We determined the effect of duration of graded increases in serum osmolality with mannitol and hypertonic saline (HS) on blood-brain barrier (BBB) disruption and regional cerebral edema in a well characterized rat model of large ischemic stroke. Halothane-anesthetized adult male Wistar rats were subjected to transient (2-h) middle cerebral artery occlusion (MCAO) by the intraluminal occlusion technique. Beginning at 6 h after MCAO, rats were treated with either no intravenous fluids or a continuous intravenous infusion (0.3 mL/h) of 0.9% saline, 20% mannitol, 3% HS, or 7.5% HS for 24, 48, 72, and 96 h. In the first series of experiments, BBB permeability was quantified by the Evans blue (EB) extravasation method. In the second series of experiments, water content was assessed by comparing wet-to-dry weight ratios in six predetermined brain regions. Blood-brain barrier disruption was maximal in rats treated with 0.9% saline for 48 h, but did not correlate with increases in serum osmolality or treatment duration with osmotic agents. Treatment with 7.5% HS attenuated water content in the periinfarct regions and all subregions of the contralateral nonischemic hemisphere to a greater extent than mannitol did with no adverse effect on survival rates. These data show that (1) BBB integrity is not affected by the duration and degree of serum osmolality with osmotic agents, and (2) attenuation of increases in brain water content with HS to target levels >350 mOsm/L may have therapeutic implications in the treatment of cerebral edema associated with ischemic stroke. PMID- 16306936 TI - Critical role of TRAF3 in the Toll-like receptor-dependent and -independent antiviral response. AB - Type I interferon (IFN) production is a critical component of the innate defence against viral infections. Viral products induce strong type I IFN responses through the activation of Toll-like receptors (TLRs) and intracellular cytoplasmic receptors such as protein kinase R (PKR). Here we demonstrate that cells lacking TRAF3, a member of the TNF receptor-associated factor family, are defective in type I IFN responses activated by several different TLRs. Furthermore, we show that TRAF3 associates with the TLR adaptors TRIF and IRAK1, as well as downstream IRF3/7 kinases TBK1 and IKK-epsilon, suggesting that TRAF3 serves as a critical link between TLR adaptors and downstream regulatory kinases important for IRF activation. In addition to TLR stimulation, we also show that TRAF3-deficient fibroblasts are defective in their type I IFN response to direct infection with vesicular stomatitis virus, indicating that TRAF3 is also an important component of TLR-independent viral recognition pathways. Our data demonstrate that TRAF3 is a major regulator of type I IFN production and the innate antiviral response. PMID- 16306937 TI - Specificity in Toll-like receptor signalling through distinct effector functions of TRAF3 and TRAF6. AB - Toll-like receptors (TLRs) are activated by pathogen-associated molecular patterns to induce innate immune responses and production of pro-inflammatory cytokines, interferons and anti-inflammatory cytokines. TLRs activate downstream effectors through adaptors that contain Toll/interleukin-1 receptor (TIR) domains, but the mechanisms accounting for diversification of TLR effector functions are unclear. To dissect biochemically TLR signalling, we established a system for isolating signalling complexes assembled by dimerized adaptors. Using MyD88 as a prototypical adaptor, we identified TNF receptor-associated factor 3 (TRAF3) as a new component of TIR signalling complexes that is recruited along with TRAF6. Using myeloid cells from TRAF3- and TRAF6-deficient mice, we show that TRAF3 is essential for the induction of type I interferons (IFN) and the anti-inflammatory cytokine interleukin-10 (IL-10), but is dispensable for expression of pro-inflammatory cytokines. In fact, TRAF3-deficient cells overproduce pro-inflammatory cytokines owing to defective IL-10 production. Despite their structural similarity, the functions of TRAF3 and TRAF6 are largely distinct. TRAF3 is also recruited to the adaptor TRIF (Toll/IL-1 receptor domain containing adaptor-inducing IFN-beta) and is required for marshalling the protein kinase TBK1 (also called NAK) into TIR signalling complexes, thereby explaining its unique role in activation of the IFN response. PMID- 16306938 TI - An siRNA-based microbicide protects mice from lethal herpes simplex virus 2 infection. AB - Herpes simplex virus 2 (HSV-2) infection causes significant morbidity and is an important cofactor for the transmission of HIV infection. A microbicide to prevent sexual transmission of HSV-2 would contribute substantially to controlling the spread of HIV and other infections. Because RNA interference (RNAi) provides effective antiviral defence in plants and other organisms, several studies have focused on harnessing RNAi to inhibit viral infection. Here we show that vaginal instillation of small interfering RNAs (siRNAs) targeting HSV-2 protects mice from lethal infection. siRNAs mixed with lipid are efficiently taken up by epithelial and lamina propria cells and silence gene expression in the mouse vagina and ectocervix for at least nine days. Intravaginal application of siRNAs targeting the HSV-2 UL27 and UL29 genes (which encode an envelope glycoprotein and a DNA binding protein, respectively) was well tolerated, did not induce interferon-responsive genes or cause inflammation, and protected mice when administered before and/or after lethal HSV-2 challenge. These results suggest that siRNAs are attractive candidates for the active component of a microbicide designed to prevent viral infection or transmission. PMID- 16306940 TI - Immunology: Insulin auto-antigenicity in type 1 diabetes. AB - Spontaneous type 1 diabetes occurs when the autoimmune destruction of pancreatic beta-islet cells prevents production of the hormone insulin. This causes an inability to regulate glucose metabolism, which results in dangerously raised blood glucose concentrations. It is generally accepted that thymus-derived lymphocytes (T cells) are critically involved in the onset and progression of type 1 diabetes, but the antigens that initiate and drive this destructive process remain poorly characterized--although several candidates have been considered. Nakayama et al. and Kent et al. claim that insulin itself is the primary autoantigen that initiates spontaneous type 1 diabetes in mice and humans, respectively, a result that could have implications for more effective prevention and therapy. However, I believe that this proposed immunological role of insulin may be undermined by the atypical responses of T cells to the human insulin fragment that are described by Kent et al.. PMID- 16306945 TI - Google makes data free for all. PMID- 16306946 TI - US watchdog finds bias against morning-after pill. PMID- 16306947 TI - Seals net data from cold seas. PMID- 16306948 TI - Neuroscientists put gene therapy into reverse. PMID- 16306950 TI - Korean stem-cell crisis deepens. PMID- 16306951 TI - China steps up drive to vaccinate all domestic birds. PMID- 16306952 TI - Deal on toxicity law fails to appease. PMID- 16306953 TI - Software shakes up schizophrenia diagnosis. PMID- 16306956 TI - Energy in California: power struggle. PMID- 16306957 TI - Antarctic astronomy: seeing in the dark. PMID- 16306958 TI - Synthetic biology: designs on life. PMID- 16306959 TI - Property rights go East. PMID- 16306962 TI - Librarians can help prevent accidental plagiarism. PMID- 16306963 TI - Is the ID debate proof of an intelligent deceiver? PMID- 16306964 TI - Evidence of group learning does not add up to culture. PMID- 16306965 TI - Bush's policy stopped US gaining stem-cell lead. PMID- 16306966 TI - Let us go forth and safely multiply. PMID- 16306972 TI - Pushing for power. PMID- 16306973 TI - Quantum computing: a bit chilly. PMID- 16306974 TI - Cell biology: silenced RNA on the move. PMID- 16306976 TI - Condensed-matter physics: focus on the Fermi surface. PMID- 16306977 TI - Obesity: aquaporin enters the picture. PMID- 16306978 TI - Photonics: Winged light. PMID- 16306979 TI - Histocompatibility: colonial match and mismatch. PMID- 16306980 TI - Synthetic biology: engineering Escherichia coli to see light. AB - We have designed a bacterial system that is switched between different states by red light. The system consists of a synthetic sensor kinase that allows a lawn of bacteria to function as a biological film, such that the projection of a pattern of light on to the bacteria produces a high-definition (about 100 megapixels per square inch), two-dimensional chemical image. This spatial control of bacterial gene expression could be used to 'print' complex biological materials, for example, and to investigate signalling pathways through precise spatial and temporal control of their phosphorylation steps. PMID- 16306981 TI - Insect communication: 'no entry' signal in ant foraging. AB - Forager ants lay attractive trail pheromones to guide nestmates to food, but the effectiveness of foraging networks might be improved if pheromones could also be used to repel foragers from unrewarding routes. Here we present empirical evidence for such a negative trail pheromone, deployed by Pharaoh's ants (Monomorium pharaonis) as a 'no entry' signal to mark an unrewarding foraging path. This finding constitutes another example of the sophisticated control mechanisms used in self-organized ant colonies. PMID- 16306982 TI - Reconstruction of genetic circuits. AB - The complex genetic circuits found in cells are ordinarily studied by analysis of genetic and biochemical perturbations. The inherent modularity of biological components like genes and proteins enables a complementary approach: one can construct and analyse synthetic genetic circuits based on their natural counterparts. Such synthetic circuits can be used as simple in vivo models to explore the relation between the structure and function of a genetic circuit. Here we describe recent progress in this area of synthetic biology, highlighting newly developed genetic components and biological lessons learned from this approach. PMID- 16306983 TI - Foundations for engineering biology. AB - Engineered biological systems have been used to manipulate information, construct materials, process chemicals, produce energy, provide food, and help maintain or enhance human health and our environment. Unfortunately, our ability to quickly and reliably engineer biological systems that behave as expected remains quite limited. Foundational technologies that make routine the engineering of biology are needed. Vibrant, open research communities and strategic leadership are necessary to ensure that the development and application of biological technologies remains overwhelmingly constructive. PMID- 16306984 TI - Isolation and characterization of a protochordate histocompatibility locus. AB - Histocompatibility--the ability of an organism to distinguish its own cells and tissue from those of another--is a universal phenomenon in the Metazoa. In vertebrates, histocompatibility is a function of the immune system controlled by a highly polymorphic major histocompatibility complex (MHC), which encodes proteins that target foreign molecules for immune cell recognition. The association of the MHC and immune function suggests an evolutionary relationship between metazoan histocompatibility and the origins of vertebrate immunity. However, the MHC of vertebrates is the only functionally characterized histocompatibility system; the mechanisms underlying this process in non vertebrates are unknown. A primitive chordate, the ascidian Botryllus schlosseri, also undergoes a histocompatibility reaction controlled by a highly polymorphic locus. Here we describe the isolation of a candidate gene encoding an immunoglobulin superfamily member that, by itself, predicts the outcome of histocompatibility reactions. This is the first non-vertebrate histocompatibility gene described, and may provide insights into the evolution of vertebrate adaptive immunity. PMID- 16306985 TI - Direct detection of a magnetic field in the innermost regions of an accretion disk. AB - Models predict that magnetic fields play a crucial role in the physics of astrophysical accretion disks and their associated winds and jets. For example, the rotation of the disk twists around the rotation axis the initially vertical magnetic field, which responds by slowing down the plasma in the disk and by causing it to fall towards the central star. The magnetic energy flux produced in this process points away from the disk, pushing the surface plasma outwards, leading to a wind from the disk and sometimes a collimated jet. But these predictions have hitherto not been supported by observations. Here we report the direct detection of the magnetic field in the core of the protostellar accretion disk FU Orionis. The surface field reaches strengths of about 1 kG close to the centre of the disk, and it includes a significant azimuthal component, in good agreement with recent models. But we find that the field is very filamentary and slows down the disk plasma much more than models predict, which may explain why FU Ori fails to collimate its wind into a jet. PMID- 16306986 TI - Experimental implementation of heat-bath algorithmic cooling using solid-state nuclear magnetic resonance. AB - The counter-intuitive properties of quantum mechanics have the potential to revolutionize information processing by enabling the development of efficient algorithms with no known classical counterparts. Harnessing this power requires the development of a set of building blocks, one of which is a method to initialize the set of quantum bits (qubits) to a known state. Additionally, fresh ancillary qubits must be available during the course of computation to achieve fault tolerance. In any physical system used to implement quantum computation, one must therefore be able to selectively and dynamically remove entropy from the part of the system that is to be mapped to qubits. One such method is an 'open system' cooling protocol in which a subset of qubits can be brought into contact with an external system of large heat capacity. Theoretical efforts have led to an implementation-independent cooling procedure, namely heat-bath algorithmic cooling. These efforts have culminated with the proposal of an optimal algorithm, the partner-pairing algorithm, which was used to compute the physical limits of heat-bath algorithmic cooling. Here we report the experimental realization of multi-step cooling of a quantum system via heat-bath algorithmic cooling. The experiment was carried out using nuclear magnetic resonance of a solid-state ensemble three-qubit system. We demonstrate the repeated repolarization of a particular qubit to an effective spin-bath temperature, and alternating logical operations within the three-qubit subspace to ultimately cool a second qubit below this temperature. Demonstration of the control necessary for these operations represents an important step forward in the manipulation of solid state nuclear magnetic resonance qubits. PMID- 16306987 TI - Nodal quasiparticle in pseudogapped colossal magnetoresistive manganites. AB - A characteristic feature of the copper oxide high-temperature superconductors is the dichotomy between the electronic excitations along the nodal (diagonal) and antinodal (parallel to the Cu-O bonds) directions in momentum space, generally assumed to be linked to the 'd-wave' symmetry of the superconducting state. Angle resolved photoemission measurements in the superconducting state have revealed a quasiparticle spectrum with a d-wave gap structure that exhibits a maximum along the antinodal direction and vanishes along the nodal direction. Subsequent measurements have shown that, at low doping levels, this gap structure persists even in the high-temperature metallic state, although the nodal points of the superconducting state spread out in finite 'Fermi arcs'. This is the so-called pseudogap phase, and it has been assumed that it is closely linked to the superconducting state, either by assigning it to fluctuating superconductivity or by invoking orders which are natural competitors of d-wave superconductors. Here we report experimental evidence that a very similar pseudogap state with a nodal antinodal dichotomous character exists in a system that is markedly different from a superconductor: the ferromagnetic metallic groundstate of the colossal magnetoresistive bilayer manganite La1.2Sr1.8Mn2O7. Our findings therefore cast doubt on the assumption that the pseudogap state in the copper oxides and the nodal-antinodal dichotomy are hallmarks of the superconductivity state. PMID- 16306988 TI - Probing carrier dynamics in nanostructures by picosecond cathodoluminescence. AB - Picosecond and femtosecond spectroscopy allow the detailed study of carrier dynamics in nanostructured materials. In such experiments, a laser pulse normally excites several nanostructures at once. However, spectroscopic information may also be acquired using pulses from an electron beam in a modern electron microscope, exploiting a phenomenon called cathodoluminescence. This approach offers several advantages. The multimode imaging capabilities of the electron microscope enable the correlation of optical properties (via cathodoluminescence) with surface morphology (secondary electron mode) at the nanometre scale. The broad energy range of the electrons can excite wide-bandgap materials, such as diamond- or gallium-nitride-based structures that are not easily excited by conventional optical means. But perhaps most intriguingly, the small beam can probe a single selected nanostructure. Here we apply an original time-resolved cathodoluminescence set-up to describe carrier dynamics within single gallium arsenide-based pyramidal nanostructures with a time resolution of 10 picoseconds and a spatial resolution of 50 nanometres. The behaviour of such charge carriers could be useful for evaluating elementary components in quantum computers, optical quantum gates or single photon sources for quantum cryptography. PMID- 16306989 TI - Impacts of orbital forcing and atmospheric carbon dioxide on Miocene ice-sheet expansion. AB - The processes causing the middle Miocene global cooling, which marked the Earth's final transition into an 'icehouse' climate about 13.9 million years ago (Myr ago), remain enigmatic. Tectonically driven circulation changes and variations in atmospheric carbon dioxide levels have been suggested as driving mechanisms, but the lack of adequately preserved sedimentary successions has made rigorous testing of these hypotheses difficult. Here we present high-resolution climate proxy records, covering the period from 14.7 to 12.7 million years ago, from two complete sediment cores from the northwest and southeast subtropical Pacific Ocean. Using new chronologies through the correlation to the latest orbital model, we find relatively constant, low summer insolation over Antarctica coincident with declining atmospheric carbon dioxide levels at the time of Antarctic ice-sheet expansion and global cooling, suggesting a causal link. We surmise that the thermal isolation of Antarctica played a role in providing sustained long-term climatic boundary conditions propitious for ice-sheet formation. Our data document that Antarctic glaciation was rapid, taking place within two obliquity cycles, and coincided with a striking transition from obliquity to eccentricity as the drivers of climatic change. PMID- 16306990 TI - Density of hydrous silicate melt at the conditions of Earth's deep upper mantle. AB - The chemical evolution of the Earth and the terrestrial planets is largely controlled by the density of silicate melts. If melt density is higher than that of the surrounding solid, incompatible elements dissolved in the melt will be sequestered in the deep mantle. Previous studies on dry (water-free) melts showed that the density of silicate melts can be higher than that of surrounding solids under deep mantle conditions. However, melts formed under deep mantle conditions are also likely to contain some water, which will reduce the melt density. Here we present data constraining the density of hydrous silicate melt at the conditions of approximately 410 km depth. We show that the water in the silicate melt is more compressible than the other components, and therefore the effect of water in reducing melt density is markedly diminished under high-pressure conditions. Our study indicates that there is a range of conditions under which a (hydrous) melt could be trapped at the 410-km boundary and hence incompatible elements could be sequestered in the deep mantle, although these conditions are sensitive to melt composition as well as the composition of the surrounding mantle. PMID- 16306991 TI - The entomological inoculation rate and Plasmodium falciparum infection in African children. AB - Malaria is an important cause of global morbidity and mortality. The fact that some people are bitten more often than others has a large effect on the relationship between risk factors and prevalence of vector-borne diseases. Here we develop a mathematical framework that allows us to estimate the heterogeneity of infection rates from the relationship between rates of infectious bites and community prevalence. We apply this framework to a large, published data set that combines malaria measurements from more than 90 communities. We find strong evidence that heterogeneous biting or heterogeneous susceptibility to infection are important and pervasive factors determining the prevalence of infection: 20% of people receive 80% of all infections. We also find that individual infections last about six months on average, per infectious bite, and children who clear infections are not immune to new infections. The results have important implications for public health interventions: the success of malaria control will depend heavily on whether efforts are targeted at those who are most at risk of infection. PMID- 16306992 TI - Neural measures reveal individual differences in controlling access to working memory. AB - The capacity of visual short-term memory is highly limited, maintaining only three to four objects simultaneously. This extreme limitation necessitates efficient mechanisms to select only the most relevant objects from the immediate environment to be represented in memory and to restrict irrelevant items from consuming capacity. Here we report a neurophysiological measure of this memory selection mechanism in humans that gauges an individual's efficiency at excluding irrelevant items from being stored in memory. By examining the moment-by-moment contents of visual memory, we observe that selection efficiency varies substantially across individuals and is strongly predicted by the particular memory capacity of each person. Specifically, high capacity individuals are much more efficient at representing only the relevant items than are low capacity individuals, who inefficiently encode and maintain information about the irrelevant items present in the display. These results provide evidence that under many circumstances low capacity individuals may actually store more information in memory than high capacity individuals. Indeed, this ancillary allocation of memory capacity to irrelevant objects may be a primary source of putative differences in overall storage capacity. PMID- 16306993 TI - Design principles of a bacterial signalling network. AB - Cellular biochemical networks have to function in a noisy environment using imperfect components. In particular, networks involved in gene regulation or signal transduction allow only for small output tolerances, and the underlying network structures can be expected to have undergone evolution for inherent robustness against perturbations. Here we combine theoretical and experimental analyses to investigate an optimal design for the signalling network of bacterial chemotaxis, one of the most thoroughly studied signalling networks in biology. We experimentally determine the extent of intercellular variations in the expression levels of chemotaxis proteins and use computer simulations to quantify the robustness of several hypothetical chemotaxis pathway topologies to such gene expression noise. We demonstrate that among these topologies the experimentally established chemotaxis network of Escherichia coli has the smallest sufficiently robust network structure, allowing accurate chemotactic response for almost all individuals within a population. Our results suggest that this pathway has evolved to show an optimal chemotactic performance while minimizing the cost of resources associated with high levels of protein expression. Moreover, the underlying topological design principles compensating for intercellular variations seem to be highly conserved among bacterial chemosensory systems. PMID- 16306994 TI - Spatial regulation of beta-actin translation by Src-dependent phosphorylation of ZBP1. AB - Localization of beta-actin messenger RNA to sites of active actin polymerization modulates cell migration during embryogenesis, differentiation and possibly carcinogenesis. This localization requires the oncofetal protein ZBP1 (Zipcode binding protein 1), which binds to a conserved 54-nucleotide element in the 3' untranslated region of the beta-actin mRNA known as the 'zipcode'. ZBP1 promotes translocation of the beta-actin transcript to actin-rich protrusions in primary fibroblasts and neurons. It is not known how the ZBP1-RNA complex achieves asymmetric protein sorting by localizing beta-actin mRNA. Here we show that chicken ZBP1 modulates the translation of beta-actin mRNA. ZBP1 associates with the beta-actin transcript in the nucleus and prevents premature translation in the cytoplasm by blocking translation initiation. Translation only occurs when the ZBP1-RNA complex reaches its destination at the periphery of the cell. At the endpoint of mRNA transport, the protein kinase Src promotes translation by phosphorylating a key tyrosine residue in ZBP1 that is required for binding to RNA. These sequential events provide both temporal and spatial control over beta actin mRNA translation, which is important for cell migration and neurite outgrowth. PMID- 16306995 TI - Structures of ParB bound to DNA reveal mechanism of partition complex formation. AB - The faithful inheritance of genetic information, which is essential for all organisms, requires accurate DNA partition (segregation) at cell division. In prokaryotes, partition is mediated by par systems, for which the P1 plasmid system of Escherichia coli is a prototype comprising a partition site and two proteins, ParA and ParB. To form the partition complex necessary for segregation, P1 ParB must recognize a complicated arrangement of A-box and B-box DNA motifs located on opposite ends of a sharply bent parS partition site of approximately 74 bp (refs 3-7). Here we describe structures of ParB bound to partition sites. ParB forms an asymmetric dimer with extended amino-terminal HTH (helix-turn helix) domains that contact A-boxes. The two HTH domains emanate from a dimerized DNA-binding module composed of a six-stranded beta-sheet coiled-coil that binds B boxes. Strikingly, these individual DNA-binding modules rotate freely about a flexible linker, enabling them to contact several arrangements of A- and B-boxes. Most notably, each DNA-binding element binds to and thus bridges adjacent DNA duplexes. These unique structural features of ParB explain how this protein can bind complex arrays of A- and B-box elements on adjacent DNA arms of the looped partition site. PMID- 16306996 TI - An induced-fit mechanism to promote peptide bond formation and exclude hydrolysis of peptidyl-tRNA. AB - The large ribosomal subunit catalyses the reaction between the alpha-amino group of the aminoacyl-tRNA bound to the A site and the ester carbon of the peptidyl tRNA bound to the P site, while preventing the nucleophilic attack of water on the ester, which would lead to unprogrammed deacylation of the peptidyl-tRNA. Here we describe three new structures of the large ribosomal subunit of Haloarcula marismortui (Hma) complexed with peptidyl transferase substrate analogues that reveal an induced-fit mechanism in which substrates and active site residues reposition to allow the peptidyl transferase reaction. Proper binding of an aminoacyl-tRNA analogue to the A site induces specific movements of 23S rRNA nucleotides 2618-2620 (Escherichia coli numbering 2583-2585) and 2541(2506), thereby reorienting the ester group of the peptidyl-tRNA and making it accessible for attack. In the absence of the appropriate A-site substrate, the peptidyl transferase centre positions the ester link of the peptidyl-tRNA in a conformation that precludes the catalysed nucleophilic attack by water. Protein release factors may also function, in part, by inducing an active-site rearrangement similar to that produced by the A-site aminoacyl-tRNA, allowing the carbonyl group and water to be positioned for hydrolysis. PMID- 16306998 TI - Effect of telmisartan/hydrochlorothiazide vs lisinopril/hydrochlorothiazide combination on ambulatory blood pressure and cognitive function in elderly hypertensive patients. AB - The aim of this study was to compare the effects of telmisartan/hydrochlorothiazide (HCTZ) vs lisinopril/HCTZ combination on ambulatory blood pressure and cognitive function in elderly hypertensive patients. A total of 160 patients, 76 men and 84 women, aged 61-75 years, with sitting diastolic blood pressure (DBP)>90 mmHg and <110 mmHg and systolic blood pressure (SBP)>140 mmHg were randomized to receive temisartan 80 mg/HCTZ 12.5 mg o.d. or lisinopril 20 mg/HCTZ 12.5 mg o.d. for 24 weeks, according to a prospective, open-label, blinded end point, parallel-group design. At the end of a 2-week wash-out period and after 12 and 24 weeks of active treatment, 24-h noninvasive ambulatory BP monitoring (ABPM) was performed and cognitive function was evaluated through six different tests (verbal fluency, Boston naming test, word-list memory, word-list recall, word-list recognition and Trails B). Both treatments significantly reduced ambulatory BP. However, the telmisartan/HCTZ combination produced a greater reduction in 24-h, day-time and night time ABPM values. Lisinopril/HCTZ did not induce significant changes in any of the cognitive function test scores at any time of the study, whereas at both 12 and 24 weeks telmisartan/HCTZ significantly improved the word-list memory score (+17.1 and +15.7%, respectively, P<0.05 vs baseline), the word-list recall score (+13.5 and +16.9%, P<0.05) and the Trails B score (-33 and -30.5%, P<0.05). These results suggest that in elderly hypertensive patients treatment with telmisartan/HCTZ produces a slightly greater reduction in ambulatory BP than lisinopril/HCTZ combination and, unlike this latter, improves some of the components of cognitive function, particularly episodic memory and visuospatial abilities. PMID- 16306999 TI - Increased arterial stiffness in healthy young South Asian men. PMID- 16307000 TI - Restoration of all dystrophin protein interactions by functional domains in trans does not rescue dystrophy. AB - Rescue of dystrophic skeletal muscle in mdx and utrophin/dystrophin-deficient (dko) mouse models by reintroduction of dystrophin has validated gene therapy as a potential therapeutic approach for Duchenne muscular dystrophy. However, the size of the dystrophin gene exceeds the capacity of adeno-associated viral (AAV) vectors. Dystrophin provides a mechanical link at the muscle membrane by direct binding of its amino-terminal and cysteine-rich domains to actin and a transmembrane protein complex, respectively. It has not been investigated whether restoration of these two tethering functions by two separate dystrophin molecules is sufficient to prevent dystrophic pathologies. We examine the effect of coexpression of the amino-terminal and cysteine-rich domains from separate dystrophin transgenes, Deltacys and Dp71, on the dystrophic phenotype. Expression of individual dystrophin domains from multiple vectors would effectively expand AAV capacity. Although both Deltacys and Dp71 colocalize at the membrane, there is no improvement of dystrophic pathology. The fiber-type and neuromuscular junction abnormalities of dko mice that are ameliorated by the Deltacys transgene are not further improved or disrupted by Dp71. Separate truncated dystrophins, which together restore all protein interactions and scaffolding for signaling molecules, are not sufficient to ameliorate the dystrophic phenotype and therefore dystrophin domains in trans cannot be used to increase the effective cloning capacity for AAV-mediated gene therapy. PMID- 16307001 TI - XIAP-mediated neuroprotection in retinal ischemia. AB - Retinal ischemia results in the loss of vision in a number of ocular diseases including acute glaucoma, diabetic retinopathy, hypertensive retinopathy and retinal vascular occlusion. Recent studies have shown that most of the neuronal death that leads to loss of vision results from apoptosis. XIAP-mediated gene therapy has been shown to protect a number of neuronal types from apoptosis but has never been assessed in retinal neurons following ischemic-induced cell death. We injected an adeno-associated viral vector expressing XIAP or GFP into rat eyes and 6 weeks later, rendered them ischemic by raising intraocular pressure. Functional analysis revealed that XIAP-treated eyes retained larger b-wave amplitudes than GFP-treated eyes up to 4 weeks post-ischemia. The number of cells in the inner nuclear layer (INL) and the thickness of the inner retina were significantly preserved in XIAP-treated eyes compared to GFP-treated eyes. Similarly, there was no significant reduction in optic nerve axon numbers in XIAP treated eyes. There were also significantly fewer TUNEL (TdT-dUTP terminal nick end labeling) positive cells in the INL of XIAP-treated retinas at 24 h post ischemia. Thus, XIAP-mediated gene therapy imparts both functional and structural protection to the retina after a transient ischemic episode. PMID- 16307002 TI - Activators of viral gene expression in polarized epithelial monolayers identified by rapid-throughput drug screening. AB - Epithelial polarity and tight junction formation limit the ability of adenovirus, retrovirus and adeno-associated virus (AAV) to deliver and express virally encoded genes. Using an extended half-life luciferase assay and high-throughput luminometry, we screened 23 000 compounds and natural product extracts as potentiators to overcome this barrier. Seven strong activators were discovered (up to several hundred fold above control) and two of these exhibited spectrum of activity in multiple cell types (HeLa (human cervical carcinoma), cystic fibrosis bronchial epithelial (human bronchial), HT29 (human colonic carcinoma), Calu3 (airway serous glandular)). Enhanced transduction by unrelated gene transfer vectors (adenovirus, lentivirus, AAV, liposomal) was also observed. These results establish a strategy for identifying compounds that improve viral gene transfer to resistant cell types, and provide new tools for examining epithelial defense against viral infection. The compounds should have broad usefulness in experimental therapies for cancer and genetic diseases. PMID- 16307003 TI - Hypoxia- and radiation-activated Cre/loxP 'molecular switch' vectors for gene therapy of cancer. AB - Although a significant negative prognostic factor, tumor hypoxia can be exploited for gene therapy. To maximize targeting within the tumor mass, we have developed synthetic gene promoters containing hypoxia-responsive elements (HREs) from the erythropoietin (Epo) gene as well as radiation-responsive CArG elements from the early growth response (Egr) 1 gene. Furthermore, to achieve high and sustained expression of the suicide gene herpes simplex virus thymidine kinase (HSVtk), our gene therapy vectors contain an expression amplification system, or 'molecular switch', based on Cre/loxP recombination. In human glioma and breast adenocarcinoma cells exposed to hypoxia and/or radiation, the HRE/CArG promoter rapidly activated Cre recombinase expression leading to selective and sustained HSVtk synthesis. Killing of transfected tumor cells was measured after incubation with the prodrug ganciclovir (GCV; converted by HSVtk into a cytotoxin). In vitro, higher and more selective GCV-mediated toxicity was achieved with the switch vectors, when compared with the same inducible promoters driving HSVtk expression directly. In tumor xenografts implanted in nude mice, the HRE/CArG switch induced significant growth delay and tumor eradication. In conclusion, hypoxia- and radiation-activated 'molecular switch' vectors represent a promising strategy for both targeted and effective gene therapy of solid tumors. PMID- 16307004 TI - Neonatal mortality and morbidity in vertex-vertex second twins according to mode of delivery and birth weight. AB - OBJECTIVE: To assess the risk of neonatal mortality and morbidity in vertex vertex second twins according to mode of delivery and birth weight. STUDY DESIGN: Data from a historical cohort study based on a twin registry in the US (1995 1997) were used. Multivariate logistic regression was used to control for maternal age, race, marital status, cigarette smoking during pregnancy, parity, medical complications, gestational age, and other confounders. RESULTS: A total of 86 041 vertex-vertex second twins were classified into two groups: second twins delivered by cesarean section after cesarean delivery of first twin (C-C) (43.0%), second twins whose co-twins delivered vaginally (V-X) (57.0%). In infants of birth weight>or=2500 g group, the risks of noncongenital anomaly related death (adjusted odds ratio (aOR): 4.64, 95% confidence interval (95% CI): 1.90, 13.92), low Apgar score (aOR: 2.39, 95% CI: 1.43, 4.14), and ventilation use (aOR: 1.31, 95% CI: 1.18, 1.47) were higher in the V-X group compared with the C-C group. No asphyxia-related neonatal deaths occurred in C-C group, whereas the incidence of this death was 0.04% in the V-X group. CONCLUSION: The risks of neonatal mortality and morbidity are increased in vertex-vertex second twins with birth weight>or=2500 g whose co-twins delivered vaginally compared with second twins delivered by cesarean section after cesarean delivery of first twin. PMID- 16307007 TI - Association between comorbidity and erectile dysfunction in patients with diabetes. AB - The aims of this study were to determine the prevalence of erectile dysfunction (ED) and its relationship with comorbidity in patients with diabetes. The study population comprised of 312 consecutive patients aged 20 years or over residing in the city of Hamadan in Iran in 2005. Depression was assessed by the modified version of the Beck Depression Inventory (BDI-II) and ED by the short form of the International Index of Erectile Function (IIEF-5) questionnaire. Potential confounding was controlled by stratification and by a logistic regression model. The prevalence of moderate or complete ED (IIEF score 450 x 10(9)/l, P=0.001), presence of one cardiovascular risk factor (arterial hypertension, smoking, hypercholesterolemia, or diabetes, P=0.003), cellular phase of myelofibrosis (P=0.005), and Hb >11 g/dl (P=0.02). Considering post-diagnosis events, the 5-year thrombosis-free survival probability was 90.4% in the series, 80.6% for patients with platelets >450 x 10(9)/l, 82.6% for patients with one cardiovascular risk factor, and 85.1% for those in cellular phase. These results indicate an increased thrombotic risk for IM patients with hyperproliferative features and/or coexistent cardiovascular risk factors. PMID- 16307012 TI - Peripheral T-cell non-Hodgkin's lymphoma NOS: naming of parts. PMID- 16307014 TI - Reply to 'hRap1B-retro: a novel human processed Rap1B gene blurs the picture?' by Zemojtel et al. PMID- 16307013 TI - Maintained immunogenicity of chronic myeloid leukemia-derived dendritic cells in the presence of imatinib mesylate: implication for vaccination regimens. PMID- 16307016 TI - HRap1B-retro: a novel human processed rap1B gene blurs the picture? PMID- 16307017 TI - The c-kit (CD117) sequence variation M541L, but not N564K, is frequent in the general population, and is not associated with CML in Caucasians. PMID- 16307018 TI - Allogeneic hematopoietic stem-cell transplantation in AML and MDS using myeloablative versus reduced-intensity conditioning: the role of dose intensity. AB - Allogeneic stem-cell transplantation (SCT) with both myeloablative and reduced intensity conditioning (RIC) is an effective therapy in AML/MDS. However, the relative merits of each may differ in different settings. To define the role of dose intensity, we analyzed SCT outcomes of 112 consecutive patients with AML/MDS. A total of 45 patients met eligibility criteria for standard myeloablative conditioning and were given intravenous-busulfan (12.8 mg/kg) and cyclophosphamide (ivBuCy). A total of 67 noneligible patients were given RIC with fludarabine and intravenous-busulfan (6.4 mg/kg, FB2, n=41) or a modified myeloablative regimen with fludarabine and myeloablative doses of intravenous busulfan (12.8 mg/kg, FB4, n=26). The overall survival (OS) at 2 years was 50, 49 and 47% after ivBuCy, FB4 and FB2, respectively (P=NS). Nonrelapse mortality was higher after ivBuCy, 22 vs 8% (P=0.05), but relapse rates were lower. Active disease at SCT was the most significant predictor of reduced survival in multivariable analysis (HR 4.5, P=0.0001). Myeloablative and RIC regimens had similar outcomes when leukemia was in remission at SCT; however, patients with active disease could only be salvaged by myeloablative conditioning. Among the latter, OS was 45% after ivBuCy but no FB2 recipient survived (P=0.02). Patients with active disease, ineligible for standard myeloablation, could tolerate modified myeloablation well; however, long-term outcome cannot be determined yet. PMID- 16307019 TI - Long-term clonal persistence and evolution of t(14;18)-bearing B cells in healthy individuals. PMID- 16307020 TI - Assessment of F-MuLV-induced tumorigenesis reveals new candidate tumor genes including Pecam1, St7, and Prim2. PMID- 16307021 TI - Nfl gene inactivation in acute myeloid leukemia cells confers cytarabine resistance through MAPK and mTOR pathways. PMID- 16307022 TI - MLL gene rearrangements have no direct impact on Ara-C sensitivity in infant acute lymphoblastic leukemia and childhood M4/M5 acute myeloid leukemia. PMID- 16307023 TI - The G(-248)A polymorphism in the promoter region of the Bax gene does not correlate with prognostic markers or overall survival in chronic lymphocytic leukemia. AB - The G(-248)A polymorphism in the promoter region of the Bax gene was recently associated with low Bax expression, more advanced stage, treatment resistance and short overall survival in B-cell chronic lymphocytic leukemia (CLL), the latter particularly in treated patients. To investigate this further, we analyzed 463 CLL patients regarding the presence or absence of the G(-248)A polymorphism and correlated with overall survival, treatment status and known prognostic factors, for example, Binet stage, VH mutation status and genomic aberrations. In this material, similar allele and genotype frequencies of the Bax polymorphism were demonstrated in CLL patients and controls (n=207), where 19 and 21% carried this polymorphism, respectively, and no skewed distribution of the polymorphism was evident between different Binet stages and VH mutated and unmutated CLLs. Furthermore, no difference in overall survival was shown between patients displaying the G(-248)A polymorphism or not (median survival 85 and 102 months, respectively, P=0.21), and the polymorphism did not influence outcome specifically in treated CLL. Neither did the polymorphism affect outcome in prognostic subsets defined by VH mutation status or genomic aberrations. In conclusion, the pathogenic role and clinical impact of the Bax polymorphism is limited in CLL. PMID- 16307024 TI - Expression of Tel-Aml1 transcript variants in pediatric acute lymphoblastic leukemia. PMID- 16307025 TI - Effects of 5-aza-2'-deoxycytidine on proliferation, differentiation and p15/INK4b regulation of human hematopoietic progenitor cells. AB - The demethylating agents 5-azacytidine and 5-aza-2'-deoxycytidine (DAC) have been shown to induce differentiation and inhibit growth of leukemic myeloid cells at low concentrations. However, the effect of DAC in changing the differentiation and proliferation behavior of normal human myeloid progenitors has rarely been investigated. Therefore, we established an in vitro model of normal hematopoietic differentiation, using CD34+ cells from mobilized peripheral blood, to study proliferation and colony formation, expression of several myeloid maturation markers and of the inhibitor of cyclin-dependent kinases p15/INK4b. Upon DAC treatment, cell growth was significantly decreased in a dose-dependent manner, without an increase in cytotoxicity. DAC treatment also resulted in a substantial increase of lysozyme-positive cells, which could be enhanced by G-CSF, a modest increase of myeloperoxidase+ and CD15+ cells, as well as an increase of colony forming cells (CFU-GM) compared to control cells. p15/INK4b protein expression was strongly upregulated upon myeloid maturation, and additional DAC treatment did not change p15 expression or the methylation status of the p15 promoter at the noncytotoxic concentrations used. Taken together, these data indicate a role of DAC in changing myeloid progenitor cell expansion and differentiation. This model appears suitable also for global analyses of multiple differentially methylated genes. PMID- 16307026 TI - Extramedullary relapse in acute promyelocytic leukemia treated with all-trans retinoic acid and chemotherapy. AB - We analyzed the incidence, presenting features, risk factors of extramedullary (EM) relapse occurring in acute promyelocytic leukemia (APL) treated with all trans retinoic acid (ATRA) and chemotherapy by using a competing-risk method. In total, 740/ 806 (92%) patients included in three multicenter trials (APL91, APL93 trials and PETHEMA 96) achieved CR, of whom 169 (23%) relapsed, including 10 EM relapses. Nine relapses involved the central nervous system (CNS) and one the skin, of which two were isolated EM relapse. In patients with EM disease, median WBC count was 26950/mm3 (7700-162000). The 3-year cumulative incidence of EM disease at first relapse was 5.0%. Univariate analysis identified age <45 years (P=0.05), bcr3 PML-RARalpha isoform (P= 0.0003) and high WBC counts (> or = 10,000/ mm3) (P<0.0001) as risk factors for EM relapse. In multivariate analysis, only high WBC count remained significant (P= 0.001). Patients with EM relapse had a poorer outcome since median survival from EM relapse was 6.7 months as compared to 26.3 months for isolated BM relapse (P=0.04). In conclusion, EM relapse in APL occurs more frequently in patients with increased WBC counts (> or = 10,000/mm3) and carries a poor prognosis. Whether CNS prophylaxis should be systematically performed in patients with WBC > or = 10,000/mm3 at diagnosis remains to be established. PMID- 16307027 TI - Determinants of hospital length of stay among patients with pneumonia admitted to a large Canadian hospital from 1991 to 2001. AB - BACKGROUND: Pneumonia is a common reason for hospital admission, and the cost of treatment is primarily determined by length of stay (LOS). OBJECTIVES: To explore the changes to and determinants of hospital LOS for patients admitted for the treatment of community-acquired pneumonia over a decade of acute hospital downsizing. METHODS: Data were extracted from the database of Vancouver General Hospital, Vancouver, British Columbia, on patients admitted with community acquired pneumonia (International Classification of Diseases, Ninth Revision, Clinical Modification codes 481.xx, 482.xx, 483.xx, 485.xx and 486.xx) from January 1, 1991 to March 31, 2001. The effects of sociodemographic factors, the specialty of the admitting physician (family practice versus specialist), admission from and/or discharge to a long-term care facility (nursing home) and year of admission, adjusted for comorbidity, illness severity measures and other potential confounders were examined. Longitudinal changes in these factors over the 10-year period were also investigated. RESULTS: The study population (n=2495) had a median age of 73 years, 53% were male and the median LOS was six days. Adjusted LOS was longer for women (10% increase, 95% CI 3 to 16), increasing age group (7% increase, 95% CI 4 to 10), admission under a family physician versus specialist (42% increase, 95% CI 32 to 52) and admission from home with subsequent discharge to a long-term care facility (75% increase, 95% CI 47 to 108). Adjusted hospital LOS decreased by an estimated 2% (95% CI 1 to 3) per annum. The mean age at admission and the proportion admitted from long-term care facilities both increased significantly over the decade (P<0.05). CONCLUSIONS: Results suggest that the management of hospitalized patients with pneumonia changed substantially between 1991 and 2001. The interface of long-term care facilities with acute care would be an important future area to explore potential efficiencies in caring for patients with pneumonia. PMID- 16307028 TI - Deep inspiration avoidance and airway response to methacholine: Influence of body mass index. AB - OBJECTIVE: To evaluate the effects of deep inspiration avoidance response to methacholine inhalation in 23 nonobese (body mass index between 18 kg/m2 and 30 kg/m2) and 27 obese (body mass index 30 kg/m2 or greater), nonatopic, nonasthmatic normal subjects. METHODS: Each subject had four methacholine challenges. In tests A and B, the first postmethacholine forced expiratory volume in 1 s (FEV1) was measured at 30 s and 3 min postinhalation, respectively; tests C and D were single-dose tests (using the final dose of test B), with the first postmethacholine FEV1 being obtained at 3 min, without (test C) or with (test D) 20 min of deep inspiration avoidance before inhalation. RESULTS: The mean provocative concentrations inducing a 20% fall in FEV1 on tests A and B were 80.6 mg/mL and 28.5 mg/mL (P<0.0001) in nonobese subjects, respectively, and 56.3 mg/mL and 21.5 mg/mL (P<0.0001) in obese subjects, respectively. No significant differences were observed in test A or B between control and obese subjects. Mean falls in FEV1 for tests C and D were 20.3% and 40.0% (P=0.0003) in nonobese subjects, respectively, and 18.5% and 23.6% (P>0.05) in obese subjects, respectively. CONCLUSIONS: As previously observed in patients with asthma, the present study found that nonasthmatic obese subjects had no increase in the fall in FEV1 after deep inspiration avoidance before methacholine, whereas nonobese subjects did, suggesting that obesity alters airway function. No significant changes were found between groups for symptom perception. PMID- 16307029 TI - Aspergillus-related lung disease. AB - Aspergillus is a ubiquitous dimorphic fungus that causes a variety of human diseases ranging in severity from trivial to life-threatening, depending on the host response. An intact host defence is important to prevent disease, but individuals with pre-existing structural lung disease, atopy, occupational exposure or impaired immunity are susceptible. Three distinctive patterns of aspergillus-related lung disease are recognized: saprophytic infestation of airways, cavities and necrotic tissue; allergic disease including extrinsic allergic alveolitis, asthma, allergic bronchopulmonary aspergillosis, bronchocentric granulomatosis and chronic eosinophilic pneumonia; and airway and tissue invasive disease -- pseudomembranous tracheobronchitis, acute bronchopneumonia, angioinvasive aspergillosis, chronic necrotizing aspergillosis and invasive pleural disease. A broad knowledge of these clinical presentations and a high index of suspicion are required to ensure timely diagnosis and treatment of the potentially lethal manifestations of aspergillus-related pulmonary disease. In the present report, the clinical, radiographic and pathological aspects of the various aspergillus-related lung diseases are briefly reviewed. PMID- 16307030 TI - Alveolar soft-part sarcoma presenting with eosinophilia and shunt. AB - Alveolar soft-part sarcoma (ASPS) is a rare soft tissue tumour found in young adults that usually arises in skeletal muscle and occurs most frequently in the lower limbs. Radiological and pathological findings of ASPS in a 34-year-old man who presented with increasing shortness of breath over a period of four to six weeks with peripheral blood eosinophilia, hypoxemia and a significant arteriovenous shunt are reported. The present article is the fourth report of eosinophilia in association with sarcoma, and the first involving ASPS. PMID- 16307031 TI - Respiratory effects of stimulation of the limbic cortex in rats and their modulation with serotonin. AB - Acute experiments on rats showed that the anterior cingulate gyrus contains 2 efferent regions of the functional respiratory system (inhibitory supragenual and excitatory infragenual areas). Stimulation of these cortical areas produced a respiratory effect, which depended on activity of serotoninergic mechanisms in the solitary tract nucleus. PMID- 16307032 TI - Effect of drugs with various mechanisms of action on propulsive activity of the small intestine. AB - We studied the effect of drugs with various mechanisms of action on propulsive activity of the small intestine in healthy rats. Blockade of the major inhibitory influences realized via nonadrenergic noncholinergic inhibitory effector neurons was not followed by stimulation of intestinal transit. Propulsive activity of the small intestine increased upon treatment with drugs, whose effects are realized via acetylcholine or acetylcholine and serotonin. PMID- 16307033 TI - Role of respiratory epithelium in the development of hyperreactivity of bronchial smooth muscles. AB - In guinea pigs sensitized with ovalbumin the respiratory epithelium lost its ability to modulate the responses of airway smooth muscles to histaminergic stimuli. Incubation of bronchial segments with IL-5 potentiated the contractile responses of bronchial smooth muscles to histamine in both intact and sensitized animals. Incubation of bronchial segments with IL-5 receptors moderated contractile activity of segments from sensitized pigs, but not in the segments from intact controls. PMID- 16307034 TI - Increase in slow afterhyperpolarization led to learning delay in DBA mice. AB - We showed that differences in learning capacity between DBA and C57Bl/6 mice correlates with differences in slow afterhyperpolarization amplitude in hippocampal CA1 pyramid neurons. In DBA mice learning capacity is lower, but the amplitude of slow afterhyperpolarizations higher than in C57Bl/6 mice. PMID- 16307035 TI - Adaptation to periodic hypoxia and hyperoxia improves resistance of membrane structures in heart, liver, and brain. AB - A novel principle of short-term periodic adaptive training by varying the oxygen level from hypo- to hyperoxia is substantiated both theoretically and experimentally. Short-term adaptation to hypoxia-normoxia produced a membrane protective effect in the heart and cerebral cortex, but increased the sensitivity to free radical oxidation and decreased the level of components of the antioxidant defense system in the liver. Hypo-hyperoxia adaptation produced a membrane-stabilizing effect in the heart, brain, and liver, which was more pronounced compared to the effect of hypoxia-normoxia training. In contrast to hypoxia-normoxia adaptation, in case of hypo-hyperoxia training the adaptive defense developed as early as 15 days after the start of training. PMID- 16307036 TI - Cooperative type of platelet hypersensitivity to ADP. AB - We found that gestosis is associated with platelet hypersensitivity to ADP. Cell P2X1 receptors exhibited a positive cooperative response to ADP (EC(50)=10.88+/ 3.70 nM, Hill constant n=2.59+/-0.50 rel. units). Cooperative binding of ADP to platelet P2X1 receptors was also observed during incubation of cells from pregnant women with isosorbide dinitrate. PMID- 16307037 TI - Proteinase-activated type 1 receptors are involved in the mechanism of protection of rat hippocampal neurons from glutamate toxicity. AB - Survival of cultured rat hippocampal neurons was estimated 4, 24, and 48 h after 15-min exposure to the toxic effect of glutamate under conditions of pre- or coincubation with 10 nM thrombin. Thrombin inhibited glutamate-induced apoptosis in neurons 24 and 48 h after treatment, but had no effect on necrosis. Selective peptide agonist of proteinase-activated type 1 receptors simulated, but receptor antagonist suppressed the neuroprotective effect of thrombin. Our results suggest that peptide antagonist of type 1 receptors play a role in the mechanisms of neuronal protection from glutamate toxicity. PMID- 16307038 TI - Effect of natural antioxidants on antioxidant activity and lipid peroxidation in eye tissue of rabbits with chemical burns. AB - Chemical eye burns were accompanied by free radical oxidation of lipids and dysfunction of the antioxidant protection system. The decrease in antioxidant activity of blood plasma reflected a generalized type of these processes. The concentration of thiobarbituric acid-reactive substances increased in blood plasma from rabbits with eye burns. The severity of disturbances was maximum at the stage of trophic disorders. Combination therapy with antioxidants and standard pharmaceuticals effectively inhibited lipid peroxidation in eye tissue of experimental animals. PMID- 16307039 TI - State of the nonspecific component of the immune system during combination therapy for experimental bile peritonitis. AB - Combined use of sodium hypochlorite and intravenous laser irradiation of the blood in the postoperation therapy of 24-h bile peritonitis improved recovery of engulfing and digestive activity of neutrophilic granulocytes in the early postoperation period. Enzyme activity of neutrophilic granulocytes in earlier postoperation periods returned to normal after treatment with sodium hypochlorite alone. PMID- 16307040 TI - Systemic administration of antibodies to glutamate increases seizure threshold for pentylenetetrazole. AB - We studied the effect of single intraperitoneal treatment with antibodies to glutamate on pentylenetetrazole-induced acute generalized epileptiform activity in C57Bl/6 mice. The antiepileptic effect was observed 1.5 and 24 h after administration of antibodies to glutamate in doses of 10, 25, and 50 mg/kg. This treatment increased the thresholds of clonic seizures and tonic phase of seizures with lethal outcome. PMID- 16307041 TI - Functional activity of the adrenal cortex in mice of opposite strains with candidiasis. AB - Experiments were performed on two opposite strains of mice differing in the basal blood glucocorticoid concentration in the blood. Infection of animals with C. albicans fungi led to the development of hypocorticism. C57Bl/6 and CBA mice with progressive candidiasis showed a persistent and phasic course of hypocorticism, respectively. PMID- 16307042 TI - Electron probe microanalysis of potassium and sodium in clonogenic culture of human neural stem cells. AB - The mean potassium and sodium concentrations and distribution of potassium in clonogenic culture of human neural stem cells (neurosphere) were estimated by means of electron probe microanalysis. High sodium concentration was typical of undifferentiated cells. Potassium was irregularly distributed in the test structure. Our results confirm published data on heterogeneous morphological structure of neurospheres. PMID- 16307043 TI - Molecular mechanisms of modified sensitivity of the adenylate cyclase signaling system to biogenic amines during streptozotocin-induced diabetes. AB - We demonstrated changes in the sensitivity of the adenylate cyclase signaling system to biogenic amines (adrenoceptor agonists and serotonin) underwent a change in skeletal muscles of rats with 30-day streptozotocin-induced diabetes. Isoproterenol had a less significant stimulatory effect on adenylate cyclase in diabetic rats. Hormonal signals via Gi proteins were suppressed in animals with diabetes, which determined a greater stimulatory effect of norepinephrine and serotonin on adenylate cyclase. Hormones less significantly increased guanosine triphosphate-binding activity of G proteins in diabetic rats, which reflects the impairment of their functional coupling with receptors. PMID- 16307044 TI - Role of Cu2+ in free radical oxidation of human serum albumin and L-tyrosine dipeptide with multicomponent metal-containing xenobiotic. AB - Cu(2+) entering the composition of multicomponent metal-containing xenobiotic in vitro initiated free radical oxidation of human serum albumin and L-tyrosine dipeptide. Oxidative modification was accompanied by the formation of derivatives of amino acids tyrosine and tryptophan and structural changes in human serum albumin. PMID- 16307045 TI - Succinate-based preparation alleviates manifestations of the climacteric syndrome in women. AB - Clinical placebo-controlled study of Enerlit-Clima (bioactive succinate-based food additive) a showed positive effect of the preparation on general clinical and psychoemotional manifestations of the climacteric syndrome. A trend to an increase in estradiol level in early pathological climacteric and normalization of the endometrial status were observed. PMID- 16307046 TI - Kinetics of interaction of local anesthetics with human serum albumin. AB - We studied kinetic parameters of interaction of local anesthetics (lidocaine, tetracaine, bupivacaine, and two novel agents with proved local anesthetic potency RU-353 and RU-1117) with human serum albumin. Complexation of local anesthetics with human serum albumin is a time-dependent and reversible process; equilibrium was attained within 1.5-4.5 h depending on chemical nature of local anesthetics. PMID- 16307047 TI - Effect of vepeside on the function of reproductive system in rats. AB - Experiments on adult rats showed that a single intravenous injection of antitumor drug vepeside in a MTD (maximum tolerable dose) reduced the reproductive status during periods corresponding to exposure of mature sex cells, spermatocytes, and spermatogonia in male rats and exposure of oocytes in ovulating, mature, and primordial follicles in female rats. Reduction of the male and female reproductive function manifested in increased antenatal mortality of the progeny. The toxic effects of the drug on mature male sex cells caused temporary partial infertility. PMID- 16307048 TI - Chronic cocaine injections attenuate behavioral response of kappa-opioid receptors to U-50,488H agonist. AB - Chronic injections of cocaine (20 mg/kg daily for 10 days) increase activity and decrease anxiety in male C57Bl/6j mice in comparison with animals chronically injected with normal saline. U-50,488H (kappa-opioid receptor agonist; 2.5 mg/kg) produced an anxiolytic effect in animals preinjected with normal saline and had no effect in animals chronically injected with cocaine. Presumably, chronic activation of dopaminergic systems caused by cocaine injections is paralleled by desensitization of kappa-opioid receptor system. PMID- 16307049 TI - Effect of prednisolone on secondary messenger metabolism in the lymphocytes from patients with acantholysis bullosa. AB - Prednisolone in therapeutic concentrations blocks Ca(2+) channels of lymphocyte plasma membranes and prevents arachidonic acid-induced Ca(2+) entry into the cells. Glucocorticoid virtually did not modulate arachidonic acid-stimulated release of Ca(2+) from intracellular stores. No appreciable effect of the hormone on mitogen-induced changes in the intracellular content of cAMP was detected. PMID- 16307050 TI - Potentiation of GABA-activated currents by imidazobenzoimidazole derivative RU 353 in isolated cerebellum Purkinje cells. AB - Voltage clamp and concentration jump experiments performed on Purkinje cells isolated from rat cerebellum showed that novel imidazobenzoimidazole derivative RU-353 increased the amplitude of GABA-activated chlorine current in a dose dependent manner (EC50=15 microM for the currents activated by 1 microM GABA). RU 353 shifted the GABA dose-response curve to the left, but produced no effect on the maximum response (EC50 in control and in the presence of 30 microM RU-353 were 6.9 and 2.0 microM, respectively). PMID- 16307051 TI - Methodological approaches to detection of Tamm-Horsfall protein. AB - A new test system for measuring Tamm-Horsfall protein was developed. The system consists of antigenic diagnostic agent and immune serum. The specificity and sensitivity of the reagents were determined. The system was tested on 52 urine samples from subjects without renal diseases. PMID- 16307052 TI - Ex vivo expansion of hemopoietic precursor cells on a sublayer treated with parathyroid hormone. AB - The kinetics of hemopoietic precursor cells was studied in cultures treated with parathyroid hormone in a concentration of 10(-7) M. Long-term culturing of bone marrow with parathyroid hormone did not change the number of mature cells, while the number of precursors forming colonies in semisolid media increased 7-fold and the number of cells forming cobblestone areas on day 28 increased 9-10-fold. After 24 h culturing of bone marrow cells on an irradiated sublayer pretreated with parathyroid hormone for 8 and 12 weeks, the number of early hemopoietic precursor cells forming cobblestone areas on day 28 of culturing increased 2-and 5.5-fold, respectively. The expression of Bmi-1 gene responsible for self maintenance of stem hemopoietic cells increased in cultures treated with parathyroid hormone. It seems that parathyroid hormone can be used for expansion of hemopoietic stem cells ex vivo, which is essential for their transplantation to patients. PMID- 16307053 TI - Effects of preparations modifying Th1/Th2 ratio on the incidence of clinical variants of chronic graft-versus-host reaction. AB - Induction of chronic graft-versus-host reaction in a semiallogenic DBA/2- (DBA/2xC57Bl/6) F1 system leads to the development of Th1- or Th2-dependent immunopathologies. Modification of the Th1/Th2 ratio during induction with preparations acting on the immune system cells via different mechanisms and shifting the Th1/Th2 balance towards Th2 (bisphenol A, pentoxifylline, muramyl dipeptide) increases the incidence of Th2-dependent autoimmune lupus-like glomerulonephritis. PMID- 16307054 TI - Tissue-specific changes in heme oxygenase activity and level of nonprotein thiols in C57Bl/6 mice after whole-body gamma-irradiation. AB - Heme oxygenase catalyzes heme degradation and is an important component of the antioxidant defense. Nonprotein thiols participate in redox regulation of heme oxygenase gene expression. Changes in heme oxygenase activity and levels of nonprotein thiols in the liver, lungs, and brain of C57Bl/6 mice were studied on days 1-7 after whole-body gamma-irradiation in a dose of 10 Gy. The maximum increase in heme oxygenase activity was observed in the liver (to 196% in females and to 250% in males) and was associated with an 8-fold increase in the level of heme oxygenase-1 (inducible form of the enzyme) mRNA. The increase in heme oxygenase activity was less pronounced in the lungs, while in the brain this parameter slightly decreased. Changes in the levels of nonprotein thiols were sex dependent: in the liver and lungs this parameter increased in females and decreased in males. PMID- 16307055 TI - Stimulation of cell component of the immune response activates exploratory behavior in senescence accelerated OXYS rats. AB - Suppression of the cell component of the immune system and open-field behavior developing in OXYS rats by the age of 3 months are regarded as manifestations of accelerated aging. Stimulation of cell-mediated immune response with BCG vaccine caused a dose-dependent increase of orientation and exploratory activity of OXYS rats in the open field test to virtually the same level as in Wistar rats. PMID- 16307056 TI - Effects of iEW synthetic peptide isomers on bone marrow colony-forming capacity in vivo. AB - Studies of cooperative effects of D- or L-isomers of iEW dipeptide and neuraminidase on the number of splenic CFU, comparison of the peptide effects on CD34+ cells and splenic CFU, and evaluation of the effects of iEW D- or L-isomers on parameters of T-cell activation showed that interactions of L-(iEW) and D (iEW) with CD34+ surface receptors are realized through the same mechanism, while their different biological effects on hemopoietic precursor cells in vivo can be explained by their different influence on T-component of the microenvironment. PMID- 16307057 TI - Morphofunctional peculiarities of the effects of electric coagulator on thyroid tissue. AB - Morphological changes in the thyroid tissue after exposure to coagulators were studied. Application of a monopolar coagulator resulted in the formation of narrow marginal coagulation necrosis on day 3, with a fibrous capsule of different thickness, and marginal sclerosis of the glandular parenchyma; inflammatory changes in the connective tissue in the resection edge persisted for 14 days. Bipolar coagulator produced the most pronounced damaging effect leaving septae up to 10 mm long penetrating into the depth of the gland and causing stubborn persistent deep and marginal sclerosis of the gland. PMID- 16307058 TI - Ultrastructural study of chitosan effects on Klebsiella and staphylococci. AB - Antibacterial effect of chitosan on the morphofunctional organization of clinical strains of Klebsiella pneumoniae and Staphylococcus aureus was studied by transmission electron microscopy. Chitosan promoted aggregation of bacterial cells and disorganization of bacterial cell wall and cytoplasmic membrane, which leads to the release of bacterial contents into the environment. These structural changes result in bacterial death. PMID- 16307059 TI - Effect of betulonic acid and its derivative [3-oxo-20(29)-lupene-28-oyl]-3 aminopropionic acid on liver structure in mice with RLS lymphoma. AB - Morphological study of the effects of semisynthetic derivatives of betulin (betulonic acid and [3-oxo-20(29)-lupene-28-oyl]-3-aminopropionic acid) on the liver of CBA/Lac mice with transplanted RLS lymphoma was studied in the control and after cytostatic polychemotherapy. The number of small focal necroses decreased, while the counts of hepatocytes in a state of slight hyaline droplet degeneration increased. Morphometry of the main elements of liver parenchyma showed that alanine amide derivative of betulonic acid decreases the severity of necrotic and degenerative changes in the liver parenchyma, induced by cytostatic polychemotherapy. Betulonic acid exhibited no appreciable hepatoprotective effect under these conditions. PMID- 16307060 TI - Mathematical model of binding of albumin-bilirubin complex to the surface of carbon pyropolymer. AB - We proposed a mathematical model and estimated the parameters of adsorption of albumin-bilirubin complex to the surface of carbon pyropolymer. Design data corresponded to the results of experimental studies. Our findings indicate that modeling of this process should take into account fractal properties of the surface of carbon pyropolymer. PMID- 16307061 TI - A simple method for isolation of cardiomyocytes from adult rat heart. AB - A simple, economic, and sparing method for isolation of cardiomyocytes from adult rat heart is proposed. Ultrastructure of cardiomyocytes from suspension of freshly isolated cells was studied by light and transmission electron microscopy. The isolated cardiomyocytes were viable, had characteristic shape and size, and retained their normal structure. PMID- 16307063 TI - Antimony: global environmental contaminant. PMID- 16307064 TI - A comparison of antimony and lead profiles over the past 2500 years in Flanders Moss ombrotrophic peat bog, Scotland. AB - Two cores collected in 2001 and 2004 from Flanders Moss ombrotrophic peat bog in central Scotland were dated (14C, 210Pb) and analysed (ICP-OES, ICP-MS) to derive and compare the historical atmospheric deposition records of Sb and Pb over the past 2500 years. After correction, via Sc, for contributions from soil dust, depositional fluxes of Sb and Pb peaked from ca. 1920-1960 A.D., with >95% of the anthropogenic inventories deposited post-1800 A.D. Over the past two centuries, trends in Sb and Pb deposition have been broadly similar, with fluctuations in the anthropogenic Sb/Pb ratio reflecting temporal variations in the relative input from emission sources such as the mining and smelting of Pb ores (in which Sb is commonly present, as at Leadhills/Wanlockhead in southern Scotland), combustion of coal (for which the Sb/Pb ratio is approximately an order of magnitude greater than in Pb ores) and exhaust emissions (Pb from leaded petrol) and abrasion products from the brake linings (Sb from heat-resistant Sb compounds) of automobiles. The influence of leaded petrol has been most noticeable in recent decades, firstly through the resultant minima in Sb/Pb and 206Pb/207Pb ratios (the latter arising from the use of less radiogenic Australian Pb in alkylPb additives) and then, during its phasing out and the adoption of unleaded petrol, complete by 2000 A.D., the subsequent increase in both Sb/Pb and 206Pb/207Pb ratios. The extent of the 20th century maximum anthropogenic enrichment of Sb and Pb, relative to the natural Sc-normalised levels of the Upper Continental Crust, was similar at approximately 50- to 100-fold. Prior to 1800 A.D., the influence of metallurgical activities on Sb and Pb concentrations in the peat cores during both the Mediaeval and Roman/pre-Roman periods was discernible, small Sb and Pb peaks during the latter appearing attributable, on the basis of Pb isotopic composition, to the mining/smelting of Pb ores indigenous to Britain. PMID- 16307065 TI - Lithogenic, oceanic and anthropogenic sources of atmospheric Sb to a maritime blanket bog, Myrarnar, Faroe Islands. AB - Antimony concentrations were measured in a core collected from Myrarnar, a blanket bog on the Faroe Islands which has been accumulating peat for more than six thousand years. The vertical distribution of Sb indicates that it has been supplied to the peat exclusively from the atmosphere. Despite the proximity to the ocean, the contribution of Sb to the peat from marine aerosols amounts to less than ca. 10% of the natural inputs. Although the peat core contains four notable layers of volcanic ash originating from Iceland, these have not contributed significantly to the Sb inventory. However, the distribution of Sb closely resembles that of Pb, with most of the Sb found in peats dating from the industrial period. Peat samples dating from the Roman Period are not only contaminated with Pb, but also with Sb. Lead is known to be immobile in peat bogs, and in Europe has been derived predominantly from industrial sources for thousands of years. The correlation between Sb and Pb in the peat core from the Faroe Islands supports the hypothesis that Sb is also effectively immobile in peat, and that ombrotrophic bogs are faithful archives of atmospheric Sb deposition. The data presented here also reinforces the view that natural Sb inputs during the past two centuries are dwarfed by industrial inputs, and that human activities have affected the atmospheric Sb cycle to a comparable extent to that of Pb. The natural rate of atmospheric Sb deposition recorded by the peat core (0.33 microg m(-2) year(-1)) is remarkably similar to the value obtained from a Swiss peat bog (Etang de la Gruere) in the samples dating from ca. 6000 to 9000 years ago (0.35 microg m(-2) year(-1)) which suggests that the background rates obtained from the peat cores have broader validity. Consistent with previous work, the data from the Faroe Islands suggests that the natural flux of Sb to the global atmosphere may have been overestimated by a factor of ten, and that the influence of human activities has been underestimated to the same extent. PMID- 16307066 TI - Concentrations, enrichment and predominant sources of Sb and other trace elements in size classified airborne particulate matter collected in Tokyo from 1995 to 2004. AB - APM was collected and trace elements existing in the particles were monitored since May 1995 in this study. APM sample was collected separately by size (d < 2 microm, 2-11 microm and >11 microm) on the roof of the university building (45 m above ground) in the campus of Faculty of Science and Engineering, Chuo University, Tokyo, Japan, using an Anderson low volume air sampler. The collected sample was digested by HNO3, H2O2 and HF using a microwave oven, and major elements (Na, Mg, Al, K, Ca and Fe) were measured by ICP-AES, and trace elements (Li, Be, Ti, V, Cr, Mn, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Sb, Ba and Pb) were measured by ICP-MS. It was observed that the APM concentration was higher between the winter and the spring, compared to during the summer. The enrichment factor was calculated for each element in each set of APM (d < 2 microm, 2-11 microm and >11 microm). Seasonal trends of enrichment factors were examined, and the elements were classified into 3 groups according to the common seasonal behavior. It is likely that the elements in the same group have common origins. Toxic pollutant elements (Sb, Se, Cd, Pb and As) were found in small particles with d of <2 microm in concentrated levels. Antimony (Sb) had the highest enrichment factor, and the results suggested that Sb level in APM was extremely high. The origins of Sb were sought, and wastes from plastic incineration and brake pad wears of automobiles were suspected. Each set of APM (d < 2 microm, 2-11 microm and >11 microm) was classified by the shape, and the shape-dependent constituents of a single APM particle were quantitatively measured by SEM-EDX. High concentration of Sb was found in APM <2 microm and square particles. Particles less than 2 microm and square shaped particles were major particles produced by actual car braking experiments. From these experimental results it was concluded that the source of Sb in squared APM <2 microm is considered to be from brake pad wear. PMID- 16307067 TI - Antimony: a traffic-related element in the atmosphere of Buenos Aires, Argentina. AB - Vehicular traffic is one of the main sources of antimony in highly populated urban areas like Buenos Aires where an overall traffic density of 1 500 000 vehicles per day (corresponding to 7500 vehicles km(-2)) is estimated. In this context, a study was undertaken to ascertain the levels of Sb and other traffic related elements (TRE) in the atmosphere of this city. To this end, sixty-seven samples of PM-10 particulate matter were collected during eight days in nine representative sampling sites located downtown Buenos Aires and spread over an area of about 30 km2. The collection of particulate matter was performed on ash free glass-fibre filters using high volume samplers with PM-10 sampling heads. A combination of aqua regia and perchloric acid was used for leaching metals from filters. The resulting solutions were evaporated and then diluted with 0.1 mol l( 1) HCl. Antimony was determined by inductively coupled plasma-quadrupole mass spectrometry (ICP-QMS) at ng g(-1) levels. Concentrations of Sb varied from 12.9 +/- 0.9 to 375 +/- 23 microg g(-1)(equivalent to 0.87 +/- 0.06 to 15.3 +/- 0.8 ng m(-3)). Statistical analysis was performed on the data set including the measured PM-10 mass and Sb concentrations for the monitored period. Correlations of Sb with other TRE namely, Cu and Mo were also assessed. The highest concentrations of Sb were detected at two sites (Hospital Aleman and Casa Rapallini) located in streets with traffic consisting mostly of passenger cars and showing a "stop-and go" pattern in peak hours. Antimony levels in the Buenos Aires PM-10 are by far below the level of 0.5 mg m(-3)(for an 8 hour workday, 40 hour work week) set by the US Occupational Safety and Health Administration (OSHA) for occupational exposure. However, monitoring of Sb and other TRE should be carried out in a systematic fashion to detect the possibility of increases in from the present levels. PMID- 16307068 TI - Increasing atmospheric antimony contamination in the northern hemisphere: snow and ice evidence from Devon Island, Arctic Canada. AB - Adopting recently developed clean laboratory techniques, antimony (Sb) and scandium (Sc) deposition were measured in a 63.72 m-long ice core (1842-1996) and a 5 m deep snow pit (1994-2004) collected on Devon Island, Canadian High Arctic. Antimony concentrations ranged from 0.07 to 108 pg g(-1) with a median of 0.98 pg g(-1)(N= 510). Scandium, used as a conservative reference element, revealed that dust inputs were effectively constant during the last 160 years. The atmospheric Sb signal preserved in the ice core reflects contamination from industrialisation, the economic boom which followed WWII, as well as the comparatively recent introduction of flue gas filter technologies and emission reduction efforts. Natural contributions to the total Sb inventory are negligible, meaning that anthropogenic emissions have dominated atmospheric Sb deposition throughout the entire period. The seasonal resolution of the snow pit showed that aerosols deposited during the Arctic winter, when air masses are derived mainly from Eurasia, show the greatest Sb concentrations. Deposition during summer, when air masses come mainly from North America, is still enriched in Sb, but less so. Snow and ice provide unambiguous evidence that enrichments of Sb in Arctic air have increased 50% during the past three decades, with two thirds being deposited during winter. Most Sb is produced in Asia, primarily from Sb sulfides such as stibnite (Sb2S3), but also as a by-product of lead and copper smelting. In addition there is a growing worldwide use of Sb in automobile brake pads, plastics and flame retardants. In contrast to Pb which has gone into decline during the same interval because of the gradual elimination of gasoline lead additives, the enrichments of Sb have been increasing and today clearly exceed those of Pb. Given that the toxicity of Sb is comparable to that of Pb, Sb has now replaced Pb in the rank of potentially toxic trace metals in the Arctic atmosphere. PMID- 16307069 TI - Adsorption of antimony(V) by floodplain soils, amorphous iron(III) hydroxide and humic acid. AB - Antimony (Sb) emissions to the environment are increasing, and there is a dearth of knowledge regarding Sb fate and behaviour in natural systems. In particular, there is a lack of understanding of sorption of the oxidised Sb(V) species onto soils and soil phases. In this study sorption of Sb(V) by two organic rich soils with high levels of oxalate extractable Fe was examined over the pH range of 2.5 7. Furthermore, the sorption behaviour of Sb(V) was examined in two phases mimicking those dominant in the experimental soils, namely a solid humic acid and an amorphous Fe(OH)3, across the same pH range. Sorption of Sb by the soils and the humic acid fitted a Freundlich type isotherm, with the equation parameters reflecting changes in bonding affinity corresponding to pH changes. The soils sorbed >75% of the added Sb in all trials, and 80-100% at pH values less than approximately 6.5. The Fe(OH)3 retained >95% of the added Sb in all experiments. The humic acid sorbed up to 60% of the added Sb at acidic pH values, but sorption decreased to zero at higher pH values. Further adsorption studies are recommended, such as examining the effects of ion competition and changes in ionic strength. PMID- 16307070 TI - Methylated arsenic, antimony and tin species in soils. AB - Methylated species of antimony, arsenic and tin were examined in urban soils of the Ruhr basin, near the cities of Duisburg and Essen, Germany. The main aim of this study was to investigate the occurrence of mono-, di- and trimethylated species of these elements in urban soils. The influence of historical and present land use upon the species content was examined. The distribution of inorganic As, Sb and Sn and their methylated species along the profile depth was investigated. As, Sb and Sn speciation was performed by pH-gradient hydride generation purge and trap gas chromatography, followed by inductively-coupled plasma mass spectrometry (HG-PT-GC/ICP-MS). Species' structures were confirmed by GC-EI/MS ICP-MS. Monomethylated Sb and As were the dominant species detected: the concentration of these metal(loid) species varied between <0.07-56 microg kg(-1) per dry mass. All dimethylated species and monomethyltin concentrations were between <0.01-7.6 microg kg(-1) per dry mass, and for the trimethylated species of all examined elements, concentrations between <0.001-0.63 microg kg(-1) per dry mass were detected. The highest organometal(loid) concentrations were observed in agricultural soils and garden soils; lower concentrations were found in the soils of abandoned industrial sites (wasteland, primary forest and grassland) and a flood plain soil of the Rhine. This result can be ascribed to both the cultivation and the increased biological activity of the agricultural soils, and the generally higher contamination, the disturbed structure and the artificial substrates (deposits from industrial sources) of the abandoned industrial soils. Due to periodical sedimentation, the flood plain profile was the only one where no depth dependence of organometal(loid) species concentration was detected. The other soil profiles showed a decrease of species content with increasing depth; this was particularly noticeable in soils with a clear change from a horizon with an organic character towards a mineral horizon, i.e. decreasing vitality from profile top to bottom. It is not as yet clear whether the organometal(loid) species are formed in the mineral horizons of the profiles or whether they are displaced from the organic, biologically-active horizons towards the mineral horizons. Field studies revealed that soil parameters like pH, water content or temperature did not correlate significantly with the degree of biomethylation observed. In contrast to the lower in vitro biomethylation efficiency of Sb vs. As in microbial incubations, we consistently detected higher proportions of transformed Sb compounds in situ in soil samples. These data may indicate a need to re-examine the currently accepted model of Sb biogeochemical cycling in the real environment. PMID- 16307071 TI - Investigation into antimony mobility in sewage sludge fermentation. AB - Antimony is distributed in the environment in inorganic and organic species with different solubility and mobility characters. Here we investigate the transformation of antimony in view of biomethylation during sewage sludge fermentation as a case study for an anaerobic environment. Our approach was to identify if antimony methylation follows the Challenger pathway by using isotopically enriched antimonite (123Sb(v)). The antimony source was subjected to methylation in sewage sludge, an anaerobic dominant methanogenic Archaea community. The antimony species were determined in the gas phase using cryotrapping (CT)-GC-ICP-MS, and in the medium (sewage slude) by hydride generation (HG) prior CT-GC-ICP-MS. The determined 123/121Sb isotope ratios in the volatile trimethylstibine and non-volatile methylantimony species indicated that the methylation follows the proposed methylation pathway. With this approach we were able to quantify 123Sb incorporation into monomethyl-, dimethyl- and trimethylantimony, respectively. The incorporation decreased with further methylation from 91% to 82% and 73%. Volatilisation as trimethystibine was generally lower than 0.1%, however, up to 0.8% of added antimony was found methylated to methylantimony species and mainly accumulated in the cell. Moreover, antimony biomethylation was enhanced by stimulation of the anaerobic communities of methanogenic Archaea and sulfate reducing bacteria (SRB), with the methanogens showing a higher activity. PMID- 16307072 TI - Antimony speciation in soil samples along two Austrian motorways by HPLC-ID-ICP MS. AB - Distribution of antimony and its inorganic species in soil samples along two traffic routes (A14, Rankweil and S36, Knittelfeld) in Austria was determined, since vehicle emissions are an important anthropogenic source of Sb in soil. The samples were taken along three parallel lines at about 0.2, 2 and 10 m distances from the edge of the road and in two depths range (0-5 and 5-10 cm from the soil surface). The optimized extraction was carried out using 100 mmol L(-1) citric acid at pH 2.08 applying an ultrasonic bath for 45 min at room temperature. Speciation analyses were done using on-line isotope dilution after a chromatographic separation of Sb species. Results of the two traffic routes confirmed significant accumulations of Sb at surface (0-5 cm depth) exceeding the natural background values by more than ten times at the S36 or four times at the A14. Concentrations of the extractable inorganic species decreased to natural background levels within a few meters from the edge of the traffic lane. The predominant Sb species was Sb(V). The Sb(III) concentrations at 5-10 cm depths range are nearly constant with distance from the edges of the two roads. Magnetic susceptibility data of all soil samples show the same distribution pattern as Sb and Sb(V) concentrations along the two traffic roads with an excellent correlation. This is an evidence for an anthropogenic source of Sb such as abrasions of motor vehicles surfaces or braking linings. The input of Sb and its inorganic species at one of the sampling sites (Knittelfeld) in samples taken in 2002 and in those taken recently (2005) was monitored. An increase in Sb (>or=30%), Sb(v)(>or=51%) and Sb(iii)(>or=10%) concentrations was only observed near the edge (> Zn >> Co > As >or= Mn. The fraction of Cu, Zn and As bound to humic acid (microg g(-1) of HA) increases in the first 10-12 cm of cores, then a decrease is observed. This observation could be related both to the presence of a mobile fraction of metals in sediments successively bound to humic acids and to different structural features showed by humic acids along the cores, due to transformation processes. For the other metals a variable pattern along the cores was found. PMID- 16307085 TI - Trace metals speciation in coastal particulate matter for marine environmental studies in Antarctica. AB - Solid speciation of some trace metals (Pb, Cd, Fe, Mn, Cu) having environmental relevance was studied in coastal particulate sampled during the Austral Spring 2000/2001. A nearshore station situated in the Gerlache Inlet of Terra Nova Bay (Ross Sea, Antarctica) was sampled from November to February. Samples were collected using the in situ filtration system FIS500, equipped with polycarbonate membrane filters having different pore sizes (10 microm, 2 microm and 0.4 microm) for the size fraction analysis of particles. The total concentration of metals was determined both in dissolved and particulate fractions, while speciation was determined on particulate by applying a sequential extraction procedure. Concerning the surface and sub-surface layers, it has been observed that concentration of elements is mainly affected by the dynamic of the pack ice melting and by phytoplankton activity. The solid speciation in November and December is similar for all the studied elements, while some differences can be noted in February, when the pack has completely melted and phytoplankton bloom occurs. With the exception of iron, during this sampling period the quantity of metal associated to the labile fraction increases. PMID- 16307086 TI - Homogeneity and stability study of the candidate reference material Adamussium colbecki for trace elements. AB - The preparation of a new candidate certified reference material (CRM) for trace elements based on the antarctic bivalve Adamussium colbecki(IRMM 813) was carried out by the Istituto Superiore di Sanita(ISS, Rome, Italy) in cooperation with the Institute for Reference Materials and Measurements, Joint Research Centre of the European Commission (EC-JRC-IRMM), in the frame of the Italian National Programme of Research in Antarctica (Programma Nazionale di Ricerca in Antartide, PNRA). Samples were collected at Terra Nova Bay (Ross Sea) during the 2000-2001 expedition in Antarctica. The preparation of a material suitable for certification was performed by the EC-JRC-IRMM. Measurements for homogeneity and short-term stability tests were carried out by ISS. The elements selected for the certification project were As, Cd, Cu, Cr, Fe, Mn, Ni and Zn. For the above mentioned tests, the determination of the elements of interest was performed on samples mineralised by microwave-assisted acid digestion. The analytical techniques employed to this end were inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma dynamic reaction cell quadrupole mass spectrometry (ICP-DRC-Q-MS). The candidate material was shown to be fit for purpose with regard to homogeneity and short-term stability, thus allowing the following phases of the certification project to be undertaken, in the first place the selection of expert laboratories for the accomplishment of the certification campaign. PMID- 16307087 TI - Formaldehyde determination in seawater. Preliminary application to coastal samples at Terra Nova Bay (Antarctica). AB - A sensitive spectrofluorimetric-FIA (flow injection analysis) method for formaldehyde (HCHO) determination was improved with the aim of analysing seawater samples. The fluorescence emission versus HCHO concentration shows a linear pattern from sub microg L(-1) to about 1000 microg L(-1). The reproducibility at 15 ppb level is about 2%. Interferences from other aldehydes were checked; only glyoxal shows a significative interference, but only when its concentration is about 6000 times higher than that of formaldehyde. Superficial (microlayer, just sub-pack or sea-ice free sea surface) and deep (along the water column, sub-pack or in sea-ice free areas) seawater samples were collected near the coast at Terra Nova Bay (Ross Sea, Antarctica) during the 1998/1999 and 2001/2002 Italian Antarctic Expedition. We report here the preliminary results of the spectrofluorimetric-FIA determination of the HCHO content. The mean seawater superficial formaldehyde concentration was 15 microg L(-1); the concentration along the water column ranged between 4.5 to over 40 microg L(-1)(20 microg L(-1) mean concentration), usually with a maximum value for the 30 m depth, corresponding to a fluorescence maximum. The sampling was repeated 7 times in the austral summer in order to evaluate seasonal changes in the formaldehyde concentration/seawater depth profiles. The results show changes in the formaldehyde concentration at different depths. PMID- 16307088 TI - Enrichment of organic pollutants in the sea surface microlayer (SML) at Terra Nova Bay, Antarctica: influence of SML on superficial snow composition. AB - Concentrations of dissolved and particle-associated n-alkanes, phthalates and polycyclic aromatic hydrocarbons (PAHs) were measured in sea surface microlayer (SML) and sub-surface water (SSL) samples collected in the coastal area of Terra Nova Bay, Antarctica, during the Austral spring 1998/1999. SML concentrations of the selected organic compounds were higher than SSL values and the enrichment factors were greater in the particulate phase than in the dissolved phase. During the same campaign, "fresh" snow samples, collected at different altitudes (from sea level up to 2670 m) near the coast on Mt Melbourne, immediately after a snowy event, were analysed in order to provide more information on air/sea exchange processes. The same classes of organic compounds found in sea water were also present in "fresh" snow samples. The surfactant fluorescent organic matter (SFOM), adsorbed on the microdrop aerosol surface, could be considered the main constituent of the enrichment and the carrier at higher altitudes of organic compounds. In fact, hydrocarbons (n-alkanes and PAHs), which are not surfactants like phthalates, could interact with SFOM and follow the same fate. PMID- 16307089 TI - Polychlorobiphenyls and polycyclic aromatic hydrocarbons in the sea-surface micro layer and the water column at Gerlache Inlet, Antarctica. AB - The enrichment of PCBs (polychlorobiphenyls) and PAHs (polycyclic aromatic hydrocarbons) in the sea-surface micro-layer and depth profile of these pollutants in the water column were investigated at Gerlache Inlet, Terra Nova Bay, Antarctica. Depth profile samplings were repeated three times during the Antarctic summer (from November to February). PCBs and PAHs showed a concentration range in the water column of 30-120 pg l(-1) and 150-400 pg l(-1), respectively, and these values were very much dependent on the suspended matter content. A nearly two-fold decrease in the pollutant concentration was also observed in the depth profile obtained in February, i.e. late summer, which might be correlated both with the high content of suspended matter and the reduction of the pollutant input. Moreover, isomer ratios of PAHs, such as LMW/HMW and PHE/ANT, highlight that the main PAH source might be petrogenic in nature, whereas the pyrolytic source seems to be less important. Sea surface micro-layer (SML) and sub-surface sea water (SSW) samples were simultaneously collected in the same site by a remote controlled rotating drum-based sampling system, a prototype named MUMS (Multi-User Micro-layer Sampler). Sea surface micro-layer samples showed a total content of PCBs and PAHs in the range 400-450 pg l(-1) and 2000-3000 pg l(-1), respectively, whereas the mean content of the sub-surface sea water samples was 48 pg l(-1) and 325 pg l(-1), respectively. The mean enrichment factors of PCBs and PAHs in sea-surface micro-layer were about 10 and 7, respectively. The surface excess concentrations of PCBs and PAHs were about 35 000 and 200 000, respectively. A fairly good correlation was observed between the concentration of pollutants and water solubility. Based on the assumption that POPs are confined in a very thin top layer of the SML about 0.01-0.001 microm thick, namely the sea-surface nano-layer, and also on an estimated thickness of the sampled sea-surface layer of about 100 microm, an enrichment factor of 10(5) 10(6) for the sea-surface nano-layer was calculated. Such a very high concentration increase was related to the two-fold increase of PAH concentration observed in the underlying 20 cm of the water column in late summer. PMID- 16307090 TI - Transport and modification of humic substances present in Antarctic snow and ancient ice. AB - We performed a study of fulvic acids extracted from fresh and aged snow, and from recent and ancient ice in Antarctica. The fresh snow samples were collected in coastal and inland sites to evaluate the influence of the distance from the sea on organic matter transport. Moreover, in a site (Melbourne Mountain) samples were collected at different heights to study the influence of altitude on transport. The obtained results showed that dissolved fulvic acid concentrations are influenced neither by distance nor by height while particulate fulvic acid concentrations are influenced by both parameters. Moreover, the results showed that fulvic acids transported for a long distance can undergo chemical modifications. Chemical modifications are better evidenced by the analysis of samples taken in trenches at different depth, which showed structural changes attributable to the loss of nitrogen-containing compounds and to an increase in aromatic character of the structures due to reduction and/or condensation processes. With ageing, the humification process proceeds with heavy carbon losses as demonstrated by results obtained from fulvic acids isolated from ice aged between twenty-five thousand and seventy thousand years. PMID- 16307091 TI - Glacial-interglacial changes in the occurrence of Pb, Cd, Cu and Zn in Vostok Antarctic ice from 240 000 to 410 000 years BP. AB - Lead (Pb), cadmium (Cd), copper (Cu) and zinc (Zn) have been measured by electrothermal atomic absorption spectrometry in various sections of the 3623 m deep ice core drilled at Vostok, in central East Antarctica. The sections were dated from 240 to 410 kyear BP (Marine Isotopic Stages (MIS) 7.5 to 11.3), which corresponds to the 3rd and 4th glacial-interglacial cycles before present. Concentrations are found to have varied greatly during this 170 kyear time period, with high concentration values during the coldest climatic stages such as MIS 8.4 and 10.2 and much lower concentration values during warmer periods, such as the interglacials MIS 7.5, 9.3 and 11.3. Rock and soil dust were the dominant sources for Pb, whatever the period, and for Zn and Cu and possibly Cd during cold climatic stages. The contribution from volcanic emissions was important for Cd during all periods and might have been significant for Cu and Zn during warm periods. PMID- 16307092 TI - Variations in the level of some trace elements in hair of participants in the Italian expeditions in Antarctica. AB - As a part the Italian National Programme of Research in Antarctica (PNRA) a monitoring study has been undertaken to quantify the concentrations of some selected trace elements in human hair of participants in the Antarctic expeditions. Such concentrations may vary as a consequence of the extreme environmental conditions and changes in lifestyle experienced by participants in the expeditions, as some evidence in previous investigations seems to suggest. The present study regards samples collected on the occasion of the 2002-2003 expedition to the Italian Base of Terra Nova Bay (now Mario Zucchelli Base), i.e., just before the expedition and about one month later. Seven essential elements were taken into account, namely, Ca, Cu, Cr, Fe, Mg, Mn and Mo. Determinations were performed by Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) and Dynamic Reaction Cell Inductively Coupled Plasma Mass Spectrometry (DRC-ICP-MS). Data obtained were statistically treated by using the non-parametric Friedman test. The concentrations of Ca, Cu and Mg were found to decrease (P < 0.05). The stress caused by the severe environmental conditions might well play a role in the observed decrease. PMID- 16307093 TI - Arsenic speciation in human hair: a new perspective for epidemiological assessment in chronic arsenicism. AB - The analysis for arsenic in hair is commonly used in epidemiological studies to assess exposure to this toxic element. However, poor correlation between total arsenic concentration in hair and water sources have been found in previous studies. Exclusive determination of endogenous arsenic in the hair, excluding external contamination has become an analytical challenge. Arsenic speciation in hair appears as a new possibility for analytical assessing in As-exposure studies. This study applied a relative simple method for arsenic speciation in human hair based on water extraction and HPLC-HG-ICP-MS. The concentration of arsenic species in human hair was assessed in chronically As(V)-exposed populations from two villages (Esquina and Illapata) of the Atacama Desert, Chile. The arsenic concentrations in drinking water are 0.075 and 1.25 mg L(-1), respectively, where As(V) represented between 92 and 99.5% of the total arsenic of the consumed waters. On average, the total arsenic concentrations in hair from individuals of Esquina and Illapata were 0.7 and 6.1 microg g(-1), respectively. Four arsenic species, As(III), DMA(V), MMA(V) and As(V), were detected and quantified in the hair extracts. Assuming the found species in extracts represent the species in hair, more than 98% of the total arsenic in hair corresponded to inorganic As. On average, As(III) concentrations in hair were 0.25 and 3.75 microg g(-1) in Esquina and Illapata, respectively; while, the As(V) average concentrations were 0.15 and 0.45 microg g(-1) in Esquina and Illapata, respectively. Methylated species represent less than 2% of the extracted As (DMA(V)+ MMA(V)) in both populations. As(III) in hair shows the best correlation with chronic exposure to As(V) in comparison to other species and total arsenic. In fact, concentrations of As(total), As(III) and As(V) in hair samples are correlated with the age of the exposed individuals from Illapata (R= 0.65, 0.69, 0.57, respectively) and with the time of residence in this village (R= 0.54, 0.71 and 0.58, respectively). PMID- 16307094 TI - Investigation on chemical species of arsenic, selenium and antimony in fly ash from coal fuel thermal power stations. AB - Investigations of the existing chemical forms and the concentrations of arsenic (As), selenium (Se) and antimony (Sb) in samples of fly ash obtained from six coal-fired power stations in various countries were carried out. The concentration of As was found to vary from 5.4 to 22.3 mg kg(-1), and the most common mode of occurrence of As in the fly ashes is in association with carbonates or Fe-Mn oxides. The concentrations of Se and Sb ranged from 2.3-5.2 and 1.0-3.9 mg kg(-1), respectively. The dominant chemical forms of Se and Sb in the fly ashes were as extractable species. Also, water-soluble As, Se and Sb in the fly ashes were extracted, and the chemical species of As, Se and Sb in the extract determined using HPLC-ICP-MS. This was done as the potential release of soluble As, Se and Sb through leaching of fly ash is of environmental concern. The most abundant form of As in the extract was the low toxicity As(V). The main species of Se was Se(IV), although it was found that the ratio of Se(VI) to Se(IV) in acidic fly ashes is higher than in alkaline fly ash samples. Antimony was mostly present as Sb(III). PMID- 16307095 TI - Impaction collection and slurry sampling for the determination of arsenic, cadmium, and lead in sidestream cigarette smoke by inductively coupled plasma mass spectrometry. AB - The slurry sampling technique has been applied for the determination of arsenic, cadmium and lead in sidestream cigarette smoke condensate (SS CSC) by inductively coupled plasma mass spectrometry (ICP-MS). The smoke collection system consists of a "fishtail" chimney in conjunction with an impaction trap and a mixed cellulose ester (MCE) backup filter for the SS CSC collection. The smoke condensates, collected at three different regions of the system (i.e. the inner wall of the chimney, the impaction trap and the MCE filter), were individually prepared and analyzed and showed that about 70 and 20% of Cd and Pb, respectively, were trapped by the impaction trap and the MCE filter with less than 5% trapped by the chimney. In contrast, about 60% of As was trapped by the impactor and a relatively high percentage of As, about 30%, was deposited onto the inner wall of the chimney. The uneven distribution of trace metals at each collection stage strongly suggested that the composition of smoke aerosol and the associated physical form of the analytes may be different. The yields of As, Cd and Pb for the sidestream cigarette smoke of the 1R4F reference cigarette were 29.5 +/- 2.4, 421 +/- 6.6 and 46.4 +/- 0.9 ng cigarette(-1)(n= 3) respectively, when the cigarette was smoked according to the smoking regime: 45 mL puff volume of 2-s puff duration at an interval of 30 s with 50% ventilation holes covered. The analytical results for the slurry sampling technique were also compared with other sample preparation techniques and show good agreement. PMID- 16307096 TI - Use of perlite in cadmium plant studies: an approach to polluted soil conditions. AB - Two different types of hydroponic cultures, "water culture" and "perlite system", were compared using white lupin plants (Lupinus albus L., cv. Marta) under different Cd treatments: 0, 0.2, 0.6, 2, 4, 6, 13, 20, 40 and 60 microM (water culture) and 0.2, 2, 20, 60 and 150 microM (moistened perlite). Fresh weight, shoot and root length, and total Cd concentration in the plants were measured. Moreover, a batch experiment was carried out to study the ability of perlite to adsorb and desorb Cd from nutrient solution. Lupin plants under Cd treatments in "water culture" showed a higher growth inhibition than those grown on perlite. A high positive correlation between Cd concentration in the plant and Cd supply was obtained regardless of the substrate used. Moreover, a high positive correlation between Cd doses with the "perlite system" and their equivalent Cd doses estimated for the "water culture" system was observed. Thus, the "water culture equivalent" Cd doses were 14 times lower than the Cd doses in the perlite system. On the other hand, desorbed Cd concentrations were calculated giving values 12 times lower than the tested Cd doses. PMID- 16307097 TI - Exposure and inhalation risk assessment in an aluminium cast-house. AB - To date the exposure, absorption and respiratory health effects of cast-house workers have not been described since most studies performed in the aluminium industry are focused on exposure and health effects of potroom personnel. In the present study, we assessed the external exposure and the absorbed dose of metals in personnel from the aluminium cast house. This was combined with an evaluation of respiratory complaints and the lung function of the personnel. 30 workers from an aluminium casting plant participated and 17 individuals of the packaging and distribution departments were selected as controls. The exposure was assessed by the quantification of total inhalable fume with metal fraction and by the determination of urinary aluminium, chromium, beryllium, manganese and lead concentration. Carbon monoxide (CO), carbon dioxide (CO2), aldehydes and polyaromatic hydrocarbons and man-made mineral fibres concentration were assessed as well. In order to evaluate their respiratory status each participant filled out a questionnaire and their lung function was tested by forced spirometry. Total inhalable fume exposure was maximum 4.37 mg m(-3). Exposure to the combustion gases, man-made mineral fibres and metal fume was well below the exposure limits. Beryllium could not be detected in the urine. The values of aluminium, manganese and lead in the urine were all under the respective reference value. One individual had a urinary chromium excretion above the ACGIH defined biological exposure index (BEI) of 30 microg g(-1) creatinine. There was no significant difference in any of the categories of the respiratory questionnaire and in the results of the spirometry between cast house personnel and referents (Chi-square, all p > 0.05). Exposure in cast houses seem to be acceptable under these conditions. However, peak exposure to fumes cannot be excluded and the potential risk of chromium and beryllium exposure due to the recycling of aluminium requires further attention. PMID- 16307098 TI - Spatial contaminant heterogeneity: quantification with scale of measurement at contrasting sites. AB - Material within the terrestrial environment is rarely homogeneously distributed, either spatially or temporally. One consequence of heterogeneity is that uncertainty is usually generated in measurements that are taken with the aim of characterising the environment. For example, a measurement of analyte concentration within soil taken from one sampling location on contaminated land can vary substantially when compared against another sample taken at effectively the same nominal location. The measurement uncertainty arising from the heterogeneity can substantially limit the reliability of the interpretations made upon environmental investigations. The sampling uncertainty usually outweighs the analytical uncertainty from the laboratory, often by a factor of 20 or more. One approach to reducing the uncertainty is to design a more suitable sampling strategy. This might be achieved by predicting the degree of heterogeneity prior to the investigation, but this is often difficult to achieve accurately. Another approach, which was investigated here, is to actually characterise the heterogeneity prior to the main investigation using rapid and inexpensive technology, such as in situ measurement techniques. In situ portable X-ray fluorescence (PXRF) and X-ray microprobe (XMP) techniques were employed to test the feasibility of this approach. Two contrasting contaminated land sites were chosen to characterise the two-dimensional spatial heterogeneity of heavy metal contamination in topsoil at a range of scales (50 m to 0.001 m). The spatial heterogeneity of contaminants, expressed as relative standard deviations, was found to differ between the two sites by a factor of two, largely due to the mode of deposition of pollution. The study also indicated that the heterogeneity did not change systematically with the scale of measurement between sampling locations at either site. PMID- 16307099 TI - Interlaboratory evaluation of endotoxin analyses in agricultural dusts- comparison of LAL assay and mass spectrometry. AB - Endotoxin exposure is associated with wheeze and asthma morbidity, while early life exposure may reduce risk of allergy and asthma. Unfortunately, it is difficult to compare endotoxin results from different laboratories and environments. We undertook this study to determine if lipopolysaccharide (LPS) extraction efficiency could account for differences among laboratories. We generated and collected aerosols from chicken and swine barns, and corn processing. We randomly allocated side-by-side filter samples to five laboratories for Limulus assay of endotoxin. Lyophilized aliquots of filter extracts were analyzed for 3-hydroxy fatty acids (3-OHFAs) as a marker of LPS using gas chromatography-mass spectrometry. There were significant differences in endotoxin assay and GC-MS (LPS) results between laboratories for all dust types (p < 0.01). Patterns of differences between labs varied by dust type. Relationships between assay and GC/MS results also depended on dust type. The percentages of individual 3-OHFA chain lengths varied across labs (p < 0.0001) suggesting that each lab recovered a different fraction of the LPS available. The presence of large amounts of particle associated LPS and absence of a freezing thawing cycle were associated with lower correlations between LPS and bioactivity, consistent with an absence of Limulus response to cell-bound endotoxin. These data suggest that extraction methods affect endotoxin measurements. The LAL methods may be most suitable when comparing exposures within similar environments; GC-MS offers additional information helpful in optimizing sample treatment and extraction. GC-MS may be of use when comparing across heterogeneous environments and should be considered for inclusion in future studies of human health outcomes. PMID- 16307100 TI - Organic contaminants in the Firth of Clyde following the cessation of sewage sludge dumping. AB - To assess contaminant concentrations in the Firth of Clyde in relation to the former sewage sludge dump site at Garroch Head, and investigate any temporal changes, fish have been sampled annually since 1992 and sediment since 1999. In addition, a further four locations in the Clyde (Holy Loch, Hunterston, Skelmorlie and Irvine Bay) have been sampled for fish and sediment since 1999. Chlorobiphenyls (CBs) were measured in fish samples and polycyclic aromatic hydrocarbons (PAHs) and CBs in sediment. Since sampling was initiated the concentration for the summation operatorICES7 CBs in fish liver has been consistently >500 microg kg(-1) lipid weight at Garroch Head and the other Clyde sites and lower at the reference sites (Pladda, Colonsay, Broad Bay). Although the lowest CB concentrations of the eleven year period in plaice liver from Garroch Head were found in 2002, CB concentrations were found not to have decreased significantly since sewage sludge dumping ceased in 1998. However, a change in CB profile was observed with fish liver collected between 1992 and 1998 from Garroch Head showing a lower proportion of the less chlorinated CBs compared to the 1999-2002 liver samples. Highest CB and PAH concentrations in sediment were found at Garroch Head and Holy Loch with concentrations at these sites being significantly higher than at all other sites. PMID- 16307101 TI - Bio-inspired energy security for planet Earth. PMID- 16307105 TI - Tunneling in PSII. AB - With available high resolution structures of PSII and a collection of reported redox midpoint potentials for most of the cofactors, it is possible to compare the expected electron tunneling rates with experimental rates to determine which electron transfer reactions are likely to reflect simply engineered electron tunneling, and which are more sophisticated and associated with large product rearrangements or the making and breaking of bonds. Reliable reorganization energies are largely lacking in this photosystem compared to PSI and purple bacteria and contribute about an order of magnitude uncertainty in tunneling rate estimates. Nevertheless it seems clear that as in purple bacterial reaction centers and PSI, with the notable exception of the oxygen evolving center, the majority of electron transfers within PSII are electron-tunneling limited at room temperature. Tunneling simulations also suggest that the short circuit between pheophytin and the adjacent chlorophyll cation may be fast enough to challenge triplet decay as the principle means of charge recombination from Q(A)(-) at room temperature. PMID- 16307106 TI - The mechanism of photosynthetic water splitting. AB - Oxygenic photosynthesis, which provides the biosphere with most of its chemical energy, uses water as its source of electrons. Water is photochemically oxidized by the protein complex photosystem II (PSII), which is found, along with other proteins of the photosynthetic light reactions, in the thylakoid membranes of cyanobacteria and of green plant chloroplasts. Water splitting is catalyzed by the oxygen-evolving complex (OEC) of PSII, producing dioxygen gas, protons and electrons. O(2) is released into the atmosphere, sustaining all aerobic life on earth; product protons are released into the thylakoid lumen, augmenting a proton concentration gradient across the membrane; and photo-energized electrons pass to the rest of the electron-transfer pathway. The OEC contains four manganese ions, one calcium ion and (almost certainly) a chloride ion, but its precise structure and catalytic mechanism remain unclear. In this paper, we develop a chemically complete structure of the OEC and its environment by using molecular mechanics calculations to extend and slightly adjust the recently-obtained X-ray crystallographic model with reference to this structure and to some important recent experimental results. PMID- 16307107 TI - Too much light? How beta-carotene protects the photosystem II reaction centre. AB - The photosystem II reaction centre of all oxygenic organisms is subject to photodamage by high light i.e. photoinhibition. In this review I discuss the reasons for the inevitable and unpreventable oxidative damage that occurs in photosystem II and the way in which beta-carotene bound to the reaction centre significantly mitigates this damage. Recent X-ray structures of the photosystem II core complex (reaction centre plus the inner antenna complexes) have revealed the binding sites of some of the carotenoids known to be bound to the complex. In the light of these X-ray structures and their known biophysical properties it is thus possible to identify the two beta-carotenes present in the photosystem II reaction centre. The two carotenes are both bound to the D2 protein and this positioning is discussed in relation to their ability to act as quenchers of singlet oxygen, generated via the triplet state of the primary electron donor. It is proposed that their location on the D2 polypeptide means there is more oxidative damage to the D1 protein and that this underlies the fact that this latter protein is continuously re-synthesised, at a far greater rate than any other protein involved in photosynthesis. The relevance of a cycle of electrons around photosystem II, via cytochrome b(559), in order to re-reduce the beta carotenes when they are oxidised and hence restore their ability to quench singlet oxygen, is also discussed. PMID- 16307108 TI - Photosynthesis: a blueprint for solar energy capture and biohydrogen production technologies. AB - Solar energy capture, conversion into chemical energy and biopolymers by photoautotrophic organisms, is the basis for almost all life on Earth. A broad range of organisms have developed complex molecular machinery for the efficient conversion of sunlight to chemical energy over the past 3 billion years, which to the present day has not been matched by any man-made technologies. Chlorophyll photochemistry within photosystem II (PSII) drives the water-splitting reaction efficiently at room temperature, in contrast with the thermal dissociation reaction that requires a temperature of ca. 1550 K. The successful elucidation of the high-resolution structure of PSII, and in particular the structure of its Mn(4)Ca cluster provides an invaluable blueprint for designing solar powered biotechnologies for the future. This knowledge, combined with new molecular genetic tools, fully sequenced genomes, and an ever increasing knowledge base of physiological processes of oxygenic phototrophs has inspired scientists from many countries to develop new biotechnological strategies to produce renewable CO(2) neutral energy from sunlight. This review focuses particularly on the potential of use of cyanobacteria and microalgae for biohydrogen production. Specifically this article reviews the predicted size of the global energy market and the constraints of global warming upon it, before detailing the complex set of biochemical pathways that underlie the photosynthetic process and how they could be modified for improved biohydrogen production. PMID- 16307109 TI - New vectors and marker excision systems mark progress in engineering the plastid genome of higher plants. AB - The transformation of the plastid genome, until recently restricted to tobacco, is now being extended to a rapidly growing list of crops. This perspective provides an overview of emerging trends of technology development in the field with a focus on vector design and marker excision systems. The new tools will facilitate engineering of the photosynthetic machinery and enable novel agricultural and industrial applications. PMID- 16307110 TI - Inactivation of photosynthetic electron flow during desiccation of desert biological sand crusts and Microcoleus sp.-enriched isolates. AB - Filamentous cyanobacteria, the main primary producers in biological sand crusts, survive harsh environmental conditions including diurnal desiccation/rehydration cycles. Here we describe the inactivation of photosystem II during dehydration of native crusts (NC) and Microcoleus sp. isolates grown on nitrocellulose filters (NCF). The morphology of NCF cells, visualized by scanning-transmission and atomic-force microscopy, disclosed long bacterial filaments encapsulated in extracellular polysaccharides (EPS) tubes consisting of parallel fibrils (100-400 nm wide and 50-100 nm high) oriented mostly perpendicular to the tube length. Presence of empty EPS tubes indicated a gliding capability of the cells. Desiccation of NC resulted in a rapid decline of F(o) and complete loss of F(v). These changes were accompanied by a decrease of 77 K PSII fluorescence emission relative to that of PSI, when excited at 430 nm, and a significant decrease of energy transfer from phycobilisomes to PSII. Lowering the turgor pressure through the addition of 1.5 M trehalose to natural crusts, reduced F(v)/F(m) by over 50% and was accompanied by a decrease of 77 K PSI fluorescence induced by chlorophyll excitation. Excitation of phycobilisomes resulted in a downshift of the PSI emission wavelength by 8 nm, indicative of reduced energy transfer from LHCI to the core PSI. Decline of F(v)/F(m) in trehalose-incubated NCF cells did not induce significant changes in 77 K fluorescence emission. These results suggest that alterations in energy transfer from antennae to reaction centers may be part of the survival strategy of Microcoleus. PMID- 16307111 TI - Quality control of Photosystem II: an FtsH protease plays an essential role in the turnover of the reaction center D1 protein in Synechocystis PCC 6803 under heat stress as well as light stress conditions. AB - The role of an AAA protease FtsH (slr0228) in the turnover of the D1 protein was studied under moderate heat stress conditions using wild-type cells of the cyanobacterium Synechocystis PCC 6803 and the mutant cells lacking a homologue of FtsH (slr0228). When the growth temperature of the wild-type was shifted from 30 degrees C to 40 degrees C, growth and oxygen-evolving activity were partially inhibited. Under the same heat stress, growth of the mutant was inhibited more significantly (63% inhibition after 5 days heat stress, compared with 26% inhibition with the wild-type cells) and the oxygen-evolving activity was also impaired in parallel. With heat stress at 42 degrees C, the level of the D1 protein of wild type cells was decreased, whereas that in mutant cells was not. The responses of cyanobacterial cells to heat stress observed here are quite similar to those to light stress that were reported previously. From these results, we suggest that the FtsH protease (slr0228) is responsible for both the heat-induced and light-induced degradation of the D1 protein. Notably, the amount of FtsH increased when the wild-type cells were exposed to heat stress or light stress, indicating that the up-regulation of the FtsH protease in the thylakoids is crucial for the cyanobacterial cells to cope with these abiotic stresses. PMID- 16307112 TI - Mutagenesis of CP43-arginine-357 to serine reveals new evidence for (bi)carbonate functioning in the water oxidizing complex of Photosystem II. AB - The chlorophyll-binding protein CP43 is an inner subunit of the Photosystem II (PSII) reaction center core complex of all oxygenic photoautotrophs. X-Ray structural evidence places the guanidinium cation of the conserved arginine 357 residue of CP43 within a few Angstroms to the Mn(4)Ca cluster of the water oxidizing complex (WOC) and has been implicated as a possible carbonate binding site. To test the hypothesis, the serine mutant, CP43-R357S, from Synechocystis PCC 6803 was investigated by PSII variable fluorescence (F(v)/F(m)) and simultaneous flash O(2) yield measurements in cells and thylakoid membranes. The R357S mutant assembles PSII-WOC centers, but is unable to grow photoautotrophically. Reconstitution of O(2) evolution by photoactivation and the occurrence of period-four oscillations of F(v)/F(m) establishes that the R357S mutant contains an assembled Mn(4)Ca cluster, but turnover is impaired as seen by an 11-fold larger Kok double miss parameter and faster decay of upper S states. Using pulsed light to avoid photoinactivation, wild-type cells and thylakoid membranes exhibit a 2-4-fold loss in O(2) evolution rate upon partial bicarbonate depletion under multiple turnover conditions, while the R357S mutant is unaffected by bicarbonate. Arginine R357 appears to function in binding a (bi)carbonate ion essential to normal catalytic turnover of the WOC. The quantum yield of electron donation from the WOC into PSII increases with decreasing turnover rate in R357S mutant cells and involves an aborted two-flash pathway that is distinct from the classical four-flash pattern. We speculate that an altered photochemical mechanism for O(2) production occurs via formation of hydrogen peroxide, by analogy to other treatments that retard the kinetics of proton release into the lumen. PMID- 16307113 TI - Conformational changes and their role in non-radiative energy dissipation in photosystem II reaction centres. AB - Accumulation of reduced pheophytin in photosystem II under illumination at low redox potential is known to be accompanied by a pronounced decrease of a chlorophyll fluorescence yield. Simultaneous measurement of this fluorescence quenching and absorbance changes in photosystem II reaction centres, in the presence of dithionite, showed each event to have a different temperature dependence. While fluorescence quenching was suppressed more than 20 times when measured at 77 K, pheophytin accumulation decreased only 5 times. At 77 K, the fluorescence was quenched considerably, but only in those reaction centres where reduced pheophytin had been accumulated at room temperature before sample freezing. This showed that the accumulation of reduced pheophytin above 240 K was accompanied by an additional, most probably conformational, change in the reaction centre that substantially enhanced non-radiative dissipation of excitation energy. PMID- 16307114 TI - Structure and evolution of the extrinsic proteins that stabilize the oxygen evolving engine. AB - PsbO, PsbP and PsbQ are the extrinsic proteins associated with the oxygen evolving (OE) engine of all known higher plants. However their presence is not constant throughout all known oxy-photosynthetic organisms. For this reason, comparative analyses of the sequence and the structure of these proteins in different species from prokaryotes to eukaryotes may allow unravelling of the evolutionary track that they have followed and infer new hints about their function in the OE complex. The results show that PsbP and PsbQ present different evolutionary profiles, and that PsbQ is more closely associated to PsbO and probably to the manganese stabilizing role assigned to this protein. PMID- 16307115 TI - Solving the structure of plant photosystem I--biochemistry is vital. AB - The recently determined structure of plant photosystem I (PSI) provides the first relatively high-resolution structural model of a supercomplex containing a reaction center and its peripheral antenna. Large amounts of highly purified PSI were required to get enough crystals amenable for structural determination by X ray crystallography. In addition, a deep biochemical understanding of the large supercomplex was vital for achieving the goal. The stability of PSI was analyzed by sucrose gradient centrifugation and gel electrophoresis. Small amounts of LHCI were detached from PSI following a 12 day incubation under crystallization conditions. The interaction between the reaction center and the peripheral antenna of PSI (LHCI) as well as the interactions among the LHCI monomers are flexible. Nevertheless, the pure and homogeneous preparation of PSI allows for relatively tight crystal packing, which holds promise for obtaining atomic resolution in the future. PMID- 16307116 TI - Excitation energy transfer from phycobiliprotein to chlorophyll d in intact cells of Acaryochloris marina studied by time- and wavelength-resolved fluorescence spectroscopy. AB - The fluorescence decay spectra and the excitation energy transfer from the phycobiliproteins (PBP) to the chlorophyll-antennae of intact cells of the chlorophyll (Chl) d-dominated cyanobacterium Acaryochloris marina were investigated at 298 and 77 K by time- and wavelength-correlated single photon counting fluorescence spectroscopy. At 298 K it was found that (i) the fluorescence dynamics in A. marina is characterized by two emission peaks located at about 650 and 725 nm, (ii) the intensity of the 650 nm fluorescence depends strongly on the excitation wavelength, being high upon excitation of phycobiliprotein (PBP) at 632 nm but virtually absent upon excitation of chlorophyll at 430 nm, (iii) the 650 nm fluorescence band decayed predominantly with a lifetime of 70 +/- 20 ps, (iv) the 725 nm fluorescence, which was observed independent of the excitation wavelength, can be described by a three-exponential decay kinetics with lifetimes depending on the open or the closed state (F(0) or F(m)) of the reaction centre of Photosystem II (PS II). Based on the results of this study, it is inferred that the excitation energy transfer from phycobiliproteins to Chl d of PS II in A. marina occurs with a time constant of about 70 ps, which is about three times faster than the energy transfer from the phycobilisomes to PS II in the Chl a-containing cyanobacterium Synechococcus 6301. A similar fast PBP to Chl d excitation energy transfer was also observed at 77 K. At 77 K a small long-lived fluorescence decay component with a lifetime of 14 ns was observed in the 640-700 nm spectral range. However, it has a rather featureless spectrum, not typical for Chl a, and was only observed upon excitation at 400 nm but not upon excitation at 632 and 654 nm. Thus, this long lived fluorescence component cannot be used as an indicator that the primary PS II donor of Acaryochloris marina contains Chl a. PMID- 16307117 TI - The assembly and organisation of photosynthetic membranes in Rhodobacter sphaeroides. AB - Recent AFM data demonstrate that mature photosynthetic membranes of R. sphaeroides are composed of rows of dimeric RC-LH1-PufX complexes with some LH2 complexes 'sandwiched' between these rows of core complexes, and others in discrete LH2-only domains which might form the light-responsive complement of the LH2 antenna. The present work applies membrane fractionation, radiolabelling and LDS-PAGE techniques to investigate the response of R. sphaeroides to lowered light intensity. The kinetics underlying this adaptation to low light conditions were revealed by radiolabelling with the bacteriochlorophyll (bchl) biosynthetic precursor, delta-aminolevulinate, which allowed us to measure only the bchls synthesised after the light intensity shift. We show that (1) the increase in LH2 antenna size is mainly restricted to the mature ICM membrane fraction, and the antenna composition of the precursor upper pigmented band (UPB) membrane remains constant, (2) the precursor UPB membrane is enriched in bchl synthase, the terminal enzyme of the bchl biosynthetic pathway, and (3) the LH2 and the complexes of intermediate migration in LDS-PAGE exhibit completely different labelling kinetics. Thus, new photosynthetic complexes, mainly LH2, are synthesised and assembled at the membrane initiation UPB sites, where the LH2 rings pack between the rows of dimeric cores fostering new LH2-LH1 interactions. Mature membranes also assemble new LH2 rings, but in this case the 'sandwich' regions between the rows of core dimers are already fully occupied and the bulk antenna pool is the favoured location for these new LH2 complexes. PMID- 16307118 TI - Isolation and characterization of a xanthophyll-rich fraction from the thylakoid membrane of Dunaliella salina(green algae). AB - Long-term acclimation to irradiance stress (HL) of the green alga Dunaliella salina Teod. (UTEX 1644) entails substantial accumulation of zeaxanthin along with a lowering in the relative amount of other pigments, including chlorophylls and several carotenoids. This phenomenon was investigated with wild type and the zea1 mutant of D. salina, grown under conditions of low irradiance (LL), or upon acclimation to irradiance stress (HL). In the wild type, the zeaxanthin to chlorophyll (Zea/Chl)(mol : mol) ratio was as low as 0.009 : 1 under LL and as high as 0.8 : 1 under HL conditions. In the zea1 mutant, which constitutively accumulates zeaxanthin and lacks antheraxanthin, violaxanthin and neoxanthin, the Zea/Chl ratio was 0.15 : 1 in LL and 0.57 : 1 in HL. The divergent Zea/Chl ratios were reflected in the coloration of the cells, which were green under LL and yellow under HL. In LL-grown cells, all carotenoids occurred in structural association with the Chl-protein complexes. This was clearly not the case in the HL-acclimated cells. A beta-carotene-rich fraction occurred as loosely bound to the thylakoid membrane and was readily isolated by flotation following mechanical disruption of D. salina. A zeaxanthin-rich fraction was specifically isolated, upon mild surfactant treatment and differential centrifugation, from the thylakoid membrane of either HL wild type or HL-zea1 mutant. Such differential extraction of beta-carotene and Zea, and their separation from the Chl-proteins, could not be obtained from the LL-grown wild type, although small amounts of Zea could still be differentially extracted from the LL-grown zea1 strain. It is concluded that, in LL-grown D. salina, xanthophylls (including most of Zea in the zea1 strain) are structurally associated with and stabilized by the Chl-proteins in the thylakoid membrane. Under HL-growth conditions, however, zeaxanthin appears to be embedded in the lipid bilayer, or in a domain of the chloroplast thylakoids that can easily be separated from the Chl-proteins upon mild surfactant treatment. In conclusion, this work provides biochemical evidence for the domain localization of accumulated zeaxanthin under irradiance-stress conditions in green algae, and establishes protocols for the differential extraction of this high-value pigment from the green alga D. salina. PMID- 16307119 TI - The mechanism for dioxygen formation in PSII studied by quantum chemical methods. AB - The availability of an X-ray structure for PSII including the water-oxidizing cluster, where the metal atoms and the amino acids are assigned, has opened up new possibilities to study the mechanism for dioxygen formation. In the present paper the main results of an ongoing hybrid DFT study are presented. The model used follows the structure suggested by the X-ray analysis as closely as possible. After nearly one thousand optimizations of different structures, each one with about 70 atoms, the main features of a water oxidizing mechanism start to emerge. The key intermediate is an oxyl radical state in S(3), stabilized by a weak trans effect to a bridging oxo in the cube. To reach this radical state a structural rearrangement appears necessary, in which one additional bridging oxo is formed between the dangling manganese and a manganese in the cube. The calculated energetics is reasonable but still not fully consistent with a correct mechanism. It is suggested that some part of the structure is not correct, probably the presence of the bicarbonate. PMID- 16307120 TI - A role of the C-terminal extension of the photosystem II D1 protein in sensitivity of the cyanobacterium Synechocystis PCC 6803 to photoinhibition. AB - The D1 protein, a key protein subunit of Photosystem II complex (PSII), is synthesised as a precursor (pD1) with a carboxyl-terminal extension. In the cyanobacterium Synechocystis sp. PCC 6803, this extension consists of 16 amino acid residues and it is cleaved by a specific protease in two putative steps with the final cleavage after the residue Ala344. In order to define the importance of the extension for the functioning of PSII, we constructed and characterized several site-directed mutants of Synechocystis that differ in the length and amino acid sequence of this extension. The mutant lacking the entire C-terminal extension exhibited slightly increased sensitivity to photoinhibition. Analysis of the PSII assembly in the mutant by the blue-native electrophoresis in combination with radioactive labelling revealed an increased level of the unassembled D1 protein in this strain. Replacement of the amino acid residue Asn359 by His or Asp also led to the higher vulnerability to photoinhibition of both mutants. In the Asn359His mutant, this vulnerability was accompanied by an increased level of the PSII core lacking CP43 indicating limitation of the repair cycle in the CP43 reassembly step. PMID- 16307121 TI - The role of D1-Ala344 in charge stabilization and recombination in Photosystem II. AB - The Ala344 residue of the D1 protein has been identified as a crucial residue of the catalytic cluster of the water-oxidizing complex, however, its function has not been fully clarified. Here we have used thermoluminescence and flash-induced chlorophyll fluorescence measurements to characterize the effect of the D1 Ala344stop mutation on the electron transport of Photosystem II in intact cells of the cyanobacterium Synechocystis 6803. Although the mutant cannot grow photoautotrophically it shows flash-induced thermoluminescence and chlorophyll fluorescence signals reflecting the stabilization of negative and positive charges on the Q(A) and Q(B) quinone electron acceptors, and stable Photosystem II donors, respectively. Decay of flash induced chlorophyll fluorescence yield is multiphasic in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), with 6 ms, 350 ms, and 26 s time constants. When cells are illuminated with repetitive flashes, fired at 1 ms intervals, the 6 ms phase is gradually decreased with the concomitant increase of the 350 ms phase. After 45 min dark adaptation of mutant cells the 6 ms and 350 ms phases were significantly decreased and a very slow decaying component was formed. Flash induced oscillation of the thermoluminescence B band, which reflects the redox cycling of the water-oxidizing complex in the wild-type cells, was completely abolished in the D1-Ala344stop mutant. The results demonstrate that low efficiency photooxidation of Mn occurs in about 60% of the PSII centers. The photooxidizable Mn is unstable in the dark, and formation of higher S states is inhibited. In addition, the Q(A) to Q(B) electron transfer step is slowed down as an indirect consequence of the donor side modification. Our data indicate that the stabilization of a Mn ion by the alpha-carboxylate chain of the D1-Ala344 residue might represent one of the final steps in the assembly of functional catalytic sites for water oxidation. PMID- 16307122 TI - Kinetic characterisation of the light-driven protochlorophyllide oxidoreductase (POR) from Thermosynechococcus elongatus. AB - The light-driven enzyme NADPH:protochlorophyllide oxidoreductase (POR) catalyses the reduction of the C17-C18 double bond of protochlorophyllide (Pchlide) to chlorophyllide (Chlide), which is a key regulatory step in the chlorophyll biosynthesis pathway. POR from the thermophilic cyanobacterium Thermosynechococcus elongatus is an attractive system for following the reaction and in the present work we have carried out a detailed steady state kinetic characterisation of this enzyme. The thermophilic POR was shown to have maximal activity at approximately 50 degrees C, which is similar to the growth temperature of the organism. The V(max) was calculated to be 0.53 microM min(-1) and the K(m) values for NADPH and Pchlide were 0.013 microM and 1.8 microM, respectively. The binding properties for both substrates as well as the NADP(+) product have been analysed by using fluorescence emission measurements, which have allowed the dissociation constants for binding to be calculated. These results represent the first steady state kinetic characterisation of a thermophilic version of POR. PMID- 16307123 TI - The nature of the photosystem II reaction centre in the chlorophyll d-containing prokaryote, Acaryochloris marina. AB - Pigment-protein complexes enriched in photosystem II (PS II) have been isolated from the chlorophyll (Chl) d containing cyanobacterium, Acaryochloris marina. A small PS II-enriched particle, we call 'crude reaction centre', contained 20 Chl d, 0.5 Chl a and 1 redox active cytochrome b-559 per 2 pheophytin a, plus the D1 and D2 proteins. A larger PS II-enriched particle, we call 'core', additionally bound the antenna complexes, CP47 and CP43, and had a higher chlorophyll per pheophytin ratio. Pheophytin a could be photoreduced in the presence of a strong reductant, indicating that it is the primary electron acceptor in photosystem II of A. marina. A substoichiometric amount of Chl a (less than one chlorophyll a per 2 pheophytin a) strongly suggests that Chl a does not have an essential role in the photochemistry of PS II in this organism. We conclude that PS II, in A. marina, utilizes Chl d and not Chl a as primary electron donor and that the primary electron acceptor is one of two molecules of pheophytin a. PMID- 16307124 TI - Degradation of the main Photosystem II light-harvesting complex. AB - Many factors trigger the degradation of proteins, including changes in environmental conditions, genetic mutations, and limitations in the availability of cofactors. Despite the importance for viability, still very little is known about protein degradation and its regulation. The degradation of the most abundant membrane protein on Earth, the light-harvesting complex of Photosystem II (LHC II), is highly regulated under different environmental conditions, e.g. light stress, to prevent photochemical damage of the reaction center. However, despite major effort to identify the protease/proteases involved in the degradation of the apoproteins of LHC II the molecular details of this important process remain obscure. LHC II belongs to the family of chlorophyll a/b binding proteins (CAB proteins) and is located in the thylakoid membrane of the plant chloroplast. The results of biochemical experiments to isolate and characterize the protease degrading LHC II are summarized here and compared to our own recent finding indicating that a metalloprotease of the FtsH family is involved in this process. PMID- 16307125 TI - Functional flexibility and acclimation of the thylakoid membrane. AB - Light is an elusive substrate for the function of photosynthetic light reactions of photosynthesis in the thylakoid membrane. Therefore structural and functional dynamics, which occur in the timescale from seconds to several days, are required both at low and high light conditions. The best characterized short-time regulation mechanism at low light is a rapid state transition, resulting in higher absorption cross section of PSI at the expense of PSII. If the low light conditions continue, activation of the lhcb-genes and synthesis of the light harvesting proteins will occur to optimize the functions of PSII and PSI. At high light, the transition to state 2 is completely inhibited, but the feedback de excitation of absorbed energy as heat, known as the energy-dependent quenching (q(E)), is rapidly set up. It requires, at least, the DeltapH-dependent activation of violaxanthin de-epoxidase and involvement of the PsbS protein. Another crucial mechanism for protection against the high light stress is the PSII repair cycle. Furthermore, the water-water cycle, cyclic electron transfer around PSI and chlororespiration are important means induced under high irradiation, functioning mainly to avoid an excess production of reactive oxygen species. PMID- 16307126 TI - Granal stacking of thylakoid membranes in higher plant chloroplasts: the physicochemical forces at work and the functional consequences that ensue. AB - The formation of grana in chloroplasts of higher plants is examined in terms of the subtle interplay of physicochemical forces of attraction and repulsion. The attractive forces between two adjacent membranes comprise (1) van der Waals attraction that depends on the abundance and type of atoms in each membrane, on the distance between the membranes and on the dielectric constant, (2) depletion attraction that generates local order by granal stacking at the expense of greater disorder (i.e. entropy) in the stroma, and (3) an electrostatic attraction of opposite charges located on adjacent membranes. The repulsive forces comprise (1) electrostatic repulsion due to the net negative charge on the outer surface of thylakoid membranes, (2) hydration repulsion that operates at small separations between thylakoid membranes due to layers of bound water molecules, and (3) steric hindrance due to bulky protrusions of Photosystem I (PSI) and ATP synthase into the stroma. In addition, specific interactions may occur, but they await experimental demonstration. Although grana are not essential for photosynthesis, they are ubiquitous in higher plants. Grana may have been selected during evolution for the functional advantages that they confer on higher plants. The functional consequences of grana stacking include (1) enhancement of light capture through a vastly increased area-to-volume ratio and connectivity of several PSIIs with large functional antenna size, (2) the ability to control the lateral separation of PSI from PSII and, therefore, the balanced distribution of excitation energy between two photosystems working in series, (3) the reversible fine-tuning of energy distribution between the photosystems by State 1-State 2 transitions, (4) the ability to regulate light harvesting via controlled thermal dissipation of excess excitation energy, detected as non-photochemical quenching, (5) dynamic flexibility in the light reactions mediated by a granal structure in response to regulation by a trans thylakoid pH gradient, (6) delaying the premature degradation of D1 and D2 reaction-centre protein(s) in PSII by harbouring photoinactived PSIIs in appressed granal domains, (7) enhancement of the rate of non-cyclic synthesis of adenosine triphosphate (ATP) as well as the regulation of non-cyclic vs. cyclic ATP synthesis, and (8) the potential increase of photosynthetic capacity for a given composition of chloroplast constituents in full sunlight, concomitantly with enhancement of photochemical efficiency in canopy shade. Hence chloroplast ultrastructure and function are intimately intertwined. PMID- 16307127 TI - Photosystem I: a search for green plant trimers. AB - Recent blue-native gel electrophoresis studies gave evidence for the existence of dimeric and trimeric PSI complexes in green plants. We used single particle electron microscopy to investigate all the larger particles from the thylakoid membrane of pea (Pisum sativum var. Charmette). Peak fractions with monomeric, dimeric and trimeric Photosystem I were obtained after solubilization with digitonin and size-exclusion chromatography. The analysis showed that only a few percent of dimers and trimers were present. In the best resolved trimers some of the monomers were oriented upside down. Many classes were fuzzy, indicating a non specific or flexible orientation. From these results we conclude that the green plant PSI is monomeric within the green plant membrane. PMID- 16307128 TI - Synthesis of classes of ternary metal oxide nanostructures. AB - Nanoscale structures, such as nanoparticles, nanorods, nanowires, nanocubes, and nanotubes, have attracted extensive synthetic attention as a result of their novel size-dependent properties. Ideally, the net result of nanoscale synthesis is the production of structures that achieve monodispersity, stability, and crystallinity with a predictable morphology. Many of the synthetic methods used to attain these goals have been based on principles derived from semiconductor technology, solid state chemistry, and molecular inorganic cluster chemistry. We describe a number of advances that have been made in the reproducible synthesis of various ternary oxide nanomaterials, including alkaline earth metal titanates, alkali metal titanates, bismuth ferrites, ABO(4)-type oxides, as well as miscellaneous classes of ternary metal oxides. PMID- 16307129 TI - Supramolecular click chemistry for the self-assembly of a stable Zn(II)-porphyrin C60 conjugate. AB - Owing to the complementarity between a bis-Zn(II)-porphyrin receptor and a fullerene ligand bearing two pyridine substituents, the substrate can be clicked onto the ditopic receptor, thus leading to a stable non-covalent macrocyclic 1 ratio 1 complex. PMID- 16307130 TI - Tumor angiogenic vasculature targeting with PAMAM dendrimer-RGD conjugates. AB - PAMAM dendrimer-RGD-4C peptide conjugate was synthesized and in vitro targeting efficacy to integrin receptor expressing cells was studied by flow cytometry and confocal microscopy. PMID- 16307131 TI - Allosteric function facilitates template assisted olefin metathesis. AB - Template assisted olefin metathesis of an allosteric host to give the corresponding bicyclic compound was achieved and can allosterically bind the template guest diamines, and with different affinity and cooperativity. PMID- 16307132 TI - The effect of selenium incorporation on the bandwidth and conductivity of neutral radical conductors. AB - The first example of an undimerized pi-stacked bis-1,2,3-thiaselenazolyl radical displays improved bandwidth and conductivity relative to an isostructural bis 1,2,3-dithiazolyl. PMID- 16307133 TI - Reversing the code of a template-directed solid-state synthesis: a bipyridine template that directs a single-crystal-to-single-crystal [2 + 2] photodimerisation of a dicarboxylic acid. AB - A linear template in the form of a bipyridine has been developed and is shown to direct a single-crystal-to-single-crystal [2 + 2] photodimerisation of a dicarboxylic acid. PMID- 16307134 TI - Hydrogen bond directed synthesis of pyridazine and naphthyridine containing macrocycles. AB - This work describes a high-yielding, one-step synthesis of pyrizadine and naphthyridine containing macrocycles directed by intramolecular H-bonding. PMID- 16307135 TI - Temperature-induced abrupt volume inflation in the mixed-valence ternary Zintl phase Yb8Ge3Sb5. AB - The Zintl phase, Yb8Ge3Sb5 exhibits a complex lattice response and an abrupt negative thermal expansion below 15 K - subtle structural changes before and after the transition are consistent with temperature-induced electron transfer from (to) Yb 4f bands to (from) Sb 5p and Ge 4p bands. PMID- 16307136 TI - Measuring rate constants for active species in the polymerization of ethylene by MAO-activated metallocene catalysts by electrospray ionization mass spectrometry. AB - Electrospray ionization mass spectrometry (ESI-MS) of MAO-activated metallocene catalyzed polymerizations quenched with carbodiimides provides a method for the determination of rate constants in Ziegler-Natta polymerizations. PMID- 16307137 TI - Chelated heteroatoms in polyoxometalates and the topological equivalence of [CoIII(en)] to type II cis-dioxometal centers. Synthesis and structure of [[Co(en)(mu-OH)2Co(en)}[PW10O37Co(en)]2]8- and [K subset[Co(en)WO4][WO(H2O)](PW9O34)2]12-. AB - The first examples of polyoxometalate structures that incorporate embedded chelated heteroatoms point to new possibilities for stereochemical control of applications. PMID- 16307138 TI - Dynamic sol-gel interconversion by reversible cation binding and release in G quartet-based supramolecular polymers. AB - The bis-guanine monomer G-G forms highly cross-linked, K+ stabilized, polymeric hydrogels that can be reversibly interconverted between gel and sol state via sequential binding and release of K+ by a cryptand undergoing protonation/deprotonation. PMID- 16307139 TI - Poly(9,9-dialkyl-3,6-dibenzosilole)--a high energy gap host for phosphorescent light emitting devices. AB - The preparation of the 3,6-disubstituted dibenzosilole monomers , and by two different routes is described; Suzuki copolymerisation afforded poly(9,9-dioctyl 3,6-dibenzosilole) which has a sufficiently high triplet energy (2.55 eV) to function as a host for green electrophosphorescent emitters. PMID- 16307140 TI - High-yield one-step synthesis in water of [Pt3n(CO)6n]2- (n > 6) and [Pt38(CO)44]2-. AB - Carbonylation of Na2PtCl6.6H2O, as well as K2PtCl6, in water under a CO pressure of 900 mm Hg selectively and quantitatively affords [Pt3n(CO)6n]2- (n > 6) salts; conversely, their corresponding carbonylation at reduced CO pressure of 760-800 mm Hg leads to a convenient one-step synthesis of [Pt38(CO)44]2-. PMID- 16307141 TI - Unprecedented reactivity in the Morita-Baylis-Hillman reaction; intramolecular alpha-alkylation of enones using saturated alkyl halides. AB - sp3 Hybridized electrophiles, never before used in the organomediated Morita Baylis-Hillman reaction, now facilitate the formation of five- and six-membered enone cycloalkylation products. PMID- 16307142 TI - Multivalent, bifunctional dendrimers prepared by click chemistry. AB - Unsymmetrical dendrimers, containing both mannose binding units and coumarin fluorescent units, have been prepared using click chemistry and shown to be highly efficient, dual-purpose recognition/detection agents for the inhibition of hemagglutination. PMID- 16307143 TI - Expanding functionality of RNA: synthesis and properties of RNA containing imidazole modified tandem G-U wobble base pairs. AB - Imidazole modification at C-5 of uridine that is part of tandem G-U wobble base pairs causes slight reduction of thermal stability (DeltaDeltaG(0)(310) < 0.4 kcal mol(-1)) and relatively small change in hydration of short RNA helices. PMID- 16307144 TI - Prodigiosin is a chloride carrier that can function as an anion exchanger. AB - The natural product prodigiosin 1, often described as an H+/Cl- symport cotransporter, can transport Cl- across lipid vesicles via an anion exchange (or antiport) mechanism. PMID- 16307146 TI - A novel sheet 4f-3d mixed-metal pyridine dicarboxylate: synthesis, structure, photophysical properties and its transformation to a perovskite oxide. AB - The synthesis, characterization and photophysical properties of a 4f-3d mixed metal compound, Gd(H2O)3Co[C5N1H3(COO)2]3, are described; the structure is unique, consisting of sheets with large pores (ca. 7 Angstroms diameter) in the sheets and transforms to a perovskite oxide at moderate temperatures. PMID- 16307145 TI - Unique structural topology and reactivities of the ABD tricycle in phomactin A. AB - Stereoselective and transannular reactivities are described for the ABD tricyclic manifold of phomactin A that possesses a unique structural topology. PMID- 16307147 TI - Transformation of nanoporous oxoselenoantimonates into Sb2O3--nanoribbons and nanorods. AB - We have isolated flexible Sb2O3 nanoribbons and nanorods as the main product from the disintegration of nanoporous oxoselenoantimonates of the cetineite type, the size of the one-dimensional nanomaterials obtained ranging up to 15 microm in length with diameters between 8 and 50 nm. PMID- 16307148 TI - Cyclic sulfamidates as lactam precursors. An efficient asymmetric synthesis of ( )-aphanorphine. AB - A short and efficient enantioselective synthesis of (-)-aphanorphine is described based on the use of a cyclic sulfamidate to provide a suitably functionalised lactam that allows for construction of the tricyclic 3-benzazepine scaffold. PMID- 16307149 TI - Pyridine-functionalised ambidextrous gelators: towards catalytic gels. AB - New pyridine containing gelators have been used to prepare Pd-functionalised materials with catalytic activity. PMID- 16307150 TI - Dual chirality control of palladium(II) complexes bearing tropos biphenyl diamine ligands. AB - Axial and center chirality of Pd complexes with tropos biphenyl secondary diamine ligands is shown to be controlled by chiral amide (R)-DABNTf, which can efficiently discriminate between two enantiomeric Pd complexes. PMID- 16307151 TI - Complex PbTe hopper (skeletal) crystals with high hierarchy. AB - A facile and mild solution method has been discovered for the synthesis of complex PbTe hopper crystals in large quantities, which are highly similar to the cubic halite skeletal crystals formed from extreme supersaturation in salt lakes existing in nature. This route may provide a new approach to growing other complex semiconductor structures of high hierarchy. PMID- 16307152 TI - Direct visualisation, by aberration-corrected electron microscopy, of the crystallisation of bimetallic nanoparticle catalysts. AB - Using a scanning transmission electron microscope, corrected for aberration, ultra-high resolution images of the internal structure of nanoparticle clusters, as well as the number of atoms present per cluster, may be directly determined: it is shown that individual bimetallic clusters of Ru10Pt2 have a "molecular" structure and that when they coalesce into larger entities containing ca. 200 atoms they adopt the regular crystalline, and facetted, state of a bulk metal. PMID- 16307153 TI - Selective oxidative para C-C dimerization of 2,6-dimethylphenol. AB - Mechanistic investigations on the oxidative coupling of 2,6-dimethylphenol have led to the development of a selective and efficient procedure to prepare 3,5,3',5'-tetramethyl-biphenyl-4,4'-diol, via a C-C coupling, mediated by a hypervalent form of iodine, i.e. (diacetoxyiodo)benzene and for which a mechanism is proposed. PMID- 16307154 TI - ASCOT-BPLA signals changes to hypertension guidelines. PMID- 16307155 TI - The contribution of diabetes mellitus per se to the severity of coronary artery disease. AB - BACKGROUND: Coronary artery disease (CAD) is the most common chronic, life threatening illness in many Western countries. The risk factors associated with diabetes mellitus and the metabolic syndrome in diabetics are generally considered more important in the evolution of CAD than the diabetic state itself. AIM: This study was undertaken to determine the contribution of diabetes mellitus to the characteristics and severity of coronary artery disease among young patients. METHODS: Thirty diabetics (group 1) were selected from a group of young patients ( or= 6.20 mmol/l. In hypertensive patients, TC (r = 0.24; p < 0.01) and LDL-C (r = 0.20; p < 0.05) were positively correlated to plasma fibrinogen. A positive correlation was also observed between TC and LDL-C (r = 0.91; p < 0.001), HDL-C and CrCl (r = 0.28; p < 0.001), and TG and glucose (r = 0.24; p < 0.01), whereas TG were negatively correlated to HDL-C (r = -0.38; p < 0.001). In multiple linear-regression analysis, LDL-C, HDL-C, TG and age accounted for 87% (R2) of variation in TC levels; plasma glucose and HDL-C for 19% (R2) of variation in TG levels; and TG and CrCl for only 17% (R2) of variation in HDL-C levels. CONCLUSIONS: The present data indicate that dyslipidaemia affects a substantial proportion of healthy and hypertensive Congolese subjects. Furthermore, hypertension is associated with a cluster of risk factors characteristic of the metabolic syndrome, of which overweight/central obesity could be the cornerstone. Management of arterial hypertension should therefore focus both on lowering high blood pressure and correcting associated lipid disorders. PMID- 16307157 TI - Effect of resistance training on cardiorespiratory endurance and coronary artery disease risk. AB - Coronary artery disease (CAD) represents a major medical problem in Western society and is a considerable cause of morbidity and mortality in South Africa. In recent years, epidemiologists have made extensive efforts to define the most common risk factors for CAD and propose preventative measures to limit the spread of the disease. Despite the increasing realisation of the importance of resistance training, the literature has focused primarily on aerobic modes of exercise and their effects on risk for CAD. The aim of this study was to determine whether resistance training could alter cardio-respiratory endurance (VO2max), and thus reduce CAD risk. A quantitative, experimental, comparative research design incorporating a pre-test, a treatment period and a post-test was used. Twenty-eight untrained male volunteers were age matched (mean age: 28 years and seven months) and randomly assigned to either a nonexercising control group (n = 15) or a resistance-training group (n = 13). The study demonstrated no statistically significant change in VO2max for the control group from their pre test (25.097 ml/kg/min) to their post-test (23.778 ml/kg/min) (p = 0.201). However, resistance training significantly (p or=60%, presence of single-level anterior fusion, a high preoperative neurological score, normal epidural SCEPs, and clinical features of brachialgia and cord type. In patients with OPLL, multivariate analysis showed that the long term outcome was positively influenced, in order of importance, by the presence of mixed or localized OPLL, normal epidural SCEPs, high preoperative neurological score, a single-vertebra spondylectomy with anterior fusion, laminoplasty, widening of the transverse area of the cord >or=40%, and an expansion rate of the spinal canal after laminoplasty >or=40%. CONCLUSIONS: We suggest that multivariate analysis is useful for assessing the neurosurgical outcome in patients with cervical compressive myelopathy. PMID- 16307182 TI - Natural course of the minimally symptomatic nonoperated hip in patients with bilateral hip dysplasia treated with contralateral rotational acetabular osteotomy. AB - BACKGROUND: The purpose of this study was to investigate the natural course of the minimally symptomatic nonoperated hip in patients with pre-osteoarthritis or early osteoarthritis in bilaterally dysplastic hips. METHODS: The material consisted of 61 patients with bilaterally dysplastic hips who underwent rotational acetabular osteotomy in their symptomatic hip and nonoperative treatment in their asymptomatic or minimally symptomatic hip. The average follow up was 10.1 years (range 7.0-15.6 years), and the average age at the time of surgery was 38.2 years (range 20-58 years). All patients were divided into two groups by joint congruity of the nonoperated hip at surgery with rotational acetabular osteotomy. RESULTS: On radiographic assessment, 1 of the 35 pre osteoarthritis hips had developed early osteoarthritis, and 6 of the 26 early osteoarthritis hips had progressed to advanced osteoarthritis. The Kaplan-Meier survivorship analysis for the whole group of nonoperated hips, with radiographic progression of osteoarthritis as the endpoint, predicted a 10-year survival rate of 83.7% (95% confidence interval 70%-98%). Significant differences were observed in the radiographic stage (pre- and early osteoarthritis group, P=0.015) and joint congruity (good and fair group, P=0.005). CONCLUSIONS: If the contralateral nonoperated hip has good joint congruity, minimal symptoms, and no or little radiographic change in patients with pre- or early osteoarthritis and bilateral acetabular dysplasia, the probability of radiographic progression is low. PMID- 16307183 TI - Osteoblast lineage properties in giant cell tumors of bone. AB - BACKGROUND: Giant cell tumors of bone (GCTs), among the most common primary bone tumors, are characterized by the formation of abundant osteoclast-like multinucleated giant cells (MNCs). It is not yet clear about the origin of GCTs and which cells in the lesion are the true neoplastic component. Several recent reports suggested that MNCs are osteoclasts induced by stroma-like tumor cells expressing the ligand for receptor activator of NF-kappaB (RANKL), which is a membrane-bound osteoclast differentiation factor. This hypothesis suggests an osteoblast lineage origin of GCTs, although it has long been speculated about GCTs being of mesenchymal stem cell (MSC) origin. METHODS: We investigated the expression of osteoblastic differentiation markers in 10 human GCTs by reverse transcription-polymerase chain reaction and immunohistochemistry. We also performed osteoblastic and adipogenic differentiation assays using cultured cells derived from surgically resected lesions to estimate the stem cell-like properties. RESULTS: GCTs and derived stromal cells expressed many osteoblast lineage marker genes, such as collagen type I, bone sialoprotein, core binding factor a-1, and osteocalcin. Instead of stable expression of mRNA, osteocalcin was not detected among the proteins. The tumor-derived cultures showed osteoblastic but not adipogenic differentiation capability. These findings strongly suggest that GCTs are of osteoblast lineage origin. CONCLUSIONS: Our results indicated that GCTs expressed many osteoblastic markers and showed properties of pre-osteoblast-like cells rather than those of MSCs. These observations may provide some insight into the mechanisms of disease progression and the origin of GCTs. PMID- 16307184 TI - Immunohistological localization of Notch receptors and their ligands Delta and Jagged in synovial tissues of rheumatoid arthritis. AB - BACKGROUND: The interaction of Notch receptors with their transmembrane ligands Delta and Jagged plays an important role not only in the organization of a variety of tissues but also in several genetic disorders and cancer development. The functional involvement of the Notch signaling in rheumatoid arthritis (RA) has been reported previously, but the expression profile of Notch-related molecules, as well as their relation with clinicopathological parameters, remains unclear. METHODS: In this study, we analyzed the immunohistochemical staining pattern of four Notch receptors (Notch1-4) and their ligands (Delta1 and Jagged1) in 14 synovial tissues obtained from 14 RA patients. RESULTS: Notch2 and Notch4 were expressed in limited areas in a few samples or in small blood vessels, respectively. Notch1, Notch3, Delta1, and Jagged1 were overexpressed in the synovial lining and sublining cells on synovial hyperplastic lesions in all samples. Notch1 expression was also observed in T and B lymphocytes of lymphoid follicles independently. Notch1 and Notch3 expression overlapped with that of Jagged1, as determined by confocal microscopy. Activation of Notch1 signaling in the RA synovium was identified using a specific antibody to the cleaved form of Notch1. The expression of these molecules did not show any correlation with clinicopathological parameters. CONCLUSIONS: Our results suggest that Notch signaling is activated in RA synovium but does not necessarily reflect the pathological condition of RA. PMID- 16307185 TI - Type II collagen synthesis in the articular cartilage of a rabbit model of osteoarthritis: expression of type II collagen C-propeptide and mRNA especially during early-stage osteoarthritis. AB - BACKGROUND: The aim of this study was to observe time course changes in type II collagen synthesis in various regions of articular cartilage affected with osteoarthritis (OA) by examining the expression of type II collagen C-propeptide (pCOL II-C) and mRNA in a rabbit OA model. METHODS: Osteoarthritis was experimentally induced by partial lateral meniscectomy in the knees of Japanese white rabbits. The cartilage of the animals was then examined histologically over time. The degenerative area of articular cartilage was divided into three areas, according to the degree of degeneration. The ability to synthesize type II collagen was estimated by the immunohistological staining of pCOL II-C and the in situ hybridization of mRNA in type II collagen. RESULTS: The positive rate of pCOL II-C immunostaining in chondrocytes was highest in the central-degenerative region 1 week after surgery, and the highest rate in the para-degenerative region was observed 2 and 4 weeks after surgery. The percentage of pCOL II-C positive cells increased as the histological degeneration score increased to moderate degeneration and then decreased with further progression of the severity of cartilage degeneration. Examination by in situ hybridization revealed that the regions marked by strong pCOL II-C mRNA expression were similar to those indicated by the immunohistology results. CONCLUSIONS: These results suggest that the type II collagen-synthesizing potential of chondrocytes is highest in moderately degenerated areas of OA articular cartilage. Cartilage repair continues to be seen even as OA advances, although the reaction varies depending on the stage of OA. PMID- 16307186 TI - Down-regulation of basic fibroblast growth factor production from cartilage by excessive mechanical stress. AB - BACKGROUND: We examined the effect of excessive stress on the production of basic fibroblast growth factor (bFGF) by bovine cartilage. METHODS: Bovine articular carpal bone was harvested and cut into 1 cm thick slices horizontally. Continuous compressive stress (0, 2, 20 MPa) was loaded on the cartilage of the carpal bone using a stainless-steel cylinder with a diameter of 8 mm for 1 h. The slices were cultured immediately after compression, and the supernatant of the culture medium was collected for bFGF and nitric oxide (NO) measurement. Interleukin-4 (IL-4) or N(G)-monoethyl-L: -arginine (L: -NMEA) was added to the culture medium in some experiments. RESULTS: Basic FGF production was significantly increased after 36 h of cultivation without mechanical stress (0.40 +/- 0.03 microg/ml). In contrast, the bFGF concentration was not increased by compressive stress of 20 MPa after 36 h of cultivation. An NO inhibitor, L: -NMEA, did not alter the effect of compressive stress on the production of bFGF. IL-4 reduced the production of bFGF by cartilage with or without mechanical stress. CONCLUSIONS: Excessive stress on cartilage inhibits the production of bFGF in an NO-independent manner, and IL-4 plays an important role in the reduction of bFGF. PMID- 16307187 TI - Induction of tumor necrosis factor-alpha in Schwann cells after gradual elongation of rat sciatic nerve. AB - BACKGROUND: Although limb lengthening has become a common treatment, the biochemical responses underlying the adaptation of elongated nerves are unclear. The purpose of this study was to clarify whether expression of cytokines and neurotrophins is altered in gradually elongated peripheral nerves. METHODS: Left sciatic nerves of adult rats were elongated by lengthening the femur up to 20 mm at a rate of 1, 2, or 20 mm/day. The ipsilateral and contralateral sciatic nerves of each group were resected 1, 4, 8, and 16 days after 20 mm of lengthening. mRNAs for interleukin-1beta, interleukin-6, tumor necrosis factor-alpha (TNFalpha), nerve growth factor, brain-derived neurotrophic factor, neurotrophin 3, and neurotrophin-4/5 were semiquantified by reverse transcription-polymerase chain reaction. Histological changes were assessed by immunoperoxidase and immunofluorescence staining. RESULTS: Expression of TNFalpha mRNA was markedly induced in the ipsilateral sciatic nerves of the gradually elongated, 1 mm/day and 2 mm/day groups, although to a lesser extent than in the acutely elongated, 20 mm/day group. In contrast, mRNAs for other factors remained undetectable. The mRNA level for TNFalpha in each group was highest 1 day after 20 mm of lengthening. The highly up-regulated level in the acute group declined rapidly within 4 days and slowly thereafter; in contrast, the decrease in the gradual groups was always slow. Even 16 days later, the levels in all groups remained significantly elevated. Unexpectedly, TNFalpha mRNA expression was also induced in the contralateral side of all groups. Immunohistochemical staining showed that TNFalpha-immunoreactive cells in gradually elongated nerves were also positive for S-100 protein but negative for proliferating nuclear cell antigen, indicating that TNFalpha was produced by nonproliferating Schwann cells. CONCLUSIONS: Gradual nerve elongation by limb lengthening induces production of TNFalpha in Schwann cells. Presumably, TNFalpha plays a critical role in the adaptation of peripheral nerves to elongation. PMID- 16307188 TI - High dosage treatment of nitrogen-containing bisphosphonate ibandronate is required for osseointegration of cementless metal implants. AB - BACKGROUND: There is evidence that application of a bisphosphonate can improve fixation of cementless metal implants by enhancing the extent of osseointegration, but the required dose regimen is still under discussion. The current preclinical study was designed to determine the optimal treatment dose of the nitrogen-containing bisphosphonate ibandronate to improve osseointegration of cementless metal implants. METHODS: The study was conducted in 52 female Sprague Dawley rats in which uncoated and hydroxyapatite-coated titanium implants were surgically inserted into the medullary canal of each femur. The animals were randomly assigned to receive subcutaneous treatment with ibandronate 1 microg/kg body weight (osteoporosis dose) or 25 microg/kg (tumor dose) per day or saline solution for control. RESULTS: Histomorphometric evaluation revealed a significant enhanced extent of osseointegrated implant surface in the high-dose treatment group for both implants compared to the low-dose group and the control group. No significant differences were observed between the two implants in any group. CONCLUSIONS: The results of the present study indicate that improved osseointegration of hydroxyapatite-coated and uncoated titanium implants is dose dependent and requires high-dose application of bisphosphonate ibandronate equivalent to that needed to treat patients with tumor disease. Lower doses equivalent to those for treatment of osteoporosis showed no beneficial effect. PMID- 16307189 TI - Distraction osteogenesis after irradiation in a rabbit model. AB - BACKGROUND: The present study was performed to investigate the effects of preoperative irradiation on distraction osteogenesis, as little is known about how preoperative irradiation delays distraction osteogenesis. METHODS: A single dose of irradiation was applied to the right rear legs of rabbits. This was followed by tibial lengthening at a rate of 0.5 mm/day, which was continued for 4 weeks. Bone regeneration was examined radiographically and histologically. RESULTS: In the irradiation group, the radiographs showed little regeneration during the elongation phase. During the maturation phase, the callus appeared slowly, and its formation was spotty. Furthermore, regeneration was not completed until the fourth week of the maturation period. Histological examination at the end of distraction showed a gap in the distraction consisting of loose connective tissue, with part of the fibrous tissue oriented longitudinally. Four weeks after completion of distraction, the major part of the radiolucent region consisted of cartilage. The spotty osteogenesis was identified as enchondral ossification. Immunohistochemical examination of the regeneration area revealed that the blood vessels were extremely localized, and that the level of expression of vascular endothelial growth factor (VEGF) in the osteoblasts was high. Microangiography showed that vascularization at the distracted sites was poor. Distraction osteogenesis was decreased markedly by preoperative irradiation in terms of both rate and process. The results suggested that most of the osteoprogenitor cells were damaged immediately after irradiation. The high level of VEGF in the osteoblasts and the enchondral ossification also suggested a hypoxic state in the distracted region. CONCLUSIONS: Preoperative irradiation interferes with distraction osteogenesis by inducing a state of poor angiogenesis. PMID- 16307190 TI - Thoracic outlet syndrome combined with carpal tunnel syndrome and Guyon canal syndrome in a child. PMID- 16307191 TI - Bruck syndrome: congenital joint contractures with bone fragility. PMID- 16307192 TI - Lateral meniscus interposition associated with an acute fracture-dislocation of the knee joint in a mogul skier. PMID- 16307193 TI - Alveolar soft part sarcoma of bone. PMID- 16307194 TI - Total knee arthroplasty for gonarthrosis with patellar dislocation. PMID- 16307195 TI - Double-layered lateral meniscus. PMID- 16307196 TI - Surgical treatment for adamantinoma arising from the tibia. PMID- 16307197 TI - Thoracolumbar injury classification and severity score: a new paradigm for the treatment of thoracolumbar spine trauma. AB - BACKGROUND: Contemporary understanding of the biomechanics, natural history, and methods of treating thoracolumbar spine injuries continues to evolve. Current classification schemes of these injuries, however, can be either too simplified or overly complex for clinical use. METHODS: The Spine Trauma Group was given a survey to identify similarities in treatment algorithms for common thoracolumbar injuries, as well as to identify characteristics of injury that played a key role in the decision-making process. RESULTS: Based on the survey, the Spine Trauma Group has developed a classification system and an injury severity score (thoracolumbar injury classification and severity score, or TLICS), which may facilitate communication between physicians and serve as a guideline for treating these injuries. The classification system is based on the morphology of the injury, integrity of the posterior ligamentous complex, and neurological status of the patient. Points are assigned for each category, and the final total points suggest a possible treatment option. CONCLUSIONS: The usefulness of this new system will have to be proven in future studies investigating inter- and intraobserver reliability, as well as long-term outcome studies for operative and nonoperative treatment methods. PMID- 16307198 TI - The morphometric study of the sacrospinal and sacrotuberal ligaments correlated with the morphometry of the pelvis. AB - Perineal neuralgias may be considered as nerve entrapment syndromes, which have been anatomically poorly studied. The pudendal nerve could be compressed between the sacrospinal and the sacrotuberal ligaments. This study tries to find a correlation between the pelvis and the lumbosacral junction morphometry, and sacrospinal and sacrotuberal ligaments morphometry. We did an anatomical study of eight anatomic specimens, and we performed the measures by using the computed tomography scanner. No correlations were found. PMID- 16307199 TI - The human cerebral cortex on MRI: value of the coronal plane. AB - The evaluation of different cortical areas of the cerebral cortex has been analyzed using MRI of 50 normal subjects without any neurological symptoms. This analysis has been made with different spin echo and gradient echo in T1 or T2 in three different planes: horizontal, sagittal and coronal. The most accurate plane to define important cortical areas such as Broca area, Wernicke area, temporal cortex at the level of the superior temporal sulcus, angular gyrus, supra marginal gyrus, hippocampal and parahippocampal cortices as well as that of the parieto or temporo-occipital areas is the coronal plane. Evidently it must be correlated with the other orthogonal planes. To be compared with the main Atlas of Neuroanatomy these sections must be perpendicular or parallel to the plane passing through the anterior and posterior commissures. MRI of patients with neurological disorders must have, as a routine, a series of MR sections performed in the coronal plane, as well as in horizontal and sagittal ones. The coronal plane is certainly the most precise to evaluate these areas involved in language, memory, visuo spatial or behavioral functions. It must be always compared with the rest of the neuroradiological examination and correlated with the clinical neurological signs. PMID- 16307200 TI - Social functioning of young adult intercountry adoptees compared to nonadoptees. AB - Using data from a large adoption and general population cohort, we compared the social functioning of 24- to 30-year-old intercountry adoptees with that of same aged nonadoptees in The Netherlands. Adoptees, compared to nonadoptees, were less likely to have intimate relationships, to live with a partner, and to be married. However, adoptees were not more impaired in their social contacts than nonadoptees. The educational and professional attainment of adoptees was at par with that of the general population. Adopted males showed somewhat less favorable outcomes than adopted females. PMID- 16307201 TI - Metabolite changes in early relapsing-remitting multiple sclerosis. A two year follow-up study. AB - Previous in vivo proton magnetic resonance spectroscopic imaging ((1)H-MRSI) studies have found reduced levels of N-acetyl-aspartate (NAA) in multiple sclerosis (MS) lesions, the surrounding normal-appearing white matter (NAWM) and cortical grey matter (CGM), suggesting neuronal and axonal dysfunction and loss. Other metabolites, such as myoinositol (Ins), creatine (Cr), choline (Cho), and glutamate plus glutamine (Glx), can also be quantified by (1)H-MRSI, and studies have indicated that concentrations of these metabolites may also be altered in MS. Relatively little is known about the time course of such metabolite changes. This preliminary study aimed to characterise changes in total NAA (tNAA, the sum of NAA and N-acetyl-aspartyl-glutamate), Cr, Cho, Ins and Glx concentrations in NAWM and in CGM, and their relationship with clinical outcome, in subjects with clinically early relapsing-remitting MS (RRMS). Twenty RRMS subjects and 10 healthy control subjects underwent (1)H-MRSI examinations yearly for two years. Using the LCModel, tNAA, Cr, Cho, Ins and Glx concentrations were estimated both in NAWM and CGM. At baseline, the concentration of tNAA was significantly reduced in the NAWM of the MS patients compared to the control group (-7%, p = 0.003), as well as in the CGM (-8.7%, p = 0.009). NAWM tNAA concentrations tended to recover from baseline, but otherwise tissue metabolite profiles did not significantly change in the MS subjects, or relatively between MS and healthy control subjects. While neuronal and axonal damage is apparent from the early clinical stages of MS, this study suggests that initially it may be partly reversible. Compared with other MR imaging measures, serial (1)H-MRSI may be relatively less sensitive to progressive pathological tissue changes in early RRMS. PMID- 16307202 TI - Dexamethasone and prognostic factors in adults with bacterial meningitis. AB - OBJECTIVE: To determine the effect of dexamethasone on prognostic factors in adults with bacterial meningitis. DESIGN AND SETTING: A post hoc multivariate analysis of the European Dexamethasone Study. PATIENTS AND PARTICIPANTS: Dexamethasone-treated patients in the European Dexamethasone Study. As internal validation we performed an identical analysis on patients in the placebo group. MEASUREMENTS AND RESULTS: Only focal cerebral abnormalities on admission were predictive for unfavourable outcome in patients treated with early adjunctive dexamethasone (Odds ratio 3.22; 95% confidence interval 1.11-9.35; P=0.03). Other potential prognostic factors failed to achieve statistical significance. An analysis on patients in the placebo group showed prognostic factors comparable with those found in the literature without routine use of dexamethasone. CONCLUSIONS: Routine use of dexamethasone therapy may lead towards new risk stratification in adults with bacterial meningitis. PMID- 16307203 TI - Negative ultrasound findings in patients with cervical artery dissection. Negative ultrasound in CAD. AB - BACKGROUND: Cervical artery dissection (CAD) is a common cause of ischemic stroke in the younger age group. Modern imaging techniques allow the depiction of the mural hematoma, even in CADs with only subtle vessel alterations. The aim of this retrospective study was (1) to characterize the angiological features in CAD and (2) to determine the frequency of initially normal ultrasonography (US) findings. METHODS: 86 patients aged 44 +/- 11 years with CAD of the internal carotid (ICA), (n = 55) or the vertebral artery (VA), (n = 31), admitted to our hospital within 8 days (mean 1.6 days) of symptom onset, were included. CAD was confirmed either by CT-angiography, MRI of the neck, MR-angiography or digital substraction angiography (DSA) and was compared with the results of the initial as well as repeated US examinations of the arteries supplying the brain. RESULTS: In 75 patients (81.2 %) signs of vessel stenosis or occlusion were found while 11 patients (12.8%) with CAD of the ICA (n = 9) and the VA (n = 2) had normal US findings. The site of dissection in the US negative patients was highly variable without a predilection site. In 2 of 7 patients with repeated US examinations, complete vessel occlusion was found on follow-up, while in 5 patients again normal results were found. In four patients, there were changing findings in two alternative confirming imaging methods (MRI/DSA, CT/MRI) and in one patient conflicting findings (CT/MRI). Brain infarctions had occurred in 7 of the initially sonographically normal patients while the other 4 had suffered from transient (n = 2) or local (n = 2) symptoms only. CONCLUSION: Approximately 1 out of 8 patients with subsequently proven CAD has negative initial neurovascular US findings despite comprehensive examination. In patients with suspected CAD and negative US examination, repeated US examinations and further diagnostic imaging, especially MRI is necessary. PMID- 16307204 TI - Corticosteroid use and risk of aneurysmal subarachnoid haemorrhage. AB - OBJECTIVES: Corticosteroids can induce hypertension and inhibit collagen synthesis in the blood vessel wall. Deficiencies in collagen have been found in intracranial aneurysms. Therefore use of corticosteroids could be a risk factor for intracranial aneurysms and aneurysmal subarachnoid haemorrhage (SAH). We investigated the relationship between the systemic use of corticosteroids in the past and the occurrence of aneurysmal SAH. METHODS: We compared the systemic use of corticosteroids (oral or intravenous) in the past between a consecutive series of 1158 patients with aneurysmal SAH and a control group consisting of 1019 patients diagnosed with a primary central nervous system (CNS) tumour. We discriminated between definite use of corticosteroids defined as use mentioned in the medical record and possible use defined as note in the medical record of a disease that may be treated with corticosteroids. We calculated odds ratios (OR) with corresponding 95% confidence intervals (CI) and adjusted for age and sex by means of logistic regression analyses. RESULTS: Twenty (1.7%, 95% CI 1.1-2.7) of the SAH patients and eight (0.8%, 95% CI 0.3-1.5) of the controls had used systemic corticosteroids (OR: 2.22; 95% CI 0.97-5.05; p-value 0.058; adjusted OR 2.23; 95 % CI 0.97-5.15; p-value 0.059). For definite plus possible use the OR was 1.67 (95% CI 1.09-2.54; p-value 0.016) and the adjusted OR 1.52 (95% CI 0.99 2.33; p-value 0.055). CONCLUSIONS: Patients with aneurysmal SAH more often have used systemic corticosteroids in the past than controls. This may suggest that the use of corticosteroids is a risk factor for aneurysmal SAH. PMID- 16307205 TI - A comment on the "correction" of tissue-plasma partition coefficients for drug extraction in eliminating organs by Sven Bjorkman. PMID- 16307206 TI - A dynamical systems analysis of the indirect response model with special emphasis on time to peak response. AB - In this paper we present a mathematical analysis of the four classical indirect response models. We focus on characteristics such as the evolution of the response R(t) with time t, the time of maximal/minimal response T(max) and the area between the response and the baseline AUC(R), and the way these quantities depend on the drug dose, the dynamic parameters such as E(max) and EC50 and the ratio of the fractional turnover rate k(out) to the elimination rate constant k of drug in plasma. We find that depending on the model and on the drug mechanism function, T(max) may increase, decrease, decrease and then increase, or stay the same, as the drug dose is increased. This has important implications for using the shift in T(max) as a diagnostic tool in the selection of an appropriate model. PMID- 16307207 TI - Cluster analysis: an alternative method for covariate selection in population pharmacokinetic modeling. AB - To be analyzed, the heterogeneity characterizing biological data calls for using appropriate models involving numerous variables. A high variable number could become problematic when one needs to determine a priori the most significant variable combination in order to reduce the inter-individual variability (IIV). Alternatively to multiple introductions of single variables, we propose a single introduction of a multivariate variable. We present cluster analysis as a stratification strategy that combines the initial single covariates to build a multivariate categorical covariate. It is an exploratory multivariate analysis that outlines homogeneous categories of individuals (clusters) according to similarities from the set of covariates. It includes many clustering techniques combining a distance measure and a linkage algorithm, and leading to various stratification patterns. The cluster analysis approach is illustrated by a case study on cortisol kinetics in 82 patients after intravenous bolus administration of synacthen (synthetic corticotropin). Using NONMEM, a basic infusion model was initially achieved for cortisol, and then a classical covariate selection was applied to improve IIV. The best fit was between the elimination rate constant k and the body mass index (BMI), which improved IIV of k. An alternative method is presented consisting in the population into homogeneous and non-overlapping groups by applying a cluster analysis. Such categorization (or clustering) was carried out using Euclidean distance and complete-linkage algorithm. This algorithm gave five dissimilar clusters that differed by increasing BMI, obesity duration, and waist-hip ratio. The dispersion of k according to the five clusters showed three distinctvariation ranges a priori, which corresponded a posteriori(after NONMEM modeling) to three sub-populations of k. After grouping the clusters that had similar variation ranges of k, we obtained three final clusters representing non-obese, intermediate, and extreme obese sub-populations. The pharmacokinetic model based on three clusters was better than the basic model, similar to the classical covariate model, but had a stronger interpretability: It showed that the stimulation and elimination of cortisol were higher in the extreme obese followed by intermediate then non-obese subjects. PMID- 16307208 TI - Optimal design for model discrimination and parameter estimation for itraconazole population pharmacokinetics in cystic fibrosis patients. AB - Optimal sampling times are found for a study in which one of the primary purposes is to develop a model of the pharmacokinetics of itraconazole in patients with cystic fibrosis for both capsule and solution doses. The optimal design is expected to produce reliable estimates of population parameters for two different structural PK models. Data collected at these sampling times are also expected to provide the researchers with sufficient information to reasonably discriminate between the two competing structural models. PMID- 16307209 TI - The population pharmacokinetics of citalopram after deliberate self-poisoning: a Bayesian approach. AB - Defining the pharmacokinetics of drugs in overdose is complicated. Deliberate self-poisoning is generally impulsive and associated with poor accuracy in dose history. In addition, early blood samples are rarely collected to characterize the whole plasma-concentration time profile and the effect of decontamination on the pharmacokinetics is uncertain. The aim of this study was to explore a fully Bayesian methodology for population pharmacokinetic analysis of data that arose from deliberate self-poisoning with citalopram. Prior information on the pharmacokinetic parameters was elicited from 14 published studies on citalopram when taken in therapeutic doses. The data set included concentration-time data from 53 patients studied after 63 citalopram overdose events (dose range: 20-1700 mg). Activated charcoal was administered between 0.5 and 4 h after 17 overdose events. The clinical investigator graded the veracity of the patients' dosing history on a 5-point ordinal scale. Inclusion of informative priors stabilised the pharmacokinetic model and the population mean values could be estimated well. There were no indications of non-linear clearance after excessive doses. The final model included an estimated uncertainty of the dose amount which in a simulation study was shown to not affect the model's ability to characterise the effects of activated charcoal. The effect of activated charcoal on clearance and bioavailability was pronounced and resulted in a 72% increase and 22% decrease, respectively. These findings suggest charcoal administration is potentially beneficial after citalopram overdose. The methodology explored seems promising for exploring the dose-exposure relationship in the toxicological settings. PMID- 16307210 TI - A mathematical model for paroxetine antidepressant effect time course and its interaction with pindolol. AB - Although selective 5-HT reuptake inhibitors (SSRIs) block monoamine uptake within hours of administration to patients, their full clinical effect does not appear until 2-4 weeks after treatment onset. Pindolol, a betablocker with weak partial 5-HT1A receptor agonist activity has been shown to produce a more rapid onset of antidepressant action of SSRIs. However, the optimal dosing schedule of pindolol remains controversial. Building on a set-point model described previously for the hypothermic effect of 5-HT agonists, we have developed a model based on the concept of homeostatic control mechanisms, in which SSRIs exert their antidepressant effect by increasing the transduction set-point of the postsynaptic 5-HT1A receptor, and pindolol exerts its effect by increasing the rate of feedback mechanisms. The predictive distribution of the proportion of responders at each day of measurement (based on population simulation from the model) was not significantly different from the proportions observed in two published clinical trials, one with fluoxetine, the other with paroxetine alone or combined with pindolol. The model was applied to the simulation of paroxetine response (clinical score) time course with or without pindolol, after administration of different doses of each drug. The simulated total scores on the MADR scale obtained after treatment with paroxetine alone (20 mg/day) or paroxetine (20 mg/day) with different doses of pindolol (1.5, 7.5 and 37.5 mg/day) support that the reason for inconstant pindolol efficacy is that the 7.5 mg dose is too low. The model might be useful as a basis for clinical trial simulation. PMID- 16307212 TI - Editorial to the first issue dedicated to Lewis Sheiner. PMID- 16307213 TI - Organochlorine insecticides in mudflats of Hong Kong, China. AB - Organochlorine (OC) insecticides were measured in different environmental compartments (including seawater, sediment, and biota) at six sites within the Mai Po Marshes Nature Reserve (MPMNR) and at one site in A Chau (AC) in Hong Kong in June 2002 (wet season) and January 2003 (dry season). MPMNR is a Wetland of International Importance (Ramsar site) situated in the western part of Hong Kong, whereas AC is a relatively remote island in Starling Inlet on the eastern side of the New Territories of Hong Kong. Generally, concentrations of OC insecticides in water, sediment, and biota were greater at MPMNR than at AC. Hexachlorobenzene, dichloro-diphenyl-trichloroethane, and hexachlorocyclohexanes were detected in all samples collected from MPMNR. Preliminary risk assessments were performed by comparing the measured concentrations of OCs in different environmental compartments with the corresponding guidelines promulgated by various environmental protection authorities. Concentrations of some OCs in sediments from certain locations in MPMNR were consistently greater than their corresponding sediment guideline values. These results suggest that the sediment OC concentrations in the sediments of MPMNR might pose a risk to aquatic organisms. Despite this, concentrations of OCs in biota samples were much lower than the relevant guideline values promulgated by government agencies to protect the health of human consumers. It is postulated that the management practices adopted at MPMNR have been effective in limiting the bioaccumulation window (duration) for OCs. This situation may have been aided by a frequent exchange of water between the tidal shrimp ponds at MPMNR and the open sea, again attributable to management practices. The low-cost maintenance practices adopted at MPMNR may potentially be applicable to wetlands in other southeastern Asian countries, thus allowing sustainable fishery production and wildlife conservation. PMID- 16307214 TI - Factors affecting the toxicity of methylmercury injected into eggs. AB - We developed a standardized protocol for comparing the sensitivities of the embryos of different bird species to methylmercury when methylmercury was injected into their eggs. During the course of developing this protocol, we investigated the effects of various factors on the toxicity of the injected methylmercury. Most of our experiments were done with chicken (Gallus domesticus), mallard (Anas platyrhynchos), and ring-necked pheasant (Phasianus colchicus) eggs, all of which were purchased in large numbers from game farms. A smaller amount of work was done with double-crested cormorant (Phalacrocorax auritus) eggs collected from the wild. Several solvents were tested, and corn oil at a rate of 1 microl/g egg contents was selected for the final standardized protocol because it had minimal toxicity to embryos and because methylmercury dissolved in corn oil yielded a dose-response curve in a range of egg concentrations that was similar to the range that causes reproductive impairment when the mother deposits methylmercury into her own eggs. The embryonic stage at which eggs were injected with corn oil altered mercury toxicity; at early stages, the corn oil itself was toxic. Therefore, in the final protocol we standardized the time of injection to occur when each species reached the morphologic equivalent of a 3-day-old chicken embryo. Although solvents can be injected directly into the albumen of an egg, high embryo mortality can occur in the solvent controls because of the formation of air bubbles in the albumen. Our final protocol used corn oil injections into the air cell, which are easier and safer than albumen injections. Most of the methylmercury, when dissolved in corn oil, injected into the air cell passes through the inner shell membrane and into the egg albumen. Most commercial incubators incubate eggs in trays with the air cell end of the egg pointing upward, but we discovered that mercury-induced mortality was too great when eggs were held in this orientation. In addition, some species of bird eggs require incubation on their sides with the eggs being rolled 180 degrees for them to develop normally. Therefore, we adopted a procedure of incubating the eggs of all species on their sides and rolling them 180 degrees every hour. Little has been published about the conditions of temperature, humidity, and the movements to which eggs of wild birds need to be subjected for them to hatch optimally under artificial incubation. Not unexpectedly, hatching success in an artificial incubator is generally less than what natural incubation by the parents can achieve. However, the survival of control embryos of most wild bird species was good (generally > or = 80%) up to within 1 or 2 days of hatching when we incubated the eggs at 37.5 degrees C (or 37.6 degrees C for gallinaceous species) at a relative humidity that resulted in an approximate 15% to 16% loss in egg weight by the end of incubation and by incubating the eggs on their sides and rolling them 180 degrees /h. To improve statistical comparisons, we used survival through 90% of incubation as our measurement to compare survival of controls with survival of eggs injected with graded concentrations of mercury. PMID- 16307215 TI - Toxicity assessment of two soils from Jales mine (Portugal) using plants: growth and biochemical parameters. AB - Contaminants in soils can enter food chains through primary producers. Bioavailable contaminants can induce growth, and reproductive or biochemical changes in plants. To evaluate the bioavailability of heavy metals in two soils from Jales mine surroundings, bioassays with the plants Brassica rapa (RCBr) and Avena sativa were performed. Biochemical parameters (protein and malondialdehyde [MDA] content, and catalase and peroxidase activities) were also measured. The soils had different heavy metal contents: JNC soil contained low heavy metal concentrations, whereas JC soil had high heavy metal contents. Results stressed the difference between species sensitivity, with A. sativa showing no toxicity effects when exposed to both soils. On the other hand, B. rapa presented a decrease in growth parameters when exposed to JNC soil and no changes when exposed to JC soil. A Life Cycle Bioassay confirmed this trend for B. rapa exposed to JNC soil, but also evidenced that JC soil was affecting B. rapa in terms of flower and seed pod production. Biochemical assays showed that plants affected by heavy metals also displayed oxidative stress, with an increase in MDA production, reduction of protein content, and reduction of catalase and peroxidase activities. All bioassays revealed that JNC soil, although with a lower heavy metal content, had a higher bioavailable fraction when compared to JC soil, which consequently increased its toxicity to plants. PMID- 16307216 TI - 2-Deoxy-2-[F-18]fluoro-D-glucose-positron emission tomography in characterization of solid renal masses. AB - The present study was aimed to evaluate the ability of 2-deoxy-2-[F-18]fluoro-D glucose (FDG)-positron emission tomography (PET) in characterization of solid renal masses visualized by computed tomography (CT)/magnetic resonance imaging (MRI) in patients with suspected or known malignancies. METHODS: Twenty-eight solid renal masses (20 unilateral and four bilateral, Size ranges, 1.0-8.4 cm) were evaluated in 24 patients. The results were correlated with histopathology in 15 patients, and clinical follow-up and conventional imaging in all patients. RESULTS: Of the 28 solid renal masses, 10 were primary (nine malignant, one benign) and 18 were metastatic renal tumors. FDG-PET accurately depicted 23 of 27 (85%) malignant renal masses. Of the 10 primary renal tumors, FDG-PET was true positive in eight of nine (89%), true negative in one and false negative in one. The maximum and average standardized uptake values (SUVs) for FDG positive primary renal malignant tumors were 7.9 +/- 4.9 and 6.0 +/- 3.6, respectively. In addition to the characterization of primary tumors, FDG-PET was valuable in primary staging and altered treatment in 30% of patients (three of 10). Of the 18 metastatic renal masses, FDG-PET was positive in 15 (83%) masses. The maximum and average SUVs of metastatic renal masses were 6.1+/- 3.4 and 4.7+/- 2.8, respectively. There was no significant difference in maximum and average SUVs between primary and metastatic renal masses (p=0.3 and p=0.3). CONCLUSION: Despite the physiological excretion of FDG by the kidneys, FDG-PET can be employed effectively in characterization of solid renal masses in patients with suspected or known malignancies. We propose that FDG-PET could be useful as a complimentary modality to conventional imaging in these patients. PMID- 16307217 TI - A heretofore undisclosed crux of eosinophilia-myalgia syndrome: compromised histamine degradation. AB - In contrast to early epidemiological evidence offering links between eosinophilia myalgia syndrome (EMS) and microimpurities of L-tryptophan-containing dietary supplements (LTCDS), this account shows why reliance on a finite impurity from one manufacturer is both unnecessary and insufficient to explain the etiology of EMS. Excessive histamine activity has induced blood eosinophilia and myalgia (Greek: mys, muscle + algos, pain). Termination of the multiple actions of histamine is dependent on particular amine oxidases and histamine-N methyltransferase. Histamine metabolism is rapid when these degradative reactions are operative. The latent effects of incurred histamine can be potentiated and aggravating when these mechanisms are impaired. Overloads of tryptophan supplements cause - among other relevant side-effects - an increased formation of formate and indolyl metabolites, several of which inhibit the degradation of histamine. Moreover, (non-EMS) subjects with hypothalamic-pituitary- adrenal (HPA) axis dysregulation have also manifested greatly increased sensitivities to incurred tryptophan and histamine. A final common pathway for syndromes characterized by eosinophilia with myalgia is now evident. PMID- 16307218 TI - LBP inhibitory peptide reduces endotoxin-induced macrophage activation and mortality. AB - OBJECTIVE AND DESIGN: The aim of this study was to investigate whether P12, a lipopolysaccharide (LPS)-binding protein (LBP) inhibitory peptide could reduce LPS induced inflammation in vitro and in vivo. MATERIAL AND METHODS: Human monocyte-like cell line (U937 cells) was grown in RPMI 1640 and stimulated with PMA in order to induce differentiation to the macrophage stage. A total of 70 Kunming mice (8-12 wk old) were used in our experiments. The effects of P12 on the binding of LPS to U937 cells and alveolar macrophages (AMs) were determined by flow cytometric analysis. Nuclear factor kappa B (NF-kappa B) translocation was evaluated with subunit P65 by Western blotting. The production of tumor necrosis factor-alpha (TNF-alpha), alanine transaminase (ALT), and nitric oxide (NO) as measured by ELISA, enzymatic activity assay, and enzymatic assay with nitrate reductase. Differences among groups were determined using one-way ANOVA test and Fisher exact test. TREATMENT: U937 cells were treated with LPS, LBP, and indicated concentrations of P12. Mice were administered LPS intraperitoneally and P12 via the tail vein. RESULTS: P12 inhibited the binding of FITC-conjugated LPS (FITC-LPS) to U937 cells and AMs. NF-kappa B translocation and the production of TNF-alpha, ALT, and NO induced by LPS was also significantly suppressed by P12. Furthermore P12 protected mice from LPS-induced death. CONCLUSIONS: The results suggest that blockade of LBP at inflammation sites might attenuate LPS-induced circulatory shock. This results in a beneficial effect in a mouse model of endotoxemia. PMID- 16307219 TI - Influence of open versus laparoscopically assisted colectomy on soluble vascular endothelial growth factor (sVEGF) and its soluble receptor 1 (sVEGFR1). AB - INTRODUCTION: Minimal invasive colectomy may attenuate surgery-induced immunomodulation. This may in part be due to a reduced postoperative inflammation mediated angiogenic stimulus directed by the proangiogenic factor VEGF and its neutralizing receptor VEGFR1. Thus, we evaluated perioperative plasma concentrations of soluble VEGF (sVEGF) and soluble VEGFR1 (sVEGFR1) in patients undergoing elective colectomy. METHODS: 60 consecutive patients were randomized to undergo laparoscopically assisted or open right or left sided colectomy. Blood samples were drawn preoperatively, intraoperatively and postoperatively until 30 days after the operation. Commercially available ELISA methods were used for determination of sVEGF and sVEGFR1. RESULTS: Patients with cancer (n = 48) had higher preoperative levels of sVEGF compared to patients with benign disease (n = 12) (p = 0.04), while there was no significant difference in sVEGFR1 levels (p = 0.053). Soluble VEGF (p < 0.0001) and sVEGFR1 (p < 0.0001) levels fluctuated intra- and postoperatively. However, the intra- and postoperative levels of sVEGF and sVEGFR1 were similar at all time points in patients undergoing laparoscopically assisted or open resection. CONCLUSION: Although significant fluctuation in sVEGF and sVEGFR1 concentrations during the perioperative period was shown, patients who underwent laparoscopically assisted resection had similar levels as patients who underwent open resection. PMID- 16307220 TI - Rosiglitazone, an agonist of peroxisome proliferator-activated receptor gamma, reduces pulmonary inflammatory response in a rat model of endotoxemia. AB - OBJECTIVE: The effect of rosiglitazone, a potent peroxisome proliferator activated receptor gamma (PPAR-gamma) agonist, on pulmonary inflammation in endotoxemia was investigated. MATERIALS AND METHODS: Male Wistar rats were given either lipopolysaccharide (LPS, 6 mg/kg i.v.) or saline, pretreated with rosiglitazone (0.3 mg/kg i.v.) or its vehicle (dimethyl sulphoxide) 30 min before LPS. The selective PPAR-gamma antagonist GW9662 (0.3 mg/kg i.v.) was given 20 min before rosiglitazone. Wet/dry weight (W/D) ratio, myeloperoxidase (MPO) activity, malondialdehyde (MDA) as well as TNF-alpha and CINC-1 concentrations were measured in lung tissues 4 h after LPS injection. Expression of ICAM-1, NF-kappaB p65 and PPAR-gamma were also determined by immunohistochemistry or Western blot analysis. RESULTS: Rosiglitazone pretreatment significantly attenuated the increases in W/D ratio, MPO activity and MDA levels, and reduced pulmonary overproduction of TNF-alpha and CINC-1 as well as expression of ICAM-1 following endotoxemia. Rosiglitazone also inhibited the nuclear localization of NF-kappaB and up-regulated the expression of PPAR-gamma protein. The specific PPAR-gamma antagonist GW9662 abolished the effect of rosiglitazone. CONCLUSION: These findings suggest that PPAR-gamma agonists might be used as therapeutic agents in the therapy of inflammatory lung injury related to endotoxemia. PMID- 16307221 TI - Effect and mechanisms of FR167653, a dual inhibitor of TNF-alpha and IL-1, on BCG plus LPS induced-liver injury. AB - OBJECTIVE: To investigate the effect of FR167653, a dual inhibitor of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1), on Bacillus Calmette Guerin (BCG) plus lipopolysaccharide (LPS) induced-liver injury and its mechanisms. MATERIAL AND METHODS: Mouse liver injury was established by tail vein injection of 2.5 mg BCG, and 10 d later with 10 microg LPS. The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were assayed by spectrophotometry. Liver samples were stained with hematoxylin and eosin. Rat hepatocytes (HCs) and Kupffer cells (KCs) were isolated by collagenase IV and pronase perfusion, and purified by density gradient separation. TNF-alpha and IL 1 concentrations were measured with ELISA. TNF-alpha and IL-1 mRNA in KCs was analyzed with RT-PCR. RESULTS: FR167653 significantly decreased the elevated transaminase (ALT, AST) activity in serum of liver injured mice. Meanwhile, the degree of inflammatory cell infiltration and liver cell necrosis was also ameliorated. TNF-alpha and IL-1 production by KCs stimulated with LPS was significantly inhibited by FR167653. RT-PCR analysis demonstrated that FR167653 also reduced the augmented expression of TNF-alpha and IL-1 mRNA in KCs. However, FR167653 up to 10 micromol/L did not have a toxic effect on KC viability. In addition, FR167653 alleviated the HC injury induced by LPS pre-treated Kupffer cell-conditioned medium (KCCM). Addition of anti-IL-1 and anti-TNF-alpha MAbs significantly decreased the ALT level released from HCs incubated with LPS or FR167653 pre-treated KCCM. CONCLUSIONS: TNF-alpha and IL-1 released from activated KCs were involved in BCG plus LPS induced liver injury. FR167653 significantly attenuated hepatocyte injury via inhibition of TNF-alpha and IL-1 released from activated KCs. PMID- 16307222 TI - [History of high-flow priapism: 1960-2005]. AB - High-flow priapism caused by a pathological arterial influx to the cavernous bodies was first described by F.B. Burt in 1960. The pathophysiological differentiation of high- and low-flow priapism was developed in 1983. The development of diagnostic tools for differentiation of different forms of priapism and the progress in the therapy of high-flow priapism from arterial ligation to supraselective embolization is presented. PMID- 16307223 TI - [Joseph Dietl (1804-1878). Reformer of medicine and his contributions to urology]. AB - Two centuries have now passed since the birth of Joseph Dietl, whose uncommon ability, industry, inexhaustible energy, dogged perseverance, and courage contributed to the development of medical science in Europe. He also contributed to reform of the educational system, and to bringing order to the badly neglected city of Cracow. He belonged to the so-called New Vienna School of Medicine. Dietl, chief physician at the Wieden Hospital in Vienna since 1941, had voiced his disbelief in the existing therapy in such strong terms that it was recorded in the history of medicine as the program of therapeutic nihilism. Dietl demonstrated experimentally in Vienna and Cracow that bloodletting in pneumonia is not indicated, since it does not promote recovery and is indeed harmful, raising mortality rates three times compared to those patients who had received dietary treatment only. He eliminated bloodletting in pneumonia therapy. Dietl was the first to investigate and classify Galician and Silesian therapeutic springs as well as work out detailed prescriptions for spa treatments. He understood that treatment in a health resort could not be limited to balneotherapy, and he indicated that climatotherapy and physiotherapy should also play a role. Dietl described the crisis attributable to a kink in the renal vessels or ureter when the kidney dropped. It was also Dietl who gave a description of the floating kidney problem and of kidney partial incarceration, a condition still known as Dietl's crisis. He recommended conservative therapy for these cases. PMID- 16307224 TI - [Testicular metastasis of a metachronous small cell neuroendocrinic prostate cancer after anti-hormonal therapy of a prostatic adenocarcinoma. Case report and literature review]. AB - Secondary malignant testicular tumors are rare, with the exception of infiltrations by lymphomas and leukemias. Metastases are sometimes found in cancers of the lung, gastrointestinal tract, kidney, skin and prostate. Less than about 200 cases of testicular metastasis of prostate cancer have been described in the literature. This is this a very small number in comparison to the high incidence of prostate cancer and the vast number of orchiectomies for operative hormone deprivation in advanced cases. The testicle metastases described are mostly unilateral and are often found as ductal carcinoma of the prostate. In our case, the testicular metastasis from a small cell prostate carcinoma appeared 2.5 years after androgen deprivation of an adenocarcinoma of the prostate. PMID- 16307225 TI - Intestinal apoptotic changes linked to metabolic status in fasted and refed rats. AB - Intestinal apoptosis and expression of apoptosis inducers--the cytokines TNFalpha, TGFbeta1--and the intestinal transcription factor Cdx2, were studied according to two different metabolic and hormonal phases which characterize long term fasting: the long period of protein sparing during which energy expenditure is derived from lipid oxidation (phase II), and the later phase characterized by a rise in body protein utilization and plasma corticosterone (phase III). Apoptosis was further studied in 2, 6, and 24 h refed rats. Morphological apoptotic events were observed by environmental and conventional scanning electron microscopy and a TUNEL test was used to characterize the final stages of apoptotic death. The gene and protein expressions of TNFalpha, TGFbeta1, and Cdx2 were measured. Apoptotic events and TNFalpha, TGFbeta1, and Cdx2 gene and protein expressions did not vary significantly during phase II as compared to the normally fed animals. However, a phase III fasting induced a delay in intestinal epithelial apoptosis, along with a 92, 58, and 25% decrease in TNFalpha, TGFbeta1, and Cdx2 mRNAs, respectively. The amounts of TNFalpha, TGFbeta1, and Cdx2 proteins decreased by 70, 36, and 25%, respectively. Apoptosis was restored rapidly after a 2 h refeeding following the phase III, accompanied by a significant increase in TNFalpha, TGFbeta1, and Cdx2 mRNA and the protein levels, compared to the phase III fasting values. The concomitant decreases in cytokines and Cdx2 and in apoptotic cells during phase III suggest the preservation of enterocytes during this critical fasting period in order to optimize nutrient absorption as soon as food is available and thus, to rapidly restore body mass. PMID- 16307226 TI - Assessment of ancestry probabilities in the presence of genotyping errors. AB - This paper extends an approach for estimating the ancestry probability, the probability that an inbred line is an ancestor of a given hybrid, to account for genotyping errors. The effect of such errors on ancestry probability estimates is evaluated through simulation. The simulation study shows that if misclassification is ignored, then ancestry probabilities may be slightly overestimated. The sensitivity of ancestry probability calculations to the assumed genotyping error rate is also assessed. PMID- 16307227 TI - SSR allelic variation in almond (Prunus dulcis Mill.). AB - Sixteen SSR markers including eight EST-SSR and eight genomic SSRs were used for genetic diversity analysis of 23 Chinese and 15 international almond cultivars. EST- and genomic SSR markers previously reported in species of Prunus, mainly peach, proved to be useful for almond genetic analysis. DNA sequences of 117 alleles of six of the 16 SSR loci were analysed to reveal sequence variation among the 38 almond accessions. For the four SSR loci with AG/CT repeats, no insertions or deletions were observed in the flanking regions of the 98 alleles sequenced. Allelic size variation of these loci resulted exclusively from differences in the structures of repeat motifs, which involved interruptions or occurrences of new motif repeats in addition to varying number of AG/CT repeats. Some alleles had a high number of uninterrupted repeat motifs, indicating that SSR mutational patterns differ among alleles at a given SSR locus within the almond species. Allelic homoplasy was observed in the SSR loci because of base substitutions, interruptions or compound repeat motifs. Substitutions in the repeat regions were found at two SSR loci, suggesting that point mutations operate on SSRs and hinder the further SSR expansion by introducing repeat interruptions to stabilize SSR loci. Furthermore, it was shown that some potential point mutations in the flanking regions are linked with new SSR repeat motif variation in almond and peach. PMID- 16307228 TI - Inheritance and interactions of incompatibility alleles in the tetraploid sour cherry. AB - Three progenies of sour cherry (Prunus cerasus) were analysed to correlate self (in)compatibility status with S-RNase phenotype in this allotetraploid hybrid of sweet and ground cherry. Self-(in)compatibility was assessed in the field and by monitoring pollen tube growth after selfing. The S-RNase phenotypes were determined by isoelectric focusing of stylar proteins and staining for RNase activity and, for the parents, confirmed by PCR. Seedling phenotypes were generally consistent with disomic segregation of S-RNase alleles. The genetic arrangements of the parents were deduced to be 'Koroser' (self-incompatible) S1S4.S(B) S(D), 'Schattenmorelle' (self-compatible) S6S13.S(B)S(B), and clone 43.87 (self-compatible) S4S13.S(B)S(B), where "." separates the two homologous genomes. The presence of S4 and S6 alleles at the same locus led to self incompatibility, whereas S13 and S(B) at homologous loci led to self compatibility. The failure of certain heteroallelic genotypes in the three crosses or in the self-incompatible seedlings indicates that S4 and S6 are dominant to S(B). However, the success of S13S(B) pollen on styles expressing corresponding S-RNases indicates competitive interaction or lack of pollen-S components. In general, the universal compatibility of S13S(B) pollen may explain the frequent occurrence of S13 and S(B) together in sour cherry cultivars. Alleles S(B) and S(D), that are presumed to derive from ground cherry, and S13, presumably from sweet cherry, were sequenced. Our findings contribute to an understanding of inheritance of self-(in)compatibility, facilitate screening of progenies for self-compatibility and provide a basis for studying molecular interactions in heteroallelic pollen. PMID- 16307229 TI - Development of an integrated genetic map of a sugarcane (Saccharum spp.) commercial cross, based on a maximum-likelihood approach for estimation of linkage and linkage phases. AB - Sugarcane (Saccharum spp.) is a clonally propagated outcrossing polyploid crop of great importance in tropical agriculture. Up to now, all sugarcane genetic maps had been developed using either full-sib progenies derived from interspecific crosses or from selfing, both approaches not directly adopted in conventional breeding. We have developed a single integrated genetic map using a population derived from a cross between two pre-commercial cultivars ('SP80-180' x 'SP80 4966') using a novel approach based on the simultaneous maximum-likelihood estimation of linkage and linkage phases method specially designed for outcrossing species. From a total of 1,118 single-dose markers (RFLP, SSR and AFLP) identified, 39% derived from a testcross configuration between the parents segregating in a 1:1 fashion, while 61% segregated 3:1, representing heterozygous markers in both parents with the same genotypes. The markers segregating 3:1 were used to establish linkage between the testcross markers. The final map comprised of 357 linked markers, including 57 RFLPs, 64 SSRs and 236 AFLPs that were assigned to 131 co-segregation groups, considering a LOD score of 5, and a recombination fraction of 37.5 cM with map distances estimated by Kosambi function. The co-segregation groups represented a total map length of 2,602.4 cM, with a marker density of 7.3 cM. When the same data were analyzed using JoinMap software, only 217 linked markers were assigned to 98 co-segregation groups, spanning 1,340 cM, with a marker density of 6.2 cM. The maximum-likelihood approach reduced the number of unlinked markers to 761 (68.0%), compared to 901 (80.5%) using JoinMap. All the co-segregation groups obtained using JoinMap were present in the map constructed based on the maximum-likelihood method. Differences on the marker order within the co-segregation groups were observed between the two maps. Based on RFLP and SSR markers, 42 of the 131 co-segregation groups were assembled into 12 putative homology groups. Overall, the simultaneous maximum-likelihood estimation of linkage and linkage phases was more efficient than the method used by JoinMap to generate an integrated genetic map of sugarcane. PMID- 16307230 TI - QTL analysis of fruit components in the progeny of a Rennell Island Tall coconut (Cocos nucifera L.) individual. AB - We investigated the genetic factors controlling fruit components in coconut by performing QTL analyses for fruit component weights and ratios in a segregating progeny of a Rennell Island Tall genotype. The underlying linkage map of this population was already established in a previous study, as well as QTL analyses for fruit production, which were used to complement our results. The addition of 53 new markers (mainly SSRs) led to minor amendments in the map. A total of 52 putative QTLs were identified for the 11 traits under study. Thirty-four of them were grouped in six small clusters, which probably correspond to single pleiotropic genes. Some additional QTLs located apart from these clusters also had relatively large effects on the individual traits. The QTLs for fruit component weight, endosperm humidity and fruit production were found at different locations in the genome, suggesting that efficient marker-assisted selection for yield can be achieved by selecting QTLs for the individual components. The detected QTLs descend from a genotype belonging to the "Pacific" coconut group. Based on the known molecular and phenotypic differences between "Pacific" and "Indo-Atlantic" coconuts, we suggest that a large fraction of coconut genetic diversity is still to be investigated by studying populations derived from crosses between these groups. PMID- 16307231 TI - Impact of IDDM2 on disease pathogenesis and progression in children with newly diagnosed type 1 diabetes: reduced insulin antibody titres and preserved beta cell function. AB - AIMS/HYPOTHESIS: The insulin-dependent diabetes mellitus 2 gene (IDDM2) is a type 1 diabetes susceptibility locus contributed to by the variable number of tandem repeats (VNTR) upstream of the insulin gene (INS). We investigated the association between INS VNTR class III alleles (-23HphIA/T) and both insulin antibody presentation and residual beta cell function during the first year after diagnosis in 257 children with type 1 diabetes. MATERIALS AND METHODS: To estimate C-peptide levels and autoantibody presentation, patients underwent a meal-stimulated C-peptide test 1, 6, and 12 months after diagnosis. The insulin 23HphIA/T variant was used as a marker of class III alleles and genotyped by PCR RFLP. RESULTS: The insulin antibody titres at 1 and 6 months were significantly lower in the class III/III and class I/III genotype groups than in the class I/I genotype group (p = 0.01). Class III alleles were also associated with residual beta cell function 12 months after diagnosis and independently of age, sex, BMI, insulin antibody titres, and HLA-risk genotype group (p = 0.03). The C-peptide level was twice as high among class III/III genotypes as in class I/I and class I/III genotypes (319 vs 131 and 166 pmol/l, p=0.01). Furthermore, the class III/III genotype had a 1.1% reduction in HbA(1)c after adjustment for insulin dose (p = 0.04). CONCLUSIONS/INTERPRETATION: These findings suggest a direct connection in vivo between INS VNTR class III alleles, a decreased humoral immune response to insulin, and preservation of beta cell function in recent-onset type 1 diabetes. PMID- 16307232 TI - Lack of genotoxic effect in workers exposed to very low doses of 1,3-butadiene. AB - 1,3-Butadiene (BD), a probable carcinogen to humans, has been shown to have an ill-defined genotoxicity in occupationally exposed workers. In the present study, the influence of exposure to very low doses of BD and to cigarette smoking was investigated on some cytogenetic endpoints, namely, sister chromatid exchanges (SCE), chromosomal aberrations (CA) and cells with a high frequency of SCE (HFC), in peripheral blood lymphocytes. Twenty-seven male workers employed in a petrochemical plant and 26 matched controls were included in the study. As regards the airborne BD values, there was a significant difference between exposed (median BD value 1.5, min-max 0.2-69.0 microg/m3) and non-exposed workers (median BD value 0.4, min-max <0.1-3.8 microg/m3). Genotoxic biomarkers were not able to distinguish between the two groups. The frequency of SCE was higher in smokers than in non-smokers (p=0.001), with a positive correlation between the number of cigarettes smoked per day and both SCE (r=0.4; p=0.004) and HFC frequency (r=0.3; p=0.04). Multiple regression analysis confirmed the influence of cigarette smoking on the level of SCE and HFC, while these parameters were not affected by personal exposure to BD. Overall, the biomarkers of genotoxic effect investigated in our study were not able to discriminate between workers with a very low exposure to BD and controls, while it was possible to distinguish between smokers and non-smokers on the basis of SCE. PMID- 16307233 TI - Control of aperture closure during reach-to-grasp movements in Parkinson's disease. AB - This study examined whether the pattern of coordination between arm-reaching toward an object (hand transport) and the initiation of aperture closure for grasping is different between PD patients and healthy individuals, and whether that pattern is affected by the necessity to quickly adjust the reach-to-grasp movement in response to an unexpected shift of target location. Subjects reached for and grasped a vertical dowel, the location of which was indicated by illuminating one of the three dowels placed on a horizontal plane. In control conditions, target location was fixed during the trial. In perturbation conditions, target location was shifted instantaneously by switching the illumination to a different dowel during the reach. The hand distance from the target at which the subject initiated aperture closure (aperture closure distance) was similar for both the control and perturbation conditions within each group of subjects. However, that distance was significantly closer to the target in the PD group than in the control group. The timing of aperture closure initiation varied considerably across the trials in both groups of subjects. In contrast, aperture closure distance was relatively invariant, suggesting that aperture closure initiation was determined by spatial parameters of arm kinematics rather than temporal parameters. The linear regression analysis of aperture closure distance showed that the distance was highly predictable based on the following three parameters: the amplitude of maximum grip aperture, hand velocity, and hand acceleration. This result implies that a control law, the arguments of which include the above parameters, governs the initiation of aperture closure. Further analysis revealed that the control law was very similar between the subject groups under each condition as well as between the control and perturbation conditions for each group. Consequently, the shorter aperture closure distance observed in PD patients apparently is a result of the hypometria of their grip aperture and bradykinesia of hand transport movement, rather than a consequence of a deficit in transport-grasp coordination. It is also concluded that the perturbation of target location does not disrupt the transport-grasp coordination in either healthy individuals or PD patients. PMID- 16307234 TI - Systematic changes in the duration and precision of interception in response to variation of amplitude and effector size. AB - The results of two experiments are reported that examined how performance in a simple interceptive action (hitting a moving target) was influenced by the speed of the target, the size of the intercepting effector and the distance moved to make the interception. In Experiment 1, target speed and the width of the intercepting manipulandum (bat) were varied. The hypothesis that people make briefer movements, when the temporal accuracy and precision demands of the task are high, predicts that bat width and target speed will divisively interact in their effect on movement time (MT) and that shorter MTs will be associated with a smaller temporal variable error (VE). An alternative hypothesis that people initiate movement when the rate of expansion (ROE) of the target's image reaches a specific, fixed criterion value predicts that bat width will have no effect on MT. The results supported the first hypothesis: a statistically reliable interaction of the predicted form was obtained and the temporal VE was smaller for briefer movements. In Experiment 2, distance to move and target speed were varied. MT increased in direct proportion to distance and there was a divisive interaction between distance and speed; as in Experiment 1, temporal VE was smaller for briefer movements. The pattern of results could not be explained by the strategy of initiating movement at a fixed value of the ROE or at a fixed value of any other perceptual variable potentially available for initiating movement. It is argued that the results support pre-programming of MT with movement initiated when the target's time to arrival at the interception location reaches a criterion value that is matched to the pre-programmed MT. The data supported completely open-loop control when MT was less than between 200 and 240 ms with corrective sub-movements increasingly frequent for movements of longer duration. PMID- 16307235 TI - Cue familiarity is represented in monkey medial prefrontal cortex during visuomotor association learning. AB - To examine functional roles of the medial prefrontal cortex (mPFC) in visuomotor association learning, neuronal activity in the mPFC of a behaving monkey was recorded during this learning. The monkey was presented a cueing visual stimulus, and required to push, pull or turn a manipulator according to the cue following a delay period. Under the control condition, three cues (circle, triangle and square) instructed the monkey to the three responses in a block of trials. After 2 months of training the animal was familiar with these cue-response associations. Under the learning condition, two of the three familiar cues and one novel cue were presented in a block. The monkey initially did not know what the novel cue instructed at first and learned a new cue-response association by trial and error. Neurons in the mPFC showed marked responses to cue presentation, and cue responses changed depending on whether cues were familiar or novel. A group of mPFC neurons responded to novel cues, but not to familiar cues. Another group of neurons responded to familiar cues, but not to novel cues. In a subgroup of these familiar cue-selective neurons, cue response was increased under the learning condition compared to the control condition. These results suggest that mPFC neurons differentiate between familiar and novel instructions, and that the neurons responsive to familiar stimuli enhance their modulations when both familiar and novel instructions have to be processed during task performance. PMID- 16307236 TI - The visual control of stepping operates in real time: Evidence from a pictorial illusion. AB - Recent studies have suggested important similarities between the visual control of upper and lower limb actions despite the different effectors, time scales, and regions of space associated with the two types of actions. Drawing on previous findings from upper limb studies, we show here that the effect of a Muller-Lyer illusion on stepping amplitude is greatly increased when the target display is occluded as compared to when it is visible during the reaction time period (i.e., brief delay and 3-s delay conditions versus full vision and open-loop conditions). The present results suggest that the control of stepping is mediated by different systems when the target is visible as compared to when it is occluded immediately before movement initiation; when target vision is not available during the reaction time period, the control of stepping is more highly dependent upon a non-veridical, perceptual representation of the target environment. These findings are consistent with the idea that, much like grasping, vision during the reaction time period is critical for the optimal control of stepping; that is, the visual control of stepping operates in real time. PMID- 16307237 TI - Postcentral neurons with covert receptive fields in conscious macaque monkeys: their selective responsiveness to simultaneous two-point stimuli applied to discrete oral portions. AB - The representation of the oral structures in the postcentral somatosensory cortex was studied in conscious macaque monkeys by recording the activity of single neurons. A total of 2,807 neurons were isolated in the oral regions of three hemispheres in two animals. Of these, 375 neurons (area 3a, 3; area 3b, 123; area 1, 99; area 2, 150) lacked an apparent receptive field (RF), and their relative frequency was significantly higher in area 2 (19%) than in more rostral areas (area 3a, 8%; area 3b, 10%; area 1, 12%). We tested the responsiveness of these neurons to stimuli applied simultaneously to two discrete, but functionally related, oral structures (interstructural two-point stimuli: iTPS). Neurons in areas 3a, 3b, and 1 that lacked an apparent RF were not responsive to iTPS. However, 35 neurons in area 2 responded stably to iTPS applied to either of the following sets of oral structures: the tongue and incisors (n=18), incisors and lip (n=9), lip and tongue (n=12), or upper and lower lips (n=8). Of them, 19 neurons were activated during self-movements such as tongue protrusion, lip licking, and food manipulation. The neurons selectively responsive to iTPS might detect converging inputs from different oral structures and play a pivotal role in detecting objects straddling different oral structures and the mutual contact of oral structures. PMID- 16307238 TI - Control of hand orientation and arm movement during reach and grasp. AB - We studied the coordination of arm and wrist motion in a task requiring fine control of hand orientation. Subjects were instructed to reach and grasp one of two targets positioned in the frontal plane at various orientations. The task was performed under three target conditions: fixed orientation, predictably perturbed orientation, and randomly perturbed orientation. For fixed target orientations, the hand began to rotate to the required orientation from the beginning of the reach. Hand peak supination angles scaled linearly with target orientations. The trajectories of hand/arm joint angles also had a one-to-one relationship with different target orientations. These demonstrate that target orientation is a constraint on the hand/arm final orientation, a control variable to be specified in advance by the central nervous system (CNS). Under perturbation conditions, subjects were still able to complete the task smoothly. In the early trials of the predictable perturbation, the hand rotated first to the original target orientation and then corrected for the final target orientation. Initial corrections occurred about 200 ms after the onset of perturbation. This latency decreased as the subjects adapted to the perturbation, and the hand orientation trajectory shifted to match the unperturbed trajectory for the final orientation. By contrast, we observed no clear changes in orientation trajectory under the randomly perturbed conditions. These suggest that feedback control is important to the execution of the movement, but that the CNS tends to optimize feedforward planning rather than feedback correction when the disturbance information is predictable. PMID- 16307239 TI - Changes in perception of active but not passive turning following stepping on the rotating treadmill. AB - The purpose of this work was to determine whether walking on a rotating disc would cause changes in perception of active and passive turning. Ten healthy control subjects wore a blindfold and earplugs while completing trials of active and passive turning. For active conditions, subjects were told the direction and amplitude of the desired turn and then attempted to turn in place the specified amount via actively stepping. For passive conditions, subjects were told that the disc they stood on would turn and they were to press a button when they had traveled the specified amplitude. Subjects completed active and passive trials to the left and right with amplitudes of 90, 180, 270, and 360 degrees. Subjects then stepped in place for 15 min on a disc rotating clockwise at 90 degrees /s, after which they repeated the trials of active and passive turning. Following rotating treadmill stimulation, subjects asked to turn in the direction opposite disc rotation (i.e., to the left) consistently and significantly overshot their targets in active trials. There were no changes in accuracy of active turning to the right or passive turning in either direction. This indicates that adaptation to the rotating treadmill is expressed even when subjects have a conscious intent to turn. The positive after-effects of the rotating treadmill add to the intended active turning in the direction of the after-effect but there is no decrement in active turning in the direction opposite the after-effect. PMID- 16307240 TI - Transneuronal tracing of vestibulo-trigeminal pathways innervating the masseter muscle in the rat. AB - Previous studies reported that the activity of trigeminal motoneurons innervating masseter muscles is modulated by vestibular inputs. We performed the present study to provide an anatomical substrate for these physiological observations. The transynaptic retrograde tracer pseudorabies virus-Bartha was injected into multiple sites of the lower third of the superficial layer of the masseter muscle in rats, a subset of which underwent a sympathectomy prior to virus injections, and the animals were euthanized 24-120 h later. Labeled masseteric motoneurons were first found in the ipsilateral trigeminal motor nucleus following a 24-h postinoculation period; subsequent to 72-h survival times, the number of infected motoneurons increased, and at > or =96 h many of these cells showed signs of cytopathic changes. Following 72-h survival times, a few transynaptically labeled neurons appeared bilaterally in the medial vestibular nucleus (MVe) and the caudal prepositus hypoglossi (PH) and in the ipsilateral spinal vestibular nucleus (SpVe). At survival times of 96-120 h, labeled neurons were consistently observed bilaterally in all vestibular nuclei (VN), although the highest concentration of infected cells was located in the caudal part of the MVe, the SpVe, and the caudal portion of PH. The distribution and density of labeling in the VN and PH were similar in sympathectomized and nonsympathectomized rats. These anatomical data provide the first direct evidence that neurons in the VN and PH project bilaterally to populations of motoneurons innervating the lower third of the superficial layer of the masseter muscle. The MVe, PH, and SpVe appear to play a predominant integrative role in producing vestibulo-trigeminal responses. PMID- 16307241 TI - Age reduces cortical reciprocal inhibition in humans. AB - Age alters the control of voluntary movement. A widely observed age-related adaptation is the heightened activation of the antagonist muscles during voluntary movements. We examined the possibility that age also modifies cortical reciprocal inhibition. In young (age 27, n = 6) and old (age 73, n = 6) adults a mild conditioning electrical stimulus was delivered to the median nerve at the elbow. The test stimulus, delivered by transcranial magnetic stimulation (TMS) at 1-ms increments between 11 and 24 ms after the electrical conditioning stimulus, evoked motor potentials (MEP) in the extensor carpi radialis (ECR) and flexor carpi radialis (FCR). The absolute TMS intensity, expressed as the percent of stimulator output, used to produce 1-mV control MEPs in the ECR was similar in young (mean 58.5, standard deviation +/-12.8%) and old adults (60.3+/-20.3%, P = 0.855). The size of the control MEP in the ECR was also similar in young (0.98+/ 0.10 mV) and old subjects (0.90+/-0.14 mV, P = 0.686). The age by conditioning interval interaction (P = 0.001) showed that the MEPs in the ECR were significantly depressed at 14, 15, 16, 17, 18, and 19 ms (range 55.5--65.9% of control, all P < 0.05) compared with control value of 100% and with old adults who showed no depression. The MEPs remained at control level in the FCR and were also unaffected in the first dorsal interosseus. These data confirm the existence of cortical reciprocal inhibition reported previously in young humans and show that age reduces this inhibition similarly to the previously reported reduction of spinal reciprocal inhibition reported in old adults. Activation of agonist and antagonist muscle pairs are most likely organized around a dual system of cortically and spinally mediated reciprocal inhibition that is altered by age. The data also indicate the need to use age-matched control subjects when comparing individuals with abnormalities resulting from disorders that occur at an old age. PMID- 16307242 TI - Differential influence of vision and proprioception on control of movement distance. AB - The purpose of this study was to investigate the contribution of proprioceptive and visual information about initial limb position in controlling the distance of rapid, single-joint reaching movements. Using a virtual reality environment, we systematically changed the relationship between actual and visually displayed hand position as subjects' positioned a cursor within a start circle. No visual feedback was given during the movement. Subjects reached two visual targets (115 and 125 degrees elbow angle) from four start locations (90, 95, 100, and 105 degrees elbow angle) under four mismatch conditions (0, 5, 10, or 15 degrees). A 2 x 4 x 4 ANOVA enabled us to ask whether the subjects controlled the movement distance in accord with the virtual, or the actual hand location. Our results indicate that the movement distance was mainly controlled according to the virtual start location. Whereas distance modification was most extensive for the closer target, analysis of acceleration profiles revealed that, regardless of target position, visual information about start location determined the initial peak in tangential hand acceleration. Peak acceleration scaled with peak velocity and movement distance, a phenomenon termed "pulse-height" control. In contrast, proprioceptive information about actual hand location determined the duration of acceleration, which also scaled with peak velocity and movement distance, a phenomenon termed "pulse-width" control. Because pulse-height and pulse-width mechanisms reflect movement planning and sensory-based corrective processes, respectively, our current findings indicate that vision is used primarily for planning movement distance, while proprioception is used primarily for online corrections during rapid, unseen movements toward visual targets. PMID- 16307243 TI - The relation between force and movement when grasping an object with a precision grip. AB - When reaching out for objects, the digits' paths curve so that they approach their positions of contact moving more or less perpendicularly to the local surface orientation. This increases the accuracy of positioning the digits and ensures that any forces exerted at contact are nearly perpendicular to the surface, so that friction will prevent the digits from slipping along the surface. When lifting the object a similar force perpendicular to the surface is needed to prevent the object from slipping from one's fingers. In order to determine whether these two issues are dealt with simultaneously we let subjects pick up a cube from three different starting positions and measured the digits' movements and forces from before contact until the moment the cube started moving. The impact force was low. After impact, the digits spent about 200 ms in contact with the surface of the cube before the latter started to move. The digits first decelerated, and then they gradually built up the grip- and lift forces to move the cube upwards. We found no direct relationship between the control of the reaching movement towards the object and the force applied at the surface of the object to pick it up. We conclude that the reaching and lifting movements are quite independent. PMID- 16307244 TI - Startle responses in Parkinson patients during human gait. AB - Falls frequently occur in patients with Parkinson's disease (Bloem et al. 2001). One potential source for such falls during walking might be caused by the reaction to loud noises. In normal subjects startle reactions are well integrated in the locomotor activity (Nieuwenhuijzen et al. 2000), but whether this is also achieved in Parkinson patients is unknown. Therefore, in the present study, the startle response during walking was studied in eight patients with Parkinson's disease and in eight healthy subjects. To examine how startle reactions are incorporated in an ongoing gait pattern of these patients, unexpected auditory stimuli were presented in six phases of the step cycle during walking on a treadmill. For both legs electromyographic activity was recorded from biceps femoris and tibialis anterior. In addition, we measured the stance and swing phases of both legs, along with the knee angles of both legs and the left ankle angle. In all subjects and all muscles, responses were detected. The pattern of the responses, latency, duration, and phase-dependent modulation was similar in both groups. However, the mean response amplitude was larger in patients due to a smaller habituation rate. No correlation was found between the degree of habituation and disease severity. Moreover, a decreased habituation was already observed in mildly affected patients, indicating that habituation of the startle response is a sensitive measure of Parkinson's disease. The results complement the earlier findings of reduced habituation of blink responses in Parkinson's disease. With respect to behavioral changes in healthy subjects we observed that startle stimuli induced a shortening of the step cycle and a decrease in range of motion. In the patient group, less shortening of the subsequent step cycle and no decrease in range of motion of the knee and ankle was seen. It is argued that the observed changes might contribute to the high incidence of falls in patients with Parkinson's disease. PMID- 16307245 TI - Phoria adaptation after sustained symmetrical convergence: Influence of saccades. AB - We recorded divergence eye movements after short (4 s) and long (36 s) periods of sustained symmetrical convergence (30 degrees) in nine normal human subjects using the search coil technique. Following the long period of convergence, alignment after the initial 1,250 ms of divergence was more converged than after the short period of convergence, showing short-term "phoria adaptation". The first 1,000 ms of divergence, however, could be slower, faster or relatively unchanged, depending upon the subject. A change in the timing and/or amplitude of associated saccades (which accelerate ongoing vergence) between the long and short stimuli accounted for much of the difference in the rate of divergence. The differences in saccade pattern during early divergence following the long and short periods of convergence may reflect changes in attentional focus (to near or to far). PMID- 16307246 TI - Bimanual adaptation: internal representations of bimanual rhythmic movements. AB - From tying your shoes and clipping your tie to the claps at the end of a fine seminar, bimanual coordination plays a major role in our daily activities. An important phenomenon in bimanual coordination is the predisposition toward mirror symmetry in the performance of bimanual rhythmic movements. Although learning and adaptation in bimanual coordination are phenomena that have been observed, they have not been studied in the context of adaptive control and internal representations-approaches that were successfully employed in the arena of reaching movements and adaptation to force perturbations. In this paper we examine the dynamics of the learning mechanisms involved when subjects are trained to perform a bimanual non-harmonic polyrhythm in a bimanual index finger tapping task. Subjects are trained in this task implicitly, using altered visual feedback, while their performance is continuously monitored throughout the experiment. Our experimental results indicate the existence of significant (p<<0.01) learning curves (i.e., error plots with significantly negative slopes) during training and aftereffects with a washout period after the visual feedback ceases to be altered. These results confirm the formation of internal representations in bimanual motor control. We present a simple, physiologically plausible, neural model that combines feedback and adaptation in the control process and which is able to reproduce key phenomena of bimanual coordination and adaptation. PMID- 16307247 TI - Physiology of modulation of motor cortex excitability by low-frequency suprathreshold repetitive transcranial magnetic stimulation. AB - Many studies show consistently that repetitive transcranial magnetic stimulation (rTMS) with a frequency of 1 Hz and an intensity above the resting motor threshold (RMT) performed for several minutes over the primary motor cortex (M1) leads to a depression of cortical excitability. Furthermore, most studies concur on a facilitation of the non-stimulated contralateral M1. Little is known, however, about the physiological mechanisms underlying these effects. In 11 healthy volunteers, we stimulated the left M1 for 15 min with 1 Hz-rTMS of 115% RMT. Before, immediately after, and 30 min after the rTMS train, we examined short-interval intracortical inhibition (SICI; interstimulus interval (ISI) of 2 and 4 ms), intracortical facilitation (ICF; ISI 10 ms), and short-interval intracortical facilitation (SICF; ISI 1.5 ms) with established paired-pulse protocols. Mean unconditioned motor evoked potential (MEP) amplitudes and RMT were also measured. Two sessions were run at least 1 week apart, in one excitability of the stimulated M1 was tested, in the other one excitability of the non-stimulated M1. rTMS led to the expected reduction of MEP amplitude of the stimulated M1, which was significant only immediately after the rTMS train. rTMS increased MEP amplitude of the non-stimulated M1, which lasted for at least 30 min. RMT, SICI, ICF and SICF did not show any significant change in either M1, except for a long lasting increase of SICF in the non-stimulated M1. In conclusion, the MEP increase in the non-stimulated M1 lasted longer than the MEP decrease in the stimulated M1. Only the long-lasting MEP increase was associated with a specific change in intracortical excitability (increase in SICF). Modulation of motor cortical inhibition did not play a role in explaining the rTMS induced changes in MEP amplitude. PMID- 16307248 TI - Eye-position dependence of torsional velocity during step-ramp pursuit and transient yaw rotation in humans. AB - The time course of eye-position-dependent torsion during transient horizontal pursuit and yaw rotation was examined in seven normal human subjects. The stimuli consisted of step-ramp target motion (25, 40 degrees /s) and brief chair rotation (approximately 200 degrees /s(2) accelerated to 40 degrees /s) at three different vertical positions (center 0 degrees , up or down 15 degrees ). Three-dimensional eye movements were recorded with dual search coils. The kinematics of pursuit and the rotational vestibulo-ocular reflex (rVOR) were assessed by determining the tilt-angle slope, a measure of the variation of the axis of eye-velocity with vertical eye position. We found that the tilt-angle slope during pursuit was initially 0.4+/-0.07 (mean+/-95% confidence interval) and then gradually rose to 0.64+/-0.04, at about the time that the steady-state eye-velocity was reached. The rVOR began with a nearly head-fixed axis (0.08+/-0.04), appropriate for full retinal image stabilization, followed by a gradual increase of the tilt-angle slope to 0.31+/-0.02. Thus, differences between pursuit and the rVOR with respect to Listing's law can be seen from the onset of transient responses, although in both cases eye-position-dependent torsion increases with time. This temporal evolution of the axis of eye-velocity may involve the velocity-storage mechanism. PMID- 16307249 TI - No automatic pilot for visually guided aiming based on colour. AB - It has been claimed that visually guided limb movements are automatically corrected in response to a change in target location but not when the same change in target is cued through a colour switch (Pisella et al. 2000). These findings were based solely on limb endpoint data. Here we examine the kinematic trajectory of the hand during the entire movement. Participants pointed rapidly to a target object that could change position either by changing spatial location, or by switching colour with a second object. Participants performed in two instructional conditions: a "go" condition to index intentional movements and a "stop" condition in which failures to stop pointing indexed automatic limb guidance. Kinematic analysis indicated efficient intentional pointing in both location and colour change conditions. However, only targets that changed spatial location elicited involuntary limb modifications and these occurred within 150 ms of the change. This conclusion held even after baseline differences in the efficiency of processing colour-defined targets were taken into account, thereby strengthening the claim of a strongly automatic pilot for visually guided limb movements. PMID- 16307250 TI - Contribution of tactile feedback from the hand to the perception of force. AB - A force-matching task was used to study the influence of constraining tactile information from the hand on the perception of forces generated with the index finger flexors, the palmar prehensile grasp and with the elbow flexors. Subjects generated the same reference forces (2-10 N) with each muscle group and matched these using the corresponding muscle group in the other arm. Force perception was studied under normal conditions and when the tactile feedback from the fingertips was constrained through the use of rigid finger splints. There was no difference between the three muscle groups when matching forces under normal conditions, but when spatial tactile information from the fingertips was attenuated forces were underestimated in perceived magnitude as compared to control conditions. These results suggest that the perception of force is influenced by tactile cues that convey information about the contact surface and that distributed spatial force cues are normally used in the perception of forces generated by the hand. PMID- 16307251 TI - Entrainment to extinction of physiological tremor by spindle afferent input. AB - In this study the systematic modulation of wrist flexor muscle activity by imposed joint movement was examined. Ten subjects maintained a constant contraction level (25% of the maximum; trial duration: 20 s) in flexor carpi radialis while their wrists were perturbed with 50 different quasi-sinusoidal signals (frequency range: 0.5-9.5 Hz; amplitude: 0.3-4.2 degrees ). The frequency spectra of wrist position and the rectified and filtered electromyogram (EMG) were determined. The muscle activity was only weakly entrained to imposed movements of small amplitude and low frequency, as shown by a small peak in the EMG spectrum at the frequency of movement, while the most prominent peak in the spectrum was between 9 and 15 Hz, corresponding to the frequency range of physiological tremor. The entrainment of muscle activity increased markedly as the amplitude and frequency of the imposed movement increased, to the point of saturation of modulation and harmonic peaks in the spectrum. In parallel with this increase in entrainment, the 9-15 Hz tremor peak was progressively extinguished. The results are consistent with a coupled oscillator model in which the central oscillatory source(s) of tremor became fully entrained to the imposed movement at the highest amplitudes and frequencies. Such coupling depends on communication between the external forcing oscillator and the central oscillator(s), the I (a) afferent signal from the imposed movement being the most likely candidate to provide the entraining signal for the central oscillator(s). PMID- 16307252 TI - Multisensory control of human upright stance. AB - The interaction of different orientation senses contributing to posture control is not well understood. We therefore performed experiments in which we measured the postural responses of normal subjects and vestibular loss patients during perturbation of their stance. Subjects stood on a motion platform with their eyes closed and auditory cues masked. The perturbing stimuli consisted of either platform tilts or external torque produced by force-controlled pull of the subjects' body on a stationary platform. Furthermore, we presented trials in which these two stimuli were applied when the platform was body-sway referenced (i.e., coupled 1:1 to body position, by which ankle joint proprioceptive feedback is essentially removed). We analyzed subjects' postural responses, i.e., the excursions of their center of mass (COM) and center of pressure (COP), using a systems analysis approach. We found gain and phase of the responses to vary as a function of stimulus frequency and in relation to the absence versus presence of vestibular and proprioceptive cues. In addition, gain depended on stimulus amplitude, reflecting a non-linearity in the control. The experimental results were compared to simulation results obtained from an 'inverted pendulum' model of posture control. In the model, sensor fusion mechanisms yield internal estimates of the external stimuli, i.e., of the external torque (pull), the platform tilt and gravity. These estimates are derived from three sensor systems: ankle proprioceptors, vestibular sensors and plantar pressure sensors (somatosensory graviceptors). They are fed as global set point signals into a local control loop of the ankle joints, which is based on proprioceptive negative feedback. This local loop stabilizes the body-on-foot support, while the set point signals upgrade the loop into a body-in-space control. Amplitude non-linearity was implemented in the model in the form of central threshold mechanisms. In model simulations that combined sensor fusion and thresholds, an automatic context specific sensory re-weighting across stimulus conditions occurred. Model parameters were identified using an optimization procedure. Results suggested that in the sway-referenced condition normal subjects altered their postural strategy by strongly weighting feedback from plantar somatosensory force sensors. Taking this strategy change into account, the model's simulation results well paralleled all experimental results across all conditions tested. PMID- 16307253 TI - Spatial attention: more than intrinsic alerting? AB - It has been proposed that the right hemisphere alerting network co-activates, either directly or via the brainstem, the attention system in the parietal cortex involved in spatial attention. The observation that impaired alertness and sustained attention can predict the outcome of neglect might suggest such a relationship, too. In the present fMRI study, we intended to analyse and compare the functional anatomy of two attentional conditions both involving intrinsic (endogenous) alerting and fixation but differing with respect to the degree of spatially distributed attention by using the same paradigm under two different attentional conditions. In a group of ten participants, both a focused and a distributed visuospatial attention condition evoked similar patterns of activation in dorsolateral prefrontal regions, in the anterior cingulate gyrus, in the superior and inferior parietal cortex as well as in the superior temporal gyrus and in the thalamus. These activation foci were stronger in the right hemisphere under both conditions. After subtraction of the alertness condition with focused spatial attention, distributed spatial attention with stimuli appearing at unpredictable locations within both visual fields induced additional bilateral activations only in the left and right superior parietal cortex and in the right precuneus suggesting that these regions are specific for a more widespread dispersion of spatial attention. PMID- 16307254 TI - Heterogeneity of the developmental patterns of neurotrophin protein levels among neocortical areas of macaque monkeys. AB - Based on morphological and physiological characteristics, the mammalian neocortex is divided into various neocortical areas and its diversity is prominent in the primates including humans. These neocortical areas are constructed during development, but the details of the developmental events remain unclear, especially at the molecular level. We measured the mRNA and protein levels of neurotrophins in various neocortical areas of developing rhesus monkeys. The expression patterns of both the neurotrophin-3 (NT-3) mRNA and the protein showed area differences. In the sensory and motor areas, NT-3 mRNA and protein levels had started to decline by a week after birth. In contrast, the levels declined after the third postnatal week in the association neocortical areas. The level of brain-derived neurotrophic factor (BDNF) protein changed in an area-dependent manner during development, but that of mRNA did not. The decline of the BDNF protein level started earlier in the sensory and motor neocortical areas than in the association neocortical areas, suggesting that sensory and motor neocortical areas develop earlier than the association areas in terms of the developmental changes in neurotrophins. PMID- 16307255 TI - The control of vertical saccades in aged subjects. AB - In real life we produce vertical saccades at different distances and eccentricities, and while our fixation is more or less actively engaged. The goal of this study is to examine vertical saccades in aged and young subjects, taking into consideration all these parameters. Eleven adults (20-28 years) and 11 aged subjects (63-83 years) were recruited. We used LED targets at 7.5 degrees or 15 degrees, up or down in four conditions: gap and overlap tasks, each done at two distances-at near (40 cm) and at far (150 cm). In the gap task fixation target extinguishes prior to target onset, while in the overlap condition it stays on after target onset; consequently, visual attention and fixation are employed differently in the two tasks. Eye movements were recorded with the Chronos video eye tracker. Results showed that vertical saccades were longer for aged subjects than for young adults under almost all conditions. For both aged and young subjects, latencies were shorter under the gap than under the overlap task. Latencies for eccentric targets at 15 degrees were significantly longer than those at 7.5 degrees but for aged subjects only; this effect was more pronounced for upward saccades under the overlap condition. Express type of latencies (80 120 ms) occurred frequently in the gap task and at similar rates for young adults (16%) and aged subjects (12%); in the overlap task express latencies were scarce in young adults (0.4%) and aged subjects (1.8%). Age deteriorates the ability to trigger regular volitional saccades but not the ability to produce express type of saccades. Latency increase with aging is attributed to the degeneration of central areas, e.g. oculomotor cortical areas involved in the initiation of vertical saccades. PMID- 16307256 TI - Longer fixation duration while viewing face images. AB - The spatio-temporal properties of saccadic eye movements can be influenced by the cognitive demand and the characteristics of the observed scene. Probably due to its crucial role in social communication, it is argued that face perception may involve different cognitive processes compared with non-face object or scene perception. In this study, we investigated whether and how face and natural scene images can influence the patterns of visuomotor activity. We recorded monkeys' saccadic eye movements as they freely viewed monkey face and natural scene images. The face and natural scene images attracted similar number of fixations, but viewing of faces was accompanied by longer fixations compared with natural scenes. These longer fixations were dependent on the context of facial features. The duration of fixations directed at facial contours decreased when the face images were scrambled, and increased at the later stage of normal face viewing. The results suggest that face and natural scene images can generate different patterns of visuomotor activity. The extra fixation duration on faces may be correlated with the detailed analysis of facial features. PMID- 16307257 TI - Sensory deprivation and balance control in idiopathic scoliosis adolescent. AB - Balance control is influenced by the availability and integrity of sensory inputs as well as the ability of the balance control mechanisms to tailor the corrective action to the gravitational torque. In this study, to challenge balance control, visual and ankle proprioceptive information were perturbed (eyes closed and/or tendon vibration). We masked sensory inputs in order: (1) to test the hypothesis that adolescent idiopathic scoliosis (AIS), compared to healthy adolescent, relies more on ankle proprioception and/or visual inputs to regulate balance and (2) to determine whether it is the variation or the amplitude of the balance control commands of AIS that leads to greater body sway oscillations during sensory deprivation. By manipulating the availability of the sensory inputs and measuring the outcomes, center of pressure (CP) range and velocity variability, we could objectively determine the cost of visual and/or ankle proprioception deprivation on balance control. The CP range was larger and the root mean square (RMS) of the CP velocity was more variable for AIS than for control participants when ankle proprioception was perturbed. This was observed regardless of whether vision was available or not. The analysis of the sway density curves revealed that the amplitude rather than the variation of the balance control commands was related to a larger CP range and greater RMS CP velocity for AIS. The present results suggest that AIS, compared to control participants, relies much more on ankle proprioception to control the amplitude of the balance control commands. PMID- 16307258 TI - Aging and inhibitory errors on a motor shift of set task. AB - Inhibitory functions are key mechanisms underlying age-related cognitive decline (Park and Gutchess in Cognitive aging: a primer, Psychology Press, Hove 2000), yet how these influence the control of action has not been fully investigated. Using 134 older (age 60-88) and 133 younger adults (age 20-59), we investigated in a motor analogy of the WCST the inhibition of a primed movement plan in favour of a novel one. Although 10% of older adults performed similarly to young adults, the majority failed to inhibit by the sixties, 10-20 years earlier than documented for the WCST (Lezak in Neurological Assessment, Oxford University Press, New York 1995; Haaland et al. in J Gerontol 33:345-346 1987). Around 40% failed to learn on the second attempt, and of these, the majority in their sixties to eighties failed to learn eventually. Implications are discussed for neuropsychological theory and everyday interventions. PMID- 16307259 TI - Carbofuran-induced neurochemical and neurobehavioral alterations in rats: attenuation by N-acetylcysteine. AB - Carbofuran, a widely used carbamate pesticide, has been reported to cause neurotoxicity. However, the underlying mechanisms involved in carbofuran neurotoxicity are not well understood. The present study was envisaged to investigate the possible role of oxidative stress in carbofuran neurotoxicity and to evaluate the protective effects of N-acetylcysteine (NAC). Acetylcholinesterase activity was significantly inhibited in all the regions of brain after carbofuran exposure (1 mg/kg body weight, orally, for 28 days). NAC, on the other hand, was found to partially restore the activity of acetylcholinesterase in carbofuran treated animals. Carbofuran exposure resulted in increased lipid peroxidation (LPO) in brain regions accompanied by decreased levels of glutathione. NAC administration to the carbofuran exposed animals lowered LPO along with partial repletion in glutathione levels. Concomitantly, the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase were significantly decreased after carbofuran exposure, while no significant change in the activity of glutathione-S-transferase was observed. NAC treatment to carbofuran treated rats resulted in protective effect on the activities of these enzymes. Marked impairment in the motor function was seen following carbofuran exposure, which is evident by significant decrease in the retention time of the rats on rotating rods. Cognitive deficits were also seen after carbofuran exposure as indicated by the significant decrease in active avoidance response. NAC treatment significantly improved the carbofuran-induced neurobehavioral deficits. The results clearly demonstrate that carbofuran exerts its neurotoxic effects by accentuating oxidative stress and suggest neuroprotective role of NAC in carbofuran neurotoxicity. PMID- 16307260 TI - An object-centred reference frame for control of grasping: effects of grasping a distractor object on visuomotor control. AB - Previous evidence based on perceptual integration and arbitrary responses suggests extensive cross-modal links in attention across the various modalities. Attention typically shifts to a common location across the modalities, despite the vast differences in their initial coding of space. An issue that remains unclear is whether or not these effects of multisensory coding occur during more natural tasks, such as grasping and manipulating three-dimensional objects. Using kinematic measures, we found strong effects of the diameter of a grasped distractor object on the aperture used to grasp a target object at both coincident and non-coincident locations. These results suggest that interference effects can occur between proprioceptive and visuomotor signals in grasping. Unlike other interference effects in cross-modal attention, these effects do not depend on the spatial relation between target and distractor, but occur within an object-based frame of reference. PMID- 16307261 TI - Learning and transfer of bimanual multifrequency patterns: effector-independent and effector-specific levels of movement representation. AB - Current behavioural theories consider that during motor learning, an effector independent memory representation of the acquired skill is built up. Using a transfer paradigm, we addressed the nature of the memory representation for a 2:1 multifrequency co-ordination task, requiring, for example, the left arm to cycle twice as fast as the right. After learning this 2:1 pattern, transfer to its converse pattern (i.e., the right arm cycles twice as fast as the left) revealed powerful evidence for negative transfer. The converse task arrangement revealed similar effects. These observations suggest a reconsideration of current viewpoints on movement representations, which emphasize effector independence. Based on the present findings, we propose a new model of motor memory, consisting of an abstract, effector-independent and an effector-specific layer. The abstract code is hypothesized to represent general spatiotemporal movement features, whereas the specific representation refers to effector-related movement commands. This concept is consistent with recent neuroscientific evidence in animal and human species, and invites a reconsideration of current behavioural theories of motor learning and memory. PMID- 16307262 TI - Visuo-motor pathways in humans revealed by event-related fMRI. AB - Whether different brain networks are involved in generating unimanual responses to a simple visual stimulus presented in the ipsilateral versus contralateral hemifield remains a controversial issue. Visuo-motor routing was investigated with event-related functional magnetic resonance imaging (fMRI) using the Poffenberger reaction time task. A 2 hemifield x 2 response hand design generated the "crossed" and "uncrossed" conditions, describing the spatial relation between these factors. Both conditions, with responses executed by the left or right hand, showed a similar spatial pattern of activated areas, including striate and extrastriate areas bilaterally, SMA, and M1 contralateral to the responding hand. These results demonstrated that visual information is processed bilaterally in striate and extrastriate visual areas, even in the "uncrossed" condition. Additional analyses based on sorting data according to subjects' reaction times revealed differential crossed versus uncrossed activity only for the slowest trials, with response strength in infero-temporal cortices significantly correlating with crossed-uncrossed differences (CUD) in reaction times. Collectively, the data favor a parallel, distributed model of brain activation. The presence of interhemispheric interactions and its consequent bilateral activity is not determined by the crossed anatomic projections of the primary visual and motor pathways. Distinct visuo-motor networks need not be engaged to mediate behavioral responses for the crossed visual field/response hand condition. While anatomical connectivity heavily influences the spatial pattern of activated visuo-motor pathways, behavioral and functional parameters appear to also affect the strength and dynamics of responses within these pathways. PMID- 16307263 TI - At what stage of neural processing do perspective depth cues make a difference? AB - The present study investigated the cortical processing of three-dimensional (3D) perspective cues in humans, to determine how the brain computes depth from a bidimensional retinal image. We recorded visual evoked potentials in 12 subjects in response to flat and in-perspective stimuli, which evoked biphasic potentials over posterior electrodes. The first, positive component (P1, at 90 ms) was not sensitive to perspective, while the second, negative peak (N1 at approximately 150 ms) was significantly larger for 3D stimuli, regardless of attention. The amplitude increase due to perspective was seen on all posterior electrodes, but was largest over the right hemisphere, particularly at parietal sites. Source modeling low-resolution electromagnetic tomography (LORETA) confirmed that among the different areas participating in two- and three-dimensional stimuli processing, the right parietal source is the most enhanced by perspective depth cues. We conclude that the extraction of depth from perspective cues occurs at a second level of stimulus processing, by increasing the activity of the regions involved in 2D stimuli processing, particularly in the right hemisphere, possibly through feedback loops from higher cortical areas. These modulations would participate in the fine-tuned analysis of the 3D features of stimuli. PMID- 16307264 TI - The effect of force and conformance on tactile intensive and spatial sensitivity. AB - The effect of force on intensive and spatial processing was examined with three measures of tactile sensitivity. One of the measures based on intensive cues is the smooth-grooved (SM/GV) task, and the two other measures based on spatial cues are the grating orientation and gap detection tasks. Measures were made at two locations that vary in sensitivity and in the density of innervation of the primary afferent fibers, the right index fingerpad and the palmar surface of the proximal phalanx (fingerbase). At each location, psychometric functions were generated for each of the three measures for two forces (50 and 200 g). The results indicated that increasing force led to marked improvement on the task that relied on intensive cues; however, on the tasks that relied on spatial cues, force had no effect on performance. Biomechanical measures were made of the depth to which the skin invades the grooves of the contactors (conformance) at the two test sites, with the two forces, and with different groove widths. Conformance was found to be a joint function of force and groove width. Further, performance on the SM/GV task could be predicted by the amount of conformance. The psychophysical results are consistent with the view that increasing conformance increases neural activity in the primary afferent fibers, and that this increase in neural activity improves SM/GV performance, but has little effect on the quality of the spatial image. PMID- 16307265 TI - Centrifugal regulation of task-relevant somatosensory signals to trigger a voluntary movement. AB - Many previous papers have reported the modulation of somatosensory evoked potentials (SEPs) during voluntary movement, but the locus and mechanism underlying the movement-induced centrifugal modulation of the SEPs elicited by a task-relevant somatosensory stimulus still remain unclear. We investigated the centrifugal modulation of the SEPs elicited by a task-relevant somatosensory stimulus which triggers a voluntary movement in a forewarned reaction time task. A pair of warning (S1: auditory) and imperative stimuli (S2: somatosensory) was presented with a 1 s interstimulus interval. Subjects were instructed to respond by moving the hand ipsilateral or contralateral to the somatosensory stimulation which elicits the SEPs. In four experiments, the locus and selectivity of the SEPs' modulation, the contribution of cutaneous afferents and the effect of contraction magnitude were examined, respectively. A control condition where subjects had no task to perform was compared to several task conditions. The amplitude of the frontal N30, parietal P30, and central P25 was decreased and that of the long latency P80 and N140 was increased when the somatosensory stimuli triggered a voluntary movement of the stimulated finger compared to the control condition. The N60 decreased with the movement of any finger. These results were considered to be caused by the centrifugal influence of neuronal activity which occurs before a somatosensory imperative stimulus. The present findings did not support the hypothesis that the inhibition of afferent inputs by descending motor commands can occur at subcortical levels. A higher contraction magnitude produced a further attenuation of the amplitude of the frontal N30, while it decreased the enhancement of the P80. Moreover, the modulation of neuronal responses seems to result mainly from the modulation of cutaneous afferents, especially from the moved body parts. In conclusion, the short- and long-latency somatosensory neuronal activities evoked by task-relevant ascending afferents from the moved body parts are regulated differently by motor-related neuronal activities before those afferent inputs. The latter activities may be associated with sensory gain regulation related to directing attention to body parts involved in the action. PMID- 16307266 TI - Alteration of thyroid hormone homeostasis by antiepileptic drugs in humans: involvement of glucuronosyltransferase induction. AB - RATIONALE: The aim of this review article is to analyse which antiepileptic drugs (AEDs) alter thyroid hormone homeostasis in humans and when this can be explained, at least partially, by the induction of the glucuronoconjugation pathways. METHODS: Electronic databases were searched which have provided more than 300 articles. These have been integrated with fundamental books and personal information by experts in the different areas examined. RESULTS: Alteration of thyroid hormone homeostasis by phenobarbital/primidone, phenytoin, and carbamazepine clearly occurs in humans. However, it is not associated with thyroid-stimulating hormone (TSH) increase and the clinical significance of altered serum concentrations of thyroid hormones by these antiepileptic drugs has remained unclear. The published information on the effect of the other antiepileptic drugs examined in this review article on thyroid hormones is lacking (felbamate, pregabalin, zonisamide) or limited. Oxcarbazepine appears to have some effects. Topiramate would need further investigations as well as gabapentin. Levetiracetam, tiagabine, vigabatrine, and lamotrigine do not alter at all, or only minimally, thyroid hormone homeostasis. CONCLUSION: Concerning the antiepileptic drugs which alter thyroid hormone homeostasis, it is highly probable that the mechanism of induction of uridine diphosphate glucuronosyltransferases (UGT) is involved, at least partially, in such an alteration. However, it is not possible to estimate the relative contribution of the UGT induction by these drugs on the total alteration observed in thyroid hormone levels, as other mechanisms not investigated, or not examined in the present article, could contribute. PMID- 16307267 TI - Patterns of drug prescription in a geriatric evaluation and management unit as compared with the general medical wards: a randomised study. AB - OBJECTIVE: This study's objective was to determine whether patients treated in a geriatric evaluation and management unit (GEMU) had a more appropriate drug profile than patients treated in the general medical wards (MW). METHODS: Frail elderly patients admitted as emergencies to the medical department were randomised to treatment in the GEMU (n=127) or MW (n=127). Drugs used at inclusion and discharge were registered retrospectively and analysed with regard to polypharmacy, number of drugs withdrawn or started, potential drug-drug interactions (DDIs), number of anticholinergic drugs prescribed, and the number of inappropriate drug prescriptions according to Beers' criteria. Utilisation of psychotropic and cardiovascular drugs was compared in detail according to prespecified hypotheses. RESULTS: The number of patients with polypharmacy did not differ significantly between the GEMU and MW. The median number of scheduled drugs withdrawn per patient was higher in the GEMU than in the MW (p=0.005). Drugs with anticholinergic effects (p=0.003); cardiovascular drugs (p<0.001), particularly digitalis glycosides (p<0.001); and antipsychotic drugs (p=0.009) were withdrawn more often in the GEMU. The median number of scheduled drugs started was higher in the GEMU than in the MW (p=0.03). In particular, antidepressants (p<0.001) and estriol (p=0.001) were started more often in the GEMU than in the MW. Fewer GEMU than MW patients had potential DDIs at discharge (p=0.009). CONCLUSION: Drug treatment in the GEMU as compared with the MW was more appropriate in terms of prescription of fewer drugs with anticholinergic effects and fewer potential DDIs. There were distinct differences in treatment patterns of cardiovascular and psychotropic drugs. PMID- 16307268 TI - Inappropriate medication prescribing in community-dwelling elderly people living in Iran. AB - OBJECTIVES: To determine the prevalence of inappropriate medication prescribing in Middle Eastern community-dwelling elderly persons, and to determine factors associated with inappropriate prescribing. METHODS: Prescriptions from 3,000 elderly-patient visits (> or =65 years) to physicians in Isfahan, Iran were collected between September and December 2002; only the first patient visit was considered in the study. Inappropriate prescriptions were defined according to Beers' explicit criteria (1997). The presence of at least one drug-drug interaction (DDI) and at least one drug-class duplication within each patient visit was also determined. Multivariate analyses were performed to determine factors associated with receiving at least one inappropriate medication. These factors included age, gender, number of prescribed medications per visit, DDIs, duplications, and physician's number of years of experience. Similar analyses were performed to determine whether these factors were associated with a composite index (defined as having at least one of the following: receiving at least one inappropriate prescription, DDI, or duplication). RESULTS: The mean age of our cohort was 72.6 (+/-5.7) years, and 1,735 (57.8%) were females. A total of 829 (27.6%) patients received at least one inappropriate prescription, 285 (9.5%) had at least one DDI during the visit, and 746 (24.9%) had at least one duplication during the visit. The three most inappropriately prescribed medication classes were antihistamines (29%), non-steroidal anti-inflammatory agents (23%) and benzodiazepines (16%). In multivariate analyses, the number of prescribed medications per visit was the only factor significantly associated with receiving at least one inappropriate prescription (OR 1.24; 95%CI: 1.16 1.32); analyses using the composite index as outcome gave similar results (OR: 1.70; 95%CI: 1.60-1.80). CONCLUSION: Our findings indicate a high prevalence of inappropriate prescribing in elderly patients. Given the potential severity of this problem, further steps need to be implemented to prevent this occurrence. PMID- 16307269 TI - Rapid genotyping for relevant CYP1A2 alleles by pyrosequencing. AB - OBJECTIVE: To develop a rapid and reliable screening method for identifying the relevant cytochrome P450 (CYP) 1A2 alleles CYP1A2*1D (-2467Tdel), *1F (-163A>C), and *1K (-739T>G, -729C>T, -163A>C) that are in linkage disequilibrium with the functionally relevant CYP1A2 polymorphisms and therefore are considered to be predictive for the CYP1A2 phenotype. METHODS: CYP1A2 single nucleotide polymorphisms (SNPs) -2467Tdel, -739T>G, -729C>T, and -163A>C were screened for in 495 healthy Caucasian volunteers using newly developed pyrosequencing duplex and simplex assays. Conventional sequencing of randomly selected samples served as quality control. RESULTS: Frequencies were 7.9% for CYP1A2*1D, 31.8% for *1F, and 0.4% for *1K. The observed distribution of homozygous and heterozygous carriers of the alleles corresponded to the predicted one according to the Hardy Weinberg law. It also corresponded to reported allelic frequencies from Caucasians but differed significantly from the distribution seen in other ethnicities. The most frequent haplotype was -2467T/-739T/-729C/-163A (allelic frequency 61.6%), followed by -2467T/-739T/-729C/-163C (30.5%), -2467Tdel/-739T/ 729C/-163A (5.1%), -2467Tdel/-739G/-729C/-163A (1.2%), and -2467Tdel/-739T/-729C/ 163C (1.1%). Complete linkage disequilibrium (value of D' nearly 1) existed between -2467Tdel, -739T>G, and -729C>T and between -729T>G and -163A>C. CONCLUSIONS: Pyrosequencing facilitates rapid and reliable detection of those CYP1A2 alleles that, based on current knowledge, can be considered predictive for the CYP1A2 phenotype. PMID- 16307270 TI - Evaluation of rat and rabbit sera lipoproteins in experimentally induced hyperlipidemia by analytical ultracentrifugation. AB - Animals of various species are widely used as models with which to study atherosclerosis and the lipoprotein metabolism. The objective of this study was to investigate the lipoprotein profiles in Wistar rats and New Zealand white rabbits with experimentally induced hyperlipidemia by means of ultracentrifugation. The Schlieren curves were utilized to compare suckling and adult rat sera to determine whether aging causes alterations in lipoprotein profiles. A striking feature of the data is the high concentration of low-density lipoproteins (LDL), (>5.2 mmol/l cholesterol) in the 2-week old rat serum pool which was greatly decreased in the 3-weeks rat serum pool (<1.3 mmol/l cholesterol). Additional experiments were performed to permit a direct comparison of the amounts of lipoprotein present in rat sera in experimental hyperlipidemia post-Triton WR 1339 administration. Rapid changes in concentrations in very low density lipoproteins (VLDL), LDL and high-density lipoproteins (HDL) were observed after Triton injection. The administration of Triton WR 1339 to fasted rats resulted in an elevation of serum cholesterol levels. Triton physically alters VLDL, rendering them refractive to the action of lipolytic enzymes in the blood and tissues, preventing or delaying their removal from the blood. Whereas the VLDL concentration was increased markedly, those of LDL and HDL were decreased at 20 h after Triton treatment. Rabbits were fed a diet containing 2% cholesterol for 60 days to develop hyperlipidemia and atheromatous aortic plaques. A combination of preparative and analytical ultracentrifugation was used to investigate of LDL aliquots, to prepare radioactive-labeled lipoproteins and to study induced hyperlipidemia in rabbits. Analytical ultracentrifugation was applied to investigate the LDL flotation peaks before and after cholesterol feeding of rabbits. Modified forms of LDL were detected in the plasma of rabbits with experimentally induced atherosclerosis. ApoB-containing particles, migrating as LDL, intermediate density lipoproteins and VLDL were the most abundant lipoproteins. Gamma camera in vivo scintigraphy on rabbits with radiolabeled lipoproteins revealed visible signals corresponding to atherosclerotic plaques of the aorta and carotid arteries. PMID- 16307271 TI - Microbial hydroxylation of imidacloprid for the synthesis of highly insecticidal olefin imidacloprid. AB - Microorganisms that bring about the aerobic transformation of imidacloprid (IMI) were isolated and screened, and the microbial regio- and stereoselective hydroxylation of IMI was studied. Some bacteria and fungi transformed IMI to 5 hydroxyl IMI. Bacterium Stenotrophomonas maltophilia CGMCC 1.1788 resting cells transformed IMI into R-5-hydroxyl IMI at the highest conversion rate. The enzyme catalyzed the stereoselective hydroxylation at position C12 of IMI in the imidazolidine ring. Under acidic conditions, 5-hydroxyl IMI was converted into olefin IMI in high molar conversion yield. The olefin IMI exhibited about 19 and 2.2 times more insecticidal efficacy than IMI against horsebean aphid imago and nymph, respectively, and about 1.4 times more active than IMI against brown planthopper imago. The transformation rate of IMI by resting cells of S. maltophilia CGMCC 1.1788 was promoted significantly by some carbohydrates and organic acids. The reaction medium with 5% sucrose resulted in 8.3 times greater biotransformation yield as compared with that without sucrose. PMID- 16307272 TI - Prion variant maintained only at high levels of the Hsp104 disaggregase. AB - The yeast prion [PSI(+)] is a self-perpetuating aggregated isoform of the translation termination factor Sup35. [PSI ( + )] propagation is promoted by moderate levels and antagonized by high levels of the chaperone Hsp104. In agreement with the model postulating that excess Hsp104 acts on [PSI ( + )] by disaggregating prion polymers, we show that an increase in Sup35 levels, accompanied by an increase in size of prion aggregates, also partially protects [PSI(+)] from elimination by excess Hsp104. Despite retention of [PSI(+)], excess Hsp104 decreases toxicity of overproduced Sup35 in [PSI(+)] strains. A heritable variant of [PSI(+)], which has been isolated and is maintained only in the presence of increased levels of Hsp104, is characterized by an abnormally large aggregate size, and exhibits an altered response to overproduction of the Hsp70 chaperone Ssa1. These features resemble the previously described prion generated by a deletion derivative of Sup35, but are not associated with any sequence alteration and are controlled exclusively at the protein level. Our data provide a proof of the existence of conditionally stable prion variants maintained only at altered levels of Hsps, that could in principle be beneficial if the normal cellular function of a prion protein becomes detrimental to the cell in such conditions. PMID- 16307273 TI - Expression of ICAM-1, ICAM-2, NCAM-1 and VCAM-1 by human synovial cells exposed to Borrelia burgdorferi in vitro. AB - The interaction of resident tissue cells with migratory inflammatory cells is essential for the recruitment of immune effector cells to inflammatory sites. The sustained expression of adhesion molecules in the synovium of patients with chronic Lyme arthritis seems to contribute to this chronic inflammation. Whether cell adhesion molecules influence the early steps of Borreliosis is unclear. Therefore, we examined the expression of ICAM-1, ICAM-2, VCAM-1 and NCAM-1 in synovial cells exposed to two different Borrelia burgdorferi sensu stricto strains Geho and B31. The mRNA expression of ICAM-1, ICAM-2, VCAM-1 and NCAM-1 was not changed in synovial cells exposed to B31. Whereas ICAM-2 and VCAM-1 was upregulated, NCAM-1 mRNA was downregulated and ICAM-1 mRNA was unchanged by strain Geho. The ICAM-1 protein expression on the synovial cell surface was downregulated by both strains. Differential regulation of adhesion molecule mRNA, and subsequent high turnover or elevated shedding from the cell membrane may contribute to early pathogenesis in Lyme arthritis. PMID- 16307274 TI - Familial Mediterranean fever and ankylosing spondylitis in a patient with juvenile idiopathic arthritis: A case report and review of the literature. AB - The association of familial Mediterranean fever (FMF) with juvenile idiopathic arthritis (JIA) or ankylosing spondylitis (AS), most commonly with negative HLA B27 antigen, was described in several previous reports, although the pathogenic mechanism of this association still remains unknown. Herein we report an uncommon association of FMF with HLA-B27 positive AS as an occasional coincidence in a patient who had been diagnosed as having JIA 23 years previously. PMID- 16307275 TI - Day-night specific binding of 2-[125I]iodomelatonin and melatonin content in gill, small intestine and kidney of three fish species. AB - Some of melatonin's (Mel) well-established physiological effects are mediated via high-affinity cell-membrane receptors belonging to the superfamily of G-protein coupled receptors. Specific binding of ligand 2-[(125)I]iodomelatonin, using membrane preparations from osmoregulatory tissues of flounder, rainbow trout and sea bream, together with Mel concentrations in the tissues and plasma were studied. The kidney, gill and small intestine samples were collected during the day and at night. The dissociation constants (K (d)) and maximal binding densities (B (max)) were calculated for each tissue at 11:00 and 23:00 h. The binding sites with K (d) values in the tissues in the picomolar range indicated the high affinity. K (d) and B (max) values were tissue- and species-dependent. The GTP analogue [Guanosine 5'-O-(3-thiotriphosphate)] treatment significantly reduced the B (max) value, indicating that the 2-[(125)I]iodomelatonin-binding sites are probably coupled to a G-protein. No daily variations in K (d) and B (max) values were observed. These are the first studies of the presence of 2 [(125)I]iodomelatonin-binding sites in the small intestine, kidney tubule and gill of fish. The data strongly suggest new potential targets for Mel action and the influence of Mel on water/ion balance in fish. The intestine seems to be a site of Mel synthesis and/or an active accumulation of the hormone. PMID- 16307276 TI - Rectovestibular fistula with vaginal malformations. AB - Anorectal malformations are associated with other anomalies among which vaginal malformations are occasionally encountered and may go unnoticed by the primary physician. Between January 1998 and December 2003, 563 cases of anorectal malformations were managed in Pediatric Surgery Department, Chittagong Medical College & Hospital, Chittagong, Bangladesh. Among these, five cases of rectovestibular fistula were associated with vaginal malformations. In this retrospective study age, physical findings, operative findings and procedures, outcome of operation and postoperative follow-up were evaluated. Mean age at presentation was 8.67 years (range 2 months to 17 years). All cases referred as rectovaginal fistula and vaginal anomalies (atresia to agenesis) were detected after thorough examination. Initial pelvic colostomy was done in all patients. Cases 1, 3 and 4 had distal vaginal agenesis and underwent posterior sagittal anorectovaginoplasty. In case 5, atretic vaginal duplication was found with didelphic hypoplastic uterus and absent left kidney. Case 2 (vaginal atresia) operated elsewhere is waiting for definitive surgery. Colostomy closure was done in four cases. The third patient had already married and conceived. Bowel habits are regular in all except the second patient. In females, a thorough understanding of anorectal malformations is necessary to identify the association with vaginal anomalies and awareness of this association will lead to earlier diagnosis and appropriate operative measures. PMID- 16307277 TI - Influence of the site of small bowel resection on intestinal epithelial cell apoptosis. AB - Massive small bowel resection (SBR) results in a significant increase in intestinal epithelial cell (EC) proliferation as well as apoptosis. Because the site of SBR (proximal (P) vs. distal (D)) affects the degree of intestinal adaptation, we hypothesized that different rates of EC apoptosis would also be found between P-SBR and D-SBR models. Wild-type C57BL/6J mice underwent: (1) 60% P-SBR, (2) 60% D-SBR, or (3) SHAM-operation (transaction-reanastomosis) at the mid-gut point. Mice were sacrificed after 7 days. EC apoptosis was measured by TUNEL staining. EC-related apoptotic gene expression including intrinsic and extrinsic pathways was measured with reverse transcriptase-polymerase chain reaction. Bcl-2 and bax protein expression were analyzed by Western immunoblotting. Both models of SBR led to significant increases in villus height and crypt depth; however, the morphologic adaptation was significantly higher after P-SBR compared to D-SBR (P<0.01). Both models of SBR led to significant increases in enterocyte apoptotic rates compared to respective sham levels; however, apoptotic rates were 2.5-fold higher in ileal compared to jejunal segments (P<0.01). P-SBR led to significant increases in bax (pro-apoptotic) and Fas expression, whereas D-SBR resulted in a significant increase in TNF-alpha expression (P<0.01). EC apoptosis seems to be an important component of intestinal adaptation. The significant difference in EC apoptotic rates between proximal and distal intestinal segments appeared to be due to utilization of different mechanisms of action. PMID- 16307278 TI - In situ assessment of oxidant and nitrogenic stress in bleomycin pulmonary fibrosis. AB - Reactive oxygen species (ROS) and nitric oxide (NO) have a role in the development of pulmonary fibrosis after bleomycin administration. The ROS production induces an antioxidant response, involving superoxide dismutases (SODs), catalase, and glutathione peroxidases. We compared in situ oxidative burden and antioxidant enzyme activity in bleomycin-injured rat lungs and normal controls. ROS expression and catalase, glucose-6-phosphate-dehydrogenase (G6PHD), and NOS/NADPH-diaphorase activity were investigated by using histochemical reactions. Nitric oxide synthase (e-NOS and i-NOS) and SOD (MnSOD, Cu/ZnSOD, ECSOD) expression was investigated immunohistochemically. After treatment ROS production was enhanced in both phagocytes and in type II alveolar epithelial cells. Mn, Cu/Zn, and ECSOD were overexpressed in parenchymal cells, whereas interstitium expressed ECSOD. Catalase and G6PHD activity was moderately increased in parenchymal and inflammatory cells. NOS/NADPH-d activity and i-NOS expression increased in alveolar and bronchiolar epithelia and in inflammatory cells. It can be suggested that the concomitant activation of antioxidant enzymes is not adequate to scavenge the oxidant burden induced by bleomycin lung damage. Inflammatory cells and also epithelial cells are responsible of ROS and NO production. This oxidative and nitrosative stress may be a substantial trigger in TGF-beta1 overexpression by activated type II pneumocytes, leading to fibrotic lesions. PMID- 16307279 TI - Living high-training low: tolerance and acclimatization in elite endurance athletes. AB - The "living high-training low" (LHTL) model is frequently used to enhance aerobic performance. However, the clinical tolerance and acclimatization process to this intermittent exposure needs to be examined. Forty one athletes from three federations (cross-country skiers, n=11; swimmers, n=18; runners, n=12) separately performed a 13 to 18-day training at the altitude of 1,200 m, by sleeping either at 1,200 m (CON) or in hypoxic rooms (HYP), with an O2 fraction corresponding to 2,500 m (5 nights for swimmers and 6 for skiers and runners), 3,000 m (6 nights for skiers, 8 for swimmers and 12 for runners) and 3,500 m (6 nights for skiers). Measurements performed before, 1 or 15 days after training were ventilatory response (HVRe) and desaturation (deltaSaO2e) during hypoxic exercise, an evaluation of cardiac function by echocardiography, and leukocyte count. Lake Louise AMS score and arterial O2 saturation during sleep were measured daily for HYP. Subjects did not develop symptoms of AMS. Mean nocturnal SaO2 decreased with altitude down to 90% at 3,500 m and increased with acclimatization (except at 3,500 m). Leukocyte count was not affected except at 3,500 m. The heart function was not affected by LHTL. Signs of ventilatory acclimatization were present immediately after training (increased HVRe and decreased deltaSaO2e) and had disappeared 15 days later. In conclusion, LHTL was well tolerated and compatible with aerobic training. Comparison of the three patterns of training suggests that a LHTL session should not exceed 3,000 m, for at least 18 days, with a minimum of 12 h day(-1) of exposure. PMID- 16307280 TI - Bone status in elite male runners. AB - The aim of our study was to compare long distance runners to body mass index (BMI)- and age-matched healthy controls with respect to bone parameters at all relevant loaded and nonloaded skeletal sites. Furthermore, we assessed the effect of running volume on bone parameters. Twenty elite male runners (21.1 km<1:15 h; volume >75 km/week/year) participated in the study (RG), 11 age- and BMI-matched male subjects (28+/-5 years) served as nontraining controls (CG). Subjects with any medication or illness affecting bone metabolism or with a family history of osteoporosis were not included. Bone parameters at various sites (total body, lumbar spine, femoral neck/hip, calcaneus) were measured by dual energy X-ray (DXA), quantitative computed tomography and quantitative ultrasound. Body composition was assessed via DXA and bioimpedance analysis; nutritional parameters were determined by 5-day dietary protocols. Training variables were assessed by questionnaires. Compared with nontraining controls runners had significantly higher BMD at all loaded sites (calcaneus, lower limbs, femoral neck, pelvis, and trabecular lumbar spine). BMD at nonloaded sites (ribs, upper limbs, and skull) was slightly but not significantly higher in the runners. We observed a low (r=0.30), nonsignificant association between training volume (km/week/year) and trabecular BMD of the femoral neck, which disappeared after adjusting for age, BMI, and body fat in this group of highly trained male runners. The effect of long distance running per se on bone parameters is not deleterious. PMID- 16307281 TI - The heart rate increase at the onset of high-work intensity exercise is accelerated by central blood volume loading. AB - Using a water immersion (WI) method, the combined effect of central blood volume (CBV) loading and work intensity on the time course of heart rate (HR) at the onset of upright dynamic exercise was investigated. Seven males cranked a cycle ergometer for 12 min using their un-immersed arms at low-, moderate- and high work intensities, followed by a 12-min rest. For WI, the pre-exercise resting cardiac output increased by 36%, while HR decreased by 22% [from 76.8 (10.4) to 59.6 (9.8) beats/min]. WI also increased the high-frequency (HF, 0.15-0.40 Hz) component of the HR variability, suggesting an increased vagal activity. During the initial 2 min of the exercise period at low-work intensity, HR increased by 34.9 and 25.8% in the WI and control conditions, respectively. These were 117 and 73% at high-work intensity, indicating more accelerated HR with WI than the control. The plasma norepinephrine concentration increased less during high-work intensity exercise during WI, as compared to exercise during control conditions. In conclusion, the HR increase at the onset of high-work intensity exercise is accelerated by CBV loading but not at low intensity, possibly reflecting vago sympathetic interaction and reduced baroreflex sensitivity. PMID- 16307282 TI - Development- and tissue-specific expression of the RpoT gene family of Arabidopsis encoding mitochondrial and plastid RNA polymerases. AB - Arabidopsis thaliana possesses three RpoT genes which encode three different phage-type RNA polymerases with yet unknown function in organelle transcription: RpoTm and RpoTp, imported into mitochondria and plastids, respectively, and RpoTmp, co-targeted into both organelles. Expression of the RpoT genes was analyzed by quantitative RT-PCR, histochemical beta-glucuronidase (GUS) assays and in situ hybridization. Transcripts of all three RpoT genes accumulated to very low amounts in all organs. Surprisingly, RT-PCR revealed their highest levels in flower tissues. RpoTm transcripts were the most abundant in all organs, except mature leaves, in which RpoTp transcripts showed the highest accumulation. In the developing seedling, RpoTm::GUS and RpoTmp::GUS expression precedes that of RpoTp::GUS, the latter showing up only 7 days after germination. The RpoTm and RpoTmp promoters expressed GUS mainly in meristematic and mitochondria-rich cells such as the distal part of the root and companion cells flanking the phloem, whereas RpoTp::GUS activity was found in green tissues as the parenchyme cells of young leaves, the primary cortex of the stem, and sepals of buds and young flowers. Sites of GUS expression coincided spatially with those of in situ hybridization. Our data demonstrate an overlapping expression pattern of RpoTm and RpoTmp, and a completely differing pattern of RpoTp expression. The results suggest that RpoTm and RpoTmp recognize different types of mitochondrial promoters. The plastid polymerase RpoTp might play a major role in green tissue, i.e. in chloroplast transcription, whilst the dual-targeted RpoTmp in plastids should function mainly in the transcription of genes in non-green types. PMID- 16307283 TI - Reconstitution of cyanogenesis in barley (Hordeum vulgare L.) and its implications for resistance against the barley powdery mildew fungus. AB - Barley (Hordeum vulgare L.) produces a leucine-derived cyanogenic beta-D glucoside, epiheterodendrin that accumulates specifically in leaf epidermis. Barley leaves are not cyanogenic, i.e. they do not possess the ability to release hydrogen cyanide, because they lack a cyanide releasing beta-D-glucosidase. Cyanogenesis was reconstituted in barley leaf epidermal cells through single cell expression of a cDNA encoding dhurrinase-2, a cyanogenic beta-D-glucosidase from sorghum. This resulted in a 35-60% reduction in colonization rate by an obligate parasite Blumeria graminis f. sp. hordei, the causal agent of barley powdery mildew. A database search for barley homologues of dhurrinase-2 identified a (1,4)-beta-D-glucan exohydrolase isozyme betaII that is located in the starchy endosperm of barley grain. The purified barley (1,4)-beta-D-glucan exohydrolase isozyme betaII was found to hydrolyze the cyanogenic beta-D-glucosides, epiheterodendrin and dhurrin. Molecular modelling of its active site based on the crystal structure of linamarase from white clover, demonstrated that the disposition of the catalytic active amino acid residues was structurally conserved. Epiheterodendrin stimulated appressoria and appressorial hook formation of B. graminis in vitro, suggesting that loss of cyanogenesis in barley leaves has enabled the fungus to utilize the presence of epiheterodendrin to facilitate host recognition and to establish infection. PMID- 16307284 TI - Triacylglycerol phase and 'intermediate' seed storage physiology: a study of Cuphea carthagenensis. AB - Seeds with 'intermediate' storage physiology store poorly under cold and dry conditions. We tested whether the poor shelf life can be attributed to triacylglycerol phase changes using Cuphea carthagenensis (Jacq.) seeds. Viability remained high when seeds were stored at 25 degrees C, but was lost quickly when seeds were stored at 5 degrees C. Deterioration was fastest in seeds with high (>or=0.10 g g(-1)) and low (0.01 g g(-1)) water contents (g H(2)O g dry mass(-1)), and slowest in seeds containing 0.04 g g(-1). A 45 degrees C treatment before imbibition restored germination of dry seeds by melting crystallized triacylglycerols. Here, we show that the rate of deterioration in C. carthagenensis seeds stored at 5 degrees C correlated with the rate that triacylglycerols crystallized within the seeds. Lipid crystallization, measured using differential scanning calorimetry, occurred at 6 degrees C for this species and was fastest for seeds stored at 5 degrees C that had high and very low water contents, and slowest for seeds containing 0.04 g g(-1). Germination decreased to 50% (P50) when between 16 and 38% of the triacylglycerols crystallized; complete crystallization took from 10 to over 200 days depending on water content. Our results demonstrate interactions between water and triacylglycerols in seeds: (1) water content affects the propensity of triacylglycerols to crystallize and (2) hydration of seed containing crystallized triacylglycerols is lethal. We suggest that these interactions form the basis of the syndrome of damage experienced when seeds with intermediate storage physiologies are placed in long-term storage. PMID- 16307285 TI - Primary metabolic pathways and signal transduction in sunflower (Helianthus annuus L.): comparison of transcriptional profiling in leaves and immature embryos using cDNA microarrays. AB - The early stage of embryo development is a critical step in plant production. To identify genes with potential roles in the early sunflower seed development, a cDNA microarray approach was employed. We developed a thematic cDNA microarray containing clones representing high sequence similarities with known or predicted Arabidopsis genes implicated in different metabolic and signal transduction pathways. This 800-element cDNA array was used to compare the expression patterns in leaves and immature embryos (2 mm and 6 mm). Statistical analysis, using two step ANOVA, revealed that 143 cDNA clones can be considered as differentially expressed. Of these, 62 clones were found to be up-regulated in leaves, 81 in embryos whereas only seven clones displayed increased level of mRNA in the 6 mm embryos when compared with 2 mm embryos. The differentially expressed clones are distributed among many metabolic and signal transduction pathways. For example, genes related to fatty acid metabolism and amino acid biosynthesis exhibited preferential expression patterns in immature embryos. Also, clones potentially encoding enzymes involved in the metabolism of ascorbate and aldarate, pyruvate, propanoate and inositol, and citrate cycle were found to be up-regulated in embryos. In contrast, cDNA clones putatively involved in energy metabolism were more abundant in leaves than embryos. Clones encoding potential signal transduction components including receptors, protein kinases, protein phosphatases, and transcription factors were also identified, with preferential expression profiles in immature embryos. The expression patterns derived from this study provide initial characterization of metabolic pathways and signalling transduction networks occurring in the early stage of sunflower seed development. PMID- 16307286 TI - Increased pathogen resistance and yield in transgenic plants expressing combinations of the modified antimicrobial peptides based on indolicidin and magainin. AB - Reverse peptide of indolicidin (Rev4), a 13-residue peptide based on the sequence of indolicidin, has been shown to possess both strong antimicrobial and protease inhibitory activities in vitro. To evaluate its efficacy in vivo, we produced and evaluated transgenic tobacco (Nicotiana tabacum L.) and Arabidopsis thaliana [(L.) Heynh.] plants expressing Rev4 with different signal peptide sequences for pathogen resistance. All transgenic plants showed normal growth and development, an indication of no or low cytotoxicity of the peptide. Furthermore, the transgenic plants exhibited elevated resistance to three bacterial and two oomycete pathogens. Interestingly, tobacco plants expressing Rev4 displayed enhanced yield compared to the control as indicated by an increased biomass production by as much as 34% in two field trials. When Rev4 was coexpressed with another antimicrobial peptide, Myp30, the disease resistance levels in the transgenic Arabidopsis were enhanced. These findings suggest the potential of using these peptides to protect plants from microbial pathogens and to enhance yield. PMID- 16307287 TI - Photomorphogenetic characteristics are severely affected in nucleoside diphosphate kinase-1 (ndk-1)-disrupted mutants in Neurospora crassa. AB - We previously demonstrated that the NDK-1 (Nucleoside Diphosphate Kinase-1) point mutant, ndk-1(P72H), displays a defective phenotype in light-induced perithecial polarity in Neurospora crassa. To investigate the biological function of NDK-1 in detail, we isolated two ndk-1 mutants, ndk-1(RIP-1) and ndk-1(RIP-2), using the RIPing (repeat induced point mutation) method. Notably, we detected no accumulation of ndk-1(RIP-1) mRNA and truncated NDK-1(RIP-2) protein. The ndk 1(RIP) mutants exhibited altered morphogenesis; (1) aerial hypha was not formed with no conidium formation, (2) the mutants exhibited colonial, and very slow mycelial growth on a solid medium and by shaking culture in a liquid medium, (3) light-induced carotenoid accumulation in mutant mycelia is reduced to less than half that by wild type, (4) the mutants exhibited spiral growth of mycelia, and (5) female sterility with defective protoperithecium formation. The morphogenetic processes of 1, 3, and 5 are light induced in the wild type. Moreover, despite only 10-20% of total nucleoside diphosphate kinase activity, the accumulation of relevant transcripts in the ndk-1(RIP) mutants, such as al-1 and al-2, was similar to that of wild type. PMID- 16307288 TI - Anatomical evidence for glutamatergic transmission in primary sensory neurons and onto postganglionic neurons controlling penile erection in rats: an ultrastructural study with neuronal tracing and immunocytochemistry. AB - In male rats, the dorsal penile nerve (DPN) conveys sensory information from the genitals to the lumbosacral spinal segments of the spinal cord. DPN is the afferent limb of a reflex loop that supports reflexive erections, and that includes a network of spinal interneurons and autonomic and somatic motoneurons to the penis and perineal striated muscles. Autonomic efferent pathways to the penis relay in the major pelvic ganglion (MPG). Glutamate (Glu) is a likely candidate as a neurotransmitter of reflexive erections. Both AMPA and NMDA glutamatergic receptor subunits are present in the lumbosacral spinal cord, and AMPA and NMDA receptor antagonists block reflexive erections. In the present study, we used tract-tracing experiments combined with immunohistochemical and immunocytochemical techniques to ascertain the presence of Glu at two different levels of the network controlling reflexive erections. DPN afferents were localized in the dorsal horn of the lumbosacral cord and displayed the characteristics of either C-fibers or Adelta fibers. DPN terminals (some of them glutamatergic) were mainly distributed in the medial edge of the dorsal horn in the L6 spinal segment. GluR1 subunits were present in some DPN afferents, suggesting that they could be autoreceptors. DPN fibers were also present in the MPG, as were Glu terminals and GluR4 subunits. The results reveal the presence of Glu in DPN fibers and terminals and suggest that both the spinal cord and the MPG use glutamatergic transmission to control reflexive erections. PMID- 16307289 TI - [Prophylactic pancreas surgery]. AB - The goal of prophylactic surgery is to prevent malignant growth in patients with hereditary tumor predisposition. The pancreas presents as particularly challenging, due to the difficulty of operation and comparatively high risk of morbidity and even mortality. In addition, partial operative procedures and, more significantly, total resection lead to exocrine pancreas insufficiency and secondary diabetes, with grave consequences for the patient. Hereditary tumor predisposition syndromes that can result in pancreaticoduodenal endocrine tumors (PET) include multiple endocrine neoplasia type 1 syndrome and von Hippel-Lindau syndrome. As penetrance is maximally 70-80% and the 10-year survival rate over 80%, prophylactic pancreatic resection without evidence of a tumor is not indicated. However, prophylactic extension of a resection would be advised, should a PET be diagnosed. Patients predisposed to developing ductal pancreatic carcinoma (PC) are at risk of familial pancreatic cancer syndrome (FPC), hereditary pancreatitis, and other hereditary tumor predisposition syndromes such as Peutz-Jeghers syndrome and familial atypical multiple mole-melanoma syndrome. As the gene defect responsible for FPC has yet to be identified and the penetrance of PC in the other tumor predisposition syndromes is low or unknown, a prophylactic pancreatectomy based on today's knowledge is not indicated. Prophylactic extension of the resection is advisable should PC or high-grade PanIN lesions be diagnosed, as these patients often present with multifocal dysplasia and even carcinoma. PMID- 16307290 TI - A study for evaluating the effect of the deltoid-flap repair in massive rotator cuff defects. AB - The repair of massive cuff defects by direct suture often is impossible. In these cases, a repair by musculo-tendineous flaps (latissimus-dorsi, pectoralis or deltoideus) is required. It was the goal of this study to evaluate the result of delta-flap repair in case of massive cuff defects with a diameter of 5 cm or more. Between 1998 and 2000 for all patients who were suffering from a massive rotator cuff tear more than 5 cm a deltoid transfer was performed. A total of 20 patients (14 male, 6 female; age: 60.9+/-8.7 years) were available for a follow up after 47.2+/-8.0 (range, 36 to 60) month. The operation included an arthroscopic evaluation, acromioplasty with resection of the lateral clavicular end, and biceps tenodesis. The cuff defect was repaired by transfer a muscular flap from the anterior part of the deltoid (about 2 x 6 cm) into the defect. The patients subjectively rated their result-10 excellent, 9 good, and 1 poor. Preoperatively, the Constant amounted 26.3+/-5.1 points. At follow-up, the score significantly increased to 74.5+/-8.5 points. The acromiohumeral distance increased from 4.9+/-1.1 to 9.2+/-1.7 mm. In MRI examination of 11 patients all had an intact flap. Two complications (a wound hematoma and a deep infection) did not influence the result. The repair of massive rotator cuff tears by a deltoid transfer produces acceptable clinical and radiological results. PMID- 16307291 TI - Oxalate induced expression of monocyte chemoattractant protein-1 (MCP-1) in HK-2 cells involves reactive oxygen species. AB - Oxalate is a toxic end product of metabolism largely because of its propensity to crystallize and form calcium oxalate, which is insoluble at physiologic pH and often deposits at very unfortunate sites, notably the kidneys. In the current study, we investigated the oxalate-induced injury and up-regulation of monocyte chemoattractant protein-1 (MCP-1) in HK-2 cells, a proximal tubular epithelial cell line derived from normal human kidney. The cells were exposed to oxalate ions for different lengths of time. The culture media was tested for LDH release, a cell injury marker. mRNA was isolated from the cells and subjected to reverse transcriptase-polymerase chain reaction. The data showed that oxalate exposure resulted in cell injury in a time and concentration dependent manner. The MCP-1 mRNA increased following exposure to oxalate and was reduced upon treatment with free radical scavengers, catalase and superoxide dismutase. These data support the importance of reactive oxygen species in the induction of expression of MCP-1 in renal epithelial cells. To our knowledge, this is the first report of MCP-1 expression and its upregulation by oxalate exposure in HK-2 cells. PMID- 16307292 TI - Subdermal re-injection: a method to increase surgical detection of the sentinel node in breast cancer without increasing the false-negative rate. AB - PURPOSE: The aim of this study was to evaluate in breast cancer whether subdermal (SB) re-injection improves surgical detection (SD) of the sentinel node (SN) in patients with negative lymphoscintigraphy on peritumoral (PT) injection, without increasing the false-negative (FN) rate. METHODS: Group I comprised 261 patients with invasive breast cancer >3 cm and clinically negative axilla treated with primary chemotherapy. Axillary lymphadenectomy was performed in all of these patients. Group IA comprised 201 patients with PT injection, while group IB comprised 60 patients with SB injection in the tumour quadrant. Group II comprised 652 patients with breast cancer <3 cm; in 73 of these patients with negative lymphoscintigraphy, SB re-injection was performed. For lymphoscintigraphy, 37-55 MBq (99m)Tc-albumin nanocolloid in 1 ml was used for PT injection, and 18 MBq in 0.2 ml for SB injection. Five-minute images were obtained 2 h p.i. for PT injection and 20-30 min p.i. for SB injection. SD was performed 4 or 24 h p.i. Lymphoscintigraphic (LD), surgical and internal mammary (IM) detection rates were calculated. In group I, FN, negative predictive value (NPV) and accuracy (A) were calculated. Statistical analysis was performed using the chi-square test. RESULTS: In percentages, results were as follows: Group IA: SD: 84.1, FN: 13.6, NPV: 88.9, A: 78.6, IM: 14.5*. Group IB: SD: 90, FN: 0, NPV: 100, A: 90, IM: 1.7* (*p<0.025). Group II: PT injection only: LD: 82.4, SD: 94; PT injection+SB re-injection: LD: 90, SD: 98.5. SD was 97.8** in patients with positive lymphoscintigraphy and 58.5** when lymphoscintigraphy was negative (**p<0.001). CONCLUSION: For correct staging, including extra-axillary drainage, peritumoural injection should first be performed. When the SN is not visualised, and only in those cases, SB re-injection should be performed, which increases the SD rate without increasing the FN rate. PMID- 16307293 TI - Blockade of muscarinic acetylcholine receptors in the ventral tegmental area disrupts food-related learning in rats. AB - RATIONALE: Stimulation of ventral tegmental area (VTA) muscarinic acetylcholine receptors (mAChRs) is implicated in feeding. OBJECTIVE: To investigate the effects of mAChR blockade in the VTA on food-related learning. METHODS: In experiment 1, rats (N=12) were placed in chambers containing food and received microinjections of 0 or 5 microg/0.5 microl scopolamine prior to the first four feeding sessions and the alternate dose prior to the tenth feeding session. In experiment 2 (N=9), the effects of daily microinjections of scopolamine on lever pressing under a progressive ratio schedule of food reinforcement were tested. In experiment 3 (N=34), the effects of daily microinjections of scopolamine on lever pressing maintained by conditioned reward were investigated. RESULTS: In experiment 1, all rats demonstrated low consumption during session 1. However, pellet consumption for rats initially pretreated with the 0-microg dose rose to and stayed at maximal levels for the remaining sessions, even when pretreated with the 5-microg dose during the tenth session. Pellet consumption for rats initially pretreated with the 5-microg dose remained low, even for the first two sessions following the cessation of scopolamine pretreatment, and gradually rose to maximal levels by the eighth session. In experiment 2, scopolamine significantly decreased break points. In experiment 3, scopolamine failed to significantly decrease responding specifically on the lever producing the conditioned reward. CONCLUSIONS: Altogether, these data suggest that VTA mAChR stimulation is involved in feeding and food-related learning but may not be involved in responding maintained by conditioned reward. PMID- 16307294 TI - Antagonism of discriminative stimulus effects of delta(9)-THC and (R) methanandamide in rats. AB - RATIONALE: In previous drug discrimination studies we observed surmountable antagonism by Delta(9)-tetrahydrocannabinol (THC) in the presence of constant doses of SR-141716 [N-(piperidin-1-yl)-5-(4-chloro-phenyl)-1-(2,4-dichlorophenyl) 4-methyl-1H-pyrazole-3-carboxamide] (0.3 and 1 mg/kg), but there was only marginal evidence for surmountable antagonism with combinations of SR-141716 and (R)-methanandamide, a chiral analog of the endocannabioid anandamide. OBJECTIVE: Here we examine antagonism where the cannabinoid CB1 receptor agonist [Delta(9) THC and (R)-methanandamide] dose is held constant (i.e., the training dose) and the antagonist {i.e., SR-141716 and AM-251 [N-(piperidin-1-yl)-5-(4-iodophenyl)-1 (2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide; 2 ml/kg]} dose varied. We also tested the cannabinoid CB2 receptor antagonist SR-144528 {N-[(1S)-endo 1,3,3-trimethylbicyclo(2.2.1)heptan-2-yl]5-(4-chloro-3-methyl-phenyl)-1-(4 methylbenzyl)pyrazole-3-carboxamide}. METHODS: Different groups of rats were trained to discriminate between vehicle and three different doses of Delta(9)-THC (1.8, 3, and 5.6 mg/kg, presumably reflecting different efficacy demands) as well as 10 mg/kg (R)-methanandamide. Dose-generalization tests involved different doses of the cannabinoid CB1 receptor agonists. Antagonist tests varied the dose of the antagonist (range: 0.1 and 3 mg/kg for SR-141716 and AM-251, and 1 to 10 mg/kg for SR-144528). RESULTS: SR-141716 and AM-251 doses dependently blocked the agonist-induced discriminative stimulus effects. SR-141716 tended to be slightly more potent than AM-251. The effective dose 50 (ED(50)) of SR-141716 was higher in the 5.6 mg/kg Delta(9)-THC-trained group relative to the two other Delta(9) THC-trained groups. The cannabinoid CB2 receptor antagonist SR-144528 combined with the training dose of 1.8 mg/kg Delta(9)-THC, as well as when combined with the training dose of 10 mg/kg (R)-methanandamide, did not markedly change drug appropriate (agonist) responses. CONCLUSION: Data support that the discriminative stimulus effects of (R)-methanandamide and its overlap with the Delta(9)-THC cue are, indeed, CB1 receptor mediated events as revealed in antagonism tests with the selective central CB1 receptor antagonists SR-141716 and AM-251. The activation of cannabinoid CB2 receptors appears to be insignificant for these discriminations. PMID- 16307295 TI - Desipramine attenuates working memory impairments induced by partial loss of catecholamines in the rat medial prefrontal cortex. AB - RATIONALE: The density of tyrosine hydroxylase-immunoreactive (TH-IR) axons in the prefrontal cortex of schizophrenic subjects may be reduced by as much as 50% in the deep cortical layers (Am J Psychiatry 156:1580-1589, 1999). Previously, we demonstrated that approximately 60% loss of TH-IR axons in the rat medial prefrontal cortex (mPFC) decreases local basal and stress-evoked extracellular dopamine (DA) concentrations, suggesting that moderate loss of DA axons in the mPFC is sufficient to alter the neurochemical activity of the remaining DA neurons (Neuroscience 93:497-505, 1999). OBJECTIVES: To further assess the functional consequences of partial mPFC DA depletion, we examined the effects of 6-hydroxydopamine lesions of the rat mPFC on behavior in a T-maze delayed response task. We also assessed whether chronic administration of the norepinephrine (NE) uptake inhibitor, desipramine (DMI), attenuates lesion induced deficits in T-maze performance. Previous research indicates that inhibition of NE transport in the mPFC results in a concomitant increase in extracellular DA and NE. RESULTS: Moderate loss of mPFC DA and NE (approximately 50 and 10% loss, respectively) was sufficient to impair delayed-response behavior, in part due to an increase in perseverative responding. Chronic DMI treatment (3 mg/kg delivered via osmotic pumps) impaired performance of control rats but attenuated the deficits in delayed-response behavior in rats previously sustaining loss of mPFC DA and NE (approximately 75 and 35% loss, respectively). CONCLUSION: These data suggest that moderate loss of DA and NE in the prefrontal cortex is sufficient to impair cognitive function, and these behavioral effects are attenuated by inhibition of the NE transporter. PMID- 16307296 TI - Effect of noradrenergic system on the anxiolytic-like effect of DOI (5-HT2A/2C agonists) in the four-plate test. AB - RATIONALE: Selective serotonin reuptake inhibitors and serotonin and noradrenaline reuptake inhibitors demonstrated an anxiolytic-like effect in the four-plate test (FPT). (+/-)-1-(2,5-Dimethoxy-4-iodophenyl)-2-aminopropane (DOI; a 5-HT2A receptor agonist) also possessed strong anxiolytic-like effect in the same test. A 5-HT2A mechanism seems to be implicated in the mechanism of action of both antidepressants and DOI in this test. On the other hand, the alpha adrenergic ligands have also demonstrated an activity in other models of anxiety. A previous study demonstrated that the alpha2-adrenoceptor agonists abolished the anxiolytic-like effect of antidepressants. OBJECTIVES: The aim of the present study was to evaluate the role of noradrenergic system on the regulation of 5-HT2 receptors implicated in the DOI anxiolytic-like activity in the FPT. METHODS: First, the effect of noradrenergic and serotonergic lesions on DOI anxiolytic like activity was studied in the FPT. Second, the effect of co-administration of alpha-adrenoceptor ligands and DOI was evaluated in the same test. RESULTS: The noradrenergic and serotonergic lesions had no effect on DOI (1 mg/kg) anti punishment activity in the FPT. Adrafinil 0.25 and 4 mg/kg (an alpha1 adrenoceptor agonist), prazosin 0.5 and 2 mg/kg (an alpha1-adrenoceptor antagonist) and idazoxan 1 and 4 mg/kg (an alpha2-adrenoceptor antagonist) did not modify the activity of DOI. Clonidine 0.06 mg/kg, guanabenz 0.125 and 0.5 mg/kg (two alpha2-adrenoceptor agonists) and guanfacine 0.06 and 0.125 mg/kg (a specific alpha2A-adrenoceptor agonist) completely abolished DOI-induced increase in punished passages. CONCLUSION: These results indicate that the DOI seems to act on the 5-HT2 receptors post-synaptically located. The effect of DOI is regulated by the alpha2-adrenoceptors. PMID- 16307297 TI - Orthodontic treatment in children to prevent sleep-disordered breathing in adulthood. AB - The purpose of this article is to review human craniofacial growth and development, especially the growth of the mandible, to clarify the relationship between obstructive sleep apnea (OSA) syndrome and craniofacial abnormality, and finally, to propose the hypothesis that negative pressure produced in the chest of the OSA child inhibits the growth of the mandible. Recently, the development of diagnosis and treatment of OSA syndrome has progressed rapidly; however, the prevention of OSA syndrome was merely seen. Craniofacial abnormality is reported as one of the causes of OSA syndrome. If craniofacial abnormality is determined only by genetics, it is difficult to manage the craniofacial skeleton to prevent OSA syndrome. The role of epigenetic factors on craniofacial growth and development is still controversial. However, if we stand on the functional matrix hypothesis, we can manage not only growth of the mandible but also the craniofacial skeleton as a whole. The author proposes the hypothesis that the negative pressure produced in the chest prohibits the growth of the mandible even if the patients have a capacity for growth and development; therefore, if this negative pressure disappears because of the removal of the tonsil and/or adenoids or by an orthodontic treatment to make a patency of the airway, the mandible may grow normally, and we can prevent or reduce a number of OSA syndromes in the future. PMID- 16307299 TI - Academy of Dental Sleep Medicine--position statement. Dental sleep medicine and portable monitoring, August 2005. PMID- 16307300 TI - Carcinomatous meningitis from urachal carcinoma: the first reported case. AB - Carcinomatous meningitis (CM) occurs in less than 10% of cancer patients. Although patients frequently present with a focal complaint, multifocal signs are often found following careful neurological examination. The gold standard for diagnosis remains the demonstration of neoplastic cells in the cerebrospinal fluid. Despite the discouraging prognosis, palliative treatment may improve quality of life and lengthen lifespan. We report a patient with known primary carcinoma of the urachus who presented with headaches, nausea, vomiting and ataxia 1 week following resection of a nodular arachnoidal metastasis (indenting the cerebellum). Lumbar cerebrospinal fluid subsequently confirmed carcinomatous meningitis. This is the first reported case of carcinomatous meningitis resulting from metastatic urachal carcinoma. PMID- 16307301 TI - Synthesis of the sulfur amino acids: cysteine and methionine. AB - This review will assess new features reported for the molecular and biochemical aspects of cysteine and methionine biosynthesis in Arabidopsis thaliana with regards to early published data from other taxa including crop plants and bacteria (Escherichia coli as a model). By contrast to bacteria and fungi, plant cells present a complex organization, in which the sulfur network takes place in multiple sites. Particularly, the impact of sulfur amino-acid biosynthesis compartmentalization will be addressed in respect to localization of sulfur reduction. To this end, the review will focus on regulation of sulfate reduction by synthesis of cysteine through the cysteine synthase complex and the synthesis of methionine and its derivatives. Finally, regulatory aspects of sulfur amino acid biosynthesis will be explored with regards to interlacing processes such as photosynthesis, carbon and nitrogen assimilation. PMID- 16307302 TI - Expression profiling of metabolic genes in response to methyl jasmonate reveals regulation of genes of primary and secondary sulfur-related pathways in Arabidopsis thaliana. AB - The treatment of Arabidopsis thaliana with methyl jasmonate was used to investigate the reaction of 2467 selected genes of primary and secondary metabolism by macroarray hybridization. Hierarchical cluster analysis allowed distinctions to be made between diurnally and methyl jasmonate regulated genes in a time course from 30 min to 24 h. 97 and 64 genes were identified that were up- or down-regulated more than 2-fold by methyl jasmonate, respectively. These genes belong to 18 functional categories of which sulfur-related genes were by far strongest affected. Gene expression and metabolite patterns of sulfur metabolism were analysed in detail, since numerous defense compounds contain oxidized or reduced sulfur. Genes encoding key reactions of sulfate reduction as well as of cysteine, methionine and glutathione synthesis were rapidly up-regulated, but none of the known sulfur-deficiency induced sulfate transporter genes. In addition, increased expression of genes of sulfur-rich defense proteins and of enzymes involved in glucosinolate metabolism was observed. In contrast, profiling of primary and secondary sulfur metabolites revealed only an increase in the indole glucosinolate glucobrassicin upon methyl jasmonate treatment. The observed rapid mRNA changes were thus regulated by a signal independent of the known sulfur deficiency response. These results document for the first time how comprehensively the regulation of sulfur-related genes and plant defense are connected. This interaction is discussed as a new approach to differentiate between supply- and demand-driven regulation of the sulfate assimilation pathway. PMID- 16307303 TI - Chloroplast sulfate transport in green algae--genes, proteins and effects. AB - This review summarizes evidence at the molecular genetic, protein and regulatory levels concerning the existence and function of a putative ABC-type chloroplast envelope-localized sulfate transporter in the model unicellular green alga Chlamydomonas reinhardtii. From the four nuclear genes encoding this sulfate permease holocomplex, two are coding for chloroplast envelope-targeted transmembrane proteins (SulP and SulP2), a chloroplast stroma-targeted ATP binding protein (Sabc) and a substrate (sulfate)-binding protein (Sbp) that is localized on the cytosolic side of the chloroplast envelope. The sulfate permease holocomplex is postulated to consist of a SulP-SulP2 chloroplast envelope transmembrane heterodimer, flanked by the Sabc and the Sbp proteins on the stroma side and the cytosolic side of the inner envelope, respectively. The mature SulP and SulP2 proteins contain seven transmembrane domains and one or two large hydrophilic loops, which are oriented toward the cytosol. The corresponding prokaryotic-origin genes (SulP and SulP2) probably migrated from the chloroplast to the nuclear genome during the evolution of Chlamydomonas reinhardtii. These genes, or any of its homologues, have not been retained in vascular plants, e.g. Arabidopsis thaliana, although they are encountered in the chloroplast genome of a liverwort (Marchantia polymorpha). The function of the SulP protein was probed in antisense transformants of C. reinhardtii having lower expression levels of the SulP gene. Results showed that cellular sulfate uptake capacity was lowered as a consequence of attenuated SulP gene expression in the cell, directly affecting rates of de novo protein biosynthesis in the chloroplast. The antisense transformants exhibited phenotypes of sulfate-deprived cells, displaying slow rates of light-saturated oxygen evolution, low levels of Rubisco in the chloroplast and low steady-state levels of the Photosystem II D1 reaction center protein. The role of the chloroplast sulfate transport in the uptake and assimilation of sulfate in Chlamydomonas reinhardtii is discussed along with its impact on the repair of Photosystem II from a frequently occurring photo oxidative damage and H2-evolution related metabolism in this green alga. PMID- 16307304 TI - Chemical modification studies of tryptophan, arginine and lysine residues in maize chloroplast ferredoxin:sulfite oxidoreductase. AB - The ferredoxin-dependent sulfite reductase from maize was treated, in separate experiments, with three different covalent modifiers of specific amino acid side chains. Treatment with the tryptophan-modifying reagent, N-bromosuccinimide (NBS), resulted in a loss of enzymatic activity with both the physiological donor for the enzyme, reduced ferredoxin, and with reduced methyl viologen, a non physiological electron donor. Formation of the 1:1 ferredoxin/sulfite reductase complex prior to treating the enzyme with NBS completely protected the enzyme against the loss of both activities. Neither the secondary structure, nor the oxidation-reduction midpoint potential (Em) values of the siroheme and [4Fe-4S] cluster prosthetic groups of sulfite reductase, nor the binding affinity of the enzyme for ferredoxin were affected by NBS treatment. Treatment of sulfite reductase with the lysine-modifying reagent, N-acetylsuccinimide, inhibited the ferredoxin-linked activity of the enzyme without inhibiting the methyl viologen linked activity. Complex formation with ferredoxin protects the enzyme against the inhibition of ferredoxin-linked activity produced by treatment with N acetylsuccinimide. Treatment of sulfite reductase with N-acetylsuccinimide also decreased the binding affinity of the enzyme for ferredoxin. Treatment of sulfite reductase with the arginine-modifying reagent, phenylglyoxal, inhibited both the ferredoxin-linked and methyl viologen-linked activities of the enzyme but had a significantly greater effect on the ferredoxin-dependent activity than on the reduced methyl viologen-linked activity. The effects of these three inhibitory treatments are consistent with a possible role for a tryptophan residue the catalytic mechanism of sulfite reductase and for lysine and arginine residues at the ferredoxin-binding site of the enzyme. PMID- 16307305 TI - Selenium uptake, translocation, assimilation and metabolic fate in plants. AB - The chemical and physical resemblance between selenium (Se) and sulfur (S) establishes that both these elements share common metabolic pathways in plants. The presence of isologous Se and S compounds indicates that these elements compete in biochemical processes that affect uptake, translocation and assimilation throughout plant development. Yet, minor but crucial differences in reactivity and other metabolic interactions infer that some biochemical processes involving Se may be excluded from those relating to S. This review examines the current understanding of physiological and biochemical relationships between S and Se metabolism by highlighting their similarities and differences in relation to uptake, transport and assimilation pathways as observed in Se hyperaccumulator and non-accumulator plant species. The exploitation of genetic resources used in bioengineering strategies of plants is illuminating the function of sulfate transporters and key enzymes of the S assimilatory pathway in relation to Se accumulation and final metabolic fate. These strategies are providing the basic framework by which to resolve questions relating to the essentiality of Se in plants and the mechanisms utilized by Se hyperaccumulators to circumvent toxicity. In addition, such approaches may assist in the future application of genetically engineered Se accumulating plants for environmental renewal and human health objectives. PMID- 16307306 TI - Sulfite oxidation in plant peroxisomes. AB - For a long time, occurrence and nature of sulfite oxidase activity in higher plants were controversially discussed. During primary sulfate assimilation in the chloroplast, sulfate is reduced via sulfite to organic sulfide, which is essential for cysteine biosynthesis. However, it has also been reported that sulfite can be oxidized back to sulfate, e.g. when plants were subjected to SO2 gas. Recently, work from our laboratory has identified the sulfite oxidase as the fourth member of molybdenum-enzymes in plants. Here we discuss how nature separates the two counteracting pathways--sulfate assimilation and sulfite detoxification--into two different cell organelles and we will also discuss how these two processes are coregulated. PMID- 16307307 TI - Thioredoxins in Arabidopsis and other plants. AB - Regulation of disulfide dithiol exchange has become increasingly important in our knowledge of plant life. Initially discovered as regulators of light-dependent malate biosynthesis in the chloroplast, plant thioredoxins are now implicated in a large panel of reactions related to metabolism, defense and development. In this review we describe the numerous thioredoxin types encoded by the Arabidopsis genome, and provide evidence that they are present in all higher plants. Some results suggest cross-talk between thioredoxins and glutaredoxins, the second family of disulfide dithiol reductase. The development of proteomics in plants revealed an unexpectedly large number of putative target proteins for thioredoxins and glutaredoxins. Nevertheless, we are far from a clear understanding of the actual function of each thioredoxin in planta. Although hampered by functional redundancies between genes, genetic approaches are probably unavoidable to define which thioredoxin interacts with which target protein and evaluate the physiological consequences. PMID- 16307308 TI - Insights into the acclimation of Chlamydomonas reinhardtii to sulfur deprivation. AB - During sulfur deprivation, the photosynthetic green alga Chlamydomonas reinhardtii develops a high-affinity sulfate uptake system and increases the expression of genes encoding proteins involved in sulfur assimilation. Although two regulatory elements, SAC1 and SAC3, have been shown to be required for normal acclimation of C. reinhardtii to sulfur deprivation, a number of other regulatory elements appear to also be involved. The molecular mechanisms by which these regulatory elements function are largely unknown. This manuscript presents our current knowledge of sulfur deprivation responses and the regulation of these responses in C. reinhardtii. In addition, we present preliminary results of a sub saturation screen for novel sulfur acclimation mutants of C. reinhardtii. A speculative model, incorporating the activities of established regulatory elements with putative novel components of the signal transduction pathway(s) is discussed. PMID- 16307309 TI - Sulfate assimilation and glutathione synthesis in C4 plants. AB - Sulfate assimilation and glutathione synthesis were traditionally believed to be differentially compartmentalised in C4 plants with the synthesis of cysteine and glutathione restricted to bundle sheath and mesophyll cells, respectively. Recent studies, however, showed that although ATP sulfurylase and adenosine 5' phosphosulfate reductase, the key enzymes of sulfate assimilation, are localised exclusively in bundle sheath in maize and other C4 monocot species, this is not true for the dicot C4 species of Flaveria. On the other hand, enzymes of glutathione biosynthesis were demonstrated to be active in both types of maize cells. Therefore, in this review the recent findings on compartmentation of sulfate assimilation and glutathione metabolism in C4 plants will be summarised and the consequences for our understanding of sulfate metabolism and C4 photosynthesis will be discussed. PMID- 16307311 TI - Validated ligand mapping of ACE active site. AB - Crystal structures of angiotensin-converting enzyme (ACE) complexed with three inhibitors (lisinopril, captopril, enalapril) provided experimental data for testing the validity of a prior active site model predicting the bound conformation of the inhibitors. The ACE active site model - predicted over 18 years ago using a series of potent ACE inhibitors of diverse chemical structure - was recreated using published data and commercial software. Comparison between the predicted structures of the three inhibitors bound to the active site of ACE and those determined experimentally yielded root mean square deviation (RMSD) values of 0.43-0.81 A, among the distances defining the active site map. The bound conformations of the chemically relevant atoms were accurately deduced from the geometry of ligands, applying the assumption that the geometry of the active site groups responsible for binding and catalysis of amide hydrolysis was constrained. The mapping of bound inhibitors at the ACE active site was validated for known experimental compounds, so that the constrained conformational search methodology may be applied with confidence when no experimentally determined structure of the enzyme yet exists, but potent, diverse inhibitors are available. PMID- 16307310 TI - Sulfur and primary production in aquatic environments: an ecological perspective. AB - Sulfur is one of the critical elements in living matter, as it participates in several structural, metabolic and catalytic activities. Photosynthesis is an important process that entails the use of sulfur during both the light and carbon reactions. Nearly half of global photosynthetic carbon fixation is carried out by phytoplankton in the aquatic environment. Aquatic environments are very different from one another with respect to sulfur content: while in the oceans sulfate concentration is constantly high, freshwaters are characterized by daily and seasonal variations and by a wide range of sulfur concentration. The strategies that algal cells adopt for energy and resource allocation often reflect these differences. In the oceans, the amount and chemical form of sulfur has changed substantially during the course of the Earth's history; it is possible that sulfur availability played a role in the evolution of marine phytoplankton communities and it may continue to have appreciable effects on global biogeochemistry and ecology. Phytoplankton is also the main biogenic source of sulfur; sulfur can be released into the atmosphere by algal cells as dimethylsulfide, with possibly important repercussions on global climate. These and related matters are discussed in this review. PMID- 16307312 TI - Functional assays of HLA A2-restricted epitope variant of latent membrane protein 1 (LMP-1) of Epstein-Barr virus in nasopharyngeal carcinoma of Southern China and Taiwan. AB - Human leukocyte antigen (HLA) A2 was consistently associated with increased risk for nasopharyngeal carcinoma (NPC) in Chinese populations. Previously we have reported that an Epstein-Barr virus (EBV) strain carrying an HLA A2-restricted epitope variant of LMP-1 is prevalent in NPC in southern China and Taiwan (Lin et al., J. Gen. Virol. 85: 2023-2034, 2004). The variant has mutation selectively involved one of the two anchor residues in position 2 (125 L-->F) and an additional mutation in position 5 (129 M-->I). Functional assays of the epitope variant were carried out in the present work. The stabilization assay on T2 cells indicated that the variant peptide YFL (YFLEILWRL) prevalent in NPC binds to HLA A2 molecules less efficiently than the prototype peptide YLL (YLLEMLWRL). A dose dependent binding of the HLA A2 molecules with added peptides was observed. In ex vivo cytotoxic T lymphocyte (CTL) assays with CD8-enriched effectors from A2 positive donors revealed that the YLL-specific CTL was able to lyse EBV-infected B cells expressing HLA A2, whereas the CTL recognition was abrogated with the peptide YFL. Cytokine (IFN-gamma) responses, measured both by intracytoplasmic staining and ELISPOT assays after peptide stimulation, also indicated that the variant epitope peptide failed to give an IFN-gamma response. The IFN-gamma response was almost entirely restricted to those tetramer-positive cells. These results show that EBV isolates from NPC of southern China and Taiwan is dominated by an HLA A2-restricted 'epitope-loss variants' of LMP-1, which would allow the virus to resist immune recognition and may in part contribute to the prevalence of NPC in these populations. PMID- 16307313 TI - Three-dimensional recovery time dispersion map by 64-channel magnetocardiography may demonstrate the location of a myocardial injury and heterogeneity of repolarization. AB - BACKGROUND: QT dispersion reveals heterogeneities in the repolarization time in the three-dimensional (3D) structure of the ventricular myocardium. In this study, we report on a 3D function map of recovery time (RT) dispersions as measured by 64-channel magnetocardiography (MCG). METHODS: MCG were simultaneously recorded in 29 controls and 21 patients with previous myocardial infarction (MI). The 3D current density was calculated from 64-channel MCG data in the Bz component using a space filter. The heart outline, reconstructed from the integrated the current density, revealed both the atrium and ventricle. The RT for the intervals between QRS onset and the time of the maximum dT/dt of T wave, and the peak to the end of the T wave (T(peak)-negative dT/dt) were automatically measured by means of a computer from 3D MCG data. The corrected RT (RTc) and corrected T(peak)-negative dT/dt were then calculated using Bazett's formula. The 3D RTc and the corrected T(peak)-negative dT/dt dispersion map were superimposed on the heart outline generated by MCG. RESULTS: The RTc was significantly longer for the MI group than in the control group (67+/-25 ms1/2 vs. 16+/-6 ms1/2) (p<0.0001). The corrected T(peak)-negative dT/dt dispersions in each patient was also significantly longer for the MI group than in the control group (35+/-27 ms1/2 vs. 10+/-5 ms1/2) (p<0.0001). Furthermore, the 3D RTc and T(peak)-negative dT/dt dispersion maps corresponded with the space location of MI, as defined by Tc-99m tetrofosmin myocardial imaging CONCLUSIONS: 3D RTc and T(peak)-negative dT/dt dispersion maps in the ST segment, obtained by 64-channel MCG may be used demonstrate the location of a myocardial injury and heterogeneities of repolarization. PMID- 16307314 TI - Histogenesis of human renal cell carcinoma by using electron microscopy and immunohistochemical techniques. AB - Electron microscopy and immunohistochemical techniques are powerful tools for the determination of tissue origin. Both techniques have been used in the current experiment for histogenesis of renal cell carcinoma. Fifty kidney tumors were subjected to immunohistochemical detection for intermediate filaments cytokeratin and vimentin, which are normally expressed in epithelial tissue and mesenchymal tissues, respectively. Twenty cases of the above were examined by electron microscopy for detection of ultrastructure features. From each kidney, two specimens were taken, one from the diseased area and another far from it to serve as a control. Immunohistochemical study revealed in cases of renal cell carcinoma, cytokeratin and vimentin were expressed alone in 44% of cases, and 40% of cases, respectively. Twelve percent of cases were coexpressed with both cytokeratin and vimentin. Electron microscopic study of diseased specimens revealed the expression of desmosomes which was observed in almost all tumor specimens. The expression of the vimentin in some cases either alone or with cytokeratin was interpreted as a change in the characters of some tumor cells which indicates the need for additional techniques in such cases to get the proper interpretation. The prevalence of the expression of cytokeratin and the persistence existence of desmosomes indicate the epithelial origin of the tumor. This data is very beneficial for determination of line of therapy and follow up of the patients. The results confirm the power of combined use of both immunohistochemistry and electron microscopy in the field of histogenesis. PMID- 16307315 TI - Prognostic factors effecting on recurrence of urinary stone disease: a multivariate analysis of everyday patient parameters. AB - INTRODUCTION: Since comprehensive urometabolic analyses are currently more preferred for the patients with recurrent stones and with high risk of stone recurrence, we have tried to determine simple patient data increasing stone recurrence to limit sophisticated analyses to certain cases at least in particularly deprivation districts. MATERIALS AND METHODS: For the study 173 with first-time or recurrent urinary stone were taken. After stone treatment, various empirical metaphylaxis methods were counselled to the patients. The parameters evaluated were (a) age at onset of the disease, (b) gender, (c) urinary pH and (d) specific gravity, (e) serum calcium and (f) uric acid, (g) stone burden, (h) side, and (i) location, (j) treatment modality and (k) recurrence history. New stone formation or growing of the existing stone was considered as stone recurrence. In statistics, independent samples t, chi-square test, Kaplan-Meier, Log rank and Cox regression tests were used. RESULTS: The mean age was 35 years. The male to female ratio was 88/85. Recurrence occurred in 49 (28%) cases at a mean of 30 months. Stone burden was significantly larger in patients with recurrent stone. In subjects treated with open surgery and with previous recurrence history, stone recurrence rate was significantly higher. In survival analyses, higher serum calcium level, larger stone burden, renal stones and previous recurrence influenced stone prognosis poorly. However in multivariate analysis, none of them was the most significant independent factor. CONCLUSIONS: According to our study, detailed urometabolic analyses may be somewhat reserved for the patients with ordinary factors increasing risk of stone recurrence including relatively high serum calcium level, large stone burden, upper urinary stone, history of recurrence and open surgery. As a result, it has been thought that the stone risk evaluation guiding detailed laboratory examination may be partially performed with first-line clinical data under limited conditions. PMID- 16307316 TI - Renal calculi in a patient with Erdheim-Chester disease. AB - Erdheim-Chester Disease (ECD) is a non-inherited multifocal lipid storing histiocytosis. It is a rare disease characterised by lipid-laden monocyte infiltration of long bones causing cortical sclerosis and characteristic X-ray appearances. It also involves extraskeletal tissue in up to 50% of cases including retroperitoneal and renal infiltration. We report a patient with long standing ECD with widespread extraskeletal involvement, including significant renal infiltration, presenting with left hydronephrosis secondery obstruction from a proximal ureteric calculas. PMID- 16307317 TI - Giant renal hydatid cyst resembling a simple cyst: an intraoperative diagnosis. AB - An interesting case of a giant renal hydatid cyst is presented. The big cystic mass detected at ultrasonography (US) and computerized tomography (CT) in a ten year-old girl looked like a simple cyst. No germinative membrane or any other radiological sign of a hydatid cyst was present. The operation was planned for a simple cyst; but was altered, after an intraoperative diagnostic needle sampling brought out the characteristic fluid of hydatid cyst. We came to the conclusion that hydatid cystic masses may not always present with their characteristic radiological findings; and that extreme caution should be practiced by the radiologist and the surgeon in order to prevent iatrogenic echinococcal dissemination. PMID- 16307318 TI - A new percutaneous approach for the treatment of hydatid cyst of the kidney: long term follow-up. AB - Renal hydatid disease is an uncommon benign parasitic infestation compared to liver hydatid disease. Although the treatment of hydatid cysts depends on surgery which is related with high morbidity and an overall local recurrence rates, the following case reports describing two patients with renal hydatid cysts were successfully treated with this new percutaneous method. PMID- 16307319 TI - Collecting duct carcinoma of the renal medulla presenting with paraplegia. AB - We report an interesting case of a patient with collecting duct carcinoma arising from the left kidney who presented with paraplegia secondary to metastases. The diagnosis was based on CT and histology. To our knowledge this is the first case of collecting duct carcinoma to present with paraplegia. The literature review also highlights the rarity of this disease with less than a hundred cases reported to date and the aggressive nature and poor prognosis despite prompt interventions. PMID- 16307320 TI - A case of pyonephrosis secondary to ureteral stent calculus. AB - Ureteric stents, a good solution to many urologic problems, can lead to significant morbidity if left in situ longer than required. Our case is that of an elderly male with a history of intractable hypertension presenting with urosepsis, anemia, diabetes and chronic renal failure. His work-up revealed a stent in the right kidney with secondary staghorn calcification in the renal pelvis and a large vesical calculus. He had apparently undergone stent placement 12 years previously, but was lost to follow-up due to relocation. A nuclear scan revealed a complete loss of renal function. Cystoscopic stent removal was futile, so he underwent an elective right subcapsular nephrectomy. The specimen revealed pyonephrosis with loculations of pus. Postoperatively his course was uneventful, with the hypertension resolving in 4 weeks. PMID- 16307321 TI - A day case technique for administration of intradetrusor Botulinum toxin B under sedo-analgesia in neuropathic and non-neuropathic detrusor overactivity: Endoscopic Neurostabilisation (ENS). PMID- 16307322 TI - Congenital vesical diverticulum in a 38-year-old female. AB - A 38-year-old female with a history of psychiatric illness presents with irritative urological symptoms and is diagnosed as having a large congenital vesical diverticulum (Hutch diverticulum). An overview of the aetiology, classification and complications of diverticula together with indications for and management of bladder diverticula is discussed. This is the first documented case of a very large symptomatic congenital diverticulum in a female presenting in the third decade of life and emphasizes the importance of ultrasound in irritative urological symptoms resistant to medication. PMID- 16307323 TI - Nephrogenic adenoma of the urinary bladder. AB - INTRODUCTION: The nephrogenic adenoma (NA) is a benign metaplastic lesion of the urothelium and is attributed to chronic irritation of the mucosa, by injury, infection, stone disease or intravesical instrumentations. We present our experience on this morbid entity, its clinical appearance in the urinary bladder, its frequency and relapses. Furthermore we reviewed the related recent literature and focused on its potential to neoplastic degeneration and the value of the new diagnostic modalities. PATIENTS AND METHODS: Four patients with NA of the urinary bladder are presented. The papilloid or polypoid formations observed by the cystoscopy were identified after the TUR, as NA of the urinary bladder. Their mean follow-up was 3.5 years. RESULTS: Remission of the symptoms was observed after TUR in all patients. Three out of four patients presented 1-7 relapses, while in one case, after seven NA relapses, a urothelial carcinoma of the bladder was diagnosed. CONCLUSIONS: Unlike histological features, the clinical - endoscopic characteristics of NA are non-specific. Even if it is not definitely considered like a premalignant condition, NA has to be followed up frequently and long lasting, because of its high recurrence rate. The combination of Cytology, Flow cytometry, DNA image analysis and Fluorescence in situ hybridization of bladder washings or voided urine, are of high value in monitoring NA of the urothelium. PMID- 16307324 TI - Xanthoma of the urinary bladder. AB - Xanthomas of the urinary bladder are rare lesions of reactive nature. They present as yellow-white mucosal plaques that may be mistaken for a neoplastic process. A biopsy displays stromal accumulation of lipid macrophages without accompanying inflammatory component. This article presents a 78-year-old woman who had a tumour-suspect xanthoma of the urinary bladder occurring subsequent to several resections of non-invasive carcinomas. PMID- 16307325 TI - The prognostic importance of e-cadherin and p53 gene expression in transitional bladder carcinoma patients. AB - OBJECTIVE: The clinical course of transitional cell carcinoma is highly variable. The determination of sensitive prognostic factors for transitional cell carcinoma is very important. Therefore e-cadherin and p53 immunohistochemical activity can be used with other prognostic factors. METHODS: The study comprised with 61 (4 women and 57 men) selected patients who had transitional cell carcinoma. Paraffin embedded tissue sections were investigated immunohistochemically for e-cadherin normal staining and p53 over expression. RESULTS: It is seen that when grade and stages of illness increased normal staining of e-cadherin decreased and p53 over expressed. Abnormal e-cadherin was significantly associated with disease recurrence (P < 0.001), disease progression (P < 0.001) and bladder specific survival. p53 differentiation was not significant for disease recurrence (P > 0.05) inverse to prognosis of illness. Transurethral resectomy and BCG treatments were not effected e-cadherin and p53 activity within the groups statistically. CONCLUSION: Significant differences can be helpful to investigate patients more detailed pathologically. These expression rates in different type of transitional cell carcinoma patients may represent a biologically more aggressive cancer, requiring early definitive therapy. This hypothesis should be evaluated in larger studies and prospective clinical trials. PMID- 16307326 TI - Knotting of a ureteric stent in a child. AB - Knotting of ureteric stents is an unusual complication of their use and has not previously been reported in a child. We present a case of a 4-year-old that developed symptomatic hydronephroureter following removal of a multilength stent, which had knotted at the proximal loop. The child made a full recovery but knotting must be considered if resistance is encountered during stent removal. PMID- 16307327 TI - Localized primary amyloidosis of the prostate, bladder and ureters. AB - Primary amyloidosis of lower urinary tract is a rare entity and is usually localized to a single site. Its clinical features, and cystoscopic and radiological findings are indistinguishable from neoplastic or inflammatory lesions. We report an unusual case of amyloidosis involving multiple sites (prostatic stroma, trigone and lower ureters) in the lower urinary tract. MRI findings of bladder amyloid, which could be used to suspect this condition, are also described. PMID- 16307328 TI - Continent urinary diversion in patients with pelvic irradiation: an alternative utilizing transverse colon reservoir. AB - The use of radiation therapy in pelvic malignancies increases the risk of urinary complications, sometimes being necessary urinary diversion. The risk of utilizing previously irradiated bowel should be avoided. The use of transverse colon is a safe and effective alternative. We present a heterotopic continent colonic reservoir with an easily catheterizable conduit. PMID- 16307329 TI - Thermo-expandable intra-prostatic stent in the treatment of acute urinary retention in elderly patients with significant co-morbidities. AB - We assess the use of thermo-expandable intra-prostatic stent (Memokath, Engineers and Doctors A/S, Denmark) for the treatment of acute urinary retention (AUR) in men with significant co-morbidities for transurethral resection of prostate (TURP). We evaluate the pre- and post-operative complications, duration of stents in-situ and patients quality of life after the stent insertion. Patients with significant co-morbidities presenting with AUR were selected, who were unfit for TURP. The co-morbidities included ischaemic heart disease, congestive heart failure, and chronic obstructive pulmonary disease. The exclusion criteria were bladder tumour and atonic bladder. The Memokath stents were inserted using a flexible cystoscope under local anaesthesia. The patients were followed up at 3 and 6 months after the procedure and the ones who remained alive were asked to complete self-administered questionnaires and IPSS scores. Fifteen men with acute urinary retention were recruited for stent insertion with the mean age of 87 years. No peri-operative complications were recorded. Three patients died after the insertion with functional Memokath in-situ. Nine patients had good functioning stents post-operatively, and remain catheter free up to 30 months after the procedure. The mean duration of stent life was 18 months. Three long term complications were detected, including stent migration and prostate overgrowth. The Memokath is a good option for frail elderly patients presenting with AUR. The procedure is safe and has minimal long term complications. The stent also provides a sustained good quality of life for patients and avoids the necessity of long term catheterisation. PMID- 16307330 TI - Unusual cause of obstructive uropathy. AB - We report a case involving a 55-year-old healthy man that presented with urinary retention and a lower abdominal mass. His evaluation revealed a large mucocele arising from the appendix that caused severe bilateral ureteral obstruction and renal failure. Appendiceal mucoceles are rare and insidious entities that can present with urologic manifestation. Despite their rarity, this case highlights the importance of including mucoceles in the differential diagnosis in cases involving ureteral obstruction caused by external compression. PMID- 16307331 TI - Long term follow up of men with Alfuzosin who voided successfully following acute urinary retention*. AB - INTRODUCTION: Acute urinary retention (AUR) is one of the commonest causes of admission in urology ward and successful voiding with alpha-blockers has been reported. However, long-term efficacy of Alfuzosin, following an episode of AUR is lacking. This is a continuation of our earlier reported study. We report the results of a 4 year follow-up on patients who were on Alfuzosin SR 5 mg BD, following resumption of voiding after an episode of AUR. PATIENTS AND METHODS: A total of 33 patients voided successfully following AUR in our original study. These patients continued on Alfuzosin SR 5 mg BD and were assessed at 2 and 4 years. Symptomatic assessment was performed with IPSS and QOL symptom score and objective assessment was with urinary flow rate and post void residual volume. Patients who continued to deteriorate symptomatically and objectively or developed further AUR were listed for surgery. RESULTS: Out of 33 patients, 28 patients were followed up at 2 years (three patients died due to various medical reasons and two did not attend for follow-up). Nineteen patients (68%) underwent transurethral resection of prostate (TURP) for severe lower urinary tract symptom (LUTS) . The mean peak flow rate at 2 years was 8.4 ml/s and the mean residual volume was 112 ml. Ten patients attended for follow up at 4 years. The mean flow rate was 5.17 ml/s and the mean post-void residual volume was 101 ml. Four patients underwent TURP for severe outflow symptoms. At 4 years follow up 24 out of 30 patients (80%) on Alfuzosin needed TURP. CONCLUSIONS: These data do not support the long term use of alpha-blockers in patients who voided successfully after acute urinary retention. PMID- 16307332 TI - The value of PSA, free-to-total PSA ratio and PSA density in the prediction of pathologic stage for clinically localized prostate cancer. AB - OBJECTIVE: The ability of prostate-specific antigen (PSA), free/total PSA and PSA density to predict the pathologic stage in prostate cancer has not been clear yet. In this study, we evaluated the value of PSA subgroups in the prediction of pathologic stage after radical prostatectomy. METHODS: A total of 42 subjects 55 78-years-old who underwent radical retropubic prostatectomy were included in the study. Preoperative PSA, free/total PSA and PSA density (PSAD) values were compared according to the pathologic stages of radical prostatectomy specimens. Receiver operating characteristics (ROC) curves were measured for each parameter. RESULTS: The clinical stage that was estimated for all patients was between T1N0M0 and T2bN0M0. Pathologic examination revealed organ-confined disease in 18 patients. The area under curve (AUC) for organ confinement was 0.553 for PSA, 0.446 for free/total PSA ratio and 0.706 for PSAD. Cut-off values providing the best sensitivity and specificity in ROC analysis for PSA, free/total PSA and PSAD were 7.1, 0.15, and 0.17, respectively (likelihood ratio: 0.9, 1 and 2). The positive predictive values at these cut-off values were 0.54, 0.56, and 0.70, respectively. Only PSAD cut-off values was found statistically borderline significant for predicting organ-confined disease. CONCLUSION: While PSAD is more helpful than PSA and free/total PSA ratio for prediction of organ-confined disease, none of these parameters are significant predictor of pathologic stage for clinically localized prostate cancer. PMID- 16307333 TI - Microscopic polyangiitis complicated by the development of prostate cancer and flutamide-induced hepatitis. AB - We report a case of a 65-year-old man with microscopic polyangiitis who developed prostate cancer and gastric adenocarcinoma after prolonged oral use of cyclophosphamide. Acute hepatitis with jaundice and marked increase in aminotranferases occurred after 6 months of flutamide treatment for metastatic prostate carcinoma. It is suggested that patients with vasculitis or other autoimmune disorders should avoid prolonged use of cyclophosphamide and other cytotoxic drugs in order to minimize long-term adverse effects, of which the risk of cancer is by far the most important. In patients on flutamide treatment, careful monitoring of flutamide administration with repeated liver function tests should be undertaken, and the drug must be immediately discontinued in patients with abnormal results to avoid progression of liver injury. PMID- 16307334 TI - A new option for the management of urethral trauma: primary reconstruction of posterior urethral disruption with a buccal mucosa transplant. AB - Up to now, the management of traumatic posterior urethral disruption ranges from primary realignment to delayed urethroplasty. However, we reconstructed the membranous part of the urethra with an onlay graft of buccal mucosa after traumatic complete disruption as a first line therapy. After 7 months followup, the clinical outcome is very good. Because primary reconstruction of the urethra with a buccal mucosa graft after traumatic disruption has not yet been reported, the question arises whether this technique should be routinely included as an option for primary urethral reconstruction after trauma of the posterior urethra. PMID- 16307335 TI - Urinary retention, erectile dysfunction and penile anaesthesia after circumcision: a mixed dissociative (conversion) disorder. AB - We report on the long-term followup of a mixed dissociative (conversion) disorder after circumcision in childhood. PMID- 16307336 TI - Giant spermatocytic seminoma with massive hemorrhage into accompanying hydrocele: case report. AB - A 59-year-old man was admitted with giant tumor of testis and massive hemorrhage into accompanying hydrocele. His clinical status had become worse quickly. The emergency resection of tumor with hematocele was performed. The diagnosis was made upon histological examination of the postoperative tissue and was confirmed by immunohistochemistry. Spermatocytic seminoma was recognized. The surgical specimen comprised a tumor (26 cm x 28 cm x 17 cm) in size, weighed 2150 g. Tumor with hematocele achieved about 40 cm in diameter. PMID- 16307337 TI - Non seminomatous germ cell tumour of a solitary testis in a 70-year-old man: a rare case. AB - We report a rare case of a 70-year-old patient who presented with undifferentiated malignant teratoma arising from a solitary testis. The literature review highlights the rarity of such tumour in this age group, and this may represent the oldest reported patient with non seminomatous germ cell tumour of testicle. PMID- 16307338 TI - Sperm recovery prediction in azoospermic patients using Doppler ultrasonography. AB - PURPOSE: To evaluate power Doppler ultrasonography to predict sperm recovery in azoospermic patients. METHODS: Color Doppler and power Doppler ultrasonography of testis were performed in 38 patients before testicular sperm extraction. Analysis of blood flow included the pulsatility and resistance index of intratesticular vessels and testicular artery, and power Doppler of testis. The results of power Doppler of testis were classified into three categories: 0, no vessels found; 1, one to three vessels; 2, more than three vessels found. RESULTS: Power Doppler of both testis showed a significant difference between obstructive azoospermia and non-obstructive azoospermia (Fisher's exact test - P = 0.02), and between the groups with and without sperm recovery (Fisher's exact test - P = 0.001). Doppler indices of intratesticular vessels and testicular artery were similar between the groups. CONCLUSIONS: Testicular Power Doppler assessment showed that patients with obstructive azoospermia have better blood flow than patients with non obstructive azoospermia, and power Doppler is able to predict sperm recovery in azoospermic patients. PMID- 16307339 TI - Can the complicated forgotten indwelling ureteric stents be lethal? AB - INTRODUCTION: The forgotten ureteral stents remain a urological dilemma and complications related to it can be lethal for the patient. The management of such stents require lithotripsy and endourological techniques. MATERIALS AND METHODS: We retrospectively reviewed the records of 19 cases of forgotten stents managed between 1998 and 2003. The mean patient age was 32 +/- l2 years, male to female ratio 17:2 and the mean duration of stents in urinary system was 24.2 months (7 months to l0 years). The stent were complicated in 14 patients and 5 patients had uncomplicated stents. The stents were severely calcified and encrusted in 6 patients, large stone formation seen at upper end of stent in 2 patients and at lower end of stents in 2 patients. The stents were spontaneously fragmented in 2 patients. The advanced renal failure secondary to hydroureteronephrosis because of severe encrustation and stone formation over the stent in solitary kidney was seen in 1 patient and 1 patient had upmigrated stent with infected hydronephrosis, but the duration of upmigration in this case was unknown. RESULTS: The stents were removed by retrograde approach in all 5 uncomplicated cases. In 6 patients of severely calcified and encrusted stents, the retrograde stent removal could be done in 4 patients while stent got broken in proximal ureter in 2 cases when they were being removed. In these 2 cases the stents were removed by percutaneous nephrostomy. The percutaneous nephrolithotomy and stent removal was done in 2 patients who had large stone at the upper end of stent in renal pelvis. In 2 patients who had large stone formation at lower end, stones were fragmented by mechanical lithotripsy in one and by laser lithotripsy in another case and stents removed by retrograde approach. Two cases of spontaneous fragmented stents were managed by retrograde endoscopic approach. The patient of advanced renal failure refused treatment and died. The patient of infective hydronephrosis with upmigrated stent died because of complications related to operative intervention. CONCLUSIONS: The management of complicated forgotten ureteral stents need judicious use of endourological techniques and lithotripsy. The stent related complication can be directly lethal for the patient or indirectly can cause death because of complications related to operative intervention. PMID- 16307340 TI - Atypical presentations of benign retroperitoneal schwannoma: report of three cases with review of literature. AB - We report 3 cases of benign retroperitoneal schwannoma, of whom one presented with flank pain with hematuria, one presented with headache and secondary hypertension and one presented with recurrent renal colicky pain. Two patients were treated by open surgical excision of the tumor with nephrectomy of the corresponding side because the tumors were densely adhered to the kidney and one case treated by laparoscopic resection of the mass only. The diagnosis of schwannoma was established postoperatively after histopathological examination and immunohistochemistry. All patients are doing well in follow-up. PMID- 16307341 TI - Spectrum of pulmonary infections in renal transplant recipients in the tropics: a single center study. AB - BACKGROUND: Pulmonary infections have been implicated as the most common cause of infection related mortality in renal transplant recipients. An appropriate empirical treatment of post transplant pulmonary infections requires knowledge of the spectrum of the microorganisms involved in causing these infections. Besides this knowledge, an aggressive diagnostic approach including the use of invasive tests is often essential to make an early diagnosis for instituting timely and appropriate therapy. We carried out a prospective cohort study to analyze the spectrum of pulmonary infections in these patients and study the utility of bronchoalveolar lavage (BAL) in the diagnosis of the same. METHODS: From September 2001 to December 2002, 428 patients were under follow up with the department. In all, 40 renal transplant recipients reported with 44 episodes of pulmonary infection during this study period. All patients underwent detailed and appropriate investigations including specific laboratory tests, sputum analysis, X-ray chest, CT and BAL. The spectrum of the causative organisms and the utility of BAL as compared to the other methods of diagnosis were studied and compared. RESULTS: Out of the 44 episodes of pulmonary infection evaluated, single causative organism could be found in only 24 (54.5%) episodes and multiple etiologies were found in 15 (34.1%) episodes. No definitive cause could be found in 5 episodes. Out of 57 organisms isolated in the 44 episodes, 20 (45.4 %) were bacteria, 16 (36.3 %) each were M. tuberculosis and fungus, 3 were CMV infection and 2 were nocardia. BAL gave a diagnostic yield of 75.8% (25 out of 33 cases). Nine of forty patients died (mortality rate 22.5%) of which 6 deaths could be attributed directly to pulmonary infection. Out of these 9 patients who died, cause of pulmonary infection was bacterial in 5, fungal in 2 and CMV disease in 1. In one patient, organism could not be isolated. CONCLUSIONS: Our study has shown that more than 1/3rd of pulmonary infections in renal transplant recipients can be attributed to multiple organisms. Bacterial infections were the commonest cause of post transplant pulmonary infection. Tuberculosis is common cause of pulmonary infection in these patients in our set up. Because of its high diagnostic yield, BAL should be considered in all patients with suspected pulmonary infections in the post transplant period. PMID- 16307342 TI - Fibrillary glomerulonephritis presenting as rapidly progressive renal failure in a young female: a case report. AB - Fibrillary Glomerulonephritis (FGN) is a rare clinical entity presenting in majority of patients with nephrotic range proteinuria, microscopic hematuria, impaired renal function and hypertension. The mean age of presentation is reported to be beyond 50 years with 50 patients developing end stage renal disease within a few years. A 28-year-old female presented to us with non nephrotic range proteinuria, rapidly progressive renal failure (RPRF), microscopic hematuria and mild hypertension. The patient had undergone a renal biopsy at a peripheral center, which was reported as membranoproliferative glomerulonephritis (MPGN) and was being treated with steroids without any benefit. The patient on re-biopsy at our center was diagnosed as FGN on electron microscopy (EM). The patient responded to intravenous pulse methylprednislone and oral cyclophosphamide and is off dialysis for 10 months now. This case highlights the varied clinical and histological presentations of FGN which makes the disease difficult to diagnose more so, in a country like ours where EM is available in only a few centers. An accurate diagnosis aided by high index of clinical suspicion and EM can help in the initiation of appropriate therapy, thereby improving the outcome in this disease which otherwise has a poor prognosis. PMID- 16307343 TI - Benefits of low dose immunoglobulin in the treatment of refractory CAPD peritonitis and longevity of technical survival on CAPD. AB - In this study we investigated the long term results of intraperitoneal immunoglobulin (Ig) treatment in continuous ambulatory peritoneal dialyses (CAPD) patients with refractory or relapsing peritonitis. Sixteen CAPD patients (4 female, 12 male) with a mean age of 53 +/- 11 years (40-80), with a mean CAPD duration of 46.2 +/- 4.8 months (17-75) were included in the study. The patients included had a diagnosis of either refractory or relapsing peritonitis unresponsive to appropriate antibiotic therapy. 0.5 g of Ig was added to every exchange bag qid as an adjunctive therapy to the culture based antibiotherapy for 7 days. Intraperitoneal Ig treatment was found to be successful in treating peritonitis in all but one patient. Interestingly, following Ig treatment, long term peritonitis rate decreased significantly compared to the period before treatment (before: 2.2 +/- 0.6 episodes/patient/year vs. after: 0.6 +/- 0.17 episodes/patient/year; P = 0.019). The mean CAPD duration after Ig treatment was 30.5 +/- 5.4 (4-64) months. Out of 16 patients, one patient who was unresponsive, had his catheter removed and was switched to hemodialysis, and four patients with preexisting ultrafiltration failure or inadequate dialysis problems were transferred to hemodialysis after successful treatment of their peritonitis, one patient was transplanted and 10 patients continued on CAPD. We conclude that low dose Ig treatment may be beneficial in the treatment of refractory or relapsing CAPD peritonitis possibly through restoring impaired host defense within peritoneal cavity. This therapy, by preventing further peritonitis attacks, may prolong survival on CAPD. PMID- 16307344 TI - Hydrothorax due to ventriculopleural shunting in a child with spina bifida on chronic dialysis: third ventriculostomy as an alternative of cerebrospinal diversion. AB - The purpose of this paper is to describe the risks of ventriculopleural shunt in patients with spina bifida and end-stage-renal-diseases (ESRD), and to describe endoscopic third ventriculostomy as an alternative for the combination of cerebrospinal shunt and dialysis modality. We report a 16-year-old boy with spina bifida on chronic dialysis with a massive unilateral hydrothorax and respiratory distress complicating a ventriculopleural (VPL) shunt. Two thoracocenteses were performed, draining 3200 ml of a clear fluid. The VPL shunt was removed and revised successfully to a third ventriculostomy (TVE). Peritoneal dialysis (PD) was the initial dialysis modality. After 12 months on PD, the patient was transferred to hemodialysis (HD) because of refractory peritonitis. Hydrothorax developed while the patient was on PD, reaching its maximum 2 months after the transference to HD. To our knowledge there has been no other report of ventriculopleural (VPL) shunt failure, and endoscopic TVE, as a cerebrospinal fluid (CSF) diversion alternative in patients on chronic dialysis. PMID- 16307345 TI - Volume control in diabetic and nondiabetic peritoneal dialysis patients. AB - Diabetic patients especially the elderly have higher mortality on peritoneal dialysis treatment as compared to nondiabetic patients. As fluid overload is an important contributor for the high dropout rate in peritoneal dialysis therapy, the present study tried to compare the fluid status in diabetic and non-diabetic peritoneal dialysis patients and to investigate the effect of salt and water restriction on fluid status in diabetic peritoneal dialysis patients. METHODS: A cross sectional study was performed in 58 peritoneal dialysis patients who had been treated for at least 3 months. Among them, 20 patients were diabetics and the others were nondiabetics. We reviewed their dialysis prescription, peritoneal ultrafiltration, urine volume and fluid status. Then the 20 diabetic patients were advised to restrict their salt and water intake and were followed for 3 months. RESULTS: During the cross sectional study there were no significant differences in age, gender, height and urine volume between the diabetic and nondiabetic patients. However, body weight, normalized extracellular water (nECW), dialysis dose, dialysate glucose load, peritoneal ultrafiltration, total fluid removal and the prevalence and extent of edema were all significantly higher in diabetics as compared to nondiabetics. After restricting salt and water intake, body weight, nECW, edema, fasting blood glucose, dialysate dose, dialysate glucose load, peritoneal ultrafiltration and total fluid removal were all significantly decreased, whereas the urine volume did not change significantly. CONCLUSIONS: The present study suggests that diabetic patients were more fluid overloaded as compared to nondiabetics despite use of more hypertonic glucose solutions and, as a consequence, higher peritoneal ultrafiltration and higher total fluid removal; this indicates that our diabetic patients must have had significantly higher salt and fluid intakes. It also suggests that restricting salt and water intake can effectively treat fluid overload in diabetic peritoneal dialysis patients. Dietary salt and fluid restriction may help reduce the use of hypertonic glucose solution and thus facilitate the blood glucose control in diabetic patients undergoing peritoneal dialysis. PMID- 16307346 TI - An atypical case of POEMS syndrome with IgG kappa M protein and end stage renal failure. AB - POEMS syndrome is a rare plasma cell dyscrasia which is characterized by small amounts of monoclonal protein, and a multisystem complex manifested by various combinations of polyneuropathy, organomegaly, endocrinopathy and skin changes. Here, we presented an atypical case of POEMS syndrome with IgG kappa monoclonal protein, chronic demyelinating polyneuropathy, hepatosplenomegaly, hypothyroidism, gynecomastia and severe renal impairment. The finding of IgG kappa type of monoclonal protein in our patient was interesting because the majority of cases were reported to have lambda light chain. Also, the absence of typical skin and bone lesions were atypical. Though speculative, these atypical features may account for the unusual presentation of this case. Our patient rapidly progressed to end-stage renal failure and died of cachexia. Renal involvement in POEMS syndrome is rare but may show substantial clinical and pathological variations. Proteinuria, hematuria, renal dysfunction and renal failure requiring hemodialysis can be seen. The pathogenesis of renal dysfunction is unclear. As a conclusion, POEMS syndrome may present with diverse clinicopathologic manifestations. In this syndrome, renal involvement may lead to end stage renal failure and the course may be fatal due to severe polyneuropathy and wasting. PMID- 16307347 TI - Protective effect of oral L-arginine supplementation on cyclosporine induced nephropathy in rats. AB - BACKGROUND: One of the major adverse effects of long term cyclosporine A (CyA) administration is chronic nephrotoxicity. Several studies have suggested that alterations of the L-arginine (L-Arg) nitric oxide (NO) pathway may be involved in the pathogenesis of CyA-induced kidney damage. AIM: We postulated that in vivo activation of L-Arg-NO pathway might have a beneficial effect on CyA-induced renal damage. Conditions of chronic NO enhancement was established with L-Arg supplementation and chronic NO blockade with N-nitro-L-Arg methyl ester (L-NAME). We tested the hypothesis that, if CyA administration alters intrarenal NO synthesis, then exogenous L-Arg supplementation could limit renal injury, on the contrary, L-NAME, a potent competitive inhibitor of NO synthesis, could enhance CyA nephrotoxicity. Harmful effect of NO blockade indirectly supports the beneficial effect of NO in a model of CyA nephrotoxicity. METHODS: Rats were administered vehicle (VH), CyA (7.5 mg/kg/day), CyA + L-Arg (2g/kg/day), CyA + L NAME (5 mg/100 ml/day), CyA + L-Arg + L-NAME, VH + L-Arg, VH + L-NAME and were sacrificed at the end of the experiment. Body weight, serum creatinine, blood urea nitrogen (BUN) and NO levels were determined. Tubular injury and interstitial fibrosis were evaluated semiquantitatively using scoring systems on paraffin sections stained with hematoxylin/eosin (H/E), Masson's trichromic and periodic acid-Schiff (PAS). RESULTS: The CyA group developed marked renal injury, characterized by a significant increase in serum creatinine and BUN, and histopathological alterations including tubular dilatation, vacuolization, necrosis, interstitial cell infiltration and tubulointerstitial fibrosis. CyA reduced serum NO level. L-Arg treatment significantly enhanced NO biosynthesis and protected animals from CyA-induced kidney damage. In contrast L-NAME strikingly reduced serum NO level, and worsened biochemical and histopathological alterations. CONCLUSION: Chronic CyA nephrotoxicity can be aggravated by NO blockade and ameliorated by NO enhancement suggesting that L-Arg supplementation may be protective in CyA nephrotoxicity. PMID- 16307348 TI - Quality of life in Turkish haemodialysis patients. AB - The aim of the study was to assess quality of life (QOL) in Turkish haemodialysis patients and to identify related socio-demographic and clinical variables. To measure QOL 100 patients completed SF-36 during regularly scheduled haemodialysis. We found that patients' QOL was substantially impaired. Age was negatively related to physical components of QOL. Duration of haemodialysis was negatively correlated with most of QOL dimensions. EPO treatment, education on disease and haemodialysis, and compliance to prescribed diet had great positive effects on QOL. Among the factors we have found to be related with the QOL in this study, probably the most important were education on disease and haemodialysis, and compliance to prescribed diet, in which we have influence, as nurses, on improving the QOL in patients receiving haemodialysis. The priority of renal nursing services should therefore be addressed to provide support in these areas. PMID- 16307349 TI - Acute postinfectious crescentic glomerulonephritis: clinicopathologic presentation and risk factors. AB - BACKGROUND: Glomerular crescent formation is a feature of the most severe forms of human glomerulonephritis. The postinfectious form of rapidly progressive glomerulonephritis with crescents is a form of immune complex glomerulonephritis which seem to have a better prognosis. A relatively poorer prognosis for crescentic postinfectious glomerulonephritis in South Africa has been reported. In the present study, we have tried to determine the mode of presentation, and the prognostic factors for renal and patient outcome for cases with postinfectious crescentic glomerulonephritis (CGN). METHODS: Between 1990 and 2000 a total number of 128 patients with CGN were managed at our center, among them 23 cases were diagnosed as postinfectious CGN. They were followed-up for a mean period of 40.1 +/- 28.9 months. Among them 12 were males and 11 were females. The median age was 12.35 years (range 4-55 years). The median serum creatinine at presentation was 7.24 mg/dl (range 1.3-14.5 mg/dl). We studied the clinical, laboratory and histopathological data .of our cases and their impact on the renal and patient outcome. RESULTS: By univariate study the risk factors for renal dysfunction were the age, hypertension, and nephrotic range proteinuria during the follow-up period. By multivariate analysis only the, hypertension, and presence of nephrotic range proteinuria during the follow-up period were the significant risk factors. The risk factors that significantly affected patient mortality were hypertension and serum creatinine at last follow-up. CONCLUSION: postinfectious CGN is a severe form of glomerulonephritis that usually presents with rapidly progressive renal failure. The persistence of hypertension and nephrotic range proteinuria during the follow-up are major bad prognostic predictors for renal dysfunction. PMID- 16307350 TI - Dissociation between the correlation of peritoneal and urine Kt/V with sodium and fluid removal: a possible explanation of their difference on patient survival. AB - BACKGROUND: It has been shown that residual renal function but not peritoneal clearance predicted patients' survival in peritoneal dialysis therapy. In the present study, we tried to explore the potential causes resulting in the difference between residual renal function and peritoneal dialysis in continuous ambulatory peritoneal dialysis (CAPD) patients. METHODS: A cross sectional study was performed during July and August 2003 to evaluate the dialysis adequacy in CAPD patients who were clinically stable and had daily urinary volume more than 100 ml. RESULTS: A total of 45 patients (male 27 and female 18) with an average ( +/- SD) age of 61.76 +/- 13.27 years were included in this study. The daily urinary volume and dialysate ultrafiltration volume were 570.33 +/- 395.47 ml and 726.09 +/- 454.01 ml, respectively. Peritoneal urea clearance (Kt/V) correlated significantly with the drained daily dialysate volume (r = 0.362, P < 0.01), but not with peritoneal net fluid removal (ultrafiltration) (r = 0.232, P > 0.05) and sodium removal (r = 0.139, P > 0.05). On the other hand, there were strong positive correlations between residual renal Kt/V and daily urine volume (r = 0.802, P < 0.001), as well as between residual renal Kt/V and urinary sodium removal (r = 0.670, P < 0.001). CONCLUSIONS: High residual renal Kt/Vurea represents both higher solute clearance and higher sodium and fluid removal, but higher peritoneal Kt/Vurea is not necessarily associated with better sodium and fluid removal. This dissociation might explain the differences on the survival of patients and peritoneal clearances. PMID- 16307351 TI - Does mycophenolate mofetil decrease the recurrent acute rejection in renal transplant recipients. AB - PURPOSE: Mycophenolate mofetil (MMF) has emerged as a valuable immunosuppression in renal transplant patients. However, it is expensive and cannot be used routinely in our population. MATERIAL AND METHODS: In a retrospective study, 60 renal transplant patients on MMF based triple immunosuppression were analysed. The indication for MMF was as rescue therapy after treatment of acute vascular rejection (Banff type-4, grade IIA, IIB and III) in all patients. However, 20 such patients also had associated chronic liver diseases. The patients were given 1.5-2.0 g MMF in two divided doses at least for 6 months, depending upon the tolerability, adverse effects and affordability, and followed-up at least for 1 year. The control group consisted of 60 cases of acute vascular rejection (Banff type-4, grade IIA, IIB and III) who were placed on cyclosporine, azathioprine and steroid based maintenance immunosuppressive regimen in same time frame. RESULTS: The incidence recurrent acute rejections in MMF group was 18% and 42% in control group (P < 0.005). The serum transaminases in all patients of the liver diseases became normal in 3-6 months. The incidence of opportunistic infections in MMF and control group were 22% and 11% respectively (P < 0.05). The MMF based regimen was two times more expensive. The 1 year patient and graft survivals between two groups were not statistically significantly different. CONCLUSION: The MMF based regimen significantly decreases the recurrent acute rejections. However, it is expensive and cannot be used routinely in all patients in Indian scenario. PMID- 16307352 TI - Anthranilic acid-uraemic toxin damaged red cell's membrane. AB - Normocytic normochromic anaemia is a common syndrome present in patients with chronic renal insufficiency (CRI). Simultaneously in these patients the increase in L-tryptophan (TRP) degradation via kynurenine pathway is observed. On the basis of these observations we tried to examine whether one of the TRP metabolites, anthranilic acid (AA), shows interaction with membranes of erythrocytes and because of that it may contribute to anaemia development. In patients with CRI we have observed changes characteristic for normocytic normochromic anaemia, such as the decrease in erythrocyte count, haemoglobin concentration, haematocrit and the decrease in erythrocyte osmotic resistance as well as the increase in AA concentration in plasma in comparison to healthy subjects. We have also noticed the existence of a positive correlation between anthranilic acid concentration and creatinine and urea concentrations and also negative relationships between anthranilic acid concentration and haematological parameters. Moreover, incubation of healthy erythrocytes with 10 and 100 microM AA caused haemolysis curve movement to the right, which shows decrease in osmotic resistance. In conclusion, the increase in plasma AA concentration might be one of many factors, which damage erythrocyte membrane, and thereby contributes to anaemia development in patients with CRI. PMID- 16307353 TI - Diabetic muscle infarction: an unusual cause of muscle pain in a diabetic patient on hemodialysis. AB - Diabetic muscle infarction (DMI) is a rare, painful and potentially serious complication in patients with poorly controlled diabetes mellitus and frequently misdiagnosed clinically as abscess, neoplasm, or myositis. A 36-year-old diabetic woman referred to our clinic with severe pain in the left antero-medial thigh. She had a 15-year history of Type 2 diabetes mellitus (DM). She was complicated by diabetic nephropathy and requiring hemodialysis. She had first noticed pain and swelling in her left thigh after a minimal trauma for 2 days prior to presentation. Clinical and laboratory evaluation, and muscle biopsy revealed the diagnosis of muscle infarctions. She did no respond to the conservative therapy. Pain and swelling in her thigh worsened progressively. She underwent surgical debridment and then, her clinical status improved. We describe the characteristic clinical and pathologic findings and the course of the illness with emphasis on the importance of recognition of the syndrome so that unnecessary investigation and overzealous therapy can be avoided. PMID- 16307354 TI - Co-administration of ketoconazole and tacrolimus therapy: a transplanted rat model. AB - BACKGROUND/AIM: The aim of this work is to study the safety and the effect of addition of ketoconazole to experimental kidney transplanted rat treated with tacrolimus and predicts the percentage of tacrolimus dose reduction. MATERIAL AND METHODS: The material of this work included 60 male Sprague Dawely rats subjected to renal allotransplantation. They were equally divided into five groups: Group I: served as control group, Group II: received FK506 3.2 mg/kg/bw, Group III: received FK506 2 mg/kg/bw, Group IV: received FK506 1 mg/kg/bw, Group V: received FK506 1 mg/kg/bw plus Ketoconazole 20 mg/kg/day. FK506 trough level and laboratory investigations were determined at 0, 3, 7, 10, 14, and 27 days post transplantation. RESULTS: In all groups loss of body weight was observed at day 27 after treatment compared to that before transplantation. Serum creatinine significantly increased at day 27 compared to the basal level in groups treated with 1.0 and 3.2 mg FK506 (1.80 +/- 0.50 versus 0.39 +/- 0.06 P = 0.001) and (1.03 +/- 0.26 versus 0.50 +/- 0.07 P = 0.001) respectively, while for 2.0 mg or 1.0 mg plus keto groups, no significant differences in serum creatinine levels over time (0.56 +/- 0.22 versus 0.44 +/- 0.10 P = 0.106) and (0.55 +/- 0.30 versus 0.42 +/- 0.08 P=0.160) were observed. CONCLUSION: Concomitant administration of Ketoconazole and FK506 in transplanted rat model is safe and results in increase of blood trough level concentration of FK506 with 50% reduction of its dose. PMID- 16307355 TI - Live and deceased donor kidney transplantation in patients aged 75 years and older in the United States. AB - OBJECTIVE: To examine the outcomes of geriatric ESRD patients selected for kidney transplantation. DESIGN: Data were extracted from the USRDS Standard Analysis Files (SAF). All persons ages 75 and over who received a kidney transplant from 1994 to 2000 were compared with those remaining on dialysis or on a transplant waiting list. Data on mortality or removal from the waiting list were obtained from the United Network for Organ Sharing (UNOS). The main outcome measure was patient and kidney transplant survival. RESULTS: Superior five year survival after kidney transplantation was attained by the geriatric cohort given a live donor transplant (59.9%), compared with recipients of deceased donor kidneys (40.3%), dialysis patients waiting for transplant (29.7%), and those who were not selected for kidney transplantation and remained on dialysis (12.5%). The likelihood of being removed from the waiting list for any reason was higher in this group (over 75) (30.3%) than in the 66-75 age group (26.8%). Their average annual mortality rate on the waiting list was 7.9, compared to 6.6% for those 66 75. CONCLUSION: Even after the age of 75 years, kidney transplantation provides substantial life prolongation and excellent graft survival. PMID- 16307356 TI - Prevalence of falls among seniors maintained on hemodialysis. AB - BACKGROUND: Increasing numbers of seniors are starting renal replacement therapy. Many may experience a loss of functional independence including mobility impairment. Seniors on dialysis commonly have multiple comorbidities that are associated with an increased risk for falls in the general population. Falls lead to serious injury, loss of independence, hospitalization and institutionalization. Despite such morbidity, little attention has been directed toward this marker of frailty in the dialysis population. In this brief preliminary report, we set out to determine the prevalence of falls among seniors on dialysis. METHODS: A cross-sectional interview based survey was used to determine one year fall prevalence in patients aged 65 years and older receiving in center hemodialysis therapy. RESULTS: A total of 135 ambulatory seniors were interviewed. Thirty-seven (27%) people reported having fallen in the past 12 months. An additional 21(16%) reported a fall prior to the past year. Four percent recalled a fall associated fracture or head injury requiring treatment. CONCLUSIONS: Falls are very common amongst seniors on hemodialysis. Further prospective study of the incidence and risk factors for falls in this population is needed to allow targeted interventions. PMID- 16307357 TI - Renal physiology in the oldest old: the Sphinx remakes her question. PMID- 16307358 TI - Prevention of nephropathy in patients with type 2 diabetes mellitus. AB - The rising incidence of type 2 diabetes mellitus and of its complications will make it the most important health care challenge in the first quarter of the 21st Century. Diabetic nephropathy left unchecked will overwhelm the renal resources. Simple methods (proper diet and exercise, prevention of obesity) are successful in preventing type 2 diabetes in the great majority of the persons at risk. In patients with established type 2 diabetes, nephropathy can be prevented or greatly delayed by strict metabolic control, strict control of blood pressure using angiotensin-converting enzyme inhibitors and angiotensin receptor blockers as the first line of drugs, tight control of serum lipids using statins as indicated, low protein diet, avoidance of smoking and other nephrotoxic influences, prevention of abnormalities in calcium/phosphorus metabolism, and prevention of renal anemia by the early use of erythropoietin. Current research offers the promise of definitive prevention of both type 2 diabetes and diabetic nephropathy. PMID- 16307359 TI - Clinical Pharmacology and Physiology Conference: digoxin toxicity in the elderly. PMID- 16307360 TI - A clinical, physiology and pharmacology evaluation of orthostatic hypotension in the elderly. AB - Orthostatic hypotension is very common in the elderly. It increases morbidity and is an independant predictor of all cause mortality. It is defined as a fall in systolic blood pressure greater than 20 mm Hg or a fall in diastolic blood pressure greater than 10 mm Hg within 3 minutes of standing. Symptoms include light headedness, weakness, blurred vision, fatigue and lethargy and falls. Most patients have orthostatic hypotension due to non neurogenic causes. Drugs like antihypertensives and tricyclic antidepressants are very common causes of orthostatic hypotension. Diagnosis is based on the history and a thorough clinical examination. Based on the history and physical examination, further testing of the heart, kidneys and autonomic nervous system may be required in selected patients. Non pharmacological methods like slow position change, increased fluid and sodium intake, compression stockings and elevation of head of the bed are the key to management of orthostatic hypotension. After these methods, pharmacological treatment with fludrocortisone and midodrine should be tried. Other drugs like desmopresin acetate, xamoterol, erythropoetin and ocreotide can be used as second line agents in selected patients. PMID- 16307361 TI - The genomic organization of retrotransposons in Brassica oleracea. AB - We have investigated the copy numbers and genomic organization of five representative reverse transcriptase domains from retrotransposons in Brassica oleracea. Two non-homologous Pseudoviridae (Ty1/copia-like) elements, two Metaviridae (Ty3/gypsy-like) elements (one related to the Athila family) and one Retroposinae (LINE) element were hybridized to a gridded BAC library, "BoB". The results indicated that the individual LTR retrotransposons (copia and gypsy-like) were represented by between 90 and 320 copies in the haploid genome, with only evidence of a single location for the LINE. Sequence analysis of the same elements against genome survey sequence gave estimates of between 60 and 570, but no LINE was found. There was minimal evidence for clustering between any of these retroelements: only half the randomly expected number of BACs hybridized to both LTR-retrotransposon families. Fluorescent in situ hybridization showed that each of the retroelements had a characteristic genomic distribution. Our results suggest there are preferential sites and perhaps control mechanisms for the insertion or excision of different retrotransposon groups. PMID- 16307362 TI - An annotation update via cDNA sequence analysis and comprehensive profiling of developmental, hormonal or environmental responsiveness of the Arabidopsis AP2/EREBP transcription factor gene family. AB - AP2/EREBP transcription factors (TFs) play functionally important roles in plant growth and development, especially in hormonal regulation and in response to environmental stress. Here we reported verification and correction of annotation through an exhaustive cDNA cloning and sequence analysis performed on 145 of 147 gene family members. A RACE analysis performed on genes with potential in-frame up-stream ATG codon resulted in identification of At2g28520 as an authentic AP2/EREBP member and corrected ORF annotations for three other members. A further phylogenetic analysis of this updated and likely complete family divided it into three major subfamilies. The expression patterns of the AP2/EREBP family members among the 11 organ or tissue types were examined using an oligo microarray and their hormonal and environmental responsiveness were further characterized using cDNA custom macroarrays. These detailed expression profile results provide strong support for a role for AP2/EREBP family members in development and in response to environmental stimuli, and a foundation for future functional analysis of this gene family. PMID- 16307363 TI - Endosperm-specific co-expression of recombinant soybean ferritin and Aspergillus phytase in maize results in significant increases in the levels of bioavailable iron. AB - We have generated transgenic maize plants expressing Aspergillus phytase either alone or in combination with the iron-binding protein ferritin. Our aim was to produce grains with increased amounts of bioavailable iron in the endosperm. Maize seeds expressing recombinant phytase showed enzymatic activities of up to 3 IU per gram of seed. In flour paste prepared from these seeds, up to 95% of the endogenous phytic acid was degraded, with a concomitant increase in the amount of available phosphate. In seeds expressing ferritin in addition to phytase, the total iron content was significantly increased. To evaluate the impact of the recombinant proteins on iron absorption in the human gut, we used an in vitro digestion/Caco-2 cell model. We found that phytase in the maize seeds was associated with increased cellular iron uptake, and that the rate of iron uptake correlated with the level of phytase expression regardless of the total iron content of the seeds. We also investigated iron bioavailability under more complex meal conditions by adding ascorbic acid, which promotes iron uptake, to all samples. This resulted in a further increase in iron absorption, but the effects of phytase and ascorbic acid were not additive. We conclude that the expression of recombinant ferritin and phytase could help to increase iron availability and enhance the absorption of iron, particularly in cereal-based diets that lack other nutritional components. PMID- 16307364 TI - The maize An2 gene is induced by Fusarium attack and encodes an ent-copalyl diphosphate synthase. AB - Using the technique of differential display, a maize transcript was identified whose silk tissue expression is induced in the presence of the ear rot pathogen Fusarium graminearum. The 3445 nt transcript includes a 727 nt 5' untranslated leader with the potential for extensive secondary structure and represents the maize gene An2. An2 encodes a copalyl diphosphate synthase (CPS)-like protein with 60% amino acid sequence identity with the maize An1 gene product involved in gibberellin (GA) biosynthesis. Recombinant expression and functional analysis demonstrated that both AN1 and AN2 are ent-copalyl diphosphate (ent-CPP) synthases (ent-CPS). Notably, the presence of an additional ent-CPS gene is consistent with previous reports that maize GA biosynthesis can proceed in the absence of An1. In addition, northern blot analysis showed that An2 transcript levels were strongly up-regulated by Fusarium attack, with an increase in silk, husk and ear tip tissues as early as 6 h after inoculation of silk channels with spore suspensions of various Fusarium sp. Gene expression of a third maize CPS like gene, Cpsl1, is not affected by Fusarium infection. The Fusarium-inducible nature of An2 is also consistent with a previous report that cell-free extracts from maize seedlings produce ent-CPP derived diterpenes in response to Fusarium infection. However, it is not known whether An2 is involved in defense-related secondary metabolism in addition to GA synthesis. PMID- 16307365 TI - Comparative analysis of expressed sequence tags of conifers and angiosperms reveals sequences specifically conserved in conifers. AB - To identify and characterize lineage-specific genes of conifers, two sets of ESTs (with 12791 and 5902 ESTs, representing 5373 and 3018 gene transcripts, respectively) were generated from the Cupressaceae species Cryptomeria japonica and Chamaecyparis obtusa. These transcripts were compared with non-redundant sets of genes generated from Pinaceae species, other gymnosperms and angiosperms. About 6% of tentative unique genes (Unigenes) of C. japonica and C. obtusa had homologs in other conifers but not angiosperms, and about 70% had apparent homologs in angiosperms. The calculated GC contents of orthologous genes showed that GC contents of coniferous genes are likely to be lower than those of angiosperms. Comparisons of the numbers of homologous genes in each species suggest that copy numbers of genes may be correlated between diverse seed plants. This correlation suggests that the multiplicity of such genes may have arisen before the divergence of gymnosperms and angiosperms. PMID- 16307366 TI - Dynamic histone acetylation of late embryonic genes during seed germination. AB - Histone acetylation is involved in the regulation of gene expression in plants and eukaryotes. Histone deacetylases (HDACs) are enzymes that catalyze the removal of acetyl groups from histones, which is associated with the repression of gene expression. To study the role of histone acetylation in the regulation of gene expression during seed germination, trichostatin A (TSA), a specific inhibitor of histone deacetylase, was used to treat imbibing Arabidopsis thaliana seeds. GeneChip arrays were used to show that TSA induces up-regulation of 45 genes and down-regulation of 27 genes during seed germination. Eight TSA-up regulated genes were selected for further analysis - RAB18, RD29B, ATEM1, HSP70 and four late embryogenesis abundant protein genes (LEA). A gene expression time course shows that these eight genes are expressed at high levels in the dry seed and repressed upon seed imbibition at an exponential rate. In the presence of TSA, the onset of repression of the eight genes is not affected but the final level of repressed expression is elevated. Chromatin immunoprecipitation and HDAC assays show that there is a transient histone deacetylation event during seed germination at 1 day after imbibition, which serves as a key developmental signal that affects the repression of the eight genes. PMID- 16307367 TI - Identification of NPR1-dependent and independent genes early induced by salicylic acid treatment in Arabidopsis. AB - Salicylic acid (SA) plays a crucial role in stress resistance in plants by modifying the expression of a battery of genes. In this paper, we report the identification of a group of early SA-regulated genes of Arabidopsis (activated between 0.5-2.5 h), using the cDNA-amplified fragment length polymorphism technique (cDNA-AFLP). Using 128 different primer combinations, we identified several genes based on their differential expression during SA treatment. Among these, we identified 12 genes up-regulated by SA whose patterns of induction were confirmed by Northern analysis. The identified genes can be grouped into two functional groups: Group 1: genes involved in cell protection (i.e. glycosyltransferases, glutathion S-transferases), and Group 2: genes involved in signal transduction (protein kinases and transcription factors). We also evaluated NPR1 requirement for the induction of the 12 up-regulated genes, and found that only those belonging to Group 2 require this co-activator for their expression. In silico analysis of the promoter sequences of the up-regulated genes, allowed us to identify putative cis-elements over-represented in these genes. Interestingly, as-1-like elements, previously characterized as SA responsive elements, were specifically over-represented in Group 1 genes. The identification of early SA-regulated genes is an important step towards understanding the complex role of this hormone in plant stress resistance. PMID- 16307368 TI - Tissue-specific expression and drought responsiveness of cell-wall invertase genes of rice at flowering. AB - Drought stress near heading reduces grain yield in rice cultivars by inhibiting processes such as anther dehiscence and panicle exsertion. Because cell-wall invertases play an important role in carbon allocation to developing organs, we examined the tissue-specific expression and drought sensitivity of the corresponding genes (OsCIN1-9) at heading in the widely grown cultivar IR64. OsCIN1-5,8 were expressed to varying degrees in flag leaf, panicle, anthers and peduncle at 1 day before heading (1 DBH). When water was withheld for 2 days starting 3 DBH, anthesis and peduncle elongation were halted. At the same time, transcript levels for OsCIN1-5,8 genes were all markedly down-regulated in anthers and/or peduncles but were not affected in flag leaves. Re-watering allowed anthesis and peduncle elongation to proceed and restored expression of OsCIN1-5,8. We conclude that cell-wall invertase genes, as a class, respond rapidly to water deficit in anthers and peduncles and through a reduction in sink strength help to coordinate a delay in anthesis and heading. By contrast, vacuolar invertase OsVIN2 was up-regulated by drought stress in flag leaves, panicles, anthers and peduncles. Although OsCIN1-3,5,8 were active in the peduncle, only OsCIN2 was expressed strongly and preferentially at the base, where cell division and cell elongation occur. OsCIN2 was expressed principally in the primary and secondary vascular systems, consistent with a role in diverting sucrose from the phloem to the dividing and expanding cells of the peduncle, whereas the less abundant OsCIN1,3,5,8 transcripts were found principally in parenchyma cells. The OsCIN2 transcript levels in the base were highest at 1 DBH, when rapid peduncle elongation began. Drought stress halted peduncle elongation and reduced OsCIN2 transcript level to 8% of the control level. On re-watering, peduncle elongation was restored and OsCIN2 transcript level recovered to 24% of the control. The abscisic acid (ABA) level of peduncles increased 7-fold on drought stress and returned to the control level on re watering. Detached peduncles floated on water elongated little and lost all OsCIN2 transcripts, but on 50-100 microM GA3 they elongated rapidly and maintained high OsCIN2 transcript levels. ABA antagonized both peduncle elongation and maintenance of OsCIN2 transcript levels. We conclude that this antagonism is a potential intervention point for breeding strategies directed at enhancing panicle exsertion during or after drought stress at heading. PMID- 16307369 TI - Laser capture microdissection (LCM) and expression analyses of Glycine max (soybean) syncytium containing root regions formed by the plant pathogen Heterodera glycines (soybean cyst nematode). AB - Roots of soybean, Glycine max cv. Kent L. Merr., plants susceptible to the soybean cyst nematode (SCN), Heterodera glycines Ichinohe, were inoculated and allowed to develop feeding sites (syncytia) for 8 days. Root samples enriched in syncytial cells were collected using laser capture microdissection (LCM). RNA was extracted and used to make a cDNA library and expressed sequence tags (ESTs) were produced and used for a Gene Ontology (GO) analysis. RT-PCR results indicated enhanced expression of an aquaporin (GmPIP2,2), alpha-tubulin (GmTubA1), beta tubulin (GmTubB4) and several other genes in syncytium-enriched samples as compared to samples extracted from whole roots. While RT-PCR data showed increased transcript levels of GmPIP2,2 from LCM tissue enriched in syncytial cells, in situ hybridization showed prominent GmPIP2,2 hybridization to RNA in the parenchymal cells tightly juxtaposed to the syncytium. Immunolocalization indicated stronger alpha-tubulin signal within the syncytium as compared to surrounding tissue. However, alpha-tubulin labeling appeared diffuse or clumped. Thus, LCM allowed for the isolation of tissue enriched for syncytial cells, providing material suitable for a variety of molecular analyses. PMID- 16307370 TI - In vitro and in planta interaction evidence between Nicotiana tabacum thaumatin like protein 1 (TLP1) and cucumber mosaic virus proteins. AB - Using a yeast two-hybrid system, we identified a plant cellular factor that interacts with the proteins of the Cucumber mosaic virus (CMV). Initially 14 candidate genes were isolated from Nicotiana tabacum, using a full-length CMV 1a gene as bait. Among the candidate genes, two were encoding thaumatin-like proteins (TLP), and were designated as Nicotiana tabacum thaumatin-like protein 1 (NtTLP1). Consistent with this observation, recombinant GST-NtTLP1 protein, which was expressed and purified in E. coli, bound tightly to CMV 1a in vitro. In planta interaction was also verified via co-immunoprecipitation. Additionally, NtTLP1 specifically interacted with the CMV movement-related proteins, movement protein and coat protein, in yeast. Real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis showed that the expression of NtTLP1 increased as the result of CMV inoculation. PMID- 16307374 TI - Soy isoflavones reduce heat shock proteins in experimental atherosclerosis. AB - BACKGROUND: Soy isoflavones may affect several biochemical pathways like the synthesis of nitric oxide (*NO) and heat shock proteins (HSP) that are important factors for atherosclerosis development. THE AIM OF THE STUDY: The purpose of this study was to investigate the influence of soy isoflavones on the production of *NO and HSP60, HSP70 and HSC70 in experimental atherosclerosis. METHODS: One group of rabbits (New Zealand) was fed an atherogenic diet containing 27 % casein (CAS) and another group was fed the same diet supplemented with soy isoflavones (5 mg/kg/day) (ISO). Blood samples were obtained monthly and after six months of feeding, the rabbits were sacrificed and the aortas were removed. RESULTS: The ISO group showed a significant reduction of cholesterol in LDL (36.2 %) and in aorta (36 %), as well as, an increase of HDL-cholesterol (2.1 times) in relation to the CAS group. The concentration of *NO metabolites (NOx) in blood plasma and the levels of reactive antibodies to HSC70 in blood plasma and to HSC70 and HSP70 in aortic tissue were significantly decreased in the ISO group. Isoflavones promoted a reduction of content of HSP60, HSP70 and HSC70 in aortic arch analyzed by immunohistochemistry. The isoflavone supplementation promoted a reduction of cholesterol content in aorta (62.2 %) (p < 0.05). CONCLUSIONS: Soy isoflavones reduced hypercholesterolemia, the production of HSP60, HSC70 and HSP70 and reactive antibodies to HSC70 in serum and to HSC70 and HSP70 in aorta, as well as, the cholesterol content in atherosclerotic lesions in rabbits fed a casein based atherogenic diet. PMID- 16307375 TI - Domain formation in DODAB-cholesterol mixed systems monitored via Nile Red anisotropy. AB - The effect of the cholesterol (Ch) on liposomes composed of the cationic lipid dioctadecyldimethylammonium bromide (DODAB) was assessed by studying both the steady-state and time-resolved fluorescence anisotropy of the dye Nile Red. The information obtained combined with analysis of the steady-state emission and fluorescence lifetime of Nile Red (NR) for different cholesterol concentrations (5-50%) elucidated the presence of "condensed complexes" and cholesterol-rich domains in these mixed systems. The steady-state fluorescence spectra were decomposed into the sum of two lognormal emissions, emanating from two different states, and the effect of temperature on the anisotropy decay of Nile Red for different cholesterol concentrations was observed. At room temperature, the time resolved anisotropy decays are indicative of NR being relatively immobile (manifest by a high r (infinity) value). At higher temperature, rotational times ca. 1 ns were obtained throughout and a trend in increasing hindrance was seen with increase of Ch content. PMID- 16307377 TI - Chemical genetics: Drug screens in Zebrafish. AB - High throughput chemical genetic screens for compounds with specific biological activity in a whole organism are feasible using zebrafish embryos. At least two medium to large scale drug screens have been carried out to date, leading to the identification of compounds that disturb zebrafish development. Chemical genetics using zebrafish embryos may become an important step in the discovery of drugs and their targets. PMID- 16307378 TI - Uncover genetic interactions in Caenorhabditis elegans by RNA interference. AB - RNA-mediated interference (RNAi) has emerged recently as one of the most powerful functional genomics tools. RNAi has been particularly effective in the nematode worm C. elegans where RNAi has been used to analyse the loss-of-function phenotypes of almost all predicted genes. In this review, we illustrate how RNAi has been used to analyse gene function in C. elegans as well as pointing to some future directions for using RNAi to examine genetic interactions in a systematic manner. PMID- 16307379 TI - Two sides of the same coin: Wnt signaling in neurodegeneration and neuro oncology. AB - Wnts function through the activation of at least three intracellular signal transduction pathways, of which the canonical beta-catenin mediated pathway is the best understood. Aberrant canonical Wnt signaling has been involved in both neurodegeneration and cancer. An impairment of Wnt signals appears to be associated with aspects of neurodegenerative pathologies while overactivation of Wnt signaling is a common theme in several types of human tumors. Therefore, although therapeutic approaches aimed at modulating Wnt signaling in neurodegenerative and hyperproliferative diseases might impinge on the same molecular mechanisms, different pharmacological outcomes are required. Here we review recent developments on the understanding of the role of Wnt signaling in Alzheimer's disease and CNS tumors, and identify possible avenues for therapeutic intervention within a complex and multi-faceted signaling pathway. PMID- 16307380 TI - Sphingolipids: modulators of HIV-1 infection and pathogenesis. AB - HIV-1 infects host cells by sequential interactions of its fusion protein (gp120 gp41) with receptors CD4, CXCR4 and/or CCR5 followed by fusion of viral and host membranes. Studies indicate that additional factors such as receptor density and composition of viral and cellular lipids can dramatically modulate the fusion reaction. Lipid rafts, which primarily consist of sphingolipids and cholesterol, have been implicated for infectious route of HIV-1 entry. Plasma membrane Glycosphingolipids (GSLs) have been proposed to support HIV-1 infection in multiple ways: (a) as alternate receptor(s) for CD4-independent entry in neuronal and other cell types, (b) viral transmission, and (c) gp120-gp41-mediated membrane fusion. However, the exact mechanism(s) by which GSLs support fusion is still elusive. This article will focus on the contribution of target membrane sphingolipids and their metabolites in modulating viral entry. We will discuss the current working hypotheses underlying the mechanisms by which these lipids promote and/or block HIV-1 entry. Recent approaches in the design and development of novel glycosyl derivatives, as anti-HIV agents will be summarized. PMID- 16307381 TI - Cloning and characterization of an agglutinin gene from Arisaema lobatum. AB - A novel agglutinin gene was cloned from Arisaema lobatum using SMART RACE-PCR technology. The full-length cDNA of Arisaema lobatum agglutinin (ala) was 1078 bp and contained a 774 bp open reading frame encoding a lectin precursor (proproprotein) of 258 amino acid residues with a 23 aa signal peptide. ALA contained three mannose-binding sites (QXDXNXVXY) with two-conserved domains of 45% identity, ALA-DOM1 and ALA-DOM2. The three-dimensional structure of ALA was very similar to that of GNA (Galanthus nivalis agglutinin). ALA shared varying identities, ranging from 40% to 85%, with mannose-binding lectins from other species of plant families, such as Araceae, Alliaceae, Iridaceae, Lillaceae, Amaryllidaceae and Bromeliaceae. Genomic sequence of ala was also cloned using genomic walker technology, and it was found to contain three putative TATA boxes and eight possible CAAT boxes in the 5'-flanking region. No intron was found within the region of genomic sequence. Southern blot analysis indicated that the ala belonged to a multi-copy gene family. Expression pattern analysis revealed that the ala preferentially expressed in the tissues with the higher expression being found in spadix, bud, leaf, spathe and tuber. The cloning of the ala gene not only provides a basis for further investigation of its structure, expression and regulation mechanism, but also enables us to test its potential role in controlling pests and fungal diseases by transferring the gene into plants in the future. PMID- 16307382 TI - Transmembrane signaling in the brain by serotonin, a key regulator of physiology and emotion. AB - Serotonin (5-HT) is an ancient chemical that plays a crucial functional role in almost every living organism. It regulates platelet aggregation, activation of immune cells, and contraction of stomach and intestinal muscles. In addition, serotonin acts as a neurotransmitter in the brain and the peripheral nervous system. These activities are initiated by the binding of serotonin to 15 or more receptors that are pharmacologically classified into seven groups, 5-HT1 through 5-HT7. Each group is further divided into subgroups of receptors that are homologous but are encoded by discrete genes. With the exception of the 5-HT3 receptor--a cation channel--all of the others are G protein-coupled receptors that potentially activate or inhibit a large number of biochemical cascades. This review will endeavor to compare and contrast such signaling pathways with special attention to their tissue-specific occurrence, their possible role in immediate effects on covalent modification of other proteins, and relatively slower effects on gene expression, physiology and behavior. PMID- 16307383 TI - Cytochrome C effect on respiration of heart mitochondria: influence of various factors. AB - The effect of exogenous cytochrome c on respiration rate of the rat and human heart mitochondria was assessed in situ, using permeabilized fibers. It was (i) much more pronounced in State 2 and 4 than in State 3 with all the respiratory substrates (pyruvate+malate, succinate, palmitoyl-CoA+carnitine and octanoyl-L carnitine), (ii) different with different substrates, (iii) much higher after ischemia in both metabolic states, particularly in the case of succinate oxidation compared to pyruvate+malate, (iv) the highest in State 4 with succinate as a substrate. Similar results were obtained with the isolated rat and rabbit heart mitochondria. The differences in the degree of stimulation of mitochondrial respiration by cytochrome c and, thus, sensitivity of cytochrome c test in evaluation of the intactness/injury of outer mitochondrial membrane are probably determined by the differences in the cytochrome c role in the control of mitochondrial respiration in the above-described conditions. PMID- 16307384 TI - DGGE fragments oscillate with or counter to fluctuations in cultivable bacteria along wheat roots. AB - Previously, we showed that bacterial populations oscillate in response to a moving substrate source such as a root tip, resulting in moving wavelike distributions along roots. For this article, we investigated if bacterial communities fluctuate as a whole or if there is a succession in bacterial composition from peak to peak or within peaks. Rhizosphere microbial communities along roots of wheat Triticum aestivum L. were studied in detail (20-25 rhizosphere and bulk soil samples along the total root length) in two related soils by colony enumeration and culture-independent DNA analysis. Similar to our previous findings, the numbers of copiotrophic and oligotrophic bacteria oscillated with significant harmonics along each root, independent of soil moisture or lateral roots. Shifts in amplified eubacterial 16S rDNA fragments from denaturing gradient gel electrophoresis (DGGE) analysis were detected along the roots. The most abundant and intensively amplified fragments fluctuated in phase with colony-forming unit (CFU) oscillations; fewer amplified fragments with less intensive bands fluctuated out of phase or were restricted to certain root zones. The bacterial species richness along the root was negatively correlated with the numbers of oligotrophic bacterial CFUs. Discriminant analyses on DGGE patterns distinguished between soil types, rhizosphere and bulk soil, and waxing and waning phases in the oscillations along roots. Bacterial compositions shifted within oscillations but were repeated from oscillation to oscillation, supporting the idea that the most abundant bacterial taxa were growing and dying over time and consequently in space, whereas other taxa counterfluctuated or hardly responded to the substrate supplied by the passing root tip. PMID- 16307385 TI - Kinetics of insulin adsorption at the oil-water interface and diffusion properties of adsorbed layers monitored using fluorescence correlation spectroscopy. AB - The adsorption of insulin at an oil-water interface was studied with fluorescence correlation spectroscopy (FCS). FCS is able to measure diffusion properties of insulin at nanomolar concentrations, making it possible to detect the very early steps in the adsorption process. Below 20 nM bulk insulin concentration, the insulin molecules adsorbed to the surface diffuse freely at all times during the experiment (a few hours). At higher concentrations, a surprisingly abrupt transition to a slow diffusion phase is observed. Based on the information about both diffusion times and molecular brightness derived from the FCS experiments, we suggest that the transition represents the formation of a fractal network. FCS may be a valuable tool in pharmaceutical formulation science, because it provides information about concentration buildup and phase changes at interfaces formed in drug delivery systems. PMID- 16307386 TI - Nanoparticles of poorly water-soluble drugs prepared by supercritical fluid extraction of emulsions. AB - PURPOSE: The aim of the study was to develop and evaluate a new method for the production of micro- and nanoparticles of poorly soluble drugs for drug delivery applications. METHODS: Fine particles of model compounds cholesterol acetate (CA), griseofulvin (GF), and megestrol acetate (MA) were produced by extraction of the internal phase of oil-in-water emulsions using supercritical carbon dioxide. The particles were obtained both in a batch or a continuous manner in the form of aqueous nanosuspensions. Precipitation of CA nanoparticles was used for conducting a mechanistic study on particle size control and scale-up. GF and MA nanoparticles were produced in several batches to compare their dissolution behavior with that of micronized materials. The physical analysis of the particles produced was performed using dynamic light scattering (particle size), scanning electron microscopy (morphology), powder X-ray diffraction (crystallinity), gas chromatography (residual solvent), and a dissolution apparatus. RESULTS: Particles with mean volume diameter ranging between 100 and 1000 nm were consistently produced. The emulsion droplet size, drug solution concentration, and organic solvent content in the emulsion were the major parameters responsible for particle size control. Efficient and fast extraction, down to low parts-per-million levels, was achieved with supercritical CO2. The GF and MA nanoparticles produced were crystalline in nature and exhibited a 5- to 10 fold increase in the dissolution rate compared with that of micronized powders. Theoretical calculations indicated that this dissolution was governed mainly by the surface kinetic coefficient and the specific surface area of the particles produced. It was observed that the necessary condition for a reliable and scalable process was the sufficient emulsion stability during the extraction time. CONCLUSION: The method developed offers a viable alternative to both the milling and constructive nanoparticle formation processes. Although preparation of a stable emulsion can be a challenge for some drug molecules, the new technique significantly shortens the processing time and overcomes the current limitations of the conventional precipitation techniques in terms of large waste streams, product purity, and process scale-up. PMID- 16307387 TI - A clinical and molecular overview of the human osteopetroses. AB - The osteopetroses are a heterogeneous group of bone remodeling disorders characterized by an increase in bone density due to a defect in osteoclastic bone resorption. In humans, several types can be distinguished and a classification has been made based on their mode of inheritance, age of onset, severity, and associated clinical symptoms. The best-known forms of osteopetrosis are the malignant and intermediate autosomal recessive forms and the milder autosomal dominant subtypes. In addition to these forms, a restricted number of cases have been reported in which additional clinical features unrelated to the increased bone mass occur. During the last years, molecular genetic studies have resulted in the identification of several disease-causing gene mutations. Thus far, all genes associated with a human osteopetrosis encode proteins that participate in the functioning of the differentiated osteoclast. This contributed substantially to the understanding of osteoclast functioning and the pathogenesis of the human osteopetroses and will provide deeper insights into the molecular pathways involved in other bone pathologies, including osteoporosis. PMID- 16307388 TI - Recommendations for thresholds for cortical bone geometry and density measurement by peripheral quantitative computed tomography. AB - Peripheral quantitative computed tomography (pQCT) is widely used for clinical and research purposes. For accurate determination of bone geometry (bone cross sectional area, cortical thickness, and cortical area), volumetric bone mineral density (vBMD) and cortical bone mineral content (BMC), it is important to select the appropriate thresholds. A Stratec XCT-2000 scanner was used to compare current standard practice with new optimized thresholds. Currently, a single threshold of 710 mg/mL for the measurement of cortical vBMD and geometry is used. We hypothesised that this threshold may not be optimal and used the European Forearm Phantom (EFP) and patient data to test more appropriate thresholds. A single slice (1.2 mm width, 0.4 mm pixel size) was made at section 4 of the EFP (representing the diaphyseal portion of a long bone). The EFP has a known cortical thickness of 2.5 mm and, therefore, the correct threshold for geometry would be that which measures cortical thickness as 2.5 mm. Thresholds were altered at approximately the 50% value between soft tissue (60 mg/mL) and peak density (879 mg/mL), and cortical thickness versus threshold was plotted; the correct threshold for geometry was 460 mg/mL. By expressing this threshold as a percentage of the range of density values in the EFP ([460-60]/[879-60] = 49%) and then applying this percentage to in vivo data, the optimum threshold for geometry can be determined: ([1240-79] x 0.49) + 79 = 648 mg/mL. For cortical vBMD of in vivo bone measurements at the midshaft site of the radius, thresholds were varied around the peak value (1240 mg/mL), and the threshold was set to that which gave a cortical density of 1240 mg/mL; the threshold for cortical density was, therefore, 1200 mg/mL. A subset of radius scans from a population of young healthy females was analyzed using the new thresholds (648 mg/mL for bone geometry, 1200 mg/mL for cortical vBMD) versus the current threshold (710 mg/mL). For bone geometry, the mean difference between the analysis based on the new threshold and that based on the manufacturer-recommended threshold ranged between 2.1% and 14% (total area = 2.1%, cortical thickness = 14%, cortical area = 3.7%). Although there was a 10% difference between the analysis based on the new threshold and that based on the manufacturer-recommended threshold, this difference was not systematic. Thresholds will significantly affect results obtained from pQCT. The current threshold of 710 mg/mL is inadequate for accurate determination of bone geometry and cortical vBMD. New thresholds of 648 mg/mL for geometry and 1,200 mg/mL for cortical vBMD should be used. PMID- 16307389 TI - Differences in osteocyte density and bone histomorphometry between men and women and between healthy and osteoporotic subjects. AB - Bone defects related to osteoporosis develop with increasing age and differ between males and females. It is currently thought that the bone remodeling process is supervised by osteocytes in a strain-dependent manner. We have shown an altered response of osteocytes from osteoporotic patients to mechanical loading, and osteocyte density is reduced in osteoporotic patients, which might relate to imperfect bone remodeling, leading to lack of bone mass and strength. Hence, information on osteocyte density will contribute to a better understanding of bone biology in males and females and to the assessment of osteoporosis. Osteocyte density as well as conventional histomorphometric parameters of trabecular bone were determined in cancellous iliac crest bone of healthy postmenopausal women and men and of osteoporotic women and men. Osteocyte density was higher in healthy females than in healthy males and lower in osteoporotic females than in healthy females. Bone mass was reduced in osteoporotic patients, both male and female. In females, trabecular number was reduced, whereas in males, trabecular thickness was reduced and eroded surface was increased. There were no correlations between the parameter groups bone architecture, bone formation, bone resorption, and osteocyte density. These results are consistent with impaired osteoblast function in osteoporotic patients and with a different mechanism of bone loss between men and women, in which osteocyte density might play a role. The reduced osteocyte numbers in female osteoporotic patients might relate to imperfect bone remodeling leading to lack of bone mass and strength. PMID- 16307390 TI - Prevention of bone loss by phloridzin, an apple polyphenol, in ovariectomized rats under inflammation conditions. AB - Aging and sex hormones related changes lead to inflammatory and oxidant conditions, which are involved in the pathogenesis of osteoporosis. Recent studies have suggested that polyphenols may exert a protective effect in such conditions. We assessed the effect of phloridzin (Phlo), a flavonoid exclusively found in apple, on bone metabolism in ovariectomized (OVX) or sham-operated (SH) rats with and without inflammation. Six-month-old Wistar rats were allocated to two equal groups that received either a control diet or a diet supplemented with 0.25% Phlo for 80 days. Three weeks before necropsy, inflammation was induced by subcutaneous injection of talc in 10 animals of each group. At necropsy, ovariectomy decreased both total (T-BMD) and metaphyseal (M-BMD) femoral bone mineral density (P < 0.01). Inflammation conditions, checked by an increase in the spleen weight and alpha1-acid glycoprotein concentration in OVX rats, exacerbated the decrease in T-BMD (g/cm2) (as well as M-BMD) observed in castrated animals (P < 0.05). Daily Phlo intake prevented ovariectomy-induced bone loss in conditions of inflammation as shown by T-BMD and M-BMD (P < 0.05). At the diaphyseal site, BMD was improved by Phlo in OVX rats with or without inflammation (P < 0.05). These results could be explained by changes in bone remodeling as the increased urinary deoxypyridinoline excretion in OVX and OVXinf animals was prevented by the polyphenol-rich diet (P < 0.001), while plasma osteocalcin concentration was similar in all experimental groups. In conclusion, Phlo consumption may provide protection against ovariectomy-induced osteopenia under inflammation conditions by improving inflammation markers and bone resorption. PMID- 16307391 TI - Bone biomechanical property deterioration due to tobacco smoke exposure. AB - Tobacco smoking has been implicated in the development of osteoporosis and early onset of menopause in women smokers. We measured various biomechanical properties of femurs and tibiae obtained from smoke-exposed and control mice to determine cigarette smoke influences on bone mass, structure, and strength. Growing female C57BL mice were exposed to sidestream cigarette smoke in a whole-body exposure chamber, set at 30 +/- 2 mg smoke particulates/m3 for 4 hours/day and 5 days/week for 12 consecutive weeks. Elevated levels of urinary cotinine and pulmonary ethoxyresorufin deethylase activity in smoke-exposed mice confirmed their effective exposure to cigarette smoke. There were no differences in body weight and physical size (length, medial-lateral and anterior-posterior widths, midshaft cortical area and thickness) of femurs and tibiae between smoke-exposed and control mice. The femoral mid-shaft yield load, stiffness, yield stress, and modulus were, respectively 8%, 13%, 10%, and 14% lower (P < 0.05) in smoke exposed compared to control mice. The ultimate load and stress in mid-shaft femurs showed decreasing trends (P < 0.1) in smoke-exposed mice. In the femoral neck, the ultimate load and stiffness were 9% and 12% lower (P < 0.05) in smoke exposed mice, respectively. Further, the ash-to-dry bone weight ratio was smaller ( approximately 6%, P < 0.05), and micro-computed tomographic scanning of distal femoral bone volume/total volume (%) and trabecular thickness showed decreasing trends in smoke-exposed mice compared to the control group. We conclude that exposure to tobacco smoke deteriorates some of the biomechanical properties of bone in growing female mice. PMID- 16307392 TI - Osteocyte apoptosis and osteoclast presence in chicken radii 0-4 days following osteotomy. AB - Osteocyte apoptosis caused by load-induced microdamage is followed by osteoclastic bone remodeling, and a causal link between apoptosis and repair has been suggested. The objectives of the present study were to use a chick model to examine the incidence of osteocyte apoptosis and the presence of osteoclasts during the first 96 hours following an osteotomy, prior to extensive callus mineralization. Osteotomies were performed on the right radii of 24 chicks at 23 24 days of age. The left radii served as controls. Radii were collected and processed at six time points following surgery (0, 12, 24, 48, 72, and 96 hours). Decalcified bone tissue sections were stained either for apoptosis using a modified TUNEL procedure or for tartrate-resistant acid phosphatase to identify osteoclasts in the intracortical and periosteal envelopes. The percentage of apoptotic osteocytes, as well as osteoclast counts (n/mm or n/mm2) were quantified in four regions (0-1, 1-2, 2-4, and 4-8 mm from the site of the osteotomy; regions 1-4, respectively) in the osteotomized radii and in the same measured areas in the control radii. Data for osteocyte apoptosis and osteoclasts in the control limb were subtracted from the osteotomized limb data to identify differences due to surgical influence. The incidence of osteocyte apoptosis was significantly higher at 12, 24, 48, and 72 hours versus 0 hours following osteotomy, and the response was highest in region 1; however, there was no interaction between time and region. Intracortical osteoclast counts (n/mm2) were elevated after 48 hours, and the response was similar in all regions. The data demonstrate that osteocyte apoptosis occurs within 24 hours in response to an osteotomy and temporally precedes an increase in osteoclast presence. Hence, osteocyte apoptosis may play a role in signaling during the bone healing process. PMID- 16307394 TI - Lag screw fixation in midface fractures. AB - Through analysis of case series from medical record review, this article demonstrates how lag screw fixation may be used to effectively treat obliquely oriented maxillary, zygomatic, and orbital rim fractures. Lag screw fixation of midface fractures has been overshadowed by other readily available plate fixation techniques. Nonetheless, lag screw fixation provides quick, stable, and effective reduction of obliquely oriented maxillary, zygomatic, and orbital fractures without risk of plate exposure and with potentially superior primary osteosynthesis. Lag screw fixation remains an excellent alternative to plate fixation techniques in the repair of appropriate midface fractures. PMID- 16307395 TI - Transconjunctival approach to the zygomatico-frontal limb of orbitozygomatic complex fractures. AB - Fractures of the orbitozygomatic complex represent one of the most common injuries of the facial skeleton. Repair of the fractures often requires multiple surgical access points and the necessity for skin incisions. As surgeons attuned to aesthetics, we strive to minimize the signs of injury and treatment. We propose an approach designed to treat orbitozygomatic fractures without the use of skin incisions. The objective of this article is to outline the surgical technique designed by the senior author (PRL) to address the zygomatico-frontal fracture component of orbitozygomatic complex fractures through an upper lid lateral transconjunctival approach. Relevant anatomy and technique is described. The combination of previously described lower lid transconjunctival and gingivobuccal sulcus incisions with this new upper lid lateral transconjunctival approach provides adequate access for rigid fixation of all major components of orbitozygomatic fractures without skin incisions. PMID- 16307396 TI - Operative time analysis of miniplate fixation of the mandible. AB - Our purpose in this study was to report and compare operating room times for the various methods of mandible fracture repair. We describe a methodology of treatment in selected patients, using efficient repair techniques; namely, miniplate fixation with four-screw mandibular occlusion. We report our outcomes in successful healing, occlusion, complications, and operating times. A retrospective chart review was performed of 68 patients suffering mandible fractures treated by various surgeons at a single institution. Miniplate fixation technique requires significantly less time in the operating room than other mandible repair techniques, including mandibulomaxillary fixation. Miniplate fixation technique is both an efficacious and an efficient means of repairing certain mandible fractures. PMID- 16307397 TI - Anterior skull base fractures. AB - Anterior skull base trauma evaluation and management has historically been difficult to systematically study secondary to the relative rarity of its occurrence, associated major morbidity and high mortality, and poor radiographic techniques. It has only been recently that improved care has allowed for decreased morbidity and mortality. The advent of computed tomography imaging techniques has led to anatomic characterization and detailed study of injury patterns. This article reviews current techniques for evaluation and management of the associated sinonasal, orbital, and neurologic sequelae of anterior skull base injuries. PMID- 16307398 TI - Combined laryngotracheal separation and esophageal injury following blunt neck trauma. AB - In this article we describe the first case of combined laryngotracheal and esophageal injury following blunt neck trauma in the otolaryngology literature and delineate appropriate management considerations. Successful reconstruction requires appropriate airway management, restoration of esophageal and laryngeal mucosal integrity, and reduction of tracheal and laryngeal fractures with appropriate stenting. Laryngotracheal separation following blunt neck trauma is rare and can be initially overlooked. After the airway is secure, injury severity dictates repair and postoperative care. Immediate reconstruction with restoration of the laryngotracheal framework and mucosal integrity enables patients to recover a patent airway, functional voice, and normal swallow. PMID- 16307399 TI - Gunshot wounds to the face--acute management. AB - The complex facial trauma victim poses a genuine therapeutic challenge as a whole, and may be particularly challenging to the medical team. The literature on acute management of gunshot wounds to the face is scarce. We performed an extensive review of the English-language literature in an effort to better delineate the diagnosis and acute management of these injuries. Most of these injuries do not present with initial threat to life and can safely be managed non operatively. Definitive treatment is often deferred in patients with multiple, or more severe, injuries. Airway compromise is the most frequent and most life threatening early problem reported in most series. CT scan remains the most useful method in the evaluation of these types of injuries and associated cervical spine lesions. Facial bleeding is best controlled by angiography and subsequent embolization. Anatomic repair of soft tissue and bony injuries is recommended to obtain an optimal functional and aesthetic outcome. Despite the creation of some algorithms, no clear correlation has been found between the site of entrance wound and the injuries and outcome of gunshot wounds to the face. PMID- 16307400 TI - Frontal sinus fractures: management guidelines. AB - Management of frontal sinus fractures (FSF) has been the subject of great debate for more than six decades. Multiple treatment options and algorithms have been proposed by multiple specialties throughout the years; however, the optimal method of frontal sinus repair has yet to be elucidated. Because of the location of the frontal sinus and its proximity to numerous intracranial structures, inadequate treatment may lead to life-threatening intracranial infectious complications. Meningitis, encephalitis, and brain abscess are the most common intracranial complications. Other complications include persistent cerebrospinal leakage, mucopyoceles, frontal osteomyelitis, meningoencephalocele, and nonunion of the frontal bone. Orbital involvement may result in ophthalmoplegia, orbital abscess, diplopia, enophthalmos, proptosis, preseptal cellulitis, and partial or complete loss of vision. Morbidity and mortality are often dependent on the anatomic characteristics of the fracture, concomitant injuries, treatments rendered, age, gender, and mechanism of injury. Management of frontal sinus fractures is so controversial that the indications, timing, method of repair, and surveillance remain disputable among several surgical specialties. The most important tenet of frontal sinus fracture management remains the same: create a safe sinus. This is accomplished by following four basic principles: reestablish the frontal bony contour to its premorbid state, restore normal sinus mucosa with a patent drainage system if possible, eradicate the sinus cavity if the normal mucosa or drainage system cannot be reestablished, and create a permanent barrier between the intracranial and extracranial systems to prevent overwhelming infectious complications. By following these four basic principles, frontal sinus fracture management will be safe and effective as long as extended surveillance is part of the protocol. PMID- 16307401 TI - Orbital floor fracture management. AB - Orbital floor fractures can occur in isolation or in conjunction with other facial skeletal fractures. They are commonly associated with midface fractures and, in this region, are second only to nasal fractures in occurrence. The integrity of the orbit and its contents must be preserved to prevent complications after alterations secondary to trauma. There is a plethora of literature on different reconstruction methods. Autogenous bone and cartilage grafts from various areas have been used in the reconstruction of the floor. In addition, materials such as high-density porous polyethylene, silicone, and titanium mesh have also been used. Each has its own benefits and limitations. The lack of stabilization and improper posterior placement of the reconstruction implant have caused less-than-ideal repairs. Graft stabilization can be accomplished with titanium screw fixation, and recently the endoscope has facilitated the visualization of the entire defect and helped ensure the proper placement of the implant. PMID- 16307402 TI - Use of three-dimensional computerized tomography reconstruction in complex facial trauma. AB - Technological advances in computerized tomography (CT) have reduced data acquisition and reconstruction times so that three-dimensional (3D) CT images of maxillofacial injuries may be economically and quickly generated. 3DCT was judged superior to multiplanar two-dimensional CT in demonstrating the spatial relationships of fracture fragments in complex mandibular and midfacial trauma. Although 3DCT failed to demonstrate soft-tissue injuries well, the surgeon's improved appreciation of the disrupted bony architecture facilitated preoperative planning. 3DCT facilitates the evaluation of complex mandibular and midfacial fractures. PMID- 16307403 TI - [Does migration from high and intermediate endemic regions increase the prevalence of hepatitis B infection in Germany?]. AB - BACKGROUND AND OBJECTIVE: Germany is one of the low endemic areas for hepatitis B. There are 7,3 million foreign citizens and 3,2 million migrants from the former USSR and Eastern Europe with German roots, the "Resettlers" (Aussiedler), who migrated to Germany mostly from countries with moderate or high HBsAg prevalence. The aim of this study was to determine the HBsAg prevalence in adult foreign citizens and resettlers compared with that among the adult German population. METHODS: Adult foreign citizens and resettlers were categorized according to their country of origin into groups with low, intermediate or high HBsAg prevalence, using data from the WHO. Statistics of the Federal Office for Statistics, the Federal Office for Administration and the Federal Ministry of the Interior were used for the demographic analysis. The number of chronic HBsAg carriers for the different population groups and the whole population was then calculated. RESULTS: 84% of the adult migrant population in Germany migrated from countries with intermediate and high HBsAg endemicity. For 2003 we calculated 503 040 HBsAg carriers in Germany. 42% of these have a migratory background, even though migrants represent only 12.7% of the whole population. The risk for chronic infection with HBsAg is 7.1 (4.8-13.2) for the resettlers and is 4.3 (3.0 8.1) times higher for foreign citizens than for the German population. CONCLUSIONS: These remarkable differences in prevalence make it important that migrants and their close contacts be vaccinated properly, pregnant women be included in screening programs and cases of chronic hepatitis B be treated adequately. PMID- 16307404 TI - [Resuscitation by laypersons: lack of knowledge of first-aid measures in Germany]. AB - BACKGROUND AND OBJECTIVE: Sudden cardiac death is a leading cause of death. In Germany, only 10% of patients will survive reanimation. The prognosis is mainly determined by the first-aid skills of accidental witnesses. METHODS: The reaction of 1007 German-speaking adults (462 males, 545 females, median age 39 years) was investigated in an emergency scenario with symptoms of sudden cardiac death in an acquaintance. Afterwards a self-assessment of their first-aid skills, such as cardiac compression and mouth-to-mouth ventilation was made. RESULTS: 94% of the interviewees would call professional aid at once. Diagnostic skills, as searching for pulse or checking for breath would be done by 26 and 21%, respectively. The most frequently mentioned therapeutic skill was positioning the victim in stable lateral position (37%), but very less frequently cardiac compression (6%) and mouth-to-mouth resuscitation (10%). When asked, 51 (81%) of those interviewed regarded their skills in cardiac compression and mouth-to-mouth resuscitation as adequate. To some extent there were differences in favor of young people and those from East Germany. CONCLUSION: These representative data suggest that (a) ABC-like first-aid rules are largely unknown among the population; (b) the value of a stable lateral position is overestimated and (c) first-aid skills of elderly persons are worst. A significant improvement of first-aid skills of the German population is mandatory in order to improve in future the outcome of sudden cardiac death. PMID- 16307406 TI - [Curative treatment in a patient with exercise induced syncope]. AB - HISTORY: A 38 year old man suffered from exercise-induced reproducible dizziness and syncopes. INVESTIGATIONS AND DIAGNOSIS: During exercise testing a ventricular tachycardia at a rate of 300 beats/min was identified as the cause of the symptoms. Because of the ventricular morphology with inferior axis, left bunde branch block and the typical monomorphic repetitive characteristics, idiopathic adenosine-sensitive ventricular tachycardia was diagnosed. TREATMENT AND CLINICAL COURSE: Curative catheter ablation of the arrhythmogenic focus in the right ventricular outflow tract was performed. The patient has now been free of symptoms for more than two years. The characteristics of idiopathic ventricular tachycardia and the electrophysiological techniques are described. CONCLUSION: Idiopathic ventricular tachycardia is a rare cause of syncope in young patients without underlying heart disease and can be cured by catheter ablation. Exclusion of cardiac diseases, especially arrhythmogenic right ventricular cardiomyopathy, is of prognostic value. PMID- 16307407 TI - [Fever of unknown origin in the 21st century. 2. Non-infectious diseases (autoimmune diseases)]. PMID- 16307408 TI - [Reflux-associated sleep disorders]. AB - Difficulties of initiating or maintaining sleep are very common in the general population. Individuals with clinical sleep disorders have a greatly impaired quality of life. The causes for sleeping disorders are complex, but evidence has recently come from different trials supporting a causal relationship between gastro-esophageal reflux disease (GERD) and sleep disorders in some patients. The majority of patients with GERD report reflux symptoms during the night. It is well known that especially at night reflux is characterized by prolonged esophageal acid exposure. Recent data have indicated that sleep disorders significantly improve while on efficacious antisecretory treatment. In particular in patients with sleep disorders but no previously known GERD, the search for it is recommended and should be followed by adequate antisecretory treatment. In other severe diseases associated with sleep, like the obstructive sleep apnoea syndrome (OSAS), an association with esophageal acid exposure has been proven. The sleep apnea-associated reflux has probably a multifactorial etiology: in cases with other predisposing conditions for gastro-esophageal reflux, OSAS promotes the development of reflux. Antisecretory treatment has been able to improve the severity of OSAS. The purpose of this review has been to survey of recent publications in GERD-associated sleep disorders, as well as the possible pathophysiological basis of the observed phenomena. PMID- 16307409 TI - [Innovative antibody-therapies for malignant lymphomas]. AB - Non-Hodgkin s lymphoma (NHL) includes a group of malignant lymphoproliferative disorders, that particularly occur in the elderly and have an continuously increasing incidence. Because of the age distribution new treatment options with low toxicity and minor side effects - apart from "conventional" therapies like standard or high dose chemotherapy - are needed. Within these novel therapeutic modalities the use of the monoclonal antibody rituximab has been widely established. Other monoclonal antibodies, such as the anti-CD52-antibody alemtuzumab, are available for the treatment of chronic lymphatic leukemia (CLL) and T-cell-lymphomas and are being tested in clinical trials. Furthermore the option of combining targeted therapies, in which antibodies are used with radiotherapy, has led to the development of radio-immunotherapies that are now becoming available for clinical use. Current results in the treatment of advanced indolent lymphomas are promising, and their use in high-grade lymphoma and as first-line therapy is under investigation. This article summarizes the application of these novel immunotherapies and reviews the results of recent clinical trials, with particular emphasis on indications and practical aspects in everyday clinical life. PMID- 16307410 TI - [Where and in which blood vessels does atherosclerosis manifest itself?]. PMID- 16307412 TI - [Calculated renal function: benefits and problems of the "Modification of Diet in Renal Disease" (MDRD) formula]. PMID- 16307414 TI - [Osteoporosis in patients during oral anticoagulant treatment]. PMID- 16307415 TI - [One group ticket "from bench to bedside", return, please!]. PMID- 16307416 TI - Prognostic impact of age in children and adolescents with acute lymphoblastic leukemia: data from the trials ALL-BFM 86, 90, and 95. AB - Large progress has been made in the treatment of acute lymphoblastic leukemia (ALL) of childhood and adolescence over the past 30 years. Eighty percent of the patients can be cured, but clinical subgroups with a dismal outcome can still be identified. In this study, we investigated the association of age with prognosis in 5 181 patients with ALL under 18 years (y) of age enrolled in the three consecutive treatment trials ALL-BFM 86, 90 and 95 in more than 80 centers. Event free survival (pEFS) of the total group was significantly associated with age. The most unfavorable outcome was found in infancy and the best results were achieved at toddler and pre-school age. Beyond 5 y of age, survival probability decreased (pEFS at 8 y: < 1 y = 0.45; 1-5 y = 0.82; 6-9 y = 0.75; 10-14 y = 0.63; > or = 15 y = 0.59). The proportion of T-ALL as compared to precursor B-cell ALL (pB-ALL) was lower in younger children, due to an incidence peak of pB-ALL in toddlers and at pre-school age compared to a constant incidence of T-ALL. Within the T-ALL group, no correlation of age with sex, initial white blood cell count, CNS disease, or early treatment response was found. Children under 10 y of age had a slightly lower relapse rate compared to older patients. Within pB-ALL patients, the proportion as well as the absolute incidence of TEL/AML1 rearrangement and DNA index of > or = 1.16 was higher in the younger children. A lower proportion of BCR/ABL-positive ALL was observed in the age group of < 6 y when compared to patients aged > or = 6 y, but the absolute incidence was constant across the age groups after the first year of life. More than half of the infants had a CD10-negative pB-ALL. The incidence was constant after a peak in the first year of life, yet the percentage of CD10 negativity increased with rising age in this subgroup. Adolescents with pB-ALL had a significantly higher proportion of prednisone poor-responders. Accordingly, outcome was worse in older patients. This pattern was also evident in the biologically heterogeneous group of patients with a DNA index of > or = 1.16. In contrast, no significant age related outcome differences could be shown within TEL/AML1- or BCR/ABL-positive patients, as well as within CD10-negative pB-ALL beyond infant age. Analysis of the pB-ALL group in a Cox's regression model including age and the above-listed biological factors revealed age < 1 year and > or = 10 years as independent risk factors. This is in line with the poorer prognosis of these age groups in the pB ALL subgroup without specific biological characteristics. This subgroup also had an incidence peak at toddler age, presumably containing other favorable biological subsets. An independent prognostic impact of age in pediatric ALL cannot be excluded by this study. However, our analyses show that the age associated different prognosis in childhood ALL is at least partly related to the different distribution of relevant prognostic subgroups between the age groups. PMID- 16307417 TI - Asparagine concentration in plasma after 2,500 IU/m(2) PEG-asparaginase i.v. in children with acute lymphoblastic leukemia. AB - Polyethylene glycol conjugated asparaginase (PEG-ASNase) can be substituted in cases of hypersensitivity to native Escherichia coli asparaginase. We measured asparagine (asn) levels in plasma after a single dose of 2,500 IU/m(2) i.v. PEG ASNase (Oncaspar) in consolidation treatment of ALL and compared those with data from the previous protocol COALL-05-92. This protocol was similar to COALL-06-97, except that children had been given 45,000 IU/m(2) C-ASNase instead of PEG ASNase. PATIENTS AND METHODS: Between May 2000 and December 2001 seventy-one children (38 boys, 33 girls) with newly diagnosed ALL treated according to the multicenter protocol COALL-06-97 were investigated in this study. Four hundred and seventy-four plasma samples (71 patients) were analysed by ion exchange chromatography after column derivatization with o-phthaldialdehyde. For comparison data (350 plasma samples) from 51 patients treated according to the protocol COALL-05-92 were available. The same method for detection of asn in plasma was used. RESULTS: The median asparagine level in plasma after 2,500 IU/m(2) PEG-ASNase i.v. was below the limit of detection for at least 5 weeks in 81 % of the patients. When divided into high risk (HR) and low risk (LR) group, HR patients who had previously received one dose more of C-ASNase showed a markedly shorter depletion than the LR patients compatible with a higher risk of antibody formation and consequent silent inactivation after a higher number of exposures to ASNase. In the previous protocol COALL-05-92 median asn levels in plasma after 45,000 IU/m(2) native C-ASNase i.v. were below the limit of detection for at least 5 weeks in 65 % of the patients. CONCLUSIONS: 2,500 IU/m(2) PEG-ASNase led to an equally long depletion of asn in plasma as did 45,000 IU/m(2) native C-ASNase i.v. used in COALL-05-92. PMID- 16307418 TI - FDG-PET in 10 children with non-Hodgkin's lymphoma: initial experience in staging and follow-up. AB - BACKGROUND: The use of FDG-PET was evaluated for initial staging and therapy efficacy in paediatric patients with non-Hodgkin's lymphoma (NHL) and compared to the established conventional imaging modalities (CIM). The results of this retrospective analysis are presented in conjunction with a critical review of the current literature. PATIENTS AND METHODS: Ten paediatric patients with NHL were examined using whole-body FDG-PET initially (n = 6), during therapy (n = 5) and after completion of therapy (n = 5), respectively. FDG-PET findings were compared to CIM performed according to the protocol of the German NHL-BFM 95 study. The results were evaluated for their impact on disease classification and therapy decision (St. Jude, REAL) in correspondence to a clinical follow-up of at least 24 months. RESULTS: Concerning initial staging, all lymphoma manifestations detected by conventional imaging were also detected by FDG-PET (15 nodal, 2 extranodal). Furthermore, an additional nodal lesion was detected by FDG-PET in three patients. This resulted in an upstaging followed by an intensified poly chemotherapy in one patient. In five patients showing unclear residual masses on conventional imaging during therapy, FDG-PET indicated viable residual tumours in one case. This patient showed a relapse during follow-up while the four FDG-PET negative patients did not. After completion of initial therapy, FDG-PET revealed in one out of five patients persistent tumour metabolism in the primary lesions and also detected new manifestations. The patient died shortly after restaging due to disease progression. CONCLUSIONS: These first results on the use of FDG PET in paediatric non-Hodgkin lymphoma indicate a high potential to improve the therapeutic management. PMID- 16307419 TI - A comparison between three graft manipulation methods for haploidentical stem cell transplantation in pediatric patients: preliminary results of a pilot study. AB - Transplantation of hematopoietic stem cells from mismatched related donors makes a potential donor available for every child in need of stem cell transplantation. Here, we compare three different graft manipulation methods in patients with leukemias and lymphomas: positive selection of stem cells with either CD34 (n = 39) or CD133-coated magnetic microbeads (n = 14) and a new strategy which depletes T- and B-cells through the use of CD3- and CD19-coated microbeads (n = 11). Median purity of stem cells was comparable after CD34 (+)-selection and CD133 (+)-selection, whereas stem cells were only slightly enriched after CD3 (+)/CD19 (+)-depletion (97.5 %, 93.4 % and 1.02 %). Indirect depletion of T-cells by positive selection resulted in 1 x 10 (4) (median) residual CD3 (+)-cells/kg (0.7-3 x 10 (4)). Patients with CD3/CD19-depleted grafts received 3.2 x 10 (4) (median) (0.7-16 x 10 (4)) residual T-cells/kg. Those grafts also comprised NK cells (median number: 86 x 10 (6)/kg), dendritic cells and monocytes/granulocytes. Primary engraftment of the stem cell products was comparable after CD34- and CD133-selection (85 and 72 %). In the CD3/CD19 group, 91 % had a primary engraftment. After reconditioning, all patients (64/64) were finally engrafted. Patients with CD34-selected or CD133-selected grafts had similar incidences of a GvHD II-IV (3 and 7 %), whereas a GvHD was slightly increased in patients receiving CD3/CD19-depleted cells (27 %). Reconstitution of CD3 (+) T-cells was faster in the CD3/CD19 group than in the CD34 or CD133 group. These preliminary results indicate, that CD3/CD19-selected grafts may be advantageous regarding engraftment and immunoreconstitution. Since effector cell with potential antileukemic activity are cotransfused, such grafts may be suited in particular for patients with insufficient remission. PMID- 16307420 TI - Adenoviral infections after transplantation of positive selected stem cells from haploidentical donors in children: an update. AB - We present updated results of stem cell transplantation with highly purified stem cells from haploidentical parental donors and infection with human adenovirus (HAdV) post stem cell transplantation (SCT). Survival post SCT is primarily determined by relapse, infections and far less by GvHD or other transplant related mortality. During the immune reconstitution the host is at significant risk for severe viral infections. HAdV infection is especially in children an important complication post SCT, with significant morbidity and mortality despite new antiviral treatment strategies. Although control of infection seems to require T-cells, the characterization of HAdV-specific T-cells post SCT has not been introduced in surveillance and treatment decisions. METHODS: Therefore we evaluated the impact of HAdV-infections on the survival between 1995 and 2004 (n = 63) and studied the occurrence of adenovirus-specific T-cells in children with (n = 9) and without (n = 9) HAdV-infection post allogeneic SCT and in healthy donors (n = 53). After stimulation ex-vivo with HAdV-antigen IFN-gamma secreting T-cells were analyzed by flowcytometry and defined as HAdV-specific T-cells. RESULTS: Until day 180 post SCT the cumulative incidence of all lethal viral infections (HAdV n = 5, cytomegalovirus n = 3, herpes simplex virus n = 1) was 16 % for the whole cohort of patients. Cumulative incidence of HAdV-associated mortality was 8.5 %. Cumulative incidence of all lethal viral infections could be now reduced from 16 % to 8 % in conjunction with new surveillance- and therapeutic-strategies. Children with HAdV-associated mortality all had no specific T-cells, although reconstitution of absolute lymphocyte counts exceeded 300/microl within 30 days post transplant. Patients who cleared HAdV infection had normal frequencies of HAdV-specific T-cells until day 200 post SCT. CONCLUSION: In summary adenovirus specific T-cell reconstitution should be monitored in patients after SCT to limit the use of anti viral chemotherapy and help to identify those patients that would benefit from new therapeutic strategies like adoptive transfer of virus specific T-cells. PMID- 16307421 TI - Ex vivo expansion of highly purified NK cells for immunotherapy after haploidentical stem cell transplantation in children. AB - BACKGROUND: Allogeneic natural killer (NK) cells are known to show medium to high cytotoxic activity against HLA-nonidentical leukemia or tumor cells. For a possible benefit of post transplant treatment with NK cells after haploidentical stem cell transplantation (haplo-SCT) we developed a clinical scale procedure for NK cell processing observing Good Manufacturing Practice (GMP). METHODS: Allogeneic donor NK cells were selected from 15 unstimulated leukaphereses using two rounds of immunomagnetic T cell depletion, followed by an NK cell enrichment step. CD56 (+)CD3 (-) NK cells were stimulated and expanded in vitro according to GMP. Quality control of NK cell purity, residual T cells and cytotoxic activity was done by multi-coloured flow cytometric analyses. RESULTS: Purification led to an absolute number of 234-1 237 x 10 (6) CD56 (+)CD3 (-) NK cells from leukapheresis harvests with a median purity of 95 % and a 4 to 6(1/2) log depletion of T cells. After two weeks stimulation with IL-2 a five-fold expansion of NK cells with a T cell contamination below 0.1 % was reached. Median cell viability was 95 % after purification and 99 % after expansion. The IL-2 stimulated NK cells showed a highly increased lytic activity against the MHC-I deficient K562 cells compared to freshly isolated NK cells and a medium cytotoxicity against patients' leukemic cells. CONCLUSIONS: Clinical scale enrichment and activation of allogeneic donor NK cells is feasible. High dose NK cell application may be a new treatment option for pediatric patients with leukemia or solid tumors in case of minimal residual disease or unbalanced chimerism post haplo-SCT as we could show for the first three patients . PMID- 16307422 TI - Adoptive cellular immunotherapy with CD19-specific T cells. AB - BACKGROUND: No effective therapeutic modalities exist for the treatment of relapsed high risk acute lymphoblastic leukemia (ALL). Adoptive cellular immunotherapy by transfusion of polyclonal donor lymphocytes is not always effective and is limited by cellular cross-reactivity with normal tissues, leading to development of clinical graft-versus-host disease (GVHD). METHOD: To develop an immunotherapeutic strategy for targeted elimination of residual leukemic blasts, human T cells were gene-modified to express CD19-specific chimeric receptors. RESULTS: Gene-modified T cells specifically lyse CD19 expressing lymphatic blast cells, however, they show a limited proliferative response to stimulation with CD19. Integration of the signal transduction domain of the costimulatory molecule CD28 enhances the proliferative properties of the gene-modified T cells. CONCLUSIONS: Adoptive transfer of gene-modified virus specific T cells may provide a useful strategy for prevention and early treatment of ALL relapses following allogeneic stem cell transplantation. PMID- 16307423 TI - [Parents-interview on use of complementary and alternative medicine in pediatric oncology in Germany]. AB - BACKGROUND: There are no published data on the use of complementary and alternative medicine (CAM) in pediatric oncology in Germany. PATIENTS/METHOD: All parents of children, who where diagnosed a disease registered in the German Childhood Registry in 2001 were sent a questionnaire to evaluate possible CAM use. RESULTS: 1 602 questionnaires were sent to the families, from whom 1 063 sent the questionnaire back to us. They reported a 35 % CAM-use-rate. The probability to use CAM was positively influenced by preexisting experience with CAM, higher social level and poor prognosis. 71 % of the users discussed the CAM use with a physician. The reason for CAM-use seldom was missing trust in conventional medicine. 89 % of the CAM-users would recommend CAM-use to other parents in a comparable situation. CONCLUSIONS: The reported CAM-use rate of 35 % in pediatric oncology in Germany is as reported in the international literature for other European countries. Essential parts of CAM-advise and -prescription are done by non-professionals, so that there is a need to get competence in CAM in pediatric oncology in Germany. PMID- 16307424 TI - Bleeding and surgery in children with Glanzmann thrombasthenia with and without the use of recombinant factor VII a. AB - BACKGROUND: An inherited deficiency of platelet glycoprotein II b/III a (GP II b/III a), Glanzmann thrombasthenia, can lead to excessive bleeding and require platelet transfusion to secure hemostasis. Antibodies to GP II b/III a or HLA may platelet transfusion render ineffective to stop bleeding or to cover surgery. Recombinant factor VII a has been introduced as therapeutic alternative and has been suggested to be effective. PATIENTS AND AIMS OF THE STUDY: In a retrospective evaluation, bleeding episodes and surgery in six patients treated with antifibrinolytics and with and without the additional use of rFVII a were analysed to achieve informations for treatment indication and efficacy. RESULTS: Nineteen mucosal and subcutaneous bleeding episodes, two dental surgeries and seven joint bleeds occurred. In 11 mild to moderate mucocutaneous bleeds treated without rFVII a, seven stopped within 48 hours, three stopped until the fourth day; one showed recurrence. Three bleeds were treated with rFVII a and responded within 24 hours. One severe bleed treated without rFVII a did not stop until the 8 (th) day after cautery. In 4 severe bleeds treated with rFVII a, one stopped within 24 hours, one showed recurrence, one was treated with platelet transfusion concurrently and one did not respond to rFVII a. Clinical signs persisted in one conservatively treated elbow joint bleed, whereas in two episodes treated with rFVII a, the bleeding responded within 5 and 7 days and in four episodes in at least 4 days. Two dental surgeries showed no recurrence after rFVII a over 18 or 36 hours. CONCLUSIONS: In severe mucocutaneous bleeding episodes or joint bleeding rFVII a is of some benefit whereas in surgeries like teeth extraction, prophylactically administered rFVII a seems effective to avoid bleeding. In mild to moderate mucocutaneous bleeding events, antifibrinolytics and local measures were sufficient in most cases and the additional use of rFVII a does not seem to be necessary. Further information is needed to elaborate clear indications for the rational use of rFVII a in bleeding episodes in patients with Glanzmann thrombasthenia compared to standardized baseline treatment. This information may generate a prospective multicenter study to provide clear advice with respect to bleeding site, severity and duration. PMID- 16307425 TI - [Duration of storage of autologous hematopoietic stem cell products for children and adolescents with malignant diseases]. PMID- 16307426 TI - Fourier Transform Infrared Imaging of focal lesions in human osteoarthritic cartilage. AB - Fourier Transform Infrared Imaging (FTIRI) is a new method for quantitatively assessing the spatial-chemical composition of complex materials. This technique has been applied to examine the feasibility of measuring changes in the composition and distribution of collagen and proteoglycan macromolecules in human osteoarthritic cartilage. Human cartilage was acquired post-operatively from total joint replacement patients. Samples were taken at the site of a focal lesion, adjacent to the lesion, and from relatively healthy cartilage away from the lesion. Sections were prepared for FTIRI and histochemical grading. FTIRI spectral images were acquired for the superficial, intermediate, and deep layers for each sample. Euclidean distance mapping and quantitative partial least squares analysis (PLS) were performed using reference spectra for type-II collagen and chondroitin 6-sulphate (CS6). FTIRI results were correlated to the histology-based Mankin scoring system. PLS analysis found relatively low relative concentrations of collagen (38 +/- 10%) and proteoglycan (22 +/- 9%) in osteoarthritic cartilage. Focal lesions were generally found to contain less CS6 compared to cartilage tissue adjacent to the lesion. Loss of proteoglycan content was well correlated to histological Mankin scores (r=0.69, p<0.0008). The evaluation of biological tissues with FTIRI can provide unique quantitative information on how disease can affect biochemical distribution and composition. This study has demonstrated that FTIRI is useful in quantitatively assessing pathology-related changes in the composition and distribution of primary macromolecular components of human osteoarthritic cartilage. PMID- 16307427 TI - Recent advances in the application of capillary electromigration methods for food analysis. AB - This article reviews the latest developments in the application of capillary electromigration methods for the analysis of foods and food components. Nowadays, methods based on CE techniques are becoming widely used in food analytical and research laboratories. This review covers the application of CE to analyze amino acids, biogenic amines, peptides, proteins, DNAs, carbohydrates, phenols, polyphenols, pigments, toxins, pesticides, vitamins, additives, small organic and inorganic ions, chiral compounds, and other compounds in foods, as well as to investigate food interactions and food processing. The use of microchips as well as other foreseen trends in CE analysis of foods is discussed. Papers that were published during the period June 2002-June 2005 are included following the previous review by Frazier and Papadopoulou (Electrophoresis 2003, 24, 4095 4105). PMID- 16307428 TI - Advances in capillary electrophoretically mediated microanalysis: an update. AB - This review, as a continuation of an earlier report, gives an overview of recent developments, over the period from 2003 until now, in the use of capillary electrophoretic techniques for the in-line study of enzymatic reactions, derivatization, and chemical reactions. The article is divided into two parts: (i) in-line enzymatic reactions and (ii) in-line derivatization and chemical reactions. The first part introduces electrophoretically mediated microanalysis (EMMA) and discusses and illustrates the different modes of EMMA. A literature overview on enzymatic reactions is provided. The second part starts with an introduction of the procedures and the nomenclature used in the area of in-line derivatization and chemical reactions based on EMMA. Reported derivatization and chemical reaction procedures are discussed and summarized. PMID- 16307429 TI - Recent developments in capillary electrophoresis and capillary electrochromatography of peptides. AB - The article gives a comprehensive review on the recent developments in the applications of high-performance capillary electromigration methods, zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography, to analysis, preparation, and physicochemical characterization of peptides. The article presents new approaches to the theoretical description and experimental verification of electromigration behavior of peptides, covers the methodological aspects of capillary electroseparations of peptides, such as rational selection of separation conditions, sample preparation, suppression of peptide adsorption, new developments in individual separation modes, and new designs of detection systems. Several types of applications of capillary electromigration methods to peptide analysis are presented: conventional qualitative and quantitative analysis, purity control, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Some examples of micropreparative peptide separations are given and capabilities of capillary electromigration techniques to provide important physicochemical characteristics of peptides are demonstrated. PMID- 16307430 TI - Capillary electrophoresis of proteins 2003-2005. AB - This review article with 304 references describes recent developments in CE of proteins, and covers the two years since the previous review (Hutterer, K., Dolnik, V., Electrophoresis 2003, 24, 3998-4012) through Spring 2005. It covers topics related to CE of proteins, including modeling of the electrophoretic migration of proteins, sample pretreatment, wall coatings, improving separation, various forms of detection, special electrophoretic techniques such as affinity CE, CIEF, and applications of CE to the analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals, and recombinant protein preparations. PMID- 16307431 TI - Analysis of explosives via microchip electrophoresis and conventional capillary electrophoresis: a review. AB - The upsurge in terrorist activity has generated tremendous demand for innovative tools capable of detecting major industrial, military, and home-made (improvised) explosives. Fast, sensitive, and reliable detection of explosives in the field is a very important issue in nowadays. CE, especially in its miniaturized format (lab-on-a-chip), offers great possibilities to create portable, field deployable, rapidly responding, and potentially disposable devices, allowing security forces to make the important decisions regarding the safety of civilians. This article overviews the microchip and conventional capillary electrophoretic techniques for analysis of a wide variety of explosive compounds and mixtures. PMID- 16307432 TI - Interaction of polyanions with basic proteins, 2(a) : influence of complexing polyanions on the thermo-aggregation of oligomeric enzymes. AB - The ability of synthetic polyanions to suppress thermo-aggregation of the oligomeric enzymes (glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase, and aspartate aminotransferase) has been established. The ability of the polyanions to reduce the thermo-aggregation increased in the order poly(methacrylic acid) < poly(acrylic acid) < sodium poly(styrene sulphonate), which agreed well with the increase, in the same order, of the charge density of the chains. The lengthening of the chains, as well as the rise in their relative content, resulted in an increase of the ability to reduce thermo-aggregation, mentioned above. Complete prevention of the enzyme aggregation was achieved when highly charged polyanions of a relatively high degree of polymerization were used in a concentration sufficient to solubilize the protein. Complexing with the polyanions prevented thermo-aggregation of the enzymes, but not their thermo denaturation. The adverse effect of the complexing polyanions on the catalytic activity was reduced by the addition of a synthetic polycation, which resulted in a significant reactivation (up to 40%) of the enzyme. The possibility of preventing the thermo-aggregation of enzyme molecules and then partly restoring the enzyme activity, appears to be of particular interest when studying the aggregation mechanism of proteins that are prone to form the amyloid structures responsible for the development of neurodegenerative diseases like Alzheimer's disease, bovine spongiform encephalopathy and Huntington disease. This finding can also be considered as an important step in the creation of artificial chaperones. PMID- 16307433 TI - Multilayer microcapsules as anti-cancer drug delivery vehicle: deposition, Sustained release, and in vitro bioactivity. AB - A drug delivery system based on spontaneous deposition of soluble, low-molecular weight therapeutic agents has been developed for the purpose of sustaining drug release. Layer-by-layer assembly of oppositely charged polyelectrolytes onto melamine formaldehyde (MF) colloidal particles, followed by removal of the cores at low pH has yielded intact hollow microcapsules having the ability to induce deposition of various water-soluble substances. Dynamic observation by confocal laser scanning microscopy provided direct evidence of such deposition. Dependence of loading rate on molecular weight was investigated. Efficient loading of an anti-cancer drug, daunorubicin (DNR), was confirmed by transmission electron microscopy (TEM). Its release was quantified by fluorometry. The results indicated that loading, and subsequent release, could be tuned by factors such as feeding concentrations, temperature, and salt concentrations. The intrinsic mechanism of loading and release was discussed taking into account the interaction between the drugs and the poly(styrene sulfonate)/MF complex existing in the hollow capsules. With culture of the HL-60 cell line, a kind of human leukemia cell, the presence of DNR-loaded capsules was seen to steadily decrease the cyto-viability. Fluorescence intensity averaged from inside the circles as a function of incubation time. PMID- 16307434 TI - Biodegradable hyaluronic acid/n-carboxyethyl chitosan/protein ternary complexes as implantable carriers for controlled protein release. AB - An ampholytic N-carboxyethyl chitosan (CEC), with various isoelectric points (IPs), was synthesized by grafting acrylic acid on chitosan utilizing Michael's reaction. Compared to native chitosan, CEC has enhanced water solubility and dramatically accelerated enzymatic degradation; the rate of degradation is proportional to the degree of substitution (DS). The results from turbidimetric titration and fluorescence studies revealed that CEC formed complexes with either hyaluronic acid (HA) or bovine serum albumin (BSA) within a certain pH range. The HA/CEC/BSA ternary complexes could be prepared by colloid titration with quantitative yield and BSA entrapment. The rate of BSA release from the complexes was affected by pH, ionic strength, DS of CEC, and the molecular weight (MW) of HA. The endurance of BSA release from the complexes could be extended up to 20 d by formulating them with high-MW HA and CEC with low DS.BSA release profiles from HA/CEC-2/BSA complexes. PMID- 16307436 TI - Potential bias in Generalized Estimating Equations linkage methods under incomplete information. AB - The mean identity-by-descent (IBD) specification used in the Generalized Estimating Equations (GEE) methodology for linkage is only valid, strictly speaking, under the assumption of fully polymorphic markers. In practice, markers often provide only partial IBD information, which can potentially result in inconsistency of the locus location and gene effect estimates obtained by the GEE method. Using both simulations and theory, we identify some realistic conditions about marker information under which the validity of the GEE linkage methods may be arguable. Namely, researchers should not trust the GEE parameters' estimates and their associated confidence intervals in areas of the genome where IBD information is sparse or when this information changes abruptly. We show that properly standardized statistics based on IBD sharing provide a valid alternative. PMID- 16307437 TI - Overexpression of ErbB2 and ErbB3 receptors in Schwann cells of patients with Charcot-Marie-tooth disease type 1A. AB - Axon-derived neuregulins (NRGs) are a family of growth factors whose binding to ErbB tyrosine kinase receptors promotes the maturation, proliferation and survival of Schwann cells (SCs). Correct NRG/ErbB signaling is essential for the homeostasis of axonal-glial complexes and seems to play a role in peripheral nerve repair. The potential involvement of ErbB receptors in human peripheral neuropathies has not been clarified. Therefore, we assessed the immunoreactivity for EGFR (ErbB1), ErbB2, and ErbB3 in nerve biopsies from patients with different forms of Charcot-Marie-Tooth disease, type 1, (CMT1), as compared to others with inflammatory neuropathies and controls. The most notable changes consisted in the overexpression of ErbB2 and ErbB3 by SCs of nerves from CMT1A patients. These findings are consistent with an impairment of SC differentiation and expand the molecular phenotype of CMT1A. The upregulation of these receptors may play a role in the inhibition of myelination or in the promotion of recurrent demyelination and axonal damage. PMID- 16307438 TI - Coactivation of the elbow antagonist muscles is not affected by the speed of movement in isokinetic exercise. AB - Since muscle coactivation increases the stiffness and stability of a joint, greater coactivation is likely during faster than slower movements. Very few studies, though, have been conducted to verify this hypothesis. Moreover, a large number of studies have examined coactivation of muscles surrounding the knee joint whereas there are few reports on the elbow joint. The aim of this study was therefore to compare the antagonist activation of the elbow flexors and extensors during isokinetic concentric exercises and to investigate the influence of angular velocity on their activation. Twelve men participated in the study. The surface electromyographic signals (sEMG) were recorded from the biceps brachii (BB) and triceps brachii (TB) muscles during three maximal voluntary isometric contractions (MVC) of elbow flexors and extensors and a set of three maximal elbow flexions and extensions each at 15 degrees, 30 degrees , 60 degrees, 120 degrees, 180 degrees, and 240 degrees.s(-1). Normalized root mean square (RMS) of sEMG was calculated during the isokinetic phase of movement as an index of sEMG amplitude. During elbow flexion, the antagonist activation of BB averaged 16.2% lower than TB, and this difference was statistically significant at all angular velocities. The normalized RMS values ranged from 26.0% +/- 19.0 at MVC to 37.8% +/- 13.9 at 240 degrees.s(-1) for antagonist TB activation, and from 5.7% +/- 5.2 at MVC to 18.9% +/- 8.6 at 240 degrees.s(-1) for antagonist BB activation. No influence of angular velocity on agonist and antagonist activity was found. Moreover, flexion and extension torques were both strongly affected by the amount of antagonist activation. The functional specialization of the two muscle groups could be responsible for the different levels of antagonist activation. The frequent use of BB, which is not assisted by gravity during daily activities, could lead to reduced coactivation due to a better functioning of the control system based upon reciprocal innervation. These findings may have significant implications in the design of rehabilitation programs directed to the elbow joint. PMID- 16307439 TI - Motor evoked potentials in a mouse model of chronic multiple sclerosis. AB - We tested cortical motor evoked potentials (cMEPs) as a quantitative marker for in vivo monitoring of corticospinal tract damage in a murine multiple sclerosis model (experimental autoimmune encephalomyelitis, EAE). The cMEPs, previously standardized in naive C57BL/6 developing and adult mice, were studied longitudinally in adult EAE mice. Central conduction times (CCTs) increased significantly shortly before the earliest clinical signs developed (10 days postimmunization, dpi), with peak delay in acute EAE (20-40 dpi). In clinically stable disease (80 dpi), CCTs did not increase further, but cMEP amplitude declined progressively, with complete loss in >80% of mice at 120 dpi. Increase in CCT correlated with presence of inflammatory infiltrates and demyelination in acute EAE, whereas small or absent cMEPs were associated with continuing axonal damage in clinically-stabilized disease and beyond (>80 dpi). These results demonstrate that cMEPs are a useful method for monitoring corticospinal tract function in chronic-progressive EAE, and provide insight into the pathological substrate of the condition. PMID- 16307440 TI - Related timing for peripheral and central plasticity in hypoglossal-facial nerve anastomosis. AB - The aim of this work was to determine the role of peripheral facial muscle reinnervation in the central reorganization of the blink reflex (BR) after hypoglossal-facial anastomosis (HFA). An electrophysiological study was performed on seven patients who underwent HFA after facial nerve transection during surgery for acoustic neuroma. HFA was performed within 15 days after surgery in five patients (group 1) and later for the two others (group 2). We studied the motor responses (MR) and the BR evoked on the affected side, before and over 3 years after the HFA. The MR appeared by the third month for the first group, and by the sixth and twelfth for the second group. After 36 months, the amplitude of MR was significantly higher than its control value, showing hyperinnervation of the facial muscles. Study of the BR evoked only an R1-type blink response that was observed 4 and 6 months after the MR for groups 1 and 2, respectively. This central reorganization appeared closely correlated with muscle reinnervation and its related timing. The occurrence of peripheral nerve-muscle contacts seems to be a necessary condition for reorganization of the trigemino-hypoglossal-facial reflex. PMID- 16307441 TI - The electromyographic signal as a presymptomatic indicator of organophosphates in the body. AB - Organophosphate (OP) compounds are present in household and agricultural pesticides as well as in nerve agents. The toxic effects of these chemicals result from their anticholinesterase activity, which disrupts nerve junctions and parasympathetic effector sites, leading to a variety of symptoms and possible death. When the anticholinesterase agents in OP compounds reach the neuromuscular junction, they cause a disruption in the firing of muscle fiber action potentials. This effect has the potential of altering the time course of the electromyographic (EMG) signal detected by surface electrodes. We investigated the association between OP compound dose, surface EMG changes, and overt signs of OP toxicity. Daily doses of 10-15 microg/kg of diisopropylfluorophosphate (DFP) were injected into the calf muscle of four rhesus monkeys while surface EMG signals were recorded from two thigh muscles bilaterally. With increasing number of doses, the EMG signal presented an increasing number of time gaps. The presence of the gaps was evident prior to any overt symptoms of cholinesterase toxicity. These findings can lead to the development of noninvasive technology for indicating the presence of OP compounds in muscle tissue prior to clinical abnormalities. PMID- 16307442 TI - Which patients need referral for anal sphincter electromyography? AB - Needle electromyography (EMG) of lower sacral myotomes is useful in certain patients with urinary, bowel, or sexual dysfunction. The aim of the study was to identify the clinical profile of patients who require such testing. Medical records were retrospectively reviewed, and findings evaluated using bivariate and multivariate statistics. A neuropathic condition affecting the lower sacral segments was diagnosed by quantitative concentric needle EMG of the external anal sphincter (EAS) muscle. Neuropathic changes in the EAS were found in 85 (44%) of 193 patients studied. On ordinal logistic regression analysis, bladder-emptying difficulties and perineal sensory loss were significantly related to the presence of a neuropathic EMG. No guidelines for referral for anal sphincter EMG could be defined that would include all patients with neuropathic abnormalities in the lower sacral segments. However, patients with bladder-emptying difficulties and perineal sensory loss seem to be the most suitable candidates for testing. PMID- 16307443 TI - Dartos reflex: a sympathetically mediated scrotal reflex. AB - The dartos muscle is a sympathetically innervated dermal muscle layer within the scrotum, distinct from the somatically innervated cremasteric muscle. We electrophysiologically demonstrate the presence of a dartos reflex (DR), which can be used to evaluate the thoracolumbar sympathetic and genitofemoral nerve pathways. In 20 healthy men, we evoked the DR by cutaneous stimulation of the thigh and recorded the resultant scrotal skin contraction. We recorded hand, foot, and perineal sympathetic skin responses (SSRs) as controls. The DR was reliable and reproducible, as were the SSRs. The mean left DR latency was 4.8 s (SD, 2.7 s) and right DR latency was 5.4 s (SD, 3 .4), both of which were longer than the mean hand, foot, and perineal SSRs (P < 0.05). An intact reflex arc reflects the integrity of the afferent and efferent branches of the genitofemoral nerve (T12-L2). The DR test can also be used to assess scrotal autonomic innervation. Abnormalities of dartos innervation may impact testis thermoregulation and spermatogenesis. PMID- 16307444 TI - c-Jun N-terminal kinase inhibition and alpha-tocopherol protect midbrain dopaminergic neurons from interferon-gamma/lipopolysaccharide-induced injury without affecting nitric oxide production. AB - Interferon-gamma (IFN-gamma)/lipopolysaccharide (LPS) induces delayed dopaminergic neuron loss in midbrain slice cultures, because of nitric oxide production resulting from p38 mitogen-activated protein kinase (p38 MAPK) dependent induction of inducible nitric oxide synthase (iNOS). In this study, we show that inhibition of c-Jun N-terminal kinase (JNK), but not of extracellular signal-regulated kinase, protects dopaminergic neurons from IFN-gamma/LPS-induced degeneration. In contrast to a p38 MAPK inhibitor, SB203580, however, a JNK inhibitor, anthra[1,9-cd]pyrazol-6(2H)-one (SP600125), did not suppress IFN gamma/LPS-induced iNOS expression and nitric oxide production. Involvement of NADPH oxidase-derived superoxide production in dopaminergic neurodegeneration was not obvious, in that superoxide dismutase/catalase or manganese 3-methoxy-N,N' bis(salicylidene)ethylenediamine chloride (EUK-134), a superoxide dismutase/catalase mimetic, did not afford neuroprotection. Moreover, the NADPH oxidase inhibitors apocynin and diphenylene iodonium were protective against IFN gamma/LPS cytotoxicity only at concentrations that suppressed nitric oxide production. Notably, alpha-tocopherol effectively prevented IFN-gamma/LPS-induced dopaminergic neuron degeneration, without affecting iNOS induction and nitric oxide production. These results underscore the neuroprotective potential of JNK inhibitor and alpha-tocopherol, in the sense that both agents could rescue dopaminergic neurons under inflammatory conditions associated with robust increases in nitric oxide production. PMID- 16307445 TI - Neural progenitor number is regulated by nuclear factor-kappaB p65 and p50 subunit-dependent proliferation rather than cell survival. AB - The number of cells generated by a proliferating stem or precursor cell can be influenced both by proliferation and by the degree of cell death/survival of the progeny generated. In this study, the extent to which cell survival controls progenitor number was examined by comparing the growth characteristics of neurosphere cultures derived from mice lacking genes for the death-inducing Bcl-2 homologue Hara Kiri (Hrk), apoptosis-associated protein 1 (Apaf1), or the prosurvival nuclear factor-kappaB (NFkappaB) subunits p65, p50, or c-rel. We found no evidence that Hrk or Apaf1, and by inference the mitochondrial cell death pathway, are involved in regulating the number of neurosphere-derived progeny. However, we identified the p65p50 NFkappaB dimer as being required for the normal growth and expansion of neurosphere cultures. Genetic loss of both p65 and p50 NFkappaB subunits resulted in a reduced number of progeny but an increased proportion of neurons. No effect on cell survival was observed. This suggests that the number and fate of neural progenitor cells are more strongly regulated by cell cycle control than survival. PMID- 16307446 TI - Ischemic preconditioning is mediated by erythropoietin through PI-3 kinase signaling in an animal model of transient ischemic attack. AB - Ischemic preconditioning (IP) protects the brain from subsequent, prolonged, and lethal ischemia in experimental studies. Erythropoietin (EPO) participates in the brain's intrinsic response to injury and may play a role in preconditioning. By using a middle cerebral artery occlusion (MCAo) model of transient ischemic attack (TIA), we sought to determine whether EPO is required for IP in the protective response against focal ischemic stroke. Rats underwent three 10-min MCA occlusions or sham surgery. Three days later, animals underwent 2 hr of MCAo and 22 hr of reperfusion. Experimental TIAs reduced infarct volumes by 55% (P < 0.05), inhibited DNA fragmentation, and improved neurological outcome by 50% (P < 0.05) after ischemic stroke. EPO and its receptor were up-regulated by IP in the ipsilateral hemisphere by 24 hr after IP, before ischemic stroke and soluble EPO receptor attenuated neuroprotection by IP (88% reduction, P < 0.05). Pretreatment with the PI-3 kinase inhibitor wortmannin abolished the protective effect of IP against ischemic injury (P < 0.05). IP may be mediated in part by EPO through a PI-3 kinase pathway. PMID- 16307447 TI - Effects of cochlear ablation on muscarinic acetylcholine receptor binding in the rat cochlear nucleus. AB - Cholinergic synapses in the cochlear nucleus (CN) have been reported to modulate spontaneous activity via muscarinic acetylcholine receptors. In this study, muscarinic receptor binding was measured as specific binding of 1-[N-methyl (3)H]scopolamine in CN regions of control rats and 7 days, 1 month, and 2 months after unilateral cochlear ablation. In control rats, the strongest binding was found in granular regions, followed in order by fusiform soma, molecular, and deep layers of the dorsal cochlear nucleus (DCN), with much lower binding in the anteroventral CN (AVCN) and posteroventral CN (PVCN). After unilateral cochlear ablation, binding in the AVCN, PVCN, and their associated granular regions on the lesion side became progressively greater than on the control side through 2 months after lesion. A significant asymmetry, with binding higher on the lesion side, was also found in the DCN fusiform soma layer at 7 days, and there and in the DCN deep layer at 1 and 2 months after lesion. There was also evidence of increased binding on the control side in most CN regions. By contrast, binding in the ipsilateral facial nucleus decreased, compared with the control side, by 7 days after the lesion and showed some recovery toward symmetry by 2 months after lesion, and there was no evidence for contralateral changes. These muscarinic receptor binding changes reflect receptor plasticity after loss of auditory nerve innervation. Such plasticity may underlie some of the central auditory functional changes that occur following peripheral lesions, such as tinnitus and hyperacusis. PMID- 16307448 TI - Involvement of alpha1beta1 integrin in insulin-like growth factor-1-mediated protection of PC12 neuronal processes from tumor necrosis factor-alpha-induced injury. AB - Insulin-like growth factor 1 receptor (IGF-1R) supports neuronal survival against a wide variety of insults. This includes tumor necrosis factor-alpha (TNFalpha) mediated neuronal damage, which represents one of the factors suspected to play a role in HIV-associated dementia (HAD). PC12 neurons engineered to express human IGF-1R (PC12/IGF-1R) maintain neuronal processes on collagen IV for several weeks. However, prolonged treatment with TNFalpha caused degeneration of neuronal processes, with no apparent signs of apoptosis. In this process, TNFalpha did not affect IGF-1-mediated phosphorylation of IRS-1, IRS-2, Akt, or Erks. In addition, PC12/IGF-1R cells were found to express predominantly alpha1beta1 integrin, which has high affinity to collagen IV. The treatment of PC12/IGF-1R neurons with a specific alpha1beta1 integrin inhibitor, obtustatin, also caused loss of neuronal processes, accompanied by a quick cell detachment and extensive apoptosis. In the presence of IGF-1, both TNFalpha-induced and obtustatin-induced degeneration of neuronal processes were effectively inhibited. Furthermore, TNFalpha-mediated neuronal degeneration correlated with decreased attachment of PC12/IGF-1R cells to collagen IV and with a reduced level of alpha1beta1 integrin, consistent with a role for this surface protein in the maintenance of neuronal processes. Thus the neuroprotective effects of IGF-1 are not restricted to its antiapoptotic properties but also involve an additional neuroprotective mechanism, by which IGF 1 counteracts the negative effect of TNFalpha on alpha1beta1 integrin-mediated attachment to collagen IV. PMID- 16307449 TI - Nicotine prevents stress-induced enhancement of long-term depression in hippocampal area CA1: electrophysiological and molecular studies. AB - Nicotine treatment prevents chronic psychosocial stress-induced impairment of hippocampus-dependent spatial memory and long-term potentiation (LTP). In this study, we investigated the effect of chronic nicotine treatment on stress-induced enhancement of long-term depression (LTD). After paired-pulse stimulation, LTD was evoked in area CA1 of anesthetized control, stressed, nicotine-treated, and nicotine-treated stressed rats. In stressed rats, a significantly greater LTD magnitude was seen than in control rats. Stress also facilitated the induction of LTD. Nicotine treatment of stressed rats prevented stress-induced enhancement and facilitation of LTD. For chronically stressed rats, we previously reported marked decreases in the basal levels of brain-derived neurotrophic factor (BDNF), CaMKII, P-CaMKII, and calmodulin as well as a significant increase in calcineurin basal levels. Herein, Western blot analysis conducted 1 hr after induction of LTD by paired-pulse stimulation showed that the levels of calcineurin and P-CaMKII were increased in the stressed group compared with the other groups and were normalized by chronic nicotine treatment. Additionally, after paired-pulse stimulation, the levels of total CaMKII were increased in all groups with no change in the levels of BDNF and calmodulin. Therefore, the increase in the levels of calcineurin and P-CaMKII during expression of LTD in area CA1 may explain the enhanced magnitude of LTD in chronically stressed rats. PMID- 16307450 TI - JIL-1 kinase, a member of the male-specific lethal (MSL) complex, is necessary for proper dosage compensation of eye pigmentation in Drosophila. AB - The upregulation of the JIL-1 kinase on the male X chromosome and its association with the male-specific lethal (MSL) complex suggest that JIL-1 may play a role in regulating dosage compensation. To directly test this hypothesis we measured eye pigment levels of mutants in the X-linked white gene in an allelic series of JIL 1 hypomorphic mutants. We show that dosage compensation of w(a) alleles that normally do exhibit dosage compensation was severely impaired in the JIL-1 mutant backgrounds. As a control we also examined a hypomorphic white allele w(e) that fails to dosage compensate in males due to a pogo element insertion. In this case the relative pigment level measured in males as compared to females remained approximately the same even in the most severe JIL-1 hypomorphic background. These results indicate that proper dosage compensation of eye pigment levels in males controlled by X-linked white alleles requires normal JIL-1 function. PMID- 16307451 TI - Biowaiver monographs for immediate release solid oral dosage forms: acetaminophen (paracetamol). AB - Literature data are reviewed on the properties of acetaminophen (paracetamol) related to the biopharmaceutics classification system (BCS). According to the current BCS criteria, acetaminophen is BCS Class III compound. Differences in composition seldom, if ever, have an effect on the extent of absorption. However, some studies show differences in rate of absorption between brands and formulations. In particular, sodium bicarbonate, present in some drug products, was reported to give an increase in the rate of absorption, probably caused by an effect on gastric emptying. In view of Marketing Authorizations (MAs) given in a number of countries to acetaminophen drug products with rapid onset of action, it is concluded that differences in rate of absorption were considered therapeutically not relevant by the Health Authorities. Moreover, in view of its therapeutic use, its wide therapeutic index and its uncomplicated pharmacokinetic properties, in vitro dissolution data collected according to the relevant Guidances can be safely used for declaring bioequivalence (BE) of two acetaminophen formulations. Therefore, accepting a biowaiver for immediate release (IR) acetaminophen solid oral drug products is considered scientifically justified, if the test product contains only those excipients reported in this paper in their usual amounts and the test product is rapidly dissolving, as well as the test product fulfils the criterion of similarity of dissolution profiles to the reference product. PMID- 16307452 TI - Application of an exact mathematical model and the steady-state approximation to the kinetics of the reaction of cysteine and hydrogen peroxide in aqueous solution: a reply to the Ashby and Nagy commentary. PMID- 16307453 TI - Transport of drugs in the kidney by the human organic cation transporter, OCT2 and its genetic variants. AB - The human organic cation transporter 2 (OCT2, SLC22A2) is a multispecific transporter of organic cations, including many clinically used drugs. OCT2 is primarily responsible for the uptake of organic cations across the basolateral membrane of renal tubular epithelial cells and is considered a major transporter in the active secretion of organic cations in the kidney. Uptake of organic cations by OCT2 is driven by the inside-negative membrane potential and is pH sensitive. Regulation of OCT2 at the transcriptional level by steroid hormones and at the protein level by various protein kinases has been described. Several human genetic variants in the coding region of OCT2 have been identified and functionally characterized, including both polymorphic and rare variants. A variety of structurally diverse compounds have been shown to interact with OCT2, including endogenous compounds, drugs, and dietary supplements. PMID- 16307454 TI - A rabbit model for sublingual drug delivery: comparison with human pharmacokinetic studies of propranolol, verapamil and captopril. AB - A rabbit model for investigating sublingual drug absorption was established yielding results consistent with clinical data reported in the literature. Using propranolol as a model compound the effect of formulation and dosing variables was explored as a means to characterize the limiting parameters of this model. In addition, verapamil and captopril were selected as reference compounds to compare this model to sublingual absorption in humans. Rabbits were dosed sublingually and systemic absorption was measured over time. Sublingual absorption of propranolol was dependent on dosing solution pH and volume. Intra-oral spray device did not affect the overall exposure compared to instillation using a syringe. Despite species and dosing regimen differences the relative bioavailabilities of propranolol and verapamil were very similar in rabbits and humans. In contrast, captopril absorption from the sublingual cavity of rabbits was low and did not agree with that observed in man. Here we report a sublingual rabbit model of drug delivery and its potential utility in preclinical development of intra-oral dosage forms. PMID- 16307455 TI - Effect of different surfactants in biorelevant medium on the secretion of a lipophilic compound in lipoproteins using Caco-2 cell culture. AB - The impact of a pharmaceutical relevant metabolizable, ionic surfactant or two synthetic, nonionic surfactants on the absorption and lipoprotein incorporation of a lipophilic drug, retinol, was studied in the Caco-2 cell culture. Filter grown monolayers of Caco-2 cells were incubated for 20 h with (3)H-retinol and (14)C-oleic acid and with increasing concentrations of lyso-phosphatidylcholine (lyso-PC), Cremophor RH40, or Tween 80. The concentration of (3)H-retinol and (14)C-lipid was measured in the apical, intracellular, and basolateral compartments. The basolateral medium was ultracentrifugated into different lipoprotein classes and their (3)H-retinol and (14)C-lipid concentrations were determined. The cells incubated with lyso-PC and Tween 80 increased the incorporation of (3)H-retinol and (14)C-lipid into chylomicrons and very low density lipoproteins (VLDL). The explored surfactants impacted the incorporation of (3)H-retinol and (14)C-lipid in chylomicrons and VLDL in a concentration dependent manner. As these surfactants interfere with the intestinal lipoprotein secretion, inclusion of high concentrations of the surfactants in lipid-based formulations of poorly aqueous soluble drugs might impact the degree of intestinal lymphatic transport of the drug after oral administration. PMID- 16307456 TI - Investigating the hydrate conversion propensity of different etoricoxib lots. AB - The physical stability of bulk active pharmaceutical ingredients (API) is of significant scientific and regulatory concern. Carrying out physical stability testing on lots with varying rates of hydrate conversion can potentially lead to erroneous conclusions if these rate differences remain unknown and unstudied. The lot dependency of etoricoxib's rate of hemihydrate conversion was investigated and a quick discriminatory technique was developed to qualitatively assess relatively slow to rapidly converting lots. This novel technique was also used to screen potential parameters affecting the hydrate conversion rate such as particle size/surface area, amorphous content, and initial hemihydrate content. Based on qualitative X-ray powder diffraction (XRPD) and quantitative Raman data, significant effects on the rate of hydration were observed with the addition of small amounts of amorphous etoricoxib. Furthermore, it was found that the presence of hemihydrate also increased the rate of conversion by seeding anhydrous etoricoxib. This suggests that the initial presence of the hydrate form can cooperatively accelerate conversion. A better understanding of the factors affecting hydrate conversion rates resulted in the appropriate selection of storage conditions for both the bulk API and the formulated product. PMID- 16307457 TI - Assay of naproxen by high-performance liquid chromatography and identification of its photoproducts by LC-ESI MS. AB - A rapid, accurate and reliable reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of naproxen and its photodegradation products in methanol was developed and validated. An Inertsil 5 ODS-3V column (5 microm, C18, 250 x 4.6 mm i.d.) was used with a mobile phase of acetonitrile-methanol-1% HOAc in H2O (40:20:40, v/v/v). UV detection was set at 230 nm. The developed method satisfies system suitability criteria, peak integrity and resolution for the parent drug and its photoproducts. The intraday and interday standard deviations of five replicate determinations for five consecutive days at the working concentrations of 5.0, 10, 25, 50, and 100 microm were 0.23-0.98 with coefficients of variance (CVs) of between 0.96 and 4.56% for the former, and 0.14-1.15 with CVs of between 1.13 and 3.82% for the latter. The percentage recoveries were determined to be 98.34, 99.19, 100.18, 102.97 and 99.81%, respectively, at the five concentrations between 5.0 and 100 microm. The limit of quantitation of naproxen was determined to be 0.29 microg/mL, while the detection limit was 64 ng/mL. Four major photoproducts were observed from the HPLC chromatogram using a Panchum PR-2000 reactor which equipped with 8 W x 16 low-pressure quartz mercury lamps as the light source for irradiation of a naproxen sample in methanol. The structures of the photoproducts were confirmed by LC-ESI MS. PMID- 16307458 TI - Nodal and splenic marginal zone B cell lymphomas. AB - Splenic marginal zone lymphoma (SMZL) and nodal marginal zone lymphoma (NMZL) are newly defined, separate clinicopathological entities. Both are rare lymphoma types, with low reproducibility in the diagnosis, although a conjunction of molecular and clinical studies seems to be now facilitating a more accurate diagnosis and understanding of the neoplastic process. SMZL is a disease involving the spleen, bone marrow and peripheral blood since the initial manifestations of the disease. The diagnosis has been until very recently based on the pathological study of the spleen with the conjunction of the clinical features, although the integration of the morphology in bone marrow and peripheral blood with the immunophenotype and molecular characteristics of the tumour makes a more accurate diagnosis now possible. The most frequent molecular alteration found in SMZL is allelic loss at the 7q chromosomal region. SMZL is an indolent lymphoma, although there is small subset of patients in which it follows an aggressive course. Molecular studies of SMZL are starting to reveal new diagnostic and prognostic markers, and to identify new potentially useful therapeutic targets. Nodal marginal zone lymphoma is a B-cell neoplasm originated in the lymph node, whose histology resembles the nodal infiltration by MALT- or Splenic-type marginal zone lymphoma, in the absence of clinical evidence of extranodal or spleen disease. The lack of characteristic phenotypic or molecular diagnostic findings is still hampering the reproducibility of this diagnosis. Here we review the main morphological and immunophenotypical markers, discussing the differential with other overlapping entities, singularly follicular lymphoma. Specific therapeutic protocols and prognostic factors are required to more precisely define this tumour. PMID- 16307459 TI - Water oligomers in crystal hydrates--what's news and what isn't? PMID- 16307460 TI - Highly efficient reagents for Br/Mg exchange. PMID- 16307461 TI - Elucidation of the electrochemical activation of water over Pd by first principles. PMID- 16307462 TI - Arsenic fatty acids are human urinary metabolites of arsenolipids present in cod liver. PMID- 16307463 TI - Replacement of isobutyl by trifluoromethyl in pepstatin A selectively affects inhibition of aspartic proteinases. AB - Two bis-trifluoromethyl pepstatin A analogues, carboxylic acid 1 and its methyl ester 2, have been synthesised in order to probe the properties and size of the trifluoromethyl (Tfm) group and compare it to the "bigger" isobutyl that is present in pepstatin A. The results demonstrate that Tfm can effectively replace the isobutyl chain as far as inhibitory activity against plasmepsin II (PM II), an aspartic proteinase from Plasmodium falciparum, is concerned. On the other hand, replacement of isobutyl by Tfm selectively affected activity against other aspartic proteinases tested. Two lines of evidence led to these conclusions. Firstly, compounds 1 and 2 retained single-digit nanomolar inhibitory activity against PM II, but were markedly less active against PM IV, cathepsin D and cathepsin E. Secondly, the X-ray crystal structures of the three complexes of PM II with 1, 2 and pepstatin A were obtained at 2.8, 2.4 and 1.7 A resolution, respectively. High overall similarity among the three complexes indicated that the central Tfm was well accommodated in the lipophilic S1 pocket of PM II, where it was involved in tight hydrophobic contacts. The interaction of PM II with Phe111 appeared to be crucial. Comparison of the crystal structures presented here, with X-ray structures or structural models of PM IV and cathepsin D, allowed an interpretation of the inhibition profiles of pepstatin A and its Tfm variants against these three enzymes. Interactions of the P1 side chain with amino acids that point into the S1 pocket appear to be critical for inhibitory activity. In summary, Tfm can be used to replace an isobutyl group and can affect the selectivity profile of a compound. These findings have implications for the design of novel bioactive molecules and synthetic mimics of natural compounds. PMID- 16307464 TI - Partially reversible adsorption of annexin A1 on POPC/POPS bilayers investigated by QCM measurements, SFM, and DMC simulations. AB - The kinetics of annexin A1 binding to solid-supported lipid bilayers consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)/1-palmitoyl-2-oleoyl sn-glycero-3-phosphoserine (POPS; 4:1) has been investigated as a function of the calcium ion concentration in the bulk phase. Quartz crystal microbalance measurements in conjunction with scanning force microscopy, fluorescence microscopy, and computer simulations indicate that at a given Ca2+ concentration annexin A1 adsorbs irreversibly on membrane domains enriched in POPS. By contrast, annexin A1 adsorbs reversibly on the POPC-enriched phase, which is composed of single POPS molecules embedded within a POPC matrix. The overall area occupied by the POPS-enriched phase is controlled by the CaCl2 concentration. Monte Carlo simulations suggest that the area of the POPS-enriched phase increases by a factor of 7 when the Ca2+ concentration is changed from 0.01 to 1 mM. PMID- 16307465 TI - Dynamics of formation of a helix-turn-helix structure in a membrane-active peptide: a time-resolved spectroscopic study. PMID- 16307466 TI - Regioselective carbon-carbon bond formation in proteins with palladium catalysis; new protein chemistry by organometallic chemistry. AB - Palladium-catalyzed reactions have contributed to the advancement of many areas of organic chemistry, in particular, the synthesis of organic compounds such as natural products and polymeric materials. In this study, we have used a Mizoroki Heck reaction for site-specific carbon-carbon bond formation in the Ras protein. This was performed by the following two steps: 1) the His6-fused Ras protein containing 4-iodo-L-phenylalanine at position 32 (iF32-Ras-His) was prepared by genetic engineering and 2) the aryl iodide group on the iF32-Ras-His was coupled with vinylated biotin in the presence of a palladium catalyst. The biotinylation was confirmed by Western blotting and liquid chromatography-mass spectrometry (LC MS). The regioselectivity of the Mizoroki-Heck reaction was furthermore confirmed by LC-MS/MS analysis. However, in addition to the biotinylated product (bF32-Ras His), a dehalogenated product (F32-Ras-His) was detected by LC-MS/MS. This dehalogenation resulted from the undesired termination of the Mizoroki-Heck reaction due to steric and electrostatic hindrance around residue 32. The biotinylated Ras showed binding activity for the Ras-binding domain as its downstream target, Raf-1, with no sign of decomposition. This study is the first report of an application of organometallic chemistry in protein chemistry. PMID- 16307467 TI - Mercury influences rat liver tyrosine aminotransferase activity and induction by dexamethasone. AB - The effects of mercury (Hg) on basal and dexamethasone-induced tyrosine aminotransferase (TAT) activity in rat liver were studied. Comparison of TAT activity after in vitro and in vivo mercury application revealed the influence of the metal only when applied in vivo, suggesting that the effects are expressed at the level of TAT gene transcription. Intraperitoneal administration of mercury at 1, 2 or 3 mg Hg kg(-1) b.w. 4 h before decapitation was shown to stimulate the basal activity of TAT. The most prominent increase was observed 4 h after the metal administration. When applied at 1 and 2 mg Hg kg(-1) b.w. mercury was also shown to reduce partially the extent of the enzyme induction by dexamethasone, which was injected intraperitoneally at 5 mg kg(-1) b.w. 5 h before death. The highest dose of mercury (3 mg Hg kg(-1) b.w.) almost completely abolished the dexamethasone effect. The finding that mercury increases basal activity of the enzyme while decreasing its induction by dexamethasone suggests that stimulatory effects of this metal on TAT activity are probably mediated by factors other than glucocorticoids. PMID- 16307468 TI - Effect of some acetylcholinesterase reactivators on human platelet aggregation in vitro. AB - Acetylcholinesterase (AChE) reactivators are employed for the prophylaxis and treatment of intoxications with organophosphorus AChE inhibitors, including nerve agents and pesticides. For the recovery of inhibited enzyme, derivatives from the group of pyridinium or bispyridinium aldoximes (called oximes) are used. Adverse effects of these substances are not well elucidated, because of their narrow and one-shot usage. Owing to this fact, the study evaluated the influence of some currently applied oximes on human platelet aggregation in vitro. The antiplatelet activity of pralidoxime, obidoxime, HI-6, methoxime and HLo 7 was assayed in human platelet rich plasma (2.5 x 10(8) platelets.ml(-1)) at a concentration of 1.35 mM. Arachidonic acid (AA), adenosine diphosphate (ADP), collagen (COL) and thrombin (TR) were used as agonists of platelet aggregation. All tested substances, except pralidoxime and methoxime, caused a significant inhibition of the aggregation process induced by AA, ADP and COL. Of the oximes assayed, none was found to influence TR triggered aggregation. Since reduced platelet aggregation can play an important role as an adverse effect in reactivator administration, further evaluation is needed for the estimation of the real impact of active oximes to the aggregation process in humans. PMID- 16307469 TI - A porcine model for fixed drug eruptions in humans: the case of antipyrine in the Yucatan micropig. AB - To date, there is no acceptable animal model to investigate fixed drug eruptions (FDEs) in humans. We briefly report here observations suggesting that the Yucatan micropig may be a useful animal model for that purpose. During an investigation of antipyrine absorption and disposition, we observed the development of FDEs after intravenous administration of a 1 g dose. Our observations were consistent with those reported in several investigations of humans taking a single dose of antipyrine. To confirm these results, a naive micropig was challenged. A male uncastrated Yucatan micropig (27.2 kg) was given a 1 g dose of antipyrine intravenously. After 30 days, this pig was rechallenged with the same intravenous dose of antipyrine (1 g). Blood samples were obtained to examine immunological endpoints. During the initial challenge, a fluid plaque (ca. 1-1.5 cm) appeared on the left hip of the pig ca. 6 h after dosing. After the rechallenge, inflamed pink patches were observed at the same sites where the blisters formed initially; however, no blisters re-formed. Changes of neutrophil, lymphocyte and eosinophil levels from baseline were noted 8 h after challenge. The micropig did not seem otherwise affected by the FDEs. These observations suggest that the Yucatan micropig, or swine in general, may be a useful animal model for detecting drugs that may cause FDEs in humans. PMID- 16307470 TI - Applicability of bioanalysis of multiple analytes in drug discovery and development: review of select case studies including assay development considerations. AB - The development of sound bioanalytical method(s) is of paramount importance during the process of drug discovery and development culminating in a marketing approval. Although the bioanalytical procedure(s) originally developed during the discovery stage may not necessarily be fit to support the drug development scenario, they may be suitably modified and validated, as deemed necessary. Several reviews have appeared over the years describing analytical approaches including various techniques, detection systems, automation tools that are available for an effective separation, enhanced selectivity and sensitivity for quantitation of many analytes. The intention of this review is to cover various key areas where analytical method development becomes necessary during different stages of drug discovery research and development process. The key areas covered in this article with relevant case studies include: (a) simultaneous assay for parent compound and metabolites that are purported to display pharmacological activity; (b) bioanalytical procedures for determination of multiple drugs in combating a disease; (c) analytical measurement of chirality aspects in the pharmacokinetics, metabolism and biotransformation investigations; (d) drug monitoring for therapeutic benefits and/or occupational hazard; (e) analysis of drugs from complex and/or less frequently used matrices; (f) analytical determination during in vitro experiments (metabolism and permeability related) and in situ intestinal perfusion experiments; (g) determination of a major metabolite as a surrogate for the parent molecule; (h) analytical approaches for universal determination of CYP450 probe substrates and metabolites; (i) analytical applicability to prodrug evaluations-simultaneous determination of prodrug, parent and metabolites; (j) quantitative determination of parent compound and/or phase II metabolite(s) via direct or indirect approaches; (k) applicability in analysis of multiple compounds in select disease areas and/or in clinically important drug-drug interaction studies. A tabular representation of select examples of analysis is provided covering areas of separation conditions, validation aspects and applicable conclusion. A limited discussion is provided on relevant aspects of the need for developing bioanalytical procedures for speedy drug discovery and development. Additionally, some key elements such as internal standard selection, likely issues of mass detection, matrix effect, chiral aspects etc. are provided for consideration during method development. PMID- 16307471 TI - Current approaches to unravel the mystery of dilated cardiomyopathy, a common cause of hereditary heart failure. PMID- 16307473 TI - Proinsulin lacking the A7-B7 disulfide bond, Ins2Akita, tends to aggregate due to the exposed hydrophobic surface. AB - A single mutation (C96Y) in the Ins2 gene, which disrupts the A7-B7 disulfide bond, causes the diabetic phenotype in Akita mice. We biochemically analyzed the conformation of wild-type and Akita mutant recombinant proinsulins. Gel filtration chromatography and dynamic light scattering revealed that the apparent size of the mutant proinsulin molecules was significantly larger than that of wild-type proinsulin, even in the absence of intermolecular disulfide bonds. Titration with a hydrophobic probe, 1-anilinonaphthalene-8-sulfonate, demonstrated that the mutant proinsulin was more hydrophobic than the wild type. In addition, circular dichroism studies revealed that the conformation of the mutant proinsulin was less stable than the wild type, which is consistent with the observation that hydrophobic residues are exposed on the surface of the proinsulin molecules. Studies with antiserum against the C-peptide of proinsulin indicated that the mutant proinsulin had an immunoreactivity that was at least one-tenth weaker than wild-type proinsulin, suggesting that the C-peptide of mutant proinsulin is buried inside the aggregate of the proinsulin molecule. These findings indicate that increased hydrophobicity of mutant proinsulin facilitates aggregate formation, providing a clue to the dominant negative effect in the Akita mouse. PMID- 16307474 TI - Macromolecular isoforms of Daphnia magna haemoglobin. AB - The haemoglobin (Hb) of Daphnia magna acclimated to different oxygen conditions was sampled, and in its natively assembled state it was separated by chromatofocusing. The Hb isoforms were analysed for their subunit composition under denaturating conditions by two-dimensional gel electrophoresis. The Hb system is suggested to consist of three predominant Hb aggregates, which are characterised by a specific subunit composition and synthesised in response to different ambient oxygen conditions. In normoxia, a dominant Hb aggregate (DmHbI) with a pI of 4.4-4.6 was composed of subunits B, C, E, F and G. In severe hypoxia, a different dominant Hb isoform (DmHbIII) with a pI of 5.7-5.9 was composed of subunits A, B, C, D, E and F. Further analyses in moderate hypoxia provided evidence for a third Hb isoform (DmHbII) composed of subunits B, C, D, E and F. Sequence alignment and homology modelling of the tertiary structure of the D. magna Hb domains 1 and 2 revealed functionally relevant substitutions of amino acid residues at positions B10, E7 and E11, which determine the functional properties of D. magna haemoglobin in terms of haem contact, oxygen binding and affinity. Both domains are predicted to possess the common haemoglobin fold, but helices C and D are not properly formed, and helix G is interrupted by a short coil. PMID- 16307475 TI - Rational engineering of a fluorescein-binding anticalin for improved ligand affinity. AB - The anticalin FluA is an artificial lipocalin with novelspecificity for the fluorescein group, which was engineered from an insect bilin-binding protein by targeted random mutagenesis and selection. Based on the crystal structure of FluA, an attempt was made to improve the complementarity of its ligand pocket to fluorescein by rational protein design. Several side chains participating in sub optimal interactions with the ligand were identified and replaced by residues that promised a better steric fit. As a result, the substitution of Ala45 by Ile and of Ser114 by Thr or Arg led to a tight affinity of ca. 1 nM, which is approximately 30-fold better than that of the parental anticalin. Similar to the original FluA, the improved version shows almost complete quenching of the bound ligand fluorescence. Interestingly, the quenching effect was significantly reduced when Trp129 was replaced by Tyr, thus supporting the previously postulated role of this residue, which closely packs against the bound ligand, for efficient electron transfer to the excited fluorescein. Circular dichroism spectra revealed that all variants investigated had retained the lipocalin fold. Corresponding thermal unfolding experiments confirmed similar folding stabilities, with melting temperatures ranging from 52.9 to 60.5 degrees C (i.e., for the high-affinity variant). PMID- 16307476 TI - Monitoring the real-time kinetics of the hydrolysis reaction of guanine nucleotide-binding proteins. AB - The conversion of guanosine triphosphate (GTP) to guanosine diphosphate (GDP) and inorganic phosphate (Pi) by guanine nucleotide-binding proteins (GNBPs) is a fundamental enzyme reaction in living cells that acts as an important timer in a variety of biological processes. This reaction is intrinsically slow but can be stimulated by GTPase-activating proteins (GAPs) by several orders of magnitude. In the present study, we synthesized and characterized a new fluorescent nucleotide, 2'(3')-O-(N-ethylcarbamoyl-(5''-carboxytetramethylrhodamine) amide) GTP, or tamraGTP, which is sensitive towards conformational changes of certain GNBPs induced by GTP hydrolysis. Unlike other fluorescent nucleotides, tamra-GTP allows real-time monitoring of the kinetics of the intrinsic and GAP-catalyzed GTP hydrolysis reactions of small GNBPs from the Rho family. PMID- 16307477 TI - ClpV, a unique Hsp100/Clp member of pathogenic proteobacteria. AB - Hsp100/Clp proteins are key players in the protein quality control network of prokaryotic cells and function in the degradation and refolding of misfolded or aggregated proteins. Here we report the identification of a new class of Hsp100/Clp proteins, termed ClpV (virulent strain), that are present in bacteria interacting with eukaryotic cells, including human pathogens. The ClpV proteins are most similar to ClpB proteins within the Hsp100/Clp family, but cluster in a separate phylogenetic tree with a remarkable distance to ClpB. ClpV representatives from Salmonella typhimurium and enteropathogenic Escherichia coli form oligomeric assemblies and display ATP hydrolysis rates comparable to ClpB. However, unlike ClpB, both ClpV proteins failed to solubilize aggregated proteins. This lack of disaggregation activity correlated with the inability of ClpB model substrates to stimulate the ATPase activity of ClpV proteins, indicating differences in substrate selection. Furthermore, we show that clpV genes are generally organized in a conserved gene cluster, encoding a potential secretion system, and we demonstrate that increased levels of a dominant negative variant of either S. typhimurium or Yersinia pseudotuberculosis ClpV strongly reduce the ability of these pathogenic bacteria to invade epithelial cells. We propose a role of this novel and unique class of AAA+ proteins in bacteria-host cell interactions. PMID- 16307478 TI - Plasmodium falciparum 2-Cys peroxiredoxin reacts with plasmoredoxin and peroxynitrite. AB - Thioredoxin peroxidase 1 (TPx1) of the malarial parasite Plasmodium falciparum is a 2-Cys peroxiredoxin involved in the detoxification of reactive oxygen species and - as shown here - of reactive nitrogen species. As novel electron acceptor of reduced TPx1, we characterised peroxynitrite; the rate constant for ONOO- reduction by the enzyme (1 x 10(6) M(-1) s(-1) at pH 7.4 and 37 degrees C) was determined by stopped-flow measurements. As reducing substrate of TPx1, we identified - aside from thioredoxin - plasmoredoxin; this 22-kDa protein occurs only in malarial parasites. When studying the potential roles of Cys74 and Cys170 of Tpx1 in catalysis, as well as in oligomerisation behaviour, we found that replacement of Cys74 by Ala influenced neither the dimerisation nor enzymatic activity of TPx1. In the C170A mutant, however, the kcat/Km for reduced Trx as a substrate was shown to be approximately 50-fold lower and, in contrast to the wild-type enzyme, covalently linked dimers were not formed. For the catalytic cycle of TPx1, we conclude that oxidation of the peroxidatic Cys50 by the oxidising substrate is followed by the formation of an intermolecular disulfide bond between Cys50 and Cys170' of the second subunit, which is then attacked by an external electron donor such as thioredoxin or plasmoredoxin. PMID- 16307479 TI - Lysophosphatidic acid affinity chromatography reveals pyruvate kinase as a specific LPA-binding protein. AB - Lysophosphatidic acid is a pleiotropic lipid signaling molecule that evokes a broad array of cellular responses including proliferation, tumor cell invasion, neurite retraction, cytoskeletal rearrangements and smooth muscle contraction. Generally, lysophosphatidic acid triggers physiological responses through interaction with specific plasma membrane receptors called LPA 1-4. There is, however, increasing evidence in support of intracellular proteins that interact with LPA. We employed Affigel-immobilized LPA to isolate cytoplasmic proteins that interact with this lysophospholipid. Among the proteins retained by this affinity matrix, pyruvate kinase, clathrin heavy chain and heat shock protein 70 (Hsp70) were identified by mass spectrometry. Isothermal titration calorimetry showed that pyruvate kinase contains one binding site for LPA (Ka approx. 10(6) M(-1)). Furthermore, LPA dissociates enzymatically active pyruvate-kinase tetramers into less active dimers, and is maximally active at concentrations close to its critical micelle concentration. These effects were not mimicked by other lysophospholipids. Co-immunoprecipitation experiments showed that pyruvate kinase interacts with clathrin, and confocal imaging revealed co-localization between clathrin and pyruvate kinase in the perinuclear region of cells. Our data suggest that pyruvate kinase partly exists in complex with clathrin in subcellular membranous areas, and that locally increased LPA levels can trigger inactivation of the metabolic enzyme. PMID- 16307480 TI - The polysaccharide scaffold of PrP 27-30 is a common compound of natural prions and consists of alpha-linked polyglucose. AB - An inert polysaccharide scaffold identified as a 5-15% component of prion rods (PrP 27-30) is unambiguously distinguishable from the N-glycosyl groups and the GPI anchor of PrP, and consists predominantly of 1,4-linked glucose with some branching via 1,4,6-linked glucose. We show that this polysaccharide scaffold is a common secondary component of prions found in hamster full-length PrP(Sc), prion rods and in mouse ScN2a prions from cell culture. The preparation from prion rods was improved, resulting in a polysaccharide scaffold free of remaining infectivity. Furthermore, we determined the stereochemistry of the glycoside linkages as pre-dominantly if not entirely alpha-glycosidic. The origin of the polysaccharide, its interaction with PrP and its potential relation to glycogen and corpora amylacea are discussed. PMID- 16307481 TI - Cytosolic persistence of mouse brain CYP1A1 in chronic heme deficiency. AB - Previous work has demonstrated that the function of extrahepatic cytochrome P450 CYP1A1 is dependent on the availability of heme. CYP1A1 is involved in the activation of polyaromatic hydrocarbons. In the present study we used a transgenic mouse model with chronic impairment of heme synthesis - female porphobilinogen deaminase-deficient (PBGD-/-) mice - to investigate the effects of limited heme in untreated and beta-naphthoflavone (beta-NF)-treated animals on the function of CYP1A1 in brain. The heme content of PBGD-/- mice was diminished in the liver and brain compared to wild types. In the liver, partial heme deficiency led to less potent induction of CYP1A1 mRNA after beta-NF treatment. In the brain, CYP1A1 protein was detected not only at the endoplasmic reticulum (ER), but also in the cytosol of PBGD-/- mice. Furthermore, 7-deethylation of ethoxyresorufin, an indicator of CYP1A1 metabolic activity, could be restored by heme in cytosol of PBGD-/- mouse brain. Independent of the genotype, we found only one cyp1a1 gene product, indicating that the cytosolic appearance of CYP1A1 most likely did not originate from mutant alleles. We conclude that heme deficiency in the brain leads to incomplete heme saturation of CYP1A1, which causes its improper incorporation into the ER membrane and persistence in the cytosol. It is suggested that diseases caused by relative heme deficiency, such as hepatic porphyrias, may lead to impaired hemoprotein function in brain. PMID- 16307482 TI - B-Raf and C-Raf signaling investigated in a simplified model of the mitogenic kinase cascade. AB - Signaling pathways based on the reversible phosphorylation of proteins control most aspects of cellular life in higher organisms. Extracellular stimuli can induce growth, differentiation, survival and the stress response through a number of highly conserved signaling pathways. We discuss how the intensity and duration of signals may have dramatic consequences on the way cells respond to stimuli. Picking the central Ras-Raf-MEK-ERK signal cascade, we developed a mathematical model of how stimuli induce different signal patterns and thereby different cellular responses, depending on cell type and the ratio between B-Raf and C-Raf. Based on biochemical data for activation and dephosphorylation, as well as the differential equations of our model, we suggest a different signaling pattern and response result for B-Raf (strong activation, sustained signal) and C-Raf (steep activation, transient signal). We further support the significance of such differential modulatory signaling by showing different Raf isoform expression in various cell lines and experimental testing of the predicted kinase activities in B-Raf, C-Raf and mutated versions. PMID- 16307483 TI - Inhibition of human kallikreins 5 and 7 by the serine protease inhibitor lympho epithelial Kazal-type inhibitor (LEKTI). AB - LEKTI is a 120-kDa protein that plays an important role in skin development, as mutations affecting LEKTI synthesis underlie Netherton syndrome, an inherited skin disorder producing severe scaling. Its primary sequence indicates that the protein consists of 15 domains, all resembling a Kazal-type serine protease inhibitor. LEKTI and two serine proteases belonging to the human tissue kallikrein (hK) family (hK5 and hK7) are expressed in the granular layer of skin. In this study, we characterize the interaction of two recombinant LEKTI fragments containing three or four intact Kazal domains (domains 6-8 and 9-12) with recombinant rhK5, a trypsin-like protease, and recombinant rhK7, a chymotrypsin like protease. Both fragments inhibited rhK5 similarly in binding and kinetic studies performed at pH 8.0, as well as pH 5.0, the pH of the stratum corneum where both LEKTI and proteases may function. Inhibition equilibrium constants (Ki) measured either directly in concentration-dependent studies or calculated from measured association (kass) and dissociation (kdis) rate constants were 1.2 5.5 nM at pH 8.0 and 10-20 nM at pH 5.0. At pH 8.0, kass and kdis values were 4.7 x 10(5) M(-1) s(-1) and 5.5 x 10(-4) s(-1), and at pH 5.0 they were 4.0 x 10(4) M(-1) s(-1) and 4.3 x 10(-4) s(-1), respectively. The low Ki and kdis values (t1/2 of 20-25 min) indicate tight and specific association. Only fragment 6-9' was a good inhibitor of rhK7, demonstrating a Ki of 11 nM at pH 8.0 in a reaction that was rapidly reversible. These results show that LEKTI, at least in fragment form, is a potent inhibitor of rhK5 and that this protease may be a target of LEKTI in human skin. PMID- 16307484 TI - Cloning, expression and characterization of Bauhinia variegata trypsin inhibitor BvTI. AB - A Bauhinia variegata trypsin inhibitor (BvTI) cDNA fragment was cloned into the pCANTAB5E phagemid. The clone pAS 1.1.3 presented a cDNA fragment of 733 bp, including the coding region for a mature BvTI protein comprising 175 amino acid residues. The deduced amino acid sequence for BvTI confirmed it as a member of the Kunitz-type plant serine proteinase inhibitor family. The BvTI cDNA fragment encoding the mature form was cloned into the expression vector, pET-14b, and ex pressed in E. coli BL21 (DE3) pLysS in an active form. In addition, a BvTI mutant form, r(mut)BvTI, with a Pro residue as the fifth amino acid in place of Leu, was produced. The recombinant proteins, rBvTI and r(mut)BvTI, were purified on a trypsin-Sepharose column, yielding 29 and 1.44 mg/l of active protein, respectively, and showed protein bands of approximately 21.5 kDa by SDS-PAGE. Trypsin inhibition activity was comparable for rBvTI (Ki=4 nM) and r(mut)BvTI (Ki=6 nM). Our data suggest that the Leu to Pro substitution at the fifth amino terminal residue was not crucial for proteinase inhibition. PMID- 16307485 TI - Recombinant human cathepsin X is a carboxymonopeptidase only: a comparison with cathepsins B and L. AB - The S1 and S2 subsite specificity of recombinant human cathepsins X was studied using fluorescence resonance energy transfer (FRET) peptides with the general sequences Abz-Phe-Xaa-Lys(Dnp)-OH and Abz-Xaa-Arg-Lys(Dnp)-OH, respectively (Abz=ortho-aminobenzoic acid and Dnp=2,4-dinitrophenyl; Xaa=various amino acids). Cathepsin X cleaved all substrates exclusively as a carboxymonopeptidase and exhibited broad specificity. For comparison, these peptides were also assayed with cathepsins B and L. Cathepsin L hydrolyzed the majority of them with similar or higher catalytic efficiency than cathepsin X, acting as an endopeptidase mimicking a carboxymonopeptidase (pseudo-carboxymonopeptidase). In contrast, cathepsin B exhibited poor catalytic efficiency with these substrates, acting as a carboxydipeptidase or an endopeptidase. The S1' subsite of cathepsin X was mapped with the peptide series Abz-Phe-Arg-Xaa-OH and the enzyme preferentially hydrolyzed substrates with hydrophobic residues in the P1' position. PMID- 16307486 TI - Purification of Mg2+-dependent phosphatidate phosphohydrolase from rat liver: new steps and aspects. AB - A new procedure for the partial purification of Mg2+-dependent, N-ethylmaleimide sensitive phosphatidate phosphohydrolase (Mg2+-PAP; EC 3.1.3.4) from rat liver cytosol is described, using protein precipitation with MgCl2, gel filtration on Sephacryl S-400, chromatography on DEAE-cellulose and affinity chromatography on calmodulin-agarose. From the parallel change in staining intensity and in the level of the specific activity of enzyme fractions, a relationship between a 90 kDa SDS gel band, identified as the beta-isoform of the 90-kDa heat shock protein, and Mg2+-PAP could be detected. PMID- 16307487 TI - 17-AAG: mechanisms of antitumour activity. AB - Heat-shock protein 90 (Hsp90) is a molecular chaperone involved in three dimensional folding, intracellular translocation and degradation of multiple key regulatory proteins. Accumulated evidence has indicated an important role of Hsp90 in several signal transduction pathways that are deregulated in carcinogenesis. 17-allylamino-17-demethoxygeldanamycin (17-AAG), a selective inhibitor of Hsp90, is currently under clinical investigation in advanced malignancies in which Hsp90 client proteins are implicated. This article discusses the mechanistic evidence underlying 17-AAG's cytostatic, proapoptotic, antiangiogenic and anti-invasive properties that provide the basis for its antitumour activity and underscores its unique therapeutic potential as a multi targeted agent, as opposed to most of the current-generation molecular therapeutics. PMID- 16307488 TI - Targeting integrins for the control of tumour angiogenesis. AB - The crucial role of cell extracellular matrix communication in angiogenesis is well established; thus, it is not surprising that integrins have gained considerable attention as targets for the treatment of neovascular disease. Given the diversity of ligands and complexity of integrin signalling, a new appreciation for the divergent roles of integrins in angiogenesis is emerging. It is becoming clear that integrins regulate angiogenesis in both a positive and negative manner. New studies have provided a better understanding of integrin structure as it relates to ligand binding and signalling. This new insight has opened exciting possibilities for the design of novel inhibitors for clinical applications. In this review, studies concerning the cooperative interactions between integrins and regulatory molecules and possible new strategies for controlling angiogenesis will be discussed. PMID- 16307489 TI - The glutamatergic system outside the CNS and in cancer biology. AB - Glutamate is a major excitatory neurotransmitter in the CNS. The signalling machinery consists of: glutamate receptors, which are responsible for signal input; plasma glutamate transporters, which are responsible for signal termination; and vesicular glutamate transporters for signal output through exocytic release. Recently, data have suggested that the glutamatergic system plays an important role in non-neuronal tissues. In addition, the expression of glutamatergic system has been implicated in tumour biology. This review outlines the evidence, which suggests that the glutamatergic system may have an important role in cancer biology. PMID- 16307490 TI - Histone deacetylase inhibitors: discovery and development as anticancer agents. AB - Histone deacetylase (HDAC) inhibitors are a new class of targeted anticancer agents. Several HDAC inhibitors are in clinical trials and have shown significant activity against a spectrum of both haematological and solid tumours at doses that are well tolerated by patients. HDACs and histone acetyltransferases can, by reversible acetylation, modify the structure and function of histones and proteins in transcription factor complexes, which are involved in the regulation of gene expression, as well as many non-histone proteins that are involved in regulating cell proliferation and cell death. HDAC inhibitors are a structurally diverse group of molecules; these agents selectively alter the expression of genes. HDAC inhibitors can induce cancer cell death, whereas normal cells are relatively resistant to HDAC inhibitor-induced cell death. PMID- 16307491 TI - Triclabendazole for the treatment of fascioliasis and paragonimiasis. AB - Fascioliasis and paragonimiasis, which are caused by liver flukes (Fasciola) and lung flukes (Paragonimus), are emerging public health problems. Several hundred millions of people are at risk of the two diseases that cause considerable morbidity and delay socio-economic development. Triclabendazole, a benzimidazole derivative, has been routinely used since 1983 in veterinary medicine to control infections with Fasciola spp. in domestic herbivorous animals. In 1986, a veterinary formulation of triclabendazole was first administered to two human patients with fascioliasis. Clinical data obtained thus far suggest that triclabendazole is the most efficacious and best tolerated drug for the treatment of fascioliasis. Moreover, the drug holds promise as a useful therapeutic alternative to praziquantel in the treatment of paragonimiasis. This review of triclabendazole includes an overview of the pharmacodynamics and pharmacokinetics, toxicology and efficacy against the major food-borne trematodes in laboratory animals. Data from case reports and clinical trials in humans infected with Fasciola spp. and Paragonimus spp. are summarised and the current state of triclabendazole regarding treatment of human fascioliasis and paragonimiasis is described. Efforts to facilitate broader registration of this drug should go hand-in-hand with research and development on novel drugs against food-borne trematodiasis, better access to improved sanitation, sound health education and the development of new technologies for assuring food safety. PMID- 16307492 TI - New and improved glucocorticoid receptor ligands. AB - The therapeutic and prophylactic use of glucocorticoids is widespread due to their powerful anti-inflammatory, antiproliferative and immunomodulatory activity. However, long-term use of these drugs can result in severe dose limiting side effects. One of the most critical and debilitating side effects is osteoporosis, which leads to increased risk of fractures. Glucocorticoids damage bone through several different mechanisms. The search for novel glucocorticoids that have reduced side effects in bone and other tissues is being driven by the identification of new mechanisms of action of the glucocorticoid receptor. This may facilitate the detection of new, safer therapies with efficacies equivalent to currently prescribed steroids. PMID- 16307493 TI - The protein kinase C-beta inhibitor, ruboxistaurin, for the treatment of diabetic microvascular complications. AB - The potential for addressing the underlying biological abnormalities in diabetes has eluded most investigators because of the complex mechanisms underlying the effects of diabetes on the pathogenesis of the complications. Although macrovascular complications, such as myocardial infarction, stroke and gangrene, are only partially attributable to hyperglycaemia and its attendant effects, the microvascular complications including retinopathy, nephropathy and neuropathy are directly related to the degree of hyperglycaemia. In controlled trials, a 22-34% reduction in one of these side effects was achieved for every 1% reduction in glycosylated haemoglobin. Theoretically, it should be feasible to eliminate these complications in a perfect world. However, achieving euglycaemia is nearly impossible and there is increasing data to suggest that it may be an elusive target with ever lower levels being implicated in the pathogenesis of microvascular disease and there is a price to be paid of hypoglycaemia if further intensification is pursued. A logical argument would be to block pathways that are activated by hyperglycaemia. A major pathway implicated is the activation of protein kinase C-beta in all of the targeted tissues, and there is animal data to support the notion that blocking this pathway can reverse or abrogate the untoward effects of diabetes. The possible role of the protein kinase C-beta inhibitor, ruboxistaurin, in animal studies and the recently reported clinical studies to place in perspective a possible addition to the therapeutic armamentarium of the imperfect world of diabetes control will be reviewed. PMID- 16307494 TI - Potential of growth hormone-releasing hormone antagonists (JV-1-36 and JV-1-42) for the treatment of brain tumours. PMID- 16307495 TI - Government action needed to step up research and development for world's most neglected diseases. PMID- 16307498 TI - Atovaquone/proguanil for the prophylaxis and treatment of malaria. AB - Increases in international travel and escalating drug resistance have resulted in a growing number of travelers at risk of contracting malaria. Drug resistance and intolerance to standard agents such as chloroquine, sulfadoxine/pyrimethamine and mefloquine has highlighted the need for new antimalarials. The recently licensed fixed combination of atovaquone and proguanil hydrochloride (Malarone) is a promising new agent to prevent and treat Plasmodium falciparum malaria. Randomized controlled trials have shown that atovaquone/proguanil is well tolerated and efficacious for the prevention and treatment of drug-resistant P. falciparum malaria. Atovaquone/proguanil is active against the liver stage of P. falciparum malaria parasites and when used as a prophylactic agent it can be discontinued shortly after leaving malaria-endemic areas, offering a clear advantage for drug adherence. PMID- 16307499 TI - Rabeprazole: the role of proton pump inhibitors in Helicobacter pylori eradication. AB - Proton pump inhibitors have become one of the cornerstones in the treatment of Helicobacter pylori infection. Rabeprazole (Pariet) is a substituted benzimidazole proton pump inhibitor with potent gastric acid suppression properties. Its high acid-base dissociation constant allows activation over a broader pH range, resulting in quick, irreversible binding to the H+/K+-ATPase pump, and a more rapid onset of action compared with omeprazole, lansoprazole and pantoprazole. Unlike other proton pump inhibitors, the metabolism of rabeprazole is primarily via a nonenzymatic reduction to the thioether derivative, and the cytochrome P450 isoenzyme 2C19 is only partly involved in its metabolism. The effect of genetic polymorphism in cytochrome P450 isoenzyme 2C19 on the pharmacokinetics and pharmacodynamics of rabeprazole is therefore limited. In humans, once-daily dosing of 5-40 mg of rabeprazole inhibits gastric acid secretion in a dose-dependent manner. In vitro studies have shown that rabeprazole possesses more potent antibacterial properties against the growth of H. pylori than other proton pump inhibitors. Furthermore, its thioether derivative has more potent inhibitory in vitro activity against the growth and motility of clarithromycin-resistant H. pylori than other proton pump inhibitors or commonly used antimicrobials. Despite these inherent favorable characteristics of rabeprazole, randomized controlled trials have largely shown equivalence amongst proton pump inhibitors when used with two antibiotics in the eradication of H. pylori, with cure rates of 75-89% on an intent-to-treat basis. However, rabeprazole appears to consistently achieve such comparable eradication rates even when used at reduced doses (10 mg twice daily) as part of clarithromycin based triple therapy. PMID- 16307500 TI - Fixed dose combination abacavir/lamivudine in the treatment of HIV-1 infection. AB - The fixed dose combination of abacavir with lamivudine represents a new treatment option for patients infected with HIV. Fixed dose combination abacavir/lamivudine has the convenience of one pill and once-daily dosing. It achieves comparable suppression of plasma HIV RNA with the pill's individual components dosed twice daily and with thymidine analogs combined with lamivudine. The combination is well tolerated, with the potential advantages of less lipoatrophy and fewer metabolic perturbations. However, the abacavir component may cause hypersensitivity reactions, which are reported in up to 8% of patients, and are potentially life threatening. Fixed dose combination abacavir/lamivudine should be considered as a viable treatment option for HIV-infected patients, particularly for those who have otherwise limited nucleoside reverse transcriptase inhibitor choices. PMID- 16307501 TI - Thymalfasin for the treatment of chronic hepatitis C infection. AB - Approximately 50% of treatment-naive hepatitis C patients fail to achieve a sustained virologic response with standard peginterferon and ribavirin therapy. Patients who are infected with genotype 1 have high viral loads and are nonresponders to previous therapy, and are even more difficult to treat, underscoring the need for new therapeutic options. Thymalfasin (thymosin-alpha1), in combination with peginterferon-alpha2a, has demonstrated efficacy among difficult-to-treat patients with hepatitis C. The addition of ribavirin to thymalfasin and peginterferon-alpha2a has also exhibited promising results among patients who have genotype 1 hepatitis C, high viral loads and are nonresponders to previous therapy. PMID- 16307502 TI - Management of healthcare-associated methicillin-resistant Staphylococcus aureus. AB - Healthcare-associated methicillin-resistant Staphylococcus aureus is a major cause of nosocomial infections worldwide, with significant attributable morbidity and mortality in addition to pronounced healthcare costs. Treatment results with vancomycin--the current recommended antibiotic for serious methicillin-resistant S. aureus infections--have not been impressive. The recent availability of effective antimicrobial agents other than glycopeptides, such as linezolid and daptomycin, as well as the anticipated approval of newer agents with diverse mechanisms of action, has somewhat ameliorated the threat posed by this organism. However, these drugs are expensive, and there is still no overall satisfactory strategy for reducing the incidence of healthcare-associated methicillin resistant S. aureus in endemic regions. Although early results with the Society for Healthcare Epidemiology of America guidelines give cause for cautious optimism, long-term experience is lacking, and it is likely that these guidelines will have to be adapted according to local conditions and resources before implementation. Trends to keep in mind when considering the problem of healthcare associated methicillin-resistant S. aureus include the advent of community associated methicillin-resistant S. aureus, and the propensity of S. aureus to evolve and acquire resistance determinants over time. This was last vividly demonstrated by the handful of vancomycin-resistant S. aureus isolated recently, which had acquired the vancomycin resistance gene from vancomycin-resistant enterococci. PMID- 16307503 TI - The search for new antimicrobials: why we need new options. AB - The increasing identification of antibiotic-resistant pathogens that cause serious infections cannot be ignored. Although the future cannot be predicted with certainty, it is surely possible that an extensive epidemic of resistant bacterial infections could potentially harm millions of people. Given that it takes more than 10 years to establish the efficacy and safety of new compounds, there is an urgent need to restock the antibiotic pipeline. Only a few new antibacterial agents have received approval by the US Food and Drug Administration in the last 5 years, including linezolid in 2001, cefditoren, pivoxil and ertapenem in 2002, gemifloxacin and daptomycin in 2003, and telithromycin in 2004. Many of these agents are improved derivatives from established classes of antibiotics, and several are directed primarily at resistant Gram-positive bacteria (e.g., linezolid and daptomycin). One promising new addition is the recent approval of tigecycline (Tigacyl, Wyeth) in June 2005. PMID- 16307504 TI - Mechanisms of resistance to beta-lactams in some common Gram-negative bacteria causing nosocomial infections. AB - Gram-negative bacilli remain major killers of hospitalized patients and continue to evolve new resistance mechanisms. This review describes the mechanisms of resistance to beta-lactam antibiotics from those Gram-negative pathogens most often isolated from nosocomial infections. PMID- 16307505 TI - Macrolide antibiotics and bronchiolitis obliterans following lung transplantation. AB - The anti-inflammatory effects of macrolide antibiotics are of emerging importance. Over the past 20 years, chronic inflammatory respiratory conditions, including diffuse panbronchiolitis, cystic fibrosis and asthma have benefited from long-term low-dose macrolide therapy. Obliterative bronchiolitis, a form of chronic allograft dysfunction in lung transplant recipients, has been reported to be a condition in which macrolide therapy may be indicated. A number of recent cohort studies have been encouraging. Disease progression was delayed with macrolide antibiotic therapy as patients showed a sustained improvement in pulmonary function tests. Up to now, obliterative bronchiolitis has been resistant to most forms of proposed treatment. This review aims to highlight the etiology of obliterative bronchiolitis, the mechanisms of immunomodulation associated with macrolide therapy and how macrolide therapy may be effective by way of these possible mechanisms. PMID- 16307506 TI - Development of effective therapies against West Nile virus infection. AB - Since its entry into North America in 1999, West Nile virus has spread throughout the USA and Canada, and now annually causes a clinical spectrum of human disease ranging from a self-limiting acute febrile illness to potentially lethal encephalitis. Although no therapy is currently approved for use in humans, several strategies are being pursued to develop effective prophylaxis and treatments. This review describes the epidemiology, clinical presentation and pathogenesis of West Nile virus infection, and highlights recent progress towards an effective therapy. PMID- 16307507 TI - Prospects of antiviral and anti-inflammatory therapy for respiratory syncytial virus infection. AB - Respiratory syncytial virus is the leading viral cause of death in children less than 2 years of age, and is an increasing cause of morbidity and mortality in transplant patients and the elderly. Respiratory syncytial virus causes upper and lower respiratory tract infections, which can lead to severe bronchiolitis and pneumonia. High-risk groups for severe respiratory syncytial virus infection include infants with a history of premature birth with or without chronic lung disease, children with congenital heart disease, children with cystic fibrosis or chronic lung diseases, and immunosuppressed patients or patients with immunodeficiency. However, the majority of infants who have severe respiratory syncytial virus disease are born at full term and are otherwise healthy. It is unclear why children, the elderly and the immunosuppressed are at much higher risk for severe disease; however, a respiratory syncytial virus-induced immune pathologic mechanism has long been suspected. Attempts to develop a safe and effective vaccine against respiratory syncytial virus have failed. Antirespiratory syncytial virus immunotherapy, although effective prophylactically, does not provide any beneficial clinical outcome when administered therapeutically, indicating that respiratory syncytial virus-induced pathology is most likely the result of the inflammatory response to infection, rather than a direct viral cytopathic effect. Thus, a combined antiviral and anti inflammatory therapy may represent the safest and most efficient treatment for acute respiratory syncytial virus infection. In this review, the current knowledge that has set the rationale for the development of such therapy is summarized. PMID- 16307508 TI - Use of rapid HIV antibody testing for controlling the HIV pandemic. AB - The HIV pandemic continues to expand throughout Africa and southern Asia. Despite recent advances in therapy, the primary means of prevention continues to be the identification of infected patients through diagnostic testing, and the provision of counseling services to reduce HIV transmission. In order to facilitate the identification of infected patients, great strides have been made during the past 10 years towards the development of simple, rapid HIV antibody assays that require no specialized equipment, are relatively stable at ambient temperatures and can be easily performed by people who do not have a laboratory background. Evaluations of these assays have shown that when used in multiple assay algorithm strategies, they perform comparably with current laboratory-based methods. Effective global implementation of these tests will require a concerted effort from public and private health agencies, however, expanding the use of these assays can provide a significant opportunity to slow the devastating spread of HIV. PMID- 16307509 TI - Prevention of mother-to-child transmission of HIV: treatment options. AB - Mother-to-child transmission of HIV-1 is responsible for 1800 new infections in children daily. The use of antiretroviral therapy can significantly reduce the risk of transmission. In settings where highly active antiretroviral therapy is available, mother-to-child transmission rates have been reduced to less than 2%, in the absence of breastfeeding. Women who require ongoing highly active antiretroviral therapy for their own health should receive this in pregnancy, which is also very effective in preventing transmission. Where resources allow, combination highly active antiretroviral therapy can also be used for preventing mother-to-child transmission in those women who do not yet need to receive ongoing treatment. The potential side effects of highly active antiretroviral therapy must be considered in pregnant women and their infants. Where highly active antiretroviral therapy is not possible, a dual combination regimen of antepartum zidovudine with single-dose nevirapine to mother and baby can reduce transmission to below 5%. In many places, the only available option is single dose nevirapine to mother and baby, which is effective in halving transmission risk, although the effectiveness in practice will be influenced by continued infection through breastfeeding, and by program factors such as the uptake of HIV testing. Exposure to nevirapine for mother-to-child transmission prevention can select for resistant virus in the majority of women. While the long-term implications of this are not completely clear, this selection can be reduced by the addition of short courses of postpartum zidovudine and lamivudine. PMID- 16307510 TI - Diagnosis of latent Mycobacterium tuberculosis infection: is the demise of the Mantoux test imminent? AB - Tuberculosis is responsible for more then 2 million deaths worldwide each year and vies with HIV as the world's most fatal infectious disease. In many developing countries, attempts to control the spread of infection rely solely on identification and treatment of those with active disease, ignoring subclinical infection. However, in developed countries, large efforts are also expended to identify and give prophylactic drugs to people with latent tuberculosis infection. Until recently, the 100-year-old tuberculin skin test (Mantoux) has been the only available diagnostic test for latent tuberculosis infection, despite its many well-known limitations. Advances in scientific knowledge have led to the development of tests for tuberculosis that measure the production of interferon-gamma by T-cells stimulated in vitro with Mycobacterium tuberculosis specific antigens. These interferon-gamma tests are highly specific and unaffected by prior Bacille Calmette-Guerin vaccination or immune reactivity to most atypical mycobacteria. They are more sensitive than the tuberculin skin test in detecting people with active tuberculosis, and their results correlate more closely with M. tuberculosis exposure risk factors than the tuberculin skin test in people likely to have latent tuberculosis infection. Science has caught up with one of the oldest diagnostic tests still in use worldwide, and the adoption of new, tuberculosis-specific interferon-gamma-based tests should move us one step closer to better control of this insidious pathogen. PMID- 16307511 TI - Drug-resistant tuberculosis: controversies and challenges in pediatrics. AB - Tuberculosis remains one of the top two causes of death caused by a single infectious disease worldwide, despite curative therapy. Children with tuberculosis are especially difficult to detect, since acid fast bacilli smears and cultures are usually negative and clinical signs are nonspecific or lacking. Multidrug-resistant tuberculosis, or tuberculosis resistant to at least isoniazid and rifampin, has emerged in most areas of the world over the past 20 years. Treatment of multidrug-resistant tuberculosis is more expensive and difficult. The second-line tuberculosis medications required for treatment are more toxic and less efficacious than standard treatment. These medications are not readily available in many areas of the world where drug resistance is most common. Fluoroquinolones are one of the most promising classes of second-line medications, but are not generally recommended for use in children. Ethambutol is recommended in the initial treatment of tuberculosis in children treated in areas where there is a risk of drug-resistant disease and the susceptibility of the source case is not known. Some experts have been hesitant to use ethambutol due to the risk of visual impairment associated with the drug and the difficulties in monitoring vision in young children. Pediatric drug formulations are not available for most antituberculosis medications, even the first-line tuberculosis drugs. Treatment of children exposed, infected or ill with multidrug-resistant tuberculosis is reviewed with special emphasis on second-line drugs, including recommended dosage, available formulations and necessary monitoring. While new cases of multidrug-resistant tuberculosis have decreased in most developed countries over the past 10 years, cases continue to increase in many developing countries and among immigrants from high-risk areas. Tuberculosis and multidrug resistant tuberculosis are serious threats requiring worldwide strategies to control and treat. Better diagnostic tests, medications, public health strategies and vaccines will all be needed to eliminate tuberculosis. PMID- 16307513 TI - First meeting on Developments in Protein Interaction Analysis (DiPIA 2005). August 28-31, 2005, PA, USA. PMID- 16307514 TI - 2005 Biomarker Discovery Summit: impressions and summary. September 26-28, 2005, PA, USA. PMID- 16307515 TI - Bioinformatics approaches in clinical proteomics. AB - Protein expression profiling is increasingly being used to discover, validate and characterize biomarkers that can potentially be used for diagnostic purposes and to aid in pharmaceutical development. Correct analysis of data obtained from these experiments requires an understanding of the underlying analytic procedures used to obtain the data, statistical principles underlying high-dimensional data and clinical statistical tools used to determine the utility of the interpreted data. This review summarizes each of these steps, with the goal of providing the nonstatistician proteomics researcher with a working understanding of the various approaches that may be used by statisticians. Emphasis is placed on the process of mining high-dimensional data to identify a specific set of biomarkers that may be used in a diagnostic or other assay setting. PMID- 16307516 TI - Classification and identification of bacteria using mass spectrometry-based proteomics. AB - Timely classification and identification of bacteria is of vital importance in many areas of public health. Mass spectrometry-based methods provide an attractive alternative to well-established microbiologic procedures. Mass spectrometry methods can be characterized by the relatively high speed of acquiring taxonomically relevant information. Gel-free mass spectrometry proteomics techniques allow for rapid fingerprinting of bacterial proteins using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry or, for high-throughput sequencing of peptides from protease-digested cellular proteins, using mass analysis of fragments from collision-induced dissociation of peptide ions. The latter technique uses database searching of product ion mass spectra. A database contains a comprehensive list of protein sequences translated from protein-encoding open reading frames found in bacterial genomes. The results of such searches allow the assignment of experimental peptide sequences to matching theoretical bacterial proteomes. Phylogenetic profiles of sequenced peptides are then used to create a matrix of sequence-to-bacterium assignments, which are analyzed using numerical taxonomy tools. The results thereof reveal the relatedness between bacteria, and allow the taxonomic position of an investigated strain to be inferred. PMID- 16307517 TI - Biomarker discovery using protein microarray technology platforms: antibody antigen complex profiling. AB - Protein microarrays represent an important new tool in proteomic systems biology. This review focuses on the contributions of protein microarrays to the discovery of novel disease biomarkers through antibody-based assays. Of particular interest is the use of protein microarrays for immune response profiling, through which a disease-specific antibody repertoire may be defined. The antigens and antibodies revealed by these studies are useful for clinical assay development, with enormous potential to aid in diagnosis, prognosis, disease staging and treatment selection. The discovery and characterization of novel biomarkers specifically tailored to disease type and stage are expected to enable personalized medicine by facilitating preventative medicine, predictive diagnostics and individualized curative therapies. PMID- 16307518 TI - Emerging challenges in ligand discovery: new opportunities for chromatographic assay. AB - Ligand discovery initiatives are facing interesting challenges as ever-increasing numbers of proteins are entering screening programs. As an answer to steady pressure to improve performance in drug discovery, ligand discovery can expect to play an expanded role in generating small molecules as probes to help uncover the function of novel proteins. Chromatographic assay formats can offer new entry points into standard interaction characterization (binding and rate constants) as well as powerful, scaleable methods for compound screening. This review presents recent advancements in chromatographic assay technology, with a particular focus on frontal affinity chromatography as a platform technology for interaction analysis. PMID- 16307519 TI - How proteomics reveals potential biomarkers in brain diseases. AB - The brain is complex, and so are the proteomics studies of brain tissue and its diseases, including Alzheimer's Disease, Parkinson's Disease and schizophrenia. In this review, general considerations and strategies of proteomics technologies, the advantages and challenges as well as the special needs for brain tissue are described and summarized. In addition, the results of the first studies are presented including a quality evaluation of the candidate proteins for these diseases. A paragraph is dedicated to the efforts of standardization in this field. PMID- 16307520 TI - Improved understanding of gene expression regulation using systems biology. AB - This article reviews the current state of systems biology approaches, including the experimental tools used to generate 'omic' data and computational frameworks to interpret this data. Through illustrative examples, systems biology approaches to understand gene expression and gene expression regulation are discussed. Some of the challenges facing this field and the future opportunities in the systems biology era are highlighted. PMID- 16307521 TI - Proteomics of TGF-beta signaling and its impact on breast cancer. AB - The complexity of mechanisms leading to the appearance and progression of cancer is a challenge being addressed by large-scale studies, such as proteomics. Simultaneous monitoring of thousands of proteins uncovers novel signaling mechanisms, thus revising our knowledge of tumorigenesis. Transforming growth factor (TGF)-beta is a secreted polypeptide that is known to inhibit tumor growth at the early stages of cancer, but promote metastasis at the later stages. Proteomics-based studies have significantly widened our knowledge of TGF-beta dependent regulation of cell proliferation, apoptosis, DNA damage repair and transcription. This leads to better understanding of the TGF-beta role in human breast tumorigenesis, and opens the way for the development of novel anticancer treatments and drugs, with some of the drugs already entering clinics. This review discusses recent advances in proteomics studies of TGF-beta signaling and its contribution to the understanding and treatment of breast cancer. PMID- 16307522 TI - Microarray-based functional protein profiling using peptide nucleic acid-encoded libraries. AB - The availability of complete genome sequences from numerous organisms has provided investigators with the challenge of assigning physiologic functions to the encoded gene products. To facilitate this process, multiple technologies have been developed to profile the transcriptome and the proteome, including methods to monitor the function of enzymes in complex biologic systems. These methods typically target specific classes of enzymes and attempt to correlate the enzymatic activity with the specific phenotype of interest. Here, technologies to measure enzymatic activity on a subproteomic scale are reviewed, including the authors' own efforts, which are based on self-assembled microarrays utilizing peptide nucleic acid-encoded small-molecule libraries. PMID- 16307523 TI - Oxidative stress response: a proteomic view. AB - The oxidative stress response is characterized by various effects on a range of biologic molecules. When examined at the protein level, both expression levels and protein modifications are altered by oxidative stress. While these effects have been studied in the past by classic biochemical methods, the recent onset of proteomics methods has allowed the oxidative stress response to be studied on a much wider scale. The input of proteomics in the study of oxidative stress response and in the evidence of an oxidative stress component in biologic phenomena is reviewed in this paper. PMID- 16307524 TI - What is mzXML good for? AB - mzXML (extensible markup language) is one of the pioneering data formats for mass spectrometry-based proteomics data collection. It is an open data format that has benefited and evolved as a result of the input of many groups, and it continues to evolve. Due to its dynamic history, its structure, purpose and applicability have all changed with time, meaning that groups that have looked at the standard at different points during its evolution have differing impressions of the usefulness of mzXML. In discussing mzXML, it is important to understand what mzXML is not. First, mzXML does not capture the raw data. Second, mzXML is not sufficient for regulatory submission. Third, mzXML is not optimized for computation and, finally, mzXML does not capture the experiment design. In general, it is the authors' opinion that XML is not a panacea for bioinformatics or a substitute for good data representation, and groups that want to use mzXML (or other XML-based representations) directly for data storage or computation will encounter performance and scalability problems. With these limitations in mind, the authors conclude that mzXML is, nonetheless, an indispensable data exchange format for proteomics. PMID- 16307525 TI - Mechanism and consequences of delta-opioid receptor internalization. AB - G protein-coupled delta-opioid receptors (DORs) participate in opioid-mediated analgesia, and chronic opioid application is well known to produce tolerance, limiting the therapeutic use of these drugs. To control and eventually avoid the underlying adaptive mechanisms, several cellular functions were examined with regard to their roles in tolerance development. Specific interest focused on DOR internalization, and the relevant findings are reviewed here. In general, DOR endocytosis is accomplished by complex interactions of various determinants, each having distinct roles in this process. For instance, DOR activation by certain opioids has been shown to turn on the machinery of endocytosis, whereas other opioids stimulate the receptors but fail to bring about internalization. In addition, receptor phosphorylation by different kinases was commonly found to promote DOR sequestration, but receptor internalization also occurs without their phosphorylation. A central role in DOR endocytosis is referred to the adaptor proteins arrestin-2 and arrestin-3, which bind to receptors and subsequently cause the formation of clathrin-coated pits to trigger dynamin-controlled endocytosis. Distinct sorting proteins, kinases, and phosphatases determine whether internalized DORs are delivered either for proteolytic degradation or for recycling, although the underlying mechanisms are hence not clear. Despite intensive studies, understanding of DOR sequestration, degradation, and recycling becomes increasingly difficult. However, the phenomenon of cellular desensitization is recognized to correspond to the loss of responsiveness as consequence of DOR internalization and degradation. In contrast, DOR endocytosis is also discussed to promote resensitization of cells to opioids by recycling of internalized DORs. Even stimulation of extracellular signal-regulated protein kinases (ERK 1/2) may be accomplished by DOR sequestration. However, opposite findings, as well as the fact that multiple cellular mechanisms underly receptor desensitization, resensitization, and ERK activation, questions whether DOR internalization is essential for these processes. Further investigations in both the cellular mechanism and the consequences of DOR endocytosis might thus reveal new aspects of opioid-controlled functions. PMID- 16307526 TI - On the use of neuro-2a neuroblastoma cells versus intact neurons in primary culture for neurotoxicity studies. AB - Neuroblastoma cell lines have been used extensively to screen novel compounds for neurotoxic properties and associated mechanisms. Such transformed cell lines often display morphological, developmental, and signaling characteristics that are substantially different from the parental cell type. Consequently, the response of neuroblastoma cells to toxin exposure may differ from that of neurons. An appreciation of the pharmacological and functional differences between neurons and neuron-like cell lines is therefore essential when interpreting data derived from neuroblastoma-based assays. We have compared the effects of several neurotoxins on Ca2+ homeostasis and cell viability in cerebellar granule neurons (CGN) and a neuroblastoma cell line (Neuro-2a). To explore the mechanisms underlying differential sensitivity of intact neurons and neuroblastoma cells to neurotoxins, we also compared CGN and Neuro-2a cells for expression of voltage-gated sodium channels (VGSC) and N-methyl-D-aspartate receptors (NMDAR). Cytotoxic potency in neurons was several orders of magnitude greater for Caribbean-ciguatoxin-1 (C-CTX-1) than either domoate (Dom) or brevetoxin-2 (PbTx-2). In addition, the cytotoxic potency of C-CTX-1 was two orders of magnitude greater in CGN than in Neuro-2a cells. The effect of C-CTX-1 and Dom on calcium homeostasis was compared in fluo-3 loaded neurons. Dom caused an elevation in intracellular calcium ([Ca2+]i) at concentrations that paralleled the concentration/response relationship for cytotoxicity in CGN. Conversely, C CTX-1 did not elevate [Ca2+]i within the dynamic concentration range for cell death. The discordance of the concentration/response relationships for C-CTX-1 induced cytotoxicity and [Ca2+]i elevation suggests that acute C-CTX-1 cytotoxicity may involve mechanisms other than Ca2+ load. C-CTX-1-induced elevation of [Ca2+]i in neurons was dependent on activation of NMDAR and the reverse mode of operation of the Na+/Ca2+ exchanger. These data demonstrate that, although C-CTX-1, domoate, and PbTx-2 share the ability to produce neurotoxicity and mobilize calcium, their respective molecular targets and mechanisms of neurotoxicity differ. Neuro-2a cells that were not pretreated with veratridine and ouabain were insensitive to C-CTX-1 and glutamatergic agonists. VGSC expression was 20-fold lower in Neuro-2a cells than in CGN, whereas NMDARs were not expressed in these neuroblastoma cells. It is therefore likely that the enhanced sensitivity of CGN, relative to Neuro-2a cells, to neurotoxins is a consequence of pronounced differences in VGSC and NMDAR expression. These results underscore the need to exercise caution in interpreting negative cytotoxicity data derived from the use of neuroblastoma cell lines. PMID- 16307527 TI - Second-generation K(ATP) channel openers. AB - This review discusses structural aspects of second-generation K(ATP) channel openers (KCOs), which exhibit improved tissue-selectivity. Their therapeutic profile is debated with main focus on cardiac ischemia, asthma, and urinary incontinence. PMID- 16307528 TI - Natural products as angiogenesis modulators. AB - Cancer remains one of the major causes of death worldwide. Anti-angiogenic therapy is one of the new approaches to anticancer therapy. Out of 22 angiogenesis inhibitors currently under clinical trials there are 11 natural products or were modeled on a natural product parent. This review shows the potential of natural products for the discovery of new anti-angiogenic leads. PMID- 16307529 TI - Are vanadium compounds drugable? Structures and effects of antidiabetic vanadium compounds: a critical review. AB - Vanadate can be bioequivalent to phosphate and replace it in cellular metabolism. The detection of insulin-like activity has spurred interest in the development of oral anti-diabetic drugs containing vanadium. We collected and evaluated a vast toxicity data set and discussed molecular aspects related to insulin-mimetic effects of vanadium complexes. PMID- 16307530 TI - New fluoroquinolones: a class of potent antibiotics. AB - The discovery of antibiotics opened a new era in the treatment against several pathogenic microorganisms that can disable or kill humans. The appearance of the fluoroquinolones (based on nalidixic acid, 4-quinolone-3-carboxylates), in the early 1980's, gave a new impetus in the treatment of infectious diseases. Despite these favourable properties, the earlier fluoroquinolones had limited potency against some clinically important organisms, especially Gram-positive pathogens so that the development of resistance to these organisms has become a serious problem. Thus the development of new fluoroquinolones with a better pharmacokinetic profile, potency, broad spectrum of activity, solubility, prolonged serum half-life and oral and parenteral routes of administration has been a major focus on recent research. The increasing interest in this class led me to review the promising new fluoroquinolones in clinical trials. PMID- 16307531 TI - Non-insulin dependent diabetes mellitus: present therapies and new drug targets. AB - Type 2 Diabetes Mellitus (DM) or Non-Insulin Dependent Diabetes Mellitus (NIDDM) accounts for 90-95% of all diabetes cases and has become a major health concern over the years. This disease has assumed frightening proportions due to unhealthy food habits and sedentary life style. About a decade ago, due to the absence of defined molecular targets or an understanding of disease pathophysiology, treatment of this disease was mostly focused on insulin secretion or administration of external insulin. During the past decade however, advent of genomics and proteomics has helped in understanding the molecular alteration characteristics of NIDDM. Untreated type 2 diabetes leads to several complications such as hyperlipidemia, hypertension and atherosclerosis- collectively known as Syndrome X. Though United Kingdom Prospective Diabetes Study (UKPDS) showed that normalization of hyperglycemia could prevent majority of diabetes complications, the available treatment regime does not adequately normalize the blood glucose level in type 2 diabetic patients. Currently, four distinct classes of oral hypoglycemic agents are available, some of which can act as lipid lowering agents as well. The efficacy and side effect profiles of these drugs are still to be optimized, so there is an unmet need for better candidates. Several new targets as well as better drugs for old targets are under investigation across the world. Availability of such drugs, based on the validated targets, may lead to a new therapeutic paradigm for the prevention of diabetes as well as complications arising out of it. The current review will deal with existing oral therapies for type 2 diabetes as well as the emerging therapeutic targets. PMID- 16307532 TI - 1,2,4,5-Tetraoxacycloalkanes: synthesis and antimalarial activity. AB - In this short review the methods of preparation of novel 1,2,4,5 tetraoxacycloalkanes and the related peroxides are summarized, with the emphasis on the usefulness of 1,1-bishydroperoxides as the precursor. Also, their antimalarial activities in vitro and in vivo are discussed. PMID- 16307533 TI - Knowledge-driven lead discovery. AB - Virtual screening encompasses several computational approaches which have proven valuable for identifying novel leads. These approaches rely on available information. Herein, we review recent successful applications of virtual screening. The extension of virtual screening methodologies to target families is also briefly discussed. PMID- 16307534 TI - Quantitative mapping of oxidation-sensitive cysteine residues in SERCA in vivo and in vitro by HPLC-electrospray-tandem MS: selective protein oxidation during biological aging. AB - The selective reversible S-glutathiolation of specific SERCA (sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase) cysteine residues represents a novel physiologic pathway of NO (nitric oxide)-dependent arterial smooth muscle relaxation [Adachi, Weisbrod, Pimentel, Ying, Sharov, Schoneich and Cohen (2004) Nat. Med. 10, 1200-1207]. This mechanism may be impaired through the irreversible oxidation of functionally important cysteine residues as a consequence of oxidative stress and aging. To establish whether in vivo aging and in vitro oxidation by peroxynitrite result in the loss of such functionally important cysteine residues of SERCA, we have developed and optimized a quantitative method to monitor the oxidation state of the individual SERCA cysteine residues using a maleimide-based fluorescence dye, TG1 (ThioGlo 1), as a label for cysteine residues that have not been altered by oxidation and are not involved in disulphide bridges. A high efficiency for TG1 labelling of such residues and the chemical structure of cysteine-TG1 adducts were validated by MS analysis of model peptides, model proteins and rat skeletal muscle SERCA1. Tryptic peptides containing 18 out of a total of 24 cysteine residues were identified by HPLC-ESI (electrospray ionization)-MS/MS (tandem MS). Two cysteine residues, at positions 344 and 349, were detected in the form of an internal disulphide bridge, and another 16 were found to be labelled with TG1. Using HPLC-ESI-MS, we quantitatively mapped peroxynitrite oxidation of eight cysteine residues (positions 364, 417, 420, 498, 525, 674, 675 and 938), some of which are involved in the control of SERCA activity. Biological aging resulted in the partial modification of cysteine residues 377, 498, 525, 561, 614, 636, 674, 675, 774 and 938. Neither peroxynitrite exposure nor biological aging affected the apparent SERCA1 ATP affinity. Our data show an age-dependent loss of cysteine residues (approx. 2.8 mol of cysteine/mol of SERCA1), which may be partially responsible for the age-dependent decrease in the specific Ca2+-ATPase activity (by 40%). PMID- 16307535 TI - Digital imaging of haematological morphology. AB - Microscopic images of haematological cells are now routinely photographed using digital cameras. Advances in technology mean that the quality of such digital images can now approach that viewed through a microscope. At the same time there is an emerging appreciation that such images can be used in many roles: digital images are now being used to construct digital 'virtual slides', or are being employed together with cell recognition systems for morphological screening. Additionally, an Internet-based viewing systems allow access to on-line annotation, as well as real-time data gathering and feedback. The process of viewing digital images differs from the viewing of glass slides through a microscope; however, such images can provide diagnostic equivalence, and have an emerging role in areas such as education, quality control and continuing professional development. This review explores some of the present strengths, weaknesses and future applications of digital imaging in haematology. PMID- 16307536 TI - Slide session, British Society for Haematology, 45th Annual Scientific Meeting, Manchester, 2005. AB - Each year at the Annual Scientific Meeting of the British Society for Haematology, there is a slide session in which microscopic slides of six patients with haematological disorders are discussed by two experts. Further data and the final diagnosis are then provided. The slide session is presented here, as it occurred at the meeting. PMID- 16307537 TI - Platelet size has diagnostic predictive value in patients with thrombocytopenia. AB - We studied 473 unselected patients with thrombocytopenia. The mean platelet volume (MPV) was 8.1 fl in patients with marrow disease and 9.8 fl in patients without marrow disease (P < 0.001). A total of 5% of patients with an MPV >or=10.5 fl have marrow disease (odds ratio 0.05, 95% CI 0.02-0.13). Conversely over three quarters of patients with an MPV of <8.0 fl have marrow disease (odds ratio 8.1, 95% CI 5.0-13.0). Therefore the MPV can strongly guide the clinician as to the likely presence or absence of bone marrow disease in thrombocytopenic patients. PMID- 16307538 TI - A survey of the antibiotic treatment of febrile neutropenia in haematology units in the United Kingdom. AB - We performed a nationwide survey to define the different practices in managing febrile neutropenia in haematology units. A questionnaire was sent out to a named haematologist in each of 220 haematology units in the UK. Questions were asked regarding antibiotics of choice in managing febrile neutropenia and the use of antibiotic prophylaxis. Responses were received from 167 (76%) haematology units. Combination therapy with piperacillin-tazobactam and gentamicin is used first line in febrile neutropenia by 72% of units. Piperacillin-tazobactam monotherapy is used first-line by 5% of units. When response to initial empirical therapy does not occur after 24-48 h, 32% of haematology units add a glycopeptide (vancomycin or teicoplanin) and 31% change to a carbapenem and a glycopeptide. Seventy-one percent of units use oral fluoroquinolone prophylaxis for all neutropenic patients. The antibiotic treatment of febrile neutropenia in haematology patients, and the use of antibiotic prophylaxis, vary significantly across the UK. This survey is the first to examine the prescribing of UK haematology units in this area, and could help in the formulation of practice guidelines. PMID- 16307539 TI - Clinico-pathological study of Hodgkin's lymphoma in a cancer center in Taiwan. AB - The incidence rate of Hodgkin's lymphoma (HL) in Asia is much lower than that of western countries. This study demonstrates the incidence rate and the clinico pathological features of HL in a cancer center in Taiwan with respect to demographics, histological subtypes and clinical outcomes. We evaluated the clinical, morphological and immunohistochemical features of 42 patients with HL during the period of 1995-2002. Clinico-pathological features and follow-up were scrutinized. There were 21 males and 21 females. The incidence rate of HL in malignant lymphoma in our center was 7%. The median age was 26 years old. There was no apparent bimodal age distribution. The most prevalent histological subtype was nodular sclerosis (69%). Mixed cellularity, lymphocyte rich, lymphocyte depletion, nodular lymphocyte predominance and unclassified was 4.8%, 4.8%, 0%, 7% and 14% respectively. The most common site at presentation was the cervical lymph node (31 cases; 74%). Clinically, 1 (2%) had stage I disease, 23 (55%) stage II, 8 (19%) stage III and 10 (24%) stage IV. Two cases had rare primary bone marrow HL of stage IV. Both cases died within 1 month. Clinical stage (P=0.09) and age (P<0.001) were prognostic parameters determining the overall survival. PMID- 16307540 TI - Phenotype/genotype relationships in sickle cell disease: a pilot twin study. AB - The roles of genetic and non-genetic factors in the haematology, growth and clinical features of sickle cell disease have been studied in nine identical twin pairs (six homozygous sickle cell disease, three sickle cell-haemoglobin C disease). A comparison group of 350 age-gender matched sibling pairs, selected to have an age difference of <5 years, was used for assessing the concordance of numerical data. Attained height, weight at attained height, fetal haemoglobin, total haemoglobin, mean cell volume, mean cell haemoglobin and total bilirubin levels showed significantly greater correlation in identical twins than in siblings. Twins showed similarities in the prevalence and degree of splenomegaly, susceptibility to priapism, and in onset of menarche, but other clinical complications were discordant in prevalence and severity. These findings suggest that physical growth and many haematological characteristics are subject to genetic influences, but that non-genetic factors contribute to the variance in disease manifestations. PMID- 16307541 TI - Extramedullary haematopoiesis in the kidney: a case report and review of literature. AB - We report a 47-year-old man with myelofibrosis who presented with bilateral nephromegaly secondary to extramedullary haematopoiesis. We discuss diagnosis and treatment of this rare case and review the literature. PMID- 16307542 TI - Haemophagocytic syndrome associated with hepatitis-B virus infection responding to etoposide. AB - Haemophagocytic syndrome (HPS) secondary to viral infections usually has a variable course and can be life-threatening. We report a 53-year-old male patient who presented with fever, hepatosplenomegaly and pancytopenia. He had deranged liver function, abnormal clotting and markedly elevated serum ferritin. Bone marrow biopsy showed prominent haemophagocytosis. The patient was investigated thoroughly and found to have evidence of chronic hepatitis B-virus (HBV) infection by serological tests and liver biopsy. Other conditions associated with HPS such as lymphoma, malignancy and other viral or bacterial infections were not present. The patient did not respond to steroids, intravenous immunoglobulins or cyclosporin but responded to etoposide and became apyrexial. He also became HBV negative on lamivudine. The patient died of infection later on but there was no evidence of recurrence of HPS. To the best of our knowledge this is the first case report of HPS associated with isolated HBV infection. PMID- 16307543 TI - A case of all-trans retinoic acid-induced myositis in the treatment of acute promyelocytic leukaemia. AB - The use of all-trans retinoic acid (ATRA) is now standard therapy for the treatment of acute promyelocytic leukaemia (APML). There have been increasing reports of ATRA-induced myositis, with its frequent association with retinoic acid syndrome and Sweet's syndrome. We report a case of a young man with APML who developed ATRA-induced myositis characterized by unexplained fevers, bilateral leg swelling and a non-painful purpuric, petechial rash, with prompt resolution of symptoms and signs with high-dose steroids and cessation of ATRA. Rapid recognition of this adverse reaction and prompt institution of steroids is of prime importance given its potentially fatal course. PMID- 16307544 TI - Pyoderma gangrenosum as a cause of splenomegaly and association with a T-cell clone. AB - The spectrum of clinical presentation of haematological disease is wide. We highlight two features of this principle: a rare cause of a 'haematological' presentation and a possible haematological cause of a disease not normally considered as such. A case of systemic pyoderma gangrenosum presented with splenomegaly in the absence of a rash. A clonal gamma- and beta-T-cell receptor rearrangement was demonstrated. Such clones may be a general phenomenon involved in the pathogenesis of this condition. PMID- 16307545 TI - Successful treatment of a patient with chronic lymphocytic leukaemia (CLL) presenting with bony metastases with aggressive antibody and chemotherapy. AB - Osteolytic lesions are rare in chronic lymphocytic leukaemia (CLL) and thought to result from Richter's transformation or metastatic disease from nonlymphoid malignancies. We report a patient who presented with a large femoral metastatic lesion and hypercalcaemia caused by CLL itself. Complete remission of CLL with resolution of the osteolytic lesion was achieved with rituximab and cyclophosphamide, adriamycin, oncovin and prednisolone [CHOP (R-CHOP)] combination chemotherapy. PMID- 16307546 TI - Common origin of a rare beta-globin initiation codon mutation (ATG-->AGG) in Asians. AB - In this report, we describe two Thai siblings presenting with mild hypochromic microcytic anaemia and splenomegaly since 2(1/2) years of age. However, both patients were otherwise well with normal weight and height development and did not require transfusion during the 6-year follow-up period. Haematological and haemoglobin analyses were consistent with the clinical diagnosis of Hb E/beta thalassaemia disease. To provide proper genetic counselling for this family, a definitive diagnosis of beta-thalassaemia was achieved using molecular analysis. We identified a rare initiation codon mutation (ATG-->AGG) of the beta-globin gene in combination with the Hb E mutation (codon 26: GAG-->AAG). The initiation codon mutation has previously been reported in several East Asian populations but has never been found in Southeast Asia and in combination with Hb E before. The haplotype analysis revealed a common origin of this mutation in the Asian population (5': - + - + + - +: 3', type IV with framework 3 according to Orkin S, et al.). Although this rare mutation abolished the beta-globin expression and was considered as beta(0)-thalassaemia, the relatively mild phenotype in our patients may be attributed to a strong association between this mutation and the -158 (G)gamma (C-->T) polymorphism, an XmnI cleavage site (+), resulting in a high propensity of postnatal gamma-globin expression and ameliorating the clinical phenotypes. PMID- 16307547 TI - Clinical usefulness of transcription-mediated amplification and hybridization protection assay in imatinib-treated chronic myelogenous leukemia patients. PMID- 16307548 TI - Case clustering in infective endocarditis: the role of availability bias. AB - Limited data exist regarding the impact of variations in clinical practice and physicians' cognitive bias on the diagnosis of infective endocarditis (IE). As an illustration of these effects, unexpected clustering of IE diagnosis was encountered in a prospectively studied cohort. Transoesophageal echocardiography examinations for suspected IE were performed more frequently following a diagnosis of IE, and were associated with a subsequent cluster of IE cases. The cognitive bias of physicians resulting from a recent case of IE can lead to a transient increase in diagnosing additional cases of IE. PMID- 16307549 TI - Probiotics: facts and myths. AB - In recent years there has been a significant upsurge in research on the characterisation and verification of the potential health benefits associated with the use of probiotics. In addition, the market for probiotics continues to expand exponentially as consumers (mostly healthy individuals) rely on health claims made by manufacturers to make their choices. This review appraises the available evidence for and against the health claims associated with probiotics. The use of probiotics in promoting gastrointestinal health and immunity, and their use in the prevention of urogenital infections, allergies and cancer are reviewed. Furthermore, issues surrounding the use of probiotics in healthy individuals, the safety of probiotics and regulatory concerns are addressed. There is scientific evidence that specific strains of probiotic microorganisms confer health benefits on the host and are safe for human use. However, this evidence cannot be extrapolated to other strains, as these effects are strain specific. Probiotics have potential health benefits for conditions such as gastrointestinal infections, genitourinary infections, allergies and certain bowel disorders, all of which afflict a considerable proportion of the global population. However, considerable work is still needed to confirm these potential health benefits. PMID- 16307550 TI - The genetics of staphylococcal biofilm formation--will a greater understanding of pathogenesis lead to better management of device-related infection? AB - Staphylococcus epidermidis and Staphylococcus aureus are common causes of biofilm mediated prosthetic device-related infection. The polysaccharide adhesion mechanism encoded by the ica operon is currently the best understood mediator of biofilm development, and represents an important virulence determinant. More recently, the contributions of other virulence regulators, including the global regulators agr, sarA and sigmaB, to the biofilm phenotype have also been investigated. Nevertheless, little has changed at the bedside; the clinical and laboratory diagnosis of device-related infection can be difficult, and biofilm resistance frequently results in failure of therapy. This review assesses the way in which advances in the understanding of biofilm genetics may impact on the clinical management of device-related infection. PMID- 16307551 TI - Antimicrobial activity of doripenem (S-4661): a global surveillance report (2003). AB - The spectrum of activity and potency of doripenem, a broad-spectrum parenteral carbapenem currently in clinical development, was evaluated using 16 008 clinical bacterial isolates collected as part of an international surveillance project during 2003. Using reference broth microdilution methods, doripenem was found to be highly active against oxacillin-susceptible Staphylococcus aureus and coagulase-negative staphylococci (2705 and 297 isolates, respectively; MIC90s 0.06 mg/L), with a potency greater than that of other carbapenem antibiotics. Against enterococci (1474 isolates), with the exception of Enterococcus faecium, doripenem displayed modest activity (MIC50 4). Doripenem was among the most potent agents tested against Streptococcus pneumoniae, viridans group streptococci and beta-haemolytic streptococci (885, 140 and 397 isolates; MIC(90)s 0.5, 0.5 and 0.03 mg/L, respectively). For Enterobacteriaceae (> 6200 isolates), doripenem was four- to 32-fold more active than imipenem against wild type isolates (MIC90s 0.03-0.5 mg/L). MIC90s for confirmed extended-spectrum beta lactamase-producing Escherichia coli and Klebsiella pneumoniae (121 and 155 isolates; 0.06 and 0.12 mg/L, respectively) were two-fold higher than for wild type isolates. Doripenem was also active against Citrobacter spp., Enterobacter spp. and Serratia spp. (MIC90s 0.06-0.25 mg/L), including ceftazidime-resistant isolates. Doripenem and meropenem were the most active agents among all beta lactams against Pseudomonas aeruginosa (829 isolates; MIC50/90s 0.5/8 and 0.5/16 mg/L, respectively), whereas doripenem and imipenem were the most active agents against Acinetobacter spp. (155 isolates; MIC50/90s 0.5/4 and or= 65 years. In three university hospitals, blood cultures were obtained for only 37% of patients with community-acquired pneumonia, and fewer than one-third of such cultures were obtained in one hospital before antibiotics were prescribed, suggesting that the true incidence of IPD was closer to 50/100 000. PMID- 16307553 TI - Predicted effects on antibiotic use following the introduction of British or North American guidelines for community-acquired pneumonia in The Netherlands. AB - This study evaluated the possible changes in antibiotic use that might follow the implementation of British or North American guidelines for the treatment of community-acquired pneumonia (CAP) in The Netherlands. Patients admitted for mild, moderate and severe CAP were evaluated prospectively. Volume of antibiotic use, based upon guidelines of the British Thoracic Society (BTS), the Infectious Diseases Society of America (IDSA) or the American Thoracic Society (ATS), was estimated and compared to current practice. For 248 patients, current antibiotic use was 3087 defined daily doses. Antibiotic use would increase by 38% if based on ATS guidelines, by 23% if based on IDSA guidelines, and by 21% if based on BTS guidelines. The most significant increase in antibiotic use would occur for cases of moderate CAP, with incremental antibiotic costs of 1 750 000-3 500 000 Euros in The Netherlands. PMID- 16307554 TI - Efficacy of high loading doses of liposomal amphotericin B in the treatment of experimental invasive pulmonary aspergillosis. AB - This study aimed to investigate whether initial treatment of experimental pulmonary aspergillosis with high loading doses can be used as an alternative to standard therapeutic regimens. Steroid-immunosuppressed rats, infected intratracheally with Aspergillus fumigatus, received either amphotericin B deoxycholate (d-AmB) 1 mg/kg/day, liposomal amphotericin B (L-AmB) 5 mg/kg/day, or underwent a 3-day course of L-AmB 10 mg/kg, or 10 mg/kg for the first 3 or 4 days of treatment, followed by 3 mg/kg until the end of treatment. Therapy started 24 h after fungal challenge and lasted for 7 days. Compared to controls, survival was improved significantly in animals receiving any L-AmB regimen (p 70% since implementation of this protocol, while annual vancomycin utilisation has shown only small, incremental increases. Therefore, it is suggested that routine AST of CNS isolates from blood culture is not essential. PMID- 16307562 TI - Activity of five quinolones, three macrolides and telithromycin against 12 Haemophilus influenzae strains with different resistance phenotypes. AB - Gemifloxacin MICs for 12 Haemophilus influenzae strains with different resistance phenotypes were 0.001-0.015 mg/L. Gemifloxacin was bactericidal against all 12 strains after 24 h at 2 x MIC. Ciprofloxacin, levofloxacin, gatifloxacin and moxifloxacin had MICs of 0.008-0.03 mg/L and similar kill kinetics. Macrolides and telithromycin had unimodal MICs (1.0-8.0 mg/L), except for two strains without efflux systems (0.0125-0.5 mg/L) and two with efflux systems and ribosomal protein mutations (> 64.0 mg/L), and were bactericidal against eight to ten strains tested at 2 x MIC after 24 h. PMID- 16307563 TI - Analysis of the mechanisms of resistance to several antimicrobial agents in Shigella spp. causing travellers' diarrhoea. AB - Eighty isolates of Shigella spp. (37 Shigella flexneri and 43 Shigella sonnei) from patients with travellers' diarrhoea were studied. Susceptibility tests revealed high levels of resistance, especially to ampicillin (65%), tetracycline (78%) and trimethoprim (75%), and particularly among the S. flexneri isolates. Dihydrofolate reductase 1 genes (dfrA1) were prevalent among the trimethoprim resistant isolates, while oxa genes predominated among the ampicillin-resistant isolates. Chloramphenicol resistance was associated with production of chloramphenicol acetyltransferase, while nalidixic acid-resistant isolates had a single mutation in the gyrA gene. The results indicate a continuing need for resistance surveillance and rational use of antimicrobial agents. PMID- 16307564 TI - Bactericidal activity of fluoroquinolones against plasmid-mediated QnrA-producing Escherichia coli. PMID- 16307565 TI - Correct use of the term 'pan-drug-resistant' (PDR) Gram-negative bacteria. PMID- 16307566 TI - Serum procalcitonin levels in patients with mild community-acquired pneumonia. PMID- 16307567 TI - Elevated serum transaminases in patients with Mycoplasma pneumoniae pneumonia. PMID- 16307568 TI - Early loading of non-submerged titanium implants with a sandblasted and acid etched surface. 5-year results of a prospective study in partially edentulous patients. AB - OBJECTIVES: The aim of this prospective cohort study was to evaluate the success rate of titanium screw-type implants with the sandblasted and acid-etched (SLA) surface loaded early, after 6 weeks of healing. MATERIAL AND METHODS: A total of 104 implants were inserted into posterior sites of 51 partially edentulous patients exhibiting bone densities of class I-III. After a healing period of 6 weeks, all implants were functionally loaded with cemented crowns or fixed partial dentures. The patients were recalled at 3, 12, 24, 36, 48 and 60 months for clinical and radiographic examination. RESULTS: One implant failed to integrate during healing, and three implants were lost to follow-up and were considered drop-outs. The remaining 100 implants showed favorable clinical and radiographic findings at the 5-year examination. The peri-implant soft tissues were stable over time; the mean probing depths and mean attachment levels did not change during the follow-up period. None of the radiographs exhibited signs of continuous peri-implant radiolucency, which confirmed ankylotic stability for all 100 implants. The measurement of the bone crest levels (DIB values) indicated stability as well. Based on strict success criteria, all 100 implants were considered successfully integrated, resulting in a 5-year success rate of 99%. CONCLUSION: This prospective study using an early loading protocol with 6 weeks of healing demonstrated that titanium implants with the SLA surface can achieve and maintain successful tissue integration with high predictability for at least 5 years of follow-up in selected patients and sites. PMID- 16307569 TI - Evaluation of soft tissue around single-tooth implant crowns: the pink esthetic score. AB - AIM: In this study, the reproducibility of a newly developed pink esthetic score (PES) for evaluating soft tissue around single-tooth implant crowns was assessed. The effect of observer specialization was another point of interest. MATERIAL AND METHODS: Twenty observers (five prosthodontists, five oral surgeons, five orthodontists and five dental students) were given photographs of 30 single-tooth implant crowns. Seven variables were evaluated vs. a natural reference tooth: mesial papilla, distal papilla, soft-tissue level, soft-tissue contour, alveolar process deficiency, soft-tissue color and texture. Using a 0-1-2 scoring system, 0 being the lowest, 2 being the highest value, the maximum achievable PES was 14. Each observer was requested to make two assessments at an interval of 4 weeks. At the second assessment, the photographs were scored in the reverse order. RESULTS: The mean PES of evaluations at the first assessment (n=600) was 9.46 (+/-3.81 SD), and 9.24 (+/-3.8 SD) at the second one. The difference between these two means was not significant statistically (P=0.6379). Implant-related mean PES for single-tooth implants varied from 2.28 to 13.8, with standard deviations between 0.46 and 3.51. Very poor and very esthetic restorations showed the smallest standard deviations. The mean total PES was 10.6 for the prosthodontists, 9.2 for the oral surgeons, 9.9 for the dental students and 7.6 for the orthodontists. CONCLUSIONS: The PES reproducibly evaluates peri-implant soft tissue around single-tooth implants. Thus, an objective outcome of different surgical or prosthodontic protocols can be assessed. Orthodontists were clearly more critical than the other observers. PMID- 16307570 TI - A new index for rating aesthetics of implant-supported single crowns and adjacent soft tissues--the Implant Crown Aesthetic Index. AB - OBJECTIVES: The important item of aesthetics is rarely included in evaluation studies. The aim of this study was to develop and validate an index for rating aesthetics of implant-supported single crowns and adjacent soft tissues. MATERIAL AND METHODS: Nine items were selected, which have an influence on the aesthetic result. The items are based on the anatomic form, colour and surface characteristics of the crown and on the anatomic form, colour and surface characteristics of the peri-implant soft tissues. Two oral-maxillofacial surgeons and two prosthodontists rated 24 implant-supported single-tooth restorations and adjacent soft tissues on a form with the nine items of the rating index. The rating was carried out twice by each of the examiners. Weighted Cohen's kappa was calculated to express the intra- and interobserver agreement. RESULTS: Intraobserver results indicated that the agreement between the first and second rating of both the prosthodontists was good (both 0.7) and that the agreement of the oral-maxillofacial surgeons was moderate (0.49 and 0.56). The best interobserver agreement was found between the two prosthodontists (0.61, good agreement). CONCLUSIONS: The Implant Crown Aesthetic Index is an objective tool in rating aesthetics of implant-supported single crowns and adjacent soft tissues. The rating is best be carried out by one prosthodontist to have the highest reliability. PMID- 16307571 TI - Effects of implant surface microtopography on osteoblast gene expression. AB - AIM: The promotion of osteoblast attachment and differentiation has been evaluated on various implant surfaces. However, the effects of different implant surface properties on gene expression of key osteogenic factors are not fully understood. OBJECTIVE: The objectives of this study were to evaluate how topographical effects on titanium surface alter the expression of bone-related genes and transcription factors. METHODS: Osteoblasts were cultured on titanium disks prepared with a titanium dioxide grit blasting (TiOBlast) or grit blasted and etched with hydrofluoric acid (Osseospeed), grit blasted and etched (SLA-1), or grit blasted, etched and rinsed with N2 protection and stored in isotonic NaCl (SLA-2) commercially pure titanium implant discs. High-density cultures of human mesenchymal pre-osteoblastic cells (HEPM 1486, ATCC) were grown for 72 h and real time PCR used for quantitative analysis of alkaline phosphatase (ALP), core binding factor alpha1 (Cbfa1), Osterix, Type I Collagen, Osteocalcin and bone sialoprotein II gene expression. RESULTS: Real-time PCR showed significant (P<0.001) increases in ALP gene expression in osteoblasts grown on SLA-2, relative to all other surfaces. Cbfa1/RUNX-2 gene expression was significantly (P<0.01) increased on Osseospeed and TiOBlast surface as compared with SLA-1 and SLA-2 surfaces. The expression of Osterix had a trend similar to that of Cbfa1. CONCLUSION: In conclusion, implant surface properties may contribute to the regulation of osteoblast differentiation by influencing the level of bone-related genes and transcription factors in human mesenchymal pre-osteoblastic cells. PMID- 16307572 TI - Comparison of osteoblast spreading on microstructured dental implant surfaces and cell behaviour in an explant model of osseointegration. A scanning electron microscopic study. AB - OBJECTIVES: To compare interactions between rat calvarial osteoblasts and titanium dental implants with different microstructured surfaces. MATERIAL AND METHODS: Seven commercially available implants were used. Surfaces included plasma-sprayed, grit-blasted and/or acid-etched, smooth-machined and anodised titanium. Two methods were used to compare cell behaviour: (1) A cell-spreading assay in which percentages of cells at four different stages of attachment were identified by scanning electron microscopy and quantified within a 30 min attachment period. (2) Implants were placed in 'pocket culture' within nylon mesh sacs in contact with explanted calvarial bone fragments for 2 and 4 weeks. RESULTS: Surfaces combining grit blasting and acid etching, of microporous topography, showed significantly enhanced rates of cell spreading in comparison with the others. Differential cell morphology was observed in both suspension assays and pocket cultures. In the latter, cells migrated onto all surfaces. Multicellular layers with extracellular matrix (ECM) were present between the layers and on the material surfaces after 2 weeks. After 4 weeks, cell layers were more consolidated, and microstructures were obscured by layers of cells and ECM. Mineralised tissue was seen in association with ECM on grit-blasted surfaces of rough and smooth microtopography. CONCLUSIONS: The two methods provided complementary information: a rough surface of porous microstructure may enhance the rate of cell spreading. Differentiation and calcification occurred on surfaces of both rough and smooth microstructure. PMID- 16307573 TI - Effect of three distinct treatments of titanium surface on osteoblast attachment, proliferation, and differentiation. AB - Cell-titanium interactions are crucial to the clinical success of bone and dental implants. The physico-chemical characteristics of the substrates surface influence osteoblast proliferation, differentiation, and activity as well. The osteoblast behavior was analyzed on three different titanium surfaces: ground with an abrasive 600 grit SiC paper, blasted with alumina particles (65 microm diameter) and alumina blasted followed by a double chemical etch (4% HF+4% HF/8% H2O2). Scanning electron microscopy (SEM) and profilometry showed distinct microtopographies. Ground samples showed parallel-groove orientation. The Al2O3 blasted surface presented the roughest microtopography with aluminum-rich particles incrusted in the titanium surface. Osteoblasts cells from femora of Balb/c mice were seeded onto the substrates tested. Cell morphology and initial attachment were evaluated by SEM. Osteoblasts adhered to and spread on all samples tested. However, on rough surfaces, osteoblasts did not spread completely and acquired a polygonal morphology. Besides, the cell proliferation rate was diminished at the beginning of incubation on rough surfaces. Our results suggest a delay, rather than an impairment, in osteoblast viability and alkaline phosphatase activity when cells are cultured on rough surfaces, inducing a distinct osteoblast phenotype, rather than blocking its activity. At least in the culture conditions used in this work, alumina particles did not affect osteoblast behavior. PMID- 16307574 TI - Platelet-rich plasma and fibrin as delivery systems for recombinant human bone morphogenetic protein-2. AB - The aim of the present study was (1) to test whether or not platelet-rich plasma (PRP) or commercially available fibrin can increase bone regeneration compared with non-treated defects and (2) to test whether or not PRP or fibrin increases bone regeneration when used as a delivery system for recombinant human bone morphogenetic protein-2 (rhBMP-2). In 16 New Zealand White rabbits, four evenly distributed 6 mm diameter defects were drilled into the calvarial bone. The following five treatment modalities were randomly allocated to all 64 defects: (0) untreated control, (1) fibrin alone, (2) PRP alone, (3) fibrin with 15 microg rhBMP-2 and (4) PRP with 15 microg rhBMP-2. For the fibrin gels and the PRP containing rhBMP-2, the 15 microg rhBMP-2 was incorporated by precipitation within the matrices before their gelation. After 4 weeks, the animals were sacrificed and the calvarial bones were removed for histological preparation. The area fraction of newly formed bone was determined in vertical sections from the middle of the defect by applying histomorphometrical analysis. A mean area fraction of newly formed bone was found within the former defect of 23.4% (+/ 13.5%) in the control sites, of 28.4% (+/-17.4%) in the fibrin sites and of 34.5% (+/-17.4%) in the PRP sites. The statistical analysis revealed no significant difference in bone formation between the three groups (ANOVA). Addition of 15 microg rhBMP-2 in the fibrin gel (59.9+/-20.3%) and the PRP gels (63.1+/-25.3%) increased bone formation significantly. No significant difference was observed between sites, where PRP or fibrin has been used as a delivery system for rhBMP-2 (ANOVA). In conclusion, the application of fibrin gels or PRP gels to bone defects is not superior to leaving the defect untreated. Regarding the amount of bone formation, the application of 15 microg rhBMP-2 in bone defects enhances the healing significantly at 4 weeks. In this animal model, commercially available fibrin and autologous PRP gels are equally effective as delivery systems for rhBMP-2. PMID- 16307575 TI - Bone healing in surgically created defects treated with either bioactive glass particles, a calcium sulfate barrier, or a combination of both materials. A histological and histometric study in rat tibias. AB - OBJECTIVE: The purpose of this study was to histologically analyze the influence of bioactive glass and/or a calcium sulfate barrier on bone healing in surgically created defects in rat tibias. MATERIAL AND METHODS: Sixty-four rats were divided into 4 groups: C (control), CS (calcium sulfate), BG (bioactive glass), and BG/CS (bioactive glass/calcium sulfate). A surgical defect was created in the tibia of each animal. In Group CS, a calcium sulfate barrier was placed to cover the defect. In Group BG the defect was filled with bioactive glass. In Group BG/CS, it was filled with bioactive glass and protected by a barrier of calcium sulfate. Animals were sacrificed at 10 or 30 days post-operative. The formation of new bone in the cortical area of the defect was evaluated histomorphometrically. RESULTS: At 10 days post-operative, Group C presented significantly more bone formation than Groups CS, BG, or BG/CS. No statistically significant differences were found between the experimental groups. At 30 days post-operative, Group C demonstrated significantly more bone formation than the experimental groups. Groups CS and BG/CS showed significantly more bone formation than Group BG. No statistically significant differences were found between Group CS and BG/CS. CONCLUSIONS: (a) the control groups had significantly more bone formation than the experimental groups; (b) at 10 days post-operative, no significant differences were found between any of the experimental groups; and (c) at 30 days post-operative, the groups with a calcium sulfate barrier had significantly more bone formation than the group that used bioactive glass only. PMID- 16307576 TI - Osteotome sinus elevation and simultaneous placement of porous-surfaced dental implants: a morphometric study in rabbits. AB - The objective was to establish a model in rabbits in which to study the healing events associated with localized indirect osteotome-mediated maxillary sinus floor elevation in conjunction with simultaneous placement of sintered porous surfaced dental implants. On one side of the maxilla of each of 28 rabbits, a sintered porous-surfaced titanium alloy press-fit implant was placed without the use of a bone graft material, while on the collateral side an implant was placed after first adding Bio-Oss graft particles to the osteotomy. Specimens were retrieved for morphometric assessment of bone contact and bone ingrowth of the porous implant surface after 2, 4, 6 and 8 weeks of healing. All implants became osseointegrated by bone ingrowth into the porous implant surface. While the addition of graft particles did not result in a statistically significant increase in the parameters measured, a trend for greater bone contact and particularly bone ingrowth at the apices of the implants was seen as healing time increased. The rabbit maxillary sinus can be used to study healing following placement of sintered porous-surfaced dental implants using the indirect sinus elevation procedure. PMID- 16307577 TI - Bi-directional distraction osteogenesis of the alveolar bone using an extraosseous device. AB - OBJECTIVES: In alveolar distraction, the bone segment tends to incline palatally or lingually, making rigid control of the segments difficult. The aim of this study was to evaluate the usefulness of a newly developed bi-directional extraosseous alveolar distractor (Medartis V2-Alveolar distractor) for pre- and perioperative vector management. MATERIAL AND METHODS: Seven patients with segmental alveolar atrophy following traumatic tooth loss were treated using the distraction device. The patients were followed up clinically and radiologically. Preoperatively, the initial vector for distraction was determined using CT by measuring the cross-section of the bone. The morphology of the alveolar bone was also analyzed in relation to the planned implant position. Postoperatively, the rate of osteogenesis was monitored with plane radiographs and CT scan. RESULTS: All cases had bone deficit at the anterior surface of the alveolar ridge, showing a typical inclination of the long axis of the bone. Using the distractor, vertical distraction and positioning of the segments with labial orientation was possible. After a consolidation period of 12 weeks on average, sufficient bone formation for implant installation was radiologically observable. Histologic and histomorphometric analysis of one bone biopsy showed very dense mineralized bone (area fraction=78%) with a multidirectional, complex architecture. Implant supported prosthetic oral rehabilitation was successfully performed in all cases. CONCLUSION: All complications observed in this study were related to the bone deficiency at the anterior surface of the alveolar process. If the technique can be improved, this type of bi-directional distraction is a promising method for alveolar bone repair. PMID- 16307578 TI - Histomorphometry and micro-computed tomography of bone augmentation under a titanium membrane. AB - OBJECTIVES: Bone augmentation underneath an occlusive titanium membrane is evaluated in most cases by means of serial histological sections and histomorphometry. Micro-computed tomography (micro-CT) is a less invasive and dynamic technique to measure bone volume in animals of a size that fits into the gantry. The aim of the present study was to evaluate whether the latter approach could match histomorphometry to assess bone augmentation under a titanium membrane. MATERIAL AND METHODS: Pre-formed titanium cups were placed on the skull of 16 rabbits. Bone formation underneath the cups was allowed to occur for 12 weeks. The amount of bone volume assessed by micro-CT was expressed as a numerical unit. One unit volume corresponds to 0.043 mm3. The measurements reveal the volume of bone-like tissue under the membrane, with the same density as that of the original rabbit skull bone. Histological sections were cut along the same plane as the one used for the micro-CT images. The total bone surface was assessed by a digital image system in double-stained undecalcified histological sections and related to the maximum available surface of the titanium cups, which was on average 1366 mm2. RESULTS: The amount of total bone surface found under the titanium membrane varied between 40 and 163 mm2. Measured by micro-CT, the bone detected ranged from 3.7 to 396 numerical units. A highly significant (P<0.001) correlation was found between the total bone volume measured in conventional serial histological sections and by the micro-CT technique (r2=0.72). CONCLUSIONS: The total bone volume measured underneath a membrane using the micro-CT when compared with histological sections remained within a 16% error. This is because of the scattering effect of the metallic membrane and the impossibility to distinguish newly formed bone from the original skull bone on the micro-CT images. PMID- 16307579 TI - Human ex vivo bone tissue strains around immediately loaded implants supporting maxillary overdentures. AB - PURPOSE: To evaluate ex vivo bone tissue strains around maxillary implants supporting immediately loaded bar-retained overdentures. MATERIAL AND METHODS: Ninety degree two-element rosette strain gauges were bonded on the labial bone of four ITI dental implants placed in the maxillary anterior region of four completely edentulous maxilla of fresh human cadavers. The installation torque value (ITV) of each implant was measured by a custom-made torque wrench and resonance frequency analyses (RFAs) were undertaken. A bar-retained overdenture was fabricated for each cadaver, and two miniature load cells were integrated in the first molar region of the overdentures for controlled loading experiments. Strain measurements were performed at a sample rate of 10 kHz and under a maximum load of 100 N, simultaneously monitored from a computer connected to a data acquisition system. Finally, removal torque values (RTVs) of the implants were measured. RESULTS: RFA values did not mirror ITVs, while RTVs of implants were slightly lower than the ITVs. Any correlation could not be obtained between RFA values and ITVs or RTVs. Maximum strains around loaded implants ranged between 100 and -550 microepsilon under 25-100 N. The axial and lateral strain values of posterior implants of both sides were higher than those of anterior implants under all loads (P<0.05). CONCLUSION: Because occlusal forces in humans tend to decrease because of age-related factors, maximum strains around immediately loaded implants supporting maxillary overdentures fall within physiologic levels. PMID- 16307580 TI - Rehabilitation with dental prosthesis can increase cerebral regional blood volume. AB - Treatment with denture for edentulous people is highly important for maintaining quality of life. However, its effect on the brain is unknown. In this experimental study, we hypothesized that dental prosthesis can recover not only the physical condition of mastication system but also the regional brain activity. We evaluated functional brain imaging of edentulous subjects fixed by dental implant prosthesis with clenching tasks by multi-channel near-infrared optical topography. Results revealed a significantly (P<0.001; paired t-test) increased cerebral regional blood volume during maximum voluntary clenching task by implant-retained prosthesis. There were no statistically significant differences between patients with and without prosthesis in the latency to the maximum regional blood volume after the task. Conclusively, clenching can be effective for increasing cerebral blood volume; accordingly maintenance of normal chewing might prevent the brain from degenerating. PMID- 16307581 TI - A novel method of removing artifacts because of metallic dental restorations in 3 D CT images of jaw bone. AB - CT images, especially in a three-dimensional (3-D) mode, give valuable information for oral implant surgery. However, image quality is often severely compromised by artifacts originating from metallic dental restorations, and an effective solution for artifacts is being sought. This study attempts to substitute the damaged areas of the jaw bone images with dental cast model images obtained by CT. The position of the dental cast images was registered to that of the jaw bone images using a devised interface that is composed of an occlusal bite made of self-curing acrylic resin and a marker plate made of gypsum. The patient adapted this interface, and CT images of the stomatognathic system were filmed. On the other hand, this interface was placed between the upper and lower cast models and filmed by CT together with the cast models. The position of the marker plate imaged with the dental casts was registered to those adapted by the patient. The error of registration was examined to be 0.25 mm, which was satisfactory for clinical application. The damaged region in the cranial bone images as an obstacle for implant surgery was removed and substituted with the trimmed images of the dental cast. In the method developed here, the images around the metallic compounds severely damaged by artifacts were successfully reconstructed, and the stomatognathic system images became clear, and this is useful for implant surgery. PMID- 16307582 TI - Adult neurogenesis in the central olfactory pathway in the absence of receptor neuron turnover in Libinia emarginata. AB - Life-long neurogenesis is a characteristic feature of the olfactory pathways of a phylogenetically diverse array of animals. In both vertebrates and invertebrates, the life-long addition of olfactory interneurons in the brain occurs in parallel with the continuous proliferation of olfactory receptor neurons in the olfactory organ. It has been proposed that these two processes are related functionally, with new olfactory interneurons being added to accommodate the new olfactory receptor neurons added in the periphery. While this has not been tested directly because the two processes are not readily separable, this question can be addressed in the olfactory pathway of the crab, Libinia emarginata. Unlike most decapod crustaceans, which moult and grow throughout life, L. emarginata has a terminal, maturational moult after which animals become anecdysic (stop moulting). Because the addition of new receptor neurons in crustaceans is associated with moulting, a comparison of neurogenesis in immature and mature L. emarginata provides an opportunity to examine the interdependence of central and peripheral neurogenesis in the olfactory pathway. This study demonstrates that the continuous addition of olfactory receptor neurons in L. emarginata ceases at the terminal moult but that proliferation and differentiation of olfactory interneurons in the brain continues in mature animals. Contrary to the general assumption, therefore, continuous neurogenesis in the central olfactory pathway of this species does not occur as part of a process involving the coregulation of central and peripheral neurogenesis. These findings suggest that peripheral neurogenesis is not a requirement for continuous neurogenesis in the central olfactory pathway. PMID- 16307583 TI - Ret deficiency in mice impairs the development of A5 and A6 neurons and the functional maturation of the respiratory rhythm. AB - Although a normal respiratory rhythm is vital at birth, little is known about the genetic factors controlling the prenatal maturation of the respiratory network in mammals. In Phox2a mutant mice, which do not express A6 neurons, we previously hypothesized that the release of endogenous norepinephrine by A6 neurons is required for a normal respiratory rhythm to occur at birth. Here we investigated the role of the Ret gene, which encodes a transmembrane tyrosine kinase receptor, in the maturation of norepinephrine and respiratory systems. As Ret-null mutants (Ret-/-) did not survive after birth, our experiments were performed in wild-type (wt) and Ret-/- fetuses exteriorized from pregnant heterozygous mice at gestational day 18. First, in wt fetuses, quantitative in situ hybridization revealed high levels of Ret transcripts in the pontine A5 and A6 areas. Second, in Ret-/- fetuses, high-pressure liquid chromatography showed significantly reduced norepinephrine contents in the pons but not the medulla. Third, tyrosine hydroxylase immunocytochemistry revealed a significantly reduced number of pontine A5 and A6 neurons but not medullary norepinephrine neurons in Ret-/- fetuses. Finally, electrophysiological and pharmacological experiments performed on brainstem 'en bloc' preparations demonstrated impaired resting respiratory activity and abnormal responses to central hypoxia and norepinephrine application in Ret-/- fetuses. To conclude, our results show that Ret gene contributes to the prenatal maturation of A6 and A5 neurons and respiratory system. They support the hypothesis that the normal maturation of the respiratory network requires afferent activity corresponding to the A6 excitatory and A5 inhibitory input balance. PMID- 16307584 TI - Ephrin-A4 inhibits sensory neurite outgrowth and is regulated by neonatal skin wounding. AB - The mechanisms for directing and organising sensory axons within developing skin remain largely unknown. The present study provides the first evidence that signalling occurs between A-ephrins and Eph-A receptors during the development of rat cutaneous sensory innervation both during normal development and following skin injury. Specifically, our data indicate that ephrin-A4 mRNA and protein are expressed in the epidermis during late embryogenesis and the early postnatal period (E16-P3), and expression is significantly down-regulated postnatally. In addition, Eph-A receptors are expressed on dorsal root ganglia (DRG) cells at birth. The pattern of ephrin-A4 expression is mirrored by epidermal innervation, so that sensory terminals are restricted to epidermal regions devoid of ephrin-A4 but increase as ephrin-A4 expression subsides postnatally. Neonatal skin wounding causes sensory hyperinnervation and a differential screen of wounded vs. nonwounded skin revealed down-regulation of epidermal ephrin-A4 following neonatal skin wounding. Expression studies showed that this down-regulation is below the wound and coincides exactly with the onset of hyperinnervation. In vitro experiments show a function for ephrin-A4-Fc in inhibiting rat DRG neuronal growth and guidance when presented as either substratum-bound stripes of ephrin A4-Fc or as soluble clustered proteins. In conclusion, these observations suggest that the Eph family ligand ephrin-A4 has an inhibitory influence on neonatal cutaneous nerve terminals from DRG sensory neurons in the hindlimb, and may serve to prevent inappropriate innervation of cutaneous regions. In addition, the absence of ephrin-A4 following neonatal skin wounding may play a critical permissive role in the sprouting response. PMID- 16307585 TI - Dopamine D3 receptor-preferring agonists induce neurotrophic effects on mesencephalic dopamine neurons. AB - Anti-parkinsonian agents, pramipexole (PPX) and ropinirole (ROP), have been reported to possess neuroprotective properties, both in vitro and in vivo. The mechanisms underlying neuroprotection afforded by the D3-preferring receptor agonists remain poorly understood. The present study demonstrates that incubation of primary mesencephalic cultures with PPX and ROP or the conditioned medium from PPX- or ROP-treated primary cultures induced a marked increase in the number of dopamine (DA) neurons in the cultures. Similar effects can be observed after incubating with the conditioned medium derived from PPX- and ROP-treated substantia nigra astroglia. Meanwhile, PPX and ROP can protect the primary cells from insult of 1-methyl-4-phenylpyridinium (MPP+), the active metabolite of the neurotoxin 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPTP). Furthermore, the neurotrophic effects of PPX and ROP on mesencephalic dopamine neurons could be significantly blocked by D3 receptor antagonist, but not by D2 receptor antagonist. Moreover, we found that the levels of glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) in the conditioned medium of mesencephalic cultures treated with PPX and ROP were significantly increased. Blocking GDNF and BDNF with the neutralizing antibodies, the neurotrophic effects of PPX and ROP were greatly diminished. These results suggest that D3 dopamine receptor-preferring agonists, PPX and ROP, exert neurotrophic effects on cultured DA neurons by modulating the production of endogenous GDNF and BDNF, which may participate in their neuroprotection. PMID- 16307586 TI - Intrathecal minocycline attenuates peripheral inflammation-induced hyperalgesia by inhibiting p38 MAPK in spinal microglia. AB - Activation of p38 mitogen-activated protein kinase (p38) in spinal microglia is implicated in spinal nociceptive processing. Minocycline, a tetracycline derivative, displays selective inhibition of microglial activation, a function that is distinct from its antibiotic activity. In the present study we examined antinociceptive effects of intrathecal (IT) administration of minocycline in experimental models of inflammation-evoked hyperalgesia in addition to the effect of minocycline on stimulation-induced activation of p38 in spinal microglia. Intrathecal minocycline produced a dose-dependent reduction of formalin-evoked second-phase flinching behaviour in rats, and prevented thermal hyperalgesia induced by carrageenan injection into the paw. In contrast, systemic delivery (intraperitoneally) of minocycline inhibited the first but not the second phase of formalin-induced flinching, and it had no effect on carrageenan-induced hyperalgesia. Centrally mediated hyperalgesia induced by IT delivery of N-methyl d-aspartate was completely blocked by IT minocycline. An increase in phosphorylation (activation) of p38 (P-p38) was observed in the dorsal spinal cord after carrageenan paw injection, assessed by both Western blotting and immunohistochemistry. The increased P-p38 immunoreactivity was seen primarily in microglia but also in a small population of neurons. Minocycline, at the IT dose that blocked carrageenan-induced hyperalgesia, also attenuated the increased P p38 in microglia. In addition, minocycline suppressed lipopolysaccharide-evoked P p38 in cultured spinal microglial cells. Taken together, these findings show that minocycline given IT produces a potent and consistent antinociception in models of tissue injury and inflammation-evoked pain, and they provide strong support for the idea that this effect is mediated by direct inhibition of spinal microglia and subsequent activation of p38 in these cells. PMID- 16307587 TI - Post-conditioning with lipopolysaccharide reduces the inflammatory infiltrate to the injured brain and spinal cord: a potential neuroprotective treatment. AB - Systemic infection often accompanies or precedes acute brain injury, but it remains unclear how the systemic response contributes to outcome. To examine this problem we have microinjected recombinant interleukin-1beta (IL-1beta), a cytokine associated with acute brain injury, into the rat brain parenchyma and either preceded or followed this challenge with the intravenous injection of lipopolysaccharide (LPS), which mimics systemic inflammatory response syndrome. The microinjection of IL-1beta alone into the brain parenchyma gives rise to leukocyte mobilization in the blood, and to the delayed recruitment of neutrophils and monocytes to the brain with no evidence of blood-brain barrier breakdown or overt neuronal cell death. Systemic LPS pre-conditioning resulted in a dose-dependent reduction both in the number of circulating leukocytes and in the number of leukocytes recruited to the brain parenchyma after 12 h. Surprisingly, LPS given two hours after injury was equally effective in reducing the recruitment of leukocytes to the brain, which is more relevant to the management of clinical disease. In a more clinically relevant model of spinal cord injury, intravenous LPS post-conditioning also reduced the numbers of leukocytes mobilized in the blood and recruited to the spinal cord and thus limited the breakdown of the blood-spinal cord barrier. The effects appear to be specific to LPS, as they were not observed after intravenous IL-1beta pre conditioning. Our studies suggest that individual pro-inflammatory conditioning strategies may protect the injured central nervous system from the damaging consequences of leukocyte recruitment and may provide scope for novel therapeutic intervention. PMID- 16307588 TI - Imaging LTP of presynaptic release of FM1-43 from the rapidly recycling vesicle pool of Schaffer collateral-CA1 synapses in rat hippocampal slices. AB - Recent studies using the styryl dye FM1-43 and two-photon microscopy to directly visualize transmitter release at CA3-CA1 excitatory synapses in the hippocampus have demonstrated that activity-dependent long-term potentiation (LTP) and long term depression are associated with alterations in vesicular release. It is not known whether particular vesicle pools preferentially express these alterations or what second messenger cascades are involved. To address these questions, we selectively loaded FM1-43 into the rapidly recycling pool (RRP) of vesicles by use of a brief hypertonic shock to release and load the RRP. We demonstrate here that the induction of LTP can lead to a selective long-lasting enhancement in presynaptic release from the RRP, while reserve pool kinetics remain unchanged. LTP of RRP release was N-methyl-d-aspartate receptor-dependent and also required production of the intercellular messenger NO and activation of receptor tyrosine kinase. Measurement of FM1-43 stimulus-evoked uptake rates following induction of LTP confirmed that LTP produces more rapid recycling of vesicles released by electrical stimulation, consistent with an enhanced release probability from the RRP. PMID- 16307589 TI - Chronic nicotine exposure during adolescence differentially influences calcium binding proteins in rat anterior cingulate cortex. AB - We have recently shown that chronic amphetamine exposure selectively up-regulates parvalbumin (PV) calcium-binding proteins in the anterior cingulate cortex (ACC). In this study, we evaluated the effects of chronic nicotine (NIC) exposure on PV, calbindin D28k (CB) and calretinin (CR) calcium-binding protein immunostaining in ACC GABAergic interneurons. Chronic NIC exposure for 3 weeks in adolescent rats, either via drinking water (the oral group) or by twice daily subcutaneous injections (the injection group), resulted in the expression of high levels of CR proteins in the ACC but not in the parietal cortex. Larger increases in the density of CR-immunoreactive (ir) neurons were noted in the NIC-injected rats at 0-day withdrawal (45% increase) compared with the oral group (26% increase). The larger increases in CR-ir neuron density in the NIC-injected rats were also reflected by prominent CR-ir processes across cortical layers. The density of PV ir neurons was also increased (37%) at 0-day withdrawal but only in the oral NIC group and no changes in CB-ir neuron density were observed in either NIC group. Combined dual-immunofluorescence and confocal microscopy revealed that somatodendritic alpha4 nicotinic acetylcholine receptors colocalized with cortical neurons stained positively for CR, PV or CB. These results suggest that CR- and/or PV-ir-containing GABA interneurons may be involved in channeling the effects of NIC in the ACC, which is closely associated with the ventral basal ganglia circuit that is linked to brain reward function. PMID- 16307590 TI - Retinal abnormalities in human albinism translate into a reduction of grey matter in the occipital cortex. AB - Albinism is a genetic condition associated with abnormalities of the visual system. Defects in melanin production cause underdevelopment of the fovea, reduced retinal cell numbers and abnormal routing of ganglion cell nerve fibres at the optic chiasm. We examined 19 subjects with albinism and 26 control subjects to determine whether retinal abnormalities affect the structure of the visual cortex. Whole-brain, high-resolution anatomical magnetic resonance imaging volumes from each subject were obtained on a 1.5-T scanner and segmented into grey and white matter. A voxel-wise statistical comparison of grey and white matter volumes in the occipital lobes between the two groups was performed using voxel-based morphometry. Our analysis revealed a regionally specific decrease in grey matter volume at the occipital poles in albinism. The location of the decrease in grey matter corresponds to the cortical representation of the central visual field. This reduction is likely to be a direct result of decreased ganglion cell numbers in central retina in albinism. PMID- 16307591 TI - Protein kinase C immunoreactivity in the pigmented and albino rat retina. AB - The albino retina is abnormal. The central region is under-developed and some cell populations are reduced or increased in number. Not least of these anomalies is the deficit in the rod population in hypopigmented rodents and carnivores. Given this abnormality we have examined the distribution of rod bipolar cells in albino rats to determine whether this subsequent stage in the rod pathway is similarly disrupted. A monoclonal antibody to protein kinase C was used to determine the distribution of rod bipolar cells in juvenile and adult pigmented and albino rats. Immunoreactive rod bipolar cells and their processes were counted in transverse sections passing through both the central and peripheral retina. The mean densities of immunoreactive cells were significantly reduced in albino retinas at both juvenile (postnatal day 15) and adult stages, in the former by 14% and the latter by 9%. This was evident across the entire central-to peripheral extent of the retina. The reduced rod photoreceptor population found in albinos appears therefore to be consequential for the magnitude of their major target population, rod bipolar cells. The decrease in the rod bipolar population indicates a change in retinal cytoarchitecture and implies a disruption of functional organization of the albino retina, especially that underlying the scotopic channel. This, coupled with observations that some other retinal interneuronal populations may be disrupted, implies disordered retinal processing in albinos and emphasizes the likelihood that abnormal visual function in albinos may be as much a result of anomalous retinal circuitry as of the known photoreceptor deficit or chiasmatic misrouting. PMID- 16307592 TI - Reconfiguration of multiple motor networks by short- and long-term actions of an identified modulatory neuron. AB - The pyloric and gastric motor pattern-generating networks in the stomatogastric ganglion of the lobster Homarus gammarus are reconfigured into a new functional circuit by burst discharge in an identified pair of modulatory projection interneurons, originally named the pyloric suppressor (PS) neurons because of their inhibitory effects on pyloric network activity. Here we elucidate the actions of the PS neurons on individual members of the neighbouring gastric circuit, as well as describing their ability to alter synaptic coupling between the two networks. PS neuron firing has two distinct effects on gastric network activity: an initial short-lasting action mediated by transient inhibition of most gastric motoneurons, followed by a long-lasting circuit activation associated with a prolonged PS-evoked depolarization of the medial gastric (MG) motoneuron and the single network interneuron, Int1. These long-lasting effects are voltage-dependent, and experiments with hyperpolarizing current injection and photoablation suggest that excitation of both the MG neuron and Int1 is critical for PS-elicited gastric network rhythmicity. In parallel, PS neuron discharge persistently (lasting several minutes) enhances the strength of an inhibitory synaptic influence of the MG neuron on the pyloric dilator (PD)-anterior burster (AB) pacemaker neurons, thereby facilitating operational fusion of the two networks. Therefore, a single modulatory neuron may influence disparate populations of neurons via a range of very different and highly target-specific mechanisms: conventional transient synaptic drive and up- or down-modulation of membrane properties and synaptic efficacy. Moreover, distinctly different time courses of these actions allow different circuit configurations to be specified sequentially by a given modulatory input. PMID- 16307593 TI - Reciprocal connections between olfactory structures and the cortex of the rostral superior temporal sulcus in the Macaca fascicularis monkey. AB - Convergence of sensory modalities in the nonhuman primate cerebral cortex is still poorly understood. We present an anatomical tracing study in which polysensory association cortex located at the fundus and upper bank of the rostral superior temporal sulcus presents reciprocal connections with primary olfactory structures. At the same time, projections from this polysensory area reach multiple primary olfactory centres. Retrograde (Fast Blue) and anterograde (biotinylated dextran-amine and 3H-amino acids) tracers were injected into primary olfactory structures and rostral superior temporal sulcus. Retrograde tracers restricted to the anterior olfactory nucleus resulted in labelled neurons in the rostral portion of the upper bank and fundus of superior temporal sulcus. Injections of biotinylated dextran-amine at the fundus and upper bank of the superior temporal sulcus confirmed this projection by labelling axons in the dorsal and lateral portions of the anterior olfactory nucleus, as well as piriform, periamygdaloid and entorhinal cortices. Retrograde tracer injections at the rostral superior temporal sulcus resulted in neuronal labelling in the anterior olfactory nucleus, piriform, periamygdaloid and entorhinal cortices, thus providing confirmation of the reciprocity between primary olfactory structures and the cortex at the rostral superior temporal sulcus. The reciprocal connections between the rostral part of superior temporal sulcus and primary olfactory structures represent a convergence for olfactory and other sensory modalities at the cortex of the rostral temporal lobe. PMID- 16307594 TI - Projections from the hippocampal region to the mammillary bodies in macaque monkeys. AB - A combination of anterograde and retrograde tracers mapped the direct hippocampal and parahippocampal inputs to the mammillary bodies in two species of macaque monkey. Dense projections arose from pyramidal cells in layer III of the subiculum and prosubiculum, and terminated in the medial mammillary nucleus. While there was no evidence of an input from the dentate gyrus or fields CA1-3, a small contribution arose from the presubiculum and entorhinal cortices. All of the hippocampal and parahippocampal projections to the mammillary bodies appeared to use the fornix as a route. The caudal portions of the subiculum and prosubiculum contained the greatest numbers of cells projecting to the mammillary bodies. A light contralateral projection to the medial mammillary nucleus was also observed, although this appeared to arise primarily from the more rostral portions of the subiculum and prosubiculum. There was a crude topography within the medial mammillary nucleus, with the caudal subicular projections terminating in the mid and dorsal portions of the nucleus while the rostral subicular and entorhinal projections terminated in the ventral and lateral portions of the medial nucleus. Light ipsilateral projections throughout the lateral mammillary nucleus were sometimes observed. Comparisons with related studies of the macaque brain showed that the dense hippocampal projections to the mammillary bodies arise from a population of subicular cells separate from those that project to the anterior thalamic nuclei, even though the major output from the mammillary bodies is to the anterior thalamic nuclei. Other comparisons revealed underlying similarities with the corresponding projections in the rat brain. PMID- 16307595 TI - The suprachiasmatic nucleus controls the daily variation of plasma glucose via the autonomic output to the liver: are the clock genes involved? AB - In order to drive tissue-specific rhythmic outputs, the master clock, located in the suprachiasmatic nucleus (SCN), is thought to reset peripheral oscillators via either chemical and hormonal cues or neural connections. Recently, the daily rhythm of plasma glucose (characterized by a peak before the onset of the activity period) has been shown to be directly driven by the SCN, independently of the SCN control of rhythmic feeding behaviour. Indeed, the daily variation in glucose was not impaired unless the scheduled feeding regimen (six-meal schedule) was associated with an SCN lesion. Here we show that the rhythmicity of both clock-gene mRNA expression in the liver and plasma glucose is not abolished under such a regular feeding schedule. Because the onset of the activity period and hyperglycemia are correlated with an increased sympathetic tonus, we investigated whether this autonomic branch is involved in the SCN control of plasma glucose rhythm and liver rhythmicity. Interestingly, hepatic sympathectomy combined with a six-meal feeding schedule resulted in a disruption of the plasma glucose rhythmicity without affecting the daily variation in clock-gene mRNA expression in the liver. Taking all these data together, we conclude that (i) the SCN needs the sympathetic pathway to the liver to generate the 24-h rhythm in plasma glucose concentrations, (ii) rhythmic clock-gene expression in the liver is not dependent on the sympathetic liver innervation and (iii) clock-gene rhythmicity in liver cells is not sufficient for sustaining a circadian rhythm in plasma glucose concentrations. PMID- 16307596 TI - GABAergic synaptic transmission modulates swimming in the ascidian larva. AB - To examine the role of the amino acid GABA in the locomotion of basal chordates, we investigated the pharmacology of swimming and the morphology of GABA immunopositive neurones in tadpole larvae of the ascidians Ciona intestinalis and Ciona savignyi. We verified that electrical recording from the tail reflects alternating muscle activity during swimming by correlating electrical signals with tail beats using high-speed video recording. GABA reversibly reduced swimming periods to single tail twitches, while picrotoxin increased the frequency and duration of electrical activity associated with spontaneous swimming periods. Immunocytochemistry for GABA revealed extensive labelling throughout the larval central nervous system. Two strongly labelled regions on either side of the sensory vesicle were connected by an arc of labelled fibres, from which fibre tracts extended caudally into the visceral ganglion. Fibre tracts extended ventrally from a third, more medial region in the posterior sensory vesicle. Two rows of immunoreactive cell bodies in the visceral ganglion extended neurites into the nerve cord, where varicosities were seen. Thus, presumed GABAergic neurones form a network that could release GABA during swimming that is involved in modulating the time course and frequency of periods of spontaneous swimming. GABAergic and motor neurones in the visceral ganglion could interact at the level of their cell bodies and/or through the presumed GABAergic fibres that enter the nerve cord. The larval swimming network appears to possess some of the properties of spinal networks in vertebrates, while at the same time possibly showing a type of peripheral innervation resembling that in some protostomes. PMID- 16307597 TI - A lateralized avian hippocampus: preferential role of the left hippocampal formation in homing pigeon sun compass-based spatial learning. AB - The hippocampal formation (HF) plays a crucial role in amniote spatial cognition. There are also indications of functional lateralization in the contribution of the left and right HF in processes that enable birds to navigate space. The experiments described in this study were designed to examine left and right HF differences in a task of sun compass-based spatial learning in homing pigeons (Columba livia). Control, left (HFL) and right (HFR) HF lesioned pigeons were trained in an outdoor arena to locate a food reward using their sun compass in the presence or absence of alternative feature cues. Subsequent to training, the pigeons were subjected to test sessions to determine if they learned to represent the goal location with their sun compass and the relative importance of the sun compass vs. feature cues. Under all test conditions, the control pigeons demonstrated preferential use of the sun compass in locating the goal. By contrast, the HFL pigeons demonstrated no ability to locate the goal by the sun compass but an ability to use the feature cues. The behaviour of the HFR pigeons demonstrated that an intact left HF is sufficient to support sun compass-based learning, but in conflict situations and in contrast to controls, they often relied on feature cues. In conclusion, only the left HF is capable of supporting sun compass-based learning. However, preferential use of the sun compass for learning requires an intact right HF. The data support the hypothesis that the left and right HF make different but complementary contributions toward avian spatial cognition. PMID- 16307598 TI - A molecular switch for translational control in taste memory consolidation. AB - In a variety of species memory consolidation following different learning paradigms has been shown to be dependent on protein synthesis. However, it is not known whether modulation of protein synthesis is a critical component of the consolidation process, nor is the identity of any protein(s) subject to translational regulation, known. We report here that phosphorylation of eukaryotic elongation factor-2 (eEF2), an indicator for translational elongation attenuation, is correlated with input that produces taste memory consolidation in the relevant cortex of rat. The temporal pattern of eEF2 phosphorylation is similar to extra-cellular regulated kinase 2 (ERK2) activation and S6K1 phosphorylation, which are known to stimulate translation initiation. In addition, increased eEF2 phosphorylation and increased alphaCaMKII expression is detected in a synaptoneurosomal fraction made from taste cortex following memory consolidation. These results suggest that increased initiation rate together with decreased elongation rate, during memory consolidation, shift the rate-limiting step of protein synthesis, to produce a local switch-like effect in the expression of neuronal proteins. PMID- 16307599 TI - Tissue inhibitor of metalloproteinases-1 (TIMP-1) modulates neuronal death, axonal plasticity, and learning and memory. AB - The tissue inhibitor of metalloproteinases-1 (TIMP-1) belongs to a family of multifunctional proteins that inhibit matrix metalloproteinases (MMPs), but also regulate cell growth, proliferation, migration and apoptosis in non-nervous tissues. We had previously reported that kainate (KA)-mediated excitotoxic seizures induce the expression of TIMP-1 in resistant neurons and reactive astrocytes of the rat CNS, but the functional implications of these changes had not been elucidated. In the present work we used a targeted gene null mutation in mice to investigate in vivo the involvement of TIMP-1 in neuronal death and axonal sprouting following KA. We found no differences in seizure behaviour between the wild-type (WT) and the TIMP-1 knock-out (KO) mice, without any compensation by other TIMPs, at least at the mRNA level. However, the TIMP-1 KO mice were resistant to excitotoxicity and did not undergo the typical mossy fibre sprouting observed in WT mice. The lack of TIMP-1 paradoxically hampered the increase in the activity of MMPs observed in the seizing WT mice. In addition, we demonstrate that learning and memory are impaired in untreated KO mice. In conclusion, this study provides the first in vivo evidence for the implication of TIMP-1 in neuronal death and axonal sprouting in a pathological situation, but also suggests the involvement of TIMP-1 in the synaptic mechanisms underlying learning and memory in physiological conditions. More generally, these data support the idea that the control of proteolysis is instrumental for pathological and physiological processes in the brain. PMID- 16307600 TI - Consequences of partial and severe dopaminergic lesion on basal ganglia oscillatory activity and akinesia. AB - Severe chronic dopamine (DA) depletion increases the proportion of neurons in the basal ganglia that fire rhythmic bursts of action potential (LFO units) synchronously with the cortical oscillations. Here we report on how different levels of mesencephalic DA denervation affect substantia nigra pars reticulata (SNpr) neuronal activity in the rat and its relationship to akinesia (stepping test). Chronic nigrostriatal lesion induced with 0 (control group), 4, 6 or 8 microg of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle resulted in a dose-dependent decrease of tyrosine hydroxylase positive (TH+) neurons in the SN and ventral tegmental area (VTA). Although 4 microg of 6-OHDA reduced the number of TH+ neurons in the SN by approximately 60%, both stepping test performance and SNpr neuronal activity remained indistinguishable from control animals. By contrast, animals that received 6 microg of 6-OHDA showed a marked reduction of TH+ cells in the SN ( approximately 75%) and VTA ( approximately 55%), a significant stepping test deficit and an increased proportion of LFO units. These changes were not dramatically enhanced with 8 microg 6-OHDA, a dose that induced an extensive DA lesion (> 95%) in the SN and approximately 70% reduction of DA neurons in the VTA. These results suggest a threshold level of DA denervation for both the appearance of motor deficits and LFO units. Thus, the presence of LFO activity in the SNpr is not related to a complete nigrostriatal DA neuron depletion (ultimate stage parkinsonism); instead, it may reflect a functional disruption of cortico-basal ganglia dynamics associated with clinically relevant stages of the disease. PMID- 16307601 TI - Lentivector-mediated delivery of GDNF protects complex motor functions relevant to human Parkinsonism in a rat lesion model. AB - Although viral vector-mediated delivery of glial cell-line derived neurotrophic factor (GDNF) to the brain has considerable potential as a neuroprotective strategy in Parkinson's disease (PD), its ability to protect complex motor functions relevant to the human condition has yet to be established. In this study, we used an operant task that assesses the selection, initiation and execution of lateralized nose-pokes in Lister Hooded rats to assess the efficacy with which complex behaviours are protected against neurotoxic lesions by prior injection of a lentiviral vector expressing GDNF. Unilateral injection of 6 hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) caused rats to attempt fewer trials and to make more procedural errors. Lesioned rats also developed a pronounced ipsilateral bias, with a corresponding drop in contralateral accuracy. They were also slower to react to contralateral stimuli and to execute movements bilaterally. Rats that were pre-treated 4 weeks prior to lesion surgery with an equine infectious anaemia virus (EIAV) vector carrying GDNF [EIAV-GDNF, injected into the striatum and above the substantia nigra (SN)] performed significantly better on all of these parameters than control rats. In addition to the operant task, EIAV-GDNF successfully rescued contralateral impairments in the corridor, staircase, stepping and cylinder tasks, and prevented drug-induced rotational asymmetry. This study confirms that GDNF can protect against 6-OHDA-induced impairments in complex as well as simple behaviours, and reinforces the use of EIAV-based vectors for the treatment of PD. PMID- 16307602 TI - Basolateral amygdala glutamatergic activation enhances taste aversion through NMDA receptor activation in the insular cortex. AB - In conditioned taste aversion (CTA), a subject learns to associate a novel taste with visceral malaise. Brainstem, limbic and neocortical structures have been implicated in CTA memory formation. Nevertheless, the role of interactions between forebrain structures during these processes is still unknown. The present experiment was aimed at investigating the possible interaction between the basolateral nucleus of the amygdala (BLA) and the insular cortex (IC) during CTA memory formation. Injection of a low dose of lithium chloride (30 mg/kg, i.p.) 30 min after novel taste consumption (saccharin 0.1%) induces a weak CTA. Unilateral BLA injection of glutamate (2 microg in 0.5 microL) just before low lithium induces a stronger CTA. Unilateral injection of an N-methyl-d-aspartate (NMDA) receptor antagonist (AP5, 5 microg in 0.5 microL) in IC has no effect. However, AP5 treatment in IC at the same time or 1 h after the ipsilateral BLA injection reverses the glutamate-induced CTA enhancement. Injection of AP5 in IC 3 h after BLA injection does not interfere with the glutamate effect. Moreover, the CTA enhancing effect of glutamate was also blocked by contralateral IC injection of AP5 at the same time. These results provide strong evidence that NMDA receptor activation in the IC is essential to enable CTA enhancement induced by glutamate infusion in the BLA during a limited time period that extends to 1 but not to 3 hours. These findings indicate that BLA-IC interactions regulate the strength of CTA. The bilateral nature of these amygdalo-cortical interactions is discussed. PMID- 16307603 TI - Double dissociation of the effects of selective nucleus accumbens core and shell lesions on impulsive-choice behaviour and salience learning in rats. AB - The nucleus accumbens can be subdivided into at least two anatomically distinct subregions: a dorsolateral 'core' and a ventromedial 'shell', and this distinction may extend to a functional dissociation. Here, we contrasted the effects of selective excitotoxic core and medial shell lesions on impulsive choice behaviour using a delayed reward choice paradigm and a differential reward for low rates of responding (DRL) test, against a form of salience learning known as latent inhibition (LI). Core lesions led to enhanced impulsive choices as evidenced by a more pronounced shift from choosing a continuously reinforced lever to a partially reinforced lever, when a delay between lever press and reward delivery was imposed selectively on the former. The core lesions also impaired performance on a DRL task that required withholding the response for a fixed period of time in order to earn a reward. Medial shell lesions had no effect on these two tasks, but abolished the LI effect, as revealed by the failure of stimulus pre-exposure to retard subsequent conditioning to that stimulus in an active avoidance procedure in the lesioned animals. As expected, selective core lesions spared LI. The double dissociations demonstrated here support a functional segregation between nucleus accumbens core and shell, and add weight to the hypothesis that the core, but not the shell, subregion of the nucleus accumbens is preferentially involved in the control of choice behaviour under delayed reinforcement conditions and in the inhibitory control of goal directed behaviour. PMID- 16307604 TI - Ventral pallidal neurons code incentive motivation: amplification by mesolimbic sensitization and amphetamine. AB - Neurons in ventral pallidum fire to reward and its predictive cues. We tested mesolimbic activation effects on neural reward coding. Rats learned that a Pavlovian conditioned stimulus (CS+1 tone) predicted a second conditioned stimulus (CS+2 feeder click) followed by an unconditioned stimulus (UCS sucrose reward). Some rats were sensitized to amphetamine after training. Electrophysiological activity of ventral pallidal neurons to stimuli was later recorded under the influence of vehicle or acute amphetamine injection. Both sensitization and acute amphetamine increased ventral pallidum firing at CS+2 (population code and rate code). There were no changes at CS+1 and minimal changes to UCS. With a new 'Profile Analysis', we show that mesolimbic activation by sensitization/amphetamine incrementally shifted neuronal firing profiles away from prediction signal coding (maximal at CS+1) and toward incentive coding (maximal at CS+2), without changing hedonic impact coding (maximal at UCS). This pattern suggests mesolimbic activation specifically amplifies a motivational transform of CS+ predictive information into incentive salience coded by ventral pallidal neurons. Our results support incentive-sensitization predictions and suggest why cues temporally proximal to drug presentation may precipitate cue triggered relapse in human addicts. PMID- 16307605 TI - Delaying visually guided saccades by microstimulation of macaque V1: spatial properties of delay fields. AB - Electrical microstimulation of macaque primary visual cortex (area V1) is known to delay the execution of saccadic eye movements made to a punctate visual target placed into the receptive field of the stimulated neurons. We examined the spatial extent of this delay effect, which we call a delay field, by placing a 0.2 degrees visual target at various locations relative to the receptive field of the stimulated neurons and by stimulating different sites within the operculum of V1. A 100-ms train of stimulation consisting of current pulses at or less than 100 microA was delivered immediately before monkeys generated a saccadic eye movement to the visual target. The region of tissue activated was within 0.5 mm from the electrode tip. The depth of stimulation for a given site ranged from 0.9 to 2.0 mm below the cortical surface. The location of the receptive fields of the stimulated neurons ranged from 1.8 to 4.4 degrees of eccentricity from the center of gaze. Within this range, the size of the delay field increased from 0.1 to 0.55 degrees of visual angle. The shape of the field was roughly circular. The size of the delay field increased as the stimulation site was located further from the foveal representation of V1. These results are consistent with the finding that phosphenes evoked by electrical stimulation of human V1 are circular and increase in size as the stimulating electrode is placed more distant from the foveal representation of V1. PMID- 16307606 TI - GABARAP is not essential for GABA receptor targeting to the synapse. AB - GABA(A) receptors (GABA(A)Rs) containing the gamma2 subunit are thought to require the interacting protein GABARAP (GABA(A)R associated protein) for trafficking to the neuronal plasma membrane. In order to assess whether GABARAP is required for GABA(A) receptor accumulation at synaptic sites, we analysed a GABARAP knockout mouse. GABARAP deficient mice are phenotypically normal and do not show up-regulation of other GABARAP homologues. Also, the total number of GABA(A)Rs, as assessed by benzodiazepine binding, is unaffected by the loss of GABARAP. Immunocytochemistry of cortical sections showed no differences in the expression and punctate distribution of the gamma2 subunit and the receptor anchoring protein gephyrin between GABARAP deficient and wild-type mice. Thus, GABARAP is not essential for trafficking gamma2 subunit containing GABA(A)Rs to the neuronal plasma membrane or targeting them to inhibitory synapses. PMID- 16307607 TI - The Grueneberg ganglion of the mouse projects axons to glomeruli in the olfactory bulb. AB - First described in 1973, the Grueneberg ganglion (GG) is an arrow-shaped neuronal structure at the anterior end of the nasal cavity. It lines both sides of the nasal septum, within the nasal vestibule, close to the opening of the naris. The functions of the GG and the pattern of projections to the brain are not known. Here, we report that neurons of the mouse GG express olfactory marker protein, which is normally expressed in mature olfactory or vomeronasal sensory neurons. The approx. 500 cells in each GG are arranged in several densely packed cell clusters. Individual cells give rise to single axons, which fasciculate to form a nerve bundle that projects caudally. The axons terminate in glomeruli of the olfactory bulb, one or two large glomeruli associated with a semicircle of up to 10 smaller, somewhat diffusely organized glomeruli that surround the most anterior part of the accessory olfactory bulb. Development of the GG starts around embryonic day 16 and appears to be completed at birth; cell numbers then undergo a minor decrease during postnatal development. The strategic location of the GG, expression of olfactory marker protein, axonal projections to glomeruli at particular locations in the olfactory bulb and early development suggest that this neuronal structure performs specific chemosensory functions at neonatal stages. PMID- 16307608 TI - Modulation of calcium currents in mouse ventral horn neurons by extracellular pH. AB - Neuronal activity has been shown to modulate the pH of the extracellular environment. Since neuronal circuits in the ventral horn of the spinal cord are highly active during patterned movements, and voltage-gated calcium channels play an important role in the production of spinal motoneuron output, the effects of changes in extracellular pH (pH(e)) on calcium currents in ventral horn neurons of the mouse spinal cord were examined. It is demonstrated that these channels are sensitive to modulation by pH(e). The amplitude of the current mediated by these channels increased as the pH(e) was elevated. The elevated pH(e) also led to a hyperpolarizing shift in the voltage dependence of both activation and inactivation. The opposite effects were seen for a decrease in pH(e). It was also noted that a decrease in pH(e) was associated with a faster inactivation of the current. It is concluded that voltage-gated calcium currents in ventral horn neurons are modulated by changes in pH(e), and that this modulation may play a physiologically important role in determining motoneuronal excitability during behaviors such as locomotion. PMID- 16307609 TI - Electrical coupling induces bistability of rhythms in networks of inhibitory spiking neurons. AB - Information processing in higher brain structures is thought to rely on the synchronization of spiking neurons. Increasing evidence indicates that, within these structures, inhibitory neurons are linked by both chemical and electrical synapses. However, how synchronized states may emerge from such circuits is not fully understood. Using snail neurons interconnected through a dynamic-clamp system, we show that networks of spiking neurons linked by both reciprocal inhibition and electrical coupling can express two coexisting coordination patterns of different rhythms. One of these patterns consists of antiphase firing of the network partners whereas, in the other, neurons fire synchronously. Switching between patterns may be evoked immediately by transient stimuli, demonstrating bistability of the network. Thus electrical coupling can provide a potent way for instantaneous reconfiguration of activity patterns in inhibitory spiking networks without alteration of intrinsic network properties by modulatory processes. PMID- 16307610 TI - Synchronized network activity in developing rat hippocampus involves regional hyperpolarization-activated cyclic nucleotide-gated (HCN) channel function. AB - The principal form of synchronized network activity in neonatal hippocampus consists of low frequency 'giant depolarizing potentials' (GDPs). Whereas contribution of both GABA and glutamate to their generation has been demonstrated, full understanding of the mechanisms underlying these synchronized activity bursts remains incomplete. A contribution of the h-current, conducted by HCN channels, to GDPs has been a topic of substantial interest. Here we focus on HCN1, the prevalent HCN channel isoform in neonatal hippocampus, and demonstrate an HCN1 spatiotemporal expression pattern in both CA3 principal cells and interneurons that correlates with the developmental profile of GDPs. Abrogation of HCN physiological function in CA3, via the selective I(h)-blocker ZD7288, disrupts GDP generation. Furthermore, ZD7288 specifically abolishes spontaneous bursting of the CA3 pyramidal cells at frequencies typical of GDPs without major influence on interneuronal firing. These findings support a pivotal role for HCN channels expressed by CA3 neurons, and particularly CA3 pyramidal cells, in GDP related network synchronization. PMID- 16307611 TI - m-Chlorophenylpiperazine exacerbates perception of salicylate-induced tinnitus in rats. AB - The serotonin receptor 5-HT2c agonist m-chlorophenylpiperazine (mCPP) has been widely used to induce anxiety-like states in animals and anxiety in humans. We here addressed the effect of an intraperitoneal injection of mCPP on the perception of tinnitus in a behavioural protocol based on an active avoidance paradigm. In control saline-treated animals, mCPP did not change compound action potential audiograms or measurable tinnitus. In contrast, mCPP to animals demonstrating salicylate-induced tinnitus exacerbated tinnitus perception by nearly twofold. We went on to test whether manipulation of the peripheral generator of tinnitus (i.e. the cochlea) could extinguish this exacerbated perception by applying the N-methyl-d-aspartate (NMDA) antagonist 7 chlorokynurenate (7-CK) into cochlear fluids using a 7-CK-soaked Gelfoam ball placed on the round window of each of the animals' two ears. In addition to blocking the tinnitus induced by salicylate alone, 50 microm of 7-CK clearly abolished tinnitus in animals receiving salicylate and an injection of mCPP. The demonstration that cochlear NMDA receptor blockade abolishes the exacerbated perception of tinnitus is highly relevant in terms of treatment. In addition to psychotherapeutic treatment that may help to attenuate an individual's perception of tinnitus, targeting cochlear NMDA receptors represents a promising therapeutic strategy, even in depressed or chronically anxious patients. PMID- 16307614 TI - Antidepressant combinations: epidemiological considerations. AB - OBJECTIVE: To evaluate the treatment options in patients who do not respond appropriately to a single antidepressant alone. METHOD: The medical literature was reviewed. RESULTS: A number of strategies are available if a patient fails to respond adequately to initial antidepressant treatment, including the combination with another psychoactive drug. Evidence published to date appears to suggest that benzodiazepines are the drugs most frequently combined with antidepressants. The combination of two antidepressants together is less common, occurring in approximately 5-15% of cases showing a poor initial response. The key figures involved in such co-prescription are psychiatrists. CONCLUSION: There appears to be considerable variability in the data concerning combined prescription of antidepressants, with differences arising depending on the type of physician, the type of patient or illness and the geographical area. It is also unclear how closely research findings parallel with what doctors do in everyday practice. PMID- 16307615 TI - Neuropharmacological basis of combining antidepressants. AB - OBJECTIVE: To review the neuropharmacological basis of antidepressant combination therapy. METHOD: Literature searches and other relevant material were obtained and reviewed. RESULTS: The overall clinical aim of combining antidepressants is to increase the efficacy whilst minimizing the side effects. Although such prescriptions are frequently based on the previous experience and knowledge, a sound neuropharmacological basis to support these combinations is desirable. When combining antidepressants, it is important to combine mechanisms of action, rather than simply one drug with another, and to aim for synergistic effects. The possibilities of combining mechanisms of action should also be exploited to the full if necessary, and the potential exists for combining two independent actions that have synergistic effects on the serotonergic, noradrenergic and even the dopaminergic systems. CONCLUSION: Unfortunately, there are still, as yet, insufficient data to categorically justify choosing one or other combination based only on the neuropharmacological evidence. PMID- 16307616 TI - Potentiation strategies for treatment-resistant depression. AB - OBJECTIVE: To review the pharmacological basis of antidepressant potentiation in combination therapy and the clinical evidence for its efficacy. METHOD: Literature searches were undertaken and the results reviewed. RESULTS: Treatment resistant depression is common (15-30%). Various strategies exist for dealing with resistant depression, including pharmacological potentiation, i.e. adding a treatment that itself does not have antidepressant actions but that enhances the efficacy of the original treatment. Lithium, triiodothyronine (T3) and buspirone are the best studied potentiating drugs, although other options include pindolol, dopaminergic agents, second-generation antipsychotics, psychostimulants, hormones and anticonvulsants. CONCLUSION: Several pharmacological potentiation strategies exist. Whilst good evidence exists for lithium combined with antidepressants, although good results have also been reported with augmentation strategies involving T3 or buspirone. PMID- 16307617 TI - Combined antidepressants: clinical experience. AB - OBJECTIVE: To review the current literature on the use of combinations of antidepressive agents. METHOD: Literature searches were undertaken and reviewed on the use of combinations of antidepressants. RESULTS: Data sources included surveys, analyses of prescription records, decision algorithms, clinical reports, and studies comparing the monotherapy with combination therapy. More recent surveys recommend combining different selective serotonin reuptake inhibitors (SSRIs), an SSRI plus bupropion or dual action antidepressants plus an SSRI. Decision algorithms recommend an SSRI plus tricyclic antidepressant (TCA) and more recently bupropion plus venlafaxine or mirtazapine. Few controlled clinical trials comparing the combined therapy with monotherapy have been conducted. Beneficial effects have been reported with combinations of TCAs plus mianserin or SSRIs plus mirtazapine. CONCLUSION: Adding or combining antidepressant medications has advantages for the speed of onset and maintaining the existing response. More rigorous clinical trials comparing combination therapy with monotherapy and for the development of rational treatment guidelines are required. PMID- 16307618 TI - Use of antidepressant combinations: which, when and why? Results of a Spanish survey. AB - OBJECTIVE: The present study uses the data from a large survey conducted to examine the general practice of Spanish psychiatrists on the use of antidepressant combinations in the treatment of depressive disorders. METHOD: The sample was drawn from specialists and psychiatric residents practicing in Spain who were respondents to a questionnaire distributed during an annual national psychiatry meeting and sent by mail. RESULTS: A total of 1032 questionnaires were collected; following the data-filtering, 831 were analysed. Most psychiatrists (89%) believe that many patients do not respond to the first treatment; in such cases of non-response, 58% choose a combination of antidepressants as the next treatment option. Reasons for using the combined treatments include greater efficacy (57%), overcoming resistance to the first antidepressant (27%), faster onset of action (21%) and avoidance of side effects (17%). The most sought after pharmacological profile was serotonergic-noradrenergic (96%) and the most popular combinations were selective serotonin reuptake inhibitor (SSRI) + mirtazapine, SSRI + reboxetine and SSRI + tricyclic antidepressant. CONCLUSION: Antidepressant combinations are frequently used in clinical practice. Pharmacological profiles are always considered and SSRIs + mirtazapine is the option usually chosen. PMID- 16307620 TI - Knowledge and perceptions of colorectal cancer screening among urban African Americans. AB - OBJECTIVE: To explore colorectal cancer (CRC) screening knowledge, attitudes, barriers, and preferences among urban African Americans as a prelude to the development of culturally appropriate interventions to improve screening for this group. DESIGN: Qualitative focus group study with assessment of CRC screening preferences. SETTING: Community health center serving low-income African Americans. PARTICIPANTS: Fifty-five self-identified African Americans over 40 years of age. MEASUREMENTS AND MAIN RESULTS: Transcripts were analyzed using an iterative coding process with consensus and triangulation on final thematic findings. Six major themes were identified: (1) Hope--a positive attitude toward screening, (2) Mistrust--distrust that the system or providers put patients first, (3) Fear--fear of cancer, the system, and of CRC screening procedures, (4) Fatalism--the belief that screening and treatment may be futile and surgery causes spread of cancer, (5) Accuracy--a preference for the most thorough and accurate test for CRC, and (6) Knowledge--lack of CRC knowledge and a desire for more information. The Fear and Knowledge themes were most frequently noted in transcript theme counts. The Hope and Accuracy themes were crucial moderators of the influence of all barriers. The largest number of participants preferred either colonoscopy (33%) or home fecal occult blood testing (26%). CONCLUSIONS: Low-income African Americans are optimistic and hopeful about early CRC detection and believe that thorough and accurate CRC screening is valuable. Lack of CRC knowledge and fear are major barriers to screening for this population along with mistrust, and fatalism. PMID- 16307621 TI - Using a computer to teach patients about fecal occult blood screening. A randomized trial. AB - OBJECTIVE: To determine whether a multimedia computer program could effectively teach patients about fecal occult blood testing (FOBT) and increase screening rates. DESIGN: Randomized trial. SETTING: University-affiliated, community-based Internal Medicine outpatient practice. PARTICIPANTS: All English-speaking patients aged 50 years and older who were offered FOBT screening by their providers were invited to participate. Two hundred and four patients enrolled in the study. Ten patients were later determined to be ineligible. INTERVENTIONS: Patients were randomized to either the educational multimedia computer program or usual nurse counseling about FOBT screening. Screening instructions were based on the material pre-printed on each test kit. Educational sessions were held in a private setting immediately after each patient's office visit. MEASUREMENTS AND MAIN RESULTS: A knowledge-assessment questionnaire was administered in a blinded fashion by telephone the following day. Successful screening was defined as return of the test kits within 30 d. Completion of the FOBT kits was similar in both groups: 62% (58/93) in the computer group and 63% (64/101) in the nurse group (P=.89). Mean knowledge scores were also similar, but there was a trend toward increased knowledge mastery in the computer group (56% vs 41%, P=.09). CONCLUSIONS: A multimedia educational computer program was as effective as usual nurse counseling in educating patients and achieving adherence to FOBT screening. Future studies are needed to determine whether computer-assisted instruction can improve health outcomes. PMID- 16307622 TI - Predictors of nonadherence to screening colonoscopy. AB - BACKGROUND: Colonoscopy has become a preferred colorectal cancer (CRC) screening modality. Little is known about why patients who are referred for colonoscopy do not complete the recommended procedures. Prior adherence studies have evaluated colonoscopy only in combination with flexible sigmoidoscopy, failed to differentiate between screening and diagnostic procedures, and have examined cancellations/no-shows, but not nonscheduling, as mechanisms of nonadherence. METHODS: Sociodemographic predictors of screening completion were assessed in a retrospective cohort of 647 patients referred for colonoscopy at a major university hospital. Then, using a qualitative study design, a convenience sample of patients who never completed screening after referral (n=52) was interviewed by telephone, and comparisons in reported reasons for nonadherence were made by gender. RESULTS: Half of all patients referred for colonoscopy failed to complete the procedure, overwhelmingly because of nonscheduling. In multivariable analysis, female sex, younger age, and insurance type predicted poorer adherence. Patient-reported barriers to screening completion included cognitive-emotional factors (e.g., lack of perceived risk for CRC, fear of pain, and concerns about modesty and the bowel preparation), logistic obstacles (e.g., cost, other health problems, and competing demands), and health system barriers (e.g., scheduling challenges, long waiting times). Women reported more concerns about modesty and other aspects of the procedure than men. Only 40% of patients were aware of alternative screening options. CONCLUSIONS: Adherence to screening colonoscopy referrals is sub-optimal and may be improved by better communication with patients, counseling to help resolve logistic barriers, and improvements in colonoscopy referral and scheduling mechanisms. PMID- 16307623 TI - Patients and medical statistics. Interest, confidence, and ability. AB - BACKGROUND: People are increasingly presented with medical statistics. There are no existing measures to assess their level of interest or confidence in using medical statistics. OBJECTIVE: To develop 2 new measures, the STAT-interest and STAT-confidence scales, and assess their reliability and validity. DESIGN: Survey with retest after approximately 2 weeks. SUBJECTS: Two hundred and twenty-four people were recruited from advertisements in local newspapers, an outpatient clinic waiting area, and a hospital open house. MEASURES: We developed and revised 5 items on interest in medical statistics and 3 on confidence understanding statistics. RESULTS: Study participants were mostly college graduates (52%); 25% had a high school education or less. The mean age was 53 (range 20 to 84) years. Most paid attention to medical statistics (6% paid no attention). The mean (SD) STAT-interest score was 68 (17) and ranged from 15 to 100. Confidence in using statistics was also high: the mean (SD) STAT-confidence score was 65 (19) and ranged from 11 to 100. STAT-interest and STAT-confidence scores were moderately correlated (r=.36, P<.001). Both scales demonstrated good test-retest repeatability (r=.60, .62, respectively), internal consistency reliability (Cronbach's alpha=0.70 and 0.78), and usability (individual item nonresponse ranged from 0% to 1.3%). Scale scores correlated only weakly with scores on a medical data interpretation test (r=.15 and .26, respectively). CONCLUSION: The STAT-interest and STAT-confidence scales are usable and reliable. Interest and confidence were only weakly related to the ability to actually use data. PMID- 16307624 TI - Physician notification of their diabetes patients' limited health literacy. A randomized, controlled trial. AB - BACKGROUND: Many patients with chronic disease have limited health literacy (HL). Because physicians have difficulty identifying these patients, some experts recommend instituting screening programs in clinical settings. It is unclear if notifying physicians of patients' limited HL improves care processes or outcomes. OBJECTIVE: To determine whether notifying physicians of their patients' limited HL affects physician behavior, physician satisfaction, or patient self-efficacy. DESIGN: We screened all patients for limited HL and randomized physicians to be notified if their patients had limited HL skills. PARTICIPANTS: Sixty-three primary care physicians affiliated with a public hospital and 182 diabetic patients with limited HL. MEASUREMENTS: After their visit, physicians reported their management strategies, satisfaction, perceived effectiveness, and attitudes toward HL screening. We also assessed patients' self-efficacy, feelings regarding HL screening's usefulness, and glycemic control. RESULTS: Intervention physicians were more likely than control physicians to use management strategies recommended for patients with limited HL (OR 3.2, P=.04). However, intervention physicians felt less satisfied with their visits (81% vs 93%, P=.01) and marginally less effective (38% vs 53%, P=.10). Intervention and control patients' post-visit self efficacy scores were similar (12.6 vs 12.9, P=.6). Sixty-four percent of intervention physicians and 96% of patients felt HL screening was useful. CONCLUSIONS: Physicians are responsive to receiving notification of their patients' limited HL, and patients support the potential utility of HL screening. However, instituting screening programs without specific training and/or system wide support for physicians and patients is unlikely to be a powerful tool in improving diabetes outcomes. PMID- 16307625 TI - Bleeding Risk Index in an anticoagulation clinic. Assessment by indication and implications for care. AB - BACKGROUND: The Outpatient Bleeding Risk Index (BRI) prospectively classified patients who were at high, intermediate, or low risk for warfarin-related major bleeding. However, there are only 2 published validation studies of the index and neither included veterans. OBJECTIVE: To determine the accuracy of the BRI in patients attending a Veterans Affairs (VA) anticoagulation clinic and to specifically evaluate the accuracy of the BRI in patients with atrial fibrillation. DESIGN: Retrospective cohort study. PATIENTS AND MEASUREMENTS: Using the BRI, all patients managed by the Anticoagulation Clinic between January 1, 2001 and December 31, 2002 were classified as high, intermediate, or low risk for major bleeding. Bleeds were identified via quality-assurance reports. Poisson regression was used to determine whether there was an association between the index and the development of bleeding. RESULTS: The rate of major bleeding was 10.6%, 2.5%, and 0.8% per patient-year of warfarin in the high-, intermediate-, and low-risk groups, respectively. Patients in the high-risk category had 14 times the rate of major bleeding of those in the low-risk group (incidence rate ratio (IRR) 14; 95% confidence interval (CI), 1.9 to 104.7). The rate of major bleeding was significantly different between the high- and intermediate-risk categories (P<.001). Among those with atrial fibrillation, patients in the high risk category had 6 times the major bleeding rate of those in the intermediate- and low-risk groups combined (IRR=6; 95% CI, 2.4 to 15.3). CONCLUSIONS: The BRI discriminates between high- and intermediate-risk patients in a VA anticoagulation clinic, including those with atrial fibrillation. PMID- 16307626 TI - Mentorship in academic general internal medicine. Results of a survey of mentors. AB - BACKGROUND: Effective mentorship is crucial to career development. Strategies to improve the availability of mentors include mentoring multiple mentees at once, compensating mentors, comentoring, and long-distance mentoring. OBJECTIVE: To describe current trends in mentorship in general Internal Medicine (GIM). METHODS: We conducted a national cross-sectional web-based survey of GIM mentors, GIM fellowship directors, and GIM National Institutes of Health K24 grant awardees to capture their experiences with mentoring, including compensation for mentorship, multiple mentees, comentorship, and long-distance mentorship. We compared experiences by mentorship funding status, faculty type, academic rank, and sex. RESULTS: We collected data from 111 mentors (77% male, 54% full professors, and 68% clinician-investigators). Fifty-two (47%) received funding for mentorship. Mentors supervised a median (25th percentile, 75th percentile) of 5 (3, 8) mentees each, and would be willing to supervise a maximum of 6 (4, 10) mentees at once. Compared with mentors without funding, mentors with funding had more current mentees (mean of 8.3 vs 5.1, respectively; P<.001). Full professors had more current mentees than associate or assistant professors (8.0 vs 5.9 vs 2.4, respectively; P=.005). Ninety-four (85%) mentors had experience comentoring, and two-thirds of mentors had experience mentoring from a distance. Although most mentors found long-distance mentoring to be less demanding, most also said it is less effective for the mentee and is personally less fulfilling. CONCLUSIONS: Mentors in GIM appear to be close to their mentorship capacity, and the majority lack funding for mentorship. Comentoring and long-distance mentoring are common. PMID- 16307627 TI - Best practices for elderly hip fracture patients. A systematic overview of the evidence. AB - OBJECTIVES: To determine evidence-based best practices for elderly hip fracture patients from the time of hospital admission to 6 months postfracture. DATA SOURCES: MEDLINE, Cochrane Library, CINAHL, Embase, PEDro, Ageline, NARIC, and CIRRIE databases were searched for potentially eligible articles published between 1985 and 2004. REVIEW METHODS: Two independent reviewers determined studies appropriate for inclusion using standardized selection criteria, extracted data, evaluated internal validity, and then rated studies according to levels of evidence. Only Level 1 or 2 evidence was included in our summary of clinical recommendations. RESULTS: Spinal anesthesia, pressure-relieving mattresses, perioperative antibiotics, and deep vein thromboses prophylaxes had consistent evidence of benefit. Routine preoperative traction was not associated with any benefits and should be abandoned. Types of surgical management, postoperative wound drainage, and even "multidisciplinary" care, lacked sufficient evidence to determine either benefit or harm. There was little evidence to either determine best subacute rehabilitation practices or to direct ongoing medical issues (e.g., nutrition). Studies conducted during the subacute recovery period were heterogeneous in terms of treatment settings, interventions, and outcomes studied and had no clear evidence for best treatment practices. CONCLUSIONS: The evidence for perioperative practices is relatively robust and evidence-based perioperative treatment guidelines can be easily established. Conversely, more evidence is required to better guide the care of elderly patients with hip fracture during the subacute recovery period and convalescence. PMID- 16307628 TI - The impact of clinical trials on the use of hormone replacement therapy. A population-based study. AB - BACKGROUND: The last 5 years of trial data demonstrate the ineffectiveness of hormone replacement therapy (HRT). The impact of these trials on age-specific HRT use, HRT discontinuation, and regional HRT variation has not been evaluated extensively. OBJECTIVE: To characterize the relation between HRT trial dissemination and age-specific HRT use, HRT discontinuation, and regional HRT variation before and after the trials' publication. DESIGN: Using the Medco Health database, we analyzed HRT prescription filling, discontinuation, and regional variation among women > or =55 years from May 1998 to May 2003. MEASUREMENTS AND MAIN RESULTS: Approximately 340,000 women were eligible for Medco benefits each month. Within 3 months of the Women's Health Initiative (WHI), HRT prescriptions declined from 12.5% to 9.4%, P< or =.0001. When stratified by age, a statistically significant decline in HRT post-WHI occurred in all age groups, with the biggest decline among women > or =55 to 64 (18% to 11%, P< or =.0001). Among HRT users, we found statistically significant increases in discontinuation in 2002 (67%) compared with 2001 (53%, P<.0001). Prior to the WHI there was substantial regional variation in HRT use, with the West South Central and mid-Atlantic having the highest and lowest proportions, respectively (19% vs 6%, P< or =.0001). Despite a relative decline in HRT use of 25% to 42% across all regions, substantial geographic variation remained. CONCLUSIONS: Hormone replacement therapy use decreased significantly immediately post-WHI, suggesting that trial results can have a rapid effect on practice. Marked regional variation in HRT use persisted after the WHI, suggesting that local practice patterns exert a strong effect on clinical behavior even after new evidence is available. PMID- 16307629 TI - The outcomes among patients presenting in primary care with a physical symptom at 5 years. AB - BACKGROUND: Symptoms are common and often remain medically unexplained. OBJECTIVE: To assess 5-year symptom outcomes, determine how often symptoms remain unexplained and assess their relationship with mental disorders. DESIGN: Prospective cohort study. PARTICIPANTS: Five hundred consecutive patients presenting to a medicine clinic with physical symptoms. MEASUREMENTS PREVISIT: Mental disorders, symptom characteristics, stress, expectations, illness worry, and functional status. Postvisit (immediately, 2 weeks, 3 months, 5 years): unmet expectations, satisfaction, symptom outcome, functional status, and stress. RESULTS: While most subjects (81%) experienced symptom improvement by 5 years, resolution rates were lower (56%), with 35% of symptoms remained medically unexplained. Most patients with medically unexplained symptoms (MUS) did not have a mental disorder. Mood or anxiety disorders were not associated with MUS (relative risks [RR]: 0.94, 95% confidence interval [CI]: 0.79 to 1.13), or with lower rates of symptom improvement (RR: 1.14, 95% CI: 0.60 to 2.2). In contrast, most patients with somatoform disorders had MUS and were unlikely to improve. Worse functioning (RR: 0.95, 95% CI: 0.91 to 0.99), longer duration of symptom at presentation (RR: 0.5, 95% CI: 0.28 to 0.87), illness worry at presentation (RR: 0.56, 95% CI: 0.35 to 0.89), or lack of resolution by 3 months (RR: 0.47, 95% CI: 0.26 to 0.86) reduced the likelihood of symptom improvement at 5 years. CONCLUSIONS: More than half of patients presenting with a physical symptom resolve by 5 years, while a third remain medically unexplained. Most patients whose symptom remained unexplained had no mental disorder. While mood and anxiety disorders were not associated with MUS or worse outcomes, most patients with somatoform disorders had MUS and were unlikely to improve. PMID- 16307630 TI - Buprenorphine retention in primary care. AB - BACKGROUND: This study assesses the rate and predictors of treatment retention for primary care patients with opioid dependence-prescribed buprenorphine, a long acting partial opioid agonist. METHODS: Observational cohort study of patients prescribed buprenorphine/naloxone and followed for 6 months in the period after the adoption of buprenophine/naloxone by a primary care practice in Rhode Island. Practice policy precluded patient discharges due to continuing drug use. RESULTS: Patients (n=41) had a mean duration of opioid use of 15.7 years and most had a history of heroin use (63.4%). Thirty-nine percent of patients transferred from methadone maintenance. At 24 weeks, 59% remained in treatment. Nearly half of dropouts occurred in the first 30 days. Participants with opiate-positive toxicologies at week 1 were more likely to drop out of the program (P<.01) and had a significantly shorter retention time (P<.01) on average. Among other drug use and drug treatment variables, employment and addiction counseling during treatment were significantly associated with treatment retention (P=.03). CONCLUSION: Retention rates in a real world, primary care-based buprenorphine maintenance practice reflect those reported in clinical trials. Abstinence during the first week of treatment and receipt of counseling were critical to patient retention. PMID- 16307631 TI - Brief report: Resident recognition of low literacy as a risk factor in hospital readmission. AB - BACKGROUND: Low literacy is associated with poor self-management of disease and increased hospitalization, yet few studies have explored the extent to which physicians consider literacy in their patient care. OBJECTIVE: To examine trainee recognition of low literacy as a potential factor in patient adherence and hospital readmission. DESIGN AND PARTICIPANTS: Randomized study of 98 Internal Medicine residents and medical students. Trainees reviewed a case history and completed a questionnaire pertaining to a fictional patient's hospital readmission. Case version A contained clues to suggest limited patient literacy skills, while version B did not. Responses were reviewed for mention of low literacy and educational strategies recommended for low-literate patients. RESULTS: Few trainees raised the possibility of low patient literacy, even when provided clues (25% in Group A vs 4% in Group B, P=.003). Furthermore, while most trainees listed patient education as an important means of preventing another readmission, only 16% suggested using a strategy recommended for low-literate adults. CONCLUSION: Few trainees recognized low literacy as a potential factor in patient nonadherence and hospital readmission, and few recommended low-literate educational strategies. Medical residents and students may benefit from additional training in the recognition and counseling of low-literate patients. PMID- 16307632 TI - Health services research in the privacy age. AB - BACKGROUND: Strengthened regulations concerning privacy of health information are affecting large-scale health outcomes research. OBJECTIVE: To create a data collection system that would facilitate outcomes research, avoid selection bias, and fulfill obligations to protect privacy. DESIGN: We created a web-based system that uses touch-screen computer technology for longitudinal collection of data. The system provides access to information in deidentified form, enables it to be linked to health services and outcomes data, and allows patients to join a research registry project (RRP) and be placed on a prospective subject list (PSL). PARTICIPANTS, MEASUREMENTS, AND RESULTS: Pilot testing in 86 consecutive patients who were seen at a large, urban, university-based general medicine practice and had a mean age of 50 years showed that 81 patients had no difficulty, 5 had some difficulty, and none had considerable difficulty using the computer technology to complete a health survey. No patients refused to complete the survey and all patients completed the entire survey. Forty-seven (55%) joined the RRP and 42 of these 47 (89%) joined the PSL. RRP participants were less likely than RRP nonparticipants to be divorced or widowed (P=.03) and less likely to have hypertension (P=.03) but had no other significant differences in sociodemographic or clinical characteristics. PSL participants did not differ from PSL nonparticipants. CONCLUSIONS: The new system ensures privacy and appears to facilitate research recruitment and avoid selection bias. PMID- 16307633 TI - Interpreter services, language concordance, and health care quality. Experiences of Asian Americans with limited English proficiency. AB - BACKGROUND: Patients with limited English proficiency (LEP) have more difficulty communicating with health care providers and are less satisfied with their care than others. Both interpreter- and language-concordant clinicians may help overcome these problems but few studies have compared these approaches. OBJECTIVE: To compare self-reported communication and visit ratings for LEP Asian immigrants whose visits involve either a clinic interpreter or a clinician speaking their native language. DESIGN: Cross-sectional survey-response rate 74%. PATIENTS: Two thousand seven hundred and fifteen LEP Chinese and Vietnamese immigrant adults who received care at 11 community-based health centers across the U.S. MEASUREMENTS: Five self-reported communication measures and overall rating of care. RESULTS: Patients who used interpreters were more likely than language-concordant patients to report having questions about their care (30.1% vs 20.9%, P<.001) or about mental health (25.3% vs 18.2%, P=.005) they wanted to ask but did not. They did not differ significantly in their response to 3 other communication measures or their likelihood of rating the health care received as "excellent" or "very good" (51.7% vs 50.9%, P=.8). Patients who rated their interpreters highly ("excellent" or "very good") were more likely to rate the health care they received highly (adjusted odds ratio 4.8, 95% confidence interval, 2.3 to 10.1). CONCLUSIONS: Assessments of communication and health care quality for outpatient visits are similar for LEP Asian immigrants who use interpreters and those whose clinicians speak their language. However, interpreter use may compromise certain aspects of communication. The perceived quality of the interpreter is strongly associated with patients' assessments of quality of care overall. PMID- 16307634 TI - The doctor will see you shortly. The ethical significance of time for the patient physician relationship. AB - Many physicians and health care leaders express concern about the amount of time available for clinical practice. While debates rage on about how much time is truly available, the perception that time is inadequate is now pervasive. This perception has ethical significance, because it may cause clinicians to forego activities and behaviors that promote important aspects of the patient-physician relationship, to shortcut shared decision making, and to fall short of obligations to act as patient advocates. Furthermore, perceived time constraints can hinder the just distribution of physician time. Although creating more time in the clinical encounter would certainly address these ethical concerns, specific strategies-many of which do not take significantly more time-can effectively change the perception that time is inadequate. These approaches are critical for clinicians and health systems to maintain their ethical commitments and simultaneously deal with the realities of time. PMID- 16307635 TI - Use of critical incident reports in medical education. A perspective. AB - Critical incident reports are now being widely used in medical education. They are short narrative accounts focusing on the most important professional experiences of medical students, residents, and other learners. As such, critical incident reports are ideally suited for addressing values and attitudes, and teaching professional development. This manuscript describes critical incident reports and gives examples of their use, provides a theoretical underpinning that explains their effectiveness, and describes the educational impacts of critical incident reports and similar methods that use reflective learning. The author recommends critical incident reports as an especially effective means to address learners' most deeply held values and attitudes in the context of their professional experiences. PMID- 16307636 TI - Colorectal cancer screening. The time is now! PMID- 16307637 TI - Weakness in numbers. The challenge of numeracy in health care. PMID- 16307638 TI - The waiting room. PMID- 16307639 TI - A bag of humility. PMID- 16307641 TI - Sentinel node and survival in melanoma. PMID- 16307642 TI - The dual action of ozone on the skin. AB - The aim of this brief review is to summarize the recent literature on the effect of ozone (O3) on cutaneous tissues. Recently it has been reported that a chronic contact with O3 can be deleterious for the skin. Our group and others have shown a progressive depletion of antioxidant content in the stratum corneum and this can then lead to a cascade of effects resulting in an active cellular response in the deeper layers of the skin. Using an in vivo model we have shown an increase of proliferative, adaptive and proinflammatory cutaneous tissue responses. On the other hand the well known activity of O3 as a potent disinfectant and oxygen (O2) donor has been also studied for therapeutic use. Two approaches have been described. The first consists of a quasi-total body exposure in a thermostatically controlled cabin. This treatment has proved to be useful in patients with chronic limb ischaemia. The second approach is based on the topical application of ozonated olive oil in several kinds of skin infection (from soreness to diabetic ulcers, burns, traumatic and surgical wounds, abscesses and skin reactions after radiotherapy). We and other authors have observed a striking cleansing effect with improved oxygenation and enhanced healing of these conditions. It is now clear that, on the skin, O3, like other drugs, poisons and radiation, can display either a damaging effect from a long exposure or a beneficial effect after a brief exposure to O2 and O3 or to the application of ozonated oil to chronic wounds. PMID- 16307643 TI - Long-term culture of adult murine epidermal keratinocytes. AB - BACKGROUND: Long-term cultures of epidermal cells from mouse skin have been notoriously difficult to establish. OBJECTIVES: To develop a modified serum-free medium and technique for long-term culture of adult mouse epidermal keratinocytes. METHODS: Epidermal cells from trypsin-treated adult mouse dorsal and ventral skin were grown on type I collagen-coated dishes without feeder layers in a serum-free medium supplemented with only 10 ng mL(-1) epidermal growth factor (EGF) and 10(-10) mol L(-1) cholera toxin (CT). RESULTS: After removing coexisting fibroblasts several times, we were able to obtain almost pure basal epidermal keratinocytes. Our technique supports sustained multiplication of mouse basal keratinocytes for more than 100 population doublings, and they retained the capacity to undergo terminal differentiation when given the appropriate stimulus. The epithelial nature of these cultivated cells was demonstrated both by phase-contrast microscopy and by immunostaining with antikeratin antibodies. EGF and CT, which have been reported to accelerate the cellular growth rate, were essential for successful long-term cultivation during multiple passages. CONCLUSIONS: Our technique is very simple. It provides a useful and suitable model for investigations of growth, differentiation and skin remodelling in vitro. PMID- 16307644 TI - Induction of toll-like receptors by Propionibacterium acnes. AB - BACKGROUND: The bacterium Propionibacterium acnes is involved in the induction and maintenance of the inflammatory phase of acne. Recent studies have found that keratinocytes express toll-like receptors (TLRs) implicated in immediate immunity. No studies have, to date, been carried out on the action of P. acnes upon TLR activation in keratinocytes. OBJECTIVES: Focusing on the inflammatory phase of acne, to clarify the role of P. acnes in immediate immunity by inducing expression of TLR-2 and TLR-4 by keratinocytes. We also studied how the secretion and expression of matrix metalloproteinase (MMP)-9 is induced by P. acnes. METHODS: The work was carried out on two levels: in vivo with the study of the expression of TLR-2 and TLR-4 proteins in biopsies of acne lesions and in vitro on cultured keratinocyte monolayers to study the modulating effects of P. acnes on the expression of TLR-2 and TLR-4 and also on the expression and secretion of MMP-9. RESULTS: Our findings reveal that in vivo TLR-2 and TLR-4 expression is increased in the epidermis of acne lesions. In vitro, an increase in TLR-2 and TLR-4 expression by human keratinocytes occurred in the first hours of incubation with bacterial fractions as well as an increase of the expression and secretion by the keratinocytes of MMP-9, which plays a role in inflammation. CONCLUSIONS: This work demonstrates that P. acnes induces TLR expression and that this mechanism could play an essential role in acne-linked inflammation. These receptors could be involved notably in acute acne. PMID- 16307645 TI - Response of the hypothalamic-pituitary-adrenal axis to psychological stress in patients with psoriasis. AB - BACKGROUND: Psoriasis may, in some patients, be triggered and/or exacerbated by stress. OBJECTIVES: As activation of the hypothalamic-pituitary-adrenal (HPA) axis is critical to a successful stress response we investigated this in patients with psoriasis. METHODS: Forty patients with chronic plaque psoriasis and 40 age matched normal controls experienced three randomly presented acute psychological stressors (cognitive, emotional and social). Serial serum cortisol, pulse rate and blood pressure assessments were undertaken at baseline and following each of the stressors. Salivary cortisol samples were collected at 09.00 h on the day of testing. RESULTS: In control subjects there was a significant (r = 0.38; P < 0.05) correlation between pulse rate and serum cortisol level following the social performance stressor; this was not evident in the psoriasis group (r = 0.07; not significant). Patients who believed that their psoriasis was highly stress responsive had significantly lower salivary cortisol levels at baseline (P < 0.01) and lower serum cortisol levels following the social performance stressor (P = 0.016) than patients with nonstress-responsive disease who believed that stress had no impact. In contrast, there was no difference between the groups for change in pulse rate poststressor. CONCLUSIONS: This study shows that patients with psoriasis, and in particular those whose disease appears to be stress responsive, exhibit an altered HPA response to acute social stress. The implication is that such patients may perhaps be primed to flares of their psoriasis. Whether this is genetically predetermined and/or a consequence of the distress of living with psoriasis remains to be determined. PMID- 16307646 TI - CDKN2A mutations in Scottish families with cutaneous melanoma: results from 32 newly identified families. AB - BACKGROUND: Up to 5% of patients with melanoma have a family history of a first degree relative also being affected. OBJECTIVES: To study such families for germline mutations, to help clarify the gene-environment interaction in melanoma aetiology. METHODS: Thirty-two families in Scotland with melanoma in two or more first-degree relatives are reported for the first time. Peripheral blood DNA was extracted, and denaturing high-performance liquid chromatography analysis performed on exons 1alpha and 2 of the CDKN2A gene and their splice junctions. The coding sequences and splice junctions of these exons were sequenced in all samples as confirmation of the chromatographic pattern observed. RESULTS: Seven of the 32 melanoma families (22%) have CDKN2A mutations. One mutation, H83N, which has not previously been described in melanoma families, was found in one family. In addition, two families have R112G mutations, one family has a G67R mutation, one has an exon 1alpha 24-bp duplication where bases 9-32 are duplicated between bases 32 and 33, and two families have M53I mutations, bringing the total of known Scottish families with the M53I mutation to six. CONCLUSIONS: This study brings the total of Scottish families investigated for germline mutations to 48, and strongly suggests that the M53I mutation originated in Scotland. PMID- 16307647 TI - Efficacy of oral isotretinoin in the control of skin and nasal colonization by antibiotic-resistant propionibacteria in patients with acne. AB - BACKGROUND: Skin colonization by antibiotic-resistant propionibacteria is commonplace among acne patients globally. Increasing attention is now being paid to how resistance rates might be reduced to preserve the future efficacy of antibiotics, especially erythromycin and clindamycin in acne therapy. OBJECTIVE: To assess the efficacy of oral isotretinoin in the control of antibiotic resistant propionibacteria. METHODS: Acne patients (72 in the U.K., 62 in the U.S.A.) colonized with high numbers of antibiotic-resistant propionibacteria were sampled before, during and 12 weeks after oral isotretinoin therapy. Propionibacterial samples were collected from five acne-prone skin surface sites using a detergent scrub method and from the anterior nares using moistened swabs. Total and antibiotic-resistant propionibacteria were enumerated by viable counting on media with and without selective antibiotics. RESULTS: After 16 weeks of oral isotretinoin therapy, mean population densities of viable propionibacteria and variants resistant to erythromycin, clindamycin or tetracycline had fallen by more than 90% at all skin sites and in the nares. The sole exception was a smaller reduction in tetracycline-resistant strains on the lower back. In general, greater reductions were observed on skin than in the nares. By the end of the treatment period only three patients (all in Philadelphia) yielded no antibiotic-resistant strains from any site. Post treatment, propionibacterial counts remained well below pretreatment levels but had begun to recover on the face and in the nares. The recovering propionibacterial population included both susceptible and resistant strains. Changes during and post-treatment at the two centres were similar but not identical. CONCLUSIONS: Oral isotretinoin effectively reduced skin and nasal colonization by antibiotic-resistant propionibacteria. However, viable populations of resistant isolates persisted post-treatment at multiple sites. Novel methods are required to eradicate antibiotic-resistant propionibacteria completely, especially from the nasal reservoir. PMID- 16307648 TI - Sentinel node biopsy in melanoma delays recurrence but does not change melanoma related survival: a retrospective analysis of 673 patients. AB - BACKGROUND: The status of the sentinel lymph node (SLN) is an important parameter to predict the prognosis of melanoma patients but it is a matter of debate if removal of micrometastases by SLN biopsy (SLNB) influences the prognosis of melanoma patients. OBJECTIVES: We sought to investigate the impact of SLNB in melanoma patients with regard to recurrence-free survival, overall survival and metastatic pathways. PATIENTS AND METHODS: We studied, retrospectively, 673 melanoma patients with a primary melanoma (tumour thickness > or = 1 mm) and without clinical evidence of metastases at the time of melanoma diagnosis. In 377 patients the melanoma was removed without SLNB between January 1995 and March 2000 (pre-SLNB group). In 296 patients the melanoma was removed with SLNB between April 2000 and March 2003 (SLNB group). Otherwise, both groups received identical surgical treatment of the primary melanoma and initial staging procedures performed by the same team of physicians. Follow-up recommendations were also identical in both groups. RESULTS: Both groups showed no significant differences with regard to characteristics of the primary melanoma, sex and age. By Kaplan Meier analyses, melanoma-related overall survival was comparable in both groups. However, recurrence-free survival was increased in pre-SLNB patients due to significantly fewer regional lymph node metastases, whereas frequencies of locoregional cutaneous and distant metastases were comparable in both groups. CONCLUSIONS: SLNB advances the detection of regional lymph node metastases and therefore avoids nodal recurrences but does not influence metastatic behaviour of melanoma cells and does not protect patients from melanoma-related death caused by distant metastases. Thus, our retrospective data favour the marker hypothesis for melanoma metastasation. To elucidate further if subgroups of patients benefit from SLNB, prospective randomized studies with long-term follow-up are needed. PMID- 16307649 TI - Less skin irritation from alcohol-based disinfectant than from detergent used for hand disinfection. AB - BACKGROUND: The benefit of alcohol-based disinfectant used on normal skin has been debated. OBJECTIVES: The objective of the present study was to compare the effects of repeated exposure to an alcohol-based disinfectant, a detergent and an alcohol-based disinfectant/detergent alternately for 10 days, including noninvasive measurements in the evaluation. Skin reactivity in irritated skin after a 4-week interval was also evaluated. MATERIALS AND METHODS: Detergent, disinfectant and alternate disinfectant and detergent were applied twice daily every 10 min for 1 h to the ventral upper arms and forearms of 17 healthy volunteers. A control area was included. After 4 weeks an SLS patch was applied to each area. Irritant reactions were quantified with a visual score recording and measurements of transepidermal water loss (TEWL) and skin colour were performed on days 1, 5, 11, 38 and 40. RESULTS: On day 5 the detergent caused a higher visual score than either disinfectant applied alone or alternate disinfectant and detergent, P < 0.05. On day 11 detergent and alternate disinfectant and detergent caused a higher score than disinfectant, P < 0.05. An increased irritant response for detergent compared with disinfectant alone and alternate disinfectant and detergent was confirmed by TEWL evaluations on days 5 and 11, P < 0.001, while no significant difference was found by colour measurement. After 4 weeks, when evaluated by colour measurement, significantly less response to the SLS patch was found on the disinfectant-treated area compared with the control area and detergent area, and a similar trend was found for TEWL, although it was not statistically significant. CONCLUSION: Alcohol based disinfectant caused less visible skin irritation and less skin barrier disruption than the use of detergent. The alternate use of detergent and disinfectant caused less irritation than the use of detergent, and a possible interaction between the two irritants was not indicated. After 4 weeks there was a tendency for decreased skin reactivity in the skin areas which had 4 weeks previously been exposed to disinfectant. PMID- 16307650 TI - Cutaneous lip tumours treated with Mohs micrographic surgery: clinical features and surgical outcome. AB - BACKGROUND: The Australian Mohs micrographic surgery (MMS) database was initiated in 1993 by the Skin and Cancer Foundation Australia (SCFA) with the aim of collecting prospective data, and involved all Mohs surgeons in the country. OBJECTIVES: To present a large series of patients with cutaneous lip tumours treated with MMS in Australia between 1993 and 2002. METHODS: This prospective multicentre case series included all patients with cutaneous lip tumours who were monitored by the SCFA. The main outcome measures were patient demographics, reason for referral, duration of tumour, site, preoperative tumour size and postoperative defect size, recurrences prior to MMS, histological subtypes, perineural invasion and 5-year recurrence after MMS. RESULTS: There were 581 patients (66.1% women and 33.9% men, P < 0.0001) with a mean +/- SD age of 58 +/- 15 years. The upper lip was the most common site involved (81.1%). Basal cell carcinoma (BCC) was diagnosed in 82.3%, squamous cell carcinoma (SCC) in 16.5%, Bowen's disease (BD) in 0.7% and microcystic adnexal carcinoma (MAC) in 0.5% of cases. BCC was more common on the upper lip and in women, whereas SCC was more common on the lower lip and in men (P < 0.0001). Most upper lip tumours occurred in women (75.4%), whereas most lower lip tumours occurred in men (73.6%). SCC was associated with a larger tumour and postoperative defect size compared with the other tumours. The 5-year recurrence for BCC was 3.0%, and there were no cases of recurrence for SCC, BD or MAC. CONCLUSIONS: BCC was the most common cutaneous lip tumour managed by MMS, and was significantly more common on the upper lip and in women. The low 5-year recurrence rate emphasizes the importance of margin controlled excision. PMID- 16307651 TI - Diagnosis of nail psoriasis: importance of biopsy and histopathology. AB - BACKGROUND: Involvement of the nail is quite common in psoriasis and at times may be the sole diagnostic clue. However, the histopathology of nail psoriasis has not been adequately evaluated. A confirmation of the diagnosis is required in cases suspected to have nail psoriasis in order to plan long-term therapy. OBJECTIVES: To assess the diagnostic significance and safety profile of nail biopsy in cases with nail psoriasis. METHODS: Clinical and mycological features were studied in 42 patients with nail psoriasis. Of these, 22 patients gave consent for nail biopsies to be taken and the histopathological changes were assessed. RESULTS: Males were affected more commonly (57%) with a peak incidence in the age group of 10-20 years (29%). Distal onycholysis, discoloration of nail plate, subungual hyperkeratosis and fine nail pitting were the predominant clinical features. In the 22 biopsies done, hyperkeratosis with parakeratosis (91%) was found to be the most common and hypergranulosis was the least common histological finding (36%). Clinicohistological correlation was possible only in 55% cases. Periodic acid-Schiff (PAS) staining was done for all biopsies. CONCLUSIONS: Histopathological examination of nails is a valuable diagnostic aid, especially in the absence of skin lesions. Examination of the PAS-stained sections is necessary before making a histological diagnosis of nail psoriasis because onychomycosis and psoriasis may show similar histology. PMID- 16307652 TI - Distribution and colocalization of markers for proliferation, invasion, motility and neoangiogenesis in benign melanocytic naevi and malignant melanomas. AB - BACKGROUND: Melanomas are heterogeneous tumours, and differentiation from other melanocytic lesions may cause problems. It may be possible that the distribution and/or colocalization pattern of different markers in the lesions can enable a more accurate diagnosis of melanocytic tumours. OBJECTIVES: To test this hypothesis, melanocytic naevi, primary melanomas and metastases were investigated. METHODS: The distribution and colocalization of markers for proliferation, invasion, angiogenesis and motility of the tumour cells were investigated using antibodies directed against actin, cathepsin B (CatB), transforming growth factor-beta, vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen/Ki-67 and basic fibroblast growth factor (FGF 2). In addition, melanoma markers (HMB-45 and Melan-A) and proteins unrelated to melanoma progression [epidermal growth factor (EGF) and cathepsin H] were investigated. RESULTS: Malignant melanomas tended to express more markers of malignancy compared with melanocytic naevi, and the differences were statistically significant for EGF and actin immunoreactivity: melanocytic naevi displayed clear EGF labelling more often (60% vs. 5%) and melanomas showed more intense actin labelling (70% vs. 0%). HMB-45+ cells to a large extent also stained with antibodies to CatB but not to EGF or actin; EGF-, FGF-2- and VEGF immunoreactive cells were predominantly HMB-45-. Similar combinations were observed in melanocytic naevi and in melanomas. CONCLUSIONS: Labelling with EGF may improve the differential diagnosis of melanocytic neoplasias. However, we did not detect a clear-cut increase of markers of malignancy in melanoma. Cells expressing multiple malignancy markers were also found in some melanocytic naevi; this may confirm the dormant potential of melanocytic naevi for melanoma development. PMID- 16307653 TI - Cyclin A and beta-catenin expression in actinic keratosis, Bowen's disease and invasive squamous cell carcinoma of the skin. AB - BACKGROUND: Actinic keratosis (AK) has been defined as a precancerous lesion or an early phase in the evolution of squamous cell carcinoma (SCC) and histological changes seen in the individual cells of an AK are indistinguishable from those seen in SCC, which invade the dermis. Cyclin A is an increasingly utilized proliferation marker that has functions in both S phase (DNA replication) and initiation of mitosis, whereas alterations of beta-catenin, the molecule involved in cell-cell adhesion and in signalling transduction, could promote invasive and proliferative capacities of malignant tumours. OBJECTIVES: To determine cyclin A and beta-catenin expression pattern in cutaneous SCC and in in situ lesions classified as keratinocytic intraepidermal neoplasia (KIN) and, using traditional terms, as AK and Bowen's disease (BD), and to analyse it in relation to SCC differentiation, diameter and thickness. METHODS: Immunohistochemical staining was performed on 110 formalin-fixed paraffin-embedded tissue samples with the streptavidin-biotin technique using antibodies to cyclin A and beta-catenin. On histological examination, 53 lesions were diagnosed as AK, 16 as BD and 41 as SCC 11 well differentiated (WD), 16 moderately differentiated (MD) and 14 poorly differentiated (PD). Using KIN classification, 22 lesions were KIN1, 23 were KIN2 and 24 were KIN3. For cyclin A, distribution and labelling index (LI), and for beta-catenin, level of membranous staining and presence of aberrant (nuclear/cytoplasmic) localization were examined. RESULTS: Diffuse cyclin A presence was observed more frequently in BD than in AK (P < 0.0001) or SCC (P = 0.0002), and in SCC-PD compared with SCC-WD (P < 0.0001) or SCC-MD (P = 0.0003). Differences between KIN3 and KIN2, as well as KIN3 and KIN1 lesions, were statistically significant (P < 0.0001), and the same result appeared when KIN1 and KIN2 cases were grouped and compared with those of KIN3 (P < 0.0001). Cyclin A LI was significantly lower (P < 0.05) in AK than in BD or SCC, but no difference between BD and SCC was found, and LI in BD was even higher than in SCC WD or SCC-MD, while analysis regarding SCC differentiation and KIN classification revealed the same correlation as for the cyclin A distribution. Reduced or absent beta-catenin membranous staining was found in 90 cases (81.8%), more often in SCC than in AK (P = 0.03) or in AK and BD grouped together (P = 0.02). There was no statistical difference between SCCs of various level of differentiation, or between different KIN grades. Diffuse loss of membranous beta-catenin staining showed 36 lesions (32.7%), more frequently SCC than AK (P = 0.003) or AK and BD grouped (P = 0.006), as well as SCC-PD compared with SCC-WD (P = 0.01) and SCC-MD (P = 0.03), whereas all KIN comparisons remained nonsignificant. Aberrant beta catenin cellular localization demonstrated 28 lesions (25.5%), most often in the basal or peripheral parts and in the lesions with diffuse beta-catenin loss (P = 0.009), but revealed no correlation with the histological type, SCC level of differentiation or KIN grades. Diffuse loss of membranous beta-catenin staining was found to be significantly more frequent in SCC thicker than 4 mm (P = 0.03), while all other comparisons between cyclin A or beta-catenin with the tumour size remained nonsignificant. Cyclin A LI was higher in cases with diffuse loss of membranous staining (P = 0.001) or with aberrant cellular localization of beta catenin (P = 0.002). CONCLUSIONS: Cyclin A LI showed greater difference between AK and BD than between BD and SCC, suggesting that increased proliferation (measured by cyclin A LI) characterizes progression of in situ lesions from AK to BD, whereas reduced beta-catenin expression separates more clearly SCC from the in situ lesions. Diffuse pattern of loss of membranous beta-catenin staining correlated better with the type of lesion, SCC differentiation and tumour size than reduced expression in general or aberrant cellular localization of beta catenin. KIN classification does not seem to be supported by our findings, except when KIN1 and KIN2 lesions (in situ, partial thickness) are grouped. PMID- 16307654 TI - Comorbidity of rosacea and depression: an analysis of the National Ambulatory Medical Care Survey and National Hospital Ambulatory Care Survey--Outpatient Department data collected by the U.S. National Center for Health Statistics from 1995 to 2002. AB - BACKGROUND: Psychogenic factors have been considered to be important in the exacerbation and possibly the onset of rosacea. However, there are very few studies that have reported conclusive findings. OBJECTIVES: To examine the association between rosacea and major depressive disease, a common and usually treatable psychiatric disorder. METHODS: Data from 1995 to 2002, collected by the National Ambulatory Medical Care Survey and the outpatient component of the National Hospital Ambulatory Care Survey, which are both nationally representative surveys of healthcare visits in the U.S.A., were studied. The basic sampling unit in both surveys is the patient visit or encounter. A "Rosacea" variable was created by grouping all rosacea (ICD-9-CM code 695.3) visits and a "Depression" variable was created by grouping the patient visits related to major depressive disorder (ICD-9-CM codes 296.2, 296.3 and 311). As alcohol abuse has been implicated in rosacea, and alcohol can confound symptoms of depression, an "Alcohol" variable was created by grouping all ICD-9-CM codes related to alcohol dependence and abuse (codes 303, 303.0, 303.9 and 305.0). All analyses were conducted using the Complex Samples module of SPSS version 13, to account for the multistage probability sampling design used to collect the data. RESULTS: The weighted data were representative of over 608 million dermatology visits between 1995 and 2002. Logistic regression analysis using "Rosacea" as the dependent variable and age, sex, "Alcohol" and "Depression" as independent variables revealed that the odds ratio for depressive disease in the rosacea group was 4.81 (95% confidence interval 1.39-16.62). The association between "Alcohol" and "Rosacea" was not significant. CONCLUSIONS: The comorbidity between major depressive disease and rosacea may have important clinical implications. Alcohol abuse does not appear to play a significant role in this association. PMID- 16307655 TI - ATX-S10(Na)-photodynamic therapy is less carcinogenic for mouse skin compared with ultraviolet B irradiation. AB - BACKGROUND: Photodynamic therapy (PDT) is available for the treatment of various skin tumours and other skin diseases. Ultraviolet (UV) irradiation induces DNA damage, cyclobutane pyrimidine dimers (CPD) (6-4) photoproducts (6-4PP) and 8 hydroxy-2'-deoxyguanosine (8-OHdG), all of which are carcinogenic for the skin. However, effects of PDT on DNA damage and carcinogenesis are unclear. OBJECTIVES: To compare the production of photoproducts and the induction of skin tumours in mouse epidermis treated with UVB or PDT. METHODS: We performed UVB irradiation or ATX-S10(Na)-PDT on the skin of 20 hairless mice, in each case, and analysed DNA damage and tumour induction. RESULTS: After a single irradiation of UVB on mouse skin, CPD, 6-4PP and 8-OHdG were detected in the nuclei of keratinocytes. In contrast, PDT-treated mouse keratinocytes showed induction of 8-OHdG, but not of CPD or 6-4PP. Skin tumours induced by UVB irradiation (3 kJ m(-2) three times weekly) were observed following 15 weeks of irradiation (mean +/- SEM tumour incidence 3.2 +/- 1.8%; tumour number 3.2 +/- 1.6 per mouse) and increased depending on irradiation times and doses. Following 30 weeks of UVB irradiation (3 kJ m(-2) three times weekly), mean +/- SEM tumour incidence and tumour number were 28.7 +/- 4.8% and 14.2 +/- 2.8% per mouse, respectively. Although skin tumours were also detected in PDT-treated mouse skin following 80 weeks of treatment (mean +/- SEM tumour incidence 9.1 +/- 1.8%; tumour number 12.2 +/- 2.3 per mouse), the number of tumours was not statistically different from untreated mouse skin (mean +/- SEM tumour incidence 4.1 +/- 3.8%; tumour number 5.2 +/- 3.3 per mouse). CONCLUSIONS: PDT induced 8-OHDG but not CPD or 6-4PP, and was shown to be a relatively safe modality following multiple applications to mouse skin. PMID- 16307656 TI - Can St John's wort (hypericin) ingestion enhance the erythemal response during high-dose ultraviolet A1 therapy? AB - BACKGROUND: St John's wort (SJW) is widely used as a treatment for depression. A phototoxic reaction, due to its content of hypericin, can occur in animals and in cell culture, and has been reported in humans. Hypericin displays absorption within the ultraviolet (UV) A1 spectrum and there may therefore be a potential for phototoxicity if taken during high-dose UVA1 therapy. OBJECTIVES: To assess the phototoxicity risk of SJW ingestion. METHODS: Eleven adult volunteers of skin types I and II were exposed to a geometric dose series of UVA1 irradiation from a high-output source (Dermalight Ultra 1; Dr Honle, Martinsreid, Germany; irradiance 70-77 mW cm(-2)) on the photoprotected lower back skin at eight 1.5 cm(2) test areas. Irradiation was carried out at baseline and after 10 days of SJW extract 1020 mg (equivalent to 3000 microg of hypericin) daily. Four, 8, 24 and 48 h after each exposure, the minimal erythema dose (MED) and the presence or absence of pigmentation were recorded visually and erythema was assessed objectively with an erythema meter. RESULTS: The median MED and D(0.025), an objective measure of MED, were lower at all time-points after SJW ingestion. The visual erythemal peak (lowest median MED), which was seen at 8 h postirradiation, was lower after SJW (median 14 J cm(-2), range 10-56) than at baseline (median 20 J cm(-2), range 14-56) (P = 0.047). Similarly, the median D(0.025) at 8 h postirradiation was lower after SJW (median 22.0 J cm(-2), range 15.2-53.9) than at baseline (median 33.7 J cm(-2), range 22.9-136.0) (P = 0.014). The MED and D(0.025) were also significantly different at the 48-h and 4-h time-points, respectively. Significance was not reached at the 24-h time-point. Median intensity of postirradiation erythema increased at all time-points after ingestion of SJW. Despite these differences, the maximum slope of the dose response curve was not increased after SJW ingestion. CONCLUSIONS: These data suggest that SJW extract has the potential to lower the erythemal threshold to UVA1 irradiation in a significant proportion of individuals and highlight the importance of ascertaining a full drug history, including herbal remedies, before initiating UVA1 phototherapy. PMID- 16307657 TI - Patient-reported outcomes of psoriasis improvement with etanercept therapy: results of a randomized phase III trial. AB - BACKGROUND: Etanercept, a soluble tumour necrosis factor receptor, lessens the severity of psoriasis as measured by physician-reported clinical outcomes. Equally important is the patient perspective on the effect of etanercept therapy on daily life. OBJECTIVES: To assess patient-reported outcomes (PROs) in patients with psoriasis receiving etanercept therapy. METHODS: In this multinational, randomized, phase III trial, patients with psoriasis received placebo (n = 193), etanercept 50 mg per week (n = 196) or etanercept 50 mg twice weekly (n = 194) during the initial 12-week, double-blind period. Thereafter, all patients received open-label etanercept (50 mg per week). The following PROs were assessed: Dermatology Life Quality Index (DLQI), Short Form-36 Health Survey (SF 36), patient rating of pruritus, and patient global assessment of psoriasis. RESULTS: At week 12, DLQI total score improved by 65-70% in patients receiving etanercept compared with 6% in patients receiving placebo (P < 0.0001), and improvement in DLQI was clinically meaningful (> or = 5-point improvement or 0 score) for 72-77% of patients receiving etanercept therapy. All DLQI and SF-36 subscales and the SF-36 physical and mental component summary scores demonstrated significantly greater improvement with etanercept therapy than with placebo, illustrating that etanercept benefits patients with psoriasis across multiple domains that contribute to health-related quality of life. With etanercept therapy, distributions of patient ratings of pruritus and global assessment of disease shifted from moderate to severe (baseline) to minimal to good (week 12). Etanercept-induced benefits of PROs were maintained for patients who reduced their dose after 12 weeks. CONCLUSIONS: Etanercept therapy improves PROs in patients with psoriasis and makes a meaningful difference to their lives. These results support the efficacy profile of physician-reported clinical measures while providing a more complete understanding of the benefits experienced by patients with psoriasis treated with etanercept. PMID- 16307658 TI - hK5 and hK7, two serine proteinases abundant in human skin, are inhibited by LEKTI domain 6. AB - BACKGROUND: Several skin diseases and atopic disorders including Netherton syndrome and atopic dermatitis have been associated with mutations and deviations of expression of SPINK5, the gene encoding the human 15-domain serine proteinase inhibitor LEKTI. The biochemical mechanisms underlying this phenomenon have not yet been fully clarified. OBJECTIVES: To identify target proteinases of LEKTI important for processes of desquamation and inflammation of the skin which will enable the development of specific drugs. METHODS: The inhibitory activities of LEKTI domains 6 and 15 were tested on a number of commercially available serine proteinases and also on the purified kallikreins hK5 and hK7. In addition, recombinant hK5 was used. RESULTS: LEKTI domain 6 is a potent inhibitor of hK5 and hK7, whereas LEKTI domain 15 exhibits inhibitory activity on plasmin. hK5 and hK7 in particular are relevant to skin disorders. CONCLUSIONS: The inhibition of hK5 and hK7 by LEKTI domain 6 indicates an important regulatory role of LEKTI in processes of skin desquamation and inflammation, which may explain the severe pathological symptoms associated with abnormalities of SPINK5 and/or its expression. Thus, LEKTI represents a potential drug for the treatment of these disorders. PMID- 16307659 TI - Plasma cell balanitis treated with tacrolimus 0.1%. AB - BACKGROUND: Plasma cell balanitis or balanitis of Zoon (BZ) is a frequent diagnosis in mature men and lesions are generally localized on the glans but may involve the prepuce; its aetiology remains unknown. OBJECTIVES: To present our experience with tacrolimus 0.1% ointment in the treatment of two patients with BZ refractive to other topical treatments. PATIENTS AND METHODS: Two uncircumcised mature caucasian males were seen, both presenting with BZ; topical tacrolimus 0.1% ointment twice daily was prescribed in each case. RESULTS: Marked improvement of the lesions in both patients has been observed, with follow-up of 1 year and 10 months, respectively. CONCLUSIONS: Topical tacrolimus 0.1% ointment is an effective and safe treatment for BZ. PMID- 16307660 TI - Pathogenetic mechanisms of vitiligo in a patient with Sezary syndrome. AB - Patients exhibiting association between vitiligo and cutaneous T-cell lymphoma (CTCL) remain rare and it is not known whether some T-cell subpopulations of CTCL in the skin are able to recognize specific melanocytic epitopes and thus induce vitiligo. The aim of our study was to determine whether T cells specific to melanocyte differentiation antigens were detectable among tumour-infiltrating lymphocytes (TIL) in the hypopigmented skin of a patient with Sezary syndrome (SS). A 71-year-old patient presented with SS and developed vitiligo during the course of her disease. Immunohistochemical studies showed staining with HMB45 and MelanA antibodies in the pigmented skin biopsy, whereas no staining was observed in the hypopigmented skin biopsy. To analyse responses to melanocyte differentiation antigens, we used a transient COS transfection assay that permits an estimation of CD8 T-cell responses against a large number of HLA/antigen combinations. This technique allowed the detection of melanocyte differentiation antigen-specific T lymphocytes, directed mainly against Melan-A/MART1 antigen in the HLA-A*23 context. Our study supports the concept that vitiligo that has developed during the evolution of a CTCL is related to the presence of a T lymphocyte subpopulation reactive against melanocyte differentiation antigens (mainly Melan-A/MART1) present in skin lesions. The role of interferon in the induction of this T-lymphocyte subpopulation is discussed. PMID- 16307661 TI - Two novel CYLD gene mutations in Chinese families with trichoepithelioma and a literature review of 16 families with trichoepithelioma reported in China. PMID- 16307662 TI - A non-sense mutation in the corneodesmosin gene in a Mexican family with hypotrichosis simplex of the scalp. AB - BACKGROUND: Hypotrichosis simplex of the scalp (HSS; MIM 146520) is a rare autosomal dominant form of non-syndromic alopecia that affects men and women equally. Up to now, only a small number of families with HSS have been reported. The affected individuals experience a diffuse progressing hair loss from childhood to adulthood that is confined to the scalp. Recently, HSS has been mapped to the short arm of chromosome 6 (6p21.3), allowing mutations in the corneodesmosin gene (CDSN) to be identified as the cause of the disorder. To date, two stop mutations have been found in three unrelated families with HSS of different ethnic origin. OBJECTIVES: To describe the first HSS-family with Latin American (Mexican) background comprising 6 generations and to identify a mutation in the CDSN gene. PATIENTS/METHODS: The patients were examined by a clinician and blood samples were taken. After DNA extraction, sequencing analysis of the CDSN gene and restriction enzyme analysis with PsuI were performed. RESULTS: By direct sequencing of the two exons of the CDSN gene, a nonsense mutation was identified in the index patient in exon 2, resulting in a premature stop codon (Y239X). The mutation co-segregates perfectly in the family with the disease and was not found in 300 control chromosomes using a restriction enzyme analysis with PsuI. CONCLUSIONS: A nonsense mutation was identified in the first family with HSS of Latin American ethnical background. Our data provide molecular genetic evidence for a 3rd stop mutation in exon 2 of the CDSN gene being responsible for HSS. All to date known nonsense mutations responsible 3 for HSS are clustered in a region of 40 amino acids which is in accordance with a dominant negative effect conferred by aggregates of truncated CDSN proteins. PMID- 16307663 TI - HHV8-positive Kaposi sarcoma in a long-term human immunodeficiency virus-exposed but uninfected patient carrying the CCR5-promoter mutation A59029G. PMID- 16307664 TI - Severe cutaneous toxicity following treatment with gefitinib (ZD1839). PMID- 16307665 TI - Local hypertrichosis associated with a human papillomavirus type 1-induced wart. PMID- 16307666 TI - Cosmetic dermatology: almost an oxymoron! PMID- 16307667 TI - Development of follicular rash in measles. PMID- 16307668 TI - Vulval syringoma successfully treated with tranilast. PMID- 16307669 TI - Immunohistochemical analyses of p63 expression in cutaneous tumours. PMID- 16307670 TI - Narrow-band ultraviolet B in aquagenic pruritus. PMID- 16307671 TI - Diagnostic phototesting in polymorphous light eruption: the optimal number of irradiations. PMID- 16307672 TI - Pemphigus vegetans with IgG and IgA antidesmoglein 3 antibodies. PMID- 16307673 TI - Myotoxicity occurring with ciclosporin in a patient with atopic dermatitis. PMID- 16307674 TI - Successful use of etretinate for long-term management of a patient with cutaneous type adult T-cell leukaemia/lymphoma. PMID- 16307675 TI - Erythema annulare centrifugum following herpes zoster infection: Wolf's isotopic response? PMID- 16307676 TI - Infliximab can precipitate as well as worsen palmoplantar pustulosis: possible linkage to the expression of tumour necrosis factor-alpha in the normal palmar eccrine sweat duct? PMID- 16307677 TI - Once daily vs. twice daily applications of topical treatments in psoriasis. PMID- 16307678 TI - Annual Congress of the British Society for Immunology, 6-9 December 2005, Harrogate, UK. Abstracts. PMID- 16307679 TI - What factors explain the number of physical therapy treatment sessions in patients referred with low back pain; a multilevel analysis. AB - BACKGROUND: It is well-known that the number of physical therapy treatment sessions varies over treatment episodes. Information is lacking, however, on the source and explanation of the variation. The purposes of the current study are: 1) to determine how the variance in the number of physical therapy treatment sessions in patients with non-specific low back pain (LBP) in the Netherlands is distributed over patient level, therapist level and practice level; and 2) to determine the factors that explain the variance. METHODS: Data were used from a national registration network on physical therapy. Our database contained information on 1,733 patients referred with LBP, treated by 97 therapists working in 41 practices. The variation in the number of treatment sessions was investigated by means of multilevel regression analyses. RESULTS: Eighty-eight per cent of the variation in the number of treatment sessions for patients with LBP is located at patient level and seven per cent is located at practice level. It was possible to explain thirteen per cent of all variance. The duration of the complaint, prior therapy, and the patients' age and gender in particular are related to the number of physical therapy treatment sessions. CONCLUSION: Our results suggest that the number of physical therapy treatment sessions in patients with LBP mainly depends on patient characteristics. More variation needs to be explained, however, to improve the transparency of care. Future research should examine the contribution of psychosocial factors, baseline disability, and the ability to learn motor behavior as possible factors in the variation in treatment sessions. PMID- 16307680 TI - Genetic, household and spatial clustering of leprosy on an island in Indonesia: a population-based study. AB - BACKGROUND: It is generally accepted that genetic factors play a role in susceptibility to both leprosy per se and leprosy type, but only few studies have tempted to quantify this. Estimating the contribution of genetic factors to clustering of leprosy within families is difficult since these persons often share the same environment. The first aim of this study was to test which correlation structure (genetic, household or spatial) gives the best explanation for the distribution of leprosy patients and seropositive persons and second to quantify the role of genetic factors in the occurrence of leprosy and seropositivity. METHODS: The three correlation structures were proposed for population data (n = 560), collected on a geographically isolated island highly endemic for leprosy, to explain the distribution of leprosy per se, leprosy type and persons harbouring Mycobacterium leprae-specific antibodies. Heritability estimates and risk ratios for siblings were calculated to quantify the genetic effect. Leprosy was clinically diagnosed and specific anti-M. leprae antibodies were measured using ELISA. RESULTS: For leprosy per se in the total population the genetic correlation structure fitted best. In the population with relative stable household status (persons under 21 years and above 39 years) all structures were significant. For multibacillary leprosy (MB) genetic factors seemed more important than for paucibacillary leprosy. Seropositivity could be explained best by the spatial model, but the genetic model was also significant. Heritability was 57% for leprosy per se and 31% for seropositivity. CONCLUSION: Genetic factors seem to play an important role in the clustering of patients with a more advanced form of leprosy, and they could explain more than half of the total phenotypic variance. PMID- 16307681 TI - Immunosenescence and vaccination. AB - The problems associated with the ageing immune system and vaccination were discussed recently at an international workshop at the Jenner Institute for Vaccine Research, Compton, UK, 6-7 October, 2005. This is a commentary on that session. The meeting included discussions on T and B cell differentiation and ageing, as well as dendritic cell and neutrophil data, with the emphasis on T cell immunosenescence, perceived as the most important hindrance to satisfactory responses to vaccines in the elderly. The main questions to be addressed in this context are the reasons for dysfunctionality of T cells in the elderly and what to do to improve T cell function. Several of the major reasons for poor T cell responses in the elderly were discussed; however, many important questions remain: The next meeting at the Jenner Institute may already be able to provide some of the answers to these questions, which have serious implications for public health issues in increasingly elderly populations. PMID- 16307682 TI - Failure of levofloxacin treatment in community-acquired pneumococcal pneumonia. AB - BACKGROUND: Streptococcus pneumoniae is the leading cause of community-acquired pneumonia (CAP). High global incidence of macrolide and penicillin resistance has been reported, whereas fluoroquinolone resistance is uncommon. Current guidelines for suspected CAP in patients with co-morbidity factors and recent antibiotic therapy recommend initial empiric therapy using one fluoroquinolone or one macrolide associated to other drugs (amoxicillin, amoxicillin/clavulanate, broad spectrum cephalosporins). Resistance to fluoroquinolones is determined by efflux mechanisms and/or mutations in the parC and parE genes coding for topoisomerase IV and/or gyrA and gyrB genes coding for DNA gyrase. No clinical cases due to fluoroquinolone-resistant S. pneumoniae strains have been yet reported from Italy. CASE PRESENTATION: A 72-year-old patient with long history of chronic obstructive pulmonary disease and multiple fluoroquinolone treatments for recurrent lower respiratory tract infections developed fever, increased sputum production, and dyspnea. He was treated with oral levofloxacin (500 mg bid). Three days later, because of acute respiratory insufficiency, the patient was hospitalized. Levofloxacin treatment was supplemented with piperacillin/tazobactam. Microbiological tests detected a S. pneumoniae strain intermediate to penicillin (MIC, 1 mg/L) and resistant to macrolides (MIC >256 mg/L) and fluoroquinolones (MIC >32 mg/L). Point mutations were detected in gyrA (Ser81-Phe), parE (Ile460-Val), and parC gene (Ser79-Phe; Lys137-Asn). Complete clinical response followed treatment with piperacillin/tazobactam. CONCLUSION: This is the first Italian case of community-acquired pneumonia due to a fluoroquinolone-resistant S. pneumoniae isolate where treatment failure of levofloxacin was documented. Molecular analysis showed a group of mutations that have not yet been reported from Italy and has been detected only twice in Europe. Treatment with piperacillin/tazobactam appears an effective means to inhibit fluoroquinolone-resistant strains of S. pneumoniae causing community-acquired pneumonia in seriously ill patients. PMID- 16307683 TI - Phosphodiesterase 3 inhibition and cough in elderly asthmatics. AB - AIMS: Cough is a common symptom of bronchial asthma, a chronic inflammatory airway disease. Recently, the therapeutic effects of selective phosphodiesterase (PDE) inhibitors have been focused on bronchial asthma. This study was designed to investigate the clinical effect of PDE 3 inhibition on cough reflex sensitivity in elderly patients with bronchial asthma. METHODS: Effects of cilostazol, a PDE 3 inhibitor, on cough response to inhaled capsaicin were examined in 11 patients over 70 years with stable asthma in a randomized, placebo controlled cross over study. Capsaicin cough threshold, defined as the lowest concentration of capsaicin eliciting five or more coughs, was measured as an index of airway cough reflex sensitivity. RESULTS: The cough threshold was significantly (p < 0.05) increased after two-week treatment with cilostazol (100 mg twice a day orally) compared with placebo [48.8 (GSEM 1.4) vs. 29.2 (GSEM 1.3) muM]. CONCLUSION: These findings indicate that PDE 3 inhibition may be a novel therapeutic option for elderly patients with asthma, especially for their cough symptoms. PMID- 16307684 TI - Copper chaperone for Cu/Zn superoxide dismutase is a sensitive biomarker of mild copper deficiency induced by moderately high intakes of zinc. AB - BACKGROUND: Small increases in zinc (Zn) consumption above recommended amounts have been shown to reduce copper (Cu) status in experimental animals and humans. Recently, we have reported that copper chaperone for Cu/Zn superoxide dismutase (CCS) protein level is increased in tissues of overtly Cu-deficient rats and proposed CCS as a novel biomarker of Cu status. METHODS: Weanling male Wistar rats were fed one of four diets normal in Cu and containing normal (30 mg Zn/kg diet) or moderately high (60, 120 or 240 mg Zn/kg diet) amounts of Zn for 5 weeks. To begin to examine the clinical relevance of CCS, we compared the sensitivity of CCS to mild Cu deficiency, induced by moderately high intakes of Zn, with conventional indices of Cu status. RESULTS: Liver and erythrocyte CCS expression was significantly (P < 0.05) increased in rats fed the Zn-60 and/or Zn 120 diet compared to rats fed normal levels of Zn (Zn-30). Erythrocyte CCS expression was the most sensitive measure of reduced Cu status and was able to detect a decrease in Cu nutriture in rats fed only twice the recommended amount of Zn. Liver, erythrocyte and white blood cell CCS expression showed a significant (P < 0.05) inverse correlation with plasma and liver Cu concentrations and caeruloplasmin activity. Unexpectedly, rats fed the highest level of Zn (Zn-240) showed overall better Cu status than rats fed a lower level of elevated Zn (Zn-120). Improved Cu status in these rats correlated with increased duodenal mRNA expression of several Zn-trafficking proteins (i.e. MT-1, ZnT-1, ZnT-2 and ZnT-4). CONCLUSION: Collectively, these data show that CCS is a sensitive measure of Zn-induced mild Cu deficiency and demonstrate a dose dependent biphasic response for reduced Cu status by moderately high intakes of Zn. PMID- 16307685 TI - Patient and health service delay in pulmonary tuberculosis patients attending a referral hospital: a cross-sectional study. AB - BACKGROUND: Delays in diagnosis and initiation of effective treatment increase morbidity and mortality from tuberculosis as well as the risk of transmission in the community. The aim of this study was to determine the time taken for patients later confirmed as having TB to present with symptoms to the first health provider (patient delay) and the time taken between the first health care visit and initiation of tuberculosis treatment (health service delay). Factors relating to these 'delays' were analyzed. METHODS: A cross-sectional survey, of 231 newly diagnosed smear-positive tuberculosis patients was conducted in Mulago National referral Hospital Kampala, from January to May 2002. Socio-demographic, lifestyle and health seeking factors were evaluated for their association with patient delay (> 2 weeks) and health service delay (> 4 weeks), using odds ratios with 95% confidence intervals (CI) including multivariate logistic regression. RESULTS: The median total delay to treatment initiation was 12 weeks. Patients often presented to drug shops or pharmacies (39.4%) and private clinics (36.8%) more commonly than government health units (14%) as initial contacts. Several independent predictors of 'patient delay' were identified: being hospitalized (odds ratio [0R] = 0.32; 95% CI: 0.12-0.80), daily alcohol consumption (OR = 3.7; CI: 1.57-9.76), subsistence farming (OR = 4.70; CI: 1.67-13.22), and perception of smoking as a cause of TB (OR = 5.54; CI: 2.26-13.58). Independent predictors of 'health service delay' were: > 2 health seeking encounters per month (OR = 2.74; CI: 1.10-6.83), and medical expenditure on TB related symptoms > 29 US dollars (OR = 3.88; CI: 1.19-12.62). Perceived TB stigma and education status was not associated with either form of delay. CONCLUSION: Delay in diagnosis of TB is prolonged at the referral centre with a significant proportion of Health service delay. More specific and effective health education of the general public on tuberculosis and seeking of appropriate medical consultation is likely to improve case detection. Certain specific groups require further attention. Alcoholics and subsistence farmers should be targeted to improve accessibility to TB treatment. Continuing medical education about TB management procedures for health providers and improvement in the capacity of TB control services should be undertaken. PMID- 16307686 TI - Detection of human papillomavirus DNA and p53 codon 72 polymorphism in prostate carcinomas of patients from Argentina. AB - BACKGROUND: Infections with high-risk human papillomaviruses (HPVs), causatively linked to cervical cancer, might also play a role in the development of prostate cancer. Furthermore, the polymorphism at codon 72 (encoding either arginine or proline) of the p53 tumor-suppressor gene is discussed as a possible determinant for cancer risk. The HPV E6 oncoprotein induces degradation of the p53 protein. The aim of this study was to analyse prostate carcinomas and hyperplasias of patients from Argentina for the presence of HPV DNA and the p53 codon 72 polymorphism genotype. METHODS: HPV DNA detection and typing were done by consensus L1 and type-specific PCR assays, respectively, and Southern blot hybridizations. Genotyping of p53 codon 72 polymorphism was performed both by allele specific primer PCRs and PCR-RFLP (Bsh1236I). Fischer's test with Woolf's approximation was used for statistical analysis. RESULTS: HPV DNA was detected in 17 out of 41 (41.5%) carcinoma samples, whereas all 30 hyperplasia samples were HPV-negative. Differences in p53 codon 72 allelic frequencies were not observed, neither between carcinomas and hyperplasias nor between HPV-positive and HPV negative carcinomas. CONCLUSION: These results indicate that the p53 genotype is probably not a risk factor for prostate cancer, and that HPV infections could be associated with at least a subset of prostate carcinomas. PMID- 16307687 TI - A Chiropracticness Test. AB - BACKGROUND: There is little homogeneity of opinion in the chiropractic profession about its essence and identity. Matters compromising the establishment of a coherent identity include the issue of vertebral subluxation, philosophy, mercantilism, poverty of qualifications in some chiropractic college faculty, and lack of intellectual productivity in some chiropractic college faculty. DISCUSSION: The Chiropractic profession has mislabeled rhetoric, supposition and cant as philosophy, whilst showing sparse evidence for the existence of more than a few chiropractors writing in philosophy as a discipline. There is no evidence for "Chiropractic Philosophy". I propose, however, that a better use of the discipline of philosophy can be of great use to the Chiropractic profession. Various thinkers throughout the ages have written about deduction, induction and falsificationism as methods to discover more reliably the nature of things in the world about us. Each method has strengths and frailties, but some of the latter are insurmountable for our purposes. SUMMARY: Using a contrivance of that method which seems most suited, sui generis, for the purpose, I propose a Chiropracticness Test as a tool to assist the search for essence and identity in Chiropractic. PMID- 16307688 TI - Catheter-associated bacteremia by Mycobacterium senegalense in Korea. AB - BACKGROUND: Rapidly growing mycobacteria is recognized as one of the causative agents of catheter-related infections, especially in immunocompromised hosts. To date, however, Mycobacterium senegalense, which was known as the principal pathogen of bovine farcy, has not been reported in human infection. CASE PRESENTATION: We describe the first case of human infection by M. senegalense, which has caused catheter-related bloodstream infection in a cancer patient in Korea. The microorganism was identified by the 16S rRNA gene, rpoB, and 16S-23S rRNA gene internal transcribed spacer (ITS) sequence analyses. CONCLUSION: Our first report of catheter-associated bacteremia caused by M. senegalense suggests the zoonotic nature of this species and indicates the expansion of mycobacterial species relating to human infection. M. senegalense should be considered as one of the causes of human infections in the clinical practice. PMID- 16307689 TI - The efficacy of tadalafil in improving sexual satisfaction and overall satisfaction in men with mild, moderate, and severe erectile dysfunction: a retrospective pooled analysis of data from randomized, placebo-controlled clinical trials. AB - BACKGROUND: Satisfaction with the sexual experience is considered important when evaluating the impact of treatments for erectile dysfunction (ED), yet satisfaction has been infrequently assessed in clinical trials. OBJECTIVE: To evaluate satisfaction with, and enjoyment of, the sexual experience in men with ED enrolled in 11 placebo-controlled clinical trials of tadalafil. STUDY DESIGN AND METHODS: Retrospective pooled analysis of data from 11 randomized, double blind, placebo-controlled clinical trials of tadalafil. Men with mild (N = 838), moderate (N = 558), or severe (N = 703) ED who were randomized to tadalafil 10 mg or 20 mg or placebo taken as needed for 12 weeks were included in this analysis. Efficacy measures included the International Index of Erectile Function (IIEF). Reported herein are the scores on the IIEF overall satisfaction domain and individual IIEF questions (IIEF-Q7, satisfaction with intercourse; and IIEF-Q8, enjoyment of intercourse). RESULTS: At least moderate satisfaction (IIEF overall satisfaction domain) was reported by 55% and 72% of patients with mild ED taking tadalafil 10 mg and 20 mg, respectively, compared with 33% taking placebo (p < 0.002); 60% and 65% vs. 19% of patients with moderate ED (p < 0.001); and 32% and 49% vs. 9% with severe ED (p < 0.001). Satisfactory intercourse during most attempts or almost always/always (IIEF-Q7) was reported by 59% and 79% of patients with mild ED taking tadalafil 10 mg and 20 mg vs. 32% taking placebo (p < 0.001); 52% and 65% vs. 18% with moderate ED (p < 0.001); and 28% and 49% vs. 5% with severe ED (p < 0.001). Highly or very highly enjoyable intercourse (IIEF Q8) was reported by 45% and 63% of patients with mild ED taking tadalafil 10 mg and 20 mg vs. 21% taking placebo (p < 0.001); 43% and 56% vs. 16% with moderate ED (p < 0.001); and 19% and 44% vs. 5% with severe ED (p < 0.001). CONCLUSIONS: Compared with placebo, tadalafil 10 mg and 20 mg improved overall satisfaction with the sexual experience, intercourse satisfaction, and intercourse enjoyment in men with mild, moderate, and severe ED. PMID- 16307690 TI - Efficacy of Rhizophora mangle aqueous bark extract (RMABE) in the treatment of aphthous ulcers: a pilot study. AB - OBJECTIVE: Rhizophora mangle aqueous bark extract (RMABE) (CIKRON-H), has been used as antiseptic and skin wound healing promoter. The present study was a randomised, single-blinded, placebo control trial conducted to asses the efficacy of RMABE in treating oral aphthous ulcers. RESEARCH DESIGN AND METHODS: Patients (n = 32) with aphthous ulcers were randomised to received placebo solution or RMABE topically, once a day, from Monday to Friday, until they healed. The efficacy of the treatment was evaluated by physician clinical observations (time to heal, change in condition), the quality of the patient's life and the tolerability through recording adverse effects. RESULTS: No demographic differences were noted between the two groups at base-line. Seven days after treatment, 12 of the 17 patients in the RMABE group (71%) were completely healed of their aphthous ulcers, with repaired mucosa and no symptoms of ulcers, compared with one in 15 patients in the placebo group (7%) (p < 0.0001). The time taken for the signs and symptoms of ulcers to diminish was also higher in the placebo than in RMABE-treatment group (erythema: placebo 10.54 +/- 1.24, RMABE 4.94 +/- 0.72 days, p = 0.0003; ardour: placebo 7.00 +/- 0.76, RMABE 2.93 +/- 0.49 days, p = 0.0001; and pain: placebo 7.43 +/- 1.21, RMABE 2.92 +/- 0.23 days, p = 0.0011). No subject showed any sign of adverse effects. CONCLUSIONS: These observations demonstrate that the R. mangle aqueous bark extract reduced the time to repair mucosal tissue, erythema, ardour and pain persistence. There was no evidence any adverse effects. This is the first time that the R. mangle extract has been reported to have mouth mucosa healing properties. PMID- 16307691 TI - Dyslipidaemia and cardiovascular risk in HIV infection. AB - Cardiovascular disease is increasingly recognised as a consequence of human immunodeficiency virus (HIV) infection in the era of highly active antiretroviral therapy (HAART). Dyslipidaemia is also a feature of HIV infection itself and the antiretroviral drugs from the protease inhibitor classes. Increased rates of atherosclerotic disease and diabetes have been associated with lipodystrophy and now from one of the major causes of morbidity in HIV and acquired immunodeficiency syndrome (AIDS).This review, based on a multi-database keyword search, summarises the lipid changes observed in the course of HIV infection and its treatment, and puts them into the context of other risk factors for cardiac disease, and other causes of cardiovascular disease in HIV. PMID- 16307692 TI - Overall tolerability and analgesic activity of intra-articular sodium hyaluronate in the treatment of knee osteoarthritis. AB - OBJECTIVE: Viscosupplementation with intra-articular hyaluronic acid (HA) is an alternative to the treatment of symptomatic knee osteoarthritis (OA) with pain relieving drugs. Sinovial, is a sterile, non-pyrogenic 0.8% solution of highly purified sodium hyaluronate for intra-articular application. The aim of the present study was to investigate the safety and tolerability profile of this preparation in patients with symptomatic knee OA over 24 weeks. RESEARCH DESIGN AND METHODS: This was a single group, open-label study, including outpatients of both sexes, aged between 18 and 85 years, with symptomatic knee OA. All patients underwent weekly intra-articular injections of HA for 5 consecutive weeks and were followed-up for 19 additional weeks. The safety and tolerability profile (primary endpoint) was assessed by adverse event (AE) reporting. The secondary endpoint was efficacy evaluated by changes in the Western Ontario and McMaster Universities (WOMAC) score vs. baseline. Patient and physician satisfaction were also recorded. RESULTS: Intra-articular HA was generally well tolerated. The most frequent AE was pain at the injection site (5.8% of the injections); no serious treatment-related AE was reported. The WOMAC score was significantly reduced within the first 2 weeks of treatment (from 4.02 +/- 1.90 to 3.55 +/- 2.04, p = 0.0011), further decreased by the end of the injection series (week 6: 2.59 +/- 1.90; p < 0.0001) and maintained during the follow-up (week 24: 2.44 +/- 1.88; p < 0.0001). The WOMAC subscores were also significantly reduced from week 4 for 'pain' and from week 6 for 'stiffness' and 'physical function'. CONCLUSIONS: In the present study, intra-articular HA was well tolerated and safe in patients with symptomatic knee OA. Based on the sustained improvements in WOMAC score and subscores, a carry-over effect lasting for at least 19 weeks after the last injection may be proposed. These results further confirm the evidence of efficacy and safety of intra-articular HA in the management of knee OA. PMID- 16307693 TI - Treating dry skin and pruritus with a bath oil containing soya oil and lauromacrogols. AB - The efficacy and safety of a bath oil containing soya oil and anti-pruritic lauromacrogols have been demonstrated in experimental and clinical studies. In order to generate more efficacy and safety-related data on the day-to-day use in paediatric practice, a post-marketing surveillance study has been conducted. More than 3500 patients--the vast majority (61.4%) aged 0-4 years--were included in this investigation. Atopic eczema was diagnosed in 86.4% of the cases. The average duration of treatment was 42 days. The results confirm that the anti pruritic bath oil is both well-tolerated and effective in treating dry and itchy skin diseases. The sum score of symptoms decreased during the treatment period from 7.21 to 2.71 mean score points. The physicians assessed the global efficacy in 89.4% of the cases as 'very good' or 'good'. Anti-pruritic properties of the combination were rated 'very good' or 'good' in 84.3%. The local tolerability was stated to be 'good' or 'very good' in 96.8% of the patients. In total, only 10 out of the 3566 patients, 0.28% respectively, experienced discomfort.Moreover, the physicians estimated that the bath oil therapy helped to reduce therapeutics like steroids in nearly 60% of patients. The potential to reduce use of external steroids and other specific therapeutics is also of particular relevance from a pharmacoeconomic point of view. PMID- 16307694 TI - Endoscopic and histologic evidence of persistent mucosal healing and correlation with clinical improvement following sustained infliximab treatment for Crohn's disease. AB - OBJECTIVES: The long-term effect of infliximab on endoscopic and histologic disease activity and expression of inflammatory markers was assessed in Crohn's disease patients who received infliximab as episodic or scheduled maintenance therapy therapy over 54 weeks (ACCENT 1). METHODS: All patients received Infliximab 5 mg/kg at week 0 and at week 2 were then randomized as responders or nonresponders to placebo or infliximab (5 or 10mg/kg). Patients received placebo or infliximab 5 mg/kg at weeks 2 and 6 followed by placebo or infliximab (5 or 10mg/kg) every 8 weeks or episodically on loss of response. Crohn's Disease Activity Index (CDAI), Crohn's Disease Endoscopic Index of Severity (CDEIS), Inflammatory Bowel Disease Questionnaire (IBDQ), and colonic and ileal Global Histologic Disease Activity (CGHAS and IGHAS) scores were determined at weeks 0, 10, and 54. Tumor necrosis factor-alpha (TNF-alpha), gelatinase B, Infliximab, tenascin, clusters of differentiation marker 68 (CD68), and intercellular adhesion molecule-1 (ICAM-1) were detected in mucosal biopsies by immunohistochemistry. RESULTS: At baseline, CDEIS significantly correlated with CGHAS only. Further at baseline, both CDEIS and the worst CGHAS or IGHAS, were significantly correlated with CD68, ICAM-1, and gelatinase B expression. At week 10, improvement in CGHAS only, correlated significantly with better CDAI, CDEIS, and IBDQ scores. Improvements in CDEIS and GHAS at week 10 correlated with reductions in gelatinase B and CD68, whereas only GHAS improvement correlated with decreased TNF-alpha expression. At week 54, decreased gelatinase B expression continued to correlate with improved CDEIS and GHAS while decreased CD68 and TNF-alpha expression correlated with GHAS and CDEIS improvement, respectively. CONCLUSIONS: Endoscopic and histologic evidence of mucosal healing was associated with a sustained reduction in the expression of inflammatory markers. Infliximab-induced improvement in the clinical signs and symptoms of Crohn's disease was associated with endoscopic and histologic evidence of sustained mucosal healing. PMID- 16307695 TI - Relationship between obesity and cardiovascular risk factors: findings from a multi-state screening project in the United States. AB - OBJECTIVE: This study examined the impact of body mass index (BMI) category on cardiovascular risk factors such as systolic blood pressure (SBP), high-density lipoprotein (HDL) and total cholesterol (TC). METHODS: Voluntary coronary heart disease (CHD) risk screenings were conducted in 18 states. A mobile screenings unit and team were used to collect data. Respondents were classified as per World Health Organization (WHO) guidelines based on BMI as non-overweight (BMI < or = 25); overweight (BMI > 25 and < or = 30); obese (BMI > 30 and < or = 35); and severely obese (BMI > 35). Hierarchical multiple linear regression analyses were used to measure the impact of BMI on SBP, HDL, and TC after adjusting for age, race, gender, smoking, self-reported prevalence of hypertension and hyperlipidemia, presence of diabetes/CHD event. RESULTS: Of the 12573 screened, 36% were overweight, 16.9% were obese and 7.9% were severely obese. Diabetes and prior CHD event were reported in 5.8% and 12.5% of the respondents, respectively. Mean 10-years CHD risk scores were significantly higher in males (10.4 +/- 9.3) than females (3.2 +/- 4.8) (p < 0.001). Compared to non-overweight respondents, SBP increased by 13.2 mmHg for severely obese (p < 0.001); by 8.9 mmHg for obese (p < 0.001), and by 5.2 mmHg (p < 0.001) for overweight respondents, respectively. TC was 6.8 mg/dL higher in obese (p < 0.01) and 6.9 mg/dL higher in overweight respondents (p < 0.001) as compared to non-overweight respondents. As compared to non-obese respondents, HDL was 9.8 mg/dL lower in severely obese (p < 0.001), 7.6 mg/dL lower in obese (p < 0.001), and 4.6 mg/dL lower in overweight respondents (p < 0.001). CONCLUSIONS: Our results demonstrate a significant impact of increasing BMI category on the CHD risk factors of SBP and HDL in a Us population. These data illustrate the importance of weight reduction in cardiovascular health. PMID- 16307696 TI - Comparison of the efficacy and safety of combinations of metopimazine or ondansetron with methylprednisolone in the prevention of delayed emesis in patients receiving chemotherapy. AB - BACKGROUND: Delayed emesis following chemotherapy in cancer patients remains an important challenge for treatment and contributes to poor quality of life and treatment compliance. OBJECTIVES: To compare the efficacy and tolerability of associations of metopimazine and ondansetron with methylprednisolone for the prevention of delayed chemotherapy-induced nausea and emesis. METHODS: A randomised, open-label, observational, cross-over design was used to compare two treatment strategies following two consecutive sessions of chemotherapy separated by at least 1 week. Patients were randomised to treatment with sublingual metopimazine (15 mg tid) or ondansetron lyophilisate (8 mg bid) for 5 days. All patients received oral methylprednisolone (48 mg). Patients reported episodes of nausea and emesis in a diary, and completed the Functional Living Index Emesis quality of life questionnaire. Adverse events were also evaluated. RESULTS: Ninety-nine patients were included in the study, 79.5% of whom were women, with a mean age of 52.7 years. Breast cancer was the most common individual cancer and most patients were receiving combinations of cytotoxic drugs. Treatment was successful at preventing delayed emesis in 73.6% of patients during treatment with the metopimazine-methylprednisolone association and 57.5% during the ondansetron-methylprednisolone association. Analysis of discordant pairs revealed a significant benefit in favour of the methopimazine-methylprednisolone association (p = 0.006). No significant difference was observed between treatments for the overall quality of life score. The incidence of gastrointestinal disorders, particularly constipation, was significantly higher during ondansetron-methylprednisolone treatment (p = 0.0112). CONCLUSION: Methopimazine is an effective and well-tolerated alternative to setrons for the treatment of delayed nausea and emesis in patients undergoing chemotherapy. PMID- 16307697 TI - Australian attitudes to DNA sample banks and genetic screening. AB - An exploration via an anonymous questionnaire of Australian public attitudes towards medical genetics and sample banking revealed the overwhelming majority views these developments with thoughtful confidence. Continued public education and awareness of these issues will allow the public to make informed decisions and enhance vigilance towards the sometimes misleading coverage in the press and media. PMID- 16307698 TI - Characterisation and comparison of health-related quality of life for patients with renal failure. AB - OBJECTIVE: The objective of this study was to assess the health related quality of life (HRQOL) in patients with kidney failure who had received renal transplants compared to those receiving haemodialysis, peritoneal dialysis or were waiting to start dialysis. RESEARCH DESIGN AND METHODS: The study was conducted at the University Hospital of Wales, Cardiff. HRQOL was measured using the EQ-5D, SF-36 and the Kidney Disease Quality of life questionnaire (KDQOL). Patients with kidney failure were identified from the renal unit departmental database and were surveyed by postal questionnaire or during their treatment. RESULTS: Of 1251 people surveyed, 416 valid returns were received, a response rate of 33%. For renal transplant patients the mean EQ-5Dindex was 0.712 (SD 0.272), significantly higher than those in the other treatment groups (haemodialysis mean = 0.443 (SD 317), p < 0.001; peritoneal dialysis mean = 0.569 (SD 329), p < 0.001). This difference remained after controlling for age and co morbidity. With the exception of pain, the SF-36 showed significantly higher scores across all domains for transplant patients compared to both dialysis groups. From the KDQOL there were significantly lower scores compared with the transplant patients for both groups of dialysis patients for the effects and burden of kidney disease and general symptoms and problems. However, overall health scores were significantly higher for dialysis patients compared with transplant patients. CONCLUSION: Kidney failure has a high cost in terms of health related quality of life. There was a large difference between patients who have received a functioning graft following kidney transplant versus the alternative methods of renal replacement therapy, that is, peritoneal dialysis and haemodialysis. Kidney transplant should be the treatment of choice, and every effort should be made to increase the availability of kidneys for transplantation. PMID- 16307699 TI - An international survey of insomnia: under-recognition and under-treatment of a polysymptomatic condition. AB - BACKGROUND: Due to its high prevalence, considerable impact on well-being, and high medical and societal costs, insomnia represents an important healthcare challenge. Despite this, the prevalence of this condition is under-recognized and many sufferers do not receive adequate treatment. OBJECTIVE: This international survey investigated the prevalence and characteristics of insomnia in the general population in France. Italy, Japan and the USA to better understand why insomnia is under-recognized and under-treated. RESEARCH DESIGN AND METHODS: Type, frequency, natural history of night- and day-time symptoms, and attitudes of people regarding the nature and treatment of insomnia was quantitatively assessed from a representative sample of the general population aged > 18 years in each of the four countries. A survey of an extensive series of standardized 10-15-min interviews was conducted over the telephone by professional interviewers. RESULTS: Insomnia was reported by 37.2% of respondents in France and Italy, 6.6% in Japan, and 27.1% in the USA. The mean number of symptoms reported per patient was two, with sleep maintenance insomnia the most predominant symptom (73%), followed by difficulty falling asleep (61%), and poor sleep quality (48%). Daytime fatigue and impaired concentration and attention were the most commonly reported next-day symptoms as a result of poor sleep. The majority of individuals with sleep problems reported being 'somewhat' or 'very' bothered by their symptoms, and reported that their sleep problems impacted on their daily quality of life either 'somewhat' or 'a lot'. Among individuals with a history of insomnia, the rate of reporting insomnia symptoms to physicians was generally low and of those who did consult a physician, few were prescribed any medication. CONCLUSIONS: The results of this study suggest that in Europe, Japan and the USA, the burden of insomnia on sufferers is considerable. Many respondents took no action to alleviate their insomnia symptoms despite the poor sleep impacting on their daily quality of life either 'somewhat' or 'a lot'. PMID- 16307700 TI - Patterns of graft and patient survival following renal transplantation and evaluation of serum creatinine as a predictor of survival: a review of data collected from one clinical centre over 34 years. AB - BACKGROUND: The pattern of renal transplantation has never been described since the introduction of the technique. The purpose of this study was therefore to characterise the pattern of renal transplantation from 1967 to 2000, focusing on renal graft function as a predictor of survival. METHODS: This study was a retrospective analysis of an electronic database. The setting was a single renal transplant centre in the United Kingdom covering a population of 2.2 million and included patients who received at least one renal transplant over the study period (n = 1516). The main outcome measures were patient and graft survival, acute rejection episodes and patterns of graft function, as measure by creatinine levels. RESULTS: There were 559 (36.8%) female patients; 109 (7.2%) patients had pre-existing diabetes. Patient survival was adversely affected by increased age at transplant (p < 0.001): 5-year patient survival from first transplant was 82% for patients aged 0 to 17 years, 80% for 18 to 49 years and 61% for > 49 years. Pre-existing diabetes also adversely affected survival (p < 0.01): 5-year graft survival was 63% for patients with diabetes versus 74% for those without. Graft survival was significantly associated with serum creatinine levels recorded 1 year post-primary transplant (p < 0.001) and with three or more acute rejection episodes (p < 0.05). Neither gender nor diabetes status were statistically significant in predicting graft survival. The number of acute rejection episodes was significantly greater in patients with pre-existing diabetes than those without (61% versus 42%, respectively; p < 0.001). There were no differences in the number of acute rejection episodes occurring across age groups. CONCLUSION: Patient and graft survival improved markedly over the 34-year study period, although patient survival has changed little since 1990. Serum creatinine levels are a reliable predictor of graft survival. PMID- 16307701 TI - An examination of self-reported chronic conditions and health status in the 2001 Medicare Health Outcomes Survey. AB - OBJECTIVES: To estimate the prevalence of chronic conditions in managed care enrolled Medicare seniors (age 65 years plus) and to examine the association between self-reported chronic conditions and health status, as measured by the SF 36. METHODS: Data were obtained from the 2001 Medicare Health Outcomes Survey (HOS). The HOS is conducted to assess the quality of care provided to Medicare beneficiaries enrolled in managed care. The survey questionnaire, which was administered by phone or mail, includes the SF-36 and items addressing demographic characteristics, depressive symptoms, and chronic conditions. The SF 36 produces eight multi-item scale scores and physical and mental component summary scores. For this analysis, an ordinary least squares regression model was conducted using the SF-36 scales and summary scores as dependent variables to estimate the association between each chronic condition and the scale scores after adjusting for demographic variables. RESULTS: More than three-fourths of the respondents had at least one chronic condition. Among the conditions, hypertension (56.6%) arthritis of the hip or knee (39.7%) and arthritis of the hand or wrist (33.3%) were the most commonly reported. Compared with other variables, age and arthritis were most highly associated with the SF-36 measures reflective of physical health. Depressive symptoms had the strongest association with the SF-36 measures most reflective of mental health. Among the chronic conditions, the adverse impact of having difficulty in controlling urination, a relatively neglected condition, was only second to depressive symptoms in its negative relationship with vitality, social-functioning, and mental health. CONCLUSION: Chronic conditions were commonly reported among the older adults. The unique associations found between chronic conditions and domains of health status demonstrate the importance of examining the burden of these conditions in terms of functioning and well-being. The findings of this study may help inform decision making at the patient, health plan, and societal levels. PMID- 16307702 TI - Effects of switching from an AChE inhibitor to a dual AChE-BuChE inhibitor in patients with Alzheimer's disease. AB - OBJECTIVE: Cholinesterase (ChE) inhibitors are the only medications approved for the treatment of Alzheimer's disease (AD). The features of ChE inhibitors differ considerably. In addition to acetylcholinesterase (AChE) inhibition, rivastigmine also inhibits butrylcholinesterase (BuChE), providing dual AChE and BuChE inhibition. An observational study was performed to determine the response in routine clinical practice to switching AD patients to rivastigmine from a selective AChE inhibitor when that treatment no longer delivered a satisfactory clinical response. RESEARCH DESIGN AND METHODS: A prospective, multicentre, 3 month observational trial in patients with mild to moderately severe AD (adjusted Mini Mental State Examination [MMSE] score 10-26) deteriorating (at least 2 adjusted MMSE points in last 6 months) on selective AChE inhibitor treatment. Adjusted MMSE, activities of daily living (ADL) and instrumental activities of daily living (IADL), the Zarit caregiver burden and global function (short Clinical Global Impression of Change, CGIC) scores were noted before the switch and 3 months after the switch. RESULTS: 225 patients entered the study. The switches made were from donepezil to rivastigmine in (D-R) in 188 patients, galantamine to rivastigmine (G-R) in 33 patients and donepezil to galantamine (D G) in four patients. Ten patients discontinued due to adverse events and eight for other reasons. More than half of the switches were within 36 hours of a patient's first treatment visit. In the D-R and G-R groups, 67.7% and 66.7% of patients responded (CGIC score < or = 4), respectively. In non-responders, worsening (CGIC score 5-7) was mild in approximately 80% or more of patients. Adjusted MMSE improved after the switch from both donepezil and galantamine to rivastigmine (+0.69 +/- 3.2, p = 0.008 and +0.6 +/- 1.6, p = 0.05, respectively). Mean ADL, IADL, and Zarit scores remained stable. The proportion of patients on concomitant antipsychotic therapy diminished by 30.5% and benzodiazepines were discontinued in all patients, except one. CONCLUSIONS: AD patients deteriorating on selective AChE inhibitor treatment can benefit from switching to a dual AChE BuChE inhibitor, such as rivastigmine, in terms of stabilization of disease, improvement in cognitive function and reduction in the burden of concomitant psychoactive treatment. The switch was well tolerated. Confirmation of these results is required in a controlled study. PMID- 16307703 TI - Do strong opioids have a role in the early management of back pain? Recommendations from a European expert panel. AB - BACKGROUND: Since chronic low back pain (CLBP) is a complex biopsychosocial problem the ideal treatment is multimodal and multidisciplinary. However, in many countries, primary-care physicians care for many people with CLBP and have a pivotal role in selecting patients for more intensive treatments when these are available. Guidelines on the general use of strong opioids in chronic non-cancer pain have been published but, until now, no specific guidelines were available on their use in chronic low back pain. Given the prevalence of CLBP, and the complex nature of this multifactorial condition, it was felt that specific, evidence based recommendations, with a focus on primary-care treatment, would be helpful. METHODS: An expert panel drawn from across Europe including pain specialists, anaesthetists, neurologists, rheumatologists, a general practitioner, an epidemiologist and the chairman of a pain charity was therefore convened. The aim of the group was to develop evidence-based recommendations that could be used as a framework for more specific guidelines to reflect local differences in the availability of specialist pain services and in the legal status and availability of strong opioids. Statements were based on published evidence (identified by a literature search) wherever possible, and supported by clinical experience when suitable evidence was lacking. RECOMMENDATIONS: Strong opioids have a role in the treatment of low back pain when other treatments have failed. They should be prescribed as part of a multimodal, and ideally interdisciplinary, treatment plan. The aim of treatment should be to relieve pain and facilitate rehabilitation. PMID- 16307704 TI - The psychosocial impact of hair loss among men: a multinational European study. AB - OBJECTIVE: Hair plays an important role in determining self-image, social perceptions, and psychosocial functioning. The objectives of this survey were to identify the impact of hair loss on the self-image of men in five European countries and their level of concern about hair loss with regard to image and personal attractiveness. Moreover, we evaluated participants' use of treatments for hair loss and whether treatment success was associated with psychological benefit. RESEARCH DESIGN AND METHODS: A structured questionnaire of closed-ended questions was administered by telephone in major cities in Germany, France, Italy, Spain, and the United Kingdom. Random sampling was used, and eligible respondents were men from 18 to 45 years old. RESULTS: Of the 1536 men responding to the telephone survey, 729 (47%) reported hair loss; these men are the subject of this report. Over 70% of these men reported hair to be an important feature of image, and 62% agreed that hair loss could affect self-esteem. The realization that they were losing hair was linked to concern about losing an important part of personal attractiveness (43% of men), fear of becoming bald (42%), concern about getting older (37%), negative effects on social life (22%), and feelings of depression (21%). Reduced self-confidence in personal attractiveness was also reported by 38% of men who were not in stable romantic relationships. Less than 10% of men were currently pursuing treatment for hair loss, and three out of four had never pursued treatment for hair loss, either at present or in the past. Those few men who pursued treatment and reported success (n = 73) also reported psychosocial benefits as a result: from 43% to 59% experienced improvements in parameters of self-esteem and perception of personal attractiveness. Reliance on self-reporting of hair loss and use of nonvalidated instruments to measure psychological outcomes are important study limitations. CONCLUSIONS: The results of this survey indicate a gap between the need for treatment of hair loss and initiation of such treatment among men in five European countries. Further research is needed into the factors affecting men's willingness to seek treatment for hair loss. PMID- 16307705 TI - A Markov modelled pharmacoeconomic analysis of bimatoprost 0.03% in the treatment of glaucoma as an alternative to filtration surgery in Italy. AB - OBJECTIVE: Glaucoma is generally managed by decreasing the intraocular pressure (IOP) to a level believed to prevent further damage to the optic disc and loss of visual field. This may be achieved medically or surgically. The objective of this pharmacoeconomic analysis was to investigate the 4-year costs of bimatoprost 0.03% (Lumigan) eye drops as an alternative to filtration surgery (FS) for glaucoma patients on maximum tolerable medical therapy (MTMT). RESEARCH DESIGN AND METHOD: A Markov model was designed using effectiveness and resource use data from a randomized clinical trial and expert statements (Delphi panel). The RCT covered 83 patients on MTMT. The Model compared bimatoprost with FS. In the bimatoprost model arm patients began treatment with bimatoprost. If target IOP ( 20%) was not reached using medical therapy the patient proceeded with FS. In the FS model arm, FS was performed after the first ophthalmologist visit. Unit costs were obtained from an Italian chart and tariffs review (healthcare sector perspective). RESULTS: The RCT showed that 74.7% of the patients delayed the need for FS by 3 months. The Markov model forecasted that 64.2% of the patients could delay the need for FS by 1 year, and forecasted 34.0% could avoid FS after 4 years. The 4-year cost per patient in the bimatoprost and FS arms was E3438 and E4194, respectively (incremental costs of E755). The major cost drivers for the bimatoprost arm were patients who needed combination therapy or FS if the target IOP was not reached. In the FS arm, the major cost drives were the initial surgery costs and pressure-lowering medications used as add-on therapy after FS. CONCLUSIONS: The analysis shows that in a 4-year perspective bimatoprost is cheaper compared to FS. In addition, the postponement of FS associated with bimatoprost may have important implications for waiting list planning. PMID- 16307706 TI - A review of entecavir in the treatment of chronic hepatitis B infection. AB - BACKGROUND: Infection with the hepatitis B virus (HBV) affects two billion people worldwide, and an estimated 400 million people are chronically infected. Currently, FDA-approved regimens for the treatment of chronic HBV include interferon-alpha2b, peginterferon-alpha2a, lamivudine, adefovir dipivoxil, and recently, entecavir. OBJECTIVE: The purpose of this review is to evaluate the pharmacokinetic and pharmacodynamic properties, and the clinical efficacy and safety of entecavir in the treatment of nucleoside-naive and nucleoside-resistant HBeAg-positive and HBeAg-negative chronic hepatitis B (CHB). SEARCH METHODOLOGY: Computerized searches of PubMed and International Pharmaceutical Abstracts from 1985 to July 10, 2005, were performed with the search headings: entecavir, BMS 200475, and chronic hepatitis B. FINDINGS: Entecavir, a new deoxyguanosine analog, represents a third agent within the nucleoside/nucleotide HBV polymerase inhibitor class with distinct advantages over lamivudine and adefovir dipivoxil: it has a three-step mechanism of action, is the most potent inhibitor of HBV DNA polymerase, is not associated with any major adverse effects, and has a limited potential for resistance. In phase II and III clinical trials, entecavir was found to be superior to lamivudine for all primary endpoints evaluated in both nucleoside-naive and lamivudine-resistant patients. Entecavir was effective in both HBeAg-positive and HBeAg-negative nucleoside-naive patients. At this time, optimal duration of entecavir therapy is unknown. CONCLUSION: Entecavir represents a new first- or second-line treatment option for patients chronically infected with HBV. Long-term efficacy and safety studies as well as studies of entecavir in combination with interferon products or other nucleoside/nucleotide analogs are eagerly awaited. PMID- 16307707 TI - Effect of nesiritide on renal function: a retrospective review. AB - OBJECTIVES: Recent meta-analysis has demonstrated that nesiritide may worsen renal function (RF) in patients with acute decompensated heart failure (ADHF). A nesiritide utilization evaluation (NUE) performed in our institution reflected that belief of RF preservation was one reason leading to nesiritide overuse. This study examined the effect of nesiritide on RF in patients evaluated in this NUE. RESEARCH DESIGN AND METHODS: Nesiritide patient records from October 1, 2003 to March 31, 2004 were reviewed (n = 162). Pertinent demographics, laboratory and medication utilization data were obtained. MAIN OUTCOME MEASURES: Changes in creatinine clearance (CrCl) and percentage of patients demonstrating worsening RF (decrease in CrCl > or = 25%) during diuretics therapy before nesiritide initiation and during nesiritide initiation and during nesiritide therapy were compared using Wilcoxon Signed-Ranks test and Chi-Square respectively. RESULTS: Overall, the addition of nesiritide to IV furosemide did not clinically change RF as compared to the use of IV furosemide alone (% change in CrCl: 0 vs. -2.13 mL/min, p = 0.023), although the difference was statistically significant. When categorized into different RF, there is no different in CrCl changes between the nesiritide group and the IV furosemide alone group. There is also no difference in terms of number of patients experiencing > or = 25% reduction in CrCl overall. Changes in RF were also not correlated to dose or duration of therapy. CONCLUSION: Nesiritide did not demonstrate significant impact in RF. Larger studies that examine RF in a more systematic and controlled manner and relate changes in RF to clinical outcomes are necessary to further elucidate the risk versus benefit profile of nesiritide. PMID- 16307708 TI - LDL-C/HDL-C ratio in subjects with cardiovascular disease and a low HDL-C: results of the RADAR (Rosuvastatin and Atorvastatin in different Dosages And Reverse cholesterol transport) study. AB - BACKGROUND: The ratio of low-density lipoprotein cholesterol and high-density lipoprotein cholesterol (LDL-C/HDL-C) is a reliable predictor of cardiovascular risk. Low HDL-C levels in patients with coronary artery disease are associated with a high risk for cardiovascular events. OBJECTIVES: This study compared the effects of rosuvastatin and atorvastatin on the LDL-C/HDL-C. METHODS: Patients aged 40-80 years with established cardiovascular disease and HDL-C < 1.0 mmol/L (< 40 mg/dL) entered as a 6-week dietary run-in period, before randomisation to open-label treatment with rosuvastatin 10 mg (n = 230) or atorvastatin 20 mg (n = 231) for 6 weeks. Doses were increased after 6 weeks to rosuvastatin 20 mg or atorvastatin 40 mg, and after 12 weeks to rosuvastatin 40 mg or atorvastatin 80 mg. Serum lipid parameters were measured at baseline and 6, 12 and 18 weeks. RESULTS: After 6 weeks of treatment, mean percentage change from baseline in LDL C/HDL-C ratio was -47.0% in the rosuvastatin group and -41.9% in the atorvastatin group (p < 0.05 for between-group comparison). After 12 and 18 weeks of treatment, change from baseline was -53.0% and -57.3%, respectively, for rosucastatin, compared with -47.9% and -49.6%, respectively, for atorvastatin (p < 0.01 and p < 0.001, respectively, for between-group comparison). Rosuvastatin also reduced LDL-C, total cholesterol/HDL-C significantly more than atorvastatin at all three time points, and significantly improved total cholesterol/HDL-C and apolipoprotein B/A-I ratios. CONCLUSIONS: Rosuvastatin 10, 20 and 40 mg is significantly more effective than atorvastatin 20, 40 and 80 mg, respectively, in improving the LDL-C/HDL-C ratio in patients with cardiovascular disease and low HDL-C. Further studies are required to clarify the benefits of rosuvastatin for reduction of cardiovascular risk. PMID- 16307709 TI - Analytic review of bimatoprost, latanoprost and travoprost in primary open angle glaucoma. AB - OBJECTIVE: The objective of this review was to evaluate different measures of efficacy of the intraocular pressure (IOP) lowering lipid class agents bimatoprost, latanoprost and travoprost in the treatment of primary open angle glaucoma. Study arms of timolol in trials including the above mentioned lipid class drugs were also included. METHODS: MEDLINE and EMBASE were searched for randomized clinical trials including one or more of the lipid class drugs bimatoprost, latanoprost and travoprost. The study results were pooled, and the simple, weighted IOP-lowering efficacy was compared among the lipid class drugs and timolol, where data were available. Efficacy parameters were reviewed, including mean reduction of IOP and percentage of patients achieving different levels of IOP. RESULTS: 161 articles were identified of which 42 were included in the analysis. A total of 9295 patients participated in the included trials. Based on all studies, timolol on average had a weighted mean IOP reduction of 22.2%, while latanoprost, travoprost and bimatoprost had a weighted mean IOP reduction of 26.7%, 28.7% and 30.3%, respectively. Analysis of target achievement to various IOP levels shows that bimatoprost seems more efficacious than latanoprost. The direct comparisons (head-to-head studies) also show that bimatoprost is the most efficacious treatment, however it is not conclusive whether latanoprost or travoprost is better in reducing IOP. CONCLUSIONS: This review shows that bimatoprost seems to be the most efficacious treatment in lowering IOP. Head-to-head studies confirm this. PMID- 16307710 TI - The effects of orlistat in patients with diabetes: improvement in glycaemic control and weight loss. AB - OBJECTIVE: In addition to direct weight reduction, there may be other benefits of obesity treatment including improved insulin sensitivity. The purpose of this study was to characterise concomitant diabetes drug use and the related costs in patients with diabetes treated with orlistat (Xenical) in the first 6 months of treatment. METHODS: One hundred overweight patients with diabetes and a body mass index (BMI) > or = 28 kg/m2 were enrolled in a structured UK hospital-based weight management clinic and treated with orlistat plus behavioural interventions. Among other measures, weight, glucose control (HbA1c) and drug treatment were recorded. Subjects were followed-up for a maximum of 24 months at intervals of 1-3 months, with a maximum treatment period of 24 months. RESULTS: The majority of subjects (91%) had type 2 diabetes. They had a mean age of 55 years and 55% were women. For patients followed up at 6 months, their mean BMI at baseline was 39.5 kg/m2 with a mean HbA1c of 7.6%. The mean weight loss at 6 months was 7.1 kg (p < 0.001). Despite a significant average absolute HbA1c reduction of 0.62% (p < 0.001), the most notable gains were made by those with the highest baseline HbA1c values (a mean relative reduction of 20% for those above the 75th percentile). There were 50 patients treated with insulin at baseline and 47 at 6 months. Of those treated with insulin, the mean dose was 130 units at baseline and 90 units at 6 months (p < 0.001). Twenty patients (44.4%) initially treated with oral hypoglycaemic agents alone reduced their dose after 6 months (not significant). Despite marked improvement in insulin sensitivity (baseline mean, 1.24 units/kg; 6 month mean, 0.90 units/kg [p < 0.001]) there was no correlation with BMI change. The average cost of diabetes treatment at baseline was pound 1.16 per day and pound 0.83 at 6 months (p < 0.001). Age was the only independent predictor for insulin dose reduction. CONCLUSIONS: Orlistat appears to reduce the need for concomitant diabetes medication irrespective of weight loss, a reduction that is likely to represent a large cost offset for orlistat treatment. PMID- 16307711 TI - The impact of secondary tasks on multi-tasking in a virtual environment. AB - One experiment is described that examined the possible involvement of working memory in the Virtual Errands Test (McGeorge et al. (2001). Using virtual environments in the assessment of executive dysfunction. Presence, 10, 375-383), which requires participants to complete errands within a virtual environment, presented on a computer screen. Time was limited, therefore participants had to swap between tasks (multi-task) efficiently to complete the errands. Forty-two undergraduates participated, all attempting the test twice. On one of these occasions they were asked to perform a concurrent task throughout (order of single and dual-task conditions was counterbalanced). The type of secondary task was manipulated between groups. Twenty-one participants were asked to randomly generate months of the year aloud in the dual-task condition, while another 21 were asked to suppress articulation by repeating the word "December". An overall dual-task effect on the Virtual Errands Test was observed, although this was qualified by an interaction with the order of single and dual-task conditions. Analysis of the secondary task data showed a drop in performance (relative to baseline) under dual-task conditions, and that drop was greater for the random generation group than the articulatory suppression group. These data are interpreted as suggesting that the central executive and phonological loop components of working memory are implicated in this test of multi-tasking. PMID- 16307712 TI - The interaction between response effects during the acquisition of response priming. AB - In three experiments we investigated the role of associative learning in the acquisition of response priming by effect stimuli, by examining their interaction during response-effect learning. Having replicated the ability of visual effect stimuli to prime their associated responses, we paired a response with a compound consisting of visual and auditory effects before assessing the ability of the auditory effect stimulus to prime the response. This priming was reduced if the visual stimulus had been pre-trained as an effect of the response. By contrast, priming by the visual effect stimulus was potentiated when the auditory effect had been pre-trained. We interpret these interactions in terms of contemporary associative learning theory derived from studies of conditioning. PMID- 16307713 TI - Monte Carlo simulation of colloidal membrane filtration: principal issues for modeling. AB - The principal issues involved in developing a Monte Carlo simulation model of colloidal membrane filtration are investigated in this study. An important object for modeling is the physical dynamics responsible for causing particle deposition and accumulation when encountering an open system with continuous outflow. A periodic boundary condition offers a solution to the problem by recirculating continuous flow back through the system. Scaling to full physical dimensions will allow for release of the model from flawed assumptions such as constant cake layer volume fraction and thickness throughout the system. Furthermore, rigorous modeling on a precise scale extends the model to account for random particle collisions with acute accuracy. A major finding of this study proves that forces within the colloidal filtration system are summed and transferred cumulatively through the inter-particle interactions. The force summation and transfer phenomenon only realizes its true value when the model is scaled to full dimensions. The overall strategy for model development, therefore, entails three stages: first, rigorous modeling on a microscopic scale; next, comprehensive inclusion of relevant physical dynamics; and finally, scaling to full physical dimensions. PMID- 16307714 TI - An assay for the determination of biologically active bone morphogenetic proteins using cells transfected with an inhibitor of differentiation promoter-luciferase construct. AB - Bone morphogenetic proteins (BMPs) control cell fate by regulating gene expression, especially inhibitor of differentiation (Id) genes. This property has been exploited to create a highly sensitive assay for quantification of active BMP. Embryonic mouse cells (C3H10T1/2) were stably transfected with an expression construct (BRE-Luc) containing a BMP-responsive element fused to the firefly luciferase reporter gene. BRE results from a multimerization of distinct sequences elements from a mouse Id1 promoter [15]. The addition of BMP-2 (0.5 100ng/ml) to the transfectants resulted in a dose-dependent increase in luciferase activity in the cell lysates. This new assay was 100-fold more sensitive than the classical alkaline phosphatase (ALP) activity assay (0.5-1 vs. 50-100ng/ml, respectively) as well as much more rapid (24h vs. 3-6 days, respectively, of BMP treatment). This new assay is specific to BMPs (BMP-2, BMP 4, and BMP7) as evidenced by its relative insensitivity to TGFbeta1, bFGF, and VEGF. Because of its BMP specificity, this rapid, sensitive, nonradioactive, and easily performed assay could be used in monitoring the biological activity of BMP and, eventually, as a cell-based screening assay to identify and evaluate molecules that modulate BMP signaling in cells. PMID- 16307715 TI - Preparation of caged compounds using an antibody against the photocleavable protecting group. PMID- 16307716 TI - Profiling of structurally labile oxylipins in plants by in situ derivatization with pentafluorobenzyl hydroxylamine. AB - A GC-MS-based method for the simultaneous quantification of common oxylipins along with labile and highly reactive compounds based on in situ derivatization with pentafluorobenzyl hydroxylamine to the corresponding O-2,3,4,5,6 pentafluorobenzyl oximes (PFB oximes) is presented. The approach covers oxo derivatives such as jasmonic acid (JA), 12-oxophytodienoic acid (OPDA), certain phytoprostanes, unsaturated oxo-acids, oxo-hydroxy acids, and aldehyde fragments from the polar head of fatty acids. In the positive electron impact-MS mode, the PFB oximes display characteristic fragment ions that greatly facilitate the identification of oxylipins in complex matrices. In addition, the fluorinated derivatives allow a highly selective and low-background analysis by negative chemical ionization. Besides showing the general value of the method for the identification of a broad range of oxylipins (18 examples), we also demonstrate sensitivity, linearity, and reproducibility for the quantification of JA, OPDA, 11-oxo-9-undecenoic acid, and 13-oxo-9,11-tridecadienoic acid. The efficiency of the method is demonstrated by differential profiling of these four oxylipins in lima bean leaves after mechanical wounding and feeding by the herbivore Spodoptera littoralis. Caterpillar feeding induced several oxylipins, whereas after wounding only the level of JA increased. The rapid in situ derivatization prevents the isomerization of cis-JA to trans-JA. The resting level of JA in lima beans showed an isomer ratio of 80:20 for trans/cis-JA. After wounding, de novo synthesis of JA alters the ratio to 20:80 in favor of the cis isomer. PMID- 16307717 TI - Comparative modeling and analysis of microfluidic and conventional DNA microarrays. AB - A theoretical analysis was developed to predict molecular hybridization rates for microarrays where samples flow through microfluidic channels and for conventional microarrays where samples remain stationary during hybridization. The theory was validated by using a multiplexed microfluidic microarray where eight samples were hybridized simultaneously against eight probes using 60-mer DNA strands. Mass transfer coefficients ranged over three orders of magnitude where either kinetic reaction rates or molecular diffusion rates controlled overall hybridization rates. Probes were printed using microfluidic channels and also conventional spotting techniques. Consistent with the theoretical model, the microfluidic microarray demonstrated the ability to print DNA probes in less than 1 min and to detect 10-pM target concentrations with hybridization times in less than 5 min. PMID- 16307718 TI - Protein/RNA coextraction and small two-dimensional polyacrylamide gel electrophoresis for proteomic/gene expression analysis of renal cancer biopsies. AB - A small amount of bioptic tissue ( approximately 5-10mg of fresh tissue) usually does not contain enough material to extract protein and RNA separately, to obtain preparative two-dimensional polyacrylamide gel electrophoresis (2-DE), and to identify a large number of separated proteins by MS. We tested a method, on small renal cancer specimens, for the coextraction of protein and RNA coupled with 2-DE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) or quadrupole time-of-flight (Q-TOF) analysis. We coextracted 0.28+/-0.05mg of proteins and 2.5+/-0.33microg of RNA for each 10mg of renal carcinoma tissue. Small and large 2-DE gels were compared: they showed a similar number of spots, and it was possible to match each other; using small format gels, one-fifth of the protein amount was required to identify, by Q-TOF analysis, the same number of proteins identifiable in large-format gel using MALDI-TOF analysis. Quality of RNA coextracted with the proteins was tested by real-time PCR on a set of housekeeping genes. They were quantified with high amplification efficiency and specificity. In conclusion, using 5 to 10mg of fresh tissue, it was possible to perform comprehensive parallel proteomic and genomic analysis by high-resolution, small-format 2-DE gels, allowing approximately 300 proteins identification and 1000 genes expression analysis. PMID- 16307719 TI - Binding efficiency of elongation factor Tu to tRNAs charged with nonnatural fluorescent amino acids. PMID- 16307720 TI - Quantification of the concentration and 13C tracer enrichment of long-chain fatty acyl-coenzyme A in muscle by liquid chromatography/mass spectrometry. AB - Recent diabetes and obesity research has been focused on the role of intracellular lipids in insulin resistance. Fatty acyl-coenzyme A (CoA) esters play a central role in the trafficking of intracellular lipids, but there has not previously been a method with which to quantify their kinetics using tracer methodology. We have therefore developed a high-performance liquid chromatography (HPLC)-mass spectrometry method to simultaneously measure the (13)C stable isotopic enrichment of palmitoyl-acyl-CoA ester and the concentrations of five individual long-chain fatty acyl-CoA esters extracted from muscle tissue samples. The long-chain fatty acyl-CoA can be effectively extracted from frozen muscle tissue samples and baseline separated by a reverse-phase HPLC with the presence of a volatile reagent-triethylamine. Negative ion electrospray mass spectrometry with selected ion monitoring was used to analyze the fatty acyl-CoAs to achieve reliable quantification of their concentrations and (13)C isotopic enrichment. Applying this protocol to rabbit muscle samples demonstrates that it is a sensitive, accurate, and precise method for the quantification of long-chain fatty acyl-CoA concentrations and enrichment. PMID- 16307721 TI - A nonterminal method for frequent collection of mouse circulating proteins by peritoneal lavage. PMID- 16307722 TI - Monitoring farnesol-induced toxicity in tobacco BY-2 cells with a fluorescent analog. AB - In a previous study (A. Hemmerlin, T.J. Bach, Plant Physiol. 123 (2000) 1257 1268), we have demonstrated that above a critical concentration, treatment with all-trans-farnesol induces cell-death in Nicotiana tabacum L. cv Bright Yellow-2 (TBY-2) cells. Now we used a fluorescent analog of farnesol (Fol(FLUO)), in which an isoprene unit is replaced by the fluorochrome 7-nitrobenz-2-oxa-1,3-diazol-4 yl, to visualize how cell integrity is affected. Fol(FLUO) exhibited the same toxicity as the natural compound and was shown to be readily taken up by TBY-2 cells, followed by integration into subcellular membrane structures. Although the plasma membrane seemed not to be labeled, Fol(FLUO) was associated with the tonoplast, endoplasmic reticulum, and Golgi apparatus or lipid bodies. Longer exposure times and increased Fol(FLUO) accumulation triggered the formation and proliferation of new membrane structures of as yet unknown function. Finally, at even higher and clearly cytotoxic concentrations of the analog, the cell contents became clearly disorganized, with cell swelling and ultimately plasmolysis. PMID- 16307723 TI - Effect of iron on bovine enamel and on the composition of the dental biofilm formed "in situ". AB - OBJECTIVES: This study investigated in situ the effect of iron (Fe) on the reduction of demineralization of bovine enamel, as well as on the composition of dental biofilm. DESIGN AND METHODS: Twelve volunteers were included in this blind crossover study, which was conducted in two stages of 14 days each. For each stage, the volunteers received palatal appliances containing four blocks of bovine enamel (4 mm x4 mm x 2.5 mm). Six volunteers dripped a solution of 15 mmol L(-1) ferrous sulphate onto the fragments and the remaining six dripped deionized water (eight times per day). After five minutes, a fresh 20% (w/v) sucrose solution was dripped onto all enamel blocks. During the experimental period the volunteers brushed their teeth with non-fluoridated dentifrice. After each stage, the percentage of surface microhardness change (%SMHC) and area of mineral loss (DeltaZ) were determined on enamel and the dental biofilm formed on the blocks was collected and analysed for F, P, Ca, Fe and alkali-soluble carbohydrates. The concentrations of F, Ca and Fe in enamel were also analysed after acid biopsies. RESULTS: There was a statistically significant increase in the P and Fe concentrations in the biofilms treated with ferrous sulphate (p<0.05), which was not observed for F, Ca and alkali-soluble carbohydrates. The group treated with ferrous sulphate had significantly lower %SMHC and DeltaZ when compared to control (p<0.05). CONCLUSIONS: These results showed that ferrous sulphate reduced the demineralization of enamel blocks and altered the ionic composition of the dental biofilm formed in situ. PMID- 16307724 TI - Functionalized carbon nanotubes as emerging nanovectors for the delivery of therapeutics. AB - Functionalized carbon nanotubes (f-CNT) are emerging as a new family of nanovectors for the delivery of different types of therapeutic molecules. The application of CNT in the field of carrier-mediated delivery has become possible after the recent discovery of their capacity to penetrate into the cells. CNT can be loaded with active molecules by forming stable covalent bonds or supramolecular assemblies based on noncovalent interactions. Once the cargos are carried into various cells, tissues and organs they are able to express their biological function. In this review, we will describe the potential of f-CNT to deliver different types of therapeutic molecules. PMID- 16307725 TI - Stimulation of catecholamine synthesis through unique estrogen receptors in the bovine adrenomedullary plasma membrane by 17beta-estradiol. AB - Incubation of cultured bovine adrenal medullary cells with 17beta-estradiol (E(2)) (0.3-100nM) or membrane-impermeable E(2)-bovine serum albumin (100nM) acutely increased (14)C-catecholamine synthesis from [(14)C]tyrosine. The stimulatory effect of E(2) was not inhibited by ICI182,780, a nuclear estrogen receptor inhibitor. E(2) also increased tyrosine hydroxylase activity and p44/42MAPK phosphorylation, the former of which was attenuated by U0126, an inhibitor of p44/42MAPK kinase. The plasma membrane isolated from the gland showed two classes of specific binding sites of [(3)H]E(2) with apparent K(d)s of 3.2 and 106nM, and B(max)s of 0.44 and 8.5pmol/mg protein, respectively. The high affinity binding of [(3)H]E(2) was most strongly inhibited by E(2) and phytoestrogens, and to lesser extents by other steroid hormones, while it was enhanced by ICI182,780 and environmental estrogenic pollutants. These findings suggest that E(2) acutely stimulates catecholamine synthesis via activation of p44/42MAPK through unique estrogen receptors in the plasma membrane of bovine adrenal medulla. PMID- 16307726 TI - Splice variant-specific silencing of angiotensin II type 1a receptor messenger RNA by RNA interference in vascular smooth muscle cells. AB - In the rat, two distinct angiotensin II type 1a (rAT(1a)) receptor mRNAs are synthesized from a single rAT(1a) receptor gene by alternative splicing. These two transcripts are comprised of exons 1, 2, and 3 (E1,2,3) or exons 1 and 3 (E1,3). Since exon 3 contains the entire coding region, both transcripts encode identical rAT(1a) receptors. Real-time PCR revealed that in rat aortic smooth muscle cells (RASMC), E1,2,3 mRNA accounted for 69.5+/-0.9% of total rAT(1a) receptor mRNA. The aim of this study was to use RNA interference (RNAi) to selectively silence the rAT(1a) receptor splice variants. Forty-eight hour treatment of RASMC with E1,3-targeting siRNA (10nM; S1(E1,3)) resulted in a 91.2+/-0.5% (n=3, P<0.001) reduction in E1,3 mRNA and a 19.0+/-3.0% (n=4, P<0.05) reduction in AT(1) receptor specific binding compared with cells treated with a non-silencing control siRNA; under these conditions, no effect was observed on levels of E1,2,3 mRNA. Conversely, treatment with E1,2,3-targeting siRNA (S2(E2)) had no effect on E1,3 mRNA while reducing E1,2,3 mRNA by 73.9+/-4.2% (n=3, P<0.001), and AT(1) receptor binding by 39.4+/-5.4% (n=4, P<0.001) compared with control. These data show that the majority of functional AT(1) receptor expression in RASMC derives from the E1,2,3 splice variant. These data also demonstrate that rAT(1a) receptor mRNA can be silenced in a splice-variant specific manner using siRNA in RASMC, thus providing an excellent model system for investigating the role of alternative splicing in the regulation of rAT(1a) receptor expression. PMID- 16307727 TI - tRNA-isoleucine-tryptophan composite gene. AB - Transfer-RNA genes in archaea often have introns intervening between exon sequences. The structural motif at the boundary between exon and intron is the bulge-helix-bulge. Computational investigations of these boundary structures in Haloarcula marismortui lead us to propose that tRNA-isoleucine and tRNA tryptophan genes are co-located. Precise in silico identification of the splice sites on the bulges at the exon-intron boundaries lead us to infer that a single intron-containing composite tRNA-gene can give rise to more than one gene product. PMID- 16307728 TI - BARHL1 homeogene, the human ortholog of the mouse Barhl1 involved in cerebellum development, shows regional and cellular specificities in restricted domains of developing human central nervous system. AB - The mouse homeobox gene Barhl1 plays a central role in cerebellum development and its expression is activated by the transcription factor Math1 which is involved in bone morphogenetic protein response pathways. We studied the human ortholog BARHL1 and we found that human, mouse, monkey, rat, and zebrafish orthologs were highly conserved and are members of the BarH homeogene family, containing Drosophila BarH1 and BarH2. The N-terminus of BARHL1 protein presents two FIL domains and an acidic domain rich in serine/threonine and proline, while the C terminus contains a canonical proline-rich domain. Secondary structure analysis showed that outside the three helixes of the homeodomain, BARHL1 protein has essentially random coil structure. We isolated BARHL1 and defined its expression pattern in human embryonic and fetal central nervous system (CNS) and compared it to the mouse Barhl1 transcription. BARHL1 mRNA was found exclusively in the CNS restricted to p1-p4 prosomeres of the diencephalon, to the dorsal cells of the mesencephalon, to the dorsal dl1 sensory neurons of the spinal cord, and to the rhombic lips yielding the cerebellar anlage. Detailed analysis of BARHL1 expression in fetal cerebellar cell layers using our new optic microscopy technology showed BARHL1 expression in external and internal granular cells and also in mouse adult granular cells, in agreement to Barhl1 null mouse phenotype affecting the differentiation and migration of granular cells. These findings indicate that the regional and cellular specificities of BARHL1 transcriptional control well correspond to the mouse Barhl1 transcription and suggest a potential role of this gene in the differentiation of BARHL1-expressing neuronal progenitors involved in the pattern formation of human cerebral and cerebellar structures. PMID- 16307729 TI - Crystal structure of the N-terminal SH3 domain of mouse betaPIX, p21-activated kinase-interacting exchange factor. AB - The mouse betaPIX-SH3 domain, residues 8-63 of P21-activated kinase interacting exchange factor, has been characterized by X-ray diffraction. Crystals belonging to space group P3(2)21 diffracted to 2.0 A and the structure was phased by the single-wavelength anomalous diffraction method. The domain is a compact beta barrel with an overall conformation similar to the general SH3 structure. The X ray structure shows mouse betaPIX-SH3 domain binding the way in which the betaPIX characteristic amino acids do so for an unconventional ligand binding surface. This arrangement provides a rationale for the unusual ligand recognition motif exhibited by mouse betaPIX-SH3 domain. Comparison with another SH3/peptide complex shows that the recognition mode of the mouse betaPIX-SH3 domain should be very similar to the RXXK ligand binding mode. The unique large and planar hydrophobic pocket may contribute to the promiscuity of betaPIX-SH3 domain resulting in its multiple biological functions. PMID- 16307730 TI - Nitric oxide scavenging by Mycobacterium leprae GlbO involves the formation of the ferric heme-bound peroxynitrite intermediate. AB - Ferrous oxygenated (Fe(II)O2) hemoglobins (Hb's) and myoglobins (Mb's) have been shown to react very rapidly with NO, yielding NO3(-) and the ferric heme-protein derivative (Fe(III)), by means of the ferric heme-bound peroxynitrite intermediate (Fe(III)OONO), according to the minimum reaction scheme: Fe(II)O2 + NO (k(on))--> Fe(III)OONO (h)--> Fe(III) + NO3(-). For most Hb's and Mb's, the first step (indicated by k(on)) is rate limiting, the overall reaction following a bimolecular behavior. By contrast, the rate of isomerization and dissociation of Fe(III)OONO (indicated by h) is rate limiting in NO scavenging by Fe(II)O2 murine neuroglobin, thus the overall reaction follows a monomolecular behavior. Here, we report the characterization of the NO scavenging reaction by Fe(II)O2 truncated Hb GlbO from Mycobacterium leprae. Values of k(on) (=2.1x10(6) M(-1) s( 1)) and h (=3.4 s(-1)) for NO scavenging by Fe(II)O2 M. leprae GlbO have been determined at pH 7.3 and 20.0 degrees C, the rate of Fe(III)OONO decay (h) is rate limiting. The Fe(III)OONO intermediate has been characterized by optical absorption spectroscopy in the Soret region. These results have been analyzed in parallel with those of monomeric and tetrameric globins as well as of flavoHb and discussed with regard to the three-dimensional structure of mycobacterial truncated Hbs and their proposed role in protection from nitrosative stress. PMID- 16307731 TI - Patterns of hippocampal cell loss based on subregional lesions of the hippocampus. AB - It is widely accepted that the hippocampus plays an essential role in memory. Furthermore, studies have suggested that subregions within the hippocampus contribute differentially to specific behavioral components of memory. These studies typically rely on lesions produced by localized injections of neurotoxins (e.g., ibotenic acid or colchicine) into targeted subregions of the hippocampus. In the present study, the specificity of ibotenic acid lesions into areas CA1 and CA3 and colchicine lesions into the dorsal dentate gyrus (DG) was tested. Specifically, the effects of lesions within the dorsal hippocampus, the ventral hippocampus, and areas outside the hippocampus (e.g., lateral septum and entorhinal cortex) were evaluated using Fluoro-Jade, a histofluorescent stain for degenerating neurons. The results show that cell loss is relatively uniform after ibotenic acid injections into areas CA1 and CA3 and variable after colchicine injections into DG. CA1 and CA3 lesions appeared mostly localized to those relative subregions, and DG lesions appeared highly localized to the DG. Using these lesion procedures, little cell loss was apparent in the ventral hippocampus, and no cell loss was apparent in the entorhinal cortex. It is suggested that the lesion procedures described in this study produce relatively selective lesions of neurons within specific subregions of the hippocampus and should be useful for studies examining possible differential contributions of hippocampal subregions to memory processes. PMID- 16307733 TI - Stereoselective single-step synthesis and X-ray crystallographic investigation of acetylated aryl 1,2-trans glycopyranosides and aryl 1,2-cis C2-hydroxy glycopyranosides. AB - Reported is an attractive and environmentally friendly method for the synthesis of the title compounds in moderate yield using inexpensive 1,2,3,4,6-penta-O acetyl-beta-D-gluco- and galactopyranoses as sugar donors, five different phenols as acceptors and H-beta zeolite as the catalyst. The yield (23-28%) of aryl 3,4,6 tri-O-acetyl-alpha-D-glycopyranosides obtained in this single-step procedure is considerably higher than that obtained using previously reported methods. Treatment of an orthoacetate, 3,4,6-tri-O-acetyl-[1,2-O-(1-p fluorophenoxyethylidene)]-alpha-D-glucopyranose, with p-fluorophenol under the same solvent-free reaction conditions also led to the formation of the title compounds in similar yield and composition. X-ray crystallographic analysis of phenyl 3,4,6-tri-O-acetyl-alpha-D-glucopyranoside and p-fluorophenyl 3,4,6-tri-O acetyl-alpha-D-glucopyranoside showed that the molecular packing is stabilized by C-H...O, C-H...pi and C-H...F interactions, in addition to regular hydrogen bonding patterns. PMID- 16307734 TI - Cardiac lipoprotein lipase: metabolic basis for diabetic heart disease. AB - The heart has a limited potential to synthesize fatty acid (FA), and, therefore, FA is supplied from several sources: lipolysis of endogenous cardiac triglyceride (TG) stores or from exogenous sources in the blood. Lipoprotein lipase (LPL), synthesized in cardiomyocytes, catalyzes the breakdown of the TG component of lipoproteins to provide FA to the heart. It is the vascular endothelial-bound LPL that determines the rate of plasma TG clearance, and, hence, it is also called heparin-releasable (HR) "functional" LPL. Functional LPL is regulated by numerous dietary and hormonal factors and is sensitive to pathophysiological alterations like those observed during diabetes. In this condition, absolute or relative lack of insulin impairs cardiac glucose transport and oxidation, resulting in FA becoming the preferred means of energy supply. To make available this increased requirement of the heart for FA, the diabetic heart upregulates its luminal LPL activity by posttranslational mechanisms. Chronically elevated cardiac LPL can result in abnormal FA supply and utilization by the heart tissue that could potentially initiate and sustain cardiac dysfunction during diabetes. As effective blood glucose control is difficult during diabetes, it is conceivable that a parallel increase in functional cardiac LPL activity may predispose people with diabetes to premature death from cardiac disease. By gaining more insight into the initial metabolic processes in the diabetic heart, we can attempt to piece together a part of the cascade of events leading to diabetic heart disease. PMID- 16307735 TI - Regulation of expression of Vg and establishment of the dorsoventral compartment boundary in the wing imaginal disc by Suppressor of Hairless. AB - The transcription factor Suppressor of Hairless (Su(H)) belongs to the CSL transcription factor family, which are the main transcriptional effectors of the Notch-signaling pathway. Su(H) is the only family member in the Drosophila genome and should therefore be the main transcriptional effector of the Notch pathway in this species. Despite this fact, in many developmental situations, the phenotype caused by loss of function of Su(H) is too weak for a factor that is supposed to mediate most or all aspects of Notch signaling. One example is the Su(H) mutant phenotype during the development of the wing, which is weaker in comparison to other genes required for Notch signaling. Another example is the complete absence of a phenotype upon loss of Su(H) function during the formation of the dorsoventral (D/V) compartment boundary, although the Notch pathway is required for this process. Recent work has shown that Su(H)/CBF1 has a second function as a transcriptional repressor, in the absence of the activity of the Notch pathway. As a repressor, Su(H) acts in a complex together with Hairless (H), which acts as a bridge to recruit the co-repressors Groucho and CtBP, and acts in a Notch independent manner to prevent the transcription of target genes. This raises the possibility that a de-repression of target genes can occur in the case of loss if function of Su(H). Here, we show that the weak phenotype of Su(H) mutants during wing development and the absence of a phenotype during formation of the D/V compartment boundary are caused by the concomitant loss of the Notch-independent repressor function. This loss of the repressor function of Su(H) results in a de repression of expression of target genes to a different degree in each process. Loss of Su(H) function during wing development results in a transient de repression of expression of the selector gene vestigial (vg). We show that this residual expression of vg is responsible for the weaker mutant phenotype of Su(H) in the wing. During the formation of the D/V compartment boundary, de-repression of target genes seems to be sufficiently strong, to compensate the loss of Su(H) activity. Thus, de-repression of its target genes obscures the involvement of Su(H) in this process. Furthermore, we provide evidence that Dx does not signal in a Su(H)-independent manner as has been suggested previously. PMID- 16307736 TI - Identification of a BMP inhibitor-responsive promoter module required for expression of the early neural gene zic1. AB - Expression of the transcription factor zic1 at the onset of gastrulation is one of the earliest molecular indicators of neural fate determination in Xenopus. Inhibition of bone morphogenetic protein (BMP) signaling is critical for activation of zic1 expression and fundamental for establishing neural identity in both vertebrates and invertebrates. The mechanism by which interruption of BMP signaling activates neural-specific gene expression is not understood. Here, we report identification of a 215 bp genomic module that is both necessary and sufficient to activate Xenopus zic1 transcription upon interruption of BMP signaling. Transgenic analyses demonstrate that this BMP inhibitory response module (BIRM) is required for expression in the whole embryo. Multiple consensus binding sites for specific transcription factor families within the BIRM are required for its activity and some of these regions are phylogenetically conserved between orthologous vertebrate zic1 genes. These data suggest that interruption of BMP signaling facilitates neural determination via a complex mechanism, involving multiple regulatory factors that cooperate to control zic1 expression. PMID- 16307737 TI - Training does not affect the alteration in pulmonary artery vasoreactivity in pulmonary hypertensive rats. AB - This study examined the effects of training on intrinsic vasorelaxation and vasoconstriction properties of pulmonary hypertensive rat arteries. Fifty seven male Wistar rats were randomly assigned to 4 groups: normotensive sedentary (n = 14), normotensive trained (n = 15), pulmonary hypertensive sedentary (n = 15) and pulmonary hypertensive trained (n = 13). Pulmonary hypertension was obtained using a chronic hypoxia exposure model. Endothelium-dependent vasorelaxation to acetylcholine (10(-8)-10(-4) M), endothelium-independent vasorelaxation to sodium nitro-prusside (10(-8)-10(-4) M), and vasoconstriction to epinephrine (10(-9)-10( 4) M) and endothelin-1 (10(-12)-10(-7) M) were assessed on isolated rings of large pulmonary arteries. Alterations in endothelium-dependent and -independent vasorelaxation properties as well as enhanced vasoconstrictor responses were obtained in pulmonary hypertensive rats. Chronic exercise did not affect those pulmonary vasoreactivity alterations. A predominant effect of chronic hypoxia over training seems to be partially responsible for this phenomenon, probably through impairment in nitric oxide bioavailability and vascular smooth muscle sensitivity. PMID- 16307738 TI - Effects of fasudil, a Rho-kinase inhibitor, on myocardial preconditioning in anesthetized rats. AB - The aim of this study was to examine the effects of fasudil, a Rho-kinase inhibitor, on ischemic preconditioning and carbachol preconditioning in anesthetized rats. The total number of ventricular ectopic beats was markedly augmented with fasudil at 0.3 mg/kg and depressed with fasudil at 10 mg/kg. Fasudil at 10 mg/kg also markedly decreased the ventricular tachycardia incidence. Ischemic preconditioning, induced by 5 min coronary artery occlusion and 5 min reperfusion, decreased the incidence of ventricular tachycardia and abolished the occurrence of ventricular fibrillation. The incidences of ventricular tachycardia and ventricular fibrillation in the fasudil (10 mg/kg) + ischemic preconditioning group were found to be similar to the ischemic preconditioning group. However, low doses of fasudil (0.3 and 1 mg/kg) appeared to prevent the antiarrhythmic effects of ischemic preconditioning. Carbachol (4 microg/kg/min for 5 min) induced marked reductions in mean arterial blood pressure, heart rate and abolished ventricular tachycardia. Marked reductions in ventricular ectopic beats and ventricular tachycardia were noted in the fasudil (10 mg/kg) + carbachol preconditioning group. Lactate levels were markedly reduced in the ischemic preconditioning group and this reduction was prominently inhibited with fasudil at 1 mg/kg. Ischemic preconditioning caused a marked decrease in plasma malondialdehyde levels. Fasudil (10 mg/kg), ischemic preconditioning and carbachol preconditioning each generated marked reductions in ischemic myocardial malondialdehyde levels. Decreases in infarct size were observed with fasudil (10 mg/kg) treatment, ischemic preconditioning and carbachol preconditioning when compared to control. These results suggest that low doses of fasudil (0.3 and 1 mg/kg) appeared to prevents the effects of ischemic preconditioning and carbachol preconditioning, but a high dose of fasudil (10 mg/kg) was able to produce cardioprotective effects on myocardium against arrhythmias, infarct size or biochemical parameters and mimic the effects of ischemic preconditioning in anesthetized rats. PMID- 16307739 TI - Modulation of striatal dopamine release in vitro by agonists of the glycineB site of NMDA receptors; interaction with antipsychotics. AB - The N-methyl-D-aspartate (NMDA) glutamate receptor possesses an obligatory co agonist site for D-serine and glycine, named the glycineB site. Several clinical trials indicate that glycineB agonists can improve negative and cognitive symptoms of schizophrenia when co-administered with antipsychotics. In the present study we have investigated the effects of glycineB agonists on the endogenous release of dopamine from preparations of rat striatal tissue prisms in static conditions. The glycineB agonists glycine (1 mM) and D-serine (10 microM), but not D-cycloserine (10 microM), substantially increased the spontaneous release of dopamine, but significantly reduced the release of dopamine evoked by NMDA. The effect of glycine on spontaneous release was abolished by the non competitive NMDA antagonists 5R,10S-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d] cyclohepten-5,10-imine (MK-801, 10 microM) and ifenprodil (5 microM), but was only partially suppressed by the competitive antagonist 4-(3-phosphonopropyl) piperazine-2-carboxylic acid (CPP, 10 microM). The selective inhibitor of the glial glycine transporter GlyT1 N[3-(4'-fluorophenyl)-3-(4' phenylphenoxy)propyl]sarcosine (NFPS, 10 microM) significantly increased the release of dopamine in an MK-801-sensitive manner. Interestingly, haloperidol (1 microM), but not clozapine (10 microM), prevented the effects of glycine. This study shows that glycineB modulators can control dopamine release by interacting with a distinctive NMDA receptor subtype with which some typical antipsychotics can interfere. PMID- 16307740 TI - The selective tachykinin neurokinin 1 (NK1) receptor antagonist, GR 205,171, stereospecifically inhibits light-induced phase advances of hamster circadian activity rhythms. AB - Circadian rhythms in mammals are generated by master pacemaker cells located within the suprachiasmatic nucleus of the hypothalamus. In hamsters, the suprachiasmatic nucleus contains a small collection of cells immunoreactive for substance P, the endogenous ligand of tachykinin neurokinin 1 (NK1) receptors. In addition, two other nuclei which form part of the circadian system, the intergeniculate leaflet of the thalamus and the raphe nuclei, also contain fibers and/or cell bodies immunoreactive for substance P. In light of these observations, we evaluated the influence of the selective tachykinin NK1 receptor antagonist, GR 205,171, upon circadian activity rhythms in the hamster. Systemic injection of GR 205,171 dose-dependently (2.5-40.0 mg/kg, i.p.) inhibited light induced phase advances in hamster circadian wheel running activity rhythms by approximately 50%. In contrast, GR 226,206, the less active enantiomer of GR 205,171, failed to affect light-induced phase advances. In addition, we examined the potential ability of GR 205,171 to induce non-photic phase shifts in hamster wheel running rhythms when injected at mid-day to late night circadian times. However, GR 205,171 (40 mg/kg) did not elicit non-photic phase shifts at these times indicating that tachykinin NK1 receptor antagonists are only effective when a light stimulus is applied to the pacemaker. Although GR 205,171 may, in theory, activate several sites within the circadian system, we suggest that GR 205,171 acts in the raphe nuclei to increase inhibitory serotonergic input to pacemaker cells in the suprachiasmatic nuclei, thereby suppressing photic modulation of the pacemaker. These findings have important implications for the use of tachykinin NK1 receptor antagonists in the treatment of depression and other central nervous system disorders. PMID- 16307741 TI - Activation of c-Jun NH2-terminal kinase is required for gemcitabine's cytotoxic effect in human lung cancer H1299 cells. AB - Although gemcitabine is a potent therapeutic agent in the treatment of human non small cell lung cancer (NSCLC), resistance to gemcitabine is common. In this study, we investigated the molecular mechanisms involved in acquired gemcitabine resistance against NSCLC cells. Gemcitabine-resistant NSCLC H1299 cells (H1299/GR) were selected by long-term exposure of parental H1299 cells to gemcitabine. The median inhibitory concentrations of gemcitabine in H1299 and H1299/GR cells were 19.4 and 233.1 nM, respectively. Gemcitabine induced activation of c-Jun NH2-terminal kinase (JNK) in parental H1299 cells but not in H1299/GR cells after 48 h. Blocking JNK activation by pretreatment with SP600125, a specific JNK inhibitor, or by transfection with dominant-negative JNK vectors abrogated gemcitabine-induced apoptosis in parental H1299 cells as evidenced by interruption of caspase activation. Transient transfection with a JNKK2-JNK1 plasmid expressing constitutive JNK1 partially restored the effect of gemcitabine in H1299/GR cells. Our results indicate that gemcitabine-induced apoptosis in human NSCLC H1299 cells requires activation of the JNK signaling pathway. Attenuated JNK activation may contribute to development of acquired gemcitabine resistance in cancer cells. PMID- 16307742 TI - Identification of voltage-dependent Ca2+ channels in sea urchin sperm. AB - Functional evidence indicates that voltage-dependent Ca2+ (Cav) channels participate in sea urchin sperm motility and the acrosome reaction (AR), however, their molecular identity remains unknown. We have identified transcripts for two Ca2+ channel alpha1 subunits in sea urchin testis similar in sequence to Cav1.2 and Cav2.3. Antibodies against rat Cav1.2 and Cav2.3 channels differentially label proteins in the flagella and acrosome of mature sea urchin sperm. The Cav channel antagonists nifedipine and nimodipine, which inhibit the AR, diminish the intracellular Ca2+ elevation induced by a K+-induced depolarization in valinomycin-treated sperm. These findings reveal that Cav1.2 and Cav2.3 channels could participate in motility and/or the AR in sea urchin sperm. PMID- 16307743 TI - An all atom energy based computational protocol for predicting binding affinities of protein-ligand complexes. AB - We report here a computationally fast protocol for predicting binding affinities of non-metallo protein-ligand complexes. The protocol builds in an all atom energy based empirical scoring function comprising electrostatics, van der Waals, hydrophobicity and loss of conformational entropy of protein side chains upon ligand binding. The method is designed to ensure transferability across diverse systems and has been validated on a heterogenous dataset of 161 complexes consisting of 55 unique protein targets. The scoring function trained on a dataset of 61 complexes yielded a correlation of r=0.92 for the predicted binding free energies against the experimental binding affinities. Model validation and parameter analysis studies ensure the predictive ability of the scoring function. When tested on the remaining 100 protein-ligand complexes a correlation of r=0.92 was recovered. The high correlation obtained underscores the potential applicability of the methodology in drug design endeavors. The scoring function has been web enabled at as binding affinity prediction of protein-ligand (BAPPL) server. PMID- 16307744 TI - Cervical inflammatory cytokines and other markers in the cervical mucus of pregnant women with lower genital tract infection. AB - OBJECTIVE: To evaluate concentrations of interleukin (IL)-6, IL-8, lactoferrin (LF), and alpha defencine (alpha-DF) in the cervical mucus of pregnant women and analyze their relation to cervicitis and bacterial vaginosis (BV). METHODS: Cervical mucus samples were obtained from August 2003 through May 2004 from 157 women who were between the 6th and 36th week of an uncomplicated singleton pregnancy. All women were delivered at term, 69 without BV or cervicitis, 9 with BV, and 79 with cervicitis. RESULTS: Interleukin 8, LF, and mu-DF concentrations were higher in women with cervicitis (0.81 +/- 0.36 pg/mL, 14.8+/-12.3 microg/mL, and 0.60 +/- 0.49 microg/mL) than in women without BV or cervicitis (0.35 +/- 0.34 pg/mL, 8.0 +/- 11.0 microg/mL, and 0.15 +/- 0.12 microg/mL). Interleukin 6 concentration was higher in women with BV (0.26 +/- 0.32 pg/mL) than in women without BV or cervicitis (0.09 +/- 0.15 pg/mL) or in women with cervicitis (0.12 +/- 0.18 pg/mL). CONCLUSIONS: Higher levels of inflammatory cytokines in the cervical mucus of pregnant women may lead to early detection of lower genital tract infection. PMID- 16307745 TI - Microsatellite analysis reveals genetic structure of Leishmania tropica. AB - The current rapid spread of leishmaniases caused by Leishmania tropica and the complexity of its clinical spectrum call for this parasite's epidemiological and evolutionary investigation. Evaluation of its population structure by isoenzyme electrophoresis and previous molecular biological analysis has proved difficult. In this study, we used 21 microsatellite loci to type 117 strains from different African and Asian locations. Eighty-one different genotypes were found. A genetic bottleneck supported by a gradient in the number of alleles and consistent with the geographical structure of the Middle East suggests an African origin of this species. A Bayesian approach identified 10 genetic clusters that correlated predominantly with geographical origin. The strains in the 'Asia' cluster form a very heterogeneous sub-population, with a varied but inter-related genotype that is geographically very widely dispersed and consistent with anthroponotic transmission of the parasite. The other nine clusters were more homogenous. The propagation of L. tropica appears to be predominantly clonal. In Africa and the Middle East, anthroponotic and zoonotic systems of distribution may contribute to the development of overlapping, genetically distinct populations of L. tropica. PMID- 16307746 TI - ApoA-I/phosphatidylcholine discs remodels fast-migrating HDL into slow-migrating HDL as characterized by capillary isotachophoresis. AB - OBJECTIVE: Capillary isotachophoresis (cITP) is a technique for characterizing plasma lipoprotein subfractions according to their electrophoretic charges. We used this technique to examine the mechanism by which apoA-I/phosphatidylcholine (POPC) discs increase pre-beta HDL. METHODS AND RESULTS: The cITP analysis was performed using plasma prestained with a lipophilic dye on a Beckman P/ACE MDQ system. Plasma from a patient with lecithin:cholesterol acyltransferase (LCAT) deficiency who had increased apoE-containing HDL was used to characterize the charge distribution of apoA-I/POPC discs. cITP analysis of apoB- and E-depleted plasma of the patient in the presence of apoA-I/POPC discs indicated two major subfractions of apoA-I/POPC discs with mobilities of triglyceride-rich lipoproteins (fast and slow apoA-I). Incubation of whole plasma from a normolipidemic subject in the presence of apoA-I/POPC discs caused a reduction in cITP fast (f)- and intermediate (i)-migrating HDL, and fast and slow apoA-I, and an increase in slow (s)-migrating HDL. The changes in cITP lipoprotein subfractions were not affected by the inhibition of LCAT activity. ApoA-I/POPC discs increased the fractional esterification rate of cholesterol in apoB depleted plasma. CONCLUSION: ApoA-I/POPC discs remodeled cITP fHDL and iHDL to sHDL independent of LCAT activity. PMID- 16307747 TI - Relationship between serum adiponectin level and lipid composition in each lipoprotein fraction in adolescent children. AB - The association of lipid composition in each lipoprotein fraction with adiponectin level in relation to body fatness was investigated. The subjects were 283 children (144 boys and 139 girls) aged 11.6 years (S.D. 1.5). Cholesterol (C) and triglyceride (TG) levels in each lipoprotein fraction were measured by a combination of agarose gel electrophoresis and differential staining. Adiponectin level was not significantly different between girls and boys. In simple regression analyses, an association of adiponectin level with lipoprotein lipid profile was shown only in girls, and not in boys at all. In girls, very low density lipoprotein (VLDL)-C, low-density lipoprotein (LDL)-C, VLDL-TG and LDL-TG were negatively correlated with adiponectin level, and high-density lipoprotein (HDL)-C and HDL-C/TG ratio were positively correlated. Multiple regression analyses including adiponectin level and body mass index (BMI) as predictors for the lipoprotein lipid profile demonstrated that adiponectin level was a significant independent predictor of VLDL-TG, LDL-C, LDL-TG, HDL-C and HDL-C/TG ratio. In conclusion, adiponectin level correlates with lipoprotein lipid profile independent of body fatness in adolescent girls. PMID- 16307748 TI - Antibodies of IgM subclass to phosphorylcholine and oxidized LDL are protective factors for atherosclerosis in patients with hypertension. AB - OBJECTIVE: To determine the importance of antibodies against phosphorylcholine (PC) and oxidized low density lipoprotein (OxLDL) for development of atherosclerosis. METHODS AND RESULTS: Two hundred and twenty six individuals with established hypertension (diastolic pressure > 95mmHg) were from European Lacidipine Study on Atherosclerosis. Antibodies of IgG and IgM subclass were tested by ELISA against PC (aPC), cupper-oxidized (ox)- or malondialdehyde (MDA) modified LDL. High-sensitivity C-reactive protein was measured by nephelometry. As a surrogate measure of atherosclerosis, we used the mean of the maximum intima media thicknesses (IMT) in the far walls of common carotids and bifurcations was determined by ultrasonography at the time of enrolment, and 4 years following enrolment. aPC could be competed out by PC and OxLDL, while cardiolipin (CL) and beta2-glycoprotein I (beta2GPI) were less effective and phosphatidylserine (PS) not at all. Increases in IMT at follow-up were less common in subjects which at the time of enrolment had high IgM aPC (both 75th and 90th; odds ratios: 0.46; CI: 0.25-0.85; 0.36; CI: 0.15-0.87) and high IgM aOxLDL and aMDA-LDL (90th; odds ratios 0.27; p = 0.01; CI: 0.11-0.69 and 0.27; p = 0.01; CI: 0.11-0.69). CRP was unrelated to IMT-changes. The relationship between IgM aPC, aOxLDL and aMDA-LDL and changes in IMT was independent of age, treatment with atenolol or lacidipine, smoking and lipids. Women had higher levels of IgM antibodies tested (p < 0.05). CONCLUSIONS: High levels of IgM-antibodies against PC and OxLDL predict a favourable outcome in the development of carotid atherosclerosis in hypertensive subjects. Whether these antibodies could be used therapeutically deserves further study. PMID- 16307749 TI - Mathematical model that predicts lower leg motion in response to electrical stimulation. AB - Electrical stimulation of skeletal muscles of patients with upper motor neuron lesions can be used to restore functional movements such as standing or walking. Mathematical muscle models can assist in designing stimulation patterns that will enable patients to perform particular tasks more efficiently. In this study we extend our previous model to allow us to predict changes in knee joint angle in response to electrical stimulation of the human quadriceps femoris muscle. The model was tested both with and without inertial loads placed around the ankle joints of healthy subjects. Results showed that the model predicted the knee extensions with a RMS angle error that was generally 45 microm) fraction and their wash-off behavior was studied. The fine and coarse fraction showed different wash off behaviors. During the runoff the concentration of the fine fraction was less fluctuated compared to coarse. The fluctuation of the coarse fraction was more influenced by TSS concentration. The PAH content measurement in fine fraction showed less fluctuation compared to the coarse fraction. The PAH content in the coarse fraction was found decreasing with increasing the coarse fraction contribution to TSS. PMID- 16307773 TI - Feasibility of the silver-UV process for drinking water disinfection. AB - A synergistic effect between cationic silver and UV radiation (silver-UV disinfection) has been observed that can appreciably enhance inactivation of viruses. The purpose of this work was to assess the feasibility of this technique for drinking water disinfection and evaluate the effects of selected impurities, found in fresh water, and common parameters on inactivation of the coliphage MS-2 with the silver-UV process. Turbidity (kaolin), calcium hardness, carbonate alkalinity, and pH did not significantly degrade inactivation. Inactivation was reduced in the presence of chloride, at concentrations greater than 30 mg/L, and in water samples with UV-254 absorbance values greater than ca. 0.1 cm(-1). Inactivation of MS-2 with silver-UV disinfection was also reduced at high phosphate concentrations (above ca. 5 mM). Silver-UV inactivation of MS-2 increased with increases in temperature between 10 and 20 degrees C. Silver-UV inactivation of MS-2 was increased by greater than 1-log over UV alone, in two untreated fresh water sources, which indicates that silver-UV may be a viable treatment technology. An assessment of operation and management costs suggests that an increase in inactivation of MS-2 with silver-UV disinfection could be economically beneficial. PMID- 16307774 TI - Modeling sediment impact on the transport of fecal bacteria. AB - Sediment storage of fecal bacteria has been observed for a long time. However, limited modeling efforts have been conducted to dynamically and explicitly study the impact of sediment on the transport of fecal bacteria. In this paper, a modeling framework based on the Environmental Fluid Dynamics Code (EFDC) model has been proposed. Free and particle-associated fecal bacteria can be modeled explicitly using the developed model. A linear adsorption relationship is incorporated into the model to obtain a sediment concentration dependent attachment ratio. The deposition and resuspension fluxes of fecal bacteria across the sediment bed-water interface are calculated coupled with the sediment deposition and resuspension flux. The model was applied to hypothetical scenarios to evaluate the sediment impacts under ideal settling and resuspension conditions. A case study was provided to test the resuspension calculation of fecal bacteria under an artificial flooding condition. Using this model, the contributions of fecal bacteria from sediment bed and from the watershed can be modeled separately and explicitly. PMID- 16307775 TI - The fate of nitrogen in a moderately alkaline and calcareous soil amended with biosolids and urea. AB - The determination of nitrogen (N) based loading rates for land application of biosolids is challenging and site specific. Over loading may contribute to environmental, agricultural, or human health problems. The objective of this study was to monitor N mineralization and losses in a moderately alkaline and calcareous desert soil amended with either anaerobically digested (AN) or lime stabilized (LS) biosolids, and irrigated with and without urea enriched water. For Experiment 1, N inputs, leaching and residuals in soil were evaluated in an open soil column system. For Experiment 2, ammonia (NH(3)) emissions were evaluated in a closed soil column system. In Experiment 1, AN and LS biosolids increased soil ON (organic N) by three and two fold, respectively. Respective net N mineralization of ON from biosolids alone was 90% and 62% without urea, and 71% and 77%, respectively with added urea. Nitrogen leaching losses and residuals in amended soil did not account for all N inputs into the soil/biosolids system. In Experiment 2, NH(3) emissions were not significantly different among treated soils with or without added urea, except LS amended soil receiving urea. Ammonia losses did not account for unaccounted N in Experiment 1. We concluded that deep placement and rapid mineralization of AN biosolids promoted anaerobic soil conditions and denitrification, in addition to the high denitrification potential of desert soil. LS biosolids showed greater potential than AN biosolids for safe and beneficial land application to desert soils regardless of biosolids placement and the inclusion of N rich irrigation water. PMID- 16307776 TI - Adsorption of the quinolone antibiotic nalidixic acid onto anion-exchange and neutral polymers. AB - Pharmaceutical products are being found in surface and ground waters around the world. While the consequences to human health are unknown, it is suspected that these contaminants adversely alter aquatic ecosystems. This study presents adsorption results for one pharmaceutical product, nalidixic acid (NA), on neutral and anion-exchange polymers at three pH values. The adsorption of NA below and above its pKa of approximately 6 on polymers with different matrices, forms, and degrees of polarity were evaluated. By comparing the Freundlich constants KF and n, the results show that the neutral form of NA adsorbs to a greater extent on neutral polymers, and that the anionic form of NA adsorbs more to anion-exchange polymers. Also, results of the adsorption experiments suggest that aromatic ring interactions between NA and the surface of both neutral and anion-exchange polymers are important in the adsorption process. These results have important implications for the treatment of pharmaceutical-contaminated water, as many pharmaceutical contaminants are ionizable and have aromatic rings in their structure. This study illustrates the importance of pH and sorbate and sorbent structure in considering polymer sorption for treatment of pharmaceutical contaminated aqueous systems. PMID- 16307777 TI - Phytoextraction of metals from a multiply contaminated soil by Indian mustard. AB - The effects of nitrilotriacetate (NTA) and citric acid applications on metal extractability from a multiply metal-contaminated soil, as well as on their uptake and accumulation by Indian mustard (Brassica juncea) were investigated. Desorption of metals from the soil increased with chelate concentration, NTA being more effective than citric acid in solubilising the metals. Plants were grown in a sandy soil collected from a contaminated field site and polluted by Cd, Cr, Cu, Pb and Zn. After 43 days of plant growth, pots were amended with NTA or citric acid at 5 mmol kg-1 soil. Control pots were not treated with any chelate. Harvest of plants was performed 1 week after chelate addition. Soil water-, NH4NO3- and DTPA-extractable Cd, Cu, Pb and Zn fractions were enhanced only in the presence of NTA. In comparison to unamended plants, Indian mustard shoot dry weights suffered significant reductions following NTA application. NTA treatment increased shoot metal concentrations by a factor of 2-3, whereas citric acid did not induce any difference compared to the control. Chromium was detected in the above-ground tissues only after NTA amendment. Due to differences in dry matter yield, a significant enhancement of metal uptake was observed in NTA treated plants for Cu and Zn. PMID- 16307778 TI - Cyanide detoxification by recombinant bacterial rhodanese. AB - Cyanide is a major environmental pollutant of the chemical and metallurgical industries. Although extremely toxic, cyanide can enzymatically be converted to the less toxic thiocyanate by rhodaneses (thiosulfate:cyanide sulfurtransferases, EC 2.8.1.1). We engineered a genetic system to express high levels of recombinant Pseudomonas aeruginosa rhodanese (r-RhdA) in Escherichia coli, and used this organism to test the role of r-RhdA in cyanide detoxification. Inducible expression of the rhdA gene under the control of the hybrid T7-lacO promoter yielded active r-RhdA over a 4-h period, though r-RhdA-expressing E. coli showed decreased viability starting from 1 h post-induction. At this time, Western blot analysis and enzymatic assay showed r-RhdA partition between the cytoplasm (95%) and the periplasm (5%). The accessibility of thiosulfate to r-RhdA was a limiting step for the sulfur transfer reaction in the cellular system, but cyanide conversion to thiocyanate could be increased upon permeabilization of the bacterial membrane. Specific r-RhdA activity was higher in the whole-cell assay than in the in vitro assay with pure enzyme (2154 vs. 816 micromol min-1 mg-1 r RhdA, respectively), likely reflecting enzyme stability. The r-RhdA-dependent cyanide detoxification resulted in increased resistance of r-RhdA overexpressing E. coli to 5 mM cyanide. Bacterial survival was paralleled by release of thiocyanate into the medium. Our results indicate that cyanide detoxification by engineered E. coli cells is feasible under laboratory conditions, and suggest that microbial rhodaneses may contribute to cyanide transformation in natural environments. PMID- 16307779 TI - Effects of pH and electrolyte concentration on the binding between a humic acid and an oxazine dye. AB - The binding between an oxazine dye and a humic acid was studied in aqueous solutions in the pH range 4-10 and in the supporting electrolyte (KCl) range 0.001-0.1M. A rather simple spectrophotometric method was developed to construct binding isotherms under conditions were traditional centrifugation or filtration methods fail. The use of this method is possible because humic acid molecules have the ability of changing the spectrum of dye molecules, and this ability is used to quantify the isotherms. All binding isotherms have a Langmuirian shape. The amount of bound dye is strongly dependent on the ionic strength and less dependent on the pH of the solution. The binding is rather strong and mainly driven by non-electrostatic forces. Whereas the Langmuir binding constant is independent of the pH and electrolyte concentration, the number of assessable sites in humic acid for binding oxazine increases by increasing pH and decreasing electrolyte concentration. These results can be directly related to the flexibility of humic acid molecules, which can swell at high pH and low ionic strength, increasing consequently the availability of binding sites. The results also indicate that humic substances may strongly affect the mobility and fate of dyes and related pollutants in the environment. PMID- 16307780 TI - Eutrophication trends in forest soils in Galicia (NW Spain) caused by the atmospheric deposition of nitrogen compounds. AB - We calculated the sensitivity of Galician forest soils to eutrophication caused by atmospheric deposition of nitrogen compounds, using the Simple Mass Balance (SMB) method as described by [Posch, M., de Vries, W., Hettelingh, J.-P., 1995. Critical loads of sulphur and nitrogen. In: Posch, M., de Smet, P.A.M., Hettelingh, J.-P., Downing, R.J. Calculation and Mapping of Critical Thresholds in Europe. Status Report 1995, Coordination Center for Effects, National Institute for Public Health and the Environment, Bilthoven, The Netherlands, pp. 31-42]. Deposition values were used to calculate critical loads exceedance. Galician natural forest ecosystems can support nitrogen deposition loads of more than 10 kg Nha (-1) yr (-1). The lowest critical loads (approximately 10 kg Nha ( 1) yr (-1)) mainly occurred in forest stands in the interior zone, while highest critical load values (approximately 68 kg Nha (-1) yr (-1)) were observed in eucalyptus stands at low altitudes in the littoral area. Exceedances based on N deposition levels, calculated from data recorded in 2001, occurred in 40% of the forest soils, showing the need to control N emissions in these areas to prevent possible eutrophication of soils and waters. Analysis of rainfall bulk composition revealed that ammonium, probably derived from agricultural and cattle activities, was the main compound responsible for N deposition in Galicia. PMID- 16307781 TI - Effect of activated sludge properties and membrane operation conditions on fouling characteristics in membrane bioreactors. AB - Biofouling control is considered to be a major challenge in operating membrane bioreactors (MBRs) for the treatment of wastewater. This study examined the impact of biological, chemical, and physical properties of activated sludge on membrane filtration performance in laboratory-scale MBRs. Sludges with different microbial communities were produced using pseudo-continuous stirred-tank reactors and pseudo-plug flow reactors treating a synthetic paper mill wastewater. Various filtration resistances were used to investigate membrane fouling characteristics, and molecular biology tools targeting 16S ribosomal DNA gene sequences were used to identify predominant bacterial populations in the sludges or attached to the fouled membranes. Filtration experiments using axenic cultures of Escherichia coli, Acinetobacter calcoaceticus, and Gordonia amarae were also performed to better understand the initiation and development of biofouling. The results showed that the tendency of membranes to biofoul depended upon membrane operating conditions as well as the properties of the activated sludge in the MBR systems. Specific bacterial populations, which were not dominant in the activated sludges, were selectively accumulated on the membrane surface leading to the development of irreversible biofouling. PMID- 16307782 TI - Characterization of atmospheric particulate and metallic elements at Taichung Harbor near Taiwan Strait during 2004-2005. AB - Air aerosol samples for TSP (total suspended particulate), coarse particulate (particle matter with aerodynamical diameter 2.5-10 microm, PM(2.5-10)), fine particulate (particle matter with aerodynamical diameter <2.5 microm, PM(2.5)) and metallic elements were collected during March 2004 to January 2005 at TH (Taichung Harbor) in central Taiwan. The seasonal variation average concentration of TSP (total suspended particulate), coarse particulate (particle matter with aerodynamical diameter 2.5-10 microm, PM(2.5-10)) and fine particulate (particle matter with aerodynamical diameter <2.5 microm, PM(2.5)) were in the range 132 171.1 microg m(-3) and 43-49.5 microg m(-3), respectively. Seasonal variation of metallic elements Cu, Mn, Zn and Fe in the TSP (total suspended particulate) shows that higher concentration was observed during spring. Seasonal variation of metallic elements Pb, Cr and Mg in the TSP (total suspended particulate) shows that higher concentration was observed during winter. The average metallic element TSP (total suspended particulate) concentration order was Fe>Zn>Mg>Cu>Cr>Mn>Pb in spring. In addition, at the TH sampling site, the average concentration variation of TSP (total suspended particulate) displayed the following order: spring>winter>autumn>summer. However, the average concentration variation of coarse particulate (particle matter with aerodynamical diameter 2.5 10 microm, PM(2.5-10)) displayed the following order: spring>winter>summer>autumn. Finally, the average concentration variations of fine particulate (particle matter with aerodynamical diameter <2.5 microm, PM(2.5)) were in the following order: winter>spring>summer>autumn at the TH sample site. PMID- 16307783 TI - Evaluating spectroscopic and chromatographic techniques to resolve dissolved organic matter via end member mixing analysis. AB - Real-time or near real-time in-situ monitoring of dissolved organic matter (DOM) composition in natural waters and engineered treatment systems provides critical information to water quality scientists and engineers, particularly when the monitoring techniques can provide some information about the chemical nature of DOM. The efficacy of various indices derived from rapid, low-cost spectroscopic and chromatographic techniques to discriminate DOM composition was tested for samples prepared from well-defined mixtures of purified Aldrich humic acid (PAHA) and Suwannee River fulvic acid (SRFA). Sensitivities of the discrimination indices were examined by comparing (1) the differences between measured values and those predicted based from mass balance and the end member characteristics, and (2) the linear correlations between index values and mass ratios of the DOM mixtures. Size exclusion chromatography (SEC) results revealed that the weight average molecular weight (MW(w)) may be a useful approach for tracking DOM mixing processes, although the number-average molecular weight (MW(n)) may be better for distinguishing different DOM compositions. Specific ultraviolet absorbance measured at 254 nm (SUVA(254)) performed better as a discrimination index than did two previously recommended absorbance ratios, both in terms of making better predictions of intermediate compositions and in exhibiting a more linear correlation with PAHA mass ratio. Several well-defined peaks in the derivative absorption spectra (301 and 314 nm for the first derivative, 217 nm for the third derivative, and 211 and 224 nm for the fourth derivative) also were found to be promising potential DOM discrimination indices. Finally, a fluorescence ratio based on humic- versus fulvic-like fluorescence proved to be a superior DOM discrimination index for the two DOM end members studied here. In general, this study illustrates the evaluation process that should be followed to develop rapid, low-cost discrimination indices to monitor DOM compositions based on end member mixing analyses. PMID- 16307784 TI - Large-scale evaluation of the current level of polybrominated diphenyl ethers (PBDEs) in breast milk from 13 regions of Japan. AB - Polybrominated diphenyl ethers (PBDEs) were measured in 2004 in 105 breast milk samples collected from 13 regions of Japan (Hokkaido, Akita, Miyagi, Tokyo, Gifu, Fukui, Kyoto, Hyogo, Wakayama, Shimane, Yamaguchi, Kochi and Okinawa). Six congeners (BDE-28, BDE-47, BDE-99, BDE-100, BDE-153 and BDE-154) were determined by gas chromatography /mass spectrometry (GC/MS). Total PBDE levels ranged from 0.01 to 23.0 ng/g lipid (geometric mean (GM), 1.34 ng/g lipid). BDE-47 (GM, 0.66 ng/g lipid, 59% of sigmaPBDE) was the most abundant congener present in breast milk and was detected in 99% of the samples. Total PBDE levels were higher in northern Japan than in other regions. We analyzed the effects of occupation, age, smoking status, alcohol consumption and number of deliveries on total PBDE levels. None of these factors were significantly associated with the level of PBDEs. The present study revealed that the current level of exposure to PBDEs in Japan is lower than that in the USA or Sweden. GMs (ng/g lipid) (GSD, geometric standard deviation) and medians (ng/g lipid) of PBDE levels in each district are as follows: Hokkaido 2.70 (1.70), 2.74; Akita 4.49 (2.19), 5.44; Miyagi 1.77 (4.37), 1.11; Tokyo 1.39 (2.09), 1.63, Gifu 2.83 (4.79), 2.23; Fukui 1.05 (2.34), 1.18; Kyoto 1.31 (2.95), 1.33; Hyogo 1.02 (2.69), 0.88; Wakayama 1.33 (3.80), 1.70; Shimane 0.83 (2.51), 0.66; Yamaguchi 1.74 (2.82), 1.76; Kochi 0.50 (2.69), 0.74 and Okinawa 1.91 (2.75), 1.22. This is the first large-scale study of current PBDE levels in breast milk in Japan. PMID- 16307785 TI - QSPR study on soil sorption coefficient for persistent organic pollutants. AB - Quantitative structure-property relationship (QSPR) models of soil sorption coefficients for 32 persistent organic pollutants were constructed using our recently introduced Lu index and novel distance-based atom-type DAI topological indices. Using multiple linear regression technique, a 6-variable model was obtained with the correlation coefficient of estimations (R) being 0.95, and the standard error of estimations (s) being 0.23, and the correlation coefficient (R(cv)) and the standard error (s(cv)) in the leave-4-out cross-validation procedure are 0.90 and 0.31, respectively. The results in this study indicate that soil sorption coefficients of POPs are dominated by molecular size while some DAI indices have smaller influence. PMID- 16307786 TI - Occurrence and photochemical degradation of 17alpha-ethinylestradiol in Acushnet River Estuary. AB - 17alpha-Ethinylestradiol (EE2), a major constituent of common contraceptive pills, and three other estrogenic hormones, estrone (E1), 17beta-estradiol (E2) and mestranol (MeEE2) have been determined in Acushnet River Estuary seawater using a GC-MS technique. Among three estrogenic compounds detected, EE2 has the highest concentration, up to 4.7 ng/l, at which EE2 may affect lobster and other fish abundance in the coastal seawater due to its high biological activity on fish feminization. Two natural estrogenic hormones, E1 and E2 have also been found in the estuary at concentrations up to 1.2 ng/l and 0.83 ng/l, respectively. Although EE2 is persistent to microbial degradation, it can undergo a rapid photodegradation in estuarine seawater under natural sunlight irradiation, with a half-life of less than 1.5 days in spring sunny days. PMID- 16307787 TI - Separation of plastic mixtures using liquid-fluidized bed technology. AB - Separation of heavier-than-water plastic mixtures had been investigated via the fluidization of their packed beds induced by an upward flow of water. The samples examined were resin pellets and crushed plastic products including PVC, PET and PBT. On the onset of a flow, a mixed bed was swelled to the state of fluidization and separated into layers of respective resins depending on their density. The effects of the flow rate, an amount of the samples and their density difference were examined on the separation of resin pellets. Under an appropriate condition, the process was completed within a few minutes, and satisfactory separation was attained when the density difference of the samples exceeded 0.05 g cm-3. By using a column equipped with several sample outlets sorted resins could be recovered with their purity intact by withdrawing each layer successively from above through a suitable outlet. Multi-stage separation was also found to be effective in treating close-density samples. Compared with uniform-sized resin pellets, size distribution of crushed plastic samples deteriorated the separation to some extent. PMID- 16307788 TI - Quantitative structure-activity relationship models for prediction of sensory irritants (logRD50) of volatile organic chemicals. AB - Quantitative classification and regression models for prediction of sensory irritants (logRD50) of volatile organic chemicals (VOCs) have been developed. Each compound was represented by the calculated structural descriptors to encode constitutional, topological, geometrical, electrostatic, and quantum-chemical features. The heuristic method (HM) was then used to search the descriptor space and select the descriptors responsible for activity. The best classification results were found using support vector machine (SVM): the accuracy for training, test and overall data set is 96.5%, 85.7% and 94.4%, respectively. The nonlinear regression models were built by radial basis function neural networks (RNFNN) and SVM, respectively. The root mean squared errors (RMS) in prediction for the training, test and overall data set are 0.4755, 0.6322 and 0.5009 for reactive group, 0.2430, 0.4798 and 0.3064 for nonreactive group by RBFNN. The comparative results obtained by SVM are 0.4415, 0.7430 and 0.5140 for reactive group, 0.3920, 0.4520 and 0.4050 for nonreactive group, respectively. This paper proposes an effective method for poisonous chemicals screening and considering. PMID- 16307789 TI - Sequential removal of heavy metals ions and organic pollutants using an algal bacterial consortium. AB - The residual algal-bacterial biomass from photosynthetically supported, organic pollutant biodegradation processes, in enclosed photobioreactors, was tested for its ability to accumulate Cu(II), Ni(II), Cd(II), and Zn(II). Salicylate was chosen as a model contaminant. The algal-bacterial biomass combined the high adsorption capacity of microalgae with the low cost of the residual biomass, which makes it an attractive biosorbent for environmental applications. Cu(II) was preferentially taken-up from the medium when the metals were present both separately and in combination. There was no observed competition for adsorption sites, which suggested that Cu(II), Ni(II), Cd(II), and Zn(II) bind to different sites and that active Ni(II), Cd(II) and Zn(II) binding groups were present at very low concentrations. Therefore, special focus was given to Cu(II) biosorption. Cu(II) biosorption by the algal-bacterial biomass was characterized by an initial fast cell surface adsorption followed by a slower metabolically driven uptake. pH, Cu(II), and algal-bacterial concentration significantly affected the biosorption capacity for Cu(II). Maximum Cu(II) adsorption capacities of 8.5+/-0.4 mg g-1 were achieved at an initial Cu(II) concentration of 20 mg l-1 and at pH 5 for the tested algal-bacterial biomass. These are consistent with values reported for other microbial sorbents under similar conditions. The desorption of Cu(II) from saturated biomass was feasible by elution with a 0.0125 M HCl solution. Simultaneous Cu(II) and salicylate removal in a continuous stirred tank photobioreactor was not feasible due to the high toxicity of Cu(II) towards the microbial culture. The introduction of an adsorption column, packed with the algal-bacterial biomass, prior to the photobioreactor reduced Cu(II) concentration, thereby allowing the subsequent salicylate biodegradation in the photobioreactor. PMID- 16307790 TI - Oxalate and root exudates enhance the desorption of p,p'-DDT from soils. AB - The abiotic desorption of p,p'-DDT from seven Chinese soils spiked with p,p'-DDT and the effects of oxalate at 0.001-0.1M and the root exudates of maize, wheat, and ryegrass were evaluated using batch experiments. Soil organic carbon played a predominant role in the retention of DDT. Oxalate significantly increased the desorption of p,p'-DDT, with the largest increments ranging from 11% to 54% for different soils. Oxalate addition also resulted in the increased release of dissolved organic carbon and inorganic ions from soils. Root exudates had similar effects to those of oxalate. Root exudates significantly increased DDT desorption from the soils, and the general trend was similar among the plant species studied for all the soils (p > 0.05). Low molecular weight dissolved organic carbon amendments caused partial dissolution of the soil structure, such as the organo mineral linkages, resulting in the release of organic carbon and metal ions and thus the subsequent enhanced desorption of DDT from the soils. The enhancing effects of oxalate and root exudates on DDT desorption were influenced by the contents of soil organic carbon and dissolved organic carbon in soils. PMID- 16307791 TI - Health risk assessment on human exposed to environmental polycyclic aromatic hydrocarbons pollution sources. AB - To assess how the human exposure to environmental carcinogenic polycyclic aromatic hydrocarbons (PAHs) pollution sources generated from industrial, traffic and rural settings, we present a probabilistic risk model, appraised with reported empirical data. A probabilistic risk assessment framework is integrated with the potency equivalence factors (PEFs), age group-specific occupancy probability and the incremental lifetime cancer risk (ILCR) approaches to quantitatively estimate the exposure risk for three age groups of adults, children, and infants. The benzo[a]pyrene equivalents based PAH concentrations in rural, traffic, and industrial areas associated with age group-specific occupancy probability at different environmental settings are used to calculate daily exposure level through inhalation and dermal contact pathways. Risk analysis indicates that the inhalation-ILCR and dermal contact-ILCR values for adults follow a lognormal distribution with geometric mean 1.04x10(-4) and 3.85x10(-5) and geometric standard deviation 2.10 and 2.75, respectively, indicating high potential cancer risk; whereas for the infants the risk values are less than 10( 6), indicating no significant cancer risk. Sensitivity analysis indicates that the input variables of cancer slope factor and daily inhalation exposure level have the greater impact than that of body weight on the inhalation-ILCR; whereas for the dermal-ILCR, particle-bound PAH-to-skin adherence factor and daily dermal exposure level have the significant influence than that of body weight. PMID- 16307792 TI - Thin endometrial echo complex on ultrasound does not reliably exclude type 2 endometrial cancers. AB - OBJECTIVE: The objective of this study was to determine the ultrasonographic characteristics of the uterus and endometrial echo-complex (EEC) of postmenopausal patients diagnosed with type 2 endometrial cancer, including uterine papillary serous carcinoma (UPSC), clear cell carcinoma (CCC), and other endometrial high-grade carcinomas (HGC). METHODS: Postmenopausal patients with type 2 endometrial cancer who underwent preoperative pelvic ultrasound were identified. Histologic diagnoses were made by biopsy or hysterectomy. Ultrasound reports were abstracted for the following parameters: EEC thickness, presence of intracavitary fluid or lesion, myometrial mass, uterine size, and adnexal mass. Ultrasound films were re-reviewed by a single gynecologic ultrasonographer. RESULTS: Fifty-two patients with detailed ultrasound reports comprised the study population. Twenty-six films were available for re-review and inter-examiner agreement was 92%. Forty-four women (85%) presented with abnormal vaginal bleeding. Thirty-four patients (65%) had a thickened EEC measuring >5 mm. In 9 cases (17%), the EEC measured <4 mm. In an additional 9 patients (17%) the EEC was indistinct. One or more other ultrasound abnormalities were found in each of the 18 cases with a non-thickened EEC: intracavitary fluid or lesion (8; 44%), myometrial mass (12; 67%), enlarged uterus (13; 72%), or adnexal mass (5; 28%). CONCLUSION: A thin or indistinct endometrial stripe, especially when associated with other ultrasound abnormalities does not reliably exclude type 2 endometrial cancer. Postmenopausal vaginal bleeding requires endometrial sampling despite a thin EEC in the presence of other ultrasonographic abnormalities or persistent bleeding. PMID- 16307793 TI - Management of borderline ovarian tumors: results of an Italian multicenter study. AB - OBJECTIVE: A rising number of patients affected by ovarian masses suspected of borderline ovarian tumor are operated on primary laparoscopic procedure. METHODS: From January 1992 to June 2004, 113 patients affected by low malignant ovarian tumor were followed at the Gynecologic Departments of five Italian Institutions. Fifty-two (46.0%) patients were operated on laparoscopic surgery, whereas traditional laparotomic approach was preferred in 61 cases (54%). In 53 patients (46.9%), a fertility-sparing surgical treatment was chosen. RESULTS: The diameter of the ovarian cysts ranged between 20 and 300 mm; in 20 out 113 patients (17.7%), the borderline tumor was bilateral. In 22/113 (19.5%) cases, we observed tumor rupture or spilling during surgery, and this incidence was greater in the group of patients treated by laparoscopy compared to laparotomy, and this difference is statistically significative. In 13/113 (11.5%) patients, we observed a relapse; the incidence of relapse, however, is not dependent on the type of surgical approach. The progression-free survival is higher for stages IA IC if compared to stages more than Ist and the difference is statistically significant. Type of surgical approach, laparoscopic versus laparotomic, does not seem to influence the PFS; when cystectomy is performed, the PFS is significantly lower if compared with demolitive surgery or monolateral annessectomy. Eight pregnancies were obtained: in one case, miscarriage was observed, whereas 7 healthy babies were born all but two by vaginal delivery. CONCLUSIONS: Conservative laparoscopic surgery may be the treatment of choice; an attractive option is that this surgery should be performed by an oncologist surgeon to obtain correct surgical staging and better results in terms of both relapse-free survival and pregnancies. PMID- 16307794 TI - Paclitaxel-loaded poly(gamma-glutamic acid)-poly(lactide) nanoparticles as a targeted drug delivery system for the treatment of liver cancer. AB - The study was to develop paclitaxel-loaded formulations using a novel type of self-assembled nanoparticles (P/NPs) composed of block copolymers synthesized by poly(gamma-glutamic acid) and poly(lactide). For the potential of targeting liver cancer cells, galactosamine was conjugated on the prepared nanoparticles (Gal P/NPs). In the in vitro studies, it was found that both the P/NPs and the Gal P/NPs had a similar release profile of paclitaxel. The activity in inhibiting the growth of HepG2 cells by the Gal-P/NPs was comparable to that of a clinically available paclitaxel formulation (Phyxol), while the P/NPs displayed a significantly less activity (p<0.05). The biodistribution and anti-tumor efficacy of the prepared nanoparticles were studied in hepatoma-tumor-bearing nude mice. It was found that the groups injected with Phyxol, the P/NPs or the Gal-P/NPs significantly delayed the tumor growth as compared to the control group injected with PBS (p<0.05). Among all studied groups, the group injected with the Gal P/NPs appeared to have the most significant efficacy in the reduction of the size of the tumor. This is because a large number of the Gal-P/NPs were observed at the tumor site, and subsequently released their encapsulated paclitaxel to inhibit the growth of the tumor. The aforementioned results indicated that the Gal-P/NPs prepared in the study had a specific interaction with the hepatoma tumor induced in nude mice via ligand-receptor recognition. Therefore, the prepared Gal-P/NPs may be used as a potential drug delivery system for the targeted delivery to liver cancers. PMID- 16307795 TI - Rapid localized cell trapping on biodegradable polymers using cell surface derivatization and microfluidic networking. AB - Spatial control over cell attachment is essential for controlling cell behavior and engineering cell-based sensor arrays. Here we report on a patterning procedure that can be utilized on a wide range of adherent and non-adherent cell types without the need to identify the exact peptide sequence or extracellular matrix (ECM) necessary for optimal cell attachment. This is achieved by converting native sialic residues present on the surface of most cells into non native aldehydes using a mild sodium periodate treatment. The aldehyde groups are then reacted with biotin hydrazide to produce biotinylated cells. Avidin is patterned onto the surface of a biotinylated biodegradable block copolymer, polylactide-poly(ethylene glycol)-biotin (PLA-PEG-biotin) by microfluidic networking using a PDMS stamp. The biotinylated cells then bind specifically to the patterned avidin regions. The PEG that is presented from the PLA-PEG-biotin copolymer in the regions without avidin immobilization minimizes cell binding in the non-patterned regions. PMID- 16307796 TI - Bone formation in a long bone defect model using a platelet-rich plasma-loaded collagen scaffold. AB - Platelet-rich plasma (PRP), a platelet concentrate made of autogenous blood, has been used in recent years to improve bone defect healing particularly in maxillofacial reconstructions. The aim of the present study was to assess the effect of PRP on new bone formation in a critical diaphyseal long bone defect. A critical size defect (2.5 cm) in the tibial diaphysis of 16 sheep was supplied either with autogenous PRP in a collagen carrier or with collagen alone (controls). Platelets were enriched about 3.5 fold compared to normal blood in the PRP. After 12 weeks, the explanted bone specimens were quantitatively assessed by X-ray, computed tomography (CT), biomechanical testing and histological evaluation. Bone volume, mineral density, mechanical rigidity and histology of the newly formed bone in the defect did not differ significantly between the PRP treated and the control group, and no effect of PRP upon bone formation was observed. It was suggested that PRP does not enhance new bone formation in a critical size defect with a low regenerative potential. Such bone defects might require more potent stimuli, e.g. combinations of functional biomaterials or autografts, precursor cells or osteoinductive growth factors. PMID- 16307797 TI - Development of a large titanium bone chamber to study in vivo bone ingrowth. AB - In the bone conduction chamber (BCC) various materials and factors have been tested for their effect on bone graft incorporation and bone healing. However, biomaterials often have to be crushed to fit in this small chamber. Since cellular responses to biomaterials are influenced by the size and shape of particles, research concerning the evaluation of biomaterials is limited by the dimensions of this bone chamber. We enlarged and modified the BCC in order to be able to investigate the in vivo influences of biomaterials, growth factors and bone graft processing on tissue and bone ingrowth. Seven goats received four bone chambers each, three modified models and a BCC. The first model (BCC+) had two ingrowth openings, similar to that of the BCC. The second model had two round ingrowth openings (ROU). The third model had a open bottom for bone ingrowth (BOT). After 12 weeks, bone ingrowth distances were measured on histological sections and using muCT. Bone ingrowth was significantly higher (p=0.009 and 0.008) in the ROU compared to the BCC+ and the BOT, respectively. Similar results were found using muCT. The ROU model performed most similar to the BCC (gold standard) and is considered to be a promising new tool in biomaterials research. PMID- 16307798 TI - Continuous monitoring of WBC (biochemistry) in an adult leukemia patient using advanced FTIR-spectroscopy. AB - Fourier transform infrared (FTIR)-spectroscopy has been found useful for monitoring the effectiveness of drugs during chemotherapy in leukemia patients. In the present work, spectral changes that occurred in the white blood cells (WBC) of an adult acute myeloid leukemia (AML) patient and their possible utilization for monitoring biochemistry of WBC were investigated. The phosphate absorbance from nucleic acids and the lipid-protein ratio in the WBC decreased immediately after treatment and then increased to levels of a control group. Similar observations were recorded in child patients with acute lymphoblastic leukemia (ALL) who were used as test cases. These parameters maybe used as possible markers to indicate successful remission and suggest that FTIR spectroscopy may provide a rapid optical method for continuous monitoring or evaluation of a WBC population. PMID- 16307799 TI - Lactobacillus endocarditis: case report and review of cases reported since 1992. AB - Lactobacillus is a rare cause of endocarditis with only 78 reported cases in the literature. We report a case of Lactobacillus endocarditis and review reported adult cases since the introduction of the Duke's criteria for endocarditis. An analysis is presented with a focus on outcomes. Mortality from reported cases of Lactobacillus endocarditis was found to have substantially improved since 1992. Surgical rates for persistent disease remain the same. Possible reasons for this improvement include better antibiotics, earlier recognition of cases, and more aggressive treatment regimens. PMID- 16307800 TI - The influence of tonsillitis on oral and throat muscles in children. AB - OBJECTIVES: Surface electromyography (sEMG) studies were performed on 80 children with acute tonsillitis (AT) and 110 children with recurrent tonsillitis (RT), age 4-12 years, to trace sEMG changes of duration and amplitude of muscle activity during swallowing and continuous drinking. STUDY DESIGN: Prospective observational study of randomly chosen patients compared to normative database. METHODS: Timing and amplitude (in muV) of activity of masseter, submental and infrahyoid muscles were evaluated during voluntary single swallows of saliva ("dry" swallow), water swallows, swallows of excessive amount of water (up to 15 ml), and continuous drinking of 50 ml of water. These parameters were measured for two age groups for both conditions: 4-8 and 9-12 years old. The previously established normative database was taken for control. RESULTS: AT presents prolonged duration of swallowing and electric hyperactivity of infrahyoid muscles but this activity returns to normal after recovery. RT affects masseter and infrahyoid muscles even during periods of remission but do not affect duration of swallowing activity. CONCLUSION: AT and RT in children age 4-12 years affects muscle activity during swallowing significantly by involving additional muscles (mainly infrahyoid) in this process. AT presents temporary electric hyperactivity of infrahyoid muscles. RT affects masseter and infrahyoid muscles even during periods of remission (pathologic changes are fixed). Abnormally high electric activity of masseter and infrahyoid muscles in patients with RT might serve as an additional indicator for tonsillectomy. Surface EMG of swallowing is a simple, non-invasive and reliable method for diagnostic and preoperative evaluation of dysphagia complaints associated with tonsillitis. PMID- 16307801 TI - A novel small reporter gene and HIV-1 fitness assay. AB - Most currently available HIV-1 reporter gene constructs are large and disrupt the nef reading frame. This report describes a novel reporter gene based on the small murine heat stable antigen (HSA) protein, which is expressed on the surface of infected cells. This HSA reporter can be inserted in the vpr reading frame, leaving nef intact. Nine amino acids from the extracellular domain of HSA are replaced with an influenza hemagglutinin (HA) antibody epitope (HSA-HA). Like the parental reporter protein, this novel reporter is expressed on the surface of infected cells. Antibodies for HSA and HA specifically detect reporter viruses with each construct, indicating disruption of the original HSA antibody epitope. Finally, a strategy is developed to detect each reporter virus by real-time PCR quantitation. The growth of viruses tagged with each reporter allows precise assessment of the relative growth of viruses differing in mutations of interest. Moreover, the availability of these reporters in either of two half-genome plasmids allows convenient production of reporter and non-reporter HIV-1 by co transfection of appropriately paired plasmids. These paired reporter viruses offer a potentially useful standardized method for measurement of HIV-1 fitness in competition assays. PMID- 16307802 TI - A simple wax-embedding method for isolation of aphid hemolymph for detection of luteoviruses in the hemocoel. AB - A protocol for isolating hemolymph from viruliferous aphids has been developed. This method uses warm melted wax to immobilize the aphid. Following removal of a hind leg, the hemolymph can be collected readily. Flushing with RNase-free water allows for collection of sufficient hemolymph for RNA extraction from individual aphids. The extracted RNA was successfully used for detection of barley yellow dwarf virus (BYDV) and pea enation mosaic virus (PEMV) from individual viruliferous Rhopalosiphum padi and Acyrthosiphon pisum aphids, respectively. A TaqMan real-time RT-PCR protocol for quantitation of PEMV in the hemolymph of individual aphids was developed. The wax-embedding hemolymph collection technique provides a useful tool for studying molecular interactions between persistent and circulative plant viruses and their insect vectors. PMID- 16307803 TI - Unique organisation of tRNA genes in Entamoeba histolytica. AB - The genome sequence of the protistan parasite Entamoeba histolytica HM-1:IMSS has been completed recently. Among the findings has been a unique organisation for the tRNA genes in this organism. Forty-two of the tRNA isoacceptor types are encoded in tandem arrays that vary in unit length from 490 to 1775 basepairs and contain from 1 to 5 tRNA genes. In three cases a 5S RNA gene is also present in the unit. An estimated 10% of the genome is made up of these arrays. Interspersed between RNA-encoding sequences are short tandem repeats that are polymorphic between isolates and, in some cases, within isolates. The number and organisation of tRNA genes in E. histolytica is unprecedented. In addition to encoding the tRNAs of the organism we propose that the arrays may fulfil a structural role in the genome. PMID- 16307804 TI - T-DNA tagging and characterization of a cryptic root-specific promoter in Arabidopsis. AB - From a T-DNA tagged Arabidopsis population, a line, M-57 showing GUS (beta glucuronidase) expression in the vascular regions of young roots was identified. Southern analysis revealed presence of a single T-DNA insert. Using inverse PCR, the plant sequence flanking the T-DNA insertion was cloned. The insertion was identified to be in the intergenic area between loci At4G13940 and At4G13930, coding for SAHH (S-Adenosyl-l-Homocysteine Hydrolase) and SHMT (Serine Hydroxy Methyl Transferase) genes, respectively. A 452-bp fragment immediately upstream of the T-DNA insertion when cloned and mobilized as a GUS fusion was capable of driving a similar root-specific expression of reporter gene in transgenic Arabidopsis plants and their progenies. This cryptic promoter element does not show the presence of any known root-specific promoter element. PMID- 16307805 TI - Simple criteria for differentiation of Fabry disease from amyloid heart disease and other causes of left ventricular hypertrophy. AB - AIMS: Fabry disease may be difficult to differentiate from other causes of left ventricular hypertrophy such as other myocardial storage diseases (including amyloidosis), hypertrophic cardiomyopathy (HCM), or hypertensive heart disease (HHD). We sought to determine simple criteria to best differentiate the above mentioned cardiac diseases. METHODS AND RESULTS: All patients in a six-year time period with left ventricular hypertrophy due to Fabry disease (13 patients), biopsy proven cardiac amyloidosis (16 patients), non-obstructive HCM (17 patients), and 22 randomly selected patients with advanced HHD were compared. Retrospective analysis of clinical characteristics, findings of electrocardiogram (ECG) and echocardiography by blind review was performed. RESULTS: No single clinical characteristic or findings of ECG or echocardiography could reliably differentiate between the various diseases. Increased echogenicity/granular sparkling, valvular abnormalities, abnormal renal function, and diastolic function were not helpful discriminators. In a univariate analysis, four criteria (acroparesthesia, anhydrosis, absence of hypertension and presence of Sokolow criteria for left ventricular hypertrophy in the ECG) were significant for Fabry disease. By logistic regression analysis, the following most suitable discriminative parameters were identified: hypertension in HHD (specificity 82%), orthostasis and/or pericardial effusion for amyloidosis (specificity 93%), papillary muscle anomaly in non-obstructive HCM (specificity 92%), and Fabry disease if neither hypertension orthostatis, pericardial effusion nor a papillary muscle anomaly was present (specificity 87%). CONCLUSION: A combination of symptoms, echocardiographic findings and ECG in unexplained left ventricular hypertrophy may help to differentiate amyloidosis, non-obstructive HCM and hypertensive heart disease from Fabry disease. The results of this preliminary study will have to be confirmed in a prospective study. PMID- 16307806 TI - Increasing post-event plasma thrombomodulin level associates with worse outcome in survival of acute coronary syndrome. AB - OBJECTIVES: We seek to evaluate the association of serial plasma soluble thrombomodulin (TM) change and outcome in survivals of acute coronary syndrome (ACS). BACKGROUND: Plasma TM is a marker of endothelial damage. Elevated cross sectional TM level has been found to be associated with worse outcome in survivals of acute myocardial infarction (MI). However, the association of serial TM change with ACS is not clear. METHODS: In 47 survivals of ACS [39 males; age, 64.7+/-9.3 years; 15 unstable angina (UA), 32 acute MI], plasma soluble TM and serum high sensitivity C-reactive protein (hs-CRP) levels are measured at index ACS and at 3-month follow-up. Using hierarchical cluster analysis, patients are classified into ascending (n=29) and descending group (n=18) according to the change in TM level. Composite study end-point is recurrent UA, MI and sudden cardiac death (SCD). RESULTS: The magnitude of change of plasma soluble TM is not related to demographic characteristics, diagnosis and pattern of revascularization. The change of TM level in ascending and descending group is 1.80+/-1.13 and -1.73+/-0.48 ng/ml (p<0.001), respectively. There is no significantly difference in demographic characteristics, diagnosis and pattern of revascularization between both groups. The mean hs-CRP levels at index ACS and at 3-months follow-up are also not statistically different between two groups. At 6 month follow-up, 6 patients in ascending group reach end-point (1 SCD, 1 UA, and 4 MIs), however, no patient in descending group suffers from subsequent event (p=0.044). After a mean follow-up of 317 days (range, 60 to 541 days), 9 (19%) patients reach end-point. Kaplan-Meier survival analysis reveals that increasing plasma TM predicts worse clinical outcome (hazard ratio=0.13, log-transformed 95% confidence interval 0.03 to 0.48, p=0.0224 by the log-rank test). CONCLUSION: Increasing plasma TM level associates with worse outcome in patients surviving ACS. PMID- 16307807 TI - Comparison of myocardial contrast echocardiography with SPECT in the evaluation of coronary artery disease in asymptomatic patients with LBBB. AB - OBJECTIVE: The non-invasive assessment of coronary artery disease (CAD) in patients with left bundle branch block (LBBB) is troublesome. In this study, we investigated the diagnostic accuracy of myocardial contrast echocardiography (MCE) with adenosine to detect CAD in asymptomatic patients with LBBB, and we compared it with single photon emission computed tomography (SPECT) with adenosine. METHODS: Forty-seven patients with LBBB, and no previously documented CAD, initially underwent SPECT imaging and 1-3 days later MCE. Coronary arteriography was performed within 1 week from the latter procedure. RESULTS: The overall sensitivity, specificity, positive predictive value, negative predictive value, diagnostic accuracy, and kappa index of concordance of SPECT were 73%, 72%, 44%, 90%, 72%, and 0.37+/-0.13, respectively, whereas those of MCE were 91%, 92%, 77%, 97%, 92%, and 0.77+/-0.1, respectively (p<0.05 for all comparisons). Significant CAD was present in 11 patients (23%). Left anterior descending coronary artery was involved in 8 patients, left circumflex artery in 2 patients, and right coronary artery in 4 patients. Concerning the left anterior descending artery disease detection, SPECT had a sensitivity of 75%, a specificity of 79%, a positive predictive value of 43%, a negative predictive value of 94%, and a diagnostic accuracy of 79%. The respective values of MCE were 100% for all of the above variables. CONCLUSIONS: MCE with adenosine has a higher global diagnostic accuracy compared to SPECT for the detection of CAD in patients with LBBB, mainly due to the poor specificity of SPECT concerning perfusion defects detection in the left anterior descending artery territory. PMID- 16307808 TI - Dengue myocarditis, rare but not fatal manifestation. PMID- 16307809 TI - Transcriptional activity of genes encoding Transforming Growth Factor beta and its receptors in peripheral blood mononuclear cells from patients with acute coronary syndromes. AB - BACKGROUND: Recent data report altered gene expression of numerous pro- and anti inflammatory factors involved in pathology of acute coronary syndromes (ACS). Transforming growth factor beta (TGFbeta) signaling is engaged in a wide range of processes. Its effect on vessels seems to be protective due to its anti inflammatory and anti-atherogenic action. However, it also seems to be engaged in such negative effects as neointima formation and fibrosis. The aim of the study was to assess the expression of the genes encoding TGFbeta and its receptors (type I, II, and III) in patients with ACS. METHODS: The study was carried out on 24 patients with acute coronary syndrome (7 with unstable angina [UA] and 17 with myocardial infarction [MI]) and 10 age-matched healthy subjects (control). To evaluate gene expression of TGFbeta and its receptors total mRNA was extracted from peripheral blood mononuclear cells (PBMC) and the number of mRNA copies were assessed by quantitative reverse transcriptase polymerase chain reaction (QRT PCR). RESULTS: MI and UA patients demonstrated significantly lower TGFbeta gene expression compared to control (2789+/-418 c/microg vs. 20262+/-2548 c/microg; p<0.001, and 3390+/-518 c/microg vs. 20262+/-2548 c/microg; p<0.001, respectively), as well as noticeably lower transcriptional activity of genes encoding its type I (3295+/-447 c/microg vs. 12859+/-1929 c/microg; p<0.001, and 3258+/-721 c/microg vs. 12859+/-1929 c/microg; p<0.01, respectively) and type II receptors (2364+/-346 c/microg vs. 19003+/-2357 c/microg; p<0.001, and 2680+/-522 c/microg vs. 19003+/-2357 c/microg; p<0.01, respectively). Also, gene expression of the type III receptor was inferior in the studied group compared to the control, although the difference was significant only for the UA group vs. control. Expressions of the studied genes did not differ between patients with MI and those with UA. CONCLUSION: Our report shows that the decreased activity of TGFbeta in patients with ACS is at least partly due altered transcriptional activity of genes encoding both TGFbeta and its receptors, what may be responsible for the evolution of atherosclerotic lesions. PMID- 16307810 TI - Comparison of clinical and angiographic features and longterm follow-up events between patients with variant angina and patients with ST elevation myocardial infarction. AB - We investigated to what extent patients with variant angina and significant coronary stenosis (>or=70%) present a clinical and angiographic profile similar to patients with ST elevation myocardial infarction. Thus, the clinical and angiographic features as well as follow-up events of 200 patients were prospectively analyzed and were compared with those of 422 patients with a first ST elevation myocardial infarction survivors of the early phase (3 days) and those of 70 patients with variant angina and non significant stenosis. Age and incidence of smoking, systemic hypertension, diabetes and maximum ST elevation were similar in the 2 groups. Furthermore, among patients with significant coronary stenosis, stenosis severity and the proportion of eccentric lesions were also comparable. Incidence of recent-within 30 days prior to admission-angina at rest was higher in variant angina patients with significant stenosis (67% vs. 27%, p<0.001) than in those with myocardial infarction but long standing angina at rest (>30 days) was low and comparable in these 2 groups (15% vs. 11%, ns). Also, in a 5-year follow-up most patients from these 2 groups were free from angina at rest (86% vs. 84%) which in variant angina patients was largely attributable to a high revascularization rate (72%). Moreover, the rate of myocardial infarction/cardiac death (20% vs. 19%) was also similar. Patients with variant angina and non-significant stenosis, however, had longer antecedent angina, more frequent follow-up angina and a lower incidence of cardiac events than the other 2 groups. Thus, these findings suggest that patients with variant angina and significant coronary stenosis generally behave as an acute coronary syndrome-likely associated with an acutely complicated plaque-rather than as recurrent vasospastic angina, and should be managed accordingly. PMID- 16307811 TI - Polymer conjugates: nanosized medicines for treating cancer. AB - Interdisciplinary research at the interface of polymer chemistry and the biomedical sciences has produced the first polymer-based nanomedicines for the diagnosis and treatment of cancer. These water-soluble hybrid constructs, designed for intravenous administration, fall into two main categories: polymer protein conjugates or polymer-drug conjugates. Polymer conjugation to proteins reduces immunogenicity, prolongs plasma half-life and enhances protein stability. Polymer-drug conjugation promotes tumor targeting through the enhanced permeability and retention (EPR) effect and, at the cellular level following endocytic capture, allows lysosomotropic drug delivery. The successful clinical application of polymer-protein conjugates (PEGylated enzymes and cytokines) and promising results arising from clinical trials with polymer-bound chemotherapy (e.g. doxorubicin, paclitaxel, camptothecins) has provided a firm foundation for more sophisticated second-generation constructs that deliver the newly emerging target-directed anticancer agents (e.g. modulators of the cell cycle, signal transduction inhibitors and antiangiogenic drugs) in addition to polymer-drug combinations (e.g. endocrine- and chemo-therapy). PMID- 16307812 TI - The curative role of radiotherapy in adenocarcinoma of the prostate in patients under 55 years of age: a rare cancer network retrospective study. AB - To determine whether radiation therapy could be an acceptable alternative to surgery in young patients with adenocarcinoma of the prostate, we analysed the outcome of 39 patients aged under 55 with organ confined tumours who received external radiation therapy in a curative intent. Our results suggest that similar local control in younger and older patients can be expected from either external beam radiotherapy or radical prostatectomy. PMID- 16307813 TI - Preoperative hyperfractionated accelerated radiotherapy and radical surgery in advanced head and neck cancer: a prospective phase II study. AB - BACKGROUND AND PURPOSE: To evaluate whether preoperative hyperfractionated accelerated radiotherapy (RT) combined with major radical surgery is feasible and successful in the treatment of advanced primary head and neck cancer. PATIENTS AND METHODS: Ninety four patients with histologically confirmed head and neck squamous cell cancer (HNSCC) in the oral cavity (41/96; 43%), supraglottis (14/96; 15%), glottis (5/96; 5%), oropharynx (16/96; 17%), nasal cavity/paranasal sinuses (8/96; 8%), nasopharynx (3/96; 3%), hypopharynx (7/96; 7%) and two (2%) with unknown primary tumour and large cervical lymph nodes entered into the study. 21/96 patients (22%) had stage II, 17/96 (18%) stage III and 58/96 patients (60%) stage IV disease. The patients received preoperative hyperfractionated RT 1.6 Gy twice a day, 5 days a week to a median tumour dose of 63 Gy with a planned break for 11 days (median) after the median dose of 37 Gy. Then, after a median of 27 days the patients underwent major radical surgery of the primary tumour and metastatic lymph nodes including reconstructions with pedicled or microvascular free flaps when indicated as a part of the scheduled therapy. 12/96 patients had only ipsilateral or bilateral neck dissections. RESULTS: After a median follow-up time of 37.2 mos 77/96 (80.2%) patients had complete locoregional control. All but 2 patients had complete histological remission after surgery. 40/96 pts were alive without disease, two of them after salvage surgery. 32/96 patients had relapsed; 15 had locoregional and 13 distant relapses, 4 patients relapsed both locoregionally and distantly. Fifty patients have died; 29 with locoregional and/or distant relapse, eight patients died of second malignancy, and 19 had intercurrent diseases. Disease-specific and overall survival at 3 years was 67.7 and 51%, respectively. Acute grade three mucosal reactions were common, but transient and tolerable. Late grade 3-4 adverse effects were few. CONCLUSIONS: Preoperative hyperfractionated accelerated RT can be successfully combined with major radical surgery in the treatment of HNSCC. The amount of serious late adverse effects was not increased. PMID- 16307815 TI - Concomitant boost radiotherapy with concurrent weekly cisplatin in advanced head and neck cancers: a phase II trial. PMID- 16307816 TI - Detection and frequency of VT1, VT2 and eaeA genes in Escherichia coli O157 and O157:H7 strains isolated from cattle, cattle carcasses and abattoir environment in Istanbul. AB - The aim of this study was to detect VT1, VT2 and eaeA genes and to determine the frequency of these genes in Escherichia coli O157 and O157:H7 strains isolated from cattle, cattle carcasses and environmental samples of the 5 abattoirs located in Istanbul, Turkey. For this, the presence of VT1, VT2 and eaeA genes in 26 strains of E. coli O157:H7 and 6 strains of O157 was investigated by multiplex PCR. The results have shown that eaeA gene was detected in all O157 and O157:H7 strains tested. Both VT2 and eaeA genes were detected in 4 (80%) of 5 strains of E. coli O157 and eaeA alone in 1 strain of O157. In 27 strains of O157:H7, 5 (18.5%) strains were found to be positive for VT1, VT2 and eaeA genes, 19 (70.3%) strains for both VT2 and eaeA and, 3 (11.1%) strains for only eaeA gene. Either VT1 alone or VT2 alone was not detected in any strains tested. eaeA gene alone in 2 strains, VT2-eaeA genes in 9 strains and VT1-VT2-eaeA genes in 2 strains were detected in 13 of E. coli O157:H7 strains isolated from cattle. eaeA alone in 1 strain, VT2-eaeA genes in 5 strains and VT1-VT2-eaeA genes in 2 strains were detected in 8 of E. coli O157:H7 strains isolated from carcasses. VT2-eaeA genes in 5 strains (isolated from hands, apron, knife and floor) and VT1-VT2-eaeA genes in 1 strain (isolated from knife) were also detected in 6 of E. coli O157:H7 strains isolated from environmental samples. This study reveals that most of the strains are found to be toxigenic and it is most likely that strains isolated from carcasses and abattoir environment originated from cattle feces. Therefore, HACCP systems are necessary from farm to table especially in the abattoirs to prevent contamination of meat and abattoir environment with intestinal content. PMID- 16307817 TI - End-to-end communication in the modulation of translation by mammalian RNA viruses. AB - A 5'-3' end interaction leading to stimulation of translation has been described for many cellular and viral mRNAs. Enhancement of viral translational efficiency mediated by 5' and 3' untranslated regions (UTRs) has been shown to occur via RNA RNA interactions or novel RNA-protein interactions. Mammalian RNA viruses make use of end-to-end communication in conjunction with both viral and cellular factors to regulate multiple processes including translation initiation and the switch between translation and RNA synthesis during the viral lifecycle. PMID- 16307818 TI - Functional interactions between the Epstein-Barr virus BZLF1 protein and the promyelocytic leukemia protein. AB - The Epstein-Barr virus immediate-early protein BZLF1 (Z) has been shown to alter the cellular localization of the promyelocytic leukemia (PML) protein. PML has important implications for growth control, apoptosis, anti-viral effects and many more processes. Here we further examined the relationship between PML and the Epstein-Barr virus Z protein. We examined the effect of Z expression on PML protein levels, and the effect of increased PML protein levels on Z-mediated dispersion of PML bodies. We found that increased levels of PML protein, such as through interferon treatment, were able to suppress Z-mediated PML body dispersion. We also studied the consequences of PML dispersion by Z, by examining p21 transactivation, A20 transactivation, and MHC Class I presentation levels in Z-expressing cells. We found that, while Z-mediated dispersion of PML did not affect MHC Class I presentation, it did alter p21 and A20 expression. In addition, we found that increased levels of PML were able to prevent Z protein binding to mitotic chromosomes. Our work implies that the balance of PML and Z levels in cells may affect how each protein functions. PMID- 16307819 TI - Synonymous codon usage in adenoviruses: influence of mutation, selection and protein hydropathy. AB - Trends in synonymous codon usage in adenoviruses have been examined through the multivariate statistical analysis on the annotated protein-coding regions of 22 adenoviral species, for which complete genome sequences are available. One of the major determinants of such trends is the G+C content at third codon positions of the genes, the average value of which varied from one viral genome to other depending on the overall mutational bias of the species. G3S and C3S interacted synergistically along the first principal axis of correspondence analysis on the Relative Synonymous Codon Usage of adenoviral genes, but antagonistically along the second principal axis. The intra-genomic variation in codon usage pattern in adenoviruses is generally influenced by asymmetrical mutational bias in two DNA strands. Other major determinants of the trends are the natural selection, putatively operative at the level of translation and quite interestingly, hydropathy of the encoded proteins. The trends in codon usage, though characterized by distinct virus-specific mutational bias, do not exhibit any sign of host-specificity. Significant variations are observed in synonymous codon choice in structural and nonstructural genes of adenoviruses. PMID- 16307820 TI - Divergent IRES elements in invertebrates. AB - Viruses have evolved unique strategies and mechanisms to recruit ribosomes to ensure continued translation of their viral RNA during infection. The Dicistroviridae family of invertebrate viruses contains an unusual internal ribosome entry site (IRES), which can directly recruit ribosomes in the absence of initiation factors. Moreover, this IRES initiates translation at a non-AUG codon independent of an initiator Met-tRNA. Recent studies have shown that the IRES mimicks a tRNA to interact with and manipulate the ribosome. The presence of this divergent IRES likely allows translation of the dicistroviral RNA during infection when host translation is compromised. This review will explore the unique properties of this unprecedented mechanism of gene expression. Specific topics will examine structural components of the IRES, the mechanism of initiating translation at non-AUG codons and the regulation of this IRES in vivo. The existence of this mechanism suggests that the repertoire of open reading frames in our genome may be greater than anticipated. PMID- 16307821 TI - Cloning and characterization of differentially expressed genes of internal breakdown in mango fruit (Mangifera indica). AB - Internal breakdown in mango fruits has become a major concern in recent years. This disorder renders the fruits unfit for human consumption. The overall loss due to this disorder is about 35-55%. Environmental and physiological factors like high temperature, humidity, respiration and low transpiration rates have been attributed to cause spongy tissue due to reduced loss of heat from fruits. Biochemical studies have shown that there is a reduction in pH, total soluble solids, ascorbic acid, total sugars and carotenoids, low reducing and non reducing sugar contents, lower amylase and invertase activities and high acid and starch content in spongy tissue affected pulp. There are no reports on molecular studies to determine changes in gene expression in these tissues. The present study was conducted using PCR based subtractive hybridization and RNA gel blot analysis of a few selected genes. The latter showed a higher expression of catalase, ubiquitin, alcohol dehydrogenase, coproporphyrinogen oxidase and keratin associated protein. A lower expression of ribosomal gene, fructose bisphosphate aldolase and cysthathionine gamma synthase was also noticed in spongy tissue. Biochemical studies indicated a lower amylase activity and a lower content of the total and reducing sugars in spongy tissue as compared to healthy tissue. Analyses of results indicate that oxidative stress may be one of the causes for formation of spongy tissue, which affects the expression of many genes. The role of these genes in spongy tissue formation is discussed. PMID- 16307822 TI - Rate, risk factors and outcomes of catheter-related bloodstream infection in a paediatric intensive care unit in Saudi Arabia. AB - The aim of this study was to determine the rate, risk factors and outcomes of catheter-related bloodstream infections (CRBSIs) in patients in a paediatric intensive care unit (PICU). A prospective cohort study was performed in King Abdulaziz Medical City, Riyadh, Saudi Arabia; a 650-bed academic/tertiary care centre with a combined 10-bed medical and surgical PICU. All patients admitted to the PICU from July 2000 to February 2003 who had a central line placed were monitored for the development of bloodstream infection (BSI) from insertion until 48 h after removal. Four hundred and forty-six patients with 2493 central-line days were documented; 273 (55%) were male and the mean age was 2.6 years. Of the 446 patients, 278 (56%) had congenital heart disease, 108 (22%) had genetic disorders and/or congenital malformations, 55 (11%) had respiratory disease, and 42 (8%) had trauma. There were 50 episodes of CRBSI in 46 patients with a rate of 20.06 per 1,000 central-line-days and a device-utilization rate of 57%. Of these 50 episodes, 24 (48%) were polymicrobial, 16 (32%) were due to Gram-negative organisms, five (10%) were due to Gram-positive organisms, and five (10%) were fungal. The most common organisms isolated were Klebsiella pneumoniae (N=12, 16%), coagulase-negative staphylococci (N=10, 14%) and Pseudomonas aeruginosa (N=8, 11%). The mean duration of line insertion was 11.8 days for CRBSI patients and 4.22 days for non-BSI patients (P<0.0001). The mean PICU stay was 30.20 days for CRBSI patients and 6.35 days for non-BSI patients (P<0.0001). BSI occurred more often in catheters inserted in the PICU compared with the operating room, and in the femoral site compared with jugular or subclavian sites (P<0.001). In multiple logistic regression analysis of the risk factors, CRBSI patients were more likely to have multiple central lines [odds ratio (OR) 9.19; 95% confidence intervals (CI): 3.76-22.43), the line was more likely to be used for total parenteral nutrition (OR: 8.69; 95% CI: 3.5-21.4), and guidewire exchange was more likely to be performed on the line. CRBSI was not associated with a higher mortality rate. The CRBSI rate in our hospital is high compared with that reported by the National Nosocomial Infection Surveillance system. This study has established a benchmark for future comparisons. Additional studies from Saudi Arabia are necessary for national comparison and development of preventive measures. PMID- 16307823 TI - Surveillance of healthcare-associated infections in Indonesian hospitals. AB - A cross-sectional surveillance of healthcare-associated infections (HAIs) and exposure to risk factors was undertaken in two Indonesian teaching hospitals (Hospitals A and B). Patients from internal medicine, surgery, obstetrics and gynaecology, paediatrics, a class department and intensive care were included. Patient demographics, antibiotic use, culture results, presence of HAI [phlebitis, surgical site infection (SSI), urinary tract infection (UTI) and septicaemia] and risk factors were recorded. To check for interobserver variation, a validation study was performed in Hospital B. In Hospitals A and B, 1,334 and 888 patients were included, respectively. Exposure to invasive devices and surgery was 59%. In Hospital A, 2.8% of all patients had phlebitis, 1.7% had SSI, 0.9% had UTI and 0.8% had septicaemia. In Hospital B, 3.8% had phlebitis, 1.8% had SSI, 1.1% had UTI and 0.8% had septicaemia. In the validation study, the prevalence as recorded by the first team was 2.6% phlebitis, 1.8% SSI, 0.9% UTI and no septicaemia, and that recorded by the second team was 2.2% phlebitis, 2.6% SSI, 3.5% UTI and 0.9% septicaemia. This study is the first to report on HAI in Indonesia. Prevalence rates are comparable to those in other countries. The reliability of the surveillance was insufficient as a considerable difference in prevalence rates was found in the validation study. The surveillance method used is a feasible tool for hospitals in countries with limited healthcare resources to estimate their level of HAI and make improvements in infection control. Efficiency can be improved by restricting the surveillance to include only those patients with invasive procedures. This can help to detect 90% of all infections while screening only 60% of patients. PMID- 16307824 TI - Efficacy of antiseptic-impregnated catheters on catheter colonization and catheter-related bloodstream infections in patients in an intensive care unit. AB - This study was conducted to evaluate the impact of central venous catheters impregnated with chlorhexidine and silver sulphadiazine on the incidence of colonization and catheter-related bloodstream infection in critically ill patients. One hundred and thirty-three patients requiring central venous catheterization were chosen at random to receive either an antiseptic-impregnated triple-lumen catheter (N=64) or a standard triple-lumen catheter (N=69). The mean (SD) durations of catheterization for the antiseptic and standard catheters were 11.7 (5.8) days (median 10; range 3-29) and 8.9 (4.6) days (median 8.0; range 3 20), respectively (P=0.006). Fourteen (21.9%) of the antiseptic catheters and 14 (20.3%) of the standard catheters had been colonized at the time of removal (P=0.834). Four cases (6.3%) of catheter-related bloodstream infection were associated with antiseptic catheters and one case (1.4%) was associated with a standard catheter (P=0.195). The catheter colonization rates were 18.7/1000 catheter-days for the antiseptic catheter group and 22.6/1000 catheter-days for the standard catheter group (P=0.640). The catheter-related bloodstream infection rates were 5.3/1000 catheter-days for the antiseptic catheter group and 1.6/1000 catheter-days for the standard catheter group (P=0.452). In conclusion, our results indicate that the use of antiseptic-impregnated central venous catheters has no effect on the incidence of either catheter colonization or catheter related bloodstream infection in critically ill patients. PMID- 16307825 TI - Prospective hospital-wide studies of 505 patients with nosocomial bacteraemia in 1997 and 2002. AB - We conducted surveys in 1997 and 2002 to determine the rate, underlying sources and outcome of nosocomial bacteraemia. Blood culture results were reviewed daily. All patients with positive cultures drawn >or=48 h after hospitalization were included in the study and their charts were reviewed. The underlying source of infection was determined by pre-defined clinical and/or microbiological criteria. Patients were followed until discharge or death. In 1997 and 2002, 851 and 857 patient-unique cases of bacteraemia were diagnosed, respectively, excluding contaminants; of these, 228 (27%) and 277 (32%) cases, respectively, were hospital acquired (P<0.05). The overall rate decreased from 7.5 to 7.0 per 1,000 admissions (P<0.001). The sources of bacteraemia in 1997 and 2002, respectively, were: intravascular catheters (36% and 27%, P<0.05), urinary tract (8% and 15%, P<0.05), respiratory tract (5% and 13%, P<0.01) and surgical sites (14% and 4%, P<0.001). In one-third of patients, the source of bacteraemia could not be determined. Only 52% and 54%, respectively, of these patients were discharged alive (difference was not significant). In 1997, Staphylococcus aureus was the most frequent isolate (26%), followed by coagulase-negative Staphylococcus (13%) and Klebsiella pneumoniae (11%). By 2002, the incidence of S. aureus had fallen to 11% (P<0.001), acinetobacter was the single most frequently isolated organism (increased from 6% to 17%) (P<0.001). In-hospital mortality associated with acinetobacter bacteraemia (57%) was significantly higher than that for other organisms (31-43%) (P<0.05). In conclusion, prospective surveys of nosocomial bacteraemias provide valuable information, facilitating the pursuit of successful interventions. PMID- 16307826 TI - Virucidal activity of a new hand disinfectant with reduced ethanol content: comparison with other alcohol-based formulations. AB - A new formula with reduced ethanol content (55%) in combination with 10% propan-1 ol, 5.9% propan-1.2-diol, 5.7% butan-1.3-diol and 0.7% phosphoric acid exhibited a broad spectrum of virucidal activity. In quantitative suspension tests, with and without protein load, this formulation reduced the infectivity titre of seven enveloped (influenza A and B, herpes simplex 1 and 2, bovine corona, respiratory syncytial, vaccinia, hepatitis B, bovine viral diarrhoea) and four non-enveloped (hepatitis A, polio, rota, feline calici) viruses >10(3)-fold within 30s. In comparative testing, only 95% ethanol showed similar levels of activity. In fingerpad tests, the formulation produced a log10 reduction factor of the titre of poliovirus type 1 (Sabin) of 3.04 in 30s compared with 1.32 by 60% propan-2 ol. Testing against feline calicivirus produced a log10 reduction factor of 2.38 by the test formulation; in contrast, the log10 reduction factors with 70% ethanol and 70% propan-1-ol were 0.68 and 0.70, respectively. PMID- 16307827 TI - Evidence from peptidomic analysis of skin secretions that the red-legged frogs, Rana aurora draytonii and Rana aurora aurora, are distinct species. AB - The northern red-legged frog Rana aurora aurora and the California red-legged frog Rana aurora draytonii are traditionally classified together in the same species group. Ten peptides with antimicrobial activity were isolated from norepinephrine-stimulated skin secretions of R. aurora draytonii and purified to near homogeneity. The peptides were identified as belonging to the ranatuerin-2 family (two peptides), brevinin-1 family (four peptides), temporin family (three peptides), and a novel peptide, RV-23 (RIGVLLARLPKLFSLFKLMGKKV) that has limited structural similarity to the bee venom peptide, melittin. This distribution of peptides contrasts with that found previously in skin secretions from R. aurora aurora collected under the same conditions and at the same time of year (one ranatuerin-2 peptide, two brevinin-1 peptides, and one temporin peptide). The variation in amino acid sequences between corresponding R. aurora draytonii and R. aurora aurora peptides is comparable with the variation in sequences of orthologs from other members of the Amerana group of New World ranid frogs (Rana boylii, Rana muscosa, and Rana luteiventris). It is proposed, therefore, that the red-legged frogs should be regarded as separate species (R. aurora and R. draytonii) within the Amerana group rather than conspecific subspecies. The data emphasize that amino acid sequences of antimicrobial peptides in skin secretions may be used to infer taxonomic and phylogenetic relationships between species of ranid frogs. PMID- 16307829 TI - Serum MCP-1 levels are increased in mild cognitive impairment and mild Alzheimer's disease. AB - Upregulation of a number of chemokines, including monocyte chemotactic protein-1 (MCP-1), is associated with Alzheimer's disease (AD) pathological changes. Emerging evidence suggests that inflammatory events precede the clinical development of AD, as cytokine disregulation has been observed also in patients with mild cognitive impairment (MCI). MCP-1 levels were evaluated in serum samples from 48 subjects with MCI, 94 AD patients and 24 age-matched controls. Significantly increased MCP-1 levels were found in MCI and mild AD, but not in severe AD patients as compared with controls. mRNA levels in peripheral blood mononuclear cells (PBMC), evaluated by quantitative RT-PCR analysis, paralleled serum MCP-1 levels. Moreover, a progressive MCP-1 decrease was observed over a 1 year follow up in a subgroup of MCI subjects converted to AD. MCP-1 upregulation is likely to be a very early event in AD pathogenesis, by far preceding the clinical onset of the disease. Nevertheless, as MCP-1 is likely to play a role in several pathologies with an inflammatory component, a possible usefulness as an early AD biomarker would be possible only in combination with other molecules. PMID- 16307830 TI - Early (in uterus and infant) exposure to mercury and lead. AB - Mercury and lead are toxic metals widely spread in the environment with bio accumulative features that raises public health concerns. Both metals are equally dispersed in the human food chain but exposure and risk of toxicity during early human development are modulated by the diet and nutritional status. Understanding how Hg and Pb occur and interact with nutrients is fundamental to establish guidelines for diminishing exposure and the risk of toxicity. The risk of fetal and infant exposure to Hg can be influenced by maternal amalgam filling (inorganic Hg) and fish consumption (monomethyl Hg), whereas the risk of exposure to Pb is complex: maternal absorption depends on nutrient interactions (Ca and P); and maternal body Pb accumulation responds to all factors known to interact with bone and calcium metabolism. Maternal exposure to Hg and Pb is more important during fetal development than during breastfeeding. Moreover, these metals (especially Pb) are frequently higher in infant formulas which do not carry the nutritional and psychological advantages and protection of breastfeeding. Infant's reference dose is lower for Hg than for Pb, but risk of Pb contamination for fetuses and infant (breast- or formula-fed) is higher and lasts longer than Hg. Breastfeeding is essential to complete infant development. Interruption or suppression of breast-feeding with cow's milk-based formulas is not an option to environmental pollution. PMID- 16307831 TI - Validation of the clinical approach of using the induced serum iron increase after 1h as a measure of iron absorption. AB - BACKGROUND & AIMS: To evaluate the correlation between the iron absorption and the induced increase in serum iron concentration (S-Fe) after 1 h. METHODS: Eleven subjects were after an over night fast served a wheat roll fortified with 100-mg iron as ferrous sulfate labelled with radio iron. The S-Fe response and the iron absorption measured by whole body counting, were studied. RESULTS: There was a significant correlation between the iron absorption and the S-Fe increase (R2 = 0.75, P < 0.001). CONCLUSION: This short and valid observation period proposes the S-Fe procedure as an uncomplicated and clinically applicable tool when evaluating the iron absorption capacity of patients. PMID- 16307832 TI - Low-dose intraperitoneal Freund's adjuvant: toxicity and immunogenicity in mice using an immunogen targeting amyloid-beta peptide. AB - Complete Freund's adjuvant (CFA) is effective for potentiating immune responses in mice when administered subcutaneously, and is often more potent when given intraperitoneally (i.p.). However, the the potential toxicity of i.p. administration in mice has led investigators and Institutional Animal Care and Use committees to increasingly view the use of CFA i.p. with reservation. We evaluated whether an 80% reduction in the dose of CFA administered i.p. to mice, compared to the i.p. doses used in a previous analysis, could abrogate the untoward effects associated with its use, while still maintaining adjuvanticity. Using a novel immunogen targeting the N-terminus of the 42-amino acid amyloid beta peptide, we compared low dose CFA administered i.p., with three other commonly used adjuvants given i.p.: alum, incomplete Freunds adjuvant (IFA) and monophoshoryl lipid A + trehalose dicorynomycolate (MPL + TDM). The results of the study showed that, though the reduction in intraperitoneal dose of CFA mitigated transient weight loss and leukocytosis observed previously with higher doses of i.p. CFA, all mice administered CFA or IFA i.p. developed abdominal adhesions and granulomatous peritonitis. Mice from all adjuvant groups, however, appeared to tolerate the respective adjuvants well and excellent comparative immunogenicity was observed in mice immunized with the Freunds and MPL + TDM adjuvants. Consequently, we conclude that though a high-titered, humoral response may be generated using low dose CFA administered i.p., the accompanying toxicity remains significant, and thus alternative adjuvants and/or routes should be considered. PMID- 16307833 TI - Development of an opsonin inhibition assay for evaluation of complex polysaccharide protective epitopes. AB - The induction of opsonic antibodies directed against capsular polysaccharides (Ps) is an important mechanism by which immunization protects against the development of invasive pneumococcal (Pn) infection. In preparing Pn vaccines, it is necessary to compare different manufacturing lots of capsular Ps, or to compare oligosaccharides used for conjugate vaccines with native capsular Ps, in order to insure that important epitopes of the Ps are maintained. We have developed an opsonic-antibody inhibition assay (OIA) to compare the functional epitopes of different capsular Ps preparations in vitro. Components of the OIA are primary neutrophils, rabbit complement (C'), and type-specific antibody (Ab). After conditions for optimal opsonic killing were determined for each Pn serotype, anti-Pn Ab was pre-incubated with different dilutions of purified capsular Ps, then added to the OIA mix. Plotting the % bacteria killed versus Ps concentration (log transformed) yielded a linear curve that was used to quantify the concentration of capsular Ps which inhibited the bacteria killing by 50% (IC50). The IC50 was determined for 8 Pn Ps types. These ranged between 6 ng/ml for type 6B and 1268 ng/ml for type 23F. Importantly OIA curves were statistically identical for two different manufacturing lots of capsular Ps for the 8 Pn Ps types. We conclude that differences among capsular Ps used for Pn vaccines could be detected with an OIA assay and these differences may predict the ability of Ps preparations to induce functionally active antibody when formulated into vaccines. PMID- 16307834 TI - Hepatitis A virus infection in the United States: serologic results from the Third National Health and Nutrition Examination Survey. AB - To determine the prevalence of hepatitis A virus (HAV) infection in the general U.S. population, sera from participants in the Third National Health and Nutrition Examination Survey (NHANES III) conducted in 1988-1994 were tested for antibody to HAV (anti-HAV). Among 21,260 participants aged > or = 6 years tested, the overall prevalence of infection was 31.3%, and increased markedly with age. The age-adjusted prevalence was significantly higher among foreign- compared to U.S.-born participants, and was highest among Mexican-Americans and lowest among non-Hispanic whites. Among U.S.-born children, only Mexican-American ethnicity and income below the poverty level were associated with HAV infection in a multivariate model. During this period before hepatitis A vaccination, age, ethnicity and birthplace were the most important determinants of HAV infection in the United States. PMID- 16307835 TI - Modern regulatory impact analysis: the experience of the European Union. AB - This paper begins by explaining the importance of improving the quality of the regulatory process, particularly in the European Union (EU), using tools and techniques such as Regulatory Impact Assessment (RIA). It describes the principal features of the RIA process, including good practices and practical problems. The paper then outlines the historical development of the RIA process in the EU and describes the new approach adopted recently by the European Commission. It also evaluates the effectiveness of the new approach--in theory and in practice--using as one example the RIAs undertaken on the New Chemicals Policy (REACH) by the European Commission, Member States, and stakeholders. A further section describes recent work undertaken by Business Decisions Limited and THE WEINBERG GROUP on Impact Assessments for the Animal Health Industry, the Polycarbonates Industry and the Nickel Industry. The paper finally draws a number of conclusions and makes a number of recommendations for further improvements in the quality of the regulatory process in the European Union. PMID- 16307836 TI - Husbands' and wives' reports of women's decision-making power in Western Guatemala and their effects on preventive health behaviors. AB - Surveys have attempted to measure married women's decision-making power by asking women who has a say and/or final say in a number of household decisions. In several studies where the same questions were posed to husbands, considerable discrepancies in reports were found. This paper assesses husband and wife reports of decision-making on four matters (whether or not to buy household items; what to do if a child becomes ill; whether or not to buy medicine for a family member who is ill; what to do if a pregnant women becomes very ill) and the relationship of these reports to three recent health behaviors (having an emergency plan during pregnancy; delivering in a health facility; having a postpartum checkup within 4 weeks). A sample of 1000 women in 53 communities in three departments of western Guatemala was selected using a stratified random sampling approach. A standard household questionnaire was used to identify the respondents as well as to obtain data on household characteristics. Husbands of interviewed women were interviewed in every other household giving information on 546 couples for this analysis. Women and men's questionnaires were similar and were designed to obtain information on the respondent's knowledge, attitudes and behaviors regarding maternal health. Consistent with other research, results show that relative to their husbands' report, wives tend to under-report their household decision making power. In couples with both partners educated and in couples in which women work for pay, both partners were significantly more likely to report that both of them participate in the final decisions than was the case in couples without education or in which the wife did not work for pay. Women's reports of their decision-making power was significantly related to the household having a plan for what to do in case of a maternal emergency, but was not associated with place of childbirth or with having a postpartum checkup, while husband's reports of the wife's decision-making power was negatively associated with the likelihood of having the last birth in a health facility. PMID- 16307837 TI - Low-dose venlafaxine in three adolescents and young adults with autistic disorder improves self-injurious behavior and attention deficit/hyperactivity disorders (ADHD)-like symptoms. AB - In our clinical practice, we have had good experiences with venlafaxine in the treatment of self-injurious behavior (SIB) and attention deficit/hyperactivity disorders (ADHD)-like symptoms in patients with pervasive developmental disorders (PDD), and we report here three cases of possible therapeutic response: (A) a 17 year-old boy with autism and severe behavioral symptoms, including aggression toward self or property, SIB and hyperactivity, who appeared to respond to low dose venlafaxine (18.75 mg/day); (B) a 23-year-old woman with autism hyperactivity who appeared to respond to low-dose venlafaxine (18.75 mg/day); (C) a 17-year-old girl with autism hyperactivity who appeared to respond to low-dose venlafaxine (18.75 mg/day). Follow-ups occurred respectively 18, 36 and 6 months after treatment initiation, making it possible to observe the stability of the clinical improvement in these cases. PMID- 16307838 TI - T-2 toxin induces apoptosis, and selenium partly blocks, T-2 toxin induced apoptosis in chondrocytes through modulation of the Bax/Bcl-2 ratio. AB - T-2 toxin is one of the mycotoxins, a group of type A trichothecenes produced by several fungal genera including Fusarium species. In the present study, we have investigated the apoptotic effects of T-2 toxin on chondrocytes and the relationship between T-2 toxin induced chondrocyte apoptosis and its influence on Bcl-2/Bax protein and mRNA expression. We have also examined the inhibitory effects of selenium on chondrocyte apoptosis induced by T-2 toxin. We have combined morphological and biological techniques to establish the relevance of apoptosis in human chondrocyte death induced by T-2 toxin. Treatment with T-2 toxin caused accelerated apoptosis in a concentration dependent manner. The apoptosis induced by T-2 toxin involved an increased Bax/Bcl-2 ratio. Bcl-2 mRNA expression remained unchanged in chondrocyte apoptosis induced by T-2 toxin treatment, while Bax mRNA expression increased following treatment with T-2 toxin. Selenium could partly block the apoptosis of chondrocytes induced by T-2 toxin through decreasing the Bax/Bcl-2 ratio. These results suggest that, under our experimental conditions, apoptosis of chondrocytes can be induced by T-2 toxin (1-20ng/mL) via the Bcl-2 and Bax proteins, and the Bax/Bcl-2 ratio may play a critical role in governing the susceptibility to apoptosis induced by T-2 toxin in human chondrocytes. PMID- 16307839 TI - Chalcone inhibits the proliferation of human breast cancer cell by blocking cell cycle progression and inducing apoptosis. AB - Chalcones are discussed to represent cancer preventive food components in a human diet that is rich in fruits and vegetables. In this study, we examined chalcone (1,3-diphenyl-2-propenone) for its effect on proliferation in human breast cancer cell lines, MCF-7 and MDA-MB-231. The results showed that chalcone inhibited the proliferation of MCF-7 and MDA-MB-231 by inducing apoptosis and blocking cell cycle progression in the G2/M phase. Immunoblot assay showed that chalcone significantly decreased the expression of cyclin B1, cyclin A and Cdc2 protein, as well as increased the expression of p21 and p27 in a p53-independent manner, contributing to cell cycle arrest. An enhancement in Fas/APO-1 and its two form ligands, membrane-bound Fas ligand (mFasL) and soluble Fas ligand (sFasL), was responsible for the apoptotic effect induced by chalcone. In addition, chalcone also triggered the mitochondrial apoptotic signaling by increasing the amount of Bax and Bak and reducing the level of Bcl-2 and Bcl-X(L), and subsequently activated caspase-9 in MCF-7 and MDA-MB-231 cells. Taken together, our study suggests that the blockade of cell cycle progression and initiation of cell apoptotic system may participate in the antiproliferative activity of chalcone in human breast cancer cells. PMID- 16307840 TI - Cellular and molecular mechanisms of epilepsy in the human brain. AB - Animal models have provided invaluable data for identifying the pathogenesis of epileptic disorders. Clearly, the relevance of these experimental findings would be strengthened by the demonstration that similar fundamental mechanisms are at work in the human epileptic brain. Epilepsy surgery has indeed opened the possibility to directly study the functional properties of human brain tissue in vitro, and to analyze the mechanisms underlying seizures and epileptogenesis. Here, we summarize the findings obtained over the last 40 years from electrophysiological, histochemical and molecular experiments made with the human brain tissue. In particular, this review will focus on (i) the synaptic and non synaptic properties of neocortical neurons along with their ability to produce synchronous activity; (ii) the anatomical and functional alterations that characterize limbic structures in patients presenting with mesial temporal lobe epilepsy; (iii) the issue of antiepileptic drug action and resistance; and (iv) the pathophysiology of seizure genesis in Taylor's type focal cortical dysplasia. Finally, we will address some of the problems that are inherent to this type of experimental approach, in particular the lack of proper controls and possible strategies to obviate this limitation. PMID- 16307841 TI - Cost analysis of alternative methods for wastewater handling in small communities. AB - Wastewater collection and treatment is quite important for sustainable management. It would be uneconomical and impractical to provide sewer systems and separate wastewater treatment plants (WWTP) for small communities. The decision process in wastewater planning is rather important in terms of comparing the alternatives considered. The two important points in the management of wastewater at rural areas not connected to a sewer system are to develop an optimized operation strategy and to make sure that the complete system is environmentally and economically sustainable. In some regions, package treatment could be an alternative solution. However, in cases where there is an existing large WWTP, a cluster system, where sewage generated by small communities could be transported via conveyors to a centralized WWTP, could be employed. In this study, the wastewater treatment and disposal problems in small communities were addressed and an alternative wastewater handling scenario was proposed. Additionally, three wastewater handling scenarios were compared. As a case study, Gebze villages were selected. PMID- 16307842 TI - Application of a system dynamics approach for assessment and mitigation of CO2 emissions from the cement industry. AB - A system dynamics model based on the dynamic interactions among a number of system components is developed to estimate CO(2) emissions from the cement industry in India. The CO(2) emissions are projected to reach 396.89 million tonnes by the year 2020 if the existing cement making technological options are followed. Policy options of population growth stabilisation, energy conservation and structural management in cement manufacturing processes are incorporated for developing the CO(2) mitigation scenarios. A 42% reduction in the CO(2) emissions can be achieved in the year 2020 based on an integrated mitigation scenario. Indirect CO(2) emissions from the transport of raw materials to the cement plants and finished product to market are also estimated. PMID- 16307843 TI - Effect of cystathionine beta-synthase variant 844ins68bp and methylenetetrahydrofolate reductase A1298C polymorphisms in xenografts on 5-FU efficacy and doubling time. AB - The association of MTHFR and CBS variants with the doubling time and responsiveness to several chemodrugs was analyzed in 26 human cancer xenografts. The tumors homozygous for the absence of insertion (NN) for the CBS 844ins68bp were more chemosensitive than those with insertion (NI) to TS-1 (P=0.0048), suggesting a potential effect of this variant on fluoropyrimidine efficacy. Furthermore, the doubling time of tumors with a variant C allele (AC or CC) in MTHFR-A1298C was significantly longer than that of tumors with a normal allele (AA) (P=0.0008). Twenty-nine cellular proliferation-related genes were associated with MTHFR-A1298C genotyping and with the doubling time. PMID- 16307844 TI - Screening of new sorbent materials for cadmium removal from aqueous solutions. AB - This study compares the abilities of four low-cost materials: peels of peas, broad bean, and medlar, respectively and fig leaves, to remove cadmium from aqueous solutions. Kinetic data and equilibrium sorption isotherms were measured in batch conditions. Kinetics of cadmium sorption was contact time, initial cadmium concentration and sorbent type dependent. The results also showed that the kinetics of cadmium sorption were described by a pseudo second-order rate model. The cadmium uptake of these low-cost materials was quantitatively evaluated using sorption isotherms. Results indicated that Langmuir model gave an acceptable fit to the experimental data. A high cadmium sorption was observed by these materials. The broad bean peel was the most effective to remove cadmium ions with a maximum sorption capacity about 147.71 mg/g followed by peas peel (118.91 mg/g), fig leaves (103.09 mg/g), and medlar peel (98.14 mg/g). PMID- 16307845 TI - Sex differences in how older students influence younger student smoking behaviour. AB - The purpose of this study was to examine sex differences in how the smoking behaviour of close friends and older peers at school influence smoking onset and progression among 10,843 grade 9, 10 and 11 students from 29 secondary schools in Ontario, Canada. In lower-risk student populations (i.e., students with no smoking friends or one smoking friend), males and females were both at a similar level of risk for occasional smoking as a function of the smoking rate of older students at their school. Among higher-risk student populations (i.e., students with three or more close friends who smoke), the smoking rate of older students at school did not influence the risk of occasional smoking. The odds of a female student being a regular smoker increased as the prevalence of smoking among older students at her school increased regardless of her close friends' smoking behaviour. However, among male students, the influence of older student smoking at school on the odds of regular smoking was moderated by the number of close friends who smoke. It appears that the prevalence of older student smoking at a school is more influential among younger female students, whereas the smoking behaviour of close friends appears to be more influential among younger male students. This new finding provides unique insight for tailoring and targeting future school-based smoking prevention initiatives. PMID- 16307846 TI - Motivating smoking reductions by framing health information as safer smoking tips. AB - Explored the hypothesis that framing health information as safer smoking tips might motivate change in cigarette smokers. Daily smokers were randomly assigned to a control condition or to an intervention condition in which respondents were asked if they knew about a range of different safer smoking tips. Respondents (43 daily smokers, 18 years or older, 58% male) who received safer smoking tips reported a small reduction in the number of cigarettes smoked (p=.05) by the 3 month follow-up as compared to respondents in the control condition. This pilot result is encouraging but requires replication before considering the value of including safer smoking tips in health messages for smokers in the general population. PMID- 16307847 TI - PKC-mediated modulation of L-type calcium channels may contribute to fat-induced insulin resistance. AB - Increased intracellular free calcium [Ca2+]i has been noted in adipocytes, platelets, and leukocytes of subjects with insulin resistance syndrome or allied disorders. In rodent studies, measures which increase [Ca2+]i in adipocytes and skeletal muscle are associated with impaired insulin signaling, attributable at least in part to diminished ability of insulin to activate phosphoserine phosphatase-1 (PP-1). In fat-fed insulin resistant rats, pre-treatment with a drug that selectively chelates intracellular calcium eliminates about half of the decrement in insulin-stimulated glucose uptake induced by fat feeding; since this chelator does not influence the insulin sensitivity of chow-fed rats, it is reasonable to suspect that fat feeding boosts [Ca2+]i in skeletal muscle, and that this effect is partially responsible for the associated reduction in insulin sensitivity. Clinical insulin resistance is associated with increased levels of triglycerides and other fatty acid metabolites in muscle fibers; this can give rise to diacylglycerol-mediated activation of PKC, which in turn compromises insulin signaling by triggering kinase cascades that phosphorylate IRS-1 on key serine residues. Yet there is also evidence that, in skeletal muscle, PKC activity up-regulates the function of L-type calcium channels, increasing their maximal conductance while left-shifting their voltage dependence. Thus, the PKC activation associated with fat overexposure might be expected to boost basal [Ca2+]i in skeletal muscle, potentially impeding insulin-mediated activation of PP-1. This hypothesis is consistent with several clinical studies demonstrating that long-acting inhibitors of L-type calcium channels can improve insulin sensitivity in overweight hypertensives; it should be readily testable in rodent models of fat-induced insulin resistance. Since parathyroid hormone can act on adipocytes and muscle to boost [Ca2+]i, mild secondary hyperparathyroidism associated with low calcium intakes and poor vitamin D status may contribute to insulin resistance, consistent with certain clinical and epidemiological findings. Magnesium, often thought of as a mild calcium antagonist, appears to have favorable effects on insulin sensitivity and risk for diabetes, and recent evidence indicates that increases of intracellular magnesium within the physiological range can diminish calcium influx through phosphorylated L-type calcium channels. It will be of interest to determine whether calcium antagonism does indeed underlie the favorable influence of good magnesium status on insulin function. A report that chromium picolinate can induce the plasmalemmal Ca2+ ATPase in smooth muscle cells, raises the possibility that modulation of calcium transport might play a role in the insulin-sensitizing efficacy of bioactive chromium. PMID- 16307848 TI - "The stone which the builders rejected...": Delay of reinforcement and response rate on fixed-interval and related schedules. AB - The article deals with response rates (mainly running and peak or terminal rates) on simple and on some mixed-FI schedules and explores the idea that these rates are determined by the average delay of reinforcement for responses occurring during the response periods that the schedules generate. The effects of reinforcement delay are assumed to be mediated by a hyperbolic delay of reinforcement gradient. The account predicts that (a) running rates on simple FI schedules should increase with increasing rate of reinforcement, in a manner close to that required by Herrnstein's equation, (b) improving temporal control during acquisition should be associated with increasing running rates, (c) two valued mixed-FI schedules with equiprobable components should produce complex results, with peak rates sometimes being higher on the longer component schedule, and (d) that effects of reinforcement probability on mixed-FI should affect the response rate at the time of the shorter component only. All these predictions were confirmed by data, although effects in some experiments remain outside the scope of the model. In general, delay of reinforcement as a determinant of response rate on FI and related schedules (rather than temporal control on such schedules) seems a useful starting point for a more thorough analysis of some neglected questions about performance on FI and related schedules. PMID- 16307849 TI - Mitochondrial phylogeny of Anura (Amphibia): a case study of congruent phylogenetic reconstruction using amino acid and nucleotide characters. AB - We explore whether phylogenetic analyses of the same sequence data set at the amino acid and nucleotide level are able to recover congruent topologies, as well as the advantages and limitations of both alternative approaches. As a case study, mitochondrial protein-coding genes were used to discern among competing hypotheses on the phylogenetic relationships of major anuran amphibian lineages. To properly address this phylogenetic question, the complete nucleotide sequences of the mitochondrial genomes of two archaeobatrachian species, Ascaphus truei and Pelobates cultripes, were determined anew. Bayesian and maximum likelihood phylogenetic inferences of the same sequence data set were performed based on both amino acid and nucleotide characters, with the latter analysed either as codons or as a reduced data set of first+second (P12) codon positions. In addition, likelihood-based ratio tests were performed to evaluate the support of alternative topologies. The different data sets arrived at congruent and highly supported topologies, suggesting a similar phylogenetic resolving power of the two character types provided that correctly selected sites and appropriate evolutionary models are used. The reconstructed anuran mitochondrial phylogeny supports the paraphyly of Archaeobatrachia, with Ascaphus as sister group to all the remaining anurans, and Pelobates as sister group of Neobatrachia. However, the employed tree reconstruction methods and likelihood-based ratio tests seemed to be negatively affected by the fast evolving sequences of neobatrachians, suggesting that the phylogeny of Anura here presented is not definitive, and needs further investigation using an extended taxon sampling. PMID- 16307850 TI - Identification of an epithelial-specific enhancer regulating ESX expression. AB - The Ets transcription factor, ESX, exhibits a unique pattern of epithelial restricted expression and transactivates genes involved in epithelial differentiation and cancer. The aim of this study was to determine the underlying genetic basis for epithelial-specific expression of ESX. We have identified a 30bp ESX enhancer sequence (EES) approximately 3 kb upstream of the proximal promoter. This region displays enhancer activity in an epithelial-specific manner and deletion of this region abrogates ESX gene transcription. An EES binding protein complex (EBC) was identified through electrophoretic mobility shift assays whose degree of EES binding correlated well with endogenous ESX levels in epithelial cells and was regulated by epithelial differentiation. Understanding the regulation of this element will lend insight into mechanisms of epithelial differentiation and the etiology of breast cancer and may provide novel targets for cancer therapeutic intervention. PMID- 16307851 TI - Interactions between dendritic cells and epithelial cells in allergic disease. AB - Dendritic cells (DCs) play a crucial role in the sensitisation process. Upon encounter with an allergen, DCs require interactions with other cells and factors for triggering a primary or secondary immune response. Epithelial cells (ECs) express features of accessory cells, such as expression of HLA-DR, co-stimulatory molecules, functional FcgammaR, molecules of the antigen-processing machinery, and display an ability to internalise antigen. These features may authorize them to function as immunomodulators (e.g. amplification of memory T cells during secondary immune responses). ECs may increase chemokine (e.g. CCL20) secretion thereby attracting DCs. Epithelial human TSLP activates DC, which allow them to prime naive T cells for the production of pro-inflammatory cytokines, while down regulating IFN-gamma and IL-10. ECs may also influence the local polarization of types l and 2 antigen-presenting cells via PGE(2) by impairing the ability of maturing DC to produce bioactive IL-12 p70. PGE(2) is synergistic with IL-1beta and TNF-alpha in the induction of functional and phenotypic maturation of DC and induce IL12 p40 production. Sensitisation via the respiratory route may be Th(2) skewed, possibly because the antigen recognition by DC occurs in an environment rich of airway EC-product such as PGE(2). PMID- 16307852 TI - Spontaneous activity in the inferior colliculus of CBA/J mice after manipulations that induce tinnitus. AB - Several physiological studies have linked experimentally induced tinnitus to increases in the spontaneous activity of auditory neurons. These results have led to the proposal of hyperactivity models of tinnitus in which elevated neural activity in the absence of auditory stimulation is perceived as phantom sound. Such models are appealing in their simplicity but remain controversial because a generalized elevation of spontaneous rates may not be observed after treatments that induce tinnitus in humans and experimental animals. Our study addressed these issues by characterizing the effects of common methods of tinnitus induction on spontaneous activity in the central nucleus of the inferior colliculus (ICC). The ICC is an interesting structure in tinnitus research because its diverse inputs include putative generator sites in the dorsal cochlear nucleus, as well as brainstem sources that appear to remain normal after tinnitus induction. Groups of CBA/J mice were subjected to one of three induction methods: bilateral or unilateral sound exposure, and acute salicylate intoxication. Relative to normal baselines, bilaterally exposed mice showed increases in the spontaneous rates of neurons with tuning near the exposure frequency. When the sample was separated into physiologically defined response classes, exposure effects were strongest among neurons with broad excitatory bandwidths. By contrast, salicylate decreased the spontaneous rates of low frequency neurons with transient sound-evoked activity. Our results suggest that the disordered processes of hearing that give rise to tinnitus do not involve a pervasive elevation of spontaneous activity or a single mode of induction. PMID- 16307853 TI - Mineralocorticoid receptor mediates glucocorticoid treatment effects in the autoimmune mouse ear. AB - The standard treatment for many hearing disorders is glucocorticoid therapy, although the cochlear mechanisms involved in steroid-responsive hearing loss are poorly understood. Cochlear dysfunction in autoimmune mice has recently been shown to be controlled with the mineralocorticoid aldosterone as effectively as with the glucocorticoid prednisolone. Because aldosterone regulates sodium, potassium, and other electrolyte homeostasis, this implied the restoration of hearing with the mineralocorticoid was due to its impact on cochlear ion transport, particularly in the stria vascularis. This also suggested glucocorticoids may be controlling hearing recovery in part through their binding to the mineralocorticoid receptor in addition to their glucocorticoid receptor mediated anti-inflammatory and immunosuppressive functions. Therefore, the aim of the present study was to better delineate the role of the mineralocorticoid receptor in steroid control of hearing in the autoimmune mouse. Spironolactone, a mineralocorticoid receptor antagonist, was administered to MRL/MpJ-Fas(lpr) autoimmune mice in combination with either aldosterone or prednisolone to compare their hearing and systemic disease with mice that received either steroid alone. ABR thresholds showed either aldosterone or prednisolone alone preserved hearing in the mice, but spironolactone prevented both steroids from maintaining normal cochlear function. This suggested both steroids are preserving hearing through the mineralocorticoid receptor within the ear to regulate endolymph homeostasis. The spironolactone treatment did not block normal glucocorticoid receptor mediated immune-suppression functions because mice receiving prednisolone, either with or without spironolactone, maintained normal body weights, hematocrits, and serum immune complexes. Thus, reducing systemic autoimmune disease was not sufficient to control hearing if mineralocorticoid receptor-mediated functions were blocked. It was concluded the inner ear mineralocorticoid receptor is a significant target of glucocorticoids and a factor that should be considered in therapeutic treatments for steroid-responsive hearing loss. PMID- 16307854 TI - Spontaneous otoacoustic emissions from free-standing stereovillar bundles of ten species of lizard with small papillae. AB - Spontaneous otoacoustic emissions (SOAE) were measured in 10 lizard species from the families Iguanidae, Agamidae and Anguidae. The typical feature of these papillae is that the hair cells in the higher-frequency papillar regions that produce SOAE are not covered by a tectorial structure. The number of hair cells in the species used here was between 58 and 292 per ear. SOAE could be measured from all species, but some of their characteristics varied with papillar anatomy. Thus very small papillae produced fewer and smaller SOAE than larger papillae. PMID- 16307855 TI - Anti-inflammatory activity of cacalol and cacalone sesquiterpenes isolated from Psacalium decompositum. AB - The hexane extract and two sesquiterpenic compounds, cacalol and cacalone, were isolated from the roots and rhizomes of Psacalium decompositum. Then, their anti inflammatory activity was evaluated in carrageenan-induced rat paw edema and 12-O tetradecanoylphorbol-13-acetate (TPA)-induced ear edema. Indomethacin was used as the anti-inflammatory agent of reference. In the rat paw model of inflammation, both the hexane extract and the sesquiterpenes isolated from Psacalium decompositum showed a clear dose-dependent inhibition of the carrageenan-induced edema (P < 0.05), with important differences among them during the temporal course of the inhibition. In the TPA-induced mouse ear edema all tested compounds showed anti-inflammatory activity in dose-dependent manner (P < 0.05). In both models, cacalone showed the most prominent anti-inflammatory activity. We conclude that some of the beneficial effects attributed to Psacalium decompositum in traditional medicine can be related with the anti-inflammatory activity of cacalol and cacalone. PMID- 16307856 TI - Antinociceptive effect from Ipomoea cairica extract. AB - Ipomoea cairica L. Sweet (Convolvulaceae) is used in Brazilian folk medicine for the treatment of rheumatism and inflammations. Ipomoea cairica ethanolic extract (100, 300, 1000 and 3000 mg/kg; per os) induced dose-dependent reduction of response in the formalin test inflammatory phase in mice. The same dose range did not modify neurogenic pain in formalin test, tail-flick reflex latency, carrageenan-induced paw edema, and Rota-Rod test motor performance. From the bio active fraction 3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid were obtained. These compounds have been previously reported to have analgesic and antioxidative effects. A possible explanation for the antinociception is that somehow the compounds present in the extract reduced the release of pro nociceptive mediators unrelated to carrageenan-induced edema, such as histamine. Interestingly, caffeoylquinic acid derivatives have been reported to inhibit histamine release on in vitro models. The isolated caffeoylquinic acids could explain, at least in part, the antinociceptive effect of Ipomoea cairica polar extract. PMID- 16307857 TI - Estimation of the mtDNA mutation rate in aging mice by proteome analysis and mathematical modeling. AB - The accumulation of mitochondria containing mutated genomes was proposed to be an important factor involved in aging. Although the level of mutated mtDNA has shown to increase over time, it is currently not possible to directly measure the mtDNA mutation rate within living cells. The combination of mathematical modeling and controlled experiments is an alternative approach to obtain an estimate for the mutation rate in a well-defined system. In order to judge the relevance of mitochondrial mutations for the aging process, we used a mouse model to study age related alterations of the mitochondrial proteins. Based on these experimental data we constructed a mathematical model of the mitochondrial population dynamics to estimate mtDNA mutation rates. Mitochondria were isolated from mouse brain and liver at six different ages (newborn to 24-months). A large-gel 2D electrophoresis-based proteomics approach was used to analyze the mitochondrial proteins. The expression of two respiratory chain complex I subunits and one complex IV subunit decreased significantly with age. One subunit of complex III and one subunit of complex V increased in expression during aging. Together, these data indicate that complex I and IV deficiency in aged tissues might be accompanied by feedback regulation of other protein complexes in the respiratory chain. When we fitted our experimental data to the mathematical model, mtDNA mutation rate was estimated to be 2.7x10(-8) per mtDNA per day for brain and 3.2x10(-9) per mtDNA per day for liver. According to our model and in agreement with the mitochondrial theory of aging, mtDNA mutations could cause the detrimental changes seen in mitochondrial populations during the normal lifespan of mice, while at the same time ensure that the mitochondrial population remains functional during the developmental and reproductive period of mice. PMID- 16307858 TI - Decreased plasma cholesterol levels during aging in transgenic mouse models of Alzheimer's disease. AB - A large number of studies deals with the association of cholesterol and Abeta levels, however, the results are so far controversial. Whereas some studies report on increased cholesterol levels, other authors refer to an association of decreased peripheral cholesterol and the incidence of Alzheimer's disease. It is also questionable whether plasma cholesterol levels could be used as a predictive biomarker for the incidence of Alzheimer's disease. In the present report, we studied the relationship between these two parameters during aging in different transgenic mouse models of Alzheimer's disease, expressing both mutant human amyloid precursor protein and mutant human presenilin-1. Measurements of plasma cholesterol levels revealed a significant reduction in aged APP/PS1 and APP/PS1ki mice, whereas plasma levels in young and aged control mice remained almost unchanged. Furthermore, statistical analysis revealed a significant negative correlation between plasma cholesterol and brain Abeta42 levels during aging in the mice expressing both APP and PS1. PMID- 16307859 TI - Sensitive and selective liquid chromatography-electrospray ionisation-mass spectrometry analysis of astragaloside-IV in rat plasma. AB - Astragaloside-IV (3-O-beta-d-xylopyranosyl-6-O-beta-d-glucopyranosyl cycloastragenol) is the major active constituent contained in Radix Astragali. This paper describes a rapid, sensitive and specific assay for quantitative determination of astragaloside-IV in rat plasma. After a liquid/liquid extraction (LLE) with n-butanol and high-performance liquid chromatography (HPLC) gradient separation with acetonitrile-NH4Cl solution (0.5 micromol/L) as the mobile phase, the anions adduct [M + Cl]- at m/z 819.4 of astragaloside-IV, and [M + Cl]- at m/z 815.35 of internal standard (IS) digoxin were analyzed by electrospray ionisation-mass spectrometry (LC-ESI-MS) in selected ion monitoring (SIM) mode. Chromatographic separation was achieved in less than 9 min and calibration curve was linear over a concentration range of 2-200 ng/ml. The described assay method was successfully applied to the preclinical pharmacokinetic study of astragaloside-IV. After intragastric administration of astragaloside-IV to rats, Cmax and Tmax of astragaloside-IV were 134.73 +/- 39.86 ng/ml and 1.5 h, respectively, and the elimination half-life (t1/2) was 5.45 +/- 0.39 h. PMID- 16307860 TI - A new approach to evaluate stability of amodiaquine and its metabolite in blood and plasma. AB - A stability study for amodiaquine (AQ) and desethylamodiaquine (AQm) in whole blood and plasma is reported. AQ, AQm and chloroquine (CQ) were simultaneously analysed and the ratios AQ/CQ and AQm/CQ were used to ensure correct interpretation of the stability results. CQ was stable in whole blood and plasma at all tested temperatures enabling it to be a stability marker in stability studies. Simultaneous analysis of compounds, of which at least one is already known to be stable, permits a within sample ratio to be used as a stability indicator. The new approach significantly reduced bias when compared to the traditional approach. AQ and AQm were stable in plasma at -86 degrees C and -20 degrees C for 35 days, at 4 degrees C for 14 days and at 22 degrees C for 1 day. AQ and AQm were stable in blood at -86 degrees C and 4 degrees C for 35 days, at 20 degrees C and 22 degrees C for 7 days and at 37 degrees C for 1 day. PMID- 16307861 TI - Sensitive determination of G-protein-coupled receptor binding ligands by solid phase extraction-electrospray ionization-mass spectrometry. AB - High affinity Histamine H2-receptor binding ligands were assayed by automated solid phase extraction (SPE) coupled via electrospray ionization with a Quadrupole-Time-of-Flight mass spectrometer (Q-ToF-MS). The mass spectrometric behavior of these analytes was tested in aqueous solutions with several (nine) volatile salts, in different pH, and with various methanol contents. Out of the high amount of available ligands, three fluorescent-labeled molecules (5706, 5707, and 5708) were studied in detail. The limits of detection (LODs) for all three compounds obtained in mass spectrometric detection was 1 fmol (absolute) in continuous flow and FIA (flow injection analysis) measurements. The results obtained with FIA-fluorescence detection gave LODs a factor 10-100 times higher. A systematic investigation of sample solving conditions, loading flow conditions, and elution flow conditions made the automated SPE-MS coupling efficient. Ideally, the ligands were dissolved in MeOH-25 mM phosphate buffer (30:70 v/v; pH 11), the SPE loading flow comprised MeOH-25 mM phosphate buffer (30:70 v/v; pH 11) and the SPE elution flow contained MeOH-100 mM ammonium formate solution (90:10 v/v; pH 3). Using this method on a C18-modified silica cartridge (C18, 5 microm, 100 A, 300 microm i.d. x 5 mm, LC Packings) assures high recovery and achieved LODs for all three compounds of 5 fmol (absolute). As an absolute amount of ligands specifically bound on H2-receptors in biochemical experiments is, as will be published elsewhere, between 10 and 100 fmol, the SPE-MS method for the basic compounds can be directly applied for these Histamine H2-receptors. PMID- 16307862 TI - Characterization of the cytochrome P450 enzymes and enzyme kinetic parameters for metabolism of BVT.2938 using different in vitro systems. AB - An important step in the drug development process is identification of enzymes responsible for metabolism of drug candidates and determination of enzyme kinetic parameters. These data are used to increase understanding of the pharmacokinetics and possible metabolic-based drug interactions of drug candidates. The aim of the present study was to characterize the cytochrome P450 enzymes and enzyme kinetic parameters for metabolism of BVT.2938 [1-(3-{2-[(2-ethoxy-3-pyridinyl)oxy]ethoxy} 2-pyrazinyl)-2(R)-methylpiperazine], a potent and selective 5HT2c-receptor agonist. The enzyme kinetic parameters were determined for formation of three main metabolites of BVT.2938 using human liver microsomes and expressed cytochrome P450 (CYP) isoforms. The major metabolite was formed by hydroxylation of the pyridine ring (CL(int)=27 microl/mgmin), and was catalysed by both CYP2D6*1 and CYP1A1, with K(m) values corresponding to 1.4 and 2.7 microM, respectively. The results from enzyme kinetic studies were confirmed by incubation of BVT.2938 in the presence of the chemical inhibitor of CYP2D6*1, quinidine. Quinidine inhibited the formation of the major metabolite by approximately 90%. Additionally, studies with recombinant expressed CYP isoforms from rat indicated that formation of the major metabolite of BVT.2938 was catalysed by CYP2D2. This result was further confirmed by experiments with liver slices from different rat strains, where the formation of the metabolite correlated with phenotype of CYP2D2 isoform (Sprague-Dawley male, extensive; Dark Agouti male, intermediate; Dark Agouti female, poor metabolizer). The present study showed that the major metabolite of BVT.2938 is formed by hydroxylation of the pyridine ring and catalysed by CYP2D6*1. CYP1A1 is also involved in this reaction and its role in extra-hepatic metabolism of BVT.2938 might be significant. PMID- 16307863 TI - Phosphatase PTEN in chicken muscle is regulated during ontogenesis. AB - The phosphatase and TENsin homolog deleted on chromosome 10 (PTEN) is a lipid and protein phosphatase able to inhibit significant actors of cell signaling (i.e. phosphatidylinositol-3'kinase and mitogen-activated protein kinase pathways). The aim of this study was to characterize PTEN and to investigate its regulation during ontogenesis in chicken muscle. Pectoralis major muscle was sampled on day 18 of the embryonic period (E18), at hatching (d0) and in fed chickens at 2, 7 and 43 days after hatching (d2, d7 and d43). We first cloned the totality of chicken PTEN cDNA; its translation into a putative protein showed more than 95% sequence identity with that characterized in mammals (humans, mice). PTEN was expressed under two major transcripts in the majority of tissues, including muscles where the expression of PTEN mRNA increased with age (P < 0.05). Surprisingly, the protein levels of PTEN (protein characterized with an apparent molecular weight of 55kDa) and its activity were considerably decreased between the E18 and d43 stages (approximately 8-10-fold reduction, P < 0.001). An association between these decreases and higher phosphorylation levels of two potential indirect downstream targets of phosphatase (i.e. AKT and ERK) was observed only in the early growth phases. It was concluded that phosphatase PTEN was expressed in chicken muscle and that its expression was regulated during ontogenesis. PMID- 16307864 TI - The neuropsychological phenotype of velocardiofacial syndrome (VCFS): relationship to psychopathology. AB - Children with velocardiofacial syndrome (VCFS; N=14) and a comparison group of siblings (N=8) underwent comprehensive neuropsychological assessment to examine the relationship between cognitive functioning and psychopathology. Significant group differences were obtained on tests of full scale and verbal intellectual functioning and perceptual-motor skills. With the exception of performance on tests of attention and executive functioning, children with VCFS displayed a profile consistent with nonverbal learning disability (NLD). However, within group comparisons revealed significantly poorer visuospatial intellectual and nonverbal memory functioning in sibling controls as well. No significant group differences were obtained on tests of motor speed, academic, language, attention, memory, or executive functioning, with significant variability in children with VCFS frequently accounting for the lack of robust differences. Parent-report measures revealed profiles consistent with ADHD. No clinically significant symptoms of psychosis, depression or anxiety were noted on either self- or parent report measures. Wisconsin Card Sorting Test performance was found to be highly and negatively correlated with the Thought Problems subscale of the Child Behavior Checklist (CBCL) for VCFS children only, suggesting a possible at-risk indicator for later onset psychopathology. PMID- 16307865 TI - Computational analysis and refinement of sequence structure on chromosome 22q11.2 region: application to the development of quantitative real-time PCR assay for clinical diagnosis. AB - The low-copy repeat (LCR) is a new class of repetitive DNA element and has been implicated in many human disorders, including DiGeorge/velocardiofacial syndrome (DGS/VCFS). It is now recognized that nonallelic homologous recombination (NAHR) through LCRs flanking the chromosome 22q11.2 region leads to genome rearrangements and results in the DGS/VCFS. To refine the structure and content of chromosome 22q11.2 LCRs, we applied computational analysis to dissect region specific LCRs using publicly available sequences. Nine distinct duplicons between 1.6 and 65 kb long and sharing >95% sequence identity were identified. The presence of these sequence motifs supports the NAHR mechanism. Further sequence analysis suggested that the previously defined 3-Mb deletion may actually comprise two deletion intervals of similar size close to each other and thus indistinguishable when using fluorescence in situ hybridization (FISH) analysis. The differentially deleted regions contain several hypothetical proteins and UniGene clusters and may partially explain the clinical heterogeneity observed in DGS/VCFS patients with the 3-Mb common deletion. To implement further sequence information in molecular medicine, we designed a real-time quantitative PCR assay and validated the method in 122 patients with suspected DGS/VCFS. The assay detected 28 patients with chromosome 22q11.2 deletion later confirmed using FISH. Our results indicated that the developed assay is reliable as well as time and cost effective for clinical diagnosis of chromosome 22q11.2 deletion. They also suggest that this methodology can be applied to develop a molecular approach for clinical detection and diagnosis of other genomic disorders. PMID- 16307866 TI - Norman Cousins Lecture. The uses and abuses of psychoneuroimmunology: a global overview. AB - Studies of interactions between the nervous and immune systems that effect immunological and behavioral changes are relevant to our understanding biological issues pertinent to evolution, ethology, ecology, and aging, in addition to our understanding the immune and nervous systems per se. Psychoneuroimmunology also relates to homeland security, science education, and the practice of conventional as well as complementary and alternative medicine. This paper will highlight just some of these global implications of psychoneuroimmunology. PMID- 16307867 TI - Perinatal exposure to endocrine disrupting compounds alters behavior and brain in the female pine vole. AB - Endocrine-disrupting chemicals (EDCs) are synthetic chemicals that arise from sources such as pesticides and have the ability to mimic or inhibit gonadal steroid hormones. The objective of this research was to examine the effects of EDCs on the behaviors associated with monogamy and the expression of related neuropeptide receptors. Pine voles, a novel experimental mammal, were chosen because they display strong monogamous pair bonding. Female pine voles were orally administered estrogenic diethylstilbestrol (DES) and methoxychlor (MXC) or oil control throughout gestation and lactation of pups. Exposed pups were tested as adults. Preference for the mate and maternal behaviors were assessed. While the ability to form partner preferences was intact, DES-exposed females showed increased aggression toward a stranger, while MXC exposed females showed a strong trend toward spending more time alone. Oxytocin (OT) receptor binding in the brain was assessed for possible effects on this behaviorally important neuropeptide signaling system. The cingulate cortex showed a reduction in OT binding in the MXC group. These findings demonstrate that exposure to EDCs during pre- and neonatal development can alter female adult neural phenotype and behavior related to monogamous behavior traits. PMID- 16307868 TI - Spontaneous dyskinesia and familial liability to schizophrenia. AB - Several factors suggest that spontaneous dyskinesia may be a useful supplemental phenotype for further elucidating the specific nature of the genetic contribution to schizophrenia. For example, involuntary movement abnormalities have been observed in both medicated and unmedicated schizophrenia patients, in individuals with schizotypal personality disorder, and sometimes in siblings of schizophrenia patients. However, there are many inconsistencies present in the literature to date. The current study thus sought to investigate the existence of spontaneous dyskinesia in schizophrenia patient probands, their non-psychotic siblings, and healthy controls in order to clarify its potential value as an "endophenotype" in genetic studies of schizophrenia. Videotaped interviews were coded for the presence of spontaneous, involuntary movement abnormalities by a trained and reliable rater using computer assisted technology who was blind to group and family status. The results of this study indicated that siblings of schizophrenia patients did not display significantly more involuntary movements compared to controls, although tremor was observed in a few siblings. In contrast, schizophrenia patients did display significantly more involuntary movements compared to controls as well as their non-psychotic siblings. The lack of significant differences between siblings and controls argues against a strong association between spontaneous dyskinesia and an "unexpressed" genetic liability to schizophrenia. Thus, it appears that involuntary movement abnormalities may be limited in their utility as endophenotypes in genetic studies of schizophrenia, despite being associated with, and perhaps predictive of, schizophrenia itself. PMID- 16307869 TI - Ecophysiology and molecular phylogeny of bacteria isolated from alkaline two phase olive mill wastes. AB - The use of two-phase centrifugal decanters has been widely adopted in the olive oil extraction industry in order to reduce the huge quantities of wastewaters produced during the traditional three-phase extraction process. The resulting sludge-like byproduct, widely known as "alpeorujo", has a pH of 4-6, low water activity (a(w)) and high phytotoxicity. Addition of Ca(OH)(2) to alpeorujo, which is commonly performed at the olive oil mill to handle disposal problems related to acidic pH and odor emissions, creates an alkaline secondary waste (alkaline alpeorujo). Bacteria isolated from alkaline alpeorujo were cultured in order to investigate their physiological and phylogenetic characteristics. The bacterial population at neutral pH was estimated to be 6.0+/-0.4 x 10(7) cells g(-1) dw, while the bacterial population at pH 11 reached 2.1+/-0.3 x 10(5) cells g(-1) dw. Fourteen strains isolated from alkaline pH were halotolerant alkaliphiles, while seven isolates from neutral pH were moderate to extreme halotolerant or/and alkalitolerant bacteria. Based on 16S rRNA gene sequence analysis, four of the halotolerant alkaliphilic isolates showed 98.4-99.2% similarity to known sequences of Bacillus alcalophilus and Nesterenkonia lacusekhoensis, whereas ten isolates demonstrated low percentage similarities (94.4-96.9%) to the genera Idiomarina, Halomonas and Nesterenkonia. As concerns bacteria isolated from neutral pH, four isolates were associated with Corynebacterium, Novosphingobium, Serratia marcescens and Pseudomonas aeruginosa (98.3-99.9% similarities), while three isolates presented 96.5-97.2% sequence similarities to Rhodobacter, Pseudomonas and Ochrobactrum. At least six groups of isolates represent novel phylogenetic linkages among Bacteria. PMID- 16307870 TI - Voltage-dependent anion channel (VDAC) as mitochondrial governator--thinking outside the box. AB - Despite a detailed understanding of their metabolism, mitochondria often behave anomalously. In particular, global suppression of mitochondrial metabolism and metabolite exchange occurs in apoptosis, ischemia and anoxia, cytopathic hypoxia of sepsis and multiple organ failure, alcoholic liver disease, aerobic glycolysis in cancer cells (Warburg effect) and unstimulated pancreatic beta cells. Here, we propose that closure of voltage-dependent anion channels (VDAC) in the mitochondrial outer membrane accounts for global mitochondrial suppression. In anoxia, cytopathic hypoxia and ethanol treatment, reactive oxygen and nitrogen species, cytokines, kinase cascades and increased NADH act to inhibit VDAC conductance and promote selective oxidation of membrane-permeable respiratory substrates like short chain fatty acids and acetaldehyde. In cancer cells, highly expressed hexokinase binds to and inhibits VDAC to suppress mitochondrial function while stimulating glycolysis, but an escape mechanism intervenes when glucose-6-phosphate accumulates and dissociates hexokinase from VDAC. Similarly, glucokinase binds mitochondria of insulin-secreting beta cells, possibly blocking VDAC and suppressing mitochondrial function. We propose that glucose metabolism leads to glucose-6-phosphate-dependent unbinding of glucokinase, relief of VDAC inhibition, release of ATP from mitochondria and ATP-dependent insulin release. In support of the overall proposal, ethanol treatment of isolated rat hepatocytes inhibited mitochondrial respiration and accessibility to adenylate kinase in the intermembrane space, effects that were overcome by digitonin permeabilization of the outer membrane. Overall, these considerations suggest that VDAC is a dynamic regulator, or governator, of global mitochondrial function both in health and disease. PMID- 16307871 TI - Event-related potentials to violations of inflectional verb morphology in English. AB - Event-related brain potentials were recorded to morphologically correct and incorrect regular and irregular past tense verb forms presented in sentences and in lists. In the sentence context, all incorrect verb forms elicited a broadly distributed late posterior positivity, as well as a left anterior negativity (LAN) that was particularly pronounced for the incorrect irregulars. Using a single-word paradigm, we did not find a LAN for any of the incorrect verb forms but found an N400-like effect for all irregular verbs. In the sentence context, only the incorrect irregulars elicited a long-lasting, broadly distributed late positivity, reminiscent of the P600. For regular verbs, responses to incorrect forms produced smaller, more time restricted effects. These data show that morphological and syntactic violations produce similar patterns of brain activity, suggesting that these two systems engage cognitive processes with similar underlying neural substrates. PMID- 16307872 TI - Increased DNA damage in patients with chronic obstructive pulmonary disease who had once smoked or been exposed to biomass. AB - OBJECTIVE: Chronic obstructive pulmonary disease (COPD) is a slowly progressive condition characterised by poorly reversible airflow limitation associated with an abnormal inflammatory response of the lung. The main causal factors of COPD are chronic oxidative stress as a result of long-term smoking, use of biomass fuels, and air pollution. In this study, basal levels of DNA strand breaks were investigated together with some additional oxidative markers implicating oxidative damage on the other biomolecules such as proteins and lipids in patients with COPD who were exposed to smoking and biomass. MATERIAL AND METHODS: We detected DNA strand breaks in peripheral blood mononuclear leukocytes by using a Single Cell Gel Electrophoresis (also called Comet Assay), plasma protein carbonyl (PC) content by using Reznick and Parker's spectrophotometric method, and lipid peroxidation by measurement of malondialdehyde (MDA) as indexes of oxidative stress in 47 patients with smoking-related COPD and 25 patients with biomass-related COPD and 36 age-and-sex matched control participants. RESULTS: The mean values of DNA strand breaks, MDA and protein carbonyl levels were significantly higher in smoking- and biomass-related COPD groups than in the control group (ANOVA P<0.001, <0.05 and <0.05, respectively). DNA damage levels were also higher in smoking-related COPD group than in biomass-related COPD group (P<0.05). There was a positive relationship between DNA damage and MDA levels in smoking-related COPD group (P<0.05). CONCLUSION: Oxidative stress markers and DNA damage were strongly increased in both patient groups with smoking- and biomass related COPD. However, DNA is more affected in smoking-related COPD patients than in biomass-related COPD. These data indicate that cigarette smoking is a more significant DNA damaging risk factor than biomass smoke. PMID- 16307873 TI - Effects of composted sewage sludge on microbial biomass, activity and pine seedlings in nursery forest. AB - The investigation was carried out in a 2 year experiment to evaluate the benefits and hazards of the use of composted sewage sludge as a restoration agent for the soil of the nursery forest intended for growing Pinus sylvestris seedlings. The grey forest soil (Haplic Greyzem) was amended with compost at the 25, 50, 75, 100, 150 and 175 t ha(-1) application rates on a dry matter basis. The organic matter content increased with the increase in sludge amendment as well as the metal content. However, the concentrations of individual metals were below the current limits established for Russia and European countries. Sludge amendments enhanced the germination and decreased the mortality of the seedlings. The effects were more obvious for the soil with the highest sludge treatment. The beneficial effects on the biomass of seedlings and the height of the shoots as well as on the length of the roots of the pine seedlings were greater in plots with the highest rates of composted sludge. The application of composted sludge to soil was followed by an increase in microbial biomass and to a lesser extent in basal respiration. In the absence of any detrimental effect on microorganisms, this study lends support to using composted sewage sludge as the organo-mineral fertilizer for the soil of nursery forest. PMID- 16307874 TI - Application of large volume injection GC-MS to analysis of organic compounds in the extracts and leachates of municipal solid waste incineration fly ash. AB - Organic solvent and water extracts of fly ash from a Milan (Italy) municipal solid waste incinerator (MSWI) were analyzed by large volume injection-gas chromatography-mass spectrometry (LVI-GC-MS) with programmable temperature vaporizer (PTV). Using injection volumes of 10-100 microl, typically over a hundred compounds were detected in organic solvent extracts and ca. 35% of them could be tentatively identified from their electron impact ionization mass spectra. A protocol for the determination of the maximum amount of a potential environmental pollutant available for leaching (availability test) was developed for four selected target compounds: pentachlorobenzene (PeCB), hexachlorobenzene (HxCB), o-terphenyl (o-TPH) and m-terphenyl (m-TPH). Key parameters, extraction time and liquid-to-solid ratio (L/S), were studied in more detail. Recoveries of PeCB, HxCB and o-TPH spiked into the fly ash samples at two concentration levels ranged from 38% to 53% for freshly spiked and from 14% to 40% for 40-day aged fly ash. Recoveries of m-TPH were 8% to 11% from freshly spiked and less than 3% from aged spiked fly ash. The native amounts in Milan MSWI fly ash, determined in an interlaboratory exercise using the developed protocol, were 31 ng/g PeCB, 34 ng/g HxCB, 72 ng/g o-TPH and 4.4 ng/g m-TPH. A separate methodology was developed for the determination of compounds extracted from fly ash by water (leaching test). Following 8-h sonication at L/S 20, the leached amounts of PeCB, HxCB and o-TPH were 1.1, 3.1 and 6.0 ng/g fly ash, respectively. PMID- 16307875 TI - Extraction of heavy metal ions from waste colored glass through phase separation. AB - A new method utilizing phase separation phenomena for the extraction of heavy metal ions used as colorants in colored glass is proposed. Colored soda-lime silica glass containing Co or Cr as a colorant was remelted with B2O3 to yield soda-lime-borosilicate glass. The soda-lime-borosilicate glass thus obtained was leached in 1M nitric acid at 90 degrees C to dissolve the borate phase. All cations (Na, Ca, Cr and Co) concentrated in the borate phase are successfully leached out with the dissolution of the borate phase, when the amount of the B2O3 added to the glass and heat treatment conditions are properly chosen. Porous silicate glass powders with high SiO2 purity are obtained as the result of the leaching. Porous glass can also be formed as bulk material by controlling the composition of additives during the remelting. PMID- 16307877 TI - Improvement of the quality of whole wheat bread by supplementation of xylanase from Aspergillus foetidus. AB - The possible use of xylanase from Aspergillus foetidus MTCC 4898 as a bread improver was tested in whole wheat bread. The partially purified xylanase was used as an additive at 12 U/g during mixing of wheat flour. The effects of xylanase addition on the fermentation stage and the final bread quality were analyzed. Remarkable decrease (11%) in water absorption and increase in dough rising (28.5%) were noticed. Final moisture content of the bread was higher (40.5%) than control (32.3%). Improvements in volume (53%) and specific volume (56%) were also significant. Sensory evaluation indicated better flavour, taste, softness and overall acceptability. Texture profile analysis confirmed the rheological changes. Firmness was decreased by more than four folds. Improvements in cohesiveness and decline in springiness and gumminess were observed. PMID- 16307878 TI - Synthesis and structure-activity relationships of 8-azabicyclo[3.2.1]octane benzylamine NK1 antagonists. AB - A series of 8-azabicyclo[3.2.1]octane amine hNK1 antagonists has been investigated and structure-activity relationships of the benzylamine and 6-exo substituents described. Acidic substituents at C6 give a series of high affinity compounds for hNK1 with selectivity over the hERG channel. PMID- 16307879 TI - Structure-guided synthesis of tamoxifen analogs with improved selectivity for the orphan ERRgamma. AB - The design and synthesis of 4-hydroxytamoxifen (4-OHT) derivatives are described. The binding affinities of these compounds toward the orphan estrogen-related receptor gamma and the classical estrogen receptor alpha demonstrate that analogs bearing hydroxyalkyl groups display improved binding selectivity profiles compared with that of 4-OHT. An X-ray crystal structure of one of the designed compounds bound to ERRgamma LBD confirms the molecular basis of the selectivity. PMID- 16307880 TI - Tacrolimus in refractory patients with myasthenia gravis: coadministration and tapering of oral prednisolone. AB - We prospectively investigated therapeutic and adverse effects of tacrolimus in seven patients with myasthenia gravis (MG) who were resistant to conventional therapies or could not be treated with thymectomy because of complications. Within two months of initiation of tacrolimus all the patients subjectively showed improvement of clinical symptoms, while both the quantitative MG score for disease severity and MG-activities of daily living profile were significantly decreased (p<0.05) 3 and 6 months after commencement compared with before. Nine months after initiation and later, MG temporarily exacerbated in two patients with rapid tapering of oral prednisolone and one non-thymectomized one. This drug is useful in the treatment of refractory patients with MG irrespective of thymectomy, particularly in the early phase after commencement. When tacrolimus is additionally used for treatment of MG, preceding drugs, particularly corticosteroids such as oral prednisolone, should be carefully tapered if necessary in order to prevent clinical exacerbation. PMID- 16307881 TI - Tricyclic pharmacophore-based molecules as novel integrin alpha(v)beta3 antagonists. Part III: synthesis of potent antagonists with alpha(v)beta3/alpha(IIb)beta3 dual activity and improved water solubility. AB - In order to optimize our novel integrin alpha(v)beta3/alpha(IIb)beta3 dual antagonists, spatial screening at the N-terminus was performed. The alpha(v)beta3 antagonistic activity varied depending on the space that was occupied by the N terminus, but high potency against alpha(IIb)beta3 was well maintained. The (3S) aminopiperidine analogue had the strongest activity against alpha(v)beta3, and the S isomer at piperidine was more potent than the R isomer. Compounds selected on the basis of SAR analysis of a novel lead compound showed acceptable early absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles and sufficient water solubility for use as infusion drugs. Docking studies with the alpha(v)beta3 receptor were performed to confirm the SAR findings. PMID- 16307882 TI - Discovering novel chemical inhibitors of human cyclophilin A: virtual screening, synthesis, and bioassay. AB - Cyclophilin A (CypA) is a member of cyclophilins, a family of the highly homologous peptidyl prolyl cis-trans isomerases (PPIases), which can bind to cyclosporin A (CsA). CypA plays critical roles in various biological processes, including protein folding, assembly, transportation, regulation of neuron growth, and HIV replication. The discovery of CypA inhibitor is now of a great special interest in the treatment of immunological disorders. In this study, a series of novel small molecular CypA inhibitors have been discovered by using structure based virtual screening in conjunction with chemical synthesis and bioassay. The SPECS_1 database containing 85,000 small molecular compounds was searched by virtual screening against the crystal structure of human CypA. After SPR-based binding affinity assay, 15 compounds were found to show binding affinities to CypA at submicro-molar or micro-molar level (compounds 1-15). Seven compounds were selected as the starting point for the further structure modification in considering binding activity, synthesis difficulty, and structure similarity. We thus synthesized 40 new small molecular compounds (1-6, 15, 16a-q, 17a-d, and 18a l), and four of which (compounds 16b, 16h, 16k, and 18g) showed high CypA PPIase inhibition activities with IC50s of 2.5-6.2 microM. Pharmacological assay indicated that these four compounds demonstrated somewhat inhibition activities against the proliferation of spleen cells. PMID- 16307883 TI - Structure-activity relationships of a series of tariquidar analogs as multidrug resistance modulators. AB - Tariquidar (XR9576) analogs, modulators of cancer multidrug resistance (MDR), were subjected to QSAR and 3D-QSAR analyses. The structural features contributing to anti-MDR activity were identified by the Free-Wilson analysis and pharmacophore search using Hoechst 33342 as a template. 3D-QSAR CoMFA and CoMSIA models were derived and tested. The best models yielded an external predictivity of 0.66-0.75 squared correlation coefficient and outlined HB-acceptor, steric, and hydrophobic fields as the most important 3D properties. On the basis of the QSAR and 3D-QSAR analyses it was suggested that the strong inhibitory potency of the compounds studied is related to the presence of a bulky aromatic ring system with a 3rd positioned heteroatom toward the anthranilamide nucleus in the opposite end of the tetrahydroquinoline group. The results can help in directing the rational design of new generations of potent P-glycoprotein MDR modulators. PMID- 16307884 TI - Macrocyclic inhibitors of the malarial aspartic proteases plasmepsin I, II, and IV. AB - The first macrocyclic inhibitor of the Plasmodium falciparum aspartic proteases plasmepsin I, II, and IV with considerable selectivity over the human aspartic protease cathepsin D has been identified. A series of macrocyclic compounds were designed and synthesized. Cyclizations were accomplished using ring-closing metathesis with the second generation Grubbs catalyst. These compounds contain either a 13-membered or a 16-membered macrocycle and incorporate a 1,2 dihydroxyethylene as transition state mimicking unit. The binding mode of this new class of compounds was predicted with automated docking and molecular dynamics simulations, with an estimation of the binding affinities through the linear interaction energy (LIE) method. PMID- 16307885 TI - Novel vitamin D3 antipsoriatic antedrugs: 16-En-22-oxa-1alpha,25-(OH)2D3 analogs. AB - A series of 16-en-22-oxa-derivatives of vitamin D3 based on the structure of maxacalcitol (2) were prepared. Maxacalcitol is currently used topically for the treatment of psoriasis and is recognized as the most successful antedrug of natural vitamin D(3) because it retains the original antiproliferative activity of calcitriol without increased calcemic activity. We introduced 16-olefinic functionality to accelerate the oxidative metabolism of the drug in liver, presumed to be essential for the reduction of calcemic activity, and modified the side-chain moiety by placing the 22-oxygen on the more labile allylic carbon center. Novel 22-oxa analogs (7a-i), carrying either the 24-alkynyl bond or 24 hydroxy functionality in addition to the 16-double bond were synthesized and their pharmacokinetics were evaluated. PMID- 16307886 TI - 60Co aqueous speciation and pH effect on the adsorption behavior on inorganic materials. AB - The adsorption of radioactive cobalt from aqueous solution on MgO, MnO(2), TiO(2), SnO, activated carbon and calcined hydrotalcite was studied under static conditions as a function of pH (1, 3, 5, 7, and 10) of the (60)Co solution. It was found that under the experimental conditions, (60)Co was significantly adsorbed on MnO(2), TiO(2) and activated carbon and in smaller quantities on MgO, SnO and calcined hydrotalcite. The adsorption of (60)Co species on MgO, MnO(2), TiO(2) and SnO can be attributed to the direct exchange of ions with a specific group on the adsorbing surface, in accordance with the concept of zero point charge (zpc). The radionuclide content was determined by gamma-spectrometry. The (60)Co chemical species present in aqueous solutions with different pH values were surveyed by using high-voltage electrophoresis. It was also found that (60)Co was present only as a cationic species at pH 1, 3, 5 and 7 and a neutral species at alkaline pH. PMID- 16307887 TI - Radiosynthesis of (E)-N-(2-[11C]methoxybenzyl)-3-phenyl-acrylamidine, a novel subnanomolar NR2B subtype-selective NMDA receptor antagonist. AB - Recently, a novel series of amidines has been described, exhibiting high NR2B subtype selective N-methyl-D-aspartate (NMDA) antagonist activity with nanomolar or subnanomolar affinity. Within the styrylamidine subclass, (E)-N-(2 methoxybenzyl)-3-phenyl-acrylamidine (1), displayed the highest affinity (Ki=0.7 nM versus [(3)H]ifenprodil) and was considered an appropriate candidate for isotopic labelling with carbon-11 (T(1/2): 20.38 min) at its methoxy group for imaging of NMDA receptors with PET. Derivative 1 has been labelled from the corresponding nor-analogue using [(11)C]methyl triflate and the following experimental conditions : (1) trapping at -10 degrees C of [(11)C]methyl triflate in 300 microL of acetone containing 0.6-0.8 mg of precursor 5 (2.4-3.2 micromol) and 5 microL of a 3M solution of NaOH in water (about 5 eq.); (2) concentration to dryness of the reaction mixture (at 110 degrees C, using a helium stream for 1 2 min); (3) taking up the residue with 0.5 mL of the HPLC mobile phase and (4) purification using semi-preparative HPLC (SymmetryPrep) C-18, Waters, 300 x 7.8 mm). Typically, starting from a 1.5 Ci (55.5 GBq) [(11)C]CO(2) production batch, 120-240 m Ci (4.44-8.88 GBq) of [(11)C]-1 (20-40% decay-corrected radiochemical yield, n=5) was obtained within a total synthesis time of 25-30 min. Specific radioactivities ranged from 0.8 to 1.2 Ci/micromol (29.6-44.4 GBq/micromol) at the end of radiosynthesis. No attempts were made to further optimise these reactions, as sufficient material was obtained to allow for preliminary pharmacological characterisation. PMID- 16307888 TI - Effect of detector collimator and sample thickness on 0.662 MeV multiply Compton scattered gamma rays. AB - The simultaneous effect of detector collimator and sample thickness on 0.662 MeV multiply Compton-scattered gamma photons was studied experimentally. An intense collimated beam, obtained from 6-Ci (137)Cs source, is allowed to impinge on cylindrical aluminium samples of varying diameter and the scattered photons are detected by a 51 mm x 51 mm NaI(Tl) scintillation detector placed at 90 degrees to the incident beam. The full energy peak corresponding to singly scattered events is reconstructed analytically. The thickness at which the multiply scattered events saturate is determined for different detector collimators. The parameters like signal-to-noise ratio and multiply scatter fraction (MSF) have also been deduced and support the work carried out by Shengli et al. [2000. EGS4 simulation of Compton scattering for nondestructive testing. KEK proceedings 200 20, Tsukuba, Japan, pp. 216-223] and Barnea et al. [1995. A study of multiple scattering background in Compton scatter imaging. NDT & E International 28, 155 162] based upon Monte Carlo calculations. PMID- 16307889 TI - Dynamic regulation of microglial functions by the non-steroidal anti-inflammatory drug NCX 2216: implications for chronic treatments of neurodegenerative diseases. AB - The nitric oxide-releasing derivative of flurbiprofen, NCX 2216, has a safer gastrointestinal profile than the parent drug flurbiprofen and a strong anti amyloidogenic activity. Here, we show that in primary microglial cultures, in addition to the expected inhibition of prostaglandin synthesis, NCX 2216 specifically activated the peroxisome proliferator-activated receptor-gamma (PPAR gamma), a ligand-dependent transcription factor controlling several important microglial functions. Prolonged treatment (16 h) of microglial cultures with NCX 2216 induced PPAR-gamma nitration and prevented further activation of the receptor by specific agonists. At functional levels, NCX 2216 treatment of LPS activated microglial cultures resulted in the transient reduction of TNF-alpha and NO production and in the protracted inhibition of IL-1beta and PGE2 synthesis. The dynamic regulation of the functional state of activated microglia by NCX 2216 helps explaining recent findings in Alzheimer's disease animal models and may offer new therapeutic opportunities for treating neurodegenerative diseases. PMID- 16307890 TI - Quantitative spectrophotometry using integrating cavities. AB - Absorption spectrophotometry, a standard tool for quantitative analysis, suffers from two major drawbacks: lack of sensitivity and vulnerability to scattering. It has been pointed out earlier that the solution to these problems lies in using a reflecting cavity as a sample holder. Due to multiple reflections at the cavity wall, the effective pathlength becomes considerably larger than the diameter of the cavity, and scattering losses are eliminated because scattered light is prevented from escaping the detector. Though much effort has been spent in analysing and improving the performance of such a device, often called an integrating cavity absorption meter (ICAM), a simple strategy for deducing the absorbance of the sample is still lacking. It is shown here that the absorbance A' measured by using an ICAM exhibits a sublinear increase with the solute concentration C. The physical reason for this departure from linearity is explained, and a straightforward procedure for converting A' to the true absorbance A (proportional to C) is established. The reliability of the procedure is demonstrated by comparing the ICAM absorption spectrum of dilute dye solutions with the spectra of more concentrated solutions recorded in a conventional spectrophotometer. The ability of the device to cope with scattering was tested by filling the ICAM with a suspension of chloroplasts, and the spectrum was found, as expected, to be free from scattering artefacts. PMID- 16307891 TI - Putting the brakes on lactotrope hyperplasia. AB - Although much is known about the mechanisms by which dopamine, the major regulator of lactotrope function, controls hormone production and secretion, its growth inhibiting actions have remained less well characterized. Recent research has uncovered an important role for endogenously produced transforming growth factor beta (TGFbeta) and TGFbeta receptors in dopamine's ability to put the brakes on lactotrope proliferation. These novel findings promise increased knowledge of the genesis and potential therapeutic control of human prolactinomas. PMID- 16307892 TI - The kappa-like pre-B receptor: surplus biology or a missing link? AB - After the demonstration that surrogate JCkappa polypeptides could covalently bind mu heavy chain and upon the characterization of the Vkappa-like component of the kappa-like pre-B cell receptor, it became evident that germline transcription is not sterile. The present review discusses the concept of the alternative usage of kappa-like pre-B cell receptors and classical pre-B cell receptors utilizing the lambda-like surrogate light chain composed of lambda5 and VpreB. We propose that both kappa-like and lambda-like pre-B cell receptors work in concert in a fail safe mechanism to promote light chain rearrangement, heavy chain allelic exclusion and B-lymphocyte maturation. PMID- 16307893 TI - Integrin alpha3beta1 suppresses long-term potentiation at inhibitory synapses on the cerebellar Purkinje neuron. AB - At the GABAergic synapses between inhibitory interneurons and a Purkinje neuron (PN) in the cerebellum, the postsynaptic depolarization induces the long-term potentiation (called rebound potentiation; RP) of GABAA receptor responsiveness. Here, we show that integrins, a type of cell-adhesion molecules, are involved in the regulation of RP. Integrin activation by Mn2+ impaired the RP induction of GABA responsiveness and mIPSCs in PNs, which was abolished by the function blocking antibody against either integrin alpha3 or beta1 subunit, but not by that against alpha5 or alphaV subunit. Furthermore, overexpression of integrin alpha3 subunit in a PN by itself impaired the RP induction. We also show that Src family of protein tyrosine kinases mediated the suppressive effect of integrin activity on the RP induction. Thus, the integrin/Src pathway negatively regulates the induction of long-term plasticity at inhibitory synapses on a cerebellar PN. PMID- 16307894 TI - Efficient solubilization, activation, and purification of recombinant Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli. AB - A cytotoxic protein Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli was solubilized in 10 mM HCl. Protein concentration of saturated solution of the recombinant Cry45Aa in 10 mM HCl was about 25 times higher than that in the buffer of previous method (in 50 mM sodium carbonate buffer, pH 10.5, containing 1 mM EDTA, and 10 mM dithiothreitol). The Cry45Aa solubilized in the acidic solution was activated by pepsin as an alternative to proteinase K in the previous method. Cytotoxic activity against CACO-2 cells of the pepsin-treated Cry45Aa was almost identical to the proteinase K-treated protein. The pepsin-treated Cry45Aa was purified by cation-exchange chromatography. The concentration of the purified protein was 539 microg/ml, which was 27-fold higher than that of the activated Cry45Aa by the previously method. The cytotoxic activity of the purified protein was stable in broad pH region (pH 2.0-11.0) for 3 days, and 97% cytotoxic activity remained after incubation at 30 degrees C for 360 min. PMID- 16307895 TI - Doing the right thing: a common neural circuit for appropriate violent or compassionate behavior. AB - Humans have a considerable facility to adapt their behavior in a manner that is appropriate to social or societal context. A failure of this ability can lead to social exclusion and is a feature of disorders such as psychopathy and disruptive behavior disorder. We investigated the neural basis of this ability using a customized video game played by 12 healthy participants in an fMRI scanner. Two conditions involved extreme examples of context-appropriate action: shooting an aggressive humanoid assailant or healing a passive wounded person. Two control conditions involved carefully matched stimuli paired with inappropriate actions: shooting the person or healing the assailant. Surprisingly, the same circuit, including the amygdala and ventromedial prefrontal cortex, was activated when participants acted in a context-appropriate manner, whether being compassionate towards an injured conspecific or aggressive towards a violent assailant. The findings indicate a common system that guides behavioral expression appropriate to social or societal context irrespective of its aggressive or compassionate nature. PMID- 16307896 TI - The level of spinal cord involvement influences the pattern of movement associated cortical recruitment in patients with isolated myelitis. AB - Using fMRI, an increased recruitment of the ipsilateral primary sensorimotor cortex (SMC), supplementary motor area, and middle frontal gyrus has been detected in patients with cervical cord myelitis of possible demyelinating origin. The aim of this study was to evaluate, using fMRI, whether the level of cord involvement influences cortical reorganization by comparing patients with isolated myelitis of the cervical and the dorsal portions of the cord, and to investigate whether the extent of cortical reorganization is associated with the extent of cervical cord pathology measured using magnetization transfer (MT) MRI. We studied 24 right-handed patients (14 with a previous involvement of the cervical cord and 10 with an involvement of the dorsal cord) in a chronic and clinically stable phase following an isolated myelitis of possible demyelinating origin and 15 sex- and age-matched healthy controls. During a single session, we obtained fMRI during repetitive flexion-extension of the last four fingers of the right and left hands and cervical cord MT MRI. Average cord MTR was lower in patients with cervical (P < 0.0001) and dorsal (P = 0.0001) myelitis than in controls. Compared to controls and for both tasks, patients with myelitis had an increased recruitment of the ipsilateral primary SMC, which was independent of the level of cord involvement. On the contrary, patients with cervical myelitis had a more widespread recruitment of frontal and parietal regions, whereas those with dorsal myelitis had a more widespread recruitment of temporal and cerebellar regions. Strong correlations (r values ranging from -0.72 to -0.88) were found between relative activations of cortical areas and the severity of cervical cord damage. Patients with isolated myelitis have different patterns of movement associated cortical activations according to the level of cord involvement. This "level-dependent" functional reorganization of the cortex is likely to have an adaptive role in limiting the clinical outcome of cord damage and should be considered when designing rehabilitation strategies for these patients. PMID- 16307897 TI - Triple-quantum correlation NMR experiments in solids using J-couplings. AB - We show that triple-quantum-single-quantum (TQ-SQ) correlation spectra of crystalline and disordered solids can be obtained under MAS using pulse sequences based on through-bond J-couplings. The feasibility of the experiments in coupled spin-1/2 systems is demonstrated for fully 13C-labelled L-alanine and Pb3P4O13 crystalline compounds, considered as model three-spin and four-spin systems, respectively. In the case of phosphate glasses, we show that the obtained TQ-SQ correlation spectra provide an improved description of the glass forming network connectivities and of the chain length distribution in the disordered network. PMID- 16307898 TI - Expression study of mutant cystathionine beta-synthase found in Japanese patients with homocystinuria. AB - Cystathionine beta-synthase (CBS) deficiency is the most common cause of homocystinuria. More than 130 pathogenic mutations, mostly in the Caucasian populations, have been described. Recently, our group reported a mutation analysis of Japanese homocystinuric patients. In the present paper, we report an expression study of several mutant CBS enzymes in Escherichia coli, i.e., R121H, G148R, G151R, S217F, H232D, R266G, 1591delTTCG, and K441X. All of the mutants except K441X exhibited severely decreased activity, and the capability to form tetramers of most mutants was severely impaired. The K441X mutant, on the other hand, exhibited relatively high activity (63% of the wild type activity). This was probably due to two factors. First, the high abundance of the full-length CBS protein, a likely K441Q mutant, which was produced through suppression of the amber termination codon by glutamine tRNA in E. coli. And second, the presence of a C-terminally truncated protein, which was previously shown to be constitutively activated. Patient-derived lymphocytes, however, showed no detectable CBS subunits. As previously hypothesized, the increased aggregation of mutant CBS subunits might be a common pathogenic mechanism in CBS deficiency. PMID- 16307899 TI - Near infrared emission spectroscopy induced by ultrasonic irradiation. AB - Near infrared emission caused by ultrasonic excitation is demonstrated for the first time in this work. The instrument is constituted of an acousto-optical tunable filter-based spectrometer, an ultrasonic processor connected to a titanium alloy ultrasonic probe and a cylindrical borosilicate flask containing the sample to be excited. The radiation emitted by the sample is collected by a concave mirror and sent to the spectrometer. The effects of the position of the probe extremity in relation to a lateral entrance of the borosilicate flask and of the ultrasonic power on the emission signal were studied. The best results were obtained by positioning the probe extremity up to 2mm from the reflexive body (lateral entrance) using 30% of the full ultrasonic incident power and acquiring spectra after 5 min of sonication. The NIR emission spectra resulting from the ultrasonic excitation were in agreement with that obtained by thermal excitation. The proposed technique was utilized to study different poly(dimethylsiloxane) samples having different viscosities. PMID- 16307900 TI - Intensity non-uniformity correction in MRI: existing methods and their validation. AB - Magnetic resonance imaging is a popular and powerful non-invasive imaging technique. Automated analysis has become mandatory to efficiently cope with the large amount of data generated using this modality. However, several artifacts, such as intensity non-uniformity, can degrade the quality of acquired data. Intensity non-uniformity consists in anatomically irrelevant intensity variation throughout data. It can be induced by the choice of the radio-frequency coil, the acquisition pulse sequence and by the nature and geometry of the sample itself. Numerous methods have been proposed to correct this artifact. In this paper, we propose an overview of existing methods. We first sort them according to their location in the acquisition/processing pipeline. Sorting is then refined based on the assumptions those methods rely on. Next, we present the validation protocols used to evaluate these different correction schemes both from a qualitative and a quantitative point of view. Finally, availability and usability of the presented methods is discussed. PMID- 16307901 TI - A novel mutagen, 2-(5-hydroxy-4,6-dinitroindolyl) ethanol, formed in the reaction between 5-hydroxytryptamine and nitrite under acid conditions, especially in the presence of L-cysteine. AB - We examined the mutagenic activity of each of 29 amino acids mixed under acidic conditions with 5-hydroxytryptamine (5-HT) and nitrite using Salmonella typhimurium strain TA 100 with or without a metabolic activation system (S9 mix). The reaction mixture containing L-cysteine was strongly mutagenic without S9 mix. We subjected an ethyl acetate extract of the reaction mixture to HPLC, isolated a mutagenic component, and investigated its chemical structure by LC-mass spectrometry (MS), high-resolution fast atom bombardment (HRFAB)-MS, and 1H and 13C NMR. We identified the mutagen as 2-(5-hydroxy-4,6-dinitro-3-indolyl) ethanol (2HDIE). We injected 8 mg/kg 2HDIE i.p. into male ICR mice and found that the compound increased the frequency of micronuclei in peripheral reticulocytes. Our results suggest that 2HDIE might be formed in vivo by consumption of 5-HT, nitrite and L-cysteine in foods, and might act as a mutagen. PMID- 16307902 TI - Eye-wiping test: a sensitive animal model for acute trigeminal pain studies. AB - The possibility of introducing eye-wiping test as a model of acute pain was examined in rat, and it was compared with the well-known hot plate test. One drop of NaCl 5 M was placed into the animal eye, and the number of eye wipes with the ipsilateral forelimb was counted during 30 s. The withdrawal latency in hot plate test was also examined. Afterward, animals were treated with morphine (1, 2, 4, 6, 8 or 10 mg/kg), imipramine (25 mg/kg), sodium salicylate (250 mg/kg) or saline (i.p). After 30 min, the animals were tested again with eye-wiping and hot plate tests. Our results showed that morphine injection dose dependently decreased the number of eye wipes and increased the response latency to hot plate tests. There was a good correlation between the analgesic effects of morphine on responses to both tests, however, morphine produced more pain relief in eye-wiping test. Imipramine significantly decreased the number of eye wipes and increased the response latency to hot plate test, while sodium salicylate and saline injection did not. It may be concluded that the eye-wiping test can be used as a reliable method in trigeminal pain studies, which is sensitive to opioid and tricyclic antidepressant in rat. PMID- 16307903 TI - Synthesis, characterization and biological activity of ternary copper(II) complexes containing polypyridyl ligands. AB - Ternary copper(II) complexes involving polypyridyl ligands in the coordination sphere of composition [Cu(tpy)(phen)](ClO4)2 (1), [Cu(tpy)(bipy)](ClO4)2 (2), [Cu(tptz)(phen)](ClO4)2 (3) and [Cu(tptz)(bipy)](BF4)2 (4) where tpy = 2,2':6',2''-terpyridine, tptz = 2,4,6-tri(2-pyridyl)-1,3,5-triazine, phen = 1,10 phenanthroline and bipy = 2,2'-bipyridine have been synthesized and characterized by elemental analysis, magnetic susceptibility, X-band e.p.r. spectroscopy and electronic spectroscopy. Single crystal X-ray of (1) has revealed the presence of a distorted square pyramidal geometry in the complex. Magnetic susceptibility measurements at room temperature were in the range of 1.77-1.81 BM. SOD and antimicrobial activities of these complexes were also measured. Crystal data of (1): P-1, a = 9.3010(7) A, b = 9.7900(6) A, c = 16.4620(6) A, Vc = 1342.73(14) A3, Z = 4. The bond distance of CuN in square base is 2+/-0.04 A. PMID- 16307904 TI - Copper(II)-selective fluorimetric bulk optode membrane based on a 1-hydroxy-9,10 anthraquinone derivative having two propenyl arms as a neutral fluorogenic ionophore. AB - A new optical chemical sensor has been developed for the selective determination of copper(II) ions in aqueous solutions. The reversible sensing system was prepared by incorporating 1-hydrpxy-2-(prop-2'-enyl)-4-(prop-2'-enyloxy)-9,10 anthraquinone (AQ) as a neutral Cu2+-selective fluoroionophore in the plasticized PVC membrane with potassium tetrakis(p-chlorophenyl borate) as an anionic additive. The response of the sensor is based on the fluorescence quenching of AQ by Cu2+ ions. At a pH 5.5, the proposed sensor displays a calibration response for Cu2+ over a wide concentration rang of 1.0 x 10(-2) to 1.0 x 10(-6) M, with a relatively fast response of less than 40 s. In addition to high stability and reproducibility, the sensor shows a unique selectivity towards Cu2+ ion with respect to common co-existing cations. The proposed fluorescence optode was applied successfully to the determination of copper(II) in black tea samples. PMID- 16307905 TI - Cardiac PPARalpha expression in patients with dilated cardiomyopathy. AB - BACKGROUND: The peroxisome proliferator-activated receptor alpha (PPARalpha) is a central regulator of myocardial fatty acid (FA) metabolism implicated in the pathogenesis of heart failure. AIMS: To characterize PPARalpha regulation in human dilated cardiomyopathy (DCM), we studied the expression of cardiac PPARalpha, cardiac carnitine palmitoyl-transferase I (CPT-1), a major PPARalpha target gene, and of the cardiac glucose transporter GLUT-4 in patients with DCM. METHODS: Left ventricular biopsies were taken from patients with DCM (n=16) and control subjects (n=15), and mRNA expression was quantitated using real-time PCR (SYBR((R))Green) and protein expression was measured by Western immunoblotting. RESULTS: Left ventricular PPARalpha mRNA levels were significantly increased in the DCM group compared to the control group (136+/-25.4% vs. control, p<0.01). Consistently, DCM patients had a significantly higher cardiac CPT-1 mRNA expression (147+/-51% vs. control, p<0.05) compared to the control group. Cardiac GLUT-4 expression was similar in both groups. CONCLUSION: Elevated cardiac PPARalpha levels followed by an induction of cardiac CPT-1 expression may result in increased fatty acid metabolism for cardiac energy production in DCM, suggesting a specific cardiac metabolic program in human DCM compared to other types of cardiomyopathy. PMID- 16307906 TI - Onchocerciasis-related epilepsy? Prospects at a time of uncertainty. AB - Epilepsy and onchocerciasis (river blindness) constitute serious public health problems in several tropical countries. There are four main mechanisms that might explain a relationship between these two diseases: (i) the presence of Onchocerca volvulus in the central nervous system; (ii) the pathogenicity of various O. volvulus strains; (iii) immunological mechanisms involving cross-reactive immunization or cytokine production during infection; and (iv) the triggering role of insomnia due to itching. PMID- 16307907 TI - Can mosquitoes help to unravel the community structure of Plasmodium species? AB - There has been a recent revival in attempts to understand changes in patterns of abundance of Plasmodium spp. that infect humans. This has been driven by the purportedly beneficial effects of co-infection on clinical pathology and the recognition of Plasmodium vivax as a public health problem in its own right. In contrast to the attention given to mixed-species infections in humans, parasite infections and interactions within the mosquito vector remain poorly documented, even though the distribution of vector-borne parasites such as Plasmodium spp. depends on vector-vertebrate and, crucially, vector-parasite interactions. To understand malaria epidemiology and to design appropriate control measures, this gap must be re-addressed. PMID- 16307908 TI - T-cell receptor repertoire of circulating gamma delta T-cells in Takayasu's arteritis. AB - We studied T-cell receptor (TCR) repertoire of circulating gamma delta (gammadelta) T-cells in 20 patients with Takayasu's arteritis (TA), 20 healthy controls (HC), 7 follow up TA patients, and 10 patients with rheumatoid arthritis (RA) and 5 Wegener's granulomatosis (WG) patients as disease controls. Patients with TA (8.1 +/- 5.1%) compared to HC (3.7 +/- 2.1%, P = 0.014), RA (4.8 +/- 0.6%, P = 0.032), and WG (4.2 +/- 0.8%, P = 0.030) as well as active TA compared to inactive TA (13.9 +/- 4.1% vs. 4.9 +/- 1.5%; P < 0.001) had higher number of gammadelta T-cells. The numbers of Vdelta1+ cells were significantly higher in patients with TA (40.0 +/- 20.8%) than HC (13.1 +/- 8.0%; P = 0.001), RA (19.5 +/ 1.8%, P = 0.004), and WG (17.0 +/- 3.9%, P = 0.007). The numbers of gammadelta T cells normalized in all the 7 patients after 180 days of follow up (13.9 +/- 4.1% vs. 6.9 +/- 2.5%; P = 0.001). We also observed higher number of activated and IFN gamma producing gammadelta T-cells in active TA. Our data show that gammadelta T cells particularly those bearing Vdelta1 TCR may have an important role in the immunopathogenesis of TA. PMID- 16307909 TI - DNA-based electrochemical biosensors for monitoring of bis-indoles as potential antitumoral agents, chemistry, X-ray crystallography. AB - Facile and practical electrochemical DNA bioassay, X-ray diffraction analysis, synthesis and 1H and 13C NMR data of the 5,5'-disubstituted-3,3'-methanediyl-bis indoles are reported. On the basis of electrochemical measurements we have hypothesized that the analyzed bis-indoles have an effect on human tumor cells due to DNA binding at adenine-thymidine deoxynucleotides rich region in a concentration/substituent dependent manner. Interesting N-H...pi and hydrogen bonding intermolecular interactions were observed which may differentiate their biological features. The 5,5'-dimethoxy-3,3'-methanediyl-bis-indole (2) was found to reduce considerably the growth of cancer cell lines HOP-92 (lung), A498 (renal) and MDA-MB-231/1TCC (breast). The results indicate that title compounds could be interesting as potential antitumoral chemotherapeutics. PMID- 16307910 TI - Development of an on-line extraction turbulent flow chromatography tandem mass spectrometry method for cassette analysis of Caco-2 cell based bi-directional assay samples. AB - Caco-2 cells are frequently used for screening compounds for their permeability characteristics and P-glycoprotein (P-gp) interaction potential. Bi-directional permeability studies performed on Caco-2 cells followed by analysis by HPLC-UV or LC-MS method constitutes the "method of choice" for the functional assessment of efflux characteristics of a test compound. A high throughput LC-MS/MS method has been developed using on-line extraction turbulent flow chromatography coupled to tandem mass spectrometric detection to analyze multiple compounds present in Hanks balanced salt solution in a single analytical run. All standard curves (P gp substrates: quinidine, etoposide, rhodamine 123, dexamethasone, and verapamil and non-substrates: metoprolol, sulfasalazine, propranolol, nadolol, and furosemide) were prepared in a cassette mode (ten-in-one) while Caco-2 cell incubations were performed both in discreet mode and in cassette mode. The standard curve range for most compounds was 10-2500 nM with regression coefficients (R(2)) greater than 0.99 for all compounds. The applicability and reliability of the analysis method was evaluated by successful demonstration of efflux ratio greater than 1 for the P-gp substrates studied in the Caco-2 cell model. The use of cassette mode analysis through selected reaction monitoring mass spectrometry presents an attractive option to increase the throughput, sensitivity, selectivity, and efficiency of the model over discreet mode UV detection. PMID- 16307911 TI - An essential connection: link between Hsp70's domains at last. AB - Communication between the ATPase and substrate binding domains of Hsp70 is critical for regulated interaction between this molecular chaperone and its client proteins. In this issue of Molecular Cell, Jiang et al. (2005) report the structure of an intact Hsp70, revealing critical interactions between the two domains. PMID- 16307912 TI - SUMO changes Sox for developmental diversity. AB - The finding that posttranslational modification of the SoxE transcription factors by SUMO regulates specific developmental programs (Taylor and LaBonne [2005], in the journal Developmental Cell) highlights the biological significance of SUMOylation in gene expression and underscores how much there is yet to learn about the function and regulation of this modification. PMID- 16307913 TI - Regulation of rRNA processing: a role for a unique GTPase. AB - Recent findings by Karbstein et al. (2005 [this issue of Molecular Cell]) reveal that association of the preribosomal biosynthesis factors U3 snoRNA and Rcl1p is controlled by the GTPase Bms1p, suggesting that regulatory events are involved in the formation of ribosome biogenesis complexes. PMID- 16307914 TI - Dephosphorylation shows SR proteins the way out. AB - To address the role of the RS domain in shuttling and how it is differentially required for constitutive and alternative splicing, Lin et al. (2005 [in the November 11 issue of Molecular Cell]) employ an elegant somatic complementation system to reveal a novel phosphorylation-dependent mechanism regulating distinct recycling pathways for SR proteins during mRNP maturation. PMID- 16307915 TI - Persistent glycoprotein misfolding activates the glucosidase II/UGT1-driven calnexin cycle to delay aggregation and loss of folding competence. AB - The UDP-glucose:glycoprotein glucosyltransferase (UGT) is a central player of glycoprotein quality control in the endoplasmic reticulum (ER). UGT reglucosylation of nonnative glycopolypeptides prevents their release from the calnexin cycle and secretion. Here, we compared the fate of a glycoprotein with a reversible, temperature-dependent folding defect in cells with and without UGT1. Upon persistent misfolding, tsO45 G was slowly released from calnexin and entered a second level of retention-based ER quality control by forming BiP/GRP78 associated disulfide-bonded aggregates. This correlated with loss in the ability to correct misfolding. Deletion of UGT1 did not affect the stringency of ER quality control. Rather, it accelerated release from calnexin and transfer to the second ER quality control level, but it did so after an unexpectedly long lag, showing that cycling in the calnexin chaperone system is not frenetic, as claimed by existing models, and is fully activated only upon persistent glycoprotein misfolding. PMID- 16307916 TI - Structural basis of interdomain communication in the Hsc70 chaperone. AB - Hsp70 family proteins are highly conserved chaperones involved in protein folding, degradation, targeting and translocation, and protein complex remodeling. They are comprised of an N-terminal nucleotide binding domain (NBD) and a C-terminal protein substrate binding domain (SBD). ATP binding to the NBD alters SBD conformation and substrate binding kinetics, but an understanding of the mechanism of interdomain communication has been hampered by the lack of a crystal structure of an intact chaperone. We report here the 2.6 angstroms structure of a functionally intact bovine Hsc70 (bHsc70) and a mutational analysis of the observed interdomain interface and the immediately adjacent interdomain linker. This analysis identifies interdomain interactions critical for chaperone function and supports an allosteric mechanism in which the interdomain linker invades and disrupts the interdomain interface when ATP binds. PMID- 16307917 TI - Chaperoned ubiquitylation--crystal structures of the CHIP U box E3 ubiquitin ligase and a CHIP-Ubc13-Uev1a complex. AB - CHIP is a dimeric U box E3 ubiquitin ligase that binds Hsp90 and/or Hsp70 via its TPR-domain, facilitating ubiquitylation of chaperone bound client proteins. We have determined the crystal structure of CHIP bound to an Hsp90 C-terminal decapeptide. The structure explains how CHIP associates with either chaperone type and reveals an unusual asymmetric homodimer in which the protomers adopt radically different conformations. Additionally, we identified CHIP as a functional partner of Ubc13-Uev1a in formation of Lys63-linked polyubiquitin chains, extending CHIP's roles into ubiquitin regulation as well as targeted destruction. The structure of Ubc13-Uev1a bound to the CHIP U box domain defines the basis for selective cooperation of CHIP with specific ubiquitin-conjugating enzymes. Remarkably, the asymmetric arrangement of the TPR domains in the CHIP dimer occludes one Ubc binding site, so that CHIP operates with half-of-sites activity, providing an elegant means for coupling a dimeric chaperone to a single ubiquitylation system. PMID- 16307918 TI - Akt blocks breast cancer cell motility and invasion through the transcription factor NFAT. AB - The phosphoinositide 3-kinase (PI 3-K) signaling axis is intimately associated with deregulated cancer cell growth, primarily by promoting increased survival through Akt/PKB (protein kinase B). However, there is relatively little information on the role of Akt in cancer cell motility, a key phenotype of invasive carcinomas. Here we report that activation of Akt inhibits carcinoma migration and invasion of breast cancer cells. Conversely, downregulation of Akt using RNA interference increased migration and invasion. Akt blunts invasion by inhibiting the transcriptional activity of NFAT (nuclear factor of activated T cells). Specifically, signaling through Akt reduces NFAT expression levels due to ubiquitination and proteasomal degradation, mediated by the E3 ubiquitin ligase HDM2. These results indicate that while Akt can promote tumor progression through increased cell survival mechanisms, it can block breast cancer cell motility and invasion by a mechanism that depends, at least in part, on the NFAT transcription factor. PMID- 16307919 TI - Functional human telomeres are recognized as DNA damage in G2 of the cell cycle. AB - Telomeres have to be distinguished from DNA breaks that initiate a DNA damage response. Proteins involved in the DNA damage response have previously been found at telomeres in transformed cells; however, the importance of these factors for telomere function has not been understood. Here, we show that telomeres of telomerase-negative primary cells recruit Mre11, phosphorylated NBS1, and ATM in every G2 phase of the cell cycle. This recruitment correlates with a partial release of telomeric POT1; moreover, telomeres were found to be accessible to modifying enzymes at this time in the cell cycle, suggesting that they are unprotected. Degradation of the MRN complex, as well as inhibition of ATM, led to telomere dysfunction. Consequentially, we propose that a localized DNA damage response at telomeres after replication is essential for recruiting the processing machinery that promotes formation of a chromosome end protection complex. PMID- 16307920 TI - Crossover and noncrossover pathways in mouse meiosis. AB - During meiosis, recombination between homologous chromosomes generates crossover (CR) and noncrossover (NCR) products. CRs establish connections between homologs, whereas intermediates leading to NCRs have been proposed to participate in homologous pairing. How these events are differentiated and regulated remains to be determined. We have developed a strategy to detect, quantify, and map NCRs in parallel to CRs, at the Psmb9 meiotic recombination hot spot, in male and female mouse germ lines. Our results report direct molecular evidence for distinct CR and NCR pathways of DNA double-strand break (DSB) repair in mouse meiosis based on three observations: both CRs and NCRs require Spo11, NCR products have shorter conversion tracts than CRs, and only CRs require the MutL homolog Mlh1. We show that both products are formed from middle to late pachytene of meiotic prophase and provide evidence for an Mlh1-independent CR pathway, where mismatch repair does not require Mlh1. PMID- 16307921 TI - Progressive activation of DNA replication initiation in large domains of the immunoglobulin heavy chain locus during B cell development. AB - In mammalian cells, the replication of tissue-specific gene loci is believed to be under developmental control. Here, we provide direct evidence of the existence of developmentally regulated origins of replication in both cell lines and primary cells. By using single-molecule analysis of replicated DNA (SMARD), we identified various groups of coregulated origins that are activated within the Igh locus. These origin clusters can span hundreds of kilobases and are activated sequentially during B cell development, concomitantly with developmentally regulated changes in chromatin structure and transcriptional activity. Finally, we show that the changes in DNA replication initiation that take place during B cell development, within the D-J-C-3'RR region, occur on both alleles (expressed and nonexpressed). PMID- 16307922 TI - The Bur1/Bur2 complex is required for histone H2B monoubiquitination by Rad6/Bre1 and histone methylation by COMPASS. AB - To date, several classes of enzymes have been shown to affect transcription by catalyzing the modifications of nucleosomes via methylation. Employing our global proteomic screen, GPS, we have determined that the loss of Bur2, a component of the Bur1/Bur2 cyclin-dependent protein kinase, results in a decrease in histone H3(K4) methylation catalyzed by COMPASS. Furthermore, Bur1/Bur2 is required for histone H2B monoubiquitination by Rad6/Bre1. The effect on histone monoubiquitination and methylation is the result of defective Bur1/Bur2-mediated phosphorylation of Rad6 on its serine residue 120 and proper recruitment of the Paf1 complex to chromatin. We have also demonstrated that serine 120 of Rad6 is required for histone H2B monoubiquitination and the regulation of gene expression in vivo. Our results identify in vivo substrates for Bur1/Bur2, thus linking its role to transcriptional elongation and demonstrating a potential activation mechanism for histone H2B monoubiquitination by the Rad6/Bre1 complex. PMID- 16307923 TI - Monoubiquitination of human histone H2B: the factors involved and their roles in HOX gene regulation. AB - In yeast, histone H2B monoubiquitination is a cotranscriptional event regulating histone H3 methylation at lysines 4 and 79. However, mammalian H2B monoubiquitination remains poorly understood. We report that in humans, the 600 kDa RNF20/40 complex is the E3 ligase and UbcH6 is the ubiquitin E2-conjugating enzyme for H2B-Lys120 monoubiquitination. RNF20 and RNF40 are both homologs of Bre1, the E3 ligase in the yeast case. UbcH6 physically interacts with RNF20/40 and with the hPAF complex. Formation of a trimeric complex with hPAF stimulates H2B monoubiquitination activity in vitro. Accordingly, UbcH6, RNF20/40, and the hPAF complex are recruited to transcriptionally active genes in vivo. RNF20 overexpression leads to elevated H2B monoubiquitination, subsequently higher levels of methylation at H3 lysines 4 and 79, and stimulation of HOX gene expression. In contrast, RNAi against the RNF20/40 complex or hPAF complex reduces H2B monoubiquitination, lowers methylation levels at H3 lysines 4 and 79, and represses HOX gene expression. PMID- 16307924 TI - Destabilization of the P site codon-anticodon helix results from movement of tRNA into the P/E hybrid state within the ribosome. AB - Retention of the reading frame in ribosomal complexes after single-round translocation depends on the acylation state of the tRNA. When tRNA lacking a peptidyl group is translocated to the P site, the mRNA slips to allow re-pairing of the tRNA with a nearby out-of-frame codon. Here, we show that this ribosomal activity results from movement of tRNA into the P/E hybrid state. Slippage of mRNA is suppressed by 3' truncation of the translocated tRNA, increased MgCl2 concentration, and mutation C2394A of the 50S E site, and each of these conditions inhibits P/E-state formation. Mutation G2252U of the 50S P site stimulates mRNA slippage, suggesting that decreased affinity of tRNA for the P/P state also destabilizes mRNA in the complex. The effects of G2252U are suppressed by C2394A, further implicating the P/E state in mRNA destabilization. This work uncovers a functional attribute of the P/E state crucial for understanding translation. PMID- 16307925 TI - Involvement of 16S rRNA nucleotides G1338 and A1339 in discrimination of initiator tRNA. AB - Three consecutive G-C pairs in the anticodon stem are a key discriminatory feature of initiator tRNA and are required for its selection by IF3. Here, we have mutated two 16S rRNA nucleotides, G1338 and A1339, which provide the sole contact to the G-C pairs of tRNA(fMet) bound to the ribosomal P site. We have tested their effects on translational activities in vivo and have affinity purified mutant 30S subunits for functional analysis in vitro. Our results are consistent with the formation of Type II and I minor interactions, respectively, between G1338 and A1339 and the anticodon stem of tRNA and suggest that these interactions play a role in tRNA(fMet) discrimination by IF3. Moreover, our findings indicate that discrimination also involves recognition of at least one additional feature of the tRNA(fMet) anticodon stem loop. PMID- 16307926 TI - An essential GTPase promotes assembly of preribosomal RNA processing complexes. AB - Ribosome biogenesis in eukaryotes is a highly regulated process involving hundreds of transiently associated proteins and RNAs. Although most of these assembly factors have been genetically linked to specific step(s) in the biogenesis pathway, their biochemical functions are generally unknown. Bms1, an essential protein in yeast, is the only known GTPase required for biosynthesis of the 40S ribosomal subunit and interacts with Rcl1, an essential protein suggested to be an endonuclease. Here, we show thermodynamic coupling in the binding of Bms1 to GTP, Rcl1, and U3 small nucleolar RNA (snoRNA), an essential RNA that base pairs to pre-rRNA. Rcl1 binding to preribosomes is severely limited in yeast cells expressing a Bms1 mutant defective for Rcl1 binding. Additionally, we provide evidence that the C-terminal domain of Bms1 acts as an intramolecular GTPase-activating protein. Together, these data suggest that Bms1 functions as a GTP-regulated switch to deliver Rcl1 to preribosomes, providing molecular insight into preribosome assembly. PMID- 16307927 TI - The DEAD-box protein Dbp5p is required to dissociate Mex67p from exported mRNPs at the nuclear rim. AB - Eukaryotic mRNAs are exported from the nucleus to the cytoplasm as complex mRNA protein particles (mRNPs), and translocation through the nuclear pore complex (NPC) is accompanied by extensive structural changes of the mRNP. We have tested the hypothesis that the DEAD-box ATPase Dbp5p is required for such an mRNP rearrangement. In dbp5 mutant cells, the mRNA export receptor Mex67p accumulates on mRNA. This aberrant accumulation of Mex67p with RNA and the cold-sensitive growth phenotype of a dbp5 allele are suppressed by a mex67 mutation. Moreover, Mex67 bound mRNA accumulates at the nuclear rim in a temperature-sensitive dbp5 mutant when the nuclear exosome is impaired. Importantly, although accumulation of Mex67p-containing mRNPs is also observed when a nuclear basket component is mutated, these mRNPs still contain the nuclear export factor Yra1p. In contrast, the dbp5-trapped mRNPs lack Yra1p. We propose that Dbp5p's function is specifically required to displace Mex67p from exported mRNPs, thus terminating export. PMID- 16307928 TI - Visceral injuries in nonaccidental trauma: spectrum of injury and outcomes. AB - BACKGROUND: Nonaccidental trauma (NAT) causes significant morbidity and mortality in children. The purpose of this study was to characterize visceral injuries associated with NAT and the management and outcomes of children with these injuries. METHODS: During a 7-year period, children admitted to our regional pediatric trauma center with a diagnosis of NAT were identified and their injuries characterized. RESULTS: NAT accounted for 7% (n = 265 of 3705) of all trauma admissions during the period of study. Visceral injuries were diagnosed in 9% (n = 24 of 265) of NAT patients. Compared with the remaining NAT population, children with visceral injuries were similar in age and sex but had higher injury severity scores (21 vs. 17, P < .05). There was a high coincidence of thoracic trauma and nonburn integumental injuries in abdominally injured NAT patients. Children with visceral injuries were more likely to undergo emergent operations (46% [11 of 24] vs. 5% [15 of 241], P < .0001) than those without. However, there was no difference in Intensive Care Unit stay, hospital stay, or overall mortality for children with visceral injuries compared with those without. CONCLUSIONS: Visceral injuries are not uncommon in NAT, and these injuries often require emergent operative intervention. Thus, prompt evaluation and treatment by a surgeon remains a critical step in the management of children with NAT. PMID- 16307929 TI - The outcome of open pelvic fractures in the modern era. AB - BACKGROUND: Recent series have reported that the mortality rate of open pelvic fractures has decreased to < 10%. These injuries are often associated with intra abdominal visceral damage, although few series have documented the prognostic significance of this injury complex. METHODS: A retrospective review in an urban level I trauma center of all patients who sustained open pelvic fracture between 1995 and 2004. RESULTS: Forty-four patients were identified as having sustained open pelvic fracture. Average Injury Severity Score was 30, with 77% of patients having a score > or = 16. Overall mortality was 45% (n = 20): 11 early deaths and 9 late deaths at an average of 17 days. Vertical shear injuries, although rare, were universally fatal. Other risk factors for overall mortality included revised trauma score, Injury Severity Score, transfusion requirement, Faringer zones I or II injury, Gustilo grade III soft tissue injury, need for therapeutic angiography, and presence of intra-abdominal injury, the latter of which conferred 89% mortality. Risk factors for late deaths also included pelvic sepsis, which occurred in 5 patients and was fatal in 3 (60%). CONCLUSIONS: The morbidity of open pelvic fractures remains high. Associated intra-abdominal injury or active arterial bleeding requiring therapeutic angiography is associated with a grim prognosis. There is a continuing need for new therapeutic approaches to this injury complex. PMID- 16307930 TI - Patterns of injury and functional outcome after hanging: analysis of the National Trauma Data Bank. AB - PURPOSE: Hanging has become the second most common method of attempted suicide among adolescents, but there is little relevant epidemiologic or outcome data in the trauma literature. Additionally, there are no studies examining the degree of functional disability among survivors of hanging injury. METHODS: The National Trauma Data Bank was queried for all patients with an E-code diagnosis of hanging injury. Demographic and injury pattern data were analyzed. Disability at discharge was assessed using the functional independence measure (FIM) scores for feeding, locomotion, and expression (range 1 = full disability to 4 = no disability). Univariate and multivariate analysis was performed to identify independent predictors of mortality and degree of functional disability at discharge. RESULTS: There were 655 patients identified (84% male) with a mean age of 30.3 years and mean injury severity score (ISS) of 9. There were 92 (14%) deaths in the emergency department (ED) and 119 (18%) deaths after admission, for an overall mortality rate of 33%. Excluding ED deaths, survivors had significantly higher Glasgow coma scores (GCS) at the scene (8 vs. 4) and in the ED (9 vs. 3), a lower ED base deficit (4 vs. 9), and lower ISS (6 vs. 15, all P < .01) compared with nonsurvivors. The strongest independent predictor of hospital mortality was ED GCS <15 (odds ratio 16.1, P < .01); the mortality rate was 1.5% for patients with an ED GCS of 15 versus 29% for any GCS <15. Of patients who survived to discharge (n = 277), 84% were functionally independent (total FIM = 12), and 10% had severe functional disabilities in feeding, expression, or locomotion (FIM <3). Patients with severe disability had a higher incidence of intracranial (38% vs. 19%) and chest injury (19% vs. 5%) but surprisingly demonstrated equivalent rates of vascular (0% vs. 2.6%) and spinal injury (11% vs. 12%) compared with those without severe disability. Independent predictors of functional outcome were ISS and ED GCS (both P < .01). There was no severe functional disability at discharge among patients with an ED GCS of 15 compared with a 15% severe disability rate if the ED GCS was <15. CONCLUSIONS: Hanging injuries are associated with a high overall mortality rate, with the admission GCS being the best independent predictor of outcome. However, the majority of survivors have little to no functional disability. The presence of severe disability at discharge is mainly attributed to intracranial and thoracic injury. PMID- 16307931 TI - Occult traumatic hemothorax: when can sleeping dogs lie? AB - BACKGROUND: Size of traumatic occult hemothorax on admission requiring drainage has not been defined. Computed axial tomography (CAT) may guide drainage criteria. METHODS: A retrospective review of patients with hemothoraces on CAT was performed. Extrapolating previously described methods of pleural fluid measurement, hemothoraces were quantified using the fluid stripe in the dependent pleural "gutter." Data included patient age, injury severity, and intervention (thoracentesis or tube thoracostomy). RESULTS: Seventy-eight patients with 99 occult hemothoraces met the criteria for study inclusion: 52 hemothoraces qualified as "minimal" and 47 as "moderate/large." Eight patients (15%) in the minimal group and 31 patients (66%) in the moderate/large group underwent intervention (P < .001). There was no difference in patient age, injury severity, ventilator requirement, or presence of pulmonary contusion. CONCLUSIONS: CAT in stable blunt-trauma patients can predict which patients with occult hemothorax are likely to undergo intervention. Patients with hemothorax > or = 1.5 cm on CAT were 4 times more likely to undergo drainage intervention compared with those having hemothorax < 1.5 cm. PMID- 16307932 TI - Screening for blunt cerebrovascular injuries is cost-effective. AB - BACKGROUND: Recent reports have argued that screening for blunt carotid injury is futile and have called for a cost analysis. Our data previously supported screening asymptomatic trauma patients for blunt cerebrovascular injury (BCVI) to prevent associated neurologic sequelae. Our hypothesis is that aggressive angiographic screening for BCVI based on a patient's injury pattern and symptoms allows for early diagnosis and treatment and is cost-effective because it prevents ischemic neurological events (INEs). METHODS: Beginning in January 1996, we began comprehensive screening using 4-vessel cerebrovascular angiography based on injury patterns; these patients have been followed-up prospectively. Patients without contraindications received antithrombotic therapy immediately for documented BCVI. RESULTS: From January 1996 through June 2004, there were 15,767 blunt-trauma patient admissions to our state-designated level I urban trauma center, of which 727 patients underwent screening angiography. Twenty-one patients presented with signs or symptoms of neurologic ischemia before diagnosis. BCVI was identified in 244 patients (34% screening yield); the majority were men (68%) with a mean age of 35 +/- 3.7 years and mean Injury Severity Score of 28 +/- 3.8. Asymptomatic patients (n = 187) were treated (heparin in 117, low molecular-weight heparin in 11, and antiplatelet in 59); 1 patient had a stroke (0.5%). Using estimated stroke rate by grade of injury, we averted neurologic events in 32 asymptomatic patients with antithrombotic treatment. Of the 48 asymptomatic patients who did not receive adequate anticoagulation, 10 (21%) had an INE. Patients with BCVI-related neurologic events had a statistically higher percentage requiring discharge to rehabilitation facilities (50% vs. 77% for carotid artery injury [CAI]), a higher percentage requiring rehabilitation for BCVI-related stroke (0% vs. 55% for CAI), and a higher stroke-related mortality rate (0% vs. 21% for CAI and 0% vs. 17% for vertebral artery injury) than those without neurologic events. CONCLUSIONS: The cost of long-term rehabilitation care and human life after BCVI-associated neurologic events is substantial. Surgeons caring for the multiply injured should screen for carotid and vertebral artery injuries in high-risk patients. PMID- 16307933 TI - Carotid artery stenting with neuroprotection: assessing the learning curve and treatment outcome. AB - PURPOSE: Carotid artery stenting (CAS) has emerged as an acceptable treatment alternative in high-risk patients with carotid stenosis. The purpose of this study was to assess the effect of the learning curve on treatment complications and the clinical outcomes of CAS. METHODS: Clinical variables and treatment outcomes of 200 consecutive CAS procedures in 182 patients (mean age 72 years) with carotid stenosis > or = 70% during a 40-month period were analyzed. Four sequential groups (groups I, II, III, and IV) of 50 consecutive interventions were compared with regard to technical success, periprocedural complications, and treatment outcomes. RESULTS: Treatment indications and relevant risk factors were similar among the 4 groups. The overall technical success and combined 30-day stroke and death rates were 98% and 2.5%, respectively. An increase in the technical success rate was noted in the latter 3 groups compared with group I (P < .05). Total procedural time and contrast volume were significantly higher in group I compared with the latter 3 groups (P < .05). The intraoperative anticoagulation regimen was changed from intravenous heparin combination to bivalirudin after the first 54 patients, which resulted in decreased bleeding complications in groups III and IV (P = 0.03) compared with the first group. The 30-day stroke and death rate in groups I and II were 8% and 2%, respectively, and was decreased significantly in groups III and IV (0% and 0%, respectively, P < .05). A Cox regression model identified procedural volume (P = .03) as a predictor of decreased complication rate. CONCLUSIONS: CAS with neuroprotection can provide excellent treatment outcomes. Our experience demonstrates a procedure associated learning curve as evidenced by decreased procedure-related complications, fluoroscopic time, and contrast volume occurring with increased physician experience. Procedural success was also enhanced partly by endovascular device refinement and an improved anticoagulation regimen. Successful CAS outcomes can be achieved once physicians overcome the initial procedure-related learning curve. PMID- 16307934 TI - Are temporary inferior vena cava filters really temporary? AB - BACKGROUND: Despite significant risk for venous thromboembolism, severely injured trauma patients often are not candidates for prophylaxis or treatment with anticoagulation. Long-term inferior vena cava (IVC) filters are associated with increased risk of postphlebitic syndrome. Retrievable IVC filters potentially offer a better solution, but only if the filter is removed; our hypothesis is that the most of them are not. METHODS: This retrospective study queried a level I trauma registry for IVC filter insertion from September 1997 through June 2004. RESULTS: One IVC filter was placed before the availability of retrievable filters in 2001. Since 2001, 27 filters have been placed, indicating a change in practice patterns. Filters were placed for prophylaxis (n = 11) or for therapy in patients with pulmonary embolism or deep vein thrombosis (n = 17). Of 23 temporary filters, only 8 (35%) were removed. CONCLUSIONS: Surgeons must critically evaluate indications for IVC filter insertion, develop standard criteria for placement, and implement protocols to ensure timely removal of temporary IVC filters. PMID- 16307935 TI - Catheter-directed thrombolysis and/or thrombectomy with selective endovascular stenting as alternatives to systemic anticoagulation for treatment of acute deep vein thrombosis. AB - PURPOSE: To evaluate thrombolysis and/or thrombectomy and selective endovascular stent placement in treating acute deep vein thrombosis (DVT). METHODS: During a 5 year period, 28 patients were treated with catheter-directed thrombolytics and/or thrombectomy with endovascular stent placement. Seventy-two percent (n = 20) of patients had symptoms for less < or = 14 days; 14% (n = 4) had symptoms for > 14 days. Fourteen percent (n = 4) had recurrent symptoms; 43% (n = 12) had ileofemoral DVT, and 57% (n = 16) had common femoral, superficial femoral, and/or popliteal DVT; and 11% (n = 3) had thrombus extending into the inferior vena cava. RESULTS: Eighteen percent (n = 5) of patients had complete thrombolysis of the thrombus; 72% (n = 20) had partial thrombolysis. Twenty-two stents were also placed in 12 patients. Average follow-up was 15.5 months; 80% had long-term patency. CONCLUSIONS: Catheter-directed thrombolysis and/or thrombectomy and selective stent placement are effective alternatives to systemic anticoagulation in the treatment of DVT. More studies are needed to determine specific indications and to validate long-term efficacy. PMID- 16307936 TI - The difficult hemodialysis access extremity: proximal radial arteriovenous fistulas and the role of angioscopy and valvulotomes. AB - BACKGROUND: Native arteriovenous (AV) fistulas (NAVF) offer significantly lower risks than grafts or catheters. Individuals with a difficult access extremity (DAE) are often viewed as unsuitable for NAVFs. The proximal radial artery (PRA) NAVF offers a safe and reliable opportunity for a direct fistula in most patients, and we find it an important surgical option in the DAE. METHODS: Consecutive vascular access operations were reviewed to find individuals with DAE. We defined the DAE patient group as those individuals where a wrist (Cimino) or upper arm brachiocephalic NAVF was not possible or was predicted to fail. RESULTS: Preoperative physical and ultrasound examinations identified 58 individuals with DAE. Mean age was 56 years (range 11 to 87), 34 were female, 29 were diabetic, and 27 had previous access surgery. NAVFs were constructed in all patients. No grafts were utilized. Forty-six patients had a PRA NAVF constructed. NAVF patency was 91%. Twenty-three patients required retrograde angioscopy or passage of a valvulotome to gain forearm access. Twenty-one of these 23 individuals maintained an open NAVF segment in the forearm. CONCLUSION: NAVFs were constructed in all patients. PRA NAVFs play an important role in extending hemodialysis by NAVF for this difficult patient group. Forearm access is often possible in these patients and may be successfully augmented by angioscopy or valvulotomes. PMID- 16307937 TI - Long-term results of laparoscopic Heller myotomy with partial fundoplication for the treatment of achalasia. AB - BACKGROUND: Treatment options for achalasia include medications, endoscopic balloon dilation, injection of botulinum toxin, or surgery. METHODS: The clinical course of 75 consecutive patients who underwent minimally invasive Heller myotomy and partial fundoplication for achalasia between 1991 and 2001 was reviewed by means of a questionnaire. RESULTS: Mean follow-up was 5.3 (range .8 to 10.9) years. Sixty-four percent of questionnaires were returned. Thirty-seven patients (84%) felt much better and 6 (14%) slightly better; 1 (2%) rated the result as unchanged. Twenty-six patients (59%) experienced weight gain. Seven patients (16%) had persistent swallowing problems and 5 (11%) reported frequent reflux. Twenty-five percent underwent additional therapy, including dilation (n = 8, 18%), repeat surgery (n = 2, 5%), and botulinum toxin injection (n = 2, 5%). Eighteen patients (41%) were using a proton pump inhibitor or H2 blocker, three were on a calcium channel blocker (7%), and 1 was using nitroglycerine (2%). CONCLUSION: Laparoscopic Heller myotomy can achieve short- and long-term results comparable to open surgery and should be considered the treatment of choice for patients suffering from achalasia. Despite the frequent need for further therapy, patient satisfaction is good. PMID- 16307938 TI - Laparoscopic converted to open cholecystectomy minimally prolongs hospitalization. AB - BACKGROUND: There has been little information about the length of stay (LOS) after laparoscopic converted to open cholecystectomy (CON) in the past few years. The aim of this study was to evaluate the LOS and postoperative complications for elective CON in a more recent time period. METHODS: A retrospective chart review was performed of all patients admitted to the Day Surgery Unit for elective laparoscopic cholecystectomy (LC) converted to open cholecystectomy (OC) from January 2000 through December 2003. Indications for CON, operative time, LOS, pain control, and complications were evaluated. RESULTS: The CON rate was 3%, and the reason for CON to open was most commonly cited as inability to identify anatomy. On average, patients were discharged on postoperative day 3 (range 2 to 8). The postoperative complication rate was 17%. CONCLUSIONS: When the dissection is tedious, the surgeon should feel comfortable in converting from laparoscopic to open cholecystectomy. This can be done with the knowledge that it does not add significant length of stay as previously reported. PMID- 16307939 TI - Laparoscopic colorectal surgery in the complicated patient. AB - BACKGROUND: Major comorbidities are recognized risk factors in colorectal surgery. We examine here the feasibility and safety of laparoscopic colorectal surgery (LC) in the complicated, high-risk patient. METHODS: From July 2003 to October 2004, 107 consecutive patients undergoing LC were prospectively studied. Complicated patients were defined as age >80 years, body mass index (BMI) >30, and/or American Society of Anesthesiology level III or IV. A group of case matched controls undergoing open surgery (OC) during a similar time period were retrospectively reviewed. The 2 groups were compared and assessed for major and minor morbidity and mortality. RESULTS: Overall morbidity was higher in the OC group 52% versus 26%. Minor complications compared at 31% OC versus 9% LC and major at 21% and 17%, respectively. With LC, advancement to discharge was more rapid and discharge home more likely than to a care facility. CONCLUSION: With proper patient selection and laparoscopic experience, LC can be performed in the complicated patient without undue morbidity and mortality. PMID- 16307940 TI - The utility of capsule endoscopy and its role for diagnosing pathology in the gastrointestinal tract. AB - BACKGROUND: Capsule endoscopy (CE) is a new device that enables visualization of areas of the small bowel that were previously inaccessible through other noninvasive procedures. The purpose of this study is to evaluate this new diagnostic tool and its efficacy in finding occult GI tract pathology. METHODS: A single-institution retrospective review was completed on patients undergoing CE from January 2002 to September 2004. Data evaluated included indications for CE, results of previous studies, CE findings, and complications of the CE study. RESULTS: A total of 702 CE studies in 652 patients were performed during the study period. Suspicious GI bleeding presenting as anemia, guaiac positive stools, or history of gross bleeding were the most common reasons to perform CE (75.8%). Other indications included abdominal pain (11.5%), diarrhea (3.1%), or others (9.5%). In studies performed for GI bleeding (N = 532), a source was found in 49.3% of CE studies. Arteriovenous malformation (AVM) was the most common reported finding (43.9%), followed by ulcer (24.1%), colon or gastric pathology (14.1%), mass/tumor (9.1%), and stricture (6.9%). Patients with abdominal pain (n = 81) had findings 46.9% of the time including edema/ulcer (47.4%), stricture (10.5%), mass/tumor (26.3%), gastric pathology (10.5%), AVM (2.6%), or sprue (2.6%). Patients with diarrhea (n = 22) had findings 45.5% of the time including edema/ulcer (75%), mass/tumor (12.5%), or sprue (12.5%). A total of 66 patients underwent operative exploration after a CE study at this institution either because of the observed findings or for other reasons. There were 12 (1.7%) CE studies in which the capsule was retained and required surgical removal. Pathology at the retention site included benign strictures or adhesions (n = 9, 75%), Crohn's stricture (n = 1, 8.3%) carcinoid tumor (n = 1, 8.3%), and villous adenoma (n = 1, 8.3%). CONCLUSIONS: CE is an accurate study to locate abnormalities in the GI tract that may have either been missed by previous diagnostic studies or cannot be observed through other non-invasive means. When used for diagnostic challenges such as GI bleeding with no apparent source, CE can be helpful in guiding surgical decisions in patients and thus should be integrated as part of the diagnostic workup. PMID- 16307941 TI - Cause and treatment of epiphrenic diverticula. AB - BACKGROUND: Epiphrenic diverticula of the esophagus are often associated with a concomitant esophageal motor disorder, which is thought to be the cause of the diverticulum and some of the patient's symptoms. At one time diverticula were best removed via a left thoracotomy, but now the operation can be performed laparoscopically in most cases. We hypothesized that: (1) a motor disorder is the underlying cause of the diverticulum; and (2) optimal treatment consists of laparoscopic resection of the diverticulum, a Heller myotomy, and Dor fundoplication. METHODS: We performed a retrospective review of a prospectively collected database from a university hospital tertiary care center. Between June 1994 and December 2002, we evaluated 21 patients with epiphrenic diverticula. An associated motility disorder of the esophagus was found in 81% of patients (achalasia, 9%; diffuse esophageal spasm, 24%; nonspecific esophageal motility disorder, 24%; nutcracker esophagus, 24%). Seven (33%) of these patients, all with esophageal dysmotility, were referred for treatment. The laparoscopic operation entailed resection of the diverticulum (using an endoscopic stapler), a Heller myotomy, and a Dor fundoplication. RESULTS: All operations were completed laparoscopically. The postoperative course of 6 patients was uneventful and they left the hospital after 72 +/- 21 hours. In 1 patient an acute paraesophageal hernia developed, which was repaired on the second postoperative day. Late follow up (median 57 months) showed that all 7 patients were asymptomatic. CONCLUSIONS: These data support the conclusions that: (1) a primary esophageal motility disorder is the underlying cause of most epiphrenic diverticula; and (2) laparoscopic treatment is successful and should be the method of choice. The diverticular neck can be exposed satisfactorily from the abdomen; a stapler inserted from this angle is better orientated to transect the neck than one inserted through a thoracoscopic approach. Furthermore, the myotomy and fundoplication are much more easily performed from the abdomen than from alternative approaches. PMID- 16307942 TI - Breast cancer in Native American women treated at an urban-based Indian health referral center 1982-2003. AB - BACKGROUND: Breast cancer incidence and survival varies by race and ethnicity. There are limited data regarding breast cancer in Native American women. METHODS: A retrospective chart review was performed of 139 women diagnosed with breast cancer and treated at Phoenix Indian Medical Center in Phoenix, AZ between January 1, 1982 and December 31, 2003. Data points included tribal affiliation, and quantum (percentage American Indian Heritage) along with patient, tumor, and treatment characteristics. RESULTS: Most patients (79%) presented initially with physical symptoms. There were no significant differences based on tribal affiliation; however, higher quantum predicted both larger tumor size and more advanced stage at diagnosis. Obesity also significantly correlated with larger tumor size and more advanced stage. Treatment was inadequate in 21%; this was attributed to traditional beliefs, patient refusal, or financial issues. CONCLUSIONS: When compared to national averages, Native American women presented at a later stage, underutilized screening, and had greater delays to treatment. PMID- 16307943 TI - Surgeon and community factors affecting breast cancer sentinel lymph node biopsy. AB - BACKGROUND: We sought to determine whether the results of sentinel lymph node (SLN) biopsy are related to practice and community factors. METHODS: This prospective study included more than 300 surgeons from a variety of practice environments. Most surgeons had minimal experience with SLN biopsy prior to this study. Patients underwent attempted SLN biopsy, followed by completion axillary dissection. Univariate and multivariate analyses were performed to assess factors related to the SLN identification rate and the false negative rate. RESULTS: A total of 4131 patients were enrolled. SLN identification rate was 93%; the false negative (FN) rate was 7.9%. The only factor that was significantly associated with improved SLN identification rate (odds ratio [OR] 1.60, 95% confidence interval [CI] 1.12 to 2.36, P = .0126) and FN rate (OR 2.39, 95% CI 1.32 to 4.79, P = .0073) was surgeon experience (>20 SLN cases). CONCLUSIONS: Surgeon experience is the major factor that contributes to improved SLN biopsy results. SLN biopsy can be performed equally well by community and academic surgeons. PMID- 16307944 TI - Total skin-sparing mastectomy without preservation of the nipple-areola complex. AB - BACKGROUND: We hypothesized that total skin-sparing mastectomy (TSM) including where the skin overlying the nipple and areola is preserved would be oncologically safe and facilitate improved cosmetic reconstruction. METHODS: A review (May 2003 through January 2005) was completed on all procedures that were performed through an inframammary incision or a previous scar with reconstruction using Botox, AlloDerm, and a subpectoral tissue implant. RESULTS: Thirty-one patients had 50 TSMs. Twelve percent (6/50) of TSMs had the skin of the nipple and areola resected: 4 (14% of tumors) because of tumor involvement and 2 (4%) because of skin necrosis. Fourteen percent of patients had other complications: 4% (2/50) had infection and/or flap necrosis and 10% (5/50) had superficial epidermolysis requiring no intervention, for a total complication rate of 18%. Average cosmetic score was 8.5 (range 4 to 10). No recurrences are evident after mean follow-up of 7.9 +/- 5.4 months. CONCLUSION: Our short-term experience suggests that TSM has an acceptable complication rate, is theoretically oncologically safe, and facilitates an improved cosmetic result. PMID- 16307945 TI - Is incisional biopsy of melanoma harmful? AB - BACKGROUND: In the era of sentinel lymph node (SLN) biopsy, there has been concern that manipulation, injection, and massage of intact primary melanomas (after incisional or shave biopsy) could lead to an artifactual increased rate of SLN micrometastases or an actual increased risk of recurrence. The aim of this study was to evaluate the difference in the incidence of SLN metastasis, locoregional recurrence (LRR), disease-free survival (DFS), distant disease-free survival (DDFS), or overall survival (OS) for patients who undergo excisional versus incisional versus shave biopsy. METHODS: Analysis of database from a multicenter prospective randomized study from centers across the United States and Canada. Eligible patients were 18 to 71 years old, with cutaneous melanoma > or = 1.0 mm Breslow thickness. All patients underwent SLN biopsy using blue dye and radioactive colloid injection. SLNd were evaluated by serial histological sections with S100 immunohistochemistry. Statistical analysis was performed using univariate and multivariate analyses with a significance level of P < .05; survival analysis was performed by the Kaplan-Meier method with the log-rank test. RESULTS: A total of 2,164 patients were evaluated; 382 patients were excluded for lack of biopsy information. Positive SLNs were found in 220 of 1,130 (19.5%), 58 of 281 (20.6%), and 67 of 354 (18.9%) of patients with excisional, incisional, or shave biopsy, respectively (no significant difference). There were significant differences among the 3 biopsy types in ulceration (P = .018, chi2) and regression (P = .022, chi2); there were no differences in age, gender, Breslow thickness, Clark level, lymphovascular invasion, tumor location, or histologic subtype. Biopsy type did not significantly affect LRR, DFS, DDFS, or OS. CONCLUSIONS: The concern that incomplete excision of primary melanomas may result in an increased incidence of SLN micrometastases, artifactual or real, is unfounded. Similarly, there is no evidence that biopsy type adversely affects locoregional or distant recurrence. Although shave biopsy is generally discouraged because it may lead to inaccurate tumor thickness measurements, it does not appear to affect overall patient outcome. PMID- 16307946 TI - Fifteen-year series of skin-sparing mastectomy for stage 0 to 2 breast cancer. AB - BACKGROUND: In 1989, skin-sparing mastectomy started at a number of breast centers in the United States because of an increasing demand for immediate reconstruction and a desire for better cosmetic outcomes. METHODS: To ensure the safety of this new approach, we have reviewed the personal series of a single surgeon using a standardized skin-sparing technique during 1989 to 2004. RESULTS: Skin-sparing mastectomy with immediate reconstruction was performed on 225 patients, and standard mastectomy was performed on 1,022 patients. The age distribution was 8 years younger on average in the reconstructed group. The average follow-up for each group was 49 months. The local recurrence for each group was 1.7% and 1.5% (P > .80). The regional recurrence was 3.8% and 3.9% (P > .80). The average time to local recurrence was similar in each group (33.1 and 32.6 months, P > .80). CONCLUSIONS: A skin-sparing mastectomy does not change the local, regional, or systemic risk to breast cancer patients. PMID- 16307947 TI - Percutaneous tracheostomy: one center's experience with a new modality. AB - BACKGROUND: A retrospective review of our experience with percutaneous tracheostomy was performed to determine our complication rate and pattern of use since this modality was introduced at our institution. METHODS: A retrospective chart review captured all patients in whom tracheostomy was performed or supervised by a trauma/critical care faculty member. Dates of hospital admission, ICU admission, intubation, discontinuation of mechanical ventilation, type and location of procedure, procedural complications, Injury Severity Score, charges, and patient demographics were collected. Percutaneous tracheostomy (PT) and open tracheostomy (OT) experiences were compared. RESULTS: Three hundred sixty-eight tracheostomies were performed (190 OT and 178 PT). The average time to tracheostomy (TTT) for PT patients decreased from 12.7 to 7.4 days. The average TTT for OT patients remained stable at 14.0 days. The complication rate was 3.5%, with 4 complications (1.5%) associated with OT and 9 complications (5.1%) associated with PT. All complications in the PT group occurred before using a single dilator system. The 9 complications in the PT group occurred among 5 surgeons, all before their 11th attempt. PT saves 444 dollars in charges per procedure. CONCLUSION: OT continues to be a safe method of performing tracheostomies. PT has a steep learning curve but can be mastered quickly. Benefits include a shorter time to tracheostomy, elimination of patient transport, and saving in charges. Initial PT attempts should be supervised by an experienced surgeon. PMID- 16307948 TI - Blast lung injury: clinical manifestations, treatment, and outcome. AB - BACKGROUND: Blast lung injury (BLI) is a major cause of morbidity after terrorist bomb attacks (TBAs) and is seen with increasing frequency worldwide. Yet, many surgeons and intensivists have little experience treating BLI. Jerusalem sustained 31 TBAs since 1983, resulting in a local expertise in treating BLI. METHODS: A retrospective study of clinical and radiologic characteristics, management, and outcome of victims of TBAs sustaining BLI who were admitted to ICU during December 1983 to February 2004. Long-term outcome was determined by a telephone interview. RESULTS: Twenty-nine patients met inclusion criteria. Hypoxia and pulmonary infiltrates in chest x-ray were sine qua non for the diagnosis. Seventy-six percent required mechanical ventilation, all within 2 hours of admission. One patient died. Seventy-six percent had no long-term sequelae. CONCLUSIONS: Most patients with significant BLI injury require mechanical ventilation. Late deterioration is rare. Death because of BLI in patients who survived the explosion is unusual. Timely diagnosis and correct treatment result in excellent outcome. PMID- 16307949 TI - Economic modeling comparing trauma and general surgery reimbursement. AB - BACKGROUND: The viability of trauma care as a surgical subspecialty is continually challenged by economic pressures related to reimbursement and opportunity costs. METHODS: The literature was examined for articles focused on economic implications of a trauma focused surgical practice. Economic forecasting techniques were applied using a recalculating spreadsheet to examine charge and revenue generation comparing the effects of numerous variables affecting a trauma or general surgical service. RESULTS: Elective general surgery practices derive the majority of revenues from procedural services, whereas trauma practices derive the majority of revenues from evaluation and management. Only centers with high admission volume can expect trauma surgeons to cover salary and expenses, predictably in association with high opportunity costs. CONCLUSION: The differences in time, effort, and patient volume required for a trauma surgeon to generate revenues comparable to an elective practice are dramatic. The current system creates disincentives for surgeons to participate in trauma care. PMID- 16307950 TI - Serum bicarbonate may replace the arterial base deficit in the trauma intensive care unit. AB - OBJECTIVE: Arterial base deficit (BD) is a commonly used marker of injury severity and endpoint of resuscitation but requires an arterial puncture and blood gas analysis. Serum bicarbonate (HCO3) is routinely obtained as part of the chemistry panel on most admissions. We hypothesized that serum HCO3 strongly correlates with arterial BD and provides equivalent predictive information. METHODS: All trauma ICU admissions from 1996 to 2004 with simultaneously obtained serum chemistry panels and arterial blood gases were identified. Correlation between BD and HCO3 was analyzed by using linear regression, and predictive abilities for acidoses and mortality were compared using the area under the respective receiver operating characteristic curve (AUC). Separate analyses were done for the entire dataset and the subset of ICU admission laboratory values. RESULTS: We identified 3,102 patients with 50,311 matched pairs of laboratory data. Serum HCO3 showed a significant linear correlation with BD for all laboratory sets (r = 0.85, P < .01) and admission laboratory values only (r = 0.80, P < .01). Serum HCO3 reliably predicted the presence of significant metabolic acidoses (BD >5), with an AUC of 0.96 (P < .01), which clearly outperformed pH (AUC = 0.83), anion gap (AUC = 0.7), and lactate (AUC = 0.73). The mean admission BD among survivors was 2.5 versus 5.2 for nonsurvivors (P < .01), and the mean HCO3 was 17.7 versus 19.8 (P < .01). The admission HCO3 identified nonsurvivors as accurately as BD (AUCs of 0.66 and 0.68) and more accurately than either pH (AUC = 0.53) or anion gap (AUC = 0.6). CONCLUSION: Serum HCO3 measurement shows a strong linear correlation and similar predictive ability compared with the arterial BD. Serum HCO3 may be safely and accurately substituted for arterial BD measurement in critically injured patients. PMID- 16307951 TI - Impact of the 80-hour work week on resident emergency operative experience. AB - BACKGROUND: The goal of this study was to analyze the impact of the 80-hour work week on the emergency operative experience of surgical residents. METHODS: A 2 year retrospective comparison of the operative experience in emergency abdominal procedures of postgraduate year 4 and 5 residents in a city hospital before (group 1) and after (group 2) duty hour restriction. RESULTS: There was no difference between groups in the mean number of procedures performed as the primary surgeon, but group 2 showed a 40% decrease in technically advanced procedures with a 44% increase in basic procedures. The study also demonstrated a 54% decrease in the operative volume as first assistant. Operative continuity of care by residents decreased from 60% to 26% of cases. CONCLUSIONS: The ACGME regulatory environment is adversely affecting the emergency operative experience of surgical residents. Our findings underscore the need to develop alternative methods to augment the residents' operative experience. PMID- 16307952 TI - Can we afford to do laparoscopic appendectomy in an academic hospital? AB - BACKGROUND: Multiple studies have shown laparoscopic appendectomy to be safe for both acute and perforated appendicitis, but there have been conflicting reports as to whether it is superior from a cost perspective. Our academic surgical group, who perform all operative cases with resident physicians, has been challenged to reduce expenses in this era of cost containment. We recognize resident training is an expensive commodity that is poorly reimbursed, and hypothesized laparoscopic appendectomy was too expensive to justify resident teaching of this procedure. The purpose of this study was to determine if laparoscopic appendectomy is more expensive than open appendectomy. METHODS: From April 2003 to April 2004, all patients undergoing appendectomy for presumed acute appendicitis at our university-affiliated teaching hospital were reviewed; demographic data, equipment charge, minutes in the operating room (OR), hospital length of stay, and total hospital charge were analyzed. OR minute charges were gradated based on equipment use and level of skilled nursing care. Conversions to open appendectomy were included in the laparoscopic group for analysis. RESULTS: During the study period, 247 patients underwent appendectomy for preoperative diagnosis of acute appendicitis, with 152 open (113 inflamed, 37 perforated, 2 normal), 88 laparoscopic (69 inflamed, 12 perforated, 7 normal), and 7 converted (2 inflamed, 4 perforated, 1 normal) operations performed. The majority were men (67%) with a mean age of 31.4 +/- 2.2 years. Overall, there was significant difference (P < .05) in intraoperative equipment charge (125.32 dollars +/- 3.99 dollars open versus 1,078.70 dollars +/- 24.06 dollars lap), operative time charge (3,022.16 dollars +/- 57.51 dollars versus 4,065.24 dollars +/- 122.64 dollars), and total hospital charge (12,310 dollars +/- 772 dollars versus 16,773 dollars +/- 1,319 dollars) but no significant difference in operative minutes (56.3 +/- 1.3 versus 57.4 +/- 2.3), operating room minutes (90.5 +/- 1.7 versus 95.7 +/- 2.5), or hospital days (2.6 versus 2.2). In subgroup analysis of patients with uncomplicated appendicitis, open and laparoscopic groups had equivalent hospital days (1.47 versus 1.49) but significantly different hospital charges (9,632.44 dollars versus 14,251.07 dollars). CONCLUSIONS: Although operative time was similar between the 2 groups, operative and total hospital charges were significantly higher in the laparoscopic group. Unless patient factors warrant a laparoscopic approach (questionable diagnosis, obesity), we submit open appendectomy remains the most cost-effective procedure in a teaching environment. PMID- 16307953 TI - Gender differences in general surgical careers: results of a post-residency survey. AB - BACKGROUND: This study's purpose was to gain perspective regarding general surgery career choices while examining gender differences. METHODS: Graduates of a general surgery residency (n = 189) received surveys addressing fellowship training, practice type, case composition, work hours, academic involvement, income, residency, and career satisfaction. RESULTS: Several gender differences were identified. Most men (64%) listed general surgery as their primary work type, whereas women most commonly reported breast surgery (38%, P < .001). More women worked <40 hours per week (25% versus 9%, P = .049). There was no gender difference in income for fellowship-trained surgeons, but a disparity in income >200,000 dollars favored non-fellowship-trained men (74% versus 36%, P = .0031). Both genders reduced work hours. Women reduced them exclusively for personal/family demands. Both genders reported satisfaction with their surgical careers (93%). CONCLUSIONS: Some gender differences in surgery were identified. However, both genders maintain a high level of satisfaction with their career choice. PMID- 16307954 TI - In-house trauma attendings: is there a difference? AB - INTRODUCTION: Outcomes of patients who met trauma activation criteria were examined before and after implementation of in-house attending call. MATERIALS AND METHODS: Outcomes for the out-of-house period (OH) (February 1, 2001 to October 31, 2002) were compared with the in-house period (IH) (November 1, 2002 to June 30, 2004). Measures included overall mortality, length of stay (LOS) in the hospital, intensive care unit (ICU) and emergency department, and preventable deaths. RESULTS: A total of 2,019 trauma activations were studied (1,036 OH, 983 IH). The groups were equivalent on admission. There was no difference in hospital LOS, ICU LOS, ventilator days, or overall mortality. Preventable deaths occurred in 8.1% of the OH group and in 1.0% of the IH group (P < .02). CONCLUSIONS: Aggregate statistics and the use of surrogate markers to determine outcomes may not accurately portray the impact of attending surgeons on the quality of care. Implementation of in-house call resulted in a decreased incidence of preventable deaths. PMID- 16307955 TI - Conceptual accessibility and sentence production in a free word order language (Odawa). AB - The study reported here was conducted in the Algonquian language of Odawa (a.k.a. Ottawa), with the goal of gaining new insight into the ways that conceptual accessibility affects human sentence production. The linguistic characteristics of Odawa are quite different from those found in the languages most often examined by psycholinguists. The data obtained from the sentence production experiment reported here are thus relevant to production in a heretofore unexamined language. Moreover, the data inform broader theoretical issues, such as the extent to which sentence production can be considered as an incremental process, and the interaction of the various factors affecting conceptual accessibility. In addition, the study stands as evidence that experimental psycholinguistic research can and should be carried out in typologically diverse languages. PMID- 16307956 TI - Infants' understanding of object-directed action. AB - When and in what ways do infants recognize humans as intentional actors? An important aspect of this larger question concerns when infants recognize specific human actions (e.g. a reach) as object-directed (i.e. as acting toward goal objects). In two studies using a visual habituation technique, 12-month-old infants were tested to assess their recognition that an adult's reach is directed toward its target object. Infants in the experimental condition were habituated to a display in which an actor reached over a wall-like barrier with an arcing arm movement, to pick up a ball. After habituation infants saw two test displays, for which the barrier was removed. In the direct test event the actor reached directly for the ball, the arm tracing a visually new path, but the action consistent with attempting to reach for the object as directly as possible. In the indirect test event the actor traced the old path, reaching over in an arc, even though the wall was no longer present. This arm movement was identical to that in habituation but no longer displayed a reach going directly to its object. In a control condition infants saw the same movements but in a situation with no goal-object. In the experimental conditions, with a goal object present, infants looked longer at the indirect test event in comparison to the direct test event. In the control conditions infants looked equally at both indirect and direct test events. We conclude that sensitivity to human object-directed action is established by 12-month-olds and compare these results to recent findings by [Gergely, G., Nadasdy, Z., Csibra, G., & Biro S. (1995). Taking the intentional stance at 12 months of age. Cognition, 56, 165-193] and [Woodward, A. (1998). Infants selectively encode the goal object of an actor's reach. Cognition, 69, 1 34]. PMID- 16307957 TI - Applying global workspace theory to the frame problem. AB - The subject of this article is the frame problem, as conceived by certain cognitive scientists and philosophers of mind, notably Fodor for whom it stands as a fundamental obstacle to progress in cognitive science. The challenge is to explain the capacity of so-called informationally unencapsulated cognitive processes to deal effectively with information from potentially any cognitive domain without the burden of having to explicitly sift the relevant from the irrelevant. The paper advocates a global workspace architecture, with its ability to manage massively parallel resources in the context of a serial thread of computation, as an answer to this challenge. Analogical reasoning is given particular attention, since it exemplifies informational unencapsulation in its most extreme form. Because global workspace theory also purports to account for the distinction between conscious and unconscious information processing, the paper advances the tentative conclusion that consciousness may go hand-in-hand with a solution to the frame problem in the biological brain. PMID- 16307958 TI - Capacity limits for face processing. AB - We present three experiments in which subjects were asked to make speeded sex judgements (Experiment 1) or semantic judgements (Experiments 2 and 3) to face targets and nonface items, while ignoring a solitary flanking distractor face or a nonface stimulus. Distractors could be either congruent (same response category) or incongruent (different response category) with the target. Distractor congruency effects were consistently observed in all combinations of target-distractor stimulus pairs, except when a distractor face flanked a target face. The failure to find congruency effects in this condition was explored further in a fourth experiment, in which four task-irrelevant flankers were simultaneously presented. Once again, no face-face congruency effects were found, even though comparison distractors interfered with face and nonface targets alike. However, four simultaneously presented distractor faces did not interfere with nonface targets either. We suggest that these experiments demonstrate a capacity limit for visual processing in these conditions, such that no more than one face is processed at a time. PMID- 16307959 TI - Is family planning a subspecialty of obstetrics and gynecology? PMID- 16307960 TI - Revisiting community-based distribution programs: are they still needed? AB - Community-based distribution (CBD) programs are the optimum way of reaching people in rural areas of developing countries where conventional methods of delivery do not exist or fail. This paper reviews findings and experiences from over 30 years of efforts to implement CBD of family planning methods around the world. Although research suggests that community-based service delivery can contribute to contraceptive use, the magnitude of impact is often in doubt or its existence is questionable when compared to alternative family planning delivery services. After the review of more than 30 years of CBD work, we found that these programs are still needed to meet the needs for contraception in rural communities and isolated city neighborhoods in developing countries. Integration with other health outreach programs, effective management, keeping training of agents brief and letting them distribute contraceptives and keeping all or part of the profits instead of paying them salaries are some of the strategies that can make CBD programs more efficient. PMID- 16307961 TI - Will microbicide trials yield unbiased estimates of microbicide efficacy? AB - OBJECTIVE: To evaluate the effect of adherence and condom use on apparent efficacy of microbicides. DESIGN: Hypothetical trial designs and scenarios. METHODS: Mathematical calculations of effectiveness. RESULTS: In a randomized trial of a candidate microbicide and a placebo, nonuse of the microbicide will result in underestimation of microbicide efficacy, with the magnitude of this difference between effectiveness and efficacy increasing directly with the level of microbicide nonuse. Adding condoms to the trial will not change this expected result as long as use of condoms and microbicide is independent, and microbicide use is the same in the trials with and without condoms. However, if microbicide use is lower in the trial with condoms, then effectiveness will be lower than in the trial without condoms, with the magnitude of the difference between effectiveness and efficacy being even greater. Moreover, condom and microbicide use may not be independent. If participants tend to use condoms rather than nothing, the trial result will more closely approximate microbicide efficacy. If, however, participants substitute condom use for microbicide use, then the expected estimate of effectiveness will less closely approximate microbicide efficacy and could be closer to or further away from the expected estimate of effectiveness than in the trial without condoms. In another trial design, where there is either simultaneous use of microbicide and condoms or no use of either (guaranteed when condoms are packaged with either a microbicidal or a placebo gel), expected effectiveness will fall short of microbicide efficacy. If nonuse is the same in a trial without condoms and a trial with microbicidally lubricated condoms, then the trial with condoms will produce an estimate of effectiveness that less closely approximates microbicide efficacy than would the trial without condoms. If there is less nonuse in the trial with condoms, then the expected estimate of effectiveness will more closely approximate microbicide efficacy and could be closer to or further away from the expected estimate of effectiveness than in the trial without condoms. CONCLUSIONS: Nonuse of a microbicide and use of condoms may seriously impair our ability to identify an effective microbicide. PMID- 16307962 TI - Treatment of premenstrual dysphoric disorder with a new drospirenone-containing oral contraceptive formulation. AB - PURPOSE: This multicenter, double-blind, placebo-controlled crossover study evaluated the efficacy of a new oral contraceptive (OC) formulation containing drospirenone 3 mg and ethinyl estradiol (EE) 20 mug in treating symptoms of premenstrual dysphoric disorder (PMDD). METHOD: The OC formulation or placebo was administered for 24 days in a 28-day cycle (24/4), rather than the usual 21-day active treatment, 7-day inert-pill regimen. Participants (N=64) were randomized to either study treatment for three cycles and then after a washout period of one treatment-free cycle switched to the alternate treatment. RESULTS: The mean decrease from baseline for total Daily Record of Severity of Problems (DRSP) scores while using drospirenone/EE was significantly greater than for placebo ( 12.47, 95% CI=-18.28, -6.66; p<.001). A positive response (i.e., a score of 1 or 2 in the Clinical Global Impressions-Improvement scale) occurred in 61.7% and 31.8% of subjects while taking drospirenone/EE and placebo, respectively (p=.009). CONCLUSION: Drospirenone/EE, given in a 24/4 regimen, was superior to placebo for improving symptoms associated with PMDD. PMID- 16307963 TI - Postpartum contraception: the New Mexico Pregnancy Risk Assessment Monitoring System. AB - OBJECTIVE: To examine factors associated with postpartum contraception, including the relationship between ethnicity and postpartum contraceptive use. METHODS: We used data from the New Mexico Pregnancy Risk Assessment Monitoring System, which monitors selected maternal events occurring before, during and after pregnancy. RESULTS: Our findings in 4096 women revealed that women who are aged >or=35 years, unmarried and lacking a postpartum visit have increased risk of no postpartum contraception. The odds of postpartum contraception were over three times greater in women with a postpartum visit [adjusted odds ratio (OR)=3.06, 95% confidence interval (CI): 2.17-4.31) and over 50% greater in married women (adjusted OR=1.57, 95% CI: 1.16-2.11). Hispanic women were more likely than were Native Americans to use postpartum contraception (OR=1.25, 95% CI: 0.95-1.64). CONCLUSION: Focused contraception counseling, especially in the postpartum setting, is important to help ensure the well-being of women and children. PMID- 16307964 TI - Barriers to intrauterine device insertion in postpartum women. AB - OBJECTIVE: The objective of this study was to determine the proportion of postpartum women at the University of New Mexico who choose an IUD for contraception, the number who actually obtain one and the barriers to postpartum IUD insertion. METHOD: We conducted a retrospective chart review of 1627 postpartum women who delivered at the University of New Mexico. Those women who indicated at hospital discharge that they desired an IUD comprised the study group of 193 women. Medical records were reviewed to identify the timing of IUD placement. If an IUD was not inserted, we attempted to determine the reason by reviewing clinic records. RESULTS: Twelve percent of postpartum women requested an IUD. Records were available for 114 women. Of these, only 69 (60%) actually obtained an IUD. Barriers to postpartum IUD insertion included provider advice against the IUD, patient failure to return for a postpartum visit and early repeat pregnancy. CONCLUSION: We conclude that postpartum women desiring an IUD may have difficulty obtaining one. PMID- 16307965 TI - Spontaneous upward movement of lowly placed T-shaped IUDs. AB - OBJECTIVE: To assess the possible progression of intrauterine devices (IUDs) to a lower or upper position in women. MATERIAL AND METHODS: Thirty-two lowly placed T shaped IUDs in women with 0-3 children were examined with transvaginal ultrasonography after 2 months of insertion in order to assess the possible progression of the IUDs. Also, the IUDs inserted in women with 0-3, 0-1 and 2-3 children were compared to determine whether this progression was different depending on parity. RESULTS: Intrauterine devices (97%, 100% and 93.7%) in the group of women with 0-3, 0-1 and 2-3 children, respectively, moved upward. The mean decrease in IUD-fundus distance was 6.8, 7.4 and 4.9 mm, respectively. CONCLUSION: Lowly inserted T-shaped IUDs tend to move upward after insertion. Therefore, initial concern about low placement of a T-shaped IUD is not justified as most of them spontaneously readjust their position. PMID- 16307966 TI - Tamoxifen treatment of bleeding irregularities associated with Norplant use. AB - OBJECTIVES: To evaluate the possible role of tamoxifen (selective estrogen receptor modulators, SERM) in treating bleeding irregularities associated with Norplant contraceptive use. MATERIAL AND METHODS: Randomized clinical trial including 100 Norplant users complaining of vaginal bleeding irregularities. The trial was conducted in the Family Planning Clinic of Assiut University Hospital. Women were assigned at random to receive tamoxifen tablets (10 mg) twice daily for 10 days or similar placebo. Women were followed-up for 3 months. The end points were percentage of women who stopped bleeding during treatment, bleeding/spotting days during the period of follow-up, effect of treatment on their lifestyle, and side effects and discontinuation of contraception. RESULTS: There was good compliance with treatment. At the end of treatment, a significantly higher percentage of tamoxifen users stopped bleeding in comparison to the control group (88% vs. 68%, respectively; p=.016). Women who used tamoxifen had significantly less bleeding and/or spotting days than women who used placebo, during the first and second months. During the third month, there were no significant differences between the two groups. Women who used tamoxifen reported improvement in performing household activities, religious duties and in sexual life, during the first 2 months. In the third month, there were no differences between the two groups. There were no significant differences between tamoxifen and placebo groups in reporting side effects. In the group who used tamoxifen, two women discontinued Norplant use because of bleeding vs. nine women in the placebo group. CONCLUSION: Tamoxifen use at a dose of 10 mg twice daily orally, for 10 days, has a beneficial effect on vaginal bleeding associated with Norplant use. In addition, the bleeding pattern was better in women who used tamoxifen for the following 2 months after treatment. However, these results have to be confirmed in a larger trial before advocating this line of treatment. PMID- 16307967 TI - A randomized controlled study comparing 600 versus 1,200 microg oral misoprostol for medical management of incomplete abortion. AB - OBJECTIVES: Although a number of studies have shown misoprostol's promise as a nonsurgical treatment for incomplete abortion, few have systematically examined treatment protocols. This study documents the effectiveness of 600 versus 1,200 microg oral misoprostol for this indication. METHODS: From May 2002 to January 2003, 300 women with incomplete abortion were recruited at a large tertiary facility in Vietnam and randomized to either a single-dose (600 microg) or a repeated-dose (600 microg x 2) regimen of oral misoprostol for the treatment of their condition. RESULTS: Misoprostol effectively evacuated the uterus for nearly all women (94.6%; n=279), with most reporting bleeding for 4 days (+/-2.3) and pain/cramps lasting 1 day (+/-1.0). Women indicated that the side effects were tolerable (96%) and that their experience was satisfactory (95%). CONCLUSIONS: Oral misoprostol (600 or 1,200 microg) offers a safe, effective and acceptable treatment for incomplete abortion. Larger studies to assess the advantages and disadvantages of misoprostol as compared with standard surgical care are needed to assess its role in postabortion care programs worldwide. PMID- 16307968 TI - Effect on insulin sensitivity of Implanon vs. GnRH agonist in women with endometriosis. AB - OBJECTIVES: To evaluate the influence of two medical treatments for endometriosis on insulin sensitivity. STUDY DESIGN: After surgery, 26 women with endometriosis were randomly allocated to a 6-month treatment with a GnRH agonist (Leuprorelin 3.75 mg/28 days) or a subdermal progestin implant (etonogestrel 68 mg). Insulin sensitivity (SI) and glucose utilization independent of insulin (Sg) were investigated at baseline and after 6 months by a frequently sampled intravenous glucose tolerance test (FSIGT) associated with the minimal model method. RESULTS: Both therapies tended to decrease SI, but the effect did not reach statistical significance in the GnRH agonist group (5.43+/-1.29 vs. 3.99+/-0.8) and was significant in the etonogestrel group (5.74+/-1.12 vs. 3.95+/-0,78; p=.046). Sg, fasting glucose, insulin, C-peptide and C-peptide/insulin were not modified by either treatment. CONCLUSIONS: The modifications of glucose-insulin metabolism induced by the GnRH agonist are of no relevance for the short-term use of this molecule. Even if the modification induced by the etonogestrel implant is subtle and of no major impact, it should be taken into consideration for the long-term treatment of individuals with abnormalities of glucose-insulin metabolism. PMID- 16307969 TI - Spermicidal activity of dermaseptins. AB - OBJECTIVE: The present study was undertaken to elucidate the spermicidal efficacy of two synthetic antimicrobial peptides, dermaseptin (DS1 and DS4). METHODS: Twenty samples of fresh semen were obtained from patients aged between 23 and 35 years. The ability of DS to kill sperm was evaluated by the Sander-Cramer test under in vitro conditions. RESULTS: The data showed that sperm motility was inhibited with various concentrations of DS at different intervals ranging from 2 to 240 min. The effective 100% inhibitory concentration (EC(100)) of DS4 in 2 min during the sperm immobilization assay was 100 microg/mL whereas the sperm immobilization of EC(100) of DS1 was 200 microg/mL. The presence of 0.1% chelating agent ethylenediaminetetraacetic acid (EDTA) reduced the EC(100) of DS4 to 10 microg/mL whereas less than a two-time enhancement in DS1 activity was observed upon combination with EDTA. The action of DSs on sperm motility was observed to be dose dependent. Supplementation with pentoxifylline and that with calcium are known to enhance the motility of sperm but they did not prevent the spermicidal action of DSs. CONCLUSION: This present study indicates that DS is an effective agent to kill sperm. In view of this fact, it is suggested that DS4 has antibacterial, antiviral, antifungal and potentially spermicidal activities and could be a potent vaginal contraceptive. PMID- 16307970 TI - Evaluation of contraceptive activity of a mineralo-herbal preparation in Sprague Dawley rats. AB - OBJECTIVE: This study was aimed to evaluate a marketed mineralo-herbal preparation containing plants known to have potent contraceptive activity, or contraindicated for use during pregnancy in folklore/ancient Indian literature and recommended for use as an appetizer and headache, hyperacidity and chronic constipation reliever for effect on spermatogenesis and implantation-cum-early postimplantation events in adult Sprague-Dawley rats. METHODS: The preparation, suspended in distilled water with the addition of sterile gum acacia, was administered at 1 g/kg daily dose (extrapolated from human dose on surface area basis) to male rats covering one spermatogenic cycle and to female rats during the entire preimplantation and early postimplantation period by oral route. Fertility performance of male rats was tested following mating with untreated fertile females. RESULTS: Findings of this study indicate that the mineralo herbal preparation at this dose and schedule produced no discernible effect on weight of testis, epididymis and accessory glands, spermatogenesis, vasal sperm picture or mating rate in male rats when administered during the period covering one spermatogenic cycle, but caused significant reduction in number of implantations in females mated with these male rats as well as in female rats treated during the postcoital period. CONCLUSIONS: Any adverse effect on fertility/reproductive health following administration over longer periods/at higher doses in humans habituated to continuous use of this preparation cannot be completely ruled out from this limited study. Findings also suggest caution in indiscriminate use of this and other such preparations containing varying amounts of plants/plant products reported to possess contraceptive property and available for other pharmacological indications over-the-counter in most countries. PMID- 16307971 TI - Contraceptive responses of female hamsters immunized with recombinant sperm protein P26h. AB - BACKGROUND: A number of antigens have been characterized and proposed as potential candidates for immunocontraception. P26h, a 26-kDa hamster sperm protein located on the acrosomal cap, is known to be involved in sperm-zona pellucida interactions. Furthermore, in vivo fertilization can be blocked by active immunization of male hamsters against P26h or maltose-binding protein recombinant P26h (MBP-P26h). OBJECTIVE: The aim of this study was to investigate the immune response and reproductive function of female hamsters immunized against MBP-P26h. RESULTS: Active immunization against MBP-P26h resulted in anti P26h circulating antibodies, with enzyme-linked immunosorbent assay (ELISA) titers showing interindividual variability. The antibodies produced by the animals immunized against MBP-P26h reacted with the native P26h protein in ELISA, in Western blot analysis and in immunostaining performed on cauda epididymal spermatozoa. Mating of immunized female hamsters resulted in a significant decrease in the number of viable fetuses only in females with high titers of anti P26h circulating antibodies. DISCUSSION: This result is in agreement with the sperm-zona pellucida binding assay's results. Indeed, sera collected from immunized animals, and not from control animals, significantly blocked sperm-zona pellucida binding in vitro. Histological studies showed that active immunization did not cause any pathology in the reproductive tissues. CONCLUSION: These findings suggest that P26h is a potential candidate for the development of a contraceptive vaccine in both males and females. PMID- 16307972 TI - Tetrachlorodibenzo-p-dioxin in baby food made from chicken produced before and after the termination of ball clay use in chicken feed in the United States. AB - Polychlorodibenzo-p-dioxin/furans were determined in chicken-containing baby foods collected from an annually conducted total diet survey by the US FDA during the last half of fiscal year (FY) 1997 through the first half of FY 1998. 2,3,7,8 Tetrachlorodibenzo-p-dioxin (TCDD) was found in 8 of 11 baby food samples. The levels were between 0.025 and 0.28 ng kg(-1) wet wt (0.25-5 ng kg(-1) lipid). The mean TCDD value for chicken-containing baby food with "nondetects" equal to 0 was 1.2 ng kg(-1) lipid, eight times higher than the average level found during a previous survey of chicken lipid TCDD levels in 1996. All other 2,3,7,8-chlorine substituted dibenzo-p-dioxin congeners were also found with a profile consistent with the use of ball clay in chicken feed. TCDD was not detected in any FY 2000 baby foods made with chicken, with an average limit of detection (LOD) of 0.025 ng kg(-1) wet wt. Whole eggs collected in 1997 from producers that never used ball clay in feed revealed TCDD measurements that were nearly all nondetects and none above the median LOD of 0.015 ng kg(-1) wet wt. The percentage of chickens fed ball clay in their feed was estimated to be between 2.6% and 3.5% of all chickens produced in the United States in 1997. PMID- 16307973 TI - Concentrations of selective metabolites of organophosphorus pesticides in the United States population. AB - We report population-based concentrations (stratified by age, sex, and composite race/ethnicity variables) of selective metabolites of chlorpyrifos (3,5,6 trichloro-2-pyridinol; TCPY), chlorpyrifos methyl (TCPY), malathion (malathion dicarboxylic acid; MDA), diazinon (2-isopropyl-4-methyl-6-hydroxypyrimidine; IMPY), methyl parathion (para-nitrophenol; PNP), and parathion (PNP). We measured the concentrations of TCPY, MDA, IMPY, and PNP in 1997 urine samples from participants, aged 6-59 years, of the National Health and Nutrition Examination Survey, 1999-2000. We detected TCPY in more than 96% of the samples tested. Other organophosphorus pesticide metabolites were detected less frequently: MDA, 52%; IMPY, 29%; and PNP, 22%. The geometric means for TCPY were 1.77 microg/L and 1.58 microg/g creatinine. The 95th percentiles for TCPY were 9.9 microg/L and 8.42 microg/g creatinine. The 95th percentiles for MDA were 1.6 microg/L and 1.8 microg/g creatinine. The 95th percentiles for IMPY and PNP were 3.7 microg/L (3.4 microg/g creatinine) and 5.0 microg/L (4.2 microg/g creatinine), respectively. Multivariate analyses showed that children aged 6-11 years had significantly higher concentrations of TCPY than adults and adolescents. Similarly, adolescents had significantly higher TCPY concentrations than adults. Although the concentrations between sexes and among composite racial/ethnic groups varied, no significant differences were observed. PMID- 16307974 TI - Effects of methyltestosterone on reproduction in the Japanese quail (Coturnix coturnix japonica). AB - Although the hormone-mediated effects of the synthetic androgenic hormone methyltestosterone (MT) are well characterized in mammals, little is known about endocrine and other toxic effects on reproduction in birds. In a one-generation study, MT was administered to adult Japanese quail (12 pairs per group) at dietary dose levels of 0, 10, 50, and 110 ppm for a period of 3 weeks. Reproductive performance was severely affected in the groups receiving 50 and 110 ppm MT. In females, the egg-laying rate was reduced not only related to the dose administered but also to the duration of treatment. The administration of 110 ppm, and to a lesser extent, of 50 ppm MT resulted in an immediate and dramatic decrease in the total number of eggs laid, which complicated reliable assessment of other reproduction-related parameters. In males, the findings suggested inhibition of spermatogenesis at dose levels of 50 ppm and above, resulting in a subsequent reduction in male fertility. PMID- 16307975 TI - Enhanced oxidative stress and endothelial dysfunction in streptozotocin-diabetic rats exposed to fine particles. AB - The association between ambient particulate matter (PM) and cardiovascular diseases has been demonstrated in epidemiological studies. Recent studies suggest that diabetic patients are at greater risk for PM-associated cardiovascular events. Although diabetes and PM exposure individually have been reported to be associated with increased oxidative stress, inflammation, and endothelial dysfunction, it is not clear whether PM may induce synergistic interaction effects on these parameters in diabetics. Strepotozotocin-induced diabetic (n=4) and healthy (n=4) rats were intratracheally administered with PM2.5 collected from a busy traffic area in a dose of 200 microg suspended in 0.5 mL phosphate buffered saline (PBS). The same number of rats was exposed to PBS as controls. Cell and differential counts and protein and lactate dehydrogenase activity were determined in bronchoalveolar lavage. Markers of 8-hydroxydeoxy-guanosine (8 OHdG), endothelin-1 (ET-1), and [nitrate+nitrite], an indicator of nitric oxide (NO) production, in addition to C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) in peripheral blood were also determined. Our results showed that diabetic rats were associated with increased 8-OHdG, IL-6, and ET-1 decreased [nitrate+nitrite]. In nondiabetic rats PM exposure was also associated with increased 8-OHdG, IL-6, TNF-alpha, and CRP but decreased [nitrate+nitrite]. Interestingly, increases of 8-OHdG and ET-1 after PM exposure were more prominent in diabetic rats than in nondiabetic rats. The general linear model further indicated that there were interactions between diabetes and PM on 8-OHdG (P<0.01) and ET-1 (P=0.08). We suggest that PM exposure may enhance the risk of cardiovascular diseases through interaction between PM and diabetes on excess reactive oxygen species generation and endothelial dysfunction. These findings provide further support for previous epidemiological studies. PMID- 16307976 TI - Winter urban air particles from Rome (Italy): effects on the monocytic macrophagic RAW 264.7 cell line. AB - Epidemiological data show an association between exposure to elevated levels of particulate matter (PM), in particular the fine fraction (<2.5 microm in diameter), and an increase in cardiovascular mortality and respiratory symptoms. The aim of this study was to compare the in vitro toxicity of coarse and fine particulate matter collected with a cascade impactor during winter in an urban area of Rome in relation to their physicochemical characterization (size distribution and chemical composition) as assessed by analytical electron microscopy (SEM/EDX). The X-ray microanalysis data of single particles of coarse and fine matter were analyzed by hierarchical cluster analysis to determine the principal component of the two granulometric fractions. The main chemical difference between the two fractions was the greater abundance of carbonaceous particles in the fine fraction. We compared the ability of coarse and fine fractions, carbon black (CB), and residual oil fly ash (ROFA) to induce arachidonic acid release and tumor necrosis factor-alpha (TNF-alpha) production in the monocytic-macrophagic RAW 264.7 cell line at concentrations of 30 and 120 microg/mL. Our results showed that CB and ROFA were consistently less effective than both fractions of urban particles at inducing an inflammatory reaction in RAW 264.7 cells. Both PM fractions dose-dependently increased TNF-alpha production in RAW 264.7 cells after 5 and 24h of incubation, and only the TNF alpha production induced by coarse particles at 30 microg/mL decreased significantly (P<0.01) after 24h of treatment. In our in vitro model the winter fine fraction was more reactive than the winter coarse fraction, in contrast to a previously examined summer sample. In the summer sample, coarse particles produced higher levels of inflammatory mediators than fine particles and the CB was consistently less effective than the urban particles. The different behaviors between summer and winter urban fractions may be due to their different physicochemical characteristics; in fact, the comparison of the two samples' characterization by SEM/EDX and X-ray photoelectron spectroscopy (XPS) analysis showed that in winter the carbonaceous particles are more abundant than in summer and that winter particles carry a greater quantity of organic compounds. We suggest that the higher concentration of organic compounds on fine carbonaceous particles may partially explain the higher activation of RAW 264.7 cells by fine particles. PMID- 16307977 TI - N-acetylcysteine and deferoxamine reduce pulmonary oxidative stress and inflammation in rats after coal dust exposure. AB - Coal dust inhalation induces oxidative damage and inflammatory infiltration on lung parenchyma. Thus, the aim of this study was to determine whether N acetylcysteine (NAC) administered alone or in combination with deferoxamine (DFX), significantly reduced the inflammatory infiltration and oxidative damage in the lungs of rats exposed to coal dust. Forty-two male Wistar rats (200-250 g) were exposed to the coal dust (3mg/0.5 mL saline, 3 days/week, for 3 weeks) by intratracheal instillation. The animals were randomly divided into three groups: saline 0.9% (n=8), supplemented with NAC (20mg/kg of body weight/day, intraperitoneal injection (i.p.)) (n=8), and supplemented with NAC (20 mg/kg of body weight/day, i.p.) plus DFX (20 mg/kg of body weight/week) (n=8). Control animals received only saline solution (0.5 mL). Lactate dehydrogenase activity and total cell number were determined in the bronchoalveolar lavage fluid. We determined lipid peroxidation and oxidative protein damage parameters and catalase and superoxide dismutase activities in the lungs of animals. Intratracheal instillation of coal dust in the lungs of rats led to an inflammatory response and induced significant oxidative damage. The administration of NAC alone or in association with DFX reduced the inflammatory response and the oxidative stress parameters in rats exposed to coal dust. PMID- 16307978 TI - Asian sand dust enhances ovalbumin-induced eosinophil recruitment in the alveoli and airway of mice. AB - Asian sand dust (ASD) containing sulfate (SO4(2-)) reportedly causes adverse respiratory health effects but there is no experimental study showing the effect of ASD toward allergic respiratory diseases. The effects of ASD and ASD plus SO4(2-) toward allergic lung inflammation induced by ovalbumin (OVA) were investigated in this study. ICR mice were administered intratracheally with saline; ASD alone (sample from Shapotou desert); and ASD plus SO4(2-) (ASD-SO4); OVA+ASD; OVA+ASD-SO4. ASD or ASD-SO4 alone caused mild nutrophilic inflammation in the bronchi and alveoli. ASD and ASD-SO4 increased pro-inflammatory mediators, such as Keratinocyte chemoattractant (KC) and macrophage inflammatory protein (MIP)-1 alpha, in bronchoalveolar lavage fluids (BALF). ASD and ASD-SO4 enhanced eosinophil recruitment induced by OVA in the alveoli and in the submucosa of the airway, which has a goblet cell proliferation in the bronchial epithelium. However, a further increase of eosinophils by addition of SO4(2-) was not observed. The two sand dusts synergistically increased interleukin-5 (IL-5) and monocyte chemotactic protein-1 (MCP-1), which were associated with OVA, in BALF. However, the increased levels of IL-5 were lower in the OVA+ASD-SO4 group than in the OVA+ASD group. ASD caused the adjuvant effects to specific-IgG1 production by OVA, but not to specific-IgE. These results suggest that the enhancement of eosinophil recruitment in the lung is mediated by synergistically increased IL-5 and MCP-1. IgG1 antibodies may play an important role in the enhancement of allergic reaction caused by OVA and sand dust. However, extra sulfate may not contribute to an increase of eosinophils. PMID- 16307979 TI - The effect of extremely low-frequency electromagnetic fields on skin and thyroid amine- and peptide-containing cells in rats: an immunohistochemical and morphometrical study. AB - The aim of this study was to investigate the influence of extremely low-frequency electromagnetic fields (ELF-EMFs) on mast cells (MCs), parafollicular cells, and nerve fibers in rat skin and thyroid gland. The experiment was performed on 24 2 month-old Wistar male rats exposed for 4h a day, 7 days a week for 1 month to EMFs (50 Hz, 100-300 microT, 54-160 V/m). After sacrifice, samples of skin and thyroid were processed for indirect immunohistochemistry or toluidine blue staining and then were analyzed using the methods of stereology. The antibody markers to serotonin, substance P, calcitonin gene-related peptide (CGRP), and protein gene product 9.5 (PGP) were applied to skin sections and PGP, CGRP, and neuropeptide Y (NPY) markers to the thyroid. A significantly increased number of serotonin-positive MCs in the skin and NPY-containing nerve fibers in the thyroid of rats exposed to ELF-EMF was found compared to controls, indicating a possible EMF effect on skin and thyroid vasculature. PMID- 16307980 TI - Alterations in carbohydrates and the protein metabolism of the harmful freshwater vector snail Lymnaea acuminata induced by the Euphorbia tirucalli latex extract. AB - To know the short- as well as long-term effect of aqueous latex extracts of Euphorbia tirucalli on carbohydrate and protein metabolism, the snail Lymnaea acuminata was exposed to sublethal doses of 0.37 and 0.55 mg/L for a 24-h and 0.20 and 0.31 mg/L for a 96-h exposure period. Significant (P<0.05) alterations in the glycogen, pyruvate, lactate, total protein, and free amino acid level, as well as in the activity of enzyme lactic dehydrogenase, succinic dehydrogenase, cytochrome oxidase, protease, aspartate aminotransaminase, and alanine aminotransaminase were observed in the nervous, hepatopancreatic, and ovotestis tissues of the freshwater vector snail L. acuminata exposed to sublethal doses of E. tirucalli latex extract. The alterations in all biochemical parameters were significantly (P<0.05) time and dose dependent. After the 7th day of the withdrawal of treatment, there was significant (P<0.05) recovery in glycogen, pyruvate, lactate, total protein, and the free amino acid level and in the activity of the lactic dehydrogenase, succinic dehydrogenase, cytochrome oxidase, protease, aspartate aminotransaminase and alanine aminotransaminase enzymes in all three of the studied tissues of the snail, which supports the view that the plant product is safe for use as a molluscicide for the control of harmful freshwater vector snails in the aquatic environment. PMID- 16307981 TI - Lysosomal responses as a diagnostic tool for the detection of chronic petroleum pollution at Todos os Santos Bay, Brazil. AB - Coastal marine environments, especially semienclosed systems such as bays, are under unrelenting stress caused by urban and industrial development. Biomonitoring plays a vital role in strategies to identify, assess, and control stressors. However, due to the magnitude of the challenge there is a demand for new and innovative approaches to provide timely and accessible information to environmental managers and policy makers. The present work aimed to assess hydrocarbon levels in sediments from petroleum-related industrial areas at Todos os Santos Bay (Brazil) and associate them to toxicity-induced responses (neutral red retention (NRR) assay) by the burrowing clam Anomalocardia brasiliana. Surface sediments collected during the dry and rainy seasons were analyzed for aliphatic and aromatic hydrocarbons. At the control site, hydrocarbon levels were low and mainly biogenic. The aliphatic hydrocarbon ("total unresolved complex mixture," alkanes, and isoprenoids) concentrations indicated a chronic situation with very little "fresh" oil contamination at the oil-related sites. The polycyclic aromatic hydrocarbons indicated sites moderately contaminated by chronic oil and some pyrolytic input. The effects of those contaminants were assessed by the lysosomal NRR assay applied to A. brasiliana hemocytes. Sediment toxicity at the oil-related sites was evidenced by the lowered capacity of the lysosomes to retain the neutral red dye compared to results from the control site. This research indicates that the NRR assay is a useful and efficient screening technique able to discriminate polluted from clean sites. PMID- 16307982 TI - DDT serum concentration and menstruation among young Chinese women. AB - High DDE and DDT concentrations were found to be associated with shortened menstrual cycle length in Laotian immigrants to the United States. We examined this issue in a sample of young Chinese women. A total of 60 women aged 20-24 years were enrolled in three maternal and child health clinics (20 from urban, 20 from suburban, 20 from rural) in Shanghai, China, and vicinity, in 1998. Of these women, 47 who did not use hormonal contraceptives and had valid menstrual cycle characteristics were included in the analysis for associations among serum DDE and DDT concentration and menstrual cycle length, duration of menses, and heaviness of menstrual flow. In univariate analysis, higher p,p'-DDE concentration was associated with longer menstrual cycle length (0.66 day per 10 microg/L, 95% confidence interval [CI]: 0.21, 1.11 day). With adjustment for age, body mass index, education, occupation, and resident location, the estimate was 0.42 day (95% CI: -0.35, 1.19 day). p,p'-DDE was not associated with duration of menses or heaviness of menstrual flow. Neither p,p'-DDT nor o,p'-DDT were associated with menstrual cycle length, duration of menses or heaviness of menstrual flow. The study largely suggests no association between DDE and DDT concentrations and menstrual cycle characteristics in young Chinese women, though the weak-to-no correlation of DDE with menstrual cycle length merits further study. PMID- 16307983 TI - Heavy metals in commercial fish in New Jersey. AB - Levels of contaminants in fish are of particular interest because of the potential risk to humans who consume them. While attention has focused on self caught fish, most of the fish eaten by the American public comes from commercial sources. We sampled 11 types of fish and shellfish obtained from supermarkets and specialty fish markets in New Jersey and analyzed them for arsenic, cadmium, chromium, lead, manganese, mercury, and selenium. We test the null hypothesis that metal levels do not vary among fish types, and we consider whether the levels of any metals could harm the fish themselves or their predators or pose a health risk for human consumers. There were significant interspecific differences for all metals, and no fish types had the highest levels of more than two metals. There were few significant correlations (Kendall tau) among metals for the three most numerous fish (yellowfin tuna, bluefish, and flounder), the correlations were generally low (below 0.40), and many correlations were negative. Only manganese and lead positively were correlated for tuna, bluefish, and flounder. The levels of most metals were below those known to cause adverse effects in the fish themselves. However, the levels of arsenic, lead, mercury, and selenium in some fish were in the range known to cause some sublethal effects in sensitive predatory birds and mammals and in some fish exceeded health-based standards. The greatest risk from different metals resided in different fish; the species of fish with the highest levels of a given metal sometimes exceeded the human health guidance or standards for that metal. Thus, the risk information given to the public (mainly about mercury) does not present a complete picture. The potential of harm from other metals suggests that people not only should eat smaller quantities of fish known to accumulate mercury but also should eat a diversity of fish to avoid consuming unhealthy quantities of other heavy metals. However, consumers should bear in mind that standards have a margin of safety. PMID- 16307984 TI - Acute health problems among subjects involved in the cleanup operation following the Prestige oil spill in Asturias and Cantabria (Spain). AB - The purpose of this study was to evaluate exposure conditions and acute health effects in subjects participating in the Prestige oil spill cleanup activities and the association between these and the nature of the work and use of protection devices in the regions of Asturias and Cantabria (Spain). The sample comprised 400 subjects in each region, selected from a random sampling of all persons involved in cleanup activities, stratified by type of worker and number of working days. Data were obtained via a structured questionnaire and included information on specific tasks, number of working days, use of protective materials, and acute health effects. These effects were classified into two broad groups: injuries and toxic effects. Data analysis was performed using complex survey methods. Significant differences between groups were evaluated using Pearson's chi(2) test. Unconditional logistic regression was used to compute odds ratios and 95% confidence intervals. Bird cleaners accounted for the highest prevalence of injuries (19% presented with lesions). Working more than 20 days in highly polluted areas was associated with increased risk of injury in all workers. Occurrence of toxic effects was higher among seamen, possibly due to higher exposure to fuel oil and its components. Toxic effects were more frequent among those working longer than 20 days in highly polluted areas, performing three or more different cleaning activities, having skin contact with fuel oil on head/neck or upper limbs, and eating while in contact with fuel or perceiving disturbing odors. No severe disorders were identified among individuals who performed these tasks. However, potential health impact should be considered when organizing cleanup activities in similar environmental disasters. PMID- 16307985 TI - Pericardiodiaphragmatic rupture and cardiac herniation after multiple blunt trauma: diagnostic and therapeutic difficulties. PMID- 16307987 TI - Stem cell transplantation for ischemic cardiomyopathy: hope or hype? PMID- 16307988 TI - Does the internal thoracic artery graft have self-reparative ability? PMID- 16307989 TI - Pulmonary atresia with ventricular septal defects and major aortopulmonary collateral arteries: unifocalization brings no long-term benefits. AB - OBJECTIVES: We sought to evaluate the contribution of unifocalization procedures in the management of patients with pulmonary atresia, ventricular septal defect, and major aortopulmonary collateral arteries. METHODS: From 1975 through 1995, 82 consecutive patients were entered in a multistage approach and had 189 sternotomies and thoracotomies to perform 119 shunts, 130 major aortopulmonary collateral artery transplantations, and 76 major aortopulmonary collateral artery ligations. The serial angiographies and the follow-up of these patients were reviewed. RESULTS: The concurrent follow-up rate was 80%. The hospital mortality of the preliminary procedures was 4% (7/189). Fifty-three (65%) patients had a complete repair. The hospital mortality of the repair was 8% (4/53), and 9 late deaths occurred after repair, all of which were cardiac related. The overall survival of all patients to the age of 30 years was 58% +/- 7%. Survival 12 years after complete repair was 51% +/- 14%. On angiography, central shunts promoted growth of central pulmonary arteries in all cases (29 patients). Sixty unifocalized major aortopulmonary collateral arteries were identified in 31 patients. After a mean of 3.2 +/- 4 years, 26 thrombosed, and 12 presented with a stenosis of greater than 50%. Serial measurements of 29 major aortopulmonary collateral arteries showed no signs of growth (P = .25). CONCLUSION: Long-term survival into adulthood of patients with pulmonary atresia, ventricular septal defect, and major aortopulmonary collateral arteries has been achieved with a multistage approach. However, late survival depends exclusively on the growth of the native pulmonary circulation. The few unifocalized major aortopulmonary collateral arteries that did not thrombose failed to grow. PMID- 16307990 TI - Matching procedure to morphology improves outcomes in neonates with tricuspid atresia. AB - OBJECTIVE: This study was undertaken to characterize morphologic substrate of tricuspid atresia with ventriculoarterial concordance and discriminate management strategies that lead to successful definitive repair. METHODS: From 1999 to 2004, a total of 150 babies with type I tricuspid atresia were enrolled from first diagnosis at 26 institutions. Antegrade pulmonary blood flow was absent in 19%, restricted in 54%, and unrestricted in 28%. Competing-risk methodology determined the time-related prevalence and risk factors for death versus cavopulmonary anastomosis and subsequent death versus Fontan completion. RESULTS: Overall 5 year survival was 86%. Initial palliation included systemic-pulmonary arterial shunt in 64%, pulmonary artery banding in 11%, and cavopulmonary anastomosis in 24%. Median age at cavopulmonary anastomosis was 6 months, with 83% undergoing bidirectional Glenn shunt and 17% undergoing hemi-Fontan procedure. By the age of 2 years, 89% had cavopulmonary anastomosis, 6% were dead, and 4% remained alive without cavopulmonary anastomosis. Risk factors for death without cavopulmonary anastomosis included presence of mitral regurgitation (P = .03) and palliation with systemic-pulmonary arterial shunts not originating from the innominate artery (P = .04). Factors associated with decreased transition rate to cavopulmonary connection included patient variables (younger admission age to a participating institution, noncardiac anomalies) and procedural variables (larger systemic-pulmonary arterial shunt diameter, previous palliation). Of patients undergoing cavopulmonary anastomosis, 75% had undergone a Fontan operation within 3 years. CONCLUSION: Smaller shunt size and decreased pulmonary blood flow decrease mortality after initial palliation and increase the rate of successful transition to cavopulmonary anastomosis. Outcomes can be improved by placing smaller shunts from the innominate artery, especially in patients with any mitral regurgitation. PMID- 16307991 TI - Effect of congenital heart disease on neurodevelopmental outcomes within multiple gestation births. AB - OBJECTIVES: We sought to assess the effect of congenital heart disease requiring surgical intervention with cardiopulmonary bypass at 6 months of age or less on developmental outcomes and growth at 1 year of age while controlling for socioeconomic status, prematurity, home environment, and parental intelligence. METHODS: We performed within-family comparison of 11 multiple-gestation births in which one child had congenital heart disease. At 1 year of age, the Bayley Scales of Infant Development II were administered, and growth parameters were assessed. Paired comparisons were made by using fixed effects regression conditioned on family. RESULTS: The multiple-gestation subjects were mildly premature on average (mean gestational age, 35.4 +/- 3.0 weeks). At 1 year of age, children with congenital heart disease scored lower on the Mental Development Index (85.0 +/- 19.3 vs 93.9 +/- 16.0, P = .037) and the Psychomotor Development Index (76.6 +/- 16.9 vs 91.3 +/- 14.9, P = .015) on the Bayley Scales of Infant Development II than did their siblings without congenital heart disease. There were no differences between siblings in weight, height, or head circumference. CONCLUSIONS: The presence of congenital heart disease requiring surgical intervention with cardiopulmonary bypass at 6 months of age or less is associated with a deficit in developmental achievement at 1 year of age, as measured by using the Bayley Scales of Infant Development II. PMID- 16307992 TI - Fontan operation with a viable and growing conduit using pedicled autologous pericardial roll: serial changes in conduit geometry. AB - OBJECTIVE: We sought to examine midterm results of the Fontan operation with an extracardiac conduit using pedicled autologous pericardial roll, with special attention to angiographic evaluation of serial changes in conduit geometry. METHODS: Of 202 patients subjected to the Fontan operation since 1996, the conduit was used in 28 patients who had intact pericardium. We retrospectively reviewed a consecutive series of these patients, and serial changes in conduit geometry were accessed in 16 patients who underwent catheter examinations twice at 1.1 +/- 0.4 and 4.5 +/- 1.4 years postoperatively. RESULTS: There was one early and one late death. Except for a patient with apicocaval juxtaposition in whom a conduit occlusion developed caused by compression between the vertebral bodies and the ventricle, there were no important complications. Angiographically, conduit volume, estimated by using the MULTI-SLICE method, increased significantly (3490 +/- 2166 to 5426 +/- 3081 mm3, P < .001), whereas the volume per body weight remained unchanged. Conduit diameter increased significantly at both the inferior vena caval end (16.8 +/- 4.8 to 19.8 +/- 4.8 mm, P < .001) and the pulmonary artery end (11.9 +/- 3.8 to 14.2 +/- 4.2 mm, P < .001), whereas the diameter indexed to the normal right pulmonary artery remained unchanged. The cross-sectional area of the conduit increased in parallel with that of the normal right pulmonary artery. The ratio of the widest to the narrowest diameter of the conduit exhibited no significant change. CONCLUSIONS: Midterm results of the Fontan operation with pedicled autologous pericardial roll were favorable. Proportional increase of conduit size was demonstrated, with its shape preserved. This suggested a potential of the conduit to grow and that growth correlated with somatic development. PMID- 16307993 TI - Brain magnetic resonance imaging abnormalities after the Norwood procedure using regional cerebral perfusion. AB - OBJECTIVES: Neurologic deficits are common after the Norwood procedure for hypoplastic left heart syndrome. Because of the association of deep hypothermic circulatory arrest with adverse neurologic outcome, regional low-flow cerebral perfusion has been used to limit the period of intraoperative brain ischemia. To evaluate the effect of this technique on brain ischemia, we performed serial brain magnetic resonance imaging in a cohort of infants before and after the Norwood operation using regional cerebral perfusion. METHODS: Twenty-two term neonates with hypoplastic left heart syndrome were studied with brain magnetic resonance imaging before and at a median of 9.5 days after the Norwood operation. Results were compared with preoperative, intraoperative, and postoperative risk factors to identify predictors of neurologic injury. RESULTS: Preoperative magnetic resonance imaging (n = 22) demonstrated ischemic lesions in 23% of patients. Postoperative magnetic resonance imaging (n = 15) demonstrated new or worsened ischemic lesions in 73% of patients, with periventricular leukomalacia and focal ischemic lesions occurring most commonly. Prolonged low postoperative cerebral oximetry (<45% for >180 minutes) was associated with the development of new or worsened ischemia on postoperative magnetic resonance imaging (P = .029). CONCLUSIONS: Ischemic lesions occur commonly in neonates with hypoplastic left heart syndrome before surgical intervention. Despite the adoption of regional cerebral perfusion, postoperative cerebral ischemic lesions are frequent, occurring in the majority of infants after the Norwood operation. Long-term follow-up is necessary to assess the functional effect of these lesions. PMID- 16307994 TI - Ubiquitin fusion degradation 1-like gene dysregulation in bicuspid aortic valve. AB - OBJECTIVE: Bicuspid aortic valve, the most common congenital cardiac malformation, is caused by fusion of valve cushions at the onset of valvulogenesis. Although its exact pathogenesis is still unclear, a genetic basis is appearing more and more likely. Search for a potential candidate gene by reviewing semilunar valve morphogenesis led us to the ubiquitin fusion degradation 1-like gene (UFD1L), which is highly expressed in the cardiac outflow tract during embryogenesis. METHODS: Aortic valves were collected during surgery from 39 patients with bicuspid aortic valve (mean age 56.8 +/- 18.1 years) and from 38 patients with tricuspid aortic valve (mean age 61.7 +/- 16.1 years). Fluorescence in situ hybridization was performed for detection of microdeletion, quantitative reverse transcriptase-polymerase chain reaction to measure gene expression, and Western blotting to analyze the amount of UFD1L gene product. RESULTS: No microdeletion was found in either group in the critical region of chromosome 22 containing the UFD1L gene. UFD1L gene expression, however, was significantly reduced in bicuspid aortic valve samples (median 787-fold) relative to tricuspid aortic valve samples (median 10,887-fold, P = .001). The amount of UFD1L gene product was also significantly diminished in bicuspid aortic valve samples (3.9 +/- 2.6 vs 8.4 +/- 4.8 optical density units, P < .05). CONCLUSION: Bicuspid aortic valve was associated with downregulation of UFD1L gene expression, supporting the hypothesis that bicuspid aortic valve is a genetic disorder, with the UFD1L gene as a potential candidate gene. PMID- 16307995 TI - Simultaneous management of congenital tracheal stenosis and cardiac anomalies in infants. AB - OBJECTIVE: The present article aims to describe our experience with patients who underwent simultaneous repair of congenital tracheal stenosis and cardiac anomalies. METHODS: Between January 2000 and December 2003, 9 infants underwent simultaneous surgical repair of a congenital tracheal stenosis and congenital heart disease. The intraoperative findings revealed localized tracheal stenosis in 3 patients. The funnel-type tracheal stenosis was present in 6 patients. Associated cardiac anomalies included ostium secundum atrial septal defect in 5 patients and ventricular septal defect in 2 patients, pulmonary artery sling in 4 patients, patent ductus arteriosus in 6 patients, atrioventricular septal defect in 1 patient, aortic arch hypoplasia in 1 patient, coarctation of the aorta in 1 patient, and partial anomalous pulmonary venous connection in 2 patients, one of them with "scimitar syndrome." Tracheal origin of the right upper lobe was diagnosed in 2 of the patients. A right aberrant subclavian artery (lusoria) was found in one patient. All patients were operated on through a median sternotomy and with cardiopulmonary bypass. Tracheal resection with direct end-to-end anastomosis was performed in all cases. RESULTS: There was no operative mortality. One patient died 6 weeks postoperatively. Eight patients were extubated between the 14th and 30th postoperative days under bronchoscopic monitoring. The extubation was performed after a stepwise respirator-weaning program. Postoperative endoscopic examination showed adequate airway dimensions and patency in every case. The midterm results after a mean follow-up of 37 months (range, 16-58 months) of the entire group demonstrate a stabile and complication-free clinical outcome. CONCLUSIONS: We advocate our current strategy for infants with congenital tracheal stenosis: resection with end-to-end anastomosis and simultaneous repair of associated intracardiac anomalies. PMID- 16307996 TI - The roles of chronic pressure and volume overload states in induction of arrhythmias: an animal model of physiologic sequelae after repair of tetralogy of Fallot. AB - OBJECTIVE: Sudden death occurs in as many as 8% of patients after repair of tetralogy of Fallot and has been attributed to arrhythmias. The purpose of this study was to establish an animal model to evaluate the individual contribution of different physiologic sequelae after tetralogy of Fallot repair in the development of late-onset arrhythmias. METHODS: Forty-nine piglets were divided into 5 groups: (1) pulmonary artery band; (2) pulmonary valvotomy; (3) pulmonary artery band plus pulmonary valvotomy; (4) infundibular scar; and (5) age-matched control animals. Baseline and follow-up electrocardiograms were obtained and recorded, as well as changes in QRS duration. A total of 45 animals underwent hemodynamic evaluation and programmed electrical stimulation at 5.6 months postoperatively. RESULTS: Sustained ventricular tachyarrhythmias (ventricular tachycardia/ventricular fibrillation) were induced in 31.1%, and atrial arrhythmias were induced in 33.3%. The pulmonary valvotomy group was 30 times more likely to evidence arrhythmias than control animals for sustained ventricular tachycardia/ventricular fibrillation, as well as atrial arrhythmias (P = .01). The pulmonary artery band group was 15 times more likely to evidence atrial arrhythmias than control animals (P = .02). Prolonged QRS duration was predictive of inducibility of both atrial arrhythmias (P < .01) and sustained ventricular tachycardia/ventricular fibrillation (P = .01). Mean right atrial (P = .01) and capillary wedge (P = .01) pressures predicted atrial arrhythmia inducibility. Right ventricular end-diastolic pressure predicted atrial arrhythmia (P= .01) and sustained ventricular tachycardia/ventricular fibrillation inducibility (P = .05). Right ventricular systolic pressure did not predict inducibility of either atrial arrhythmias (P = .10) or sustained ventricular tachycardia/ventricular fibrillation (P = .94). CONCLUSIONS: Chronic right ventricular volume overload resulted in an increased incidence of inducible ventricular and atrial arrhythmias. PMID- 16307997 TI - Ablation of atrial fibrillation and esophageal injury: effects of energy source and ablation technique. AB - OBJECTIVE: The aim of this study was the evaluation of histologic changes induced on the esophagus by surgical ablation therapy for atrial fibrillation. METHODS: Experiments were performed on 39 sheep. Circular lesions were created endocardially or epicardially in the left atrium and at the pulmonary veins by using different energy sources: cryoablation, microwave, laser, and unipolar or bipolar radiofrequency. Temperatures inside the esophagus were measured, and esophageal tissue was investigated macroscopically and histopathologically. RESULTS: Esophageal damage was seen histologically in 24 of 39 cases. The epithelium was intact in all cases. Unipolar radiofrequency induced the most intensive esophageal lesions in 4 of 6 cases. The affected areas were small (1.56 3.01 mm) but reached deep into the tissue. Endocardial cryoablation resulted in wider lesions (2.01-8.54 mm), which were intensive in only 2 of 6 cases. Epicardial cryoablation and bipolar radiofrequency induced wide (1.11-6.8 mm) but mainly mild alterations. Endocardial and epicardial microwave energy affected the esophagus in single cases, and lesions were small (0.97-2.81 mm). Only in 1 case did laser energy induce a moderate alteration (5.30 mm) of the esophageal wall. CONCLUSIONS: Esophageal alterations were found in numerous cases. However, marked lesions were especially induced by endocardial unipolar radiofrequency and cryoablation. PMID- 16307998 TI - Prediction of cardiac function after weaning from ventricular assist devices. AB - OBJECTIVE: We analyzed the factors influencing the cardiac function of patients after weaning from ventricular assist devices and established a prognostic index. METHOD: A multivariate analysis was performed using the Cox proportional hazards model of the clinical parameters of 28 patients with end-stage heart failure before implantation of a ventricular assist device and directly before weaning from the device. RESULTS: After weaning from the ventricular assist devices, 14 of the 28 investigated patients showed stable cardiac function, and 12 had recurrent heart failure, but only 1 of them died before transplantation or implantation of another ventricular assist device, and 2 patients died of causes unrelated to heart failure. In addition to left ventricular ejection fraction, the duration of symptomatic heart failure and left ventricular intracavitary dimensions in diastole measured before ventricular assist devices weaning were the major factors influencing cardiac function after weaning. In the group of patients with heart failure duration less than 3 years, only 2 of 13 cases were recurrent, but in the group with heart failure duration more than 3 years, 10 of 15 cases were recurrent. In 10 patients with 40 to 50 mm left ventricular intracavitary dimensions in diastole before weaning, only 1 case was recurrent. In 10 patients with 51 to 55 mm left ventricular intracavitary dimensions in diastole, 3 cases were recurrent. In the group with left ventricular intracavitary dimensions in diastole more than 56 mm, all 8 cases were recurrent. A prognostic index was calculated using the following formula: prognostic index = -10.10 + 0.208 (heart failure duration in years) + 0.173 (pre-explantation left ventricular intracavitary dimensions in diastole in millimeters). In the group with a prognostic index less than 0, only 2 of 16 cases were recurrent, and the rate of stable cardiac function in the 51st month after weaning was 83.6%. In the group with a prognostic index greater than 0, 10 of 12 cases were recurrent, and the rate of stable function in the 57th month was 0.0%. CONCLUSION: In patients with off-pump left ventricular ejection fraction greater than 40%, the duration of symptomatic heart failure and the left ventricular intracavitary dimensions in diastole measured before weaning yield a useful index to predict long-term cardiac function after weaning from ventricular assist devices. PMID- 16307999 TI - The role of tissue factor and P-selectin in the procoagulant response that occurs in the first month after on-pump and off-pump coronary artery bypass grafting. AB - BACKGROUND: It has been previously shown that a persistent (up to 1 month) prothrombotic status occurs after coronary bypass surgery performed both on pump and off pump. To assess the pathways involved in the occurrence of postoperative prothrombotic state, in this study we evaluated plasma, monocyte-bound, and platelet-bound tissue factor expression, as well as platelet and soluble P selectin expression, up to 1 month after off-pump and on-pump coronary artery bypass grafting. METHODS: Thirty patient candidates for coronary surgery were randomized to undergo off-pump coronary artery bypass grafting (n = 15) or on pump coronary artery bypass grafting (n = 15). Blood samples were collected before the intervention, after protamine administration, and 4, 8, and 30 days after surgical intervention. RESULTS: Plasma tissue factor levels were significantly higher than baseline both in the on-pump coronary artery bypass grafting group (from protamine administration up to 4 postoperative days) and in the off-pump coronary artery bypass grafting group (at 4 postoperative days), with no differences between groups. Basal and lipopolysaccharide-stimulated monocyte tissue factor expression, as well as basal and adenosine diphosphate stimulated platelet tissue factor expression, did not show significant variations over time and were similar in the on-pump and off-pump coronary artery bypass grafting groups throughout the course of the study. Platelet expression of P selectin, both basal and after adenosine diphosphate stimulation, did not significantly change over time and was not different in the on-pump and off-pump coronary artery bypass grafting groups. Soluble P-selectin levels in plasma were significantly higher in patients receiving on-pump coronary artery bypass grafting only at the time point after protamine administration, whereas this variable behaved similarly in the on-pump and off-pump coronary artery bypass grafting groups for the whole postoperative period. CONCLUSIONS: The postoperative tissue factor and P-selectin expression did not differ between the on-pump and off-pump coronary artery bypass grafting groups. The distinct increase of plasma tissue factor occurring after both surgical procedures might represent a mechanism that might explain, in part, the early postoperative prothrombotic state occurring after on-pump and off-pump coronary artery bypass grafting. PMID- 16308000 TI - Preoperative anti-PF4/heparin antibody level predicts adverse outcome after cardiac surgery. AB - OBJECTIVE: Preexisting serum antibodies to heparin/platelet factor 4 complexes may predispose adult cardiac surgical patients to increased perioperative morbidity and mortality. We sought to determine the association between preoperative serum antibodies directed against platelet factor 4/heparin complexes and major complications (in-hospital death or length of stay >10 days) in adult cardiac surgical patients. METHODS: In a prospective observational study of 466 patients undergoing elective coronary artery bypass grafting, valvular heart surgery, or both, preoperative serum was assayed for anti-platelet factor 4/heparin antibody by using a commercially available enzyme-linked immunosorbent assay (Asserachrom HPIA). Known preoperative risk factors were assessed, and patients were assigned a risk score by using the validated method of Parsonnet and colleagues. RESULTS: Major complications (death or postoperative hospitalization >10 days) occurred in 108 patients (23%). Overall, 59 (13%) patients had a positive preoperative anti-platelet factor 4/heparin antibody screen (upper limit of normal is 0.5 optical density units). A positive assay result independently predicted an increased risk of major complications (P = .0284; odds ratio, 1.98; 95% confidence interval, 1.06-3.62) over and above the effect of the Parsonnet risk score (P < .001; odds ratio, 1.07; 95% confidence interval, 1.05-1.10). The level of preoperative anti-platelet factor 4/heparin antibody was also significantly associated with major complications (P = .036; odds ratio, 1.31; 95% confidence interval, 1.02-1.68) independently of the Parsonnet risk score. No association (P > .75) existed between the Parsonnet risk score and preoperative anti-platelet factor 4/heparin antibody level. CONCLUSIONS: Serum antibodies directed against platelet factor 4/heparin complexes are prevalent in the adult patient population undergoing cardiac surgery. The presence of these antibodies before surgery is an independent predictor for death or prolonged hospitalization after adult cardiac surgery. PMID- 16308001 TI - The induction of heme oxygenase 1 decreases contractility in human internal thoracic artery and radial artery grafts. AB - OBJECTIVE: Spasm remains a potential problem encountered during the use of arterial grafts in coronary artery bypass surgery. Heme oxygenase plays a role in the control of arterial vasoreactivity. Heme oxygenase exists in 2 constitutive isoforms (heme oxygenase 2 and 3) and an inducible isoform (heme oxygenase 1). The aim of our study was to induce heme oxygenase 1 by using hemin in human internal thoracic and radial arteries and to evaluate the effect of this induction on the contractility of these arterial grafts. METHODS: Segments of human arterial grafts obtained from patients undergoing isolated coronary artery bypass surgery were incubated in organ chambers for 4 hours in the presence of 10(-4) mol/L hemin. Concentration-response curves to norepinephrine were obtained in control and hemin-treated arterial rings. Heme oxygenase 1 expression was evaluated by using enzyme-linked immunosorbent assays and immunohistochemical staining. RESULTS: The contractility of the arterial rings to norepinephrine was significantly reduced after incubation with hemin. Zinc protoporphyrin (an inhibitor of heme oxygenase) reversed the effect of hemin, whereas the inhibitor of nitric oxide synthase had no effect. The inhibitor of soluble guanylate cyclase blocked the decrease in contractility induced by hemin. Immunohistochemical staining revealed a large expression of heme oxygenase 1 in all vascular layers of hemin-treated internal thoracic artery and radial artery rings. Enzyme-linked immunosorbent assay studies showed a significant increase in heme oxygenase 1 levels in hemin-treated internal thoracic artery and radial artery rings. CONCLUSION: Hemin caused in vitro induction of heme oxygenase 1 in human internal thoracic artery and radial artery grafts. This induction resulted in a reduced contractility to norepinephrine, partially through the cyclic guanosine monophosphate-dependent pathway. This effect was independent from nitric oxide synthesis. PMID- 16308002 TI - Cerebral microembolization during off-pump coronary artery bypass surgery with the Symmetry aortic connector device. AB - OBJECTIVE: The use of aortic connector systems for proximal vein grafts in off pump coronary artery bypass grafting might minimize aortic manipulation by eliminating the need for partial aortic clamping. The objective of this study was to asses whether use of a Symmetry connector (St Jude Medical, Inc, St Paul, Minn) reduced intraoperative cerebral embolization. METHODS: Thirty-two consecutive patients underwent off-pump coronary artery bypass grafting. Sixteen patients received at least one mechanical proximal vein graft anastomosis with a Symmetry aortic connector system. Sixteen patients representing the control group underwent operations with standard suturing techniques using partial aortic clamping. During surgical intervention, all patients were monitored continuously with multifrequency transcranial Doppler scanning, which detected and differentiated cerebral emboli. RESULTS: There were significantly more cerebral emboli in the Symmetry group (median, 36) compared with the control group (median, 11; P = .027). This was due to a higher number of gaseous emboli in the Symmetry group than in the control group (median, 27 vs 8; P = .014), whereas there was no significant difference regarding the number of solid emboli (median, 7 vs 3; P = .139). CONCLUSION: Use of a Symmetry connector system during proximal vein graft anastomosis increased the number of emboli to the brain compared with a standard technique in coronary bypass surgery without cardiopulmonary bypass. PMID- 16308003 TI - Experimental study on the protective effects of edaravone against ischemic spinal cord injury. AB - OBJECTIVE: Reactive free radical species are thought to be involved in ischemic spinal cord injury. We investigated the effects of edaravone (Mitsubishi Pharma Co, Tokyo, Japan), a free radical scavenger, on spinal ischemia-reperfusion injury in a rabbit model. We also sought to estimate free radicals in the spinal cord using the microdialysis method. METHODS: Spinal cord ischemia was induced in New Zealand White rabbits. The animals were then divided into 4 groups. In the first experiment, which was carried out in group A (non-edaravone treated) and group B (edaravone treated), we assessed neurologic function and evaluated spinal cord histopathology. In the second experiment, which was performed in group C (non-edaravone treated) and group D (edaravone treated), we sequentially estimated the level of free radical species in the spinal cord with the microdialysis method. RESULTS: In the first experiment group B showed better neurologic function than group A. The number of viable neurons in the spinal cord gray matter was also higher in group B than in group A. The second experiment revealed that the level of free radical species was lower in group D at 75, 90, and 150 minutes after the beginning of reperfusion compared with levels seen in group C. The appearance of free radical species in the latter group was found to have a biphasic pattern, with peaks at 75 and 150 minutes after the beginning of reperfusion. CONCLUSION: Edaravone exerted a significant protective effect on the spinal cord against ischemia-reperfusion injury by suppressing the level of free radical species, which was demonstrated with the microdialysis method. PMID- 16308004 TI - A physiologic clinical study of achalasia: should Dor fundoplication be added to Heller myotomy? AB - OBJECTIVE: Myotomy for achalasia disrupts the lower esophageal sphincter, improving emptying at the expense of reflux. We hypothesized that surgical palliation of achalasia requires balancing desirable improvement in esophageal emptying with undesirable production of gastroesophageal reflux. Therefore, we objectively studied the physiologic effects of adding Dor fundoplication to Heller myotomy. METHODS: From December 1996 to June 2004, 149 patients underwent Heller myotomy; 88 (59%) had additional Dor fundoplication. The adequacy of myotomy was assessed by premyotomy to postmyotomy change in lower esophageal sphincter pressures, esophageal emptying by change in timed barium esophagram, and gastroesophageal reflux by postoperative 24-hour pH monitoring. RESULTS: For adequacy of myotomy, postmyotomy resting lower esophageal sphincter pressure was higher with (median, 18 mm Hg) than without (median, 13 mm Hg) Dor fundoplication (P = .002), as was residual lower esophageal sphincter pressure (median, 4.6 vs 1.8 mm Hg; P = .01). For esophageal emptying, postmyotomy barium height and width were similar with or without Dor fundoplication (P > .1). For gastroesophageal reflux, percentage of upright time with a pH of less than 4 was lower with (median, 0.4%) than without (median, 2.9%) Dor fundoplication (P = .005), and percentage of supine time with a pH of less than 4 was lower with (median, 0%) than without (median, 5.8%) Dor fundoplication (P = .007). CONCLUSIONS: The addition of Dor fundoplication reduces the adequacy of myotomy without impairing emptying and reduces reflux. Heller myotomy and Dor fundoplication balance emptying and reflux and therefore should be the surgical treatment of choice for achalasia. PMID- 16308005 TI - The evolution of treatment outcomes for resected stage IIIA non-small cell lung cancer over 16 years at a single institution. AB - OBJECTIVES: The effect of multimodality treatment including surgical intervention, chemotherapy, and radiation for potentially resectable stage IIIA non-small cell lung cancer in a practice setting remains to be defined. To determine which treatment factors are associated with improved survival, we evaluated outcomes for these patients at our institution over a 16-year period. METHODS: We surveyed our institutional pathology database from 1986 through 2001 for patients with resected pathologic stage IIIA (N2) non-small cell lung cancer. Three hundred fifty-three patients were confirmed to have appropriate pathologic staging and attempted complete resection. These patients were assessed by means of univariate and multivariable analysis for factors associated with long-term survival. Stage migration was estimated by using a classification based on nodal station involvement. RESULTS: Median potential follow-up was 132 months. During the study period, 3- and 5-year survival increased; preoperative staging improved, relatively more lobectomies and fewer pneumonectomies were performed, and multimodality treatment was used more frequently. The number of positive N2 nodal stations did not change over time (P = .14). Surgical intervention alone resulted in 3-year survival of 30%, and perioperative chemotherapy, radiation, or both increased 3-year survival to 38% (P = .004). Multivariable analysis showed that male sex (hazard ratio, 1.44; 95% confidence interval, 1.13-1.84; P = .003), more than 2 positive mediastinal nodal stations (hazard ratio, 1.73; 95% confidence interval, 1.16-2.57; P = .007), R1 or R2 resection (hazard ratio, 1.72; 95% confidence interval, 1.22-2.41; P = .002), lower or middle lobe tumor location (hazard ratio, 1.63; 95% confidence interval, 1.28-2.08; P < .001), and surgical intervention alone (hazard ratio, 1.59; 95% confidence interval, 1.23 2.04; P < .001) were independent predictors of poor survival. CONCLUSIONS: The use of multimodality therapy appears to contribute to improved outcomes over time in patients with resected stage IIIA (N2) non-small cell lung cancer. PMID- 16308006 TI - Positron emission tomographic scanning in the diagnosis and staging of non-small cell lung cancer 2 cm in size or less. AB - OBJECTIVE: Several studies have suggested that positron emission tomography is more accurate than computed tomography for the staging of non-small cell lung cancer and can reduce the rate of unnecessary thoracotomy in patients with potentially resectable disease. However, there are few data on the utility of positron emission tomography in the diagnosis of patients with tumors of 2 cm or less in size. METHODS: Patients with cT1/cT2 tumors of 2 cm or less in size were retrospectively reviewed. All had a computed tomographic scan, as well as a positron emission tomographic scan on a dedicated scanner, with a standard uptake value reported. A standard uptake value of 2.5 g/mL or greater was considered positive. The results of computed tomography and positron emission tomography were correlated with pathologic results after either resection (n = 60) or mediastinoscopy (n = 4). RESULTS: Sixty-four patients (38 women; mean age, 66 years) had a mean tumor size of 1.4 cm (range, 0.7-2.0 cm). Forty-three patients had adenocarcinoma, 13 had adenocarcinoma-bronchioloalveolar carcinoma, 5 had squamous cell carcinoma, and 3 had other tumor types. Twenty-nine (45%) tumors had negative positron emission tomographic results. Both tumor size (>1 cm vs < or =1 cm) and cell type (adenocarcinoma-bronchioloalveolar carcinoma vs all other cell types) were significant predictors of positron emission tomography uptake in the primary tumor (P = .05 and .01, respectively). Nodal metastases were detected pathologically in 11 (17%) patients (5 N1 and 6 N2). Positron emission tomographic sensitivity and specificity for nodal metastases were only 45% and 89%, respectively. There was no statistically demonstrable survival difference between positron emission tomography-positive and positron emission tomography negative tumors (3-year survival of 87% vs 100%, respectively). CONCLUSION: Positron emission tomographic scanning has no demonstrable benefit in the diagnosis, staging, or prognosis of patients with tumors of 2 cm or less in size. PMID- 16308007 TI - Does lobectomy for lung cancer in patients with chronic obstructive pulmonary disease affect lung function? A multicenter national study. AB - OBJECTIVE: The purpose of this study was to evaluate the effect of lobectomy on pulmonary function in patients with chronic obstructive pulmonary disease. METHODS: One hundred thirty-seven patients were analyzed; 49 had normal pulmonary function tests, and 88 had chronic obstructive pulmonary disease. Different functional parameter groups were identified: obstructive (forced expiratory volume in 1 second [FEV1], forced expiratory volume in 1 second/forced vital capacity [FEV1/FVC], and chronic obstructive pulmonary disease index), hyperinflation (residual volume and functional residual capacity), and diffusion (transfer factor of the lung for carbon monoxide). Also, the ratio between observed and predicted postoperative FEV(1) was calculated. RESULTS: In patients with preoperative FEV1 greater than 80% of predicted, postoperative FEV1/FVC slightly but not significantly decreased, and postoperative FEV1 significantly decreased. In patients with preoperative FEV1 less than 65%, postoperative FEV1 and FEV1/FVC significantly increased. In patients with preoperative FEV1/FVC greater than 70%, postoperative FEV1 and FEV1/FVC significantly decreased. In patients with preoperative FEV1/FVC less than 70%, postoperative FEV1/FVC increased, and FEV1 remained unchanged. In patients with a chronic obstructive pulmonary disease index greater than 1.5, postoperative FEV1 and FEV1/FVC significantly decreased, whereas in patients with a chronic obstructive pulmonary disease index less than 1.5, postoperative FEV1/FVC significantly increased and FEV1 remained unchanged. In patients with residual volume and functional residual capacity greater than 115% and transfer factor of the lung for carbon monoxide less than 80% of predicted, postoperative FEV1 diminished less (not significant) compared with patients who had residual volume and functional residual capacity less than 115% (P = .0001). Observed postoperative/predicted postoperative FEV1 was higher if FEV1/FVC was less than 55% (1.46), if FEV1 was less than 80% of predicted (1.21), or if the chronic obstructive pulmonary disease index was less than 1.5 (1.17). CONCLUSIONS: Patients with mild to severe chronic obstructive pulmonary disease could have a better late preservation of pulmonary function after lobectomy than healthy patients. PMID- 16308008 TI - Pretreatment with hyperbaric oxygen and its effect on neuropsychometric dysfunction and systemic inflammatory response after cardiopulmonary bypass: a prospective randomized double-blind trial. AB - OBJECTIVE: Animal studies have shown that pretreatment with hyperbaric oxygen can induce central nervous system ischemic tolerance and also modulate the inflammatory response. We evaluated this therapy in patients undergoing cardiopulmonary bypass. METHODS: Sixty-four patients were prospectively randomized to group A (n = 31; atmospheric air, 1.5 atmospheres absolute) or group B (n = 33; hyperbaric oxygen, 2.4 atmospheres absolute) before on-pump coronary artery bypass grafting. Age, sex, body mass index, diabetes, hypertension, smoking, coronary disease severity, left ventricular function, Parsonnet score, Euroscore, bypass time, myocardial ischemia time, and number of grafts were comparable in both groups. Canadian Cardiovascular Society angina, New York Heart Association dyspnea, and previous myocardial infarction were significantly higher in group B. Inflammatory markers were analyzed before surgery and 2 and 24 hours after bypass. Neuropsychometric testing was performed 48 hours before surgery and 4 months after surgery and included trail making A and B, the Rey auditory verbal learning test, grooved peg board, information processing table A, and digit span forward and backward. Neuropsychometric dysfunction was defined as more than 1 SD deterioration in more than 2 neuropsychometric tests. Chi-square tests, Fisher tests, t tests, and analysis of variance were used as appropriate for statistical analysis. RESULTS: Group A had a significant postoperative increase in the inflammatory markers soluble E selectin, CD18, and heat shock protein 70. This was not observed in group B. Neuropsychometric dysfunction was also significantly higher in group A compared with group B. There was no difference in any other early postoperative clinical outcome. CONCLUSIONS: Our results seem to indicate that pretreatment with hyperbaric oxygen can reduce neuropsychometric dysfunction and also modulate the inflammatory response after cardiopulmonary bypass. However, further multicenter randomized trials are needed to clinically evaluate this form of therapy. PMID- 16308009 TI - Surgical treatment for congestive heart failure with autologous adult stem cell transplantation: a prospective randomized study. AB - BACKGROUND: Autologous adult stem cell transplantation has been touted as the latest tool in regenerative medical therapy. Its potential for use in cardiovascular disease has only recently been recognized. A randomized study was conducted with a novel epicardial technique to deploy stem cells as an adjuvant to conventional revascularization therapy in patients with congestive heart failure. METHODS: After institutional review board and government approval, adult autologous stem cell transplantation (CD34+) was performed in patients with ischemic cardiomyopathy and an ejection fraction of less than 35% who were scheduled for primary off-pump coronary artery bypass grafting. Preoperatively, the patients underwent echocardiography, stress thallium imaging single photon emission computed tomography, and cardiac catheterization to identify ischemic regions of the heart and to guide in the selection of stem cell injection sites. The patients were prospectively randomized before the operative therapy was performed. Patient follow-up was 1, 3, and 6 months with echocardiography, single photon emission computed tomography, and angiography. RESULTS: There were 20 patients enrolled in the study. Ten patients had successful subepicardial transplantation of autologous stem cells into ischemic myocardium. The other 10 patients, the control group, only had off-pump coronary artery bypass grafting. There were 8 male and 2 female subjects in each group. The median number of grafts performed was 1 in both groups. On angiographic follow-up, all grafts were patent at 6 months. The ejection fractions of the off-pump coronary artery bypass grafting group versus the off-pump coronary artery bypass grafting plus stem cell transplantation group were as follows: preoperative, 30.7% +/- 2.5% versus 29.4% +/- 3.6%; 1 month, 36.4% +/- 2.6% versus 42.1% +/- 3.5%; 3 months, 36.5% +/- 3.0% versus 45.5% +/- 2.2%; and 6 months, 37.2% +/- 3.4% versus 46.1% +/- 1.9% (P < .001). There were no perioperative arrhythmias or neurologic or ischemic myocardial events in either group. CONCLUSIONS: Autologous stem cell transplantation led to significant improvement in cardiac function in patients undergoing off-pump coronary artery bypass grafting for ischemic cardiomyopathy. Further investigation is required to quantify the optimal timing and specific cellular effects of the therapy. PMID- 16308010 TI - A novel device for left atrial appendage exclusion. AB - OBJECTIVE: The left atrial appendage is a frequent source of thromboemboli in patients with atrial fibrillation. Exclusion or excision of the left atrial appendage may reduce the risk of stroke in patients with atrial fibrillation. We evaluated the ability of a novel device to exclude the left atrial appendage during early and intermediate follow-up periods in a canine model. METHODS: Eight mongrel dogs (mean weight 29.1 +/- 4.0 kg) were used in this study. The occlusion device, constructed from 2 stainless steel strips covered with a knit braided polyester fabric, was implanted at the base of the left atrial appendage through a left thoracotomy on a beating heart. Dogs were evaluated at 7 days (n = 2), 30 days (n = 2), and 90 days (n = 4) by epicardial echocardiography, left atrial angiography, histologic inspection, and gross pathology. RESULTS: Device implantation was performed without complications in all animals. Complete exclusion of the left atrial appendage from the circulation was confirmed acutely and chronically by echocardiographic and angiographic evaluations. There was no device migration or damage to adjacent structures. CONCLUSION: This novel device enables rapid, reliable, and safe exclusion of the left atrial appendage. The device provides a new therapeutic option for reducing the risk of stroke in patients with atrial fibrillation. PMID- 16308011 TI - The Cardica C-Port System: clinical and angiographic evaluation of a new device for automated, compliant distal anastomoses in coronary artery bypass grafting surgery--a multicenter prospective clinical trial. AB - OBJECTIVES: The C-Port System (Cardica, Inc, Redwood City, Calif) integrates in one tool all functions necessary to enable rapid automated distal coronary anastomoses. The goal of this prospective, nonrandomized, and multicenter study is to determine the safety and efficacy of this novel anastomotic system. METHODS: Five centers enrolled 133 patients awaiting elective coronary artery bypass grafting surgery. Outcome variables were intraoperative device performance, incidence of device-related adverse events, predischarge and 6-month angiographic graft patency, and 12-month clinical outcome. Independent core laboratories performed qualitative and quantitative angiographic and computed tomographic assessments. RESULTS: The C-Port was used to perform a vein-to coronary anastomosis in 130 patients. Intraoperative conversion to a hand-sewn anastomosis was necessary in 11 patients because of inadequate target site preparation, inappropriate target vessel selection, or both. Inadequate blood flow related to poor runoff required conversion in 3 additional patients. Three patients died before discharge of causes unrelated to the device. At discharge, 113 patients had a C-Port implant in place, and 104 C-Port anastomoses were studied by means of angiography, resulting in 100 FitzGibbon A, 3 FitzGibbon B, and 1 FitzGibbon 0 classifications. At 6 months, one additional patient died of a device-unrelated cause, and 98 patients were evaluated by means of angiography (n = 89). Overall patency (FitzGibbon A) was 92.1%. Three C-Port anastomoses were rated FitzGibbon B, and 4 were rated FitzGibbon 0. At 12 months, 107 (98.2%) of 109 alive patients were followed up, without any reports of device-related major adverse cardiac events. CONCLUSIONS: The C-Port System allows for a rapid, reliable, and compliant distal anastomosis and yields favorable 6-month angiographic and 12-month clinical results when compared with published studies. PMID- 16308012 TI - The Cox maze procedure in mitral valve disease: predictors of recurrent atrial fibrillation. AB - OBJECTIVES: The Cox maze procedure is the gold standard for ablation of atrial fibrillation in patients undergoing mitral valve surgery, and new approaches to atrial fibrillation ablation must be compared with it. Therefore, we sought to determine the time-related prevalence of atrial fibrillation and its risk factors after combined Cox maze and mitral valve surgery. METHODS: From November 1991 through January 2004, 263 patients (mean left atrial diameter, 5.8 +/- 1.2 cm) underwent combined mitral valve surgery (repair in 71%) and a cut-and-sew Cox maze procedure for atrial fibrillation (permanent, 74%; persistent, 7%; paroxysmal, 16%). Rhythm documented on 2367 postoperative electrocardiograms was used to estimate the prevalence of atrial fibrillation across time. RESULTS: Hospital mortality was 1.9%. Postoperative atrial fibrillation prevalence peaked at 36% at 2 weeks, decreasing to 21% at 5 years. Risk factors for higher postoperative atrial fibrillation prevalence varied with time and included longer duration of preoperative atrial fibrillation (P = .003), larger left atrial diameter (P = .01), older age (P = .0002), and higher left ventricular mass index (P = .02). CONCLUSIONS: In some patients undergoing mitral valve surgery and a Cox maze procedure, atrial fibrillation recurs over time, mandating close, long term follow-up of heart rhythm. Earlier operation and left atrial size reduction should be considered to improve results in selected patients. PMID- 16308013 TI - Late regression of left internal thoracic artery graft stenosis at the anastomotic site without intervention therapy. AB - OBJECTIVE: Intervention therapy has been recently performed on the left internal thoracic artery graft stenosis. The purpose of this study was to evaluate the natural course of the left internal thoracic artery graft stenosis at the anastomotic site and clarify whether intervention therapy should be performed early after surgery. METHODS: We investigated early angiographic results of the left internal thoracic artery graft in 343 consecutive patients who underwent coronary bypass surgery. In 100 of 343 patients who underwent follow-up angiography, the graft diameter and percentage diameter stenosis at the anastomotic site were compared between early postoperative and follow-up angiography. None of these patients underwent intervention therapy on the left internal thoracic artery graft. RESULTS: Of 343 patients, 46 showed 50% or greater diameter stenosis, and 20 showed 70% or greater diameter stenosis at the anastomotic site. In the 100 patients with follow-up angiography, the graft diameter significantly increased (1.8 +/- 0.4 vs 2.1 +/- 0.5 mm, P < .0001) at follow-up angiography. The percentage diameter stenosis significantly decreased (69% +/- 13% vs 35% +/- 20%, P < .0001) at follow-up angiography in the patients with 50% or greater diameter stenosis at early postoperative angiography. Regression of left internal thoracic artery graft stenosis was detected in most patients with 70% or greater diameter stenosis. CONCLUSIONS: Our study demonstrated that left internal thoracic artery graft stenosis at the anastomotic site at early postoperative angiography might improve without intervention therapy. We should consider the natural course of the left internal thoracic artery graft stenosis in determining the indication of intervention therapy early after surgery. PMID- 16308014 TI - Carpentier-Edwards PERIMOUNT Magna bioprosthesis: a stented valve with stentless performance? AB - OBJECTIVE: We designed this study to evaluate the early hemodynamic performance of the recently introduced Carpentier-Edwards PERIMOUNT Magna bioprosthesis (Edwards Lifesciences, Irvine, Calif) and compare it with those of the conventional Carpentier-Edwards PERIMOUNT stented bioprosthesis (Edwards Lifesciences) and Edwards Prima Plus porcine stentless bioprosthesis (Edwards Lifesciences). METHODS: Sixty-three patients (>70 years old) were enrolled in this prospective, randomized study. At operation, once the annulus had been measured, the best size suitable was assessed for each of the three valves before random assignment. Transthoracic echocardiography was performed before discharge to evaluate early postoperative hemodynamic performances of the different valves implanted. RESULTS: The best size suitable of Edwards Prima Plus (24.3 +/- 1.7 mm) was significantly superior to those of both the Carpentier-Edwards PERIMOUNT Magna (23.4 +/- 2.1 mm) and Carpentier-Edwards PERIMOUNT (22.4 +/- 1.8 mm). The best size suitable of the Carpentier-Edwards PERIMOUNT Magna, however, was significantly superior to that of the Carpentier-Edwards PERIMOUNT. Furthermore the best size suitable of the Carpentier-Edwards PERIMOUNT Magna was equal to the measured annulus in 55% of patients, as opposed to 25% for the Carpentier-Edwards PERIMOUNT (P < .001). Mean implanted labeled size of the Edwards Prima Plus was significantly higher than those of both the Carpentier-Edwards PERIMOUNT Magna and the Carpentier-Edwards PERIMOUNT (24.6 +/- 1.9 mm, 23.1 +/- 1.9 mm, and 22.5 +/- 1.8 mm, respectively). Early postoperative hemodynamic performance of the Carpentier-Edwards PERIMOUNT Magna, however, was superior to those of both the Edwards Prima Plus and the Carpentier-Edwards PERIMOUNT in both effective orifice area index (1.07 +/- 0.4 cm2/m2, 0.87 +/- 0.3 cm2/m2, and 0.80 +/- 0.2 cm2/m2, respectively) and mean peak gradient (20 +/- 6 mm Hg, 27 +/- 8 mm Hg, and 28 +/- 12 mm Hg, respectively). CONCLUSION: The improved design of the recently introduced third-generation stented bioprosthesis Carpentier-Edwards PERIMOUNT Magna allows implantation of a significantly bigger valve than with the old generation. Furthermore, the improved hemodynamic performance of the Carpentier Edwards PERIMOUNT Magna compares favorably with both the Carpentier-Edwards PERIMOUNT and the Edwards Prima Plus. PMID- 16308015 TI - Strut chordal-sparing mitral valve replacement preserves long-term left ventricular shape and function in pigs. AB - OBJECTIVE: Mitral valve replacement with preservation of the entire subvalvular apparatus entails superior postoperative left ventricular function compared with other techniques. However, this option is often not possible because of valve pathology. We hypothesized that preservation of only 4 mitral valve secondary ("strut") chordae would be functionally and geometrically equivalent to total valve preservation in the long-term setting. In a porcine mitral valve replacement model we investigated the long-term effects of 3 surgical techniques on left ventricular function and geometry: (1) total preservation of the native valve, (2) strut chordal preservation, and (3) total excision of the subvalvular apparatus. METHODS: Forty 60-kg pigs were randomized to 1 of the 3 techniques. Global and regional left ventricular function and dimensions were assessed with cardiovascular magnetic resonance and conductance catheter 90 days after mitral valve replacement. Groups were compared by multivariate analysis of variance. RESULTS: There was no overall difference between groups 1 and 2. Group 3 animals had (1) greater base-apex diastolic and systolic lengths, and smaller short-axis diameters, and (2) lower sphericity indices, and greater base-apex and short-axis fractional shortening than groups 1 and 2. Regional analysis showed slimming and elongation to occur primarily in the basal left ventricular segments. Left ventricular contractility and hemodynamic parameters did not differ between groups. CONCLUSIONS: Strut chordal preservation was equivalent to total valve preservation during mitral valve replacement, whereas total chordal resection caused significant left ventricular slimming with compensatory increases in fractional shortening. Therefore, to preserve left ventricular geometry, special attention must be paid to maintain the valvular-ventricular continuity through the strut chordae during mitral valve replacement. This concept may have important therapeutic implications for chordal-sparing mitral valve replacement. PMID- 16308016 TI - A Gbetagamma inhibitor reduces intimal hyperplasia in aortocoronary saphenous vein grafts. AB - OBJECTIVE: Approximately 50% of aortocoronary saphenous vein grafts are occluded 10 years after coronary revascularization surgery. Intimal hyperplasia, a critical component in saphenous vein graft failure, is defined by vascular smooth muscle cell proliferation, which is mediated in part by betagamma subunits of heterotrimeric G proteins (G(betagamma)) and downstream effectors such as mitogen activated protein kinases. A peptide consisting of the carboxyl-terminus of the beta-adrenergic receptor kinase (betaARKct) binds G(betagamma), thereby inhibiting G(betagamma) signaling. Utilizing a recombinant adenovirus containing the coding sequence for the betaARKct peptide (AdbetaARKct), this study investigates whether treatment of the vein graft with AdbetaARKct reduces intimal hyperplasia in a large animal model of aortocoronary saphenous vein graft intimal hyperplasia. METHODS: Twenty-seven dogs (27-32 kg) underwent aortocoronary bypass grafting to the left anterior descending artery using autologous saphenous vein. Vein grafts were treated with saline (n = 8), an empty adenovirus (n = 8), or AdbetaARKct (n = 8). A subset of dogs (n = 3) were sacrificed on postoperative day 7 and betaARKct expression confirmed by Northern blotting. RESULTS: Arteriograms performed on postoperative day 90 confirmed that saphenous vein grafts were patent. At postoperative day 90, AdbetaARKct-treated grafts demonstrated reduced intimal area compared to empty virus and saline treated animals (P < .05). Additionally, AdbetaARKct treatment of isolated vascular smooth muscle cells in vitro inhibited mitogen-activated protein kinase activation and decreased overall vascular smooth muscle cell proliferation. CONCLUSION: This study demonstrates that betaARKct expression in aortocoronary saphenous vein grafts reduces intimal hyperplasia and decreases vascular smooth muscle cell proliferation in vitro via inhibition of G(betagamma)-mediated mitogen-activated protein kinase activation. Modulation of G(betagamma) via betaARKct may represent a novel therapy to reduce intimal hyperplasia and saphenous vein graft failure. PMID- 16308017 TI - Preoperative cerebral ischemic lesions predict physical health status after on pump coronary artery bypass surgery. AB - BACKGROUND: Risk assessment is integral to patient selection and counseling before coronary artery revascularization. We studied the predictive ability of cerebral magnetic resonance imaging of preoperative and postoperative cerebral ischemic injury on self-reported physical and mental health at 3 months after coronary artery bypass surgery with or without use of cardiopulmonary bypass. METHODS: In a prospective clinical trial comparing on-pump and off-pump surgery, 120 patients responded to a questionnaire for self-report of angina (Canadian Cardiovascular Society scale) and physical and mental health status (Short Form 36) at baseline before preoperative cerebral magnetic resonance imaging. Preoperative sets of both magnetic resonance imaging and self-assessments were available for 103 (85.8%) patients. These patients were grouped according to classification of preoperative cerebral magnetic resonance imaging findings. Analysis of covariance determined the association of (1) preoperative magnetic resonance imaging status, (2) new postoperative cerebral lesions, and (3) actual use of cardiopulmonary bypass to physical and mental health. RESULTS: At 3 months after surgical intervention, 98 of 103 patients completed follow-up. The analysis revealed an interaction effect of preoperative cerebral ischemic injury and use of cardiopulmonary bypass on physical health (F = 9.07, P = .003) independent of age. No independent effects on health status were found of baseline magnetic resonance imaging or new cerebral lesions at 3 months. CONCLUSIONS: This study strongly suggests that the combination of preoperative cerebral ischemic injury and use of cardiopulmonary bypass can predict postoperative health status at 3 months. Cerebral magnetic resonance imaging might be a more specific indicator than age for preoperative assessment of vulnerability or resilience during rehabilitation after on-pump cardiac surgery. PMID- 16308018 TI - Surgical ventricular remodeling for multiterritory myocardial infarction: defining a new patient population. AB - OBJECTIVE: Because of limited medical and surgical options for patients with end stage congestive heart failure, we expanded the criteria for surgical ventricular remodeling to include patients with multiterritory myocardial infarction, a group historically considered high-risk candidates. We present our series of patients with multiterritory myocardial infarction who underwent surgical ventricular remodeling and propose a new patient population who may benefit from this procedure. METHODS: Data were analyzed for 51 consecutive patients undergoing surgical ventricular remodeling from January 2002 to June 2004, with 100% follow up. Three left ventricular vascular territories were defined: anteroapicoseptal (left anterior descending), lateral (circumflex), and inferior (right coronary artery). Infarction was assessed with magnetic resonance imaging and intraoperative findings. RESULTS: Multiterritory myocardial infarction was found in 64.7% of patients (33/51) undergoing surgical ventricular remodeling. Mean age was 61.6 +/- 11.1 years (range 40-81 years). Sixty-one percent (20/33) demonstrated evidence of myocardial infarction in all three territories. Five patients underwent concomitant mitral valve repair or replacement. Operative mortality was 6.1% (2/33) and did not differ from that of patients with single territory infarction (11.1%, P = .61). Surgical ventricular remodeling significantly improved left ventricular volumes and ejection fraction in patients with multiterritory myocardial infarction. Three patients required assist device implantation, and 2 patients required defibrillator placement. Sixty-nine percent of patients in preoperative New York Heart Association functional class III or IV (22/32) had improvement to class I or II at follow-up (P < .01). Cox regression analysis discriminated a preoperative left ventricular end-systolic volume index greater than 100 mL/m(2) as a significant risk factor for mortality (odds ratio 12.1, 95% confidence interval 1.27-114.51, P = .03). Thirty-month survival of patients with multiterritory myocardial infarction (73.5% +/- 8.3%) did not differ statistically from that of patients with single-territory infarction (n = 18). CONCLUSION: Surgical ventricular remodeling improves cardiac function and New York Heart Association functional status in patients with multiterritory myocardial infarction. Our initial results are promising and should prompt further studies to confirm our results and potentially expand the surgical ventricular remodeling inclusion criteria to include patients with multiterritory myocardial infarction. PMID- 16308019 TI - Radical surgical resection of a giant pleural metastasis of a malignant phyllodes tumor of the breast. PMID- 16308020 TI - Fibrovascular polyps of the esophagus. PMID- 16308021 TI - Cardiac tamponade: rare presentation of acquired pericardial-esophageal fistula. PMID- 16308022 TI - Aortic valve replacement for a case of anomalous origin of the left coronary artery from posterior sinus of Valsalva with intramural aortic course. PMID- 16308023 TI - Technical feasibility of Impella Recover 100 microaxial left ventricular assist device placement after biologic aortic valve replacement (21 mm) for postcardiotomy failure. PMID- 16308024 TI - One-stage intracardiac repair in combination with external stenting of the trachea and right bronchus for tetralogy of Fallot with an absent pulmonary valve and tracheobronchomalacia. PMID- 16308025 TI - A particular anatomic variation of the course of the left pulmonary artery. PMID- 16308026 TI - A modified elephant trunk technique: the 3-fold elephant trunk technique. PMID- 16308027 TI - Acute intrathoracic gastric volvulus after laparoscopic fundoplication: laparoscopic reduction and repair. PMID- 16308028 TI - Intravenous leiomyomatosis with extension to the heart: rare or underestimated? PMID- 16308029 TI - Diagnosis of a solitary cardiac metastasis from ocular melanoma. PMID- 16308030 TI - Adult Bochdalek hernia complicated with a perforated colon. PMID- 16308032 TI - Effect of surgical case volume on outcome after the Norwood procedure. PMID- 16308034 TI - Video-assisted thoracoscopic surgery for pulmonary tuberculosis. PMID- 16308037 TI - Happy holidays! PMID- 16308038 TI - Head for the hill. PMID- 16308039 TI - Nurse/victim: the fallacy of the divide. PMID- 16308040 TI - A descriptive study of the perceptions of workplace violence and safety strategies of nurses working in level I trauma centers. AB - INTRODUCTION: Workplace violence is a significant occupational hazard in health care. As the largest group of employees in health care, nurses are particularly vulnerable to workplace violence, with those who work in emergency departments being especially at risk. The purpose of this research was to study the phenomenon of workplace violence by interviewing emergency nurses who had experienced violence while on duty. METHOD: A descriptive study approached the issue of workplace violence from the perspective of 8 registered nurses from 2 level I trauma centers who volunteered to be interviewed. Cross-case comparison of the interview responses was used to analyze the data from verbatim transcripts. RESULTS: Emergency nurses identified specific experiences of violence at work. Inadequate safety measures and vulnerability were the 2 themes that were consistently verbalized through out the interviews. IMPLICATIONS FOR NURSING PRACTICE: The emergency nurses who were interviewed discussed their experiences with patients, family members, and others who exhibited violent and aggressive behavior. They identified safety measures that they believed were inadequate and discussed their feelings of vulnerability because of violent incidents at work. Further research with larger samples could confirm specific safety problems in emergency departments that must be addressed to provide a safer workplace for emergency nurses, their colleagues, and their patients. PMID- 16308041 TI - Emergency response to the Gulf Coast devastation byHurricanes Katrina and Rita: experiences and impressions. PMID- 16308042 TI - The creation of a behavioral health unit as part of the emergency department: one community hospital's two-year experience. PMID- 16308043 TI - A 4-year-old boy with pulmonary hemosiderosis and respiratory distress requiring use of a cuffed endotracheal tube. PMID- 16308044 TI - Allow natural death: a more humane approach to discussing end-of-life directives. PMID- 16308045 TI - A percutaneous coronary intervention kit and program and PCI kit: reducing door to-cath lab time. PMID- 16308046 TI - The aftermath of workplace violence: one person's account. PMID- 16308047 TI - An informal discussion of emergency nurses' current clinical practice: what's new and what works. PMID- 16308048 TI - Look-alike and sound-alike drugs: errors just waiting to happen. PMID- 16308049 TI - Motorcycle crash with multiple pelvic injuries. PMID- 16308050 TI - An emergency nurse goes to Washington: feeling legislative power at the US Capitol. PMID- 16308051 TI - The last full measure. PMID- 16308052 TI - A patient with an undetected evolving stroke: legal lessons learned. PMID- 16308054 TI - Understanding the assessment and treatment of caustic ingestions and the resulting burns. PMID- 16308055 TI - Epistaxis following an assault: practical considerations in stopping the bleeding. PMID- 16308056 TI - It takes more than string to fly a kite: 5-level acuity scales are effective, but education, clinical expertise, and compassion are still essential. PMID- 16308057 TI - Celebrating nursing: quilt begun by ED staff mushroomed to successful hospital wide project. PMID- 16308058 TI - Impact of emergency department volume on registered nurse time at the bedside. AB - STUDY OBJECTIVE: We determine how emergency department (ED) registered nurses (RNs) allocate their time between various tasks and describe how RN task distribution changes as a function of various measures of ED patient volume and patient acuity. METHODS: This is a 3-year direct observational study using a convenience sample of 63 8-hour nurse shifts. Four RN task categories were defined: (1) direct patient care, (2) indirect patient care, (3) non-RN care, and (4) personal time. Two measures of nurse workload were used, the patient-to-nurse ratio and the ED acuity index (ED acuity index=(Sigma reverse order triage scores/half hour)/total number of nurses staffing). Trained observers classified RN activity at 1-minute intervals during 8-hour shifts daily for 7 nonconsecutive 24-hour periods. RN staffing data, ED patient census, and patient triage scores were collected every half hour. Summary statistics, correlation tables, and regression analysis were used to establish relationships between RN task allocation, patients per nurse, and the ED acuity index. RESULTS: For the 63 nursing shifts studied, on average RNs spent 25.6% of their time performing direct patient care, 48.4% on indirect patient care, 6.8% on non-RN care, and 19.1% on personal time. Regardless of the number of patients per RN, approximately twice as much time is spent on indirect patient care as direct patient care. The correlation between the ED acuity index and the patient-to nurse ratio was 0.98. CONCLUSION: Regardless of workload, RNs spend the majority of their time performing indirect patient care. RNs spend little time performing tasks that could be performed by ancillary staff. The patient-to-nurse ratio performs just as well as a more complicated acuity index to measure the workload of RNs within an ED. PMID- 16308059 TI - Images in emergency medicine. Toddler's fracture. PMID- 16308060 TI - Impact of rapid entry and accelerated care at triage on reducing emergency department patient wait times, lengths of stay, and rate of left without being seen. AB - STUDY OBJECTIVE: Patients who leave before being seen by a physician represent a significant problem for many emergency departments (EDs). We sought to determine the effect of a new ED rapid entry and accelerated care at triage (REACT) process on the frequency of patients who leave before being seen. METHODS: We conducted a before-after intervention design to study the effect of REACT for ambulatory patients presenting to our urban academic center ED with a census of approximately 37,000. This process redesign included patient identification tracking, integrated computer interfaces to eliminate up-front registration tasks, immediate placement of patients in open ED beds, and physician-directed ancillary testing and care at triage when no ED beds were available. Outcome measures included the average monthly rate of patients who left before being seen during the 6 months before (pre-REACT) and 6 to 12 months after (post-REACT) its initiation. Other measures included average of mean monthly rates of wait times, ED length of stay, ED census, and admissions. RESULTS: There was a significant decrease in leave before being seen frequency from the pre-REACT to post-REACT periods (3.2% absolute decrease [95% confidence interval (CI) 1.9% to 4.6%]), despite an overall increase in ED census. Average mean monthly patient wait times decreased by 24 minutes [95% CI 10 to 38 minutes] after the initiation of REACT, as did overall ED length of stay by 31 minutes [95% CI 6 to 57 minutes]. CONCLUSION: The initiation of a rapid entry and accelerated care process significantly decreased patient leave before being seen rates, average wait times and length of stay, despite an overall increase in patient census. PMID- 16308062 TI - Images in emergency medicine. Malrotation with midgut volvulus. PMID- 16308063 TI - Team assignment system: expediting emergency department care. AB - STUDY OBJECTIVE: We designed and implemented an emergency department (ED) team assignment system, each team consisting of 1 emergency physician, 2 nurses, and usually 1 technician. Patients were assigned in rotation upon arrival to a specific team that was responsible for their care. We monitored the time from arrival to physician assessment, percentage of patients who left without being seen by a physician, and patient satisfaction before and after team assignment system implementation. METHODS: This study was done in a suburban community hospital with an annual ED census of approximately 39,000. Time to physician assessment was defined from the completion of the medical screening evaluation by an ED nurse at triage to initiation of emergency physician evaluation. Times were documented on the ED paper record and manually entered into a computerized registration by the clerical staff. Patients who left without being seen was reported as percentage of total ED visits. Patient satisfaction scores using a 5 point Likert scale to assess satisfaction with the emergency physician, ED staff courtesy, and coordination of care were gathered every 3 months from random mailings to a subset of patients. RESULTS: The 12-month ED census was 38,716 before team assignment system implementation and 39,301 afterwards. Complete time data were recorded for 34,152 (88.2%) and 32,537 (82.8%) of the patients, respectively. The mean time to physician assessment was 71.3+/-7.0 minutes before and 61.8+/-6.4 minutes after team assignment system implementation (absolute difference -9.5 minutes; 95% confidence interval [CI] -5.8 to -13.5 minutes). The percentage of patients seen by a physician within 1 hour was 56.3% before and 64.0% after team assignment system implementation (absolute difference 7.7%; 95% CI 5.1% to 10.3%). The percentage of patients who waited more than 3 hours for physician assessment was 17.8% before and 11.8% after team assignment system implementation (absolute difference -6.0%, 95% CI -4.0% to -8.1%). Before team assignment system, the left without being seen rate was 2.3% compared to 1.6% after team assignment system (absolute difference -0.8%; 95% CI -0.4% to -1.1%). Patient satisfaction reported as very good or excellent showed improvement in satisfaction with the physician (absolute increase 3.1%; 95% CI 1.0% to 5.3%), staff courtesy (absolute increase 4.5%; 95% CI 2.3% to 6.7%), and coordination of care (absolute increase 3.6%; 95% CI 0.8% to 6.4%). CONCLUSION: The implementation of a team assignment system in our ED was associated with reduced time to physician assessment, a reduced percentage of patients who left without being seen, and improved patient satisfaction. PMID- 16308065 TI - Accuracy of ECG interpretation in the pediatric emergency department. AB - STUDY OBJECTIVE: We assess accuracy of ECG interpretation and indications for obtaining ECGs and develop a clinical classification system of ECG abnormalities. METHODS: Prospectively acquired ECG data on patients 0 to 21 years of age and presenting to our pediatric emergency department (ED) were obtained. Clinical indications were documented. The initial ECG interpretation (pediatric ED attending physician) was compared with the criterion standard (pediatric cardiologist). A blinded cardiology panel reviewed discrepancies, and a final concordance rate was determined. An ECG abnormality classification system was developed and used to categorize these abnormal ECGs. RESULTS: One thousand six hundred fifty-three ECGs from 1,501 patients, aged 2 days to 21 years (median 10.0 years), were obtained during 3.5 years. Fifty-one percent were male patients. ECG indications included chest pain (21%), seizure or syncope (18%), arrhythmias (17%), apparent life-threatening event or respiratory symptoms (16%), ingestions (10%), cardiac abnormality (10%), and miscellaneous (8%). From 1,631 ECGs, 1,160 (71%) were normal (class 0), 259 (16%) were minimally abnormal (class I), 174 (11%) were moderately abnormal (class II), and 38 (2%) were severely abnormal (class III). Kendall's tau-b test showed concordance of 0.73 (95% confidence interval 0.70 to 0.77) between pediatric ED and cardiology interpretation. The sensitivity of pediatric ED interpretation was 75%, and the specificity was 98.5%. The positive predictive value of pediatric ED interpretation was 88.3%, and the negative predictive value was 96.3%. CONCLUSION: We conclude that, overall, a high rate of concordance exists between the pediatric emergency physician's and the cardiologist's ECG interpretation. The majority of discordant ECGs are not clinically significant. However, among the clinically significant ECGs, there is a higher rate of discordance. These data suggest that review of pediatric ECGs by pediatric cardiologists may significantly reduce underdetection of clinically important ECG findings in children. PMID- 16308066 TI - Out-of-hospital pediatric cardiac arrest: an epidemiologic review and assessment of current knowledge. AB - STUDY OBJECTIVE: We systematically summarize pediatric out-of-hospital cardiac arrest epidemiology and assess knowledge of effects of specific out-of-hospital interventions. METHODS: We conducted a comprehensive review of published articles from 1966 to 2004, available through MEDLINE, Cumulative Index to Nursing and Allied Health Literature, EmBase, and the Cochrane Registry, describing outcomes of children younger than 18 years with an out-of-hospital cardiac arrest. Patient characteristics, process of care, and outcomes were compared using pediatric Utstein outcome report guidelines. Effects of out-of-hospital care processes on survival outcomes were summarized. RESULTS: Forty-one studies met inclusion criteria; 8 complied with Utstein reporting guidelines. Included in the review were 5,363 patients: 12.1% survived to hospital discharge, and 4% survived neurologically intact. Trauma patients (n=2,299) had greater overall survival (21.9%, 6.8% intact); a separate examination of studies with more rigorous cardiac arrest definition showed poorer survival (1.1% overall, 0.3% neurologically intact). Submersion injury-associated arrests (n=442) had greater overall survival (22.7%, 6% intact). Pooled data analysis of bystander cardiopulmonary resuscitation and witnessed arrest status showed increased likelihood of survival (relative risk 1.99, 95% confidence interval 1.54 to 2.57) for witnessed arrests. The effect of bystander cardiopulmonary resuscitation is difficult to determine because of study heterogeneity. CONCLUSION: Outcomes from out-of-hospital pediatric cardiac arrest are generally poor. Variability may exist in survival by patient subgroups, but differences are hard to accurately characterize. Conformity with Utstein guidelines for reporting and research design is incomplete. Witnessed arrest status remains associated with improved survival. The need for prospective controlled trials remains a high priority. PMID- 16308067 TI - Out-of-hospital pediatric cardiac arrest: where are we now and where do we need to go? PMID- 16308068 TI - Emergency physicians' fear of malpractice in evaluating patients with possible acute cardiac ischemia. AB - STUDY OBJECTIVE: We evaluate the association between emergency physicians' fear of malpractice and the triage and evaluation patterns of patients with symptoms suggestive of acute coronary syndrome. METHODS: We surveyed 33 emergency physicians of 2 university hospitals during the preintervention phase of an implementation trial of the Agency for Health Care Policy and Research Unstable Angina guideline in 1,134 study patients. The survey included a 6-item instrument that addressed concerns about malpractice and a measure of general risk aversion. We used hierarchical logistic regression to model emergency department (ED) triage decisions and diagnostic testing as a function of fear of malpractice, with adjustment for patient characteristics, Agency for Health Care Policy and Research guideline risk group, study site, and clustering by emergency physician. RESULTS: Overall, emergency physicians in the upper tertile of malpractice fear were less likely to discharge low-risk patients compared with emergency physicians in the lower tertile (adjusted odds ratio [OR] 0.34; 95% confidence interval [CI] 0.12 to 0.99; P=.05). Patients treated by emergency physicians in this group were also more likely to be admitted to an ICU or telemetry bed (adjusted OR 1.7; 95% CI 1.2 to 2.4). In addition, emergency physicians in the upper tertile of malpractice fear were more likely to order chest radiography, as well as cardiac troponin. Malpractice fear accounted for a similar amount of variance after controlling for emergency physicians' risk aversion. CONCLUSION: Malpractice fear accounts for significant variability in ED decisionmaking and is associated with increased hospitalization of low-risk patients and increased use of diagnostic tests. PMID- 16308069 TI - Decisions, decisions: emergency physician evaluation of low probability-high morbidity conditions. PMID- 16308070 TI - What do health savings accounts mean for the emergency department? AB - The insurance market is evolving, with increased emphasis on plans with high deductibles and a large degree of coinsurance. This article serves as an introduction to the defining characteristics of consumer-driven health care plans and their associated health savings accounts. We discuss the most recent evidence on the adoption of these plans and their effects on use and reimbursement. Compared to many specialties, the emergency department (ED) may be insulated from extensive shopping and price negotiation, because visits to the ED are often for urgent and time-sensitive conditions. However, ED utilization patterns may change if cost-conscious health savings account holders forgo other necessary medical care, or if they seek out substitutes to the ED for less urgent problems. In the long run, the ED may feel the impact of changes that stem from 2 areas: the ability of health savings accounts to control the increase in health care costs, and the potential of health savings accounts to replace or undermine more comprehensive health insurance plans. We note areas that emergency physicians should monitor as health savings accounts become more prominent. PMID- 16308071 TI - Once upon a time in the emergency department: a cautionary tale. PMID- 16308072 TI - Epidemiologic patterns of injuries treated in ambulatory care settings. AB - STUDY OBJECTIVE: Epidemiologic studies of injury morbidity have relied primarily on data from emergency departments (EDs) and hospital admissions. We seek to assess the incidences and characteristics of acute injuries treated at EDs and other ambulatory care settings. METHODS: Using data from the 2002 National Health Care Survey on initial visits for acute injuries to EDs, physician offices, and hospital outpatient departments, we estimated the frequencies and incidence rates of medically attended injury by patient characteristics and care setting. RESULTS: In the United States in 2002, 76 million nonfatal acute injuries received initial medical attention at EDs (46.2%), physician offices (47.8%), and outpatient departments (6.0%). The overall annual incidence rate of medically attended injury was 26.8 per 100 population (95% confidence interval 24.4 to 29.7). Falls accounted for 16.7% of all medically attended injuries. Injury patients who were black or uninsured were significantly more likely to visit EDs than other care settings for treatment. More than 2.4 million (3.2%) injury patients were admitted to hospitals, 96.6% of them through EDs. CONCLUSION: Fewer than half of all medically attended acute injuries in the United States receive initial treatment in EDs. Injury severity and characteristics vary among care settings. PMID- 16308073 TI - National Highway Traffic Safety Administration (NHTSA) notes. Breath test refusals in DWI enforcement: an interim report. PMID- 16308074 TI - The advocate. PMID- 16308075 TI - Images in emergency medicine. Acute left-sided rectus sheath hematoma. PMID- 16308076 TI - Images in emergency medicine. Crohn's disease. PMID- 16308077 TI - Anthrax scare 2005, Washington, DC: operational issues in chemoprophylaxis. PMID- 16308078 TI - Serum lactate as a predictor of mortality in emergency department patients with infection: does the lactate level tell the whole story? PMID- 16308082 TI - An antifungal peptide from the coconut. AB - A chromatographic procedure consisting of ion exchange chromatography on DEAE cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on CM-cellulose, and gel filtration by fast performance liquid chromatography on Supedex 75 was utilized to isolate a 10 kDa antifungal peptide from coconut flesh. The peptide was unadsorbed on DEAE-cellulose, but adsorbed on Affi-gel blue gel and CM-cellulose. It displayed antifungal activity against Fusarium oxysporum, Mycosphaerella arachidicola and Physalospora piricola. The IC50 values of its inhibitory activities on mycelial growth in M. arachidicola and HIV-1 reverse transcriptase activity were respectively 1.2 and 52.5 microM. PMID- 16308084 TI - Breast fine needle aspiration biopsy: prevailing recommendations and contemporary practices. AB - In 1996, a National Cancer Institute conference was held in Bethesda,Maryland to define parameters for the practice of breast fine needle aspiration (BFNA). Representatives of the American Society of Cytopathology, Papanicolaou Society of Cytopathology, American College of Radiology, American College of Obstetricians & Gynecologists, Society of Surgical Oncology, American Academy of Family Physicians, College of American Pathologists, National Consortium of Breast Centers, International Academy of Cytology, American Society of Clinical Pathologists, American Cancer Society, American College of Surgeons, and American Society for Cytotechnology developed and reviewed recommendations. These guidelines were referred to as "The Uniform Approach to Breast Fine Needle Aspiration Biopsy." This article reviews these recommendations and the contemporary evolution of the practice of BFNA since their original publication. PMID- 16308085 TI - Image-directed fine needle aspiration biopsy in nonpalpable breast lesions. AB - Fine needle aspiration biopsy of nonpalpable breast lesions may be performed with imaging guidance. The technique has a potential role, but is underused because of a relative paucity of specialty-trained, enthusiastic breast cytopathologists. This article discusses the strengths and limitations of the technique in the context of the various imaging modalities, and in comparison with needles used for histologic sampling. PMID- 16308086 TI - Core needle biopsy versus fine needle aspiration biopsy: are there similar sampling and diagnostic issues? AB - Evidence suggests that in experienced hands fine needle aspiration biopsy (FNAB) is highly safe and effective for the evaluation of patients who have palpable breast lesions. In most cases, FNAB leads to an appropriate clinical or surgical management. FNAB can also be effectively used in evaluation of cystic lesions under ultrasound guidance. Other nonpalpable lesions can benefit from image directed core needle biopsy (CNB). Overall, cost should influence the decision to use FNAB or CNB. FNAB may be the only affordable procedure in developing countries. Breast cancer affects significant numbers of women worldwide. Attempts should be made to take all the measures necessary to consider optimal approaches to breast health care for everyone regardless of age, race, ethnicity, or social status. PMID- 16308087 TI - Cytomorphology of benign breast disease. AB - Fine needle aspiration biopsy is a widely used technique for the initial diagnosis of mammary lesions. The majority of patients undergoing fine needle aspiration biopsy of a breast lesion will have a benign disease of the breast. This article provides a review of the cytomorphologic features in a variety of benign breast lesions,and discusses the commonly encountered differential diagnoses on aspiration biopsy. Topics discussed include inflammatory and reactive conditions, treatment-induced changes, pregnancy-related changes, benign proliferative lesions, and benign neoplasms of the breast. PMID- 16308088 TI - Cytomorphology of fibrocystic change, high-risk proliferative breast disease, and premalignant breast lesions. AB - In a prospective study using mammographically guided fine needle aspirates in 100 nonpalpable breast lesions, the author's group assessed the reliability of a cytological grading system to define the cytological features of proliferative and nonproliferative breast disease and to differentiate between benign, premalignant and malignant breast lesions. We developed a cytological grading system evaluating the aspirates for the cellular arrangement, the degrees of cellular pleomorphism and anisonucleosis, presence of myoepithelial cells and nucleoli and the status of the chromatin pattern. This grading system, now recognized as the Masood Cytology Index, is commonly used as a surrogate end point biomarker in chemoprevention trials. PMID- 16308089 TI - Cytomorphology of common malignant tumors of the breast. AB - A definitive cytologic diagnosis of breast cancer is usually possible when using the six major criteria of malignancy (cellularity, dyshesion, monomorphism, anisonucleosis, irregular nuclear membranes, prominent nucleoli) as part of the triple test. Carcinomas of special type have unique clinical and cytologic features that pathologists need to consider, because these may confuse interpretation. Complete subtyping of carcinomas may not always be possible by fine needle aspiration. Diagnostic accuracy for breast carcinoma is excellent. False-negative diagnoses are infrequent and chiefly due to sampling issues. False positive diagnoses are extremely rare. Uniform report terminology should be used to ensure that diagnostic information is conveyed appropriately and consistently to guide the next diagnostic or treatment step. PMID- 16308090 TI - Cytomorphology of rare malignant tumors of the breast. AB - This article reviews cytomorphology of rare malignant tumors of the breast: squamous carcinoma, metaplastic carcinoma, adenoid cystic carcinoma, apocrine carcinoma, secretory carcinoma, lipid-rich carcinoma, and carcinoma with choriocarcinomatous features. It is important to bear in mind the cytomorphology and diagnosis of rare malignant tumors of the breast in analysis of breast fine needle aspiration smears. Although rare, these malignant tumors can be diagnosed by the cytomorphological characteristics, and should be included in the differential diagnoses of breast neoplasms. PMID- 16308091 TI - Medical-legal issues associated with breast fine needle aspiration biopsy. AB - This article discusses medical and legal issues associated with breast fine needle aspiration biopsy and develops risk-management considerations for clinical practice. Basic legal considerations are first discussed, including the law of negligence as it applies to breast fine needle aspiration. The variable concept of duty to patients is reviewed. Communication of diagnosis, both formally and informally, in the medical setting is covered. The claims review process is then discussed. The meaning of misdiagnosis, including overdiagnosis and delay of diagnosis, is discussed in terms of legal harm and expert review. PMID- 16308092 TI - Nipple fluid cytology. AB - With increasing emphasis on public awareness of breast cancer and screening mammography, more women seek consultation for breast symptoms, including nipple discharge. The presence of nipple discharge is distressing for the patient; however, its origin is fortunately most often benign. Nipple discharge associated with a recognizable mass requires sampling and surgical excision. Currently, clinical breast examination, cytology, ductography, mammogram, and ultrasound are commonly used to evaluate patients who have nipple discharge. Ductal lavage and ductoscopy, in association with emerging new technology, may be of help in increasing the sensitivity of nipple fluid cytology. In addition, nipple fluid cytology may offer a new way to study the spectrum of premalignant breast lesions, and to identify women at risk for breast cancer. PMID- 16308093 TI - Intraoperative imprint cytology in assessment of sentinel lymph nodes and lumpectomy surgical margins. AB - Intraoperative imprint cytology (IIC) in the assessment of sentinel lymph nodes (SNs) allows immediate, cost-effective axillary lymph node dissection. IIC diagnosis is accurate in up to 100% of grossly abnormal SNs. Despite overall low sensitivity for grossly negative SNs, the benefits of immediate complete axillary lymph node dissection offset the increased risk of missing micrometastases or loss of isolated tumor cells (ITCs) by performing frozen section. IIC of the lumpectomy margins is rapid, accurate, and cost effective. It allows re-excision during initial surgery if needed with better cosmetic result. It is a useful adjunct to, and frequently a replacement for frozen section in many centers. Cytopathologists must familiarize themselves with both advantages and pitfalls of IIC to avoid errors. PMID- 16308094 TI - Prognostic/predictive factors in breast cancer. AB - This can be an exciting time for pathologists and cytopathologists as we refine or knowledge of prognostic/predictive factors in breast cancer. We can become more visible in our role as consultants to the other physicians, and more engaged in our role as re-searchers. Recent advances in computer science, coupled with the availability of new biological markers, now provide unique opportunities for us to expand our diagnostic abilities and also predict the biologic behavior of a given tumor. Thus, we must become more familiar with emerging concepts and technologies in different disciplines. PMID- 16308095 TI - Cytomorphology as a risk predictor: experience with fine needle aspiration biopsy, nipple fluid aspiration, and ductal lavage. AB - Primary prevention of breast cancer requires identification and elimination of cancer-causing agents, which is an incredibly difficult task to follow. Secondary prevention involves screening individuals who are at increased risk for breast cancer in hopes that early intervention will affect survival. In the 1980s, chemoprevention received serious attention. This approach was aimed at reducing cancer risk by administration of natural or synthetic clinical compounds that prevent, reverse, or suppress carcinogenesis in individuals at increased risk for cancer. It was not until 1998, however, when the first report from the National Surgical Adjunct Breast and Bowel Project (Breast Cancer Prevention Trial BCPT; P 1) randomized clinical trial appeared in the literature supporting the hypothesis that breast can-cer can be prevented. This study showed that administration of tamoxifen reduced the risk for invasive and noninvasive breast cancer by almost 50% in all age groups. With the current availability of tamoxifen as a chemopreventive agent and with the increasing emphasis on early breast cancer detection and prevention, more women seek consultation to determine their risk for breast cancer. However, in the absence of any detectable breast lesion, clinically and mammographically, only a few women may volunteer to have their breasts sampled by surgical biopsy for risk assessment. Other non-surgical procedures include fine needle aspiration biopsy (FNAB), nipple aspirate fluid (NAF), and the recently introduced procedure, ductal lavage. These techniques may provide better alternatives. These minimally invasive procedures are capable of recruiting cellular material for cytomorphologic interpretation and biomarker studies. PMID- 16308096 TI - The future prospect: ductoscopy-directed brushing and biopsy. AB - The intraductal approach to breast cancer and premalignant lesions has now developed to yield substantial cytologic samples of exfoliated cells. Standard cytology is still inadequate in sensitivity and specificity to accurately interpret the majority of samples. As techniques evolve using ductoscopic biopsy and molecular marker panels to increase accuracy of cytologic interpretation, these tools will be able to unravel the breast carcinogenesis pathways. They will also offer considerable benefit in screening for premalignant changes and developing effective chemoprevention strategies. PMID- 16308097 TI - Concordance between qualitative and quantitative cultures in burned patients. Analysis of 2886 cultures. AB - OBJECTIVE: To determine the concordance between superficial cultures (SC) and quantitative cultures (QC) in the diagnosis of wound infection in burn patients. METHOD: SAMPLE: All SC and QC taken from the same patient, site and during the same surgery were analysed. VARIABLES: On the SC, the microorganism (MO) and its amount defined subjectively by the microbiologist was recorded (negative, very low, low, regular and abundant). On the QC, the MO and its amount were expressed as colony forming units per gram of tissue (CFUs/g). STATISTICS: Kappa index of agreement beyond chance; Wilcoxon and Kruskall-Wallis for continuous variables and chi(2) for categorical variables were used with a p<0.05 indicating statistical significance. RESULTS: One thousand four hundred and forty three pairs of cultures were analyzed. The concordance between SC and QC (Kappa index) was 52%. On the SC, only when the microbiologist subjectively informed "abundant" MOs there was a significant difference (p<0.0001). There were 6.1% of QCs with more than 10(5) CFUs/g and the most frequent MOs isolated were: S. aureus (27.9%), E. coli (11.6%), P. aeruginosa (11.6%), E. faecalis (11.6%) and S. epidermidis (7.0%). CONCLUSIONS: SC has a moderate concordance with the QC showing a low reliability between the two methods. The subjective information given by the microbiology technician in the SC is not precise. A study in which the two methods be compared blindly against the reference standard, in a prospective cohort of patients, it is needed to discriminate which of two methods it is the most accurate one determining sensitivity and specificity. PMID- 16308098 TI - Burn first aid in Western Australia--do healthcare workers have the knowledge? AB - Burn injuries treated with adequate immediate first aid are associated with more favourable outcomes, limiting tissue damage and subsequent morbidity including the need for surgery. Cool running water at a temperature of between 10-15 degrees C for 20 to 30 minutes is considered adequate burn first aid treatment. A prospective audit of all new patients (n=227) with burns, attending the minor burn facility at Royal Perth Hospital showed only 88 (39%) patients received appropriate first aid. Fifty percent of patients receiving inappropriate first aid, had this delivered by his or her primary health care contact. This study aims to determine the knowledge of burns first aid among healthcare workers (HCW) and compare this to the general population. A customized survey was performed, four sample cases were included with 4 possible answers in a multiple choice format. All case studies asked the participant to record immediate first aid management. On a case by case basis, burn first aid knowledge was fair but overall knowledge very poor, only 18.8% of respondents achieving 4 correct responses. The uptake of first aid courses was high among HCW at 75% but particularly low among the NHCW at 28%. Our study has shown the value of performing such a course, with a statistically significant (p=0.00) difference between participants who had completed a first aid course and those that had not. The purpose of this study was to quantify knowledge of burns first aid with a view to providing directed education, studies have shown the success of multimedia campaigns. It has been shown that good first aid improves the outcome for the burn patient and this study demonstrates that participation in a first aid course improves knowledge. It is justifiable to progress toward compulsory first aid courses which include first aid of the burns patient to improve outcomes for the community as a whole. PMID- 16308099 TI - Lightning injuries in a pregnant woman: a case report and review of the literature. PMID- 16308100 TI - The use of topical negative pressure in a paediatric patient with extensive burns. PMID- 16308101 TI - SELDI-MS-based expression profiling of ductal invasive and lobular invasive human breast carcinomas. AB - Expression profiling using proteomic techniques has a great potential to identify new biomarkers that might help to better diagnose and treat diseases such as breast cancer, which is one of the leading causes of cancer death in women. Surface-enhanced laser desorption ionization mass spectrometry (SELDI-MS) combines chromatographic separation of peptides and proteins with mass spectrometry and is a fast, user-friendly tool to analyze protein and peptide profiles. SELDI-MS was employed for a comparative analysis of lobular invasive versus ductal invasive breast tumors to find differentially expressed proteins and peptides, and to validate this technique for biomarker identification using complex samples such as tissue. After optimization of sample preparation using HMEC and MCF-7 cell lines, 20 breast tumors were analyzed, and about 550 mass signals corresponding to an estimated 140 native peptides and proteins were detected in each tumor. Only 14% of the mass signals were present in more than six tumors of one subgroup or in more than 12 tumors of both groups showing a great overall heterogeneity of the peptide and protein profiles obtained. Peptide mass signals specific for each of the analyzed groups were identified. In addition, we detected peptides from laser-microdissected ductal invasive and intraductal tumor parts corresponding to peptides present in whole tumors. The low amount of identified peptides and proteins and the observed heterogeneity suggest that SELDI-MS is not well suited for biomarker identification of and profiling experiments on complex samples such as tumor tissue. PMID- 16308102 TI - Differential Prox-1 and CD 31 expression in mucousae, cutaneous and soft tissue vascular lesions and tumors. AB - The study of lymphatic vessels and lymphatic tumors has been hampered with difficulty due to the overlapping morphological features between blood and lymphatic endothelial cells, as well as to the lack of specific lymphatic endothelial markers. Over the last few years, lymphatic vessels and lymphangiogenesis have received great attention owing to their putative implications in terms of metastatic dissemination and the promise of targets for lymphangiogenic therapy. Prox-1 is a nuclear transcription factor that plays a major role during embryonic lymphangiogenesis and is deemed to be a useful marker for differentiating lymphatic endothelial cells from the other blood vessels endothelial cells. Here, we describe a double-immunostaining strategy for formalin-fixed, paraffin-embedded tissues that aims at evaluating the distribution of Prox-1 and CD 31 - a cytoplasmic pan-endothelial marker - in a series of 28 mucousae, cutaneous and soft tissue vascular lesions and tumors, including hemangiomas, lymphangiomas, lymphangiectasia, and Kaposi's sarcomas. Our results showed that in non-lesional mucousae and skin, Prox-1 decorated exclusively the nuclei of endothelial cells in lymphatic vessels. Prox-1 stained almost all the benign lymphatic vascular lesions/tumors (91%) and was absent or only focally positive in 75% of blood vascular tumors. CD 31 stained endothelial cells of blood vessels of superficial and deep dermal plexuses, lymphatics, and all blood vascular lesions/tumors. Kaposi's sarcomas were all positive for both CD 31 and Prox-1 markers. In conclusion, although Prox-1 expression in vascular lesions/tumors was not entirely restricted to tumors with known lymphatic differentiation, CD 31/Prox-1 double-immunolabeling can be used as an adjunct marker to identify lymphatic vessels in routinely processed formalin-fixed, paraffin-embedded samples. PMID- 16308103 TI - Differential expression of Janus kinase 3 (JAK3), matrix metalloproteinase 13 (MMP13), heat shock protein 60 (HSP60), and mouse double minute 2 (MDM2) in human colorectal cancer progression using human cancer cDNA microarrays. AB - In this study, we applied commercially available cDNA microarray systems (1068 genes) to investigate the genetic changes in six colorectal cancers (CRC). Thirty two genes fell into the group of commonly upregulated genes. In addition, we immunohistochemically investigated the expression of the four top ranked upregulated genes, Janus kinase 3 (JAK3), matrix metalloproteinase 13 (MMP13), heat shock protein 60 (HSP60), and mouse double minute 2 (MDM2), in 44 CRC. JAK3 staining was located in the cancer cells. A comparison of JAK3 immunostaining and clinicopathological parameters showed a significant association of tumor differentiation, pT, and TMN stage. Staining of MMP13 and HSP60 was noted mainly in the cytoplasm of cancer cells. A significant association of these expressions was observed with tumor differentiation and pT. MDM2 staining was noted in the nucleus of cancer and non-cancer cells. No significant association of clinicopathological parameters with MDM2 expression was observed. In multivariate analysis, JAK3 immunoreactivity showed independent prognostically unfavorable predictors. These data suggest that JAK3, in particular, is a highly significant, prognostic immunohistochemical marker in CRC. This study proves that cDNA microarrays, plotted by a small number of genes from a few samples, are both practical and useful. PMID- 16308104 TI - Localized fibrous tumors (LFTs) of the pleura: clinical data, asbestos burden, and syntactic structure analysis applied to newly defined angiogenic/growth regulatory effectors. AB - This study was performed to add clinical data, to introduce new markers, and to perform syntactic structural analysis on localized fibrous tumors (LFTs) of the pleura. The material comprised clinical data and processed sections obtained from 36 patients. The results achieved from quantitative imaging techniques and syntactic structure analysis were correlated with clinical data, including patients' habits (smoking), asbestos exposure, survival, and tumor recurrence. The disease caused increasing chest pain and dyspnea in 47% of patients. Exposure to asbestos was noted in 13 out of 36 patients, whereas smoking posed no major risk factor. Two patients developed a recurrent tumor after 8 and 42 months, respectively; none of the other patients died of this tumor disease within the follow-up period of maximal 212 months. The cases were clearly discriminated from mesotheliomas by the marker profile. Frequent expression of accessible ligands for endogenous lectins galectins-1 and -3, the expression of the angiogenic macrophage migration inhibitory factor (MIF), and the dense vascularization intimate a functional relationship. The proliferation index (Nv) was computed to be 1.6% in line with the balance of galectin expression. Abnormal p53 was expressed in only 19.4% of the cases. The diagnosis of LFT can be aided by quantitative assessment of vimentin, CD34, MIF, vascularization, and proliferation. Considering the galectin network, differential expression was noted with preference to effectors limiting growth and aggressiveness. PMID- 16308105 TI - Expression of CD3 antigens in renal tubule epithelium and renal oncocytomas. AB - CD3 antigen, formerly thought to be specific for T lymphocytes, has been identified in Purkinje cells of the cerebellum and gastric parietal cells in several species, including humans. The antibodies commonly used to recognize CD3 are directed against the epsilon-subunit of the T cell receptor. This subunit has a role in signal transduction in T lymphocytes and possibly other types of cells. We immunostained sections for CD3 from normal kidneys of several species, including humans, and from different primary human renal cortical neoplasms to determine if CD3 antigen is expressed in normal and in neoplastic tubular epithelium. CD3 expression was strong in normal proximal and distal tubular epithelium in most species and in renal oncocytomas, weak in chromophobe carcinoma, and negative in clear cell carcinomas, in papillary renal cell carcinoma, and in a transitional cell carcinoma. These findings suggest that this marker may be useful in the diagnostic workup and classification of renal cortical neoplasms. PMID- 16308106 TI - Roles of MMP-2/-9 in cardiac dysfunction during early multiple organ failure in an ovine animal model. AB - Biventricular dilation and severe cardiac dysfunction are observed during septic shock. However, when endotoxemia and vasoconstrictor-masked hypovolemia work in concert in the pathogenesis of shock, the clinical scenario is more adverse compared to one of the insults acting alone. Matrix metalloproteinases (MMPs) are involved in chronic and acute heart failure by degrading the mechanical scaffold of the heart and several intracellular proteins. Therefore, the roles of MMP-2, MMP-9, MT1-MMP, focal adhesion kinase (FAK), and Paxillin in hearts of early multiple organ failure induced by norfenefrine-masked hypovolemia and endotoxemia were investigated in an ovine model. Experimental groups included (1) norfenefrine-masked hypovolemia plus endotoxemia (NMH+ENDO) (n=6), (2) norfenefrine-masked hypovolemia without endotoxemia (NMH) (n=6), (3) recurrent endotoxemia during normovolemia (ENDO) (n=6), and (4) healthy untreated controls (CON) (n=3). Apoptosis was determined by TUNEL-staining. Gel zymography revealed significantly increased MMP-2 activity in NMH+ENDO compared to ENDO and controls. MMP-9 activity was significantly elevated in all experimental groups. MMP-2 was significantly increased at the protein level, while MMP-9 was unaltered. MT1-MMP was not significantly changed in any group. Increased MMP activities were associated with cardiac deterioration. MMP-2/-9 activity and phosphorylated Paxillin (p-Paxillin) expression correlated positively with cardiomyocyte apoptosis. This study underscores the pivotal roles of MMP in acute cardiac dysfunction during early multiple organ failure in combined vasoconstrictor masked hypovolemic and endotoxemia shock. PMID- 16308107 TI - Fallopian tube prolapse misdiagnosed as vault granulation tissue: a report of three cases. AB - Prolapse of the fallopian tube into the vagina is an uncommon complication caused by either vaginal or abdominal hysterectomy. Recently, however, we encountered three cases with prolapse of the fallopian tube after abdominal hysterectomy. The patients presented with vaginal bleeding. A red hemorrhagic granular mass, misdiagnosed as vaginal granulation tissue both macroscopically and microscopically, was noted at the apex of vagina. Pathologically, one case was initially diagnosed as vaginal vault granulation tissue, but there were two recurrences after excision. Microscopically, the mass had a papillary or villous outer surface with a complex pattern of tubular and glandular structures, as well as acute and chronic inflammatory infiltrates in the fibrovascular stroma. A typical ciliated tubal type of epithelium was identified, and on immunohistochemical staining for cytokeratin, attenuated epithelial cells were detected. It is necessary to receive a pathologic confirmation by performing vaginal biopsy when fallopian tube prolapse is clinically suspected, thus preventing misdiagnosis. PMID- 16308110 TI - Bradydysrhythmias and atrioventricular conduction blocks. AB - Bradydysrhythimas include sinus bradycardia, junctional brady-cardia, and idioventricular rhythm, which can be distinguished by examining the tracing for the presence or absence of P waves,noting the morphology of these P waves, and determining the width of the QRS complex. Sinoatrial blocks may occur in either first, second, or third degree varieties. Only second degree sinoatrial block can be detected on the 12-lead ECG. Sinus pause and sinus arrest may mimic second degree sinoatrial block, but their periodicity is irregular. The cyclic variability of sinus arrhythmia is unique; as with the other bradydysrhythmias, it may be innocent or pathologic depending upon clinical circumstances. Atrioventricular blocks may occur, and, similar to sinoatrial blocks, they are also categorized as first-, second-, or third degree. These are of greater clinical relevance than their sinoatrial counterparts. PMID- 16308111 TI - Tachydysrhythmias. AB - Tachydysrhythmias arise from different mechanisms that can be characterized as being caused by re-entrant circuits, enhanced or abnormal automaticity, or triggered after-depolarizations. The approach to the tachydysrhythmia should begin with distinguishing sinus from non-sinus rhythms, then assessing QRS complex width and regularity. This article review tachydysrhythmias. PMID- 16308112 TI - Intraventricular conduction abnormalities. AB - There are multiple types of intraventricular conduction abnormalities, each with its own unique clinical significance. It is useful to categorize conduction abnormalities, or blocks, by the number of fascicles involved. This article reviews intraventricular conduction abnormalities. PMID- 16308113 TI - Acute coronary syndromes. AB - Despite technologic advances in many diagnostic fields, the 12-lead ECG remains the basis for early identification and management of an acute coronary syndrome. This article reviews the use of the ECG in acute coronary syndromes. PMID- 16308114 TI - ST segment and T wave abnormalities not caused by acute coronary syndromes. AB - This article reviews the ST segment and T wave abnormalities seen in non-acute coronary syndrome (ACS) electrocardiograph presentations. Particular emphasis is placed on the distinction of these non-ACS syndromes from acute coronary syndrome related ST segment and or T wave change. PMID- 16308115 TI - ECG manifestations: noncoronary heart disease. AB - The electrocardiogram reflects changes to the heart beyond those seen in acute myocardial infarction and acute coronary syndromes. Diseases of the pericardium and heart muscle such as pericarditis,myocarditis, and pericardial effusion have characteristic manifestations. Hypertensive heart disease is associated with a variety of changes on the electrocardiogram, as is valvular heart disease. Cardiac rhythm disturbances have been associated with the Brugada syndrome and the long QT syndrome, both of which have telltale findings on the electrocardiogram. The manifestations of dextrocardia, although rare, should be familiar to those who interpret electro-cardiograms. Transplanted hearts also feature classic changes, both in health and in stages of rejection. The various electrocardiographic manifestations of these noncoronary heart diseases are reviewed here. PMID- 16308116 TI - ECG manifestations of selected extracardiac diseases. AB - This article reviews the ECG manifestations of selected extracardiac diseases, including pulmonary embolism, pneumothorax, pulmonary hypertension, aortic dissection, central nervous system dis-ease, gastrointestinal disease, and sarcoidosis. PMID- 16308117 TI - ECG manifestations of selected metabolic and endocrine disorders. AB - Alterations in serum calcium and potassium concentration may manifest as detectable changes on the ECG and alert the emergency physician to the presence of an underlying electrolyte disturbance. In addition, various endocrine abnormalities, metabolic disturbances, and environmental emergencies may cause alterations in the ECG. PMID- 16308118 TI - ECG manifestations: the poisoned patient. AB - Despite that drugs have widely varying indications for therapeutic use, many unrelated drugs share a common cardiac pharmacologic effect if taken in overdose. The purpose of this article is to group together agents that cause similar electrocardiographic effects,review their pharmacologic actions, and discuss the electrocardiographic findings reported in the medical literature. PMID- 16308119 TI - Electronic pacemakers. AB - The number of patients who have cardiac pacemakers has increased markedly over the past few decades since the technology was first introduced in the 1950s. Advances in technology, expanding indications, and the aging of the population ensure that clinicians will encounter patients with cardiac pacemakers on a regular basis. This article summarizes the electrocardiographic manifestations of the normally functioning permanent cardiac pacemaker and abnormalities associated with pacemaker malfunction. PMID- 16308120 TI - The pediatric ECG. AB - Knowledge of the basics of pediatric ECG interpretation is helpful in differentiating normal from abnormal findings. These basics include familiarity with the age-related normal findings in heart rate, intervals, axis, and waveform morphologies; an understand-ing of cardiac physiologic changes associated with age and maturation, particularly the adaptation from right to left ventricular predominance; and a rudimentary understanding of common pediatric dysrhythmias and findings associated with congenital heart diseases. PMID- 16308121 TI - ECG techniques and technologies. AB - The electrocardiogram (ECG) continues to be a critical component of the evaluation of patients who have signs and symptoms of emergency cardiac conditions. This tool is now approximately 100 years old and has been a standard in clinical practice for more than half a century. Application of new signal processing techniques and an expansion in the use of additional leads allows clinicians to extract more and more information from the cardiac electrical activity. An understanding of the technology inherent in the recording of ECGs allows one to more fully understand the benefits and limitation of electrocardiography. PMID- 16308122 TI - Electrode misconnection, misplacement, and artifact. AB - This article includes a discussion of limb electrode misconnection,precordial electrode misconnection and misplacement, and electro-cardiographic artifact. PMID- 16308123 TI - What have you survived? PMID- 16308124 TI - Exploring parental grief: combining quantitative and qualitative measures. AB - PURPOSE: This study explores parental grief on the death of a child of any age. STUDY DESIGN: The sampling frame for the study consisted of 74 respondents reporting that they experienced the death of a living child. With the exception of standard demographic measures, the quantitative and qualitative items in the instrument were designed specifically for this project. The content validity of the instrument was assessed by a panel of experts on grief. Key quantitative items related to child loss and grief were formatted with between three- and five point ordinal answer categories. Two types of qualitative items were developed by the authors. The reliability of the instrument designed for this study was not determined. RESULTS: This survey of parents who experienced the death of a child during their lifetime explores and extends current understanding of the complex emotional response of grief. The study offers empirical support for the notion of grief as ongoing in the life of a parent whose child had died. The findings have significant implications for further clinical research supporting studies to explore commonalities in the experience of grieving families regardless of the cause of and time since the death of their child. In addition, the findings may serve to inform the development and provision of services for bereaved parents. PMID- 16308125 TI - Treatment of depressive disorders in split versus integrated therapy and comparisons of prescriptive practices of psychiatrists and advanced practice registered nurses. AB - This study examined the differences in adherence to medication for the treatment of depressive mood disorders whether the patient was involved in split or integrated therapy and if the patient was being treated by a psychiatrist or an advanced practice registered nurse (APRNs) in psychiatry with prescriptive authority. The sample consisted of 122 adults ages 20-60 who carried a diagnosis of major depression, dysthymia, or bipolar II disorder, voluntarily treated in the private sector. Chi-square was the measure used to assess differences in adherence. Adherence was determined by documentation from chart reviews, retrospectively, of the prescriptions for psychotropic medication for a period of up to nine months for each patient included in the research. Six psychiatrists and six APRNs volunteered to collect data from their private practices. Findings demonstrated no statistically significant differences in patient's adherence to medication if they were in split or integrated treatment or if either psychiatrists or an APRN treated them. Psychiatrists used more of secondary class of antidepressants and more antianxiety agents than did the APRNs. The nurses spent more time with patients, using more integrated therapy, than did the psychiatrists. PMID- 16308127 TI - The transition from pregnancy to postpartum in previously infertile women: a focus on depression. AB - Women with previous infertility may be at risk for developing depression during pregnancy (Olshansky, E. (2003). A theoretical explanation for previously infertile mothers' vulnerability to depression. Journal of Nursing Scholarship, 35(3), 23-268) and postpartum, a possibility often overlooked in this population. There is a dearth of research about depression in women during the transition from pregnancy to postpartum. This pilot longitudinal, cohort design study examined this transition, focusing on potential predictors of postpartum depression (PPD) in previously infertile women. Two significant variables, marital satisfaction and the "divided self," a condition in which women present an outer compliant self while experiencing internal anger, may be predictors of the development of PPD in new mothers with a history of infertility. Additional research is needed to confirm these preliminary findings. PMID- 16308126 TI - Design and evaluation of an online teaching strategy in an undergraduate psychiatric nursing course. AB - Psychiatric nurse educators are challenged to prepare graduates in meeting the needs of individuals with a mental illness within an increasingly technology based environment. This requires the development and evaluation of educational strategies that immerse students in web-based learning. This article presents an overview of a hybrid teaching design that includes classroom teaching and asynchronous threaded discussion in a teaching module in an undergraduate psychiatric nursing course. Evaluation of student preferences, advantages and disadvantages, and learning, as well as qualitative evaluation of students' description of critical thinking, supports the value of online teaching in psychiatric nursing education. PMID- 16308128 TI - A comparative study of postpartum depression in abused and non-abused women. AB - Because of a rise in postpartum depression (PPD), the U.S. Department of Health and Human Services recently indicated a need for more studies of possible risk factors for PPD. PPD can continue for months or years after birth and has consequences for the family as a whole. Past research has not investigated the relationships between abuse, whether physical, sexual, or emotional, and PPD. The aim of this pilot study was to compare the prevalence of PPD among abused and non abused women during the first 4 months after birth. Although 89 women initially enrolled in the study, only 50 participants completed the 1-week data collection. Of these 50, 30% reported previous or current physical or sexual abuse. Of these 50 participants, 22 dropped out during the 4-month-long study, resulting in a final sample size of 28. Abused women were significantly more depressed at 1 week and 2 and 3 months postpartum than the non-abused group until their depression scores at entry into the study and 1 week were statistically controlled. An unexpected finding was the prevalence of depression at 1 week after birth, indicative of unresolved prenatal depression, "baby blues," or true PPD. In addition, emotional abuse was reported by both groups and was significantly higher for the previously abused women. Although sample size and high attrition limit generalization, there was no significant difference in the prevalence of PPD between the two groups. Future research should explore the relationships between abuse and depression with larger samples and include pregnant subjects. PMID- 16308129 TI - Prospective, randomized, controlled, single-blind trial of the costs and consequences of systematic nutrition team follow-up over 12 mo after percutaneous endoscopic gastrostomy. AB - OBJECTIVES: We assessed the economic and clinical implications of systematic long term nutrition team follow-up of patients after percutaneous endoscopic gastrostomy. METHODS: We designed a prospective, randomized, controlled, single blind trial in a large district hospital and its catchment area. All adult patients referred for a gastrostomy were eligible and randomized into two groups. The intervention group had regular follow-up by the nutrition team (weekly in hospital, monthly after discharge) with appropriate support and advice for patient, carer, and primary care professionals. The control group had no specific nutrition team input (as is often current practice). Endpoints to the study were 12 mo, elective removal of tube, or death. The primary outcome was total health care costs. Secondary outcomes were complications, length of stay, readmissions, nutritional status, and quality of life. RESULTS: One hundred twelve patients were recruited. Eleven died before the start of the trial, leaving 47 in the intervention group and 54 in the control group. They were well matched for age, sex, and underlying diagnosis. Overall, the health care costs were 13,330 sterlings per patient in the intervention group compared with 16,858 pound sterlings in the control group (two-tailed, P = 0.27), a saving of 21% per patient. The intervention group had shorter lengths of stay, fewer and briefer readmissions, earlier removal of gastrostomy (where appropriate), shorter duration of feeding, and less demand for general practitioners and district nurse inputs. Nutritional status and quality of life were similar. CONCLUSIONS: Regular systematic nutrition team follow-up for gastrostomy-fed patients does not increase costs and may improve quality of care. PMID- 16308130 TI - Hospital resources consumed for surgical morbidity: effects of preoperative arginine and omega-3 fatty acid supplementation on costs. AB - OBJECTIVE: Costs related to postoperative complications continue to be a major burden on any health care system. The aim of the present study was to calculate hospital costs for postoperative complications and to evaluate whether preoperative supplementation with omega-3 fatty acids and arginine (specialized diet) might lead to cost savings in patient care. METHODS: Blind analysis of costs performed on data gathered from a randomized clinical trial carried out on 305 patients with gastrointestinal cancer showed that an oral preoperative specialized diet decreased postoperative morbidity compared with conventional treatment (no supplementation). Estimates of complication costs were based on resources used for treatment and on additional length of hospital stay. Cost comparison and cost-effectiveness analyses were then carried out. RESULTS: The mean cost of postoperative complications was 4492 pounds sterlings. The greatest amount of resources was consumed by 19 anastomotic leaks (159,803 pounds sterlings), 18 abdominal abscesses (112,921 pounds sterlings), and 18 pancreatic fistulae (106,516 pounds sterlings). The mean costs per complication were 6178 pounds sterlings in the conventional group and 4639 pounds sterlings in the preoperative group (P = 0.05). The mean total costs of patients with complications were 10,494 pounds sterlings in the conventional group and 8793 pounds sterlings in the preoperative group. The mean cost per randomized patient was 3122 pounds sterlings in the conventional group versus 1872 pounds sterlings in the preoperative group (P = 0.04). Effectiveness values were 50.0% in the conventional group and 62.8% in the preoperative group (P = 0.03). Total costs consumed 93% of the diagnosis-related group reimbursement rate in the conventional group and 78% in the preoperative group. CONCLUSIONS: The costs of postoperative morbidity consumed a large amount of the diagnosis-related group reimbursement rate. Preoperative supplementation with the specialized diet appears to be a cost-effective treatment. PMID- 16308131 TI - Maternal plasma adiponectin concentrations at 24 to 31 weeks of gestation: negative association with gestational diabetes mellitus. AB - OBJECTIVE: Adiponectin is an adipocyte-derived hormone with antidiabetic, antiatherosclerotic, and antiinflammatory properties. This study investigated the relations between maternal adiponectin concentration and gestational diabetes mellitus (GDM) and other metabolic parameters during midpregnancy. METHODS: Two hour 75-g oral glucose tolerance tests were performed in 253 pregnant women at 24 to 31 wk of gestation. Two hundred nineteen who had normal glucose tolerance (NGT) and 34 women who had GDM and their newborns were investigated. Fasting maternal blood samples were drawn to determine plasma concentrations of adiponectin, glucose, insulin, C-peptide, free fatty acid, and blood lipids. Blood samples at 1 and 2 h after an oral glucose load were obtained to measure plasma glucose, insulin, and C-peptide concentrations. RESULTS: Plasma adiponectin concentrations were significantly lower in women who had GDM than in those who had NGT (P = 0.014). Maternal age, body mass index (before pregnancy and at blood collection), and plasma level of free fatty acid were significantly greater in those who had GDM than in those who had NGT. Logistic regression analysis showed that maternal adiponectin level and GDM were significantly correlated (P = 0.043), but that the correlation became weaker (P = 0.116) after adjusting for maternal body mass index and plasma level of free fatty acid before pregnancy. In the NGT group, maternal adiponectin concentrations were significantly negatively correlated with plasma fasting insulin, fasting C peptide, fasting C-peptide/fasting glucose ratio, 2-h glucose, triacylglycerol, and maternal body mass index and positively correlated with high-density lipoprotein cholesterol concentration. In the GDM group, maternal adiponectin level was negatively correlated with neonatal birth weight. CONCLUSIONS: Midpregnancy hypoadiponectinemia may be associated with a higher risk of GDM. PMID- 16308132 TI - Geographical inequalities in nutrient status and risk of malnutrition among English people aged 65 y and older. AB - OBJECTIVE: Geographical inequalities in health continue to be a problem within developed countries. This study investigated whether there were north-south geographical inequalities among older people living in England with respect to risk of protein-energy malnutrition and status of nutrients, particularly those derived from fruit and vegetables. METHODS: A secondary analysis of data collected prospectively by the National Diet and Nutrition Survey of people aged 65 y and older was undertaken to assess geographical prevalence of risk of protein-energy malnutrition (1155 subjects) and nutrient status (881 to 1046 subjects). RESULTS: A north-south gradient was found in risk of protein-energy malnutrition (19.4%, 12.3%, and 11.2% in the northern, central, and southern regions, respectively; P = 0.013, P for trend = 0.002). This was accompanied by a north-south gradient in the status of vitamin C (30, 38, and 46 micromol/L in the respective regions, P < 0.001), which was associated with deficiency (<11 micromol/L) in a third of subjects in the northern region, a range of carotenoids (P = 0.023 to <0.001), vitamin D (P < 0.001), and selenium (P < 0.001). These inequalities were accompanied by gradients in indices of health status and socioeconomic status, which could account only partly for the "geographical" inequalities. Circulating vitamin C and carotenoid concentrations were related to the intake of fruit and vegetables. CONCLUSIONS: This study suggests there is a north-south divide in the risk of protein-energy malnutrition and a range of nutrients, which have been implicated in the development of common chronic diseases. PMID- 16308133 TI - Prospective study of resting energy expenditure changes in head and neck cancer patients treated with chemoradiotherapy measured by indirect calorimetry. AB - OBJECTIVE: The prevalence of malnutrition is high in patients with head and neck cancer due to tumor location and coadjuvant treatment. We studied changes in resting energy expenditure (REE) during treatment with chemoradiotherapy and compared the measured REE by indirect calorimetry (IC) with the value estimated by the Harris-Benedict (HB) formula. METHODS: Eighteen patients with head and neck cancer (15 men and 3 women, mean age 57 +/- 10.7 y, age range 30-71 y) entered the study. All patients were treated with radiotherapy (70.8 +/- 1 Gy, range 70-72) and received 37.4 +/- 3.5 fractions (range 32-42) and concurrent chemotherapy with cisplatinum (absolute doses of 400 to 1000 mg). Nutrition assessment included anthropometry (body mass index, triceps skinfold thickness, subscapular skinfold thickness, midarm circumference, and midarm muscle circumference) and tetrapolar bioimpedance (Holtain BC). The IC (kcal/24 h; Deltatrac II MBM-200) was performed after an overnight fast. Measurements were done before treatment, at weeks 2, 4, and 6 of treatment, at the end of treatment, and 2 wk after treatment. RESULTS: Body mass index decreased during treatment from 24.7 +/- 4.4 kg/m(2) (range 16.9-31.4) to 22.3 +/- 4.2 kg/m(2) (range 15.1-29.6). REE (kcal/24 h) changed significantly during treatment (IC P < 0.05, HB formula P < 0.001). REE measured by IC appeared as a U-shaped curve, but REE estimated by the HB formula decreased during treatment. The HB underestimated REE measurements compared with IC. These differences were statistically significant before treatment, at the end of treatment, and 2 wk after treatment (P < 0.05) and showed a limited clinical agreement with the Bland-Altman method. CONCLUSIONS: REE measured by IC significantly changed during chemoradiotherapy. It was higher before treatment, at the end of treatment, and 2 wk after treatment. The HB formula underestimated REE in these patients. IC is a suitable method for measuring REE in this cohort of patients. PMID- 16308134 TI - Vitamin B6 intake by Koreans should be based on sufficient amount and a variety of food sources. AB - OBJECTIVES: This study estimated dietary vitamin B6 intake and its food sources in young Koreans by using a modified Korean vitamin B6 database. METHODS: Dietary vitamin B6 intake and food sources were estimated from a 3-d recall method for 294 Korean college undergraduates (149 men and 145 women, age range 17 approximately 25 y) who had no health problems. Food portions were estimated by using standard household measurements and published average portions. RESULTS: Average daily intakes of vitamin B6 were 1.57 mg/d in men and 1.44 mg/d in women. Average consumptions of vitamin B6 were 104% of the Korean recommended dietary allowance in men for vitamin B6 and 102% in women. However, 37% of men and 32% of women subjects consumed less than 75% of the Korean recommended dietary allowance for vitamin B6. Major dietary sources of vitamin B6 for Korean young men were pork, rice, onion, potatoes, garlic, mackerel, spinach, egg, carrot, and chicken. Major dietary sources of vitamin B6 for young Korean women were rice, pork, kimchi, beef, bean sprout, potatoes, dumplings, and onions. As for major dietary sources of vitamin B6, the top 20 foods provided nearly 89% of total vitamin B6 in men and 70% of total vitamin B6 in women. CONCLUSIONS: Intake of vitamin B6 by young Koreans was adequate in general, but the supply of vitamin B6 for Koreans should be based on sufficient amounts and specific food sources because fewer foods contribute to vitamin B6 intake of Koreans and these foods are mainly of plant origin and less bioavailable. PMID- 16308135 TI - Increase in bone mineral density through oral administration of shark gelatin to ovariectomized rats. AB - OBJECTIVE: The incidence of bovine spongiform encephalopathy has resulted in marine collagen hydrolysate (low-molecular-weight gelatin) being sold as supplements and cosmetics in Japan. Shark skin collagen is one of the important sources of marine collagen. We examined the effect of shark skin gelatin in an osteoporosis model animal. METHODS: Shark skin gelatin was orally administered to ovariectomized rats with a low-protein diet. Bone mineral density of the right femur was measured. Collagen and glycosaminoglycan in the tibial end were extracted and analyzed by western blotting and cellulose acetate membrane electrophoresis, respectively. RESULTS: Administering collagen to the ovariectomized rats resulted in the bone mineral density of the femur epiphysis being higher than that in the sham-operated rats. The contents of type I collagen and glycosaminoglycan in the epiphysis were increased by administering shark skin gelatin. CONCLUSIONS: These results indicated that shark skin gelatin would be useful as a dietary supplement for treating osteoporosis. PMID- 16308136 TI - Cost savings of an adult hospital nutrition support team. AB - OBJECTIVES: A hospital-based nutrition support team (NST) may need to demonstrate cost savings and quality benefits. The primary aim of this study was to determine whether an NST could show tangible cost savings (equipment, investigations, and medication costs) from managing patients considered for parenteral nutrition (PN). Secondary aims related to the quality issues of placement of PN catheters, catheter-related sepsis (CRS), duration of parenteral nutrition, and mortality. METHODS: An NST was formed in 1999 and worked in all adult areas of a university hospital (Leicester Royal Infirmary). Comparative data about all patients given PN were collected for 2 consecutive years (a retrospective pre-NST year and a prospective NST year). RESULTS: In the pre-NST year there were 82 PN episodes (54 patients), 665 PN days, and a CRS rate of 71% (seven infections/100 PN days). In the NST year, there were 133 referrals for PN but only 78 PN episodes (75 patients, 59% of referrals), 752 PN days, and a decreased overall CRS rate of 29% (three infections/100 PN days, P < 0.05) but a rate of 7% (0.6 infection/100 PN days) in the final 3 mo of the NST year. Tangible cost savings for the NST year were derived from 55 avoided PN episodes (42741 pounds sterlings) and 35 avoided CRS episodes (7974 pounds sterlings). Thirty-nine percent of PN catheters were inserted by the NST with no insertion-related complications. Competency-based training of ward nursing staff decreased the CRS rate. Mean duration of PN increased from 8 to 10 d (P not significant). In-hospital mortality for patients who had PN was 23 of 54 (43%) in the pre-NST year compared with 18 of 75 (24%) in the NST year (P < 0.05). CONCLUSIONS: Although the number of PN days increased with an NST, tangible cost savings of 50715 pounds sterlings were demonstrated within the NST year by avoided PN episodes and a decreased incidence of CRS. These savings justify the salaries of a nutrition nurse specialist and a senior dietitian. PMID- 16308137 TI - Effect of glutamine on cell adhesion molecule expression and leukocyte transmigration in endothelial cells stimulated by preeclamptic plasma. AB - OBJECTIVE: This study analyzed plasma glutamine (GLN) concentrations in women with preeclampsia. Also, in an in vitro study we evaluated whether GLN concentration was related to surface molecule expressions on endothelial cells (ECs) and polymorphonuclear neutrophils (PMNs) and the transendothelial migration of PMNs through ECs stimulated by preeclamptic plasma. METHODS: Blood samples were collected from 20 women with preeclampsia and 15 normal pregnant women for plasma GLN analysis. In the in vitro study, human umbilical vein endothelial cells and PMNs were treated with different concentrations (0, 300, 500, and 1000 microM) of GLN for 24 h. After that, we stimulated human umbilical vein endothelial cells for 3 h with plasma from patients with preeclampsia, and PMNs were allowed to transmigrate through ECs for 2 h. EC surface expressions of cellular adhesion molecules (CAMs) and integrin (CD11b) interleukin-8 (IL-8) receptor expressions on PMNs were measured by flow cytometry. The transendothelial migration of PMNs through ECs was also analyzed. RESULTS: Women with preeclampsia exhibited significantly lower plasma GLN concentrations than did normal pregnant women. The in vitro study showed that, compared with normal plasma, CAM expressions on human umbilical vein endothelial cells and PMNs were increased when preeclamptic plasma was stimulated. Among the groups with preeclamptic plasma stimulation, intracellular CAM-1 expression on ECs and CD11b and IL-8 receptor expressions on PMNs were lower with 500 and 1000 microM than with 300 microM of GLN. IL-8 production from ECs and PMNs was also lower with 500 and 1000 microM than with 300 microM of GLN. PMN transmigration was significantly higher with 300 microM of GLN than with the other GLN concentrations. CONCLUSIONS: Plasma GLN is depleted in women with preeclampsia. The result of this in vitro study showed that ECs and PMNs were activated after preeclamptic plasma stimulation. A low GLN concentration resulted in greater CAM expression and greater transendothelial migration of neutrophils. GLN administration at levels similar to or higher than physiologic concentrations decreased IL-8 and CAM expressions, and PMN transmigration decreased after stimulation with preeclamptic plasma. PMID- 16308138 TI - Induction of apoptosis by the aqueous extract of Rubus coreanum in HT-29 human colon cancer cells. AB - OBJECTIVES: The incompletely ripened fruit of Rubus coreanum (IRFRC) has been used in traditional herbal medicine to manage various diseases. To explore the possibility that IRFRC has chemopreventive effects, we examined whether or not extracts of IRFRC inhibits HT-29 cell growth and explored the mechanism for this effect. METHODS: We cultured HT-29 cells in the presence of the aqueous or ethanol extract of IRFRC. DNA synthesis was estimated by 5-bromo-2'-deoxyuridine incorporation. We measured apoptosis using a DNA fragmentation assay and Annexin V staining. We used western blot analyses to determine the cleavage of caspases and poly (adenosine diphosphate-ribose) polymerase. RESULTS: Aqueous extract of IRFRC substantially inhibited viable HT-29 cell number in a dose-dependent manner, whereas ethanol extract had only a minimal effect. Aqueous extract inhibited DNA synthesis and induced apoptosis of HT-29 cells in a dose-dependent manner. Aqueous extract induced cleavage of caspase-3, -7, and -9 and induced the activity of caspase-3 and cleavage of poly (adenosine diphosphate-ribose) polymerase. CONCLUSIONS: We have shown that aqueous extract of IRFRC inhibits cell proliferation and stimulates apoptosis in HT-29 cells, and that this may be mediated by its ability to activate the caspase-3 pathway. It remains to be determined whether the aqueous extract of IRFRC has chemopreventive activities in animal models. PMID- 16308139 TI - Credit where credit is due. PMID- 16308140 TI - Stunted girls have greater subcutaneous fat deposits: what type of intervention can improve the health of stunted children? PMID- 16308141 TI - Why albumin now? Let's think about albumin measurement methods, the definition of hypoalbuminemia, and microheterogeneity. PMID- 16308142 TI - Bioavailability of cellobiose and other non-digestible and/or non-absorbable sugar substitutes and related topics. PMID- 16308143 TI - Polycystic ovary syndrome, a pathway to type 2 diabetes. PMID- 16308144 TI - Food culture may be a risk factor for colorectal cancer among Argentineans: the human history of this research. PMID- 16308145 TI - Fasting breath hydrogen concentrations in patients with short bowel syndrome with colon in continuity before and after antibiotic therapy. PMID- 16308146 TI - Conjugated linoleic acid, obesity, and insulin resistance: waiting for the day of liberation from chronic disease. PMID- 16308147 TI - Isolated perfused liver model: the rat and the guinea pig compared. PMID- 16308148 TI - Discourse on diabetes, diet, and cell death: steps toward a purpose. PMID- 16308149 TI - Beyond the bag horizon. PMID- 16308151 TI - Gene discovery in neuropharmacological and behavioral studies using Affymetrix microarray data. AB - We describe methods and software tools for doing data analysis based on Affymetrix microarray data, emphasizing often neglected issues. In our experience with neuroscience studies, experimental design and quality assessment are vital. We also describe in detail the pre-processing methods we have found useful for Affymetrix data. Finally, we summarize the statistical literature and describe some pitfalls in the post-processing analysis. PMID- 16308152 TI - RNA amplification strategies for small sample populations. AB - Advances in high throughput cloning strategies have led to sequencing of the human genome as well as progress in the sequencing of the genome of several other species. Consequently, the field of molecular genetics is blossoming into a multidisciplinary entity that is revolutionizing the way researchers evaluate a myriad of critical concepts such as development, homeostasis, and disease pathogenesis. There is tremendous interest in the quantitative assessment of tissue-specific expression of both newly identified and well characterized specific genes and proteins. At present, an ideal approach is to assess gene expression in single elements recorded physiologically in living preparations or by immunocytochemical or histochemical methods in fixed cells in vitro or in vivo. The quantity of RNA harvested from individual cells is not sufficient for standard RNA extraction methods. Therefore, exponential polymerase-chain reaction based analyses, and linear RNA amplification including amplified antisense RNA amplification and a newly developed terminal continuation RNA amplification methodology have been developed for use in combination with microdissection procedures and cDNA/oligonucleotide microarray platforms. RNA amplification is a series of intricate procedures to amplify genetic signals from minute quantities of starting materials for microarray analysis and other downstream genetic methodologies. RNA amplification procedures effectively generate quantities of RNA through in vitro transcription. The present report illustrates practical usage of RNA amplification technologies within the context of regional, population cell, and single cell analyses in the brain. PMID- 16308153 TI - Production of custom microarrays for neuroscience research. AB - Microarray chips produced by commercial vendors and academic laboratories are mostly generic in nature to facilitate wide applicability. With the sequencing of the human, mouse, and rat genomes, the thrust is to expand clone and oligonucleotide sets and increase the number of genes represented on a particular array. This is appropriate for discovery based investigations where microarray technology has been successfully utilized. However, array technology can also be employed to perform hypothesis based studies if optimized chips can be produced with relevant content. Existing array technology available at core facilities can be effectively utilized to produce a custom microarrays with genes that are most relevant to the research interests of individual investigators or research groups for use as a standard molecular tool. The power of this technology can be harnessed to further our understanding of specific biological problems without involvement in extensive data mining and analysis. The custom microarray approach is presented with procedural details for design and production in the context of neurobiological investigations. PMID- 16308154 TI - A microarray data analysis framework for postmortem tissues. AB - This paper will give a complete methodological approach to the processing of oligonucleotide microarray data from postmortem tissue, particularly brain matter. Attention will be drawn to each of the important stages in the process; specifically the quality control, gene expression value calculation, multiple hypothesis testing and correlation analyses. We shall initially discuss the theoretical foundations of each individual method and subsequently apply the ensemble to a sample data set to illustrate and visualise important points. PMID- 16308155 TI - True and false discovery in DNA microarray experiments: transcriptome changes in the hippocampus of presenilin 1 mutant mice. AB - In transcriptome profiling experiments using DNA microarrays, it is critical to maximize putatively true data discovery while keeping the false discovery rate at acceptable levels. Using previously published and verified transcriptome datasets of mice with genetically altered PS1 physiology, we present a simple, robust, and system-specific assessment of type I and type II errors in two independent microarray experimental series. We provide evidence to suggest that for maximizing true discovery and minimizing false discovery, statistical criteria alone are inferior to statistical significance plus magnitude of change criteria. Furthermore, we found that, regardless of the exact criteria used for determining differential expression, different data extraction protocols give rise to different discovery and false discovery rates. In addition, a large proportion of expression differences were both dataset and analytical approach dependent. The data assessment methods presented and discussed in this manuscript can be easily carried out on any microarray dataset using basic spreadsheet functions as the only tool needed. Finally, we provide an in-depth analysis of the hippocampal transcriptome of DeltaE9 hPS1 transgenic mice and mice with a conditional ablation of the PS1 gene. PMID- 16308156 TI - Convergent functional genomics: a Bayesian candidate gene identification approach for complex disorders. AB - Identifying genes involved in complex neuropsychiatric disorders through classic human genetic approaches has proven difficult. To overcome that barrier, we have developed a translational approach called Convergent Functional Genomics (CFG), which cross-matches animal model microarray gene expression data with human genetic linkage data as well as human postmortem brain data and biological role data, as a Bayesian way of cross-validating findings and reducing uncertainty. Our approach produces a short list of high probability candidate genes out of the hundreds of genes changed in microarray datasets and the hundreds of genes present in a linkage peak chromosomal area. These genes can then be prioritized, pursued, and validated in an individual fashion using: (1) human candidate gene association studies and (2) cell culture and mouse transgenic models. Further bioinformatics analysis of groups of genes identified through CFG leads to insights into pathways and mechanisms that may be involved in the pathophysiology of the illness studied. This simple but powerful approach is likely generalizable to other complex, non-neuropsychiatric disorders, for which good animal models, as well as good human genetic linkage datasets and human target tissue gene expression datasets exist. PMID- 16308157 TI - Integrated expressional analysis: application to the drug discovery process. AB - Microarray technology enables high-throughput testing of gene expression to investigate various neuroscience related questions. This in turn creates a demand for scalable methods to confirm microarray results and the opportunity to use this information to discover and test novel pathways and therapeutic applications. Discovery of new central nervous system (CNS) treatments requires a comprehensive understanding of multiple aspects including the biology of a target, the pathophysiology of a disease/disorder, and the selection of successful lead compounds as well as efficient biomarker and drug disposition strategies such as absorption (how a drug is absorbed), distribution (how a drug spreads through an organism), metabolism (chemical conversion of a drug, if any, and into which substances), and elimination (how is a drug eliminated) (ADME). Understanding of the toxicity is also of paramount importance. These approaches, in turn, require novel high-content integrative assay technologies that provide thorough information about changes in cell biology. To increase efficiency of profiling, characterization, and validation, we established a new screening strategy that combines high-content image-based testing on Array Scan (Cellomics) with a confocal system and the multiplexed TaqMan RT-PCR method for quantitative mRNA expression analysis. This approach could serve as an interface between high throughput microarray testing and specific application of markers discovered in the course of a microarray experiment. Markers could pinpoint activation or inhibition of a molecular pathway related, for instance, to neuronal viability. We demonstrate the successful testing of the same cell population in an image based translocational assay followed by poly(A) mRNA capture and multiplexed single tube RT-PCR. In addition, Ciphergen ProteinChip analysis can be performed on the supernatant, thus allowing significant complementarity in the data output and interpretation by also including the capture and initial analysis of proteins in the integrative approach presented. We have determined various conditions including the number of cells, RT and PCR optimization, which are necessary for successful detection and consequent assay integration. We also show the successful convergence of various different approaches and multiplexing of different targets within a single real-time PCR tube. This novel integrative technological approach has utility for CNS drug discovery, target and biomarker identification, selection and characterization as well as for the study of toxicity- and adverse event-associated molecular mechanisms. PMID- 16308158 TI - Fascin expression in 90 patients with glioblastoma multiforme. AB - Fascin is a protein that serves to aggregate F actin into bundles that rearrange the cytoskeleton and promote cellular motility. Fascin has been linked to the invasive behavior of some tumors. Fascin immunohistochemical analysis was performed in 90 glioblastoma multiforme, including 53 males and 37 females (mean age, 58.3 years). All patients had tumors that demonstrated positive fascin staining. Nineteen tumors showed more than 75% positive staining tumor cells, 14 tumors had more than 50% to 75% staining, 23 tumors had more than 25% to 50% staining, 26 tumors had more than 5% to 25% staining, and 8 tumors had less than 5% staining. In comparison, 9 of 11 low-grade astrocytomas had 50% or less staining for fascin. Eight of 10 anaplastic astrocytomas had more than 50% fascin staining. All gliomas studied expressed fascin by immunohistochemistry. Higher grade tumors generally expressed a greater degree of fascin staining. There was no obvious correlation with the extent of staining and survival among glioblastoma multiforme. Fascin may play a role in tumor cell infiltration. PMID- 16308159 TI - Expression of hyaluronic acid and its receptors, CD44s and CD44v6, in normal, hyperplastic, and neoplastic endometrium. AB - The interaction between epithelial tumor cells and their surrounding stroma is important in tumor progression and metastasis. This is accomplished through a number of transmembrane receptors that interact with stromal extracellular matrix molecules. One of these receptors, CD44, binds to extracellular matrix component hyaluronic acid (HA). The purpose of this study was to evaluate the significance of HA, CD44s, and CD44v6 in benign, hyperplastic, atypical, and malignant endometrial epithelia. Archival paraffin-embedded cell blocks from proliferative endometrium (n = 11), secretory endometrium (n = 12), simple hyperplasia (n = 13), complex hyperplasia without atypia (n = 9), complex hyperplasia with atypia (n = 17), and adenocarcinoma (n = 21) were stained for HA, CD44s, and CD44v6. HA was detected throughout the normal menstrual cycle but was more intense during the secretory phase. Only during the secretory phase was CD44s expressed in the stromal cells in 11 cases (92%), whereas CD44v6 was detected in glandular epithelium in 9 (75%). CD44s was expressed in the glandular epithelium in 2 (15%) cases of simple hyperplasia, 4 (44%) of complex hyperplasia without atypia, 14 (82%) of complex hyperplasia with atypia, and in 16 (76%) of adenocarcinoma. CD44v6 was expressed in the glandular epithelium in 1 (11%) case of complex hyperplasia without atypia, 17 (100%) cases of complex hyperplasia with atypia, and in 18 (86%) cases of adenocarcinoma, but in none of the cases of simple hyperplasia. The endometrial stromal cells expressed CD44v6 in 1 (8%) case of simple hyperplasia, 6 (67%) of complex hyperplasia without atypia, 8 (47%) of complex hyperplasia with atypia, and in 3 (14%) of adenocarcinoma. We concluded that in the normal menstrual cycle, the timing of peak staining of HA and CD44s in the stroma and the up-regulation of CD44v6 in secretory glands are coincident with the period in which the endometrium is most receptive to embryo implantation. HA is more abundant in the stroma adjacent to the tumor, suggesting that interactions between tumor cells and stromal HA promote tumorigenesis. With progression from hyperplasia and with increasing atypia to adenocarcinoma, levels of stromal HA, glandular CD44v6, and glandular and stromal CD44s all increase. Thus, HA and CD44 are both involved in the development and progression of endometrial cancer. PMID- 16308160 TI - Life-threatening hemoptysis caused by chronic idiopathic pulmonary hilar fibrosis with unilateral pulmonary vein occlusion. AB - We present the case of a patient who has a life-threatening hemoptysis caused by occlusion of the right pulmonary vein owing to external compression by excessive fibrous tissue. Because the patient's lung was essentially nonfunctional and hemoptysis persisted, we performed a pneumonectomy on her. Pathological analysis of the lung showed severe hypertensive changes in the arterial and venous microvasculature of the lung. Parenchymal-pleural to intercostal systemic venous connections had developed, facilitating venous drainage of the lung circulation. Upon review of the patient's history and the pathological and radiological evidence, we concluded that the pathological changes were caused by a variant of mediastinal fibrosis termed chronic idiopathic pulmonary hilar fibrosis, of which only a few cases have been described in the literature. A detailed case history and review of the literature are presented. PMID- 16308161 TI - Laryngotracheoesophageal cleft type 3 and double outlet right ventricle: unique combination. AB - The laryngotracheoesophageal cleft is marked by the absence of the anatomic separation of the esophagus and the larynx. The cleft functionally can be restricted to involve the interarytenoid musculature (type 1), cricoid involvement (type 2), the proximal laryngotracheoesophagus (type 3), or missing of the whole tracheoesophageal septum (type 4). Association with significant congenital heart disease is uncommon. In this report, we are presenting a 2-week old preterm girl presented with frequent aspiration pneumonitis, cyanosis, and heart murmur. Mechanical ventilation was extremely difficult because of endotracheal tube air leak and excessive secretions. Endoscopic evaluation revealed type 3 laryngotracheoesophageal cleft. Echocardiogram revealed double outlet right ventricle with subpulmonic ventricular septal defect. Such unique combination with severe airway disease and complex congenital heart disease was thought to constitute very high morbimortality if intervention is contemplated. It was elected to proceed with compassionate care. PMID- 16308162 TI - Subscapular elastofibroma intermingled with adipose tissue: variant type of elastofibroma or lipoma? AB - Elastofibroma is a rare entity most commonly seen in subscapular region. Elastofibrolipoma was first described by De Nictolis et al (Am J Surg Pathol. 1995;19:364-367) as a capsulated mass, composed of mature fat, abnormal elastic fibers, and connective tissue, at the anterior mediastinum. A 57-year-old woman with a periscapular pain and a mass at the left subscapular region was admitted at the department of thoracic surgery. Histologic sections of the well circumscribed 6.5 x 5 x 3.5-cm rubbery mass revealed a collagenous stroma containing eosinophilic globules and fragmented fibers intermixed with mature adipose tissue surrounded by a thin fibrous capsule. The elastic nature of these eosinophilic fibers and globules was confirmed with Verhoeff's elastic stain. The lesion was diagnosed as elastofibrolipoma, a recently recognized lesion with an unclear origin. Here we discuss if it is a variant of elastofibroma or lipoma. PMID- 16308163 TI - C-Kit-positive metastatic malignant pigmented clear-cell epithelioid tumor arising from the kidney in a child without tuberous sclerosis. AB - We report the first pediatric case of malignant pigmented epithelioid clear-cell tumor arising from kidney; the lesion occurred in a 12-year-old girl without tuberous sclerosis. The tumor was widely metastatic to the retroperitoneum and chest, and the patient died of the disease 9 months after diagnosis, despite active chemotherapy. Pigmented epithelioid clear-cell tumor of the kidney is a rare variant of epithelioid angiomyolipoma and a member of the family of perivascular epithelioid cell tumors (PEComas). The tumor demonstrated overlapping features between clear-cell sugar tumor and epithelioid variant of angiomyolipoma. Tumor cells were positive for HMB-45 expression, negative for any evidence of muscular differentiation, and contained melanin pigment and premelanosomes in the cytoplasm. Diffuse C-Kit (CD117) positivity was identified throughout the tumor. This is the first report of C-Kit-positive malignant PEComas. PMID- 16308164 TI - Low-grade metaplastic adenocarcinoma (carcinosarcoma) of the uterus: report of an unusual case. AB - An unusual low-grade metaplastic carcinoma (mullerian carcinosarcoma) of the uterine corpus occurring in a 45-year-old woman is presented. The tumor was polypoid and showed an initial invasion into the myometrium. The epithelial component was composed of low-grade endometrioid glands that focally showed an unusual oxyphilic change of the cytoplasm. These cells contained only a small amount of mitochondria detected with antimitochondrial antibody. The stromal component contained small round cells of bland appearance and appeared to arise from epithelial cells through a metaplastic process, as a gradual transition between epithelial and stromal cells was often seen. An invasive marginal zone of the tumor showed endometrioid adenocarcinoma pattern without any stromal component. The proliferative activity generally was low. Interestingly, it was clearly accentuated in the epithelial component especially in the peripheral invasive areas, whereas it was very low in the stromal component. The course of disease was indolent. Low-grade mullerian carcinosarcoma is to be distinguished from other biphasic mullerian tumors. PMID- 16308165 TI - Extranodal lymphomas of the head and neck. AB - Malignant lymphomas represent approximately 5% of all malignant neoplasms of the head and neck and may involve nodal or extranodal sites. Nodal head and neck lymphomas are similar to other nodal sites and are not further reviewed here. The head and neck region is the second most frequent anatomical site of extranodal lymphomas (after the gastrointestinal tract). Most are non-Hodgkin's lymphomas of B-cell lineage, and overall diffuse large B-cell lymphoma is the most common type. Hodgkin's lymphoma rarely occurs in extranodal sites. Other hematologic neoplasms that commonly involve extranodal sites of the head and neck are also discussed. In this review, we begin by discussing lymphomas involving the head and neck according to anatomical site. Then we discuss specifically the pathological findings of extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue, plasmablastic lymphoma, extramedullary plasmacytoma, and extranodal natural killer/T-cell lymphoma of nasal type. PMID- 16308167 TI - Multi-agency voices: a thematic analysis of multi-agency working practices within the setting of a Child and Adolescent Mental Health Service. AB - This qualitative study aims to explore the types and purpose of discourse emerging when professionals from a Child and Adolescent Mental Health Service meet with professionals from other agencies to discuss cases. The aim reflects current political and contextual agendas influencing agencies to work more closely together and obstacles to achieving this goal highlighted in the literature such as the need for agencies to develop a common language to discuss children of concern. Data were collected through eight audio-taped meetings involving CAMHS and members of other agencies such as social services and education. A thematic analysis identified nine themes, defined according to their discourse type, including: single agency discourse; case complexity discourse and multi-agency discourse. Results indicate that agencies are hindered from working more closely together and developing a common language for use in multi-agency meetings, because they tend to have different understandings of the terminology used and a common consensus about language and meaning is not usually negotiated within the meeting. There is a need for greater awareness amongst staff from different professional groups and agencies that meaning given by one group will often need to be clarified by others and assumptions about common understandings should not be made. PMID- 16308168 TI - The significance of a collaborative practice model in delivering care to chronically ill patients: a case study of managing diabetes mellitus in a primary health care center. AB - Non-communicable diseases, especially diabetes mellitus type two (DM2) constitute major health problems in Lebanon that have an adverse impact on health and health resources. Collaborative practice interventions may improve quality care of DM2 and reduce or delay complications. The purpose of this paper was to evaluate the impact of collaborative practice on the quality and cost of effective care for diabetic patients in a primary health care center. A chart audit review of 375 diabetic patients attending an inner city health center in Beirut (Lebanon) was conducted after three and a half years of collaborative practice intervention, which included guidelines for an interdisciplinary health team. Evaluation of the impact of collaborative practice was conducted on the process and outcome of care. The results indicated a high level of enthusiasm, support and the development of team spirit at the process level. At the outcome level there was improvement in documentation, increase in patient recruitment, increase in continuity of care, improvement of glycemic control and decreased cost. In conclusion collaborative practice interventions improved process and outcome variables for diabetic patients. It is suggested that this model could be developed for use in the care of other chronic diseases. PMID- 16308169 TI - Developing an interprofessional learning culture in primary care. AB - This paper discusses the concept of interprofessional learning based on empirical data from an evaluation of an interprofessional learning project that was set in a British primary health care centre. A process evaluation methodology was chosen to collect the data using semi-structured interviews and focus groups with stakeholders and staff to gather their experiences and views of the project and documentary data from records written over the lifetime of the project. The paper argues that an interprofessional learning culture requires time to become embedded in everyday practice and to achieve such a culture, shared values, aims and clear communication are essential. The data suggest that there is a need to recognize responsibility for one's own learning as individuals as well as learning as teams of work colleagues if interprofessional learning is to be successful. However, even when these pre-requisites of interprofessional learning are agreed and acknowledged openly in the workplace, participants in the development of a learning culture need to recognize that there are structural controls which influence and constrain such developments which are external to participants and beyond their immediate control. PMID- 16308170 TI - School staff members' and occupational health nurses' evaluation of the promotion of occupational well-being - with good planning to better practice. AB - This paper describes occupational health nurses' and school staff members' experiences of work ability maintenance carried out at schools and the development of ideas to promote the collaboration between occupational health care and school community and to enhance work ability maintenance. The data were collected in a baseline situation of an action research project to further plan and develop activities. The data were analysed by inductive content analysis. According to the results, school community staff members' work ability maintenance focused on the employee, the school community, school work and the working conditions as well as professional competence. The practical actions involved some individual and school-specific variation, focusing on actions to maintain individual and physical work ability. It seems that the collaboration between the school community and occupational health service for work ability maintenance took place at the individual level, but an obvious need for more structural collaboration emerged by several suggestions made by the participants (e.g. better information exchange between the school and occupational health care, improved collaboration between the school staff and the co-operative partners). The progress in collaboration may lead towards spontaneous collaboration, in which case both actors have the same target and work in a planned, continuous and natural way to ensure work ability maintenance in the school community. PMID- 16308171 TI - The Contact Hypothesis: an exploration of its further potential in interprofessional education. AB - This paper highlights the research challenges that face researchers wishing to build the evidence base around interprofessional education (IPE). It concentrates specifically on the short-term impact of IPE on a student population. The Contact Hypothesis is a particularly useful theoretical framework to address these challenges as well as guide the development of IPE interventions. A brief description of this theory and the closely-related theories of social identity and categorization is made in order to support and clarify this theoretical position. The application of the Contact Hypothesis as it has already been made in the IPE field is also described. The paper then addresses how the Contact Hypothesis can be further utilized to address IPE research needs. Through consideration of critique of this theory outside of this field, the development of this framework beyond its early applications to the IPE field are addressed in terms of future direction, the caveats and models of IPE that now require empirical testing. PMID- 16308172 TI - Beginning the process of teamwork: design, implementation and evaluation of an inter-professional education intervention for first year undergraduate students. AB - An evidence-based interprofessional educational (IPE) intervention involving first year undergraduate students studying medicine, nursing, physiotherapy and occupational therapy was piloted at the University of Liverpool. Campbell's phased approach and Complexity Theory guided development of the intervention and its evaluation. The intervention included a staff-training programme, e-learning materials and interprofessional team working skills workshops. A multi method study design was used to evaluate outcomes and the processes by which the outcomes had transpired. The first year cohort of students (n=442) was invited to attend the pilots. Fifty-four per cent (n=237) opted to attend. Findings showed that the intervention promoted theoretical learning about team working. It enabled the students to learn with and from each other (p<0.001), it significantly raised awareness about collaborative practice (p<0.05), and its link to improving the effectiveness of care delivery (p<0.01). The qualitative data showed that it served to increase students' confidence in their own professional identity and helped them to value difference making them better prepared for clinical placement. The findings support the need to start IPE early in students' training before professional doctrines have been built into their learning. As a result of the findings, the intervention has become compulsory for students to attend and the project has evolved to include trained service users/carers as co-facilitators of the workshops. It is also working on strengthening e-learning by integrating the generic materials into the curricula of all courses. Alongside this, strategies are being explored for interprofessional learning in practice. PMID- 16308173 TI - Case-based learning in cross-professional groups - the development of a pre registration interprofessional learning programme. AB - This paper describes the development and evaluation of an interprofessional learning (IPL) programme at the pre-registration level. The principal aim of the study was to investigate whether case-based learning in cross-professional groups is a feasible and an effective way to conduct interprofessional education (IPE). Student volunteers from five different health professional training programmes were allocated to two groups: an intervention group and a control group. Interprofessional attitudes of all students were measured at the beginning and at the end of the study. Group members fed back their views about their learning experience after the 9-week long intervention. The study reports significant effects of the intervention on students' attitudes to different health professions. For example, students in the intervention group tended to view each profession as more 'caring' when compared to the control group. Student feedback was positive, with the main message to integrate the programme in the timetable and to introduce an opportunity for IPE in future years. The initial findings reported in this paper show that this is a feasible and an effective way to deliver IPE across the wide range of professions in the study and that the learning programme was viewed positively by the students who took part. PMID- 16308174 TI - Reducing errors of omission in chronic disease management. PMID- 16308175 TI - Scoping interprofessional education and training in North East London. PMID- 16308176 TI - How do we care for our students? A three-year follow-up of medical, nursing, physiotherapy and occupational therapy students in Sweden. PMID- 16308180 TI - Upper gastrointestinal haemorrhage complicating antiplatelet treatment with aspirin and/or clopidogrel: where we are now? AB - A large number of patients require antiplatelet therapy (mainly aspirin and/or clopidogrel). Recent studies suggest that the combination of these agents is useful in patients with acute coronary syndrome and after percutaneous coronary intervention with stent placement. On the other hand, bleeding complications, most of which arise from the upper gastrointestinal (UGI) tract, can limit the use of antiplatelet drugs. Clopidogrel appears to be associated with fewer UGI side effects and bleeding compared with aspirin. However, a history of previous UGI bleeding is a major risk factor for clopidogrel-associated bleeding. The use of proton-pump inhibitors (PPIs) decreases the rate of UGI bleeding in patients receiving aspirin or clopidogrel. Furthermore, a recent study suggested that the administration of low-dose aspirin plus high-dose esomeprazole (a potent PPI) was associated with fewer episodes of UGI bleeding than clopidogrel alone in patients with a history of recent UGI haemorrhage. However, this study had several limitations and its results should be cautiously extrapolated into clinical practice. The combination of aspirin plus clopidogrel increases the risk of UGI bleeding. Unfortunately, there are no data on the effect of PPI prophylaxis in this setting. Available evidence suggests that where aspirin and/or clopidogrel are to be started or continued in patients with a recent history of UGI ulceration or bleeding (after ulcer healing and eradication of H. pylori infection), treatment with a PPI is a useful precaution. The patients should also be carefully monitored for recurrence of UGI bleeding. PMID- 16308181 TI - A new definition of aspirin non-responsiveness by platelet function analyzer-100 and its predictors. AB - BACKGROUND: Aspirin non-responsiveness has been described as having a normal closure time (CT) by platelet function analyzer (PFA)-100 assay despite confirmed treatment with aspirin. There is no standard definition of aspirin non responsiveness by PFA-100, with a variety of cut-off values having been used. We proposed an alternative definition of aspirin non-responsiveness by PFA-100 assay. METHODS: One hundred eighty-four patients with diagnosis of stable coronary artery disease or diabetes mellitus were included in the study. Blood samples were drawn before and after the 7 days of aspirin therapy. An individual was labelled as aspirin non-responder if his/her post-aspirin CT was not 2SD above his/her baseline CT, where SD was calculated from the baseline CTs of the study population. Aspirin non-responsiveness was also defined as having a normal post-aspirin CT (< or =193 s) regardless of pre-aspirin CT. RESULTS: The baseline CT ranged 82-187 s (mean 129.1 +/- 27.5, median 128 s) in the study population. At the end of 1 week of aspirin administration, CT increased to a mean of 260.7 +/- 63.6 s (range 102-301). According to our definition, 28 (15.2%) of 184 patients were aspirin non-responders. Univariate analysis indicated that aspirin non-responsiveness was closely associated with gender (P = 0.012) diabetes (P = 0.006), smoking (P = 0.0496) and hypertension (P = 0.021). Multivariate analysis identified diabetes (P = 0.016) as the only significant independent predictor for the presence of aspirin non-responsiveness. Thirty-four of 184 patients (18.5%) classified as aspirin non-responders according to the second criteria. Seven patients with prolongation of post-aspirin CT more than 2SD were classified as aspirin non-responders by the second criteria. Only 1 patient without prolongation of CT more than 2SD was classified as aspirin responsive by the second criteria. CONCLUSION: Definition of aspirin non-responsiveness as post aspirin CTs < or =193 s might overestimate the prevalence of aspirin non responsiveness. Nevertheless, definition of aspirin non-responsiveness by PFA-100 must be standardized and its utility as a predictor of cardiovascular events needs to be further investigated. PMID- 16308182 TI - Neutrophil secretory defect in the gray platelet syndrome: a new case. AB - We report the case of a 60-year-old woman who was newly diagnosed for the gray platelet syndrome (GPS). This patient had long-term thrombocytopenia which had been initially misdiagnosed as idiopathic thrombocytopenic purpura (ITP). Blood smear displayed characteristic gray platelets, allowing the diagnosis to be made, which was confirmed by electron microscopy (EM). Polymorphonuclear neutrophils (PMN) appeared poorly granulated on the May-Grunwald-Giemsa-stained blood smear. Flow cytometry analysis of PMN demonstrated increased expression of CD35, CD11b and CD18 at resting PMN surface, without any changes after fMLP stimulation. Ultrastructural study retrieved a decreased number of myeloperoxidase (MPO) negative secondary granules in PMN. Immunolabeling confirmed the presence of membrane proteins and the absence of soluble content in platelet and megakaryocyte (MK) alpha-granules, and the decrease of secondary granules and secretory vesicles in PMN. This new observation demonstrates that the impairment of the secretory compartment of PMN is definitely a hallmark of GPS, and that the detection of these subtle abnormalities should be searched with adequate and up to-date technical approaches. PMID- 16308183 TI - Defective platelet beta-N-acetyl hexosaminidase content and release in chronic myeloproliferative disorders. AB - BACKGROUND AND OBJECTIVES: Abnormalities of platelet function or structure are a hallmark of chronic myeloproliferative disorders (MPD). In vivo platelet activation with the release of alpha- and delta-granules in the circulation is one of the most frequently described alterations in MPD. Platelets contain and release upon activation also lysosomes, and in particular beta-N acetylhexosaminidase (Hex). We have assessed whether the content and in vivo release of Hex of platelets from MPD patients is altered. DESIGN AND METHODS: Twenty-three MPD patients were compared with 19 age- and sex-matched healthy controls. The activity of platelet beta-N-acetylhexosaminidase was measured in plasma, serum and in the capillary blood emerging from the skin wound inflicted for the measurement of the bleeding time. Lysosome integral membrane protein (LIMP or CD63), lysosome-associated membrane protein (LAMP-2 or CD107b) and P selectin were evaluated by flow cytometry. Platelet aggregation in vitro and the release of beta-N-acetylhexosaminidase, ATP and beta-thromboglobulin were performed to study platelet reactivity. RESULTS: Hex levels in plasma were significantly higher in MPD than in controls while the release of Hex in the bleeding time blood, i.e. at a localized site of in vivo platelet plug formation, was lower in MPD and the platelet content of Hex was reduced. These changes were accompanied by in vivo platelet activation. Finally, the isoenzymatic pattern of Hex was altered in platelets of MPD patients, with a reduced amount of the Hex A isoform as compared with controls.b INTERPRETATIONS AND CONCLUSIONS: MPD patients present an altered platelet Hex content and release; prospective studies to assess whether altered platelet Hex is related to thrombotic/hemorrhagic complications and/or tissue fibrosis in MPD are warranted. PMID- 16308184 TI - Does meta-chlorophenylpiperazine (mCPP) activate human platelets? AB - mCPP (meta-chlorophenylpiperazine), an agonist at serotonin (5-hydroxytryptamine, 5-HT) 5-HT2 receptors, has been used as a probe of serotonergic function. We assessed its effect on platelet activation by measuring median platelet volume (MPV), the Sonoclot (SCT) pattern and plasma and intraplatelet serotonin. (a) In vitro study: MPV was measured (n = 7) using a high-resolution channelyzer: Saline (median and range (5.23 fl; 5.10-6.18) vs. mCPP (5.36; 5.10-6.44) P = 0.03; ADP (5.42; 5.29-6.44) vs. ADP + mCPP (5.67; 5.42-6.63) P = 0.02; mCPP (5.36; 5.10 6.44) vs. ADP + mCPP (5.67; 5.42-6.63) P = 0.02. Therefore, mCPP increases the MPV and enhances the effect of ADP. (b) In vivo study: The SCT time to inflection (TI) and time to peak (TP) were measured following the oral administration of mCPP (0.5 mg/kg) or aspirin (300 mg) (n = 10). Ingestion of mCPP significantly shortened TI and TP indicating platelet activation. TI: 0 h (mean +/- SD: 10.2 +/ 2.0 min) vs. 6 h (9.3 +/- 1.5) P = 0.03; TP: 0 h (31.9 +/- 7.6) vs. 6 h (23.1 +/ 2.9) P = 0.01. Aspirin had no effect on TI or TP. There were no significant changes in plasma and intraplatelet 5-HT. It is concluded that mCPP activates human platelets via 5-HT receptors. PMID- 16308185 TI - The influence of bromelain on platelet count and platelet activity in vitro. AB - Bromelain is a general name for a family of sulfhydryl-containing, proteolytic enzymes from the pineapple plant. The aim of the present study was to investigate the influence of bromelain on platelet count, platelet aggregation and platelet activity in vitro. Blood samples were taken from the antecubital vein of 10 healthy male non-smokers. Platelet count decreased after incubation with 2.5 and 5 mg bromelain/ml from 277 +/- 17 platelets/nl before to 256 +/- 21 and 247 +/- 19 platelets/nl after the treatment. The ADP and TRAP-6 induced platelet aggregation led to a significant decrease after the incubation with 2.5 mg (ADP: 48.6 +/- 25.7%; TRAP-6: 49.6 +/- 28.9%) or 5 mg (ADP: 5.0 +/- 4.6%; TRAP-6: 9.0 +/- 4.9%) bromelain/ml in comparison to control (ADP: 81.4 +/- 5.0%; TRAP-6: 77.4 +/- 10.4%). The percentage of unstimulated CD62P positive platelets which were investigated by flow cytometry was minimally higher after incubation with 5 mg bromelain/ml (0.57 +/- 0.48% PC) in comparison to control (0.22 +/- 0.11% PC), but after TRAP-6 stimulation the incubation with 5 mg bromelain/ml led to a remarkable decrease in comparison to the untreated control (50.4 +/- 20.2 to 0.9 +/- 0.8% PC). The changes of CD62P (TRAP-stimulated) and the results of platelet aggregation after incubation with bromelain in vitro may demonstrate the potential of bromelain as a substance for platelet inhibition. PMID- 16308186 TI - The Retention Index Test Homburg (RTH-II): Response to clopidogrel and comparison with aggregometry and CD62P-expression. AB - The Retention Index Test Homburg (RTH-II) is quoted to detect effects of shear stress on platelets, which involve ADP receptor signaling. RTH-II might be a tool for monitoring antiplatelet therapy for compounds that interfere with ADP induced platelet activation and secretion. In a series of investigations, we used an ADP (2 microM) triggered RTH-II in parallel with light-transmittance aggregometry and flow cytometry in subjects before and after clopidogrel. A loading dose of 225 mg clopidogrel leads to a significant reduction (p < 0.01) in the ADP-stimulated retention index (RI) from 69 +/- 15 to 48 +/- 21%, in the aggregation response to 5 microM ADP (from 50 +/- 20 to 29 +/- 21%) and the expression of CD62P (from 64 +/- 11 to 41 +/- 17%). Correlation analysis showed that the RI corresponds significantly to CD62P-expression (p < 0.01) but not to aggregation. We also found a strong correlation (p < 0.01) between the ADP-stimulated RI and the expression of CD62P after stimulation with 2 microM ADP, whereas no correlation was seen for RI vs. binding of PAC-1 or aggregation. Platelets not retained in the filter had lower CD62P expression than measured in the sample before the filter passage (54 vs. 35%). A direct interaction of CD62P with platelet ligands might lead to enhanced retention in RTH and explain the correlation of RI with CD62P expression. The RTH-II might be a simple and easy to handle platelet function assay for monitoring effects on P2Y(12)-inhibitors on platelet degranulation, perhaps in addition to aggregometry. PMID- 16308187 TI - Evaluation of platelet function, a method comparison. AB - Platelet function can be studied using many different methods why it is of interest to understand how data from different assays relate to each other. In the present study we compare two methods suitable for screening purposes with two established although laborious methods, impedance aggregometry and platelet-rich plasma (PRP) aggregation. The alternative assays tested were: (i) exposure of active alphaIIbbeta3, in diluted whole blood and (ii) whole blood aggregation assessed by residual platelet counting. The fibrinogen receptor activation assay was found to have the lower variability, higher sensitivity to ADP, and higher signal to noise ratio compared with residual platelet counting. The sensitivity and response profile of the fibrinogen receptor activation assay and residual platelet counting were more similar to PRP aggregation than to impedance aggregometry, whereas impedance aggregometry displayed lower sensitivity to ADP. The two alternative assays correlated well with PRP aggregation as well as with each other. The fibrinogen receptor activation assay displayed the highest potency for AR-C69931MX, possibly due to a lower protein content compared with residual platelet counting. The two studied assays compare well with the more established assays, and are thus both good alternatives for platelet function testing and evaluation of new potential platelet antagonists. PMID- 16308188 TI - Coated-platelets retain amyloid precursor protein on their surface. AB - Coated-Platelets are a subset of platelets produced by dual-agonist activation with collagen plus thrombin and are characterized by strong retention of several procoagulant, alpha-granule proteins on the cell surface. In this report we demonstrate that coated-platelets also retain full-length amyloid precursor protein (APP) on their surface in contrast to the cleavage of APP in platelets activated with a single agonist. In addition, western blot analysis indicated that APP is derivatized during coated-platelet synthesis. We subsequently measured coated-platelet production in patients with Alzheimer's disease (AD). Twenty-two AD patients showed a wide distribution of coated-platelet values; however the least impaired AD patients produced coated-platelets at a level significantly above that of aged controls (41.0 +/- 9.9 vs. 28.7 +/- 11.4%; mean +/- 1SD; p = 0.017). These findings suggest that coated-platelets may be a model of aberrant APP processing in early AD patients. PMID- 16308189 TI - The formation of platelet-leukocyte aggregates varies during the menstrual cycle. AB - Platelet-leukocyte aggregates are considered to play a significant role in blood coagulation and inflammatory processes. We hypothesized that hormonal changes during the menstrual cycle affect the formation of heterotypic aggregates and therefore may constitute cycle-dependent variations of the susceptibility for thromboembolic events and inflammatory disease. We therefore measured platelet leukocyte interaction by the determination of platelet-leukocyte aggregates (PLA), platelet P-Selectin expression, and platelet fibrinogen receptor activation by PAC-1 binding in 20 healthy women during their menstrual cycle by flow cytometry. The number of platelet-granulocyte aggregates (PGA) and platelet monocyte aggregates (PMA) was higher at ovulation compared to any other time point of the menstrual cycle (p = 0.005, p = 0.022, respectively). Likewise, P Selectin expression peaked on day 14 (p = 0.040). The course of PLA formation during the menstrual cycle followed the course of estrogen levels, strongly suggesting direct effects of estrogen on platelet-leukocyte interaction. The susceptibility to form platelet-leukocyte aggregates that are inducible in vitro by a suboptimal concentration of thrombin receptor activating peptide-6 decreased slightly during the transition from day 1 to 14 (p = 0.040). These data indicate that platelet function varies during particular phases of the normal menstrual cycle. PMID- 16308190 TI - The mean platelet volume in subjects with impaired fasting glucose. AB - Mean platelet volume (MPV), a determinant of platelet function, is a newly emerging risk factor for atherothrombosis. Impaired fasting glucose (IFG) is probably a frequent glycemic disorder in the general population and is considered as a prediabetic state. The present study was designed to evaluate MPV in subjects with IFG compared with diabetic patients and normoglycemic control subjects. We selected 50 patients with type 2 diabetes mellitus, 50 subjects with IFG, and 50 normoglycemic healthy subjects matched for age, gender, and body mass index. MPV was very significantly higher in diabetic and IFG groups than in control group (p < 0.00, p < 0.05, respectively); it was also higher in diabetic group than in IFG group (p < 0.05). Platelet counts were not different among the study groups (p > 0.05). Platelet mass was significantly higher in diabetic and IFG groups than in normotensives (p < 0.00, p < 0.05, respectively); and it was also higher in diabetic group than in IFG group (p < 0.05). MPV and platelet mass were positively correlated with fasting glucose and HbA1c in diabetic and IFG groups (p < 0.05). In conclusion, our data suggests one possible mechanism by which subjects with IFG may be at increased cardiovascular risk. PMID- 16308191 TI - Chemical constituents of the brown alga Dictyopteris divaricata. AB - A novel sesquiterpene-substituted benzoic acid, named dictyvaric acid (1), together with nine known compounds (2-10), have been isolated from the brown alga Dictyopteris divaricata Okam. The structure of 1 was elucidated as 3-[(decahydro 2-hydroxy-2,5,5,8a-tetramethyl-1-naphthalenyl)-methyl]-4-hydroxybenzoic acid by spectroscopic methods, including IR, FABMS, HR-FABMS, 1D and 2D NMR techniques. All compounds were obtained from this species for the first time. PMID- 16308192 TI - A pair of isomeric saponins with cytotoxicity from Albizzia julibrissin. AB - Two new saponins have been isolated from the stem barks of Albizzia julibrissin Durazz, and their structures identified as 3-O-[beta-D-xylopyranosyl-(1 --> 2) beta-D-fucopyranosyl-(1 --> 6)-beta-D-2-deoxy-2-acetoamidoglucopyranosyl]-21-O {(6S)-2-trans-2-hydroxymethyl-6-methyl-6-O-[4-O-((6S)-2-trans-2-hydroxymethyl-6 hydroxy-6-methyl-2,7-octadienoyl)-beta-D-quinovopyranosyl]-2,7-octadienoyl} acacic acid-28-O-beta-D-glucopyranosyl-(1 --> 3)-[alpha-L-arabinofuranosyl-(1 --> 4)]-alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D-glucopyranosyl ester (1) and 3-O [beta-D-xylopyranosyl-(1 --> 2)-beta-D-fucopyranosyl-(1 --> 6)-beta-D-2-deoxy-2 acetoamidoglucopyranosyl]-21-O-{(6S)-2-trans-2-hydroxymethyl-6-methyl-6-O-[3-O ((6S)-2-trans-2-hydroxymethyl-6-hydroxy-6-methyl-2,7-octadienoyl)-beta-D quinovopyranosyl]-2,7-octadienoyl}acacic acid 28-O-beta-D-glucopyranosyl-(1 --> 3)-[alpha-L-arabinofuranosyl-(1 --> 4)]-alpha-L-rhamnopyranosyl-(1 --> 2)-beta-D glucopyranosyl ester (2), based on chemical and spectral evidences, named as julibroside J19 and julibroside J18, respectively. Both compounds show significant inhibition action against HeLa, Bel-7402 and MDA-MB-435 cancer cell lines in vitro. PMID- 16308193 TI - Two new triterpenoid saponins from Symplocos chinensis. AB - Two new triterpenoid saponins, symplocososides X (1) and Y (2) have been isolated from the roots of Symplocos chinensis, and their structures elucidated as 21beta O- cinnamoyl-22alpha-O-(2-methylbutanoyl)-15alpha, 16alpha, 28-trihydroxyolean-12 ene-3beta-O-[beta-D-glucopyranosyl (1 --> 2)]-alpha-L-arabinofuranosyl (1 --> 4) beta-D-glucuronopyranoside (1) and 21beta-O-cinnamoyl-22alpha-O-(2 methylbutanoyl)-15alpha, 16alpha, 28-trihydroxyolean-12-ene-3beta-O-(3-O-acetyl) [beta-D-glucopyranosyl (1 --> 2)]-alpha-L-arabinofuranosyl (1 --> 4)-beta-D glucuronopyranoside (2) by spectral and chemical methods. Their antitumor activities have also been tested. PMID- 16308194 TI - Acretoside, a new sucrose ester from Aristolochia cretica. AB - A new sucrose ester, acretoside, has been isolated from the roots of the Greek endemic species Aristolochia cretica and identified as 6-O-p-coumaroyl-beta-D fructofuranosyl-(2 --> 1)-alpha-D-glucopyranoside (1). In addition, a known sucrose ester, identified as arillatose B, two phenylpropanoid glucose esters, and five derivatives of aristolochic acids have been isolated. Their structures have been elucidated on the basis of MS and NMR data. PMID- 16308195 TI - Novel dimeric alkaloids from the roots of Thalictrum atriplex. AB - Two new bisbenzylisoquinoline alkaloids, neothalfine (1) and thaliatrine (2), together with three known dimeric alkaloids, thalifaberine, thalistine, and thalirecebine, have been isolated from the roots of Thalictrum atriplex Finet et Gagnep. Their structures have been established by spectroscopy. Compound 1 showed in vitro antiplatelet aggregation activities. PMID- 16308196 TI - Two new diterpenoids from Coleus forskohlii. AB - Two new diterpenoids, forskolins I and J, have been isolated in our further investigation on Coleus forskohlii (Willd.) Briq. collected in Yunnan Province. Their structures have been determined as 1alpha,6beta-diacetoxy-7beta,9alpha dihydroxy-8,13-epoxylabd-14-en-11-one (1) and 1alpha,9alpha-dihydroxy-6beta,7beta diacetoxy-8,13-epoxylabd-14-en-11-one (2) by spectral methods (including 1D and 2D NMR techniques). PMID- 16308197 TI - Two new saponins from the bud of Aralia elata (Miq.) Seem. AB - Two new saponins, named congmuyanoside A and congmuyanoside B, have been isolated from the buds of Aralia elata (Miq.) Seem. Their structures have been determined on the basis of chemical and spectroscopic evidence. PMID- 16308198 TI - Isolation and characterization of alpha-(1-->6)-glucans from Cistanche deserticola. AB - Three unique polysaccharides (1-3) have been obtained from the 0.5 M NaOH extract of the stem of Cistanche deserticola Y. C. Ma. The results of methylation analysis, partial acid hydrolysis, 13C, 1H NMR, 1H-1H COSY, HMQC and HMBC spectroscopic analyses indicate that they are all composed of glucose, having a backbone of alpha-(1 --> 6)-glucan, and have different molecular weights. Their structures differ from that of linear starch. PMID- 16308199 TI - Two new triterpenoid saponins from Sarcandra glabra. AB - Two new triterpenoid saponins, named sarcandroside A and B, have been isolated from Sarcandra glabra (Thunb) Nakai. Their structures have been established as 3beta,19alpha,20beta-trihydroxyurs-11,13 (18)-diene-28,20beta-lactone-3-O-beta-D glucopyranosyl (1 --> 3)-[alpha-L-rhamnopyranosyl(1 --> 2)]-beta-D-xylopyranoside (1) and 3-O-beta-D-glucopyranosyl (1 --> 3)-[alpha-L-rhamnopyranosyl(1 --> 2)] beta-D-xylopyranosyl-pomolic acid 28-O-beta-D-glucopyranosyl ester (2) by means of spectral and chemical methods. PMID- 16308200 TI - A new phenanthraquinone from the stems of Bauhinia variegata L. AB - A new phenanthraquinone, named bauhinione (1), has been isolated from Bauhinia variegata L., and its structure has been elucidated as 2,7-dimethoxy-3-methyl 9,10-dihydrophenanthrene-1,4-dione on the basis of spectroscopic analysis. PMID- 16308201 TI - A new dicoumarinoid glycoside from Daphne giraldii. AB - A new dicoumarinoid glycoside, named giraldoid A (1), has been isolated from Daphne giraldii Nitsche. The structure of 1 was determined as 7-O-beta-glucosyl-8 (7-hydroxy-2H-1-benzopyran-2-one-8-)yl-2H-1-benzopyran-2-one on the basis of chemical reactions and spectroscopic methods. PMID- 16308202 TI - Microbial transformation of fraxinellone by Aspergillus niger. AB - Microbial transformation of fraxinellone (1) by Aspergillus niger (AS 3.421) has been carried out. Two converted products, dasycarpol (2) and a new compound fraxinigerllone (3) were obtained. Their structures have been identified on the basis of spectroscopic methods. Dasycarpol shows moderate inhibitory activity on lung cancer cell line A549. PMID- 16308203 TI - Chemical constituents from Schisandra plena. AB - The chemical constituents of the leaves and stems of Schisandra plena are described for the first time. This investigation has resulted in the isolation of a new sesquiterpenoid, plenoxide (1). In addition, eleven known compounds, including sesquiterpenoids, coumarins, flavanones, triterpenoids and steroids have also been isolated. The structure and stereochemistry of 1 has been determined on the basis of spectroscopic analysis. Detailed analysis of 2D NMR data led to the conclusion that the chemical shifts of earlier compounds similar to bullatantriol need revision. PMID- 16308204 TI - A novel sulphur glycoside from the seeds of Descurainia sophia (L.). AB - A new sulphur glycoside, named descurainoside (1), and the known compound sinapic acid (2) have been isolated from the seeds of Descurainia sophia (L.) Webb ex Prantl. The structure of 1 has been identified as (1R,6S,8R,9S,10S)-9,10 dihydroxy-4-[(4-hydroxy-3,5-dimethoxyphenyl)methylene]-8-(hydroxymethyl)-2,7 dioxa-5-thiabicyclo[4.4.0]decan-3-one by means of physico-chemical properties and spectroscopic methods (1D and 2D NMR, HRMS, ESI-MS). PMID- 16308205 TI - A new bisabolane sesquiterpenoid from Euphorbia chrysocoma. AB - The new sesquiterpenoid (6R)-2-chloro-6-[(1S)-1,5-dimethylhex-4-en-1-yl]-3 methylcyclohex-2-en-1-one (1), together with ten known compounds, (6R)-6-[(1S) 1,5-dimethylhex-4-en-1-yl]-3-methylcyclohex-2-en-1-one (2), bauerenol acetate (3), lupenone (4), alpha-amyrenone (5), beta-sitosterol (6), stigmasterol (7), beta-amyrin (8), ursolic acid (9), betulinic acid (10), scopolin (11), have been isolated from the roots of Euphorbia chrysocoma Levl. et Vant. Their structures have been elucidated by spectroscopic data. PMID- 16308206 TI - A new cerebroside from Uvaria tonkinensis var. subglabra. AB - A new cerebroside, subglain A (1), together with five known compounds (2-6) have been isolated from the stems of Uvaria tonkinensis var. subglabra. The structure of 1 has been determined to be 1-O-beta-D-glucopyranosyl-(2S,3S,4R,8Z,2'R)-2-[N (2'-hydroxytetracosanyl)-N-(1'',2''-dihydroxyethyl)-amide]-8-tetradecene-1,3,4 triol by spectroscopic evidence. The known compounds were identified as schisandriside (2), erythritol (3), beta-D-glucopyranose (4), kaempferol-3,7-O alpha-L-dirhamnoside (5), and (+)-lyoniresinol (6). PMID- 16308207 TI - Doxorubicin loaded pH-sensitive micelle targeting acidic extracellular pH of human ovarian A2780 tumor in mice. AB - The purpose of this study was to examine the efficacy of a chemotherapeutic drug, doxorubicin (DOX), loaded in pH-sensitive micelles poly(l-histidine) (M(n):5K)-b PEG (M(n):5K) micelles. The micelles were designed to target the acidic extracellular pH of solid tumors. Studies of pH-dependent cytotoxicity, growth rate of the tumor, pharmacokinetics and biodistribution were conducted. In vitro DOX uptake upon A2780 cells by incubating the cells in a pH 6.8 complete medium at a concentration of 20 microg DOX/ml in the micelle formulation was more than five times that of pH 7.4 condition for initial 20 min. In vivo pharmacokinetic data showed that AUC (area under concentration curve) and half life time (t(1/2)) (plasma half life) of DOX in the pH sensitive micelles increased about 5.8- and 5.2-fold of free DOX in phosphate buffered saline (PBS), respectively. It appeared that DOX in the pH-sensitive micelles preferentially accumulated in the tumor site. The distributions at 12 h post injection in other organs including liver, kidney, spleen, lung and heart were not significantly different from those of DOX in PBS at a 6 mg DOX/kg dose. The in vivo test of anti-tumor activity was performed with human ovarian carcinoma A2780 which was subcutaneously xenografted in female nu/nu athymic mice. The pH-sensitive micelle formulation significantly retarded tumor growth rate without serious body weight loss. The triggered drug release by the reduced tumor pH is believed to be a major mechanism of the observed efficacy after passive accumulation of the micelles by EPR effect. This may have resulted in a local high dose of drug in the tested solid tumor. PMID- 16308208 TI - Generation of immunoliposomes using recombinant single-chain Fv fragments bound to Ni-NTA-liposomes. AB - Recombinant single-chain Fv antibody fragments (scFv) can be combined with liposomes to generate immunoliposomes for targeted drug delivery. Recent studies have shown that scFv molecules modified to express a C-terminal cysteine residue can be used for site-directed chemical conjugation. Here, we present a new method by immobilizing scFv fragments via their C-terminal hexahistidyl-tag on liposomes containing Ni-NTA-lipids (Ni-NTA-DOGS) in their lipid bilayer without the need to introduce additional reactive groups in the protein. Using an anti-endoglin scFv as a model antibody, we could show that scFv molecules are efficiently immobilized on the liposome surface and that these immunoliposomes bind specifically and strongly to endoglin-expressing endothelial cells. This approach allows for a rapid and flexible generation of target cell-specific immunoliposomes. PMID- 16308209 TI - Aortic drug delivery of dexamethasone palmitate incorporated into lipid microspheres and its antiatherosclerotic effect in atherogenic mice. AB - In order to confirm the efficacy of dexamethasone (DXM) palmitate incorporated into lipid microspheres (d-lipo) on atherosclerosis, the aortic drug delivery by d-lipo and its antiatherosclerotic effect were investigated. In an in vitro uptake experiment, d-lipo or DXM was added to macrophages and foam cells, and then incubated for 1, 4, 8 and 24 h at 37 degrees C. The uptake of drug by these cells after addition of d-lipo was higher than that of DXM at each time point. In an in vitro pharmacological experiment, the macrophages and foam cells were incubated with d-lipo or DXM for 24 h at 37 degrees C. The inhibitory effect of d lipo on cellular cholesterol ester (CE) accumulation in these cells was significantly more potent than that of DXM. In an in vivo pharmacokinetic experiment, d-lipo or DXM was intravenously administered to atherogenic mice, and then aorta was collected at 1, 8, and 24 h after administration. The aortic drug concentration after administration of d-lipo to atherogenic mice was higher than that of DXM at each time point. In an in vivo pharmacological experiment, d-lipo or DXM was intravenously administered to atherogenic mice once a week for 7 weeks. The inhibitory effect of d-lipo on the aortic CE accumulation in atherogenic mice was significantly more potent than that of DXM. These findings suggest that efficient drug delivery to the atherosclerotic lesions by d-lipo produces an excellent antiatherosclerotic effect at a lower dose. Therefore, d lipo may be useful for the development of drug delivery systems for atherosclerotic therapy. PMID- 16308210 TI - Cutaneous DNA delivery and gene expression in ex vivo human skin explants via wet etch micro-fabricated micro-needles. AB - Micro-needle arrays increase skin permeability by forming channels through the outer physical barrier, without stimulating pain receptors populating the underlying dermis. It was postulated that micro-needle arrays could facilitate transfer of DNA to human skin epidermis for cutaneous gene therapy applications. Platinum-coated "wet-etch" silicon micro-needles were shown to be of appropriate dimensions to create micro-conduits, approximately 50 microm in diameter, extending through the stratum corneum (SC) and viable epidermis. Following optimisation of skin explant culturing techniques and confirmation of tissue viability, the ability of the micro-needles to mediate gene expression was demonstrated using the beta-galactosidase reporter gene. Preliminary studies confirmed localised delivery, cellular internalisation and subsequent gene expression of pDNA following micro-needle disruption of skin. A combination of this innovative gene delivery platform and the ex vivo skin culture model will be further exploited to optimise cutaneous DNA delivery and address fundamental questions regarding gene expression in skin. PMID- 16308211 TI - Immunomodulator tuftsin increases the susceptibility of Cryptococcus neoformans to liposomal amphotericin B in immunocompetent BALB/c mice. AB - The co-administration of immunomodulators and antibiotics has been proved very successful for treatment of opportunistic infectious diseases. In the present study, we evaluated the combination of liposomal amphotericin B (lip-Amp B) and immunomodulator tuftsin to cure Cryptococcus neoformans infection in BALB/c mice. Mice infected with C. neoformans were treated with Amp B deoxycholate and tuftsin free or tuftsin-loaded Amp B liposomes. The results of the present study demonstrated higher efficacy of tuftsin-loaded Amp B liposomes against experimental murine cryptococcosis, in terms of enhanced survival rate and reduced fungal burden in organs (lungs and brain) of the treated mice. Interestingly, pre-treatment of mice with liposomal tuftsin before challenging them with the C. neoformans infection resulted in 100% survival of the treated animals followed by treatment with lip-Amp B. Immunomodulator-based therapy seems likely to be more beneficial for treatment of fungal infectious diseases. PMID- 16308212 TI - Polypropylenimine dendrimer-induced gene expression changes: the effect of complexation with DNA, dendrimer generation and cell type. AB - Polypropylenimine (PPI) dendrimers appear attractive non-viral vectors for the delivery of genes, antisense oligonucleotides, and small interfering RNA (siRNA). However, the effects of these synthetic gene delivery vectors on global gene expression are poorly understood. Here we have examined the toxicogenomics of generation 2 (DAB-8) and generation 3 (DAB-16) PPI dendrimers in two human cell lines. At concentrations and treatment protocols routinely used for gene and oligonucleotide transfection, PPI dendrimers alone elicited marked changes in endogenous gene expression in A431 epithelial cells. The extent of PPI-induced gene changes appeared to be dependent on the dendrimer generation as the number of genes affected was greater with G3 compared to G2 PPI dendrimers in A431 cells. The signature of DAB16-induced gene changes in A549 cells was different to those elicited in A431 cells implying a strong dependence on cell type. The DAB 16 polymer complexed with DNA (dendriplexes) also elicited marked gene expression changes in A549 cells but with a signature that was different from the polymer alone implying that dendriplexes are "recognised" by cells as chemical entities that are distinct from the polymer alone. Alterations in expression of a variety of gene ontologies were observed including those involved in defence responses, cell proliferation and apoptosis. Although there was a tendency for increased DNA damage in cells treated with DAB16 alone or its DNA dendriplexes as detected by the COMET assay, these differences were not statistically significant. These data show for the first time that PPI-dendrimers, separate from their capability as transfection reagents, can intrinsically alter the expression of many endogenous genes that could potentially lead to them exerting multiple biological effects in cells. The impact and consequences of polymer-induced gene changes should guide their rational use as delivery systems for gene-based therapeutics. PMID- 16308213 TI - Aetiology of community-acquired pneumonia: serological results of a paediatric survey. AB - Serological methods are routinely used in the diagnosis of viral and atypical bacterial respiratory infections. Recently, they have also been applied to typical bacteria, such as Streptococcus pneumoniae. The aim of this study was to determine the aetiology of paediatric community-acquired pneumonia (CAP) in both ambulatory and hospitalized patients, by using antibody assays. During a 15-month prospective surveillance, paired sera were studied for antibodies to 14 microbes in 101 children with symptoms of acute infection and infiltrates compatible with pneumonia on chest radiographs. A potential causative agent was detected in 66 (65%) patients. Evidence of bacterial, viral and mixed viral-bacterial infection was demonstrated in 44%, 42% and 20% of the CAP cases, respectively. The most commonly found agents included Mycoplasma pneumoniae (27%), Pneumococcus (18%) and respiratory syncytial virus (17%). Human metapneumovirus (hMPV) was detected in 5 (5%) children. Pneumococcal infections were evenly distributed among the age groups studied. Our results confirm the role of S. pneumoniae in paediatric CAP at all ages, those of M. pneumoniae at >2 y of age and emphasize the emerging role of hMPV. The high proportion of mixed viral-bacterial infections highlights the need to treat all children with CAP with antibiotics. PMID- 16308214 TI - Respiratory diphtheria among highly vaccinated military trainees in Latvia: improved protection from DT compared with Td booster vaccination. AB - An outbreak of respiratory diphtheria occurred among highly-vaccinated trainees at a Latvian military academy in August-September 2000. We reviewed immunization, clinical and laboratory records and administered a questionnaire to obtain data on exposure factors. Among 207 trainees, 45 (22%) diphtheria cases and 79 (38%) carriers of toxigenic Corynebacterium diphtheriae were identified. All patients survived; 1 had severe myocarditis. Sharing cups was a risk factor for infection. Over 85% of trainees had received > or =5 doses of diphtheria toxoid. Neither infection nor disease was associated with the number of doses or interval since last dose. However, the risk of disease was lower and diphtheria antitoxin levels were higher among trainees who received their last booster dose with higher antigen diphtheria toxoid (DT) instead of lower-antigen Td. Outbreaks of mild diphtheria can occur among highly-vaccinated persons living in crowded conditions with intense exposure; high-antigen diphtheria booster-vaccination might provide better protection under these conditions. PMID- 16308215 TI - Economic evaluation of pneumococcal conjugate vaccination in Finland. AB - The aim of this study was to evaluate cost-effectiveness of pneumococcal conjugate vaccine (PCV7) in children <5 y of age. A Markov simulation model was used to compare the cost-effectiveness of 4 doses (assumed 50.5 euros per dose) of PCV7 with no intervention. Only direct effects of the vaccine were taken into account. In Finland, vaccination of a birth cohort of 57,500 healthy infants would potentially prevent annually 60 cases of invasive PD, 1,400 cases of pneumococcal pneumonia, 15,000 episodes of acute otitis media, 3,000 otological surgery procedures and 0.9 deaths in children aged <5 y. Investing 12.0 million euros to vaccinate a birth cohort would save annually 6.3 million euros in medical, and 2.0 million euros in productivity and other, costs. Therefore, investing 1 euros in a vaccination programme would return 0.53 euros in medical costs and 0.70 euros in societal costs. In the base case, vaccination would cost society 139,986 euros per life y gained. To achieve cost savings from a health care provider (societal) perspective, without considering herd effects or replacement phenomenon, the price of PCV7 should be 50% (70%) of the price used in the base case. PMID- 16308216 TI - Clinical use of a diagnostic PCR for Francisella tularensis in patients with suspected ulceroglandular tularaemia. AB - A retrospective analysis to evaluate the clinical use of a diagnostic PCR for Francisella tularensis in patients with suspected ulceroglandular tularaemia was performed. 154 samples, 129 from patients with definitive tularaemia and 25 from patients where tularaemia could be ruled out, were analysed. The diagnostic PCR had a specificity of 96%, a sensitivity of 78.3%, and a Positive Predictive Value of 99%. Especially samples from encrusted lesions, even up to 4 weeks old, in patients with tularaemia, were PCR positive to a high degree when taken properly. The diagnostic PCR is useful in suspected ulceroglandular tularaemia, giving a fast and accurate diagnosis. PMID- 16308217 TI - Emergence of Mycobacterium simiae in respiratory specimens. AB - Temporal changes in epidemiology of non-tuberculous mycobacteria (NTM) infections have hardly been explored. Frequency of isolation of different NTM species varies geographically. We aimed to determine whether the distribution of Mycobacteria species isolated from respiratory specimens had changed from 1996 to 2003. We analysed data on 484 patient mycobacterial isolates from 23,483 respiratory specimens submitted to the Laboratory of Microbiology in a tertiary medical centre. The proportion of patients with Mycobacterium tuberculosis isolates decreased from 44.6% in period I (1996-1999) to 20.6% in period II (2000-2003). Among the NTM, the proportion of some species increased significantly (Mycobacterium simiae: 8.4% to 31.6%; Mycobacterium fortuitum 12% to 20%; and Mycobacterium chelonae 4.8% to 11.3%), while others decreased (Mycobacterium avium complex 31.3% to 17.3%; Mycobacterium kansasii 28.9% to 7.5%; and Mycobacterium haemophilum 1.2% to 0%). These findings disclose major temporal changes in the distribution of mycobacterial species in respiratory specimens with an impressive emergence and takeover of M. simiae. PMID- 16308218 TI - Long-term follow-up of children with vesicoureteral reflux with and without antibiotic prophylaxis. AB - The aim of the present study was to obtain data on the outcome of children with persistent vesicoureteral reflux (VUR) after cessation of antibiotic prophylaxis. Children with VUR who had been on antibiotic prophylaxis for at least 2 y and were free of urinary tract infections (UTI), had normal voiding patterns, and no hydronephrosis or new kidney scarring, had antibiotic prophylaxis discontinued, were followed up prospectively with urine cultures, voiding cystourethrography, and technecium-99m dimercaptosuccinate renal scintigraphy. The findings were compared with those of the same patients while they were on antibiotic prophylaxis. In 54 children (39 girls and 15 boys), antibiotic prophylaxis was discontinued. The mean follow-up time on and off antibiotic prophylaxis was 4.4+/ 2.1 and 4.4+/-2.2 y, respectively. Nine UTI episodes occurred during the on- and 8 during the off-prophylaxis period. In 80 of 96 and in 68 of 74 ureters the reflux resolved or downgraded during the on- and off-prophylaxis periods, respectively. No new scar lesions were detected in any of the children. In conclusion, in children with persistent VUR and certain characteristics, antibiotic prophylaxis can be safely discontinued. PMID- 16308219 TI - Drug-resistant tuberculosis in Yangon, Myanmar. AB - The extent of drug resistant tuberculosis (TB) in the capital city of Myanmar, Yangon has not yet been reported. This study aimed to determine the proportion and pattern of drug resistance to first-line anti-TB drugs, among Mycobacterium tuberculosis complex isolates from sputum smear positive TB patients who attended National TB Programme Yangon centres in April-August and October-December 2002. Drug susceptibility was determined by the Mycobacteria Growth Indicator Tube manual system (Becton Dickinson, MD, USA). Of the 567 patients, sputum specimens from 447 (79%) had a positive culture. Of these, 357 isolates (80%) had a susceptibility test result. Isolates from 76 of 259 (29.3%) new patients and from 45 of 98 (45.9%) previously treated patients were resistant to at least 1 of the anti-TB drugs. Resistance to isoniazid (INH) (22.0% vs 40.8%: new vs previously treated patients) and to > or =2 drugs (17.8% vs 29.6%: new vs previously treated patients) was common. Multidrug- resistant TB (MDR-TB) among new and previously treated patients was 4.2% and 18.4%, respectively. INH-resistant (adjusted OR: 2.0, 95% CI 1.1-3.6) and MDR-TB (adjusted OR: 3.4, 95% CI 1.4-8.3) cases were more likely to have taken anti-TB drugs > or =1 month previously. Collectively, prevalence of MDR-TB and TB resistance to > or =2 drugs are not rare in Yangon. PMID- 16308220 TI - Non-typhi Salmonella empyema: case report and review of the literature. AB - A case of non-typhi Salmonella empyema is reported. A review of the literature over the past century found only 13 other empyemas due to Salmonella typhimurium and 25 cases of other non-typhi Salmonella. A review of pleuropulmonary disease due to non-typhi Salmonella is provided along with management recommendations. PMID- 16308221 TI - The dynamics of pulmonary tuberculosis in Colima, Mexico (1999-2002). AB - Tuberculosis is a public health problem in Mexico. From 1999 to 2002, we assessed retrospectively the epidemiological, clinical, and treatment characteristics of pulmonary tuberculosis in the hospitals of the Mexican Institute of Public Health in the state of Colima (Mexico). We included 184 cases diagnosed with pulmonary tuberculosis. A database containing demographic, epidemiological, and clinical information was constructed and analyzed. We estimate a median patient delay of 83 d and a mean treatment delay of 2.3 d. Of 14 cases suspected for multiresistance and microbiologically assayed, 5 were found to carry a multi-drug resistant strain. We also found a significant association between a short patient delay and the presence of hemoptysis (p = 0.002) or dyspnea (p<0.001). 86 patients (46.8%) were sputum smear microscopy negative at the end of treatment and 40 (21.7%) completed treatment giving an overall success rate of 68.5%, which compares unfavorably with the World Health Organization target success rate of 85%. Five (2.7%) patients failed treatment, 10 (5.4%) died, 39 (21.2%) interrupted treatment, and 4 (2.2%) transferred to another reporting unit. A 2002 strategic change in drug distribution seemed to prove successful. PMID- 16308222 TI - Clinical characteristics of nosocomial and community-acquired extraintestinal infections caused by Hafnia alvei. AB - 36 episodes (25 nosocomial and 11 community-acquired) of infections caused by Hafnia alvei showed that this bacterium is responsible for serious infections in adults, specially in hospitalized patients with underlying chronic diseases, subjected to invasive procedures or even under antibiotic treatment. PMID- 16308223 TI - High prevalence of HPV among female students in Finland. AB - We studied the prevalence of HPV DNA among university students within the Helsinki metropolitan area in Finland. First-y students attending a general health examination as well as students visiting a general practitioner for contraception were enrolled. Vaginal self-samples or cervicovaginal swabs were collected from a total of 1469 students. HPV DNA was determined using a liquid hybridization test. Of all students 33.0% were positive for HPV DNA. Of all positive samples 84.3% were positive for high-risk HPV. The study demonstrated a strikingly high HPV DNA prevalence among the young women. Self-collected samples proved to be acceptable for HPV DNA detection using liquid hybridization. PMID- 16308224 TI - Temporarily controlled HIV-1 replication after intravenous immunoglobulin treatment of Guillain-Barre syndrome. AB - HIV establishes a latent infection in resting CD4(+) T-lymphocytes. A possible strategy to eliminate cellular reservoirs in long-lived, HIV-1-infected quiescent CD4(+) T-lymphocytes might be to add T-cell-activating agents to potent antiretroviral therapy. In this report we describe a patient with Guillain-Barre syndrome treated with high dose intravenous immunoglobulin (IVIG) in addition to antiretroviral therapy. A transiently increased viral load and immunoactivation during the IVIG treatment suggest activation of latently infected cells and increased turnover rate of the latent viral reservoir. HIV replication was controlled with plasma viral load <20 copies/ml, for at least 3 months after antiretroviral treatment interruption. CSF neural markers reflecting degenerative processes in the brain during the symptomatic period and follow-up were also analysed. Very high CSF sulfatide concentrations were found indicating that the pathology involves severe demyelination.We hypothesize that IVIG in this case contributed to an activation of latently infected cells, which led to a transient increase in plasma HIV-1 RNA during the IVIG treatment and a long period of undetectable viral load after antiretroviral treatment interruption. Further, this is the first time, to our knowledge, that detailed CSF findings are described in HIV-1 associated GBS. PMID- 16308225 TI - Cell surface expression of FcgammaRI (CD64) on neutrophils and monocytes in patients with influenza A, with and without complications. AB - The expression of the Fcgamma-receptor I (FcgammaRI), CD64 on normal neutrophils is up-regulated during bacterial infections. CD64 is a promising diagnostic tool in the diagnosis of acute infections. The aim was to study surface expressions of CD64 on neutrophils and monocytes in patients with influenza A with and without complications and evaluate these as diagnostic tools in comparison with serum levels of HNL (human neutrophil lipocalin). CD64 expression on neutrophils and monocytes was evaluated by flow cytometry. HNL was assayed by a specific radioimmunoassay. 22 patients with influenza A with or without complications were included and the results compared with those of 29 patients with acute bacterial infections and 29 healthy subjects. Neutrophil expression of CD64 was increased in influenza A with raised proportion expressing CD64 in complicated compared to uncomplicated influenza. The expression was significantly higher in bacterial infections compared to both influenza groups. Serum levels of HNL were raised in all infection groups, but significantly more so in the group with bacterial infection. ROC-curve analysis showed that neutrophil expression of CD64 and the serum levels of HNL had similar diagnostic power in the discrimination between acute bacterial infections and influenza A. Monocyte expression of CD64 was raised in all infections with no differences between subgroups. We conclude that neutrophil expression of CD64 and serum levels of HNL are both promising assays in the distinction between infections caused by bacteria or influenza A, whereas CD64 could identify patients with complications of their influenza A infection. PMID- 16308226 TI - Genetic evolution of HIV in patients remaining on a stable HAART regimen despite insufficient viral suppression. AB - Our objective was to investigate whether steadily increasing resistance levels are inevitable in the course of a failing but unchanged Highly Active Antiretroviral Therapy (HAART) regimen. Patients having an unchanged HAART regimen and a good CD4 response (100 cells/microl above nadir) despite consistent HIV-RNA levels above 200 copies/ml were included in the study. The study period spanned at least 12 months and included 47 plasma samples from 17 patients that were sequenced and analysed with respect to evolutionary changes. At inclusion, the median CD4 count was 300 cells/ml (inter-quartile range (IQR): 231-380) and the median HIV-RNA was 2000 copies/ml (IQR: 1301-6090). Reverse transcription inhibitor (RTI) mutations increased 0.5 mutations per y (STD = 0.8 mutations per y), while major protease inhibitor (PI) resistance mutations increased at a rate of 0.2 mutations per y (STD = 0.8 mutations per y) and minor PI resistance mutations increased at a rate of 0.3 mutations per y (STD = 0.7 mutations per y). The rate at which RTI mutations accumulated decreased during the study period (p = 0.035). Interestingly, the rate of mutation accumulation was not associated with HIV-RNA level. The majority of patients kept accumulating new resistance mutations. However, 3 out of 17 patients with viral failure were caught in an apparent mutational deadlock, thus the development of additional resistance during a failing HAART is not inevitable. We hypothesize that certain patterns of mutations can cause a mutational deadlock where the evolutionary benefit of further resistance mutation is limited if the patient is kept on a stable HAART regimen. PMID- 16308227 TI - Community-acquired pneumonia and bacteraemia in a healthy young woman caused by methicillin-resistant Staphylococcus aureus (MRSA) carrying the genes encoding Panton-Valentine leukocidin (PVL). PMID- 16308228 TI - Splenic abscess caused by Klebsiella pneumoniae and non-Klebsiella pneumoniae in Taiwan: emphasizing risk factors for acquisition of Klebsiella pneumoniae splenic abscess. AB - Klebsiella pneumoniae was recently reported to be the major pathogen causing pyogenic splenic abscess in Taiwan. To better understand the characteristics of K. pneumoniae splenic abscess, which may be helpful in alerting clinicians to this infection entity when dealing with a suspicious patient, patients hospitalized between January 1981 and December 2002 with the diagnosis of splenic abscess were included in a retrospective study. Among the 38 enrolled patients, 9 (23.7%) suffered from K. pneumoniae splenic abscess. Compared to those with non K. pneumoniae splenic abscess, patients suffering from splenic abscess caused by K. pneumoniae had a higher prevalence of underlying diabetes mellitus (88.9% vs 37.9%; p = 0.006) and higher incidence of concomitant liver abscess (44.4% vs 0%; p < 0.001) caused by the same pathogen. When dealing with patients suffering from K. pneumoniae splenic abscess, clinicians should work up to exclude a concomitant liver abscess caused by the same pathogen. PMID- 16308229 TI - Antibiotic drug advertising in medical journals. AB - Advertising is a leading strategy for drug promotion. We analysed 779 advertisements in 24 medical journals, 25% of which featured antibiotics. Antibiotic advertisements showed differences compared to those of other drugs. None addressed the issue of antibiotic resistance. Efforts to prevent antibiotic resistance should take antibiotic advertising into consideration. PMID- 16308230 TI - Measles and rubella vaccination of two million Iranians: complications in vaccinees aged 5-25 years. AB - In 2003 and 2004, 2,049,170 persons aged 5-25 y were vaccinated against measles and rubella in Tehran. Adverse events were actively monitored. A total of 688 adverse events was reported in 476 vaccine recipients with a female dominance (female to male ratio 2.2). Severe adverse events were noted in 13 cases: seizures within 48 h of vaccination (n = 9), encephalopathy (n = 2), flaccid paralysis (n = 1) and anaphylaxis (n = 1). PMID- 16308231 TI - Report of an atypical case of leishmaniasis presented as acute tonsillitis in an immunocompetent patient. AB - Visceral leishmaniasis typically presents with symptoms such as fever, enlargement of the spleen and the liver, hypergammaglobulinaemia and infection of the bone marrow resulting in anaemia and leukopenia. The disease is sporadic in the countries of the Mediterranean basin. We report an unusual case of acute tonsillitis due to tonsillar leishmaniasis, in an immunocompetent 34-y-old male patient. Diagnosis was confirmed by serological tests and histopathological examination following biopsy of the right tonsil. The patient was successfully treated with liposomal amphotericin-B. PMID- 16308232 TI - Early prosthetic valve endocarditis due to Acinetobacter baumannii: a case report and brief review of the literature. AB - Acinetobacter is a highly resistant microorganism, commonly isolated in intensive and post-operative care units. Although rarely reported, it may constitute 1 of the several causes of early prosthetic valve endocarditis. A diffuse, red maculopapular rash may be encountered in patients with Acinetobacter endocarditis. Here we present a case of early prosthetic valve endocarditis due to Acinetobacter baumannii and accompanied by a cutaneous eruption. PMID- 16308233 TI - Fatal non-typeable Haemophilus influenzae sepsis complicated with acute respiratory distress syndrome: case report and literature review. AB - A previously healthy 31-month-old male child became acutely ill with dyspnea and high fever 48 h after admission for acute bronchitis. He experienced sepsis and acute respiratory distress syndrome throughout the subsequent hospitalization, eventually expiring despite aggressive treatment with antibiotics and extracorporeal membrane oxygenation. Blood cultures yielded ampicillin-resistant non-typeable Haemophilus influenzae. To the best of our knowledge, this is the first reported case of fatal non-typeable H. influenzae sepsis and ARDS in a child without an underlying predisposing condition. PMID- 16308234 TI - Central venous catheter-associated fungemia secondary to mucormycosis. AB - Mucormycosis can cause fatal infections in immuno-compromised persons. Rhinocerebral and pulmonary disease predominate. Diagnosis is challenging, dissemination frequent, and mortality high. Positive blood cultures are rare. We report the first case of central venous catheter-associated mucormycosis fungemia (Mucor circinelloides). Early diagnosis and multi-modal therapy led to clinical and microbiological cure. PMID- 16308235 TI - Acute calculous cholecystitis associated with brucellosis: a report of two cases and review of the literature. AB - Acute cholecystitis is a very rare complication of Brucella infections. We report 2 cases of acute cholecystitis due to Brucella and review previously reported cases. PMID- 16308236 TI - A case of Plasmodium falciparum malaria with an exceptionally long incubation period. AB - Clinical symptoms of Plasmodium falciparum malaria normally appear within 2 months of transmission. The incubation period is important for deciding whether a febrile illness is associated with a previous stay in a malarial region. This case report shows that P. falciparum malaria can have a prolonged incubation period. PMID- 16308237 TI - Community acquired pneumonia (CAP) caused by Cryptococcus neoformans in a healthy individual. AB - A 41-y-old male had been diagnosed as having community acquired pneumonia (CAP) with consolidations in the chest radiograph, fever and cough. Since clarithromycin and ss-lactam agents were not effective, bronchoscopic examination was performed. Indian ink staining of bronchial wash smears revealed yeast-like cells with a thick capsule, and Cryptococcus neoformans was isolated several d later. Serum glucuronoxylomannan antigen was > or = x 1024. The patient was treated with itraconazole for 16 weeks. PMID- 16308238 TI - Subclinical hypothyroidism in a patient affected by advanced AIDS and visceral leishmaniasis. AB - Hypothyroidism has been shown to occur in HIV disease. Thyroid function of patients affected by AIDS and leishmaniasis is unknown. Here we report the case of an AIDS advanced patient developing hypothyroidism during leishmaniasis. The thyroid disorder might have been caused by infiltration of the gland by Leishmania. An additive impact of HIV in thyroid function impairment is suggested. PMID- 16308239 TI - Pseudomembranous tracheobronchitis caused by Aspergillus in immunocompromised patients. AB - We report 2 cases of Aspergillus pseudomembranous tracheobronchitis in patients with diabetes. The first patient succumbed to progressive obstructive respiratory failure despite mechanical ventilation and antifungal therapy. However, the second patient survived. Aspergillus tracheobronchitis should be considered in immuno-compromised patients presenting with cough, chest pain, fever, dyspnea and upper airway obstruction. Early bronchoscopy and histologic examination should be performed. Early, appropriate treatment may be life saving. PMID- 16308240 TI - Eristalis tenax as a cause of urinary myiasis. AB - Eristalis tenax, belonging to order Diptera, family Syrphidae seldomly causes intestinal myiasis. Urinary myiasis caused by Eristalis tenax larvae is a rare manifestation found in both humans and other vertebrate animals. We report a 58-y old woman presented with painful mixing and bilateral costo-lumbar pain. The larva in her urine sample was identified as Eristalis tenax related to its typical morphology. PMID- 16308241 TI - Failure of interferon alpha-2b in a patient with West Nile virus meningoencephalitis and acute flaccid paralysis. AB - WNV infection can cause meningoencephalitis (MNE) and acute flaccid paralysis (AFP). Both syndromes are associated with high morbidity and mortality. Interferon alpha-2b (IFN-alpha-2b) inhibits WNV replication in vitro. To date, 5 patients with WNV-CNS disease have recovered neurologic function with IFN-alpha 2b. We report the first failure of IFN-alpha-2b in the treatment of WNV-MNE-AFP. PMID- 16308242 TI - Treatment failure of methicillin-resistant Staphylococcus aureus endocarditis with linezolid. AB - We report a case of methicillin-resistant Staphylococcus aureus endocarditis treated by vancomycin and cotrimoxazole switched to oral linezolid alone with a complete resolution of the vegetation. Two months after discontinuation of treatment, the patient presented a relapse confirmed by pulsed-field gel electrophoresis involving the same linezolid-susceptible strain and rapidly died. PMID- 16308243 TI - Q fever as a cause of pure sensory polyneuropathy -- the six-year itch: a follow up of an indigenous Swedish case. AB - Q fever is common worldwide but extremely rare as a cause of clinical disease in Sweden. Complications such as meningitis and Guillain-Barre syndrome are not uncommon. Chronic polyneuropathy caused by Q fever has until now never been reported in the English-speaking medical community. The first truly indigenous case of clinical Q fever in Sweden has previously been reported by us in this journal. The same patient also developed a pure sensory polyneuropathy. PMID- 16308244 TI - Pseudomonas mendocina spondylodiscitis: a case report and literature review. AB - Pseudomonas mendocina is rarely recovered as a human pathogen. Only 2 human cases have been reported in the literature. Here, we report the third human case and possibly the first 1 to involve spinal infection caused by such an unusual bacterium. PMID- 16308245 TI - Surgical and medical management of rare echinococcosis of the extremities: pre- and post-operative long-term chemotherapy. AB - Skeletal hydatid disease is a rare parasitic infection caused by the tapeworm Echinococcus granulosus. We present the case of a 30-y-old patient with an echinococcosis of the right leg, who was treated with a preoperative albendazole chemotherapy (4 weeks), followed by radical surgical resection and postoperative albendazole treatment (1 y). PMID- 16308246 TI - Infective endocarditis in diabetic patients. PMID- 16308247 TI - Cost of tracheoesophageal prostheses in developing countries. Facing the problem from an internal perspective. AB - CONCLUSIONS: All the discussants of our questionnaire agreed that the price of tracheoesophageal (TE) prostheses was too expensive for developing countries. The problem could be addressed in terms of international laws regarding companies' patent rights. TE prosthesis manufacturers from rich countries could move their manufacturing in part to developing countries. High production standards could allow TE prostheses to be exported to developed countries. Another approach to the problem may be based on a partnership between non-profit-making Western laryngological societies with specific medical and technical prosthetic know-how and local manufacturers. The aim of this cooperation could be the low-cost production of advanced TE prostheses in the developing countries. OBJECTIVES: In communities in the developing world, most laryngeal and hypopharyngeal carcinomas are diagnosed at advanced stages and require total laryngectomy. Prosthetic TE voice restoration is the method of choice for voice rehabilitation after total laryngectomy in developed countries. Unacceptably high costs are a significant limitation to Third World use of TE voice prostheses. The aims of this paper are to discuss the consequences of the high costs of TE prostheses in developing countries with head and neck surgeons working in Third World hospitals and to propose how European and American laryngological societies can promote TE speech in the developing countries. MATERIAL AND METHODS: A questionnaire was given to a group of expert head and neck surgeons working in developing countries and their answers and suggestions discussed. PMID- 16308248 TI - Estrogen and hearing: a summary of recent investigations. AB - Is the female sex steroid estrogen the key to preserved hearing in the aging human? This question remains unanswered, but hearing loss is more profound in elderly males than females. There are also well-known sex differences in the auditory brainstem response (ABR), i.e. women have shorter latencies than men. Moreover, menopausal women who are administered hormone replacement therapy have slightly better hearing than those who are not, and women with Turner's syndrome (45,X), who are biologically estrogen-deficient, show longer ABR latencies and early presbyacusis. These findings are also supported by animal experiments. When boosted with estrogen or testosterone the non-reproductive female midshipman fish alters its inner ear auditory mechanism so that it can hear the male's hum-like call. If estrogen receptor beta is knocked out in mice, severe progressive hearing loss occurs, leading to early deafness. In apparent contradiction to these findings, there have been case reports suggesting that hormone replacement therapy and oral contraceptive use can lead to hearing loss, but of another type, namely acute sudden deafness. Such contradictory aspects of the action of estrogen are commonly found and may spring from the fact that there are two estrogen receptors, alpha and beta, both of which are present in the inner ear of mice, rats and humans. Knowing how sex steroids can alter hearing ability may give important clues as to how estrogen can preserve hearing in humans. In this review we present a summary of current knowledge about hearing and estrogen. PMID- 16308249 TI - Protective effect of edaravone against the ototoxicity of Pseudomonas aeruginosa exotoxin A. AB - CONCLUSION: Our findings suggest that edaravone can protect against cochlear damage caused by Pseudomonas aeruginosa exotoxin A (PaExoA). OBJECTIVE: To analyze the protective effect of a free radical scavenger, edaravone, against the ototoxicity resulting from exposure of the middle ear to PaExoA. MATERIAL AND METHODS: In nine groups of albino rats the following solutions were instilled either via the tympanic membrane into the round window niche [intratympanically (i.t.)] or intravenously (i.v.): edaravone (i.v.); edaravone (i.t.); PaExoA (i.t.) + edaravone (i.t.; simultaneously); PaExoA (i.t.) + edaravone (i.t.; 1 h after); PaExoA (i.t.) + edaravone (i.t.; 24 h after); PaExoA (i.t.) + edaravone (i.v.; simultaneously); PaExoA (i.t.) + edaravone (i.v.; 1 h after); PaExoA (i.t.) + edaravone (i.v.; 24 h after); PaExoA (i.t.) + saline (i.v.). Frequency specific (2-20 kHz) auditory brainstem responses were measured to determine hearing thresholds before and 2, 5 and 10 days after instillation. RESULTS: PaExoA had penetrated from the middle ear into the cochlea and caused hearing loss. This impairment was blocked by intratympanic injection of edaravone when given simultaneously or 1 h after the first instillation of PaExoA, or by intravenous injection of edaravone when given simultaneously. There were significant differences in protective effect between the intratympanic and intravenous routes. PMID- 16308250 TI - Developing a predictive model for vertigo using demographic and laboratory data: An evidence-based medicine approach. AB - CONCLUSION: The approach described in this paper may be helpful in establishing an early-warning, evidence-based mechanism for diagnosing vertigo, which can be utilized in medical education to reduce medical uncertainty. OBJECTIVE: To use an evidence-based medicine approach to evaluate the probability of having vertigo using laboratory and demographic data. MATERIAL AND METHODS: The study was conducted on 22 working days during July 2002. Targeted cases who visited a general hospital in southern Taiwan for routine physical examinations were asked to participate in the study and agreed to take additional tests during their visits. A total of 200 subjects were systematically and randomly selected from this data pool. We ran binary logistic regression on all these cases. RESULTS: The logistic regression model explained 71.3% of the variance in having vertigo or not. The equation for having vertigo was as follows: -21.855 + (1.132 x male gender) + (0.071 x age) + (-0.023 x systolic blood pressure) + (0.057 x diastolic blood pressure) + (0.048 x fasting glucose) + (0.051 x cholesterol) + (-0.005 x triglycerides) + (-0.361 x presence of cardiovascular diseases). PMID- 16308251 TI - Differentiating the cause of acute sensorineural hearing loss between Meniere's disease and sudden deafness. AB - CONCLUSIONS: Most patients with Meniere's disease (MD) reveal abnormal vestibular evoked myogenic potentials (VEMPs) and the recruitment phenomenon, whereas most sudden deafness patients display normal VEMPs without the recruitment phenomenon. We therefore recommend using both the recruitment phenomenon and VEMP testing as a diagnostic algorithm to differentiate between MD and sudden deafness as the cause of acute hearing loss. OBJECTIVE: To recommend a diagnostic algorithm to differentiate between MD and sudden deafness as the cause of acute hearing loss. MATERIAL AND METHODS: Between January 2002 and December 2003, 14 consecutive patients with "probable" MD who also had acute sensorineural hearing loss were enrolled in the study, together with another 14 age- and sex-matched patients with idiopathic sudden deafness. Each patient underwent a battery of audiovestibular function tests, including pure-tone audiometry, a stapedial reflex test, distortion-product otoacoustic emissions (DPOAEs), electronystagmography and a VEMP test. All patients were followed for at least 12 months after presentation. RESULTS: Significant differences were found between MD and sudden deafness in terms of the recruitment phenomenon (86% and 21% of cases, respectively) and abnormal VEMP responses (71% and 21% of cases, respectively). However, the diseases did not differ significantly in terms of abnormal DPOAEs or caloric test results. PMID- 16308252 TI - Auditory pathway in rheumatoid arthritis. A comparative study and surgical perspectives. AB - CONCLUSION: Rheumatoid arthritis (RA) patients present with both conductive and sensorineural deafness. OBJECTIVE: To evaluate the prevalence and features of hearing impairment in patients with RA. MATERIAL AND METHODS: A total of 28 RA patients underwent a rheumatological evaluation, including determination of rheumatoid factor, protein 2-glycoprotein I level and the Lee index. An audiological assessment consisting of pure-tone audiometry (PTA) and determination of auditory brainstem responses (ABRs) and transient evoked otoacoustic emissions (TEOAEs) was performed. The results were compared with those of 28 age- and sex-matched healthy subjects. Four selected RA patients underwent stapedectomy; PTA and TEOAEs were evaluated 6 months postoperatively. RESULTS: Increased air conduction thresholds at 250, 500 and 1000 Hz were found in RA subjects in comparison to controls (p<0.001). RA patients showed higher air bone gaps in PTA (p<0.05) and an increased Wave I latency in ABRs (p=0.03). Decreased reproducibility (p<0.001) and amplitude (p<0.001) of TEOAEs were found in RA subjects in comparison to controls. A significant correlation between disease duration and echo amplitude was noticed (r=0.389). After stapedectomy, a reduction in the air-bone conduction gap (11 vs 2 dB HL) was noticed; no significant difference in TEOAEs was found. PMID- 16308253 TI - CT assessment of the effect of fluticasone propionate aqueous nasal spray treatment on lower turbinate hypertrophy due to vasomotor rhinitis. AB - CONCLUSION: Fluticasone propionate (FP) aqueous nasal spray was objectively found to be effective and safe for the treatment of lower turbinate enlargement in patients with vasomotor rhinitis. OBJECTIVE: To assess the efficacy of FP aqueous nasal spray treatment in lower turbinate hypertrophy due to vasomotor rhinitis using CT. MATERIAL AND METHODS: Of 35 patients with hypertrophic lower turbinates due to vasomotor rhinitis, 20 were treated twice daily with FP aqueous nasal spray (200 microg/day) for 3 months continuously and 15 were treated with placebo vehicle as a control group. The local effect of the nasal spray was studied using CT and visual analog scales. RESULTS: Treatment with FP provided significantly greater relief from the symptom of nasal obstruction compared with placebo over the entire 3-month treatment period (p < 0.001). When the change from baseline was compared between the two groups, FP produced statistically significant reductions in the mucosal area of the lower turbinates and in the thickness of the nasal mucosa after 3 months (p < 0.05). PMID- 16308254 TI - Expression of ecalectin, a novel eosinophil chemoattractant, in nasal polyps. AB - CONCLUSION: Ecalectin, which is produced in the mucosa of nasal polyps, seems to play an important role in the accumulation and activation of eosinophils in nasal polyps, regardless of the presence or absence of atopic predisposition. OBJECTIVE: Ecalectin is a recently discovered eosinophil chemoattractant which elongs to the galectin family. We investigated the expression of ecalectin in nasal polyp tissues associated with various nasal and paranasal diseases in order to clarify the pathogenesis of eosinophilia in nasal polyposis. MATERIAL AND METHODS: Nasal polyps were taken from 56 patients diagnosed as having chronic sinusitis with nasal polyposis. The surgically resected polyps and nasal turbinates were immunohistochemically stained using antibodies against EG2, human mast cell tryptase, CD3 and ecalectin. RESULTS: The number of EG2- and ecalectin positive cells was significantly higher in nasal polyps than control turbinates. Ecalectin-positive cells were observed in the subepithelial layer, where many EG2 positive cells were present. The number of ecalectin-positive cells correlated significantly with the number of EG2-positive cells in nasal polyps. Many ecalectin mRNA-positive cells were also observed in nasal polyps with an accumulation of EG2-positive cells. PMID- 16308255 TI - Adjuvant local anaesthetics in the epipharyngeal space in day-case adenoidectomy: a prospective, randomized, double-blind, placebo-controlled trial. AB - CONCLUSION: Mepivacaine applied in the epipharyngeal space as an adjuvant local anaesthetic does not reduce postoperative pain after adenoidectomy in children given high-dose paracetamol preoperatively. OBJECTIVE: To evaluate the pain reducing effect of an adjuvant local anaesthetic (mepivacaine) applied in the epipharyngeal space after outpatient adenoidectomy. MATERIAL AND METHODS: This was a prospective, randomized, double-blind, placebo-controlled trial. Postoperative pain was assessed in 98 children aged 3-10 years using a visual analogue scale (VAS) and the Wong-Baker FACES pain rating scale. The time between the end of surgery and discharge from hospital was recorded, as well as the need for extra pain medication in addition to the high-dose paracetamol (acetaminophen) that had been given preoperatively. Serum levels of mepivacaine were analysed in 10 consecutive patients. RESULTS: No difference was seen between the mepivacaine and control groups regarding postoperative pain as estimated using either the VAS or the Wong-Baker FACES pain rating scale. There were also no differences between the groups concerning the time to discharge from hospital and the need for extra pain medication before discharge. No mepivacaine could be detected in the patients' sera. PMID- 16308256 TI - Characteristic features of B cells in murine cervical lymph nodes. AB - CONCLUSION: B cells in cervical lymph nodes correspond to typical conventional B cells (B-2). OBJECTIVE: The special status of cervical lymph nodes in relation to the oropharynx, and the need to maintain the integrity of the oropharnygeal mucosal barrier, suggest the possibility that cervical lymph node B cells located in the oropharynx may behave differently from B cells located elsewhere. In this study we examined the symmetry or lack thereof between cervical lymph node B cells and other B-cell subsets. MATERIAL AND METHODS: We isolated B cells from murine cervical lymph node tissue and evaluated them in vitro according to several criteria. RESULTS: We found that cervical lymph node B cells expressed typical B-cell phenotypic markers and proliferated normally in response to mitogenic stimulation. They did not spontaneously secrete immunoglobulin and, in keeping with this, did not express elevated levels of either CD138 (Syndecan-1), a marker for plasma cells, or BLIMP-1, a putative master regulator of B-cell differentiation. PMID- 16308257 TI - Voice analysis of patients with diverse types of Reinke's edema and clinical use of electroglottographic measurements. AB - CONCLUSIONS: Reinke's edema shows a variety of clinical patterns and differences in voice analysis depending on its type. Electroglottographic measurements help to objectively assess the severity of edema and voice quality before and after surgery. OBJECTIVES: Reinke's edema manifests various clinical findings and different voice characteristics depending on the degree of swelling and the severity of voice impairment. The aim of this study was to compare the aerodynamic, acoustic and electroglottographic parameters for the different types of Reinke's edema according to its morphological, perceptual and histopathological classifications, and to assess voice quality in patients with Reinke's edema before and after laryngomicrosurgery. We also aimed to determine which parameter was most associated with the improvement in postoperative voice quality and to investigate the clinical usefulness of electroglottographic analysis in Reinke's edema. MATERIAL AND METHODS: The clinical and voice records of 61 patients with Reinke's edema were reviewed. All the patients were classified according to stroboscopic findings (Yonekawa's classification), perceptual grade and histopathological findings. Voice analysis of the perceptual, acoustic, aerodynamic and electroglottographic measurements was carried out using various classifications, and the voices of 23 patients who underwent laryngomicrosurgery were evaluated 2 months after surgery. Furthermore, the voices of 30 normal speakers (controls) were recorded and analyzed. RESULTS: The fundamental frequency (FxM) of Yonekawa Type III was significantly lower than those of the other types of Reinke's edema, and the SD of the FxM, the percentage irregularity of the FxM (CFx) and the percentage irregularity of the amplitude were larger than those of the other types of Reinke's edema. The closed quotient was significantly higher in Yonekawa Type III. In addition, the mean flow rate (MFR), maximum phonation time and harmonics:noise ratio (HNR) differed significantly among the different types of Reinke's edema. The postoperative results showed an increase in the FxM and an improvement in the MFR, subglottic pressure, shimmer and HNR. Correlation analysis showed that jitter, the HNR, the mean closed quotient and the irregularity of the frequency were the parameters that had the best correlation with improvement in postoperative voice quality. PMID- 16308258 TI - Role of lysosomal cathepsins in naphthazarin- and Fas-induced apoptosis in oral squamous cell carcinoma cells. AB - CONCLUSION: Intracellular cysteine cathepsins are pro-apoptotic factors involved in activation of caspases in two oral squamous cell carcinoma (SCC) cell lines. OBJECTIVE: To study the possible involvement of lysosomal cathepsins in oral SCC cell apoptosis. MATERIAL AND METHODS: Apoptosis was induced in the two human oral SCC cell lines UT-SCC-20A and UT-SCC-24A using naphthazarin or anti-Fas antibodies, and was studied by analysis of caspase activity and nuclear morphology. Involvement of lysosomal cathepsins was investigated using the cysteine cathepsin inhibitor z-FA-FMK and the cathepsin D inhibitor pepstatin A. The amounts of cellular and soluble Fas death receptor were determined by ELISA. RESULTS: Release of cathepsins from the lysosomes to the cytosol was observed early in apoptosis. Cysteine cathepsins were found to be involved in activation of caspases in response to treatment with naphthazarin or anti-Fas antibodies, but inhibition of cysteine cathepsin activity was not sufficient to prevent cell death. Moreover, inhibition of cysteine cathepsin activity resulted in increased expression of the Fas death receptor, suggesting involvement of extracellular cysteine cathepsins in death receptor shedding. PMID- 16308259 TI - Intraoperative computed tomography in otorhinolaryngology. AB - CONCLUSIONS: Intraoperative CT surgery provides the surgeon with additional information about the altered surgical site in difficult anatomical situations. The skull base and lamina papyracea may be revealed by means of intraoperative CT, which may be beneficial in endonasal sinus surgery involving difficult surgical sites, although individual ethmoid cells cannot be assessed owing to blood artefacts. This provides the surgeon with valuable information that may facilitate the procedure considerably. In soft-tissue surgery it is advisable to apply a contrast agent in order to achieve good soft-tissue contrast, thus allowing the tumour to be adequately distinguished from benign tissue. The intraoperative application of CT is a fairly time-consuming procedure, partly owing to the preparation time (set-up of the appliance; 10-min warming-up phase) and partly due to the length of time required to calculate each image (15 s). OBJECTIVE: CT is a well-established imaging method for the assessment of osseous and soft-tissue structures in the head and neck region. Saving information and transferring it to the intraoperative site may, however, be problematic. Computer assisted navigation systems are now able to assist difficult surgical procedures in the field of otolaryngology. To investigate the indications for intraoperative CT, we used it in various surgical procedures in the head and neck region. MATERIAL AND METHODS: Intraoperative CT was applied using the Tomoscan M in 46 cases in order to demonstrate the surgical benefit of the following procedures: endonasal surgical procedures on the paranasal sinuses (maxillary and ethmoidal sinusitis, anterior fracture of the sphenoidal sinus); tumour removal by means of laser surgery (carcinomas of the hypopharynx and larynx); and cochlear implantation (to verify the electrode position). After positioning the patient on the CT table, the workstation was set up in the operating theatre. If necessary, the gantry could be moved over the patient's head without repositioning the patient. RESULTS: Intraoperative CT was used to assist in the exposure of the skull base and lamina papyracea in endonasal surgery of the paranasal sinuses. Individual ethmoidal sinuses could not be evaluated owing to blood artefacts. Intraoperative imaging proved particularly helpful in revision surgery for chronic sinusitis in cases with missing anatomical landmarks owing to previous surgeries, where there is an increased risk of inflicting damage to the skull base or orbita. The resection margins can be determined in craniofacial resections. In soft-tissue procedures, such as tumour removal by means of laser surgery, it proved possible to visualize the resection borders of malignant tumours. Assessment of the electrode position in cochlear implantation is particularly useful in revision cases and in cases of cochlear obliteration. PMID- 16308260 TI - Expression of hepatocyte growth factor and c-Met in hypopharyngeal squamous cell carcinoma. AB - CONCLUSION: Our results suggest that hepatocyte growth factor (HGF) may play an important role in the progression of hypopharyngeal cancer. OBJECTIVE: HGF, a potent stimulator of hepatocyte growth, stimulates the motility, invasiveness, proliferation and morphogenesis of epithelium and may be involved in physiologic and pathologic processes such as embryogenesis, wound healing, organ regeneration, inflammation and tumor invasion. We therefore examined the role of HGF and c-Met in the invasion and metastasis of hypopharyngeal squamous cell carcinoma (SCC). MATERIAL AND METHODS: We performed immunohistochemical staining of 40 specimens each of normal mucosa and SCC of the hypopharynx with HGF and c Met antibodies. For reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting, fresh normal and cancer tissue from the hypopharynx obtained from five patients were used. RESULTS: Positive rates of HGF and c-Met expression in hypopharyngeal SCC were 77.5% and 70%, respectively. HGF staining was significantly correlated with lymph node metastasis and pathologic stage (p < 0.05). c-Met staining was only significantly correlated with lymph node metastasis (p < 0.05). Increased expression of c-Met mRNA (RT-PCR) and protein (Western blotting) was detected in hypopharyngeal cancer tissue. PMID- 16308261 TI - Facial nerve paralysis caused by middle ear cholesteatoma and effects of surgical intervention. AB - CONCLUSIONS: The clinical and surgical findings of this study indicated advanced cholesteatoma in many patients with facial paralysis. The outcome of facial paralysis was good. Poor outcomes were observed in cases with petrosal cholesteatoma and in those who underwent surgery > or = 2 months after the onset of paralysis. OBJECTIVE: To investigate clinical features of cholesteatoma associated with facial paralysis. MATERIAL AND METHODS: Sixteen patients with facial paralysis due to middle ear cholesteatoma were reviewed. After removal of the cholesteatoma lesion, a limited area of the fallopian canal, that in which facial nerve edema or redness was evident, was opened. Incision of the epineural sheath for nerve decompression was not performed. RESULTS: Initial paralysis was incomplete in 11 patients (69%). The onset of paralysis was sudden in 12 patients (75%). Labyrinthine fistulae (n = 9; 56%) and bone destruction in the cranial fossa (n = 10; 63%) were frequently observed. Six patients (38%) were totally deaf due to labyrinthitis. The outcome of facial paralysis was good in 13 patients (81%). Patients who underwent surgery > or = 2 months after the onset of paralysis frequently had a poor outcome. Paralysis was not improved in two cases with petrosal cholesteatoma. PMID- 16308262 TI - Euosmia: a rare form of parosmia. AB - The nature of qualitative olfactory disorders such as phantosmia and parosmia is a matter of debate. Parosmia and phantosmia mainly occur in combination with post traumatic or post-infectious olfactory loss. Rare causes of these disorders such as brain tumors, side-effects of drugs, paraneoplastic syndromes, psychiatric disorders or intracerebral haemorrhage have been reported. Parosmias are distorted sensations of smell elicited by an odor, whereas phantosmias persist permanently or occur without the presence of an odor source. Phantosmias differ widely in terms of their nature. In contrast, parosmias always seem to be unpleasant. We report the case of a female with post-infectious hyposmia who reported a pleasant parosmia to selected odorants. We have called this rare clinical presentation euosmia. PMID- 16308263 TI - A 15-year-old girl with a spindle cell carcinoma of the parotid gland. A diagnostic challenge solved by immunohistochemistry and cytogenetic analysis. AB - Spindle cell carcinoma is a rare neoplasm of the upper respiratory tract which occurs in adults, most commonly in the larynx. In the literature only one case of spindle cell carcinoma, located in the maxilla, has been reported in a child. We report the first presentation of a spindle cell carcinoma in a child, which was located in the parotid gland, together with the clinical course. The diagnostic challenge associated with this unusual disorder is elucidated, as well as the role of immunohistochemical and cytogenetic examination to define the nature of these lesions. PMID- 16308264 TI - Castleman's disease as an uncommon cause of a neck mass. AB - The detection of Castleman's disease in the neck is uncommon. However, it must be included in the differential diagnosis of a neck mass, together with other lymphoproliferative disorders or thyroid diseases. In 1999, three patients underwent resection of a neck mass of uncertain cytological diagnosis. Frozen section examinations and immunohistochemical studies using primary antibodies directed against B- and T-cell markers were performed. Very similar features were observed in all cases and the diagnosis of Castleman's disease was based on Frizzera's histological morphological criteria; all cases were of the hyaline vascular type. After surgical resection, the patients remained symptom-free after 51, 50 and 46 months, respectively, without clinical or radiographic signs of local or diffuse recurrence. Surgical removal of a mass of unknown origin located in the central part of the neck is mandatory because it allows a correct diagnosis to be made and is curative when the disease is limited to one site. PMID- 16308265 TI - Characterization of the biochemical effects of 1-nitronaphthalene in rats using global metabolic profiling by NMR spectroscopy and pattern recognition. AB - Metabolic fingerprints, in the form of patterns of high-concentration endogenous metabolites, of 1-nitronaphthalene (NN)-induced lung toxicity have been elucidated in bronchoalveolar lavage fluid (BALF), urine, blood plasma, and intact lung and liver tissue using NMR spectroscopy-based metabolic profiling. A single dose of NN (75 mg kg(-1)) was administered orally to Sprague-Dawley rats. BALF and lung tissue were obtained 24 h after dosing from these animals and matched control rats post-mortem. High-resolution (1)H-NMR spectroscopy of BALF samples indicated that NN caused increases in concentrations of choline, amino acids (leucine, isoleucine and alanine) and lactate together with decreased concentrations of succinate, citrate, creatine, creatinine and glucose. In addition, the intact lung weights were higher in the NN-treated group (p<0.01), consistent with pulmonary oedema. The NMR-detected perturbations indicated that NN induces a perturbation in energy metabolism in both lung and liver tissue, as well as surfactant production and osmolyte levels in the lungs. As well as reporting the first NMR spectroscopic combined examination of BALF and intact lung, this study indicates that such holistic approaches to investigating mechanisms of lung toxicity may be of value in evaluating disease progression or the effects of therapeutic intervention in pulmonary conditions such as surfactant disorders or asthma. PMID- 16308266 TI - Mytilus trossulus hsp70 as a biomarker for arsenic exposure in the marine environment: laboratory and real-world results. AB - The highly conserved heat shock protein 70 (hsp70) is induced by heat and chemical toxins, particularly heavy metals such as arsenic (As). The use of Mytilus trossulus (bay mussel) hsp70 as a 'screening' biomarker for marine heavy metals contamination was assessed. Some studies have found high hsp70 sensitivity to heavy metals, while others have found the opposite. Few studies have realistically used low heavy metals exposures, and fewer have used real-world contamination exposures. Clean sub-tidal mussels from the Puget Sound, Washington State (WA), USA, were acclimatized for 2 weeks and exposed for 24 h to As-spiked seawater (n=9) or to contaminated seawater from an arsenical pesticide plant in Tacoma, WA (n=10) followed by a Western blot for hsp70. Hsp70 inductions were insignificant at 10 microg l(-1) As(III), but were strong at 100 microg l(-1) (p<0.05) and 1000 microg l(-1) (p<0.01), with the induction threshold estimated at 30-50 microg l(-1) As(III). Hsp70 induction roughly correlated with arsenical toxicity, with As(III) > As(V) > (CH(3))(2)As(V). Altogether, the inter individual variability of hsp70 levels tends to mask inductions at low As concentrations, making it a crude toxicity biomarker. In addressing this problem, the following options could prove promising: (1) pre- or post-stressing specimens for greater hsp70 sensitivity, (2) use of internal protein controls such as actin, (3) use of hsp70-reporter gene constructs, and (4) detection with hsp60, heme oxygenase-1, metallothionein, CYP450, MXR or GPx. PMID- 16308267 TI - Quantification of vitellogenin mRNA induction in mosquitofish (Gambusia affinis) by reverse transcription real-time polymerase chain reaction (RT-PCR). AB - A method to quantify induction of vitellogenin (Vtg) mRNA in adult male mosquitofish was developed. Male mosquitofish were exposed to 0, 1, 20 and 250 ng l(-1) 17beta-oestradiol (E(2)) for 4 and 8 days in static exposures, and liver Vtg mRNA and 18S rRNA expression were quantified in duplex RT-PCR. Liver 18S rRNA expression was very consistent among individuals, and there was a highly significant increase in Vtg mRNA expression after exposure of mosquitofish for just 4 days at 250 ng l(-1) E(2). Lower doses did not induce Vtg mRNA expression even at 4 or 8 days. This method could be used as a rapid test to detect exposure of mosquitofish to oestrogenic chemicals. Further work is needed to determine if increased Vtg mRNA levels in male mosquitofish induce Vtg synthesis, and to determine the usefulness of the method in field sampling. PMID- 16308268 TI - Beta-naphthoflavone inhibits the induction of hepatic oestrogen-dependent proteins by 17alpha-ethynylestradiol in mosquitofish (Gambusia holbrooki). AB - The interactive effects of an aryl hydrocarbon receptor (AhR) agonist and of a xenoestrogen on biomarker responses were studied in the liver of male mosquitofish (Gambusia holbrooki). Hepatic 7-ethoxyresorufin O-deethylase (EROD) enzymatic activity was measured as a biomarker of exposure to the model AhR agonist beta-naphthoflavone (bNF). Hepatic proteins indicating the exposure of males to the synthetic oestrogen 17alpha-ethynylestradiol (EE2) were monitored by Western blot analysis using immunoserum prepared for this study. After a semi static exposure only to waterborne EE2, Western blot analysis of liver homogenate revealed the induction of two protein bands (a double band at 205 kDa and a single band at 125 kDa). The interaction between bNF and EE2 was investigated by analysing, on the one hand, EROD activity and, on the other hand, immunoreactivity corresponding to the two oestrogen-dependent protein bands in the liver of fish exposed to different concentrations of bNF for 2 days, then to the same concentrations of bNF plus 0.1 microg l(-1) EE2 for 5 days. EE2 changed neither the basal activity of EROD nor its rate of induction with 1.0 and 4.0 microg l(-1) bNF. On the other hand, the induction of oestrogen-dependent proteins with 0.1 microg l(-1) EE2 was inhibited by exposure to 4.0 microg l(-1) bNF. These results together with literature data suggest that field monitoring of xenoestrogen contamination through the analysis of oestrogen-dependent protein in male fish as a biomarker should take into account the possible negative interference of AhR agonists. PMID- 16308269 TI - Increased serum bile acids as a possible biomarker of hepatotoxicity in Brazilian workers exposed to solvents in car repainting shops. AB - The objective was to evaluate total serum bile acids (SBA) as a biological marker of hepatotoxicity in car painters exposed to organic solvents and to compare their performance with classic biochemical parameters of liver function. SBA were analysed in a selected group of workers (n=57) occupationally exposed to a mixture of organic solvents and in a control group (n=51). In addition, aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TB), gamma-glutamyltransferase (GGT) and alkaline phosphatase (ALP) were determined in the two groups. Urinary hippuric acid was measured in all samples. Statistical analysis of the data revealed a significant increase in the concentration of SBA, AST, ALP and TB in exposed workers compared with controls (Mann-Whitney, p/=5 ppm) group and 2.41 ppm (range 0.9-4.3 ppm) in the low-exposure (<5 ppm) group. There were 13 of 44 (29.6%) abnormal liver function tests (elevations of either glutamate oxaloacetate transaminase (GOT) or glutamate pyruvate transaminase (GPT)) among the high DMF exposure workers, two of 22 (9.1%) abnormal liver function tests among the low DMF exposure workers. Chronic liver disease as determined by ultrasonography was present in seven of 44 (15.9%) high DMF exposure workers, and 0 of 22 (0%) low DMF exposure workers. There were 11 of 34 (32.4%) abnormal liver function tests among the GSTT1 null genotype workers, and four of 32 (12.5%) abnormal liver function tests among the GSTT1-positive genotype workers. Compared with the low DMF exposure workers, the adjusted odds ratio and 95% confidence intervals for abnormal liver function tests was 6.78 (0.94-48.7) for the high DMF exposure workers. Compared with the GSTT1-positive genotype workers, the adjusted odds ratio and 95% confidence intervals for abnormal liver function tests was 4.41 (1.15-16.9) for the GSTT1 null genotype workers. Compared with the low DMF group with GSTT1-positive genotype workers, the odds ratio (adjusted for HBV status) of abnormal liver function test was 12.38, 95% CI=(1.04-146.9) for the high DMF group with GSTT1 null genotype workers. This study indicates that abnormal liver function and chronic liver disease are associated with DMF exposure, and there are more than multiplicative interaction effects on abnormal liver function tests between the DMF exposure and the GSTT1 genotype. PMID- 16308271 TI - Changes in serological biomarkers of liver function and connective tissue turnover in chronic hepatitis B during lamivudine therapy. AB - Assessment of hepatic damage associated with chronic hepatitis B (CHB) currently relies on measurement of serum transaminases and assessment of hepatic histology. It was determined serum hepatic function tests and the liver fibrosis biomarkers type IV collagen (CIV), amino-terminal propeptide of type I procollagen (PINP), amino-terminal propeptide of type III procollagen (PIIINP) and carboxy-terminal telopeptide of type I collagen (ICTP) were of value in monitoring the effect of lamivudine therapy for CHB. Thirty-nine patients received orally 100 mg lamivudine daily for 48 weeks. Blood samples were obtained at baseline, 24 and 48 weeks. At the end of the treatment period, the patients were then divided into four groups according to the pattern of HBs and HBe antigens. At baseline, alanine aminotransferase, aspartate aminotransferase, PIIINP and the PINP/ICTP ratio and at 24 weeks alanine aminotransferase, aspartate aminotransferase and the PINP/ICTP ratio had lower values in the complete response compared with complete failure groups. Using receiver-operated curve analysis, only the PINP/ICTP ratio at baseline (area under the curve 0.806) and ALT and the PINP/ICTP ratio at 24 weeks (areas under the curve 0.803 and 0.776, respectively) had significant diagnostic ability in detecting responders. In conclusion, the PINP/ITCP ratio is sensitive and specific in detecting responders to treatment. PMID- 16308272 TI - Membrane-bound ARF1 peptide: interpretation of neutron diffraction data by molecular dynamics simulation methods. AB - Adenosine diphosphate ribosylation factor-1 (ARF1) is activated by cell membrane binding of a self-folding N-terminal domain. We have previously presented four possible conformations of the membrane bound, human ARF1 N-terminal peptide in planar lipid bilayers of DOPC and DOPG (7:3 molar ratio), determined from lamellar neutron diffraction and circular dichroism data. In this paper we analyse the four possible conformations by molecular dynamics simulations. The aim of these simulations was to use MD to distinguish which of the four possible membrane bound structures was the most likely. The most likely conformation was determined according to the following criteria: (a) location of label positions on the peptide in relation to the bilayer, (b) lowest mean square displacement from the initial structure, (c) lowest system energy, (d) most peptide-lipid headgroup hydrogen bonding, (e) analysis of phi/psi angles of the peptide. These findings demonstrate the application of molecular dynamics simulations to explore neutron diffraction data. PMID- 16308273 TI - N type rapid inactivation in human Kv1.4 channels: functional role of a putative C-terminal helix. AB - Voltage gated potassium channels are tetrameric membrane proteins, which have a central role in cellular excitability. Human Kv1.4 channels open on membrane depolarization and inactivate rapidly by a 'ball and chain' mechanism whose molecular determinants have been mapped to the cytoplasmic N terminus of the channel. Here we show that the other terminal end of the channel also plays a role in channel inactivation. Swapping the C-terminal residues of hKv1.4 with those from two non-inactivating channels (hKv1.1 and hKv1.2) affects the rates of inactivation, as well as the recovery of the channel from the inactivated state. Secondary structure predictions of the hKv1.4 sequence reveal a helical structure at its distal C-terminal. Complete removal or partial disruption of this helical region results in channels with remarkably slowed inactivation kinetics. The ionic selectivity and voltage-dependence of channel opening were similar to hKv1.4, indicative of an unperturbed channel pore. These results demonstrate that fast inactivation is modulated by structural elements in the C-terminus, suggesting that the process involves the concerted action of the N- and C termini. PMID- 16308274 TI - Intracellular interaction between syntaxin and Munc 18-1 revealed by fluorescence resonance energy transfer. AB - Neurosecretion is catalyzed by assembly of a soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE)-complex composed of SNAP-25, synaptobrevin and syntaxin. Munc 18-1 is known to bind to syntaxin in vitro. This interaction prevents assembly of the SNARE-complex, but might also affect intracellular targeting of the proteins. We have fused syntaxin and Munc 18 to the yellow- (YFP) or cyan-fluorescence-protein (CFP) and expressed the constructs in CHO- and MDCK-cells. We have studied their localization with confocal microscopy and a possible protein-protein interaction with fluorescence-resonance energy transfer (FRET). YFP-syntaxin localizes to intracellular membranes. CFP Munc 18 is present in the cytoplasm as expected for a protein lacking membrane targeting domains. However, Munc 18 is redirected to internal membranes when syntaxin is coexpressed, but only limited transport of the proteins to the plasma membrane was observed. An interaction between Munc 18 and syntaxin could be demonstrated by FRET using two methods, sensitized acceptor fluorescence and acceptor photobleaching. A mutation in syntaxin (L165A, E166A), which is known to inhibit binding to Munc 18 in vitro, prevents colocalization of the proteins and also the FRET signal. Thus, a protein-protein interaction between Munc 18 and syntaxin occurs on intracellular membranes, which is required but not sufficient for quantitative transport of both proteins to the plasma membrane. PMID- 16308275 TI - Band 3 tyr-phosphorylation in normal and glucose-6-phospate dehydrogenase deficient human erythrocytes. AB - Haemolysis is usually episodic in glucose-6-phosphate dehydrogenase (G6PD) deficiency, often triggered by a period of oxidative stress. In the present work, we investigate a possible biochemical mechanism underlying the enhanced susceptibility of G6PD deficient red blood cells (RBC) to oxidative stress. We analysed eight male subjects with Mediterranean glucose-6P-dehydrogenase deficiency (G6PDd), class II, for their ability in phosphorylating erythrocyte membrane band 3 following oxidative and osmotic stress. Our findings show that this sensitivity is connected to an early membrane band 3 Tyr-phosphorylation in the presence of diamide. However, since both Syk, and Lyn kinases, and SHP-2 phosphatase, mostly implicated in the band 3 P-Tyr level regulation, are alike in content and activity in normal and patient erythrocytes, an alteration in the membrane organization is likely the cause of the anomalous response to the oxidant. We report, in fact, that hypertonic-induced morphological change in G6PDd erythrocyte induces a higher membrane band 3 Tyr-phosphorylation, suggesting a pre-existing membrane alteration, likely due to the chronic lowering of the redox systems in patients. We also report that 1-chloro-2,4-dinitrobenzene pre-treatment of normal red cells can alter the normal protein-protein and protein-membrane interaction under hypertonic rather than oxidative stress, thus partially resembling the response in patients, and that RBC may utilize a wider range of redox defence, under oxidative conditions, including, but not exclusively, NADPH and glutathione. On the whole, these results would encourage a different approach to the evaluation of the effects of pharmacological administration to patients, giving more attention to the possible drug-induced membrane alteration evidenced by the abnormal band 3 Tyr-phosphorylation. PMID- 16308276 TI - Association of small ankyrin 1 with the sarcoplasmic reticulum. AB - Small ankyrin 1, or sAnk1, is a small, alternatively spliced product of the erythroid ankyrin gene, ANK1, that is expressed in striated muscle and concentrated in the network sarcoplasmic reticulum (SR) surrounding the Z disks and M lines. We have characterized sAnk1 in muscle homogenates and SR vesicles, and have identified the region that targets it to the network SR. Selective extractions and partitioning into Triton X-114 show that sAnk1 behaves like the SR Ca-ATPase and so is an integral protein of the SR membrane. Mild proteolytic treatment of isolated SR vesicles indicates that sAnk1 is oriented with its hydrophilic, C-terminal sequence exposed to the solution, which is equivalent to the cytoplasmic face of the SR membrane in situ. SDS-PAGE in non-reducing gels suggests that sAnk1 is present as dimers and larger oligomers in the native SR. These results suggest that sAnk1 is oligomeric and oriented with its C-terminus exposed to the cytoplasm, where it may interact with proteins of the contractile apparatus. The N-terminal 29 amino acid hydrophobic sequence of sAnk1, which is predicted to span the SR membrane, is sufficient to target proteins to and anchor them in internal membranes of HEK 293 cells. It also targets reporter proteins to the network SR of skeletal myofibers and is thus the first example of a sequence that targets proteins to a particular compartment of the SR. PMID- 16308277 TI - Binding of antibiotic amphotericin B to lipid membranes: monomolecular layer technique and linear dichroism-FTIR studies. AB - Amphotericin B (AmB) is one of the main antibiotics applied in treatment of deep seated mycotic infections. Tensiometric technique has been applied to monitor binding of AmB, from the water subphase, to the lipid monomolecular layers, formed with dipalmitoylphosphatidylcholine at the air-water interface. Time dependencies of surface pressure in the monolayers demonstrate strong enhancement of AmB binding to monolayers brought about by sterols present in the membranes. The monolayers have been deposited to a solid support and examined by means of FTIR spectroscopy. FTIR measurements show that majority of the AmB molecules which bind to the membranes are localized in the polar headgroup region. The results of the linear dichroism-FTIR measurements are consistent with the microscopic picture according to which the molecules of the membrane-bound AmB are distributed among two orientational fractions: one horizontal and one vertical with respect to the plane of the membrane (59% versus 41% respectively, in the case of the membrane formed with the pure lipid without sterols). The presence of cholesterol in the membranes (50 mol% with respect to lipid) slightly affects such a distribution (53% horizontal versus 47% vertical) but the presence of ergosterol has a pronounced effect in the increase in population of the fraction of horizontally bound AmB (85% horizontal vs. 15% vertical). The results of the measurements indicate that mode of action of the AmB consists in disruption of the polar headgroup region of biomembranes, brought about by the AmB molecules bound horizontally with respect to the plane of the membrane. PMID- 16308278 TI - Substrate-induced conformational changes of the mitochondrial oxoglutarate carrier: a spectroscopic and molecular modelling study. AB - The structural and dynamic properties of the oxoglutarate carrier were investigated by introducing a single tryptophan in the Trp-devoid carrier in position 184, 190 or 199 and by monitoring the fluorescence spectra in the presence and absence of the substrate oxoglutarate. In the absence of substrate, the emission maxima of Arg190Trp, Cys184Trp and Leu199Trp are centered at 342, 345 and 348 nm, respectively, indicating that these residues have an increasing degree of solvent exposure. The emission intensity of the Arg190Trp and Cys184Trp mutants is higher than that of Leu199Trp. Addition of substrate increases the emission intensity of Leu199Trp, but not that of Cys184Trp and Arg190Trp. A 3D model of the oxoglutarate carrier was built using the structure of the ADP/ATP carrier as a template and was validated with the experimental results available in the literature. The model identifies Lys122 as the most likely candidate for the quenching of Trp199. Consistently, the double mutant Lys122Ala-Leu199Trp exhibits a higher emission intensity than Leu199Trp and does not display further fluorescence enhancement in response to substrate addition. Substitution of Lys122 with Cys and evaluation of its reactivity with a sulphydryl reagent in the presence and absence of substrate confirms that residue 122 is masked by the substrate, likely through a substrate-induced conformational change. PMID- 16308280 TI - Catalytic activities of human cytochrome P450 2C9*1, 2C9*3 and 2C9*13. AB - Cytochrome P450 2C9 (CYP2C9) is a geneticly polymorphic enzyme responsible for the metabolism of some clinically important drugs. CYP2C9*13 is an allele identified in a Chinese poor metabolizer of lornoxicam which has a Leu90Pro amino acid substitution. This paper reports on a study aimed at comparing the catalytic properties of CYP2C9*13 with those of the wild-type CYP2C9*1 and mutant CYP2C9*3 (Ile359Leu) in the COS-7 expression system using various substrates. CYP2C9*3 and *13 produced far lower luminescence than CYP2C9*1 in luciferin H metabolism. CYP2C9*13 exhibited an 11-fold increase in Km but no change in Vmax with tolbutamide as the substrate, a five-fold increase in Km and an 88.8% reduction in Vmax with diclofenac. These data indicate that CYP2C9*13 exhibits reduced metabolic activity toward all studied CYP2C9 substrates. The magnitude of the CYP2C9*13-associated decrease in intrinsic clearance (Vmax/Km) is greater than that associated with CYP2C9*3. PMID- 16308281 TI - In vivo induction of human cytochrome P450 3A4 by rifabutin in chimeric mice with humanized liver. AB - The induction of human cytochrome P450 enzymes (CYPs) often poses a serious problem in clinical practice. The induction of CYP3A leads to a decrease in the pharmacological potency of drugs, since many drugs are substrates of CYP3A. The present study examined the in vivo induction potency of human CYP3A in chimeric mice with humanized liver, recently established in Japan, by a specific inducer of human CYP3A enzyme activity in this experimental condition, rifabutin, which is an analogue of rifampicin. The chimeric mice were treated intraperitoneally daily for 4 days with rifabutin (50 mg kg(-1) day(-1)). The mRNA, protein and enzyme activity in liver of the chimeric mice were measured by reverse transcriptase polymerase chain reaction, Western blot analysis and high performance liquid chromatography, respectively. In the chimeric mice, the human CYP3A4 mRNA expression, CYP3A4 protein content, testosterone 6ss-hydroxylase activity and dexamethasone 6-hydroxylase activity were increased 7.4-, 3.0-, 2.4- and 1.9-fold, respectively, by treatment with rifabutin. The mRNA expression of other human CYPs, transporters and nuclear receptors was not significantly changed by rifabutin. On the other hand, rifabutin was demonstrated not to increase the murine Cyp3a enzyme activities in the control mice. It was demonstrated that human CYP3A4 expressed in the chimeric mice with humanized liver was induced by rifabutin, suggesting that human CYP3A4 in the chimeric mice had induction potency. This chimeric mouse model may be a useful animal model to estimate and predict the in vivo induction of CYPs in human. PMID- 16308282 TI - Evaluation of mRNA expression of human drug-metabolizing enzymes and transporters in chimeric mouse with humanized liver. AB - The hepatic mRNA expression of human drug-metabolizing enzymes and transporters in chimeric mise with almost-completely humanized liver (replacement index: 71 89%) was investigated. The mRNAs of 58 human phase I enzymes, 26 human phase II enzymes, 23 human transporters, and five mouse Cyps were measured in the chimeric mice with humanized liver generated using hepatocytes from a Japanese donor. The mRNA expression of 52 human phase I enzymes, which includes 20 human CYPs, 26 human phase II enzymes and 21 human transporters was ascertained in the chimeric mouse liver. Among them, the expression of the target mRNAs vital for liver function such as the metabolism and secretion of endogenous compounds appeared to be maintained. The central value for the expression ratio in all target genes in chimeric mouse liver to the donor liver was 0.46, which was lower than the substitution rate of chimeric mouse liver by donor liver. The ratio of mouse Cyp mRNA expression of chimeric mouse liver to that of control mouse liver was 0.19 or less, except for that of Cyp2b10. There were good correlations between the mRNA expression levels of human hepatic albumin gene, the values of the rate of replacement of mouse liver by human liver, and the human blood albumin concentration in the chimeric mice. The chimeric mice with humanized liver may be a useful tool for the evaluation of drug-drug interactions such as the inhibition and induction of drug-metabolizing enzymes and transporters. PMID- 16308283 TI - Pharmacokinetics of BAY 59-7939--an oral, direct Factor Xa inhibitor--in rats and dogs. AB - The pharmacokinetics of BAY 59-7939 - a novel, oral, direct Factor Xa inhibitor - were investigated in rats and dogs in support of preclinical safety studies and clinical development. BAY 59-7939 was rapidly absorbed after oral dosing, with an absolute bioavailability of 57-66% in rats, and 60-86% in dogs. Plasma pharmacokinetics of BAY 59-7939 were linear across the investigated dose range (1 10 mg kg(-1) in rats, 0.3-3 mg kg(-1) in dogs). Plasma clearance was low: 0.4 l kg(-1) h(-1) in rats and 0.3 l kg(-1) h(-1) in dogs; volume of distribution (V(ss)) was moderate: 0.3 l kg(-1) in rats, and 0.4 l kg(-1) in dogs. The elimination half-life after oral administration was short in both species (0.9 2.3 h). Whole-body autoradiography showed moderate tissue affinity. No retention or small volume enrichments of BAY 59-7939-related radioactivity were observed. The plasma-protein binding of BAY 59-7939 was high, species dependent and fully reversible. BAY 59-7939 was rapidly excreted in rats and dogs, and was not irreversibly retained. A dual mode of excretion (biliary/faecal and renal) was observed. In summary, BAY 59-7939 had a favourable, predictable pharmacokinetic profile, with high oral bioavailability and a dual route of excretion. PMID- 16308284 TI - Antigenicity of sulfanilamide and its metabolites using fluorescent-labelled compounds. AB - In order to clarify the onset mechanisms of drug-induced allergies, three fluorescent-labelled compounds were synthesized by subjecting sulfanilamide (SA), a base compound for sulfonamides, and its active metabolites, i.e. sulfanilamide hydroxylamine and sulfanilamide nitroso, to dansylation using dansylchloride. In other words, 5-dimethylamino-N-(4-aminobenzyl)-naphthalenesulfonamide (DNS-4ABA), 5-dimethylamino-N-(4-hydroxylaminobenzyl)-1-naphthalenesulfonamide (DNS-4HABA) and 5-dimethylamino-N-(4-nitrosobenzyl)-1-naphthalenesulfonamide (DNS-4NSBA) were synthesized as model haptens. When analysed by HPLC, a conjugate of DNS-4HABA and glutathione (GSH) with nucleophilic amino acids had two peaks (P-1 and P-2). FAB MS and 1H-NMR revealed that the DNS-4HABA-GSH conjugate consisted of sulphinamide and semimercaptal. The reactivity of GSH to DNS-4ABA, DNS-4HABA and DNS-4NSBA was quantified by HPLC using an oxidization system (horseradish peroxidase/H2O2). The results show that production of DNS-4NSBA-GSH-conjugate was four to eight times higher than that of DNS-4HABA-GSH conjugate, but that DNS-4ABA did not bind with GSH. Skin reactions were assessed using guinea pigs, and strong delayed erythema was seen with DNS-4NSBA, which bound most strongly with GSH, whereas weak delayed erythema was seen with DNS-4ABA, which did not bind with GSH. This suggests a correlation between GSH conjugate production and skin reactions. DNS-4HABA enzymatically bound with proteins in rat and guinea pig liver cytosol and microsomal fractions. The proteins that bound to DNS-4HABA were purified by HPLC and then subjected to N-terminal amino acid analysis. Ubiquitin (10 kDa) and fatty acid binding protein (30 kDa) were detected in the rat liver cytosol fraction; retinol-dehydrogenase (35 kDa) in the rat microsomal fraction; and glutathione-S-transferase B (mmu) (25 kDa) in the guinea pig liver cytosol fraction. When DNS-4HABA or DNS-4NSBA binds to proteins that play important roles in the body, unexpected adverse reactions may occur. Furthermore, by utilizing our technique using model compounds, it may be possible to identify the carrier proteins of various compounds, including pharmaceutical agents. PMID- 16308312 TI - Involvement of p38 MAPK and Nrf2 in phenolic acid-induced P-form phenol sulfotransferase expression in human hepatoma HepG2 cells. AB - Phenolic acids have significant biological and pharmacological properties and some have demonstrated remarkable ability to alter sulfate conjugation. However, the modulation mechanisms of phenolic acids on phenol sulfotransferase expression have not been described. In the present study, we investigated the effects of phenolic acids on the expression of the Phase II P-form of phenol sulfotransferase (PST-P) in human hepatoma HepG2 cells. RT-PCR and western blot data revealed that gallic acid induced increase in PST-P expression at the mRNA and protein levels, respectively. This induction was also marked by an increase in PST-P activity. Actinomycin D and cycloheximide inhibited gallic acid responsive PST-P mRNA expression, indicating that gallic acid is a requirement for transcription and de novo protein synthesis. Transient transfection of HepG2 cells with a reporter plasmid of the upstream region of the human PST gene caused a significant increase in reporter gene activity after gallic acid exposure. Moreover, gallic acid increased the nuclear levels of Nrf2, a transcription factor governing antioxidant response element (ARE). Electrophoretic mobility shift assay showed increased binding of nuclear proteins to ARE consensus sequence after treatment with gallic acid. While investigating the signaling pathways responsible for PST-P induction, we observed that gallic acid activated the p38 mitogen-activated protein kinase (MAPK) pathway. SB203580, a specific inhibitor of p38 MAPK, abolished gallic acid-induced PST-P protein expression. Similarly, gallic acid also caused an accumulation of Nrf2. Moreover, the protective effects of gallic acid on tert-butyl hydroperoxide-induced toxicity was partially blocked by p38 MAPK and PST-P inhibitors, further demonstrating that gallic acid attenuates oxidative stress through a pathway that involves p38 MAPK and PST-P. These results indicate that gallic acid is a potent inducer of PST-P and that PST-P induction is responsible for the gallic acid-mediated cytoprotection against oxidative damage. PMID- 16308313 TI - DNA repair polymorphisms and cancer risk in non-smokers in a cohort study. AB - Environmental carcinogens contained in air pollution, such as polycyclic aromatic hydrocarbons, aromatic amines or N-nitroso compounds, predominantly form DNA adducts but can also generate interstrand cross-links and reactive oxygen species. If unrepaired, such lesions increase the risk of somatic mutations and cancer. Our study investigated the relationships between 22 polymorphisms (and their haplotypes) in 16 DNA repair genes belonging to different repair pathways in 1094 controls and 567 cancer cases (bladder cancer, 131; lung cancer, 134; oral-pharyngeal cancer, 41; laryngeal cancer, 47; leukaemia, 179; death from emphysema and chronic obstructive pulmonary disease, 84). The design was a case control study nested within a prospective investigation. Among the many comparisons, few polymorphisms were associated with the diseases at the univariate analysis: XRCC1-399 Gln/Gln variant homozygotes [odds ratios (OR) = 2.20, 95% confidence intervals (CI) = 1.16-4.17] and XRCC3-241 Met/Met homozygotes (OR = 0.51, 95% CI = 0.27-0.96) and leukaemia. The recessive model in the stepwise multivariate analysis revealed a possible protective effect of XRCC1 399Gln/Gln in lung cancer (OR = 0.22, 95% CI = 0.05-0.98), and confirmed an opposite effect (OR = 2.47, 95% CI = 1.02-6.02) in the leukaemia group. Our results also suggest that the XPD/ERCC1-GAT haplotype may modulate leukaemia (OR = 1.28, 95% CI = 1.02-1.61), bladder cancer (OR = 1.38, 95% CI = 1.06-1.79) and possibly other cancer risks. Further investigations of the combined effects of polymorphisms within these DNA repair genes, smoking and other risk factors may help to clarify the influence of genetic variation in the carcinogenic process. PMID- 16308314 TI - Delphinidin, a dietary anthocyanidin, inhibits vascular endothelial growth factor receptor-2 phosphorylation. AB - Epidemiological studies have shown that a diet rich in fruits and vegetables has a beneficial preventive effect on cardiovascular diseases and cancer by mechanisms that have not yet been elucidated. In this work, we investigated the antiangiogenic activities of anthocyanidins, a class of polyphenols present at high levels in fruits. Among the tested anthocyanidins (cyanidin, delphinidin, malvidin, pelargonidin, peonidin and petunidin), delphinidin was the most potent angiogenic inhibitor. In vitro, low concentrations of delphinidin inhibited vascular endothelial growth factor (VEGF)-induced tyrosine phosphorylation of VEGF receptor (VEGFR)-2, leading to the inhibition of downstream signaling triggered by VEGFR-2. Inhibition of VEGFR-2 by delphinidin inhibited the VEGF induced activation of ERK-1/2 signaling and the chemotactic motility of human EC as well as their differentiation into capillary-like tubular structures in Matrigel and within fibrin gels. In vivo, delphinidin was able to suppress basic fibroblast growth factor-induced vessel formation in the mouse Matrigel plug assay. The identification of delphinidin as a naturally occurring inhibitor of VEGF receptors suggests that this molecule possesses important antiangiogenic properties that may be helpful for the prevention and treatment of cancer. PMID- 16308315 TI - Functional inactivation of p53 by human T-cell leukemia virus type 1 Tax protein: mechanisms and clinical implications. AB - Human T-cell leukemia virus type 1 (HTLV-I) has been implicated with the etiology of adult T-cell leukemia (ATL) and certain other clinical disorders. Although the leukemogenic mechanism of HTLV-1 is not fully understood yet, the viral Tax protein is widely regarded as a key factor in this mechanism. Tax can modulate the synthesis or function of many regulatory factors which control a wide range of normal and oncogenic cellular processes and therefore, it acts as a potent oncoprotein. In the last few years, special attention has been attracted to Tax interference with the transactivation function of p53, a tumor-suppressor protein that is involved in regulation of the cell-cycle and apoptosis and in maintaining the cellular genome integrity. p53 is mutated in approximately 60% of all human tumors. In contrast, mutant p53 is found in only small percentage of ATL patients. Nevertheless, p53 is inactive in the leukemic cells of most ATL patients and in most HTLV-1 transformed cells. By inactivating p53, Tax can immortalize the HTLV-1-infected cells and destabilize their genome. Consequently, such cells can progress toward the ultimate leukemic state by a stepwise accumulation of oncogenic mutations and other types of chromosomal aberrations. Furthermore, since p53 exists in most ATL patients in its wild-type form, its reactivation by therapeutic drugs might be an effective approach for ATL therapy. Several mechanisms have been proposed so far for Tax-induced p53 inactivation. Understanding the exact mechanism of this Tax effect is essential for designing effective means for this therapeutic approach. In this review article, we discuss the various mechanisms proposed for Tax interference with p53 functions and their clinical and therapeutic implications. PMID- 16308316 TI - Crystal structure of a bifunctional deaminase and reductase from Bacillus subtilis involved in riboflavin biosynthesis. AB - Bacterial RibG is an attractive candidate for development of antimicrobial drugs because of its involvement in the riboflavin biosynthesis. The crystal structure of Bacillus subtilis RibG at 2.41-A resolution displayed a tetrameric ring-like structure with an extensive interface of approximately 2400 A(2)/monomer. The N terminal deaminase domain belongs to the cytidine deaminase superfamily. A structure-based sequence alignment of a variety of nucleotide deaminases reveals not only the unique signatures in each family member for gene annotation but also putative substrate-interacting residues for RNA-editing deaminases. The strong structural conservation between the C-terminal reductase domain and the pharmaceutically important dihydrofolate reductase suggests that the two reductases involved in the riboflavin and folate biosyntheses evolved from a single ancestral gene. Together with the binding of the essential cofactors, zinc ion and NADPH, the structural comparison assists substrate modeling into the active-site cavities allowing identification of specific substrate recognition. Finally, the present structure reveals that the deaminase and the reductase are separate functional domains and that domain fusion is crucial for the enzyme activities through formation of a stable tetrameric structure. PMID- 16308317 TI - Structure of Chlorobium tepidum sepiapterin reductase complex reveals the novel substrate binding mode for stereospecific production of L-threo tetrahydrobiopterin. AB - Sepiapterin reductase (SR) is involved in the last step of tetrahydrobiopterin (BH(4)) biosynthesis by reducing the di-keto group of 6-pyruvoyl tetrahydropterin. Chlorobium tepidum SR (cSR) generates a distinct BH(4) product, L-threo-BH(4) (6R-(1'S,2'S)-5,6,7,8-BH(4)), whereas animal enzymes produce L erythro-BH(4) (6R-(1'R,2'S)-5,6,7,8-BH(4)) although it has high amino acid sequence similarities to the other animal enzymes. To elucidate the structural basis for the different reaction stereospecificities, we have determined the three-dimensional structures of cSR alone and complexed with NADP and sepiapterin at 2.1 and 1.7 A resolution, respectively. The overall folding of the cSR, the binding site for the cofactor NADP(H), and the positions of active site residues were quite similar to the mouse and the human SR. However, significant differences were found in the substrate binding region of the cSR. In comparison to the mouse SR complex, the sepiapterin in the cSR is rotated about 180 degrees around the active site and bound between two aromatic side chains of Trp-196 and Phe-99 so that its pterin ring is shifted to the opposite side, but its side chain position is not changed. The swiveled sepiapterin binding results in the conversion of the side chain configuration, exposing the opposite face for hydride transfer from NADPH. The different sepiapterin binding mode within the conserved catalytic architecture presents a novel strategy of switching the reaction stereospecificities in the same protein fold. PMID- 16308318 TI - Suppression of RhoA activity by focal adhesion kinase-induced activation of p190RhoGAP: role in regulation of endothelial permeability. AB - The interaction of endothelial cells with extracellular matrix proteins at focal adhesions sites contributes to the integrity of vascular endothelial barrier. Although focal adhesion kinase (FAK) activation is required for the recovery of the barrier function after increased endothelial junctional permeability, the basis for the recovery remains unclear. We tested the hypothesis that FAK activates p190RhoGAP and, thus, negatively regulates RhoA activity and promotes endothelial barrier restoration in response to the permeability-increasing mediator thrombin. We observed that thrombin caused a transient activation of RhoA but a more prolonged FAK activation temporally coupled to the recovery of barrier function. Thrombin also induced tyrosine phosphorylation of p190RhoGAP, which coincided with decrease in RhoA activity. We further showed that FAK was associated with p190RhoGAP, and importantly, recombinant FAK phosphorylated p190RhoGAP in vitro. Inhibition of FAK by adenoviral expression of FRNK (a dominant negative FAK construct) in monolayers prevented p190RhoGAP phosphorylation, increased RhoA activity, induced actin stress fiber formation, and produced an irreversible increase in endothelial permeability in response to thrombin. We also observed that p190RhoGAP was unable to attenuate RhoA activation in the absence of FAK activation induced by FRNK. The inhibition of RhoA by the C3 toxin (Clostridium botulinum toxin) restored endothelial barrier function in the FRNK-expressing cells. These findings in endothelial cells were recapitulated in the lung microcirculation in which FRNK expression in microvessel endothelia increased vascular permeability. Our studies demonstrate that FAK-induced down-modulation of RhoA activity via p190RhoGAP is a crucial step in signaling endothelial barrier restoration after increased endothelial permeability. PMID- 16308319 TI - Heterogeneous nuclear ribonucleoprotein H is required for optimal U11 small nuclear ribonucleoprotein binding to a retroviral RNA-processing control element: implications for U12-dependent RNA splicing. AB - An RNA-processing element from Rous sarcoma virus, the negative regulator of splicing (NRS), represses splicing to generate unspliced RNA that serves as mRNA and as genomic RNA for progeny virions and also promotes polyadenylation of the unspliced RNA. Integral to NRS function is the binding of U1 small nuclear ribonucleoprotein (snRNP), but its binding is controlled by U11 snRNP that binds to an overlapping site. U11 snRNP, the U1 counterpart for splicing of U12 dependent introns, binds the NRS remarkably well and requires G-rich elements just downstream of the consensus U11 binding site. We present evidence that heterogeneous nuclear ribonucleoprotein (hnRNP) H binds to the NRS G-rich elements and that hnRNP H is required for optimal U11 binding in vitro. It is further shown that hnRNP H (but not hnRNP F) can promote U11 binding and splicing from the NRS in vivo when tethered to the RNA as an MS2 fusion protein. Interestingly, 17% of the naturally occurring U12-dependent introns have at least two potential hnRNP H binding sites positioned similarly to the NRS. For two such introns from the SCN4A and P120 genes, we show that hnRNP H binds to each in a G tract-dependent manner, that G-tract mutations strongly reduce splicing of minigene RNA, and that tethered hnRNP H restores splicing to mutant RNA. In support of a role for hnRNP H in both splicing pathways, hnRNP H antibodies co precipitate U1 and U11 small nuclear ribonucleoproteins. These results indicate that hnRNP H is an auxiliary factor for U11 binding to the NRS and that, more generally, hnRNP H is a splicing factor for a subset of U12-dependent introns that harbor G-rich elements. PMID- 16308320 TI - Involvement of vertebrate Polkappa in translesion DNA synthesis across DNA monoalkylation damage. AB - DNA lesions that escape excision repair pathways can cause arrested DNA replication. This replication block can be processed by translesion DNA synthesis (TLS), which is carried out by a number of specialized DNA polymerases. A sequential lesion bypass model has been proposed; one of the lesion-specific polymerases inserts nucleotide(s) opposite the damaged template, followed by extension from the inserted nucleotide by the same or another polymerase. Polzeta and Polkappa have been proposed as candidates for executing the extension step in eukaryotic cells. We previously disrupted separately Rev3, the catalytic subunit of Polzeta, and Polkappa in chicken B lymphocyte DT40 cells. We found that each cell line showed significant UV sensitivity, implying that both contribute to UV radiation damage repair. In the present studies we generated REV3(-/-)POLK(/-) double knock-out cells to determine whether they participate in the same or different pathways. The double mutant was viable and proliferated with the same kinetics as parental REV3(-/-) cells. The cells showed the same sensitivity as REV3(-/-) cells to UV, ionizing radiation, and chemical cross-linking agents. In contrast, they were more sensitive than REV3(-/-) cells to monofunctional alkylating agents, even though POLK(/-) cells barely exhibited increased sensitivity to those. Moreover Polk-deficient mouse embryonic stem and fibroblast cells, both of which have previously been shown to be sensitive to UV radiation, also showed moderate sensitivity to methyl methanesulfonate, a monofunctional alkylating agent. These data imply that Polkappa has a function in TLS past alkylated base adducts as well as UV radiation DNA damage in vertebrates. PMID- 16308321 TI - Arginine/serine-rich protein interaction domain-dependent modulation of a tau exon 10 splicing enhancer: altered interactions and mechanisms for functionally antagonistic FTDP-17 mutations Delta280K AND N279K. AB - Tau exon 10 splicing is altered by autosomal dominant mutations that cause frontotemporal dementia with parkinsonism chromosome 17-type and by unknown mechanisms in other related neurodegenerative disorders. Identifying cis- and trans-regulators of tau exon 10 splicing is therefore crucial for understanding disease mechanisms. We previously identified several splicing enhancers and silencers within exon 10 and intron 10. Here, we show that splicing factors SF2/ASF, Tra2beta, and a 50-kDa nuclear protein bind in vitro to the polypurine enhancer at the 5' end of exon 10. Disease splicing mutations N279K and Delta280K disrupt the enhancer and alter associations with these factors. N279K targets robustly bind Tra2beta compared with the normal enhancer, which may explain why N279K enhances exon 10 splicing in vivo. In contrast, factor associations with Delta280K targets are nearly undetectable, explaining why Delta280K almost abolishes exon 10 splicing in vivo. Small interfering RNA-mediated suppression of endogenous SF2/ASF and Tra2beta significantly reduces exon 10 splicing. Exogenous SF2/ASF dramatically enhances normal exon 10 splicing and efficiently rescues the Delta280K splicing defect. Domain deletion analyses show that the C-terminal RS domains of SF2/ASF and Tra2beta are required for normal exon 10 splicing in vivo. In contrast to Tra2beta, the SF2/ASF RS domain remains essential in the presence of a strengthened enhancer or when either weak splice site is strengthened. The data suggest that SF2/ASF has both essential and regulatory roles, whereas Tra2beta has a supporting role in exon 10 splicing. PMID- 16308322 TI - Molecular structure of a 9-MDa icosahedral pyruvate dehydrogenase subcomplex containing the E2 and E3 enzymes using cryoelectron microscopy. AB - The pyruvate dehydrogenase multienzyme complexes are among the largest multifunctional catalytic machines in cells, catalyzing the production of acetyl CoA from pyruvate. We have previously reported the molecular architecture of an 11-MDa subcomplex comprising the 60-mer icosahedral dihydrolipoyl acetyltransferase (E2) decorated with 60 copies of the heterotetrameric (alpha(2)beta(2)) 153-kDa pyruvate decarboxylase (E1) from Bacillus stearothermophilus (Milne, J. L. S., Shi, D., Rosenthal, P. B., Sunshine, J. S., Domingo, G. J., Wu, X., Brooks, B. R., Perham, R. N., Henderson, R., and Subramaniam, S. (2002) EMBO J. 21, 5587-5598). An annular gap of approximately 90 A separates the acetyltransferase catalytic domains of the E2 from an outer shell formed of E1 tetramers. Using cryoelectron microscopy, we present here a three dimensional reconstruction of the E2 core decorated with 60 copies of the homodimeric 100-kDa dihydrolipoyl dehydrogenase (E3). The E2E3 complex has a similar annular gap of approximately 75 A between the inner icosahedral assembly of acetyltransferase domains and the outer shell of E3 homodimers. Automated fitting of the E3 coordinates into the map suggests excellent correspondence between the density of the outer shell map and the positions of the two best fitting orientations of E3. As in the case of E1 in the E1E2 complex, the central 2-fold axis of the E3 homodimer is roughly oriented along the periphery of the shell, making the active sites of the enzyme accessible from the annular gap between the E2 core and the outer shell. The similarities in architecture of the E1E2 and E2E3 complexes indicate fundamental similarities in the mechanism of active site coupling involved in the two key stages requiring motion of the swinging lipoyl domain across the annular gap, namely the synthesis of acetyl CoA and regeneration of the dithiolane ring of the lipoyl domain. PMID- 16308323 TI - Tricuspid valve endocarditis and septic pulmonary emboli illustrated by ECG-gated multi-slice CT of the chest. PMID- 16308324 TI - Noise burden and the risk of myocardial infarction. AB - AIMS: Chronic noise exposure is associated with adverse pathophysiological effects and may contribute to the progression of cardiovascular disease. We, therefore, determined the risk of noise for the incidence of myocardial infarction. METHODS AND RESULTS: In a case-control study, 4115 patients (3054 men, 56+/-9 years; 1061 women, 58+/-9 years) consecutively admitted to all 32 major hospitals in Berlin with confirmed diagnosis of acute myocardial infarction were enrolled from 1998 to 2001 in the Noise and Risk of Myocardial Infarction (NaRoMI) study. Controls were matched for gender, age, and hospital. In standardized interviews, information was obtained on environmental and work noise annoyance. The sound levels of environmental and work noise were assessed using traffic noise maps as proxy and international standards for workplaces, respectively. In multivariate logistic regression models, the adjusted odds ratios of noise variables were determined. There was a marginally increased risk of myocardial infarction associated with annoyance by environmental noise in women (adjusted odds ratio 1.47, 95% confidence interval 0.95-2.25, P=0.081) but not in men, and not associated with annoyance by work noise. Environmental sound levels were associated with increased risk in men and women (odds ratios 1.46, 1.02-2.09, P=0.040 and 3.36, 1.40-8.06, P=0.007) and work sound levels in men only (1.31, 1.01-1.70, P=0.045). CONCLUSION: Chronic noise burden is associated with the risk of myocardial infarction. The risk increase appears more closely associated with sound levels than with subjective annoyance. Further investigation of the gender-related risk of noise exposure may aid in improving prevention. PMID- 16308325 TI - Evidence-based common sense: the role of clinical history for the diagnosis of vasovagal syncope. PMID- 16308326 TI - Contrast ultrasound perfusion imaging of lower extremities in peripheral arterial disease: a novel diagnostic method. AB - AIMS: The purpose of this study was to establish contrast-enhanced ultrasound perfusion imaging (CUPI) of the lower extremities as a novel non-invasive diagnostic tool for patients with peripheral arterial disease (PAD). METHODS AND RESULTS: Ultrasound contrast agent (SonoVue) was injected into a peripheral vein of 16 control subjects and 16 PAD patients and its appearance in the calf muscle was detected by low-energy harmonic ultrasound. Analysis of the wash-in curves revealed that PAD patients had a significantly longer time to peak intensity (TTP), i.e. duration of maximum contrast perfusion [37 s (19-79 s) in control subjects vs. 56 s (32-104 s) in PAD patients at rest, age-adjusted P=0.002]. Exercise stress test of the calf muscle resulted in a decrease of the TTP, maintaining the significant difference in TTP between the groups [19 s (8-37 s) in control subjects vs. 32 s (18-48 s) in PAD patients after exercise, age adjusted P=0.004]. Neither ankle-brachial index and TTP nor age and TTP showed a significant correlation. CONCLUSION: CUPI reflects the regional blood circulation of the calf muscle. In this pilot study, PAD patients show a significantly longer TTP than control subjects. The clinical relevance of CUPI is topic of ongoing studies. PMID- 16308327 TI - Why do doctors issue sick notes? An experimental questionnaire study in primary care. AB - BACKGROUND: Issuing sick notes is one of the core tasks of General Practice and yet little research has explored how doctors decide whether or not to offer a sick note. AIM: To explore what factors influence this decision with a focus on the impact of type of problem (psychological versus physical), adverse family circumstances (present versus absent) and patient demand (asks for note versus does not ask). DESIGN: Experimental factorial design using questionnaire-based vignettes with eight scenarios which varied in terms of the three core factors. SETTING: East and West Sussex PCTs. OUTCOME MEASURES: Doctors' beliefs about the patient and their subsequent behaviour. PARTICIPANTS: Four hundred and eighty nine GPs completed a questionnaire asking them to rate one of eight hypothetical patients in terms of their beliefs about the patient and their hypothetical behaviour. RESULTS: The doctors rated the patient with the psychological problem as more ill, less work-shy, more unfit for work and described feeling more sympathy towards him compared with the patient with the physical problem. The presence of adverse family circumstances generated more sympathy and doctors considered this patient as less work-shy. A patient demand for a sick note had no effect on doctors' beliefs about the patient. In terms of doctors' behaviour, the doctors were more likely to give the patient with the psychological problem a sick note overall and because they felt he needed or deserved one, and more likely to give the patient with the physical problem a sick note in order to maintain a relationship with him. The decision to give a sick note was not influenced by either adverse family circumstances or patient demand. CONCLUSION: Doctors have more positive beliefs about patients with a psychological problem and are more likely to offer them a sick note. Issuing sick notes is unrelated to the patient's family circumstances or patient demand. PMID- 16308328 TI - What do stroke patients and their carers want from community services? AB - BACKGROUND: Previous research has focused on the longer term needs of 'new' stroke patients at fixed time intervals after the event, but neglected those of stroke patients who may have had the event many years earlier. OBJECTIVE: To identify the long-term support needs of patients with prevalent stroke, and their carers identified from practice stroke registers. DESIGN OF STUDY: Patients and their carers were invited to attend focus groups at the university, a nursing home or in the community. SETTING: Seven practices in South Birmingham. Adults (18+) with a validated record of stroke. METHODS: Focus groups were audio-taped and data analysed using a constant comparison method. RESULTS: Twenty-seven patients and six carers participated in the study. Three major themes emerged: emotional and psychological problems; lack of information available for patients and their families; the importance of Primary Care as the first point of contact for information or problems, even if these were non medical. CONCLUSIONS: Better methods of providing information for long-term survivors of stroke, and for addressing their emotional and psychological needs are required. Primary care could be a key setting for helping to provide more inclusive services for both patient and carer. PMID- 16308329 TI - Reciprocal epithelial-mesenchymal FGF signaling is required for cecal development. AB - Fibroblast growth factor (FGF) signaling mediates reciprocal mesenchymal epithelial cell interactions in the developing mouse lung and limb. In the gastrointestinal (GI) tract, FGF10 is expressed in the cecal mesenchyme and signals to an epithelial splice form of FGF receptor (FGFR) 2 to regulate epithelial budding. Here, we identify FGF9 as a reciprocal epithelial-mesenchymal signal required for cecal morphogenesis. Fgf9 null (Fgf9(-/-)) mouse embryos have agenesis of the embryonic cecum, lacking both mesenchymal expansion and an epithelial bud. In the cecal region of Fgf9(-/-) embryos, mesenchymal expression of Fgf10 and Bmp4 is notably absent, whereas the expression of epithelial markers, such as sonic hedgehog, is not affected. Using epithelial and whole explant cultures, we show that FGF9 signals to mesenchymal FGFRs and that FGF10 signals to epithelial FGFRs. Taken together, these data show that an epithelial FGF9 signal is necessary for the expansion of cecal mesenchyme and the expression of mesenchymal genes that are required for epithelial budding. Thus, these data add to our understanding of FGF-mediated reciprocal epithelial-mesenchymal signaling. PMID- 16308330 TI - Increasing Fgf4 expression in the mouse limb bud causes polysyndactyly and rescues the skeletal defects that result from loss of Fgf8 function. AB - A major function of the limb bud apical ectodermal ridge (AER) is to produce fibroblast growth factors (FGFs) that signal to the underlying mesenchyme. Previous studies have suggested that of the four FGF genes specifically expressed in the mouse AER, Fgf8 is unique not only in its expression pattern, but also because it is the only such FGF gene that causes limb skeletal abnormalities when individually inactivated. However, when both Fgf8 and Fgf4 are simultaneously inactivated in the AER, the limb does not develop. One possible explanation for these observations is that although both of these FGF family members contribute to limb development, Fgf8 has functions that Fgf4 cannot perform. To test this hypothesis, we used a novel method to substitute Fgf4 for Fgf8 expression in the developing limb bud by concomitantly activating a conditional Fgf4 gain-of function allele and inactivating an Fgf8 loss-of-function allele in the same cells via Cre-mediated recombination. Our data show that when Fgf4 is expressed in place of Fgf8, all of the skeletal defects caused by inactivation of Fgf8 are rescued, conclusively demonstrating that FGF4 can functionally replace FGF8 in limb skeletal development. We also show that the increase in FGF signaling that occurs when the Fgf4 gain-of-function allele is activated in a wild-type limb bud causes formation of a supernumerary posterior digit (postaxial polydactyly), as well as cutaneous syndactyly between all the digits. These data underscore the importance of controlling the level of FGF gene expression for normal limb development. PMID- 16308331 TI - MAP kinase subcellular localization controls both pattern and proliferation in the developing Drosophila wing. AB - Mitogen-activated protein kinases (MAPKs) phosphorylate target proteins in both the cytoplasm and nucleus, and a strong correlation exists between the subcellular localization of MAPK and resulting cellular responses. It was thought that MAPK phosphorylation was always followed by rapid nuclear translocation. However, we and others have found that MAPK phosphorylation is not always sufficient for nuclear translocation in vivo. In the developing Drosophila wing, MAPK-mediated signaling is required both for patterning and for cell proliferation, although the mechanism of this differential control is not fully understood. Here, we show that phosphorylated MAPK (pMAPK) is held in the cytoplasm in differentiating larval and pupal wing vein cells, and we show that this cytoplasmic hold is required for vein cell fate. At the same time, we show that MAPK does move into the nucleus of other wing cells where it promotes cell proliferation. We propose a novel Ras pathway bifurcation in Drosophila and our results suggest a mechanism by which MAPK phosphorylation can signal two different cellular outcomes (differentiation versus proliferation) based on the subcellular localization of MAPK. PMID- 16308332 TI - Vg 1 is an essential signaling molecule in Xenopus development. AB - Xenopus Vg 1, a transforming growth factor beta (Tgfbeta) family member, was one of the first maternally localized mRNAs identified in vertebrates. Its restriction to the vegetal pole of the egg made it the ideal candidate to be the mesoderm-inducing signal released by vegetal cells, but its function in vivo has never been resolved. We show that Vg 1 is essential for Xenopus embryonic development, and is required for mesoderm induction and for the expression of several key Bmp antagonists. Although the original Vg 1 transcript does not rescue Vg 1-depleted embryos, we report that a second allele is effective. This work resolves the mystery of Vg 1 function, and shows it to be an essential maternal regulator of embryonic patterning. PMID- 16308333 TI - Reproductive exile versus reproductive tourism. PMID- 16308334 TI - Relationship between urinary estrogen levels before conception and sex ratio at birth in a primate, the gray mouse lemur. PMID- 16308335 TI - Elevated MMP-12 protein levels in induced sputum from patients with COPD. AB - BACKGROUND: Several matrix metalloproteinases (MMPs) are involved in the pathogenesis of chronic obstructive pulmonary disease (COPD). In mice, MMP-12 plays a crucial role in the development of cigarette smoke induced emphysema. A study was undertaken to investigate the role of MMP-12 in the development of COPD in human smokers. METHODS: Induced sputum samples were collected from patients with stable COPD (n = 28), healthy smokers (n = 14), never smokers (n = 20), and former smokers (n = 14). MMP-12 protein levels in induced sputum were determined by ELISA and compared between the four study groups. MMP-12 enzymatic activity in induced sputum was evaluated by casein zymography and by cleaving of a fluorescence quenched substrate. RESULTS: Median (IQR) MMP-12 levels were significantly higher in COPD patients than in healthy smokers, never smokers, and former smokers (17.5 (7.1-42.1) v 6.7 (3.9-10.4) v 4.2 (2.4-11.3) v 6.1 (4.5-7.6) ng/ml, p = 0.0002). MMP-12 enzymatic activity was significantly higher in patients with COPD than in controls (4.11 (1.4-8.0) v 0.14 (0.1-0.2) microg/microl, p = 0.0002). CONCLUSION: MMP-12 is markedly increased in induced sputum from patients with stable COPD compared with controls, suggesting a role for MMP-12 in the development of COPD in smokers. PMID- 16308336 TI - Lung function decline in asthma: association with inhaled corticosteroids, smoking and sex. AB - BACKGROUND: Inhaled corticosteroids (ICS) provide short term benefits in asthma but the long term effects are still unknown. METHODS: 281 patients diagnosed with moderate to severe asthma in 1963-75 were re-examined in 1991-9. Information was collected on forced expiratory volume in 1 second (FEV(1)), bronchial hyperresponsiveness, atopy, smoking, use and dosage of oral and ICS. Patients were included in the analyses if they had at least three FEV(1) measurements during two consecutive years after the age of 30 and used ICS during follow up. RESULTS: Analyses were performed on 122 patients. During a median follow up period of 23 years, 71 men and 51 women had on average 37 and 40 individual FEV(1) measurements, respectively. Linear mixed effect models showed that men had a mean annual decline in FEV(1) of 20.6 ml/year less after ICS initiation than before (p = 0.011), and in women the decline in FEV(1) was 3.2 ml/year less (p = 0.73). In individuals with <5 pack years of smoking the decline in FEV(1) was 36.8 ml/year less after ICS institution in men (p = 0.0097) and 0.8 ml/year less in women (p = 0.94), the difference between the sexes being significant (p = 0.045). These effects were not observed in those with > or =5 pack years smoking. A higher daily dose of ICS was associated with a smaller decline in FEV(1) in men (p = 0.006), an effect not observed in women. CONCLUSION: Treatment with ICS in adult patients with moderate to severe asthma was associated with a reduction in the decline in FEV(1) over a 23 year follow up period in men who had smoked <5 pack years. This effect was dose dependent and was not present in women or in men with > or =5 pack years of smoking at follow up. The lack of effect of ICS on the decline in FEV(1) in women needs further study. PMID- 16308337 TI - Comprehensive multigene phylogenies of excavate protists reveal the evolutionary positions of "primitive" eukaryotes. AB - Many of the protists thought to represent the deepest branches on the eukaryotic tree are assigned to a loose assemblage called the "excavates." This includes the mitochondrion-lacking diplomonads and parabasalids (e.g., Giardia and Trichomonas) and the jakobids (e.g., Reclinomonas). We report the first multigene phylogenetic analyses to include a comprehensive sampling of excavate groups (six nuclear-encoded protein-coding genes, nine of the 10 recognized excavate groups). Excavates coalesce into three clades with relatively strong maximum likelihood bootstrap support. Only the phylogenetic position of Malawimonas is uncertain. Diplomonads, parabasalids, and the free-living amitochondriate protist Carpediemonas are closely related to each other. Two other amitochondriate excavates, oxymonads and Trimastix, form the second monophyletic group. The third group is comprised of Euglenozoa (e.g., trypanosomes), Heterolobosea, and jakobids. Unexpectedly, jakobids appear to be specifically related to Heterolobosea. This tree topology calls into question the concept of Discicristata as a supergroup of eukaryotes united by discoidal mitochondrial cristae and makes it implausible that jakobids represent an independent early diverging eukaryotic lineage. The close jakobids-Heterolobosea-Euglenozoa connection demands complex evolutionary scenarios to explain the transition between the presumed ancestral bacterial-type mitochondrial RNA polymerase found in jakobids and the phage-type protein in other eukaryotic lineages, including Euglenozoa and Heterolobosea. PMID- 16308338 TI - Patterns of protein evolution in Tetrahymena thermophila: implications for estimates of effective population size. AB - High levels of synonymous substitutions among alleles of the surface antigen SerH led to the hypothesis that Tetrahymena thermophila has a tremendously large effective population size, one that is greater than estimated for many prokaryotes (Lynch, M., and J. S. Conery. 2003. Science 302:1401-1404.). Here we show that SerH is unusual as there are substantially lower levels of synonymous variation at five additional loci (four nuclear and one mitochondrial) characterized from T. thermophila populations. Hence, the effective population size of T. thermophila, a model single-celled eukaryote, is lower and more consistent with estimates from other microbial eukaryotes. Moreover, reanalysis of SerH polymorphism data indicates that this protein evolves through a combination of vertical transmission of alleles and concerted evolution of repeat units within alleles. SerH may be under balancing selection due to a mechanism analogous to the maintenance of antigenic variation in vertebrate immune systems. Finally, the dual nature of ciliate genomes and particularly the amitotic divisions of processed macronuclear genomes may make it difficult to estimate accurately effective population size from synonymous polymorphisms. This is because selection and drift operate on processed chromosomes in macronuclei, where assortment of alleles, disruption of linkage groups, and recombination can alter the genetic landscape relative to more canonical eukaryotic genomes. PMID- 16308340 TI - The process of producing recommendations for rheumatic diseases: what is evidence? PMID- 16308339 TI - Methotrexate modulates the kinetics of adenosine in humans in vivo. AB - BACKGROUND: Animal studies suggest that the anti-inflammatory effect of methotrexate (MTX) is mediated by increased adenosine concentrations. OBJECTIVE: To assess the effect of MTX on the vasodilator effects of adenosine and the nucleoside uptake inhibitor, dipyridamole, in humans in vivo as a marker for changes in adenosine kinetics. METHODS: Ten patients with active arthritis were treated with MTX (15 mg/week). Measurements were performed before and after 12 weeks of treatment. At these time points, the activity of adenosine deaminase was measured in isolated lymphocytes, and forearm blood flow (FBF) was determined by venous occlusion plethysmography during administration of adenosine and dipyridamole into the brachial artery. RESULTS: The Vmax of adenosine deaminase in lymphocytes was reduced by MTX treatment (p<0.05). MTX significantly enhanced vasodilator response to adenosine (0.5 and 1.5 microg/min/dl of forearm tissue; mean (SE) FBF ratio increased from 1.2 (0.2) to 1.4 (0.2) and 2.2 (0.2) ml/dl/min, respectively, before and from 1.3 (0.1) to 1.8 (0.2) and 3.2 (0.5) ml/dl/min during MTX treatment; p<0.05). Also, dipyridamole-induced vasodilatation (30 and 100 microg/min/dl) was enhanced by MTX (FBF ratio increased from 1.2 (0.2) to 1.5 (0.3) and 1.8 (0.2), respectively, before and from 1.3 (0.1) to 1.8 (0.2) and 2.4 (0.4) during MTX treatment; p<0.05). CONCLUSIONS: MTX treatment inhibits deamination of adenosine and potentiates adenosine-induced vasodilatation. Also dipyridamole-induced vasodilatation is enhanced by MTX treatment, suggesting an increased extracellular formation of adenosine. These effects on the adenosine kinetics in humans may contribute to the therapeutic efficacy of MTX. PMID- 16308341 TI - Long term efficacy and safety of adalimumab plus methotrexate in patients with rheumatoid arthritis: ARMADA 4 year extended study. AB - OBJECTIVE: To evaluate the efficacy and safety of adalimumab plus methotrexate (MTX) given for up to 4 years in patients with active, longstanding rheumatoid arthritis. METHODS: Patients responding inadequately to MTX were entered into a 24 week, controlled study (ARMADA) with adalimumab plus MTX or placebo plus MTX, and some were enrolled in a subsequent open label extension. The efficacy and safety of treatment were evaluated. Additional analyses were made for those patients whose corticosteroid and/or MTX dosages were adjusted during the extension. RESULTS: Of 271 patients in the original ARMADA trial, 262 received at least one dose of adalimumab and were evaluated. At the time of analysis, 162/262 (62%) patients had remained in the study and received treatment for a mean of 3.4 years. Withdrawals were for lack of efficacy (8%), adverse events (12%), and other reasons (18%). In 147 patients who completed 4 years' treatment, efficacy achieved at 6 months was maintained. At 4 years, 78%, 57%, and 31% had achieved ACR20/50/70; 43% achieved clinical remission (DAS28 <2.6); and 22% had no physical function abnormalities (HAQ = 0). Results were similar for 196 patients who received treatment for 2-4 years. Efficacy was maintained in many patients when dosages were decreased (corticosteroids (51/81 (63%) patients), MTX (92/217 (42%)), or both (25/217 (12%))). Serious adverse events were comparable during open label treatment and the controlled phase. Serious infections occurring during open label treatment and the blinded period were similar (2.03 v 2.30 events per 100 patient-years, respectively). CONCLUSIONS: Adalimumab plus MTX sustained clinical response and remission in patients with RA during 4 years. The safety profile during the first 6 months was similar to that after 4 years' follow up. Reduction of corticosteroid and/or MTX dosages did not adversely affect long term efficacy. PMID- 16308342 TI - Relative impact of radiographic osteoarthritis and pain on quadriceps strength, proprioception, static postural sway and lower limb function. AB - OBJECTIVE: To investigate the relative impact of radiographic osteoarthritis (ROA) and current knee pain on lower limb physical function, quadriceps strength, knee joint proprioception, and postural sway. METHODS: Using a 2x2 factorial design, 142 community derived subjects aged over 45 were divided into four subgroups based on the presence or absence of ROA (Kellgren & Lawrence>grade 2) and knee pain (as assessed by NHANES questions and a 100 mm visual analogue scale). Maximum isometric contraction of the quadriceps, knee joint proprioceptive acuity, static postural sway, and WOMAC index (both whole and function subscale) were assessed in all subjects. RESULTS: Compared with normal subjects, reported disability was greater for all other subgroups (p<0.01). Subjects with both ROA and knee pain reported the greatest disability, and those with knee pain only had greater disability than those with ROA only. Quadriceps weakness was observed in all groups compared with normal subjects (p<0.01), though they were no significant intergroup differences. Subjects with knee pain had a greater sway area than those without (p<0.05) but the presence of ROA was not associated with increased postural sway. No differences in proprioceptive acuity were observed between groups. CONCLUSIONS: The presence of knee pain has a negative association with quadriceps strength, postural sway, and disability compared with ROA. However, the presence of pain-free ROA has a significant negative influence on relative quadriceps strength and reported disability. PMID- 16308343 TI - "Periodic fever" without fever: two cases of non-febrile TRAPS with mutations in the TNFRSF1A gene presenting with episodes of inflammation or monosymptomatic amyloidosis. AB - BACKGROUND: Tumour necrosis factor (TNF) receptor associated periodic syndrome (TRAPS) is caused by dominant mutations in the TNFRSF1A gene. In typical cases TRAPS begins early in childhood and is characterised by high and remittent fever over a period of 1-4 weeks or longer, accompanied by systemic and local inflammation. CASE REPORTS: Patient 1 presented with recurrent episodes of weakness, migrating myalgias, arthralgias, exanthema, and chest pain lasting for 1-4 weeks, but without any fever over an initial period of 4 years at least. Diagnosis of TRAPS was confirmed by the heterozygous mutation Y20H in TNFRSF1A. Patient 2, a 23 year old woman never had any symptoms indicative of TRAPS. Genetic evaluation of all members of her family with a TRAPS index patient disclosed the T50M mutation in TNFRSF1A. A medical check up showed proteinuria, and renal biopsy disclosed AA amyloidosis. CONCLUSIONS: TRAPS associated mutations can induce considerable inflammation that is not necessarily accompanied by fever. Even monosymptomatic severe amyloidosis can occur in these patients. Genetic counselling and appropriate management to prevent or mitigate amyloidosis may be necessary. PMID- 16308344 TI - Low plasma protein nitrotyrosine levels distinguish primary Raynaud's phenomenon from scleroderma. AB - OBJECTIVE: To investigate the hypothesis that increased formation of reactive nitrogen species may contribute to the vascular pathology that develops in patients with connective tissue disease such as scleroderma. PATIENTS AND METHODS: The level of protein-bound nitrotyrosine in plasma was measured by stable isotope dilution gas chromatography/negative ion chemical ionisation mass spectrometry in 11 patients with primary Raynaud's phenomenon, 37 with scleroderma, 13 with chronic renal impairment, and in 23 healthy controls. RESULTS: Plasma protein-bound nitrotyrosine was markedly decreased in patients with primary Raynaud's phenomenon (mean (SEM) 0.60 (0.06) ng/mg dry protein) compared with patients with scleroderma (1.78 (0.21) ng/mg protein), chronic renal impairment (1.42 (0.17) ng/mg protein) or healthy controls (1.63+/-0.15 ng/mg protein, ANOVA p<0.001). CONCLUSION: These data suggest that there is decreased nitration of plasma proteins, or increased degradation of nitrated proteins from the circulation of patients with primary but not secondary Raynaud's phenomenon. PMID- 16308345 TI - Novel two-rhythm generator theory of breathing in mammals. PMID- 16308346 TI - A role for 5,6-epoxyeicosatrienoic acid in calcium entry by de novo conformational coupling in human platelets. AB - A major pathway for Ca(2+) entry in non-excitable cells is activated following depletion of intracellular Ca(2+) stores. A de novo conformational coupling between elements in the plasma membrane (PM) and Ca(2+) stores has been proposed as the most likely mechanism to activate this capacitative Ca(2+) entry (CCE) in several cell types, including platelets. Here we report that a cytochrome P450 metabolite, 5,6-EET, might be a component of the de novo conformational coupling in human platelets. In these cells, 5,6-EET induces divalent cation entry without having any detectable effect on Ca(2+) store depletion. 5,6-EET-induced Ca(2+) entry was sensitive to the CCE blockers 2-APB, lanthanum, SKF-96365 and nickel and impaired by incubation with anti-hTRPC1 antibody. Ca(2+) entry stimulated by low concentrations of thapsigargin, which selectively depletes the dense tubular system and induces EET production, was impaired by the cytochrome P450 inhibitor 17-ODYA, which has no effect on CCE mediated by depletion of the acidic stores using 2,5-di-(tert-butyl)-1,4-hydroquinone. We have found that 5,6-EET-induced Ca(2+) entry requires basal levels of H(2)O(2), which might maintain a redox state favourable for this event. Finally, our results indicate that 5,6-EET induces the activation of tyrosine kinase proteins and the reorganization of the actin cytoskeleton, which might provide a support for the transport of portions of the Ca(2+) store towards the PM to facilitate de novo coupling between IP(3)R type II and hTRPC1 detected by coimmunoprecipitation. We propose that the involvement of 5,6-EET in TG-induced coupling between IP(3)R type II and hTRPC1 and subsequently CCE is compatible with the de novo conformational coupling in human platelets. PMID- 16308347 TI - Interaction of KCNE subunits with the KCNQ1 K+ channel pore. AB - KCNQ1 alpha subunits form functionally distinct potassium channels by coassembling with KCNE ancillary subunits MinK and MiRP2. MinK-KCNQ1 channels generate the slowly activating, voltage-dependent cardiac IKs current. MiRP2 KCNQ1 channels form a constitutively active current in the colon. The structural basis for these contrasting channel properties, and the mechanisms of alpha subunit modulation by KCNE subunits, are not fully understood. Here, scanning mutagenesis located a tryptophan-tolerant region at positions 338-340 within the KCNQ1 pore-lining S6 domain, suggesting an exposed region possibly amenable to interaction with transmembrane ancillary subunits. This hypothesis was tested using concomitant mutagenesis in KCNQ1 and in the membrane-localized 'activation triplet' regions of MinK and MiRP2 to identify pairs of residues that interact to control KCNQ1 activation. Three pairs of mutations exerted dramatic effects, ablating channel function or either removing or restoring control of KCNQ1 activation. The results place KCNE subunits close to the KCNQ1 pore, indicating interaction of MiRP2-72 with KCNQ1-338; and MinK-59,58 with KCNQ1-339, 340. These data are consistent either with perturbation of the S6 domain by MinK or MiRP2, dissimilar positioning of MinK and MiRP2 within the channel complex, or both. Further, the results suggest specifically that two of the interactions, MiRP2 72/KCNQ1-338 and MinK-58/KCNQ1-340, are required for the contrasting gating effects of MinK and MiRP2. PMID- 16308348 TI - Single-unit analysis of sympathetic nervous discharges in patients with panic disorder. AB - Patients with panic disorder are at increased cardiac risk. While the mechanisms responsible remain unknown, activation of the sympathetic nervous system may be implicated. Using isotope dilution methodology, investigations of whole-body and regional sympathetic nervous activity have failed to show any differences between patients with panic disorder and healthy subjects. Using direct recording of single unit efferent sympathetic vasoconstrictor nerve activity by microneurography we examined sympathetic nervous function in patients with panic disorder more precisely than previously reported. The activity of multiunit and single unit vasoconstrictor sympathetic nerves was recorded at rest at the level of the peroneal nerve in 10 patients diagnosed with panic disorder and in nine matched healthy volunteers. Multiunit sympathetic activity was not different between the two groups (26+/-3 bursts min-1 in patients with panic disorder and 28+/-3 bursts min-1 in controls). The firing frequency of single unit vasoconstrictor neurones was also similar between the two groups (0.38+/-0.09 versus 0.22+/-0.03 Hz). However, the probability of firing during a sympathetic burst was higher in patients with panic disorder compared with healthy controls (45+/-5%versus 32+/-3%, P<0.05). When only the neural bursts during which the vasoconstrictor neurone was active were considered, we found that in patients with panic disorder the neurones tended to fire more often in a 'multiple spike' pattern than in the controls (i.e. the probability of the neurone firing twice was 25+/-3% in patients with panic disorder compared with 14+/-3% in controls). Quantification from single vasoconstrictor unit recording provides evidence of a disturbed sympathetic firing pattern in patients with panic disorder. PMID- 16308349 TI - Simulation of Ca2+ persistent inward currents in spinal motoneurones: mode of activation and integration of synaptic inputs. AB - The goal of this study was to investigate the nature of activation of the dendritic calcium persistent inward current (Ca(2+) PIC) and its contribution to the enhancement and summation of synaptic inputs in spinal motoneurones. A compartmental cable model of a cat alpha-motoneurone was developed comprising the realistic dendritic distribution of Ia-afferent synapses and low-voltage activated L-type calcium (Ca(v)1.3) channels distributed over the dendrites in a manner that was previously shown to match a wide set of experimental measurements. The level of synaptic activation was systematically increased and the resulting firing rate, somatic and dendritic membrane potentials, dendritic Ca(v)1.3 channel conductance, and dendritic Ca(2+) PIC were measured. Our simulation results suggest that during cell firing the dendritic Ca(2+) PIC is not activated in an all-or-none manner. Instead, it is initially activated in a graded manner with increasing synaptic input until it reaches its full activation level, after which additional increases in synaptic input result in minimal changes in the Ca(2+) PIC (PIC saturated). The range of graded activation of Ca(2+) PIC occurs when the cell is recruited and causes a steep increase in the firing frequency as the synaptic current is increased, coinciding with the secondary range of the synaptic frequency-current (F-I) relationship. Once the Ca(2+) PIC is saturated the slope of the F-I relationship is reduced, corresponding to the tertiary range of firing. When the post-spike afterhyperpolarization (AHP) is blocked, either directly by blocking the calcium activated potassium channels, or indirectly by blocking the sodium spikes, the PIC is activated in an all-or-none manner with increasing synaptic input. Thus, the AHP serves to limit the depolarization of the cell during firing and enables graded, rather than all-or-none, activation of the Ca(2+) PIC. The graded activation of the Ca(2+) PIC with increasing synaptic input results in a graded (linear) enhancement and linear summation of synaptic inputs. In contrast, the saturated Ca(2+) PIC enhances synaptic inputs by a constant amount (constant current), and leads to less-than linear summation of multiple synaptic inputs. These model predictions improve our understanding of the mode of activation of the dendritic Ca(2+) PIC and its role in the enhancement and integration of synaptic inputs. PMID- 16308350 TI - Cholinergic control of excitability of spinal motoneurones in the salamander. AB - The cholinergic modulation of the electrical properties of spinal motoneurones was investigated in vitro, with the use of the whole-cell patch-clamp recording technique in lumbar spinal cord slices from juvenile urodeles (Pleurodeles waltlii). Bath application of acetylcholine (20 microM) with eserine (20 microM) induced an increase in the resting membrane potential, a decrease of the input resistance, a decrease of the action potential amplitude, and a reduction of the medium afterhyperpolarization (mAHP) that followed each action potential. Moreover, the firing rate of motoneurones during a depolarizing current pulse and the slope of their stimulus current-spike frequency relation were increased. All of these effects were mimicked by extracellular application of muscarine (20 microM), and blocked by application of the muscarinic receptor antagonist atropine (0.1-1 microM). They were not observed during bath application of nicotine (10 microM). These results suggest that the cholinergic modulation of spinal motoneurone excitability was mediated by activation of muscarinic receptors. Our results further show that the muscarinic action primarily resulted from a reduction of the Ca2+-activated K+ current responsible for the mAHP, an inhibition of the hyperpolarization-activated cation current, Ih, and an enhancement of the inward rectifying K+ current, I(Kir). We conclude that cholinergic modulation can contribute significantly to the production of motor behaviour by altering several ionic conductances responsible for the repetitive discharge of motoneurones. PMID- 16308351 TI - Thermogenic responsiveness to beta-adrenergic stimulation is augmented in exercising versus sedentary adults: role of oxidative stress. AB - Beta-adrenergic receptor (beta-AR) modulation of resting and postprandial energy expenditure (EE) is augmented in regularly exercising compared with sedentary adults, but the underlying physiological mechanisms are unknown. Differences in thermogenic responsiveness to beta-AR stimulation, perhaps secondary to reactive oxygen species (ROS) bioactivity, may be involved. To determine habitual exercise related differences in beta-AR thermogenic responsiveness and the possible influence of ROS, we measured the percentage increase in EE (DeltaEE%; indirect calorimetry, ventilated hood method) above resting EE in response to non-specific beta-AR stimulation (intravenous isoproterenol (isoprenaline): 6, 12 and 24 ng (kg fat-free mass)-1 min-1) in 25 sedentary (11 males; 51+/-4 years; body mass index 25.0+/-0.8 kg m-2, maximal oxygen uptake 29+/-1 ml kg-1 min-1 (mean+/ s.e.m.)) and 14 habitually aerobic exercising (9 males, 46+/-6 years, 23.1+/-0.7 kg m-2, 44+/-3 ml kg-1 min-1) healthy adults under normal (control) conditions and during acute intravenous administration of a potent antioxidant, ascorbic acid (vitamin C; 0.04 g (kg fat-free mass)-1). DeltaEE% was greater (P=0.02) in the habitually exercising (8.6+/-1.2, 12.9+/-1.2, 20.0+/-1.4) versus sedentary (6.3+/-0.7, 10.4+/-0.8, 16.0+/-1.0) adults. Ascorbic acid increased (P=0.01) DeltaEE% only in the sedentary adults (to 9.5+/-0.9, 12.4+/-0.7, 18.5+/-0.8), abolishing baseline group differences. DeltaEE% was not related to the amount of body fat, sex, or any other baseline characteristic. Thermogenic responsiveness to beta-AR stimulation is augmented in habitually exercising adults. The mechanism is ascorbic acid dependent, suggesting that it may be linked to decreased ROS bioactivity. Our findings advance a novel mechanism by which habitual physical activity may modulate EE in humans, with potential implications for energy balance and body weight control. PMID- 16308352 TI - Neuropsin is essential for early processes of memory acquisition and Schaffer collateral long-term potentiation in adult mouse hippocampus in vivo. AB - Long-term potentiation (LTP) is thought to be particularly important in the acquisition of hippocampus-associated memory, in part because it develops quickly and persists for indefinite periods. Extracellular proteolysis has been hypothesized to contribute to LTP by modifying adhesive relations of synapses and thus the morphology of excitatory synapses. Here we report that neuropsin (NP), an extracellular serine protease, is critically involved in the formation of both the potentiation effect and hippocampus-dependent forms of memory. NP-knockout mice were significantly impaired in the Morris water maze and Y-mazes and failed to exhibit early phase LTP induced by a single tetanus. Potentiation was also impaired or completely blocked by in vivo application of a specific inhibitor or a neutralizing monoclonal antibody for NP. Intriguingly, recombinant (r-) NP alone, without tetanic stimulation, elicited either long-lasting potentiation or depression, depending on the applied dose. The r-NP-elicited potentiation was occluded by prior induction of LTP, while theta-burst-elicited LTP was occluded by application of r-NP alone, suggesting that the two forms of plasticity have a common signalling pathway. r-NP-elicited potentiation and depression increased phosphorylation at different sites on the GluR1 subunit of the AMPA receptor that had previously been associated with LTP or long-term depression. Thus, we conclude that NP is necessary for establishment of LTP and has a significant role in memory acquisition. PMID- 16308353 TI - Neurones associated with saccade metrics in the monkey central mesencephalic reticular formation. AB - Neurones in the central mesencephalic reticular formation (cMRF) begin to discharge prior to saccades. These long lead burst neurones interact with major oculomotor centres including the superior colliculus (SC) and the paramedian pontine reticular formation (PPRF). Three different functions have been proposed for neurones in the cMRF: (1) to carry eye velocity signals that provide efference copy information to the SC (feedback), (2) to provide duration signals from the omnipause neurones to the SC (feedback), or (3) to participate in the transformation from the spatial encoding of a target selection signal in the SC into the temporal pattern of discharge used to drive the excitatory burst neurones in the pons (feed-forward). According to each respective proposal, specific predictions about cMRF neuronal discharge have been formulated. Individual neurones should: (1) encode instantaneous eye velocity, (2) burst specifically in relation to saccade duration but not to other saccade metrics, or (3) have a spectrum of weak to strong correlations to saccade dynamics. To determine if cMRF neurones could subserve these multiple oculomotor roles, we examined neuronal activity in relation to a variety of saccade metrics including amplitude, velocity and duration. We found separate groups of cMRF neurones that have the characteristics predicted by each of the proposed models. We also identified a number of subgroups for which no specific model prediction had previously been established. We found that we could accurately predict the neuronal firing pattern during one type of saccade behaviour (visually guided) using the activity during an alternative behaviour with different saccade metrics (memory guided saccades). We suggest that this evidence of a close relationship of cMRF neuronal discharge to individual saccade metrics supports the hypothesis that the cMRF participates in multiple saccade control pathways carrying saccade amplitude, velocity and duration information within the brainstem. PMID- 16308354 TI - Clinical implications of continued alcohol consumption after diagnosis of upper aerodigestive tract cancer. AB - AIMS: To examine clinical implications of findings on the relationship between continued alcohol consumption in patients after diagnosis and treatment of upper aerodigestive tract cancer. METHODS: Clinical research literature on the prevalence and effects of alcohol consumption after oral cancer diagnosis was reviewed. Since limited research is currently available on this important clinical topic, all published studies were considered regardless of size and methodology. RESULTS: Between 34 and 57% of oral cancer patients continue to drink alcohol after cancer diagnosis. Continued drinking increases complications from surgery, increases the likelihood of recurrent cancer, and reduces disease specific survival. Older patients and those with a longer and heavier drinking pattern prior to diagnosis are more likely to continue drinking after diagnosis. CONCLUSIONS: Findings indicate that routine alcohol screening of newly diagnosed oral cancer patients as well as brief intervention and/or treatment referral is warranted. Monitoring of alcohol consumption for the first year after diagnosis and treatment is recommended. PMID- 16308355 TI - Bioinformatic analysis of exon repetition, exon scrambling and trans-splicing in humans. AB - MOTIVATION: Using bioinformatic approaches we aimed to characterize poorly understood abnormalities in splicing known as exon scrambling, exon repetition and trans-splicing. RESULTS: We developed a software package that allows large scale comparison of all human expressed sequence tags (EST) sequences to the entire set of human gene sequences. Among 5,992,495 EST sequences, 401 cases of exon repetition and 416 cases of exon scrambling were found. The vast majority of identified ESTs contain fragments rather than full-length repeated or scrambled exons. Their structures suggest that the scrambled or repeated exon fragments may have arisen in the process of cDNA cloning and not from splicing abnormalities. Nevertheless, we found 11 cases of full-length exon repetition showing that this phenomenon is real yet very rare. In searching for examples of trans-splicing, we looked only at reproducible events where at least two independent ESTs represent the same putative trans-splicing event. We found 15 ESTs representing five types of putative trans-splicing. However, all 15 cases were derived from human malignant tissues and could have resulted from genomic rearrangements. Our results provide support for a very rare but physiological occurrence of exon repetition, but suggest that apparent exon scrambling and trans-splicing result, respectively, from in vitro artifact and gene-level abnormalities. AVAILABILITY: Exon-Intron Database (EID) is available at http://www.meduohio.edu/bioinfo/eid. Programs are available at http://www.meduohio.edu/bioinfo/software.html. The Laboratory website is available at http://www.meduohio.edu/medicine/fedorov SUPPLEMENTARY INFORMATION: Supplementary file is available at http://www.meduohio.edu/bioinfo/software.html. PMID- 16308356 TI - Veterinary public health. PMID- 16308357 TI - A walk on the wild side--emerging wildlife diseases. PMID- 16308358 TI - UK government collaborations to manage threats to animal and human health. PMID- 16308359 TI - Foodborne zoonoses. PMID- 16308360 TI - Resistance to antimicrobials in humans and animals. PMID- 16308361 TI - What could Dr Finlay and Mr Herriot learn from each other? PMID- 16308362 TI - Human health and nature conservation. PMID- 16308364 TI - Vaccinating poultry against avian flu is contributing to spread. PMID- 16308365 TI - Rates of sexually transmitted infections continue to rise. PMID- 16308366 TI - Number of people infected with HIV worldwide reaches 40m. PMID- 16308367 TI - Drug industry's new code criticised for lacking teeth. PMID- 16308368 TI - Medicare's new prescription drug plan causes confusion. PMID- 16308370 TI - Anaesthetist could face prosecution after GMC strikes her off. PMID- 16308374 TI - Do vets and doctors face similar ethical challenges? PMID- 16308375 TI - Pregnant women in poor countries face domestic violence. PMID- 16308377 TI - Smoking ban in prisons would lead to more assaults on staff. PMID- 16308379 TI - UK's regulatory framework gives it an advantage in stem cell research. PMID- 16308380 TI - UK guidelines urge improved recognition of obsessive-compulsive disorder. PMID- 16308381 TI - Harnessing Mother Nature against your fellow humans. PMID- 16308382 TI - Randomised controlled trial of animal facilitated therapy with dolphins in the treatment of depression. AB - OBJECTIVE: To evaluate the effectiveness of animal facilitated therapy with dolphins, controlling for the influence of the natural setting, in the treatment of mild to moderate depression and in the context of the biophilia hypothesis. SETTING: The study was carried out in Honduras, and recruitment took place in the United States and Honduras. DESIGN: Single blind, randomised, controlled trial. PARTICIPANTS: Outpatients, recruited through announcements on the internet, radio, newspapers, and hospitals. RESULTS: Of the 30 patients randomly assigned to the two groups of treatment, two dropped out of the treatment group after the first week and three withdrew their consent in the control group after they had been randomly allocated. For the participants who completed the study, the mean severity of the depressive symptoms was more reduced in the treatment group than in the control group (Hamilton rating scale for depression, P = 0.002; Beck depression inventory, P = 0.006). For the sample analysed by modified intention to treat and last observation carried forward, the mean differences for the Hamilton and Beck scores between the two groups was highly significant (P = 0.007 and P = 0.012, respectively). CONCLUSIONS: The therapy was effective in alleviating symptoms of depression after two weeks of treatment. Animal facilitated therapy with dolphins is an effective treatment for mild to moderate depression, which is based on a holistic approach, through interaction with animals in nature. PMID- 16308383 TI - Sleeping sickness in Uganda: a thin line between two fatal diseases. AB - OBJECTIVE: To determine, through the use of molecular diagnostic tools, whether the two species of parasite that cause human African trypanosomiasis have become sympatric. DESIGN: Blood sampling of all available patients between June 2001 and June 2005 in central Uganda and between July and September 2003 in northwest Uganda and analysis of subcounty sleeping sickness records in Uganda between 1985 and 2005. SETTING: Sleeping sickness treatment centres in central and northwest Uganda and in south Sudan. PARTICIPANTS: Patients presenting at the treatment centres and diagnosed as having sleeping sickness. MAIN OUTCOME MEASURE: Classification of parasites from patients from each disease focus as either Trypanosoma brucei rhodesiense (acute form) or T b gambiense (chronic form). RESULTS: Blood from 231 patients with sleeping sickness in central Uganda and from 91 patients with sleeping sickness in northwest Uganda and south Sudan were screened for T b rhodesiense (detection of SRA gene) and T b gambiense (detection of TgsGP gene). All samples from central Uganda were classified as T b rhodesiense, and all samples from northwest Uganda and south Sudan were identified as T b gambiense. CONCLUSIONS: The two focuses of human African trypanosomiasis remain discrete, but the area of Uganda affected by the acute form of human sleeping sickness has increased 2.5-fold since 1985, spreading to three new districts within the past five years through movement of infected livestock. Without preventive action targeted at the livestock reservoir of this zoonotic disease, it is likely that the two disease focuses will converge. This will have a major impact on diagnosis and treatment of this neglected disease. Real time monitoring is recommended, using molecular diagnostic tools (at a regional surveillance centre, for example) targeted at both livestock and human patients. PMID- 16308384 TI - Sleeping sickness--a growing problem? PMID- 16308385 TI - Treatment of bites by adders and exotic venomous snakes. PMID- 16308386 TI - Horses for courses. Comparative gastroenterology: common ground and collaborative potential. PMID- 16308387 TI - Pet ownership and human health: a brief review of evidence and issues. PMID- 16308388 TI - Pets--pleasures and problems. PMID- 16308389 TI - Early qualitative risk assessment of the emerging zoonotic potential of animal diseases. PMID- 16308390 TI - Zoonotic potential of emerging animal diseases. PMID- 16308391 TI - The human story. PMID- 16308392 TI - The animal story. PMID- 16308393 TI - Synergy between public health and veterinary services to deliver human and animal health interventions in rural low income settings. PMID- 16308394 TI - Human and animal health: strengthening the links: new foundation will help bring two disciplines together. PMID- 16308395 TI - Human and animal health: strengthening the links: animal and human case for reforming current food policies. PMID- 16308396 TI - Classification of stillbirth: cause, condition, or mechanism? PMID- 16308397 TI - Classification of stillbirth: reclassification obscures things. PMID- 16308398 TI - Classification of stillbirth: classification is not explanation. PMID- 16308399 TI - Human and animal health: strengthening the links: preventing dog bites. PMID- 16308400 TI - Human and animal health: strengthening the links: combined courses for vets and doctors? PMID- 16308401 TI - Reassessing strategies for improving health: strategies should include nutrition. PMID- 16308402 TI - Including care home residents in clinical research is fraught. PMID- 16308403 TI - Transitional care for elderly people: intermediate care can be safe and reduce hospital use, but is it and does it? PMID- 16308404 TI - Transitional care for elderly people: at what cost? PMID- 16308405 TI - Reassessing strategies for improving health: strategies in West and Central Africa need to be revised. PMID- 16308406 TI - Criminal proceedings will hamper calls for open culture. PMID- 16308407 TI - Invest in people, not restructuring. PMID- 16308408 TI - Rates of exception reporting need to be addressed. PMID- 16308409 TI - How protective is breast feeding against diarrhoeal disease in infants in 1990s England? A case-control study. AB - AIMS: To assess the effect of several measures of infant feeding on diarrhoeal disease, and whether these effects vary according to markers of social deprivation. METHODS: Case-control study of diarrhoeal disease cases presenting to 34 general practices in England. Controls were stratified on age group, area deprivation index for the practice, and whether or not the practice was in London. Data were available on 304 infants (167 cases and 137 controls). RESULTS: After adjustment for confounders, breast feeding was associated with significantly less diarrhoeal disease. Associations were striking even in infants aged > or = 6 months. They did not vary by social class, but were greater in those living in rented council accommodation and in more crowded households. The effect of receiving no breast milk was stronger in more deprived areas than in less deprived areas. The effect of not receiving exclusive breast milk was stronger in more deprived areas than in less deprived areas. In formula fed infants, there was significantly more diarrhoeal disease in those not sterilising bottles/teats with steam or chemicals. The protective effect of breast feeding did not persist beyond two months after breast feeding had stopped. CONCLUSIONS: Breast feeding protects against diarrhoeal disease in infants in England although the degree of protection may vary across infants and wear off after breast feeding cessation. Education about the benefits of breast feeding and the risks of inadequate sterilisation should be targeted at carers in deprived areas or households. PMID- 16308410 TI - Paediatric emergency department anaphylaxis: different patterns from adults. AB - BACKGROUND AND AIMS: Data on acute paediatric anaphylaxis presentations to the emergency department (ED) are limited. All allergic presentations to one Australian paediatric ED were studied to determine epidemiological, clinical, and outcome data. METHODS: Retrospective, case based study of patients under 16 years attending one metropolitan, paediatric teaching hospital ED in Australia over three years. The medical records of patients presenting with generalised allergic reactions and anaphylaxis satisfying relevant ICD-9-CM diagnostic codes were studied. The incidence, age, sex ratio, co-morbidities, likely aetiology, clinical features, management, and disposal were determined. RESULTS: A total of 526 children with generalised allergic reactions, and 57 with anaphylaxis were included in the study. This represented incidences of 9.3:1000 ED presentations for generalised allergic reactions and 1:1000 for anaphylaxis. There were no fatalities. In anaphylaxis cases, a cause was recognised in 68.4%. Cutaneous features were present in 82.5%. A past history of asthma was reported in 36.8%. Adrenaline was used in 39.3% of severe anaphylaxis cases. The ED alone definitively cared for 97.8% of all patients. Follow up was inadequate in cases of anaphylaxis. CONCLUSIONS: This is the first reported incidence figure for paediatric anaphylaxis ED presentations in Australia, and is less than that reported in adults in the same local population. However, the incidence of generalised allergic reactions of 9.3:1000 was greater than in the adults. Virtually all paediatric allergic cases may be managed in the ED alone, provided that the importance of specialist follow up, particularly for severe anaphylaxis, is recognised. PMID- 16308411 TI - A randomised study of "wet wraps" versus conventional treatment for atopic eczema. AB - AIMS: To compare two treatments in common usage for children with atopic eczema: "wet wrap" bandages versus conventional topically applied ointments. METHODS: A total of 50 children (age 4-27 months) with moderate to severe eczema were randomised to one or other treatment. Two research nurses supervised the study. The first gave advice and support, and the second, blinded to the treatment modality being used, scored the change in eczema severity over a period of four weeks using the SCORAD eczema severity scale. RESULTS: Both treatments gave a benefit in overall SCORAD scores (mean change for wet wrap group was 53 to 24; for the conventional group, 41 to 17). There was no significant difference between the two groups in terms of overall improvement at four weeks or in the timescale of improvements. The amount of topical of topical steroid used was similar in both groups. The wet wrap group suffered significantly more skin infections requiring antibiotics. Carers reported that wet wraps were less easy to apply than conventional treatment. PMID- 16308412 TI - Socioeconomic status and childhood leukaemia: a review. AB - BACKGROUND: A long-held view links higher socioeconomic status (SES) to higher rates of childhood leukaemia. Some recent studies exhibit associations in the opposite direction. METHODS: We reviewed journal literature through August 2002 for associations between childhood leukaemia and socioeconomic measures. We determined the direction of each association and its P-value. We described the results with regard to study design, calendar period, geographic locale, and level of the socioeconomic measures (individual or ecological). For measures with sufficient number of results, we computed summary P-values across studies. RESULTS: Case-control studies conducted in North America since 1980 have involved subject interviews or self-administered questionnaires and have consistently reported inverse (negative) associations of childhood leukaemia with individual level measures of family income, mother's education, and father's education. In contrast, associations have been consistently positive with father's occupational class in record-based case-control studies and with average occupational class in ecological studies. CONCLUSIONS: Connections of SES measures to childhood leukaemia are likely to vary with place and time. Validation studies are needed to estimate SES-related selection and participation in case-control studies. Because different socioeconomic measures (such as income and education) and individual-level and ecological-level measures may represent different risk factors, we advise researchers to report these measures separately rather than in summary indices of social class. PMID- 16308414 TI - Can we ablate permanent atrial fibrillation? AB - There is growing evidence of the benefit of ablation in the treatment of drug refractory atrial fibrillation. PMID- 16308413 TI - Exercise effects on cardiac size and left ventricular diastolic function: relationships to changes in fitness, fatness, blood pressure and insulin resistance. AB - OBJECTIVES: To determine exercise training effects on cardiac size and left ventricular (LV) diastolic function and relationships of exercise induced changes in physiological and body composition parameters with cardiac parameters. DESIGN: Prospective, randomised controlled trial. SUBJECTS: Men and women (63.6 (5.7) years, body mass index 29.5 (4.4) kg/m(2)) with untreated hypertension (systolic blood pressure (BP) 130-159 or diastolic BP 85-99 mm Hg). MAIN OUTCOME MEASURES: Cardiac size and LV diastolic function, peak oxygen uptake (Vo(2)), muscle strength, general and abdominal fatness, and insulin resistance. INTERVENTIONS: 6 months of exercise training versus usual care. RESULTS: When analysed by group at six months, cardiac size and LV diastolic function did not differ between exercisers (n = 51) and controls (n = 53), whereas exercisers had significantly higher peak Vo(2) (28 v 24 ml/kg/min) and strength (383 v 329 kg), and lower fatness (34% v 37%), diastolic BP (73 v 75 mm Hg) and insulin resistance (quantitative insulin sensitivity check index 0.35 v 0.34) versus controls (all p 90% (injection drug use). Uncontrolled HIV infection seems to accelerate the progression of HCV-induced liver fibrosis. Forty-eight weeks of combination therapy with pegylated interferon alpha (2a or 2b) plus ribavirin achieves a sustained viral response in coinfected individuals in up to 38% with HCV genotype 1 and up to 73% with genotypes 2 or 3. The safety profile of this treatment is similar to therapy in HCV-monoinfected patients with influenza-like symptoms, cytopenia and neuropsychiatric symptoms dominating. However, HIV/HCV-coinfected patients who also take zidovudine develop more profound anaemia than those on other HIV nucleoside analogue therapy. Didanosine and stavudine are associated with rare but serious mitochondrial toxicity, such as pancreatitis or lactic acidosis. It does not appear that the addition of ribavirin increases that risk. There is currently no evidence that in HIV/HCV coinfection one pegylated interferon product is superior to the other. Contrary to common perception, it is also unproven that HIV/HCV-coinfected patients respond less well to therapy with peginterferon alpha plus ribavirin than HCV monoinfected patients. Given the safety and efficacy of combination therapy with peginterferon plus ribavirin and the deleterious effects of chronic hepatitis C, all HIV/HCV-coinfected patients should be evaluated for therapy. PMID- 16308420 TI - The widespread impact of mammalian MicroRNAs on mRNA repression and evolution. AB - Thousands of mammalian messenger RNAs are under selective pressure to maintain 7 nucleotide sites matching microRNAs (miRNAs). We found that these conserved targets are often highly expressed at developmental stages before miRNA expression and that their levels tend to fall as the miRNA that targets them begins to accumulate. Nonconserved sites, which outnumber the conserved sites 10 to 1, also mediate repression. As a consequence, genes preferentially expressed at the same time and place as a miRNA have evolved to selectively avoid sites matching the miRNA. This phenomenon of selective avoidance extends to thousands of genes and enables spatial and temporal specificities of miRNAs to be revealed by finding tissues and developmental stages in which messages with corresponding sites are expressed at lower levels. PMID- 16308421 TI - The kinase LKB1 mediates glucose homeostasis in liver and therapeutic effects of metformin. AB - The Peutz-Jegher syndrome tumor-suppressor gene encodes a protein-threonine kinase, LKB1, which phosphorylates and activates AMPK [adenosine monophosphate (AMP)-activated protein kinase]. The deletion of LKB1 in the liver of adult mice resulted in a nearly complete loss of AMPK activity. Loss of LKB1 function resulted in hyperglycemia with increased gluconeogenic and lipogenic gene expression. In LKB1-deficient livers, TORC2, a transcriptional coactivator of CREB (cAMP response element-binding protein), was dephosphorylated and entered the nucleus, driving the expression of peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha), which in turn drives gluconeogenesis. Adenoviral small hairpin RNA (shRNA) for TORC2 reduced PGC-1alpha expression and normalized blood glucose levels in mice with deleted liver LKB1, indicating that TORC2 is a critical target of LKB1/AMPK signals in the regulation of gluconeogenesis. Finally, we show that metformin, one of the most widely prescribed type 2 diabetes therapeutics, requires LKB1 in the liver to lower blood glucose levels. PMID- 16308422 TI - Hf-W chronometry of lunar metals and the age and early differentiation of the Moon. AB - The use of hafnium-tungsten chronometry to date the Moon is hampered by cosmogenic tungsten-182 production mainly by neutron capture of tantalum-181 at the lunar surface. We report tungsten isotope data for lunar metals, which contain no 181Ta-derived cosmogenic 182W. The data reveal differences in indigenous 182W/184W of lunar mantle reservoirs, indicating crystallization of the lunar magma ocean 4.527 +/- 0.010 billion years ago. This age is consistent with the giant impact hypothesis and defines the completion of the major stage of Earth's accretion. PMID- 16308423 TI - Controversy of free radical hypothesis: reactive oxygen species--cause or consequence of tissue injury? AB - For a decade or two, the hypothesis of causality of various disorders by reactive oxygen species (ROS), due to their potentially harmful effect towards cellular constituents, is one of the most frequently cited in biomedical sciences. In fact, the ROS-mediated alterations of biomacromolecules are considered to be essential events in the etiopathogenesis of those diseases where involvement of ROS has been indicated. ROS easily react in vitro with most biological molecules, causing their degradation and destruction. This may implicitly suggest that, when excessively produced in vivo, ROS are deleterious to integral components of the cell and cause their dysfunctions. Some experimental data indicate that ROS mediated lipid peroxidation, protein oxidation and oxidative alterations to nucleic acids are crucial events of unfavorable actions of ROS. Yet the most convincing evidence, i.e. unambiguous inhibition of tissue injury by pretreatment with antioxidants, has not been provided. On the contrary, there are quite a few papers reporting failure in applying antioxidants to heal those pathologies where the causal role of ROS was supposed. Other papers reported serious complications arising from antioxidant therapy, which is quite in contradiction to its expected effect. On the other hand, an increasing number of recent findings have provided evidence of a key role of ROS in both intracellular signaling and intercellular communication, processes involved in maintaining homeostasis. Hence, some investigators consider excessive production of ROS to be rather a "smoke after the fire" than "a deleterious fire" itself, suggesting the occurrence of overproduced ROS as being the consequence of some primary damage. The present paper aims at summarizing some pros and cons of various opinions with an attempt to help better understand the involvement of ROS in tissue injury. PMID- 16308424 TI - pH-induced changes in activity and conformation of NADH oxidase from Thermus thermophilus. AB - Thermus thermophilus NADH oxidase (NOX) activity exhibits a bell-shaped pH dependency with the maximal rate at pH 5.2 and marked inhibition at lower pH. The first pH transition, from pH 7.2 to pH 5.2, results in more than a 2-fold activity increase with protonation of a group with pKa=6.1+/-0.1. The difference in fluorescence of the free and enzyme-bound flavin strongly indicates that the increase in enzyme activity in a pH-dependent manner is related to a protein cofactor interaction. Only one amino acid residue, His75, has an intrinsic pKa approximately 6.0 and is localized in proximity (<10 A) to N5-N10 of the isoalloxazine ring and, therefore, is able to participate in such an interaction. Solvent acidification leads to the second pH transition from pH 5.2 to 2.0 that results in complete inhibition of the enzyme with protonation of a group with an apparent pKa=4.0+/-0.1. Inactivation of NOX activity at low pH is not caused by large conformational changes in the quaternary structure as judged by intrinsic viscosity and sedimentation velocity experiments. NOX exists as a dimer even as an apoprotein at acidic conditions. There is a strong coupling between the fluorescence of the enzyme-bound flavin and the intrinsic tryptophans, as demonstrated by energy transfer between Trp47 and the isoalloxazine ring of flavin adenine dinucleotide (FAD). The pH-induced changes in intrinsic tryptophan and FAD fluorescence indicate that inhibition of the FAD-binding enzyme at low pH is related to dissociation of the flavin cofactor, due to protonation of its adenine moiety. PMID- 16308425 TI - Chronic exposure to constant light affects morphology and secretion of adrenal zona fasciculata cells in female rats. AB - The effect of chronic exposure to light of adult Wistar rats on growth and function of adrenal zona glomerulosa (ZG) and zona fasciculata (ZF) were examined. The females were exposed to continuous light of 600 lux for 95 days, starting on day 30 of age. The controls were kept under a 12:12 h light-dark cycle, at ambient temperature. The rats were sacrificed by decapitation and the left adrenal gland of each animal was dissected out and prepared for morphometric analyses. In animals exposed to chronic lighting, the absolute and relative volume of ZG were insignificantly increased by 5% (p>0.05) compared to controls. The volume of ZG cells and their nuclei were insignificantly changed by 1% (p>0.05) in comparison with corresponding controls. The absolute and relative volume of ZF were significantly increased (by 14 and 9%, respectively; p<0.05), as compared to controls. The volume of ZF cells and their nuclei were significantly increased (by 12 and 9%, respectively; p<0.05). Serum concentration of corticosterone was also significantly (p<0.05) increased by 13% in light exposed group in comparison with control rats. These findings suggest that continuous exposure of female rats to constant light increased growth and secretory activity of ZF cells. PMID- 16308426 TI - Effect of ajmaline on action potential and ionic currents in rat ventricular myocytes. AB - The effect of ajmaline on action potential (AP) and ionic current components has been investigated in right ventricular myocytes of rat at room temperature using the whole cell patch clamp technique. Ajmaline decreased the upstroke velocity ((dV/dt)max) of AP and the AP amplitude, increased the AP duration measured at 50 and 90% repolarization, and reversibly inhibited most components of membrane ionic current in a concentration-dependent manner. The following values of IC50 and of the Hill coefficient (nH) resulted from approximation of the measured data by the Hill formula: for fast sodium current (INa) IC50=27.8+/-1.14 micromol/l and nH=1.27+/-0.25 at holding potential -75 mV, IC50=47.2+/-1.16 micromol/l and nH=1.16+/-0.21 at holding potential -120 mV; for L-type calcium current (ICa-L) IC50=70.8+/-0.09 micromol/l and n(H)=0.99+/-0.09; for transient outward potassium current (Ito) IC50=25.9+/-2.91 micromol/l and nH=1.07+/-0.15; for ATP-sensitive potassium current (IK(ATP)) IC50=13.3+/-1.1 micromol/l and nH=1.16+/-0.15. The current measured at the end of 300 ms depolarizing impulse was composed of an ajmaline-insensitive component and a component inhibited with IC50=61.0+/-1.1 micromol/l and nH=0.91+/-0.08. At hyperpolarizing voltages, ajmaline at high concentration of 300 micromol/l reduced the inward moiety of time-independent potassium current (IK1) by 36%. The results indicate that the inhibition of INa causes both the decreased rate of rise of depolarizing phase and the lowered amplitude of AP. The inhibition of Ito is responsible for the ajmaline-induced AP prolongation. PMID- 16308427 TI - In vitro study of astrocytic tumour metabolism by proton magnetic resonance spectroscopy. AB - In vivo magnetic resonance spectroscopy (MRS) studies of glial brain tumours reported that higher grade of astrocytoma is associated with increased level of choline-containing compounds (Cho) and decreased levels of N-acetylaspartate (NAA) and creatine and phosphocreatine (Cr). In this work, we studied the metabolism of glioma tumours by in vitro proton magnetic resonance spectroscopy (1H-MRS). 1H-MR spectra were recorded in vitro from perchloric acid extracts of astrocytoma (WHO II) and glioblastoma multiforme (WHO IV) samples. We observed differences between astrocytoma and glioblastoma multiforme in the levels of Cho, alanine, lactate, NAA, and glutamate/glutamine. In astrocytoma samples, we found higher MR signal of NAA and lower signal of Cho and alanine. MR spectra of glioblastoma samples reported significantly higher levels of lactate and glutamate/glutamine. In contrast, levels of Cr were the same in both tumour types. We also determined NAA/Cr and Cho/Cr ratios in the tumour samples. The NAA/Cr ratio was higher in astrocytomas than in glioblastomas multiforme. Conversely, the Cho/Cr ratio was higher in glioblastoma multiforme. The results indicate that MRS is a promising method for distinguishing pathologies in human brain and for pre-surgical grading of brain tumours. PMID- 16308428 TI - Computer-aided formation of the whole-cell patch-clamp recording configuration. AB - The conventional patch-clamp technique requires well-trained experimenter. Few commercial automated patch-clamp systems, designed for drug development, are better suited for large-scale research then for standard electrophysiological experiments. Here we describe a state machine for automated recognition of recording states of the patch-clamp experiment. The principle of the state machine is based on evaluation of the charge carried by membrane current during specific time segments in responses to square wave voltage stimulation. The state machine may serve for generating various sound alerts, signals for automated control of other devices, assistance in micromanipulation, internal pipette pressure control, and holding potential adjustments. Algorithm of the state machine, designed to cover wide variety of cell types, was successfully tested on rat ventricular myocytes. PMID- 16308429 TI - Identification of all alpha1-adrenoceptor subtypes in rat lung. AB - The function of lung tissue is regulated via a release of neurotransmitters from autonomic nerves. The neurotransmitters of sympathetic nervous system, adrenaline and noradrenaline, activate both alpha-adrenoceptors and beta-adrenoceptors. Although the function and expression of beta-adrenoceptors can be considered major, some doubts exist about the function and expression of alpha1-adrenoceptor subtypes in the lung tissue. Therefore a set of competition binding experiments was employed in order to discriminate between the alpha1-adrenoceptor binding site subtypes in the rat lung. We identified three subpopulations of alpha1 adrenoceptor binding sites in the rat lung (alpha1A, alpha1B and alpha1D). PMID- 16308430 TI - Ischemia-reperfusion injury--antiarrhythmic effect of melatonin associated with reduced recovering of contractility. AB - The effect of melatonin on reperfusion arrhythmias and postischemic contractile dysfunction was studied in the isolated rat heart. 25 min global ischemia was induced and followed by 30 min of reperfusion. Melatonin (10 micromol/l) was present in the perfusion solution during the whole experiment. Experiment revealed protective effect of melatonin on reperfusion-induced arrhythmias- arrhythmia score was significantly lower as well as the total time of arrhythmias duration was significantly shorter in melatonin group than in controls. On the other hand, post-ischemic recovering of contractility was significantly reduced in melatonin group. PMID- 16308431 TI - Several aspects of formation of auto-transplant and re-transplant vascular pedicle. AB - The purpose of the work was the simplification of technical side of autotransplantation and replantation by pre-preparation of vascular pedicle. 173 patients were observed and treated. Microsurgical autotransplantation of tissues was done in 29 cases, fingers autotransplantation in 44 cases (35 patients), fingers replantation in 66 cases (38 patients), replantation of thick parts of extremities in 34 cases (3 patients). Thoracodorsal flap on the neuro-vascular flap was transplntated in 16 cases, groin flap in 3 cases, deltoid flap -- in 3 cases, scapular flap -- in 1 case, saphenal -- 1 case, gracilis muscle in 3 cases, iliac crest flap in 2 cases, rib segment on vascular pedicle -- in 8 cases. Rib segment was transplanted as an isolated and with pre-prepared thoracodorsal bone-soft tissue complex also. II toe-finger and II-III toe-fingers block were transplanted for the abnormalities of the hand. Good and satisfactory functional and aesthetic results were obtained in all cases of autotransplantation and replantation. Original method of anastomosing of vessels was used in vein transplantation. We could simplify and increased the number of good results of operations by pre-preparing vascular pedicle. The results of treatment were improved by optimal election of donor site and preliminary forming of vascular pedicle of flaps and replants. PMID- 16308432 TI - [Surgical treatment of the scalenus anticus syndrome]. AB - The purpose of this study was to assess the results of decompression of the thoracic outlet due to scalenus anticus syndrome and thus to give recommendation for a safe and effective surgical treatment. Over a 20-years period 100 patients (125 operations) underwent thoracic outlet decompression. There were 77 females and 23 male, with a median age 29 years (13-45), 4 pediatric patients were under 16 years. Of the 125 affected limbs, the indications for surgery were combination of both neurological and vascular symptoms in 55 patients, neurological symptoms in 21, and 24 patients had vascular symptoms alone. 54 patients had a transient ischemic attacks (TIA) due to the disturbance of the blood flow in the vertebral arteries, and 4 cardialgia due to the reduced blood flow in the thoracic internal arteries. From the results of this study we concluded that supraclavicular scalenectomy is a safe and effective procedure for most patients with scalenus anticus syndrome. PMID- 16308433 TI - [Correlational studies of urine interleukin 6, interleukin 8 and nitric oxide in the patients with pyelonephritis and hydronephrosis]. AB - The urine concentration of IL-6, IL-8 and nitric oxide (NO) were determined in the patients with pyelonephritis and hydronephrosis. Correlations between urine levels of IL-6 and IL-8 and amount of nitric oxide in the hydronephrosis patients were not found. However, in the patients with both acute and chronic pyelonephritis the coefficient of correlation was high, r((IL-6/NO)) = 0,94 and r((IL-8/NO)) = 0,86 for acute and r((IL-6/NO)) = 0,72 and r((IL-8/NO)) = 0,40 for chronic forms, respectively. These data suggest that secretion of NO during hydronephrosis has a compensatory character and acted on renal microvascular tone, whereas during pyelonephritis NO produced inflammatory mediators by recruiting leukocytes and has pathogenic character. PMID- 16308434 TI - [Comparative assessment of the quality of life of reproductive age patients with endometriosis depending on the method of treatment]. AB - The purpose of the study was to investigate the influence of endometriosis as well as certain widely-spread methods of its conservative hormonal and surgical treatment on the quality of life of the patients in reproductive age. 546 twenty to forty years old women have been examined: 371 patients with the first time diagnosed endometriosis (I group), 64 -- after conservative hormonal therapy (II group), 111 -- after radical surgery (III group) and 30 healthy women (control group). All women operated for endometriosis were assessed to determine the quality of life by the following scores: "unsatisfactory" -- 0 point, "satisfactory" -- 1 point, excellent -- 2 points. The average index of quality of life among all subjects was equal to 0,5+/-0,62 points (from 1,0+/-0,69 in the group of patients after one-sided ovariectomy or adnexectomy to 0,3+/-0,45 -- in patients after double adnexectomy with hysterectomy) that was 7,8 times (p<0,001) and 2,6 times less than the given indices in the control group and in the untreated patients correspondingly. So, endometriosis in reproductive age significantly deteriorates the quality of life of the patients in comparison with healthy women. This quality can't be fully restored by any of the analyzed methods of conservative hormonal treatment, radical surgery leads to its further significant deterioration requiring thus to continue searching for the most optimal approaches to the treatment and rehabilitation of patients with endometriosis in the reproductive age. PMID- 16308435 TI - Hospital pneumonia following cardiac surgery in children. AB - Any massive surgical intervention, especially cardiovascular surgery creates high risk for the development of hospital infections. Hospital pneumonia is the major cause of morbidity and mortality. The aim of our study was to determine the frequency and etiology of hospital pneumonia following cardiac surgery in children and to find out whether the emergency operations and reoperations increase the frequency of hospital pneumonia. We studied 355 patients in Jo Ann Pediatric Cardiac Surgery Clinic, who were operated in the period from January 2002 till December 2004. The age of the patients varied from 1 day to 18 years. Out of 355 operations 290 were primary ones, 65 reoperations. In 320 cases the operations were planed, 35 cases-emergency ones. Out of 355 patients, hospital infection occurred in 42 (11,9%) cases. The most common infectious complication was pneumonia that made up 18 cases (5,1%). In 77,8% of cases pneumonia was associated with mechanical ventilation. Hospital pneumonia in 60% were caused by gram-negative, in 20% -- by gram-positive bacteria and in 20% -- by fungi. The most frequent gram-negative bacterium were nonfermenter gram-negative rods, among these pathogens Stenotrophomonas maltophilia was the most common and the most frequent gram-positive bacterium was Staphylococcus aureus. All three cases of fungal pneumonia were caused by Aspergillus spp. The rate of hospital pneumonia was significantly higher in the cases of emergency operations as well as reoperations. PMID- 16308436 TI - [Mortality case analyses of obstetric-gynecologic sepsis]. AB - The aim of the study was the clinical and pathology description of patients who died due to post-delivery or postabortion sepsis. We performed the retrospective analysis based on the postmortem examination of patients who died after post delivery or postabortion sepsis at the Anti-Sepsis Center during 1987-2000. For thirteen years at gynecological department of the Anti-Sepsis Center out of 524 patients hospitalized with the diagnosis of postpartum or postabortion sepsis 45 died (8.6%). During all those years lethality fluctuated between 2.2%-15.5%. The existence of extragenital diseases in pre-delivery period was afterwards the risk factor for the development of polyorganic insufficiency with sepsis. These irreversible changes was the cause lethal outcome. The main factors were delayed hospitalization, severity of complications and septic shock. PMID- 16308437 TI - [Role of psychological factors in the developments of heart disease]. AB - The main purpose of the study was the evaluation of the role of various types of the fixed predisposition in the patients with ischemic heart disease (IHD) and arterial hypertension (AH) in different groups of patients to find statistically significant associations between the types of fixed predisposition and risk factors of IHD. Three groups of patients were investigated: 72 patients with AH, 175 patients with IHD ( 74 with angina pectoris and 101 with angina pectoris and myocardial infarction) and 164 patients with IHD and AH. Reliable relationship were found between crude-dynamic type of the fixed predisposition and IHD and AH. No significant associations were revealed between types of the fixed predisposition and well-known risk factors of IHD and AH. On the basis of our investigations it may be concluded that the specific types of the fixed predisposition represent independent risk factors of IHD and AH. Determination of such risk-factors has great importance for the risk assessment of the heart diseases. PMID- 16308438 TI - [A type behavior as a risk factor in siblings with diabetes mellitus]. AB - The objective of the study was to investigate the prevalence of the "a" type behavior among close relatives (including siblings) of the subjects with diabetes mellitus. For this purpose there were examined 50 practically healthy siblings aged between 20 and 40, among them 27 (54%) females and 23 (46%) males. The method of the structural interview was applied. In addition to this, they were subjected to the tests of glucose tolerance. The considerable number of the siblings (46%) showed the "a" type behavior of various kinds, and 44% -- the disruption of glucose tolerance, which produces the preliminary conditions for the diabetes manifestations and gives us the grounds for applying further preventive activities. PMID- 16308439 TI - [QT interval variability in adjacent areas on standard ecg during ectopic activity in patients with acute coronary syndrome]. AB - Clinical and experimental evidence closely link malignant ventricular arrhythmia with increased dispersion of refractoriness and thus electrical heterogeneity in different kinds of heart diseases. Most increased dispersion of refractoriness is due to inhomogenous action potential duration in adjacent areas, decreased local conduction velocity or both. The article focuses on ectopic ventricular activity and on QT interval variability in adjacent segments on standard ECG in patients with acute coronary syndrome. The adjacent QT dispersion was defined as a difference between two adjacent leads. Seventy nine patients aged 54+/-6,5 in 6 hour interval after the attack with elevation of ST segments on ECG were studied. Fifteen patients aged 48+/-7 without elevation of ST segment on ECG served as control subjects. Patients were allocated to two groups: one included patients with 1-2 ectopic degree; group two included patients with 3-5 ectopic degree. In patients with acute coronary syndrome QT max and QTd was significantly higher than in control subjects (p<0,05). The increase of ventricular ectopic degree is linked with the increased adjacent QT dispersion. PMID- 16308440 TI - [Activity of lipid peroxidation processes and the condition of antioxidative defense system in children with rheumatic fever]. AB - Aim of study consists of establishing of some clinical-biological correlates for rheumatic fever (RF) in children, namely correlations between clinical status and lipoperoxidation products -- malonidialdehide (MDA) and antioxidative enzymes in the blood. In the neutralization process of superoxside anions ceruloplazmin (CP), catalase (CAT) and transferin (TF) are key antioxidant enzymes (AOE) of antioxidative defense system (AOD). We studied 38 patients 3-15 years of age with different variants of RF. We measured the levels of MDA, CAT, CP, TF in the plasma in patients with acute rheumatic fever (ARF) and chronic rheumatic heart disease (CRHD). In all of our cases AOE (CAT, CP, TF) were high at time of diagnosis, concomitant with increased MDA and inflammatory tests. Our study revealed intensified activity of AOD enzymes in children with RF. PMID- 16308441 TI - Health-related interests of Georgian adolescents. AB - The aim of the study was to find out what adolescents in Georgia would like to know about health. Pupils of 6-10 grades (11-17 years old) (n=524) of the focus schools (secondary schools where FRESH program is being implemented) were investigated. A method of participatory research (so called social anthropologic approach) was applied. The study was anonymous. It yielded 3756 questions in total. The questions dedicated to health-related issues made up 36.8% for group I (11-13y.o.), 45,75% for group II (14-17y.o.), 40.5% -- total. The interest in different somatic and psychological problems and diseases was the same in both age groups (14,3%). Interest in sexual items was higher in later adolescents (group I -- 10.58+/-0.66%, group II -- 14.97+/-0.90%, p<0.05). Girls have shown more interest in diseases and common medical problems (M -- 27.3+/-3.2%, F - 45.2+/-2.6%; p<0.05). Boys were more interested in sexual behavior (M -- 40.2+/ 3.6%, F -- 25.2+/-2.3%; p<0.05). There was shown a purposefullness of applied research methodology. PMID- 16308442 TI - [IFN-gamma in non atopic children with recurrent obstructive bronchitis]. AB - Recurrent respiratory infection cause an imbalance of Th1/Th2 immune response with decreased level of IFN-gamma. Result of several studies have provided evidence linking Mycoplasma infection with recurrent wheezing in atopic children. The aim of the present study was to investigate the influence of Mycoplasma infection on IFN-gamma level in non-atopic children with recurrent obstructive bronchitis. Serum IFN-gamma was measured in two groups: the study group included 30 non-atopic children 1-4 years of age with recurrent obstructive bronchitis caused by Mycoplasma pneumoniae, the control group included 15 children with the same disease with negative Mycoplasma pneumoniae result. There were no differences in age, sex and family history of atopy. This result shows reduced Th 1 immune response that causes reduced antiviral activity and high morbidity in non-atopic wheezy children. PMID- 16308443 TI - [Spirometric values of asthmatic children treated with Plaferon-LB]. AB - A long-term (3 week-lasting), double-blind, placebo-controlled study was carried out in order to determine the efficacy of inhaled Plaferon LB (a 6 Kd protein fraction-peptide, obtained by purification of human placental amniotic membrane) in 43 asthmatic children. Study group of patients were receiving daily inhalations (using special nebulizer) of a standard dose of plaferon (5 mg/ml - 3 ml), while in placebo group an inert solution was used. At the end of each trial week the spirometric data of patients were obtained. According to the results of the study, plaferon-inhalation therapy was associated with statistically significant improvement of spirometric values. It may be concluded long-term plaferon treatment provides an effective and safe alternative to traditional bronchodilator drugs in the management of childhood asthma. PMID- 16308444 TI - Wobenzym in treatment of recurrent obstructive bronchitis in children. AB - A total of 27 patients with recurrent obstructive bronchitis (at least 3 episodes of obstructive bronchitis per year) of 5-15 years of age were studied. Serum IgE level (by ELISA method) and pulmonary function indices were determined together with the full clinical examination. The systemic enzyme preparation Wobenzym was administered with a dosage of 1 tablet for 6 kg weight. Wobenzym was administered for 3 months. Effectiveness of the treatment was assessed by special questionnaire, in which existence of the main symptoms were recorded. Analysis of the data, obtained after treatment, demonstrated decrease of the Daily Symptom Score and an increase of Symptom Free Days, as well as an improvement in spirometric indices (FVC, FEV, PEF). According to these data, it was concluded, that systemic enzyme preparation Wobenzym should be used as a supporting measure in combination treatment of recurrent obstructive bronchitis. PMID- 16308445 TI - [Comparative evaluation of microbial colonization of newborns according to the working conditions of maternity houses]. AB - This study was aimed on the investigation of microbial colonization of 57 newborns at maternity home, applying the working program "BFH" ("Friendly attitude to child" -- a system of joint requirement of mother and child with respect to introducing the method of early delivery to breast-feed), and microbial colonization in 45 newborns and their mothers at ordinary maternity home, where they were being held apart from each other. The final results of the investigation showed up the following: the microbial colonization of newborns at the departments, applying the "BFH" program as working system, presented to be more than 2 times less intensive, compared to the newborns, held apart from their mothers. The gram-positive microflora (epidermal staphylococci, in 30% of cases they demonstrate the hemolytic features) and the culture of C. albicans were manifested to be prevalent in the microbial structure of newborns. The outcomes of the investigation give the base for recommendations for full engagement of maternity homes in the working system based on the principle of "mother-child" and family-oriented obstetrics. PMID- 16308446 TI - Morphological pre-conditions of diabetes mellitus' development under chronic lipid-loading during aging. AB - Chronic lipid loading in pancreatic beta-cells of young and old animals causes the intensification of secretion the morphologic equivalents of which are revealed by elevation of number and sizes of mitochondria, granular endoplasmatic reticulum and Golgi apparatus of beta-cells with the quantitative increase of intraorganelle ultrastructures and decrease of volume share and number of secretor granules, which are more prominent in old animals. After definitive period following the chronic lipid loading the secretor processes in young age are normalized (organelles turn back to their initial volume state and consist of the same initial number of intraultrastructures), whereas in old age -- are markedly lower compared with the norm (size and number of organelles and their inner structures are decreased; volume part of secretory granules is increased); in some part of beta-cells complete block of secretion takes place (in the part of beta-cells the irreversible changes are developed; stagnation of secrete is prominent). Therefore any metabolic disease or syndrome running on the background of chronic lipidemia, seems to be a risk-factor for development of Diabetes Mellitus in organisms of old age. PMID- 16308447 TI - Ultrastructural organization of epiphysis in rats under the action of electromagnetic fields and during mammary carcinogenesi. AB - Experiments conducted by the authors, as well as clinical studies, show that in addition to hormonal mis-balance, development of mammary gland cancer is significantly influenced by the action of low frequency electromagnetic fields on epiphysis. By reducing the production of melatonin it increases the risk of development of mammary gland tumors. The review of scientific literature indicates that pinealocytes are the main morphological substrate responsible for functional activity of pineal cells. When estimating large specific weight of lipids, many researchers point to the dependence of their presence in epiphysis on the level of melatonin. It is thought that hormones of pineal cells are deposited in the form of lipid drops. The most characteristic feature in the structure of pinealocytes is the presence of large number of various size complexes in their bodies and appendices. According to many authors this could be the form of depositing the secretory products of pinealocytes, such as melatonin, biological amines, etc. Ultrastructure characteristic of pinealocytes described in the review provides deeper understanding of the fine structure morpho physiology of epiphysis, enable to point out the peculiarities of its functionality under the influence of electromagnetic fields and discover the organizational structure of pineal body at a time of the mammary gland tumor development. PMID- 16308448 TI - [Homocysteine as risk marker of cardiovascular disease]. AB - Basic research has demonstrated that homocysteine enhances both arteriosclerosis and thrombosis - the principal cause of CVD. Although studies demonstrate that patients with elevated homocysteine levels have small to moderate increased risk of CVD it remains unclear whether lowering plasma homocysteine levels will decrease risks of CVD. The study was aimed to detect reliability of homocysteine lowering on risk of CVD. We studied 256 patients (56 stenocardia; 96 myocardial infarction; 104 post infarction period). Besides we investigated the influence of blood homocysteine concentration on coronary vessels of patients suffering ischemic heart disease, acute heart disease and those in post infarct period. The majority of coronary vessels demonstrated occlusion and stenosis. The investigation showed the connection between body reactivity and homocysteine level. The higher body reactivity was associated with the higher homocysteine concentration in the blood. It stimulated the development of the mild form of ischemic heart disease -- stenocardia. The increase of homocysteine concentration up to the normal top level points to the danger of development of acute ischemic myocardial disease rather than stenocardia. The investigation showed that the homocysteine blood levels are among the reliable diagnostic markers of CVD. The results of the investigation would permit rational clinical decision making for individual patients and policy decisions for the health of the general population. PMID- 16308449 TI - [Aminoguanidine decreases the brain damage in rats neonatally exposed to hypoxic ischemic influences]. AB - Seven days old rats (n=30) were divided into five groups (6 animals in each). Three groups underwent hypoxic-ischemic event (occlusion of right common carotid artery lasting 2,5 hours and breathing with 8% oxygen and 92% of nitrogen mixture). The rats of the fifth group, after completion of hypoxic-ischemic impact and following a daily reoxygenation (for three days) were injected (i/p) with selective inhibitor of nitric oxide inducible synthase, in a dose of 300 mg/kg. The rats of the fourth group were injected with 150 mg/kg of the selective inhibitor of nitric oxide synthase -- aminoguanidine (150 mg/kg); the animals of the third group were injected with saline (50 ml). The second group contained the shame-operated animals and the first group -- intact (control) animals. An intensity of local cerebral blood flow in fronto-parietal area of neocortex was measured by means of hydrogen clearance technique. In the right hemisphere (ipsilateral to occluded common carotid artery) of the animals of the third group significant decrease, and in contralateral hemisphere (left) temporal increase of CBF did occur. Administration of aminoguanidine prevents high degree of brain damage observed in the control group of animals. PMID- 16308450 TI - [The action of dextrorphan and sigma ligands on the spontaneous secretion of interleukins by jurkat cell line]. AB - Sigma receptor was demonstrated to have at least two subtypes, mediating pharmacological effects of various preparations including psychoactive, neuroleptic, cardioprotector, anti-inflammatory, immunosuppressive compounds and several steroid hormones. The stimulation of sigma receptor induces transient increase of intracellular calcium and amplifies signals from different stimuli. Pentazocine, SKF 10 047, dextrorphan, and other sigma ligands including phencyclidine and haloperidol were investigated for their potential immunoregulatory properties. We have found, that pentazocine, SKF 10 047, dextrorphan reduce spontaneous secretion of IL-8, IL-6 and IL-10 and selectively changes synthesis of IL-4 by Jurkat human T lymphocyte cells lines. Dextrorphan significantly enhanced, pentazocine, haloperidol and phencyclidine had no effect, while SKF 10 047 suppressed production of IL-4. Spontaneous secretion of IL-4 and IL-8 correlates with synthesis of nitric oxide, suggesting that NO and transitory S-nitrosylation of up-stream proteins participate in the sigma ligand dependent expression of IL-4 and IL-8 genes. PMID- 16308451 TI - [The immunohistochemical characteristics of cervical glandular intraepithelial neoplasias]. AB - The aim of our study was to identify immunohistochemical diagnostic criteria for cervical glandular intraepithelial neoplasias (CGIN). We examined 136 women with cytological diagnosis of atypical endocervical cells. These patients were divided into three groups based on a grade of the lesion: 35 patients with CGIN1 (group I), 72 patients with CGIN2 (group II), 28 patients with CGIN3 (group III). Endocervical curettages were examined by hematoxylin-eosin and immunohistochemistry using monoclonal antibodies against Ki-67, EpAg, MNF116, CEA, EMA. We used histological algorithm created by us. The comparative analysis of immunohistochemical results showed that expression of Ki-67 is seen in CGIN 2 and significantly increased in CGIN3 (p<0,05) which indicates increased proliferative activity of glandular cells in relation to increased grade of lesion. The differences in the expression of MNF116 and EMA are not statistically significant (p>0,05) which indicates that the expression of these epithelium specific markers does not change according to the grade of atypia and carcinogenesis (they can be used for determination of tumor phenotype). The expression of CEA and EpAg is strongly increased in CGIN2 and CGIN3 (p<0,05) indicating their potential role in carcinogenesis. The results suggest that evaluation of a grade of cervical glandular intraepithelial neoplasia should be based on histological and immunohistochemical studies. The morphometric algorithm should include the following criteria: type of lining epithelium (cubical, columnar), nuclear cytoplasmic index (<1, >1, =1), stratification, hyper- and hypochromasia, size and amount of nucleoli, and stromal-parenchymal ratio. The immunohistochemical study should include the expression of proliferation marker (Ki-67), carcinoembryonic antigen (CEA)and Epithelial Antigen (EpAg). We recommend the classification of CGIN into two types: low grade cervical glandular intraepithelial neoplasia (CGIN 1) and high grade cervical glandular intraepithelial neoplasia including CGIN 2 and CGIN 3. PMID- 16308452 TI - New method for treatment of high hepaticocholedoch strictures (experimental research). AB - The purpose of the investigation was to achieve hepaticocholedoch recovery with preserved sphincter apparatus. The experiments were carried out on 25 mongrel dogs of both sexes with body weight 12-25 kg. The animals were divided into 3 groups. The hepatocholedoch strictures were induced in the animals of group I (n=10) and II (n=10). On the 4th day following modeling the animals of group II were subjected to relaparatomy, stricture removal and hepatocholedoch distraction by means of distraction device developed by prof. Kakabadze (patent request submitted for approval). On the 18th day after the surgical procedure the animals have undergone the surgery, the distraction device was removed and the anatomical integrity of biliary tract was restored by bilio-biliary anastomosis. Performed investigations have shown that all of the animals of group I have died on the several time points after the surgery. The cause of death was cholestasis. As to animals of group II, 2 dogs have died on 24 and 35 days after the surgery because of technical shortages of the bilio-biliary anastomosis. The rest 8 experimental animals undergo surgery and postoperative period normally and they were under observation in vivarium standard conditions correspondingly. The i.v. cholangiography, performed in survived animals during postoperative period, showed compete contrast liquid flow into the duodenum. The histological test performed during distraction showed mucosal layer sclerosis. Distraction and afford made evident influence on the muscular fibers through the duct axis. Their elongation and attenuation was mentioned after 3 days from distraction. The fibroblasts local accumulation was seen. The hepaticocholedoch mucosal complete recovery was seen after apparatus was taken out and the bilio-biliary anastomosis was done. Thus, new method of hepaticocholedoch strictures surgical correction elaborated by us allows us to restore anatomical integrity of biliary tract with preserved Oddi sphincter. PMID- 16308453 TI - [Sensory and psychoemotional productivity in the type I diabetes mellitus]. AB - Depressive psychosis with anxiety or without it is the most spread disorder among the patients with diabetes mellitus. According to our investigations the depression was found in 36.6% of cases of patients with the I type diabetes mellitus which affects negatively their mental working ability. Based on the mentioned above we aimed to study a sensory and psychoemotional productivity of patients with the type I diabetes mellitus for their further purposeful treatment. Thirty patients with the type I diabetes mellitus were observed and divided into two groups -- 19 -- without and 11 with a depression. Sensor and motor reaction was studied by means of Landolt's rings in computerized mode. A deterioration of the patients' mental working ability while resolving the simple sensor and motor task is expressed as the increased quantity of missed reactions on the light stimulus and chaotic character of the reaction time histograms that is stipulated by the diminished ability to concentrate attention be carrying out a monotonous work. Psychoemotional test has proved our conclusion drawn by means of sensor-motor task, but the analysis of the reaction time histograms allows suggesting that given contingent of patients fulfill the task of the various conditions of resolving easier then the monotonous one. PMID- 16308454 TI - [Organisational aspects and existing problems in prevention and early diagnostics of lung cancer]. AB - Thorough study of advanced stages of lung cancer has been held. Delayed detection of most of the malignant tumours (IV stages) points to a number of problems existing in Georgia in terms of primary prevention and early detection of oncologic diseases. Hence, elaboration of strategy for prevention and early detection of oncologic diseases is of great importance for our country. Activities aimed at early detection of lung cancer under the conditions of low level resources primarily imply education of population and professionals, as well as implementation of screening procedures. Cheap and effective methods should be applied for early diagnosis of one or two most widespread malignant tumours. PMID- 16308455 TI - [Complex study of the problem on accessibility to high quality medical care in the population of Georgia]. AB - The problem of accessibility to quality medical care, involving quality improvement in primary and hospital care, professional development of health professionals, protection of consumer rights, confidentiality of the information on the health of the patient, etc. still remains a serious problem for the health system of Georgia. Study was based on the long-term observation of keeping the standards of quality of care in frames of providing high qualification and skills of physicians, working in primary and hospital care and on the detailed analysis of currently available material. Based on the carried out study authors believe that it is necessary to develop in the nearest future formal manuals on medical care based on the Evidence-Based Medicine. As an obligatory condition the authors recommend to establish a system, in which the physicians should have easy access to the data of clinical epidemiology, up-to-date international information on new technologies, and information on comparative analysis of their effectiveness in our country. The integral part of such formal manual should be the identification of the facility for providing care in case of given disease/condition in accordance to the licensed types of care at different level facilities, i.e. facilities, having appropriate equipment and staff. This will solve the problem of fulfillment the state guarantees on providing equity in health care despite to the place of residence. Evaluation, assurance and control of the quality of care on all managerial levels (federal, regional, rayon, facility, unit) should be provided by unified principle of the priority of the criteria of the quality of outcomes (intermediate and final). PMID- 16308456 TI - Quantitative analysis of biochemical characteristics of bone-patellar tendon-bone allografts. AB - Allografts were used in the 80s and 90s to replace ruptured anterior cruciate ligaments (ACL) in the reduction of donor site morbidity. It was evident that in comparison with the autologous cruciate ligament grafts (ligamentum patellae) using Allografts (Tutoplast), a noticeably higher rupture rate appeared. The results from the work submitted serve as a detailed, highly structured description of human bone-patellar, tendon-bone (BPTB) and allografts (Tutoplast), which were used to replace ruptured anterior cruciate ligaments. The biochemical parameters: hydroxyproline or, as the case may be, total collagen content and non-reducible crosslinks were quantified. The examined material consisted of allografts, which had been taken from human ligamentum patellae and implanted five years ago as cruciate ligament reconstructions. The total collagen content in the analysis material amounted to 52.74%. The pyridinoline concentrations indicated in the test material amounted to 0.2135 mol PYR/mol collagen. The tissues examined contained 0.009245 mol DPD/mol collagen. The results show that in comparison with the values given for these tissues in the literature, there was a clear reduction in the total collagen content and the concentration of non-reducible correlations. On the basis of the analysis it may be supposed that the changed biochemical parameters are a cause of the increased rupture rate in allografts. PMID- 16308457 TI - In vitro regulation of single collagen fibril length by buffer compositions and temperature. AB - An understanding of collagen ultrastructure is very important for designing biopolymers mimicking collagen functions in tissue engineering, or for diagnosing abnormal collagen structure in clinical study. The present study examined formation of a large population of type I collagen single fibrils under different buffer compositions and temperatures. Fibril structures were investigated by dark field microscopy and atomic force microscopy (AFM). In the phosphate buffered saline (PBS) buffer, the average lengths of single fibrils were 4.8+/-2.2, 5.0+/ 1.9 and 9.2+/-5.0 microm for 37 degrees C, 33 degrees C and 29 degrees C, respectively. The differences were significant (P < 0.05) between 37 degrees C and 29 degrees C and between 33 degrees C and 29 degrees C. In the sodium phosphate (SP) buffer, the average lengths of single fibrils were 10.6+/-5.4, 11.1+/-4.5 and 19.6+/-11.7 microm for 37 degrees C, 33 degrees C and 29 degrees C, respectively. Similarly, the differences were significant (P < 0.05) between 37 degrees C and 29 degrees C and between 33 degrees C and 29 degrees C. While at the same temperature, the average lengths of single fibrils differed significantly (P < 0.05) between PBS and SP buffers. Single fibrils formed in SP buffer were found to have greater average length than those formed in PBS buffer. PMID- 16308458 TI - Experimental evaluation of the magnetic properties of commercially available magnetic microspheres. AB - The magnetic properties of 5 commercially available magnetic microsphere samples are tested and compared with those stated by their manufacturers. A suspension of magnetic, iron oxide nanoparticles is studied for comparison. Two of the microsphere samples have magnetic properties which do not support the manufacturer's claims of superparamagnetism. The remaining 3 microsphere samples as well as the nanoparticle suspension are superparamagnetic or ferromagnetic as claimed by the manufacturers. Field cooled and zero field cooled magnetisations indicate that the non-superparamagnetic microsphere samples contain blocked magnetic particles at room temperature. This observation is supported by the open hysteresis loops of the room temperature, field dependent magnetisation measurement. There is a significant paramagnetic component in the superparamagnetic microspheres. This is also present to a lesser extent in a nanoparticle suspension. PMID- 16308459 TI - Fixation stiffness of Dynafix unilateral external fixator in neutral and non neutral configurations. AB - A primary function of external fixator is to stabilize the fracture site after fracture reduction. Conventional fracture reduction method would result in fixator configurations deviated from its neutral configuration. How the non neutral configurations would affect the biomechanical performance of unilateral external fixators is still not well-documented. We developed a finite element model to predict the fixation stiffness of the Dynafix unilateral external fixator at arbitrary configurations under compression, torsion, three-point, and four-point bending. Experimental testing was done to validate the model using six Dynafix unilateral external fixators in neutral and particular non-neutral configurations. Effects of loading directions on bending stiffness were also studied. It appeared that the model succeeded in revealing the relative stiffness of the neutral and non-neutral configuration in all the loading conditions. Our results also demonstrated that bending stiffness could vary substantially for different loading directions and the principle loading directions could be very different for different fixator configurations. Therefore, a more logical way to compare the bending stiffness is to identify the principle loading directions of each fixator configuration and used their maximum and minimum bending stiffness as comparison criteria. Given that fixator configurations could substantially change the stiffness properties of the bone-fixator system, computer simulation with finite element modeling of this kind will provide useful clinical information on the rigidity of certain configurations in stabilizing the fracture site for bone healing. PMID- 16308460 TI - Short-term human chondrocyte culturing on oriented collagen coated gelatine scaffolds for cartilage replacement. AB - The biological, biochemical, mechanical, and structural properties of artificial scaffolds for tissue engineering are known to be of great importance. Therefore, in this study a hydrogel derived scaffold with biomechanical and structural properties similar to native articular cartilage was synthesized. The gelatine based hydrogel was processed by freeze-structuring, structuring by electrochemical water-decomposition, freeze-drying and chemical fixation resulting in a defined scaffold-structure. By electron microscopy a perpendicular pore-channel structure was verified with channel diameters between 30 microm and 70 microm. Mechanical testing showed mechanical properties similar to native cartilage. Human chondrocytes from biopsy samples were cultivated on these hydrogel scaffolds for three days. Two different cell densities (1.2 x 10(6) cells/cm3 and 12.0 x 10(6) cells/cm3) were used for cultivation. Histology of the cell seeded artificial scaffolds demonstrated vital cells that are widely distributed within the scaffold and mimic a columnar arrangement. PMID- 16308461 TI - Determination of the intracellular Ca2+ concentration in the N1E-115 neuronal cell line in perspective of its use for peripheric nerve regeneration. AB - Entubulation repair of peripheral nerve injuries has a lengthy history. Several experimental and clinical studies have explored the effectiveness of many biodegradable and non-degradable tubes with or without addition of molecules and cells. The main objective of the present study was to develop an economical and also an easy way for culturing a neural cell line which was capable of growing, differentiating and producing locally nerve growth factors that are otherwise extremely expensive, inside 90 PLA/10 PLG nerve guides. For this purpose the authors have chosen the N1E-115 cell line, a clone of cells derived from mouse neuroblastoma C-1300 with the perspective of using this differentiated cellular system to cover the inside of 90 PLA/10 PLG nerve guides placed to bridge a gap in the rat sciatic nerve experimental model. The N1E-115 cells proliferate in normal culture medium but undergo neuronal differentiation in response to DMSO. Upon induction of differentiation, proliferation of N1E-115 cells ceases, extensive neurite outgrowth is observed and the membranes become highly excitable. While it is known that Ca2+ serves as an important intracellular signal for various cellular processes, such as growth and differentiation. It is also known that can be toxic to cells and is involved in the triggering of events leading to excitotoxic cell death in neurons. The [Ca2+]i in non-differentiated N1E-115 cells and after distinct periods of differentiation, have been determined by the epifluorescence technique using the Fura-2-AM probe. The results of this quantitative assessment revealed that N1E-115 cells which undergo neuronal differentiation for 48 hours in the presence of 1.5% DMSO are best qualified to be used to cover the interior of the nerve guides since the [Ca2+]i was not found to be elevated indicating thus that the onset the cell death processes was not occurred. PMID- 16308462 TI - Modeling of chemical control of human respiratory system. AB - This paper addresses the fundamental understanding of the chemical control of respiratory system that would be needed to improve the efficiency of artificial ventilators and implement appropriate controllers. Hence, non-linear model for simulation of chemical control of human respiratory system under different physiological conditions is presented. The paper also presents the non-linear dissociation behaviors for oxygen and carbon dioxide solutions in the blood and the Bohr-Haldane effects as well as a new controller model. Tests were conducted on the model under different conditions such as hypoxia, hypercapnia, hypoventilation, and hyperventilation with some combination tests, in order to verify the validity of the assumptions that were made. Results indicate a very close agreement between the responses obtained from the present model and other published experimental and theoretical results. This model can be used to design a novel artificial respirator controller meeting the bodily requirements of the patient under mechanical ventilation. PMID- 16308463 TI - Effect of type of polymerization on different properties of dental composites. AB - The aim of this study is to assess the influence of plasma lamps on the properties of the composites compared to the influence of conventional polymerization. Vickers hardness tests, three-point bending tests, and measurement of the shrinkage marginal gap by scanning electron microscopy were carried out on three resin composites (Tetric Ceram, Z-100 and Inten-S) irradiated with to lamps (Flipo) plasma and Astralis 7 halogen lamps). With a 3 second exposure, the results of Vickers hardness and resistance to flexion (excepting values for Z-100) were lower for the composites cured by the Flipo plasma lamp, than after 40-second curing by the conventional halogen lamp (Astralis 7), notably at a depth of 3 mm. With a 5-second exposure the results of Vickers hardness and resistance to flexion obtained using the plasma lamp approached those obtained by using the halogen lamp. Whatever the polymerization protocol used, the measurements of the gap between the tooth and the filling are very similar except for Z-100/Astralis 7, for which shrinkage results are more important. For any one resin composite and lamp used, the shrinkage values obtained at a depth of 4 mm are twice higher than those obtained at the surface. In conclusion, for a 3-second exposure the level of polymerization obtained by plasma curing is lower than the one obtained by halogen curing, particularly in depth. On the other hand, 5-second plasma curing results recommends the use of this kind of lamp. PMID- 16308464 TI - Measuring cell viscoelastic properties using a force-spectrometer: influence of protein-cytoplasm interactions. AB - Cell adhesive and rheological properties play a very important role in cell transmigration through the endothelial barrier, in particular in the case of inflammation (leukocytes) or cancer metastasis (cancer cells). In order to characterize cell viscoelastic properties, we have designed a force spectrometer (AFM) which can stretch cells thereby allowing measurement of their rheological properties. This custom-made force spectrometer allows two different visualizations, one lateral and one from below. It allows investigation of the effects of rheology involved during cell stretching. To test the ability of our system to characterize such viscoelastic properties, ICAM-1 transfected CHO cells were analyzed. Two forms of ICAM-1 were tested; wild type ICAM-1, which can interact with the cytoskeleton, and a mutant form which lacks the cytoplasmic domain, and is unable to associate with the cytoskeleton. Stretching experiments carried out on these cells show the formation of long filaments. Using a previous model of filament elongation, we could determine the viscoelastic properties of a single cell. As expected, different viscoelastic components were found between the wild type and the mutant, which reveal that the presence of interactions between ICAM-1 and the cytoskeleton increases the stiffness of the cell. PMID- 16308465 TI - Interaction of fluorescent molecular rotors with blood plasma proteins. AB - Many disease states have associated blood viscosity changes. Molecular rotors, fluorescent molecules with viscosity sensitive quantum yields, have recently been investigated as a new method for biofluid viscosity measurement. Current viscometer measurements are complicated by proteins adhering to surfaces and forming air-surface layers. It is unknown at this time what effects proteins may have on biofluid viscosity measurements using molecular rotors. To answer this question, binding affinities to blood plasma proteins were investigated by equilibrium dialysis for four hydrophilic molecular rotors. Aqueous solutions of 9-[(2-cyano-2-hydroxy-carbonyl)vinyl]julolidine (CCVJ) and three derivatives were prepared and dialyzed against solutions of bovine source albumin, fibrinogen and immunoglobulin G approximating normal physiologic concentrations and fresh-frozen human plasma. After equilibration, dye concentration on each side of the dialysis membrane was assessed by spectrophotometry. The relative binding affinity of the four dyes to the proteins and to the plasma was compared. Affinity of all dyes was highest for albumin. The bound dye fraction showed little change in relation to protein concentration in the physiological concentration range. Diol, the most hydrophilic molecular rotor tested showed the lowest affinity for albumin. This study indicates that hydrophilic molecular rotors are well-suited for biofluid viscosity measurement. PMID- 16308466 TI - Morphology of elastase-induced cerebral aneurysm model in rabbit and rapid prototyping of elastomeric transparent replicas. AB - In this work, we describe a methodology to fabricate transparent elastomeric vascular replicas using rapid prototyping techniques. First, the three dimensional morphology of an elastase-induced aneurysm model in rabbit is acquired. The morphology is reconstructed from in vivo rotational angiography and it is compared with three-dimensional reconstructions obtained by computerized tomography and magnetic resonance imaging of an intraluminal arterial cast that was obtained from the same animal at sacrifice. Results show that resolution of the imaging modality strongly influences the level of detail, such as small side branches, in the final reconstruction. We developed an average morphology model for elastase-induced aneurysms in rabbits including the surrounding vasculature and describe a method for rapid prototyping of vascular models from the three dimensional morphology. Our replicas can be manufactured in a short period of time and the final product is optically clear. In addition, the elasticity of the models can be controlled to represent arterial elasticity, which makes them ideal for optical investigations of detailed flow dynamics using measurement tools such as particle image velocimetry. PMID- 16308467 TI - The distribution of rolling neutrophils in venular convergences. AB - The aim of this study was to characterize the distribution of adherent leukocytes in branched venular convergences in vivo. Intravital microscopy was used to obtain video images of leukocyte adhesion in multiple branched sites in mouse cremaster muscle, during the mild inflammatory response induced by surgical preparation. The average number of cells/vessel length was obtained over several minutes for seven venular convergences with varying geometrical configurations. Results from this study demonstrate a strong tendency of leukocytes to adhere at junctional points between converging vessels. Different vessel configurations were studied and results were shown to be insensitive to precise vessel geometry. Thus, in post-capillary venules, leukocytes are most likely to adhere at points between converging vessels, regardless of the precise geometrical properties or configuration of the vessels. Hydrodynamic mechanisms due to flow behavior through convergences likely play a significant role in determining locations of cellular adhesion. Future work should concentrate on quantifying the relative contributions of hydrodynamic and biochemical mechanisms to aid in understanding disease processes and development of treatments or therapeutics. PMID- 16308468 TI - Magnetic resonance microscopy determined velocity and hematocrit distributions in a Couette viscometer. AB - Magnetic resonance microscopy is used to non-invasively measure the radial velocity distribution in Couette flow of erythrocyte suspensions of varying aggregation behavior at a nominal shear rate of 2.20 s(-1) in a 1 mm gap. Suspensions of red blood cells in albumin-saline, plasma and 1.48% Dextran added plasma at average hematocrits near 0.40 are studied, providing a range of aggregation ability. The spatial distribution of the red blood cell volume fraction, hematocrit, is calculated from the velocity distribution. The hematocrit profiles provide direct measure of the thickness of the aggregation and shear rate dependent red blood cell depletion at the Couette surfaces. At the nominal shear rate studied hematocrit distributions for the red blood cells in plasma show a depletion zone near the inner Couette wall but not the outer wall. The red blood cells in plasma with Dextran show cell depletion regions of approximately 100 mum at both the inner and outer Couette surfaces, with greater depletion at the inner wall, but approach the normal blood hematocrit distribution with a doubling of shear rate due to decreased aggregation. The material response of the blood is spatially dependent with the shear rate and the hematocrit distribution non-uniform across the gap. PMID- 16308469 TI - The role of macrophages in tumor development. AB - Macrophages constitute a large proportion of the immune cell infiltrate, which is present in many tumors. Activation state of macrophages is greatly influenced by their environment, leading to different macrophage subsets with diverse functions. Although previously regarded as potent immune cells that are capable of destroying tumor cells, recent literature focuses on the ability of macrophages to promote tumor development due to secretion of mediators, like growth and angiogenic factors. It is now becoming increasingly clear that a complicated synergistic relationship exists between macrophages and malignant cells whereby tumor cells can affect macrophage phenotype, and vice versa. As such, macrophages and their contribution in cancer development are currently subject of debate. PMID- 16308470 TI - Identification of differentially expressed genes in metastatic and non-metastatic nasopharyngeal carcinoma cells by suppression subtractive hybridization. AB - BACKGROUND & OBJECTIVE: Nasopharyngeal carcinoma (NPC) is an epithelial neoplasm with high occurrence rates in southern China. The disease often metastasizes to regional lymphnodes at a very early stage. Local recurrences and metastasis occur frequently in patients with NPC and are a leading cause of death, despite improvements on treatment modalities. The molecular mechanism underlying the metastasis of nasopharyngeal carcinoma remains poorly understood, however, and requires additional elucidation. The aim of this study was to explore possible NPC gene candidates that may play key roles in NPC metastasis. METHODS: Subtractive suppression hybridization (SSH) was performed to isolate differentially expressed clones between the metastatic 5-8F and non-metastatic 6 10B nasopharyngeal carcinoma cell lines. Differentially expressed clones were screened and confirmed by reverse Northern blotting. The sequences of cDNA fragments were subsequently analyzed and compared to known sequences in Genbank. RESULTS & DISCUSSION: The SSH library contained thousands of positive clones. Random analysis of 300 clones by PCR demonstrated that 269 clones contained inserted fragments. Reverse Northern blot confirmed that 20 out of 192 clones examined were significantly up-regulated in the 5-8F cell line. Among these 20 clones, 16 were previously identified genes (flotilin-2, ezrin, pim-3, fli-1, mel, neugrin, znf216, ASB1, raly, UBE2A, keratin6A, TMED7, EIF3S9, FTL, two ribosomal proteins RPL21 and RPL16), two were predicted genes (c9orf74 and MDS006), and two sequences shared no homology with known genes listed in GenBank and may represent novel genes. The proposed functions of the genes identified in this study include cell signal transduction, cell survival, transcription regulation, cell mobility, protein synthesis, and DNA damage repair. Flotillin-2, fli-1, pim-3 and ezrin have previously been reported to be associated with tumor metastasis and progression. The remaining up-regulated genes identified in this study have not been reported to be markers of metastasis and may represent new candidates of NPC metastasis-related genes. The results of this study may provide novel points of therapeutic intervention for NPC. PMID- 16308471 TI - Implementation of accurate and fast DNA cytometry by confocal microscopy in 3D. AB - BACKGROUND: DNA cytometry is a powerful method for measuring genomic instability. Standard approaches that measure DNA content of isolated cells may induce selection bias and do not allow interpretation of genomic instability in the context of the tissue. Confocal Laser Scanning Microscopy (CLSM) provides the opportunity to perform 3D DNA content measurements on intact cells in thick histological sections. Because the technique is technically challenging and time consuming, only a small number of usually manually selected nuclei were analyzed in different studies, not allowing wide clinical evaluation. The aim of this study was to describe the conditions for accurate and fast 3D CLSM cytometry with a minimum of user interaction to arrive at sufficient throughput for pilot clinical applications. METHODS: Nuclear DNA was stained in 14 microm thick tissue sections of normal liver and adrenal stained with either YOYO-1 iodide or TO-PRO 3 iodide. Different pre-treatment strategies were evaluated: boiling in citrate buffer (pH 6.0) followed by RNase application for 1 or 18 hours, or hydrolysis. The image stacks obtained with CLSM at microscope magnifications of x40 or x100 were analyzed off-line using in-house developed software for semi-automated 3D fluorescence quantitation. To avoid sectioned nuclei, the top and bottom of the stacks were identified from ZX and YZ projections. As a measure of histogram quality, the coefficient of variation (CV) of the diploid peak was assessed. RESULTS: The lowest CV (10.3%) was achieved with a protocol without boiling, with 1 hour RNase treatment and TO-PRO-3 iodide staining, and a final image recording at x60 or x100 magnifications. A sample size of 300 nuclei was generally achievable. By filtering the set of automatically segmented nuclei based on volume, size and shape, followed by interactive removal of the few remaining faulty objects, a single measurement was completely analyzed in approximately 3 hours. CONCLUSIONS: The described methodology allows to obtain a largely unbiased sample of nuclei in thick tissue sections using 3D DNA cytometry by confocal laser scanning microscopy within an acceptable time frame for pilot clinical applications, and with a CV small enough to resolve smaller near diploid stemlines. This provides a suitable method for 3D DNA ploidy assessment of selected rare cells based on morphologic characteristics and of clinical samples that are too small to prepare adequate cell suspensions. PMID- 16308472 TI - Phenotypic changes in mitochondrial membrane potential (Delta psi(m)) during valinomycin-induced depolarisation and apoptosis. AB - A large body of evidence has implicated mitochondria in control of cell death, where key apoptotic mechanisms involve change in mitochondrial membrane permeability and depolarisation of mitochondrial membrane potential (Delta psi(m)). Assessment of Delta psi(m) is traditionally conducted using the lipophilic cation JC-1 on the flow cytometer or by fluorescent microscopy. Here we assess JC-1 aggregation using the novel tool of digital texture analysis to establish mitochondrial phenotypic changes induced by the K+ ionophore, valinomycin in a unique model comprising SW480 and SW620 cell lines. This provides an opportunity to study these phenomena in the context of colorectal cancer. Valinomycin-induced apoptosis was detected using morphology and analysis of DNA content. Cells were treated with valinomycin, images digitally recorded on a calibrated video photometer and subjected to high resolution digital texture analysis. This demonstrated that the HARAM texture features (Mean of the Haralick texture features) were highly valuable in describing the transition of Delta psi(m) as the cell undergoes apoptosis. In conclusion this study illustrates the potential of texture analysis as a novel and additional technique for quantifying JC-1 aggregation and revealing the spectrum of collapse of Delta psi(m) during apoptosis. PMID- 16308473 TI - Multi-resolution wavelet-transformed image analysis of histological sections of breast carcinomas. AB - Multi-resolution images of histological sections of breast cancer tissue were analyzed using texture features of Haar- and Daubechies transform wavelets. Tissue samples analyzed were from ductal regions of the breast and included benign ductal hyperplasia, ductal carcinoma in situ (DCIS), and invasive ductal carcinoma (CA). To assess the correlation between computerized image analysis and visual analysis by a pathologist, we created a two-step classification system based on feature extraction and classification. In the feature extraction step, we extracted texture features from wavelet-transformed images at 10x magnification. In the classification step, we applied two types of classifiers to the extracted features, namely a statistics-based multivariate (discriminant) analysis and a neural network. Using features from second-level Haar transform wavelet images in combination with discriminant analysis, we obtained classification accuracies of 96.67 and 87.78% for the training and testing set (90 images each), respectively. We conclude that the best classifier of carcinomas in histological sections of breast tissue are the texture features from the second-level Haar transform wavelet images used in a discriminant function. PMID- 16308474 TI - p53, Bcl-2 and C-Myc expressions in colorectal carcinoma associated with schistosomiasis in Egypt. AB - BACKGROUND AND AIMS: Oncogenes and tumor suppressor genes expression are well described in bladder cancer associated with schistosomiasis especially in Egypt. Scarce studies were directed to colorectal cancer (CRC) associated with Schistosoma mansoni (S. mansoni). Apoptosis (programmed cell death) and the genes regulating this process (e.g., Bcl-2) have recently become a focus of interest in the study of cancer development and progression. In the present study, we aimed to investigate the expression pattern of p53, Bcl-2 and C-Myc in CRC tissues obtained from Egyptian colorectal cancer patients divided in two different groups, one associated with Schistosoma mansoni (CRC-Sm) and the other without Schistosoma mansoni (CRC-NSm). METHODS: Seventy-five CRC tumors containing 36 draining lymph node metastatic tumors were immunohistochemically stained using specific monoclonal antibodies for p53, Bcl-2 and C-Myc, in addition the apoptotic activity of these tumors were analyzed. RESULTS AND CONCLUSIONS: Regardless of the S. mansoni infection, the obtained results showed that the apoptotic activity was more evident in p53 diffuse positive tumors (P = 0.021). There was a significant correlation between p53 diffuse positive staining and Bcl 2 positive immunostaining (P = 0.011). Signet ring cell carcinoma and mucinous adenocarcinoma exhibited both intense C-Myc expression than non-mucinous carcinoma (P = 0.001). When adjusting for S. mansoni infection, 58.3% of CRC-Sm cases were Bcl-2 positive compared to only (33.3%) of CRC-NSm (P = 0.046). Apoptotic activity was more evident in the latter group than of CRC-Sm tumors (P = 0.009). p53 and C-Myc expressions were found insignificantly different in CRC Sm compared with CRC-NSm (P > 0.05). These observations suggest that the genotoxic agents produced endogenously through the course of schistosomiasis mansoni may play a role in CRC-Sm pathogenesis through the dysregulation of apoptosis by alteration the expression pattern of Bcl-2 protein differently from CRC-NSm suggesting a different biological behavior. PMID- 16308475 TI - Digital image DNA cytometry: a useful tool for the evaluation of malignancy in biliary strictures. AB - BACKGROUND: Cytologic evaluation of the biliary tract strictures is nowadays widely used for distinguishing between benign and malignant lesions but remains a challenge for some problematic cases. Digital Image cytometry (DNA-cytometry) helps cytopathologists to resolve some unclear situations. METHODS: We have analysed 41 specimens of bile duct brushings obtained from patients during ERCP (11 benign cases, 7 suspicious for malignancy cases and 23 malignant cases) by DNA-cytometry and correlated them with the histological biopsy counterpart. RESULTS: All eleven cytological and histological benign cases were DNA-diploid and among 22 patients with malignant cytological and histological diagnosis 21 were DNA-aneuploid. One case considered malignant by the cytopathologist revealed DNA-aneuploid but malignancy could not be confirmed by histology. The analysis of the suspicious for malignancy cases revealed that all DNA-aneuploid cases were malignant and all DNA-diploid cases were benign referring to the follow-up of the patients. The comparison between cytology alone and cytology combined with DNA cytometry related to the histological diagnosis (gold standard) resulted in a sensitivity of 100% and a specificity of 79% for cytology alone; a specificity of 94% and a sensitivity 92% for DNA-cytometry and a specificity of 93% and a sensitivity of 100% with combined analyses. The positive predictive value was 90% for cytology, 96% for DNA-cytometry and for both analyses. The negative predictive value showed 100% for cytology, 89% for DNA-cytometry and 100% for combined studies. CONCLUSIONS: Despite the limited number of patients involved in the study, the results obtained indicate an increased of specificity and of positive predictive value using DNA-cytometry. These results confirm the pertinence of these method for challenging cases, in conjunction with other available diagnostic tools. PMID- 16308476 TI - Divergent autologous T cell responses to leukaemic dendritic cells during remission in acute promyelocytic leukaemia. PMID- 16308478 TI - Zinc dyshomeostasis: a key modulator of neuronal injury. AB - Zn2+ is a potently toxic cation involved in the neuronal injury observed in cerebral ischemia, epilepsy, and brain trauma. Toxic Zn2+ accumulation may result from either trans-synaptic Zn2+movement and/or cation mobilization from intracellular sites. To gain entry to the cytosol, Zn2+ can flux through glutamate receptor-associated channels, voltage-sensitive calcium channels, or Zn2+-sensitive membrane transporters, while metallothioneins and mitochondria provide sites of intracellular Zn2+ release. Intracellular Zn2+ homeostasis is sensitive to patho-physiological environmental changes, such as acidosis, inflammation and oxidative stress. The mechanisms by which Zn2+ exerts its neurotoxicity include mitochondrial and extra-mitochondrial production of reactive oxygen species and disruption of metabolic enzymatic activity, ultimately leading to activation of apoptotic and/or necrotic processes. Beside acute neuronal injury, an exciting new area of investigation is offered by the role of Zn2+ dysmetabolism in Alzheimer's disease as the cation acts as a potent trigger for Abeta aggregation and plaque formation. Finally, recent findings suggest that alteration of Zn2+ homeostasis might also be a critical contributor to aging-related neurodegenerative processes. Thus, multiple evidence suggest that modulation of intracellular and extracellular Zn2+ might be an important therapeutical target for the treatment of a vast array of neurological conditions ranging from stroke to Alzheimer's disease. PMID- 16308479 TI - Metallothionein-I-II and GFAP positivity in the brains from frontotemporal dementia patients. AB - Frontotemporal dementia regards a group of presenile progressive neurodegenerative form of dementias which includes Pick's disease, corticobasal degeneration, frontotemporal dementia with motor neuron disease, frontal lobe degeneration, dementia-parkinsonism-amyotrophy complex, familial non-specific dementia mapping to chromosome 3, non-Alzheimer degenerative dementia lacking distinctive histological features as well as a number other infrequent syndromes with dementia and focal neurological signs. The aim of this study was to investigate the regional distribution of metallothionein-I-II, an ubiquitary group of buffering proteins, in cases of frontotemporal dementia. The aim of the present study was to study the metallothionein-I-II expression in relationship to the expression in astrocytes of glial fibrillary acidic protein (GFAP) as we have already done in previous studies of Alzheimer's and Binswanger's diseases [31,32]. Our findings indicate that metallothionein-I-II expression in the most affected areas is likely to be regionally distinct and layer-dependent, in that it is highest in the deep layers of the frontotemporal cortex and the allocortex (hippocampus) while insignificantly immunopositive in the occipital cortex. In addition, the potential use of metallothionein-I-II as a new pharmacological approach to contrast some deleterious aspects of this disease has been also discussed. PMID- 16308480 TI - Synergistic effects of iron and aluminum on stress-related gene expression in primary human neural cells. AB - Disturbances in metal-ion transport, homeostasis, overload and metal ion-mediated catalysis are implicated in neurodegenerative conditions such as Alzheimer's disease (AD). The mechanisms of metal-ion induced disruption of genetic function, termed genotoxicity, are not well understood. In these experiments we examined the effects of non-apoptotic concentrations of magnesium-, iron- and aluminum sulfate on gene expression patterns in untransformed human neural (HN) cells in primary culture using high density DNA array profiling and Western immunoassay. Two week old HN cells were exposed to low micromolar magnesium, iron, or aluminum for 7 days, representing trace metal exposure over one-third of their lifespan. While total RNA yield and abundance were not significantly altered, both iron and aluminum were found to induce HSP27, COX-2, betaAPP and DAXX gene expression. Similarly up-regulated gene expression for these stress-sensing, pro-inflammatory and pro-apoptotic elements have been observed in AD brain. The combination of iron and aluminum together was found to be particularly effective in up regulating these genes, and was preceded by the evolution of reactive oxygen intermediates as measured by 2',7'-dichlorofluorescein diacetate assay. These data indicate that physiologically relevant amounts of iron and aluminum are capable of inducing Fenton chemistry-triggered gene expression programs that may support downstream pathogenic responses and brain cell dysfunction. PMID- 16308481 TI - Amyloid-beta: acute-phase apolipoprotein with metal-binding activity. AB - In monomeric form, amyloid-beta (Abeta), an amphipatic 4 kDa peptide which plays a key role in the pathogenesis of Alzheimer's disease (AD), is associated with lipoproteins in biological fluids and possesses high affinity to transition metal ions; in addition, synthesis of Abeta is upregulated under acute phase conditions. These data suggest that Abeta can be regarded as a normal physiological component of lipoproteins and may represent an acute-phase apolipoprotein with metal-binding activity. Interaction with transition metal ions may cause aggregation of Abeta, leading to formation of oligomers. Such oligomerised Abeta may lose its normal biological functions and acquire deleterious activities. To be efficient, novel anti-AD therapies should target oligomeric forms of Abeta while preserving the monomeric form of the peptide. PMID- 16308482 TI - The role of MT in neurological disorders. AB - Metallothioneins (MTs) are ubiquitous low molecular weight proteins characterized by their abundance of the thiol (SH)-containing amino acid, cysteine. To date four MT isoforms have been identified and cloned in mammals. MT-I and MT-II, the most widely expressed isoforms are generally coordinately regulated in all mammalian tissues; MT-III, is predominantly expressed in zinc (Zn)-containing neurons of the hippocampus; MT-IV is not expressed in brain tissue. The MT proteins have been implicated in gene expression regulation, homeostatic control of cellular metabolism of metals, and cellular adaptation to stress, including oxidative stress. MTs therefore impact on transcription, replication, protein synthesis, metabolism, and numerous other Zn-dependent biological processes. Disordered MT homeostasis leads to changes in brain concentrations of Zn. Since intracellular concentration of Zn are mediated by complexing with apothionein to form MT, there has been great interest in ascertaining whether disordered MT regulation plays a role in the etiology of neurodegenerative disorders. Though abnormalities in MT and/or Zn homeostasis have been reported in multiple neurological disorders a definitive link between MTs and the above disorders remains to be established. The chapter will commence with a brief discussion on the various MT isoforms, their structure and abundance (in brain), followed by a survey on the ability of MTs to potentiate or attenuate neurodegenerative process, with major emphasis on the role of MTs in the etiology of Alzheimer disease (AD). PMID- 16308483 TI - Calcium in Alzheimer's disease pathogenesis: too much, too little or in the wrong place? AB - Our understanding of the molecular genetics and biochemical pathology of Alzheimer's disease has progressed tremendously in the past decade. The metabolism of amyloid beta-peptide is being unraveled, and specific anti-amyloid therapies are now in clinical trials worldwide. The precise biophysical structure of the amyloid beta-peptide that causes neuronal dysfunction remains under investigation, as does the interaction between amyloid peptides and tau hyperphosphorylation, but these two molecules likely play key roles in neuronal dysfunction in Alzheimer's disease. Despite these advances, the cell biology of neuronal dysfunction and cell death in the Alzheimer's disease brain remains poorly understood. This brief review will explore the role of calcium (Ca2+) in neuronal death occurring during Alzheimer's disease. The evidence for glutamate receptor-mediated Ca;2+ overload, or excitotoxicity, and other derangements of Ca2+ homeostasis in cell culture and animal models of Alzheimer's disease is reviewed. Finally, we raise the possibility that some of the neuronal death observed in Alzheimer's disease might be associated with a reduction in rather than an increase in cytosolic Ca2+ levels, an idea with potentially important therapeutic implications. PMID- 16308484 TI - Is zinc the link between compromises of brain perfusion (excitotoxicity) and Alzheimer's disease? AB - Prior brain injury is a major risk factor in the development of Alzheimer's disease. This is true for traumatic brain injury, stroke or ischemic brain injury, and (more speculatively) for brain injury resulting from the hypo perfusion-reperfusion in cardiac arrest or cardiac bypass surgery and even hypo- or hypertension. Here we propose that the release of excess, toxic, "floods" of free zinc into the brain that occurs during and after all excitotoxic brain injury is a key factor that sets the stage for the later development of Alzheimer's disease. Rapid and aggressive administration of zinc buffering compounds to patients suffering brain injury may therefore not only ameliorate the acute injury but might also reduce the risk of subsequent development of Alzheimer's disease. PMID- 16308485 TI - Imbalance between pro-oxidant and pro-antioxidant functions of zinc in disease. AB - Alzheimer's disease is associated with oxidative stress and changes in metal metabolism. Among the essential trace metals, zinc has the greatest number and variety of functions in hundreds of enzymes and thousands of protein domains with different types of zinc finger motifs. Moreover, zinc ions are stored in synaptic vesicles of specialized neurons and released during neuronal activity. Based on this multitude of functions, one would expect that impairment of zinc homeostasis in the brain has far-reaching consequences. In spite of the fact that zinc ions are redox-inert in biology, they have profound effects on redox metabolism. Thus, both zinc deficiency and zinc overload elicit oxidative stress that can lead to the death of nerve cells. These pro-oxidant functions contrast with pro antioxidant functions in a range of physiological zinc concentrations. Oxidative or nitrosative stress can release zinc from proteins with zinc finger and cluster motifs and re-distribute zinc, thereby changing the functions of the proteins from which it is released and to which it binds. The transduction of redox signals into zinc signals and vice versa affects mitochondrial functions and signaling pathways (NF-kappaB, p53, AP-1) where zinc and the zinc donor/acceptor pair metallothionein/thionein are critically involved in life and death decisions of the cell. PMID- 16308486 TI - Effects of aluminum on the nervous system and its possible link with neurodegenerative diseases. AB - Aluminum is environmentally abundant, but not an essential element. Aluminum has been associated with several neurodegenerative diseases, such as dialysis encephalopathy, amyotrophic lateral sclerosis and Parkinsonism dementia in the Kii peninsula and Guam, and in particular, Alzheimer's disease. Although this association remains controversial, there is increasing evidence which suggests the implication of metal homeostasis in the pathogenesis of Alzheimer's disease. Aluminum, zinc, copper, and iron cause the conformational changes of Alzheimer's amyloid-beta protein. Al causes the accumulation of tau protein and amyloid-beta protein in experimental animals. Aluminum induces neuronal apoptosis in vivo as well as in vitro. Furthermore, a relationship between aluminum and the iron homeostasis or calcium-homeostasis has been suggested. Based on these findings, the characteristics of aluminum neurotoxicity are reviewed, and the potential link between aluminum and neurodegenerative diseases is reconsidered. PMID- 16308487 TI - Iron, neuroinflammation, and Alzheimer's disease. AB - The present article reviews the roles and interactions of iron and neuroinflammation in Alzheimer's disease. It highlights the importance of neuroinflammatory changes in the glial scar after neuronal injury, in promoting iron accumulation, and iron-dependent oxidative damage. Elevation of iron in Alzheimer's disease not only affects amyloid precursor protein processing and mitochondrial function but also induces the aggregation of Abeta peptide and abnormalities in signal transduction processes associated with oxidative damage. Collective evidence suggests that although alterations in iron homeostasis may not be the primary triggering event that starts the pathological cascade of Alzheimer's disease, it is an important factor involved in neuroinflammation and progression of this disease. PMID- 16308488 TI - Involvement of amyloid beta precursor protein (AbetaPP) modulated copper homeostasis in Alzheimer's disease. AB - AbetaPP is involved in Cu homeostasis in mouse and man. In vitro observations and in vivo data obtained from AbetaPP mouse models at least provide strong evidence that AbetaPP and Abeta overproduction enables intracellular Cu to be transported out of the cell. The increased Cu efflux seems to lead to a Cu deficiency and a subsequently reduced SOD-1 activity. Studies have shown that a disturbed metal ion homeostasis with elevated serum Cu levels occurs in Alzheimer and Down's patients and lowered levels in post-mortem AD brain. We conclude that bioavailable Cu has beneficial and specific effects in Alzheimer's disease transgenic mice, and suggest that our observation can be regarded as a proof-of concept for a prophylactic approach to overcome the observed CNS Cu deficiency in the brain of Alzheimer's disease patients. PMID- 16308489 TI - Progress in myocardial regeneration and cell transplantation. AB - Embryonic stem cells and bone marrow mesenchymal stem cells can be induced to differentiate into cardiomyocytes. Techniques to purify and transplant regenerated cardiomyocytes have been developed, and transplanted regenerated cardiomyocytes are capable of residing in the heart of recipients for long periods. Advances in tissue engineering technology have enabled the production of cardiomyocyte cell sheets for transplantation treatment of heart failure, without the need for a donor, and this has now reached the preclinical stage. The treatment of heart failure using cytokines to mobilize stem cells has also been explored. PMID- 16308490 TI - Disparity in the process and outcome of the treatment for acute myocardial infarction in Japan: CAMPAIGN Study in the National Hospital Network. AB - BACKGROUND: A nationwide survey of the process and outcome of treatment for acute myocardial infarction (AMI) has not been conducted in Japan. METHODS AND RESULTS: In the present study 2,007 patients with AMI admitted to 22 national hospitals were registered between July 1999 and January 2002 for CAMPAIGN Study 1; an additional 206 and 238 cases were registered between October and December 2002 (CAMPAIGN 2) and between October and December 2003 (CAMPAIGN 3), respectively. In CAMPAIGN 1, the length of stay varied from 15 to 35 days among hospitals (mean: 24.8 days), and was mainly determined by the schedule of follow-up examinations rather than clinical course. Of the prescriptions at discharge, beta-blockers and angiotensin-converting enzyme inhibitors varied widely; the use of beta-blockers was very low (25%). Nitrates were frequently used (68%) although there is no evidence for secondary prevention. In CAMPAIGNs 2 and 3, the use of beta-blockers increased (36%, 47%) and that of nitrates decreased (24%, 21%). CONCLUSION: CAMPAIGN Study 1 revealed considerable variation in the treatment of AMI during the acute phase among the hospitals. The use of beta-blocker and nitrates as discharge medication was inappropriate. CAMPAIGNs 2 and 3 showed some improvement in the problems revealed by CAMPAIGN 1. PMID- 16308491 TI - Acute myocardial infarction in young Japanese adults. AB - BACKGROUND: The prevalence and clinical manifestations of acute myocardial infarction (AMI) in young patients in Japan have not been fully investigated. METHODS AND RESULTS: In the present study, 1651 AMI patients were enrolled in the AMI-Kyoto Multi-Center Risk Study between January 2000 and June 2004. Of these, the clinical background, risk factors, angiographic findings, acute results of primary percutaneous coronary intervention (PCI) and in-hospital outcomes for 27 young patients <40 (young group), and 338 non-young patients 60or=0.5) and poorly fitted regions (R2<0.5). Regional relative myocardial blood flow calculated from the initial distribution of 11C-acetate and left ventricular (LV) wall motion were also evaluated. Peak 11C-palmitate uptake (14,434+/-3,052 vs 12,016+/-3,088 counts/s, p<0.001) and percent clearance during acquisition (38.2+/-10.1 vs 23.6+/-11.4%, p<0.001) were significantly greater in the well fitted regions (n=111) than in the poorly fitted regions (n=44). Acetate-Kmono was significantly higher in the former than in the latter (0.0641+/-0.0099 vs 0.0476+/-0.0103 min 1, p<0.001). LV wall motion and regional relative blood flow were also significantly greater in the former regions. Palmitate-Kmono in the well fitted regions was significantly higher in normal LV wall motion areas than in hypokinesis areas (0.0363+/-0.0062 vs 0.0274+/-0.0057 min-1, p<0.001) CONCLUSIONS: Maintenance of myocardial fatty acid beta-oxidation with better myocardial blood flow is substantial in the preservation of total myocardial oxidative metabolism and LV wall motion in patients with MI. The finding that the early-phase clearance of 11C-palmitate is fitted with a monoexponential curve may provide important information in the evaluation of myocardial fatty acid beta oxidation. PMID- 16308493 TI - Vasospastic angina and microvascular angina are differentially influenced by PON1 A632G polymorphism in the Japanese. AB - BACKGROUND: Ethnicity and smoking are well-known risk factors for the pathogenesis of coronary vasospasm. Oxidative stress induced by smoking plays a crucial role in coronary vasospasm, but is not enough to account for the pathogenesis of coronary vasospasm, indicating that genetic factors are strongly involved. METHODS AND RESULTS: The study group comprised 162 vasospastic angina patients (VSAs), 61 microvascular angina patients (MVAs) and 61 non-responders (NRs) diagnosed by acetylcholine provocation test. Four polymorphisms of the oxidative stress related genes, cytochrome b-245, alpha polypeptide gene (CYBA) C242T and A640G, paraoxonase 1 gene (PON1) A632G, phospholipase A2 group VII gene (PLA2G7) G994T were genotyped. Allele frequency of PON1 632-G was significantly higher in both the VSA with dominant fashion and the MVA with recessive fashion compared with NR. This association was strongly influenced by gender in the MVA only. There were no significant associations between the other polymorphisms and coronary vasospasm. In addition, the allele frequency of PON1 632-G in the Japanese was higher than in Caucasians. CONCLUSIONS: There was a significant association between PON1 A632G polymorphism and MVA as well as VSA, but the impact of this on VSA and MVA is different in the Japanese. PMID- 16308494 TI - N-terminal pro-B-type natriuretic Peptide predicts significant coronary artery lesion in the unstable angina patients with normal electrocardiogram, echocardiogram, and cardiac enzymes. AB - BACKGROUND: Brain natriuretic peptide (BNP) and N-terminal pro-brain natriuretic peptide (NT-proBNP) are not specific for ventricular dysfunction and other cardiac processes, such as myocardial ischemia, may also cause elevation of these markers. METHODS AND RESULTS: To determine whether elevation of NT-proBNP without elevation of cardiac specific markers can predict coronary artery disease (CAD), the serum level of NT-proBNP was measured in 161 patients with unstable angina (61.0+/-8.1 years, male 54.0%) with normal ventricular function (left ventricular ejection fraction >55% and no regional wall motion abnormality by echocardiography) and normal troponin I level (<0.05 ng/ml). In these patients, levels of C-reactive protein and myoglobin were normal and none had Q wave on electrocardiographic (ECG). The NT-proBNP level was higher in patients with CAD (n=74) than in patients without CAD (n=87) (173.1+/-231.6 vs 68.1+/-62.5 pg/ml, p<0.001). At the standard cut-off point of >200 pg/ml, elevated NT-proBNP level shows high probability of CAD (odds ratio, 10.1; 95% confidence interval, 2.6 38.7, p=0.001). The NT-proBNP level positively correlated with the extent of CAD (r=0.329, p=0.001). In multivariate analysis, the NT-proBNP was an independent predictor of CAD. CONCLUSION: These results suggested that NT-proBNP is a useful screening test for CAD in the unstable angina patients with normal ECG, echocardiogram and cardiac enzyme levels. PMID- 16308495 TI - Elevated preprocedural high-sensitivity C-reactive protein levels are associated with neointimal hyperplasia and restenosis development after successful coronary artery stenting. AB - BACKGROUND: Recent data indicate that an elevated serum level of high-sensitivity C-reactive protein (hs-CRP) predicts the risk of recurrent coronary events, and that statin therapy decreases the risk of coronary events. This study assessed the relationship between the pre-procedural hs-CRP level and in-stent neointimal hyperplasia (NIH) after stenting and the effects of statins on the relationship between restenosis after stenting and the serum hs-CRP levels of patients with coronary artery disease. METHODS AND RESULTS: This study included 100 patients who underwent stent implantation for angiographically significant stenosis. Patients were divided into a normal C-reactive protein (CRP) group (<0.5 mg/dl, n=59) and elevated CRP group (>or=0.5 mg/dl, n=41). All patients underwent angiographic and intravascular ultrasound follow-up at 6 months. The baseline CRP level was 0.29+/-0.08 mg/dl in the normal CRP group and 2.90+/-2.31 mg/dl in the elevated CRP group. The NIH cross-sectional area (CSA) in the minimal lumen CSA at follow-up was significantly larger in the elevated CRP group compared with the normal CRP group (1.9+/-1.3 mm2 vs 3.0+/-1.5 mm2, p=0.001). A significant positive correlation was found between pre-interventional CRP level and NIH area (r=0.52, p<0.001). In patients with normal CRP, an association between statin therapy and restenosis was not observed. However, when the analysis was confined to patients with elevated CRP, statin therapy significantly reduced the restenosis rate (20% vs 37.5%, p=0.031). In the normal CRP group, the intra-stent neointimal area at 6 months was not different between the non-statin and statin groups (2.2+/-1.4 mm2 vs 1.8+/-1.1 mm2). However, in the elevated CRP group, statin therapy significantly decreased the neointimal area at 6-month follow-up (3.6+/-1.7 mm2 vs 2.4+/-1.3 mm2, p<0.001). CONCLUSION: Measuring the pre interventional hs-CRP level may help predict the development of restenosis after stenting and statin therapy will significantly reduce the restenosis rate in patients with an elevated hs-CRP. PMID- 16308496 TI - Elevated monocyte chemoattractant protein-1 serum levels in patients at risk for coronary artery disease. AB - BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) is involved in the recruitment of monocytes into the arterial vessel wall as one of the major events leading to atherosclerotic vascular diseases, such as coronary artery disease (CAD). METHODS AND RESULTS: The study group comprised 263 volunteers aged between 18 and 85 years who were admitted to hospital or clinic for scheduled invasive and non-invasive diagnostic procedures. MCP-1 serum levels were determined using a sandwich-enzyme-linked immunosorbent assay. In each patient, the coronary risk factors (CRF), such as hypertension, high cholesterol, diabetes mellitus, obesity, positive family history, and smoking were evaluated. Low-density lipoprotein-cholesterol, lipoprotein(a), and hemoglobinA1C levels were determined. Patients with CAD proven by angiography had significantly increased MCP-1 levels. In patients without CAD, the increase in MCP-1 depended on the number of CRF. As a marker for endothelial activation the soluble adhesion molecules, soluble intercellular adhesion molecule and soluble E-selectin were measured and both markers were significantly elevated in patients with CAD or multiple CRF when compared with patients without CRF. Although this is not a direct proof, endothelial activation could contribute to elevated MCP-1 levels in atherosclerosis. CONCLUSION: Elevated MCP-1 serum levels could serve as a direct marker of the inflammatory activity in patients at risk for coronary artery and other atherosclerotic vascular diseases. PMID- 16308497 TI - Sustained upregulation of inflammatory chemokine and its receptor in aneurysmal and occlusive atherosclerotic disease: results form tissue analysis with cDNA macroarray and real-time reverse transcriptional polymerase chain reaction methods. AB - BACKGROUND: Although cytokines are known to be pivotal in the development of atherosclerotic diseases, few data exist regarding their expressions in the established stages such as aneurysmal or occlusive lesions. Therefore, in the present study the gene expression levels of cytokine-related substances in abdominal aortic aneurysm (AAA) and carotid artery stenosis (CAS) were determined using cDNA macroarray and real-time reverse transcriptase polymerase chain reaction (RT-PCR) methods. METHODS AND RESULTS: Tissue samples were obtained from 31 patients with AAA and 24 with CAS. The array-specific [33P]-labeled cDNA probe mixture synthesized from 2.5 microg total RNA with gene-specific primers was hybridized with nylon membranes containing 375 cDNA clones. Densitometric analysis confirmed differences in expression (>5-fold) for 97 of the cytokine related gene products between AAA and adjacent control tissue. Among these, simultaneous upregulation was found in the expression of interleukin (IL)-8 (9 fold) and its receptor, CXCR-2 (11-fold). Thus, the expressions of IL-8 and CXCR 2 were further quantified by real-time RT-PCR. The expression of both the genes was significantly upregulated in both AAA and CAS compared with control regions as followed: IL-8=0.53+/-0.16 vs 0.11+/-0.04 (p<0.01); CXCR-2=2.04+/-0.75 vs 0.29+/-0.10 (p<0.01) in AAA, and IL-8=1.35 +/-0.25 vs 0.60+/-0.16; CXCR-2=2.00 +/ 0.51 vs 0.58+/-0.21 (p<0.05) in CAS. Under these conditions, the gene expressions of monocyte chemotactic protein-1 and its receptor, CCR-2, were not significantly different in the control and diseased regions of both AAA and CAS. CONCLUSIONS: Sustained upregulation of IL-8 and CXCR-2 was observed in both AAA and CAS, suggesting the inflammatory process is still active in established dilated and occlusive atherosclerotic diseases. Whether upregulation of this system could be protective or not protective for disease development requires further study. PMID- 16308498 TI - Comparison of 2 mapping strategies for pulmonary vein isolation. AB - BACKGROUND: Pulmonary vein (PV) isolation using a circular catheter (CC) creates an entrance block from the left atrium (LA) to PV to eliminate paroxysmal atrial fibrillation (PAF). We describe a new approach for PV isolation during distal PV pacing using a basket catheter (BC). The purpose of the present study was to compare 2 mapping strategies for PV isolation. METHODS AND RESULTS: Of 100 consecutive patients with PAF, 50 underwent CC-guided PV isolation and 50 underwent BC-guided PV isolation. During CC-guided ablation, the endpoint was the elimination of PV potentials based on the entrance block from the LA to PV. During BC-guided ablation during distal PV pacing, the endpoint was the elimination of bidirectional PV-LA conduction. At 12 months, 62% of patients who underwent CC ablation and 80% of patients who underwent BC ablation were free of symptomatic PAF without the use of antiarrhythmic drugs (p<0.05). The incidence of mild (<50%) PV stenosis in BC ablation was significantly lower than that in CC ablation (12 vs 24%, p<0.01). CONCLUSIONS: This new approach for PV isolation during distal PV pacing using BC is useful for confirming a bidirectional PV-LA conduction block and is more effective than CC ablation. PMID- 16308499 TI - Hybrid therapy with pilsicainide and pulmonary vein isolation for atrial fibrillation. AB - BACKGROUND: Pulmonary vein (PV) isolation is commonly performed in patients with drug-refractory atrial fibrillation (AF) and in cases of unsuccessful ablation, antiarrhythmic agents that were ineffective before the ablation sometimes become effective afterward. Therefore, the effects and mechanisms of hybrid therapy with pilsicainide and PV isolation for AF were assessed in the present study. METHODS AND RESULTS: Seventy-four patients with paroxysmal AF in whom pilsicainide was ineffective underwent PV isolation. If AF recurred, a second procedure was performed and if AF recurred again, pilsicainide was re-administered. After the first procedure, AF recurred in 42 patients, and a second procedure was performed in 31 patients, of whom 28 had recovery of left atrial (LA)-PV conduction and non PV foci were identified in 3. After the second session, ablation eliminated AF without drugs in 53 (72%) patients. Pilsicainide suppressed the conduction properties at the LA-PV junction. In 21 patients with recurrence of AF, pilsicainide was re-administered and eliminated AF in 11 patients (success with pilsicainide: 86%). CONCLUSIONS: In cases of unsuccessful PV isolation, pilsicainide may prevent AF by modifying the LA-PV conduction properties. Hybrid therapy with pilsicainide and PV isolation may be an effective therapeutic approach for AF. PMID- 16308500 TI - Nifekalant hydrochloride suppresses severe electrical storm in patients with malignant ventricular tachyarrhythmias. AB - BACKGROUND: Some patients with an implantable cardioverter-defibrillator (ICD) suffer from burst of inappropriate multiple discharges (severe electrical storm), and because the current therapeutic options are limited, the effect of nifekalant hydrochloride, a new class III drug, on severe electrical storm was investigated in the present study. METHODS AND RESULTS: Ninety-one consecutive patients treated with ICD were included in the study (M 70; mean age 58 years; left ventricular ejection fraction 45+/-15%). Severe electrical storm was defined as more than 10 ICD discharges within 1 h. During a mean follow-up period of 30+/-13 months, 41/91 (45%) patients had appropriate ICD therapy for arrhythmias and severe electrical storm occurred in 11 of them (12%) at 20+/-18 months after ICD implantation. The mean number of ICD discharges/h during severe electrical storm was 18+/-12. In 4 of 10 patients, severe electrical storm was successfully suppressed by a combination of deep sedation and beta-blocking agent; 6 other patients were refractory to this treatment, but severe electrical storm was successfully suppressed by intravenous administration of nifekalant hydrochloride with no adverse effects. CONCLUSIONS: Nifekalant hydrochloride is an effective and safe treatment for severe electrical storm. PMID- 16308501 TI - Recurrence of atrial fibrillation after internal cardioversion of persistent atrial fibrillation: prognostic importance of electrophysiologic parameters. AB - BACKGROUND: The purpose of this study was to determine whether the extent of atrial electrical remodeling affects the recurrence of atrial fibrillation (AF) after cardioversion of persistent AF (PAF). METHODS AND RESULTS: Internal atrial cardioversion was performed in 47 patients with PAF. The right atrial monophasic action potential duration (RA-MAPD) at pacing cycle lengths (PCLs) of 800-300 ms and P wave signal-averaged electrocardiogram were recorded after cardioversion. Bepridil (150-200 mg/day) and carvedilol (10 mg/day) were administered to all patients after cardioversion. Of the 47 patients, 20 had recurrent AF within 3 months. No relation was observed between age, left atrial dimension, left ventricular ejection fraction, and AF recurrence. The AF duration was significantly longer (p<0.05) and RA-MAPD at PCLs of 800 to 300 ms were significantly shorter (p<0.05) in patients with AF recurrence than in those without recurrence. The mean slope of the RA-MAPD for PCLs between 600 and 300 ms did not differ between the patients with and without AF recurrence. The filtered P-wave duration (FPD) was significantly longer in the patients with AF recurrence than in those without (p<0.05). Multivariate analysis also showed that the RA MAPD at a PCL of 300 ms and FPD were predictors of AF recurrence (RAMAPD: p=0.038; FPD: p=0.052). CONCLUSION: These results suggest that electrical remodeling related to the repolarization and depolarization may be the main contributors to early AF recurrence after cardioversion under the administration of bepridil and carvedilol. PMID- 16308502 TI - Efficacy of the phase images in Fourier analysis using gated cardiac POOL-SPECT for determining the indication for cardiac resynchronization therapy. AB - BACKGROUND: Although cardiac resynchronization therapy (CRT) improves quality of life and survival for patients with heart failure, exact methods to estimate the effect of cardiac asynchrony have not yet been defined. METHODS AND RESULTS: Initially, to examine whether the phase analysis images in the Fourier analysis using gated cardiac pool single photon emission computed tomography (POOL-SPECT) could be used to evaluate cardiac asynchrony, 19 consecutive patients with dilated cardiomyopathy were studied. Interventricular asynchrony was defined by whether the peak of the picture elements of the right ventricle in the phase histogram fitted that of the left ventricle and intraventricular asynchrony by whether the phase image was described homogenously or not. The patients with both inter- and intraventricular asynchrony had significant deterioration in both left ventricular ejection fraction (p<0.01) and New York Heart Association functional class (p<0.01). To evaluate the efficacy of these phase images for CRT setting, 7 patients were tested before and after CRT. During a 3.9+/-3.6 month follow-up period, all patients had an improvement in their condition, and the inter- and intraventricular asynchrony significantly improved after CRT. The degrees of the inter- and intraventricular asynchrony were related to the degree of cardiac depression pre CRT. CONCLUSION: These results have shown that the phase images from POOL-SPECT are useful for assessing the effect of CRT in patients with heart failure, which suggests that it may provide information about the indication for CRT. PMID- 16308503 TI - Assessment of quality of life with 5 different scales in patients participating in comprehensive cardiac rehabilitation after acute myocardial infarction. AB - BACKGROUND: Measures assessing quality of life (QOL) in patients participating in comprehensive cardiac rehabilitation (CCR) have not been established in Japan. METHODS AND RESULTS: To compare different types of QOL scales and to determine the impact of CCR on QOL in Japanese cardiac patients, 5 different types of questionnaires were assessed in 44 patients participating in CCR after acute myocardial infarction (AMI). After 3-month CCR, peak oxygen uptake (PVO2, p<0.01), Sickness Impact Profile (SIP) total score (p<0.05) and physical function related QOL scores (Specific Activity Scale (SAS), p<0.01; SIP physical score, p<0.01) significantly improved, whereas psychosocial/mental aspect-related QOL scores (Ministry of Health and Welfare (MHW)-QOL score, SIP psychosocial score, State-Trait Anxiety Inventory, Self-rating Depression Scale) did not change on the average. However, patients with low PVO2 (<21.7 ml.min-1.kg-1) showed significant improvements in all scores after CCR, whereas patients with preserved exercise capacity showed improvements only in physical function-related scores (SAS and physical SIP). Furthermore, patients with anxiety and depression showed significant improvements in these respective measures after CCR. CONCLUSION: In patients with AMI, physical function-related QOL scores improve after a 3-month CCR program, but psychosocial/mental aspect-related QOL scores improve only in those with impaired exercise tolerance or anxiety/depression. Thus, changes in QOL after CCR depend on type of QOL scale used and the baseline status of the patient. In addition, in Japanese cardiac patients MHW-QOL mainly reflects psychosocial/mental aspect-related QOL, as well as overall QOL. PMID- 16308504 TI - Accelerated progression and morbidity in patients with aortic stenosis on chronic dialysis. AB - BACKGROUND: Progression of aortic stenosis (AS) influences the prognosis in chronic dialysis (CD) patients, but there have been few studies in Japanese patient populations. METHODS AND RESULTS: The progression of AS was examined in 16 CD patients (68+/-10 years) and 82 non-dialysis patients (73+/-11 years). The mean follow-up was 32 and 40 months, respectively. The calcium-phosphate product was increased in CD patients (49+/-14 mg/dl vs 30+/-5 mg/dl; p<0.0001). At entry, the peak transaortic gradient detected by Doppler echocardiography was lower in CD patients (42+/-12 mmHg vs 57+/-22 mmHg; p<0.05). At follow-up, there was no difference between the 2 groups in that value (69+/-31 mmHg vs 71+/-27 mmHg). Decreasing rate of calculated aortic valvular area was increased in CD patients (0.14+/-0.13 cm2/year vs 0.06+/-0.09 cm2/year; p<0.05) and the calcification of the aortic valve was more severe at follow up in CD patients. The mortality was higher in CD patients (75 vs 28%; p<0.001). CONCLUSIONS: AS appeared to progress rapidly in CD patients and therefore early aortic valve replacement may be necessary. PMID- 16308505 TI - Plasma adenosine levels in peri-menopausal women having frequent hot flushes. AB - BACKGROUND: The effect of hormone replacement therapy (HRT) on plasma adenosine levels was investigated in climacteric women experiencing hot flushes. METHODS AND RESULTS: Plasma adenosine levels were measured in 13 peri-menopausal women with frequent hot flushes (>5 per day) before and 3-4 months after initiating HRT. Thirteen healthy pre-menopausal, 9 peri-menopausal women with few hot flushes (<1 or 2 per day) and 10 healthy postmenopausal women were enrolled as controls. The average plasma adenosine level in the peri-menopausal women was 0.20+/-0.09 micromol/L, which was significantly higher than in the pre-menopausal (0.12+/-0.07 micromol/L, p<0.05), peri-menopausal with few hot flushes (0.10+/ 0.09 micromol/L, p<0.05) and postmenopausal women (0.13+/-0.06 micromol/L, p<0.05). Both the increased adenosine level and the hot flushes were decreased by HRT (plasma adenosine: 0.13+/-0.06 micromol/L). CONCLUSIONS: Increased plasma adenosine in peri-menopausal women may be associated with frequent hot flushes. PMID- 16308506 TI - Effects of late administration of immunoglobulin on experimental atherosclerosis in apolipoprotein E-deficient mice. AB - BACKGROUND: Although immunoglobulin treatment, beginning simultaneously with the initiation of atherosclerosis, suppresses experimental atherosclerosis in apolipoprotein E-deficient mice, it remains unclear whether the treatment at a subsequent stage of atherosclerosis would be effective. METHODS AND RESULTS: Experimental atherosclerosis was induced in mice fed a high-fat diet containing 0.3% cholesterol. After confirming the presence of atherosclerotic lesions at 11 weeks, the mice were treated with an intraperitoneal injection of either intact type of immunoglobulin or F(ab')2 fragments of immunoglobulin (both, 1 g.kg-1.day 1) on alternate days over 4 weeks. Fatty streak lesion was suppressed by intact immunoglobulin administration, but not by F(ab')2 fragments of immunoglobulin. Immunohistochemical analysis showed that macrophage and CD4+ T-cell accumulation in the fatty streak lesion was suppressed in mice that received intact immunoglobulin but not in those that received F(ab')2 fragments. CONCLUSIONS: Immunoglobulin treatment, even at a later stage of atherosclerosis, suppresses the development of lesions associated with the reduced expression of immune activated cells in fatty streak plaques, demonstrating the benefits of immunoglobulin therapy for prevention of atherosclerosis. PMID- 16308507 TI - Biphasic effect of HMG-CoA reductase inhibitor, pitavastatin, on vascular endothelial cells and angiogenesis. AB - BACKGROUND: HMG-CoA reductase inhibitors (statins) have pleiotropic effects beyond their cholesterol-lowering effect. However, consensus on the effect of statins on endothelial cells and angiogenesis has not yet been reached. METHODS AND RESULTS: The effects of pitavastatin on the migration, proliferation and viability of human epidermal microvessel endothelial cells (HMVECs) were examined using scratch assay, chemotaxis chamber, bromodeoxyuridine incorporation, trypan blue dye exclusion test, and nuclear DNA staining. Pitavastatin enhanced the migration, proliferation and viability of HMVECs at a low concentration (0.01 micromol/L) but inhibited them at high concentration (1 micromol/L). The inhibitory effect on cell viability by high concentration of pitavastatin was recovered by geranylgeranyl pyrophosphate, but the effect on migration and proliferation was not. The cell activating effect of a low concentration of pitavastatin was reversed by both farnesyl pyrophosphate and geranylgeranyl pyrophosphate. A quail chorioallantoic membrane assay showed that high concentration (1 micromol/L) of pitavastatin reduced fibroblast growth factor-2 induced angiogenesis, whereas low concentration (0.3 micromol/L) tended to increase angiogenesis. CONCLUSION: Pitavastatin has a biphasic effect on HMVECs and on angiogenesis through at least 2 different pathways that include the mevalonate pathway. PMID- 16308509 TI - Congenital mitral insufficiency caused by anomalous mitral arcade in an elderly patient: use of echocardiography and multidetector computed tomography for diagnosis. AB - Anomalous mitral arcade is a rare congenital malformation and few survive to adulthood. A 65-year-old man presented with mild dyspnea and cardiac murmur. Transthoracic and transesophageal echocardiography showed an arc-like configuration of the papillary muscles, constrained by an interconnecting fibrous band in direct continuity with the anterior mitral leaflet, and mitral insufficiency caused by the restricted mobility of the mitral valve. Multidetector computed tomography confirmed these findings. PMID- 16308508 TI - Usefulness of delayed enhancement magnetic resonance imaging for detecting cardiac rupture caused by small myocardial infarction in a case of cardiac tamponade. AB - Delayed enhancement magnetic resonance imaging (DE-MRI) has excellent spatial resolution and compared with other cardiac imaging techniques it can detect a small myocardial infarction (MI) or a subendocardial infarction. A 76-year-old man was admitted for loss of consciousness because of cardiac tamponade. The cause of tamponade was unknown, but electrocardiography and blood test suggested a recent MI. The removal of 100 ml bloody fluid by immediate pericardiocentesis normalized his hemodynamics, and he regained consciousness. Neither echocardiography nor scintigraphy could determine the location of the MI or rupture, but DE-MRI clearly demonstrated a transmural enhancement in a very narrow range of the lateral wall of the left ventricle. Coronary angiography revealed a severely stenotic lesion in the obtuse marginal branch of the left circumflex artery. DE-MRI is a powerful tool for diagnosing small MI that are undetectable with other imaging. Therefore, DE-MRI should be applied in cases with cardiac tamponade by unknown causes. PMID- 16308510 TI - Acute myocardial infarction caused by "malignant" anomalous right coronary artery detected by multidetector row computed tomography. AB - Anomalous coronary arteries are usually identified incidentally by angiography or autopsy, but some "malignant" coronary anomalies are associated with a high incidence of syncope, arrhythmia, myocardial infarction, and sudden death. So far, the pathogenesis of the coronary events in such cases has only been revealed by autopsy. In the present case report, a patient with anomalous origin of the right coronary artery from the left sinus of Valsalva developed acute myocardial infarction, and visualization of the anomaly and assessment of the culprit plaque in the artery were done by multidetector row computed tomography and intravascular ultrasound. PMID- 16308511 TI - Potent capillary isotachophoresis (cITP) for analyzing a marker of coronary heart disease risk and electronegative low-density lipoprotein (LDL) in small dense LDL fraction. AB - BACKGROUND: The potency and usefulness of capillary isotachophoresis (cITP) for assessing whole-serum lipoprotein profiles and quantifying electronegative low density lipoprotein (LDL) has been previously reported. METHODS AND RESULTS: A new cITP method to measure electronegative LDL in the small dense LDL fraction has been established. Both electronegative LDL and electronegative LDL in the small dense LDL fraction decreased after treatment with fenofibrate. CONCLUSIONS: This method appears to be useful for analyzing a marker of coronary heart disease risk and may be suitable for evaluating the effects of hypolipidemic agents. PMID- 16308512 TI - Medico-legal aspects of traumatic injury of the vertebrobasilar artery. AB - Based on our experiences with medico-legal autopsies, we analyzed traumatic injury of the vertebrobasilar artery (VBA) in traffic accident victims. VBA rupture occurred in six cases with traumatic medullary lesions, but in none with cervical cord injury. A relatively small external force applied to the head or neck can induce isolated traumatic subarachnoid hemorrhage (SAH), which accounts for approximately 11% of fatal head injuries. We examined the relationship between site of impact and site of vascular injury in 16 cases of isolated traumatic SAH. The results showed that the vertebral artery (VA) on the side of impact tended to be longitudinally injured in victims experiencing an external force equivalent to that of a fist punch to the head or face. There was no clear relationship between the site of impact and the site of vascular injury in victims who had sustained an external force greater than that of a fist punch. However, our results suggest that an external force causing rotation of the head was likely to cause tearing of the artery. In cases in which an external force equivalent to that of a fist punch was applied to the head, a raised blood alcohol level was a significant risk factor for artery rupture. Furthermore, a difference in the diameters of the left and right VAs was a risk factor for artery rupture. To determine why the VA, the most frequent site of arterial dissection, is also frequently the site of injury, we histologically examined normal and dissected VBAs. We also discuss medico-legal issues of the causal relationship between external force applied and rupture or dissection of the VA. PMID- 16308513 TI - High incidence of microbleeds in hemodialysis patients detected by T2*-weighted gradient-echo magnetic resonance imaging. AB - The incidence and characteristics of microbleeds in hemodialysis (HD) patients were investigated to elucidate the clinical significance with T(2)(*)-weighted gradient-echo magnetic resonance (MR) imaging. The 57 patients with chronic renal failure maintained by HD had no previous history of stroke. The control group consisted of 53 patients without previous history of stroke or chronic renal failure. The incidence and the number of microbleeds were assessed in the HD and control groups. The findings of microbleeds with T(2)(*)-weighted gradient-echo MR imaging were compared with those of T(1)- and T(2)-weighted MR imaging in HD patients. The incidence of microbleeds was significantly greater in the HD patients compared with the control patients. T(2)(*)-weighted gradient-echo imaging revealed a total of 44 microbleeds in 11 HD patients. T(2)-weighted imaging demonstrated 13 of 44 microbleeds as hyperintensity, whereas T(1) weighted imaging demonstrated 12 lesions as hypointensity. T(2)- and T(1) weighted imagings did not demonstrate any findings in 31 and 32 lesions, respectively. T(2)(*)-weighted gradient-echo MR imaging is effective to detect microbleeds which may be a predictor of intracerebral hemorrhage in HD patients and should be included in the protocol for the study of cerebrovascular disease, because T(2)- and T(1)-weighted MR imaging recognizes microbleeds as lacunar infarction. PMID- 16308514 TI - Evaluation of aneurysm stability after endovascular embolization with Guglielmi detachable coils: correlation between long-term stability and volume embolization ratio. AB - The correlation between volume embolization ratio (VER), the percentage of aneurysm volume occupied by coils, and aneurysm stability after endovascular treatment with platinum coils was studied in 86 patients who underwent embolization of 88 cerebral saccular aneurysms with standard platinum coils between March 1997 and January 2003. Radiographic follow up was possible of 62 aneurysms with greater than 70% obliteration on immediate post-procedure angiograms for more than 6 months. Immediate post-procedure evaluation found that 12 of 14 small (<4 mm), 12 of 38 medium (4-<10 mm), and two of 10 large (> or =10 mm) aneurysms had high (> or =25%) VER. In addition, the VER was high in 22 of 40 aneurysms with small (<4 mm) and four of 22 with wide necks (> or =4 mm). Evaluation of 57 aneurysms at 6 months after coil embolization found recanalization in two of 25 aneurysms with high VER and 21 of 32 with low (<25%) VER, indicating that aneurysms with high VER are significantly more stable than those with low VER (p < 0.01). VER strongly affects aneurysm stability, so high VER is desirable and a useful predictor of aneurysm stability. PMID- 16308515 TI - Simulation of endovascular neurointervention using silicone models: imaging and manipulation. AB - Silicone models of cerebral aneurysm and stroke pathology were produced by copying the images of clinical cases to evaluate the simulation of endovascular procedures. Conventional silicone models, silicone suture models made from separate aneurysm sac and parent artery sutured together, and animal vascular models made from vessels obtained from pigs were made. The models were incorporated in the training system for simulation of endovascular procedures under digital subtraction angiography control using a portable angiography system. The conventional silicone models provided very close reproduction of the aneurysm and vessels, even the fine branches. However, greater resistance or impact was felt, especially when passing the device through curved vessels. Stroke models were insufficiently dilated by percutaneous transluminal angioplasty or stenting. The silicone suture model was useful to avoid damage to models of giant aneurysm or models with small orifices during the lost wax procedure, whereas the projection of domes was somewhat inaccurate. The animal vascular model could not provide accurate aneurysm shape and vessel diameter, although the feel during manipulation was much more realistic. PMID- 16308516 TI - Surgical treatment of a lenticulostriate artery aneurysm. Case report. AB - A previously healthy 61-year-old man presented with basal ganglia hemorrhage caused by rupture of a small aneurysm arising from the distal lenticulostriate artery associated with moyamoya-like disease and manifesting as left hemiparesis and dysarthria. The patient underwent frontotemporal craniotomy. Neck clipping of the aneurysm was performed through the hematoma cavity. He was transferred for rehabilitation with left hemiparesis. Three-dimensional computed tomography angiography was very useful for preoperative planning and evaluation of the anatomical correlation between the aneurysm and the hemorrhage. PMID- 16308517 TI - Improvement of cerebral arterial stenosis associated with Basedow's disease. Case report. AB - A 29-year-old female presented with Basedow's disease manifesting as sudden vomiting, diarrhea, fever over 38 degrees C, transient aphasia, and numbness in her extremities. These symptoms were considered due to cerebral ischemia at a local clinic. Magnetic resonance angiography indicated stenosis of the bilateral distal internal carotid arteries and the bilateral proximal anterior cerebral and middle cerebral arteries. Thyroid swelling and exophthalmos were observed. She was transferred to our hospital. Endocrine function tests showed hyperthyroidism. The diagnosis was Basedow's disease. Her symptoms disappeared after receiving intravenous drip infusion of fluid replacement, and antithyroid and antiplatelet medication. After she became euthyroid, cerebral angiography and magnetic resonance angiography revealed improvement of the stenosis of the cerebral arteries. Stenosis of the terminal portion of the internal carotid artery associated with Basedow's disease is extremely rare. Conservative treatment mainly including antithyroid medications for Basedow's disease, and antiplatelet drugs and intravenous replacement fluid for the ischemic manifestations should be the first choice of treatment unless immediate vascular reconstruction is necessary. PMID- 16308518 TI - Traumatic cervical internal carotid artery occlusion in an infant. Case report. AB - A 10-month-old boy presented with traumatic internal carotid artery (ICA) occlusion caused by blunt injury after falling from a baby carrier attached to a standing bicycle. Physical examination found bruises on the scalp in the right temporal region and the right shoulder, but no wound in the neck. Chest radiography showed a right clavicular fracture. He developed left hemiparesis at 19 hours after the injury. Computed tomography revealed cerebral infarct and angiography showed cervical ICA occlusion. Conservative therapy with hemodilution was given under a diagnosis of cervical ICA occlusion caused by extension and rotational head injury. Traumatic cervical ICA occlusion due to blunt injury is not uncommon in adults, but extremely rare in infants. We would like to emphasize the dangers of leaving a baby unattended in a carrier attached to a standing bicycle. PMID- 16308519 TI - Central nervous system malignant lymphoma associated with Behcet's disease. Case report. AB - A 75-year-old woman presented with intracranial B-cell lymphoma associated with Behcet's disease manifesting as headache and dizziness. She had been treated with prednisolone for 17 years, and colchicine for 4 years under a diagnosis of incomplete Behcet's disease. Computed tomography revealed a 4-cm high density mass surrounded with diffuse edema in the right frontal lobe. Surgery removed a discolored red mass together with hematoma. The histological diagnosis was B-cell type malignant lymphoma. Malignant lymphoma is rarely associated with Behcet's disease, as only 10 cases have been reported. PMID- 16308520 TI - Anaplastic ganglioglioma of the cerebellopontine angle. Case report. AB - A 64-year-old woman presented with a rare anaplastic ganglioglioma in the right cerebellopontine angle manifesting as dizziness persisting for 2 weeks. Preoperative magnetic resonance (MR) imaging revealed a partially enhanced cystic lesion of the right cerebellopontine angle. The tumor was subtotally removed through a right lateral suboccipital craniectomy. The tumor was thought to originate from the brain stem with exophytic growth into the right cerebellopontine angle. Histological examination showed neoplastic ganglional and glial cells with anaplastic features such as mitosis, pleomorphism, and endothelial proliferation. The MIB-1 labeling index of the glial components was 40% to 60%. The diagnosis was anaplastic ganglioglioma (World Health Organization grade IV). She received postoperative radiotherapy but died of respiratory failure with tumor recurrence 11 months after the operation. Gangliogliomas usually have a good prognosis. Histological features of anaplasia and a high MIB 1 labeling index may be predictive of a poor clinical outcome. PMID- 16308521 TI - Subdural extension of recurrent olfactory neuroblastoma. Case report. AB - A 37-year-old man presented with olfactory neuroblastoma, which apparently recurred as diffuse extension in the subdural space of the cranial vault and spinal column 3 years after initial resection. Head and spinal magnetic resonance imaging with gadolinium demonstrated a subdural lesion. Cytological examination of the cerebrospinal fluid was negative. Histological examination of a biopsy specimen suggested recurrence of the olfactory neuroblastoma. This type of recurrence is very unusual. PMID- 16308522 TI - Intractable seizures associated with proximal migration of a ventriculoperitoneal shunt. Case report. AB - A 6-year-old girl, who had received a ventriculoperitoneal (VP) shunt using the Codman-Hakim programmable valve system at age 3 months, presented with intractable seizures. Neuroimaging studies showed migration of the proximal part of the system, including the prechamber, into the cranium through the right frontal burr hole. Electroencephalography showed spike-and-wave complexes in the right hemisphere including the site of the migration. The ictus was resolved following revision surgery. The clinical findings suggested the seizures were due to irritation of the brain parenchyma by the migrated system. Proximal migration of a VP shunt may cause both shunt failure and additional focal symptoms. PMID- 16308524 TI - [Gene therapy of virus replication with RNAi]. AB - RNA interference (RNAi) is the process of sequence-specific, post-transcriptional gene silencing initiated by double-stranded RNA (dsRNA). The short interference RNA (siRNA) cleaves target RNA even in mammalian cells without adverse effects of long dsRNA such as an interferon response, and works much more efficiently than antisense oligonucleotide and ribozyme. The clinical application of siRNA has been tried especially for the viral diseases. There are still important problems for application of gene therapy including off-target effect and gene delivery of siRNA, but a rapid progress can be expected because of the extremely high efficiency of siRNA. PMID- 16308525 TI - [Human herpesvirus latency and fatigue]. AB - Human herpesvirus (HHV)-6 and HHV-7 establish life-long latency, a hallmark of herpesviruses, reactivate frequently, and are shed in saliva. To investigate the viral reactivation, we have identified the latency-associated transcripts of HHV 6, and have revealed the partial mechanism of HHV-6 reactivation. HHV-6 established latency in the macrophage, kept a fairly stable intermediate stage between latency and reactivation, and the viral reactivation was induced by two or more factors. To identify the factor (s) of HHV-6 reactivation, we studied the association between HHV-6 reactivation and the work-induced fatigue in healthy adults. Reactivation of HHV-6 was examined for viral DNA by semi-quantitative PCR method. As a result, 88% of healthy adults shed the reactivated HHV-6 in the saliva during the fatigue, and 23% shed HHV-6 after holidays (approximately 1 week). The copy number of HHV-6 DNA was also reduced after holidays. In HHV-7, 52% of healthy adults shed the reactivated HHV-7 in the saliva during the fatigue, and 30% shed HHV-7 after holidays; however, there were no significant differences in their positive ratio and in the amount of viral DNA. These findings suggest that HHV-6 is reactivated during the work-induced fatigue, and HHV-6 reactivation can be an objective biomarker for fatigue. PMID- 16308526 TI - [Caveolar endocytosis and virus entry]. AB - The endocytic function of caveolae has been controversial for a long time. However, a real-time-imaging analysis of Simian virus 40 (SV40) 's entry in cells has indicated the existence of caveolar endocytosis during virus entry. The caveolae engulfed SV40 virions begin budding from plasma membrane depending on dynamin. SV40 enclosed in caveolae vesicles move to the caveosome, then to the endoplasmic reticulum. In addition, it was demonstrated that human coronavirus 229E enters the cell through caveolae. This review examines the involvement of caveolae in endocytosis used by the viral entry system. PMID- 16308527 TI - [Evolution of lentiviruses and receptor specificity]. AB - Lentiviruses consist of primate lentiviruses, ungulate lentiviruses and feline immunodeficiency virus (FIV). The primate lentiviruses utilize CD4 and chemokine receptors as a primary receptor and coreceptors, respectively. Recently we found that FIV utilizes CD134 and CXCR4 as a primary receptor and a coreceptor, respectively. FIV utilizes feline CD134 but not human CD134, whereas it can utilize both feline and human CXCR4. Exceptionally an FIV laboratory strain can infect human cells via CXCR4 only by the CD134-independent manner. Similarly several strains of primate lentiviruses also infect cells by the CD4-independent manner. In this review, the evolution of the lentiviruses and possible mechanism for lentiviral cross-species transmission is discussed. PMID- 16308528 TI - [Phylogenetic analysis and pathogenicity of tick-borne encephalitis virus from Japan and far-east Russia]. AB - Phylogenetic analysis of tick-borne encephalitis (TBE) virus revealed that Hokkaido strain of TBE virus evolved several hundreds years ago in far-east Russia. TBE virus strains in Irkutsk area were identified as Siberian subtype of TBE virus. BHK-cell adapted mutant of TBE virus showed lower neuro-invasive virulence in mice than parent virus. The mutant carried one amino acid substitution in envelope protein which resulted in increase of positive charge of the protein. The mutant-infected mice showed lower virus titers in bloods and spleens than the parent-infected mice. Infectious c-DNA clone of TBE virus Hokkaido strain was successfully generated and was applied to examine the neurovirulence in mice. One amino acid change in envelope protein and 2 amino acid changes in Ns5 protein showed a synergistic effect on reduced neurovirulence in mice. PMID- 16308529 TI - [Prion diseases as zoonosis]. AB - Prion diseases such as bovine spongiform encephalopathy (BSE) have been recognized as zoonosis since the existence of variant Creutzfeldt-Jakob disease (vCJD) was reported in 1996. BSE became a serious social problem even in Japan after the first BSE case was found in 2001. The incidence of BSE in EU and UK appears declining, and the vCJD incidence also shows a tendency to decrease. On the contrary, fears for the spread of BSE became actual problems: BSE occurrence outside of EU, transmission of vCJD by blood transfusion, and the first vCJD case in Japan. To prevent further spread and to reduce the risk of BSE, it is important to continue BSE screening/surveillance, removal of specified risk materials from food and feed chains, and effective feed regulation. For the disclosure and elimination of prion-contaminated blood, materials for medical and pharmaceutical products and so on, it is required to improve the sensitivity of prion detection methods. Furthermore, it is also important to establish therapeutics of human prion diseases. PMID- 16308530 TI - [Clue to the molecular mechanism of virulence of highly pathogenic H5N1 avian influenza viruses isolated in 2004]. AB - Highly pathogenic avian H5N1 influenza A viruses have spread throughout Asia since 2003. These viruses are highly lethal to birds and humans. Of the 74 confirmed human cases, 49 were fatal (as of Mar 30, 2005), raising concerns of a possible pandemic by these viruses. Despite the well-established pathogenicity of these viruses, the molecular mechanism for expressing such high virulence remains elusive. Thus, we examined the pathogenicity of the H5N1 viruses isolated in Vietnam in 2003-2004 using animal models (mouse, duck, and ferret). Viruses from humans were generally more pathogenic in mice and ferrets than those from birds. Indeed, one human isolate was even lethal to ferrets. The human isolate possessing Lys at amino acid position 627 of PB2 was more virulent than that possessing Glu at this position, underscoring the importance of Lys at this position 627 of PB2 for efficient growth in mammals. PMID- 16308531 TI - [West Nile fever/encephalitis as one of the arboviral infections]. AB - West Nile virus maintains natural infection cycle between birds and mosquitoes. It has been known that about 200 species of birds are infected with West Nile virus and the virus is isolated from more than 40 species of mosquitoes. This suggests that West Nile virus has an ability to be transmitted by many species of mosquitoes and infect many kinds of animals. Approximately 20% of infected humans develop symptoms. West Nile fever, an acute febrile illness, is the main disease, and meningitis and encephalitis (meningoencephalitis) occasionally occur. Cases with flaccid paralysis or polyneuritis have been recently reported. Thus, West Nile virus causes multiple types of symptoms in humans. The endemic area has expanded in North America and Siberia. West Nile virus may enter Japan in the near future; therefore, we should keep paying attention to the endemic and epidemic situations in the world. PMID- 16308532 TI - [Advances in antiviral chemotherapy]. AB - Establishment of selective antiviral chemotherapy has achieved dramatic improvement of the prognosis of several viral infections. It has been considered for a long time that, unlike bacterial infections, viral diseases cannot be successfully treated with chemotherapeutic agents, since viral replication mostly depends on the host-cellular machinery. In fact, some compounds were reported to inhibit viral replication even in the 1950s and 1960s, yet they were also quite toxic to the host cells. The first antiviral compound that strongly inhibits viral replication without affecting the uninfected cells is the anti-herpes agent acyclovir (ACV), which was discovered in the 1970s. Furthermore, in the 1980s, the world-wide epidemic of AIDS caused by human immunodeficiency virus type 1 (HIV-1) infection has dramatically accelerated the development of new antiviral agents. At present, most of the effective antivirals are targeted at virus specific enzymes, such as ACV for herpes virus thymidine kinase, zidovudine for HIV-1 reverse transcriptase, squinavir for HIV-1 protease, and oseltamivir for neuraminidase of influenza virus. These agents can be administered systemically without serious side effects. However, several drawbacks, including delayed toxicity and drug-resistance, are associated with long-term treatment with several antiviral agents mostly in highly active antiretroviral therapy for HIV-1 infection. Thus, it seems still mandatory to continue the search for more effective and less toxic compounds against various viral infections. PMID- 16308533 TI - [Respiratory syncytial virus]. AB - Human respiratory syncytial virus (RSV) is the most common worldwide cause of lower respiratory tract infections (LRI) in infants less than 6 months of age. The prophylaxis against RSV infection by vaccination has been unsuccessful because of its adverse effects. As antiviral drug, ribavirin spray (aerosol) had been used clinically and reduces the amount of virus load, without reducing the necessity of symptomatic therapy and the duration of hospitalization. Therefore RSV LRI has been treated mainly symptomatically. Recently humanized anti-RSV F protein monoclonal antibody was developed and prescribed for prevention in high risk infants such as premature ones and those with chronic lung and congenital heart diseases. It reduced the incidence of hospitalization significantly. It has been introduced in clinical use in Japan following to Western countries. On the other hand, a number of anti-RSV drugs have now been investigation; however, no valuable drugs for clinical use have been yet developed. PMID- 16308534 TI - [Progress in antiretroviral drugs]. AB - HIV-1, causative agent of acquired immunodeficiency syndrome, was identified in the early 1980s . The plague quickly spread throughout the world and today 40 million people are living with HIV/AIDS. The first anti-HIV drug "zidovudine", was discovered in 1985, and many other inhibitory compounds have been developed successfully in the last decade. Today, three classes 17 antiretroviral drugs are available in Japan. This article overviews the history of anti-HIV drug discovery, present HIV-1 treatment, and on-going drug discovery. PMID- 16308535 TI - [New antivirals for herpesviruses]. AB - The long-term treatment of herpesvirus infections with current antivirals leads to the development of drug-resistant viruses. Because currently available antivirals finally target the viral DNA polymerase, mutant resistant to one drug often shows cross-resistance to other drugs. This evidence highlights the need for the development of new antivirals that have the different viral targets. Recently, high-through-put screening of large compound collections for inhibiting specific viral enzymes, or in vitro cell culture assay, has identified several new antivirals. These include the inhibitors of helicase/primase complex, terminase complex, portal protein and UL97 protein kinase. This review will focus on these new compounds that directly inhibit viral replication. PMID- 16308536 TI - [Current approaches for developing new anti-HCV agents and analyses of HCV replication using anti-HCV agents]. AB - Currently, patients with hepatitis C virus (HCV) are mainly treated with interferon alone or in combination with ribavirin. However, because the virus is not eliminated from approximately one half of the patients by this treatment, alternative approaches to the treatment of HCV infection are needed. Recently, an HCV subgenomic replicon system has been established in which an HCV subgenomic replicon autonomously replicated in cultured cells. It enables us to screen for anti-HCV agents in cell culture system. Taking advantage of this system, we examined the effects of various types of compounds on the replication of HCV. Consequently, we found that a well-known immunosuppressant, cyclosporin A (CsA), had a strong suppressive activity on HCV replication, at least in cell culture system. This anti-HCV activity did not require the immunosuppressive feature of CsA. Through the investigation into the mechanism of anti-HCV effect of CsA, it was suggested that cyclophilin B, one of the cellular target molecules of CsA, played a significant role in HCV replication. Thus, searching for anti-HCV agents may lead to the elucidation of one of the mechanisms of HCV replication. PMID- 16308537 TI - [Anti-virals for influenza virus infection]. AB - Dramatic advances in the diagnosis and treatment of influenza in Japan has been made in recent years. Rapid diagnosis tests for influenza are routinely performed in Japanese hospitals. Both zanamivir and oseltamivir have been approved for the treatment of influenza since 2001, in addition to amantadine. Japan has the highest figure of neuraminidase inhibitor-use in the world because the treatment of influenza with neuraminidase inhibitors is covered by Japan's National Health Insurance program. Therefore, we should carefully observe the appearance of resistance strains and side effects to neuraminidase inhibitors. PMID- 16308538 TI - [Red sea bream iridoviral disease]. AB - The first outbreak of red sea bream iridoviral disease caused by red sea bream iridovirus (RSIV) was recorded in cultured red sea bream Pagrus major in Shikoku Island, Japan in 1990. Since 1991, the disease has caused mass mortalities of cultured marine fishes not only red sea bream but also many other species. The affected fish were lethargic and exhibited severe anemia, petechiae of the gills, and enlargement of the spleen. The causative agent was a large, icosahedral, cytoplasmic DNA virus classified as a member of the family Iridoviridae and was designated as red sea bream iridovirus (RSIV). The genome of RSIV is liner dsDNA and considered to be circularly permitted and terminally redundant like other iridoviruses. The length of physical map of RSIV genome is 112,415bp. An indirect immunofluorescence test with a monoclonal antibody and PCR are commonly used for the rapid diagnosis of RSIV infected fish in the field. For the control of this disease, a formalin-killed vaccine against red sea bream iridoviral disease was developed and now commercially available. PMID- 16308539 TI - [Molecular ecology of microalgal viruses]. AB - A great amount of virus particles exist in natural waters. Each virion is considered to have its own ecological role, affecting the maintenance and fluctuation of aquatic ecosystems. We have been studying viruses infectious to micro-plankton, especially those infecting phytoplankton. Red tides are caused by drastic increase in abundance of plankton. We succeeded in elucidating that viral infection is one of the most important factors determining the dynamics and termination of algal blooms by means of field survey and molecular experiments. In addition, we demonstrated that the interrelationship between viruses and their hosts are highly complicated, and might be determined by the molecular-structural difference of viral capsids among distinct virus ecotypes. Furthermore, in the process of our investigation on various aquatic algal viruses, their importance as genetic sources has also been suggested. In order to deeply understand the mechanism of aquatic ecosystem, more intensive studies as for aquatic viruses are urgently required. PMID- 16308540 TI - [Interaction mechanism of marine birnavirus (MABV) in fish cell lines]. AB - Marine birnavirus (MABV) is a member of the genus Aquabirnavirus of the family Birnaviridae. MABV is an unenveloped icosahedral virus about 60 nm in diameter with two genomes of double-stranded RNA. MABV adsorbed not only onto the cell surfaces of susceptible (CHSE-214 and RSBK-2) cells but also onto resistant (FHM and EPC) cells. Furthermore, the virus entered into the cytoplasm through the endocytotic pathway in CHSE-214, RSBK-2 and FHM cells but did not penetrate EPC cells. The virus was found to bind to an around 250 kDa protein on CHSE-214, RSBK 2, FHM and EPC cells. The syntheses of viral proteins pVP2, NS and VP3 and further proteolytic processing after viral infection were examined by using Western blot analysis. pVP2, NS and VP3 were detected in the cytosolic fractions of CHSE-214, RSBK-2 and FHM cells at 4 h after infection. At this time, VP3 underwent further proteolytic processing in the cytosolic fractions of CHSE-214 and RSBK-2 cells. The expression of pVP2, NS and VP3 increased and pVP2 and NS also underwent further proteolytic processing similar to VP3 in the cytosolic fractions of CHSE-214, RSBK-2 and FHM cells at 8 h after infection. The further proteolytic processing of VP3 was detected in the nuclear fractions of CHSE-214, RSBK-2, but VP3 was detected as a single band in the nuclear fraction of FHM cells. pVP2 and NS were detected as thin bands only in the nuclear fractions of CHSE-214 cells. The results of Western blot analysis demonstrated that pVP2, NS and VP3 are localized in the nuclear fraction when they were independently expressed in CHSE-214, RSBK-2, FHM and EPC cells. The expression pattern in the cytosolic fraction was identical among the four cell lines when pVP2 and NS were independently expressed. However, pVP2 and NS were not detected in the nuclear fraction of CHSE-214 cells. Further proteolytic processing of VP3 was detected in both cytosolic and nuclear fractions of RSBK-2 ,FHM and EPC cells (Low level in EPC cell), but not in CHSE-214 cells when VP3 was independently expressed. Then, the processes of preVP2 to form morphological assemblages in the presence of VP3 or the cleavage of VP3 into two proteins in CHSE-214 cells were studied. When preVP2- and VP3 were co-expressed, virion like particles (64 nm, diameter) were observed close to the nuclear membrane by electron microscopy. The co-expression of preVP2 and the cleaved VP3 proteins led to an efficient assembly of tubules (22 nm, diameter). Further important finds will be obtained by this infection system using 4 fish cell lines in the next couple of years. PMID- 16308541 TI - [Koi herpesvirus disease]. AB - Koi herpesvirus (KHV) disease emerged at the late 1990s, and has rapidly spread to the world. In Japan, KHV disease first occurred at October 2003. The disease resulted in mass mortality of wild carp as well as cultured carp. Until now, KHV infected carp were found in 42 out of 47 prefectures in Japan. Only carp Cyprinus carpio is susceptible to KHV, while goldfish, closely-related species to carp, is not. The affected carp swim lethargically. Sunken eyes and gill necrosis are frequently noticed, but no marked internal signs are observed. Optimal water temperature for the disease is 18-23 degrees C. Under 13 degrees C or over 28 degrees C, no death occurs. Keep at over 30 degrees C cures KHV disease, but can make the fish latent carriers. Because the fish do not get acquired immunity against KHV disease under low water temperature, the disease recurs with increase of water temperature. Isolation of KHV is difficult. KHV disease is diagnosed through epidemiological investigation, disease signs and PCR detection of KHV DNA. Vaccine development is ongoing for restart of culturing carp at KHV contaminated places. PMID- 16308542 TI - [Analysis on primate lentivirus genome dimerization in virion]. AB - The genomic RNA of retrovirus, including the primate lentivirus such as HIV, always form dimers in matured virions. It is likely that the presence of two genomes in one virion is advantageous for survival, providing an extra template that can be used when one RNA molecule is damaged, and/or giving genetic variety to their progeny. However, these ideas might not fully explain why the virion have to carry multiple identical RNAs in spite of the severe limitation of the space. We developed and utilized a novel system to investigate viral RNA dimerization in virion clearly and simply without affecting RNA packaging. The results of precise mapping of dimerization functional region strongly suggested that the RNA dimerization is one of the essential steps of RNA packaging. PMID- 16308543 TI - [WHO global action plan for laboratory containment of wild polioviruses (Second edition)]. PMID- 16308544 TI - Plant-based formulation for bronchial asthma: a controlled clinical trial to compare its efficacy with oral salbutamol and theophylline. AB - BACKGROUND: Plant-based medicine is the 3rd most popular choice of both adults (11%) and children (6%) suffering from Asthma. While several plant-based formulations have been reported for the treatment of asthma in the past, many authors have published their reservations on clinical trials carried out using complementary and alternative medicines. OBJECTIVES: The authors desired to eliminate the shortcomings of the earlier clinical trials carried out by many investigators in a structured study. Therefore, a 12-week randomized double-blind placebo-controlled clinical study was conducted to investigate the efficacy of a plant-based formulation (DCBT4567-Astha-15) in comparison with oral salbutamol, salbutamol + theophylline and a matching placebo in patients with reversible asthma. METHODS: Ninety-four patients between 15 and 50 years of age, showing 15% improvement in forced expiratory volume in 1 s (FEV(1)) 15 min after a bronchial challenge of inhaled salbutamol (200 microg) were recruited, and the end point of the study was determined as a 15% improvement in FEV(1) and clinical symptoms like dyspnoea, wheezing, cough, expectoration, disability, sleep disturbances and respiration rate. RESULTS: DCBT4567-Astha-15, salbutamol and salbutamol + theophylline patients showed statistically significant improvement in FEV(1), while placebo patients did not show any improvement. Fifty percent of DCBT4567 Astha-15, 48% of salbutamol, 58% of salbutamol + theophylline and 26% of placebo patients showed the desired 15% improvement in FEV(1). Improved mean FEV(1) values at the end of the trial indicated that the salbutamol - theophylline combination was superior followed by salbutamol and DCBT4567-Astha-15. Clinical symptoms like dyspnoea, wheezing, cough, expectoration, disability, and sleep disturbances were significantly reduced in DCBT4567-Astha-15 patients compared to patients of the other three arms. CONCLUSIONS: DCBT4567-Astha-15 was as efficacious as salbutamol (12 mg/day) or salbutamol (12 mg/day) in combination with theophylline (200 mg/day) in the treatment of reversible asthmatics. Quality of life of patients also improved with DCBT4567-Astha-15 drug treatment. PMID- 16308545 TI - Human airway smooth muscle cells express eotaxin in response to signaling following mast cell contact. AB - BACKGROUND: Asthma is a chronic inflammatory disease of the airways. Mast-cell (MC)-derived cytokines may mediate both airway inflammation and remodeling. It has also been shown that airway smooth muscle cells (ASMC) can be a source of proinflammatory cytokines. In the human airways, MC-ASMC cell interactions may have pivotal effects on modulating inflammation. OBJECTIVES: We wanted to know whether the production of eotaxin, an important proinflammatory cytokine, through a cell-to-cell contact mechanism of human ASMC activation by MC was mediated by p38 MAPK. METHODS: We cocultured normal humanASMC with a human MC line (HMC-1) and assayed for the production of eotaxin. RESULTS: When cultured together, human ASMC and HMC-1 contact induced eotaxin secretion. Separation of HMC-1 and human ASMC by a porous membrane inhibited this induction. Coculturing of human ASMC with HMC-1 induced increased expression of eotaxin gene mRNA. HMC-1-derived cellular membranes caused an increase in eotaxin production in human ASMC. Activation of p38 MAPK was also seen in cocultures by Western blot, whereas eotaxin production in cocultures was significantly inhibited by the p38 inhibitor SB203580. CONCLUSION: These novel studies reveal the importance of cell-to-cell interactions in the complex milieu of airway inflammation. PMID- 16308546 TI - Inhibitory effect of pravastatin on transforming growth factor beta1-inducible gene h3 expression in a rat model of chronic cyclosporine nephropathy. AB - BACKGROUND/AIMS: Overexpression of transforming growth factor beta1-inducible gene h3 (betaig-h3) is associated with renal scarring in several models of renal disease. We investigated the inhibitory effect of pravastatin on betaig-h3 expression in a rat model of chronic cyclosporin A (CsA)-induced nephropathy. METHODS: Adult Sprague Dawley rats kept on a low salt diet (0.05% sodium) were treated daily for 4 weeks with vehicle (olive oil, 1 ml/kg), CsA (15 mg/kg) or both CsA and pravastatin (20 mg/kg in drinking water). The effect of pravastatin on betaig-h3 expression was evaluated using in situ hybridization, immunohistochemistry, and immunoblotting. Functional parameters, histopathology (tubulointerstitial fibrosis, TIF, and arteriolopathy), and levels of transforming growth factor beta1 (TGF-beta1) and endothelial nitric oxide synthase were compared for the different treatment groups. RESULTS: Co administration of pravastatin significantly inhibited betaig-h3 mRNA production and gene expression within the tubulointerstitium of the CsA-treated kidneys, and this paralleled an attenuation of TIF (12.7 +/- 2.2 vs. 35.9 +/- 5.4%, p < 0.01 vs. CsA) and the expression of TGF-beta1 mRNA (279 +/- 40 vs. 719 +/- 85%, p < 0.01 vs. CsA). Pravastatin treatment reduced endothelial nitric oxide synthase protein levels and reversed the renal dysfunction caused by CsA. Neither CsA nor pravastatin affected total serum cholesterol or triglyceride levels in the treatment groups. CONCLUSION: Pravastatin thus effectively abrogated the upregulation of betaig-h3 gene expression and associated TGF-beta1 production, and this was associated with attenuated TIF in this model of chronic CsA-induced nephropathy. PMID- 16308548 TI - [Update for the clinician]. AB - Musculoskeletal ultrasound is of considerable value for the clinician, either if performed by himself or by the radiologist. If carried out with adequate equipments, it is an essential imaging modality and, in addition, has a definite role in clarifying the pathogenesis of different disorders and works well as a teaching tool. Advantages of musculoskeletal ultrasound are several and well known. Definite disadvantages include time dedicated to perform the examination, cost of top level equipments and difficulty to judge the examination quality by the clinician. Ultrasound is a useful diagnostic modality that can assess disease's activity, follow-up and recovery and can successfully guide diagnostic and therapeutic interventional procedures. PMID- 16308549 TI - [Tendons: inflammatory and degenerative pathology]. AB - Tendons may be affected by a variety of pathologic conditions, including those caused by overuse and inflammatory diseases. Sonography is a very sensitive means of detecting tendonous pathology because of its spatial resolution and its comparative and dynamic capabilities. Moreover its wide availability makes it the preferred first-line imaging modality in these cases. This paper reviews the inflammatory and degenerative conditions that may involve the tendons and their sonographic appearances. PMID- 16308550 TI - [Tendons: traumatic lesions]. AB - In the recent years there has been an increase in the number of tendon tears (TT) seen in routine daily outpatient practice secondary to the increasing popularity of sport activities. Tears require early diagnosis to ensure proper treatment and reduce functional impairment. Since local pain, edema and reflex muscle contraction can significantly limit the usefulness of clinical examination, imaging is usually required to confirm the clinical diagnosis, differentiate between partial and complete tear, and localize the retracted tendon stump. Several imaging modalities can be used in the evaluation of TT. Ultrasound is an efficient, dynamic, low cost and non invasive modality that is being increasingly utilized in the evaluation of the musculoskeletal system. It is well accepted by the acutely injured patient. The aim of this review article is to describe the ultrasound findings of the most common tendon tears. PMID- 16308551 TI - [Sonography of muscles]. AB - Sonography constitutes a method of choice in the evaluation of muscular structures. Traumatic lesions represent the majority of muscular injuries. A good anatomical knowledge of the weakness sites, as well as knowledge of traumatic signs and symptoms is essential in order to deliver a precise report to the referring physician. Sonography may suggest other causes of muscle lesions, tumoral or dystrophic, before confirmation by MRI. PMID- 16308552 TI - [Ultrasound of peripheral nerves]. PMID- 16308553 TI - [Value of Color Doppler of bones and joints]. PMID- 16308554 TI - [Superficial US of superficial bones]. AB - Although bones are not well imaged by US this imaging modality can be helpful in the assessment of bone surface and can be complementary to standard radiographs. A focal irregularity of the hyperechoic cortical line indicates a fracture, a cortical avulsion, a local bulging of the cortex or a foreign body related or not to previous surgery. Subperiosteal collections either purulent or hemorrhagic are easily detected and can be aspirated under US guidance if an infection is suspected. US also allows diagnosis of epiphyseal fractures when involving the distal epiphysis of the metatarsals, radial head, humeral head (Hill-Sachs fracture) growing cartilages... US examination of joints can detect osteophytes and marginal erosions (allowing early diagnosis of rheumatoid arthritis together with visualization of hyperaemic pannus and joint effusion) Cortical continuity in a location where in normal conditions a joint space is found indicates a synostosis. In children US, by directly visualizing the cartilaginous component of the non ossified bones, allows detection and serial follow-up of many congenital malformation (pes equinus...). US may be obtained in all patients where standard radiographs are not diagnostic because of it is efficient, non invasive and relatively inexpensive. PMID- 16308555 TI - [Dynamic ultrasonography]. AB - The ability to perform dynamic evaluation is a great advantage of ultrasound especially for musculoskeletal evaluation. Different manoeuvres are routinely used. The importance of the mobility of a structure or an articulation, but also the grade of compression of the lesion, can provide useful diagnostic information. For ligaments, the tension of each band is important and mobilisation is often able to depict some conflicts between the tendons and others structures. Muscle contraction is also an important element for making the diagnosis and, similar to a Valsalva manoeuvre for the diagnosis of a hernia. Interventional procedures are also facilitated by this dynamic evaluation. On the other hand, this great advantage requires the presence of a physician during the examination. PMID- 16308556 TI - [Interventional sonography]. AB - Diagnostic and therapeutic interventional procedures are rapidly expanding and, when guided by imaging, are more efficient then when performed with a blinded technique. Compared to fluoroscopy and CT, ultrasound does not utilize ionizing radiation. It can facilitate needle placement for arthrography, tenography or bursography or it can guide a variety of procedures such as aspiration, arthrocentesis, local steroid injections and needling of tendon calcifications. Technological improvements have increased the precision of ultrasound guidance and have contributed to reduce the risk of complication. Real time scanning allows simultaneous visualization of the target and of needle progression and has diminished the rate of complications, that are infrequent if the operator uses a strict sterile technique and respects the contraindications. PMID- 16308557 TI - [Paediatric musculoskeletal ultrasound]. AB - Ultrasound examination is often the most effective and sometimes the only practical means of examining children with soft tissue, tendon and joint diseases. This review will cover the important application of diagnostic ultrasound in children musculoskeletal disorders. We will consider topics where ultrasound is the only sensible means of examination such as developmental dysplasia of the hip, where it is the best choice of several possible methods such as painful hip and when it provides an alternative or adjunct to other imaging such as the examination of soft tissue masses. PMID- 16308558 TI - [Usefulness of musculoskeletal ultrasonography in a University Department of Radiology]. AB - The excellent architectural analysis of muscles, tendons and nerves, the dynamic assessment of theses structures and the easy comparison with the controlateral side explain the usefulness of ultrasonography in the assessment of many musculoskeletal diseases. This technique also represents a pleasant way to learn the musculoskeletal anatomy and allows a better understanding of the physiology and pathophysiology of the musculoskeletal system. Finally, musculoskeletal ultrasonography is more and more used in clinical and fundamental research studies. PMID- 16308559 TI - [Technical R & D and future applications]. AB - Medical imaging is closely related to technological progress. During the last twenty years considerable progress enabled sonography to assess musculoskeletal structures more and more accurately. Development of new high frequency transducers, harmonic imaging, Doppler sonography and contrast agents resulted in better image quality and increased diagnostic accuracy. Nevertheless, future technologies may provide new additional functional and qualitative possibilities. The limitations of musculoskeletal sonography are rather well established. Some are probably due to physical "obstacles" that future technology could resolve. Elastography and heterogeneous tissues focalisation are examples of experimental applications which could push back the limits of our current practice. But the future of sonography cannot be reduced to a purely technological dimension. It also depends on clinical and paraclinical evaluation as well as on widely available education programs directed to promote a wider diffusion and improved clinical use of sonography. PMID- 16308560 TI - Long-term follow-up study of health status in population living in methylmercury polluted area. AB - At present, it is important to follow-up on the health condition of inhabitants living in a methylmercury-polluted area surrounding Minamata City, paying attention to subclinical disorders not only of the central nervous system, which have been concentrically studied for more than 40 years, but also to other health issues as well. We have performed annual follow-up multiple health examinations on about 1,500 persons of ages more than 40 years old in Tsunagi Town near Minamata City each summer from 1984 to 2004. Case-control studies were designed to estimate the role of risk factors for various health issues using geographical differences to compare the verified patients. The results of the study are summarized as follows: (1) There were no significant differences in the prevalence of diseases associated with Minamata disease (MD) (e.g., liver diseases, renal diseases). (2) Subjective complaints, which were related not only to neurological but also to general complaints were consistently much more common in the polluted area than in the control area. (3) Five percent of the inhabitants who were not certified as MD patients had a high predicting index of MD. They could be affected by methylmercury poisoning. It is important to make a differential diagnosis. (4) No significant differences with respect to ADL by residential area were observed. However, MD patients who stayed in their homes showed some impairments of instrumental ADL. (5) It is important to take into consideration mental distress not only from the physical effects but also from the secondary social damage experienced through MD in these area. PMID- 16308561 TI - Arsenic-contaminated water and extent of acute childhood malnutrition (wasting) in rural Bangladesh. AB - To investigate whether children's nutritional status, especially the extent of wasting [acute malnutrition as measured by weight-for-height Z (WHZ) score] is associated with arsenic contamination of drinking water, a survey was carried out in two discrete rural areas located less than 30 km southeast and northeast of Dhaka, the capital of Bangladesh. 761 children between 7 and 14 years of age were randomly selected, one child per household. The arsenic concentration in the tube well water used by each child and the prevalence and intensity of infection of three geo-helminths [Ascaris lumbricoides, Trichuris trichiura and hookworm (Ancylostoma duodenale or Necator americanus)] were determined. Each child had their height and weight measured and WHZ score determined using the World Health Organization (WHO) reference values. Basic socio-demographic data and knowledge of arsenic poisoning were collected using a questionnaire. 62.2% of the households studied were using well water with arsenic concentrations above the 0.01 mg/L WHO guideline (i.e. arsenic-contaminated households). The mean WHZ score of all the 761 children was -0.843 (sd 1.227) but children living in contaminated households were significantly more wasted than children living in noncontaminated households (mean difference = -0.361, p 0.001). When the effects of the differences in socio-economic status and prevalence of geo-helminths were also taken into account, the difference in means remained highly significant ( 0.330, p = 0.006). This study suggests that arsenic contamination has a negative impact on children's acute nutritional status. PMID- 16308562 TI - Regulation of G0 entry by the Pho80-Pho85 cyclin-CDK complex. AB - Eukaryotic cell proliferation is controlled by growth factors and essential nutrients. In their absence, cells may enter into a quiescent state (G0). In Saccharomyces cerevisiae, the conserved protein kinase A (PKA) and rapamycin sensitive TOR (TORC1) pathways antagonize G0 entry in response to carbon and/or nitrogen availability primarily by inhibiting the PAS kinase Rim15 function. Here, we show that the phosphate-sensing Pho80-Pho85 cyclin-cyclin-dependent kinase (CDK) complex also participates in Rim15 inhibition through direct phosphorylation, thereby effectively sequestering Rim15 in the cytoplasm via its association with 14-3-3 proteins. Inactivation of either Pho80-Pho85 or TORC1 causes dephosphorylation of the 14-3-3-binding site in Rim15, thus enabling nuclear import of Rim15 and induction of the Rim15-controlled G0 program. Importantly, we also show that Pho80-Pho85 and TORC1 converge on a single amino acid in Rim15. Thus, Rim15 plays a key role in G0 entry through its ability to integrate signaling from the PKA, TORC1, and Pho80-Pho85 pathways. PMID- 16308563 TI - A cell-autonomous requirement for the cell cycle regulatory protein, Rb, in neuronal migration. AB - Precise cell cycle regulation is critical for nervous system development. To assess the role of the cell cycle regulator, retinoblastoma (Rb) protein, in forebrain development, we studied mice with telencephalon-specific Rb deletions. We examined the role of Rb in neuronal specification and migration of diverse neuronal populations. Although layer specification occurred at the appropriate time in Rb mutants, migration of early-born cortical neurons was perturbed. Consistent with defects in radial migration, neuronal cell death in Rb mutants specifically affected Cajal-Retzius neurons. In the ventral telencephalon, although calbindin- and Lhx6-expressing cortical neurons were generated at embryonic day 12.5, their tangential migration into the neocortex was dramatically and specifically reduced in the mutant marginal zone. Cell transplantation assays revealed that defects in tangential migration arose owing to a cell-autonomous loss of Rb in migrating interneurons and not because of a defective cortical environment. These results revealed a cell-autonomous role for Rb in regulating the tangential migration of cortical interneurons. Taken together, we reveal a novel requirement for the cell cycle protein, Rb, in the regulation of neuronal migration. PMID- 16308564 TI - Phosphorylation by casein kinase 2 induces PACS-1 binding of nephrocystin and targeting to cilia. AB - Mutations in proteins localized to cilia and basal bodies have been implicated in a growing number of human diseases. Access of these proteins to the ciliary compartment requires targeting to the base of the cilia. However, the mechanisms involved in transport of cilia proteins to this transitional zone are elusive. Here we show that nephrocystin, a ciliary protein mutated in the most prevalent form of cystic kidney disease in childhood, is expressed in respiratory epithelial cells and accumulates at the base of cilia, overlapping with markers of the basal body area and the transition zone. Nephrocystin interacts with the phosphofurin acidic cluster sorting protein (PACS)-1. Casein kinase 2 (CK2) mediated phosphorylation of three critical serine residues within a cluster of acidic amino acids in nephrocystin mediates PACS-1 binding, and is essential for colocalization of nephrocystin with PACS-1 at the base of cilia. Inhibition of CK2 activity abrogates this interaction and results in the loss of correct nephrocystin targeting. These data suggest that CK2-dependent transport processes represent a novel pathway of targeting proteins to the cilia. PMID- 16308565 TI - Beta-arrestin2 functions as a phosphorylation-regulated suppressor of UV-induced NF-kappaB activation. AB - NF-kappaB activation is an important mechanism of mammalian UV response to protect cells. UV-induced NF-kappaB activation depends on the casein kinase II (CK2) phosphorylation of IkappaBalpha at a cluster of C-terminal sites, but how it is regulated remains unclear. Here we demonstrate that beta-arrestin2 can function as an effective suppressor of UV-induced NF-kappaB activation through its direct interaction with IkappaBalpha. CK2 phosphorylation of beta-arrestin2 blocks its interaction with IkappaBalpha and abolishes its suppression of NF kappaB activation, indicating that the beta-arrestin2 phosphorylation is critical. Moreover, stimulation of beta2-adrenergic receptors, a representative of G-protein-coupled receptors in epidermal cells, promotes dephosphorylation of beta-arrestin2 and its suppression of NF-kappaB activation. Consequently, the beta-arrestin2 suppression leads to promotion of UV-induced cell death, which is also under regulation of beta-arrestin2 phosphorylation. Thus, beta-arrestin2 is identified as a phosphorylation-regulated suppressor of UV response and this may play a functional role in the response of epidermal cells to UV. PMID- 16308566 TI - A view of consecutive binding events from structures of tetrameric endonuclease SfiI bound to DNA. AB - Many reactions in cells proceed via the sequestration of two DNA molecules in a synaptic complex. SfiI is a member of a growing family of restriction enzymes that can bind and cleave two DNA sites simultaneously. We present here the structures of tetrameric SfiI in complex with cognate DNA. The structures reveal two different binding states of SfiI: one with both DNA-binding sites fully occupied and the other with fully and partially occupied sites. These two states provide details on how SfiI recognizes and cleaves its target DNA sites, and gives insight into sequential binding events. The SfiI recognition sequence (GGCCNNNN[downward arrow]NGGCC) is a subset of the recognition sequence of BglI (GCCNNNN[downward arrow]NGGC), and both enzymes cleave their target DNAs to leave 3-base 3' overhangs. We show that even though SfiI is a tetramer and BglI is a dimer, and there is little sequence similarity between the two enzymes, their modes of DNA recognition are unusually similar. PMID- 16308567 TI - 3-D structural and functional characterization of the purified KATP channel complex Kir6.2-SUR1. AB - ATP-sensitive potassium (K(ATP)) channels conduct potassium ions across cell membranes and thereby couple cellular energy metabolism to membrane electrical activity. Here, we report the heterologous expression and purification of a functionally active K(ATP) channel complex composed of pore-forming Kir6.2 and regulatory SUR1 subunits, and determination of its structure at 18 A resolution by single-particle electron microscopy. The purified channel shows ATP-ase activity similar to that of ATP-binding cassette proteins related to SUR1, and supports Rb(+) fluxes when reconstituted into liposomes. It has a compact structure, with four SUR1 subunits embracing a central Kir6.2 tetramer in both transmembrane and cytosolic domains. A cleft between adjacent SUR1s provides a route by which ATP may access its binding site on Kir6.2. The nucleotide-binding domains of adjacent SUR1 appear to interact, and form a large docking platform for cytosolic proteins. The structure, in combination with molecular modelling, suggests how SUR1 interacts with Kir6.2. PMID- 16308568 TI - P-TEFb is not an essential elongation factor for the intronless human U2 snRNA and histone H2b genes. AB - Phosphorylation of Ser2 of the heptapeptide repeat of the CTD of mammalian pol II by P-TEFb is associated with productive elongation of transcription of protein coding genes. Here, we show that the CTD of pol II transcribing the human U2 snRNA genes is phosphorylated on Ser2 in vivo and that both the CDK9 kinase and cyclin T components of P-TEFb are required for cotranscriptional recognition of the 3' box RNA 3' end processing signal. However, inhibitors of CDK9 do not affect transcription of the U2 genes, indicating that P-TEFb functions exclusively as an RNA processing factor in expression of these relatively short, intronless genes. We also show that inhibition of CDK9 does not adversely affect either transcription of an intron-less, replication-activated histone H2b gene or recognition of the histone gene-specific U7-dependent RNA 3' end formation signal. These results emphasize that the role of P-TEFb as an activator of transcription elongation can be separated from its role in RNA processing and that neither function is universally required for expression of mammalian pol II dependent genes. PMID- 16308569 TI - A promiscuous liaison between IL-15 receptor and Axl receptor tyrosine kinase in cell death control. AB - Discrimination between cytokine receptor and receptor tyrosine kinase (RTK) signaling pathways is a central paradigm in signal transduction research. Here, we report a 'promiscuous liaison' between both receptors that enables interleukin (IL)-15 to transactivate the signaling pathway of a tyrosine kinase. IL-15 protects murine L929 fibroblasts from tumor necrosis factor alpha (TNFalpha) induced cell death, but fails to rescue them upon targeted depletion of the RTK, Axl; however, Axl-overexpressing fibroblasts are TNFalpha-resistant. IL-15Ralpha and Axl colocalize on the cell membrane and co-immunoprecipitate even in the absence of IL-15, whereby the extracellular part of Axl proved to be essential for Axl/IL-15Ralpha interaction. Most strikingly, IL-15 treatment mimics stimulation by the Axl ligand, Gas6, resulting in a rapid tyrosine phosphorylation of both Axl and IL-15Ralpha, and activation of the phosphatidylinositol 3-kinase/Akt pathway. This is also seen in mouse embryonic fibroblasts from wild-type but not Axl-/- or IL-15Ralpha-/- mice. Thus, IL-15 induced protection from TNFalpha-mediated cell death involves a hitherto unknown IL-15 receptor complex, consisting of IL-15Ralpha and Axl RTK, and requires their reciprocal activation initiated by ligand-induced IL-15Ralpha. PMID- 16308570 TI - TLR agonists regulate PDGF-B production and cell proliferation through TGF beta/type I IFN crosstalk. AB - Transforming growth factor-beta (TGF-beta) and type I interferon (IFN) autocrine/paracrine loops are recognized as key mediators of signaling cascades that control a variety of cellular functions. Here, we describe a novel mechanism by which Toll-like receptor (TLR) agonists utilize these two autocrine/paracrine loops to differentially regulate the induction of PDGF-B, a growth factor implicated in a number of diseases ranging from tumor metastasis to glomerulonephritis. We demonstrate that CpG-specific induction of PDGF-B requires activation of Smads through TGFbeta1 autocrine/paracrine signaling. In contrast, polyinosinic:polycytidylic acid strongly represses CpG's as well as its own intrinsic ability to induce PDGF-B mRNA through type I IFN-mediated induction of Smad7, a negative regulator of Smad3/4. Furthermore, we have shown that this crosstalk mechanism translates into similar regulation of mesangial cell proliferation. Thus, our results demonstrate the importance of crosstalk between TGF-beta and type I IFNs in determining the specificity of TLR-mediated gene induction. PMID- 16308571 TI - CD4+ T cell-independent DNA vaccination against opportunistic infections. AB - Depletion or dysfunction of CD4+ T lymphocytes profoundly perturbs host defenses and impairs immunogenicity of vaccines. Here, we show that plasmid DNA vaccination with a cassette encoding antigen (OVA) and a second cassette encoding full-length CD40 ligand (CD40L), a molecule expressed on activated CD4+ T lymphocytes and critical for T cell helper function, can elicit significant titers of antigen-specific immunoglobulins in serum and Tc1 CD8+ T cell responses in CD4-deficient mice. To investigate whether this approach leads to CD4+ T cell independent vaccine protection against a prototypic AIDS-defining infection, Pneumocystis (PC) pneumonia, we used serum from mice vaccinated with PC-pulsed, CD40L-modified DCs to immunoprecipitate PC antigens. Kexin, a PC antigen identified by this approach, was used in a similar DNA vaccine strategy with or without CD40L. CD4-deficient mice receiving DNA vaccines encoding Kexin and CD40L showed significantly higher anti-PC IgG titers as well as opsonic killing of PC compared with those vaccinated with Kexin alone. Moreover, CD4-depleted, Kexin vaccinated mice showed a 3-log greater protection in a PC challenge model. Adoptive transfer of CD19+ cells or IgG to SCID mice conferred protection against PC challenge, indicating a role of humoral immunity in the protection. The results of these studies show promise for CD4-independent vaccination against HIV related or other opportunistic pathogens. PMID- 16308572 TI - Enhancement of vaccine-mediated antitumor immunity in cancer patients after depletion of regulatory T cells. AB - In this study, we investigated whether elimination of CD4+/CD25+ Tregs using the recombinant IL-2 diphtheria toxin conjugate DAB(389)IL-2 (also known as denileukin diftitox and ONTAK) is capable of enhancing the immunostimulatory efficacy of tumor RNA-transfected DC vaccines. We show that DAB(389)IL-2 is capable of selectively eliminating CD25-expressing Tregs from the PBMCs of cancer patients without inducing toxicity on other cellular subsets with intermediate or low expression of CD25. DAB(389)IL-2-mediated Treg depletion resulted in enhanced stimulation of proliferative and cytotoxic T cell responses in vitro but only when DAB(389)IL-2 was omitted during T cell priming. DAB(389)IL-2 significantly reduced the number of Tregs present in the peripheral blood of metastatic renal cell carcinoma (RCC) patients and abrogated Treg-mediated immunosuppressive activity in vivo. Moreover, DAB(389)IL-2-mediated elimination of Tregs followed by vaccination with RNA-transfected DCs significantly improved the stimulation of tumor-specific T cell responses in RCC patients when compared with vaccination alone. Our findings may have implications in the design of immune-based strategies that may incorporate the Treg depletion strategy to achieve potent antitumor immunity with therapeutic impact. PMID- 16308573 TI - Identification of human urinary trefoil factor 1 as a novel calcium oxalate crystal growth inhibitor. AB - Previous research on proteins that inhibit kidney stone formation has identified a relatively small number of well-characterized inhibitors. Identification of additional stone inhibitors would increase understanding of the pathogenesis and pathophysiology of nephrolithiasis. We have combined conventional biochemical methods with recent advances in mass spectrometry (MS) to identify a novel calcium oxalate (CaOx) crystal growth inhibitor in normal human urine. Anionic proteins were isolated by DEAE adsorption and separated by HiLoad 16/60 Superdex 75 gel filtration. A fraction with potent inhibitory activity against CaOx crystal growth was isolated and purified by anion exchange chromatography. The protein in 2 subfractions that retained inhibitory activity was identified by matrix-assisted laser desorption/ionization-time-of-flight MS and electrospray ionization-quadrupole-time-of-flight tandem MS as human trefoil factor 1 (TFF1). Western blot analysis confirmed the mass spectrometric protein identification. Functional studies of urinary TFF1 demonstrated that its inhibitory potency was similar to that of nephrocalcin. The inhibitory activity of urinary TFF1 was dose dependent and was inhibited by TFF1 antisera. Anti-C-terminal antibody was particularly effective, consistent with our proposed model in which the 4 C terminal glutamic residues of TFF1 interact with calcium ions to prevent CaOx crystal growth. Concentrations and relative amounts of TFF1 in the urine of patients with idiopathic CaOx kidney stone were significantly less (2.5-fold for the concentrations and 5- to 22-fold for the relative amounts) than those found in controls. These data indicate that TFF1 is a novel potent CaOx crystal growth inhibitor with a potential pathophysiological role in nephrolithiasis. PMID- 16308574 TI - Cross-reactive influenza virus-specific CD8+ T cells contribute to lymphoproliferation in Epstein-Barr virus-associated infectious mononucleosis. AB - The marked proliferation of activated CD8+ T cells is pathognomonic of EBV associated infectious mononucleosis (IM), common in young adults. Since the diversity and size of the memory CD8+ T cell population increase with age, we questioned whether IM was mediated by the reactivation of memory CD8+ T cells specific to previously encountered pathogens but cross-reactive with EBV. Of 8 HLA-A2+ IM patients, 5 had activated T cells specific to another common virus, as evidenced by a significantly higher number of peripheral blood influenza A virus M1(58-66)-specific T cells compared with healthy immune donors. Two patients with an augmented M1 response had tetramer-defined cross-reactive cells recognizing influenza M1 and EBV-BMLF1(280-288), which accounted for up to one-third of their BMLF1-specific population and likely contributed to a skewed M1-specific T cell receptor repertoire. These epitopes, with only 33% sequence similarity, mediated differential effects on the function of the cross-reactive T cells, which may contribute to alterations in disease outcome. EBV could potentially encode an extensive pool of T cell epitopes that activate other cross-reactive memory T cells. Our results support the concept that cross-reactive memory CD8+ T cells activated by EBV contribute to the characteristic lymphoproliferation of IM. PMID- 16308575 TI - Preferential migration of effector CD8+ T cells into the interstitium of the normal lung. AB - The respiratory tract is a primary site of infection and exposure to environmental antigens and an important site of memory T cell localization. We analyzed the migration and retention of naive and activated CD8+ T cells within the noninflamed lungs and quantitated the partitioning of adoptively transferred T cells between the pulmonary vascular and interstitial compartments. Activated but not naive T cells were retained within the lungs for a prolonged period. Effector CD8+ T cells preferentially egressed from the pulmonary vascular compartment into the noninflamed pulmonary interstitium. T cell retention within the lung vasculature was leukocyte function antigen-1 dependent, while the egress of effector T cells from the vascular to the interstitium functions through a pertussis toxin-sensitive (PTX-sensitive) mechanism driven in part by constitutive CC chemokine ligand 5 expression in the lungs. These results document a novel mechanism of adhesion receptor- and pulmonary chemokine dependent regulation of the migration of activated CD8+ T cells into an important nonlymphoid peripheral site (i.e., the normal/noninflamed lung). PMID- 16308576 TI - Are anxiety and depression addressed in primary care patients with chronic obstructive pulmonary disease? A chart review. AB - OBJECTIVE: Screening for mental illness in primary care is widely recommended, but little is known about the evaluation, treatment, and long-term management processes that follow screening. The aim of this study was to examine and describe the quality of mental health care for persons with chronic obstructive pulmonary disease (COPD) and anxiety/depressive disorders, as measured by adherence to practice guidelines. METHOD: This retrospective chart review examined data for 102 primary care and mental health care patients with COPD who were diagnosed, using Structured Clinical Interview for DSM-IV criteria, with major depressive disorder, dysthymia, depression not otherwise specified, generalized anxiety disorder, or anxiety not otherwise specified. Data were gathered from primary care progress notes from the year prior to enrollment in a randomized controlled trial (enrollment was from July 2002 to April 2004). We compared the care received by these patients over 1 year with that recommended by practice guidelines. Charts were abstracted using a checklist of recommended practice guidelines for diagnostic evaluation, acute treatment, and long-term management of anxiety and depressive disorders. RESULTS: Fifty (49%) of the 102 patients were recognized during the review year as having an anxiety or depressive disorder. Eighteen patients were newly assessed for depressive or anxiety disorders during the chart review year. Patients followed in primary care alone, compared with those who were comanaged by mental health care providers, were less likely to have guideline-adherent care. CONCLUSION: Depressive and anxiety disorders are recognized in about half of patients; however, guideline supported diagnostic evaluation, acute treatment (except for medications), and long-term management rarely occur in the primary care setting. To improve the treatment of depressive and anxiety disorders in primary care, the process of care delivery must be understood and changed. PMID- 16308577 TI - Risperidone treatment in 12 children with developmental disorders and attention deficit/hyperactivity disorder. AB - BACKGROUND: Risperidone is a novel antipsychotic drug that has been tried in the treatment of several child psychiatric disorders. In an open clinical study, we evaluated the safety and efficacy of risperidone in children with developmental disorder and behavioral problems including attention-deficit/hyperactivity disorder (ADHD). METHOD: Twelve patients aged 4 to 14 years who had a DSM-IV diagnosed developmental disorder and ADHD in addition to other behavioral problems, in particular aggression, were treated with risperidone for a period of up to 2 years with daily doses ranging from 1 to 3 mg. Data were gathered from December 2002 to December 2004. RESULTS: A positive clinical response was noted in 9 of the 12 patients within 3 months of study recruitment according to the Clinical Global Impressions-Improvement scale. Risperidone was well tolerated by all 12 patients. The most commonly reported side effect was sedation, which necessitated dosage reduction in 2 patients, but not discontinuation. CONCLUSIONS: Our findings suggest that risperidone may be an effective and safe treatment for children and adolescents with developmental disorder and disruptive behaviors. PMID- 16308578 TI - Major depressive disorder in hepatitis C: an open-label trial of escitalopram. AB - BACKGROUND: Depression is a common condition associated with hepatitis C and may be induced by interferon alfa, the primary treatment for hepatitis C. Depression is also a major barrier to the initiation of such treatment. This study examined the effect of escitalopram on measures of depression, quality of life, and tests of liver function in subjects with comorbid hepatitis C and depression. METHOD: Subjects with DSM-IV major depressive disorder and hepatitis C were included in this open-label study. The recruitment period was from October 2002 through February 2004. Treatment status with regard to interferon therapy was neither an inclusion nor an exclusion criterion. Subjects received escitalopram for 8 weeks starting at 10 mg/day. Dosage adjustments up to 20 mg/day were made after week 4, as deemed clinically necessary. Scores on the 17-item Hamilton Rating Scale for Depression (HAM-D-17) and the Clinical Global Impressions-Severity of Illness scale (CGI-S) and results of liver function tests (AST, ALT, GGT) were obtained at baseline, 2 weeks, 4 weeks, and 8 weeks. Medical Outcomes Study Short Form Health Survey (SF-36) ratings and Hopkins Symptom Checklist-90-Revised (SCL-90-R) scores were obtained at baseline and week 8. RESULTS: Eighteen subjects (12 female, 6 male) participated in this study. The mean daily dose of escitalopram at endpoint was 12.78 mg. Mean HAM-D-17 scores decreased significantly with treatment (t = 8.535, df = 17, p < .0001). Statistically significant improvement was also demonstrated on many subscales of the SF-36, the SCL-90-R, and the CGI S. Tests of liver function showed no significant changes. CONCLUSION: These results suggest that depression in patients with hepatitis C may be effectively and safely treated with escitalopram. PMID- 16308579 TI - Childhood bipolar disorder: a clinical vignette. AB - Bipolar disorder in children and adolescents can easily be misdiagnosed, as symptom patterns overlap with other mood disorders and attention deficit/hyperactivity disorder and commonly differ from adult presentations. A detailed description of the child's behavioral history and previous treatment response is critical to accurate diagnosis. Although studies with children are limited, a number of psychopharmaceuticals have been shown to provide therapeutic benefit. Parent/caregiver and educational support are essential components of successful treatment. This article describes an 11-year-old patient with bipolar disorder and then summarizes related treatment issues and options. PMID- 16308580 TI - Delirium-associated disulfiram and ethanol interactions. AB - BACKGROUND: Disulfiram, an agent used for the treatment of alcohol dependence, can exacerbate psychiatric syndromes (including psychosis, catatonia, delirium, depression, and mania) after extended use. However, delirium has yet to be reported following the short-term use of disulfiram in the setting of alcohol use. OBJECTIVES: We report a case with a neuropsychiatric presentation and discuss the prevention and the progression of delirium associated with an interaction of disulfiram and ethanol. CASE REPORT: We report the case of a 51 year-old woman who developed disorganized speech, diminished communication, a decrease in appetite, and thoughts of suicide 10 days after she began taking disulfiram (250 mg/day), to which she added 1 glass of alcoholic beverage for 2 days. Delirium developed in association with an interaction between disulfiram and alcohol. The patient met DSM-IV criteria for major depressive disorder, alcohol dependence, and delirium. DISCUSSION: Neuropsychiatric manifestations may develop in association with co-administration of disulfiram and alcohol; timely recognition and treatment are recommended. PMID- 16308582 TI - Treatment of irritable bowel syndrome: changing the paradigm. PMID- 16308581 TI - Psychotic symptoms in the elderly. AB - Lessons Learned at the Interface of Medicine and PsychiatryThe Psychiatric Consultation Service at Massachusetts General Hospital (MGH) sees medical and surgical inpatients with comorbid psychiatric symptoms and conditions. Such consultations require the integration of medical and psychiatric knowledge. During rounds, Dr. Stern and the chief resident discuss the diagnosis and management of conditions confronted. These discussions have given rise to rounds reports that will prove useful for clinicians practicing at the interface of medicine and psychiatry. PMID- 16308584 TI - About the association of medicine and psychiatry. PMID- 16308583 TI - Pleasant dreams. PMID- 16308585 TI - Carbidopa-levodopa-induced recrudescence of premorbid tic disorder in remission. PMID- 16308586 TI - Panic disorder, alcohol and substance abuse, and benzodiazepine prescription. PMID- 16308587 TI - Canadian Consensus on cardiac transplantation in pediatric and adult congenital heart disease patients 2004: executive summary. AB - Cardiac transplantation is an acceptable therapeutic option for the pediatric age group and for adult patients with congenital heart disease. There are a myriad of clinical diagnoses in these two patient populations. Survival has continued to improve, with graft half-lives of 14 years and greater in pediatric heart transplantation patients. There are issues unique to these patient populations in relation to heart transplantation for which the present document summarizes the relevant literature and presents management guidelines. Donor availability remains a major limiting factor in organ transplantation at present. Efforts need to be made to increase organ donor awareness, identify potential donors and aggressively manage marginal donors. Indications for transplantation and determination of timing of listing continue to be challenging due to a lack of evidence-based guidelines specifically for prognostic indices of outcome and pretransplant survival. The current status system for listing patients for transplantation does not necessarily reflect the typical clinical course of deterioration experienced by these two patient populations; therefore, consideration needs to be given to a parallel listing strategy. Evidence is accumulating pointing to an advantage to performing transplantations in patients in early infancy. ABO-incompatible heart transplantation has lead to a reduction in waiting time and waiting list mortality. Care of children after heart transplantation must take into consideration physical growth and multisystem development; stage of immunological maturation; intellectual, emotional and social maturation; educational activities; and other pediatric quality of life parameters. Post-transplantation issues are somewhat different, including rejection, coronary artery disease, malignancies and infections. Efforts need to be made to support multicentre trials to determine optimal treatment protocols. PMID- 16308588 TI - Treating the right patient at the right time: access to care in non-ST segment elevation acute coronary syndromes. AB - In 2004, the Canadian Cardiovascular Society formed an Access to Care Working Group with a mandate to use the best science and information available to establish reasonable triage categories and safe wait times for common cardiovascular services and procedures through a series of commentaries. The present commentary discusses the rationale for access benchmarks for urgent cardiac catheterization and revascularization, including hospital transfer in the setting of non-ST elevation acute coronary syndromes. The literature on standards of care, wait times, wait list management and clinical trials was reviewed. A survey of all cardiac catheterization directors in Canada was performed to develop an inventory of current practices in identifying and triaging patients. The Working Group recommended the following medically acceptable wait times for access to diagnostic catheterization and revascularization in patients presenting with acute coronary syndromes: for diagnostic catheterization and percutaneous coronary intervention, the target should be 24 h to 48 h for high-risk, three to five days for intermediate-risk and five to seven days for low-risk patients; for coronary artery bypass graft surgery, the target should be three to five days for high-risk, two to three weeks for intermediate-risk and six weeks for low-risk patients. All stakeholders must affirm the appropriateness of these standards and work continuously to achieve them. However, some questions remain around what are the best clinical risk markers to delineate the triage categories and the utility of clinical risk scores to assist clinicians in triaging patients for invasive therapies. PMID- 16308589 TI - Profile of the cardiovascular specialist physician workforce in Canada, 2004. AB - The Canadian Cardiovascular Society (CCS) examined the demand for cardiovascular care, the supply of specialist physicians to provide that care and physician survey data on human resource issues, including workload, wait times, satisfaction and future intentions. The CCS used data from the National Physician Survey 2004, the CCS Cardiovascular Specialist Physician Workforce Survey 2001, the Canadian Medical Association's Physician Resource Evaluation Template and procedural volumes from the Canadian Institute for Health Information to analyze key human resource planning issues for cardiologists and cardiac surgeons. There are indications in the 2004 survey data that the average workload continues to be very heavy, with an average workweek of between 55 h (cardiologists) and 64 h (cardiac surgeons), and an additional on-call responsibility of between 106 h (cardiologists) and 196 h (cardiac surgeons) per month, of which 38 h (cardiologists) to 45 h (cardiac surgeons) are spent with patients. As a result, many patients are waiting too long for nonurgent referrals. In addition to the toll that this heavy workload exacts from the current workforce and the long wait times for nonurgent referrals, the workload may also compromise the profession's ability to attract physicians into cardiovascular care. Medical graduates, particularly female graduates, tend to consider workload flexibility and predictability when choosing a specialty area. Supply projections already show a decline in the number of cardiovascular specialists per 100,000 elderly population over the next 15 years. If cardiology and cardiac surgery are perceived by female medical graduates (which now make up over 50% of all graduates) as unattractive areas of specialization, the profession may have difficulty attracting needed new entrants in the future. The CCS Workforce Project Team endorses and supports the 2003 Canadian Medical Association's recommendation for the federal government to establish an independent institute for health human resources to foster and conduct research on the supply, mix, distribution and sustainability of Canada's health workforce. The CCS should also renew its commitment to invest in related activities to improve access to cardiovascular care. The CCS Workforce Project Team also believes that the profession's ability to recruit and retain new medical graduates will depend on how well it addresses the identified quality of work life issues. PMID- 16308590 TI - The girls in the boys' club: reflections from Canadian women in cardiology. AB - With impending human resource needs in the field of cardiology, attention has turned to recruitment of female medical trainees. The authors explore the issues around balancing career and family. Possible solutions are considered, and all require a satisfactory work-life balance that will be able to attract young, talented physicians. PMID- 16308591 TI - Left ventricular assist device: can Canada afford this? AB - Heart transplantation remains a limited option in the treatment of patients with end-stage heart failure because of the shortage of donors and the high incidence of heart failure disease in the Canadian population. The current use of mechanical circulatory support systems is directed primarily at patients at imminent risk of death who are temporarily supported as a bridge to cardiac transplantation. Newer left ventricular assist devices, namely, the axial and centrifugal flow pumps, are smarter and more compact devices. Although used as bridge to transplantation, these devices are now seriously considered to become an option in the chronic treatment of patients with severe and unresponsive congestive heart failure. PMID- 16308592 TI - Impact of diabetes on 12-month outcomes following coronary artery bypass graft surgery: results from the ROSETTA-CABG Registry. AB - BACKGROUND: Diabetes mellitus is associated with poorer long-term outcomes following coronary artery bypass graft (CABG) surgery. However, little is known about the impact of diabetes mellitus on outcomes during the first 12 months following CABG. OBJECTIVES: To examine the relationship between diabetes mellitus and outcomes during the 12 months following CABG. METHODS: The Routine versus Selective Exercise Treadmill Testing after Coronary Artery Bypass Grafting (ROSETTA-CABG) Registry is a prospective, multicentre study examining the use of functional testing after CABG surgery. A total of 398 patients who were enrolled in the ROSETTA-CABG Registry were examined. Diabetic status was defined by medication use at discharge. Only patients undergoing a first successful CABG (all ischemic areas thought to be revascularized) were included. RESULTS: Among the 398 patients, 37 (9.3%) were receiving insulin, 67 (16.8%) were receiving oral hypoglycemic agents, and 294 (73.9%) were not receiving insulin or oral hypoglycemic agents. Insulin-treated patients had a higher 12-month incidence of composite clinical events consisting of readmission for unstable angina, myocardial infarction or death than did oral hypoglycemic-treated patients and nondiabetic patients (21.6% versus 4.5% and 6.0%, respectively; P=0.0003). Insulin-treated patients were also more likely to undergo repeat cardiac catheterization than were oral hypoglycemic-treated patients and nondiabetic patients (18.9% versus 8.8% and 7.9%, respectively; P=0.03). After controlling for other variables, use of insulin was independently associated with a composite of adverse clinical events (OR 3.80, 95% CI 1.5 to 9.6, P=0.005). CONCLUSIONS: During the 12-month period after a successful CABG, insulin-treated patients had a higher rate of adverse cardiac events than did patients receiving oral hypoglycemic agents and nondiabetic patients. These results suggest that diabetic patients may benefit from more aggressive surveillance during the first year after CABG surgery. PMID- 16308593 TI - The Canadian off-pump coronary artery bypass graft registry: a one-year prospective comparison with on-pump coronary artery bypass grafting. AB - BACKGROUND: The authors sought to examine in-hospital and one-year outcomes of off-pump coronary artery bypass grafting (CABG) and to determine the subgroups of patients most likely to benefit from the off-pump procedure in a regular surgical practice. METHODS: From March 2001 to December 2002, 1657 consecutive patients were treated with off-pump CABG and 1693 consecutive patients were treated with on-pump CABG. Propensity score modelling was performed to control for treatment and selection bias. A propensity-matched analysis was performed to identify factors associated with survival benefit from the off-pump procedure. RESULTS: The mortality was similar postoperatively and at one year after surgery. The rate of stroke was decreased in the off-pump group postoperatively (OR=0.49, 95% CI 0.23 to 1.06) and significantly at one year after surgery (OR=0.49, 95% CI 0.27 to 0.90). A significant reduction in acute renal dialysis and a significant increase in myocardial infarction rates were seen in off-pump patients during the initial hospitalization but these differences disappeared during the follow-up period. The number of grafts completed was significantly lower in off-pump CABG than in on-pump CABG (2.62+/-1.00 versus 3.36+/-0.92, respectively; P<0.001). Hospital length of stay and the percentage of patients who required mechanical ventilation were significantly lower in the off-pump group than in the on-pump group. At one year after surgery, the adjusted rate of coronary angiogram and revascularization was similar between the two groups, and the adjusted rate of self-reported angina and memory status was significantly better in the off-pump CABG group. Almost all subgroups of patients had a neutral effect or a survival benefit with the off-pump technique. CONCLUSIONS: The results from a Canada-wide multicentre registry showed the safety and effectiveness of off-pump CABG in most subgroups of patients in a regular surgical practice. PMID- 16308594 TI - [Coronary angioplasty in the acute phase of a myocardial infarction on a myocardial bridge]. AB - A myocardial bridge is usually asymptomatic but can cause myocardial ischemia, myocardial infarction or sudden death. Two occurrences of coronary angioplasty in the acute phase of an anterior myocardial infarction on a myocardial bridge are reported. The first case was first treated only with a balloon, and then with a stent 12 h later after a relapse of angina pectoris and the recurrence of a severe compression. The second case immediately benefited from a stent. A systematic control at six months has shown the absence of restenosis in the first case and an asymptomatic occlusion of the stent in the second case. Its deocclusion has revealed a myocardial bridge downstream of the stent. Myocardial stunning might have caused a decreased systolic compression by the bridge in the first case, and an underestimation of its actual length in the second case. Its regression is held responsible for these two relapses. A long active stent installed at high pressure could be used to treat myocardial bridges during myocardial infarctions. PMID- 16308595 TI - Statin therapy in Canadian patients with hypercholesterolemia: the Canadian Lipid Study -- Observational (CALIPSO). AB - BACKGROUND: Although statins are widely used to reduce low density lipoprotein cholesterol (LDL-C), there is little information about patient profiles, treatment patterns and goal achievement among statin-treated patients in Canada. OBJECTIVES: To assess the profile of statin-treated patients and to determine whether they are achieving recommended targets for LDL-C. METHODS: The Canadian Lipid Study -- Observational (CALIPSO) was a cross-sectional study involving Canadian physicians who were among the top statin prescribers. Each physician enrolled up to 15 patients who were at least 18 years of age with a diagnosis of hyper-cholesterolemia and who had been using a statin for at least eight weeks. Sociodemographics, coronary artery disease (CAD) risk factors, pretreatment and current lipid levels, and history of lipid-lowering therapy were reported for 3721 patients. RESULTS: Sixty-eight per cent of statin-treated patients were at high CAD risk according to the 2003 Canadian guidelines, 46.4% had established cardiovascular disease, 33.9% had diabetes and 59.5% had hypertension. Average LDL-C reductions of 32% (37% for high-risk patients) were initially required to reach goal. At the study visit, patients had been treated for an average of 4.3 years and 24.2% were using a high statin dose. Despite statin therapy, 27.2% of all patients and 36.4% of those at high CAD risk had not achieved LDL-C targets. For 67.4% of these patients, the current therapy was not modified at the study visit. CONCLUSIONS: Despite effective therapies, many treated patients are not achieving recommended LDL-C targets. Strategies should be implemented to promote achievement of lipid treatment goals for high-risk patients, thereby reducing the risk of cardiovascular events and their associated clinical and economic burdens. PMID- 16308596 TI - Coronary artery bypass graft surgery: do women cost more? AB - BACKGROUND: Coronary artery bypass graft surgery (CABG) in women has been associated with worse clinical outcomes than CABG in men. However, little is known about the impact of sex on the cost of CABG. OBJECTIVE: To examine the impact of sex on hospital course and the cost of CABG. METHODS: Hospital course and cost were examined among 2880 female and 9137 male patients from four Canadian and five American hospitals. Data were obtained from a resource and cost accounting system used by each of the nine hospitals. RESULTS: Among the 12,017 patients who underwent CABG, 24% (n=2880) were women and 76% (n=9137) were men. Women had a significantly longer length of stay (LOS) than did men (10.3+/-0.2 days and 8.9+/-0.08 days, respectively; P<0.0001) and a significantly higher in hospital mortality than did men (2.6% and 1.5%, respectively; P<0.0001). The total unadjusted cost was higher for women than for men both in Canada (US$11,200+/-268 and US$10,143+/-139, respectively; P<0.0001) and the United States (US$22,715+/-509 and US$19,906+/-269, respectively; P<0.0001). After adjusting for age and comorbid conditions, female sex was associated with a 10% increase in LOS (P<0.0001), a 97% increase in mortality (P=0.0006) and a 7% increase in overall cost (P<0.0001). CONCLUSION: Compared with men, women undergoing CABG had a modestly increased LOS and a higher mortality. Total in hospital cost was higher for women in each of the nine hospitals studied. Compared with other clinical variables, female sex is a relatively minor determinant of cost. Nevertheless, because of the expected increase in the number of women undergoing CABG in the future, this increased cost may translate into an important economic burden. PMID- 16308598 TI - Establishing waiting time targets and not meeting them. PMID- 16308599 TI - Tako-tsubo cardiomyopathy. AB - A 48-year-old woman presented to her local hospital with chest pain. Her electrocardiogram showed incomplete right bundle branch block, diffuse ST segment elevation and QTc prolongation. Shortly after admission, she became hypotensive and was referred for a coronary angiogram. Her coronary arteries were normal, but left ventriculography showed morphological changes typical of apical ballooning, as found in tako-tsubo cardiomyopathy. An echocardiogram confirmed complete akinesis of the left ventricular apex, with hyperkinesis of basal segments and an ejection fraction of 35% to 40%. Dynamic left ventricular outflow tract obstruction was also noted. After temporary support with intra-aortic balloon counterpulsation, fluids and noradrenaline, she recovered promptly. The present paper reviews the characteristic features of tako-tsubo cardiomyopathy. PMID- 16308597 TI - The evaluation of a formalized queue management system for coronary angiography waiting lists. AB - BACKGROUND: Lengthy waiting lists for coronary angiography have been described in many health care systems worldwide. The extent to which formal queue management systems may improve the prioritization and survival of patients in the angiography queue is unknown. OBJECTIVE: To prospectively evaluate the performance of a formal queue management system for patients awaiting coronary angiography in Ontario. METHODS: The coronary angiography urgency scale, a formal queue management system developed in 1993 using a modified Delphi panel, allocates recommended maximum waiting times (RMWTs) in accordance with clinical necessity. By using a provincial clinical registry, 35,617 consecutive patients referred into the coronary angiography queue between April 1, 2001, and March 31, 2002, were prospectively tracked. Cox proportional hazards models were used to examined mortality risk across urgency after adjusting for additional clinical and comorbid factors. RESULTS: Good agreement was determined in urgency ratings between scores from the coronary angiography urgency scale and implicit physician judgement, which was obtained independently at the time of the index referral (weighted kappa = 0.49). The overall mortality in the queue was 0.3% (0.47%, 0.26% and 0.13% for urgent, semiurgent and elective patients, respectively). Urgency, as specified by the coronary angiography urgency scale, was the strongest predictor of death in the queue (P<0.001). However, when patients were censored according to their RMWTs, mortality was similar across different levels of urgency. Consequently, up to 18.5 deaths per 10,000 patients could have potentially been averted had patients been triaged and undergone coronary angiography within the RMWT as specified by the coronary angiography urgency scale. CONCLUSIONS: The incorporation of the coronary angiography urgency scale as a formal queue management system may decrease mortality in the coronary angiography queue. The authors recommend its implementation in health care systems where patients experience excessive waiting time delays for coronary angiography. PMID- 16308600 TI - Cardiac rupture during stress echocardiography. AB - A case of acute cardiac rupture during dobutamine stress echocardiography testing that was performed on the sixth day after admission for an acute inferoposterior myocardial infarction is reported. Following successful surgical repair, the postoperative course was complicated by severe mitral regurgitation secondary to papillary muscle rupture. PMID- 16308601 TI - DDD pacemaker implantation in a patient with persistent left superior vena cava and absent right superior vena cava: a four-year follow-up report. AB - Persistent left superior vena cava coexisting with the absence of right superior vena cava is an uncommon anomaly, and sometimes requires permanent pacemaker implantation due to the relatively high incidence of conduction disturbances and arrhythmias. This anomaly makes the implantation of pacemaker leads more difficult; therefore, accurate preintervention diagnosis of this anomaly is valuable for the interventionalist. A patient in which the diagnosis of persistent left superior vena cava coexisting with absent right superior vena cava was made echocardiographically before permanent pacemaker implantation is presented. Acetylsalicylic acid was prescribed after pacemaker implantation to prevent a potentially fatal complication of coronary sinus thrombosis, and no complication occurred during the four-year follow-up. Some clues for the noninvasive diagnosis of this anomaly and techniques of pacemaker implantation are also described briefly. PMID- 16308602 TI - Diltiazem did not induce gingival overgrowth in rats. A clinical, histological and histometric analysis. AB - The administration of calcium channel blockers has been associated with gingival overgrowth. However, there are few studies in humans or animals that evaluated the effect of diltiazem on gingival tissues. The present study assessed the influence of diltiazem, at different dosages and treatment duration, on gingival tissues of rats, using clinical, histological and histometric analyses. Eighty young male rats were separated into eight groups according to the dosage and duration of treatment. Rats were treated for 20 or 40 days with a daily subcutaneous injection of 5, 20 or 50 mg/kg of body weight of diltiazem. The results confirmed that diltiazem did not induce gingival overgrowth in rats. For all animals, the evaluation did not show gingival alterations regardless of the dosages and periods of treatment. The histometric analysis showed no significant change in the area of epithelium and connective tissues, although after 40 days of treatment a decrease in the area of connective tissue was observed, without statistically significant difference from control groups. Within the limits of this study, we suggest that diltiazem did not induce gingival overgrowth. PMID- 16308603 TI - Enamel matrix proteins associated with GTR and bioactive glass in the treatment of class III furcation in dogs. AB - This study investigated, both histologically and histometrically, the efficacy of enamel matrix derived proteins (EMD) associated with bioactive glass (BG) and an absorbable membrane in the treatment of class III furcation defects in mongrel dogs. After surgical defect creation and chronification, the lesions were randomly divided into three groups according to the treatment employed: Test Group 1--EMD + BG + membrane, Test Group 2--EMD + membrane and Control Group--BG + membrane. After a 90-day healing period, the dogs were sacrificed. The descriptive analysis and the histometric data showed similar results for the experimental groups in all studied parameters (MANOVA, p > 0.05). The association of Emdogain with bioglass and GTR, or with GTR only, showed similar results when compared with the ones obtained with bioglass associated with membrane in the treatment of class III furcation defects in dogs. The three modalities of treatment showed partial filling of the furcations, with bone and cementum regeneration limited to the apical portion of the defects. PMID- 16308604 TI - Increase of condylar displacement between centric relation and maximal habitual intercuspation after occlusal splint therapy. AB - The present study assessed condylar displacement between initial maximal habitual intercuspation (MHI) and centric relation (CR), recorded after using a deprogramming occlusal splint for an average period of 7.8 +/- 2.1 months prior to any orthodontic treatment. The sample consisted of 22 subjects, 11 male and 11 female, with an average age of 14.2 +/- 1.4 years, with Class II malocclusion and with no apparent signs or symptoms of temporomandibular dysfunction (TMD). Condylar displacement was measured using a Panadent axis position indicator in decimal fractions of a millimeter. The original mean vertical displacements and the corresponding standard deviations were 4.24 +/- 2.53 mm and 3.86 +/- 2.72 mm, respectively, for the right and left sides. Because a significant negative correlation was observed between original condylar displacements and age factors, the displacement values were statistically adjusted to 2.74 +/- 2.00 mm and 2.44 +/- 1.93 mm. On the horizontal plane, the mean displacements measured were -0.72 +/- 1.53 mm on the right side and -0.51 +/- 1.98 mm on the left. The mean displacement on the transversal plane was 0.03 +/- 0.87 mm. A comparison between these values and those observed in non-deprogrammed groups, as well as those published in the related literature, indicates that use of occlusal splints results in greater mean condylar displacement values, especially vertically, between CR and MHI positions, which contributed to a more accurate orthodontic diagnosis. PMID- 16308605 TI - In vitro biocompatibility tests of two commercial types of mineral trioxide aggregate. AB - Recently, regular and white mineral trioxide aggregate (MTA) are being used in Dentistry as retrofilling materials. Genotoxicity and cytotoxicity tests form an important part of cancer research and risk assessment of potential carcinogens. Thus, the goal of this study was to examine the genotoxicity and cytotoxicity of regular and white MTA in vitro by the single cell gel (comet) assay and trypan blue exclusion test, respectively. Mouse lymphoma cells were exposed to two presentation forms of MTA at final concentrations ranging from 1 to 1,000 microg/mL for 3 h at 37 degrees C. The results showed that both compounds tested did not produce genotoxic effects at all concentrations evaluated. Likewise, no statistically significant differences (p > 0.05) were observed in cytotoxicity. Taken together, our results suggest that regular and white MTA are not genotoxins and are not able to interfere in cellular viability as assessed by single cell gel (comet) assay and trypan blue assay, respectively. PMID- 16308606 TI - Demineralization effect of EDTA, EGTA, CDTA and citric acid on root dentin: a comparative study. AB - The purpose of this study was to biochemically compare the decalcifying effects of 1% EDTA (pH 7.4), 1% EGTA (pH 7.4), 1% CDTA (pH 7.4), 1% citric acid solutions (pH 1.0 and 7.4) and saline solution (control) on root dentin. Forty-eight single rooted teeth were used in this study. The canals were instrumented by the step back technique and the roots were randomly divided into six equal experimental groups (n = 8) according to the irrigating agent tested. A total of 30 microL of each solution was pipetted into the root canal and allowed to set undisturbed for 5 minutes. After this time, 15 microL of the solutions were removed from each canal using a Hamilton syringe and placed in a container with 5 mL of deionised water. The microg/mL concentration of calcium ion (Ca2+) extracted from the root canal samples was determined using inductively coupled plasma-atomic emission spectrometry (ICP-AES). Data were analysed by means of the Kruskal-Wallis and Mood's median tests. Citric acid solution at pH 1.0 removed more calcium than at pH 7.4 and than the other chelating solutions tested (p < 0.05). No differences were observed between EDTA and EGTA. Both EDTA and EGTA removed significantly more calcium than CDTA and citric acid at pH 7.4 (p < 0.05). There were no differences between citric acid at pH 7.4 and saline solution, which had the least efficacy for Ca2+ extraction (p > 0.05). These results indicate that citric acid at pH 1.0 is a good alternative as an irrigating solution to remove the smear layer and facilitate the biomechanical procedures. PMID- 16308607 TI - Brazilian gutta-percha points. Part I: chemical composition and X-ray diffraction analysis. AB - Eight nonstandardized gutta-percha points commercially available in Brazil (Konne, Tanari, Endopoint, Odous, Dentsply 0.04, Dentsply 0.06, Dentsply TP and Dentsply FM) were analysed chemically and by X-ray diffraction, and their chemical compositions were compared. The organic fraction (gutta-percha polymer and wax/resin) of the gutta-percha points was separated from the inorganic fraction (ZnO and BaSO4) by dissolving them in chloroform. The gutta-percha polymer was precipitated with acetone. The inorganic fraction was analysed by elemental microanalysis. Energy-dispersive X-ray microanalysis (EDX) and X-ray diffraction were employed to identify the chemical elements and compounds (barium sulfate and zinc oxide). The barium sulfate content was calculated based on the percentage of sulfur found in the elemental microanalysis. All analyses were repeated three times. The means and standard deviations of the percentage by weight of gutta-percha in the points were: Konne (17.6 +/- 0.30), Tanari (15.2 +/ 0.30), Endopoint (16.7 +/- 0.23), Odous (18.8 +/- 0.20), Dentsply 0.04 (15.7 +/- 0.17), Dentsply 0.06 (16.6 +/- 0.17), Dentsply TP (21.6 +/- 0.15) and Dentsply FM (16.3 +/- 0.23). The means and standard deviations of the zinc oxide content were: Konne (79.9 +/- 0.10), Tanari (81.9 +/- 0.07), Endopoint (81.3 +/- 0.40), Odous (79.7 +/- 0.26), Dentsply 0.04 (77.9 +/- 0.03), Dentsply 0.06 (78.2 +/- 0.07), Dentsply TP (69.8 +/- 0.19) and Dentsply FM (72.6 +/- 0.70). The method utilized was appropriate to quantify gutta-percha, wax/resin, zinc oxide and barium sulfate. Cone brands without barium sulfate were found. An unusual high wax/resin percentage was detected in Dentsply FM (p = 0.0003). Dentsply TP showed the highest gutta-percha percentage. PMID- 16308608 TI - Application of the total etching technique or self-etching primers on primary teeth after air abrasion. AB - Since the use of air abrasion has grown in pediatric dentistry, the aim of this study was to evaluate, by means of shear bond strength testing, the need to use the total etching technique or self-etching primers on dentin of primary teeth after air abrasion. Twenty-five exfoliated primary molars had their occlusal dentin exposed by trimming and polishing. Specimens were treated by: Air abrasion + Scotchbond MultiPurpose adhesive (G1); 37% phosphoric acid + Scotchbond MP adhesive (G2); Clearfil SE (G3); Air abrasion + 37% phosphoric acid + Scotchbond MP adhesive (G4); Air abrasion + Clearfil SE (G5). On the treated surface, a cylinder of 2 mm by 6 mm was made using a composite resin (Z100). Duncan's test showed that: (G2 = G3 = G5) > (G1 = G4). The use of a self-etching primer on air abraded dentin is recommended to obtain higher bond strengths. PMID- 16308609 TI - Response of osteoblastic cells to titanium submitted to three different surface treatments. AB - In the complex process of bone formation at the implant-tissue interface, surface properties are relevant factors modulating osteoblastic function. In this study, commercially pure titanium (cp Ti) samples were prepared with different surface characteristics using chemical attack with a sulfuric acid/hydrochloric acid based solution (treatment A); chemical attack plus anodic oxidation using phosphoric acid (treatment B); and chemical attack plus thermal oxidation followed by immersion in a sodium fluoride solution (treatment C). The samples were characterized by scanning electron microscopy (SEM), contact profilometry and contact angle. The biological performance of the prepared surfaces was evaluated using mice osteoblastic cell cultures for up to 21 days. Cells seeded on the different titanium samples showed similar behavior during cell attachment and spreading. However, cellular proliferation and differentiation were higher for samples submitted to treatments A and C (p < or = 0.05; n = 3), which were less rough and showed surface free energy with smaller polar components. PMID- 16308610 TI - Comparison among dental, skeletal and chronological development in HIV-positive children: a radiographic study. AB - The goal of this study was to evaluate skeletal, dental and chronological development in an HIV-positive group of children, as compared with a control group, during a four-year period. Panoramic radiographs and hand and wrist radiographs of 60 children were taken. The children, of both sexes, aged 5 years and 2 months to 15 years and 5 months, were selected as follows: 30 HIV-positive volunteers who had acquired the disease vertically, and 30 volunteers who did not present the HIV infection or any other systemic disease. All radiographs were technically standardized and analyzed according to criteria established by Nolla (dental age), Greulich and Pyle (bone age), and Eklof and Ringertz (bone age). The results were submitted to Student's t-test at a 5% level of significance. Based on the comparison between the chronological age and the dental or the skeletal age, significant differences were observed between HIV-positive and HIV negative children, both in 1999 and in 2003 (p < or = 0.05). Considering the results obtained with the methodology used, it was concluded that HIV-positive children of both sexes presented delayed bone development despite the administration of antiretroviral drugs, and that HIV-positive female children presented younger dental ages compared with their chronological ages in 1999 and in 2003; and HIV-positive males, in 1999. PMID- 16308611 TI - DNA extraction from human saliva deposited on skin and its use in forensic identification procedures. AB - Saliva is usually deposited in bite marks found in many homicides, assault and other criminal cases. In the present study, saliva obtained from volunteers was deposited on skin and recovered for DNA extraction and typing in order to evaluate its usefulness for practical case investigation and discuss the contribution of forensic dentistry to saliva DNA typing. Twenty saliva samples were collected from different donors and used as suspects' samples. Five of these samples were randomly selected and deposited (250 microl) on arm skin. Saliva was collected from skin using the double swab technique. DNA from saliva and skin deposited saliva samples was extracted by the phenol-chloroform method. DNA samples were amplified by PCR for DNA typing using a set of 15 STRs. The recovery of DNA from saliva deposited in the skin was 14 to 10 times lower than DNA quantity from saliva samples. DNA typing was demonstrated in 4 of 5 deposited saliva samples, the likelihood ratios estimated for these samples based on data of the Brazilian population were 1:11, 1:500, 1:159.140 and 1:153.700.123. Our results indicate that standardized procedures used for DNA collection and extraction from skin-deposited saliva can be used as a method to recover salivary DNA in criminal cases. However, it is important to observe that DNA recovery in forensic samples can be difficult. This study suggests that the analysis of saliva deposited on skin be incorporated into a criminal investigation since it may have great discriminatory power. PMID- 16308612 TI - Assessment of methyl methacrylate vapor toxicity on the rat tracheal epithelium. AB - Methyl methacrylate (MMA) is a monomer that is polymerized into resin by light and heat, producing a clear, resistant, durable and relatively inert plastic material. Because of these characteristics, MMA is largely used in Medicine as bone cement and in Dentistry, in dental braces and prostheses, thus generating continuous interest in its toxicity. Experimental and clinical studies have documented that monomers may cause a wide range of adverse health effects. The most important occupational exposure route of MMA is by inhalation. This study aims to evaluate the toxicity of MMA to the tracheal epithelium, according to the time of exposure. For this purpose, two experimental groups of rats were exposed to MMA by inhalation under poor ventilation: one group (n = 36) was exposed permanently, and the other (n = 36) was exposed during 8 hours per day, without water and food supply during the exposure period. A control group (n = 8) received normal air supply. Twelve animals of each study group were sacrificed after 5, 8 and 10 days of exposure together with two or four control animals. Twenty-nine (80.5%) of the rats continuously exposed to MMA developed inflammation on the tracheal epithelium, as well as 58.33% (n = 21) of those exposed 8 h/day and 87.5% (n = 7) of the control rats. No association was observed between the inflammatory process and MMA exposure; no significant alterations in the tracheal epithelium thickness were observed. Further studies on longer exposure times and analysis of other parameters will have to be conducted to exclude the possibility of tracheal damage by vapors of MMA. PMID- 16308613 TI - Viral coinfection in the oral cavity of HIV-infected children: relation among HIV viral load, CD4+ T lymphocyte count and detection of EBV, CMV and HSV. AB - Viral coinfection in the oral cavity associated to HIV infection was evaluated in 180 children from birth to 13 years of age of both sexes. The oral examinations were performed at the Pediatric AIDS Outpatient Clinic, Sao Lucas Hospital and Clinic Hospital, both in Porto Alegre, Brazil and at the School of Dental Medicine, University Hospital Center, State University of New York at Stony Brook, USA. The aim of this study was to identify the presence of viral infections in the oral cavity. PCR technique was used to determine opportunistic viral infections caused by CMV, EBV, and HSV in mucosal swabs. A high frequency of viral infection was detected in the oral cavity of HIV-infected children determined by the PCR technique. HIV-infected children with viruses had a favorable CD4+ T lymphocyte count and unfavorable viral load. PMID- 16308614 TI - Brazilian nursing in the age of information and knowledge. PMID- 16308615 TI - The global network of WHO Collaborating Centres for nursing and midwifery development: a policy approach to health for all through nursing and midwifery excellence. AB - In response to the complexities of health services delivery within the context of cultures, world events, medical technologies, and natural and manmade disasters, WHO recognized the need for external resources to assist it to meet its goals. In line with WHO's policy that research in the field of health is best advanced by assisting, coordinating and utilizing the activities of existing institutions, WHO Collaborating Centres were developed to support WHO with external expertise at the local, country and international levels. Of the 1,000 Collaborating Centres worldwide representing all the major health disciplines, 36, in 19 different countries, are dedicated to nursing and midwifery services, education, research and policy. These 36 Centres have joined forces as the Global Network of WHO Collaborating Centres for Nursing and Midwifery Development. Since its inception in 1987, the Global Network has developed into a significant force in international nursing leadership. Its far-reaching sphere of influence, its communications technology and its ability to develop collaborative projects, enable it to have a significant impact on the health of the people of the world. PMID- 16308616 TI - Follow-up of the child's motor abilities in day-care centers and pre-schools. AB - The aim was to describe the results of a developmental screening test in a cohort of 30 children attending 3 day-care centers in the city of Sao Paulo and develop hypotheses about these results. Thirty children were evaluated three times, during two years, by using the DDST. In the gross motor area the results improved (Tests of the Signs, 1st to the 3rd evaluation, p= 0.038*). There was a relationship between the age of the child (2 to 3 years) and worst results (Fisher's Exact test, p= 0.013*). In the fine motor area the results improved (Tests of the Signs, 1st to the 2nd evaluation, p= 0.031*). So, the development of motor abilities improved after the children started attending the day-care centers. We must undertake more researches in order to attribute the specific influences of each area: day-care center and/or family. PMID- 16308617 TI - "Without money you're nothing": poverty and health in Mexico from women's perspective. AB - The objective of this qualitative study was to get to know poor Mexican women's experience of poverty in relation to health care. Forty-nine interviews were carried out with poor adult women in Mexico (between 35 and 65 years old). Three central elements were detected in relation to the women's experience of poverty and health care: their socio-economic dependence on their family; the notion of social belonging in their experience with health care rights, reflected in the idea and acceptance that, due to their poverty, they can only be attended at philanthropic institutions; and the existence of survival mechanisms when facing an illness. In recovering the experience of poor women in relation to their health care, we identified that there is a clear idea that, if women had had economic resources, their health problem would have been solved differently. They are also convinced that, due to being poor, they have to content themselves with bad-quality medical care. This conformity finally makes them resign to the fact of either loosing a part of their own body, or even just waiting for death. PMID- 16308618 TI - [Adhesion to anti-retroviral therapy by individuals with HIV/AIDS seen at an institution in the interior of Sao Paulo]. AB - Inadequate adherence to highly active anti-retroviral therapy (HAART) provokes important secondary effects in people living with aids. The objective was to identify the factors that make HAART adherence easy or difficult, according to aids patients attended at a university hospital in the state of Sao Paulo, Brazil. We interviewed 200 diagnosed aids patients using HAART for at least 6 months. Patients were interviewed individually, using a semi-structured design. Qualitative and quantitative analysis was used. 59% of the participants were men; average age was 38.2 years; 51% did not finish basic education; 50.5% did not perform any remunerated work. Patients had been using anti-retroviral agents for an average time of 5 years. The number of anti-retroviral pills ranged from 3 to 24. The main difficulties mentioned for adherence were: taste, size, number, smell of pills (40.0%); intense collateral effects (14.4%); psychological factors (13.7%); different times to take the pills (10.8%). Patients mentioned the following facilitators: coincidence of times to take the drugs (26.2%), no facility (16.4%), and administration associated to some habit (16.0%). The nursing group needs to reinforce supervised surveillance, educational and intervention actions. PMID- 16308619 TI - [Time of permanence of nurses at a school-hospital and expenses related to admission, resignation and hiring of a new professional]. AB - This descriptive study aimed to find out how long nurses stay at a hospital institution and the relationship with expenses in terms of hours/salary regarding admission, resignation and hiring of a new nurse. We analyzed the resignation records of 109 nurses (2000-2002) from a hospital in the eastern region of Sao Paulo, provided by the Human Resource Department in 2003, as well as the amounts spent on hours/salary of the personnel involved in the selection process, admission, training and resignation. The study showed that the nurses stayed for a short period (average of 20 months and median 18). 30.53% of the nurses left the hospital in less than one year. 881.43 R dollars was spent on hours/salary for admission and resignation. This short time of permanence doubled the cost for hiring a new professional. PMID- 16308620 TI - [Nursing staff turnover: a study at a school-hospital]. AB - The goal of this exploratory-descriptive study was to measure and compare nursing staff turnover levels at the University Hospital of Sao Paulo University. The population consisted of 255 nursing workers - 78 nurses, 20 technicians and 156 auxiliaries - who resigned from the hospital between 1997 and 2000. Turnover levels were measured by means of global indicators, admission and resignation, net replacement on the job and variation rates. Quantitative data analysis indicated a low nursing staff turnover rate at this hospital. PMID- 16308621 TI - [Tasks performed by nurses at inpatient units in a training hospital]. AB - The purpose of this study was to identify and analyze tasks performed by nurses at inpatient units in a large general hospital in Brasilia, Federal District, which also functions as a training hospital. A quantitative, exploratory, descriptive study was carried out and involved a total of 612 hours of direct observation of the activities performed by 18 nurses at the General Medicine, Surgical, Pediatric and Maternity units. The tasks observed were classified as: administration, nursing care delivery, education and related to information system. Nurses dedicate a large part of their time to administrative tasks, followed by tasks related to nursing care delivery and information system, performing almost no educative tasks. PMID- 16308622 TI - [Factors favoring the participation of aged inpatients' caregivers in care]. AB - This study aimed to identify and analyze what factors favor the participation of aged inpatients' caregivers in care during the hospitalization period, according to relatives and nursing group members responsible for care delivery. Thirty aged inpatients' relatives and 30 nursing group members were interviewed at a hospital in the interior of Sao Paulo state. Thematic analysis of the answers revealed that interpersonal relationships, interest in taking part in care activities and previous knowledge are the most favorable factors contributing to family members' participation in care. PMID- 16308623 TI - [Quality of life in chronic kidney failure patients receiving hemodialysis treatment]. AB - This descriptive study aims to evaluate the quality of life in patients receiving hemodialysis (HD) treatment and to identify the daily activities that may impair their quality of life. We investigated 125 chronic kidney failure patients under hemodialysis treatment by means of the Medical Outcome Survey-Short-Form 36 (SF 36), and their daily activities, by means of semi-structured interviews. For statistical data analysis, a significance level of 5% was used. The results evidenced that these individuals' quality of life is impaired, with lower scores for physical, emotion and vitality aspects. There was a negative correlation between time spent on HD and the physical component (r= - 0.75) and between hemodialysis time and daily activities such as work, housework and practical activities. Physical and leisure activities were the most affected in the general sample as well as in the one stratified by gender. PMID- 16308624 TI - [Oncological care according Alfred Schutz]. AB - The study was realized among oncological nurses in their daily work routine and aimed to understand these professionals' subjective action, starting from their relation with patients, adopting a phenomenological reference framework based on the ideas of Alfred Schutz. The question: what does working in oncological care mean to you? Please describe, was used to collect statements, which were analyzed and clarified the typical action of a nurse caregiver in this daily routine. The study revealed that oncological care implies dealing with humans in a fragile situation; requires a relationship of affectivity; is care delivery that entails the genesis of professional burnout. Care delivery in oncology is highly complex, requiring a professional competence that goes beyond the technical-scientific sphere. Nursing professionals need to seek strategies which enable them to face the fatigue they are submitted to in their work. PMID- 16308625 TI - [Characterization of patients in a temporomandibular dysfunction and orofacial pain outpatient clinic]. AB - This study aims to describe the characteristics of patients at a temporomandibular dysfunction and orofacial pain outpatient clinic and to discuss the insertion of nursing care in this service. A questionnaire based on the steps of the Nursing Process was applied to a sample of 150 patients attended from May to August 2003. Patients from both genders and aged between 12 and 77 years old were sequentially admitted to the study. Population data revealed that a majority was female (85%); the predominant age was from 21 to 60 years old (76%); only 3% did not present any formal instruction. The nursing role introduced in this multidisciplinary clinic created conditions to assess demographic and epidemiologic data, identify user needs and develop self-care abilities and attitudes. The proposed model made is possible to organize data collection and promote research. PMID- 16308626 TI - [Pain intensity of pain and adequacy of analgesia]. AB - An unprecedented study in Brazil analyzed pain intensity and adequacy of analgesia at an emergency center. One hundred accident victims attended at a trauma reference hospital were evaluated. Pain was present in 90% of cases; 56.0% complained of severe pain on first evaluation and, three hours later, 26.0% remained in severe pain and 38.0% in moderate pain. A significant portion of the study population did not receive analgesia during the observation period. Considerable numbers of inadequate analgesia were found, confirming the reduced importance of analgesia in trauma in Brazil. PMID- 16308627 TI - [What drugs users say and do not say, using experiences and the projective technique]. AB - This study aimed to get to know some visible and hidden meanings in drugs users' experience. The interviews were associated with the Projective Technique (free and thematic drawing), based on which we analyzed the subjective content and what was expressed by the users. The results demonstrated the existence of family deterioration, stigma and prejudice. PMID- 16308628 TI - [Labor accidents involving the eyes: assessment of occupational risks involving nursing workers]. AB - The study aimed at identifying nursing workers who were victims of eye accidents and the type of accident; describing the measures taken and proposing Health Education methods. A descriptive and exploratory study was carried out at a public maternity hospital from September 2002 to January 2003. Data were collected through direct observation of the environment and interviews with workers. Subjects were ten professionals (one nurse, two technicians and seven nursing auxiliaries) who were victims of work accidents involving the eye. The accidents were grouped according to the type of material that caused the trauma: chemical substances (4), medication (3), mechanical trauma (1), scalp (1) and urine (1). The results reveal that hospital workers are vulnerable to labor accidents because the environment presents biological, chemical and physical risks. An important step to prevent the occurrence of new accidents would be the prevention of human mistakes through permanent training and the use of protection glasses. PMID- 16308629 TI - [The use of videotaping in qualitative research]. AB - Considering advancements in data collection methods, we explore the use of videotaping in qualitative research. This bibliographical study aims at developing reflections on the possibilities of using videotapes in research and at providing material to researchers. The video is used as an instrument of data collection and generation. We mention technical aspects, such as the utilization of a mobile or fixed camera. By means of the latter, the authors report their experience, emphasizing compliance with the neutrality principle and the possibility of editing the images obtained as a means of generating new data. The authors highlight that it was possible to detect contradictions between discourse and behavior through the use of videotaping and interviews. The authors also discuss the ethical principles set by CNS Resolution 196/96 and other ethical questions, whose solutions should be based on the agreement between researchers and subjects. PMID- 16308630 TI - [The concept of communicative action: a contribution to nursing consultation]. AB - Nurses often adopt a mechanist view, perceiving the individual from a biological standpoint and giving little attention to psychological, historical and cultural aspects. This evidences the need for a more humane approach, in which the communication between nurses and health service users can be achieved with more comprehension and participation. The aim of this paper is to introduce the concepts of language and communication in nursing consultation, using as a theoretical and conceptual framework Jurgen Habermas' theory of universal pragmatism. In nurses' everyday routine, it is important to intermediate and adapt knowledge and technical skills with a view to effective performance in humanized care for individuals and the community. The dialogic practice, through reflexive and participative communicative action by nurses, is needed for more humane care. PMID- 16308631 TI - [Politicity of care as an emancipatory reference for nursing: getting to know to care better, delivering care to confront, delivering care to emancipate]. AB - Politicity of care deals with the intelligent and disruptive handling of help power. Such notion can mainly be expressed by the trihedron getting to know to care better, delivering care to confront, delivering care to emancipate which, when adapted to the work process in health, may become emancipatory. This theoretical-philosophical reflection explores the ambiguous aspect of care, expressed by being solidary and becoming political. OBJECTIVES: a- to reflect about the politicity of care, as management of help-power, which is capable of emancipatory subversion; b- to discuss the way capitalist societies deal with health care; c- to point out disruptive dynamics to nurses, based on the emancipatory trihedron of care. The main idea of this article is to indicate applicabilities for the conception of politicity of care in the sphere of the nursing work process, signaling emancipatory potentialities. PMID- 16308632 TI - [Some considerations about nurses' use of non traditional therapies in psychiatric nursing care]. AB - In this work, a revision of the literature was carried out to identify articles that described non traditional therapeutic modalities psychiatric nurses can use in their daily practice. The described modalities were Music, Motor Activity, Therapeutic Accompaniment and Yoga. Music is able to reconstruct identities, integrate people, reduce anxiety and provide the construction of positive self esteem. Motor Activity increases self-esteem, reduces inactivity and increases the participation of the psychiatric patient in other activities. In Therapeutic Accompaniment, nurses follow patients in daily routine with a view to their reintegration in the different situations they have to face. Yoga allows for improved memory, reduces emotional tension, depression, anxiety and irritability, and promotes relaxation and a greater feeling of self-control. All activities should be integrated in a global therapeutic plan for each user and be part of daily service programming. PMID- 16308633 TI - [Immunogenetic factors associated with type 1 diabetes mellitus]. AB - Type 1 diabetes mellitus has been considered an organ-specific autoimmune disease derived from the selective destruction of pancreatic beta cells. It presents a complex pathogenesis, involving the participation of several factors, including the immunogenetic susceptibility with strong association to histocompatibility genes (HLA), environmental events and autoimmune response with the presence of autoantibodies and/or autoreactive lymphocytes, culminating in metabolic abnormalities. In this study, the literature review describes mechanisms through which some factors cause susceptibility to its appearance and, additionally, prediction innovations regarding this disorder, which will certainly contribute to nursing care for patients with type 1 diabetes. PMID- 16308634 TI - [Theorizing about systems: an ecological task for patient safety research]. AB - As a global safety movement gathers momentum, experts call for a "systems approach" to improve the safety of today's health care environments. Yet, what kinds of systems theories should guide the field of patient safety research? In this paper, it is argued that nurses and other health professionals can use theory and principles from the field of ecological restoration, which is the repair of damaged ecosystems, to study and strengthen the safety of health care environments around the world. When we use restoration science to theorize about health care systems, we develop the ability to think ecologically about our relations with each other and with the environments we share. As we integrate knowledge of restoration science with nurses' knowledge and other knowledge in health care, we may actually create safer health care systems for all at a human and material cost that we are able and willing to pay. PMID- 16308635 TI - [The development of handbooks of health care guidelines]. AB - This article is an experiment report on the author's development of health care guideline manuals, focusing on a methodological description. These guiding manuals aim to support health care professionals to provide oral teaching to patients and family members, thus enhancing health education. PMID- 16308636 TI - The trend towards a conservative approach in the surgical management of breast cancer. PMID- 16308638 TI - Do no harm: do thyself no harm. PMID- 16308639 TI - Biostatistics 308. Structural equation modeling. PMID- 16308640 TI - Clinical practice guidelines. AB - This paper introduces the concepts of evidence-based clinical practice guidelines. It describes the key elements of guideline development, using examples from the Scottish Intercollegiate Guidelines Network (SIGN), and then goes on to discuss how practitioners in Singapore and other countries can find and use guidelines from other areas of the world. It concludes with a short section on the future direction of guideline development. PMID- 16308641 TI - Breast lymphoscintigraphy for sentinel node identification in breast cancers with clinically-negative axillary nodes. AB - INTRODUCTION: To analyse and determine the clinical value of lymphoscintigraphy for sentinel lymph node (SLN) localisation in woman undergoing surgery for breast cancer, and evaluate the predictive value of SLN versus axillary lymph node (ALN) status in these patients. METHODS: Preoperative breast lymphoscintigraphy was performed in 35 female patients with breast cancer and clinically-negative ALNs. The mean age was 52.8 years (age range 38 to 73 years). The lymphoscintigraphy was performed using 74 MBq of Tc-99m nanocolloid subdermal injection over the tumour. The SLN location was marked on the skin. All patients underwent standard modified radical mastectomy with axillary lymph node dissection (ALND). A comparison of SLN and ALN histopathological results was completed in order to define the means by which the SLN biopsy was able to reflect the final status of ALNs. RESULTS: In 20/35 (57.1 percent) cases, SLNs were visualised in 20-minute dynamic imaging. In 12 patients, SLNs were seen after delayed imaging and/or by repositioning the patient. Overall, the estimated SLN identification rate was 91.4 percent. Of 32 patients in whom SLNs were localised by lymphoscintigraphy, nine were positive for metastatic tumours and the rest were negative for tumour involvement. In four of these nine patients, SLN was the only node that contained metastatic tumour cells while in five patients, an additional concomitant ALN metastasis was detected. In four patients, SLN was negative on frozen section, but skip ALN metastases were noted. Of three patients in which SLNs were not localised by lymphoscintigraphy, two had positive ALNs for tumour cells and the remaining one was negative for tumour involvement. CONCLUSION: We concluded that SLN localisation using lymphoscintigraphy is an accurate minimally-invasive procedure for staging breast cancer patients with clinically-negative ALNs, and can substantially reduce the morbidity and costs of surgical treatment by avoiding unnecessary ALND in the majority of patients. PMID- 16308642 TI - Predictors of axillary lymph node metastases in women with early breast cancer in Singapore. AB - INTRODUCTION: The presence of axillary lymph node metastases is an important prognostic factor in breast cancer. Sentinel lymph node biopsy (SLNB) is an emerging method for the staging of the axilla. It is hoped that with SLNB, the morbidity from axillary lymph node dissection (ALND) can be avoided without compromising the staging and management of early breast cancer. However, only patients found to be SLNB negative benefit from this procedure, as those with positive SLNB may still require ALND. Our objective is to study the various clinico-pathological factors to find predictive factors for axillary lymph node involvement in early breast cancer. It is hoped that with these factors, we will be better able to identify groups of patients most likely to benefit from SLNB. METHODS: A retrospective study of 380 early breast cancer cases (stage T1 and T2, N0, N1, M0) in women treated in the Department of General Surgery, Tan Tock Seng Hospital, between January 1999 and August 2002, was conducted. Incidence of nodal metastases was correlated with clinico-pathological factors, and analysed by univariate and multivariate analyses. RESULTS: Approximately 35 percent of the 380 cases of early breast cancer had nodal metastases. Multivariate analyses revealed four independent predictors of node positivity: tumour size (p-value equals 0.0001), presence of lymphovascular invasion (p-value is less than 0.0001), tumours with histology other than invasive ductal or lobular carcinoma (p-value equals 0.04), and presence of progesterone receptors (p-value equals 0.05). CONCLUSION: We have found independent preoperative predictive factors in our local population for the presence of nodal metastases. This information can aid patient selection for SLNB and improve patient counselling. PMID- 16308643 TI - Risk factors of breast cancer in women in Kelantan, Malaysia. AB - INTRODUCTION: Breast cancer is the commonest cancer for females in Malaysia, with a prevalence of 86.2 per 100,000 women in 1996. Breast cancer mortality rate shows an increasing trend in Malaysia from 0.61 in 1983 to 1.8 per 100,000 women in 1992. The aim of this study was to identify the risk factors for breast cancer in women in Malaysia. METHODS: A matched case-control study was carried out at referral hospitals in Kelantan from July 2000 to June 2001. A standardised questionnaire that comprised socio-demographical factors, obstetrical and gynaecological histories, anthropometric measurements, and other potential risk factors for breast cancer, was used to interview 147 histologically-confirmed breast cancer patients and 147 controls. Controls were non-breast cancer patients who were matched for age and ethnicity, and excluded those with malignancies, or having gynaecological, hormonal or endocrine problems. Simple and multiple conditional logistic regressions were used for analyses. RESULTS: Factors contributing toward increased risk of breast cancer were nulliparity (odds ratio [OR] of 15.3; 95 percent confidence interval [CI] of 3.2, 72.4), overweight (OR of 2.1; 95 percent CI of 1.1, 3.9), family history of breast cancer (OR of 4.3; 95 percent CI of 1.3, 14.1) and previous use of oral contraceptives (OR of 2.5; 95 percent CI of 1.3, 4.8). CONCLUSION: This study reconfirmed that similar risk factors identified in Western populations were responsible for the occurrence of breast cancer in Kelantan. It also supported the theory that breast cancer occurrence was related to oestrogen exposure and familial factors. It suggested the importance of having children, maintaining ideal body weight and caution for oral contraceptive users and women with a family history of breast cancer. PMID- 16308644 TI - Atypical and suspicious categories in fine needle aspiration cytology of the breast: histological and mammographical correlation and clinical significance. AB - INTRODUCTION: This study aims to correlate fine-needle aspiration specimens diagnosed as C3 (atypical, probably benign) and C4 (suspicious, probably malignant) with histology and mammography, and to evaluate these two cytology categories in terms of diagnostic usefulness and patient management. METHODS: All fine-needle aspiration (FNA) specimens in categories C3 or C4 at the Maharaj Nakorn Chiang Mai Hospital, Thailand between 2000-2004 were reviewed. Results were correlated with available histological and mammographical studies. RESULTS: 148 FNA specimens were identified, comprising 43 category C3 and 105 category C4. Histology was available in 90 cases. 14 (64 percent) C3 cases showed benign histology on biopsy and eight (36 percent) were malignant. 13 (19 percent) C4 cases were benign on biopsy, whereas 55 (81 percent) were malignant. Mammographical studies were available in 56 of the histologically-proven cases. All seven cases with benign mammograms had benign histology, and all 26 cases called "highly suggestive of malignancy" were malignant on histology (five C3 and 21 C4). Of the 23 cases called "suspicious abnormality" on mammography, 14 turned out to be malignant on biopsy (one C3 and 13 C4). CONCLUSION: Our study supports maintaining cytology categories C3 and C4. About two-thirds of C3 cases were benign on biopsy whereas 81 percent of C4 cases were malignant (p-value is less than 0.001). There was complete correlation between histological and mammographical studies except those with equivocal mammograms. Our study supports the combined use of clinical, mammographical and cytological findings for optimal patient management. This is especially important for patients with C3 aspiration results, in order to avoid unnecessary surgery for benign lesions. PMID- 16308645 TI - Prognostic value of an index for serum globulin compensation in colon and breast cancers. AB - INTRODUCTION: Decreased serum albumin (SA) levels have been used extensively as prognostic indicators in many chronic debilitating diseases. The decrease may be partly compensated by globular proteins. The failure of globulins to compensate may reflect advanced disease. We examined the prognostic value of the level of serum globulins in colorectal and breast cancers. METHODS: Data of 80 patients with advanced colon and breast cancers were analysed. Of these, 46 patients died within six months of measurement of their serum proteins, and the rest were followed-up for more than six months after measurements of their serum proteins were taken. A mathematical formula, representing the globulin compensation index (GCI), was recently developed from the measured SA levels and globulins. Patients were then classified into three categories: negative GCI and negative compensation; GCI of 0 to less than 1.0 with partial compensation; and GCI equal or greater than 1.0 with full compensation. RESULTS: Among the deceased patients, 45.7 percent had negative GCI, compared to 26.5 percent of patients in the survivors group. For partial compensation, 30.4 percent of patients were from the deceased group, and 32.4 percent were from the survivors group. For full compensation (elevated GCI), 23.9 percent of patients were from the deceased group, compared to 41.1 percent from the survivors group (p-value equals 0.031). CONCLUSION: Patients with low GCI are more likely to have bad prognoses, whereas those with higher GCI have more favourable prognoses. Globulin compensation may be a reliable prognostic factor in advanced colorectal and breast cancers, and possibly in other chronic illnesses. The GCI may serve as a useful tool in the measurement of this compensation. PMID- 16308646 TI - Serious cutaneous adverse reactions to traditional Chinese medicines. AB - INTRODUCTION: Serious cutaneous adverse reactions to traditional medicines are not well described or reported in the literature, despite growing use of these medicines. METHODS: This is a case series of four patients who were found to have various serious cutaneous adverse reactions to the traditional Chinese medicines that they had taken. RESULTS: In this series, there was a patient with toxic epidermal necrolysis from traditional Chinese medicine, another with acute generalised exanthematous pustulosis from piroxicam and salicylate-contaminated traditional Chinese medicine, and two patients with drug hypersensitivity syndrome--one from traditional Chinese medicine and the other from phenylbutazone adulterated traditional Chinese medicine. CONCLUSION: The series illustrates that serious cutaneous adverse reactions do occur with traditional medicines and emphasises the importance of being aware of such reactions. PMID- 16308647 TI - Semi-constrained total elbow arthroplasty for the treatment of rheumatoid arthritis of the elbow. AB - INTRODUCTION: We retrospectively reviewed the results of total elbow arthroplasty in patients with rheumatoid arthritis of the elbow followed for a minimum of two years. METHODS: Between September 1999 and March 2001, seven patients with rheumatoid arthritis of the elbow were treated with total elbow arthroplasty using the semi-constrained Coonrad-Morrey elbow replacement prostheses. One patient died 19 months after her surgery and was excluded from the study. Two patients had bilateral total elbow replacements. Eight elbows were thus available for review. The mean age of our patients at the time of surgery was 55.5 years. The indication for surgery was severe pain and stiffness in six elbows, and distal humerus fractures in two elbows. RESULTS: The mean duration of postoperative hospitalisation stay was 6.6 days. The average length of follow-up was 39.4 months. Six elbows had no pain, while two elbows had mild pain. The mean arc of flexion was 101.3 degrees. The mean Mayo elbow performance score was 93.1 points. Excellent results were achieved in six elbows, while two elbows had good outcome. There was one case of intraoperative lateral condylar fracture treated with internal fixation. One patient developed blisters postoperatively but resolved with dressings and antibiotics. CONCLUSION: Our small study revealed good to excellent short-term outcome with the use of semi-constrained total elbow arthroplasty for the treatment of rheumatoid arthritis of the elbow in Asian patients. PMID- 16308648 TI - Successful management of a bleeding duodenal varix by endoscopic banding. AB - Endoscopic treatment of bleeding duodenal varices is less invasive than the usual surgical options. However, there is limited experience with endoscopic treatment of bleeding duodenal varices, especially with that of endoscopic ligation. We report a 55-year-old man with a bleeding duodenal varix that was successfully ligated endoscopically. He has been followed up for nine months with no recurrence of bleeding. Endoscopic ligation may offer a new and effective treatment modality for bleeding duodenal varix. PMID- 16308649 TI - Reversible splenial lesion in clinically mild encephalitis. AB - Clinically mild encephalitis with a reversible lesion in the central splenium of the corpus callosum (SCC) is a recently-described clinicoradiological entity. We report a 20-year-old man presenting with fever and a single episode of generalised seizures. Initial magnetic resonance (MR) images showed an ovoid lesion with T1 and T2 signal prolongation, restricted diffusion and decreased apparent diffusion coefficient values in the centre of the SCC, which resolved completely on a repeat MR imaging done three months later. Clinically, the patient had a mild clinical course and made a full recovery. This clinicoradiological entity with an excellent prognosis is elaborated with possible differential diagnoses given. Emphasis is placed on avoiding unnecessary invasive investigation or therapeutic intervention. PMID- 16308650 TI - Cerebral tuberculosis mimicking intracranial tumour. AB - Cerebral tuberculoma is a rare entity and is one of the causes of intracerebral mass lesions. A rapid diagnosis based on pathological findings improves its prognosis. We describe two cases where the tuberculoma was located in the cavernous sinus and prepontine cistern, respectively. The first case was a 36 year-old man who was admitted with progressive headache, left ptosis and diplopia. Computed tomography showed a solid enhancing mass in the left cavernous sinus. Diagnosis of meningioma was proposed and a left pterional craniotomy was performed. Histopathological examination revealed granulomatous inflammation with areas of caseation necrosis. The second case was a 20-year-old man who presented with headache, new-onset strabismus, diplopia, malaise, weight loss and low-grade fever. The lesion mimicked an aggressive meningioma on imaging. The patient was operated for primary diagnosis of cerebral tumour. The histopathological examination of the excised lesion revealed a tuberculoma. Although the incidence of tuberculosis is decreasing, a high index of suspicion must be maintained for the diagnosis of intracranial masses in the presence of risk factors for tuberculosis. PMID- 16308652 TI - Hypertension in patients with type 2 diabetes mellitus: management needs to be more intensified. PMID- 16308653 TI - Are the World Health Organisation case definitions for severe acute respiratory syndrome sufficient at initial assessment? PMID- 16308654 TI - Darwin: fairly good for a medical school dropout. PMID- 16308655 TI - Charles Darwin: history's most famous naturalist. PMID- 16308656 TI - Charles Darwin: history's most famous naturalist. PMID- 16308660 TI - BBS8 is rarely mutated in a cohort of 128 Bardet-Biedl syndrome families. AB - BBS8 is one of the eight genes identified to date for Bardet-Biedl syndrome (BBS) an autosomal recessive condition associated with retinitis pigmentosa, obesity, polydactyly, cognitive impairment and kidney failure. The identification of BBS8 gave the key to the pathogenesis of the condition as a primary ciliary disorder. To date, only three families mutated in the BBS8 gene have been reported. Here, we report on three additional families with BBS8 mutations from a series of 128 BBS families. Two of the three families have homozygous mutations and one has a heterozygous mutation. Mutations in BBS8 probably account for only a minority of BBS families (2%), underlining the difficulty of genotyping heterogeneous conditions. PMID- 16308661 TI - Failure to decrease blood pressure during sleep: non-dippers are among us. PMID- 16308663 TI - Anatomical study of the communicating branches between the medial and lateral plantar nerves. AB - The plantar areas of the foot have specific biomechanical characteristics and play a distinct role in balance and standing. For the forefoot surgeon, knowledge of the variations in the anatomy of communicating branches is important for plantar reconstruction, local injection therapy and an excision of interdigital neuroma. The anatomy of the communicating branches of the plantar nerves between the fourth and third common plantar digital nerves in the foot were studied in 50 adult men cadaveric feet. A communicating branch was present between the third and fourth intermetatarsal spaces nerves in all eight left feet and in six right feet (overall, 28%), and absent in 36 (72%). A communicating branch was found in 14 ft. Ten of the 14 communications were from the lateral to the medial plantar nerve. The length of the communicating branch ranged from 8 to 56 mm (average 16.4 mm) and its diameter was 0.2-0.6 times of the fourth common plantar digital nerve. The angle of the communicating branch with the common plantar digital nerve from which it originated was less than 30 degrees in 11 ft, 30-59 degrees in 27 ft, 60-80 degrees in 8 ft, and more than 80 degrees in 4 ft. Classification of the branch is based on the branching pattern of the communicating branch and explains variations in plantar sensory innervations. We think that the perpendicular coursing communicating branch is at higher risk to be severed during surgery. PMID- 16308662 TI - Protective effect of 1,2,4-benzenetriol on LPS-induced NO production by BV2 microglial cells. AB - Hydroxyhydroquinone or 1,2,4-benzenetriol (BT) detected in the beverages has a structure that coincides with the water-soluble form of a sesame lignan, sesamol. We previously showed that sesame antioxidants had neuroprotective abilities due to their antioxidant properties and/or inducible nitric oxide synthase (iNOS) inhibition. However, studies show that BT can induce DNA damage through the generation of reactive oxygen species (ROS). Therefore, we were interested to investigate the neuroprotective effect of BT in vitro and in vivo. The results showed that instead of enhancing free radical generation, BT dose-dependently (10 100 microM) attenuated nitrite production, iNOS mRNA and protein expression in lipopolysaccharide (LPS)-stimulated murine BV-2 microglia. BT significantly reduced LPS-induced NF-kappaB and p38 MAPK activation. It also significantly reduced the generation of ROS in H2O2-induced BV-2 cells and in H2O2-cellfree conditions. The neuroprotective effect of BT was further demonstrated in the focal cerebral ischemia model of Sprague-Dawley rat. Taken together, the inhibition of LPS-induced nitrite production might be due to the suppression of NF-kappaB, p38 MAPK signal pathway and the ROS scavenging effect. These effects might help to protect neurons from the ischemic injury. PMID- 16308666 TI - Isolated pulmonary hypertension secondary to rheumatoid arthritis. AB - The authors report a case of a woman with pulmonary hypertension secondary to rheumatoid arthritis, whose treatment with azathioprine resulted in normalization of pulmonary artery pressure and resolution of clinical symptoms. Different etiologies for pulmonary hypertension are discussed and literature review is presented. PMID- 16308665 TI - Extra- and intramuscular nerve supply of the muscles of the anterior antebrachial compartment: applications for selective neurotomy and for botulinum toxin injection. AB - Hypertonia of the upper limb due to spasticity causes pronation of the forearm and flexion of wrist and fingers. Nowadays this spasticity is often treated with injections of botulinum toxin and sometimes with selective fascicular neurotomy. To correctly perform this microsurgical technique, it is necessary to get precise knowledge of the extramuscular nerve branching in order to be better able to select the motor branches which supply the muscles involved in spasticity. The same knowledge is required for botulinum toxin injections which must be made as near as possible to the zones where intramuscular nerve endings are the densest, which is also where neuromuscular junctions are the most numerous. Thus, it is necessary to better know these zones, but their knowledge remains today imprecise. The muscles of the anterior compartment of 30 forearms were dissected, first macroscopically, then microscopically, to study the extra- and intramuscular nerve supply and the distribution of terminal nerve ramifications. The results were then linked to surface topographical landmarks to indicate the precise location of motor branches for each muscle with the aim of proposing appropriate surgical approaches for selective neurotomies. Then for each muscle, the zones with the highest density of nerve endings were divided into segments, thus determining the optimal zones for botulinim toxin injections. PMID- 16308664 TI - Relationship between ultrastructure and biomechanical properties of the knee meniscus. AB - The purpose of this study was to determine the biomechanical properties of the knee meniscus and to relate them to its ultrastructure. The knee joint menisci are semicircular, fibrocartilaginous structures interposed between the femoral and tibial condyles. For a long time, they were considered to be embryologic vestiges. This study describes the response of the knee joint meniscus to circumferential, radial and axial compressive forces. The results show an anisotropic response of the knee joint meniscus to unconfined compression. The Young's modulus increased approximately twofold between vertical and circumferential or radial directions with a 10 mm/min-compression rate. This response is probably a direct consequence of the orientation of collagen fibres. PMID- 16308667 TI - Acute dermatomyositis with subcutaneous generalized edema. AB - The authors report a 40-year-old Caucasian man with relapsing muscle and skin involvement of dermatomyositis treated with high-dose corticosteroids, taken orally, and methotrexate and human gamma globulin, both administered intravenously. After 4 months of aggressive treatment, he presented with generalized edema, considered secondary to dermatomyositis. Aggressive immunosuppression did not stop disease progression. The literature concerning anasarca due to inflammatory myopathies is revised. PMID- 16308668 TI - Pharmacokinetics/pharmacodynamics of nondepleting anti-CD4 monoclonal antibody (TRX1) in healthy human volunteers. AB - PURPOSE: TRX1 is a nondepleting anti-CD4 monoclonal IgG1 antibody being developed to induce tolerance by blocking CD4-mediated functions. The purpose of this study is to describe the pharmacokinetics (PK) and pharmacodynamics (PD) of TRX1 and to develop a receptor-mediated PK/PD model that characterizes the relationships between serum TRX1 concentration and total and free CD4 expression in healthy male volunteers. METHODS: Nine subjects from three dosing cohorts in double blinded, placebo-controlled phase I clinical study was included in the analysis. Serum TRX1 levels were determined using enzyme-linked immunosorbent assay. Blood total and free CD4 receptor levels were determined by using flow cytometric analyses. The receptor-mediated PK/PD model was developed to describe the dynamic interaction of TRX1 binding with CD4 receptors. RESULTS AND CONCLUSIONS: TRX1 displayed nonlinear pharmacokinetic behavior and the CD4 receptors on T cells were saturated and down-modulated following treatment with TRX1. Results from in vitro studies using purified human T cells suggested that CD4-mediated internalization may constitute one pathway by which CD4 is down-modulated and TRX1 is cleared in vivo. The developed receptor-mediated PK/PD model adequately described the data. This PK/PD model was used to simulate PK/PD time profiles after different dosing regimens to help guide the dose selection in future clinical studies. PMID- 16308669 TI - Prediction of milk/plasma drug concentration (M/P) ratio using support vector machine (SVM) method. AB - PURPOSE: Development of reliable computational models to predict/classify milk-to plasma (M/P) drug concentration ratio remains a challenging object. Support vector machine (SVM) method, as a new algorithm, was constructed to distinguish the potential risk of drugs to nursing infants. METHODS: Each drug was represented by a large pool of descriptors, of which five were found to be most important for constructing the predictive models. Next, two classification models, linear discriminant analysis (LDA) and SVM, were developed with bootstrapping validation based on the selected molecular descriptors. RESULTS AND CONCLUSIONS: The classification accuracy of training set and test set for SVM was 90.63 and 90.00%, respectively. The total accuracy for SVM was 90.48%, which was higher than that of LDA (77.78%). Comparison of the two methods shows that the performance of SVM was better than that of LDA, which implies that the SVM method is an effective tool in evaluating the risk of drugs when experimental M/P ratios have not been investigated. PMID- 16308670 TI - Microinfusion using hollow microneedles. AB - PURPOSE: The aim of the study is to determine the effect of experimental parameters on microinfusion through hollow microneedles into skin to optimize drug delivery protocols and identify rate-limiting barriers to flow. METHODS: Glass microneedles were inserted to a depth of 720-1080 microm into human cadaver skin to microinfuse sulforhodamine solution at constant pressure. Flow rate was determined as a function of experimental parameters, such as microneedle insertion and retraction distance, infusion pressure, microneedle tip geometry, presence of hyaluronidase, and time. RESULTS: Single microneedles inserted into skin without retraction were able to infuse sulforhodamine solution into the skin at flow rates of 15-96 microl/h. Partial retraction of microneedles increased flow rate up to 11.6-fold. Infusion flow rate was also increased by greater insertion depth, larger infusion pressure, use of a beveled microneedle tip, and the presence of hyaluronidase such that flow rates ranging from 21 to 1130 microl/h were achieved. These effects can be explained by removing or overcoming the large flow resistance imposed by dense dermal tissue, compressed during microneedle insertion, which blocks flow from the needle tip. CONCLUSIONS: By partially retracting microneedles after insertion and other methods to overcome flow resistance of dense dermal tissue, protocols can be designed for hollow microneedles to microinfuse fluid at therapeutically relevant rates. PMID- 16308671 TI - Analytical studies on the prediction of photosensitive/phototoxic potential of pharmaceutical substances. AB - PURPOSE: Phototoxic responses after administration of photosensitive pharmaceutics have been recognized as undesirable side effects, and predicting potential hazardous side effects is gaining importance as new drugs are introduced to the market. In this work, we characterize the photochemical/photobiological properties of model compounds to develop an effective screening method for the prediction of phototoxic/photosensitive potential. METHODS: Twenty-one known photosensitive/phototoxic compounds and five weak/nonphototoxic compounds were subjected to ultraviolet (UV) spectral analyses and photochemical evaluation including the determination of produced reactive oxygen species (ROS) and photostability study. The photooxidation of linoleic acid was also monitored in the presence of tested compounds, guided on the formation of thiobarbituric acid reactive substances. RESULTS: Most photosensitive/phototoxic drugs tested, even weak UV absorbers, at a concentration of 200 microM showed significant production of ROS under 18 h light exposure (30,000 lx). On the other hand, ROS generated from weak/nonphototoxic compounds, including strong UV absorber benzocaine, were low or negligible. Although exposure of quinine to light resulted in significant degradation (half life, t1/2=6.4 h), it was dramatically attenuated by the addition of ROS scavengers, especially sodium azide (t1/2=122.6 h). Furthermore, concomitant exposure of photosensitive/phototoxic compounds (200 microM) and linoleic acid (1 mM) for 18 h led to the marked formation of lipoperoxide. CONCLUSION: Results indicated that known photosensitive/phototoxic compounds tested have the ability to generate ROS under light exposure, and this photochemical reaction could be associated with their photoinstability and/or phototoxic responses. Based on these findings, determination of ROS, generated from photoirradiated compounds, may be an effective predictive model in recognizing their photosensitive/phototoxic potential. PMID- 16308672 TI - Characterization of the human upper gastrointestinal contents under conditions simulating bioavailability/bioequivalence studies. AB - PURPOSE: This study was conducted to compare the luminal composition of the upper gastrointestinal tract in the fasted and fed states in humans, with a view toward designing in vitro studies to explain/predict food effects on dosage form performance. METHODS: Twenty healthy human subjects received 250 mL water or 500 mL Ensure plus (a complete nutrient drink) through a nasogastric tube and samples were aspirated from the gastric antrum or duodenum for a period up to 3.5 h, depending on location/fluid combination. Samples were analyzed for polyethylene glycol, pH, buffer capacity, osmolality, surface tension, pepsin, total carbohydrates, total protein content, and bile salts. RESULTS: Following Ensure plus administration, gastric pH was elevated, buffer capacity ranged from 14 to 28 mmoL L-1 DeltapH-1 (vs. 7-18 mmol L-1 DeltapH-1), contents were hyperosmolar, gastric pepsin levels doubled, and surface tension was 30% lower than after administration of water. Post- and preprandial duodenal pH values were initially similar, but slowly decreased to 5.2 postprandially, whereas buffer capacity increased from 5.6 mmol L-1 DeltapH-1 (fasted) to 18-30 mmol L-1 DeltapH-1 (p<0.05). Postprandial surface tension in the duodenum decreased by >30%, bile salt levels were two to four times higher, luminal contents were hyperosmotic, and the presence of peptides and sugars was confirmed. CONCLUSIONS: This work shows that, in addition to already well characterized parameters (e.g., pH, and bile salt levels), significant differences in buffer capacity, surface tension, osmolality, and food components are observed pre-/postprandially. These differences should be reflected in test media to predict food effects on intralumenal performance of dosage forms. PMID- 16308673 TI - Dynamics of sulfate-reducing microorganisms (dsrAB genes) in two contrasting mudflats of the Seine estuary (France). AB - By combining molecular biology and biochemical approaches, the dynamics of sulfate-reducing microorganisms (SRM) was investigated in the sediments of the Seine estuary (France). Both intertidal mixing-zone and freshwater mudflats were sampled during a 1-year period; the quantification of SRM was realized by using competitive polymerase chain reaction (PCR) based on dsrAB gene amplification, previously described by Leloup et al. (2004), and sulfate reduction rate (SRR) was determined via the SO4(2-) radiotracer method. Throughout the year, abundance of dsrAB genes and SRR were predominantly high in the top 15 cm of the sediment. A seasonal dynamic was observed; a predominance of activity was noted during the early summer, and seems to be mainly controlled by physical-chemical parameters (temperature and dissolved organic carbon concentration) and topographic evolution of the mudflat (erosion/deposit erosion). PMID- 16308674 TI - Methodological issues in the identification of hip fractures using routine hospital data: a database study. AB - The proportion of the population over the age of retirement has risen in many countries, and this means there is a corresponding rise in the incidence of hip fractures. However, in order to reliably investigate the ability of interventions to prevent fracture, there needs to be a reliable measure of the incidence of hip fracture. The purpose of this study was to examine the inclusion and exclusion criteria used to identify hip fracture from hospital admission data and to examine the impact that these criteria have on estimated incidence of hip fracture. We examine the influence of: individual compared to consultant episode data; primary data compared to any diagnosis of hip fracture; emergency compared to elective admissions; and the influence of type and rate of surgery on incidence estimates. The results showed that classifying hip fractures by use of consultant episodes overestimated the rate of hip fracture by 6-31%, and this overestimation has increased in recent years. The use of primary diagnosis as opposed to any diagnosis underestimates hip fracture by 5%. Two percent (2%) of the people studied had an operation for a hip fracture but did not have a hip fracture diagnosis (many had a multiple fracture diagnosis), and 5.5% of the people studied had an elective admission for the hip fracture (perhaps falling in hospital during an elective admission). We conclude that the selection criteria can have a great influence on the number of hip fractures identified using routine data. There should be a standardized selection procedure for the identification of hip fracture, as this would enable interventions and preventive measures to be evaluated over time and facilitate comparisons of rates in different regions and countries, in order to examine factors associated with hip fracture. PMID- 16308675 TI - Isolation and characterization of Thermus bacteriophages. AB - One-hundred-fifteen bacteriophage strains were isolated from alkaline hot springs in Iceland, New Zealand, Russia (Kamchatka), and the U.S.A. The phages belonged to the Myoviridae, Siphoviridae, Tectiviridae, and Inoviridae families. Over 50% of isolates were isometric or filamentous. One type of siphovirus had giant tails of over 800 nm in length. Phages were further characterized by host range, genome size, DNA restriction endonuclease digestion patterns, and temperature and pH sensitivity. Myoviruses and tectiviruses had a worldwide distribution. Most phages were narrowly host-specific and all were highly resistant against heating and alkaline and acidic pH. This is the first time that tectiviruses and filamentous phages are reported for bacteria of the Thermus-Deinococcus phylum. The presence of tectiviruses, inoviruses, and myoviruses is attributed to acquisition from ancestral gamma-proteobacteria by horizontal gene transfer. PMID- 16308676 TI - [The acromioclavicular joint]. AB - Acute or chronic instability and osteoarthritis of the acromioclavicular (ac) joint may cause significant impairment of the shoulder. In this continuing education report, the pathomorphology of acute ac dislocations is described based on the Rockwood classification. Decision making on conservative or surgical treatment is discussed. Surgical techniques are presented as acromioclavicular or coracoclavicular procedures. Persistent complaints may warrant additional surgical therapy after conservative treatment as well as after primary surgical treatment. In these cases, residual instability must be addressed. A modified Weaver-Dunn procedure is presented in detail. In any patient with shoulder pain, osteoarthritis of the ac joint has to be taken into consideration. Resection of the lateral clavicle has proved to be effective in these patients. PMID- 16308677 TI - [Chronic instability of the acromioclavicular joint. Surgical technique]. PMID- 16308678 TI - [Elastic, stable intramedullary nailing in midclavicular fractures--a change in treatment strategies?]. AB - Midshaft fractures of the clavicle are mostly treated conservatively. For the few cases in which stabilisation is needed, open reduction and plate fixation is the standard procedure. We report a minimally invasive technique of intramedullary stabilisation using a titanium nail carried out in 35 markedly displaced midclavicular fractures. Twelve fractures were simple and 23 consisted of more than two fragments. A total of 62.9% (22/35) of the patients presented additional injuries; 37.1% (13/35) with additional fractures. There were four ipsilateral scapular neck fractures. In 74.3% (26/35) of patients closed reduction was possible, whereas nine fractures required open reduction before the nail inserted from the sternal end of the clavicle could be introduced into the peripheral fragment. All fractures healed clinically and radiologically within 6 weeks. In contrast to conservatively treated fractures, length discrepancies of 1 cm or more, mainly shortening, could be avoided. Using a visual analog scale, significant pain relief (8.4-2.4 points) was already recorded on the first day after surgery. Patients with an isolated fracture of the clavicle showed a nearly normal range of motion only 1 week after surgery, whereas patients with associated scapular or rib fractures needed up to 3 weeks to reach normal shoulder function. No patient complications requiring interventional procedures occurred. Minimally invasive, elastic, stable intramedullary nailing of midshaft fractures of the clavicle should be recommended as an alternative to conservative treatment because of early pain relief, associated rapid functional recovery and anatomical restoration of the clavicle. PMID- 16308679 TI - [Cytokine CXCL13--a possible early CSF marker for neuroborreliosis]. AB - The definitive diagnosis of acute neuroborreliosis (NB) is based upon the presence of lymphomonocytic CSF pleocytosis and intrathecal Borrelia burgdorferi (B.b.)-specific antibody production (expressed by an antibody index of >2). However, the latter might be absent in early stages of the disease. Now a recently discovered additional CSF marker-the cytokine CXCL13-was found to be positive in every initial CSF sample from patients with NB and therefore could be a valuable tool for early diagnosis and initiation of antibiotic therapy. We report an unusual case of NB in a patient with a history of metastatic carcinoma of the prostate and unilateral polyradiculitis. While no intrathecal B.b. specific antibody production could be demonstrated initially, the CSF CXCL13 level was high (>500 ng/g vs <1.7 ng/g in healthy controls). During the course of the disease, the antibody index turned positive (4.8) and the patient responded to antibiotic therapy, thus confirming the diagnosis. In this case, measuring CXCL13 in the CSF would have led to earlier diagnosis and treatment of NB. PMID- 16308680 TI - [The role of philosophy in psychiatry]. AB - Philosophy as a basic study for psychiatry is again attracting interest. Scientific observations in this field are supplementary to empirical science and provide a needed balance to empirical results. In this article we consider the interdisciplinary relationship of both fields and examine its present institutional application. PMID- 16308681 TI - Outcome in severely ill patients with hematological malignancies who received intravenous chemotherapy in the intensive care unit. AB - OBJECTIVE: To assess the outcome in severely ill patients with hematological malignancies who receive intravenous chemotherapy in an intensive care unit (ICU) for a life-threatening malignancy-related complication. DESIGN: Retrospective observational study of prospectively collected data. PATIENTS: All 37 critically ill patients with hematological malignancies who received intravenous chemotherapy in the ICU between January 1997 and March 2005 (mean age 46+/-19 years; mean APACHE II 23+/-7). MEASUREMENTS AND RESULTS: Thirty-seven (69%) patients received chemotherapy because of extensive disease with organ involvement (54%), extensive disease without organ involvement (19%), severe disseminated intravascular coagulation (11%), and other reasons (16%). In 41% there was concomitant infection when chemotherapy was initiated, in 86% a high grade malignancy, and 30% relapsing disease. Twenty-three (62%) patients received mechanical ventilation at the moment of or soon after initiation of chemotherapy for a median duration of 5 days (1-67), and 24% underwent renal replacement therapy during ICU stay. Only ventilation was associated with in-hospital mortality (odds ratio 9.3). ICU, in-hospital, and 6-month mortality rates in nonventilated vs. ventilated patients were 7% and 48%, 14% and 61%, and 54% and 74%, respectively. CONCLUSIONS: Starting chemotherapy in the ICU for a life threatening malignancy related complication can be lifesaving even when infection or organ failure is present. PMID- 16308682 TI - The intensive care support of patients with malignancy: do everything that can be done. PMID- 16308683 TI - Selection of resistance during sequential use of preferential antibiotic classes. AB - OBJECTIVE: To determine the effect of antibiotic class pressure on the susceptibility of bacteria during sequential periods of antibiotic homogeneity. DESIGN AND SETTING: Prospective study in a mixed ICU with three separated subunits of eight, eight, and ten beds. PATIENTS AND PARTICIPANTS: The study examined the 1,721 patients with a length of stay longer than 2 days. INTERVENTIONS: Three different antibiotic regimens were used sequentially over 2 years as first-choice empirical treatment: cephalosporins, fluoroquinolone, or a penicillin-beta-lactamase inhibitor combination. Each regimen was applied for 8 months in each subunits of the ICU, using "latin square" design. RESULTS: We treated 731 infections in 546 patients (32% of patients staying more than 48 h). There were 25.5 ICU-acquired infections per 1,000 patient-days. Infecting pathogens and colonizing bacteria were found in 2,739 samples from 1,666 patients (96.8%). No significant change in global antibiotic susceptibility was observed over time. However, a decrease in the susceptibility of several species was observed for antibiotics used as the first-line therapy in the unit. Selection pressure of antibiotics and occurrence of resistance during treatment was documented within an 8-month rotation period. CONCLUSIONS: Antibiotic use for periods of several months induces bacterial resistance in common pathogens. PMID- 16308684 TI - Reduced stress tolerance of glutamine-deprived human monocytic cells is associated with selective down-regulation of Hsp70 by decreased mRNA stability. AB - In critically ill patients, clinicians observe a reverse correlation of survival and a decreased plasma concentration of the most abundant free amino acid, glutamine (Gln). However, in this context, the role of Gln remains largely elusive. Gln is used as an energy substrate by monocytes. Gln deprivation of these cells results in an increased susceptibility to cell stress and apoptosis, as well as in a reduced responsiveness to pro-inflammatory stimuli. We performed a systematic study to elucidate the molecular mechanism by which Gln depletion affects the heat stress response of the monocytic cell line U937. Proteomic analysis revealed that Gln depletion was associated with specific changes in the protein expression pattern. However, the overall level of tRNA-bound Gln remained unaffected. The stress protein heat shock protein (Hsp) 70 showed the highest reduction in protein synthesis. This was due to enhanced mRNA decay during Gln starvation while the transcriptional and the translational control of Hsp70 expression remained unchanged. A physiological Gln concentration and above was found to be necessary for maximum Hsp70 accumulation upon heat shock. Thus, the study shows a specific link between Gln metabolism and the regulation of heat shock proteins. PMID- 16308685 TI - cIAP2 is highly expressed in Hodgkin-Reed-Sternberg cells and inhibits apoptosis by interfering with constitutively active caspase-3. AB - In this study, the expression of activated caspase-3 by the tumor cells of classical Hodgkin lymphoma (cHL), the Hodgkin-Reed-Sternberg (HRS) cells, is confirmed. This raises the question why caspase-3 does not kill HRS cells. There are only a few molecules, which are able to directly inhibit caspase-3. One of them is cIAP2. We show that cIAP2 is expressed in the HRS cells in 20 of 23 cHL cases by in situ hybridization. Suppression experiments with cIAP2 antisense RNA show that down-regulation of cIAP2 significantly reduces apoptosis resistance in cHL cell lines. cIAP2 overexpression appears to be unique for HRS cells since the tumor cells of non-Hodgkin lymphomas are nearly cIAP2-negative. We demonstrate that cIAP2 is inducible by CD30 stimulation in cHL cell lines of T-cell origin and anaplastic large cell lymphoma cell lines, whereas cHL cell lines of B-cell origin constitutively express cIAP2. Inhibition of cIAP2 expression by cIAP2 antisense RNA decreases resistance to apoptosis. The results indicate that cIAP2 contributes to the apoptosis resistance of HRS cells, mainly by inhibiting effector caspases. According to these findings, a therapeutical application of inhibitors of apoptosis proteins antagonists in cHL appears promising. PMID- 16308686 TI - Effect of SsrA (tmRNA) tagging system on translational regulation in Streptomyces. AB - ssrA genes encoding tmRNA with transfer and messenger RNA functions are ubiquitous in bacteria. In a process called trans-translation, tmRNA enters a stalled ribosome and allows release of the original mRNA, then tmRNA becomes the template for translation of a short tag that signals for proteolytic degradation. We provide here the first evidences that the tmRNA tagging system (ssrA and cohort smpB) is active in Streptomyces. Transcription of the genes was shown and construction of a genetic probe allowed detection of a tmRNA-tagged peptide. Obtention of ssrA and smpB mutants of Streptomyces lividans showed that the ssrA system is dispensable in Streptomyces. Morphologies of the mutants colonies were similar to the wild type, thus tmRNA-mediated tagging does not seem to have, under conditions used, a significant effect in the Streptomyces differentiation. PMID- 16308687 TI - Concordance of thresholds for carcinogenicity of N-nitrosodiethylamine. AB - Three publications on the carcinogenicity of N-nitrosodiethylamine (NDEA) in the livers of F-344 or Wistar rats were examined for concordance of the data. Two reports recorded the appearance of tumors after treatment with NDEA, although one used a different dosing schedule that included phenobarbital promotion. Two studied glutathione S-transferase-placental positive (GST-p+) foci in liver at several doses. One also analyzed DNA for adducts from NDEA. This analysis revealed that when the dose was calculated in molecules/kg/day, the thresholds for the incidence of liver tumors were different by about 1.5 orders of magnitude. But when the dose was calculated as the total cumulative dose, the thresholds for tumor appearance (about 10(20.3) molecules/kg) were in agreement within the error of calculation. Combining the data for GST-p+ foci revealed remarkable agreement between the two reports and a threshold for the appearance of these foci at about 10(19.5) molecules/kg of total cumulative dose of NDEA. DNA adducts fit an exponential curve better than a linear. GST-p+ foci and adducts from NDEA were observed at doses below the threshold dose for the appearance of tumors. These results suggest that: cumulative dose is a better metric than daily dose and that adducts and GST-p+ foci appear at doses below those at which tumors appear. These results further support the observations of the authors that thresholds for carcinogenicity of this genotoxic carcinogen exist and that adducts and altered foci appear at lower doses than the threshold for carcinogenicity. PMID- 16308688 TI - Manual tracking in three dimensions. AB - Little is known about the manual tracking of targets that move in three dimensions. In the present study, human subjects followed, with the tip of a hand held pen, a virtual target moving four times (period 5 s) around a novel, unseen path. Two basic types of target paths were used: a peanut-shaped Cassini ellipse and a quasi-spherical shape where four connected semicircles lay in orthogonal planes. The quasi-spherical shape was presented in three different sizes, and the Cassini shape was varied in spatial orientation and by folding it along one of the three bend axes. During the first cycle of Cassini shapes, the hand lagged behind the target by about 150 ms on average, which decreased to 100 ms during the last three cycles. Tracking performance gradually improved during the first 3 s of the first cycle and then stabilized. Tracking was especially good during the smooth, planar sections of the shapes, and time lag was significantly shorter when the tracking of a low-frequency component was compared to performance at a higher frequency (-88 ms at 0.2 Hz vs. -101 ms at 0.6 Hz). Even after the appropriate adjustment of the virtual target path to a virtual shape tracing condition, tracking in depth was poor compared to tracking in the frontal plane, resulting in a flattening of the hand path. In contrast to previous studies where target trajectories were linear or sinusoidal, these complex trajectories may have involved estimation of the overall shape, as well as prediction of target velocity. PMID- 16308689 TI - Orienting and alerting: effect of 24 h of prolonged wakefulness. AB - It is well known that a decrease in vigilance can easily occur during 24 h of continuous wakefulness, but no study has assessed whether and in what way extended wakefulness might affect spatial orienting. In other words, it is not clear what happens when a subject has to orient his attention during a state of poor vigilance, resulting from sleep loss or sustained wakefulness. The aim of this study was to investigate this issue. Twelve right-handed male subjects participated in the experiment, which took place on two consecutive days. On the first day, in order to evaluate baseline orienting attention, the subjects performed a covert orienting task (in which the cue stimuli generated endogenous shifts of attention), lasting 20 min; on the second day, during 24 h of prolonged wakefulness, the same task was performed 12 times, about every 120 min, beginning at 10.00 a.m. Results showed an overall slowing of reaction time across the sessions, indicating a linear decrease of vigilance. However, this vigilance decrease did not seem to affect attention-orienting mechanisms, suggesting that the two systems are independent of each other. PMID- 16308690 TI - Electrical activation of the human vestibulo-sympathetic reflex. AB - Muscle sympathetic nerve activity (MSNA) is modulated on a beat-to-beat basis by the baroreflex. Vestibular input from the otolith organs also modulates MSNA, but characteristics of the vestibulo-sympathetic reflex (VSR) are largely unknown. The purpose of this study was to elicit the VSR with electrical stimulation to estimate its latency in generating MSNA. The vestibular nerves of seven subjects were stimulated across the mastoids with short trains of high frequency, constant current pulses. Pulse trains were delivered every fourth heartbeat at delays of 300-700 ms after the R wave of the electrocardiogram. Vestibular nerve stimulation given 500 ms after the R wave significantly increased baroreflex driven MSNA, as well as the diastolic blood pressure threshold at which bursts of MSNA occurred. These changes were specific to beats in which vestibular stimulation was applied. Electrical stimulation across the shoulders provided a control condition. When trans-shoulder trials were subtracted from trials with vestibular nerve stimulation, eliminating the background baroreflex-driven sympathetic activity, there was a sharp increase in MSNA beginning 660 ms after the vestibular nerve stimulus and lasting for about 60 ms. The increase in the MSNA produced by vestibular nerve stimulation, and the associated increase in the diastolic blood pressure threshold at which the baroreflex-driven bursts occurred, provide evidence for the presence of a short-latency VSR in humans that is likely to be important for the maintenance of blood pressure during rapid changes in head and body position with respect to gravity. PMID- 16308691 TI - Planning and drawing complex shapes. AB - Arm and hand movements are generally controlled using a combination of sensory based and memory-based guidance mechanisms. This study examined similarities and differences in visually-guided and memory-guided arm movements, and sought to determine as to what extent certain control principles apply to each type of movement. In particular, the 2/3 power law is a principle that appears to govern the formation of complex, curved hand trajectories; it specifies that the tangential velocity should be proportional to the radius of curvature raised to an exponent of 1/3. A virtual reality system was used to project complex target paths in three-dimensional (3D) space. Human subjects first tracked (with the tip of a handheld pen) a single target moving along an unseen path. The entire target path then became visible and the subject traced the shape. Finally, the target shape disappeared and the subject was to draw it, in the same 3D space, from memory. Most aspects of the movements (speed, path size, shape and arm postures) were very similar across the three conditions. However, subjects adhered to the 2/3 power law most closely in the tracing condition, when the entire target path was visible. Also, only within the tracing condition, there were significant differences in the value of the exponent depending on the size and the spatial orientation of the trajectory. In the tracking and drawing conditions, the exponent was greater than 1/3, indicating that subjects spent more time in areas of tight curvature. This may represent a strategy for learning and remembering the complex shape. PMID- 16308692 TI - T1 and T2 relaxivity of intracellular and extracellular USPIO at 1.5T and 3T clinical MR scanning. AB - In this study we evaluated the effects of intracellular compartmentalization of the ultrasmall superparamagnetic iron oxide (USPIO) ferumoxtran-10 on its proton T1 and T2 relaxivities at 1.5 and 3T. Monocytes were labeled with ferumoxtran-10 by simple incubation. Decreasing quantities of ferumoxtran-10-labeled cells (2.5x10(7)-0.3x10(7) cells/ml) and decreasing concentrations of free ferumoxtran 10 (without cells) in Ficoll solution were evaluated with 1.5 and 3T clinical magnetic resonance (MR) scanners. Pulse sequences comprised axial spin echo (SE) sequences with multiple TRs and fixed TE and SE sequences with fixed TR and increasing TEs. Signal intensity measurements were used to calculate T1 and T2 relaxation times of all samples, assuming a monoexponential signal decay. The iron content in all samples was determined by inductively coupled plasma atomic emission spectrometry and used for calculating relaxivities. Measurements at 1.5T and 3T showed higher T1 and T2 relaxivity values of free extracellular ferumoxtran-10 as opposed to intracellularly compartmentalized ferumoxtran-10, under the evaluated conditions of homogeneously dispersed contrast agents/cells in Ficoll solution and a cell density of up to 2.5x10(7) cells/ml. At 3T, differences in T1-relaxivities between intra- and extracellular USPIO were smaller, while differences in USPIO T2-relaxivities were similar compared with 1.5T. In conclusion, cellular compartmentalization of ferumoxtran-10 changes proton relaxivity. PMID- 16308693 TI - Medium term results in keratoprostheses with biocompatible and biological haptic. AB - PURPOSE: Corneal grafts or limbal stem cell transplantation are often unsuccessful in patients with severe ocular surface disorders such as severe dry eye syndrome, symblepharon or diffuse vascularisation. In those patients, a keratoprosthesis (KPro) may be an alternative for the recovery of vision. Various KPro differ from each other in the material of the haptic that supports the optic cylinder. The haptic may be made of biocompatible or biological material such as tibia bone (TKPro) or dentine and alveolar bone (osteo-odonto-keratoprosthesis, OOKP). On the basis of our experience, we wanted to comment on the value of different KPro. METHODS: Over the last 10 years we have implanted a total of 35 KPro, 29 with biological haptic (25 OOKP and four TKPro), and six KPro with biocompatible haptic (one Legais KPro, five Pintucci KPro). A follow up examination was carried out approximately every6 months. RESULTS: The patients gained a visual acuity of > or =0.9 in 20.6%, of > or =0.5 in 52.9%, of > or =0.2 in 61.8% and a significant improvement in visual acuity in 76.5%, respectively. There was no significant difference between the various types of KPro concerning the best postoperative visual acuity. All patients showing poor improvement had a pre-existing end stage secondary glaucoma or other retinal damage. The median follow-up was 2.9 years (maximum 8) for OOKP, 1 year for TKPro, 1 year (maximum 2) for Pintucci Kpro and 6 month for Legeais KPro. During this period, only one of the KPro with biological haptic was lost (one TKPro after 1 year), compared with four out of six of the KPro with biocompatible haptic (P<0.0001). CONCLUSIONS: Fixation of the KPro by a root of the patient s own tooth (OOKP) leads to the best results in the long-term follow up, as our results as well as the literature demonstrate. As long as a KPro is in place, the visual acuity is as good as the retinal function. For the ranking of different types of KPro, the percentage and the duration of the anatomic success are most important. The comparability of the various KPro results may be limited, since the patients were not randomised and the four groups differ in number. PMID- 16308694 TI - Constancy of organellar genome copy numbers during leaf development and senescence in higher plants. AB - In higher plants, plastid and mitochondrial genomes occur at high copy numbers per cell. Several recent publications have suggested that, in higher plants like Arabidopsis and maize, chloroplast DNA is virtually absent in mature and old leaves. This conclusion was mainly based on DAPI staining of isolated chloroplasts. If correct, the finding that chloroplasts in mature leaves lack DNA would change dramatically our understanding of gene expression, mRNA stability and protein stability in chloroplasts. In view of the wide implications that the disposal of chloroplast DNA during leaf development would have, we have reinvestigated the age dependency of genome copy numbers in chloroplasts and, in addition, tested for possible changes in mitochondrial genome copy number during plant development. Analyzing chloroplast and mitochondrial DNA amounts in Arabidopsis and tobacco plants, we find that organellar genome copy numbers remain remarkably constant during leaf development and are present in essentially unchanged numbers even in the senescing leaves. We conclude that, during leaf development, organellar gene expression in higher plants is not significantly regulated at the level of genome copy number and we discuss possible explanations for the failure to detect DNA in isolated chloroplasts stained with DAPI. PMID- 16308695 TI - [Imaging of cartilage]. AB - The motivation for cartilage repair is the preservation of adequate joint motion. Repairing joint surface congruity and providing balanced load bearing are crucial for this. MRI can contribute to this goal by describing number, depth, size, and distribution of cartilage lesions throughout the different joint compartments. Essential to such a contribution are adequate spatial resolution at a reasonable SNR together with good contrast between both cartilage and the subchondral bone as well as the joint space. For TSE sequences, this is achieved using TEs between 30 and 50 ms. Diagnostic accuracy is optimal when a lesion is depicted in more than one plane. Short TE, high bandwidth, and the appropriate orientation of the frequency encoding direction contribute to minimizing metal artifacts. Besides internal alterations of the cartilage's matrix, moderately T2-weighted TSE sequences sensitively depict bone marrow edema such as signal alterations and joint effusion, both contributing to highlight even subtle cartilage lesions. T1 weighted FS/WE 3D GE sequences profit from their high spatial resolution to appreciate gradual erosion of the cartilage. In OD the interface to the surrounding bone, the integrity of the overlying cartilage, and associated cysts are used to determine stability. The presence of two or more findings increases diagnostic accuracy. Prognosis is associated with the size of the affected area. PMID- 16308696 TI - An amino acid transporter involved in gastric acid secretion. AB - Gastric acid secretion is regulated by a variety of stimuli, in particular histamine and acetyl choline. In addition, dietary factors such as the acute intake of a protein-rich diet and the subsequent increase in serum amino acids can stimulate gastric acid secretion only through partially characterized pathways. Recently, we described in mouse stomach parietal cells the expression of the system L heteromeric amino acid transporter comprised of the LAT2-4F2hc dimer. Here we address the potential role of the system L amino acid transporter in gastric acid secretion by parietal cells in freshly isolated rat gastric glands. RT-PCR, western blotting and immunohistochemistry confirmed the expression of 4F2-LAT2 amino acid transporters in rat parietal cells. In addition, mRNA was detected for the B(0)AT1, ASCT2, and ATB(0+) amino acid transporters. Intracellular pH measurements in parietal cells showed histamine induced and omeprazole-sensitive H+-extrusion which was enhanced by about 50% in the presence of glutamine or cysteine (1 mM), two substrates of system L amino acid transporters. BCH, a non-metabolizable substrate and a competitive inhibitor of system L amino acid transport, abolished the stimulation of acid secretion by glutamine or cysteine suggesting that this stimulation required the uptake of amino acids by system L. In the absence of histamine glutamine also stimulated H+ extrusion, whereas glutamate did not. Also, phenylalanine was effective in stimulating H+/K+-ATPase activity. Glutamine did not increase intracellular Ca2+ levels indicating that it did not act via the recently described amino acid modulated Ca2+-sensing receptor. These data suggest a novel role for heterodimeric amino acid transporters and may elucidate a pathway by which protein-rich diets stimulate gastric acid secretion. PMID- 16308697 TI - Phase I and pharmacokinetic study of edotecarin, a novel topoisomerase I inhibitor, administered once every 3 weeks in patients with solid tumors. AB - PURPOSE: Edotecarin (J-107088) is a potent indolocarbazole topoisomerase I inhibitor which is structurally distinct from the camptothecins. This study aimed to determine the maximum tolerated dose (MTD), the recommended dose for future Phase II studies and the safety, pharmacokinetic profile, and preliminary antitumor activity of edotecarin in a population of patients with advanced solid tumors. EXPERIMENTAL DESIGN: Edotecarin was administered as a single dose by IV infusion over 2 h every 21 days (with 1 week permitted for recovery from toxicities, if needed) in patients with advanced solid tumors. Doses ranged from 8 to 15 mg/m(2). Pharmacokinetic assessments were performed during and after the first administration. RESULTS: Twenty-four patients received 61 cycles of therapy. Dose-limiting toxicities (infection, febrile neutropenia, constipation, ileus, and prolonged grade 4 granulocytopenia) were observed in 3 of 5 evaluable patients at the 15 mg/m(2) dose, defining the MTD. The most commonly reported non hematologic toxicities were anorexia, nausea, malaise, and constipation. Diarrhea was neither frequent nor severe. Neutropenia was the most common hematologic toxicity (grade 3-4 in 21/23 patients during cycle 1). Plasma concentrations of edotecarin rose rapidly following the start of the 2-hour infusion, reaching C (max) values of 103+/-17 ng/ml at the 13 mg/m(2) dose, and decreased steeply after the end of the infusion. Plasma concentrations declined to approximately 1 2 ng/ml at 26 h post start of infusion, the last PK sampling time point. The mean apparent plasma half-life of the drug was 20 h, which should be considered a preliminary estimate until results from studies with a longer duration of plasma sampling are available. A mean of 1.4-3.6% of the dose was recovered as unchanged drug in the urine over 48 h. Unconfirmed tumor regression > or =50% was observed in 2 patients, 1 with metastatic gastric carcinoma and 1 with esophageal cancer. CONCLUSIONS: The MTD of edotecarin administered IV over 2 h every 21 days was 15 mg/m(2). The recommended dose for Phase II studies with a 3-week schedule (with 1 week permitted for recovery from toxicities, if needed) is 13 mg/m(2). The observed safety profile and preliminary evidence of antitumor activity warrant further investigation of this drug in solid tumors. PMID- 16308698 TI - Biweekly regimen of cisplatin, gemcitabine and vinorelbine for advanced non-small cell lung cancer. AB - PURPOSE: Improving chemotherapeutic efficacy in non-small cell lung cancer (NSCLC) will require the development of new strategies to better use currently available agents. To assess the efficacy and safety of a biweekly regimen of cisplatin, gemcitabine and vinorelbine for advanced non-small-cell lung cancer. METHODS: Patients with selected stage IIIb (pleural effusion)/stage IV NSCLC, performance status of 0-2 and normal organ function were eligible. Treatment consisted of cisplatin 100 mg/m(2) on day 1 plus gemcitabine, 1,000 mg/m(2) and vinorelbine 25 mg/m(2) on days 1 and 15 every 28 days. RESULTS: Of the 40 patients enrolled and assessable for response, there were five (12.5%) with confirmed complete response and 14 (35%) with a confirmed partial response for an overall response rate of 47.5%. Nine patients had stable disease while 12 (30%) progressed. Median progression-free survival and overall survival for all patients were 6.3 and 11.1 months, respectively. Toxicity was principally hematologic, with grade 3-4 neutropenia in 30%, and grade 3-4 nausea/vomiting in 22.5%. There were no treatment-related deaths. CONCLUSIONS: The biweekly regimen of cisplatin, gemcitabine and vinorelbine is associated with a high rate of response, lesser toxicity than other three-drug regimens and no benefit of survival. Therefore, the regimen under study may be an appealing alternative when considering other treatment modalities for advanced lung cancer, such as neoadjuvant therapy. PMID- 16308699 TI - CHOP plus etoposide and gemcitabine (CHOP-EG) as front-line chemotherapy for patients with peripheral T cell lymphomas. AB - OBJECTIVE: The present study evaluated the feasibility of CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) plus etoposide and gemcitabine (CHOP EG) as front-line chemotherapy in patients with peripheral T cell lymphomas (PTCLs). PATIENTS AND METHODS: Twenty-six patients with newly diagnosed PTCLs were enrolled into the pilot study. Treatment consisted of classical CHOP plus etoposide 100 mg/m(2) intravenously (i.v.) on day 1 and gemcitabine 600 mg/m(2) i.v. on day 1 in a 3 week interval. RESULTS: Fifteen complete responses (CR, 57.7%) or one unconfirmed complete response (uCR, 3.8%) and four partial responses (PR, 15.4%) were confirmed, giving an overall response rate of 76.9% (95% CI, 58.3-96.3%). Median survival has not yet been reached, while median event free survival was 215 days at a median follow-up duration of 383 days. Estimated overall survival at 1 year was 69.6%. The most severe haematological adverse event was neutropaenia, which occurred with a grade 4 intensity in 14 patients (53.8%). Additionally, febrile neutropaenia was observed in four patients (15.4%). However, there was no treatment-related death. CONCLUSION: The CHOP-EG regimen was found to be feasible in patients with PTCLs. For further investigation on the role of gemcitabine in the treatment of PTCLs, a more large scale phase II or phase III study is warranted. PMID- 16308700 TI - RNA silencing in Chlamydomonas: mechanisms and tools. AB - The generation of a comprehensive EST library and the sequencing of its genome set the stage for reverse genetics approaches in Chlamydomonas reinhardtii. However, these also require tools for the specific downregulation of target gene expression. Consequently, a large number of diverse constructs were developed aimed at reducing target gene expression in Chlamydomonas via the stable expression of antisense or inverted repeat-containing RNA. Double-stranded RNA (dsRNA) generated by the annealing of antisense and sense RNAs or by hairpin formation of an inverted repeat, feeds into the RNA silencing pathway. In this pathway, dsRNA is cleaved into approximately 25-bp small interfering RNAs (siRNAs) by the endonuclease Dicer. One of the two complementary strands of a siRNA is then loaded onto an Argonaute-like protein present as core component within larger complexes. Guided by this single-stranded RNA, the Argonaute-like protein either detects homologous transcripts and cleaves these endonucleolytically, or initiates transcriptional gene silencing. This article summarizes current information derived mainly from the Chlamydomonas genome project on components that are assumed to be involved in RNA silencing mechanisms in Chlamydomonas. Furthermore, all approaches employed in Chlamydomonas to date to downregulate target gene expression by antisense or inverted repeat constructs are reviewed and discussed critically. PMID- 16308701 TI - Study of the autoantibody profile after the acute phase of Kawasaki disease in a cohort of children from North India. AB - A study of organ-specific and organ-nonspecific autoantibodies was carried out in 21 children being followed up after a diagnosis of Kawasaki disease at a tertiary care center of North India. Anti-nuclear antibodies were detected in 9.5% patients while anti-thyroid microsomal antibodies were detected in 23.9% patients. Other autoantibodies (e.g. anti-parietal cell antibody, anti-liver kidney microsomal antibody anti-neutrophil cytoplasmic antibody, anti mitochondrial antibody and anti-smooth muscle antibody) were not detected in any of our patients. Children with Kawasaki disease need to be monitored for the development of autoantibodies during follow-up. PMID- 16308702 TI - Differential expression of manganese superoxide dismutase sequence variants in near isogenic lines of wheat during cold acclimation. AB - Numerous sequence variants of wheat (Triticum aestivum L.) manganese superoxide dismutase (MnSOD) genes have been found. Quantitative real-time PCR was used to measure the expression levels of three MnSOD genes distinguished by a variable amino acid, and three genes distinguished by sequence variation in the 3' untranslated region (3' UTR), in wheat plants grown at 20 degrees C and cold acclimated for 1-4 weeks at 2 degrees C. The amino acid variants did not differ significantly in expression levels, however, differential expression of genes differing in the 3' UTR was observed. Diploid wheat-related species also carried sequence variants of MnSOD, with differing levels of expression, suggesting diversification of the MnSOD gene family occurred prior to the polyploidization events of hexaploid wheat. PMID- 16308703 TI - Antennal lobe projection destinations of Helicoverpa zea male olfactory receptor neurons responsive to heliothine sex pheromone components. AB - We used single sensillum recordings to define male Helicoverpa zea olfactory receptor neuron physiology followed by cobalt staining to trace the axons to destination glomeruli of the antennal lobe. Receptor neurons in type A sensilla that respond to the major pheromone component, (Z)-11-hexadecenal, projected axons to the cumulus of the macroglomerular complex (MGC). In approximately 40% of these sensilla a second receptor neuron was stained that projected consistently to a specific glomerulus residing in a previously unrecognized glomerular complex with six other glomeruli stationed immediately posterior to the MGC. Cobalt staining corroborated by calcium imaging showed that receptor neurons in type C sensilla sensitive to (Z)-9-hexadecenal projected to the dorsomedial posterior glomerulus of the MGC, whereas the co-compartmentalized antagonist-sensitive neurons projected to the dorsomedial anterior glomerulus. We also discovered that the olfactory receptor neurons in type B sensilla exhibit the same axonal projections as those in type C sensilla. Thus, it seems that type B sensilla are anatomically type C with regard to the projection destinations of the two receptor neurons, but physiologically one of the receptor neurons is now unresponsive to everything except (Z)-9-tetradecenal, and the other responds to none of the pheromone-related odorants tested. PMID- 16308704 TI - Clinical characteristics and surgical treatment of perianal and perineal rhabdomyosarcoma: analysis of Japanese patients and comparison with IRSG reports. AB - Rhabdomyosarcomas of the perianal and perineal regions are uncommon. This study was performed to clarify the clinical characteristics and guidelines of surgical treatment of patients with perianal and perineal rhabdomyosarcomas younger than 20 years of age. Twenty-nine patients, 26 patients identified in the Japanese literature and three of our own, were analyzed and the results were compared with the data reported from the Intergroup Rhabdomyosarcoma Study Group (IRSG). Female predominance and a twin-peak age distribution in infancy and adolescence were characteristic findings of the Japanese patients that were not observed in the IRSG studies. The demographic differences between the two groups were attributed to the differences in demographics of patients younger than 10 years of age. Of the 29 patients, 17 were categorized into clinical groups III/IV and 21 patients into stages 3/4. Alveolar histology was diagnosed in 18 patients. In patients more than 10 years of age, the female predominance was more prominent and the incidences of advanced clinical groups/stages and alveolar histology were significantly higher than those in patients younger than 10 years of age. Inguinal lymph nodes were always involved in patients with lymph node metastases and three patients developed metastases to the breast. Information regarding the survival time was available for 18 patients and the 5-year overall survival was 20%. Two patients with a group I/stage 2 tumor and one with a group II/stage 3 tumor survived for more than 2 years with no evidence of the disease. In these patients, the tumors were excised by primary surgery or primary reexcision and they were not accompanied by lymph node metastasis. Based on these data, complete tumor resection prior to chemotherapy should be pursued and the inguinal lymph nodes should be at least sampled because nodal involvement is closely associated with the patient's prognosis. PMID- 16308705 TI - The use of refluxing ureter in the creation of a Mitrofanoff channel in children undergoing bladder augmentation: is a formal reimplantation necessary? AB - To study the fate of a refluxing ureteral stump used as a Mitrofanoff channel (MC) for clean intermittent catheterization (CIC), without reimplantation in augmented bladders. Ten boys and seven girls, 1-14 years old (mean 6.5 years) seen in period 1998-2005, underwent creation of MC using refluxing or potentially refluxing ureters. Concomitant bladder augmentation (BA) was performed for various abnormalities of lower urinary tract, including exstrophy epispadias complex, neurogenic bladder cloaca and posterior urethral valve. The uretero vesical junction was maintained intact during creation of the MC. Follow-up period ranged from 2-72 months (mean 32.5 months). Satisfactory CIC was possible in 14 children while in two children the stoma became obstructed due to non usage. Urinary leak via the ureteric MC was seen in only one patient who partially responded to anticholinergic therapy and a shortened CIC interval but this needed to be formally closed subsequently. A refluxing ureter can be used in the creation of a MC without formal reimplantation in the majority of children undergoing BA. PMID- 16308706 TI - Ill health retirement in Scottish teachers: process, outcomes and re-employment. AB - OBJECTIVES: To assess the process, causes and outcomes of ill health retirement (IHR) in teachers in Scotland. Perceptions and experience of occupational health services (OHS), access to rehabilitation and redeployment, current health, post retirement experience and predictors of re-employment were identified. METHODS: 537 teachers who retired due to ill health between April 1998 and March 2000 were mailed our IHR questionnaire by the Scottish Public Pensions Agency. RESULTS: The most common cause of IHR was mental disorders (37%), followed by diseases of the musculoskeletal system (18%). 11% of teachers attended an OHS prior to IHR. 9% of teachers were offered part-time work and 5% were offered alternative work. 63% of retired teachers stated their health had improved and 48% said they would like to work again. 36% of the surveyed teachers have found re-employment since their retirement. On unadjusted analyses, re-employment of teachers after IHR was significantly associated with sex, having dependants, job group, cause of IHR, health having improved and wanting to work again. Multiple logistic regression analyses showed three variables as independent predictors of re-employment: having dependants, job group and cause of IHR. CONCLUSION: This is the first comprehensive study investigating teachers' experiences of IHR in Scotland. The findings highlight substantial lack of support for teachers in a number of areas which need to be addressed by managers and employers. Our study confirms the need for rehabilitation and teacher support services in order to try and retain experienced teachers in the profession. PMID- 16308707 TI - Control by cytokinins of the cellular behavior in the plate meristem of zucchini cotyledons. AB - The temporal and spatial effects of exogenous cytokinins on both cell expansion and division activity in the plate meristem of cultured zucchini cotyledons were studied. N6-benzylaminopurine (1-100 microM) and N-(2-chloro-4pyridyl)-N' phenylurea (4PU-30) (0.1-100 microM) greatly stimulated the cell growth and division. They provoked multiple cell cycles, formation of larger clusters of daughter cells and an increase of the final number of cells. Both cytokinins led to earlier achievement of final cotyledon size and shortened the cell doubling time. By contrast to the purine cytokinin, phenylurea cytokinin 4PU-30 enlarged the cotyledon predominantly in length. Zeatin and kinetin were less effective, particularly in stimulating cell expansion. In low concentrations, all cytokinins were more effective in stimulating division activity rather than expansion. The cells in the cotyledon margins displayed a higher division activity, especially when treated with exogenous cytokinins. The final cotyledon and cluster areas were not of the strict proportional dependence upon the number of their cells. These results provide a novel example where stimulated cell division fails to evoke a respective increase in the final organ size. PMID- 16308708 TI - Sporadic Cajal cell hyperplasia is common in resection specimens for distal oesophageal carcinoma. A retrospective review of 77 consecutive surgical resection specimens. AB - Interstitial cell of Cajal (ICC) hyperplasia has been documented in conditions associated with multiple gastrointestinal stromal tumours (GISTs) (familial GIST syndromes, Carney's triad and von Recklinghausen's disease) and rarely in the vicinity of sporadic GISTs. The incidence of sporadic ICC hyperplasia and the so called seedling leiomyoma (SLM) of the lower oesophagus has not been studied in the KIT era. In a retrospective review of 77 consecutive, routinely processed oesophagogastric resection specimens for distal oesophageal carcinoma, we found foci of ICC hyperplasia in 7 of 77 (9.1%) cases and foci of SLM in 17 of 77 (22%) cases. Two types of ICC hyperplasia were recognized: a non-circumscribed type and a nodular expansile type with peripherally compressed myenteric neural tissues. All cases of ICC hyperplasia were vimentin+/CD34+/CD117+. SLMs were desmin+/vimentin(-)/CD34(-)/CD117(-), similar to smooth muscles of the gut wall. In a prospective study of 32 non-carcinomatous specimens from age-matched patients (mostly autopsy cases), we found SLMs in only one case, but we were unable to detect ICC hyperplasia in any of the cases. We concluded that sporadic KIT-positive spindle-cell hyperplasia and SLMs were unexpectedly common in distal oesophageal specimens harbouring carcinomas. The possible mechanisms leading to the development of these putative precursor lesions will be discussed. PMID- 16308709 TI - Second-line strategies for metastatic renal cell carcinoma: classics and novel approaches. AB - OBJECTIVES: Renal cell carcinoma is an aggressive malignancy with a high propensity for both early and metachronous regional and distant metastasis. While surgical resection is the mainstay of therapy for patients with localized disease, the prognosis for patients with distant metastasis is poor with a 5-year survival rate of less than 10%. Response rates to first-line immunotherapy or immunochemotherapy range from 10-35%; responses achieved are predominantly partial remissions of short duration. Until today, there is no standard therapeutic procedure for the growing number of patients who relapse following first-line therapy and desire further active treatment. MATERIALS AND METHODS: This article reviews classic and recent publications about second- and third-line approaches, their potential efficacy and toxicity. RESULTS: Several novel approaches have raised well-founded hope. Especially the application of monoclonal antibodies targeting VEGF signalling as well as different receptor tyrosine kinase inhibitors have the potential to change the face of second-line treatment of patients with metastatic RCC. Both groups of agents are focused in current phase III trials, either as mono- and/or combination therapy. CONCLUSIONS: Until today, second-line treatment of patients with metastatic RCC progressing under therapy with biological response modifiers remains an unresolved issue. The results of ongoing clinical trials evaluating novel targeted approaches can be expected with suspense. PMID- 16308710 TI - [New perspectives for simulator-based training in paediatric anaesthesia and emergency medicine]. AB - Anaesthesia and emergency medical care for infants and toddlers is often associated with high clinical demands and specific challenges. Nevertheless, a significant proportion of interventions is performed by anaesthetists and emergency physicians with no specialised paediatric training and little experience in the management of anaesthetic incidents and emergencies specific to these age groups. Extensive studies have demonstrated a close inverse correlation between the level of specialisation and perioperative morbidity and mortality. However, clinical circumstances and the relatively small number of paediatric cases at many institutions often hinder the establishment of improved training concepts. Recently, high-fidelity infant simulators have become available, which permit authentic exposure to a large spectrum of scenarios in paediatric anaesthesia and emergency medicine. A multimodular concept of training, including such simulator-based techniques, may relieve the widespread shortage in clinical experience, and hence greatly facilitate improvement of quality of care and patient safety. PMID- 16308711 TI - [Diagnostics and treatment of prostate cancer after kidney transplantation]. AB - The number of patients with prostate cancer and end-stage renal disease or prostate cancer following kidney transplantation has continuously increased in industrialized countries. According to the data generated by Penn et al. a higher incidence of prostate cancer following kidney transplantation can be seen but is probably due to a more intense screening of the recipients. It is rather a common opinion that no elevated risk of prostate cancer following kidney transplantation exists. In patients with strictly localized prostate cancer curative treatment should be the aim also after kidney transplantation. Kidney transplantation does not interfere with surgical access to the prostate gland (retropubic or perineal). Nonlocal tumors of the prostate should also be treated following the general recommendations regarding prostate cancer. Looking at the current literature, a reduction or change of immunosuppression seems reasonable. It is necessary to establish a nationwide (or even European) cancer register, especially for patients before and after transplantation. PMID- 16308712 TI - [The male genital in legal medicine]. AB - Pathomorphological findings for the male genital are an exception among forensic cases. In earlier publications they were usually published as individual cases. A significant percentage are the so-called fertility- and potency medical certificates, which are usually commissioned for doubtful paternity cases. Old forensic writings about genital injuries distinguish--apart from the necessity of a complete dissection--between "deadly as such" and "accidentally deadly" injuries. Another group consists of reports about self-inflicted genital mutilation, e.g. among prison inmates or soldiers of the last world wars to become an invalid. When it was necessary to identify a corpse, e.g. in cases of progressive decomposition, Merkel recommended in 1945 the representation of the so-called "Schnepfenkopf" (snipe head; colliculus seminalis) in the lesser pelvis. In this article a medical and forensic review of forensically relevant findings of the male genital is presented and illustrated with historical cases. PMID- 16308713 TI - Predictive value of electrocorticography in epilepsy patients with unilateral hippocampal sclerosis undergoing selective amygdalohippocampectomy. AB - The purpose of this study was to evaluate the predictive value of intraoperative electro-corticography (ECoG) in patients with unilateral hippocampal sclerosis (HS) undergoing transsylvian selective amygdalohippocampectomy (sAHE). ECoG was recorded before and after resection in 22 patients with medication-resistant mesial temporal lobe epilepsy. The sAHE was performed, regardless of ECoG findings. ECoG findings recorded from the mesiobasal temporal lobe (MTL) and lateral temporal lobe (LTL) before and after the sAHE were correlated with seizure outcome 12 months later. Ten patients had right-sided and 12 left-sided HS. Average age was 37.1 years. Pre-resection spikes were restricted to the MTL in 11 patients and to the LTL in one. In three patients spikes were recorded from MTL and LTL and in seven no spikes were recorded before the resection. Fifteen patients (68%) remained completely seizure-free and 19 (86%) were in Engel's class I post-operatively. Patients with pre-resection spikes restricted to the MTL (n = 11) remained seizure-free more frequently (9/11, 82%) compared with other patients (6/11, 55%; P = 0.36). Pre-resection ECoG may be helpful in the prediction of seizure outcome in patients undergoing sAHE for mesial temporal lobe epilepsy. A larger study including more than 100 patients is needed to determine the predictive value of ECoG in patients with mesial temporal lobe epilepsy. PMID- 16308714 TI - Proteome analysis and comparison of Clostridium acetobutylicum ATCC 824 and Spo0A strain variants. AB - The proteomic profiles of several Clostridium acetobutylicum strains were compared by two-dimensional gel electrophoresis and mass spectroscopy. The proteomic profile of C. acetobutylicum wild type strain ATCC 824 with and without a commonly used control plasmid and with a spo0A overexpression plasmid pMSPOA was compared. A total of 2,081 protein spots were analyzed; 23 proteins were chosen to be identified of which 18 were unique and 5 were proteins located in more than one location. The proteins identified were classified into heat shock stress response, acid and solvent formation, and transcription and translation proteins. Spo0A was identified and its protein expression was confirmed to be absent in the spo0A knockout SKO1 strain as expected, as was the protein Adc, which is known to be regulated by Spo0A. The expression of six proteins was not detected in strain SKO1 indicating these proteins require Spo0A. Spo0A overexpression affected the abundance of proteins involved in glycolysis, translation, heat shock stress response, and energy production. Two features were identified: five of the 23 proteins identified were located in more than one position and clusters of protein spots resembled fingers of a straightened hand. Normally a protein localizes to only one spot on the gel; localization of a protein to more than one spot is indicative of post-translational modifications, suggesting that such modification of proteins may be a more prevalent mechanism in C. acetobutylicum than previously thought. The clusters of protein spots resembling fingers of a straightened hand were in the acidic high molecular weight areas. Two such protein spots were identified as variants of the same protein, GroEL. PMID- 16308715 TI - Marrow changes in anorexia nervosa masking the presence of stress fractures on MR imaging. AB - Patients with anorexia nervosa (AN) usually have abnormal bone and bone marrow metabolism resulting in osteopenia and serous bone marrow change. There is an increased risk of stress/insufficiency fractures and these can be the first presentation of AN. This case report describes a patient with previously undiagnosed AN who presented with foot pain. The serous bone marrow changes of AN were found to mask the MR imaging features of stress fractures, as both had low T1w and high T2w and STIR signal intensities. Contrast enhancement was not helpful but actually masked fractures. Scintigraphy was helpful. The radiologist might be the first clinician to raise the possibility of AN and should be aware of the difficulties in diagnosing stress fractures in bones with underlying serous bone marrow change. In this severe case of AN even the heel fat pad and the fat pad in Kager's triangle had undergone serous change. PMID- 16308716 TI - Endometriosis of the vastus lateralis muscle. AB - We report an unusual case of a woman who sought medical attention for a painful mass within her anterolateral left thigh. The patient's symptoms waxed and waned with her menses. Magnetic resonance imaging revealed a 3 cm mass isointense to muscle and surrounded by a rim of decreased signal intensity on T1-weighted images. T2-weighted images with fat saturation revealed a low signal intensity mass with a surrounding rind of high signal intensity edema. Based on its magnetic resonance imaging characteristics, the lesion was initially thought to represent the sequela of prior trauma or perhaps a small fibrous tumor. An excisional biopsy of the mass, however, revealed endometrial tissue, and a pathologic diagnosis of intramuscular endometriosis was rendered. PMID- 16308717 TI - Magnetic resonance imaging of bilateral lateral congenital dislocations of unossified patellae. AB - We describe our experience using magnetic resonance imaging (MRI) to evaluate the unossified dislocated cartilaginous patella in a 6-year-old male patient with restricted extension and flexion deformity of both knees. MRI is used widely in the visualization of cartilage since it can show cartilage directly. In addition, FS FLASH 3D sequence has recently been accepted as a suitable sequence in the evaluation of hyaline cartilage. MRI makes it possible to evaluate ligamentous, tendinous, muscular, and cartilaginous structures as well as the abnormalities related to them. We applied this technique in our case and found it very effective in locating the unossified dislocated cartilaginous patella. We also observed structural changes such as bilateral lateral displacement of short quadriceps tendon inserting into diminutive patella, insertion of bilateral patellar tendons into anterolateral tibia, and a stretching of the medial collateral ligament associated with valgus stress. PMID- 16308718 TI - Hyperplastic callus formation in osteogenesis imperfecta type V mimicking osteosarcoma: 4-year follow-up with resolution. AB - We report a case of hyperplastic callus formation that occurred in both femurs in a patient with type V osteogenesis imperfecta (OI), with 4-year follow-up and resolution. The clinical, histological and imaging aspects of this condition are discussed. Recognition of the hyperplastic callus formation in this particular type of OI is important in order to avoid misdiagnosis. PMID- 16308719 TI - MRI findings in Little Leaguer's shoulder. AB - Little leaguer's shoulder, a stress injury of the proximal humeral physis, should be considered in the differential diagnosis for an adolescent baseball player with shoulder pain, especially if the player is pitching regularly in a competitive environment. While roentgenographs may or may not be helpful, depending on the duration and severity of the injury, we report the MRI appearance of a case of little leaguer's shoulder. We found MRI helpful in diagnosing injury to the growth plate that was radiographically occult; furthermore, we were able to document the patient's progress with a follow-up MRI examination, which showed improvement with treatment. PMID- 16308720 TI - Decompressive surgery for degenerative lumbar spinal stenosis: long-term results. AB - We report the outcome of 50 patients with degenerative lumbar spinal stenosis who were treated surgically by spinal decompression between 1984 and 1995. Their mean age at the time of surgery was 59.9 (45-77) years and the mean follow-up was 11.6 (6.1-17.2) years. Five patients had a concomitant spinal fusion. The preoperative data were collected retrospectively from the patients' charts. The follow-up data were obtained from a clinical examination and questionnaire including overall pain, ability to work, walking ability, use of analgesics and satisfaction with surgery. The outcome was rated as excellent in 23 patients, good in 13 patients, fair in 9 patients and poor in 5 patients. Patients with concomitant fusion had good to excellent results and were more satisfied, whereas patients with long standing preoperative symptoms had poor to fair result and were less satisfied. PMID- 16308721 TI - The hematopathological basis for studying effects of the demethylating agent 5 aza-2'-deoxycytidine (decitabine) in myelodysplasia. AB - The myelodysplastic syndromes have, since their first recognition decades ago, been considered notoriously difficult with regard to their proper classification, determination of prognosis, and optimal treatment. With the advent of the French American-British (FAB) classification, now aided but not superseded by the World Health Organization classification, distinct biological entities have been delineated, which in turn are very useful for stratification to different, established and experimental treatment modalities. However, precise subclassification of different types of myelodysplastic syndrome (MDS) is only possible with hematopathological studies based on the analysis of peripheral blood, bone marrow smear, and bone marrow biopsy, backed by appropriate clinical information. Bone marrow cytogenetics are also essential for any risk stratification since they still provide the second most powerful prognostic parameter after bone marrow blast enumeration. This paper will review the most important aspects of hematopathological diagnostics in MDS, risk scoring, and their application to the inclusion and stratification of patients into the European Organization for Research and Treatment of Cancer (EORTC)/German MDS Study Group Phase III multicenter trial of low-dose decitabine in patients more than 60 years old with high-risk MDS. Emphasis is placed on itemizing the broad spectrum of cytologic and histologic stigmata defining the myelodysplastic categories that are to be considered in this study. PMID- 16308722 TI - Larval bullfrog skin expresses ENaC despite having no amiloride-blockable transepithelial Na+ transport. AB - Amiloride-blockable Na(+) transport, measured as an amiloride-blockable short circuit current (Am-SCC), is mediated by the epithelial Na(+) channel (ENaC). Am SCC is not normally present in bullfrog tadpole skin, but when such skin is cultured with corticoids an amiloride-blockable Na transport appears. Prolactin (PRL) inhibits its corticoid-induced development. Using specific PCR primers for adult frog ENaC and RT-PCR, we investigated whether corticoids can induce all three ENaC subunits, and whether this expression of ENaC subunit(s) can be blocked by adding PRL with the corticoids. We found that (1) the sequences of the RT-PCR products obtained using primers for alpha-ENaC were identical between larval and adult skins, (2) the mRNAs for all three ENaC subunits were expressed in larval skin under normal conditions despite no amiloride-blockable Na(+) transport being detectable, (3) all three subunits were expressed in larval skins whether they were cultured with corticoids (amiloride-blockable Na transport present) or with corticoids supplemented with PRL (no amiloride-blockable Na transport present). An antibody against a peptide from the alpha-ENaC of adult bullfrog was localized to the apical cells of both larval and adult skins. Since no amiloride-blockable Na transport exists across larval skin under these conditions, these results suggest that ENaC protein was expressed prior to the onset of transport. ENaC may be in the plasma membrane in an inactivated form or, alternatively, within vesicles waiting to be inserted. PMID- 16308723 TI - Pre-and postoperative psychological characteristics in mothers of patients with idiopathic scoliosis. AB - The purpose of the present study was to clarify changes in the psychological state of mothers of patients with idiopathic scoliosis, and to clarify relationships between the psychological states of the mothers and patients. The Maudsley personality inventory (MPI) was administered to 30 patients with idiopathic scoliosis who underwent surgery and their mothers preoperatively and at about 1 year postoperatively. We investigated the relationships between preoperative MPI scores and postoperative scores in patients and their mothers, respectively, and the relationships between MPI scores for patients and mothers. The results of the present study revealed that patients became more extroverted following surgery, while mothers displayed reduced neurotic tendencies. In conclusion, the mothers of patients who undergo corrective treatment for scoliosis may have experienced a substantial psychological burden before surgery, and doctors who treat patients with scoliosis should bear this point in mind. PMID- 16308724 TI - Outcome in adolescent idiopathic scoliosis after brace treatment and surgery assessed by means of the Scoliosis Research Society Instrument 24. AB - A retrospectively designed long-term follow-up study of adolescent idiopathic scoliosis (AIS) patients who had completed treatment, of at least 2 years, by means of brace, surgery, or both brace and surgery. This study is to assess the outcome after treatment for AIS by means of the Scoliosis Research Society Outcome Instrument 24 (SRS 24). One hundred and eighteen AIS patients (99 females and 19 males), treated at the Aarhus University Hospital from January 1, 1987 to December 31, 1997, were investigated with at least 2 years follow-up at the time of receiving a posted self-administered questionnaire. Forty-four patients were treated with Boston brace (B) only, 41 patients had surgery (S), and 33 patients were treated both with brace and surgery (BS). The Cobb angles of the three treatment groups did not differ significantly after completed treatment. The outcome in terms of the total SRS 24 score was not significantly different among the three groups. B patients had a significantly better general (not treatment related) self-image and higher general activity level than the total group of surgically treated patients, while surgically treated patients scored significantly better in post-treatment self-image and satisfaction. Comparing B with BS we found a significantly higher general activity level in B patients, while the BS group had significantly higher satisfaction. There were no significant differences between BS and S patients in any of the domain scores. All treatment groups scored "fair or better" in all domain scores of the SRS 24 questionnaire, except in post-treatment function, where all groups scored worse than "fair". Improvement of appearance by means of surgical correction increases mean scores for post-treatment self-image and post-treatment satisfaction. Double treatment by brace and surgery does not appear to jeopardize a good final outcome. PMID- 16308726 TI - Why patients with Alzheimer's disease may show increased sensitivity to tropicamide eye drops: role of locus coeruleus. AB - RATIONALE: Patients suffering from Alzheimer's disease (AD) may show increased sensitivity to tropicamide, a muscarinic cholinoceptor antagonist. AD is associated with a severe loss of noradrenergic neurones in the locus coeruleus (LC), which can be "switched off" experimentally by the alpha(2)-adrenoceptor agonist clonidine. The possibility arises that increased pupillary sensitivity to tropicamide in AD may be due to diminished LC activity. OBJECTIVE: To examine the hypothesis that clonidine may potentiate tropicamide-evoked mydriasis. MATERIALS AND METHODS: Sixteen healthy male volunteers participated in two experimental sessions (0.2 mg clonidine or placebo) conducted 1 week apart. In each session tropicamide (0.01% 10 microl x 2) was applied to the left eye and artificial tear (10 microl x 2) was applied to the right eye. Pupillary functions (resting pupil diameter and light and darkness reflexes), alertness and non-pupillary autonomic functions (blood pressure, heart rate, core temperature and salivary output) were measured. Data were analysed by ANOVA, with multiple comparisons. RESULTS: Tropicamide increased resting pupil diameter, velocity and amplitude of the darkness reflex response, and decreased recovery time of the light reflex response. Clonidine affected all these pupillary measures in the opposite direction with the exception of the recovery time. The mydriatic response to tropicamide was potentiated by pre-treatment with clonidine. Clonidine reduced critical flicker fusion frequency, subjective alertness, blood pressure, salivation and temperature. CONCLUSIONS: The potentiation of tropicamide-evoked pupil dilatation by clonidine may be due to the abolition of the increase in parasympathetically mediated pupil constriction due to reduced LC activity. PMID- 16308727 TI - Assessing DSM-IV nicotine withdrawal symptoms: a comparison and evaluation of five different scales. AB - OBJECTIVE: This study evaluated four of the major scales used to measure nicotine withdrawal symptoms plus one new scale. METHODS: Eighty-three smokers were randomly assigned to continue smoking (n=37) or abstain completely for 24 h (n=46), by which time the symptoms should become manifest. Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) withdrawal symptoms (irritability, depression, restlessness, insomnia, anxiety, hunger and poor concentration) plus craving were measured at baseline and after 24 h. The scales tested were the Minnesota Nicotine Withdrawal Scale (MNWS), the Mood and Physical Symptoms Scale (MPSS), the Shiffman Scale (SS), the Wisconsin Smoking Withdrawal Scale (WSWS) and the newly developed Cigarette Withdrawal Scale (CWS). RESULTS: Measurement of withdrawal symptoms was robust in the case of all scales for total withdrawal score, irritability, restlessness, poor concentration and craving. The MNWS and CWS were less sensitive to depression; the WSWS and MNWS were less sensitive to insomnia; the MPSS was less sensitive to anxiety and hunger; the CWS and WSWS did not include restlessness as a distinct symptom; the SS did not include insomnia, and its scores tended to decline over time during ad lib smoking. Longer scales, using multiple items to measure each symptom, did not yield more reliable or accurate measurement than briefer scales. CONCLUSIONS: To measure total withdrawal discomfort or craving, all of the scales examined can be recommended, and there is little to choose between them apart from length. When it comes to assessing individual symptoms, different scales have different strengths and weaknesses. There would be merits in developing a new questionnaire that combined the best features of the scales tested. PMID- 16308728 TI - A retrospective study on the metacercarial production of Fasciola hepatica from experimentally infected Galba truncatula in central France. AB - A retrospective study of the experiments performed during the past 15 years on infections of Galba truncatula with Fasciola hepatica was carried out to determine what susceptible populations of snails might be used for the commercial production of metacercariae, and to examine this metacercarial production in relation to the characteristics of snail infections. Of the four groups of snail populations studied, the ablest snails to sustain a complete larval development of F. hepatica originated from populations living on siliceous soils at 600 m and more in altitude. In contrast, snail populations living along river banks on siliceous soils were inappropriate due to the poor characteristics of snail infections (high snail mortality, low prevalence of snail infections, and low number of cercariae produced). Except for these last populations, 86-87% of cercaria-shedding (CS) snails in the other populations shed less than 300 cercariae, even if a maximum of 1,772 cercariae were obtained from a single snail. The date of the first cercarial shedding at 20 degrees C began during week 7 or 8 PE for 80.1-83.5% of CS snails. Most metacercariae (82.0-85.9% of the total production) were recorded during the first 10 days of the patent period. In these conditions, the authors collected metacercarial production up to the beginning of week 10 PE (20 degrees C) and did not use snails that shed their cercariae during the following weeks due to too low numbers of parasites. This method allows to have a continuous production of metacercariae over time by using successive groups of infected snails, each being separated from the other by a fortnight's time. PMID- 16308729 TI - Ubiquitination of plasma membrane ectophosphatase in bloodstream forms of Trypanosoma brucei. AB - Bloodstream forms of Trypanosoma brucei contain plasma-membrane-integral acidic ectophosphatase. Here, it is shown by N-terminal sequencing that the ectophosphatase found in ricin-binding material was modified by ubiquitin. Three different ubiquitinated species corresponding to single, double and triple ubiquitinated forms of the enzyme were identified. Immunofluorescence studies with live bloodstream-form parasites showed that the ectophosphatase was localized in the flagellar pocket-the sole site for endocytosis in trypanosomes. As ubiquitin modification of plasma membrane proteins serves as an internalization signal, it is suggested that ubiquitinated ectophosphatase is labelled for endocytosis. PMID- 16308730 TI - Inflammation and glandular duct dilatation of the tongue from patients with chronic Chagas disease. AB - The purpose of this study was to evaluate morphologically the tongue of individuals with chronic Chagas disease (CD) in comparison to the non-chagasic ones. Twenty-four protocol cases of autopsies were selected. They were subdivided into CD patients (10 cases) and non-chagasic ones (14 cases). The morphometric analysis was accomplished for the tongue muscle and salivary glands duct lumen area. In three CD patients, perineuritis was found, and two of them showed megaesophagus and megacolon. The intensity of the inflammation in the von Ebner's glands, the tongue muscles, and the salivary glands duct lumen area was significantly higher in the CD patients. We concluded that the CD patients show salivary glands duct dilatation, which probably would have a relation with alterations in the autonomic nervous system. The inflammation found in CD patients is in accordance with that described in comparative studies on the digestive tract and heart. These morphological findings suggest that the histopathological analysis of the tongue associated with other organs, or even in an isolated manner, can add in the diagnosis and pathogenesis of the CD chronic phase. PMID- 16308731 TI - The inhibition of angiogenesis by antisense oligonucleotides to clusterin. AB - Angiogenesis is a primary disease target in ocular retinopathy and a secondary target in numerous other angiogenic diseases such as cancer, rheumatoid arthritis and psoriasis. Clinical trials using antiangiogenic antisense oligonucleotides (aso's) for the treatment of ocular disorders or cancer are well advanced. Clusterin aso's are currently under investigation for the treatment of prostate cancer. We have investigated the antiangiogenic properties of clusterin aso's using a capillary cell (HUVEC) viability assay. In this study we included aso's to known apoptosis modulators (bcl-2, bcl-xl and survivin) which were previously identified in HUVEC's. We have also studied the effect of clusterin aso's on angiogenesis using an in vitro, matrigel assay and on HUVEC apoptosis using an ELISA DNA fragmentation assay. Clusterin, bcl-2, bcl-xl and survivin aso's were all found to inhibit HUVEC growth. The apoptosis-inducing drugs paclitaxel, camptothecin and doxorubicin were also found to inhibit HUVEC proliferation. Combinations of aso's with these drugs demonstrated a minor additive but not synergistic inhibitory effect on HUVEC proliferation. Clusterin aso's were found to strongly inhibit angiogenesis and induce high levels of apoptosis in HUVECs. In cancer cells the prosurvival protein clusterin may protect the cells from apoptosis-inducing agents so that the clusterin aso's may act as chemosensitization agents. These data demonstrate a strong antiangiogenic action of clusterin aso's, that is not necessarily related to any chemosensitization effect of this agent. PMID- 16308732 TI - EDB fibronectin and angiogenesis -- a novel mechanistic pathway. AB - Extra domain-B containing fibronectin (EDB(+) FN), a recently proposed marker of angiogenesis, has been shown to be expressed in a number of human cancers and in ocular neovascularization in patients with proliferative diabetic retinopathy. To gain molecular understanding of the functional significance of EDB(+) FN, we have investigated possible regulatory mechanisms of induction and its role in endothelial cell proliferation and angiogenesis. Human vascular endothelial cells were cultured in high levels of glucose, and fibrogenic growth factors, transforming growth factor-beta1 (TGF-beta1) and endothelin-1 (ET-1). Our results show that high glucose levels, TGF-beta1, and ET-1 upregulated EDB(+) FN expression. Treatment of cells exposed to high glucose with TGF-beta1 neutralizing antibody and ET receptor antagonist prevented high glucose-induced EDB(+) FN expression. In order to elucidate the functional significance of EDB(+) FN upregulation, cells were subjected to in vitro proliferation and angiogenesis assays following EDB peptide treatment and specific EDB(+) FN gene silencing. Our results show that exposure of cells to EDB peptide increased vascular endothelial growth factor (VEGF) expression, endothelial proliferation, and tube formation. Furthermore, specific EDB(+) FN gene silencing prevented both basal and high glucose-induced VEGF expression and reduced the proliferative capacity of endothelial cells. In conclusion, these results indicate that EDB(+) FN is involved in endothelial cell proliferation and vascular morphogenesis, findings which may provide novel avenues for the development of anti-angiogenic therapies. PMID- 16308734 TI - Formation of the coronary vasculature during development. AB - The formation of the coronary vasculature involves a series of carefully regulated temporal events that include vasculogenesis, angiogenesis, arteriogenesis and remodeling. This review explores these events, which begin with the migration of proepicardial cells to form the epicardium and end with postnatal growth and remodeling. Coronary endothelial, smooth muscle and fibroblast cells differentiate via epithelial-mesenchymal transformation; these cells delaminate from the epicardium. Following the formation of a tubular network by endothelial cells, an aortic ring of endothelial cells penetrates the aorta at the left and right aortic cusps to form the two ostia. Smooth muscle cell recruitment occurs rapidly and the coronary artery network begins forming as blood flow is established. Recent studies have identified a number of regulatory molecules that play key roles in epicardial formation and the transformation of its component cells into mesenchyme. Moreover, we are finally gaining some understanding regarding the interplay of angiogenic growth factors in the complex process of establishing the coronary vascular tree. Understanding coronary embryogenesis is important for interventions regarding adult cardiovascular diseases as well as those necessary to correct congenital defects. PMID- 16308735 TI - Angiogenesis in ischaemic and hypertrophic hearts induced by long-term bradycardia. AB - Angiogenesis and improved left ventricular function as a consequence of long-term bradycardia were first demonstrated in normal hearts, either electrically paced (rabbits, pigs) or treated with a selective sinus blocking drug alinidine (rats). Here we review the evidence that chronic heart rate reduction can have similar effects in the heart with compromised vascular supply, due to either hypertensive or haemodynamic overload hypertrophy (rats, rabbits) or ischaemic damage (rats, rabbits, pigs). Bradycardia induced over several weeks increased capillarity in all hypertrophied hearts, and in border and remote left ventricular myocardium of infarcted hearts. In some, but not all cases, coronary blood flow was improved by heart rate reduction, suggesting enlargement of the resistance vasculature in some circumstances. Cardiac or left ventricular function indices, which were depressed by hypertrophy or ischaemic damage, were preserved or even enhanced by chronic heart rate reduction. The expansion of the capillary bed in the vascularly compromised heart induced by bradycardia may be stimulated by mechanical stretch of the endothelium and/or VEGF activated by chamber dilation and myocyte stretch. The increased number of capillaries and more homogeneous distribution of capillary perfusion would support the better pump function, even in the absence of higher coronary flow. The beneficial impact of chronic heart rate reduction on myocardial angiogenesis and function in cardiac hypertrophy and infarction may be major factor in the success of beta-blockers in treatment of human heart failure. PMID- 16308736 TI - Angiogenesis in the human heart: gene and cell therapy. AB - The concept of therapeutic angiogenesis -- stimulation of new vessels growth to restore blood supply to ischemic tissue has been studied in a number of clinical trials in patients with advanced coronary and peripheral arterial disease. This review discusses the main biological processes underlying new vessel growth and addresses applications of growth factor and cell therapy based on the stimulation of angiogenesis. While still very young and controversial, cell therapy has an enormous potential that is yet to be explored. Multiple questions remain unanswered including the choice of the best cell type, patient selection and the mechanism of action. Nevertheless, much should be expected in this area in the next decade with the likely emergence of new therapies for treatment of ischemic diseases. PMID- 16308757 TI - Germination of conidia and blastospores of Paecilomyces fumosoroseus on the cuticle of the silverleaf whitefly, Bemisia argentifolii. PMID- 16308763 TI - Infectious disease risk of cadaveric tissue donors who used non-injected illicit drugs. AB - This study assessed the correlation between a history of non-injection illicit drug use by cadaveric tissue donors and the presence of markers for Human Immunodeficiency Virus Type 1 (HIV-1), Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Human T Lymphocyte Virus Types I-II (HTLV-I-II), and Syphilis in their blood. It was found in 12,227 donors recovered in 2000-2002 that 9.68% of individuals with a history of non-injection drug use were seropositive for one or more infectious disease markers compared to 4.26% of donors without a history of drug abuse (p<0.001). A history of non-injection drug use had a positive predictive value of 9.68% and a negative predictive value of 95.74%. Analysis of seropositivity rates associated with individual drugs indicated that 9.6% of cocaine abusers were positive for one or more markers (p<0.001). Other drugs exhibited higher rates of seropositivity but the numbers were insufficient for statistical analysis. Other risk factors (transfusion of blood/blood products, tattoos and body piercing, incarceration) were not associated with higher incidence of infectious disease markers. PMID- 16308764 TI - Preventing disease transmission by deceased tissue donors by testing blood for viral nucleic acid. AB - Nucleic acid testing (NAT) has reduced the risk of transmitting infectious disease through blood transfusion. Currently NAT for HIV-1 and HCV are FDA licensed and performed by nearly all blood collection facilities, but HBV NAT is performed under an investigational study protocol. Residual risk estimates indicate that NAT could potentially reduce disease transmission through transplanted tissue. However, tissue donor samples obtained post-mortem have the potential to produce an invalid NAT result due to inhibition of amplification reactions by hemolysis and other factors. The studies reported here summarize the development of protocols to allow NAT of deceased donor samples with reduced rates of invalid results. Using these protocols, inventories from two tissue centers were tested with greater than 99% of samples producing a valid test result. PMID- 16308765 TI - Should deceased donors be tested for vCJD? AB - The 1997 Bovine Spongiform Encephalopathy enquiry and the 2001 Hepatitis C litigation judgement set the UK scene for evoking the precautionary principle and the legal precedent that liability for defective transfusion products should not be dependent on medical negligence, but on the mere fact of defectiveness. Animal models indicate that vCJD in humans, with infection via the oral route, is likely to be associated with infectivity within the lymphoreticular system (LRS). This is likely to appear prior to the involvement of the central nervous system and thus infectivity is likely to be present in the LRS before the onset of clinical disease. A number of relevant epidemiological studies using LRS tissue have shown a low, but measurable, existence of the carrier state in vCJD. Two possible cases of transmission of the abnormal prion of vCJD by blood transfusion suggested that tissues might also transmit and that testing of LRS tissue from deceased potential tissue donors should be considered as a first measure towards the prevention of vCJD transmission by tissues designated for use in transplantation. A variety of different tissues could be used as representative of the LRS, but each is associated with problems of feasibility and practicality. Assays for vCJD have not been validated in the context of donor screening rather than epidemiological studies nor on deceased donors. However, given the number of vCJD cases in the UK, significant attention should be paid to the logistical, ethical, social and other issues associated with undertaking vCJD testing of tissue donors, with a view to introducing testing of deceased tissue donors for vCJD disease or latency. PMID- 16308766 TI - Validating a low dose gamma irradiation process for sterilizing allografts using ISO 11137 method 2B. AB - This paper describes the validation of an allograft sterilization method specifically designed for the processing methods used at AlloSource in Centennial, CO. The methods used for this validation followed ISO Standard 11137, Method 2B. Three hundred allografts, collected from three defined production batches were dosed using a series of five incremental doses, beginning at 1 kGy and increasing by 1 kGy until 5 kGy was achieved. Following sterilization dosing, each allograft test article was analyzed using a sterility test to identify any viable microorganisms. The number of positive sterility samples was used to calculate the verification dose (1.27 kGy), which was then verified by an additional batch of 100 allografts. The results from this validation indicate that sterility (10(-6) SAL) on human allograft tissue using gamma 60Co radiation can be achieved when a dose of at least 9.2 kGy is employed. PMID- 16308767 TI - Processing of whole femoral head allografts: validation methodology for the reliable removal of nucleated cells, lipid and soluble proteins using a multi step washing procedure. AB - Following the major outbreak of BSE in the United Kingdom in the mid 1980's and the first reported cases of the human disease equivalent, vCJD, in 1996 a number of measures were taken to reduce the risk of secondary transmission of vCJD by blood or surgical instruments. Two cases of probable transmission of vCJD by blood components have now been reported. The Department of Health has recently undertaken a vCJD risk analysis for bone. Although significant uncertainties still exist in the assumptions on which the assessment is based, a few important recommendations were made. These include a recommendation that bone should be processed to remove blood and marrow and that pooling of donations is never advantageous. This study aimed to establish a method of processing whole femoral heads using disposable equipment, with the purpose of removing as much of the marrow components as possible. We are able to remove 98.2% (range 94.5-99.9%) of the nucleated cells and 98.7% (range 97.8-99.4%) of the soluble protein content from within an intact femoral head. This work confirms that the majority of blood and marrow components can be removed without the need for morselising the bone prior to washing. PMID- 16308768 TI - Effects of hydrogen peroxide cleaning procedures on bone graft osteoinductivity and mechanical properties. AB - Bone allografts are frequently used during orthopaedic trauma cases or other reconstructive procedures. Most allografts are processed and cleaned before use. Our goals were to determine if an improved cleaning procedure compromises the strength or osteoinductivity of a graft. We compared our improved cleaning procedure to our standard cleaning procedure on cortical bone allograft. The cleaning procedures are generally composed of a series of chemical steps with nonionic detergents, hydrogen peroxide, and alcohol under time and temperature control, subjected to ultrasonic agitation. We tested the compressive strength, impact strength, and shear strength following the standard and improved cleaning procedures. Osteoinductivity was tested in 4 groups, using the improved cleaning procedure with four different hydrogen peroxide cleaning times: 0, 1, 3, and 5 h. Osteoinductivity was evaluated in vivo, using a 28-day implant in the hamstring muscle of an athymic, nude mouse. Results demonstrated that osteoinductivity is maintained with cleaning in hydrogen peroxide for up to 1 h, and that compressive strength, impact strength, and shear strength were all unaffected by the improved cleaning procedure. The improved cleaning procedure therefore did not compromise the strength or osteoinductivity of cortical bone allografts in comparison to the standard procedure. PMID- 16308769 TI - Cryopreserved human B cells as an alternative source for single cell mRNA analysis. AB - Reverse transcription-polymerase chain reaction (RT-PCR) of individual B lymphocytes has been shown to be a powerful tool for the simultaneous analysis of different mRNA specificities in both malignant and non-malignant B cell subpopulations. However, especially for longitudinal studies, this may also require analyses of cryopreserved cells. Therefore, the current study assessed whether cryopreserved (liquid nitrogen, dimethyl sulfoxide [DMSO]-stored) viable B cells are an alternative source for single cell RT-PCR analysis. Fresh (non frozen) and post-thawed human peripheral blood B cells were analyzed by fluorescence-activated cell sorting (FACS). As a result, different B cell subpopulations could be reliably stained and separated from both fresh and post thawed cells by four-color flow cytometry, although slightly diminished fluorescence intensities of some subpopulation markers were observed when analyzing cryopreserved cells. Subsequently, viable individual CD19+CD27+ memory B cells were sorted into single wells and analyzed for the expression of mRNA transcripts of the 'house-keeping gene' glyceraldehyde phosphate dehydrogenase (GAPD), the constitutive B cell homing receptor CXCR4, and immunoglobulin heavy chain variable region (IgVH) genes by nested RT-PCR protocols. Comparing both B cell sources, RT-PCR analysis revealed comparable yields of cells expressing transcripts for the three mRNA specificities tested (GAPD, CXCR4, IgVH) indicating the integrity of the respective mRNAs in cryopreserved B cells. In conclusion, these data indicate that optimally cryopreserved B cells may be an alternative source for single-cell RT-PCR analysis, especially in longitudinal B cell studies. However, the settings for both FACS analysis and RT-PCR should be re-evaluated for each distinct subpopulation and target mRNA of interest when analyzing post-thawed cells. PMID- 16308776 TI - Evaluation score and interpretation index for the ecological quality of running waters in Central and Northern Hellas. AB - The present study aims at creating an evaluation system for the quality of running waters, based on the analysis of benthic macroinvertebrate records from Hellenic rivers (473 samples from 8 river basins). The proposed evaluation system (Hellenic Evaluation Score and its Interpretation Index) may be used for waters sampled with the cost effective semi-quantitative sampling method of "3 min kick sweep" and requires benthic macroinvertebrates to be identified to the taxonomic level of family. Though resulting from a modification of the Spanish score BMWP, it differs from it in the following characteristics: a) it includes the relative abundance of benthic macroinvertebrates and b) it takes into consideration the habitat diversity of the studied site, classifying it as "poor" or "rich", based on some parameters of the System B of the Water Framework Directive (2000/60/EU). Its interpretation is also based on a five-scaled classification system, consistent with the provisions of the same Directive. PMID- 16308777 TI - Animal hair as biological indicator for heavy metal pollution in urban and rural areas. AB - Animal hair is a good biomonitoring tool for heavy metals assessment and reflects the content of heavy metals in the forage and soil. Heavy metals Fe, Mn, Co and Ni as well as toxic metals Cd and Pb were determined in goat, sheep and camel hair, forage and soil collected from four different environmental urban and rural regions. These regions are Aswan city farms, Allaqi desert pasture, Kalabsha farms and Halaiub desert pasture at far south of Egypt. The results reveal that sheep hair contains the highest levels of Fe and Mn (879 and 55 microg/g, respectively), camel hair contains the highest Pb, whereas for goat Cd and Ni were the highest. Heavy metal concentrations in the studied hair reflect the presence of these metals in the surrounding forage and soil and vary from one area to another, and give knowledge of pollution in the area. Correlation statistics analysis and cluster analysis show a good and significant value between metals in hair and plants. PMID- 16308778 TI - The chemistry of soils, rocks and plant bioindicators in three ecosystems of the Holy Cross Mountains, Poland. AB - In June of 2000, biogeochemical study was carried out in three ecosystems of the Holy Cross Mountains (south-central Poland). This paper presents element concentrations and stable sulfur (and in one site lead) isotope ratios in rocks, detailed soil profiles, and plant bioindicators including epiphytic lichen Hypogymnia physodes (L.) Nyl., mosses (Pleurozium schreberi (Brid.) Mitt., Hypnum cupressiforme Hedw. S. Str., Hylocomium splendens (Hedw.) B.S.G.), Scots pine (Pinus sylvestris L.), common birch (Betula pendula Roth.), aspen (Populus tremula L.) and English oak (Quercus robur L.). Chemical analyses were performed with ICP-AES and AAS methods in the accredited laboratory (Central Chemical Laboratory of the Polish Geological Institute in Warsaw). The principal objective of this study was to compare the chemical composition of rocks, soils and selected plant bioindicators between investigation sites using the same methods of sample collection, preparation and analyses. The results of this study have shown that there is a high variability in concentrations of elements in plant bioindicators from the same sites that can not be explained only by soil properties or anthropogenic influence. This conclusion indicates that for biomonitoring purposes (especially with vascular plants) we cannot neglect individual features of the species examined. PMID- 16308779 TI - Derivation and use of sediment quality guidelines for ecological risk assessment of metals and radionuclides released to the environment from uranium mining and milling activities in Canada. AB - The Screening Level Concentration (SLC) approach was used to derive Lowest Effect Level (LEL) and Severe Effect Level (SEL) concentrations for nine metals (As, Cr, Cu, Pb, Mo, Ni, Se, U and V) and three radionuclides (226Ra, 210Pb, and 210Po) released to the aquatic environment during the mining and milling of uranium ore. This method was chosen because it allowed for the best use of the considerable historical and current data collected for diverse purposes in the uranium mining and milling regions of Canada (20,606 data points used in the analysis). Except for Cr, all the LELs derived in this study using the weighted method and published sediment quality guidelines (SQGs) were highly reliable (> 85%) in predicting sites unimpacted by uranium mining/milling defined as sites where reductions in the abundance and species richness of benthic invertebrate communities were < 20%. The derived SEL values and corresponding published SQGs (with the exception of Ni) were not reliable predictors (< or = 60%) of severe impacts on benthic invertebrate communities when severe impacts are defined as a reduction in abundance and species richness > or = 40%. Most of the severely impacted sites had sediment contaminant concentrations well below the SEL values. It is concluded that LELs derived using the weighted method can reliably be used in ecological risk assessments as concentrations below which adverse effects on benthic invertebrate communities are not expected. In contrast, it is recommended that SELs not be used in assessments of uranium mining/milling activities as concentrations above which adverse effects are anticipated. PMID- 16308780 TI - A comparison of three methods to collect submerged aquatic vegetation in a shallow lake. AB - Two boat-based and one in-water sampling method have been used to collect submerged aquatic vegetation (SAV) as part of a long-term monitoring program in Lake Okeechobee, Florida, USA. The boat-based methods consisted of collecting SAV with a ponar dredge, used only to collect Chara, and an oyster tongs-like rake apparatus, used to collect all SAV. The in-water method involved use of a 0.5 m2 PVC quadrat frame deployed by a diver. During summer 2002, SAV biomass samples were collected using all three methods at various sites in the lake to compare between-methods sampling precision. Sites used for these comparisons were selected based on plant type, plant density and sediment type. Statistical comparisons indicated that there were significant (p < or = 0.05) biomass differences in 8 of 15 possible pairwise comparisons between sampling method biomass means. In four of the eight comparisons, significantly higher biomass mean values were obtained using the quadrat frame. In three of the remaining four comparisons, significantly higher biomass mean values were obtained with the rake apparatus. For the fourth comparison, a significantly higher biomass mean value was obtained with the ponar dredge. Three of the four relationships between SAV biomass collected by the rake and the quadrat and the rake and the quadrat/ponar dredge were statistically significant, linear and explained between 67 and 78% of the biomass variability. There were no significant differences between regression coefficients or elevations for these relationships, therefore these regressions estimated the same population regression. The population regression coefficient was 0.95, suggesting that the quadrat and ponar over-sampled relative to the rake, but the amount of this over-sampling was very small. Since there was no consistent pattern in the sampling method which yielded the significantly different biomass values and there were no significant differences in sampling precision across a range of plant species, plant densities and two sediment types, the boat-based rake method appears to be a suitable replacement for the previously used ponar dredge and quadrat methods, when in-lake measurements are not practical. PMID- 16308781 TI - Monitoring of fogwater chemistry in the gulf coast urban industrial corridor: Baton Rouge (louisiana). AB - Seventeen fog events were sampled in Baton Rouge, Louisiana during 2002-2004 as part of characterizing wet deposition by fogwater in the heavily industrialized corridor along the Louisiana Gulf Coast in the United States. These samples were analyzed for chemical characteristics such as pH, conductivity, total organic and inorganic carbon, total metals and the principal ion concentrations. The dominant ionic species in all samples were NH4+, NO3-, Cl- and SO4(2-). The pH of the fogwater sampled had a mean value of 6.7 with two cases of acidic pH of 4.7. Rainwater and fogwater pH were similar in this region. The acidity of fogwater was a result of NO3- but partly offset by high NH4+. The measured gaseous SO2 accounted for a small percentage of the observed sulfate concentration, indicating additional gas-to-particle conversion of SO2 to sulfate in fogwater. The gaseous NOx accounted for most of the dissolved nitrate and nitrite concentration in fogwater. The high chloride concentration was attributable to the degradation of chlorinated organics in the atmosphere. The metal composition was traced directly to soil-derived aerosol precursors in the air. The major metals observed in fogwater were Na, K, Ca, Fe, Al, Mg and Zn. Of these Na, K, Ca and Mg were predominant with mean concentrations > 100 microM. Al, Fe and Zn were present in the samples, at mean concentrations < 100 microM. Small concentrations of Mn (7.8 microM), Cu (2 microM), Pb (0.07 microM) and As (0.32 microM) were also observed in the fogwaters, and these were shown to result from particulates (PM2.5) in the atmosphere. The contribution to both ions and metals from the marine sources in the Louisiana Gulf Coast was minimal. The concentrations of all principal ionic species and metals in fogwater were 1-2 orders of magnitude larger than in rainwater. Several linear alkane organic compounds were observed in the fogwater, representing the contributions from petroleum products at concentrations far exceeding their aqueous solubility. A pesticide (atrazine) was also observed in fogwater, representing the contribution from the agricultural activities nearby. PMID- 16308782 TI - Dynamic modeling of the ratio volatile fatty acids/bicarbonate alkalinity in a UASB reactor for potato processing wastewater treatment. AB - In this work, dynamic mathematical model for the prediction of the operational parameter volatile fatty acids/bicarbonate alkalinity (VFA/ALK) in a UASB reactor was developed. The dynamic modeling technique was applied successfully to a two year data record from an industrial wastewater treatment plant of a potato processing industry. The technique used included regression analysis by residuals. Seventeen parameters were examined including the following: wastewater's flow rate, reactor's temperature and pH, total and soluble influent COD, wastewater's temperature and pH, total and soluble effluent COD, volatile fatty acids, alkalinity, biogas production rate and each parameter with a time lag of up to 10 days. Finally, after all parameters and all time lag trials the best fitted model was developed. The model's adequacy was checked by chi2 test for a data record of the same UASB reactor but at a different time period and proved to be satisfactory. Additionally, the model's ability to predict and to control the plant's operation via VFA/ALK was examined. Through this model, in contrary to steady state models, various aspects of the process can be enlighten, such as the fact that the hydrolysis of starch requires at least a resident time of seven days. PMID- 16308783 TI - Extractable hydrocarbons, nickel and vanadium contents of Ogbodo-Isiokpo oil spill polluted soils in Niger Delta, Nigeria. AB - An oil spill polluted site at Ogbodo-Isiokpo in Ikwere Local Government Area of Rivers State in southern Nigeria, was identified for study following three successive reconnaissance surveys of oil fields in the Agbada west plain of Eastern Niger Delta. A sampling area of 200 m x 200 m was delimited at the oil spill impacted site using the grid technique and soils were collected at surface (0-15 cm) and subsurface (15-30 cm) depths from three replicate quadrats. A geographically similar, unaffected area, located 50 m adjacent to the polluted site, was chosen as a control (reference) site. Total extractable hydrocarbon contents of the polluted soils ranged from 3.02-4.54 and 1.60-4.20 mg/kg (no overlap in standard errors) at surface and subsurface depths respectively. The concentrations of two "diagnostic" trace heavy metals, nickel (Ni) and vanadium (V), which are normal constituents of crude oil, were also determined in the soils by atomic absorption spectrophotometric method after pre-extraction of cations with dithionite-citrate carbonate. Ni varied from 0.15 to 1.65 mg/kg in the polluted plots and from 0.18 to 0.82 mg/kg in the unpolluted plots; vanadium varied from 0.19 to 0.70 mg/kg in the polluted plots and from 0.14 to 0.38 mg/kg in the unpolluted plots. Ni and V were more enhanced (p < 0.05) in the oil polluted soils, especially at subsurface depth. Whilst the oil spillage could be said to be indirectly responsible for the enhanced concentrations of nickel and vanadium via the injection and availability of the petroleum hydrocarbons that might have increased the activities of biodegradation on site, the physico chemical properties of the soils and inherent mobility of metals, as well as the intense rainfall and flooding that characterized the period of study, may have also contributed, at least in part, to these enhanced concentrations. Such levels of Ni and V may result to enhanced absorption by plants, which may bring about possible bioaccumulation in such plants and the animals that depend on them for survival and all of these may lead to toxic reactions along the food chain. PMID- 16308784 TI - Exploring the relationship between hydrologic parameters and nutrient loads using digital elevation model and GIS - a case study from Sugarcreek headwaters, Ohio, U.S.A. AB - Ohio is typical among the Midwestern and Eastern United States with high levels of water pollutants, the main sources being from agriculture. In this study, we used a digital elevation model in conjunction with hydrological indices to determine the role of landscape complexity affecting the spatial and temporal variation in pollutant levels, in one of the most impaired headwater streams in Ohio. More than eighty five percent of the study area is dominated by agriculture. Spatial distribution of slope (S), altitude and wetness index along with other watershed parameters such as flow direction, flow accumulation, stream networks, flow stream orders and erosion index were used within a Geographic Information Systems framework to quantify variation in nitrate and phosphate loads to headwater streams. Stream monitoring data for nutrient loads were used to correlate the observed spatial and temporal patterns with hydrological parameters using multiple linear regressions. Results from the wetness index calculated from a digital elevation model suggested a range of 0.10-16.39, with more than 35% having values less than 4.0. A Revised Universal Soil Loss Equation (RUSLE) predicted soil loss in the range of 0.01-4.0 t/ha/yr. Nitrate nitrogen levels in the study area paralleled precipitation patterns over time, with higher nitrate levels corresponding to high precipitation. Atmospheric deposition through precipitation could explain approximately 35% of total nitrate levels observed in streams. Among the different topographic variables and hydrological indices, results from the step-wise multiple regression suggested the following best predictors, (1) elevation range and upstream flow length for nitrate, (2) flow direction and upstream flow length for ammonia-nitrogen and slope, and (3) elevation range for phosphate levels. Differences in the landscape models observed for nitrate, phosphate and ammonia-nitrogen in the surface waters were attributed partly to differences in the chemical activity and source strengths of the different forms of these nutrients through agricultural management practices. The results identify geomorphologic and landscape characteristics that influence pollutant levels in the study area. PMID- 16308785 TI - Use of Central stonerollers (Cyprinidae: Campostoma anomalum) from Tennessee as a bioindicator of metal contamination. AB - We compared the levels of arsenic, cadmium, chromium, lead, manganese, mercury and selenium in a small species of fish (Central stonerollers, Campostoma anomalum) collected from East Fork Poplar Creek (EFPC) and a reference site in eastern Tennessee. Stonerollers are minnows in the Cyprinidae family that serve as prey for many carnivores in aquatic systems. Fish were collected from East Fork Poplar Creek within the U.S. Department of Energy's Y-12 National Security Complex, part of the Oak Ridge Reservation, and from a reference stretch of the Little River in East Tennessee. Whole fish were homogenized for analysis. Concentrations of all metals (except arsenic) were significantly higher in stonerollers from EFPC compared to the reference site. Mercury levels in minnows from EFPC averaged 0.4 ppm (microg/g), four times higher than the average for fish in the U.S. in general. This was higher than levels in fish from the nearby Clinch River and higher than fillets of white bass (Morone chrysops) from the same creek. Most metal levels were inversely related to size and weight of the stonerollers, perhaps due to growth dilution. PMID- 16308786 TI - Contamination of oxygen-consuming organics in the Yellow River of China. AB - Contamination of oxygen-consuming organics (OCOs) was one of the most serious problems in the Yellow River of China. This study was conducted to analyze monitoring of the data on OCOs contamination for the river in 1980 and during 1992-1999 as well as examining the effect of suspended solids (SS) on chemical oxygen demand (CODMn) and biochemical oxygen demand (BOD5) of river water. Several significant results have arisen from the study. First, CODMn and BOD5 of the river water showed an increasing trend from the upper to the lower reaches of the mainstream. BOD5 values of river water in 1992 were significantly higher than those in 1980 and showed an increasing trend during 1992-1999. Second, OCOs in river water of the mainstream was attributed mainly to point sources; the ratio of point to non-point sources of BOD5 was about 2.81. The load from point sources showed an increasing trend during 1992-1998. In contrast, the load from non-point sources manifested a decreasing trend during this period; this was caused by the decreasing trend of SS content in river water. The total load of BOD5 from point and non-point sources displayed an increasing trend during 1992-1998. Third, as the humic substances in SS can hardly be biologically oxidized in natural conditions but can be oxidized by chemical oxidants such as potassium permanganate, CODMn was not suitable for being regarded as a parameter reflecting the pollution degree of OCOs in river water with a high SS content. PMID- 16308787 TI - Patchy distribution fields: an interleaved survey design and reconstruction adequacy. AB - A mathematical model was used to compare the effects of a regular (one-pass) or interleaved (two-pass) acoustic survey on the adequacy of reconstructing patchy distribution fields. The model simulates fish or plankton patches of different shapes and spatial orientations, and a set of parallel or zigzag transects forming a regular or interleaved acoustic survey. The efficiency of a survey is determined by the adequacy of a reconstructed field to that originally generated, which is evaluated by calculating their correlations. Regarding the immovable fields, the efficiency of a regular or interleaved acoustic survey was tested with the following two alternative assumptions: (1) the entire survey was completed; (2) the survey was interrupted for some reason at the moment when one transect remained non-accomplished. In the former case, the efficiencies of both acoustic surveys were nearly the same; in the latter case, the efficiency of an interleaved survey was superior to that of a regular one. With respect to movable fields, the efficiency of the completed interleaved surveys was even higher than that of the regular ones. Thus, the results obtained allow us to conclude that an interleaved survey is expedient in cases where there is no preference regarding the position of a vessel for further work. PMID- 16308788 TI - Profile of PAH in the exhaust of gasoline driven vehicles in Delhi. AB - A preliminary study to determine the profile of PAHs in the exhaust of gasoline vehicles in Delhi was conducted. Three different types of vehicles (cars, autorickshaws and scooters) were selected with different age groups for sampling purpose. The concentration of Total PAHs (Sigma12PAHs) was found to be 27.27 +/- 2.27, 28.61 +/- 3.70 and 29.81 +/- 3.57 mg/g in the exhaust of cars, auto- rickshaws (three wheelers) and two wheelers, respectively. The levels of PAHs were found to be high in scooter exhaust as compared to that of cars and autorickshaws. The total PAHs concentration in the present study was found to be higher as compared to other studies. Such a high concentration could be attributed to different parameters like the age of the vehicles, driving conditions, the fuel quality and the emission standards. PMID- 16308789 TI - Monitoring of criteria air pollutants in Bursa, Turkey. AB - The concentrations of criteria air pollutants such as CO, NOx (NO + NO2), SO2 and PM were measured in the period of May 2001 and April 2003 in the city of Bursa, Turkey. The average concentrations for this period were 1115 +/- 1600 microg/m3, 29 +/- 50 microg/m3, 51 +/- 24 microg/m3, 79 +/- 65 microg/m3, 40 +/- 35 microg/m3, 98 +/- 220 microg/m3, for CO, NO, NO2, NOx, SO2 and PM, respectively. Temporal changes in concentrations were analyzed using meteorological factors. Correlations among pollutant concentrations and meteorological parameters showed weak relations nearly in all data. Lower concentrations were observed in the summer months while higher concentrations were measured in the winter months. The increase in winter concentrations was probably due to residential heating. Pollutants were associated with each other in order to have information about their origin. NOx/SO2 ratio was also examined to bring out the source origin contributing on air pollution (i.e., traffic or stationary). PMID- 16308790 TI - Impact of post-methanation distillery effluent irrigation on groundwater quality. AB - Molasses-based distilleries generate large quantities of effluent, which is used for irrigation in many countries including India. The effluent is rich in organic and inorganic ions, which may leach down and pollute the groundwater. An on-farm experiment was conducted to assess the impact of long-term irrigation with post methanation distillery effluent (PMDE) on nitrate, sulphate, chloride, sodium, potassium, and magnesium contents in the groundwater of two sites in northwest India. Electrical conductivity (EC), pH, total dissolved solids (TDS), sodium adsorption ratio (SAR) and colour were also determined to assess the chemical load in the groundwater. Nitrate content in the groundwater samples ranged from 16.95 mg L(-1) in the unamended fields to 59.81 mg L(-1) in the PMDE-amended fields during the 2-year study (2001-2002). Concentrations of TDS in water samples from tubewell of the amended field was higher by 40.4% over the tubewell water of the unamended field. Colour of the water samples of the amended fields was also darker than that of the unamended fields. The study indicated that the organic and inorganic ions added through the effluent could pose a serious threat to the groundwater quality if applied without proper monitoring. PMID- 16308791 TI - Evaluation of lead availability in amended soils monitored over a long-term time period. AB - Two different soil amendment processes were evaluated for reducing lead availability from a contaminated soil at a demonstration study site, to reduce potential public health and environmental concerns. A limited variety of in vitro laboratory "availability" tests (relative bioaccessible and environmental mobility) were performed to determine if the available lead in the contaminated soil would be less available after in situ soil amendment (chemical treatment). The relative bioaccessibility results were evaluated in both a short-term period (within 24 h after treatment) and over a long-term time period (quarterly basis for 5 years). Reduction in relative bioaccessibility was noted for one of the treatments immediately after treatment; however, both treatments indicated a significant upward trend in bioaccessibility values over a 5-year time period after treatment. The comparison between the treated units and the control units indicated that the long-term effectiveness of the treatment processes could not be demonstrated. PMID- 16308792 TI - Heavy metal transport and behavior in the lower Columbia River, USA. AB - The primary objective of this study was to evaluate temporal changes in heavy metal content of lower Columbia River sediment following terminated or reduced soluble heavy metal loading from the world's largest lead-zinc refinery and mining districts in the USA and Canada. Sediment cores were collected from two fine sediment depositional sites ( approximately 600 km downstream) in August 1999 and were analyzed for total metal content, texture, and age/dating parameters. Zinc, cadmium and lead contents in 1999 declined by only a factor of two over their depth profile maxima (dated as between 1970 and 1980). In sharp contrast, more than a 10-fold decrease in dissolved metal loading occurred during this same period. Zinc in filtered Columbia River water at downstream locations also declined by > 10-fold, consistent with the reduced upper river solute-metal loading. Once soluble metal releases are reduced or terminated, the solute half time in Columbia River water is months versus approximately 20 yr for adsorbed metals on surficial (or resuspended) bed sediments. The much slower rate of decline for sediment, as compared to the solute phase, is attributed to resuspension, transport and redeposition of irreversibly bound metals from upstream sedimentary deposits. This implies downstream exposure of benthic or particle-ingesting biota can continue for years following source remediation and/or termination of soluble metal releases. Accordingly, contaminant contents of both particulate and solute phases of river water, as well as sediment core sections, are suggested for assessing long-term biotic exposure/response to mitigation activities in the Columbia River and similar fluvial ecosystems. PMID- 16308793 TI - Automated classification of an environmental sensitivity index. AB - Environmental Sensitivity Indices (ESI) composed of many field-data are essential for monitoring and control systems. At the beginning of the last decade an ESI of the German Wadden Sea was developed for use by the relevant authorities. This ESI was derived by experts semi-manually analysing the extensive field data-set. An algorithm is presented here which emulates human expert-decisions on the classification of sensitivity classes. This will permit the necessary regular updates of ESI-determination when new field data become available using automated classifications procedures. After tuning the algorithm parameters it generates decisions identical to those of human experts in about 97% of all locations tested. In addition, the algorithm presented also enables erroneous or extremely seldom field data to be identified. PMID- 16308794 TI - Evaluation of water quality in the Chillan River (Central Chile) using physicochemical parameters and a modified water quality index. AB - The Chillan River in Central Chile plays a fundamental role in local society, as a source of irrigation and drinking water, and as a sink for urban wastewater. In order to characterize the spatial and temporal variability of surface water quality in the watershed, a Water Quality Index (WQI) was calculated from nine physicochemical parameters, periodically measured at 18 sampling sites (January November 2000). The results indicated a good water quality in the upper and middle parts of the watershed. Downstream of the City of Chillan, water quality conditions were critical during the dry season, mainly due to the effects of the urban wastewater discharge. On the basis of the results from a Principal Component Analysis (PCA), modifications were introduced into the original WQI to reduce the costs associated with its implementation. WQIDIR2 and WQIDIR, which are both based on a laboratory analysis (Chemical Oxygen Demand) and three (pH, temperature and conductivity), respectively, four field measurements (pH, temperature, conductivity and Dissolved Oxygen), adequately reproduce the most important spatial and temporal variations observed with the original index. They are proposed as useful tools for monitoring global water quality trends in this and other, similar agricultural watersheds in the Chilean Central Valley. Possibilities and limitations for the application of the used methodology to watersheds in other parts of the world are discussed. PMID- 16308795 TI - Laboratory-scale chlorination to estimate the levels of halogenated DBPs in full scale distribution systems. AB - Occurrence of halogenated disinfection by-products (DBPs) (trihalomethanes -THMs- and haloacetic acids -HAAs-) in the waters of two utilities in Quebec City (Canada) was investigated using two approaches: experimental chlorination studies and full-scale sampling within distribution systems. Experimental studies were designed to reproduce treatment plant and distribution system conditions (chlorine dose, water temperature, pH and water residence time). Differences in halogenated DBPs in the two distribution systems under study were significant and comparable to those observed in experimental laboratory studies. For the waters of both utilities, chlorination studies better reproduced the occurrence of halogenated DBPs in points of the distribution system located near the treatment plant (low residence time of water) than in other points. Multivariate regression models for THMs, HAAs and their species were developed using the data from experimental studies in order to predict halogenated DBP levels measured in the distribution system. Models were all statistically significant, but showed low ability to predict full-scale halogenated DBPs, particularly in points located at distribution system extremities. Specifically, experimental chlorination-based models are not able to simulate the decrease of HAA levels. Results of this research suggest that the use of experimental data to predict halogenated DBP levels in full-scale distribution systems - for operational, regulatory and epidemiological purposes - must be done with caution. PMID- 16308796 TI - Monitoring of organochlorine pesticide residues in summer and winter vegetables from Agra, India - a case study. AB - Analysis of summer and winter vegetable samples during 2002-2003 for pesticidal contamination was carried out on Gas Chromatograph-Electron Capture Detector with capillary columns. The contamination levels of winter vegetables (average concentration of 4.57, 6.80 and 5.47 ppb respectively for Lindane, Endosulphan and DDT) were found to be slightly higher than the summer vegetables (average concentration of 4.47, 3.14 and 2.82 ppb respectively for Lindane, Endosulphan and DDT). The concentration of these organochlorine pesticides in summer and winter vegetables were well below the established tolerances but continuous consumption of such vegetables even with moderate contamination level can accumulate in the receptor's body and may lead to chronic effects that could be fatal. PMID- 16308798 TI - Is endoscopic totally extraperitoneal hernioplasty justified for the repair of groin hernia in female patients? AB - BACKGROUND: Groin hernia is an uncommon surgical pathology in females. The efficacy of the endoscopic approach for the repair of female groin hernia has yet to be examined. The current study was undertaken to compare the clinical outcomes of female patients who underwent open and endoscopic totally extraperitoneal inguinal or femoral hernioplasty (TEP). METHODS: From July 1998 to June 2004, 108 female patients who underwent elective repair of groin hernia were recruited. The patients were divided into TEP (n = 30) and open groups (n = 78) based on the type of operation. Clinical data and outcome parameters were compared between the two groups. RESULTS: The mean ages and hernia types were comparable between the two groups. All TEPs were successfully performed. The mean operative times were 52 min for unilateral TEP and 51 min for open repair. The difference was not statistically significant. Comparisons of the length of hospital stay, postoperative morbidity, pain score, and time taken to resume normal activities showed no significant differences between the two groups. A single patient in the TEP group experienced recurrence of hernia. CONCLUSIONS: The findings show equivalent postoperative outcomes after TEP and open repair of groin hernia in female patients. Because the wound scar after open repair is well concealed beneath the pubic hair and no superior clinical benefits are observed after TEP, open repair appears to be the technique of choice for the management of primary groin hernia in females. The TEP approach should be reserved for female patients with recurrent or multiple groin hernia. PMID- 16308799 TI - [Is mental health services research in need of randomised controlled trials?]. PMID- 16308800 TI - [For and against: broader indication of electroconvulsive therapy]. PMID- 16308801 TI - [Suicidal tendencies of physicians -- an overview]. AB - QUESTION: This study provides an overview of the research on suicide rates and suicidal tendencies of physicians. METHODS: Original articles on this topic since 1980 were analyzed. RESULTS: Empirical studies indicate an increased suicide rate in physicians compared to the general population. Possible causes are the increased prevalence rate of depressive disorders as well as the presumably increased rate of substance abuse in physicians. In addition to this, job-related stressors as reasons for the high suicide risk in physicians are discussed. CONCLUSION: In terms of preventive measures it seems reasonable to systematically inform medical students about stressors they will be exposed to in their later practice and to discuss coping strategies and prevention. Thus, a suitable emotional preparation for future work stress can be assured. For working physicians seminars and professional training on education and prevention as well as supervision should continuously be offered. PMID- 16308802 TI - [Job satisfaction and burnout in professionals in austrian mental health services]. AB - OBJECTIVE: The aim of this study was to explore job satisfaction, professional role and burnout among community mental health staff in Austria. METHODS: The Minnesota Job Satisfaction Questionnaire, the GHQ-12, the Maslach Burnout Inventory and a questionnaire exploring staff's professional role and team identity were administered to 195 community-based mental health professionals in two Austrian regions. Staff's job perception was assessed in open questions. Predictors of burnout scores were identified in multivariate analyses. RESULTS: While the mental health professionals had elevated scores on the GHQ-12, their levels on the three burnout subscales were low. Social workers and psychologists showed the lowest job satisfaction, social workers also had low role identity scores. Being in the current job for a short time and disposing of psychosocial skills were predictors of high job satisfaction. High burnout scores were predicted by the lack of basic psychosocial competence and a lack of general knowledge in mental health care. The effect of caseload on psychological well being was positive as well as negative. CONCLUSIONS: Even if the mental health professionals in our study show high levels of general stress, they seem to have less emotional problems resulting from extensively dealing with troubled individuals. Improving basic psychosocial competence and general expertise in mental health care might have a protective effect against developing a burnout syndrome. PMID- 16308803 TI - [COMSKILLS: assessing community living skills in offender groups]. AB - OBJECTIVE: An international network study involving parallel, complementary interventions in three EU countries and an associate country (Germany, United Kingdom, The Netherlands and Norway) is reported. The aim is to develop a unified approach to the assessment of social risk and related behaviours in offender groups. METHOD: The Behavioural Status Index (BEST-Index) and a set of cross validating instruments (PCL-R, HCR-20; SCL-90-R; BDHI-D) were applied to a sample of n = 231 mentally ill offenders. RESULTS: Selected results using a sub-sample of n = 89 German offenders are reported and discussed. CONCLUSIONS: The BEST Index shows convergent validity with respect to a social risk criterion and it helps to determine an objective data base for the improvement of carer assessments, related care planning, and delivery. PMID- 16308804 TI - [Is there currently a boom of stigma research in psychiatry?]. AB - OBJECTIVE: Recently, there has been a growing awareness among psychiatrists of the stigma of mental illness. In numerous countries programs aimed at reducing stigmatization and discrimination because of mental illness have been launched. The question arises as to what extent the increasing interest in stigma has stimulated research in this area. METHOD: A search for papers on stigma of mental illness that have been published in scientific journals since 1990 has been carried out, using different search strategies. RESULTS: During the 1990 s, there is a marked increase of articles dealing with the stigma of mental illness. There is no indication of an end of this trend. DISCUSSION: Despite more research we are still far from understanding the stigma process in detail. This, however, is the prerequisite for developing successful anti-stigma interventions. PMID- 16308805 TI - [Continuation ECT]. AB - A renaissance of electroconvulsive therapy in psychiatry can be observed in Germany. Here, ECT was named as a first line therapy for treating psychotic depression, depressive stupor, schizoaffective psychoses with severe depressive symptoms. Suprisingly, ECT is most commonly not used as a continuation therapy after achieving acute remission. With rare exceptions, antidepressive medication is chosen for this purpose. The use of continuation ECT (cECT) and subsequent maintenance ECT (mECT) is not (or just marginally) mentioned in practice guidelines. In our case report we report a successful cECT and mECT of a geriatric patient with severe comorbidity. This case is exemplified with respect to recent guidelines and study results. Particularly suggestions for interval duration, total duration of treatment, effects on cognition and memory, comorbidity and comedication, and reflections on quality of life and costs were discussed. We would therefore recommend a broader use of this proven treatment tool for keeping major depressions in remission. PMID- 16308806 TI - [Concepts for a demand-oriented structuring of palliative care in the German Health Care System: the example of Lower Saxony]. AB - Palliative care can be considered a holistic care approach for patients faced with incurable and progressive diseases in their last period of life. In this context, structural, cultural and content-related deficits are increasingly taken into account in the German health care system, however, there is a lack of scientific data. Therefore, it was the objective of an expert team at Hannover Medical School to analyse the current situation of palliative care in Lower Saxony from the perspective of health system research and to give recommendations for its further development, based on empirical studies in Lower Saxony and on a systematic national and international literature review. The study revealed that the main shortcomings were an unclear structuring and an inadequate definition with respect to other areas of care, a fundamental lack of integration in regard of routine processes of care and a lack of thorough tools for needs assessments. Therefore, a concept for the integration of palliative care in the care processes of cancer patients was developed and the specific need for hospice beds in regional districts was calculated by using parameters representing provision related and epidemiological/demographic conditions. In comparison to current structures, an assessment of over- and undersupply as well as specific recommendations how to optimize palliative care were provided for politicians, payers and providers in Lower Saxony. PMID- 16308807 TI - [A patient flow analysis within the Munich addiction treatment system--a pilot study]. AB - BACKGROUND: The highest efficiency in addiction treatment is attributed to a multistage programme consisting of several intervention types (e. g., counselling, withdrawal, therapy, community adjustment) in a specific order. AIM: The pilot study described focuses on the interactions between treatment modalities (i. e. on the process quality) within the addiction treatment system in Munich and the broader environs. METHOD: In eleven institutions out of the several intervention types 452 persons were anonymously queried. The interview was conducted by means of a questionnaire, which gathered information about the sequence of interventions. RESULTS: The results suggest that the treatment sequences (i. e. the patient flow) investigated are consistent with the intended cooperation within the multistage treatment programme. Furthermore, the results also indicate that more than a quarter of the participants came sought treatment/counselling of their own initiative. Particularly interesting is the observation that the participating institutions rarely checked where "their" patients pursue aftercare/treatment follow-up upon completion of the prescribed programme. Regarding the actual referrals during the four week investigation period, relatively low referrals among the participants in the drop-in center were observed. CRITICISM: It remains doubtful whether the selected institutions are really representative of the different intervention types, because the participating institutions within an intervention type operate from diverse conceptual backgrounds. Furthermore, the sample size obtained was both small and contained a diverse number of participants per institution type. Therefore, whether or not the results can be generalized is questionable. CONCLUSION: Based on these results the BAS supports the deployment and legal establishment of case managers in the coordination of a patient's global treatment process. It is believed that this step would result in a more purposeful and economical treatment within the multistage addiction treatment programme. PMID- 16308808 TI - [Doing voluntary extra work? Organizational citizenship behavior in the hospital- a comparison between physicians and nurses]. AB - AIM OF THE STUDY: The study compares physicians and the nursing staff of a hospital in terms of their extra-role behavior. Matters of interest include the extent of Organizational Citizenship Behavior (OCB) shown on the one hand and on the other hand which conditions stimulate the OCB of both physicians and nurses, respectively. METHOD: The comparison was conducted by applying a questionnaire on n = 70 physicians and n = 112 nurses in a nursing department of a municipal hospital. RESULTS: The results can be summarized as follows: (1) The extra-role behavior in terms of sportsmanship, individual initiative, and conscientiousness show equally high values with respect to physicians as well as nurses. In contrast, nurses rate their own helping behavior towards colleagues higher than the physicians do. Therefore, the extent of OCB does not seem to be job-specific in the narrower sense. (2) Differences between physicians and nurses exist indeed with respect to the conditions for the occurrence of OCB: Although the extent of OCB shown by physicians and nurses is independent from age, department tenure, and organizational tenure, job experience does play a role for the degree of conscientiousness (physicians) and individual initiative (nurses). Furthermore, gender affects the sub dimension sportsmanship (nurses). (3) While job characteristics (job control and stress) play a certain role for the degree of nurses' OCB, the physicians' extra-role behavior is independent from job control and strain. Vice versa, the analyzed person-related characteristics job insecurity and strain play a role for the extra-role behavior of physicians, while the behavior of nurses remains unaffected hereof. In other words: Nurses show the same OCB at high and low levels of strain and job insecurity, while physicians lower their OCB when strain and job insecurity rise. (4) For both physicians and nurses, job satisfaction is the most important predictor for extra role behavior. CONCLUSION: When trying to enhance the extent of OCB within a hospital, it is -- according to our results -- primarily essential to increase the job satisfaction of physicians as well as nurses. Within the nursing department, it is additionally recommended to enhance the employees' scope of action, if possible. However, for the enhancement of OCB it must be kept in mind - according to our results -- that with rising OCB the stress (e. g. time pressure and interruptions) rise at the same time. The latter might result in higher strain for employees. In the group of physicians, on the other hand, a person-related approach seems promising: it is essential to reduce the physicians' subjectively felt strains as well as the job insecurity. PMID- 16308809 TI - [Language promotion in kindergartens in Biberach county--an analysis of the state foundation subsidized language intervention project]. AB - The Programme Sprachforderung im Kindergarten was carried out in 83 nursery schools in the Landkreis Biberach/Germany from fall 2003 till summer 2004. The Programme was offered by the Kreisgesundheitsamt des Landkreises Biberach in cooperation with the Sozial- und Jugendamt and the Hor-Sprachzentrum Ravensburg and contained a language-intervention programme and a training for the nursery school teachers. Language screenings where conducted at the beginning and the end of the programme including 1479 Children of the age of 4 - 6 years. 23 % of the native German and 62 % of the migrant children showed a need for intervention. These children where fostered with the Ravensburger Modell for 6 months. The increase in performance was highly significant. Nevertheless the fostered children could not achieve the performance level of the native German children without language retardation with the exception of two tasks (word families and understanding of questions). The most significant increase in performance showed the poorest performers of the intervention-group. According to the increase in performance the programme was successful, but also obviously too short for the poorer performers to catch up. Therefore we advise further language-intervention programmes in nursery-schools to be conducted as early as possible. Furthermore analytical observation and supervision must be standard to maintain a high level of effectiveness. PMID- 16308810 TI - [Speech comprehension and speech usage in hearing impaired children--an empirical follow-up study]. AB - In a mean follow-up duration of 19 months the oral language development of 18 sensorineural hearing-impaired children without multiple handicaps was serially measured at regular intervals (three time points t1 - t3) using the receptive and expressive language scales of the "Munchener Funktionellen Entwicklungsdiagnostik fur das 2. und 3. Lebensjahr". Results are presented as developmental quotients (DQ). In case of language retardation the rate of language development is less than one. At t1, the average language development was considerably delayed (DQ receptive: 0.37; DQ expressive: 0.51) and slowly increased over time (receptive: 0.37/0.48/0.55; expressive: 0.51/0.51/0.55) at approximately half the rate of normal hearing children. With the hearing-impairment increasing, the DQs became smaller. Children who were identified as hearing-impaired before the age of 18 months exhibited on average generally greater DQs. A significant impact of speech/language therapy on language development at t3 could not be demonstrated. Finally, the results and health care requirements were discussed. PMID- 16308811 TI - [The vaccination coverage among children aged 24 - 30 months in pediatric offices November 1999 - May 2001. The influence of socio-demographic characteristics]. AB - BACKGROUND: The goal of this study was to investigate the influence of socio demographic factors on vaccination coverage of 24-30 month old children in paediatric practices. METHODS: Vaccination coverage (VC) was documented for 15,682 children in 196 paediatric practices mainly in the region of Nordrhein. Data on socio-demographic factors were collected for 8,457 children and their influence on the vaccination status (VS) was investigated by means of logistic regression analysis. RESULTS: Complete age-appropriate VC for all vaccines recommended by the Standing Committee for Vaccination (STIKO) was 49.9 %. Complete VC for Diphtheria/Tetanus (DT) was 74.6 % and for first dose measles/mumps/rubella (MMR), 82.1 %. The following factors were associated with complete VS for DT and MMR: completion of last recommended well-child visit, parents living together, low number of siblings, short time to reach practice, parents stating they felt adequately informed about vaccinations. Complete vaccination for DT was inversely associated with being health insured through social security and for MMR with the mother having graduated from high school. CONCLUSIONS: A high proportion of 24-30 month old children did not have an age appropriate VS. Targeted parental education and timely vaccination strategies that take into account socio-demographic risk factors for low vaccination uptake are required. PMID- 16308812 TI - [Smoking as potential confounder in german epidemiological studies: standardization of assessment, quantification, and analysis]. AB - Tobacco smoking is one of the most frequently examined risk factors in epidemiological studies due to the high frequency of smoking in the population and its high risk potential. This paper discusses the issue of standardization of the assessment of tobacco exposure in German epidemiological studies based on a suggestion of the working group "Epidemiological methods" of the German Society of Epidemiology (DGEpi), the German Society of Social Medicine and Prevention (DGSMP), and German Society of Medical Informatics, Biometry and Epidemiology (GMDS) together with the working group "Epidemiology in the workplace" of the DGEpi and the German Society of Occupational and Environmental Medicine (DGAUM). The main element is a short questionnaire for German adults for the assessment of tobacco smoking in epidemiological studies where smoking is a potential confounder. In addition, information on the consideration of intermittent periods of non-smoking, a very short questionnaire for medical examinations in the workplace or non-responder interviews as well as the quantification and statistical modelling of smoking are presented. A brief discussion of scientific problems and questionnaires related to the exposure to environmental tobacco smoke is given. PMID- 16308813 TI - [Screening for colorectal cancer: what is the most effective strategy?]. AB - Colorectal cancer is one of the most common cancers in Western countries. The long natural history of colorectal cancer provides opportunities for detection of early stage cancer and for prevention of cancer by removal of adenomas. An effective screening technique should significantly reduce morbidity and mortality in the population. Several tests are available to screen healthy subjects at average risk for colorectal cancer: total colonoscopy, combined sigmoidoscopy and faecal occult blood test (FOBT), FOBT alone. At the present time clear evidence for the most effective screening strategy is not available. Colonoscopy screening in Germany needs an adequate tool to evaluate the programme. Follow up is essential and cancer registries would be an important partner. PMID- 16308814 TI - [10-year PCB Guideline--attempt to balance hygienic and medical/environmental toxicological views]. AB - PURPOSE: In 1995 a guideline was established in Germany, which requires the remediation of buildings with elevated indoor air concentrations of polychlorinated biphenyls (PCB). The action limit is 3000 ng/m(3). The tolerable PCB indoor air concentration is 300 ng/m(3). The aim of remediation actions is to reduce PCB in indoor air to levels < 300 ng/m(3) by removing the sources of PCB in buildings (mainly sealants, ceiling plates, wall colours). Based on this guideline a great number of school buildings, university buildings and administration buildings constructed in the 1960-ties and 1970-ties were renovated. In this paper, the toxicological basis of the aforementioned guide values are discussed. Furthermore, recent papers and proposals to establish lower guideline values for PCB in indoor air are discussed. CONCLUSIONS: The existing PCB guideline should be revised. When revising the guide values for PCB in indoor air it should be kept in mind that human PCB exposure is mainly via food, that PCB intake via food has been declining since more than 20 years, and that there is only a minor increase of PCB blood levels following inhalation of PCB in indoor air. In public discussions it should be clearly stated that remediation of PCB contaminated buildings is a preventive action. PCB concentrations in indoor air, which are associated with significant health risks, are presumably much higher than the current action value. PMID- 16308815 TI - [Periorbital contact eczema]. AB - Periorbital contact eczema is most commonly the result of an allergic contact dermatitis whereas other eczematous skin diseases like atopic eczema or seborrheic eczema occur less frequently. Also, other diseases like autoimmune disorders or rosacea need to be considered. Allergic contact dermatitis is a T cell-mediated immunological response towards ubiquitous contact allergens. Activated T-cells migrate through the vessels into the skin and produce several inflammatory mediators. Epicutaneous patch testing is an important tool for the diagnosis of contact allergy whereby the allergens are analysed in terms of their ability to induce eczematous skin reaction. Until now the short-term use of corticosteroids are is employed for the treatment of allergic contact eczema. Modern substances with an optimal therapeutic index should rather be used. PMID- 16308816 TI - [Imaging of the anterior eye chamber with optical coherence tomography]. AB - BACKGROUND: Optical coherence tomography (OCT) represents a high-resolution diagnostic method which can be used to precisely image the anterior eye segment. Further developments in scanning technology and data evaluation allowed to visualize the entire anterior eye segment in one image, and the purpose of this study was to clinically evaluate this new system. PATIENTS AND METHODS: The studied OCT system was adapted to a slit-lamp and allowed us to image the anterior segment with an axial resolution of 10 microm at a scanning rate of 200 Hz with a depth of 7 mm and a maximal width of 15 mm. In selected cases the clinical value of anterior segment OCT was assessed. The reproducibility and level of agreement of anterior chamber depth measurements were assessed during the preoperative examination in 49 cataract surgery patients and compared to ultrasound (US) biometry (10 MHz). RESULTS: The studied anterior segment OCT allowed the cross-sectional imaging of the entire anterior eye segment. Changes of the anterior chamber before and after cataract surgery or surgical iridectomy were visualised. Furthermore, the findings of the anterior chamber after implantation of an iris-fixated intraocular lens (IOL) and an iris prosthetic system could be assessed. The reproducibility of the anterior chamber depth measurement was +/- 22 microm (OCT) and +/- 76 microm (US). The mean difference between optical and acoustic values was 1 microm (0.03 % p = 0.921) and the limits of agreement (95 % confidence interval) were 260 microm (8.58 %). CONCLUSIONS: The anterior segment OCT proved to be a helpful diagnostic method. The cross-sectional visualisation of the entire anterior chamber allowed us to assess important values for the implantation of iris-fixated IOL and other changes after surgical procedures. The resolution and reproducibility were higher than for conventional ultrasound biometry. PMID- 16308817 TI - [Differential diagnosis and therapy of graft rejection after keratoplasty]. AB - Allograft rejection is the leading cause of corneal graft failure after perforating keratoplasty. Clinically, graft rejection may present as acute changes in corneal host tissue. All three corneal layers, epithelium, stroma and endothelium, can be affected separately or manifest combinations of rejection. Differentiation from other postoperative morphological changes following corneal transplantation can prove to be difficult, but is critical in treatment. This article is intended to provide a review on differential diagnoses and current therapeutic measures. PMID- 16308818 TI - [How "safe" is corneal transplantation? A contribution on the risk of HSV transmission due to corneal transplantation]. AB - INTRODUCTION: We have attempted in this article to summarise the present literature on the risks of virus transmission via corneal transplantation, with special emphasize on Herpes simplex virus (HSV). METHODS: We conducted a literature search from Medline. RESULTS: HSV is now acknowledged as a cause of primary graft failure, and this entity has been attributed to donor-to-host HSV infection. HSV is the primary cause of graft failure, and may induce reactivation in the recipient in the dendritic form, or in atypical forms. CONCLUSIONS: Corneal transplantation is a potential route for HSV infection as a possible transmittable disease which now has to be taken into consideration in eye banks. PMID- 16308819 TI - [Quantitative assessment of topographical parameters to differentiate keratoconus from pellucid marginal corneal degeneration and ]. AB - INTRODUCTION: The purpose is to compare videokeratometric data (Fourier series harmonic analysis and wave-front analysis) in eyes with ectatic corneal disease (keratoconus; pellucid marginal corneal degeneration [PMCD]) and to determine parameters for early diagnosis and distinction of keratoconus and PMCD. PATIENTS AND METHODS: 13 eyes with PMCD, 32 eyes with keratoconus and 18 healthy eyes were included. Computerized videokeratographic methods were assessed, using Fourier series harmonic analysis, Zernike coefficients and eccentricity. RESULTS: Irregular astigmatism (Fourier series harmonic analysis: decentration and/or irregularity) and the aberration coefficient were out of the normal range in all pathologic eyes. Both the nasal and the temporal eccentricities were greater than 0.63 in all PMCD patients, whereas either the superior or the inferior eccentricity was negative in all patients. In keraotoconic eyes, all quadrant eccentricities were positive and greater than 0.4. CONCLUSIONS: Irregular astigmatism and the aberration coefficient are useful parameters in detecting ectatic corneal disease using a computerized videokeratoscope. The calculation of the spherical equivalent (Fourier series harmonic analysis: no decrease of spherical equivalent in PMCD subjects) and of quadrant eccentricities are useful tools in distinguishing PMCD and keratoconus. PMID- 16308820 TI - [New aspects of corneal pachymetry in congenital glaucoma and pediatric aphakic glaucoma]. AB - BACKGROUND: Knowledge of individual corneal thickness in individual eyes is proving to be more and more crucial for correcting Goldmann applanation tonometry, as well as to assess its predictive value regarding the possible progression of glaucomatous damage. It has become an important factor in determining appropriate glaucoma therapy. Only few pachymetric data exist about eyes with congenital or secondary pediatric aphakic glaucoma. METHODS: 12 eyes with congenital glaucoma (6 patients, mean age 3.3 years, mean 3.9 previous operations) and 7 eyes with secondary pediatric aphakic glaucoma (4 patients, mean age 14.8 years, mean 3 previous operations) without clinical signs of corneal decompensation were investigated by ultrasound pachymetry. RESULTS: Eyes with glaucoma due to aphakia had a mean corneal thickness of 708 +/- 77.3 microm. Mean corneal thickness of the eyes with congenital glaucoma was 688.1 +/- 115.9 microm, which is far above the values known from the literature. Two eyes, which had a central corneal thickness of 646 microm and 640 microm, presented with a localised paracentral corneal thickness of only 402 microm and 405 microm, respectively, correlating with microscopic descemet scars. CONCLUSIONS: Congenital glaucoma and the pediatric aphakic glaucoma present with very heterogeneous clinical pictures. Especially for congenital glaucoma, this is confirmed by the high interindividual, but also intraindividual, variability of corneal pachymetry. PMID- 16308821 TI - [Monocentric two-year results after phakic posterior chamber lens (PRL) implantation in myopic patients]. AB - PURPOSE: Some refractive procedures such as PRK or LASIK are limited by factors, including high myopia, hyperopia, or a thin cornea. For these cases, phakic intraocular lenses have been developed. Previous models have been associated with various complications, however. In order to minimize risks involved, a new phakic posterior chamber lens (PRL, IOLTECH) has been developed. Recent data suggest a considerably lower complication rate. Our two-year results are presented. MATERIAL AND METHODS: The implanted lens (PRL) is a very thin foldable lens made of highly refractive silicone. The design allows floating of the lens and avoids contact to the natural lens. It is not fixed in the ciliary sulcus. Up to now, we have implanted this lens into 56 myopic eyes of 31 patients. Preoperative refraction was within a range of -8.5 to -21.25 D (spherical equivalent = S.E). Best corrected preoperative visual acuity was between 0.3 and 1.0. Postoperative follow-up was up to 3.5 years. In this study 44 eyes with at least 2 years follow up were included. RESULTS: Postoperative refraction was + 0.25 to - 1.0 D (SE, one patient with intended postoperative myopia). Far visual acuity without correction was 0.4 to 1.25, far visual acuity with best correction was 0.6 to 1.25. Two cataract formations were observed, one lens had to be removed due to repeated decentration. CONCLUSIONS: The PRL lens seems to be a promising choice for refractive surgery in myopic patients unfit for corneal surgery, although more results, especially long-term follow-ups are needed. PMID- 16308822 TI - [In vivo confocal microscopy in blepharitis]. AB - BACKGROUND: Dysfunction of the meibomian glands with inflammation and obstruction has been suggested to be an important factor in the pathogenesis of chronic blepharitis. Few objective tests are, however, available to examine the meibomian glands directly. PATIENTS AND METHODS: Nineteen patients with anterior blepharitis, meibomitis, meibomian gland dysfunction or severe keratoconjunctivitis sicca associated with blepharitis as well as 10 patients with normal lid margins were examined with the HRTII/RCM in vivo confocal microscope. Scans of the tear film, the tarsal conjunctiva, the hair follicles and the meibomian glands were analysed by a masked observer. RESULTS: Patients with normal lid margins exhibited a minimal round cell infiltrate in the tarsal conjunctival epithelium and largely normal ducts of the meibomian glands lined with a multilayered epithelium as well as normal gland acini. In patients with anterior blepharitis, blepharitis associated with autoimmune peripheral ulcerative keratitis and blepharitis in the context of severe dry eye, confocal microscopy disclosed normal meibomian glands. In 12 patients with blepharitis/meibomitis or meibomian gland dysfunction, profound pathology was visible with dilatation and obstruction of the meibomian gland ducts. In 15 of 19 patients with blepharitis/meibomitis, but not in meibomian gland dysfunction, an intense inflammation was observed in the tarsal conjunctival epithelium and stroma. In one patient, demodex folliculorum was evident in vivo. In patients with normal lid margins as well as in patients with blepharitis, hair follicles appeared within normal limits. CONCLUSIONS: In vivo confocal microscopy allowed the examination of the tear film, the tarsal conjunctiva, the lid margin including the lash follicles and the meibomian glands. In patients with meibomian gland disease pathological changes could be visualised and documented objectively. The presence of an inflammatory infiltrate permitted us to differentiate between meibomitis and meibomian gland dysfunction. Changes of the lash follicles do not seem to play an important role in blepharitis. Thus, in vivo confocal microscopy represents an objective technique in the classification and follow-up of patients with blepharitis. PMID- 16308823 TI - [Neurotrophic keratopathy--a post-LASIK case report]. AB - BACKGROUND: Neurotrophic keratopathy is a degenerative affection of the cornea caused by impairment of corneal sensitivity and represents a therapeutic challenge for ophthalmologists. The present article concentrates on the etiology, pathogenesis, diagnosis, therapeutic aspects and future therapeutic models for treating neurotrophic keratopathy and will present a case report: Also we evaluate the effects of autologous serum eye drops in bilateral LASIK-induced neurotrophic keratopathy with epithelial breakdown revealed by positive fluorescence and rose bengal staining and reduced tear film break-up time. METHODS: We treated a 42-year-old patient with post-LASIK neurotrophic keratopathy and tear film instability with autologous serum eye drops (5 x daily) and emulsion eye drops (Refresh Endura, Allergan, Irvine, CA, USA) after insertion of punctal plugs. RESULTS: Stabilization of vision, healing of the epithelium and reduction of the previously experienced symptoms like redness, itching and burning were achieved within 6 weeks. 10 months after changing therapy, the patient only complained about slight pain during lid movement. CONCLUSIONS: Severe denervation after bilateral LASIK disrupts ocular tear film dynamics and causes irritation symptoms of the ocular surface. Autologous serum eye drops may be an effective treatment of severe epithelial breakdown and be helpful to reestablish the disturbed ocular surface integrity, as shown by negative vital staining. Therefore, autologous serum eye drops represent a significant approach in the therapy of LASIK-induced severe dry eye and associated pain. PMID- 16308824 TI - [Significance of immediate affixation of a hard contact lens in the emergency treatment of severe alkali burns of the cornea (case report)]. AB - BACKGROUND: Severe alkali burns lead to massive limbal stem cell damage resulting in persistent epithelial defects, infiltration and stromal melting early in the disease process. A glued-on hard contact lens may serve as an "artificial epithelium" and protect the cornea from these complications. CASE REPORT: A 39 year-old male presented with severe lime burns in both eyes one week after injury. The right eye showed a totally denuded cornea and conjunctiva with circular paralimbic ischemia. In the left eye "only" two thirds of the cornea and adjacent conjunctiva and limbus were affected with less ischemia. Amniotic membrane transplantation was performed in both eyes but failed after four days already in the right eye. A hard contact lens was therefore glued on the right eye and allowed for visual acuity of 0.4 without correction in this primarily most heavily afflicted eye for 12 months duration. After removal of the contact lens, reepithelisation was quick, and function remained stable with addition of autologous serum eye drops. The primarily far less injured left eye, however, for which the amniotic membrane surgery was primarily judged to be adequate, could not be stabilised. The cornea perforated 4 weeks later necessitating a mini keratoplasty plus a glued-on contact lens, and correctable vision never became better than 1/35. CONCLUSION: The clinical course confirms former observations that a glued-on hard contact lens is an effective treatment early after alkali burn and prevents the cornea from infiltration and melting. Moreover, two aspects merit consideration: first, with proper technique and timing of the gluing-on a patient may retain useful vision throughout the period of wearing the contact lens; second, for the long period during which the cornea was sealed by the contact lens, improper repopulation by conjunctival epithelium was inhibited, and proper corneal epithelium was given a chance for repopulation after recovery of the limbal area resulting in a stable surface and useful vision without additional keratoplasty measures. PMID- 16308825 TI - Paraneoplastic vitelliform retinopathy associated with cutaneous or uveal melanoma and metastases. AB - PURPOSE: To report unusual vitelliform fundus findings in three cases of paraneoplastic retinopathy associated with metastasised cutaneous or uveal melanoma and in one case, a unique immunoreactivity response. PATIENTS AND METHODS: Observational case series. The histories of three patients with MAR-like paraneoplastic retinopathy were reviewed. Electroretinography, Goldmann perimetry, fluorescein angiography, and in one case optical coherence tomography, immunohistochemistry and Western blotting were performed. RESULTS: All patients revealed similar paraneoplastic vitelliform retinal abnormalities. Symptoms in two cases differed from the classical MAR syndrome. In one case, western blotting and immunohistochemistry demonstrated antibodies against 120-kDa, a soluble photoreceptor protein. No immunoreactivity to retinal bipolar cells was detected. CONCLUSION: The clinical, electrophysiological, and immunological findings in our patients suggest a melanoma associated paraneoplastic origin, like in MAR syndrome. However contrary to MAR syndrome, this paraneoplastic vitelliform retinopathy exhibits a peculiar fundus picture, consisting of serous macular detachment and nummular vitelliform lesions in the posterior pole. This could be an unusual presentation of MAR or a separate paraneoplastic entity. PMID- 16308826 TI - [Interferon alpha-associated retinopathy in patients with chronic hepatitis C]. AB - BACKGROUND: Interferons are a complex group of glycoproteins with antiviral, antiproliferative and immunomodulatory activity. They are used to treat different types of human neoplasms and chronic hepatitis C. Several case reports have described interferon-induced ocular side effects since the first case in 1990. PATIENT AND METHODS: We report about a 61-year-old female undergoing interferon 2alpha treatment who developed bilateral retinopathy during systemic therapy and provide a summary about ocular side effects and their consequences. CONCLUSIONS: Patients should be monitored by ophthalmologists before, during and after interferon therapy in order to avoid side effects like retinal neovascularisation and to prevent potential serious side effects. PMID- 16308827 TI - [The Vogt-Koyanagi-Harada syndrome: a rare differential diagnosis of uveitis in childhood. A case report taking into account the revised diagnostic criteria]. AB - BACKGROUND: The diagnosis of the multisystemic Vogt-Koyanagi-Harada (VKH) syndrome including ocular, neurological and dermatological manifestations is difficult due to the absence of diagnostic serological parameters and the variable onset of clinical signs and symptoms in the course of the disease. PATIENT: A 12 year old female patient was admitted to our clinic , presenting with bilateral choroiditis, vitiligo and episodes of headache. After having ruled out an underlying rheumatological or infectious disease, a VKH syndrome was suspected and effectively treated with systemic steroids. The initial diagnosis of VKH syndrome would not have been possible, if the existing diagnostic criteria had been applied. DISCUSSION: As can be seen in the case of our patient, the existing diagnostic criteria of VKH syndrome may prove to be inadequate in diagnosing VKH syndrome, particularly at the onset of the disease. Considering, that early diagnosis and treatment is crucial in improving the outcome of the disease, current diagnostic criteria have been revised. Recent retrospective data suggest a higher sensitivity for the revised diagnostic criteria. PMID- 16308828 TI - [Diabolic injury with the localization of foreign bodies in the apex of the intraorbital region ]. AB - BACKGROUND: Air-gun injuries with localisation of the pellet at the apex of the orbital cavity can cause difficult decisions in diagnostics and therapy, especially if you decide to retain the foreign body. METHODS: We report on a 14 year-old boy who was hit by an air-gun pellet. The foreign body went into the left orbit without damaging the globe or the optic nerve. RESULTS: The visual acuity was 1.0, the mobility was only slightly disturbed. A systemic therapy with antibiotics and corticosteroids could stop a series of relapses very well. CONCLUSIONS: Foreign bodies of the orbit can be retained if it is possible to deal with the surrounding reactions properly by conservative therapy. PMID- 16308829 TI - [Minimally invasive endoscopic surgery of the lacrimal drainage system--two case reports]. AB - BACKGROUND: Dacryoendoscopy is a standard procedure to directly evaluate the lacrimal system. The existence and localisation of dacryostenosis and mucosal changes as well as rare diseases in the drainage system, such as intracanalicular foreign bodies or dacryoliths, can be reliably detected and treated by minimally invasive endoscopic surgery. HISTORY AND SIGNS: A patient treated by canaliclulodacryozystorhinostomy with silicone tube intubation 5 years ago had a long symptom-free period but now shows tearing (epiphora) and occasional mucous secretion. During endoscopy residues of a silicone tube coated with a biofilm could be removed over the lacrimal canaliculus and then tested for microbiological contamination. A second patient with tearing, light mucous secretion and positive nasolacrimal duct irrigation was initially diagnosed with a tear-film disorder. Dacryoendoscopy showed a dacryolith in the saccus lacrimalis. This was consequently fragmented using a microdrill system and fully removed through the nasolacrimal duct as well as the lacrimal canaliculus. CONCLUSIONS: Endoscopy of the proximal and distal lacrimal system allows the assessment of intracanalicular structures and a reliable detection or exclusion of disorders in the draining system. This method is of great diagnostic importance as symptoms are frequently non-specific and the causes are multiple. Through endoscopy in combination with the use of a microdrill system the lacrimal system can be reopened and adequately reconstructed. PMID- 16308830 TI - [Structures of the European union--possibilities for influence by medical societies]. AB - In the EU more than 70 % of national law has already been replaced by EU law. Medical doctors are mostly not informed, how these EU-laws will pass. Many directives, regulations and decisions are important for medical doctors. A characterization of the EU- institutions is represented. PMID- 16308832 TI - Endothelial Na+-D-glucose cotransporter: no role in insulin-mediated glucose uptake. AB - A recent report indicates that the Na+-D-glucose cotransporter SGLT1 is present in capillaries of skeletal muscle and is required for insulin-mediated glucose uptake in myocytes. This result is based on the complete inhibition of insulin mediated muscle glucose uptake by phlorizin, an inhibitor of SGLT1. Using the pump-perfused rat hind limb, we measured glucose uptake, lactate efflux, and radioactive 2-deoxyglucose uptake into individual muscles with saline (control), phlorizin, insulin, and insulin plus phlorizin, as well as with saline and insulin using normal and low Na+ perfusion buffer. Insulin-mediated glucose uptake was not inhibited after correction for phlorizin interference in the glucose assay. Lactate efflux and 2-deoxyglucose uptake by individual muscles were unaffected by phlorizin. Low Na+ buffer did not affect insulin-mediated glucose uptake, lactate efflux, or 2-deoxyglucose uptake. We conclude that endothelial SGLT1 exerts no barrier for glucose delivery to myocytes. PMID- 16308833 TI - Testosterone rapidly stimulates insulin release from isolated pancreatic islets through a non-genomic dependent mechanism. AB - The action of testosterone on the 45Ca2+ uptake and insulin secretion was studied in short-term experiments using isolated pancreatic islets of Langerhans. Testosterone (1 microM) stimulated 45Ca2+ uptake within 60 seconds of incubation on similar proportion than tolbutamide. Also, the hormone rapidly increased insulin release (34%; 180 seconds) on the presence of non-stimulatory concentrations of glucose (3 mM). Impermeant testosterone-BSA significantly stimulated the secretion of insulin to a lower percentage (10%). The action of the hormone is specific--neither 17beta-E2 nor progesterone stimulated insulin secretion in the presence of 3 mM glucose. The action of testosterone on insulin secretion was dose-dependent, and at rat plasma physiological concentrations (25 nM), stimulus was 17% (p < 0.05). In conclusion, in isolated pancreatic islets experiments, physiological concentration of testosterone rapidly stimulate insulin secretion and 45Ca2+ uptake through a membrane bound mechanism. PMID- 16308834 TI - Glucagon-stimulated but not isoproterenol-stimulated glucose formation inhibition by interleukin-6 in primary cultured rat hepatocytes. AB - During prolonged sepsis, impairment of glucose supply by the liver leads to hypoglycemia. Our aim was to investigate whether proinflammatory cytokine interleukin-6, a major mediator of the hepatic acute phase reaction, could contribute to this impairment by inhibiting hepatic glucose production stimulated by glucagon or isoproterenol in rat hepatocytes. Interleukin-6 inhibited the stimulation of glucose formation from glycogen by glucagon but not by isoproterenol in cultured rat hepatocytes. This was confirmed in the perfused rat liver. In cultured hepatocytes, the increase in cyclic adenosine-3',5' monophosphate formation by glucagon was inhibited by interleukin-6, which was probably due to attenuation of glucagon binding to the glucagon receptor. The increase in cyclic adenosine-3',5'-monophosphate stimulated by isoproterenol was not affected by interleukin-6. However, the cytokine inhibited both expression of the key gluconeogenic control enzyme, phosphoenolpyruvate carboxykinase, stimulated by glucagon and isoproterenol. Thus, while increased glucose demand during the acute-phase reaction might initially be accomplished by catecholamine mediated stimulation of glucose formation from glycogen, inhibition of gluconeogenesis by interleukin-6 may contribute to the impairment of glucose homeostasis during the prolonged acute phase reaction. PMID- 16308835 TI - Short-term caloric restriction does not modify the in vivo insulin signaling pathway leading to Akt activation in skeletal muscle of Ames dwarf (Prop1(df)/Prop1(df)) mice. AB - The purpose of this study was to analyze the interaction between caloric restriction (CR) and the dwarf mutation at the level of insulin sensitivity and signal transduction. To this end, we analyzed the in vivo status of the insulin signaling system in skeletal muscle from Ames dwarf (df/df) and normal mice fed ad libitum or subjected to short-term (20-day) CR. We measured insulin-stimulated phosphorylation of the IR and IRS-1, IRS-1-p85 association and Akt activation, and the abundance of the IR, IRS-1, p85, GLUT-4 and IGF-1 receptor in skeletal muscle. In terms of glucose homeostasis, the response to CR was different in both groups of animals. In normal animals, CR induced a significant reduction in both circulating insulin and glucose levels, while CR did not modify these parameters in df/df mice. We did not find any significant alteration in either activation or abundance of signaling molecules analyzed after short-term CR in either normal or Ames dwarf mice. We conclude that the initial adaptation to CR in normal mice is an increase in insulin sensitivity without changes in insulin signal transduction, and that this adaptation is not evidenced in df/df mice, probably since they are already hypersensitive to insulin. PMID- 16308836 TI - Low levels of 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 in patients with newly diagnosed type 1 diabetes. AB - BACKGROUND AND AIMS: An epidemiological retrospective study and a recent prospective study from Finland have both concluded that vitamin D3 supplementation at birth protects individuals from type 1 diabetes later in life. Moreover, it is thought that vitamin D3 supplementation, in particular its activated form, 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3], may act as an immunomodulator, facilitating the shift from a Th1 to a Th2 immune response. The aim of this surveillance study was to measure levels of both 25-hydroxyvitamin D3 (25OHD3) and 1,25-dihydroxyvitamin D3 in patients with newly diagnosed type 1 diabetes as compared to normal subjects. METHODS: We measured plasma levels of 25 hydroxyvitamin D3 [25OHD3] and 1,25-dihydroxyvitamin D3 by radioimmunoassay in 88 consecutive patients with newly diagnosed type 1 diabetes (mean age 14.6 years; diagnosis within the last week), and in 57 healthy age and sex-matched subjects (mean age 16.5 years) born and residing in the Lazio region of continental Italy. RESULTS: Mean levels of both 25OHD3 and 1,25-(OH)2D3 were significantly lower in patients compared to controls (p < 0.01 and p < 0.03, respectively). There was no correlation between 1,25-(OH)2D3 plasma level and metabolic control status at disease diagnosis, age, gender, or most importantly, seasonality of disease diagnosis. This new observation endorses the findings of the Finnish study, even though Italy is a geographic area with more hours of sunlight than Finland. CONCLUSIONS: These findings suggest that vitamin D3 may be an important pathogenic factor in type 1 diabetes independent of geographical latitude, and that its supplementation should be considered not only at birth, but also at diagnosis of type 1 diabetes with the aim of favouring a Th2 immune response and protecting residual beta cells from further destruction. PMID- 16308837 TI - Improvement of glucose tolerance in type 2 diabetic patients: traditional vs. modern insulin regimens (results from the Austrian Biaspart Study). AB - OBJECTIVE: Major advantages of modern insulin regimens containing premixed insulin analogues in comparison to traditional insulin regimens have not been evaluated yet. The aim of the present study was to investigate whether meal related (breakfast, lunch, dinner) application of biphasic insulin aspart 30 (BIAsp 30) provides better glycaemic control than administration of biphasic human insulin 30 (BHI 30) twice per day. RESEARCH DESIGN AND METHODS: In a multi centre, randomized, open-label parallel trial, a total of 177 patients with type 2 diabetes mellitus were exposed to the two different insulin regimens described above over a study period of 24 weeks. HbA1c and glycemic exposure parameters were measured at predefined intervals. RESULTS: The mean difference between treatment groups in HbA1c after 24 weeks of treatment was 0.08% (p = 0.6419). Analysing the 7-point blood-glucose (BG) profiles, significant differences in BG levels were observed after lunch (156 vs. 176 mg/dl, p = 0.0289), before dinner (142 vs. 166 mg/dl p = 0.006) and after dinner (154 vs. 182 mg/dl p = 0.002) in favour of BIAsp 30 insulin. Prandial BG increment was lower in the BIAsp 30 group at breakfast (p = 0.057) and lunch (p < 0.0005). No difference was found regarding safety parameters in the two treatment groups. CONCLUSIONS: This study demonstrates that meal-related BIAsp 30-insulin maintains postprandial BG control more effectively than traditional BHI 30 insulin twice per day in type 2 diabetic patients. PMID- 16308838 TI - Massive insulin secretion in response to anaerobic exercise in exercise-induced hyperinsulinism. AB - Exercise-induced hyperinsulinism (EIHI) is a recently described entity characterised by recurrent episodes of hypoglycaemia induced by physical exercise. The index patient for this disorder and a matched control were subjected to aerobic and anaerobic exercise tests on a cycle ergometer. Aerobic exercise was performed at an intensity of 60% of the respective 4 mmol/l lactate threshold (40 min). Anaerobic exercise with an intensity corresponding to 130% VO2max lead to exertion within 2-3 min and elicited comparable maximal lactate levels in both subjects (10-11 mmol/l). The patient experienced a massive increase in insulin from 34 to 649 mU/l after the anaerobic test, and a lower increase in insulin from 27 to 79 mU/l during the aerobic test. Insulin concentration remained unchanged during both tests in the control. Epinephrine increased in the EIHI patient, which was probably a counterregulatory response to hypoglycaemia. The activity of lactate dehydrogenase of the index patient in isolated leukocytes as well as the response to inhibition of oxamate was normal. The hypothesis of abnormal transport or metabolism of lactate/pyruvate in the beta-cells of patients with EIHI was further supported by the parallel increase of lactate and insulin in this study elicited in particular by anaerobic exercise. PMID- 16308839 TI - The relationship between insulin sensitivity and serum adiponectin levels in three population groups. AB - Reduced plasma adiponectin levels are associated with insulin resistance. Black South Africans, like African Americans, are more insulin-resistant than BMI matched white subjects, as are Asian Indians. We investigated whether this interethnic variation in insulin resistance is due to differences in plasma adiponectin levels. Blood and anthropometric measurements were taken from black, white and Asian-Indian subjects. Serum adiponectin, lipids, glucose and insulin were measured; insulin sensitivity was calculated using HOMA. Black (HOMA = 2.62 +/- 0.99) and Asian-Indian subjects (HOMA = 3.41 +/- 2.85) were more insulin resistant than BMI-matched white (HOMA = 1.76 +/- 0.63) subjects (p = 0.0001). Furthermore, the white subjects had higher adiponectin levels (8.11 +/- 4.39 microg/ml) compared to black (5.71 +/- 2.50 microg/ml) and Asian Indian (5.86 +/- 2.50 microg/ml) subjects (p = 0.003). When all ethnic groups were combined, multiple regression analysis demonstrated that serum adiponectin levels corrected for BMI and ethnicity did not correlate with HOMA, but did explain 10.0 % of the variance in HDL-cholesterol levels. Within each ethnic group, adiponectin only correlated inversely with HOMA in white subjects. Adiponectin may play a role in determining serum HDL-cholesterol levels, but ethnic variation in insulin sensitivity is not dependent on serum levels of this adipokine. The relationship between adiponectin and insulin resistance varies across ethnic groups. PMID- 16308840 TI - Efficacy and safety of insulin glulisine in patients with type 1 diabetes. AB - Insulin glulisine (glulisine), a human insulin analogue with a rapid-acting time action profile, has been developed to fulfil the mealtime (bolus) insulin requirement in patients with diabetes. The aim of this multinational, multi centre, controlled, open-label, randomized, parallel-group study was to compare the efficacy and safety of insulin glulisine (glulisine) to that of insulin lispro (lispro) in adults diagnosed with Type 1 diabetes. Of the 683 patients randomized, 672 received treatment (339 patients received glulisine, 333 patients received lispro). Over the 26-week study, a similar reduction in mean HbA1c occurred in both groups (adjusted mean change from baseline -0.14% in both groups). The basal insulin dose was relatively unchanged from baseline in the glulisine group but increased in the lispro group (glulisine: 0.12 IU vs. lispro: 1.82 IU; p = 0.0001). As a consequence, total daily insulin dose decreased in the glulisine group but increased in the lispro group (glulisine: -0.86 IU vs. lispro: 1.01 IU; p = 0.0123). There was no relevant difference between the two groups in the reporting of symptomatic hypoglycaemia (overall, nocturnal and severe). This study demonstrates that glulisine provides equivalent glycaemic control to lispro. The clinical relevance of any difference in total daily insulin dose remains to be established. PMID- 16308841 TI - Reconsidering the sex differences in the incidence of pubertal disorders. AB - The age range of normal puberty is determined statistically; therefore, a similar percentage of boys and girls should present early or late puberty. However, far more girls present with precocious puberty and more boys with delayed puberty. We suggest that the gender differences in the incidence of pubertal disorders may be due to the following: The signs of puberty are more readily detected in girls, whereas male genitalia may not be examined during a physical examination. The description of the onset of puberty in boys is not uniform. The secular trend towards earlier pubertal maturation is more intense in girls than in boys. Therefore, there may be a fall in the age that correspond to the 3rd and 97th centiles for onset of puberty in girls while the ages at these centiles remain practically unchanged in boys. The high ratio of males to females with constitutional delay of puberty may also be attributed to the late appearance of the pubertal growth spurt in boys. The sex differences in the incidence of pubertal disorders may be due to gender dimorphism in the secular changes of pubertal maturation, to the difficulty of detecting the onset of puberty in boys and to the physiological sex differences in the timing of the pubertal growth spurt. PMID- 16308842 TI - Prolonged effect of stress (water and food deprivation) at weaning or in adult age on the triiodothyronine and histamine content of immune cells. AB - We used two days of total water and food deprivation as stress for female rats at weaning (three weeks old) and at adult age (two and a half months old). Triiodothyronine (T3) and histamine content of immune cells (lymphocytes, mast cells and monocyte-macrophage-granulocyte group in peritoneal fluid; lymphocytes, granulocytes and monocytes in blood; and lymphocytes in thymus) were studied three weeks after stress application using specific antibodies for flow cytometry and confocal microscopy. The stress at weaning increased T3 content of thymus lymphocytes. In case of adult T3, there was a cell type independent significant effect of stress, decreasing values in peritoneal fluid and slightly increasing effect in the blood. Histamine content of granulocytes was also significantly elevated. The experiments demonstrate that not only fetal or neonatal stress has long-lasting consequences, but also stress events in later periods of life in cells (organs) that are continuously differentiating. We will go on to discuss the importance of T3 and histamine in connection with stress and immunity. PMID- 16308844 TI - Automated optimization of mass spectrometric parameters in 'Tune' mode for impure analytes in dirty samples using gradient high-performance liquid chromatography. PMID- 16308845 TI - Detection limits of matrix-assisted laser desorption/ionisation mass spectrometry coupled to chromatography--a new application of solvent-free sample preparation. AB - The detection limits of matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS) by semi-online coupling with chromatography were investigated using various mixtures of polyethylene oxides (PEOs) with different end groups. In contrast to the common 'dried-droplet' sample preparation technique, which results in an inhomogeneous sample-to-matrix ratio within the MALDI spot, the used coupling technique offers a very high reproducibility combined with surpassing sensitivity of a few femtograms over a broad range of sample-to-matrix ratios. These results are in correlation with the results of the recently established solvent-free MALDI-TOFMS method utilising the grinding approach and are also of assistance towards the more theoretical aspect of MALDI that suggests that there is no necessity for an analyte incorporation into a matrix crystal for excellent matrix-assistance. PMID- 16308846 TI - Two-dimensional correlation spectroscopy techniques applied to ion trap tandem mass spectrometric analysis: nitroaromatics. AB - The technique of two-dimensional (2D) correlation spectroscopy is combined with ion trap tandem mass spectrometric (MS/MS) analysis to provide insight into the energetics and behavior of gas-phase ions. The 2D correlation method is demonstrated by the analysis of the fragment ions produced by the fragmentation of molecular radical cations of nitrobenzene and m-nitrotoluene at a series of applied potentials that effect collision-induced dissociation (CID). The asynchronous correlation intensities show that competitive loss of NO to form [M NO]+ proceeds at a lower CID energy than loss of NO2 to form [M-NO2]+. The formation of the ion corresponding to [M-NO-CO]+ is shown by the method to occur at a lower CID energy than the formation of [M-NO2]+, but at a higher CID energy than [M-NO]+ for both nitrobenzene and m-nitrotoluene. PMID- 16308847 TI - Identification of a trace colored impurity in drug substance by preparative liquid chromatography and mass spectrometry. AB - 6-(Nitrooxy)hexyl-(2z)-4-(acetyloxy)-3-[4-(methylsulfonyl)phenyl]-2-phenylbut-2 enoate (enoate 1) was investigated as a novel therapy for pain relief. In a recent manufacturing run at the pilot plant scale, the enoate drug substance was found to have a yellowish color not observed previously. An unknown impurity at trace level was detected by high-performance liquid chromatographic (HPLC) analysis and found to be the primary cause for the color of the drug substance. The colored impurity was enriched by preparative HPLC and structurally elucidated by liquid chromatography/tandem mass spectrometry (LC/MS/MS). It was found that the colored impurity was derived from the product of oxidative dimerization of rofecoxib, an impurity present in the enoic acid intermediate. It was further revealed by the photodiode array and LC/MS/MS data that the colored impurity exists in the drug substance as a pair of double-bond isomers with one isomer at majority. These findings were also confirmed by synthesizing the colored impurity through the proposed pathway. PMID- 16308848 TI - The formation of neutral CCCO2H and HCCCO2 molecules from anionic precursors in the gas phase: a joint experimental and theoretical study. AB - Calculations at the CCSD(T)/aug-cc-pVDZ//B3LYP/6-31G(d) level of theory indicate that the anions -CCCO2H and HCCCO2(-) are stable species in their singlet states. Upon collision-induced, vertical one-electron oxidation under neutralisation reionisation (-NR+) conditions, they produce the neutral molecules CCCO2H and HCCCO2, respectively. Some of the CCCO2H neutrals should be stable for the duration of the neutralisation-reionisation experiment (10(-6) s), while others will dissociate to CCCO and OH (requires 125 kJ mol(-1)). In contrast, neutral HCCCO2 is expected to be much less stable, and dissociate to HCC and CO2 (37 kJ mol(-1)). Neither CCCO2H nor HCCCO2 is expected to interconvert, or to rearrange to other isomers. The anions -CCCO2H and HCCCO2(-) have been formed in the ion source of the mass spectrometer by the reactions between (CH3)3Si-C[triple bond]C CO2H and F- and HC[triple bond]C-CO2Si(CH3)3 and F-, respectively. The -NR+ spectrum of -CCCO2H shows a recovery signal and also indicates that the lowest energy dissociation pathway of neutral CCCO2H corresponds to the loss of OH. The NR+ spectrum of HCCCO2 displays little or no recovery signal, and the spectrum is dominated by the [CO2]+ ion. The experimental observations are in agreement with the predictions of the extensive theoretical studies. PMID- 16308849 TI - Blood vessel stem cells and wound healing. AB - This article continues the Journal's "Scientific Surgery" series of leaders. The aim of the series, published throughout 2005, has been to highlight areas of bioscience that, while still largely confined to the experimental laboratory, may soon be brought into the clinical domain. In this month's paper Watt and Fox offer an up to date insight into the processes of tissue healing and suggest possible future therapeutic strategies. PMID- 16308850 TI - Study of fragmentation pattern and adsorption of 9-O-(triphenylsilyl)-10,11 dihydrocinchonidine on platinum by hydrogen/deuterium exchange using electrospray ionization ion-trap tandem mass spectrometry. AB - We have studied the adsorption on a platinum (Pt) catalyst of two compounds utilizable as a chiral basic catalyst and a chiral modifier, dihydrocinchonidine (DHCD), and a new cinchona alkaloid derivative containing a bulky group, the Ph3SiO-DHCD molecule. The method of choice was the detection by electrospray ionization (ESI) ion-trap tandem mass spectrometry (MS/MS) of hydrogen/deuterium (H/D) exchange at room temperature, in tetrahydrofuran, at a D2 pressure of 1 bar. Based on the ESI-MS/MS spectrum of the new compound, we propose a mechanism for the formation of the silatropylium cation containing a Si-O bond. From the fragmentation pattern of Ph3SiO-DHCD it was confirmed that ESI-ion-trap MS/MS can be used to study the adsorption processes of complicated carbon compounds by investigating their H/D exchange reactions. In the case of Ph3SiO-DHCD, the results demonstrate that H/D exchange takes place mainly on the quinoline skeleton. However, the strong pi-bonded adsorption of the quinoline skeleton parallel with the imaginary plane of Pt is not preferred because the bulky Ph3Si group inhibits the multiple pi-bonded adsorption of the Ph3SiO-DHCD. Because of this hindrance the molecule was adsorbed tilted via the nonbonding electron pair of the N atom and C2' atom of the quinoline skeleton; consequently, mainly alkaloid-d1 and alkaloid-d2 are formed. PMID- 16308851 TI - Proteomics and clinical surgery. AB - This article concludes the Journal's "Scientific Surgery" series of leaders. The series, published throughout 2005, has highlighted areas of bioscience that may soon be transferred from the laboratory into the clinical sphere. In this final paper Roblick and Auer consider the future role of the surgeon in the era of the "diagnostic chip". PMID- 16308852 TI - High precision measurements of 17O/16O and 18O/16O ratios in H2O. AB - We have optimized the method of water fluorination using the solid reagent CoF3 to produce O2. This allows isotope ratio measurements by dual-inlet mass spectrometry with very high precision of 0.01 to 0.03/1000 for both delta17O and delta18O. Using this method, delta17O and delta18O of atmospheric O2 were determined as 12.08 and 23.88/1000 vs. VSMOW, respectively. Likewise, delta17O and delta18O of GISP were -13.12 and -24.73/1000, and for SLAP they were -29.48 and -55.11/1000 vs. VSMOW, respectively. Analysis of these data in a ln(delta17O + 1) vs. ln(delta18O + 1) plot yields a line with a regression coefficient (lambda) of 0.5279 +/- 0.0001 (R2 = 0.999999). We also determined the fractionation factors 17alpha and 18alpha in liquid-vapor equilibrium, and found that the ratio ln 17alpha/ln 18alpha is constant (0.529 +/- 0.001) over the temperature range 11.4 to 41.5 degrees C. PMID- 16308853 TI - Randomized clinical trial comparing blue dye with combined dye and isotope for sentinel lymph node biopsy in breast cancer. AB - BACKGROUND: Use of blue dye alone as a marker for sentinel lymph node (SLN) biopsy is effective, but combining it with isotope marking can improve the success rate. Use of the isotope adds extra cost and there are potential radiation hazards. The two techniques were compared in a randomized trial. METHODS: Women with early breast cancer (less than 3 cm) and no palpable axillary nodes were recruited. Women older than 70 years with multicentric cancers or previous surgery to the breast or axilla were excluded. Patients were randomized to either blue dye alone or combined mapping for SLN biopsy. All women had a level I and II axillary dissection after the SLN biopsy. RESULTS: A total of 123 patients were recruited, of whom five were excluded from analysis. Blue dye alone was used in 57 women and 61 had combined mapping. Baseline demographic data were similar in the two cohorts. The success rate of SLN biopsy was higher with combined mapping than with blue dye alone (100 versus 86 per cent; P = 0.002). The accuracy and false-negative rate were similar (accuracy 100 per cent for combined mapping versus 98 per cent for blue dye; false-negative rate 0 versus 5 per cent). CONCLUSION: Combined mapping was superior to blue dye alone in identification of the SLN, but accuracy and false-negative rates were similar. PMID- 16308855 TI - Randomized clinical trial comparing lightweight composite mesh with polyester or polypropylene mesh for incisional hernia repair. AB - BACKGROUND: Polymer mesh has been used to repair incisional hernias with lower recurrence rates than suture repair. A new generation of mesh has been developed with reduced polypropylene mass and increased pore size. The aim of this study was to compare standard mesh with new lightweight mesh in patients undergoing incisional hernia repair. METHODS: Patients were randomized to receive lightweight composite mesh, or standard polyester or polypropylene mesh. Outcomes were evaluated at 21 days, 4, 12 and 24 months from patient responses to the Short Form 36 (SF-36) and daily activity questionnaires. Complications and recurrence rates were recorded. RESULTS: A total of 165 patients were included in an intention-to-treat analysis (83 lightweight mesh, 82 standard mesh). Postoperative complication rates were similar. The overall hernia recurrence rate was 17 per cent with the lightweight mesh versus 7 per cent with the standard mesh (P = 0.052). There were no differences in SF-36 physical function scores or daily activities between 21 days and 24 months after surgery. CONCLUSION: The use of the lightweight composite mesh for incisional hernia repair had similar outcomes to polypropylene or polyester mesh with the exception of a non significant trend towards increased hernia recurrence. The latter may be related to technical factors with regard to the specific placement and fixation requirements of lightweight composite mesh. PMID- 16308856 TI - Systematic review of dressings and topical agents for surgical wounds healing by secondary intention (Br J Surg 2005; 92: 665-672). PMID- 16308857 TI - A cornucopia of vertebrate brains: new insights into nature's diversity and how it came to be. PMID- 16308858 TI - Regenerative medicine and developmental biology: the role of the extracellular matrix. AB - The principles and ultimate goals of regenerative medicine and developmental biology involve a complex sequence of events, culminating in the formation of structurally and functionally normal tissues and organs. The molecular composition of the extracellular matrix (ECM) plays a critical role in cellular migration and differentiation events. Mammalian ECM, derived from various tissues and organs, has been used as a biologic scaffold for therapeutic regenerative applications. Hundreds of thousands of human patients have benefited from the use of biologic scaffolds composed of naturally occurring ECM. The mechanisms by which ECM induces constructive remodeling instead of scar tissue formation are only beginning to be understood. This article reviews composition of mammalian ECM, its poorly understood role in developmental biology, and the clinical applications that have resulted from the use of this naturally occurring scaffold. PMID- 16308859 TI - Some principles of regeneration in mammalian systems. AB - This article presents some general principles underlying regenerative phenomena in vertebrates, starting with the epimorphic regeneration of the amphibian limb and continuing with tissue and organ regeneration in mammals. Epimorphic regeneration following limb amputation involves wound healing, followed shortly by a phase of dedifferentiation that leads to the formation of a regeneration blastema. Up to the point of blastema formation, dedifferentiation is guided by unique regenerative pathways, but the overall developmental controls underlying limb formation from the blastema generally recapitulate those of embryonic limb development. Damaged mammalian tissues do not form a blastema. At the cellular level, differentiation follows a pattern close to that seen in the embryo, but at the level of the tissue and organ, regeneration is strongly influenced by conditions inherent in the local environment. In some mammalian systems, such as the liver, parenchymal cells contribute progeny to the regenerate. In others, e.g., skeletal muscle and bone, tissue-specific progenitor cells constitute the main source of regenerating cells. The substrate on which regeneration occurs plays a very important role in determining the course of regeneration. Epimorphic regeneration usually produces an exact replica of the structure that was lost, but in mammalian tissue regeneration the form of the regenerate is largely determined by the mechanical environment acting on the regenerating tissue, and it is normally an imperfect replica of the original. In organ hypertophy, such as that occurring after hepatic resection, the remaining liver mass enlarges, but there is no attempt to restore the original form. PMID- 16308860 TI - Limb regeneration in higher vertebrates: developing a roadmap. AB - We review what is known about amphibian limb regeneration from the prospective of developing strategies for the induction of regeneration in adult mammals. Prominent in urodele amphibian limb regeneration is the formation of a blastema of undifferentiated cells that goes on to reform the limb. The blastema shares many properties with the developing limb bud; thus, the outgrowth phase of regeneration can be thought of as cells going through development again, i.e., redevelopment. Getting to a redevelopment phase in mammals would be a major breakthrough given our extensive understanding of limb development. The formation of the blastema itself represents a transition phase in which limb cells respond to injury by dedifferentiating to become embryonic limb progenitor cells that can undergo redevelopment. During this phase, rapid wound closure is followed by the dedifferentiation of limb cells to form the blastema. Thus, the regeneration process can be divided into a wound-healing/dedifferentiation phase and a redevelopment phase, and we propose that the interface between the wound-healing response and gaining access to developmentally regulated programs (dedifferentiation) lies at the heart of the regeneration problem in mammals. In urodele amphibians, dedifferentiation can occur in all of the tissues of the limb; however, numerous studies lead us to focus on the epidermis, the dermis, and muscle as key regulators of regeneration. Among higher vertebrates, the digit tip in mammals, including humans, is regeneration-competent and offers a unique mammalian model for regeneration. Recent genetic studies in mice identify the Msx1 gene as playing a critical role in the injury response leading to digit tip regeneration. The results from regeneration studies ranging from amphibians to mammals can be integrated to develop a roadmap for mammalian regeneration that has as its focus understanding the phenomenon of dedifferentiation. PMID- 16308861 TI - Cellular plasticity in vertebrate regeneration. AB - Within the animal kingdom, there are several examples of organisms with remarkable regenerative abilities. Among vertebrates, newts appear to be the most adept at replacing lost structures and injured organs and can regenerate their limbs, tails, spinal cords, jaws, retinas, lenses, optic nerves, intestines, and heart ventricles. This regenerative ability is dependent on the induction of an unusual degree of cellular plasticity near the site of injury. Mature cells lose their differentiated characteristics and revert to proliferating progenitor cells that will later redifferentiate to replace the lost or injured tissues. This degree of cellular plasticity appears to be restricted to those vertebrates with the most remarkable regenerative abilities and is not observed in mammals. However, in the last several years, there have been a few studies suggesting that certain factors present in newt tissues can induce a dedifferentiation response in mammalian cells. These results suggest that the knowledge gained from studying the molecular basis of cellular plasticity in newts and other regeneration competent model organisms might one day be used to enhance the regenerative potential in mammals. PMID- 16308862 TI - Eye on regeneration. AB - Lens regeneration in newts is a remarkable process, whereby a lost tissue is replaced by transdifferentiation of adult tissues that only a few organisms possess. In this review, we will touch on the approaches being used to study this phenomenon, recent advances in the field of lens regeneration, similarities and differences between development and regeneration, as well as the potential role stem cells may play in understanding this process. PMID- 16308865 TI - Protective defunctioning stoma in low anterior resection for rectal carcinoma (Br J Surg 2005; 92: 1137-1142). PMID- 16308866 TI - Preoperative plasma N-terminal pro-brain natriuretic peptide as a marker of cardiac risk in patients undergoing elective non-cardiac surgery (Br J Surg 2005; 92: 1041-1045). PMID- 16308867 TI - Physical, chemical and environmental properties of selected chemical alternatives for the pre-plant use of methyl bromide as soil fumigant. AB - Production and use of methyl bromide, a soil fumigant, are being restricted because of this chemical's deleterious effects on stratospheric ozone concentrations. Several products, some of which are currently used as soil fumigants, are being considered as possible replacements for methyl bromide, alone and in various combinations. Among these, 1,3-dichloropropene, methyl isothiocyanate generators such as metam-sodium, and chloropicrin are currently registered, while others such as methyl iodide and sodium azide are at different stages of the registration process. This review examines physicochemical properties, environmental fate, and metabolism of the various potential methyl bromide replacement products. PMID- 16308868 TI - Selectivity of pesticides used on cotton (Gossypium hirsutum) to Trichogramma pretiosum reared on two laboratory-reared hosts. AB - The side-effects of pesticides (insecticides, fungicides, herbicides and plant growth regulators) used on cotton were tested on adults and pupae of Trichogramma pretiosum Riley reared in the laboratory on two different hosts, the Angoumois grain moth (Sitotroga cerealella Olivier) and the Mediterranean flour moth (Ephestia kuehniella (Zeller)). The eggs of the host enclosing the parasitoid pupae received direct pesticide sprays, while the adults of the parasitoid were exposed to the pesticides through contact with residues on sprayed eggs offered to parasitism. Alpha-cypermethrin, carbosulfan, deltamethrin, endosulfan, profenofos and zeta-cypermethrin were highly noxious to the parasitoid, significantly reducing the percentage of emergence and parasitism of T. pretiosum developing in E. kuehniella or S. cerealella eggs. However, the pupal stage of the parasitoid developing in S. cerealella eggs was less susceptible to alpha cypermethrin and deltamethrin. Lufenuron and metamidophos greatly reduced the percentage of adult emergence from eggs of both hosts, while novaluron only interfered on this variable when the wasps were developing in E. kuehniella eggs. However, lufenuron and monocrotophos had no effect on the parasitoid pupae of T. pretiosum developing in E. kuehniella eggs. Chlorfluazuron, diafenthiuron, diflubenzuron, fentin hydroxide, mepiquat chloride, novaluron, thiacloprid and triflumuron did not affect T. pretiosum emergence when eggs of S. cerealella enclosing pupae of the wasps were surface treated. The pesticides azoxystrobin, carbendazin + thiram, mepiquat chloride and novaluron had no effect on the ability of the wasps to parasitise E. kuehniella eggs. However, only mepiquat chloride did not affect the percentage of F1 wasps emerging from E. kuehniella eggs. The remaining pesticides moderately reduced the percentage of emergence and parasitism of the wasps when they had contact with the chemicals during their pupal or adult stage. Thus there were differences in pesticide toxicity according to the host used for parasitoid development. These differences were hypothesised to occur because of differences in egg morphology and parasitoid performance. PMID- 16308869 TI - Modelling the effects of alcohol ethoxylates on diffusion of pesticides in the cuticular wax of Chenopodium album leaves. AB - Cuticular waxes represent the first and, in most cases, the limiting barrier for foliar uptake of pesticides from solution. Sorption of pesticides in reconstituted cuticular wax (wax/water partition coefficients) of Chenopodium album L. and in isolated cuticular membranes (cuticle/water partition coefficients) of Prunus laurocerasus L. was determined. Diffusion coefficients of pesticides in reconstituted cuticular wax of C. album leaves were size-dependent, increasing with increasing molar volume. In the presence of alcohol ethoxylates, diffusion coefficients were enhanced by up to two orders of magnitude, and size selectivity was significantly decreased. The accelerating effect and the decrease in size selectivity were attributed to plasticisation of the cuticular wax by the alcohol ethoxylates increasing the fluidity in the wax. A free volume model adopted from polymer science was successfully applied to predict diffusion coefficients of pesticides on the basis of the transport properties of the wax (size selectivity and crystallinity), the molar volume of the diffusing compound and the accelerator concentration in the wax. PMID- 16308870 TI - Rearrangement of chromosomal region 8q11-13 in lipomatous tumours: correlation with lipoblastoma morphology. AB - Cytogenetics is of considerable value when diagnosing lipomatous tumours, as different tumour types have different more or less specific chromosomal abnormalities. One such entity is lipoblastoma, which is a benign lipomatous tumour that often exhibits rearrangements of chromosome bands 8q11-13, and the gene PLAG1 has been implicated as the target of these chromosomal changes. All lipomatous tumours karyotyped at the Norwegian Radium Hospital were reviewed, looking for rearrangements of 8q11-13. Five tumours exhibiting chromosomal abnormalities affecting this region were found. Only one of them was morphologically diagnosed as a lipoblastoma, two being classified as lipomas, one as a hibernoma, and one as a well-differentiated liposarcoma. The two tumours successfully analysed with bacterial artificial chromosomes (BACs) covering the gene PLAG1 showed involvement of this gene in the rearrangement. The findings raise the question as to what extent the diagnosis lipoblastoma should be based on histopathological or cytogenetic/molecular data or a combination thereof. When karyotypic information from this series was combined with available literature data, it was found that the sensitivity of 8q11-13 rearrangements for diagnosing lipoblastomas when found in a lipomatous tumour was 77% and that the corresponding specificity was 98%. The validity of these calculations of the diagnostic information provided by the cytogenetic findings is, of course, totally dependent on the morphological diagnosis made in each case. Regardless of what the precise phenotypic diagnosis was, it is suggested that lipomatous tumours with 8q11-13 rearrangement constitute a distinct pathogenetic entity. When selective therapies tailor-made against the specific pathogenetic rearrangement become available, it will become mandatory to pay more attention to the genetic constitution of the tumour cells than to their phenotypic appearance. PMID- 16308871 TI - Long-term persistence and efficacy of spinosad against Rhyzopertha dominica (Coleoptera: Bostrychidae) in wheat. AB - A laboratory study was undertaken to determine the persistence and efficacy of spinosad against Rhyzopertha dominica (F.) in wheat stored for 9 months at 30 degrees C and 55 and 70% relative humidity. The aim was to investigate the potential of spinosad for protecting wheat from R. dominica during long-term storage in warm climates. Wheat was treated with spinosad at 0.1, 0.5 and 1 mg kg(-1) grain and sampled after 0, 1.5, 3, 4.5, 6, 7.5 and 9 months of storage for bioassays and residue analyses. Residues were estimated to have declined by 30% during 9 months of storage at 30 degrees C and there was no effect of relative humidity. Spinosad applied at 0.5 or 1 mg kg(-1) was completely effective for 9 months, with 100% adult mortality after 14 days of exposure and no live F1 adults produced. Adult mortality was <100% in some samples of wheat treated with 0.1 mg kg(-1) of spinosad, and live progeny were produced in all samples treated at this level. The results show that spinosad is likely to be an effective grain protectant against R. dominica in wheat stored in warm climates. PMID- 16308872 TI - Improved 2-nitrobenzenesulfenyl method: optimization of the protocol and improved enrichment for labeled peptides. AB - We have developed the NBS (2-nitrobenzenesulfenyl) method, a quantitative proteome analysis method utilizing stable isotope labeling followed by mass spectrometry. The potential of this method was reported previously, and the procedure has now been further optimized. Here, we describe a procedure utilizing urea or guanidine hydrochloride as a protein denaturant, in conjunction with an improved chromatographic enrichment method for the NBS-labeled peptides using a phenyl resin column. By using this new protocol, both sample loss throughout the protocol and the elution of unwanted unlabeled peptides can be minimized, improving the efficiency of the analysis significantly. PMID- 16308873 TI - Rapid and direct measurement of free concentrations of highly protein-bound fluoxetine and its metabolite norfluoxetine in plasma. AB - Fluoxetine (F) and its active N-demethylated metabolite, norfluoxetine (NF), are selective serotonin re-uptake inhibitors that bind extensively to plasma proteins. Development and validation of a novel method for measuring free concentrations of F and NF in plasma are reported here. The plasma filtrate was prepared by a high-speed short-duration ultrafiltration (UF) and then submitted directly to a short-column liquid chromatography/tandem mass spectrometric (LC/MS/MS) assay. There was no significant matrix effect on the analysis, and non specific binding of the analytes to the UF devices was negligible. For validation of the method, the recovery of the free analytes was compared to that from an optimized equilibrium dialysis method, and analyte stability was examined under conditions mimicking the sample storage, handling, and analysis procedures. The linearity range was 0.37-12 ng/mL for F and NF; the within-run and between-run relative standard deviations were less than 11.9%, and accuracies across the assay range were 100 +/- 10.3%. This new method was then further validated in a pharmacokinetic (PK) study in beagle dogs receiving a single oral dose of fluoxetine hydrochloride. The integrity of the resulting PK data of free F and NF was absolute. The PK data indicate that the novel method is accurate and reliable. To our knowledge this is the first report describing a rapid and reliable method for direct measurement of free concentrations of F and NF in plasma, which will be useful for clinical pharmacokinetic/pharmacodynamic studies of F. Furthermore, the strategies described herein may be applied to the development and validation of methods for measuring the free concentrations of other drugs in plasma. PMID- 16308874 TI - Top-down characterization of proteins and drug-protein complexes using nanoelectrospray tandem mass spectrometry. AB - We report a 'top-down' approach for characterization of proteins, and identification of binding sites in protein-drug complexes using nanoelectrospray ionization hybrid quadrupole time-of-flight tandem mass spectrometry (nanoESI MS/MS). The efficiency of direct fragmentation of intact protein ions and the feasibility of this method were initially demonstrated using several well characterized proteins with different molecular weights including metallothionein (6126 Da), cytochrome c (horse, 12360 Da), myoglobin (horse, 16592 Da), and hemoglobin (human, 64453 Da). Simply varying collision energy without enzyme digestion and gel or LC separation generated a range of peptide fragments of these proteins. Over 80% of these peptide ions matched those in the SWISS-PROT database with mass accuracy of 8 to 32 ppm with external calibration. This technique was further applied to fragment a cisplatin-metallothionein complex to identify the binding sites, demonstrating a potential application in the study of drug-protein binding. PMID- 16308875 TI - Characterization of boldione and its metabolites in human urine by liquid chromatography/electrospray ionization mass spectrometry and gas chromatography/mass spectrometry. AB - Boldione (1,4-androstadiene-3,17-dione) is a direct precursor (prohormone) to the anabolic steroid boldenone (1,4-androstadiene-17beta-ol-3-one). It is advertised as a highly anabolic/androgenic compound promoting muscularity, enhancing strength and overall physical performance, and is available on the Internet and in health stores. This work was undertaken to determine and characterize boldione and its metabolites in human urine, using both liquid chromatography with electrospray ionization mass spectrometry and gas chromatography with mass spectrometry and derivatization. Boldione and its three metabolites were detected in dosed human urine after dosing a healthy volunteer with 100 mg boldione. The excretion studies showed that boldione and its metabolites were detectable in urine for 48 h after oral administration, with maximum excretion rates after 1.8 and 3.6 h (boldenone case). The amounts of boldione and boldenone excreted in urine from this 100 mg dose were 34.45 and 15.95 mg, respectively. PMID- 16308877 TI - Direct infusion electrospray ionization mass spectra of crude cell extracts for microbial characterizations: influence of solvent conditions on the detection of proteins. AB - Direct infusion electrospray ionization mass spectrometry (DIES-MS) of crude bacterial extracts is a rapid method that can be used to characterize microbial cells. Phospholipids, metabolites, and proteins can be detected rapidly with minimal sample preparation. However, several factors influence the detection of signals in such high-throughput analyses. We studied the influence of solvent conditions, including the organic content and pH of the solvent, on the extraction and subsequent detection of signals in DIES-MS, with a view to improving the detection of protein signals. Unfractionated cell extracts from three strains of the Gram-negative Escherichia coli (including one encoding a recombinant green fluorescence protein), and the Gram-positive Bacillus sphaericus and B. subtilis were investigated. Both pH and the organic content of the solvent were found to influence the spectral information as observed from principal component analysis of the spectral data. A polar solvent with higher organic content resulted in the extraction of phospholipids that overtly dominate the spectral information. Decreasing the organic content of the extraction solvent resulted in the improved detection of protein peaks. Altering the pH of the extraction solvent resulted in different protein profiles from the same bacterium, as observed after spectral deconvolution. In addition, the protein profiles were also different when using different organic solvents. Spectral deconvolution showed several protein peaks that had mass-based homology with those in protein databases for the (sequenced) organisms studied. These results suggest that a combination of solvent conditions can be used to generate protein profiles rapidly that when combined can provide additional valuable proteomic information. PMID- 16308878 TI - Electrodeposition of metals and semiconductors in air- and water-stable ionic liquids. AB - In addition to their stability, the advantages of air- and water-stable ionic liquids over chloroaluminate ionic liquids, which were intensively investigated in the past, are that they are easy to dry, purify, and handle. Moreover, some of these ionic liquids have an extremely large electrochemical window of more than 5 V, and hence they give access to the electrodeposition of many metals and semiconductors, such as Ta, Ti, Si, and Ge. The results to date for the electrodeposition of metals and semiconductors in the most popular air- and water stable ionic liquids are presented. PMID- 16308879 TI - Hollow silica spheres with an ordered pore structure and their application in controlled release studies. AB - Herein we report the synthesis and characterization of hollow silica spheres with a narrow size distribution, uniform wall thickness, and a worm-like pore structure. The formation of these spheres was monitored by confocal laser scanning microscopy and dynamic light scattering. A model for the molecular build up of these silica hollow spheres is derived from these data in combination with studies of the as-made particles by transmission electron microscopy, scanning electron microscopy, pore size analysis, thermogravimetric analysis, and solid state nuclear magnetic resonance. We further demonstrate that these spheres can be used for the encapsulation and subsequent release of different dye molecules. PMID- 16308880 TI - Synthesis and photophysical properties of annulated dinuclear and trinuclear phthalocyanines. AB - Metal-free mononuclear, dinuclear and trinuclear phthalocyanines were prepared by a mixed cyclotetramerisation of a 1,2,4,5-tetracyanobenzene derivative and 4,5 bis(2,6-dimethylphenoxy)phthalonitrile. For the first time, a pi-electron conjugated trinuclear phthalocyanine was synthesised with phthalocyanine units connected by common annulated benzene rings. The Q band of the trinuclear compound in solution occurs at lambda = 944 nm whereas those of the dinuclear and mononuclear compounds are at lambda = 853/830 and 701/664 nm, respectively. Fluorescence quantum yields, fluorescence lifetimes and singlet-oxygen quantum yields of the compounds were determined. PMID- 16308881 TI - A simple and efficient approach to 1,3-polyols: application to the synthesis of cryptocarya diacetate. AB - A highly enantio- and stereoselective synthetic strategy for both syn- and anti 1,3-polyols has been developed. The sequence involves iterative Jacobsen's hydrolytic kinetic resolution (HKR), diastereoselective iodine-induced electrophilic cyclization, and ring-closing metathesis (RCM). This protocol has subsequently been utilized for the synthesis of cryptocarya diacetate, a natural product with broad range of biological activity. PMID- 16308882 TI - Proteo-dendrimers designed for complementary recognition of cytochrome c: dendrimer architecture toward nanoscale protein complexation. AB - "Proteo-dendrimers" in which polyanionic hepta(glutamic acids), fluorescent zinc porphyrinate cores, hydrophilic polyether surfaces, and nonpeptide hydrophobic dendrons are combined, were developed as a new series of synthetic receptors for protein recognition. They have polyanionic "patch" structures on their surfaces and undergo complementary electrostatic interactions with a positively charged cytochrome c patch, as observed in biological protein-protein recognition systems. Stability constants of the resulting supramolecular complexes were determined in phosphate buffer (pH 7) by monitoring the fluorescence quenching of the zinc porphyrinates. These proteo-dendrimer receptors exhibited higher affinities with cytochrome c proteins in aqueous solutions than with biological cytochrome b5. Furthermore, they effectively blocked complexation of biological cytochrome b5 with cytochrome c, indicating that the proteo-dendrimers and cytochrome b5 similarly occupy the polycationic patch of cytochrome c. PMID- 16308883 TI - Fetoscopic and ultrasound-guided decompression of the fetal trachea in a human fetus with Fraser syndrome and congenital high airway obstruction syndrome (CHAOS) from laryngeal atresia. AB - Congenital high airway obstruction syndrome (CHAOS) from laryngeal atresia bears a poor prognosis for hydropic fetuses owing to cardiac failure. We attempted percutaneous fetoscopic and ultrasound-guided tracheal decompression in a hydropic human fetus with CHAOS associated with Fraser syndrome. Percutaneous fetoscopic and ultrasound-guided tracheal decompression was performed using three trocars under general materno-fetal anesthesia at 19 + 5 weeks of gestation. Abnormal fetoplacental blood flow normalized within hours as a result of the intervention. Furthermore, a normalization of lung : heart size and lung echogenicity was observed within days. Resolution of hydrops was complete within 3 weeks. Premature rupture of membranes and premature contractions prompted emergency delivery of the fetus by ex-utero intrapartum treatment (EXIT) at 28 + 2 weeks of gestation. Following delivery, the lungs could be ventilated at low pressures and ambient oxygen concentration. Weaning from ventilation was achieved at 18 days of postnatal life. Our experience indicated that percutaneous fetoscopic and ultrasound-guided decompression of the fetal trachea is feasible and may permit normalization of hemodynamics in hydropic human fetuses with CHAOS from laryngeal atresia. The procedure may also result in normalization of heart : lung size and provide the time needed to regain the function of the overstretched diaphragm in this grave fetal condition. PMID- 16308884 TI - Clinical evaluation of plasma free phenytoin measurement and factors influencing its protein binding. AB - The relationship between free phenytoin concentrations and clinical responses, and the factors influencing protein binding of phenytoin were investigated. A total of 119 plasma samples from 70 patients treated orally with phenytoin were analysed. The mean free phenytoin concentration was significantly higher in the patients who received phenytoin monotherapy and were classified as having a complete response (1.25 +/- 1.09 microg/ml) than that in the partial response group (0.59 +/- 0.07 microg/ml), whereas the mean total concentrations were not significantly different between the two groups. Samples were divided into three groups based on the free fraction of phenytoin, i.e. low, <5%; medium, 5%-10%; high, > 10%. The mean age (55.3 +/- 10.9 years) was significantly higher in the high group than in the low (42.7 +/- 21.2 years) and medium (42.8 +/- 16.0 years) groups. The mean creatinine clearance (CLcr) (55.3 +/- 10.9 ml/min) and serum albumin concentration (3.30 +/- 1.25 g/dl) were significantly lower in the high group than the low (88.3 +/- 29.0 ml/min and 4.08 +/- 0.50 g/dl, respectively) and medium (95.0 +/- 32.8 ml/min and 3.95 +/- 0.92 g/dl, respectively) groups. These results suggest that the free phenytoin concentration, rather than the total concentration, is more useful for monitoring antiepileptic effects in patients receiving phenytoin monotherapy. It was also found that the free phenytoin fraction was significantly influenced by aging, CLcr and serum albumin levels. PMID- 16308885 TI - A single immunization with a minute dose of a lentiviral vector-based vaccine is highly effective at eliciting protective humoral immunity against West Nile virus. AB - BACKGROUND: Lentiviral vectors, due to their capacity to transduce non-dividing cells, have become precious and worldwide used gene transfer systems. Their ability to efficiently and stably transduce dendritic cells (DCs) has led to their successful use as vaccination vectors for eliciting strong, specific and protective cellular immune responses mostly in anti-tumoral but also in anti viral applications. However, the ability of lentiviral vectors to elicit an antibody-based protective immunity has, to date, not been evaluated. In the present study, we evaluated the potential of a lentiviral vector-based vaccine to elicit humoral immunity against West Nile virus (WNV). WNV is a mosquito-borne flavivirus that emerged in North America and causes encephalitis in humans, birds and horses. Neutralizing anti-WNV antibodies have been shown to be crucial for protection against WNV encephalitis. METHODS: The ability of lentiviral vector TRIP/sE(WNV), expressing the secreted soluble form of the envelope E-glycoprotein (sE(WNV)) from the highly virulent IS-98-ST1 strain of WNV, to induce a specific humoral response and protection against WNV infection was assessed in a mouse model of WNV encephalitis. RESULTS: Remarkably, a single immunization with a minute dose of TRIP/sE(WNV) was efficient at eliciting a long-lasting, protective and sterilizing humoral immunity, only 1 week after priming. CONCLUSIONS: This study broadens the applicability of lentiviral vectors as efficient non replicating vaccines against pathogens for which a neutralizing humoral response is one active arm of the protective immunity. The TRIP/sE(WNV) lentiviral vector appears to be a promising tool for veterinary vaccination against zoonotic WNV. PMID- 16308886 TI - Plasma fatty acids and lipoproteins in type 2 diabetic patients. AB - BACKGROUND: The dyslipidemia of type 2 diabetic patients is characterized by high VLDL, abnormal LDL composition and low HDL cholesterol concentrations. The aim of this study was to establish whether the type of dietary fats affects LDL size and density and HDL cholesterol concentrations in these patients. METHODS: Plasma phospholipid fatty acid composition, which reflects the type of dietary fatty acids, was quantified by gas chromatography. LDL relative flotation (LDL-Rf), a measure of LDL particle size and density, was determined by single vertical spin density gradient ultracentrifugation in 97 type 2 diabetic patients. RESULTS: By linear regression analysis of the data, plasma fatty acids were associated neither with LDL-cholesterol levels nor with LDL-Rf. The HDL cholesterol concentrations were negatively related with saturated fatty acids (r = -0.23; p = 0.02) but positively related with monounsaturated fatty acids (r = +0.20; p = 0.00). Furthermore, higher HDL concentrations were associated with large and buoyant LDL particles (HDL cholesterol vs LDL-Rf: r = +0.47; p = 0.00). In the multiple regression analysis, the LDL-Rf was significantly related both to triglycerides (beta coefficient = -0.55, p = 0.000) and HDL cholesterol (beta coefficient = 0.19, p = 0.034) concentrations. In a stepwise regression analysis including both triglycerides and HDL cholesterol, triglycerides alone explained the 43.0% of the LDL-Rf variability. CONCLUSIONS: A reduction of the dietary saturated fats and an increment of monounsaturated fats might increase HDL cholesterol concentrations in type 2 diabetic patients. Modifications of LDL composition might be expected from interventions aimed to reduce plasma triglycerides. PMID- 16308887 TI - Cancer-related communication between female patients and male partners scale: A pilot study. AB - BACKGROUND: The study's objective was to evaluate the reliability and validity of the Cancer-Related Communication Problems within Couples Scale (CRCP), a measure to assess whether patients and their partners have difficulty talking about cancer with each other. METHODS: The CRCP Scale included 18 items concerning open communication/emotional support, treatment-specific issues, self-protection, and protective buffering. Patients and partners responded through the American Cancer Society's website and mailed questionnaires through Y-ME. The CRCP's validity was tested using the Hospital Anxiety and Depression Scale (HADS) and the Dyadic Adjustment Scale (DAS, marital relationship). RESULTS: 189 female patients and 135 male partners participated in the survey. Three items were deleted based on lack of clarity or an item not being applicable to many respondents, resulting in a 15 item scale. The number of CRCP problems' internal consistency in the 15 item scale was good for both patients and partners (alpha coefficient = 0.87, 0.81, respectively). The number of patients' CRCP problems correlated with the DAS (p < 0.0001), and the number of partners' CRCP problems correlated with the HADS (p < 0.0001) and the DAS (p < 0.0001). CONCLUSION: Analyses supported the CRCP Scale's reliability and validity for female cancer patients and male partners. PMID- 16308888 TI - Application of the theory of planned behavior to understand intentions to engage in physical and psychosocial health behaviors after cancer diagnosis. AB - A cancer diagnosis can trigger change in both lifestyle behaviors and mental health outcomes such as 'growth' and 'benefit-finding'. Assuming changes in mental health outcomes are based upon changes in specific behaviors, the Theory of Planned Behavior (TPB) may facilitate understanding of post-diagnosis change in physical and psychosocial 'health' behaviors. Adults (n = 130) < or =2 years post-cancer diagnosis completed an internet survey. Current performance and future behavior intentions for two physical (e.g. eating a healthy diet) and four psychosocial (e.g. spending quality time with family/friends; engaging in spiritual or religious activities) health behaviors were assessed. TPB constructs (subjective norm, behavior attitudes, perceived behavioral control) for each of the six behaviors were also assessed. Multiple regression analyses indicated the set of TPB constructs accounted for an increment of 25-53% of variance in behavioral intentions beyond that accounted for by clinical and demographic variables. Among individual TPB constructs, behavioral attitude was most consistently associated with behavioral intentions while subjective norm was least consistently associated with behavioral intentions. The TPB could serve as a comprehensive model for understanding change in both physical and psychosocial health behaviors after cancer diagnosis and could suggest innovative approaches to developing interventions to enhance post-diagnosis 'growth' and 'benefit finding'. PMID- 16308889 TI - UN21(3-): a structurally characterized binary actinide heptaazide anion. PMID- 16308890 TI - Effects of stereo-regularity of multiblock co-poly(rac-lactide)s on stereo complex microparticles and their insulin delivery. AB - Uniform stereo-complex microparticles ranging from nanometer to micrometer size are prepared by using stereo multiblock co-poly(rac-lactide)s (smb-PLAs) with different stereo-regularity. At comparable molecular weights, as the smb-PLA stereo-regularity decreases from 88% to 76%, the crystallinity of the microparticles decreases noticeably, as proved by DSC and WAXD. At the same time, the shape of the microparticles varies from the flower shape to the sphere shape and the particle size increases markedly from 700-2700 nm as shown by SEM. However, all insulin-loaded microparticles are of cake-shape and their sizes depend on the stereo-regularity. The crystallization of smb-PLAs facilitated by insulin is evidenced by the increase of T(m) and DeltaH(f) in DSC. The highest insulin-loading content of 14.2% and -entrapment efficiency of 82.8% are obtained from the smb-PLA with the highest stereo-regularity of 88%. Release studies in vitro show the least first-day release at about 25% followed by continuous release of another 70% of insulin over one month. Stereo-complex microparticles of smb-PLAs with lower stereo-regularity resulted in a relatively lower insulin entrapment efficiency and -loading content, a larger first-day release, and also complete release of 90% of the total amount within one month. The release system follows a diffusion mechanism. By contrast, atactic PLA shows a very low entrapment efficiency of 16.7%. Structure of a stereo multiblock co-poly(rac lactide). PMID- 16308891 TI - Is second-trimester genetic amniocentesis for trisomy 18 ever indicated in the presence of a normal genetic sonogram? PMID- 16308892 TI - Uterine receptivity and the two and three dimensions of ultrasound. PMID- 16308893 TI - Evaluation and validation of a new risk score (CLEOPATRA score) to predict the probability of premature delivery for patients with threatened preterm labor. AB - OBJECTIVE: To develop a clinically useful tool to predict the probability of preterm delivery in patients with threatened preterm labor. METHODS: One hundred and seventy patients with preterm labor between 24 and 34 weeks of gestation were included. Preterm delivery < 37 weeks of gestation was the main endpoint of the study. The data were randomized and split into an evaluation set (n = 85) and a validation set (n = 85). The evaluation set was subjected to stepwise backward logistic regression analysis to quantify the relative impact of four potential risk factors, including individual patient factors, results of a rapid fetal fibronectin assay, and sonographic measurement of cervical length. Using the constant of the logistic regression analysis and the beta-coefficients for the identified risk factors the individual probability of preterm delivery for each woman of the validation dataset was calculated. The area under a receiver operating characteristics curve (AUC) was used to evaluate the discriminating power of the score. RESULTS: The overall rate of preterm delivery was 27.1%. The logistic regression analysis was performed for the potential predictors of spontaneous preterm delivery, identified by univariate analysis. These were positive fetal fibronectin, cervical length, previous preterm delivery and maternal age. Two risk factors were independent predictors of preterm delivery and were included in the CLEOPATRA I (clinical evaluation of preterm delivery and theoretical risk assessment) score: cervical length measurement and previous preterm delivery were associated with a higher risk of preterm delivery (odds ratio, 7.65 and 6.74, respectively). Since fetal fibronectin assay is not available at all institutions worldwide, it was excluded from the initial model. In the CLEOPATRA II model the risk factors fetal fibronectin and previous preterm delivery were associated with higher risk of preterm delivery, with odds ratios of 17.9 and 4.56, respectively. The discrimination power (AUC) obtained from the models were: CLEOPATRA I, 0.69 (95% CI, 0.56-0.82); CLEOPATRA II, 0.81 (95% CI, 0.69-0.93). CONCLUSION: In symptomatic women the risk for preterm delivery can be predicted best with the CLEOPATRA II score based on fetal fibronectin and previous preterm delivery. PMID- 16308894 TI - Cervical length at 18-22 weeks of gestation for prediction of spontaneous preterm delivery in Hong Kong Chinese women. AB - OBJECTIVE: To assess the value of a single cervical length measurement by transvaginal sonography (TVS) at the time of mid-trimester anomaly scan for predicting spontaneous preterm delivery (SPD) among Chinese women. METHODS: A prospective observational study was carried out involving 2880 subjects with singleton pregnancies and confirmed gestational age. Cervical length was measured at 18-22 weeks of gestation. RESULTS: The incidence of SPD < 34 weeks and < 37 weeks were 0.7% and 3.7%, respectively. Women with SPD < 34 weeks and SPD < 37 weeks had shorter median cervical lengths (32.6 mm and 36.2 mm, respectively) than those with term deliveries (37.6 mm) (P = 0.006 and 0.025, respectively). The predictive performance of cervical length was better for SPD < 34 weeks compared with < 37 weeks. A cervical length < or = 27 mm, which corresponded to the 4th centile, occurred in 36.8%, 62.5% and 100% of those with SPD < 34, < 30 and < 26 weeks, respectively. The positive likelihood ratio (LR) of a cervical length < or = 27 mm in predicting SPD < 34 weeks was 9.8. Using logistic regression, both short cervix and funneling were independent predictors for SPD < 34 weeks of gestation. The coexistence of funneling and a cervical length < or = 27 mm gave a positive predictive value (PPV) and LR of SPD < 34 weeks of 14.7% and 26.0, respectively. CONCLUSIONS: Mid-trimester cervical length is predictive of SPD in Chinese women. However, given the low PPV of a short cervical length, its clinical utility is still limited in low-risk populations. PMID- 16308895 TI - Ion motion in the rectangular wave quadrupole field and digital operation mode of a quadrupole ion trap mass spectrometer. AB - A quadrupolar electric field driven by a rectangular wave voltage can be used for mass-selective storage and analysis. The ion motion in such an electric field is derived, and the stability of ions is presented in the a-q diagram that is commonly used for sinusoidal wave quadrupole mass spectrometry in association with the solution of the Mathieu equation. The pseudo-potential well is discussed in an approximation that leads to the relation of secular frequency to operating parameters. A scheme for a digital ion trap mass spectrometer is described, based on this theory. An ion optics simulation was performed to check the theory of resonant ejection, and to prove the feasibility of the mass scan method for a practical ion trap of such geometry. PMID- 16308896 TI - Assessment of lung area in normal fetuses at 12-32 weeks. AB - OBJECTIVE: To establish reference intervals with gestation for the right and left lung areas and lung area to head circumference ratio (LHR). METHODS: This was a cross-sectional study of 650 normal singleton pregnancies at 12-32 weeks of gestation. We measured the left and right lung areas on the cross-sectional plane of the thorax, used for examination of the four-chamber view of the heart, by three different techniques: firstly, manual tracing of the limits of the lungs; secondly, multiplication of the longest diameter of the lung by its longest perpendicular diameter; thirdly, multiplication of the anteroposterior diameter of the lung at the mid-clavicular line by the perpendicular diameter at the midpoint of the anteroposterior diameter. RESULTS: The respective mean left and right lung areas (manual tracing) increased with gestational age, from 36 and 58 mm(2) at 12 weeks to 220 and 325 mm(2) at 20 weeks and 594 and 885 mm(2) at 32 weeks. This 16-fold increase in lung area was accompanied by a four-fold increase in head circumference. Consequently, the left and right LHR increased with gestational age. The most reproducible way of measuring the lung area was by manual tracing of the limits of the lungs and the least reproducible was by multiplying the longest diameter of the lungs by their longest perpendicular diameter. Furthermore, the method employing the longest diameter, compared with the tracing method, overestimated both the left and the right lung areas by about 45% and the method employing the anteroposterior diameter overestimated the area of the right lung by about 35%, but not that of the left lung. CONCLUSIONS: In the antenatal prediction of pulmonary hypoplasia by the assessment of lung area it is important to take gestational age into account. Dividing the lung area by the head circumference does not correct for the gestation-related increase in lung area. Reproducible measurement of the lung area is provided by manual tracing of the limits of the lungs, rather than by multiplication of lung diameters. PMID- 16308897 TI - Accuracy of fetal lung volume assessed by three-dimensional sonography. AB - OBJECTIVE: To determine the accuracy and precision of prenatal three-dimensional (3D) ultrasound in estimating fetal lung volume using the rotational multiplanar technique (VOCAL) by comparing it to postmortem volume measurements. METHODS: Fetal lung volume was measured during 3D ultrasound examination using a rotational multiplanar technique in eight cases of congenital diaphragmatic hernia (CDH) (six left and two right-sided) and in 25 controls without pulmonary malformation, immediately before termination. Prenatal 3D sonographic estimates of fetal lung volume were compared with postmortem measurement of fetal lung volume achieved by water displacement. RESULTS: The intraclass correlation coefficient of fetal lung volume estimated by 3D ultrasound and measured at postmortem examination was 0.95 in CDH cases and 0.99 in controls. Based on Bland Altman analysis, the bias, precision and limits of agreement were, respectively, 0.35 cm(3), 1.46 cm(3) and between -2.51 and + 3.21 cm(3) in cases with CDH and 0.08 cm(3), 2.80 cm(3) and between -5.41 and + 5.57 cm(3) in controls. The mean relative error of 3D ultrasound fetal lung volume measurement was -7.19% (from 42.70% to + 18.11%) in CDH cases and -0.72% (from -30.25% to + 19.22%) in controls, while the mean absolute error of 3D ultrasound fetal lung volume measurement was 1.40 (range, 0.71-2.52) cm(3) and 2.12 (range, 0.05-4.98) cm(3), respectively. Accuracy of 3D ultrasound for measuring fetal lung volumes was 84.86 (range, 57.30-99.48)% in cases with CDH and 91.38 (range, 69.75-99.45)% in controls. The mean intraobserver variability for lung volume estimated by 3D ultrasound was 0.28 cm(3) in controls and 0.17 cm(3) in CDH cases. CONCLUSION: Prenatal 3D ultrasound can estimate accurately fetal lung volume using the rotational multiplanar technique for volume measurements (VOCAL), even in fetuses with very small lungs, such as cases with isolated CDH. PMID- 16308898 TI - Fetal lung development in pregnancies of diabetic women assessed by quantitative ultrasonic tissue characterization. AB - OBJECTIVES: This study was performed to compare quantitative ultrasonic tissue characterization of the fetal lung at different gestational ages in uncontrolled diabetic patients with that in normal uncomplicated pregnancies. METHODS: A total of 44 women at 24-37 weeks' gestation with the diagnosis of diabetes in pregnancy were enrolled. Data were compared to those of the control group, which consisted of 140 women with uncomplicated pregnancies of the same gestational age. Longitudinal and transverse sections of the fetal thorax and upper abdomen were examined. A region of interest of constant size was defined and the tissue specific gray scale was determined by using interactive software. RESULTS: Compared with normal pregnancies, fetal lungs of diabetic pregnancies have a higher echogenicity between 28 and 37 weeks of gestation. The lung mean gray values (MGV) only differed significantly between 30 and 31 weeks of gestation in the group with diabetes (P = 0.033) compared to the control group. The MGV of the liver in diabetic and normal pregnancies is similar during pregnancy, significant differences being found only at 30-31 weeks of gestation (P = 0.038). The lung-to liver ratio in the control group showed a significant increase from 24 to 31 weeks and a slight non-significant decrease after 31 weeks. The ratio in the group with diabetes increased slightly up to week 33 and decreased slightly afterwards. CONCLUSION: Fetal lung MGV in uncontrolled diabetic pregnancies compared to that in uncomplicated pregnancies differs significantly only between 30 and 31 weeks of gestation. PMID- 16308899 TI - Analysis of Z-score distribution for the quality control of fetal ultrasound measurements at 20-24 weeks. AB - OBJECTIVES: To develop and evaluate the feasibility of Z-score distribution-based quality assessment for fetal biometry at 20-24 gestational weeks. METHODS: Four sonographers measured fetal biparietal diameter, head circumference, abdominal circumference and femur length in unselected pregnancies undergoing routine sonography at 20-24 weeks in our center between June 2001 and January 2004. All measurements were transformed into Z-scores. Distributions of Z-scores were compared with the expected standard normal distribution based on mean, SD and the Kolmogorov-Smirnov test. Modelling was performed to assess the sensitivity, specificity and Youden's index (sensitivity + specificity - 1) of each sonographer, reflecting their ability to detect fetuses with abnormal biometry in our population. RESULTS: A total of 5241 ultrasound examinations were included. None of the Z-score distributions could be considered similar to the standard normal distribution: means of Z-scores ranged from -0.604 to 0.73 and the SD ranged from 0.726 to 0.896. Sensitivity and specificity ranged from 25.31% to 68.67% and 93.44% to 100%, respectively; Youden's index ranged from 0.25 to 0.69. CONCLUSION: Analysis of Z-score distribution can be used to assess the quality of fetal measurements. Several indices can be extracted from this analysis in order to improve quality assessment of fetal biometry measured by ultrasound. PMID- 16308900 TI - Three-dimensional sonographic description of abnormal metopic suture in second- and third-trimester fetuses. AB - OBJECTIVE: To describe patterns of abnormal development of the metopic suture in association with fetal malformations during the second and third trimesters of pregnancy. METHODS: This was a cross-sectional study of the frontal bones and metopic suture in 11 fetuses at 17-32 weeks of gestation. Cases were selected because there were obvious abnormalities in the metopic sutures. In each case, a malformation was detected by two-dimensional (2D) ultrasound and the abnormality of the metopic suture was detected and evaluated on three-dimensional (3D) ultrasound, using transparent maximum mode. RESULTS: There were essentially four patterns of abnormality in the metopic suture: firstly, delayed development with a V- or Y-shaped open suture, which is found in normal fetuses at 12-16 weeks; secondly, a U-shaped open suture, presumably due to upward growth of the frontal bones with delayed closure; thirdly, premature closure of the suture, which is normally observed after 32 weeks; fourthly, the presence of additional bone between the frontal bones. Premature closure of the suture or additional bone between the frontal bones was observed in fetuses with holoprosencephaly and abnormalities of the corpus callosum, whereas the V-, Y- and U-shaped metopic sutures were observed in fetuses with facial defects involving the orbits, nasal bones, lip, palate and mandible, in the absence of holoprosencephaly and abnormal corpus callosum. CONCLUSIONS: This preliminary study describes the pattern of possible abnormalities of the metopic suture and should stimulate further investigation to establish the prevalence and evolution of abnormal sutures as well as the incidence and pattern of other associated defects. PMID- 16308901 TI - Diagnostic accuracy of varying discriminatory zones for the prediction of ectopic pregnancy in women with a pregnancy of unknown location. AB - OBJECTIVES: Various serum human chorionic gonadotropin (hCG) discriminatory zones are currently used for evaluating the likelihood of an ectopic pregnancy in women classified as having a pregnancy of unknown location (PUL) following a transvaginal ultrasound examination. We evaluated the diagnostic accuracy of discriminatory zones for serum hCG levels of > 1000 IU/L, 1500 IU/L and 2000 IU/L for the detection of ectopic pregnancy in such women. METHODS: This was a prospective observational study of women who were assessed in a specialized transvaginal scanning unit. All women with a PUL had serum hCG measured at presentation. Expectant management of PULs was adopted. These women were followed up with transvaginal ultrasound, monitoring of serum hormone levels and laparoscopy until a final diagnosis was established: a failing PUL, an intrauterine pregnancy (IUP), an ectopic pregnancy or a persisting PUL. The persisting PULs probably represented ectopic pregnancies which had been missed on ultrasound and these were incorporated into the ectopic pregnancy group. Three different discriminatory zones (1000 IU/L, 1500 IU/L and 2000 IU/L) were evaluated for predicting ectopic pregnancy in this PUL population. RESULTS: A total of 5544 consecutive women presented to the early pregnancy unit between 25 June 2001 and 14 April 2003. Of these, 569 (10.3%) women were classified as having a PUL, 42 of which were lost to follow up. Of the 527 (9.5%) cases with PUL analyzed, there were 300 (56.9%) failing PULs, 181 (34.3%) IUPs and 46 (8.7%) ectopic pregnancies. Overall, 74.6% were symptomatic and 25.4% were asymptomatic (P = 8.825E-07). The sensitivity and specificity of an hCG level of > 1000 IU/L to detect ectopic pregnancy were 21.7% (10/46) and 87.3% (420/481), respectively; for an hCG level of > 1500 IU/L these values were 15.2% (7/46) and 93.4% (449/481), respectively, and for an hCG level of > 2000 IU/L they were 10.9% (5/46) and 95.2% (458/481), respectively. CONCLUSIONS: Varying the discriminatory zone does not significantly improve the detection of ectopic pregnancy in a PUL population. A single measurement of serum hCG is not only potentially falsely reassuring but also unhelpful in excluding the presence of an ectopic pregnancy. PMID- 16308902 TI - Assessing clinical performance of gynecology residents: sonographic evaluation of adnexal masses based on morphological scoring systems. AB - OBJECTIVE: To assess the performance of inexperienced gynecology residents in the evaluation of adnexal masses using sonographic scoring systems. METHODS: From April 2003 to October 2004, transvaginal sonography was performed preoperatively by junior gynecology residents on 1-month shifts on 123 patients suspected of having adnexal masses. A total of 137 adnexal masses were found and evaluated using two different morphological scoring systems (those of DePriest and Lerner). The diagnostic performance of the sonographic scoring systems was assessed using the McNemar test and receiver-operating characteristics (ROC) curve analysis was used for lesion characterization. The diagnostic accuracy when the results of both scoring systems satisfied malignancy cut-off values was also investigated. RESULTS: Histopathological analysis revealed that, of the 137 lesions, 109 were benign, 23 were malignant and five were borderline. Best clinical cut-off levels were > or = 5 on the DePriest and > or = 3 on the Lerner scores. Both systems achieved good performance for characterizing malignancy. No significant difference was found in terms of the accuracy of the two systems as determined by mean areas under the ROC curves (0.816 and 0.783, P = 0.562). The combined approach using both scoring systems resulted in higher specificity (77.1%, P < 0.05) and positive predictive value without a significant decrease in sensitivity (82.1%) compared with either system alone. Of the 43 histologically confirmed false-positive cases, mature cystic teratoma was most common, with 13/22 (59%) cases being misinterpreted as malignancies. CONCLUSION: Junior residents, inexperienced at sonography, performed fairly in terms of evaluating adnexal masses with the help of morphological scoring systems. PMID- 16308903 TI - Giant omphaloceles with a small abdominal defect: prenatal diagnosis and neonatal management. AB - A giant omphalocele is a liver-containing protrusion through an abdominal defect wider than 5 cm in diameter. The giant form with a small abdominal wall defect is a rare condition which, to our knowledge, has not been described previously. We describe three cases with the typical features of elongated vascular liver pedicle and angiomatosis of the hepatic portal system. The abnormal liver organogenesis, due to extra-abdominal development, represented a significant risk factor for hepatic thrombosis after visceral reduction and liver rotation. All the neonates underwent surgery on the first day of postnatal life. One died because of a postoperative liver infarction, and the survivors needed prolonged respiratory support. Prenatal sonographic features, timing, delivery, type of surgical repair, and postnatal outcome are reviewed. A prenatal sonographic diagnosis could be useful to evaluate the abdominal ring and serial ultrasound examinations are recommended to detect promptly ominous signs of hepatic and bowel damage. Color Doppler may be useful to assess the anatomy of the abdominal vessels and their relationships with the herniated organs, although it was not used in any of the cases reported here. This congenital malformation might be considered as a pathological entity separate from giant omphalocele with large abdominal defect, with a severe prognosis due possibly to its different embryological development. PMID- 16308904 TI - Large infratentorial subdural hemorrhage diagnosed by ultrasound and MRI in a second-trimester fetus. PMID- 16308905 TI - Increased nuchal translucency thickness: a marker for chromosomal and genetic disorders in both offspring and parents. PMID- 16308906 TI - Palliative or curative treatment intent affects communication in radiation therapy consultations. AB - OBJECTIVE: To assess whether communication in radiotherapy consultations is affected by palliative or curative treatment intent. SUBJECTS AND METHODS: The study involved 160 patients and 8 radiation oncologists. Eighty patients visited the radiation oncologist (RO) for palliative treatment and 80 for curative treatment. The consultation prior to radiation treatment was analyzed with the Roter Interaction Analysis System (RIAS). Within three days, patients completed a quality of life questionnaire (EORTC-QLQ-C30). RESULTS: In palliative radiotherapy (PRT) consultations, ROs asked more (closed-ended) biomedical and psychosocial questions and provided more information on prognosis than in consultations for curative radiotherapy (CRT). Patients in both groups provided a great deal of psychosocial information but asked few questions. The ROs expressed more concerns in the PRT consultations, while patients did not. PRT patients received fewer supportive remarks than CRT patients. In both groups, explicit decision-making received little attention. Proxies who accompanied the patients took a more active role in PRT than in CRT consultations. CONCLUSION: Communication in radiotherapy is affected by treatment intent with respect to the main contents of the consultation. PMID- 16308907 TI - Multiple isolated intracardiac echogenic foci. Are they significant? PMID- 16308908 TI - Failure to diagnose a fetal anomaly on a routine ultrasound scan at 20 weeks. PMID- 16308909 TI - A virtual reality rendition of a fetal meningomyelocele at 32 weeks of gestation. PMID- 16308910 TI - Does felt age reflect health-related quality of life in cancer patients? AB - Subjective age is related to health and to life satisfaction. The present study on post-surgical cancer patients addressed two questions: (1) Do people differ in their health-related quality of life (HRQOL) depending on their felt age? (2) Is change in felt age associated with change in HRQOL? Data were collected at one month (T1) and six months (T2) after surgery (N = 159; 58% men, aged 24-86 years). At both waves, less than 14% of the patients felt older than their chronological age. Patients who felt younger than their age and patients who felt as old as they were reported better HRQOL than those subjectively feeling older. Between T1 and T2, an increment of felt age was found in 29% and a decrement in 28% of cases. HRQOL improved also among those who subjectively aged and among those whose felt age did not change, but in fewer aspects and with substantially lower effect sizes than among those whose felt age decreased. HRQOL is a correlate of felt age. However, an increment in felt age does not imply a decrease of HRQOL. PMID- 16308911 TI - Elevated release of sCD40L from platelets of diabetic patients by thrombin, glucose and advanced glycation end products. AB - The pro-inflammatory CD40/CD40L dyad participates in atherogenesis. Plasma levels of the soluble ligand (sCD40L) predict cardiovascular events and are elevated in diabetic patients. This study compared CD40/CD40L surface expression on platelets and T lymphocytes of diabetic and control subjects, and tested whether glucose and advanced glycation end products (AGEs) stimulate sCD40L release. Constitutive and inducible surface expression of CD40/CD40L on platelets or T lymphocytes did not differ between diabetic patients (n = 9) and controls (n = 13). Platelets from diabetic patients contained higher intracellular CD40L than controls (p < 0.05) and thrombin stimulated greater platelet sCD40L release in diabetic patients (15.11 +/- 16.77 ng/ml) compared to controls (3.64 +/- 2.03 ng/ml; p < 0.05). Glucose and AGEs induced platelet sCD40L release and CD40L expression in mouse megakaryocytes. This study demonstrates elevated CD40L content and inducible release from platelets of diabetic patients, and identifies glucose and AGEs as potential triggers of expression and release accounting for the elevated sCD40L plasma levels in these patients. PMID- 16308913 TI - Evaluation of cisplatin treatment given concurrently with pulsed irradiation in cisplatin sensitive and resistant human ovarian carcinoma cell lines. AB - The aim was to investigate the effect of combined treatment of cisplatin with acute or pulsed radiation in human ovarian carcinoma cells sensitive and resistant to cisplatin. Human ovarian cancer cell parental line A2780s and a derivative cisplatin-resistant line A2780cp were given cisplatin treatment before a single acute dose irradiation or concurrently during a pulsed-dose irradiation sequence. Cells were irradiated in the confluent state, trypsinized and plated after treatment. When the combined treatment for cisplatin was given before acute irradiation, the results showed additive to superadditive effects for both cell lines. However, the superadditivity was only significant in the sensitive cell line. For the concomitant treatment of cisplatin during pulsed-dose irradiation the results were additive, except for the highest cisplatin dose in the A2780cp line where subadditivity was observed. The results indicate that the combined treatments could be clinically useful even though the results are mostly not superadditive. However, high-dose cisplatin (3 microg ml(-1)) caused a subadditive effect in the resistant cell lines for pulsed irradiation. Thus, high dose cisplatin to overcome resistance is not effective. Cisplatin with both acute and pulsed irradiation showed additive effects indicating no advantage of using cisplatin in pulsed irradiation where sublethal damage repair may be greater. PMID- 16308914 TI - Effect of EGFR-inhibition on the radiation response of oral mucosa: experimental studies in mouse tongue epithelium. AB - The aim was to quantify the effect of selective inhibition of the epidermal growth factor receptor (EGFR) on the radiation response of mouse oral mucosa to daily fractionated irradiation. Irradiation comprised graded single doses of 25 kV X-rays to the lower tongue surface or fractionated doses of 5 x 3 Gy week(-1) (200 kV X-rays) over 1 or 2 weeks, followed by graded local doses, to generate full dose-effect curves. For selective inhibition of EGFR, BIBX1382BS, a tyrosine kinase inhibitor, was administered orally at a dose of 50 mg kg(-1), for the entire overall treatment time. The ED50 (the dose expected to induce ulcer in 50% of the mice) for untreated mucosa was 11.9 +/- 1.2 Gy. Fractionated irradiation administered over 1 or 2 weeks yielded ED50 values for the concluding test irradiation of 6.7 +/- 2.1 and 6.5 +/- 1.9 Gy, respectively. Administration of BIBX1382BS resulted in a non-significant increase of the top-up ED50 to 8.3 +/- 1.6 Gy (p = 0.1197) after 1 week and to 7.6 +/- 1.6 Gy (p = 0.2263) after 2 weeks. EGFR inhibition does not alter the radiation response of oral mucosa to fractionated irradiation or interfere with mucosal repopulation processes. This indicates that the regulation of mucosal repopulation is largely independent of EGFR activation. PMID- 16308915 TI - p53-Dependent accelerated senescence induced by ionizing radiation in breast tumour cells. AB - Ionizing radiation has been reported to promote accelerated or premature senescence in both normal and tumour cell lines. The current studies were designed to characterize the accelerated senescence response to radiation in the breast tumour cell in terms of its dependence on functional p53 and its relationship to telomerase activity, telomere lengths, expression of human telomerase reverse transcriptase (hTERT, the catalytic subunit of telomerase) and human telomerase RNA (hTR, the RNA subunit of telomerase), as well as the induction of cytogenetic aberrations. Studies were performed in p53 wild-type MCF 7 cells, MCF-7/E6 cells with attenuated p53 function, MDA-MB231 cells with mutant p53 and MCF-7/hTERT cells with constitutive expression of hTERT. Telomerase activity was measured by the telomeric repeat amplification protocol (TRAP assay), telomere lengths by the terminal restriction fragment (TRF) assay, hTR and hTERT expression by reverse transcriptase-polymerase chain reaction (RT-PCR), senescence by beta-galactosidase staining, and apoptosis by TdT-mediated d-UTP-X nick-end labelling (TUNEL assay). Widespread and extensive expression of beta galactosidase, a marker of cellular senescence, was evident in MCF-7 breast tumour cells following exposure to 10 Gy of ionizing radiation. Radiation did not suppress expression of either hTERT or hTR, alter telomerase activity or induce telomere shortening. Senescence arrest was also observed in irradiated MCF 7/hTERT cells, which have elongated telomeres due to the ectopic expression of the catalytic component of telomerase. In contrast to MCF-7 cells, irradiated MDA MB231 breast tumour cells and MCF-7/E6 cells failed to senesce and instead demonstrated a delayed apoptotic cell death. Irradiation produced chromosome end associated abnormalities, including end-to-end fusions (an indicator of telomere dysfunction) in MCF-7 cells, MCF-7/hTERT cells, as well as in MCF-7/E6 cells. When cells were maintained in culture following irradiation, proliferative recovery was evident exclusively after senescence while the cell lines which responded to radiation by apoptosis continued to decline in cell number. Accelerated senescence in response to ionizing radiation is p53 dependent and associated with telomer dysfunction but is unrelated to changes in telomerase activity or telomere lengths, expression of hTERT and hTR. In the absence of functional p53, cells are unable to arrest for an extended period, resulting in apoptotic cell death while accelerated senescence in cells expressing p53 is succeeded by proliferative recovery. PMID- 16308916 TI - Kraepelinian dichotomy. PMID- 16308917 TI - ECT in depression. PMID- 16308918 TI - Bibliography. Current world literature. Infectious diseases. PMID- 16308919 TI - Obesity and schizophrenia. PMID- 16308920 TI - Breast-feeding, bottle-feeding and Dr. Spock: the shifting context of choice. AB - In today's environment, breast-feeding represents both a medical gold standard for infant feeding and a moral gold standard for mothering. The morally charged character of this discourse makes the notion of choice in infant feeding particularly problematic and fraught with difficulty. From an historical content analysis of selected editions from 1946 to 1998 of Dr. Spock's famous child-care manual, this paper explicates the process through which the breast versus bottle discourse has shifted over the last half-century, and how these shifts have shaped the context of choice within which mothers must make their infant-feeding decisions. PMID- 16308921 TI - "Canada's Most Notorious Bad Mother": the newspaper coverage of the Jordan Heikamp Inquest. AB - This article is an examination of the media coverage of the inquest into the 1997 starvation death of an infant in a homeless women's shelter in Toronto. It explores the usefulness of using the discourse of mothering, intersected with the individualization of responsibility, to understand why one interpretation of this case seized the imagination of the public and informed media accounts. Examining how the mother of this infant was constructed as a "bad" mother helps to understand how hegemonic notions of "good" mothering are reinforced and perpetuated. The invocation of the ideals of motherhood, however, prevents us from seeing the reality of others' experiences of motherhood and the fact that, for many, there are barriers and restrictions to the ideal of good mothering. PMID- 16308922 TI - The ALHHS 30th anniversary celebration. PMID- 16308923 TI - 'Through thousands of errors we reach the truth' - but how? On the epistemic roles of error in scientific practice. AB - This essay is concerned with the epistemic roles of error in scientific practice. Usually, error is regarded as something negative, as an impediment or obstacle for the advancement of science. However, we also frequently say that we are learning from error. This common expression suggests that the role of error is not - at least not always - negative but that errors can make a fruitful contribution to the scientific enterprise. My paper explores the latter possibility. Can errors play an epistemically productive role in scientific research? The paper begins with a review of several twentieth-century approaches to error and the various agendas behind them. It is shown that only very few scholars have considered whether errors can be productive. The main part of the paper examines a concrete debate in early nineteenth-century on this material, the article offers some terminological clarifications of the common notion 'learning from error'. The conclusion argues that error can indeed play epistemically productive roles in scientific practice. PMID- 16308924 TI - Two chemists in two Koreas. AB - Taikyu Ree (1902-92) and Seung Ki Li (1905-96) were the most famous and influential scientists in twentieth-century Korea. Trained at Kyoto Imperial University during the Japanese occupation period (1910-45), both followed parallel career paths until 1945 but after that year those paths began to diverge. In 1948 Taikyu Ree moved to the University of Utah where he ran an informal graduate programme to train Korea chemists during the 1950s and 1960s. He became a model scholar in pursuit of knowledge for its own sake. In 1950 Seung Ki Li moved to North Korea where he succeeded in industrialising a new synthetic fibre from polyvinyl alcohol. He became a popular hero who demonstrated the utility of science. His success also helped Kim Il Sung, North Korea dictator, to consolidate his political ideology of Self-Reliance. Despite their different career patterns in very different circumstances, they shared something common; apolitical views and commitment to their work. An investigation of the careers of these two chemists illustrates the interwoven scientific, ideological and economic developments of South and North Korea during the second half of the twentieth century. PMID- 16308925 TI - [Early administration of vapreotide for variceal bleeding in patients with cirrhosis]. PMID- 16308926 TI - [From adenoma to carcinoma - how fixed are the paths of mutation?]. PMID- 16308927 TI - [Endoscopic reflux therapy by Endocinch]. PMID- 16308928 TI - [Is liver fibrosis a reversible event? Effect of pegylated interferon and ribavirin in patients with chronic hepatitis C]. PMID- 16308929 TI - [From fecal occult blood testing to Genoccult?]. PMID- 16308931 TI - [Change in paradigmas? From functional disorder to inflammation - associated disorder of the funktion activation of the mucosal immune system in irritable bowel syndrome]. PMID- 16308932 TI - [Biodegradable polyhydroxyalkanoates as carriers for antitumor agents]. AB - The possible use of biodegradable polyethers of microbial origin (polyhydroxyalkanoates) as matrices for deposition of daunorubicin (rubomycin), an antitumor anthracycline, was studied. The tablet dosage form of various rubomycin load (from 1 to 60% w/w) was prepared by cold compaction under pressure. The in vitro kinetics of the rubomycin release from the polymer matrix was investigated. It was shown that the rubomycin release to the medium resulted from the drug solution and diffusion within various periods, from tens hours to several weeks and months depending on the load. When the rubomycin load was under 20% w/w the drug release was prolonged and directly proportional to the observation time. When the rubomycin concentration was under 5%, the drug release kinetics corresponded to the type of the zero order reaction with prolonged release without sharp efflux at the initial stage of the observation. The findings showed that the polyhydroxyalkanoates were applicable as matrices for deposition of rubomycin and preparation of drugs with prolonged action. PMID- 16308933 TI - [Similarity and difference of the response of Peyer's patches and mesenteric lymph nodes of rats to polychemotherapy]. AB - The structure of the Peyer's patches and mesenteric lymph nodes of rats at different periods after polychemotherapy was investigated by light microscopy. After the use of antitumor drugs the number of the blasts and mytoses in the lymphoid follicles with the mesenteric lymph node bright centres and in the Peyer's patch follicles lowered, that along with the decrease of the size of the mantle zone in the lymph node follicles and the decrease of the area of the bright centres in the follicles of the lympoid formations in the intestinal wall was evident of the proliferation inhibition and B cells differentiation in the lymphoid organs. After the polychemotherapy the size of the germinative centres of the Payer's patch follicles decreased while the size of the mantle zone remained unchanged. The size of the mantle zone in the follicles of the mesenteric lymph nodes decreased while the size of the germinative centres did not change. The different responses of the lymphoid organs could be associated with some remote location of the lymph nodes with respect to the antigen source (damaged epithelium and intestinal lumen contents). PMID- 16308934 TI - [Experimental study on impact of salmon milt DNA on T-cellular immunity]. AB - The impact of a biologically active food supplement containing salmon milt DNA on cellular immunity was experimentally studied. It was shown that the DNA had a dose-dependent stimulating effect on lymphocyte transformation, stimulated the DTH response and protected from experimental listeriosis infection. PMID- 16308935 TI - [Chemotherapeutic effect of 3-(5-nitrofuryl)-7-(5-nitrofurfurylidene) 3,3a,4,5,6,7-hexahydro-2H-indazole in experimental staphylococcal infection]. AB - Chemotherapeutic activity of 3-(5-nitrofuryl)-7-(5-nitrofurfuryliden)-3, 3a, 4, 5, 6, 7-hexahydro-2H-indazol (compound 26) was studied on albino mice with experimental staphylococcal infection. The animals were contaminated intraperitoneally. The results of culture of the specimens of the organs of the mice killed within various terms and the values of the spleen bacterial index served as the criteria of the effectiveness. Compound 26 was administered in doses of 20, 10 and 5 mg/kg once a day for 3 days. The treatment was started simultaneously with or 24 hours after the contamination. The results showed that compound 26 in a dose 20 mg/kg (0.4 mg/mouse) possessed high chemotherapeutic activity in experimental staphylococcal infection of albino mice and could be recommended for a thorough study as a potential agent for chemotherapy of staphylococcal infection. PMID- 16308936 TI - [Influence of ABC-transporters on intracellular distribution of anticancer drugs and contribution of this index to multidrug resistance phenotype]. AB - A review of the literature on intracellular accumulation and distribution of anticancer drugs in sensitive and resistant tumor cells with the classic mechanism of multidrug resistance development, as well as on the role of ABC transporters in the processes is presented. According to the data discussed the authors prove that clinical analysis of human tumor multidrug resistance phenotype always needs separate estimation of the functional activity of ABC transporters, which control accumulation of MDR drugs in the cytoplasm and nucleus. The authors suggest a new term--"severity" of tumor phenotype of multidrug resistance. PMID- 16308937 TI - [Significance of determination of Shigella antibiotic resistance in bacteriological diagnosis of dysentery]. AB - The results of the Shigella antibiotic susceptibility assay within 1995-2002 are presented. 1472 cultures from 1158 patients with intestinal infections and bacteria carriers were isolated. The isolates were tested for their susceptibility to tetracycline, chloramphenicol, gentamicin, kanamycin, ampicillin and ofloxacin. It was shown that S. flexneri and S. sonnei were resistant to tetracycline. The S. flexneri isolates were highly resistant to chloramphenicol (73.3 to 96.0%) while resistance to it in the isolates of S. sonnei varied from 7.7 to 88.5%. In this connection the Levin medium with tetracycline was used to increase the Shigella isolation. In the study of the culture media efficiency with respect to isolation of Shigella it was observed that the Levin medium with tetracycline provided higher rates of S. flexneri and S. sonnei isolation (2.3- and 1.7-fold increase respectively) vs. the Shigella isolation on the Ploskirev medium without the antibiotic. PMID- 16308938 TI - [Etiology of severe hospital infections in intensive care units and antibiotic resistance of pathogens]. AB - The leading pathogens of severe infections in intensive care units were the following: respiratory tract infections--bacteria of the famility of Enterobacteriaceae (33.8%), Pseudomonas spp. (24.9%), Acinetobacter spp. (18.1%), Staphylococcus aureus (16.0%), blood flow infections--coagulase negative staphylococci (33.6%), S. aureus (26.1%), Enterobacteriaceae (17.6%), wound infections--Enterobacteriaceae (35.7%), coagulase negative staphyloccocci (17.8%), Pseudomonas spp. (14.3%). As for various species of Enterobacteriaceae, susceptibility was preserved in 91-100% of the isolates to meropenem, in 72-100% to cefoperazone/sulbactam, in 51-65% to cefepime, in 72-86% to amikacin, and in less than 50% to cephalosporins and fluoroquinolones. As for P.aeruginosa, 28% of the isolates was resistant to all the antibacterials, except polymyxin. The highest susceptibility to cefoperazone/sulbactam and meropenem was revealed in the isolates of Acinetobacter baumannii. Oxacillin resistance was detected in 64.9% of the S.aureus isolates. The oxacillin resistance as a rule was associated with resistance to macrolides, aminoglycosides and fluoroquinolones. As for coagulase negative staphylococci, oxacillin resistance was stated in 75.6% of the isolates. All the isolates of the Staphylococcus spp. preserved their susceptibility to vancomycin and linezolid. PMID- 16308939 TI - [Microbiological monitoring in prophylaxis and treatment of pyoinflammatory complications during early postoperative period in patients with cicatricial tracheostenosis]. AB - The species composition and antibiotic susceptibility of microflora isolated from patients with cicatricial tracheostenosis (CTS) and infectious processes were investigated. Schemes for the treatment of CTS with an account of the pathogen species and multidrug resistant staphylococcal infection are described. Comparative efficacy of the combined schemes (antibiotic + bacteriophage) for the therapy of pyoinflammatory processes in the patients with CTS and the efficacy of the treatment of the patients with CTS according to the traditional antibacterial schemes were studied. PMID- 16308940 TI - [Clinical efficacy of a novel antimicrobial drug combination containing ciprofloxacin and tinidazole in the treatment of patients with skin and soft tissue infections]. AB - Clinical and microbiological efficacies of a combined drug, a fixed combination of ciprofloxacin and tinidazole in the form of tablets (Cifran ST, Ranbaxy, India) were studied. The drug was given to 40 patients with skin and soft tissue infections in the complex surgical treatment. The clinical effect and bacteriological efficacy were observed in 97.5 and 75% of the cases respectively. The drug tolerability was good and no adverse reactions were stated. PMID- 16308941 TI - [Perspective technologies for drug design]. AB - The review present the literature data and the authors findings on perspective technologies for design of active chemical and natural compounds based on small molecules for highly specific therapy and correction of a wide variety of pathophysiological conditions in humans, that show overlapping development processes and have common mediators. The following strategies are discussed: chemogenomics strategy using computer screening libraries for generating of small molecule compounds with advantageous properties; strategy of chemokine network that provide control of many processes, from immunosurveillance to inflammation and from viral infections to cancer; strategy of using cytochrome P450 enzymes that guarantee maximum bioavailability of drugs and prevent their interaction and toxic effect; strategy of using superoxide dismutase enzymes (SOD) correcting superoxide anions overproduction in tissue injury and inflammation, from ischemia, organ transplantation to AIDS and cancer; strategy of telomer maintenance directed to anticancer and antiviral therapy as well as to correction of the aging processes; and strategy of short interfering RNAs capable to induce intracellular immunity to antigens of various origin. PMID- 16308942 TI - [Moxifloxacin, a novel extended spectrum fluoroquinolone in the treatment of severe infectious inflammatory diseases]. PMID- 16308943 TI - [Cephalosporins and community-acquired pneumonia: a choice alternative]. PMID- 16308944 TI - Gulf War illness. PMID- 16308945 TI - The expert witness and the lawyers. PMID- 16308946 TI - The molecular basis of prostate cancer. AB - Cancer involves accumulation of genetic alterations. This review highlights the alterations in control pathways for cell division, development, DNA repair, angiogenesis and cell death that are believed to be key players in the development of prostate cancer. PMID- 16308947 TI - Managing metastatic prostate cancer. AB - Prostate cancer is one of the most commonly diagnosed malignancies in the west. Most patients with metastatic or recurrent prostate cancer initially respond to androgen deprivation therapy, but almost all eventually progress. This review will focus on current treatment options for metastatic prostate cancer, with a focus on hormonal therapies, chemotherapy and treatment of bony disease, along with biological and targeted therapy. PMID- 16308948 TI - Prostate cancer: screening approaches. AB - Introducing screening for prostate cancer requires evidence that this would do more good than harm. Current evidence about the impact and natural history of prostate cancer, screening and diagnostic tests, and the effectiveness of treatments is reviewed below. PMID- 16308949 TI - Suicide prevention and audit. AB - Suicide is a devastating event and a major public health issue. On average one person dies every 2 hours in England as a result of suicide. It is the commonest cause of death in men under the age of 35 years. This article reviews the epidemiology, causal factors and prevention of suicide, and also briefly discusses the role of audit in this area. PMID- 16308950 TI - The electrocardiogram in pulmonary hypertension. PMID- 16308951 TI - Applying for a job. PMID- 16308952 TI - Interpretation of the abdominal radiograph: 1. PMID- 16308953 TI - Tracheostomy care. PMID- 16308954 TI - Injuries of the humeral shaft. PMID- 16308955 TI - Right iliac fossa pain. PMID- 16308956 TI - So you want to be ... an otorhinolaryngologist. PMID- 16308957 TI - Day case surgery: a modern view. AB - For several decades day surgery clinicians have championed the invaluable contribution that can be made to the provision of elective surgical care. It is now acknowledged as high impact change number one: treat day surgery rather than inpatient surgery as the norm for elective surgery. However, the potential is much greater and ambulatory practice will be the vanguard for modernizing the delivery of all surgical services of the future. It has been described as crucial to the future of health care. PMID- 16308958 TI - Urological emergency: diagnosis and management of renal trauma. AB - Renal trauma can be a life-threatening emergency and, because of its rarity, knowledge of its recognition and management is limited. This review discusses the clinical diagnosis, classification of injury, management and complications of this emergency condition. PMID- 16308959 TI - A pilot study of the feasibility of trainee hospital doctors undertaking significant event analysis. AB - Participating in significant event analysis is proposed as an important method of reflective learning that can enhance patient care and safety. Peer review is one way of making informed judgments on the ability of participants to apply the technique. PMID- 16308960 TI - Patent foramen ovale and paradoxical embolism presenting as acute coronary syndrome. PMID- 16308961 TI - Emery-Dreifuss muscular dystrophy with cardiac manifestations. PMID- 16308962 TI - Isotretinoin-related optic disc swelling. PMID- 16308964 TI - Metastatic prostate cancer presenting as diplopia with regression of signs with hormone manipulation. PMID- 16308965 TI - Competency training in anaesthesia. PMID- 16308967 TI - Phase in retirement with a flexible income. PMID- 16308966 TI - Implications and lessons learned from histopathology schools. PMID- 16308968 TI - The use of the cell saver in obstetrics. PMID- 16308969 TI - [A brief history of the International Agency for Research on Cancer (IARC)]. PMID- 16308970 TI - [Methods and techniques in of radical surgery for lung cancer. 1957]. PMID- 16308971 TI - [Heterotransplantation of malignant tumors using mammalian embryos. 1958]. PMID- 16308972 TI - [Stages of lymphoid tissue ontogenesis and histogenesis of non-Hodgkin lymphoma]. PMID- 16308973 TI - [Co-occurrence of hyperplasia, endometrial carcinoma and uterine myoma: role of sex hormones, their receptors and enzymes of estrogen metabolism]. PMID- 16308974 TI - [Prognostic and predictive factors in breast cancer]. AB - Recent data on prognostic and predictive significance of breast cancer-associated molecular markers are presented and their clinical value in making individual prognosis and choosing adequate therapy is discussed. Markers of tumor proliferation, steroid hormone receptors, factors of epidermal cell growth, plasminogen activators and inhibitors, factors influencing angiogenesis and apoptosis and genome characteristics are considered. It is concluded that only Ki 67, ER, PR and Her-2/neu may be recommended for clinical use. PMID- 16308975 TI - [Malignancies in the workers of the Koks Company (Kemerovo, Russia)]. PMID- 16308976 TI - [Natural killer cells (CD56, CD16) as a factor of local antitumor immunity in breast cancer]. AB - Two groups of malignancies with significantly distinct ratios of Ki-67 and CD16 expression were identified. Significant direct and inverse correlations between the markers were established (p < 0.01) whereas either group revealed two contradicting patterns of immune response. Whatever hormone receptor, natural killer cells (IKC) (CD16) levels correlated directly with tumor size and lymph node incidence (p < 0.01). IKC (CD56) did not contribute to breast tumor response. Our findings on CD16 expression may be used as criteria in prognosis prediction. PMID- 16308977 TI - [The microspectrophotometric criteria in estimating gastric mucosa dysplasia in non-invasive adenocarcinoma]. AB - Computer analysis was carried out of images of changes in area and ploidy of gastric mucous nuclei associated with concomitant pathology, dysplasia and non invasive adenocarcinoma of the stomach. In grave dysplasia, the area and ploidy of a nucleus were 213.7 +/- 3.42 microm2 and 10.2 +/- 0.2 c, and in well- or moderately differentiated cell adenocarcinoma--333.8 +/- 16.5 microm2 and 14.95 +/- 1.9 c, respectively. It is suggested that this evidence can contribute to a more accurate estimating of pathological processes taking place in the gastric mucosa when differential diagnosis between dysplasia and stomach cancer is undertaken. PMID- 16308978 TI - [The role of Ki-67, mutant suppressor gene p53 and low mitotic levels in defining the prognosis for adult granulosa cell tumor of the ovary]. AB - Clinico-morphological examination of adult granulosa cell tumors of the ovary was carried out in 17 patients (mean follow-up duration--146.2 +/- 22.16 months). Overall and relapse-free survival appeared significantly higher in low expression of Ki-67 (8.63 +/- 1.4%), mutant suppressor gene p53 (3.55 +/- 1.9%) and low mitotic levels in tumor (2.33 +/- 0.9 per 10 fields of microscope) (p < 0.05). PMID- 16308979 TI - [The RT-PCR-identified Fas and FasL expression and apoptosis in mouse hepatoma MH 22a and histiocytic sarcoma J-774 clonal lines coupled with in vitro cultivation with syngenic splenocytes]. AB - Malignant growth is associated with various patterns of interaction between tumor cells and those of the body immune system, interaction between Fas-receptor (Fas) and Fas-ligand (FasL) expression being one of them. These mechanisms were simulated in vitro using the main cell populations from murine hepatoma MH-22a, histiocytic sarcoma J-774 and their clonal lines obtained from cocultivation of tumor cells and syngenic splenocytes. Fas and FasL expression was identified by the RT-PCR method while apoptosis--by electrophoresis of low molecular DNA fractions and clonogenic survival. PMID- 16308980 TI - [The effect of vilon (Lys-Glu) on 1.2-dimethylhydrazine-induced neoplasia]. AB - The influence of vilon (Lys-Glu) on 1.2-dimethylhydrazine-induced neoplasia in mice has been studied. After treatment with 10 microg/kg, tumors developed in 14.3% which survived until first tumor detection (46 weeks) while, in control group, tumors occurred in 60%. Vilon-related inhibition of preneoplastic alterations in the kidneys was reported. PMID- 16308981 TI - [Endoscopic possibilities in the palliative treatment of cardial gastroesophageal cancer]. AB - Endoscopic surgery including chemical necrolysis, diathermy coagulation and excision, and argon-plasma was conducted in the course of two-stage palliative treatment of 31 patients with gastroesophageal cancer. A total of 273 procedures of tumor destruction were administered. Stenosis was either eliminated or significantly reduced in 28 (90.2%) on completion of stage I. Average stabilization duration was 44 days. During stage II, periodic destruction of tumor foci continued so that peroral feeding could be maintained for another 3 years. Mean survival was 8.7 months (one year--35%, two--20%, three--7.7%). Since no life-threatening complications were reported, our method may be recommended for out-patient hospital applications. PMID- 16308982 TI - [Clinical and radiological characteristics of telangiectatic osteosarcoma]. PMID- 16308983 TI - [Laparoscopic radical nephrectomy]. AB - Laparoscopic radical nephrectomy (left-sided--6, right-sided--3) was performed for T1b-3aN0-2M0-M1 renal tumor. Tumors measuring up to 12 cm were located in the mid or lower renal segment. Detailed descriptions of surgical procedure and chemical clearing to identify lymph nodes are presented. Surgery lasted 130-170 min. Intraoperative loss of blood was 50-150 ml. No postoperative complication was reported. Patients were discharged on days 5-7; time of disability ranged 21 30 days. PMID- 16308984 TI - [Ultrasound and hysteroscopic assessment as a component of diagnosis of genital tumors in women with menopausal bleeding]. PMID- 16308985 TI - [Arteropathy as late-onset complication of radiotherapy for malignant tumor]. AB - Such late-onset complications of radiotherapy as stenosis and occlusion of major arteries within fields of irradiation, designated in this paper as "post radiation arteropathy", have not received sufficient scrutiny. The study group of 50 patients (1964-2003) included 35 females and 15 males, aged 33-77, (mean age- 59 years). Total focal dose ranged 24-74.8 Gy (mean dose--48.9 Gy) per vascular bunch projection. Signs of ischemia were detected in the catchment area of irradiated artery after 8-360 months (average 90.6 months). Palliative treatment was given to 29 patients while 21 were operated on. Certain morphological features atypical of atherosclerosis were identified. Despite some negative factors, the results were not inferior to those of therapy for atherosclerosis. The hazard of stenotic effect of irradiation of major arteries in area of exposure should be perfectly clear to radiologists, oncologists and vascular surgeons. PMID- 16308986 TI - [Bronchial artery embolization for pulmonary bleeding in patients with lung cancer]. PMID- 16308987 TI - [Ultrasound pyrotherapy for tumors under the control of high-pole magnetic field]. PMID- 16308988 TI - The intensive statin therapy myth. AB - Statins play an important role in the treatment and prevention of coronary artery disease and atherosclerosis. Currently, however, despite its important qualities, the use of statin therapy in the treatment of CAD patients ranges only between 30 and 60% in Europe, the United States and Israel. A wide gap still exists between the numerous scientific publications demonstrating the beneficial effects of statins and the low rate of implementing the guidelines in practice. A Medline search up to June 2005 on all prospective, double-blind, randomized clinical trials evaluating the impact of intensive statin therapy (any statin dose >40 mg/daily) on clinical outcomes after a 1 year follow-up revealed only eight trials. In all the eight trials, with a follow-up period of 12-60 months, intensive statin therapy was significantly more effective than and at least as safe as placebo or other standard statin regimens. Thus, based on the current evidence-based medicine, intensive statin therapy enables more patients with CAD to achieve the current National Cholesterol Education Program goal for low density lipoprotein, while ensuring a relatively high safety profile. PMID- 16308989 TI - Effectiveness of establishing a dedicated acute stroke unit in routine clinical practice in Israel. AB - BACKGROUND: Clinical trials have demonstrated the superiority of managing acute stroke in a dedicated stroke unit over conventional treatment in general medical wards. Based on these findings, nationwide stroke unit care programs have been implemented in several countries. OBJECTIVE: To assess the effect of establishing a new dedicated acute stroke unit within a department of neurology on indicators of process of care and outcome of acute stroke in a routine clinical setting in Israel. METHODS: Stroke patients admitted to the Sheba Medical Center during the period March 2001 to June 2002 were included in a prospective study according to selection criteria. Data on demographics, risk factors, co-morbidities and stroke severity were collected. Indicators of process of care and outcome were assessed at hospital discharge and 30 days follow-up. Comparison between outcome variables by hospitalization ward was done using logistic regression analysis adjusting for confounders. RESULTS: Of 616 acute stroke patients (mean age 70 years, 61% men, 84% ischemic stroke), 353 (57%) were admitted to general wards and 263 (43%) to the stroke unit. Diagnostic procedures were performed more often and the infection rate was lower in the setting of the stroke unit. Poor outcome (modified Rankin scale > or = 3 or death) was present less often in patients managed in the stroke unit both at hospital discharge (adjusted odds ratio 0.5, 95% confidence interval 0.3-0.8) and at 30 day follow-up (adjusted OR 0.6, 95%CI 0.3-0.9). A Functional Independence Measure score < or = 90 or death at 30 day follow-up was less frequent among patients managed in the stroke unit than in general wards (adjusted OR 0.5, 95%CI 0.2-0.8). CONCLUSIONS: Improved outcomes and higher adherence to guidelines were observed in patients treated in a stroke unit within a department of neurology. The results suggest that patients with acute stroke should have access to treatment in a dedicated stroke unit. PMID- 16308990 TI - Who are the sedentary people in Israel? A public health indicator. AB - BACKGROUND: Increasing physical activity and thereby reducing a sedentary lifestyle can lower the risk of chronic diseases. Raising the population's involvement in physical activity is a major challenge for public health and healthcare services. OBJECTIVES: To identify subpopulations with a sedentary lifestyle and low levels of adherence to physical activity recommendations. METHODS: The Israel Center for Disease Control performed two national surveys during 2002-2003, interviewing 7,307 Jewish Israelis and 1,826 Arab Israelis over age 21. Respondents were asked if they engaged in physical activity lasting at least 20 consecutive minutes, and if so how frequently: less than once a week, once or twice a week, nearly every day or every day. RESULTS: Arab respondents were less physically active than Jewish respondents after adjusting for gender, age, level of religiosity, marital status, education, self-reported health, smoking, body mass index, and type of survey. Multiple logistic regression analysis run separately for Jews and Arabs found a more sedentary lifestyle, in both groups, among women, the less educated, those who were married and those with poor subjective health. Among Jews, younger age, increased religiosity, smoking and high BMI were associated with a sedentary lifestyle. CONCLUSIONS: The Jewish population is in need of more targeted and specific interventions for lower adhering subpopulations, such as women, the less educated and those with other risk factors. In the Arab population a more thorough understanding of the benefits of physical activity is needed; however, it seems that a general intervention is required to decrease the prevalence of a sedentary lifestyle all round. PMID- 16308991 TI - Effect of breast magnetic resonance imaging on the clinical management of breast cancer. AB - BACKGROUND: Mammography is the principal breast cancer imaging technique; however, sensitivity is reduced, especially in dense breast tissue. Magnetic resonance imaging is increasingly used in the detection and characterization of breast cancers. The high sensitivity (95-100%) of MRI is consistently observed, and in many situations, MRI is proving superior to classical forms of imaging. Assessment of its impact on management and outcome is vital if MRI is to become standard in the management of breast cancers. OBJECTIVES: To establish the impact of breast MRI on women undergoing testing in our institution. METHODS: We analyzed 82 cases that underwent MRI between January 2001 and April 2003. Analysis appraised the clinical impact of MRI testing in cases where medical summaries were available. RESULTS: Studies were categorized into five indications: a) screening in high risk women (n=7), b) search for primary disease in the presence of disease (n=5), c) monitoring of chemotherapy (n=2), d) postoperative assessment of tumor bed (n=9), and e) diagnostic/characterization of primary or recurrent breast cancer (n=59). Results were defined as negative, positive, or no impact on clinical management. MRI testing had a positive impact in 62 cases, affecting measurable change in 9 cases. Benefit was seen in screening, diagnosis and postoperative cases. In 15 cases, MRI stimulated investigations. CONCLUSION: MRI is a valuable tool in breast imaging and affects management. Further trials are necessary to clearly define the role of MRI and to ascertain whether in cases where beneficial impact on management is noted, there is ultimate impact on outcome. PMID- 16308992 TI - Antihypertensive effect of low calcium dialysis. AB - BACKGROUND: During maintenance hemodialysis acute elevation in serum calcium is common. Low calcium dialysis is advocated as a therapy for prevention of dialysis induced hypercalcemia. Approximately 16% of our chronic hemodialysis patients experience elevated arterial blood pressure during the hemodialysis session, becoming hypertensive by the end of the treatment. All these patients exhibited post-dialysis hypercalcemia. OBJECTIVES: To investigate the effect of low calcium dialysis on post-dialysis hypertension in view of an evident link between serum calcium and blood pressure in both normal renal function and chronic renal failure patients. METHODS: We evaluated 19 chronic hemodialysis patients in whom both post-dialysis hypertension and PDHCa were observed. We investigated changes in serum total calcium, ionized calcium, intact parathormone levels and arterial blood pressure in response to 4 weeks low calcium dialysis as a treatment for PDHCa. RESULTS: When PDHT patients were treated with low calcium dialysis, post dialysis blood pressure was significantly decreased compared to pre-dialysis values (155.3 +/- 9.7/82.2 +/- 7.9 mmHg pre-dialysis vs. 134.1 +/- 20.8/80 +/- 8.6 mmHg post-dialysis, P = 0.001). Additionally, post-dialysis blood pressure was significantly lower than post-dialysis blood pressure prior to the low calcium dialysis treatment (176.1 +/- 15/86 +/- 10.8 mmHg post-standard dialysis, 134.1 +/- 20.8/80 +/- 8.6 mmHg after low calcium dialysis, P = 0.001). A decline in post-dialysis serum calcium (2.34 +/- 0.2 vs. 2.86 +/- 0.12 mmol/L, P= 0.04) and ionized calcium (1.17 +/- 0.12 vs. 1.3 +/- 0.06 mmol/L, P = 0.03) compared to pre-dialysis levels was also achieved by this treatment, with no significant changes in iPTH levels. CONCLUSIONS: These data suggest a role for low calcium dialysis in treating acute serum calcium elevation and post-dialysis hypertension in patients receiving maintenance hemodialysis. PMID- 16308993 TI - Marked secular increase in the incidence rates of osteoporotic hip fractures in women and men in southern Israel. AB - BACKGROUND: For the last 35 years, our medical center has been the only referral center and provider of emergency medical services for a well-defined geographic area in southern Israel. OBJECTIVES: To evaluate trends in the incidence of hip fractures in this population. METHODS: The study was based on two surveys done approximately 20 years apart. It included women and men 50 years and older with radiographic evidence of a new hip fracture caused by low impact trauma. Only fractures that resulted from low or moderate trauma were considered for the current study. Incidence rates were calculated based on population data obtained from the official Central Bureau of Statistics. RESULTS: There was an overall twofold increase in the incidence rate of hip fractures. However, this increase occurred almost exclusively in the over-75 year old age groups (2.5-fold increase, both in women and men). The mean (and median) age of patients with hip fractures increased significantly over the study period, corresponding to the increase in longevity between the two periods. CONCLUSIONS: There was a marked secular increase in the incidence of proximal hip fractures in both genders, primarily because of an increase in the fracture rate in the very old. The increase in median age of fracture patients suggests that the observed increase in fracture rate can be attributed mainly to aging of the population rather than to deterioration in bone quality over the generations. PMID- 16308994 TI - Experience with somatostatin receptor scintigraphy in the management of pulmonary carcinoid tumors. AB - BACKGROUND: Somatostatin receptor scintigraphy has been used widely for the evaluation of neuroendocrine tumors in the gastrointestinal tract. Its use for detecting and staging thoracic carcinoids is only sporadically reported. OBJECTIVES: To evaluate the possible roles of SRS in the management of proven or suspected pulmonary carcinoids. METHODS: We conducted a retrospective study of all patients undergoing SRS for known or suspected pulmonary carcinoids in a tertiary referral center during a 10 year period. During this period 89 patients underwent resection of pulmonary carcinoids and SRS was used for detection, staging or localization purposes in 8 of them (9%). Scans were labeled true positive, true negative, false positive, or false negative in comparison with histologic or follow-up results. RESULTS: SRS was true positive in 6/6 lung locations; true positive in 2/8, true negative in 4/8 and false positive in 2/8 lymph node locations; and true positive in 1/8, true negative in 6/8 and false negative in 1/8 distant locations. The sensitivity, specificity, positive and negative predictive values and accuracy were 90%, 83%, 83%, 91% and 87% respectively. The scans were strongly positive in the tumors and involved lymph nodes. SRS correctly localized an occult secreting pulmonary carcinoid. Granulomatous and reactive lymph nodes showed increased uptake. SRS was accurate in ruling out distant metastases. CONCLUSIONS: SRS is effective for visualizing and localizing pulmonary carcinoids. It assists in the staging of these tumors by detecting lymph node involvement and confirming or ruling out distant metastases. Inflammatory areas in the lung or lymph nodes may be falsely positive. PMID- 16308995 TI - The epidemiology of primary biliary cirrhosis in southern Israel. AB - BACKGROUND: The epidemiology of primary biliary cirrhosis has changed significantly over the last decade, with a trend towards increasing prevalence in many places around the world. OBJECTIVES: To determine the overall prevalence of PBC in southern Israel and the specific rates for different immigrant groups between January 1993 and October 2004. METHODS: Multiple case-finding methods were used to identify all cases of PBC in the study region. Age-adjusted prevalence rates were compared among the different immigrant groups. RESULTS: A total of 47 cases of PBC were identified with an overall prevalence of 55 cases per million. All patients were women, and all except for a Bedouin Arab were Jewish. Foreign-born patients comprised 70% of our PBC cohort even though they represent only 45.4% of the regional population. This predominance of immigrants did not change when the rates were adjusted for age (P < 0.001). The prevalence rates were 40, 177, and 58 cases per million for those born in Israel, North Africa or Asia, and Eastern Europe, respectively. The age-specific prevalence rate for women older than 40 years varied from 135 cases per million among those born in Israel to 450 among immigrants from Eastern Europe and the former USSR to 700 cases per million among immigrants from North Africa and Asia. CONCLUSIONS: The prevalence of PBC in southern Israel is similar to that reported from some European countries. The rate is much higher among Jews than Arabs and among immigrants to Israel compared to native Israelis. PMID- 16308996 TI - Self-medication with antibiotics by a population in northern Israel. AB - BACKGROUND: The current study is part of a larger study--Self-Medication with Antibiotics and Resistance Levels in Europe (SAR project)--coordinated by the University of Groningen in the Netherlands and run in 19 European countries and Israel. OBJECTIVES: To estimate self-medication with antibiotics by a population in northern Israel. METHODS: We sent by post a questionnaire on antibiotic usage to 2,615 adults, both Jewish and Arab, living in northern Israel. RESULTS: The overall response rate was low (17.9%), particularly among the Arab population (9.4% of respondents). Among the 467 respondents, 169 (36.2%) reported 215 antibiotic courses within the last year. Amoxicillin was the antibiotic most commonly used (32.7% of courses); 89.4% of antibiotics were obtained via a physician's prescription; 114 respondents (24.4%) stored leftover antibiotics at home, and 81 (18.7%) would consider self-medication with antibiotics without a medical consultation. CONCLUSIONS: Over-the-counter acquisition of antibiotics is rare in Israel. However, the storage of leftover antibiotics in the home constitutes an alternative potential source of self-medication that can have untoward consequences, not only for the individual patient but also for the general population since inappropriate antibiotic usage contributes to the increasing rates of antimicrobial resistance. PMID- 16308997 TI - Adjustment disorder: a review of diagnostic pitfalls. AB - Adjustment disorder is a common diagnosis in psychiatric settings and carries a significant rate of morbidity and mortality. However, both current and previous diagnostic criteria are vague and lead to many difficulties in terms of validity and reliability. This review is based on a thorough literature search and a systematic evaluation of the empiric and theoretic data. The various pitfalls inherent in the process of diagnosing this disorder are discussed in light of the diagnostic criteria for the disorder. PMID- 16308998 TI - Issues to consider when using the new diagnosis of Mild Cognitive Impairment. PMID- 16308999 TI - High dose statin therapy for all patients with coronary artery disease? PMID- 16309000 TI - Dedicated stroke units and outcome. PMID- 16309001 TI - Multiple autoimmune disease in a patient with hyperprolactinemia. PMID- 16309002 TI - Lung abscess: an unusual complication of gastric banding. PMID- 16309003 TI - Weird activity and the wandering spleen. PMID- 16309004 TI - Sonographic features of a tophaceous nodule. PMID- 16309005 TI - Electrocardiogram changes in hyperkalemia: there and back again. PMID- 16309006 TI - Medial displacement of the internal carotid arteries in velocardiofacial syndrome. PMID- 16309007 TI - Generations at work. PMID- 16309008 TI - Postpolio syndrome--care in the workplace. PMID- 16309009 TI - Influenza vaccination: incidence of symptoms and resulting absenteeism in hospital employees. AB - A convenience sample of hospital workers, those receiving influenza vaccine and those not receiving vaccine, were asked to complete questionnaires delineating the occurrence of symptoms (e.g., fever, headache, extreme tiredness, dry cough, sore throat, runny nose, stuffy nose, muscle aches) and absenteeism in the 7-day period post-vaccination if vaccinated. Those unvaccinated completed the questionnaire in a self-selected 7 consecutive day period during the study conducted from November 2004 to February 2005. Those receiving either Fluzone or FluMist reported significantly fewer symptoms and related absenteeism than the unvaccinated group (p < .05). Administration of influenza vaccine did not result in higher rates of post-vaccination symptoms as compared to an unvaccinated group. Further, vaccinated employees did not experience higher absenteeism rates as a result of receiving either influenza vaccine. However, for those reporting absenteeism as a result of symptoms, mean absenteeism days were highest in the FluMist group (4.5 days) compared to the unvaccinated group (2.1 days) and the Fluzone group (1.9 days). PMID- 16309010 TI - Pulmonary tuberculosis in two employees at a skilled nursing facility: screening and treatment protocols. AB - In 2004, two employees in a skilled nursing facility developed active pulmonary tuberculosis (TB). Both employees were purified protein derivative (PPD) test positive for more than 10 years before developing the disease. Four employees converted to positive PPD status during the exposure investigation--two at baseline and two at the 12-week follow up. No additional cases of active TB were found. Several changes and additions have been made to the TB screening protocol after these two index cases were discovered. PMID- 16309011 TI - Injuries and assaults in a long-term psychiatric care facility: an epidemiologic study. AB - The objectives of this study were to document the high rates of acute injuries and physical assaults among nurses and certified nursing assistants working in long-term psychiatric care facilities and to identify risk factors for assaults and injuries to inform prevention strategies. A mixed-design cohort study was conducted. Acute injury and physical assault data were obtained from administrative records. Using staff rosters and schedule records, incidence rates were calculated by job title, gender, shift, and floor. Rates were also reported by severity, body part, type, and nature. Targeted interviews with staff members provided measures of physical lifting and resident combativeness. Injury rates were calculated by degree of lifting and assault rates were calculated by degree of resident combativeness. Overall rates of injuries (55.6 per 100 person-years) and assaults (67.3 per 100 person-years) were substantially higher than expected. Predictably, injuries were associated with resident lifting and assaults were associated with contact with combative residents. A higher risk of assault was found among women and higher risks of injury and assault were observed among full time employees compared to per diem or pool agency workers. In addition, weekend shifts were found to have a higher rate of injuries and a lower rate of assaults than weekday shifts. In similar long-term care facilities with psychiatric populations, efforts should be made to reduce lifting and avoid circumstances that agitate residents. Work organization factors should be taken into consideration when developing interventions. PMID- 16309012 TI - The diabetes prevention program: a worksite experience. AB - The purpose of this study was to determine if the U.S. National Institutes of Health Diabetes Prevention Program (DPP) could be successfully implemented in a worksite setting. Thirty-seven adult employees of BD Medical Systems of Sandy, Utah were enrolled in a single-group time-series study using the DPP. Two-hour oral glucose tolerance tests (OGTT) and other outcomes were measured at baseline, 6 months, and 12 months. Weight, body mass index, waist circumference, 2-hour OGTT, very low density lipoproteins, triglycerides, and aerobic fitness were significantly improved at 6 and 12 months and showed overall significant improvement across time. Fasting blood insulin, total cholesterol, low density lipoproteins, and total cholesterol/high density lipoproteins ratio were significantly improved at 6 months, but not at 12 months. Eighteen of the program participants (51%) were no longer in the pre-diabetes and diabetes categories after 1 year. Existing worksite health promotion and occupational health professionals can successfully offer the DPP and help employees improve glucose tolerance. PMID- 16309013 TI - Is gum disease related to cardiovascular disease? PMID- 16309014 TI - [Colorectal carcinoma]. PMID- 16309015 TI - [Nutrition in Switzerland 2002--results of the Swiss Health Survey]. AB - The 2002 Swiss Health Survey is the third survey conducted by the Federal Office of Statistics in intervals of five years. Data are collected of a random sample (n = 19706) of persons aged 15 and over, living in Switzerland: 29% are overweight (BMI 25-29.9 kg/m2), and 8% are obese (BMI 30+ kg/m2). 31% do not care about their nutrition. Less than daily consumption is reported by 34% for fruit, by 18% for vegetable, by 64% for milk, by 39% for milk products. 20% eat every day meat or meat products, 38% do rarely or never consume fish. Inadequate nutritional habits are observed more often in men, young people, and in participants with a low level of education. According to the Swiss Health Surveys of 1992, 1997, and 2002 nutritional habits have hardly improved during the last ten years in Switzerland. PMID- 16309016 TI - [About fruits and vegetables: the paradise does not come back]. AB - An increased fruit and vegetable consumption might be associated with a protection for the development of chronic diseases. The postulated mechanisms of this protection are multiple and no single mechanism can be identified. It is important to remember that the protection is mediated by the ideal combination of nutrients and phytochemicals in fruits and vegetables and not by a single chemical component. Accordingly it is more wise to eat fruits and vegetables instead of isolated compounds in pharmacological dosage. PMID- 16309017 TI - [About the value of video capsule endoscopy]. PMID- 16309018 TI - [A 69-year-old man with deep fatigue, weight loss and eosinophilia]. PMID- 16309019 TI - [*What do you do at the moment? I suffer"]. PMID- 16309020 TI - [B vitamins are not suitable for secondary prevention of cardiovascular disease]. PMID- 16309021 TI - [Drug and digestive probes. Aspects of pharmaceutical care]. PMID- 16309022 TI - [When the cough will not stop! Diagnosis and treatment of chronic cough]. PMID- 16309023 TI - [Treatment of sore throat pain with ambroxol-containing lozenges. Results of a pharmacy supported patient questionnaire]. PMID- 16309024 TI - [Height vertigo. Causes and prevention]. PMID- 16309025 TI - [Pimecrolimus and tacrolimus for neurodermatitis]. PMID- 16309026 TI - [Increased risk of for cardiovascular events from high doses]. PMID- 16309027 TI - [Smoke-free work places can reduce the risk of death]. PMID- 16309028 TI - [Backache. Good body condition is protective]. PMID- 16309029 TI - [Initial treatment can lighten low-severity community-acquired pneumonia]. PMID- 16309030 TI - [Little behavior improvement with memantine]. PMID- 16309031 TI - [Is depot formulation a long-term optimal treatment option?]. PMID- 16309032 TI - [Rofecoxib increases myocardial infarction risk in seniors]. PMID- 16309033 TI - [Do cholesterol ester synthetase inhibitors have preventive value?]. PMID- 16309034 TI - [Frequency of cesarean sections in four hospitals of Mexico City: 20 years later]. AB - BACKGROUND: This article is an update of a similar study published in 1984 in which the frequency of cesarean sections practiced in four Mexico City hospitals from 1976 to 1983 was analyzed. OBJECTIVE: To assess the frequency of cesarean sections practiced in four Mexico City hospitals from 1995 to 2002. MATERIAL AND METHOD: Data from these same hospitals corresponding to the 1995-2002 period was again collected. RESULTS: The frequency of cesarean sections in those four health care centers exceeded by far the internationally accepted standards; this frequency shows an increase, and it now accounts for twice as many operations as the number of interventions originally recorded. In 1976, 10 out of every 68 pregnancies came to end vaginally. By 2002, the same ratio was 10 out of 27. CONCLUSIONS: It is urgent to reduce the frequency of such a practice, as well as to reorganize the obstetric services; furthermore, the corresponding modifications in the training of such medical specialists must be done. PMID- 16309035 TI - [Frequency of postoperative seromas in mastectomy wounds with the use of one or two drainages: a pilot study]. AB - OBJECTIVE: To compare the number of postoperative seromas after mastectomy with the use of one or two drainages. PATIENTS AND METHODS: Women with malignant tumors who underwent Madden modified radical mastectomy were studied. They were divided into two groups: group A two drainages (axillar and pectoral), group B only one (axillar). For both groups demographical issues, body mass index (BMI = kg/m2), seromas and drainage period were determined. RESULTS: Twenty seven patients were evaluated: group A composed of 12 patients and B composed of 15. Age (years): A 56 +/- 14 vs B 50 +/- 6.5 (p NS), BMI: A 33.2 +/- 3.4 vs B 32.6 +/ 4.5 (p NS); seromas: A 4 vs B 5 (p NS); drainage period: A 23 +/- 6 vs B 24 +/- 7 (p NS). CONCLUSIONS: There are no differences in postoperative seroma development in mastectomy wounds with the use of one or two drainages. PMID- 16309036 TI - [Comparison of efficacy of combination of ketokonazol 400 mg + clindamycin 100 mg with ketokonazol 800 mg + clindamycin 100 mg in the treatment of vaginitis due to Candida and bacterial vaginosis]. AB - OBJECTIVE: To evaluate the effectiveness of the combination of ketoconazol 400 mg + clindamycin 100 mg for 6 days compared to ketoconazol 800 mg + clindamycin 100 mg in Candida vaginitis and bacterial vaginosis. MATERIAL AND METHODS: Patients aged 18-60 years, with clinical diagnosis of Candida vaginitis and vaginosis confirmed by culture of genital secretions were included. Patients were assigned at random to one of two treatment groups: group 1 was given ketoconazol 400 mg + clindamycin 100 mg during six days (K/C6D); group 2 received ketoconazol 800 mg + clindamycin 100 mg for three days + placebo during three days (K/C3D). Patients were evaluated at days 7 and 11 after initiating treatment, at day 11 culture of vaginal secretion was repeated; along with treatment and follow up period patients were asked to report presence of adverse events. RESULTS: Eighty-two patients were included, 41 in K/C6D group and 40 in K/C3D group. C. albicans was found at baseline in 19 patients in K/C6D group and in 15 in K/C3D group; at day 11 was cultured in 2/19 patients (10.52%) and in 2/15 (13.33%) (p = 0.626) respectively; G. vaginalis was cultured at baseline in 25 patients of each group, at day 11 was cultured in 1/25 patients (4.0%) of K/C6D group and in 4/ 25 (16.0%) of K/C3D group (p = 0.174). Clinical cure was found in 36/41 cases (87.8%) of K/C6D group and in 34/40 cases (85.00%) of K/C3D (p = 0.965) group. Only five patients presented adverse events, of which three were related to the treatment. CONCLUSION: Treatment of vaginitis and bacterial vaginosis with anyone of both formulations has the same clinical and microbiological effectiveness, and is well tolerated by the patient. The fact that K/C3D formulation is as effective as K/C6D has the advantage of shortening the treatment time of vaginal infections, and allows a better patient compliance. PMID- 16309037 TI - [Gestational trophoblastic disease. Experience at National Institute of Cancerology]. AB - INTRODUCTION: Gestational trophoblastic disease represents a variety of conditions that include hydatiform mole and choriocarcinoma. The common manifestation is high levels of beta human chorionic gonadotropin. In Mexico the incidence of the disease is 2.5 per 1,000 pregnancies. PATIENTS AND METHODS: This is a retrospective and descriptive analysis of patients with partial, complete or persistent hydatiform mole or choriocarcinoma diagnosis made from January 1988 to December 2003. We studied demographic characteristics, risk groups, treatment and response. We used descriptive statistics, multivariate analysis and Kaplan-Meier method for the survival analysis. RESULTS: We found 71 cases, the mean age at diagnosis was of 26 years, and 60.6% had choriocarcinoma. Vaginal bleeding was the most common manifestation at diagnosis. Thirty patients had low risk disease and 25 of them received chemotherapy based in methotrexate and folinic acid, 88% had complete response. In 10% of the cases the use of salvage chemotherapy showed a complete response. Overall survival was 100% at five years. Forty-one cases belonging to intermediate and high risk group were treated with chemotherapy (etoposide and actinomycin D in 68.3%). Overall response was of 90.2%, with complete response in 58.5% and partial response in 33.3%. Overall survival was of 94% at five years. Two cases developed second malignancies secondary to etoposide. CONCLUSION: Our results are similar to those reported in the literature. Overall survival in the low risk group was 100% and in the intermediate and high risk group of 94%. Etoposide and actynomicine D as first line chemotherapy had comparable results to those reported with EMA-CO and MAC. PMID- 16309038 TI - [Insulin sensitizers in the treatment of polycystic ovary]. AB - Polycystic ovary is characterized by anovulation, hyperandrogenemia and insulin resistance. Hyperinsulinemia is known to be associated with an increase in cardiovascular risk and the development of diabetes mellitus. The finding that insulin resistance has important implications in the pathogenesis of polycystic ovarian disease has elicit the concept of a therapeutic approach of insulin sensitizing drugs. Last decade multiple clinical trials about these drugs and upon genesis of polycystic ovary were designed; hence there is now sufficient evidence in the literature to support its clinical use. The management of polycystic ovary includes short-term objectives, such as treatment of infertility and control of androgen excess, as well as long-term considerations, such as prevention of endometrial cancer and management of dysmetabolic syndrome with its associated risk for developing type 2 diabetes and cardiovascular disease. The present review justifies the rationale use of insulin-sensitizing drugs in order to treat both short-term and long-term issues regarding polycystic ovarian disease. PMID- 16309039 TI - [Non-puerperal uterine inversion. Report of a case]. AB - Non-puerperal uterine inversion is extremely rare. We report a case associated with leiomyoma. It is presented the case of a multiparous of 46 year-old for presenting severe vaginal bleeding, pelvic pain and strange body vaginal sensation. The diagnosis was uterine myoma in abortive phase. An abdominal hysterectomy was performed to find a complete uterine inversion. This report represents an anecdotal case of non-puerperal uterine inversion successfully treated surgically. PMID- 16309040 TI - [Umbilical artery thrombosis. A report of a case and review of the literature]. AB - The thrombosis of the umbilical vessels is associated to high prenatal mortality. The arterial occlusion can compromise the maternal-fetal circulation with the neonatal death. Only two cases are reported in the literature with antepartum diagnosis and neonatal survival. We present the case of a 24 years old primigravida patient at 35 weeks normal pregnancy who was admitted in the hospital for diminution of the fetal movement's perception. The diagnosis was made with Doppler ultrasound of umbilical vessels in biophysical fetal evaluation; we decided pregnancy interruption with cesarean surgery. Cesarean delivery produced a male with Apgar scores of 8-9 at 1 and 5 minutes respectively, who was discharged one week later without vital compromise. PMID- 16309041 TI - [Cytologic method in the diagnosis and treatment of menopause. 1950]. PMID- 16309042 TI - [Impact of oral contraceptives as risk factors for papillomavirus infection and cervical intraepithelial neoplasia]. PMID- 16309043 TI - [60th anniversary of the AMGO]. PMID- 16309044 TI - [Clinical observation on acupuncture and moxibustion for treatment of aorto arteritis of arterial type in the head and arm]. AB - OBJECTIVE: To search for an effective therapy for aorto-arteritis of arterial type in the head and arm. METHODS: Eighty cases were randomly divided into an acupuncture and moxibustion group and a medication group, 40 cases in each group. Renying (ST 9), Taiyuan (LU 9), Jiquan (HT 1) and other acupoints were selected in the acupuncture group, and the medication group were treated with intravenous dripping of Fufang Danshen Liquid. Their therapeutic effects were observed. RESULTS: The clinical cured rate and the total effective rate were 15.0% and 95.0% in the acupuncture group, and 0 and 75.0% in the medication group, respectively, with a very significant difference between the two groups in the therapeutic effect (P<0.01). CONCLUSION: Acupuncture and moxibustion has a definite therapeutic effect, which is significantly superior to that of the routine medication for aorto-arteritis of arterial type in the head and arm. PMID- 16309045 TI - [Observation on therapeutic effect of pulse magnetic acupuncture at scalp acupoints on acute cerebral infarction]. AB - OBJECTIVE: To apply pulse magnetic acupuncture at scalp acupoints to treat acute cerebral infarction and to explore the mechanism. METHODS: A pulse magnetic acupuncture group, a routine acupuncture group and a static magnetic acupuncture group were set up, 30 cases in each group. Their clinical therapeutic effects were observed. RESULTS: The cured-markedly effective rate was 80.0% in the pulse magnetic acupuncture group and 70.3% in the routine acupuncture group with no significant difference between the two groups (P>0.05), which were significant difference with 36.6% in the static magnetic acupuncture group (P<0.01). CONCLUSION: Pulse magnetic acupuncture and routine acupuncture at scalp acupoints have same therapeutic effect on acute cerebral infarction, which is superior to that of static magnetic acupuncture. PMID- 16309046 TI - [Observation on therapeutic effect of electroacupuncture on obesity induced by antipsychotics]. AB - OBJECTIVE: To find out an effective therapy for obesity induced by antipsychotic agents. METHODS: One hundred and one cases of obesity who were being treated with antipsychotic agents were randomly divided into an observation group and a control group. The observation group were treated with electroacupuncture at Quchi (LI 11), Zusanli (ST 36), Fenglong (ST 40), Shangjuxu (ST 37) and Xiajuxu (ST 39), once a day for 8 weeks; and no treatment was given to the control group. The Brief Psychiatric Rating Scale (BPRS) was used for assessment of therapeutic effect and the Treatment Emergent Symptom Scale (TESS) for adverse effects. RESULTS: The clinically effective rate of the observation group (54.9%) was superior to the control group (10.0%) with a significant difference between the two groups (P<0.05). The BPRS score-reducing rate was 24.92% and 28.62% in the both groups respectively, and with no adverse effects. CONCLUSION: Electroacupuncture has a good therapeutic effect on obesity induced by antipsychotic agents, and it improves the patient's compliance with no adverse effect. PMID- 16309047 TI - [Therapeutic effect of electroacupuncture combined with medicine on acute rheumatoid arthritis at active stage]. AB - OBJECTIVE: To explore a treatment program for increasing therapeutic effect on rheumatoid arthritis at active stage. METHODS: One hundred and forty-six cases were randomly divided into treatment group (n = 74) and medicine control group (n = 72). The treatment group were treated by electroacupuncture at Quchi (LI 11), Hegu (LI 4), Yanglingquan (GB 34), etc. , combined with meloxicam, sulfasalazine and MTX. The control group treated by simple the Western medicines. Their therapeutic effects were compared. RESULTS: The effective rate was 79.73% in the treatment group and 51.39% in the control group with a significant difference between the two groups (P< 0. 05). CONCLUSION: Electroacupuncture combined with medicine has a better therapeutic effect than the simple medicine on rheumatoid arthritis at active stage. PMID- 16309048 TI - [Evaluation of therapeutic effect of acupuncture at Jiaji (EX-B 2) on hyperosteogeny of lumbar vertebrae]. AB - OBJECTIVE: To assess therapeutic effect of acupuncture at Jiaji (EX-B 2) on hyperosteogeny of lumbar vertebrae. METHODS: Seventy-two cases were treated with Huatuo jiaji (EX-B 2) of L2 approximately L5 as main acupoints, and their clinical therapeutic effects were assessed with Oswestry Disability Index (ODI). RESULTS: ODI was 0.562 +/- 0.199 before treatment and 0.123 +/- 0.196 after treatment with a significant difference before and after treatment (P<0.01), and the effective rate was 94.4%. CONCLUSION: Acupuncture at Jiaji (EX-B 2) can improve functional activity for the patient of hyperosteogeny of lumbar vertebrae. PMID- 16309049 TI - [Observation on therapeutic effect of warming acupuncture and moxibustion combined with Chinese drugs on ovarian cysts]. AB - OBJECTIVE: To observe the therapeutic effect of acupuncture and moxibustion combined with Chinese herbs on ovarian cysts. METHODS: Forty-six cases were randomly divided into a treatment group of 26 cases and a control group of 20 cases. The treatment group were treated with warming acupuncture and moxibustion at Zigong (EX CA 1), Qihai (CV 6), Sanyinjiao (SP 6), etc. and oral administration of Chinese herbs Quyu Decoction, and the control group were treated with Quyu Decoction only. One month later, their therapeutic effects were observed. RESULTS: The cured rate and the effective rate were 53.8% and 88.4% in the treatment group, and 15.0% and 85.0% in the control group, respectively, the cured rate in the treatment group being significantly better than that in the control group (P<0.01). CONCLUSION: Warming acupuncture plus moxibustion combined with oral administration of Quyu Decoction can significantly increase the cured rate for ovarian cysts. PMID- 16309050 TI - [Comparison of therapeutic effects of acupuncture and moxibustion on diabetic peripheral neuropathies]. AB - OBJECTIVE: To observe and compare therapeutic effects of acupuncture, acupoint injection and tapping with plum-blossom needle and its influences on the nerve conduction velocity in the patient of diabetic peripheral neuropathies. METHODS: Fifty-four cases were randomly divided into 3 groups. The group I were treated by the basic treatment plus acupuncture, the group II by the basic treatment plus acupoint injection of methycobal, and the group III by the basic treatment plus tapping with plum-blossom needle. RESULTS: The effective rate was 85.71% in the group III, 76.47% in the group II and 56.25% in the group I; after treatment MCV and SCV improved significantly in the group III. CONCLUSION: The basic treatment plus tapping with plum-blossom needle has a better therapeutic effect on diabetic peripheral neuropathies. PMID- 16309051 TI - [Effects of acupuncture on diabetic peripheral neuropathies]. AB - OBJECTIVE: To observe therapeutic effect of acupuncture on diabetic peripheral neuropathy (DPN) and explore the mechanism. METHODS: Sixty-eight cases of DPN were treated with acupuncture at Jianyu (LI 15), Quchi (LI 11), Waiguan (TE 5) etc. and their blood sugar, blood lipids and hemorrheologic indexes and nerve conductive velocity before and after treatment were determined. RESULTS: After treatment, 43 cases were markedly effective, 20 cases were effective and 5 cases were ineffective, with a total effective rate of 92.6%; the nerve conductive velocity, plasma insulin, C-peptide, opioid peptides increased significantly (P<0.01) and blood viscosity significantly decreased (P<0.01). CONCLUSION: Acupuncture has a definite therapeutic effect on DPN and the increase of nerve conductive velocity is possibly related with the increase of plasma opioid peptides. PMID- 16309052 TI - [Clinical observation on acupuncture by stages combined with exercise therapy for treatment of Bell palsy at acute stage]. AB - OBJECTIVE: To find out a method for increasing clinical therapeutic effect on Bell palsy at acute stage. METHODS: Ninety cases of Bell palsy were randomly divided into an observation group, a control group I and a control group II, 30 cases in each group. They were treated respectively with acupuncture plus exercise therapy, simple acupuncture therapy, and simple exercise therapy, and their therapeutic effects were observed. RESULTS: The cured rate was 66.7% in the observation group, 53.3% in the control group I and 46.70% in the control group II, the observation group being better than the two control groups (P<0.05). CONCLUSION: Acupuncture by stage combined with exercise therapy can increase the therapeutic effect on Bell palsy at acute stage, and it is a better therapy for Bell palsy. PMID- 16309053 TI - [Therapeutic effect of deep acupuncture at local acupoints on trigeminal neuralgia]. AB - OBJECTIVE: To search for an effective method for increasing therapeutic effect on trigeminal neuralgia. METHODS: Ninety cases of primary trigeminal neuralgia were randomly divided into a deep needling group and a routine needling group, 45 cases in each group. The routine needling group were treated by shallow acupuncture at local acupoints and distal acupoints along the Hand and Foot Yangming Channels, and the deep needling group were treated by acupuncture at the above acupoints and deeply needling at the local acupoints to nerve stem for 3 courses. RESULTS: In the deep needling group 12 cases were clinically cured, 24 cases were markedly effective, 7 cases improved and 2 cases were ineffective, with a total effective rate of 95.6%; and in the routine needling group, the corresponding figures were 7, 15, 12, 11, 75.6%. The therapeutic effect in the deep needling group was better than that in the routine needling group (P<0.05). CONCLUSION: Deeply needling local acupoints plus acupuncture at distal acupoints along the Hand and Foot-Yangming Channels can increase significantly the therapeutic effect on trigeminal neuralgia. PMID- 16309054 TI - [Clinical observation on acupuncture combined with cupping therapy for treatment of ankylosing spondylitis]. AB - OBJECTIVE: To search for an effective method for treatment of ankylosing spondylitis (AS). METHODS: The treatment group (n = 32) were treated by acupuncture combined with cupping therapy and the control group (n = 30) by simple acupuncture for 40 days. RESULTS: The clinical remission rate was 62.5% in the treatment group and 33.3% in the control group with a significant difference between the two groups (P<0.01); the total effective rate in the treatment group was 93.8% which was better than 83.3% in the control group (P<0.01); the recurrence rate after one year was 3.3% in the treatment group and 24.0% in the control group with significant difference between the two groups (P<0.01). CONCLUSION: Acupuncture combined with cupping therapy in the therapeutic effect on ankylosing spondylitis is better than simple acupuncture, with shorter therapeutic course and lower recurrence rate. PMID- 16309055 TI - [Comparative study on therapeutic effects of Baihui (GV 20), Shuigou (GV 26) and Shenmen (HT 7) in treatment of vascular dementia]. AB - OBJECTIVE: To compare the relative characteristics of Baihui (GV 20), Shuigou (GV 26) and Shenmen (HT 7) in treatment of vascular dementia. METHODS: Fifty cases of vascular dementia (VD) were divided into 5 groups randomly. The patients in the group A were treated by acupuncture at routine acupoints. Besides the routine acupoints of the group A, Baihui (GV 20), Shuigou (GV 26) and Shenmen (HT 7) were added to treat the patients in group B, C, D, respectively, while Baihui, Shuigou and Shenmen were added to the group E. The clinical symptoms and signs were observed. RESULTS: Baihui and Shenmen can improve memory, orientation, reaction, obstinacy and trance of VD patients. Baihui is good at helping the patients to understand, calculate and adapt the society. Shuigou is good at improvement of sleeping, slow reaction, trance and memory of the patients. The combination of Baihui, Shuigou and Shenmen can generally promote intelligence level and society adaptive ability. CONCLUSION: Baihui, Shuigou and Shenmen have own characteristics in improving clinical signs and intelligence of patients of VD, and the combination of the 3 acupoints has the best effect in treating VD. PMID- 16309056 TI - [Effect of acupuncture on expression of connexion 43 in "Zusanli" (ST 36) of the rat]. AB - OBJECTIVE: To observe effects of acupuncture at "Zusanli" (ST 36) and non-channel non-acupoint on expression of connexion 43 in the rat, so as to probe into possible relationship between the acupoint and connexion 43 (Cx43). METHODS: Twenty healthy SD rats were randomly divided into a non-acupuncture group and an acupuncture group, 10 rats in each group. Quantity and distribution of Cx43 on skin tissue of "Zusanli" and non-channel-non-acupoint were detected by immunohistochemistry with a fluorescence double-label technique and laser scanning confocal microscope. RESULTS: Cx43 was mainly expressed in epidermis, hair follicles and other epithelial keratinocytes; the expression of Cx43 in the acupoint was significantly higher than that at the non-channel-non-acupoint in the non-acupuncture group (P<0.01); after acupuncture at acupoint expression of Cx43 in the acupuncture group significantly increased, with a very significant difference as compared with that in the acupoint in the non-acupuncture group (P<0.01). CONCLUSION: Connexion 43 and gap junctions in acupoint are possibly the main components of acupoints, and acupuncture can significantly increase expression of connexion 43 in acupoints. PMID- 16309057 TI - [Effect of electroacupuncture on proliferation of astrocytes after spinal cord injury]. AB - OBJECTIVE: To explore the mechanism of acupuncture in treatment of spinal cord injury (SCI). METHODS: Adult Wistar rats were used to make SCI model by Allen's method. The SCI rats were treated with electroacupuncture (EA) at acupoints of the Governor Vessel for 3 days, 1 week, 2 weeks or 4 weeks. Normal group and spinal cord injury group were used as controls. The number and morphology of astrocytes in each group were investigated by electron microscope, immunohistochemistry and in situs hybridization methods. The expression of glial fibroblast acid protein (GFAP) mRNA in the injured spinal cord was detected by reverse transcription polymerase (RP-PCR). RESULTS: The mitochondria and ribosomes of astrocytes in the EA group increased. The number of astrocytes increased after SCI, in gray matter being more than that in the white matter, in the caudal being more than that in the rostral. The expression of GFAP mRNA in the EA group was significantly lower than that in the control group. CONCLUSION: Electroacupuncture can inhibit the reactive proliferation of the astrocytes after spinal cord injury and prevent formation of the glial scar. PMID- 16309058 TI - [Effect of acupuncture on expression of neurology-related genes in the brain tissue of the rat]. AB - OBJECTIVE: To analyze and explore the molecular mechanism of acupuncture precondition (AP) in improving brain ischemia reperfusion injury. METHODS: Six male Wistar rats were divided into a normal group of 3 rats and an acupuncture group of 3 rats. In the acupuncture group, acupuncture precondition was first given at "Shenshu" (BL 23) and "Baihui" (GV 20). The brain of the rat was removed 0.5 hour after the needle was withdrawn, and then gene differential expression was analyzed by genechip method. RESULTS: After acupuncture precondition for 0.5 h, expression of 265 genes showed changes of more than 2 times, and that of 20 genes had a significant difference as compared with that of the normal rat (P<0.05). Among the genes, expression of 8 was upregulated and 12 downregulated; 11 were related with neuron signal conduction (ion channel, transmitter, receptor, second message), 5 with gene transcription and control, 2 with metabolism enzyme, 1 with heat shock reaction and 1 with cell skeleton. CONCLUSION: Some gene expression substances induced by acupuncture play an important role in AP resisting brain ischemia reperfusion injury; and functional changes of neurons after AP are possibly the important basis of its subsequent action in resisting brain ischemia reperfusion injury. PMID- 16309059 TI - [Introduction to clinical studies of acupuncture and moxibustion of Britain Foundation of TCM]. AB - In Western countries, more and more people select acupuncture and moxibustion as medical ways, and more and more acupuncture and moxibustion research organizations are set up. Britain TCM foundation is such one center for acupuncture research. This center is aimed at promoting acupuncture research. Since 1991, a series of clinical trials and several big research tasks have been completed. The clinical designs in the center are very careful and scientific, being worth to be learnt. In the paper, clinical studies in the center are introduced for increasing levels of acupuncture and moxibustion research at home. PMID- 16309060 TI - [From Xingnao Kaiqiao needling method to construction of Shi's apoplexy unit]. AB - According to recognition of TCM for pathogenesis of ischemic apoplexy, Shi's apoplexy unit therapy is founded on the basis of Xingnao Kaiqiao needling method and apoplexy unit. This method covers therapeutic methods of apoplexy at different stages and different periods, forming a complete comprehensive therapeutic program for apoplexy, including ICU treatment at the initial stage, operation treatment, acupuncture and moxibustion, TCM and WM treatment combined with rehabilitation training, diet therapy, mental and psychological therapies and healthy education, etc. The marked therapeutic effect of Shi's apoplexy unit is not due to a single therapy, but the comprehensive therapy. PMID- 16309061 TI - [Study of music electroacupuncture]. AB - OBJECTIVE: To explore characteristics and advantages of music electroacupuncture instrument. METHODS: Investigate clinical therapeutic effect and action mechanisms of the music electroacupuncture instrument, the relation of music therapy with improving looks and genes, and the links of music therapy with theories on 14 channels to prove its characteristics and advantages. CONCLUSION: Music electroacupuncture can overcome the decline of late therapeutic effects of pulse electroacupuncture and its disadvantages, beat of the local tissue of electroacupuncture. It is a generation of electroacupuncture of a new type. PMID- 16309062 TI - [Chronomedicine, 6-meridian syndrome differentiation theory and navel needling therapy]. AB - OBJECTIVE: To explore the relation among navel needling therapy, chronomedicine and 6-meridian syndrome differentiation theory. METHODS: Adopt TCM circular motion ascending and descending theories of functional activity of qi and observe clinical application of navel earthly branch needle inserting method. RESULTS: The navel earthly branch needle inserting method had a very good therapeutic effect on the diseases with obvious time attack regularity. CONCLUSION: Chronomedicine is applicable for traditional medicine and navel needling therapy. PMID- 16309063 TI - [Regulative action of acupuncture and moxibustion on mucosal immunity]. AB - OBJECTIVE: To explore the regulative action of acupuncture and moxibustion on mucosal immunity and its curative effect on mucosal relative diseases. METHODS: Review the recent 10 years' achievements of studies on mucosal immunity and analyze the regulative action of acupuncture and moxibustion on the mucosal immunological system in treatment of mucosal relative diseases. CONCLUSION: Acupuncture and moxibustion has a good regulative action on local mucosal immunological system, which is one of the mechanisms of acupuncture and moxibustion in prevention and treatment of mucosal relative diseases. This proves a more reliable basis for treatment of mucosal relative diseases with acupuncture and moxibustion. PMID- 16309064 TI - Never underestimate the value of teachers. PMID- 16309065 TI - Using systematic reviews for evidence-based practice. PMID- 16309066 TI - Surgical treatment of obstructive sleep apnea. AB - THE PRIMARY TREATMENT for obstructive sleep apnea (OSA) has been continuous positive airway pressure (CPAP) therapy, but the minimum acceptable number of hours of nightly CPAP use remains unclear. INTEGRATED SOFT TISSUE and bone surgery may be a viable alternative for patients who have rejected CPAP as a treatment option. Formal sleep testing has shown that surgery and CPAP therapy are equally successful at resolving OSA. REPEAT SLEEP TESTING conducted several years after patients have undergone combined soft tissue and bone surgery has demonstrated consistent and reliable surgical results. Patients with OSA should be offered surgery as an alternative to life-long CPAP treatment or as an option when CPAP therapy has not been successful. PMID- 16309067 TI - Medication safety--reliability of preference cards. AB - A CLINICAL ANALYSIS of surgeons' preference cards was initiated in one hospital as part of a comprehensive analysis to reduce medication-error risks by standardizing and simplifying the intraoperative medication-use process specific to the sterile field. THE PREFERENCE CARD ANALYSIS involved two subanalyses: a review of the information as it appeared on the cards and a failure mode and effects analysis of the process involved in using and maintaining the cards. THE ANALYSIS FOUND that the preference card system in use at this hospital is outdated. Variations and inconsistencies within the preference card system indicate that the use of preference cards as guides for medication selection for surgical procedures presents an opportunity for medication errors to occur. PMID- 16309068 TI - Advanced practice nurse entrepreneurs in a multidisciplinary surgical-assisting partnership. AB - CHANGES IN THE HEALTH CARE environment and reimbursement practices are creating opportunities for nurse entrepreneurs to be partners with other professional nurses and physicians. Advanced practice nurses (APNs) who want to step into an entrepreneurial role must have strong clinical expertise, specific personal characteristics, interpersonal skills, and business acumen. ESTABLISHING A MULTIDISCIPLINARY partnership for providing surgical assisting services has many benefits and presents many challenges. PMID- 16309069 TI - Effects of end-of-life education on baccalaureate nursing students. AB - MOST NURSES MUST DEAL WITH DEATH and dying at some time during their careers; therefore, all nurses should have an understanding of the care people require at the end of life. Many schools of nursing, however, do not require end-of-life issues as a separate subject in the curriculum. THIS ARTICLE DISCUSSES baccalaureate nursing students' experiences during a four-month course in end-of life issues. A PROJECT EVALUATED the change in students' perceived comfort level in dealing with a patient who is dying and the patient's family members. PMID- 16309070 TI - Reducing the risk of cross-contamination from transmissible spongiform encephalopathies. AB - IN RESPONSE TO RECENT REPORTS from several major US medical centers regarding possible exposure of surgical patients to transmissible spongiform encephalopathies (TSEs), one institution developed guidelines to care for surgical patients with the preoperative diagnosis of "rule out TSE." BASED ON CURRENT sterilization technology, using disposable surgical instruments may be the safest way to eliminate concerns of iatrogenic transmission of TSE and reduce the number of hospital staff members who could be exposed to the TSE contagion. PERIOPERATIVE PRECAUTIONS that can be taken when patients are undergoing brain biopsy procedures to rule out TSE are detailed. PMID- 16309071 TI - Technical support assistants--a new concept in the OR. AB - INCREASINGLY COMPLEX TECHNOLOGY, a shortage of surgical first assistants, and the growth of surgical services provided the impetus for expanding the technical support assistant role in one facility. AN EDUCATIONAL PROGRAM was developed based on the job description and the participants' learning needs. PROGRAM EVALUATIONS were positive and reinforced the success of the educational program. PMID- 16309072 TI - AORN standards for RN first assistant education programs. PMID- 16309073 TI - The concept of presencing in perioperative nursing. PMID- 16309074 TI - Implementing National Patient Safety Goal 3. PMID- 16309075 TI - Focus on RN-as-circulator legislation. PMID- 16309076 TI - [Opinion of present development in clinical work of acupuncture and moxibustion]. PMID- 16309077 TI - [Evaluation of clinical therapeutic effects and safety of acupuncture treatment for premenstrual syndrome]. AB - OBJECTIVE: To assess the effectiveness and safety of various types of acupuncture in treatment of premenstrual syndrome (PMS). METHODS: Search Chinese and English databases of the Cochrane Library, CENTRAL, MEDLINE, EMBASE, CBM, CNKI and reference lists of correlative academic conference proceedings. RESULTS: Eight controlled studies involving 807 women (range, 61-225) meeting the enrolled criteria. Among them 7 studies showed that therapeutic effects of acupuncture were superior to other methods (i. e. Western medicine or Chinese herbs). CONCLUSION: Acupuncture can effectively treat PMS. However, more randomized controlled trials are needed to assess the effectiveness and adverse-effect of acupuncture for treatment of PMS. PMID- 16309078 TI - [Multicenter controlled study on transient asthma-stopping action of acupuncture at "Qingchuan point"]. AB - OBJECTIVE: To test and verify the transient therapeutic effect of acupuncture at point "Qingchuan" on bronchial asthma. METHODS: Two hundred cases of bronchial asthma at acute attack stage were divided into a trial group of 100 cases treated with acupuncture at point "Qingchuan" and a control group of 100 cases treated with acupuncture at Dingchuan (EX-B1). RESULTS: The total effective rate was 92.60% and the effect occurred within 42-860 seconds after acupuncture in the trial group, and 81.0% and within 114-126 seconds in the control group, respectively, with very significant differences between the two groups (P < 0.01, P < 0.001). CONCLUSION: Acupuncture at point "Qingchuan" can significantly improve asthmatic state in the patient of bronchial asthma with action of rapidly stopping asthma. PMID- 16309079 TI - [Multicenter controlled study on abdominal acupuncture for treatment of nerve root type cervical spondylosis]. AB - OBJECTIVE: To observe the therapeutic effect of abdominal acupuncture on nerve root cervical spondylosis. METHODS: Three hundred cases were randomly assigned to a treatment group and a traction control group. The treatment group were treated with abdominal acupuncture at an acupoint prescription, i.e. Zhongwan (CV 12), Guanyuan (CV 4) and others were selected. The control group were treated with suspended traction by a cervical vertebral extraction belt at pillow-jaw sitting position. The short-term and long-term therapeutic effects were observed at the end of therapeutic course and 90 days after the end of treatment, respectively. RESULTS: The total short-term and the long-term therapeutic effects were 97.3% and 92.7% in the treatment group, and 82.0% and 82.0% in the control group, respectively, with significant differences between the two groups (P < 0.01). CONCLUSION: Abdominal acupuncture has better short-term and long-term therapeutic effects with short therapeutic course. PMID- 16309080 TI - [Clinical observation on acupuncture combined with music for treatment of Alzheimer disease]. AB - OBJECTIVE: To find the method for increasing clinical therapeutic effect on Alzheimer disease (AD). METHODS: The patients of AD meeting American NINCDS-ADRDA Criteria were randomly divided into group A and group B. The group A (n = 21) were treated with acupuncture for 10 courses, 10 sessions constituting one course; and the group B (n = 20) were treated with combination of acupuncture and music therapy. The acupuncture method was the same as the group A and the music therapy was given for 20 sessions, once every 5 days. After treatment, their effectiveness were evaluated with the Mini-Mental State Examination (MMSE) and Activities of Daily Living (ADL). RESULTS: After treatment, the scores for MMSE increased significantly (P < 0.05) and the scores for ADL decreased significantly (P < 0.05) in the both groups, and the therapeutic effect in the group A was better than the group B (P < 0.05). CONCLUSION: Acupuncture combined with music therapy can increase the therapeutic effect on AD. PMID- 16309081 TI - [Observation on therapeutic effects of different methods on peripheral facial paralysis at different stages]. AB - OBJECTIVE: To observe therapeutic effects of different therapeutic methods for treatment of peripheral facial paralysis at different stages. METHODS: In the treatment group, 26 cases at acute stage were treated by acupuncture at corresponding acupoints at the healthy side, 15 cases at resting stage by acupuncture at the affected side with shallow needling method, and 2 cases at restoration stage by routine acupuncture and moxibustion, and their therapeutic effects were compared with the control group of 36 cases who were treated by routine acupuncture and moxibustion. The acupoints selected in the two groups were same. RESULTS: The cured-markedly effective rate was 97.7% in the treatment group and 75.0% in the control group with a very significant difference between the two groups (P < 0.01). The treatment group needed 2.7 therapeutic courses and the control group needed 4.0 courses in average with a very significant difference between the two groups (P < 0.01). CONCLUSION: Different acupuncture methods at different stages are better than routine acupuncture method in treatment of peripheral facial paralysis. PMID- 16309082 TI - [Clinically controlled study on phased integral needling method for treatment of cerebral infarction]. AB - OBJECTIVE: To compare clinical therapeutic effects of phased integral acupuncture and routine acupuncture on patients with cerebral infarction. METHODS: One hundred and thirteen cases were randomly divided into a treatment group (n = 63) treated by phased integral acupuncture and a control group (n = 50) treated by routine acupuncture. Their clinical therapeutic effects were compared after treatment. RESULTS: The treatment group in the decreases of both the diagnostic score for TCM diseases of stroke and the score for neurological function defect, and the therapeutic effect was superior to the control group with significant differences (all P < 0.05). CONCLUSION: The phased integral acupuncture is an effective therapy with a therapeutic effect better than that of the routine acupuncture for treatment of hemiplegia due to cerebral infarction. PMID- 16309083 TI - [Observation on clinical therapeutic effects of electroacupuncture combined with moving cupping on somatoform disorders]. AB - OBJECTIVE: To observe therapeutic effect of electroacupuncture combined with moving cupping therapy on somatoform disorders. METHODS: Twenty-five cases of somatoform disorders meeting CCMD-3 criteria were treated with electroacupuncture and moving cupping for 4 therapeutic courses (20 sessions), and the improvement of symptoms and therapeutic effect were observed after treatment. RESULTS: After treatment, of the 25 cases, 13 were markedly effective, 9 improved and 3 ineffective, the total effective rate being 88.0%; the score of SCL-90 factor decreased significantly (P < 0.05). CONCLUSION: Electroacupuncture combined with moving cupping therapy can early control target symptoms and improve bad emotion for the patient of somatoform disorders. PMID- 16309084 TI - [Acupuncture treatment of vocal nodule and evaluation of the therapeutic effect by computerized acoustics assay]. AB - OBJECTIVE: To evaluate the therapeutic effect of acupuncture at point Kaiyin No. 1 on vocal nodule. METHODS: Eighty cases of vocal nodule were randomly divided into an acupuncture group of 40 cases, a Chinese herb group of 20 cases and a western medicine group of 20 cases. The acupuncture group were treated with acupuncture at main point Kaiyin No. 1, the Chinese herb group with oral administration of Jinsang Sanjie Pill and the Western medicine group with spray inhalation of antibiotics and hormones. Their therapeutic effects and parameters of acoustics before and after treatment were observed. RESULTS: The total effective rate was 92.5% in the acupuncture group, significantly better than 85.0% in the Chinese herb group and 85.0% in the Western medicine group (P < 0. 01); the acupuncture group in improvement of parameters of acoustics was significantly superior to other two groups (both P < 0.01). CONCLUSION: Acupuncture at point Kaiyin No. 1 has significant therapeutic effect on vocal nodule with no adverse effect. PMID- 16309085 TI - [Effects of acupuncture and continuous multiple points pulse stimulation on linguistic frequency hearing loss in the patient of sensorineural hearing loss]. AB - OBJECTIVE: To explore effective therapeutic methods for sensorineural hearing loss (SNHL). METHODS: Thirty-four cases of mild and moderate SNHL who were definitely diagnosed with pure tone test and by otologists, including 24 cases of insufficiency of the liver and kidney, 10 cases of excess of liver-fire. They were treated with acupuncture at main points, Ermen (TE 21) or Tinggong (SI 19) or Tinghui (GB 2), and Yifeng (TE 17), Shenting (GV 24), Baihui (GB 20), Houding (GV19), Touwei (ST 9), Conger 1-3, Congnao 1-2; and adjuvant acupoints: Huangshu (KI 16), Qihai (CV 6), Guanyuan (CV 4), Taixi (KI 3) were added for insufficiency of the liver and kidney; Zhongwan (CV 12), Tianshu (ST 25), Yanglingquan (GB 34) and Taichong (LR 3) were added for excess of liver-fire. Sequential multipoint pulse stimulation was adopted, twice each week, for 20 consecutive weeks (40 sessions) and then pure tone test was carried out repeatedly. RESULTS: Of 34 cases (58 affected ears), 13 cases were cured and 21 improved, and the various linguistic frequency hearing threshold pure hearing values before and after treatment increased significantly (P < 0.05). CONCLUSION: Acupuncture and continuous multipoint pulse stimulation has definite therapeutic effect on linguistic frequency hearing loss in the patient of SNHL. PMID- 16309086 TI - [Three-dimensional reconstruction research on Mingmen (GV 4)]. AB - OBJECTIVE: To show the space structures of Mingmen (GV 4) area and its adjacent tissues. METHODS: A frozen female corpse was cut into about 0.5 mm thick sections with a high-speed steel plane machine and photographed with digital and optical camera. Two 2-D transverse phase picture data sets were obtained and then were sequenced, matched, modified and divided. The 3-D reconstruction software was used to reconstruct the outline form of the lumbar part containing Mingmen (GV 4) to attain transverse phase picture data sets (3 022 sheets, 0.57 mm in thickness). The sagittal, coronal and different oblique plane were cut and the tissue structures in these pictures were analyzed. Thus, the reconstruction data sets were obtained. CONCLUSION: After the structures of Mingmen (GV 4) were reconstructed with computer, the form of Mingmen (GV 4) can be dynamically observed from any angle and the space construction law of gross form of Mingmen (GV 4) can be proved. PMID- 16309087 TI - [Relation of indications of Back-shu points of zang- and fu-organs with The Bladder Channel of Foot-Taiyang]. AB - OBJECTIVE: To study on relation of indications of Back-shu points of zang- and fu organs with The Bladder Channel of Foot-Taiyang. METHODS: With analysis of origin of Back-shu points of zang- and fu-organs and their indications, the relation of Back-shu points with the incidental and fundamental aspects, theory of qi-flow, and the characteristics of the parts of indications of acupoints reflected by Back-shu points, relations of indications of Back-shu points of zang- and fu organs with The Bladder Channel of Foot-Taiyang were studied. RESULTS: Indications of Back-shu points, a group of acupoints on the circulation route of The Foot-Taiyang Channel, are not found in the channel, which is difficult to explain from twelve channel theory. However, the relation of the incidental and fundamental aspects, theory of qi-flow, and the characteristics of the parts of indications of acupoints with the indications, and the branches of channels presented due to channel tropism of acupoints, etc. contribute to explain the indications of Back-shu points for diseases and syndromes of five zang-organs and six fu-organs. CONCLUSION: The relation of indications of acupoints with disease groups and circulation route of channels can not be discussed simply by Miraculous Pivot . Channels. PMID- 16309088 TI - [Effects of acupuncture at different periods on circadian rhythms of locomotor activity and core body temperature in hamsters]. AB - OBJECTIVE: To explore the time law of electroacupuncture in regulation of circadian rhythms of the organism. METHODS: Effects of electroacupuncture at "Shenshu" (BL 23) at Zi, Wu, Mao and You periods on circadian rhythms of locomotor activity and core body temperature in hamsters were observed with chronobiological research methods. RESULTS: Electroacupuncture at Wu period could decrease the amplitude of locomotor activity rhythm (P < 0.05), at Mao period could delay the peak phase of circadian rhythm and at You period could advance the peak phase of circadian rhythm (both P < 0.05); and electroacupuncture at Mao period could delay 22.36 degrees and at You period advance 39.32 degrees for the rhythm peak of the circadian rhythm of core body temperature. CONCLUSION: Acupuncture has a certain effect on circadian rhythm of locomotor activity and core body temperature. PMID- 16309089 TI - [Effects of manual acupuncture and electroacupuncture on mitochondria of skeletal muscle cells in rats of acute swimming exercise]. AB - OBJECTIVE: To observe effects of acupuncture and electroacupuncture on antioxidant enzyme and Ca2+(-) ATPase activities and Ca2+ content in mitochondria of skeletal muscle cells in rats of acute swimming exercise and explore the mechanism of acupuncture in increasing sport ability. METHODS: Forty male SD rats were randomly divided into 4 groups. The rats in the acupuncture group and the electroacupuncture group were treated respectively with acupuncture and electroacupuncture before acute swimming exercise. After the swimming exercise, the rats were killed and the skeletal muscle was taken, and then the indexes were determined. RESULTS: After acute swimming exercise, the GSH-Px and Ca(2+)-ATPase activities in the mitochondria of the skeletal muscle decreased significantly (P < 0.05) and Ca2+ content increased (P < 0.01) in the swimming group as compared with the control group; in the acupuncture group, SOD activity increased significantly (P < 0.05), GSH-Px and Ca(2+)-ATPase activities increased as compared with the swimming group (P < 0.05); Ca2+ contents in the swimming group and the electroacupuncture group significantly increased as compared with the control group (P < 0.01). CONCLUSION: Acupuncture can protect cells from injury of acute sports and maintain the functions of mitochondria so as to delay fatigue, prolong working time of muscles and prevent muscles from damage. PMID- 16309090 TI - [Comparative observation on therapeutic effects of electroacupuncture and manual acupuncture on female urethral syndrome]. AB - OBJECTIVE: To evaluate objectively clinical effects of different needling methods on urethral syndrome. METHODS: Eighty-nine cases of female urethral syndrome were randomly divided into an electroacupuncture group and a hand-acupuncture group. The scores were evaluated with I-PSS of International Urine-Controlled Association and life quality before and after treatment, and their therapeutic effects were compared. RESULTS: The abnormal symptoms of urination alliviated in the both groups (P < 0.05); the short-term cured rate was 51.8% in the electroacupuncture group, which was higher than 17.1% in the hand-acupuncture group (P < 0.05). CONCLUSION: The therapeutic effect of electroacupuncture on female urethral syndrome is better than that of the hand-acupuncture. PMID- 16309091 TI - [On acupuncture and moxibustion treatment based on the layers of "skin, vessel, muscle, tendon and bone"]. AB - OBJECTIVE: To expound the therapeutic method based on the layers of "skin, vessel, muscle, tendon and bone" and its clinical values. METHODS: Systematically explore the literature relative to treatments based on the layers in The Yellow Emperor's Internal Classic, cite, analyze and summarize contents of nine types of needling and needling methods, etc., relative to treatments based on the layers. RESULTS: The principle and therapeutic methods based on the layers of "skin, vessel, muscle, tendon and bone" are expounded and it is hold that this therapeutic method is one of key theories of ancient acupuncture and moxibustion therapy. The theory of channels and collaterals and treatment based on the layers are two important aspects in acupuncture and moxibustion treatment. Combination of the two aspects can increase therapeutic effect of acupuncture and moxibustion. CONCLUSION: The treatment based on layers of "skin, vessel, muscle, tendon and bone" is of important significance for recognition of acupuncture values in The Yellow Emperor's Internal Classic, a new establishment of diagnosis and examination, syndrome differentiation, treatment system of acupuncture and moxibustion independent of drug prescription, enriching acupuncture and moxibustion therapy, increasing the therapeutic effect of acupuncture and moxibustion, and extending the range of acupuncture and moxibustion treatment. PMID- 16309092 TI - [Su Tian-you's historical relics of practicing medicine and teaching abroad]. AB - OBJECTIVE: To find historical relics of propagation of Chengjiang acupuncture and moxibustion school of thought abroad in which Cheng Dan' an is representative. METHODS: Interview Xing Jingqing, Zhao Zhixing, Qiu Rongqing, students of Ph.D. Su Tianyou in Malaysia, who is the third generation of students of Cheng Dan' an, and collect historical materials about practicing medicine and teaching of Ph.D. Su abroad. RESULTS: Su Tianyou is a student of Zeng Tianzhi, a brilliant disciple of Cheng Dan' an. He practiced medicine in 1939 and established Acupuncture and Moxibustion Medical College of Hong Kong in 1940, and he went to 13 countries and districts such as Asia, America and so on for practicing medicine, running schools. He is respectfully called "father of American acupuncture and moxibustion". CONCLUSION: Ph.D. Su propagated Chinese medicine abroad, with outstanding achievement in education of medical sciences. PMID- 16309093 TI - [Multicenter randomized controlled study on acupuncture-massage comprehensive program for treatment of cervical spondylosis of arterial type]. AB - OBJECTIVE: To probe into clinical value of the acupuncture-massage comprehensive program for treatment of cervical spondylosis of arterial type (CSA). METHODS: By using three-center, single-blind, randomized, controlled method, the eligible subjects were randomly divided into an acupuncture-massage group, an acupuncture group, a massage group, 60 cases in each group. They were treated with acupuncture-massage comprehensive program, simple acupuncture and simple massage, respectively. And multiple-dimensional indexes were established to evaluate the therapeutic effect and safety. RESULTS: The baselines of the general data were no different among the 3 groups. The cured-markedly effective and the effective rate were 68.3% and 88.3% in the acupuncture-massage group, 51.7% and 75.0% in the acupuncture group, and 50.0% and 76.7% in the massage group, respectively, the acupuncture-massage group being better than the other 2 groups (P<0.05); after treatment, the total cumulative scores for symptoms in the 3 groups decreased significantly (all P<0.01), the acupuncture-massage group being superior to the other 2 groups (P<0.05); and the physical signs in the 3 groups improved significantly (P<0.01), with no significant differences among the 3 groups (P>0.05); the time inducing the effect in the acupuncture-massage group was shorter than that in the other 2 groups (P<0.05 or P<0.01); and no adverse effects were found in the 3 groups. CONCLUSION: This acupuncture-massage comprehensive therapy has a definite therapeutic effect on CSA with safety, and it is of superiority as compared with the simple acupuncture or the simple massage. PMID- 16309094 TI - [Clinical observation on hemiplegia early rehabilitation effects of different acupuncture programs]. AB - OBJECTIVE: To observe early rehabilitation of two different acupuncture programs for hemiplegia due to stroke. METHODS: Seventy cases of early hemiplegia due to stroke were randomly divided into a treatment group and a control group. The treatment group were treated by acupuncture at acupoints of both the healthy and affected sides, twice each day, respectively, and the control group by acupuncture at the affected side, once daily. Their therapeutic effects were evaluated by the brief Fugl-Meyer movement scale and the modified Barthel indexes. RESULTS: The patients with over grade IV for FMA accounted for 68.6% and the good rate for ADL was 74.3% in the treatment group, and 31.4% and 42.8% in the control group, respectively, the therapeutic effect of the treatment group being better than that of the control group (P<0.01 or P<0.05). CONCLUSION: The acupuncture program in which acupoints at both the healthy and affected sides are applied alternately is more beneficial to recovery of motor function and activity daily living (ADL) of the patient with hemiplegia due to stroke. PMID- 16309095 TI - [Clinical study on acupoint catgut embedding for treatment of simple obesity]. AB - OBJECTIVE: To explore clinical mechanisms of acupoint catgut embedding for treatment of simple obesity. METHODS: Thirty-two cases of simple obesity were treated by acupoint catgut embedding and acupoints were selected based on differentiation of symptoms and signs. After treatment for 1 or 2 therapeutic courses, the clinical therapeutic effect and changes of the body weight, obesity degree, body mass index, circumferences of the chest, lumbus and buttock. and blood lipids and blood glucose were investigated. RESULTS: After one course the total effective rate was 84.4% and after 2 courses it was 90.0%. After the treatment, the various indexes all changed significantly, the longer the treatment time, the better the therapeutic effect. CONCLUSION: Acupoint catgut embedding has a satisfactory therapeutic effect on obesity, with advantages of simple manipulation and saving time. PMID- 16309096 TI - [Observation on therapeutic effect of scalp acupuncture combined with body acupuncture on stroke]. AB - OBJECTIVE: To search for the best acupuncture and moxibustion treatment program for stroke. METHODS: One hundred and eighty-three cases of stroke were divided into a scalp-body acupuncture group of 62 cases, a scalp acupuncture group of 60 cases and a body acupuncture group of 61 cases. They were treated for 2 months and then their therapeutic effects were compared. RESULTS: In the scalp-body acupuncture group, 13 cases were basically cured, 32 cases improved obviously, 15 cases improved, 2 cases was ineffective, and corresponding figures in the scalp acupuncture group were 8, 21, 24, 7, and in the body acupuncture group were 11, 28, 19, 3, with a significant difference between the scalp-body acupuncture group and the scalp acupuncture group (P<0.05) , and with no significant differences between the scalp-body acupuncture group and body acupuncture group, and between the body acupuncture group and the scalp acupuncture group. CONCLUSION: Scalp acupuncture combined with body acupuncture has a better clinical therapeutic effect than that of simple scalp acupuncture for the patient of stroke. PMID- 16309097 TI - [Acupuncture for treatment of simple obesity and its effect on serum leptin level of the patient]. AB - OBJECTIVE: To explore the therapeutic effect and the mechanism of acupuncture in reducing body weight. METHODS: Fifty cases of simple obesity were randomly divided into an acupuncture group and a drug control group, 25 cases in each group. The patients in the acupuncture group were treated with body acupuncture at Tianshu (ST 25), Guanyuan (CV 4), etc, electroacupuncture and ear point tapping and pressing therapy (Shenmen, Endocrine, etc. were selected); the patients in the control group were treated with oral administration of sibutramine. The serum leptin level before and after treatment were determined and the therapeutic effect in reducing body weight was assessed. RESULTS: The total effective rate was 88.0% in the acupuncture group and 80. 0% in the control group, with no significant difference between the two groups (P>0.05); after treatment, the serum leptin level in both the two groups decreased significantly (P<0.01), and the decreasing amplitude in the acupuncture group was superior to that in the control group (P<0.05). CONCLUSION: Acupuncture therapy has a definite therapeutic effect in reducing body weight, and the improvement of serum leptin level of the patient is possibly one of the mechanisms of acupuncture in reducing body weight. PMID- 16309098 TI - [Clinical observation on shallow needling for treatment of acute intracerebral hemorrhage]. AB - OBJECTIVE: To approach to the best therapy for acute intracerebral hemorrhage (ICH). METHODS: Eighty-five cases of acute ICH were randomly divided into a treatment group (n = 45) and a control group (n = 40). The control group were treated by routine Western medicine, and the treatment group by the medication plus shallow needling. RESULTS: The total effective rate was 95.5% in the treatment group and 45.0% in the control group with a very significant difference between the two groups (P<0.01). CONCLUSION: Shallow needling and the western medicine have cooperation with a better therapeutic effect in treatment of acute intracerebral hemorrhage. PMID- 16309099 TI - [Comparative study on treatment of diabetic gastroparesis by acupuncture and Western medicine]. AB - OBJECTIVE: To compare therapeutic effects of acupuncture and mosapride in relieving diabetic gastroparesis. METHODS: Forty cases were randomly divided into a treatment group and a control group, 20 cases in each group. The treatment group were treated by acupuncture at Zhongwan (CV 12) and Zusanli (ST 36), and the control group by oral administration of mosapride 5 mg, t. i. d. Their therapeutic effects were evaluated by indexes of therapeutic effect. RESULTS: The total effective rate was 95.0% in the treatment group and 83.3% in the control group with a significant difference between the two groups (P<0.05), and the adverse effect in the treatment group was less than that in the control group (P<0.05). CONCLUSION: Acupuncture can significantly relieve diabetic gastroparesis, the therapeutic effect being better than mosapride, with a less adverse effects. PMID- 16309100 TI - [Clinical therapeutic effect of point-injection combined with body acupuncture on prementstrual tension syndrome]. AB - OBJECTIVE: To compare clinical therapeutic effects of acupoint-injection combined with body acupuncture and medication on premenstrual tension syndrome, and probe to the best therapy for this disease. METHODS: One hundred and two cases were randomly divided into an acupoint-injection plus body acupuncture group (treatment group) and a medication group (control group). The treatment group were treated with injection of Huangqi Injection into Zusanli (ST 36), Sanyinjiao (SP 6), and acupuncture at Hegu (LI 4), Taichong (LR 3), Guanyuan (CV 4), Qihai (CV 6), Taiyang (EX-HN5), Fengchi (GB 20) 10 days before menstruation, once every three days; the control group were treated by oral administration of diazepam and oryzanol at routine dose 10 days before menstruation until stopping of menstrual. Their therapeutic effects were compared after treatment for 3 menstrual cycles. RESULTS: The total effective rate was 92.6% in the treatment group and 75.0% in the control group with a significant difference between the two groups (P<0.05). CONCLUSION: The clinical therapeutic effect of the acupoint-injection combined with body acupuncture is better than that of the medication on premenstrual tension syndrome. PMID- 16309101 TI - [Auricular point sticking plus moxibustion for treatment of recurrent herpes simplex]. AB - OBJECTIVE: To search for the best therapy for recurrent herpes simplex. METHODS: Ninety-six cases were randomly divided into an auricular point sticking plus moxibustion group (treatment group, n = 54), a medication group (control group, n = 42). The treatment group were treated with auricular sticking at points Spleen, Lung, Kidney, and others, and moxibustion was applied at Zusanli (ST 36), Fenglong (ST 40), and local blister; the control group were treated by oral administration and external application of aciclovir. RESULTS: The cured rate and the total effective rate were 55.5% and 100.0% in the treatment group, and 9.5% and 61.9% in the control group, respectively, with a significant difference between the two groups (P<0.05). CONCLUSION: The auricular point sticking plus moxibustion is a better therapy for recurrent herpes simplex, with a significantly better therapeutic effect than that of the medication. PMID- 16309102 TI - [To evaluate the effectiveness of a standardized protocol of auricular therapy using magnetic pearls on sleep promotion in the elderly]. AB - OBJECTIVE: To examine the effectiveness of adopting a standardized protocol of auricular therapy using magnetic pearls on sleep promotion in the elderly. METHODS: Sixty eligible participants were invited to receive a 3-week auricular therapy using magnetic pearls (@66 Gauss). Seven auricular points (Shenmen, Heart, Kidney, Liver, Spleen, Subcortex, Occiput) which are thought to have an effect on promoting sleep in the elderly were selected. RESULTS: Significant improvement in the sleep efficiency (SE) collected by wrist actigraphy was observed before, during and after treatment course (P<0.01). The overall effective rate in insomnia with different TCM syndrome types ranged from 66.7% to 90.9% (average 81.7%). No significant difference in the effect of the therapy in terms of SE could be observe between clients with excessive or deficiency syndrome even when the treatment protocol was standardized (P>0.05). CONCLUSION: A standardized protocol of auricular therapy using magnetic pearls for sleep improvement might be appropriate for the ageing population who are homogeneous in terms of age and general health status. PMID- 16309103 TI - [Treatment of knee joint pain with superficial needling]. AB - OBJECTIVE: To compare therapeutic effects of superficial needling and body acupuncture on knee joint pain. METHODS: Ninety-six cases of knee joint pain were randomly divided into a treatment group of 48 cases treated with superficial needling, and a control group of 48 cases treated with body acupuncture. RESULTS: The cured rate was 66.7% in the treatment group and 39.6% in the control group with a significant difference between the two groups (P<0.01). The total effective rate was 100.0% in both the two groups. The needed therapeutic times of superficial needling was significantly less than that of the body acupuncture (P< 0.05). CONCLUSION: Superficial needling has a good therapeutic effect on knee joint pain with a shorter therapeutic course. PMID- 16309104 TI - [Recognition of links of the brain with the channels and collaterals in the Huangdi's Internal Classic]. AB - OBJECTIVE: To approach to recognition of links between the brain and meridians at the early stage of formation of TCM theory. METHODS: The relative papers in the Huangdi's Internal Classic were analyzed, compared, induced and synthesized, attaining the recognition of links between the brain and meridians. RESULTS: In the Huangdi's Internal Classic, it is indicated that the main meridians directly connected with the brain have The Foot-Yangming, The Foot-Taiyang Channels, and The Governor and The Qiao Vessels; the eye system, the vertex, Fengfu (GV 16) and the jaw are the parts of the meridians passing in and out the brain; in the brain there are some pathways of meridians. CONCLUSION: The Huangdi's Internal Classic has richer recognition of the links between the brain and meridians, to which should be pay attention in clinical diagnosis of brain diseases. PMID- 16309105 TI - [Effect of electroacupuncture on metabolism of lipids in rats of obesity induced by sodium glutamate]. AB - OBJECTIVE: To explore mechanisms of acupuncture for reducing of body weight. METHODS: The rat obesity model was developed by subcutaneous injection of sodium glutamate solution, and the experimental rats were divided into a blank control group, a model group, an electroacupuncture group and a sibutramine group; blood TC, TG, HDL-C, LDL-C contents and lipoprotein lipase (LPL) activity, and serum leptin and insulin levels were determined. RESULTS: The body weight and Lee's index, and TG, TC and LDL-C contents in the electroacupuncture group decreased significantly as compared with those in the model group (all P<0.01), the actions in the electroacupuncture group being superior to those in the sibutramine group; the HDL-C contents in both the electroacupuncture and the sibutramine groups were higher than that of the model group (P<0.01) with no significant difference between the electroacupuncture group and the sibutramine group; LPL activity in the electroacupuncture group increased significantly as compared with that in the model group (P<0.01); serum leptin and insulin levels in both the electroacupuncture group and the sibutramine group were lower than that in the model group (P<0.05, P<0.01), with no significant difference between the electroacupuncture group and the sibutramine group in the effect on insulin content, and the decrease of leptin level in the electroacupuncture group was more as compared with the sibutramine group (P<0.01). CONCLUSION: Electroacupuncture can improve the status of high blood lipids, increase lipoprotein lipase activity, and regulate serum leptin and insulin levels in the obestic rat. PMID- 16309106 TI - [Effects of different segments of acupuncture serum on eosinophil counts in the rat with eosinophilia]. AB - OBJECTIVE: To further analyze and identify effective components of anti-asthma in acupuncture serum. METHODS: Changes of eosinophils in the peripheral blood of rats with eosinophilia were observed for 10 days after intravenous injection of the different segments of serum (serum: normal saline = 1:20, 2.5 mL/kg, from the first day of the model establishment, for 3 consecutive days). RESULTS: After intravenous injection of different segments of serum, the eosinophil counts in the peripheral blood decreased significantly from the 3rd day as compared with those of the model group (P<0.05). CONCLUSION: The effective components of acupuncture serum from asthmatic rats treated by acupuncture for eosinophils are not a single component, and acupuncture stimulation may produce many kinds of components of anti-asthma. PMID- 16309107 TI - [Acupuncture and moxibustion in Germany]. AB - Although Germany is an important cradle of Western medicine, traditional Chinese acupuncture and moxibustion are widely accepted and have been developed hardly in Germany for several centuries. This paper reviews the development courses of acupuncture and moxibustion in Germany, and gives an introduction to the level of acupuncture and moxibustion science, acupuncture in medical insurance, acupuncture research, acupuncture clinics, acupuncture organizations, acupuncture journals and trend of acupuncture development in Germany. PMID- 16309108 TI - [Conception of Ashi points]. AB - In the light of obscure conception of Ashi points in the circle of acupuncture and moxibustion at present, this article tries to clarify the origin and definition of Ashi points by textual research of literatures. It is put forward that Ashi points are not the same with "tender spot" and "Buding point, Tianying point", but are some special responding points, including regular points and extra points, when the organism is ill. When these points are pressed the organism will be comfortable or painful. And the definition, location and clinical location method of Ashi points are proposed. PMID- 16309109 TI - [Formation, development and changes of the eye acupunture region-dividing acupoint location program]. AB - Eye acupuncture, a micro-needling therapy, has been widely applied in clinical treatment for over 30 years. Its theoretic system, especially, the eye acupuncture region-dividing point location program has been revised for 2 times, and the third program with reasonability of acupoint distribution, strictness of angle of acupoint region is easily accepted by clinical workers. In the paper, the theoretical basis of professor Peng Jingshan's inspection of the eye for recognition of diseases, diagnosis of diseases and eye acupuncture therapy were explained, and the information, development and change of the eye acupuncture region-dividing acupoint location program were analyzed and the history of development of eye acupuncture were introduced so as to direct clinical application of eye acupuncture. PMID- 16309110 TI - [Advances of studies on treatment of dysmenorrhea with acupoint application]. AB - OBJECTIVE: To summarize advances of studies on treatment of dysmenorrhea with acupoint application in recent 10 years so as to provide references and basis for scientific research and clinical works in future. METHODS: The literature about treatment of dysmenorrhea with acupoint application in the recent 10 years are reviewed from treatment based on syndrome types, acupoint selection, selection of excipients, application time, assessment of therapeutic effects, adverse effects and other aspects. CONCLUSION: Acupoint application has a definite therapeutic effect on dysmenorrhea, but with some problems in clinical research. PMID- 16309111 TI - A chess lesson. PMID- 16309112 TI - The perception of emotion from body movement in point-light displays of interpersonal dialogue. AB - We examined whether it is possible to identify the emotional content of behaviour from point-light displays where pairs of actors are engaged in interpersonal communication. These actors displayed a series of emotions, which included sadness, anger, joy, disgust, fear, and romantic love. In experiment 1, subjects viewed brief clips of these point-light displays presented the right way up and upside down. In experiment 2, the importance of the interaction between the two figures in the recognition of emotion was examined. Subjects were shown upright versions of (i) the original pairs (dyads), (ii) a single actor (monad), and (iii) a dyad comprising a single actor and his/her mirror image (reflected dyad). In each experiment, the subjects rated the emotional content of the displays by moving a slider along a horizontal scale. All of the emotions received a rating for every clip. In experiment 1, when the displays were upright, the correct emotions were identified in each case except disgust; but, when the displays were inverted, performance was significantly diminished for some emotions. In experiment 2, the recognition of love and joy was impaired by the absence of the acting partner, and the recognition of sadness, joy, and fear was impaired in the non-veridical (mirror image) displays. These findings both support and extend previous research by showing that biological motion is sufficient for the perception of emotion, although inversion affects performance. Moreover, emotion perception from biological motion can be affected by the veridical or non veridical social context within the displays. PMID- 16309113 TI - Kanizsa's shrinkage illusion produced by a misapplied 3-D corrective mechanism. AB - In order to include the monocular areas from the left and the right eye in the cyclopean view, the visual system displaces the occluded elements which would result in a horizontal elongation of the shape but does not occur thanks to a correction mechanism which preserves the shape. We hypothesised that this mechanism causes Kanizsa's amodal shrinkage illusion (the apparent elongation of a partially occluded square) when it is incorrectly applied by the visual system to a two-dimensional stimulus. Four experiments tested this hypothesis: (i) one eyed observers were less susceptible to the illusion than people with normal binocular vision because, for them, the correction for shape is unnecessary; (ii) the illusion was stronger with binocular than with monocular vision since binocularity induces the visual system to correct for the shape distortion; (iii) the illusion diminished when the stimulus was rotated 90 degrees given that displacement and compression are not required for vertical occlusion; (iv) the magnitude of the illusion was a function of the width of the occluder because, as previous research has shown, the edges of a partially occluded square are less displaced the farther they are from the edges of the occluder. The data from the four experiments support our hypothesis even though no condition was able to eliminate the illusion; other possible causes are discussed. PMID- 16309114 TI - Colour, polarity, disparity, and texture contributions to motion segregation. AB - We measured how different cues are combined in motion-segregation processes by using motion stimuli where randomly distributed target dots were organised in global revolving motion while the remaining noise dots performed random motion. Target dots were cued with a different colour, polarity, disparity depth, or texture orientation than the noise dots, or they were the same as the noise dots. The stimuli were presented with a prolonged static cue preview which provided position cues to target dots or, briefly with static pre-target and post-target noise frames, which provided false position cues (no preview). All cues efficiently facilitated global motion segregation in cued-preview conditions. Colour completely failed to facilitate global motion segregation in no-preview conditions. Polarity and disparity facilitated segregation in no-preview conditions, although sensitivities were lower than in the preview conditions. Remarkably, texture orientation largely facilitated motion segregation by the same amount in both cued-preview and no-preview conditions. So, colour provides only position cues to the motion-segregation task whereas texture orientation, disparity, and to a lesser extent polarity are integrated with the segregation process. PMID- 16309115 TI - Temporal properties of disparity processing revealed by dynamic random-dot stereograms. AB - In studies of the temporal flexibility of the stereoscopic system, it has been suggested that two different processes of binocular depth perception could be responsible for the flexibility: tolerance for interocular delays and temporal integration of correlation. None has investigated the relationship between tolerance for delays and temporal integration mechanisms and none has revealed which mechanism is responsible for depth perception in dynamic random-dot stereograms. We address these questions in the present study. Across five experiments, we investigated the temporal properties of stereopsis by varying interocular correlation as a function of time in controlled ways. We presented different types of dynamic random-dot stereograms, each consisting of two pairs of alternating random-dot patterns. Our experimental results demonstrate that (i) disparities from simultaneous monocular inputs dominate those from interocular delayed inputs; (ii) stereopsis is limited by temporal properties of monocular luminance mechanisms; and (iii) depth perception in dynamic random-dot stereograms results from cross-correlation-like operation on two simultaneous monocular inputs that represent the retinal images after having been subjected to a process of monocular temporal integration of luminance. PMID- 16309116 TI - Children's search behaviour in large-scale space: developmental components of exploration. AB - It has been argued that visual-search tasks provide a valid model for foraging behaviour. However, Gilchrist et al (2001 Perception 30 1459-1464) demonstrated that, whilst some aspects of behaviour transferred to large-scale egocentric search, there were substantially fewer revisits to previously searched locations than would be expected from the visual-search literature. This difference might be a result of the greater effort required to search in a large-scale egocentric context. Here, we present a novel, automated paradigm, for examining the effect of effort on egocentric search behaviour by manipulating the motor difficulty of the task. Children searched for a hidden target amongst a randomised display of lights by activating a switch at each potential location. The motor difficulty of the task was manipulated by requiring children to search with either their dominant or their nondominant hand. We found that when children searched with their nondominant hand, they made significantly more revisits to previously checked locations than they did when using their dominant hand. This suggests that, when the motor response was more effortful, children were less able to efficiently guide their search behaviour. Individuals with a greater visuo spatial short-term memory span performed the task more quickly than those with a lower span. However, search latencies were unrelated to general fluid intelligence. This highlights the role of spatial working memory in the development of efficient exploration of large-scale space. PMID- 16309117 TI - Human spatial navigation via a visuo-tactile sensory substitution system. AB - Spatial navigation within a real 3-D maze was investigated to study space perception on the sole basis of tactile information transmitted by means of a 'tactile vision substitution system' (TVSS) allowing the conversion of optical images-collected by a micro camera-into 'tactile images' via a matrix in contact with the skin. The development of such a device is based on concepts of cerebral and functional plasticity, enabling subjective reproduction of visual images from tactile data processing. Blindfolded sighted subjects had to remotely control the movements of a robot on which the TVSS camera was mounted. Once familiarised with the cues in the maze, the subjects were given two exploration sessions. Performance was analysed according to an objective point of view (exploration time, discrimination capacity), as well as a subjective one (speech). The task was successfully carried out from the very first session. As the subjects took a different path during each navigation, a gradual improvement in performance (discrimination and exploration time) was noted, generating a phenomenon of learning. Moreover, subjective analysis revealed an evolution of the spatialisation process towards distal attribution. Finally, some emotional expressions seemed to reflect the genesis of 'qualia' (emotional qualities of stimulation). PMID- 16309118 TI - Spatial modulation of tactile temporal-order judgments. AB - We report a series of three experiments designed to examine the effect of posture on tactile temporal processing. Observers reported which of two tactile stimuli, presented to the left and right index fingers (experiments 1-3; or thumb, experiment 3), was perceived first while adopting one of two postures--hands close (adjacent, but not touching) or hands-far (1 m apart)--in the dark. Just noticeable differences were significantly smaller in the hands-far posture across all three experiments. In the first two experiments we compared hand versus foot responses and found equivalent advantages for the hands-far posture. In the final experiment the stimuli were presented to either the same or different digit on each hand (index finger or thumb) and we found that only when the same digit on each hand was stimulated was there an advantage for the hands-far posture. The finding that temporal precision was better with greater distance contradicts predictions based on attention-switching models of temporal-order judgments, and also contrasts with results from similar experimental manipulations in other modalities (eg vision). These results provide support for a rapid and automatic process that transforms the representation of a tactile stimulus from a skin centred reference frame to a more external (eg body-centred or allocentric) one. PMID- 16309119 TI - Depth and motion in historical descriptions of motion parallax. AB - Motion parallax was described as a cue to depth over 300 years ago and as producing apparent motion over 150 years ago. In recent years, experimental interest in motion parallax has increased, following the rediscovery of the idea that stimulus motion can be yoked to head movement. We compare the historical descriptions with some contemporary research, which indicates how depth and motion perception are dependent on the conditions of stimulation. PMID- 16309120 TI - Sound and sight: acoustic figures and visual phenomena. AB - The impact that the analysis of sound waves has made on theories of light is well known, and is touched upon here. However, the acoustic figures described initially by Robert Hooke in 1665 and in more detail by Ernst Chladni in 1787 (often referred to as Chladni figures) were instrumental in vision in two specific respects. First, their representation by Tyndall [1867 Sound. A Course of Eight Lectures Delivered at the Royal Institution of Great Britain (London: Longmans, Green)] in a book on sound resulted in the description of a visual illusion, the Hermann grid. Secondly, attempts to render the acoustic figures visible (on the basis of briefly persisting images) led to the discovery of instruments that could synthesise movement. These two developments are discussed in their historical contexts. PMID- 16309121 TI - The identification and perception of bottled water. PMID- 16309122 TI - [Role of nitric oxide in the mechanisms of formation of reflex vasomotor responses]. AB - This review focuses on modem data concerning the role of nitric oxide (NO) in the mechanisms of vasomotor regulation. On the background of the literature data and own experimental results, we have discussed some questions of NO integration into transmission of impulses in central and autonomic nervous system under condition of realization of cardiogenic and sinocarotid pressor and depressor reflexes, reflectory vasomotor responses formation under acute myocardial ischemia. According to literature and own functional and morphological data we suggest species differences in NO participations in mechanisms of reflex self--regulation of circulation. PMID- 16309123 TI - [Dominating motivation in systemic memory mechanisms]. AB - The materials provided in the article support the key role of dominating motivation in the systemic processes of fixation and opening of memory mechanisms. The activating mechanisms of dominating motivations in the systemic architectonics of behavioural acts provide the basis for development of a multicomponent acceptor apparatus of an action outcomes broadly represented in various analysing brain sections. As result of enhancement of action outcomes on acceptors structures, molecular behaviour engrammes form within the functional systems. It is these molecular engrammes that are opened by dominating motivations in the same spatial-temporal sequence in which training takes place, and determine deliberate actions of animals. It was demonstrated that dominating motivation opens genetic information with an approximating-exploratory reaction under strong activation of early genes expression, in particular, of c-fos gene protein. Inherent motivation reactions are not blocked by inhibitors of proteins synthesis, by cycloheximide, in particular. In the process of training animals, i.e., satisfaction of the demands which are the basis of dominating motivations, expression of early genes in reduced, while expression of late genes is initiated. In this case, blockators of protein synthesis begin to produce strong inhibiting impact on behaviour of animals. PMID- 16309124 TI - [A psychophysiological approach toward the problem of consciousness]. AB - Psychophysiological analysis of the relationship between the level of consciousness and the brain activation permitted an identification of three levels of consciousness at the transition from wakefulness to sleep. A change in the intensity and quality of involuntarily mental processes served as an index of the level of consciousness according to subjects' self-reports. It was found that a certain EEG profile corresponds to each level of consciousness under study. Data obtained showed that the lowest level of activation corresponds to a state in which a subject notes an inhibition of "internal speech", "failure of thoughts". At this case the dynamics of the EEG high frequency rhythm changes which is possibly related to the mechanism of transmission of cognitive information. PMID- 16309125 TI - [The electrophysiologycal and information aspects of stress development]. AB - In the first part of this paper the modern concepts of stress and possibility of diagnostics of early stress manifestations are considered. Though significant advances of researches of this problem many methodological and methodical aspects are continued to be discussed. The categorical proff are stage development of adaptation processes and there classification [H. Selye]. A long-term studies of adaptation processes of students in semesters and exams have been conducted by authors with specially development and to accumulate new data. The second part of this paper is the survey of these results published in different time. The polyparametric technology including some electrophysiological methods with the system analysis using artificial intelligence algorithms and methods of pattern recognition permitted to show some regular dependences in relationships of electrophysiological parameters and there variations under stress development. It was shown for diagnostics of stress stages in necessary to assess not only absolute values of parameters but also there relationships, which are new diagnostics signs considering as information connections between physiological systems. The main characteristic of functional state of an organism is balance of parameter's relationships in totality, representing invariant for healthy persons. In condition of disharmony of parameter relationships there are aroused the strain, overstrain functional state or failure of adaptation in depends on a degree of misbalance. The represented new data don't contradict to known reasons and mechanisms of stress and accumulated data of its variety manifestations. As the work-hypothesis the information-wave model of appearance of psychosomatic reactions of stress is introduced. PMID- 16309126 TI - [I.P. Razenkov and his achievements (50-th anniversary of death)]. PMID- 16309127 TI - [Clinical application of acupuncture for treatment of heroin withdrawal syndrome]. AB - OBJECTIVE: To make technical standard of acupuncture manipulation for acupuncture treatment of heroin withdrawal syndrome. METHODS: Two hundred and twenty cases of heroin withdrawal syndrome were randomly divided into an acupuncture group of 111 cases and a control group of 109 cases. They were respectively treated with acupuncture and oral administration of lofexidine hydrochloride, and their therapeutic effects were observed. RESULTS: The heroin dependence (acute stage) were effectively withdrawn in the two groups. The treatment group in change of total scores for withdrawal symptoms before and after treatment, the total scores for withdrawal symptoms at the 4th and 5th days, treatment of insomnia and the score for self-Hamilton Anxiety Scale and the score after at the 4th day was superior to the control group (P < 0.05, P < 0.01, P < 0.001). CONCLUSION: Acupuncture has a satisfactory, rapid, safe and reliable clinical therapeutic effect. PMID- 16309128 TI - [Clinical observation on needle pricking for treatment of infertility due to varicocele after varicocelectomy]. AB - OBJECTIVE: To observe clinical therapeutic effect of needle pricking therapy on infertility with varicocele after varicocelectomy. METHODS: One hundred and twenty-two cases were randomly divided into a treatment group of 62 cases and a control group of 60 cases. The treatment group were treated with needle pricking at sacral plexus nerve spots and lumbar plexus nerve spots, and the control group were treated with intramuscular injection of HCG. RESULTS: After treatment, the pregnancy rate of the patient's wife was 79.0% in the treatment group and 41.7% in the control group with a significant difference between the two groups (P < 0.01); the production hormones improved significantly in the both groups (P < 0.01); superoxide dismutase and Zn in the semen elevated significantly in the treatment group (P < 0.05). CONCLUSION: Needle pricking therapy can significantly improve and regulate endocrines in the patient of varicocele after varicocelectomy, and elevate quality of semen and sperm and the pregnancy rate of the patient's wife. PMID- 16309129 TI - [Observation on therapeutic effect of acupuncture combined with drug for treatment of intestinal metaplasia of chronic atrophic gastritis]. AB - OBJECTIVE: To search for an effective method for treatment of intestinal metaplasia of gastric mucosa in the patient of chronic atrophic gastritis. METHODS: Fifty-one cases of chronic atrophic gastritis with intestinal metaplasia were randomly divided into a treatment group (n = 26) and a control group (n = 25). The treatment group were treated with acupuncture at main acupoints: Zhongwan (CV 12), Zusanli (ST 36), Neiguan (PC 6), Gongsun (SP 4), and oral administration of modified Sijunzi Decoction, and the control group with the modified Sijunzi Decoction. After treatment for 3 months, clinical symptoms, results of gastroscopy and pathological changes of gastric mucosa were compared between the two groups. RESULTS: There was no significant difference in improvement of the clinical symptoms between the two groups (P > 0.05); the total effective rate of 88.46% for gastroscopy in the treatment group was better than 64.00% in the control group (P < 0.05); and the total effective rate of 84.62% for pathological improvement of intestinal metaplasia in the treatment group was superior to 64.00% in the control group (P < 0.05). CONCLUSION: Acupuncture combined with Chinese drug is a better therapy for intestinal metaplasia of gastric mucosa in the patient of chronic atrophic gastritis. PMID- 16309130 TI - [Effect of integral syndrome differentiation acupuncture on the tear film stability in the patient of xerophthalmia]. AB - OBJECTIVE: To probe into a treatment method for increasing clinical therapeutic effect on xerophthalmia. METHODS: Forty-five cases were randomly divided into acupuncture group 1, acupuncture group 2 and control group. The acupuncture group 1 were treated with acupuncture based on syndrome types, and the acupuncture group 2 with acupuncture at local acupoints, and the control group with dripping of artificial tears. Local symptoms of eyes, S1T, BUT and FL were observed before and after treatment. RESULTS: S1T and BUT increased after treatment in the two groups (P < 0.01, P < 0.05), but did not significantly change in the control group (P > 0.05). There were significant differences in change of FL as the control group compared with the acupuncture group 1 and 2. The total effective rate was 86.7%, 66.7% and 33.3% in the acupuncture group 1, 2 and the control group, respectively. CONCLUSION: The integral syndrome differentiation acupuncture can improve secretion of tear, increase stability of tear film, improve local symptoms of eyes with a therapeutic effect better than acupuncture at local acupoints and artificial tears. PMID- 16309131 TI - [Controlled study of needle warming therapy and electroacupuncture on simple obesity of spleen deficiency type]. AB - OBJECTIVE: To compare therapeutic effects of needle warming therapy and electroacupuncture on simple obesity with spleen deficiency. METHODS: Sixty-eight cases of simple obesity with spleen deficiency, including water-dampness retention due to spleen deficiency, qi-deficiency of the lung and spleen, yang deficiency of the spleen and kidney, were randomly divided into a needle warming therapy group (n = 36) and an electroacupuncture group (n = 32). Zhongwan(CV 12), Shuifen (CV 9), Qihai (CV 6), Zhongji (CV 3), Tianshu (ST 25), Shuidao (ST 28), and so on were selected as main acupoints in the both groups. Their therapeutic effects and body mass indexes were observed. RESULTS: The total effective rate was 88.9% in the needle warming therapy group and 71.9% in the electroacupuncture group with a significant difference between the two groups (P < 0.05); and there was a significant difference between the two groups in decrease of body mass index (P < 0.05). CONCLUSION: Needle warming therapy has definite and long-term therapeutic effect on simple obesity with spleen deficiency. PMID- 16309132 TI - [Observation on therapeutic effect of acupuncture and moxibustion on hyperemesis gravidarum]. AB - OBJECTIVE: To search for the best method for treatment of hyperemesis gravidarum. METHODS: One hundred and fifty cases of hyperemesis gravidarum were randomly divided into 3 groups, acup-moxibustion group, Chinese drug group and Western medicine group, 50 cases in each group. The acup-moxibustion group were treated with acupuncture at Zhongwan (CV 12), Neiguan (PC 6), Zusanli (ST 36) and Yinlingquan (SP 9), and after arrival of qi, mild warming moxibustion of 10-15 min was added; the Chinese drug group with oral administration of modified Suye Huanglian Decoction , and the Western medicine group with oral administration of phenobarbital and supplement of water and electrolyte. RESULTS: The total effective rate was 86.0% in the acup-moxibustion group, 42.0% in the Chinese drug group and 38.0% in the Western medicine group with a significant difference among the 3 groups (P < 0.05), the therapeutic effect in the acup-moxibustion group being the best. CONCLUSION: Acup-moxibustion is the best method for hyperemesis gravidarum. PMID- 16309133 TI - [Clinical observation on abdominal cluster-needling combined with sacro-iliac needling for treatment of chronic pelvic inflammation]. AB - OBJECTIVE: To observe therapeutic effect of abdominal cluster-needling combined with sacro-iliac-needling on chronic pelvic inflammation. METHODS: One hundred and ten cases of pelvic inflammation were randomly divided into a treatment group and a control group. The treatment group of 70 cases were treated by abdominal cluster-needling combined with sacro-iliac-needling; the control group of 40 cases were treated by oral administration of Fuyankang tablet. Their therapeutic effects were compared. RESULTS: The cured rate and the total effective rate were 75.7% and 97.1% in the treatment group, and 37.5% and 85.0% in the control group, respectively, with significant difference between the two groups (P < 0.01, P < 0.05). CONCLUSION: The therapeutic effect of abdominal cluster-needling combined with sacro-iliac-needling is significantly better than that in the control group for chronic pelvic inflammation. PMID- 16309134 TI - [Analysis on blood-pressure-decreasing effect of electric pulse stimulation of auricular acupoint Jiangyagou in the patient of hypertension]. AB - OBJECTIVE: To investigate the blood-pressure-decreasing action of stimulating auricular acupoint Jiangyagou in the patient of hypertension. METHODS: One hundred and twenty-nine cases of stage I - III hypertension were treated by electric pulse stimulation of auricular acupoint Jiangyagou with a MEDI/calma Nerve and Muscle Stimulator, 20-30 min each time, once daily. After treatment of 3 courses (21 days), changes of blood pressure were observed. RESULTS: Of the 129 cases, 44 cases were markedly effective, 73 cases were effective and 12 cases were ineffective, the total effective rate being 90.70%. The systolic pressure was decreased by (18.64 +/- 1.48)mmHg and the diastolic pressure by (8.01 +/- 0.68)mmHg on average, with very significant differences before and after treatment in both the systolic pressure and the diastolic pressure. CONCLUSION: Electric pulse stimulation of Jiangyagou can effectively decrease both the systolic pressure and diastolic pressure in the patient of hypertension. PMID- 16309135 TI - [Comparison of the thermal conductivity of the related tissues along the meridian and the non-meridian]. AB - OBJECTIVE: To compare the thermal conductivity of the related tissues along meridian line and non-meridian area. METHODS: Forty healthy volunteers were observed with a infrared thermal imaging system. RESULTS: During heating the acupoint or the non-acupoint along the meridian line, the velocity of spreading of the temperature-increasing response along the meridian line was more easy and rapid, with a definite direction. However, during heating the non-meridian spots, the change of temperature was confined to the local area, with no definite direction. The spreading of skin temperature response along the meridian line was more rapid than the non-meridian area during both the two were heated simultaneously, and finally, 3 infrared radiant tracks along the meridian courses conformed basically with that of the classical three yin- meridians of the hand appeared. CONCLUSION: The thermal conductivity of the related tissues along the meridian line is better than that of non-meridian area and has definite direction, with differences in physical characteristics between them. PMID- 16309136 TI - [On functional area of Back-shu based on relationship between Back-shu points and Jiaji points]. AB - OBJECTIVE: To prove the existence of functional area of Back-shu and provide a new thinking for clinical practice. METHODS: The consistency between the Back-shu points and Jiaji points at the same level of spinal column was discussed from the origin and development, mechanisms and clinical application of Back-shu and Jiaji points. CONCLUSION: The points at the same level of spinal column has same origin of development, mechanisms and clinical functions, so as to prove the existence of functional area of Back-shu. PMID- 16309137 TI - [Observation on therapeutic effect of "Shandong Xiaoying" needling method for treatment of cerebral infarction]. AB - OBJECTIVE: To search for an effective method for treatment of cerebral infarction. METHODS: Sixty cases of cerebral infarction were randomly divided into an observation group and a control group, 30 cases in each group. All of them were treated with routine needling method, and "Shandong Xiaoying" needling was added to the observation group. The therapeutic effects, recovery of myodynamia and daily living ability were compared in the two groups. RESULTS: There was no significant difference between the two groups in the total effective rate for recovery of upper limb myodynamia (P > 0.05), but there was a very significant difference in the total effective rate for recovery of lower limb myodynamia and increase of daily living ability between the two groups (P < 0.01). CONCLUSION: "Shandong Xiaoying" needling has markedly therapeutic effect on cerebral infarction. PMID- 16309138 TI - [Preliminary study on Professor SUN Liu-he's academic thought]. AB - Professor SUN Liu-he has been engaged in scientific research, clinical and teaching works for tens years and he is a meticulous scholar with great learning, rich clinical experience and great theoretical attainments. In clinical practice of many years, he summarizes and develops Kangai Plaster, Kangai Fushui Plaster, Ai tong Ling Plaster, and so on for acupoint sticking to treat tumors, and establishes Nu-needle hot-reinforcing method and Ti-Yun reducing method, enriching contents of needling methods, and he is good at application of specific points, established points and channel acupoints to treat refractory diseases with good results. PMID- 16309139 TI - [Relation of left and right axillary temperature with Yin-Yang and its guiding action on diagnosis and treatment of acupuncture and moxibustion]. AB - Body temperature is one of the four physical signs and clinically, the axillary temperature is the most commonly used as the index of body temperature. The study on relation of the difference of the left and right axillary temperature with constitution types and diagnosis of diseases proves that the temperature of the left and right axillary is correlated with excess and insufficiency of Yin and Yang, namely, higher left axillary temperature is recognized as Yin-deficiency and Yang-excess, and higher right axillary temperature as Yang-deficiency and Yin excess, and this can guide clinical syndrome differentiation treatment of acupuncture and moxibustion and individual diagnostic and therapeutic program can be established based on this. PMID- 16309140 TI - [Discussion on several commonly-seen problems in teaching of the Acupuncture and Moxibustion Science]. AB - The arrange of teaching material, finger force in needling, rapidly needle inserting method, heavy thrusting and light lifting the needle, light thrusting and heavy lifting the needle, keeping qi, needling depth and other aspects are discussed in combination with experiences in teaching of The Acupuncture and Moxibustion Science and clinical practice of many years, so as to help to grasp relative knowledge for beginners in learning Acupuncture and Moxibustion Science. PMID- 16309141 TI - [Study on LUO Zhao-ju's academic thought in the new Chinese Acup-Mox Surgical Therapeutics]. AB - OBJECTIVE: To probe preliminarily academic thought of New Chinese Acup-Mox Surgical Therapeutics written by LUO Zhao-ju. METHODS: Read, analyze and study New Chinese Acup-Mox Surgical Therapeutics written by LUO Zhao-ju, and sum up the academic thought. CONCLUSION: The main academic thought and values of the book embody in the seven aspects: Spreading and popularizing acupuncture and moxibustion medicine; developing surgery of acupuncture and moxibustion; elucidating cause of disease and pathogenesis; advocating treatment in accordance with different symptoms and signs; paying great attention to the furuncle; highly praising the simple and practical prescriptions; initiating acupuncture combined with medication of Chinese drugs. PMID- 16309142 TI - [Experimental study on action of acupoint embedding thread on ANCA in the rat of ulcerative colitis]. AB - OBJECTIVE: To observe therapeutic effect of acupoint embedding thread therapy on ulcerative colitis and to study the mechanism. METHODS: The rat model of ulcerative colitis was developed with 2,4-dinitrochlorobenzene method. The 20th day after the end of model development, they were randomly divided into a model group, a salicylazosulfapyridine (SASP) treatment group and an acupoint embedding thread therapy group. The SASP treatment group were treated with intragastrical perfusion of 4.5% SASP suspension, and the acupoint embedding thread therapy group were treated with embedding thread at "Zusanli" (ST 36) and "Shangjuxu" (ST 37). Anti-neutrophil cytoplasmic antibody (ANCA) was detected with indirect immunofluorescence, and the pathological changes were investigated with pathological tissue section technique. RESULTS: The positive rate of ANCA reached to 62.5% in the rat with ulcerative colitis with significant differences among the 3 groups. After treatment, the positive rate of ANCA was decreased in the two treatment groups (P < 0.05, P < 0.01). CONCLUSION: Acupoint embedding thread therapy has definite therapeutic effect on ulcerative colitis and the mechanism is possibly related with the decrease of ANCA. PMID- 16309143 TI - [Effect of electroacupuncture on expression of IFN-gamma-like immunoreactive substance in spinal cord of the rat ]. AB - OBJECTIVE: To observe effect of electroacupuncture at "Zusanli" (ST 36) on expression of interferon-gamma (IFN-gamma)-like immunoreactive substance in spinal cord of the rat and to probe the mechanism. METHODS: The IFN-gamma-like immunoreactive positive cell number in spinal cord of the rat was investigated with immunohistochemical SP method and microscopy. RESULTS: After electroacupuncture at "Zusanli" (ST 36), IFN-gamma-like immunoreactive positive cell number in the spinal cord of the rat with electroacupuncture plus immunosuppression did not significantly change (P > 0.05). The number of positive cells in the dorsal horn in the rats with immunosuppression was significantly less than that in the normal control group (P < 0.05) with no significant change in other parts and with no significant difference between the electroacupuncture group and the control group (P > 0.05). CONCLUSION: Electroacupuncture at "Zusanli" (ST 36) can increase expression of IFN-gamma-like immunoreactive substance in spinal cord of the rat, and acupuncture activates the nerve immunoregulative network possibly by IFN-gamma as medium. PMID- 16309144 TI - [Advances of clinical studies on acupuncture and moxibustion for treatment of obesity in recent 10 years]. AB - OBJECTIVE: To Sum up advances of studies on acupuncture for treatment of obesity in recent 10 years and to put forward the problems should be paid attention to and the development direction in clinical scientific research. METHODS: Review the literature about acupuncture and moxibustion slimming therapy in recent 10 years from characteristics of acupuncture and moxibustion in selection of acupoints, analysis of various therapeutic methods, problems in clinical research. CONCLUSION: Acupuncture and moxibustion treatment of obesity has obvious therapeutic effect and less side-effects with flexible selection of acupoints and more methods. PMID- 16309145 TI - [A systematic evaluation of randomized controlled trials for acupuncture and moxibustion treatment of insomnia]. AB - OBJECTIVE: To assess the therapeutic effect of acupuncture on insomnia. METHODS: A systematic evaluation of all relevant randomized controlled trials (RCT) about acupuncture and moxibustion treatment of insomnia was carried out. The data were statistically analyzed with a special software RevMan 4.1. RESULTS: Six papers of RCT including 585 patients met the enrolled criteria. All of the trials were of lower in methodological quality without using blind method; Meta-analysis indicated that odds ratio was 3.55, 95% confidence interval [2.50, 5.05], P < 0.0001, as acupuncture group compared with drug group. CONCLUSION: Acupuncture is possibly effective for insomnia. However, because of lower methodological quality of all the trials, this conclusion has no enough evidence. PMID- 16309146 TI - [Study on clinical therapeutic effect of activating brain and regaining consciousness needling method on poststroke depression and the mechanism]. AB - OBJECTIVE: To probe into an effective method for treatment of poststroke depression. METHODS: Two hundred and fifty-six cases of poststroke depression were randomly divided into a control group (n = 76) treated by oral administration of amitilin and an acupuncture group (n = 180) treated by activating brain and regaining consciousness needling method. Their clinical therapeutic effects and changes of relative factors of poststroke depression and plasma contents of monoamine neurotransmitters were observed. RESULTS: In the acupuncture group, the clinical symptoms and the relative indexes in the depression scales improved and plasma contents of the monoamine neurotransmitters increased significanty as compared with the control group (P < 0.05). CONCLUSION: The needling method for activating brain and regaining consciousness can improve relative factors to cure poststroke depression. PMID- 16309147 TI - [Clinical observation and mechanism study on acupuncture combined with massage for treatment of vertebrobasilar insufficiency]. AB - OBJECTIVE: To search for the best program increasing clinical therapeutic effect on vertebrobasilar insufficiency (VBI) and to study the mechanism. METHODS: The patients were randomly divided into an acupuncture plus massage group, an acupuncture group and a massage group. Their therapeutic effects were observed and compared. And transcranial Doppler (TCD) was used for detection of systolic flow velocity (Vs), enddiastolic peak flow velocity (Vd), mean peak flow (Vm) and pulsatility index (PI) before and after treatment. RESULTS: The cured rate and the total effective rate in the acupuncture plus massage group were significantly higher than the acupuncture group and the massage group (P < 0.05). And there were significant differences in Vm, PI and DI in the left vertebral artery (LVA), the right vertebral artery (RVA) and the basilar artery (BA) before and after treatment (P < 0.01). CONCLUSION: Acupuncture combined with massage can improve Vm, PI, DI of VA-BA and the clinical therapeutic effect on VBI as well. It is superior to the simple acupuncture therapy and simple massage therapy. At present, it is one of the best therapy for VBI. PMID- 16309148 TI - [Clinical study on scalp acupuncture combined with sports therapy for rehabilitation of poststroke hemiplegia]. AB - OBJECTIVE: To probe into effect of scalp acupuncture combined with sports therapy on movement function of limbs in the patient of poststroke hemiplegia. METHODS: One hundred and twenty cases of poststroke hemiplegia were randomly divided into an observation group and a control group, 60 cases in each group. The observation group were treated by scalp acupuncture and the control group by simple sports therapy. The movement function of limbs were assessed by Fugl-Meyer rating method and Barthel index. RESULTS: The movement function of limbs and ability of daily life after treatment in the observation group improved very significantly as compared with the control group (P < 0.01). CONCLUSION: Scalp acupuncture combined with sports therapy can promote recovery of the movement function of limbs in the patient of poststroke hemiplegia. PMID- 16309149 TI - [Clinical observation on acupoint-injection of Chinese drugs for treatment of hypertensive renal lesion]. AB - OBJECTIVE: To compare therapeutic effects of acupoint-injection of Chinese drug and oral administration of Western medicine on hypertensive renal lesion. METHODS: Fifty-seven cases were randomly divided into two groups. The treatment group of 30 cases were treated by injection of Yuxingcao injectio and Danshen injectio into Shenshu (BL 23) and Zusanli (ST 36), and the control group of 27 cases by nifepine. Their therapeutic effects were compared. RESULTS: The markedly effective rate of 46.7% and the total effective rate of 86.7% in the treatment group were significantly higher than 22.2% and 51.8% in the control group (P < 0.05, P < 0.01). There were significant differences in 24h-urinary protein, urinary beta2-microglobulin (beta2-MG) and N-acetyl-beta-D-glucosaminidase (NAG) after treatment between the two groups (P < 0.01). CONCLUSION: The therapeutic effect of acupoint-injection of Chinese drugs is better than that of nifepine and it has protective action on renal function. PMID- 16309150 TI - [Analysis on therapeutic effect of acupoint-injection on chronic hepatitis B]. AB - OBJECTIVE: To search for an effective method for treatment of chronic hepatitis B. METHODS: One hundred and twenty-three cases were randomly divided into a treatment group (n = 63) and a control group (n = 60). The treatment group were treated with injection of Huangqi injectio and Danshen Injectio into Ganshu (BL 18) and Zusanli (ST 36), once every other day; and the control group were treated with oral administration of Gankangning tablet and fufang yiganling tablet. The clinical symptoms, hepatic function, hepatitis B e antigen (HBeAg), hepatitis B virus-desoxyribose nucleic acid (HBV-DNA) were compared between the two groups. RESULTS: The total effective rate was 93.7% in the treatment group and 76.7% in the control group with significant difference between the two groups (P < 0.01); and the treatment group in hepatic function and the effects of turning negative for HBeAg and HBV-DNA was better than the control group (P < 0.01). CONCLUSION: Acupoint-injection has a better therapeutic effect on chronic hepatitis B. PMID- 16309151 TI - [Observation on therapeutic effect of combination of acupuncture with drug on depression]. AB - OBJECTIVE: To assess clinical therapeutic effect and safety of acupuncture combined with fuxiting on depression. METHODS: Fifty-three cases of depression were divided into an observation group (n = 30) treated by acupuncture combined with fuxiting and a control group treated by fuxiting. Their clinical therapeutic effects were assessed by HAMD, HAMA and CGI, and adverse effects by TESS. RESULTS: After treatment of 6 weeks, the total effective rate was 80.0% in the observation group and 69.6% in the control group with no significant difference between the two groups (P > 0.05). There were significant differences in scores of HAMD and HAMA before and after treatment in the two groups (P < 0.01), with no significant difference between the two groups (P > 0.05). The adverse effect in the observation group was less and milder than that in the control group. CONCLUSION: Acupuncture combined fuxiting has good therapeutic effect on depression with less and mild adverse effects. PMID- 16309152 TI - [Clinical study on analgesic effect of Huaisanzhen on the nerve root pain due to prolapse of lumbar intervertebral disc]. AB - OBJECTIVE: To search for an effective therapy for the nerve root pain caused by prolapse of lumbar intervertebral disc. METHODS: One hundred and ninety-two cases were randomly divided into a Huaisanzhen group of 96 cases, a drug control group of 48 cases and an acupuncture control group of 48 cases. The Huaisanzhen group were treated by Huaisanzhen therapy, the drug control group by intramuscular injection of aspirin-DL-lysine and the acupuncture control group by routine acupuncture. RESULTS: The time inducing analgesic effect was shorter, the effect lasting time was longer, and the analgesic effect and the comprehensive therapeutic effect were better in the treatment group as compared with the two control groups with very significant differences (P < 0.01). CONCLUSION: Huaisanzhen therapy has a better analgesic effect on the nerve root pain due to prolapse of lumbar intervertebral disc. PMID- 16309153 TI - [Observation on therapeutic effect of acupuncture plus salt-partitioned moxibustion at Shenque (CV 8) in 60 cases of chloasma]. AB - OBJECTIVE: To observe the therapeutic effect of acupuncture plus salt-partitioned moxibustion at Shenque (CV 8) on chloasma. METHODS: A total 106 cases of chloasma were randomly divided into two groups, a treatment group (n = 60) treated by acupuncture plus salt-partitioned moxibustion at Shenque (CV 8) and a control group (n = 40) by simple acupuncture. Their therapeutic effects were observed after 3 therapeutic courses and 1-month break, respectively. RESULTS: The total effective rate was 100.0% in the treatment group and 89.1% in the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: Acupuncture plus salt-partitioned moxibustion at Shenque (CV 8) is a safe and convenient therapy with a remarkable effect for chloasma. PMID- 16309154 TI - [Therapeutic effect of acupoint-injection combined with electroacupuncture on chronic pharyngitis]. AB - OBJECTIVE: To compare therapeutic effects of acupoint-injection combined with electroacupuncture and oral administration of medicine on chronic pharyngitis. METHODS: The treatment group of 71 cases were treated by injection of fufangdanshen injectio into Tiantu (CV 22) combined with electroacupuncture at Renying (ST 9), and the control group of 51 cases were treated by oral administration of acetylspiramycin and qingyan tablet. RESULTS: The cured rate and the total effective rate were 60.5% and 90.1% in the treatment group, and 21.6% and 68.6% in the control group, with significantly differences between the two groups (P < 0.01). CONCLUSION: Therapeutic effect of acupoint-injection combined with electroacupuncture on chronic pharyngitis is superior to that of oral administration of medicine. PMID- 16309155 TI - [Analysis on literature about syndrome differentiation and treatment of meridians in clinical acupuncture and moxibustion in recent 3 years]. AB - OBJECTIVE: To understand clinical application of syndrome differentiation and treatment of meridians in acupuncture and moxibustion. METHODS: Collect and collate the literature about syndrome differentiation and treatment of meridians in Chinese Acupuncture & Moxibustion, Acupuncture Research, Shanghai Journal of Acupuncture & Moxibustion (Clinic edition B in 2003) and Clinical Journal of Acupuncture and Moxibustion from 2001 to 2003. RESULTS: Less literature about syndrome differentiation and treatment of meridians are involved, only accounting for 5.69% (163/2 864); and the literature about both treatment based on meridians and treatment based on syndrome differentiation of meridians only accounted for 1.22% (35/2 864). CONCLUSION: At present, clinically acupuncture and moxibustion do not play attention to treatment based on syndrome differentiation of meridians. PMID- 16309156 TI - [Application of Cheng's needling method like a dragonfly skimming the surface of the water in treatment of pain syndrome]. AB - Cheng's needling method like a dragonfly skimming the surface of the water is a needling method handed down in the family of chief TCM physician Cheng Zi-jun and it has been applied clinically for over one hundred years. It is a unique reinforcing-reducing method of acupuncture with simple and convenient manipulation and excellent therapeutic effect, which is created and gradually completed by Cheng's grandfather Mr. Cheng Jin-he on the basis of studying ancient various reinforcing-reducing methods. In the method, stimulating amount of acupuncture is introduced into reinforcing-reducing. Clinically, this method is used to treat various chronic pain syndromes with rapid effect. PMID- 16309157 TI - [On effects of acupoints and drugs in acupoint-injection treatment]. AB - Acupoint-injection therapy is a successful sample for clinical application of integrated Chinese and Western medicine, and has widely been applied clinically. In this method, effects of meridians, acupoints and drugs are organically combined, with obviously higher total therapeutic effect compared with routine acupuncture and moxibustion, intermuscular injection or intravenous injection. In this paper, the total therapeutic effects of acupoint-injection, high effectiveness of drugs and integrated action of acupoint and drug were discussed mainly and it is considered that the basic reason of acupoint-injection producing the high effect of drug is due to meridians and acupoints integrating some pharmacological actions of drugs. PMID- 16309158 TI - [Analysis on ebb-flow of ying-qi]. AB - OBJECTIVE: To analyze and evaluate practical significance of the ebb-flow theory of ying-qi. METHODS: Based on The Yellow Emperior's Internal Classic, the comparisons of the number of channels, collaterals and their bone-length measurement participating in ebb-flow of ying-qi, ying-qi ebb-flow theory with the practically respiratory-pulse ratio, human heart rate and pulse rate, human respiratory times, ying-qi ebb-flow theory with early channel theory, ying-qi ebb flow theory with degrees of whole day, were tested and verified. CONCLUSION: Practically, the ying-qi ebb-flow theory conforms few to human body, which possibly is an assumption of the ancients. PMID- 16309159 TI - [Study on theory and clinical application of meridians (III)]. AB - OBJECTIVE: To probe into essence of meridians. METHODS: To examine meridians, acupoints, brain and relative organs by SQUID and fMRI, and to analyze the results. CONCLUSION: A relative stable circular current of electromagnetic and chemical oscillation along the low electric resistance pathway was found. Competition of different frequency oscillation often yield resonance in some positions of the human body so as to form "oscillatory network" and "strange point" with the electromagnetic and chemical oscillation circulating dominant position in the body, which is possibly meridians and acupoints with regulative actions. The brain is the center of meridians. In different physiological or pathological process, characteristic changes and negatively correlative phenomenon of electromagnetic spectrum at acupoints and the brain are possibly key mechanisms in dynamic regulation of meridians. PMID- 16309160 TI - [Effects of electroacupuncture at neiguan (PC 6) and Shenmen (HT 7) on brain functional imaging]. AB - OBJECTIVE: To observe effect of electroacupuncture at Shenmen (HT 7) and Neiguan (PC 6) on brain functional imaging. METHODS: The technique of functional magnetic resonance imaging was used to observe the activated state in different brain regions caused by electroacupuncture. RESULTS: The frontal lobe was activated by electroacupuncture at Neiguan (PC 6) and the temporal lobe by Shenmen (HT 7). CONCLUSION: Electroacupuncture at different acupoints can activate different brain regions, which provides objective basis for treatment of intellectual impairment by electroacupuncture at Neiguan (PC 6) and Shenmen (HT 7). PMID- 16309161 TI - [Effects of cake-separated moxibustion on ultrastructures of endothelial cells of aorta in the rabbit of hyperlipemia]. AB - OBJECTIVE: To explore the mechanism of cake-separated moxibustion in treatment of hyperlipemia. METHODS: Sixty rabbits were randomly divided into 4 groups, a blank group,a model group, a direct moxibustion group and a cake-separated moxibustion group. Hyperlipemia model was developed by high fat diet of cholesterol. Changes of ultrastructures of endothelial cells of the aorta of the rabbit were observed with electron microscope. RESULTS: The endothelial cells in the cake-separated moxibustion group were more intact, most of them were normal in forms, internal elastic membrane was continuous, their thickness was even, the cells of smooth muscles in the medial membrane were relatively normal, which are similar to those in the blank control group. But the structure of endothelial cells of the aorta in the model group disappeared, in cytoplasm the sedimentation of a great number of lipids can be seen, internal elastic membrane was interrupted, the thickness was uneven, with focal dissolution, the cells of smooth muscle in the medial membrane had sedimentation of lipids, with frothy change. CONCLUSION: Cake separated moxibustion has a certain protective action on endothelial cells of the aorta in the rabbit of hyperlipemia. PMID- 16309162 TI - [Analgesic effects of electroacupuncture of different pulse waveforms in the rat of adjuvant arthritis]. AB - OBJECTIVE: To explore the best pulse waveform of electroacupuncture for treatment of adjuvant arthritis (AA). METHODS: Forty Wistar rats were randomly divided into 5 groups. The electroacupuncture groups of AA were treated with different pulse waveforms of electroacupuncture, respectively. Pain threshold, swelling degrees of joint, and contents of 5-HT, beta-EP and LEK in local tissues of inflammation were observed. RESULTS: After the treatment, pain thresholds of the electroacupuncture groups with different pulse waveforms were higher than that of the model group (P < 0.05, P < 0.01); the local swelling degrees of inflammation in the electroacupuncture group were higher than that in the normal group (P < 0. 01), but was lower than that in the model group (P < 0.05, P < 0. 01); the contents of 5-HT in local tissues of inflammation in the electroacupuncture groups were lower than that in the model group (P < 0.05, P < 0.01); in comparison with the model group, the electroacupuncture of sound-electric wave increased the contents of beta-EP and LEK (P < 0.01, P < 0.05) and the electroacupuncture of intermittent wave increased the content of beta-EP in local tissues of inflammation (P < 0.05). CONCLUSION: Sound-electric wave is the best pulse waveform of electroacupuncture for treatment of adjuvant arthritis. PMID- 16309163 TI - [The point of view of moxibustion in the book, A General Survey of Moxibustion]. AB - OBJECTIVE: To study on academic point of view of the book, A General Survey of Moribustion. METHODS: Study academic thought of the book and clarify academic origin and characteristics of the book. CONCLUSION: Processing of argyi leaf, manufacture of mugwort floss and size of moxa cone have a certain technique standards in the book. The book indicates that small cones should be used with more times of moxibustion. and the properties of mugwort floss are not dry and heat, but are warm and moist and mild, moxibustion not only can be used to treat cold syndrome but also can be used to treat heat syndrome; the body can response to the external moxibustion. The above points of view have still guiding significance in nowadays. PMID- 16309164 TI - Increased levels of PPARbeta/delta and cyclin D1 in flat dysplastic ACF and adenomas in Apc(Min/+) mice. AB - BACKGROUND: In Apc(Min/+) (Min; multiple intestinal neoplasia) mice two separate populations of aberrant crypt foci (ACF) develop in the colon after azoxymethane (AOM) exposure. ACF(Min), with a flat appearance, severe dysplasia and increased beta-catenin expression, are related to adenoma development, whereas classic ACF, with elevated structure, hyperplasia and normal beta-catenin level, are probably not. MATERIALS AND METHODS: The expressions of peroxisome proliferator-activated receptors (PPARs) beta/delta, cyclin D1 and beta-catenin in ACF, adenoma and normal tissue from AOM-treated Apc(Min/+) mice and a familial adenomatous polyposis (FAP) patient colon tumour were assessed by immunohistochemistry and immunoblotting. RESULTS: The flat ACF (ACF(Min)) displayed increased cytoplasmic levels of beta-catenin, and increased levels of cyclin D1 and PPARbeta/delta. In contrast, the expression in classic ACF resembled normal mucosa. Adenomas from Apc(Min/+) mice, as well as a FAP patient colon tumour, displayed increased nuclear and cytoplasmic levels of beta-catenin, and the same expression patterns of cyclin D1 and PPARbeta/delta as those found in flat ACF. CONCLUSION: In addition to activation of the Wnt signalling pathway in both flat ACF and in adenomas in Apc(Min/+) mice, the increased expression of PPARbeta/delta in these lesions could be a target for pro-inflammatory signals important for growth and reduced apoptosis. PMID- 16309165 TI - A novel human monoclonal antibody derived from tumor-infiltrating lymphocytes in lung cancer inhibits cancer cell growth with morphological changes. AB - Tumor-infiltrating lymphocytes (TIL) were isolated from surgically resected human samples using hybridoma techniques, and human monoclonal antibodies (HuMoAbs) were produced. Stable antibody-producing hybridoma cell lines were established and, on the basis of reactivity to human cancer cell lines, a clone of HuMoAb, named HoAKs-1, was selected. By confocal microscopy, we confirmed that HoAKs-1 showed specific and intense reactivity to the cell membrane of HLC-1 and PANC-1, wheareas the antibody did not show reactivity to human umbilical vein endothelial cells (HUVECs). Using xenografts formed from epithelial cancer cell lines in nude mice, we also demonstrated a broad spectrum of reactivity of the antibody in 6 out of 14 xenografts. In addition, using surgically resected clinical specimens from two patients with lung cancer, we showed that HoAKs-1 had specific reactivity to cancerous lesions but not to normal sites. In an in vitro experiment, HoAKs-1 induced morphological changes with neurite-like cytoplasmic processes in MKN-45, HLC-1 and PANC-1, whereas no morphological alterations were observed in HUVECs. Growth inhibition rates by HoAKs-1 were 63% in HLC-1 and 47% in MKN-45, while no growth inhibition occurred in normal HUVECs. We confirmed that HoAKs-1 recognized a 55 kDa protein by determining the molecular weight of the HoAKs-1 reacting protein. In conclusion, we successfully produced a novel HuMoAb, HoAKs-1, which reacted specifically to cancer cells and inhibited cell growth with morphological changes. HoAKs-1 may have the potential to be utilized as an anticancer agent, without causing any immunological reaction in humans. PMID- 16309166 TI - Tumor endothelial cells are targets for selective therapies: in vitro and in vivo models to evaluate antiangiogenic strategies. AB - Angiogenesis is a complicated process, essential for tumor progression and metastasis. Extensive work has been done to understand the mechanisms of tumor angiogenesis and identify angiogenesis inhibitors. It is now recognised that tumor endothelial cells present different functional and phenotypic characteristics than normal resting endothelial cells. These differences and advances in molecular biology have allowed the development of selective agents targeting tumor endothelial cells as therapeutic approaches for cancer. These new targeted strategies need to be evaluated in relevant models before being transferred from the laboratory bench to the clinic. In vivo tumor models remain a good way to evaluate the effect of these agents on tumor growth and metastasis. Nevertheless, in parallel to the development of tumor angiogenesis inhibitors, in vitro models have been designed to mimic angiogenesis steps and enable the evaluation of these new drugs. In this paper, after reviewing the phenotypic characteristics of tumor endothelial cells that make them easy to target for antiangiogenic therapy, some of the most commonly used in vitro and in vivo models, which enable the evaluation of antiangiogenic agents, are presented and discussed. PMID- 16309167 TI - Osteoprotegerin is expressed in colon carcinoma cells. AB - BACKGROUND: Osteoprotegerin (OPG), a soluble member of the tumor necrosis factor family, is produced by various cell types and tissues and plays a key role in the physiological regulation of osteoclast differentiation and activity. Also, OPG is a soluble decoy receptor for tumor necrosis factor-related apoptosis-inducing factor (TRAIL). In the present study we investigated whether the human colon cancer cell lines HT-29 and SW-480 produce and secrete OPG in vitro. MATERIALS AND METHODS: Expression of OPG mRNA was examined by RT-PCR. OPG protein was analysed by ELISA assay and immunostaining methods. The effect of OPG secretion on TRAIL-mediated apoptosis was also investigated. RESULTS: By RT-PCR, it was demonstrated that mRNA transcripts for OPG were produced by both cell lines. By ELISA analysis, OPG was detected in the culture medium; and treatment of cells with proinflammatory cytokines TNF-alpha and IL-1beta increased OPG secretion significantly. Tumor xenografts in nude mice also were shown to express OPG by immunohistochemistry. When RANKL, which selectively binds OPG, was added to cell cultures along with recombinant TRAIL, apoptosis was shown to increase significantly. CONCLUSION: These data indicate that OPG may be involved in tumorigenesis and the progression of colon cancer. PMID- 16309168 TI - Humanized monoclonal antibody against parathyroid hormone-related protein suppresses osteolytic bone metastasis of human breast cancer cells derived from MDA-MB-231. AB - BACKGROUND: Parathyroid hormone-related protein (PTHrP) has been implicated in bone metastasis. However, the effects on bone metastasis of blocking the PTHrP function have not been tested in the clinic. Here, the effects of a humanized anti-PTHrP monoclonal antibody (mAb) on bone metastasis in a human xenograft model are shown. MATERIALS AND METHODS: Subline MDA-5a, with high bone metastatic activity, was established from the human breast cancer cell line MDA-MB-231. Mice were injected with MDA-5a and an anti-PTHrP monoclonal antibody (mAb) raised against human PTHrP (1-34); bone metastasis was evaluated by X-ray photography. RESULTS: MDA-5a produced elevated levels of PTHrP, Interleukin 8 (IL-8), IL-6 and matrix metalloproteinase 1 (MMP-1) and frequently metastasized to the bone. Administration of the humanized anti-PTHrP mAb significantly suppressed osteolytic bone metastasis of MDA-5a and caused osteogenesis at the sites of metastasis. CONCLUSION: The humanized anti-PTHrP mAb was effective against bone metastasis by inducing osteogenesis and, therefore, will provide a new treatment option for bone metastasis in breast cancer. PMID- 16309169 TI - Expression of p27(KIP1) and cell proliferation in human retina and retinoblastoma. AB - BACKGROUND: Retinoblastoma is a rare cancer of the eye, in which biallelic inactivation of the retinoblastoma gene is a hallmark. Although retinoblastoma protein (Rb) and p27(KIP1) block the cell cycle transition from G1- to S-phase, the interaction has not been confirmed in vivo. The aim of this study was to examine the correlation between the expression of p27(KIP1) and cell proliferation in human retina and retinoblastoma. MATERIALS AND METHODS: Human retinoblastoma, surgically removed, was fixed by 4% paraformaldehyde. Then, paraffin-embedded tissue sections were examined using immunohistochemistry with anti-p27(KIP1) and anti-proliferating cell nuclear antigen (PCNA) antibodies. RESULTS: Retinoblastoma tissue was adjacent to the normal retina in which tumor cells with homogeneous nuclei proliferated and it was impossible to identify the layer structure of the inner nuclear layer (INL) and the outer nuclear layer (ONL). In normal retina, PCNA-positive nuclei were not observed, whereas nuclear immunoreactivity for PCNA was detected in a variety of tumor cells. Many p27(KIP1)-positive nuclei were detected in INL and ONL, while p27(KIP1) immunoreactivity was not detected in retinoblastoma cells. CONCLUSION: The correlation between disappearance of p27(KIP1) and induction of proliferation activity suggests that functional loss of Rb leads to down-regulation of p27(KIP1) and uncontrolled retinal cell proliferation. PMID- 16309170 TI - Possible inhibition of cancer cell adhesion to the extracellular matrix in NK4 induced suppression of peritoneal implantation. AB - Milky spots (MS), peritoneal lymphoid tissues, expose the extracellular matrix (ECM) due to a defect of mesothelial cells on their surface, which may explain why peritoneal implantation of cancer cells preferentially takes place at MS. We recently reported that adenovirus vector-mediated intraperitoneal production of NK4 strongly suppressed MS-selective implantation of cancer cells and subsequent peritoneal dissemination, without histological evidence of angiogenesis inhibition. The present study was conducted to clarify the mechanisms underlying the suppressive effects of NK4 on peritoneal implantation. In mice intraperitoneally injected with CT26 cells that were genetically modified to produce NK4 (CT26-NK4), peritoneal dissemination was significantly suppressed with survival prolongation. A decreased cell implantation to omental MS was also detected and evaluated by green fluorescence protein (GFP) imaging. In an in vitro adhesion assay, hepatocyte growth factor-stimulated adhesion to ECM components, such as fibronectin and collagen, was inhibited in CT26-NK4 compared to control cells. These results strongly suggest an inhibition of cancer cell adhesion to the ECM in the suppression of peritoneal implantation by NK4. PMID- 16309171 TI - Morphological, immunocytochemical and flow cytometric in vitro characterisation of a surface-adherent medulloblastoma. AB - Well-characterised cell lines derived from paediatric intrinsic brain tumours are rare. The different repertoire of cell adhesion molecules expressed by primitive neuro-ectodermal tumours, when compared with gliomas, results in a general lack of propensity for surface adherence. In this study, a highly cellular, medulloblastoma biopsy with a Ki-67 index of 20%, obtained by posterior fossa craniotomy of a two-year-old boy, was maintained in surface- adherent culture for twelve sequential in vitro passages. The culture (VC312R) was characterised by immunocytochemistry and flow cytometry using antibodies against cluster of differentiation 44 (CD44), glialfibrillary acidic protein (GFAP), intermediate filament proteins (Nestin and Vimentin), neural cell adhesion molecules (NCAMs) (ERIC and UJ13A), ganglioside (GD3) and neuron-glial 2 (NG2). GD3, GFAP, ERIC-1, UJ13A and NG2 were detected by neither immunocytochemistry nor flow cytometry. It is of particular interest that we have previously reported that the progenitor cell-associated NG2 heparan sulphate proteoglycan was not expressed in a series of medulloblastoma biopsy sections in our laboratories, while NG2 positivity was seen in supratentorial primitive neuro-ectodermal tumours (PNETs). Strong CD44 positivity was detected on most cells (mean = 93.5% of cells on flow cytometry). In one previous case of medulloblastoma, maintained in our laboratories (IPNN-8) as a substrate-adherent culture, no CD44 staining was detected. Twenty-five percent of cells were strongly Vimentin-positive while 54.5% of cells showed Nestin positivity. The expression of Nestin, Vimentin and CD44 is consistent with primitive neural cell evolution. Non-expression of NCAMs may be consistent with the lack of cell-cell adhesion in this culture, which results in surface adherence. The high expression of CD44 may also indicate a distinct phenotype within primitive neuroectodermal tumours, which determines cell-cell and cell extra cellular matrix adhesive properties. PMID- 16309172 TI - Combretastatin A-4 resistance in H460 human lung carcinoma demonstrates distinctive alterations in beta-tubulin isotype expression. AB - Tubulin isotype distribution may play a role in the development of anti-cancer anti-tubulin drug resistance as well as in drug efficacy and specificity. Stepwise selection was used to establish non-small cell lung carcinoma (NSCLC) H460 cells resistant to combretastatin A-4 (CA4), paclitaxel or vinblastine. The results demonstrated that the rate of CA4 drug resistance development was slower than that for paclitaxel. Western analysis demonstrated alterations in total beta tubulin and classes I, III and IV tubulin isotypes among the resistant H460 cell lines. Class III beta-tubulin was significantly altered in all resistant cell lines. Cells resistant to paclitaxel, a structural stabilizer of microtubules, exhibited an increased expression while cells resistant to CA-4 and vinblastine, structural destabilizers of tubulin, demonstrated a reduction of the same isotype. To our knowledge, this is the first demonstration of resistance development and of the corresponding tubulin isotype response for the combretastatins. PMID- 16309173 TI - The photoreceptor protector zeaxanthin induces cell death in neuroblastoma cells. AB - BACKGROUND: The dietary carotenoid zeaxanthin protects against age-related eye disease by preventing apoptosis in photoreceptor cells. This study examined the effect of zeaxanthin on neuroblastoma cells in which apoptosis can be induced with lipid peroxidation products. Since zeaxanthin can inhibit lipid peroxidation and beta-carotene inhibits lipoxygenase (LOX) activity, it was of concern that zeaxanthin might inhibit apoptosis in these cancer cells. MATERIALS AND METHODS: Apoptosis-resistant CHP100 neuroblastoma cells were treated with zeaxanthin. Apoptosis was assessed via an immunoassay for histone-associated DNA fragments and cytofluorimetric analysis of apoptotic body formation. The effect of zeaxanthin on the activity of two model LOXs and LOX-mediated lipid peroxidation in liposomes was assessed. RESULTS: Zeaxanthin strongly induced apoptosis in neuroblastoma cells. Consistent with this finding, zeaxanthin did not inhibit LOX activity. CONCLUSION: Zeaxanthin is a remarkable dietary factor that is able to induce apoptosis in neuroblastoma cells while being able to prevent apoptosis in healthy cells. PMID- 16309174 TI - FGF receptor phosphotyrosine 766 is a target for Grb14 to inhibit MDA-MB-231 human breast cancer cell signaling. AB - BACKGROUND: Fibroblast growth factors receptors (FGFRs) are involved in estrogen independent breast cancer cell growth. Grbl4, a member of the Grb7 family of adapters, is an inhibitor of FGFR signaling. MATERIALS AND METHODS: FGFR from highly invasive MDA-MB-231 cells were expressed in Xenopus oocyte, a widely used model system to question cascade transduction regulations. The effect of microinjection of Grb14 and various mimetic peptides for FGFR tyrosine residues were analysed by FGFR immunoprecipitation and Western blot analysis of signaling cascades. RESULTS: PLCy, ERK2, JNK1 and AKT were blocked by Grb14. Only the pY766 phosphopeptide mimetic of the PLCgamma binding site on FGFR released the inhibitory action of Grb14. CONCLUSION: Grb14 binds to the Y766 site of MDA-MB 231-FGFR, competing for PLCy activation, thus inducing an arrest of the signaling transduction cascades. PMID- 16309175 TI - Scanning electron microscopy of colonic lesions in 1,2-dimethylhydrazine-treated rats. AB - The surface morphology of late colonic lesions in F344 rats treated with 1,2 dimethylhydrazine was studied by scanning electron microscopy. At week 31 after carcinogen treatment, the surface epithelial characteristics of different types of lesions observed in the colonic mucosa were compared, namely classic elevated aberrant crypt foci (ACF), flat lesion and gross tumour. Classic elevated ACF were easily observed as structures with enlarged crypts elevated from the background mucosa. When the various ACF were compared, or when the ACF were compared with the background mucosa, no ultrastructural differences, or differences in the density of goblet cells were found. The flat lesion showed an epithelium without goblet cells and crypts with small openings harbouring a large number of loose, undefined, dysplastic epithelial cells. These changes appeared to be linked to the malignant development since they were also characteristic of the examined tumour. PMID- 16309176 TI - Antitumor effect of double immunization of mice with mucin 1 and its coding DNA. AB - OBJECTIVE: To evaluate the antitumor effect of mouse immunization with human mucin 1 gene (muc 1) DNA plasmids combined with simultaneous injections of human mucin 1 (MUC1) protein. MATERIALS AND METHODS: MUC1 DNA was cloned in pBK-CMV to prepare DNA plasmids and in pET22b(+) to produce proteins. Three strains of mice, immunized with DNA or DNA plus MUC1, were inoculated with tumor cells obtained from spontaneous tumors. IgG(2a) production, MUC1-specific IFN-delta-producing CD8+ T cells, tumor growth and mouse survival were monitored. RESULTS: Only immunization with DNA plus proteins induced IgG(2a) and intracellular IFN-delta production by CD8+ T cells in the strains tested. DNA plus protein immunization induced a better mouse survival in comparison with the DNA groups. However, all immunized mice invariably developed tumors. CONCLUSION: Immunization with DNA plus proteins induced a better protection from tumor growth than immunization with naked DNA. However, the efficacy of immunization with MUC1-based antigens remains low. PMID- 16309177 TI - Preclinical studies of the novel vascular disrupting agent MN-029. AB - BACKGROUND: Vascular disrupting agents (VDAs) are designed to cause a rapid and selective shutdown of the established tumor vasculature, which leads to secondary ischemic tumor cell death. MATERIALS AND METHODS: We examined the efficacy of a novel VDA, MN-029, in the rodent KHT sarcoma model. RESULTS: A significant reduction in the functional vessel number was observed after intraperitoneal injection of MN-029 at a dose of 100 mg/kg. Histological evaluation showed extensive necrosis (approximately 90%) by 24 h. MN-029 treatment to the tumor bearing mice also resulted in a dose-dependent tumor cell killing. When used in combination with radiation or cisplatin chemotherapy, a 100 mg/kg dose of MN-029 significantly enhanced tumor killing compared to that seen with radiation or cisplatin alone. CONCLUSION: The results demonstrated that MN-029 could cause rapid vascular shutdown in solid tumors, dose-dependent secondary tumor cell killing, and effective enhancement of the antitumor effects of radiation and cisplatin chemotherapy. PMID- 16309178 TI - Liposomal transfection of squamous carcinoma cells of the head and neck with IL-2 and B7 plasmids inducing an autologous immune response in vitro. AB - New treatment strategies need to be developed to face the increasing incidence and mortality of squamous cell carcinoma of the head and neck (SCCHN), as the overall survival rate remains poor, with minor therapeutic progress having been achieved over the past forty years. One major goal could be to restore a damaged immune system by intratumoral injection of IL-2-genes that permanently provide non-toxic IL-2-protein concentrations at the tumor site, sufficient to activate cellular immunity in vivo. We showed that the transfection of SCCHN cell lines with IL-2-plasmids, encapsulated in DOTMA/Col, in vitro resulted in the synthesis of bioactive IL-2-protein for up to 30 days by the tumor cells themselves. The transcription of secondary cytokines (IL-6, IL-8, GM-CSF, TNF-alpha) and the expression of immunomodulatory surface molecules (MHC Class II, ICAM1) were enhanced. The IL-2-modified tumor cells were effectively lysed by autologous peripheral blood lymphocytes (PBLs). The immune response was enhanced by B7.1 gene-cotransfection and/or preactivation of PBLs with exogenous IL-2. We demonstrated that in vitro liposome-mediated IL-2-gene-transfection of SCCHN cells is an effective method to stimulate an autologous immune response and is, therefore, promising for clinical application. PMID- 16309179 TI - Differential phosphorylation patterns of P-glycoprotein reconstituted into a proteoliposome system: insight into additional unconventional phosphorylation sites. AB - Membrane vesicles from the multidrug-resistant KB-V1 and KB-C1 cell lines overexpressing P-glycoprotein (Pgp), responsible for pleiotropic chemotherapeutic agents resistance, were solubilized with octyl-glucoside (OG-EX) and further fractionated on DEAE-sepharose column with increased concentrations of NaCl. The fraction containing Pgp (F3) was reconstituted into proteoliposomes (F3-PLP). Comparisons of the phosphorylation levels of Pgp achieved throughout the purification and reconstitution steps were addressed in this study. The [delta32 P] ATP-driven phosphorylation of Pgp was strongly increased in OG-EX, decreased in F3 and not detected in F3-PLP, when compared to Pgp phosphorylation in native plasma membrane vesicles. [delta32 P]ATP-phosphorylation of Pgp in F3-PLP could be restored by exogenously added PKC or by the catalytic sub-unit of PKA. The vanadate-induced hyperphosphorylation effect on Pgp by [delta32 P]ATP observed with plasma membrane vesicles was maintained in OG-EX, but was lost in F3 and did not enable labelling in F3-PLP. Enhancement of [delta32 P]-labelling of native Pgp via [delta32 P]ATP combined with GTP was maintained and also triggered phosphorylation of purified/reconstituted Pgp in F3-PLP as well. Altogether, our data suggest differential phosphorylation patterns of the transporter linked to environmental molecular composition (lipids, presence of detergent) and structure (unfolded versus embedded). In addition, restoration by GTP of Pgp phosphorylation by [delta32 P]ATP in the frame of F3-PLP suggests intra-molecular modulations and hints that other phosphorylation sites and processes, different from the classic ones involving PKC and/or PKA, may participate in the transporter's mechanism. PMID- 16309180 TI - Reverse transcriptase-PCR analysis of apoptosis-regulating gene expression in human benign prostatic hyperplasia. AB - BACKGROUND: To determine whether an imbalanced interaction between proapototic and antiapoptotic signals may account for the loss of the normal cell growth control in benign prostatic hyperplasia (BPH), the expression of some apoptosis regulating genes (bcl-2, bax, c-myc, fas) was investigated. PATIENTS AND METHODS: BPH specimens were obtained from 20 patients who underwent trans-urethral resection of the prostate (TURP) or adenomectomy. Gene expression was studied by reverse transcriptase-polymerase chain reaction (RT-PCR) and its correlation with age and serum PSA level was also investigated. RESULTS: Genes were found to be differentially expressed in BPH tissues. In particular, the antiapoptotic gene bcl-2, which was found in 18/20 samples, gave the weakest signal (p < 0.05 - p < 0.001, Wilcoxon's signed rank test), whereas the cell cycle regulator c-myc was detected in all the specimens and was the most highly expressed (p < 0.001). A positive relationship between the expression of bcl-2 and that of the two proapoptotic genes bax and fas was observed (p < 0.05, Spearman's rank correlation test), as well as between c-myc and fas levels (p < 0.005). Moreover, bax expression positively correlated with age and PSA (p < 0.02), which have also been shown to directly correlate (p < 0.01). CONCLUSION: The higher expression of the oncogene c-myc suggests the activation of mitogenic signals within hyperplastic prostate tissue which a relatively high expression of the proapoptotic genes bax and fas fails to counterbalance. PMID- 16309181 TI - Conjugated linoleic acid isomers and their conjugated derivatives inhibit growth of human cancer cell lines. AB - Conjugated linoleic acid (CLA), mainly c9,t11- and t10,c12-isomers, and polyunsaturated n-3 fatty acids (n-3 PUFA) have been shown to reduce tumor growth. This study compared, on a set of human tumor cells (breast, lung, colon, prostate and melanoma), the antiproliferative effects of: i) trans monounsaturated fatty acids (MUFA) vs. cis MUFA and MUFA vs. PUFA, ii) individual isomers of CLA vs. linoleic acid, iii) CLA-conjugated derivatives vs. their non conjugated homologues and vs. CLA isomers. Tumor cells were exposed to medium containing individual FA (100 microM) for 48 h and their proliferation was determined by measuring the cellular DNA content (fluorescent Hoechst 33342 dye). The antiproliferative effects of FA varied with the type of cells and were mainly dependent on the degree of unsaturation and on the position and configuration of their double bonds. One isomer of CLA (t9,t11-18:2) and CLA-conjugated derivatives exhibited the strongest growth-inhibitory effect against cancer cells. These results suggest that ruminant products contain active compounds against human tumor cell proliferation. PMID- 16309182 TI - Quantitative structure-cytotoxicity relationship analysis of betulinic acid and its derivatives by semi-empirical molecular-orbital method. AB - A semi-empirical molecular-orbital method (CAChe) demonstrates that the cytotoxicity of betulinic acid derivatives can be predicted by several physical parameters (such as heat of formation, hydrophobicity (log P), watersolubility, ionization potential, electron affinity, dipole moment), but not by molecular size (maximum length and width). The present study demonstrates how this method can be applied to estimate the cytotoxic activity of structurally-related compounds. PMID- 16309183 TI - Clinical significance of double staining of MIB-1 and AgNORs in primary breast carcinoma. AB - BACKGROUND: Argyrophilic nucleolar organizer regions (AgNORs) and MIB-1 as proliferating activities have been applied separately to assess the malignant potential of cancer cells. We conducted staining of AgNORs and MIB-1 in 42 surgically-resected invasive breast carcinomas. MATERIALS AND METHODS: Paraffin embedded sections were used for double staining and the mean AgNOR counts in 100 MIB-1-positive and -negative cells were calculated. RESULTS: The mean AgNOR count in MIB-1-positive cells was significantly higher than in MIB-1-negative ones. AgNOR counts in MIB-1-positive tumors were significantly higher in tumors > or =2 cm and those with positive nodes. Multivariate analysis identified the AgNOR count in MIB-1-positive tumors as the only independent factor related to node metastasis. Survival of patients with lower counts of AgNORs in MIB-1-positive tumors was significantly better compared to those with higher counts. CONCLUSION: Double staining of MIB-1 and AgNORs is useful for predicting lymph node metastasis and prognosis of patients with breast carcinoma. PMID- 16309184 TI - Alteration in platelet function in patients with early breast cancer. AB - The aim of this study was to examine our hypothesis that platelets of patients with breast cancer were functionally altered compared to healthy controls. The results have shown that the platelets from women with early breast cancer released significantly more vascular endothelial growth factor (VEGF) when stimulated with thrombin, tissue factor, clotting, or over a period of time. Similarly, release of thrombospondin (TSP) with thrombin and tissue factor was higher, but failed to reach a significant level. Thus, the observed differences in platelet response support our hypothesis, but warrant further work to determine the reason underlying the observed difference and potential clinical relevance of our findings. PMID- 16309185 TI - Introduction of a survivin gene-specific small inhibitory RNA inhibits growth of pancreatic cancer cells. AB - BACKGROUND: The anti-apoptotic molecule survivin is expressed in human cancers of various origins. Since this molecule possesses multiple functions, including apoptosis inhibition, cell cycle promotion and enhancement of Fas ligand expression, survivin has attracted growing attention as a target in cancer treatment. A survivin-specific small inhibitory RNA (siRNA) was introduced into pancreatic cancer cells to investigate its effect on cancer cell growth. MATERIALS AND METHODS: Survivin mRNA and protein expression were examined by RT PCR and Western blotting, respectively. DNA histogram analysis was performed using a flow cytometer. RESULTS: The introduction of survivin-specific siRNA reduced survivin mRNA and protein expression in PANC-1 cells by over 90% and to an undetected amount, respectively, and induced growth inhibition. The siRNA transfectants showed pronounced morphological changes including enlargement of cells and multinucleation. siRNA transfectants did not show cell cycle arrest, but underwent apoptosis. CONCLUSION: Our data suggest that the use of survivin specific siRNA deserves further investigation as a novel approach to cancer therapy. PMID- 16309186 TI - Antitumor effects of synthetic VEGF-receptor binding antagonist, VGA1155. AB - Vascular endothelial growth factor (VEGF) plays key roles in tumor angiogenesis. Therefore, VEGF and its receptors are considered to be primary targets for antiangiogenic strategy during cancer chemotherapy. Our previous study reported that VGA1155, a low-molecular-weight inhibitor of the binding of VEGF, inhibited VEGF binding to KDR/Flk-1 receptor-overexpressing cells. In the present study, the antitumor effects and antimetastatic effect of VGA1155 were examined in vivo. VGA1155 suppressed the growth of human lung, breast, colon and epidermoid cancers (LC-6, HT29, MX-1, Col-1 and A431) in the nude mouse xenograft model, and pulmonary metastasis of melanoma in the spontaneous metastasis model. These results suggest that VGA1155 has antitumor effects in vivo through the inhibition of VEGF binding to its receptors. PMID- 16309187 TI - Unfiltered xenon light is useful for photodynamic therapy with acridine orange. AB - We have clarified that photodynamic therapy (PDT) with acridine orange (AO) exerts a rapid and strong cytocidal effect on mouse osteosarcomas, both in vitro and in vivo, and have sought to apply this therapy to patients with musculoskeletal sarcomas, in order to reduce the surgical margin and obtain better limb function after tumor resection in limb salvage surgery. Some clinical studies have reported that the local recurrence rate after limb salvage surgery in patients receiving PDT therapy was less than 10% and that the limb functions recovered to nearly normal in these patients. For these basic and clinical studies, we used a blue light beam filtered from a xenon lamp for the AO excitation, because of its maximal absorption. However, the relationship between the cytocidal effect of PDT and the wavelength or illuminance (lux) of the excitation light in AO-PDT is unknown. Therefore, we investigated the cytocidal effects of AO-PDT on mouse osteosarcoma cells using lights of various illuminances and wavelengths from a xenon lamp. Our results revealed that, while the blue and green filtered lights exerted cytocidal effects depending on their illuminance, orange light exerted no such effect. Blue light showed the strongest cytocidal effect under constant illuminance. However, unfiltered light with 10 times the illuminance of blue light yielded a much stronger cytocidal effect, which was deduced not to be due to DNA injury by ultra-violet light or heat generation by ultra-red light, since a xenon lamp emits little of either light. Based on these results, we conclude that, for effective AO-PDT in clinical practice, strong unfiltered light from a xenon lamp is more effective and feasible than weak filtered blue light. PMID- 16309188 TI - Reduction of cisplatin dosage by ZD 1839. AB - Cisplatin (CDDP) is the main chemotherapeutic drug in the treatment of recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN), but its nephrotoxicity often limits the treatment. ZD 1839 is an orally-applicable, selective EGFR tyrosine kinase inhibitor. This investigation explored whether the cisplatin dose can be reduced by the additional application of ZD 1839. Four different SCCHN cell lines were treated with descending doses of CDDP alone or in combination with ZD 1839. Proliferation was measured by the MTT assay; tumor cell toxicity was measured by using the lactate dehydrogenase approach. ZD 1839 augments CDDP-dependent antiproliferative effects. By adding ZD 1839 to the treatment regimen, the CDDP dose could be reduced by up to 25% of the CDDP IC50 dose without compromising the antiproliferative effect. Adding ZD 1839 to CDDP may therefore enable CDDP treatment at a lower dose without compromising antiproliferative effects. PMID- 16309189 TI - Kinetic studies of the radical-scavenging activity of ebselen, a seleno-organic compound. AB - Ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) is now under active investigation as a neuroprotective and anticancer agent. In the present study, the induction period method was used to investigate the antioxidant activity of ebselen in the radical polymerization of methyl methacrylate (MMA) at 70 degrees C. The reaction of ebselen with growing MMA radicals (lipid radicals) showed a k(inh) of 4 x 10(4) M(-1)s(-1). This value was similar to that for mercaptomethylimidazole (MMI, a thiol) and 10-fold greater than that for butylated hydroxyanisole (BHA). The ratio of the rate of chain inhibition to that of chain propagation (CI/CP) for ebselen, MMI and BHA was 0.1, 0.01 and 0.001, respectively, whereas the stoichiometric factor (n, the number of free radicals trapped by one mole of antioxidant moiety) for the corresponding compounds was 0.02, 0.2 and 2, respectively. Ebselen preferentially affected CP rather than CI, indicating that it was an effective scavenger (suppressor) of growing MMA radicals. These results suggest that ebselen is a potent suppressor of polyunsaturated fatty acid (PUFA) radicals, which are harmful radicals in biological systems. PMID- 16309190 TI - Cytogenetic investigations in four canine lymphomas. AB - Four cases of canine lymphoma are presented, including histological examination and cytogenetic investigation. The first case showed a derivative chromosome 13, the second case showed a clonal trisomy 8 and the third case showed a complex karyotype with a clonal trisomy 13 and additional clonal trisomies of the chromosomes 20, 30 and 37, as well as a non-clonal tetrasomy 9. Case four showed a single trisomy 2. Comparing these results with human hematopoietic malignancies, there are notable similarities between both species. PMID- 16309191 TI - Nuclear matrix protein expression in prostate cancer: possible prognostic and diagnostic applications. AB - Different lines of evidence suggest that the nuclear matrix (NM), the protein scaffold of the nucleus, represents a functional unit playing a pivotal role in the spatial and temporal coordination of the events of gene activation. Any change in the gene expression pattern, which occurs during carcinogenesis, may partially depend on an impairment of the regulatory functions of the NM. Therefore, increasing interest has been addressed to the study of NM modifications associated with malignant transformations and to potential clinical applications. Here, recent results on the NM changes in prostate cancer are discussed. Tumor cells are characterized by a more complex NM protein pattern compared to normal tissue: the development of poorly-differentiated tumors is characterized by the expression of proteins that are not present in hyperplastic tissues or in more differentiated tumors. In addition, a few newly-expressed proteins are significantly correlated with the risk of biochemical progression. The potential application of these proteins at the diagnostic and prognostic levels calls for further studies. PMID- 16309192 TI - Evaluation of tumor malignancy in esophageal squamous cell carcinoma using different characteristic factors. AB - BACKGROUND: We have been investigating various molecules correlated with the malignancy of esophageal squamous cell carcinoma and, in the present study, we examined the correlation of four of them (KAI1, FAK, EphA2, Ki-67 labeling index) with the prognosis of affected patients. Furthermore, the use of biopsy samples was studied to evaluate whether the grade of tumor malignancy can be determined before treatment in a clinical setting. MATERIALS AND METHODS: Tissue specimens that had been surgically removed from 91 patients with thoracic esophageal cancer and 247 biopsy samples were examined. The malignancy index (MI) was defined in terms of the KAI1, FAK and EphA2 scores and the Ki-67 labeling index, and the reliability and utility of the correlation between MI and prognosis was evaluated. RESULTS: The mean 5-year survival rate of patients with MI=0 was 100%, while that of patients with MI=1, 2 and 3 was 70%, 48% and 10%, respectively. Patients with MI=4 all died, with the exception of one who has been observed for 3 years. The rate of concordance between the biopsy samples and surgical specimens was 79.4% for KAI1, 88.2% for FAK and 73.5% for EphA2, and the rates of concordance for 1, 2, 3, 4, 5, 6, 7 and 8 biopsy samples were 66.7%, 64.1%, 74.5%, 90.7%, 91.7%, 83.3%, 100% and 100%, respectively. CONCLUSION: It may be feasible to evaluate the malignancy of tumor cells and to predict patient outcome by using multiple marker molecules. It is anticipated that such data will accelerate the development of "tailor-made" therapy. PMID- 16309193 TI - Silencing and CpG island methylation of GSTP1 is rare in ordinary gastric carcinomas but common in Epstein-Barr virus-associated gastric carcinomas. AB - BACKGROUND: The GSTPI gene encodes for glutathione S-transferase pi (GST-pi), which protects cells from cytotoxic agents. The carcinogenic role of this enzyme is at issue because functional polymorphisms have been shown to be a risk factor of human cancer. Moreover, GST-pi protein loss has frequently been reported in various human cancers. MATERIALS AND METHODS: The expression of GST-pi and the methylation status of the promoter area of GST-pi were investigated in gastric carcinomas. Eleven human SNUgastric cancer cell lines, PC-3 prostate cancer cell lines, various cancer tissues and normal gastric mucosa tissues were analyzed by immunohistochemistry, in situ hybridization, Western blot and methylation specific PCR. RESULTS: Only 22 (2.0%o) out of 1081 cases showed loss of GST-pi expression. Interestingly, 16 out of 22 GST-pi-negative cases were Epstein-Barr virus (EBV)-associated gastric carcinomas. The loss of expression of GST-pi among EBV-associated gastric carcinomas was found to be 27.1% (16/59), but to be 0.6% (6/1022) in EBV-negative gastric carcinomas (p < 0.001). Eight out of 16 cases with loss of GST-pi expression showed CpG island methylation in the GSTP1 promoter region, while none of the normal gastric mucosa or EBV-negative gastric carcinomas showed methylation (p < 0.001). CONCLUSION: These findings demonstrate that the loss of GST-pi expression is clustered in a subset of gastric carcinomas with EBV incorporation, and that the methylation of the promoter of the GSTP1 gene is correlated with this loss of GST-pi expression. Our results suggest that GST-pi abrogation by CpG island hypermethylation may account for EBV-associated gastric carcinoma. PMID- 16309194 TI - The effect of o6-methylguanine-DNA methyltransferase (MGMT) and mismatch repair gene (hMLH1) status on the sensitivity to alkylating agent 1-(4-amino-2-methyl-5 pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea(ACNU) in gallbladder carcinoma cells. AB - The aim of this study was to correlate O6-methylguanine-DNA methyltransferase (MGMT) and mismatch repair gene (hMLH1) expression in gallbladder carcinoma cells with drug sensitivity to the bifunctional alkylating agent 1-(4-amino-2-methyl-5 pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea (ACNU). Using 5 gallbladder carcinoma cell lines and 1 colon carcinoma cell line (SW48), the MGMT and hMLH1 status was assessed both by RT-PCR and Western blot analyses. Sensitivity to ACNU was estimated by the MTT assay. MGMT+/hMLH1+ status was revealed in 2 gallbladder carcinoma cells, MGMT-/hMLH1+ in another 3 gallbladder carcinomas and MGMT-/hMLH1 in SW48. MGMT-/hMLH1+ and MGMT-/hMLH1- cells were more sensitive to ACNU compared with MGMT+/hMLH1+ cells. These results indicate that MGMT, but not hMLH1, expression is an important determinant for drug sensitivity to ACNU in gallbladder carcinoma cells. The drug effect of ACNU, which depends on the MGMT status, was verified using xenograft tumors grown in nude mice. Furthermore, the apoptotic index of MGMT- GB-dl xenografts was significantly increased by ACNU treatment, compared with that of MGMT+ KMG-C. In conclusion, the sensitivity to ACNU was not associated with hMLH1 status, but was found to depend only on the MGMT status. ACNU might be a useful chemotherapeutic agent for MGMT- gallbladder carcinoma, which is associated with poor patient outcome. PMID- 16309195 TI - Induction of cytotoxicity and apoptosis and inhibition of cyclooxygenase-2 gene expression, by curcumin and its analog, alpha-diisoeugenol. AB - Cytotoxici and alpha-diisoeugenol were investigated. The cytotoxicity of curcumin and a-diisoeugenol against human promyelocytic leukemia cells (HL-60 cells) and human submandibular cancer cells (HSG cells) was similar (CC50 1-3 microM). However, curcumin induced much more apoptosis, particularly in HL-60 cells compared with HSG cells, as revealed by measurement of the sub-G1/G0 DNA fraction in flow cytometric histograms. Treatment with 15 microM curcumin increased the number of cells with a sub-G1/G0 DNA fraction from control levels of <5% to 55% in HL-60 cells and 30% in HSG cells. Flow cytometry, after staining with annexin V-FITC/PI (the exposure of phosphatidylserine (PS) on the surface of apoptotic cells), showed a dose-dependent induction of early apoptosis by curcumin, which reached about 65% in HL-60 cells and about 20% in HSG cells after treatment with 10 microM curcumin. In contrast, alpha-diisoeugenol failed to induce apoptosis in either cell type. For both cell types, the proportion of late apoptotic/necrotic cells increased rapidly at concentrations of curcumin and a-diisoeugenol greater than 10 microM. The generation of intracellular reactive oxygen species (ROS) in curcumin-treated HL-60 cells was greater than that in HSG cells, as judged by CDFH-DA staining. In both cell types, ROS generation by a-diisoeugenol was at control levels. ROS generation by curcumin was suppressed by antioxidants such as N-acetyl-L-cysteine (NAC) and glutathione (GSH) and by scavengers of hydroxy radicals such as mannitol, but, conversely, was promoted by prooxidants such as the transition metal ions Cu(II) and Zn(II). ROS generation may play a part in the exposure of PS. Curcumin, but not a-diisoeugenol, at 10 microM inhibited LPS (lipopolysaccharide)-induced COX-2 gene expression in RAW 264.7 cells. Semiempirical PM 3 calculations suggested that this activity of curcumin, in which it behaves as a non-steroidal anti-inflammatory drug (NSAID)-like compound, is dependent on its phenolic function, which is more pronounced than that of alpha-diisoeugenol. Taken together, our results suggest that the bioactivity of curcumin is a result of its ability to act as both a prooxidant and an antioxidant. PMID- 16309196 TI - Effect of anticancer agents on codeinone-induced apoptosis in human cancer cell lines. AB - The possible apoptosis-inducing activity of codeinone, an oxidative metabolite of codeine, without or with anticancer drugs, was investigated. Codeinone induced internucleosomal DNA fragmentation in human promyelocytic leukemia cells (HL-60), but not in human squamous cell carcinoma cells (HSC-2). Codeinone dose dependently activated caspase-3 in both of these cells, but to a much lesser extent than that attained by actinomycin D. This property of codeinone was similar to what we have found previously in alpha,beta-unsaturated ketones. Codeinone did not activate caspase-8 or caspase-9 in these cells. The cytotoxic activity of codeinone against HSC-2 cells was inhibited by N-acetyl-L-cysteine, but somewhat additively stimulated by sodium ascorbate, epigallocatechin gallate, hydrogen peroxide, sodium fluoride, 5-fluorouridine, cisplatin, doxorubicin and methotrexate. These data suggest that codeinone has possible antitumor potential, in addition to its action as a narcotic analgesic, even though it induces incomplete apoptosis-associated characteristics. PMID- 16309197 TI - Glycyrrhetinic acid and related compounds induce G1 arrest and apoptosis in human hepatocellular carcinoma HepG2. AB - Glycyrrhetinic acid (GA) and its related compounds are known to have anti inflammatory activity and also to inhibit liver carcinogenesis and tumor growth. GA and related compounds inhibited cell proliferation of the human hepatoma cell line, HepG2. Among five compounds tested, ursolic acid and 18beta-olean-12-ene 3beta, 23, 28-triol (18beta-erythrotriol) were comparatively effective, where the 50% inhibitory dose was 20 microM and 25 microM, respectively. Flow-cytometric analysis showed that GA and the related compounds arrested the cell cycle in the G1-phase; in addition, GA-related compounds induced apoptosis at high dose. Western blot analysis indicated that the induction of apoptosis by GA and ursolic acid was accompanied with an activation of caspase-8 and a reduction in the anti apoptotic proteins, Bcl-2 and Bcl-xL, although the pro-apoptotic proteins, Bax and Bak, remained unaffected. These results suggest that GA and its related compounds may be potent agents in liver cancer treatment. PMID- 16309198 TI - Under-representation of bladder transitional cell tumour 9q, 11p and 14q LOH in urine and impact on molecular diagnosis. AB - BACKGROUND: To investigate whether the recently reported evidence of differences in the overall loss of heterozygosity (LOH) frequency between urine and tumour tissue in patients with transitional cell tumours (TCT) of the urinary bladder involved specific chromosomal sites, and their impact in diagnosis. MATERIALS AND METHODS: Blood, tissue and urine specimens were obtained from 55 patients and 25 controls. Sixteen microsatellites were PCR-amplified and blindly analyzed for LOH through a laser-based capillary electrophoresis system. RESULTS: Significant frequence differences between tumour tissue and urine sediment LOH were found in 9q and 11p in non-invasive disease and 14q in invasive disease. There was no significant difference for all the other chromosomal arms analyzed. CONCLUSION: The contribution in the urine sediment of cells belonging to tumours of the same histological classification differs according to the specific genetic alterations these cells carry. Furthermore, the location regarding these differences could indicate regions involved in tumour exfoliation or apoptosis. PMID- 16309199 TI - Tumor-specific cytotoxicity and apoptosis-inducing activity of berberines. AB - Berberine iodide (IK-1) and acetoneberberine (IK-2) showed higher cytotoxicity against five human oral squamous cell carcinoma (HSC-2, HSC-3, HSC-4, NA, CA9-22) and one human promyelocytic leukemia (HL-60) cell lines, than against normal human oral tissue-derived cells (gingival fibroblast HGF, pulp cell HPC, periodontal ligament fibroblast HPLF), producing a tumor specificity index of 4.0 and 3.6, respectively. IK-1 was more potent than IK-2 in inducing the production of apoptotic cells, internucleosomal DNA fragmentation, the activation of caspases-3, -8 and -9, and the increased expression of proapoptotic BAD protein, with a corresponding decrease in the expression of anti-apoptotic Bcl-2 protein in HL-60 cells. These compounds did not induce internucleosomal DNA fragmentation (only producing larger DNA fragment), nor increased the Bad protein expression in HSC-2 cells. The present study demonstrated the tumor-specific cytotoxicity and apoptosis-inducing activity of berberines, suggesting their possible antitumor potentiaL PMID- 16309200 TI - Aneuploidy of chromosome 8 and C-MYC amplification in individuals from northern Brazil with gastric adenocarcinoma. AB - BACKGROUND: Gastric cancer is the third most frequent type of neoplasia. In northern Brazil, the State of Para has a high incidence of this type of neoplasia. Limited data are available so far on the genetic events involved in this disease. MATERIALS AND METHODS: Dual-color fluorescence in situ hybridization (FISH) for the C-MYC gene and chromosome 8 centromere was performed in 11 gastric adenocarcinomas. RESULTS: All cases showed aneuploidy of chromosome 8 and C-MYC amplification, in both the diffuse and the intestinal histopathological types of Lauren. No correlation was found between polysomy 8 and the histopathological characteristics of the tumors. C-MYC amplification, like homogeneously-stained regions (HSRs) and double minutes (DMs), was observed only in the intestinal-type. Translocation of C-MYC was observed only in the diffuse-type. CONCLUSION: Chromosome 8 can be used as a marker in the diagnosis of gastric adenocarcinoma. The C-MYC oncogene requires further studies in order to verify if it is, when amplified, an etiological cause of transformation or a consequence of the proliferation process. PMID- 16309201 TI - Generation of an autologous cell system for immunotherapy of squamous cell carcinoma of the head and neck. AB - BACKGROUND: To date, there is no tumor antigen known to be sufficiently specific for diagnosis, therapy monitoring and immunotherapy of squamous cell carcinoma of the head and neck (SCCHN). The aim of our study was to generate an autologous immune response against SCCHN in vitro for further characterization of SCCHN specific tumor markers and adoptive immunotherapy. MATERIALS AND METHODS: As sources for tumor antigens (Ags) for the restimulation of autologous immune cells, cell lines from solid SCCHN were established and characterized. Forty-five percent of 40 tumors of different SCCHN specimens were maintained for more than 20 cell generations in culture. RESULTS: One primary cell line, SCCHN-GHD, newly established from a hypopharynx carcinoma, was further characterized as a telomerase-positive, immortalized cell line with epithelial cell characteristics. It was found to be tumorigenic in SCID mice. CONCLUSION: This new SCCHN-GHD cell line is competent as a target for lysis by autologous immune cells and for the restimulation of autologous tumor-specific immune cells. Subsequent characterization of tumor antigens will be performed. PMID- 16309202 TI - Gamma-irradiation induced apoptosis in peritoneal macrophages by oxidative stress. Implications of antioxidants in caspase mitochondrial pathway. AB - The in vivo and in vitro development of apoptosis induced by gamma-irradiation was studied in mouse peritoneal macrophages. The apoptosis index was measured by fluorescence microscopy and DNA electrophoresis. In vivo apoptosis was greatest eight days after 8 Gy total body gamma-irradiation. A DNA ladder electrophoretic pattern was only observed in the gamma-irradiated group. The participation of reactive oxygen species in apoptosis induction was investigated by pretreating mice with the antioxidants superoxide dismutase, catalase, vitamin E or lipopolysaccharide before gamma-irradiation. Measurement of serum lipoperoxides showed oxidative stress in the gamma-irradiated mice and the protection given by the antioxidants. These results were confirmed using in vitro cultures of peritoneal macrophages: gamma-irradiated groups and antioxidant-pretreated gamma irradiation groups showed results similar to those observed with in vivo irradiation. A loss of mitochondrial membrane potential (delta psi(m)) was also observed by microscopy in the gamma-irradiated cell cultures. Experiments with caspase inhibitors confirmed the participation of caspase 3 and caspase 9. PMID- 16309203 TI - Prognostic value of matrix metalloproteinase-2 (MMP-2) expression in endometrial endometrioid adenocarcinoma. AB - Matrix metalloproteinase-2 (MMP-2), a member of the zinc-dependent metalloproteinase gene family, plays an important role in cancer invasion and metastasis. The current study aimed to evaluate whether the expression of MMP-2 is associated with survival in patients with endometrial endometrioid adenocarcinoma. The MMP-2 immunoreactive protein was evaluated from endometrioid adenocarcinoma of the endometrium in 112 patients treated at Oulu University Hospital, Finland. The median follow-up time was 88 months. The expression of MMP 2 was studied immunohistochemically in paraffin-embedded tissue samples from the primary tumours by using a specific monoclonal antibody to MMP-2. The MMP-2 protein was found in 80% of the primary tumours, including all histological grades. All grade 3 tumors were MMP-2-positive. At the end of the study period, 21 of the 22 (95%) patients presenting MMP-2-negative immunostaining were alive, whereas the corresponding figure for those with MMP-2-positive tumours was 78 out of 90 (87%). These data suggest that MMP-2 immunostaining negativity might be linked with a favourable prognosis in endometrial endometrioid adenocarcinoma. PMID- 16309204 TI - Development of sex cord stromal tumors after heterotopic transplantation of cryopreserved ovarian tissue in rats. AB - Chemoradiotherapy has substantially improved life expectancy in young women with cancer, but these treatments often cause infertility. One method of preserving fertility is to cryopreserve ovarian tissue, with subsequent autotransplantation of the tissue after successful anticancer therapy. This study examined the long term effect of heterotopic transplantation of cryopreserved ovarian tissue on the histology. Ovarian tissue from rats was cryopreserved using a slow-freezing protocol. After thawing, the tissue pieces were transplanted under the splenic capsule in 14 rats of the same inbred strain and remained there for 210 or 300 days. Sex cord stromal tumors, consisting mainly of granulosa cells, were found in all of the rats. Although the hormonal situation in rats cannot be directly compared to that in humans, the development of sex cord stromal tumors in this animal model may be worth considering when cryopreserved ovarian tissue is transplanted heterotopically in fertility-preserving programs for cancer patients. PMID- 16309205 TI - Different roles for K+ channels in cisplatin-resistant cell lines argue against a critical role for these channels in cisplatin resistance. AB - Cisplatin resistance has been associated with altered K+ fluxes. Here, we focused our investigations on the detection of K+ channels in a series of cisplatin resistant (CP-r) cells with increasing resistance and on the functional relationship of these K+ channels to resistance. Microarray analysis and confocal microscopy indicated that there was overexpression of the ether-a-gogo gene (HERG) and the inwardly rectifying potassium channel gene (TWIK) in a human epidermal KB and human liver BEL-7404 carcinoma cell line series selected for cisplatin resistance. With increased resistance, the plasma membrane potential, but not the mitochondrial membrane potential, also increases in these two series. For these reasons, we conducted cell proliferation studies in the presence of either antibodies directed against the detected K+ channels, omeprazole (a H+ pump inhibitor) or a specific inhibitor of the HERG channel (WAY-123398-A-5). The antibodies and omeprazole influenced cell growth only very slightly. The specific K+ channel blocker did not alter cisplatin resistance. We also observed that manipulation of K+ fluxes with antibodies and the H+ pump with omeprazole resulted in opposite effects on cisplatin resistance in these two cell lines. We conclude that K+ and H+ homeostasis are not critical factors in cisplatin resistance since they affect cisplatin resistance differently in KB and BEL-7404 cells. PMID- 16309206 TI - Morphological and histochemical evidence of the protective effect of procainamide hydrochloride on tissue damage induced by repeated administration of low doses of cisplatin. AB - BACKGROUND: The class 1 antiarrhythmic drug procainamide hydrochloride might protect against acute cisplatin-induced nephrotoxicity and hepatotoxicity in mice and rats. In this report, the protective activity of procainamide hydrochloride against renal and hepatic tissue damage induced by repeated administration of low doses of cisplatin was analyzed morphologically and histochemically. MATERIALS AND METHODS: Light microscopy observations were performed on liver, renal and heart samples obtained from female Wistar rats treated twice a week for 10 weeks with 1 mg/kg cisplatin (cumulative dose: 20 mg/kg), with or without 100 mg/kg procainamide hydrochloride (cumulative dose: 2 g). Samples were then submitted to histochemical stainings [i.e. H & E, periodic acid Schiff (PAS) and Sudan Black]. RESULTS: Light microscopy analysis revealed that the coadministration of cisplatin and procainamide hydrochloride significantly reduced tissue alterations both in the kidneys and liver, while in the heart, neither cisplatin nor the combination of cisplatin and procainamide hydrochloride caused any evident tissue damage. CONCLUSION: The morphological and histochemical data confirm that procainamide hydrochloride is able to protect not only from acute cisplatin induced toxicities, but also from tissue alterations induced in the liver and kidneys by the administration of repeated low doses of cisplatin. PMID- 16309207 TI - Influence of matrix metalloproteinase 9 (MMP-9) on the metastatic behavior of oropharyngeal cancer. AB - BACKGROUND: The role of the single matrix metalloproteinases (MMPs) in the metastatic process of squamous cell carcinomas (SCC) is still obscure. MATERIALS AND METHODS: The MMP-9 expression was described immunohistochemically in 105 patients (40-79 years of age, mean: 57.84 years; 84 male, 21 female) suffering from orophatyngeal cancer (22x TI, 31x T2, 24x T3, 28x T4) with different neck stages (41x N0, 6x N1, 54x N2, 4x N3 neck). RESULTS: A significant correlation between MMP-9 expression and T stage (p < 0.05), N stage (r = 0.55, p < 0.01) and UICC stage (r = 0.55, p < 0.01) was revealed. Most remarkable was the high MMP-9 expression with simultaneously high UICC stages. CONCLUSION: The results give further indication that MMP-9 plays a role in the metastatic behavior of oropharyngeal SCC. It will be a project for the near future to create a standardized evaluation score of immuno-histological stainings to allow valid comparison of the results and published data. PMID- 16309208 TI - Characterization of the focal adhesion complex in human non-small cell lung cancer cell lines. AB - BACKGROUND: The important metastatic potential of lung cancers is directly correlated with cell adhesion. Cell-extracellular matrix interactions occur in specialized structures termed focal adhesion (FA) complexes. Our aims were to investigate: (i) the expression of the major FA components in three lung cancer cell lines (non metastatic: A549, or metastatic: Calu-1 and H460), (ii) the modifications of the FA complex occurring when apoptosis was induced by Vinorelbine in the A549 cells. MATERIALS AND METHODS: The FA complex was characterized by flow cytometry, immunocytochemical staining and Western blot. RESULTS: The expressions of alpha3, betsaP, paxillin, p-paxillin and Grb2 varied depending on the histological type of the tumor. In apoptotic cells, the expressions of the PYK2, p-p38, PI3K and Grb2 adhesion proteins were increased. CONCLUSION: Our data suggest that these adhesion proteins may be implicated in the transduction of death signals. PMID- 16309209 TI - The relevance of adhesion molecules in the classification of squamous cell carcinoma of the head and neck. AB - BACKGROUND: Clinical studies have demonstrated increased serum levels of E selectin, P-selectin and VCAM-1 in patients with squamous cell carcinomas, which correlate with expression in the primary tumour. For this reason, selectin expression may also support the diagnosis and be of prognostic value in squamous cell carcinoma of the head and neck (SCCHN). MATERIALS AND METHODS: Using immunohistochemistry, the expressions of E- and P-selectin and VCAM-1 in SCCHN were characterised and analysed. RESULTS: The expressions of E-selectin and VCAM 1 were increased. The magnitude of expression was unrelated to the location or size of the primary tumour. With increasing de-differentiation, the expression of E-selectin increased, whereas the number of cells expressing VCAM-1 was highly variable. In contrast, E-selectin and VCAM-1 expressions decreased with progressive metastatic disease. CONCLUSIONS: These data demonstrate that the expression patterns of E-selectin and VCAM-1 may depend on differentiation and lymphogenic metastatic disease in SCCHN, and suggest that these molecules may have a dual role, depending on the stage of tumour development. These findings support the hypothesis that expression of E-selectin and VCAM-1 may be initiated by the release of tumour necrosis factor-alpha (TNF-alpha) at an early stage of tumour development. In contrast, the decrease in selectin expression, observed as the metastatic disease progresses, may serve as a protective mechanism to prevent an influx of effector cells. PMID- 16309210 TI - Berberine decreased N-acetylation of 2-aminofluorene through inhibition of N acetyltransferase gene expression in human leukemia HL-60 cells. AB - Previous studies have demonstrated that berberine decreases N-acetyltransferase (NAT) activity in human leukemia HL-60 cells, however, NAT gene expression has not been investigated. In this study, berberine was selected for testing the inhibition of N-acetylation of 2-aminofluorene (AF) and NAT gene expression in human HL-60 cells. The N-acetylation of AF was determined and quantitated by high performance liquid chromatography (HPLC). The data showed that a 24-hour berberine treatment decreased the amount of N-acetylation of AF in HL-60 cells. The NAT enzymes were stained and examined by Western blotting and flow cytometry. The results indicated that berberine decreased the levels of NAT protein in HL-60 cells. The expression of NAT gene (mRNAT NAT1) was determined by polymerase chain reaction (PCR), and it was found that berberine had inhibited the expression of mRNA NAT1 in human HL-60 cells. PMID- 16309211 TI - Inhibition of LPS-stimulated NO production in mouse macrophage-like cells by azulenequinones. AB - Azulenequinone derivatives have been reported to display a broad spectrum of biological activities, but study at the cellular level has been limited. The effect of twenty-seven azulenequinone derivatives on nitric oxide (NO) production by mouse macrophage-like cells Raw 264.7 was investigated in this study. All of these compounds failed to stimulate the Raw 264.7 cells to produce detectable amounts of NO, but did inhibit NO production by lipopolysaccharide (LPS) activated Raw 264. 7 cells to varying extents. Compounds [7, 8, 9, 13, 16, 25, 27], which showed lesser cytotoxic activity (CC50 = 425, 381, 482, 179, 119, 235, 225 microM, respectively), inhibited NO production to the greatest extent [selectivity index (SI) = 15.4, 26.2, 3.9, 21.6, 3.1, 6.0, 8.4, respectively]. Western blot and RT-PCR analyses demonstrated that the most active derivatives, 3 morpholino-1, 5-azulenequinone [8] and 3,7-dibromo-1, 5-azulenequinone [13], significantly reduced both the intracellular concentration of iNOS protein and the expression of iNOS mRNA. ESR spectroscopy showed that compounds [8, 13] weakly scavenged NO produced by NOC-7, possibly via their general reducing activity. These data suggest that the inhibitory effect of NO production by compounds [8, 13] might be generated mostly via the inhibition of iNOS expression, rather than the radical-mediated mechanism. PMID- 16309212 TI - Paclitaxel decreases the accumulation of gemcitabine and its metabolites in human leukemia cells and primary cell cultures. AB - BACKGROUND: We identified a drug-drug interaction between gemcitabine and paclitaxel in a clinical pharmacokinetic study. The purpose of the present study was to determine whether paclitaxel affected the uptake and accumulation of the parent drug gemcitabine and the formation of its metabolites after treatment of cells with gemcitabine in vitro. MATERIALS AND METHODS: The human leukemia cell line CEM was treated with 15 micoM 3H-gemcitabine, with and without paclitaxel, and the accumulation of radiolabeled gemcitabine was assessed up to one minute and one hour. Peripheral blood mononuclear cells (PMN) and hepatocytes were treated with gemcitabine, with or without paclitaxel, for specified amounts of time at three concentrations of gemcitabine, and the concentrations of gemcitabine and its metabolites were measured by liquid chromatography. RESULTS: In CEM cells, paclitaxel reduced the uptake and accumulation of gemctabine by 32% and 30%, respectively. In the hepatocytes, the mean concentrations of gemcitabine increased in the cell culture media 100%, 48% and 38% when treated with paclitaxel plus gemcitabine 5, 15 and 30 microM, respectively, compared to gemcitabine alone. The concentrations of the deaminated metabolite dFdU were significantly decreased in the cell culture media. In the PMN, the intracellular accumulation of active triphosphorylated metabolite dFdCTP was lower in cells treated with paclitaxel (up to 83%) compared to the control. CONCLUSION: Paclitaxel substantially reduced the uptake and accumulation of gemcitabine and the formation of its metabolites in vitro at clinically relevant concentrations. PMID- 16309213 TI - The effects of jatrophane derivatives on the reversion of MDR1- and MRP-mediated multidrug resistance in the MDA-MB-231 (HTB-26) cell line. AB - Multidrug resistance (MDR) of cancer cells can be the result of a variety of mechanisms that are not completely understood. One of the most significant among them concerns altered membrane transport in tumor cells, often referred to as typical or classic MDR. This mechanism is related to the overexpression of a variety of proteins, that belong to the super family of ABC transporters. The aim or this study was to look for new effective modulators of MDR1 and multidrug resistance-associated protein (MRP) transporters. Ten diterpenes based on the jatrophane skeleton, including rearranged polycyclic derivatives, were studied on the MDA-MB-231 (HTB-26) human breast cancer cell line. The majority of those compounds were able to strongly enhance the rhodamine 123 accumulation of the human MDR1 gene transfected mouse lymphoma cell line, as previously described. In the present study, the MDR reversal of the same jatrophanes on MDR1- and MRP- mediated resistance of human breast cancer cells is reported. These cells simultaneously express MDR1 and MRP proteins when identified by monoclonal antibodies. However, in a functional assay, where rhodamine 123 accumulation was measured and verapamil was the traditional positive control, only MRP was active, while MDR1 was inactive. Carboxyfluorescein served as a substrate for MRP mediated drug efflux, and indomethacine was the positive control used as an inhibitor of MRP in the flow cytometric experiments. The effectivity of various jatrophanes was different on the carboxyfluorescein efflux inhibition of the human breast cancer cells. These results may have importance in the planning of a new type of combination chemotherapy. PMID- 16309214 TI - Different ways to induce apoptosis by fenretinide and all-trans-retinoic acid in human B lymphoma cells. AB - All-trans-retinoic acid (ATRA) and its synthetic analog fenretinide (4HPR) are potent anticancer drugs. Only a few reports are available about the effects of retinoids on B lymphoma cells. In our study, non-Hodgkin lymphoma cells (HT58) were treated with ATRA and 4HPR. Both agents induced cell death time- and dose dependently. Reactive oxygen species (ROS) production was elevated in 4HPR treated cells, but not in ATRA-treated cells. The depolarization of the mitochondrial membrane occured earlier after ATRA than after 4HPR treament. Z-VAD fmk, the general caspase inhibitor, decreased the DNA fragmentation in ATRA treated cells, but simultaneously increased necrosis. However, z-VAD-fmk did not influence the DNA fragmentation in 4HPR-treated cells. Endonuclease G was released from the mitochondria during 4HPR treatment, which could be an inducer for caspase-independent DNA fragmentation. Our results suggest that natural (ATRA) and synthetic (4HPR) retinoids induce different apoptotic pathways in B lymphoma cells, which is particularly relevant for their potential use in leukemia treatment. PMID- 16309215 TI - Synergistic role of arabinoxylan rice bran (MGN-3/Biobran) in S. cerevisiae induced apoptosis of monolayer breast cancer MCF-7 cells. AB - We have recently demonstrated that breast cancer cell (BCC) lines undergo apoptosis following phagocytosis of S. cerevisiae. Arabinoxylan rice bran extract (MGN-3/Biobran) has been shown to enhance this effect. Since previous data were obtained using cells in suspension, the present study was undertaken to examine monolayer BCC that more closely model cancer cell growth. Monolayers of both breast cancer (MCF-7) and non-tumorgenic breast epithelial (MCF-10A) cells grown on glass coverslips were cultured with heat-killed S. cerevisiae at a ratio of 1:10, respectively. MCF-7 cells phagocytized yeast in a time-dependent manner, 6.9% to 14.3% from 1 h to 4 h, respectively, with a 2-fold increase in the presence of MGN-3. On the other hand, there was virtually no phagocytosis of yeast by MCF-10A cells. Similarly, yeast-induced apoptosis of MCF-7 cells occurred in a time-dependent manner, from 11.5% after 1 h to 21.7% after 4 h, and was enhanced in the presence of MGN-3. These data may have implications in the treatment of breast cancer. PMID- 16309216 TI - Caspase-3 activation and induction of PARP cleavage by cyclic dipeptide cyclo(Phe Pro) in HT-29 cells. AB - BACKGROUND: Cyclo(Phe-Pro) has been shown to inhibit cancer cell growth and induce apoptosis in HT-29 colon cancer cells. MATERIALS AND METHODS: The molecular mechanisms mediating cyclo(Phe-Pro)-induced apoptosis in HT-29 cells were investigated. Cells were treated with 5 mM or 10 mM cyclo(Phe-Pro) for varying times. Immunoblot analysis was used to detect poly(ADP-ribose)polymerase (PARP) cleavage. A fluorescence-based enzymatic assay was used to measure caspase 3 activity. RESULTS: Cyclo(Phe-Pro) (10 mM) induced time-dependent cleavage of PARP, detected as early as 8 hours post treatment. PARP cleavage was blocked by co-administration with the broad-range caspase inhibitor Z-VAD-FMK Cyclo(Phe-Pro) also induced a time-dependent increase (p < 0.01) in caspase-3 activity. This increase in activity was blocked in the presence of the caspase-3 inhibitor Ac DEVD-CHO. CONCLUSION: These results provide evidence that cyclo(Phe-Pro)-induced apoptosis in HT-29 cells is mediated by a caspase cascade. These findings warrant further investigation into the potential antitumour activity of cyclo(Phe-Pro) and its related cyclic dipeptide derivatives. PMID- 16309217 TI - Induction of apoptosis and cell cycle arrest in glioma cells by GL331 (a topoisomerase II inhibitor). AB - BACKGROUND: GL331 is a topoisomerase II inhibitor and, in this study, the effects of GL331 upon rat C6 glioma cells were investigated. MATERIALS AND METHODS: The glioma cells were treated with GL331, then a cytotoxicity assay was done to evaluate the cytotoxic effects of GL331 and flow-activated cell sorter analysis was performed to analyze the cellular apoptosis and cell cycle distribution of the glioma cells. The expressions of p53, p21 and 14-3-3sigma in glioma cells treated with GL331 were investigated by Western blot analysis. RESULTS: GL331 was demonstrated to exert cytotoxic effects upon and induce apoptosis of the C6 glioma cells in a concentration- and time-dependent manner (p < 0.05). GL331 also caused the glioma cells to accumulate in the G2/M-phase of the cell cycle and increased the expressions of p53, p21 and 14-3-3sigma. CONCLUSION: GL331 exerts cytotoxic effects and induces apoptosis in C6 glioma cells, accompanied by cell accumulation in the G2/M-phase and activation of p53, p21 and 14-3-3sigma. PMID- 16309218 TI - The auricular VX2 carcinoma: feasibility of complete tumor resection. AB - BACKGROUND: The aim of the study was to test the feasibility of resection of VX2 auricular squamous cell carcinoma (SCC) in New Zealand White rabbits (NZW), its influence on the regional and distant metastases and the general prognosis. MATERIALS AND METHODS: In 71 NZW rabbits, successful in sano wide margin resection of auricular VX2 SCC was performed 7 days after tumor induction. The animals were sacrificed and examined for metastases after 6 weeks. RESULTS: Local recurrences occured in 7% of the animals. In 28.2% lymph node (LN) and in 15.5% distant metastases developed. Regional metastases presented in 100% of the animals with recurrence and in 22.7% of the RO animals (p < 0.001). Distant metastases presented in 80% of the animals with local recurrence and in 10.6% of the R0 animals (p < 0.001). Of the animals with local control, those with N+ had a significantly higher incidence of distant metastases (46.7%) compared to those with N0 (0%, p < 0.001). Local control and LN involvement were found to be important prognostic factors for early tumor-related death. CONCLUSION: The VX2 auricular model resembles head and neck (HN) SCC in humans morphologically, pathophysiologically and as a complex neoplastic disease. In the experimental setting it behaved similarly to the clinical situation of wait-and-see in N0 HNSCC patients. Tumoral microemboli present a possible mechanism for the development of recurrences. PMID- 16309219 TI - Cell death induced by taxanes in breast cancer cells: cytochrome C is released in resistant but not in sensitive cells. AB - BACKGROUND: The aim of the study was to contribute to our understanding of the mechanisms responsible for the resistance of breast cancer cells to taxanes. MATERIALS AND METHODS: Cell cycle characteristics, DNA fragmentation, p53 and p21(WAF1/CIP1) expression, caspase-3 and caspase-9 activity, cytochrome c release from mitochondria during cell death induction by the taxanes paclitaxel and docetaxel in highly-sensitive MDA-MB-435 and highly-resistant NCI-ADR-RES human breast cancer cells were compared. RESULTS: Approximately 300-fold higher concentrations of the taxanes were required to induce death in resistant NCI-ADR RES cells than in sensitive MDA-MB-435 cells. Cell death induced by the taxanes in both sensitive and resistant cells was preceded by the accumulation of cells in the G2/M-phase. Neither cell type produced any DNA fragmentation (DNA ladder) typical of regular apoptosis. The p53 and the p21(WAF1/CIP1) levels did not change in sensitive or in resistant cells during cell death induction by the taxanes. The activity of the executioner caspase-3 increased significantly (2 to 2.5-fold) and, similarly, the activity of caspase-9 increased significantly (2 to 3.5-fold) in both cell types. However, cytochrome c was found to be released from mitochondria into the cytosol only in the resistant NCI-ADR-RES cells, but not in the sensitive MDA-MB-435 cells. CONCLUSION: The death induced by the taxanes in the studied breast cancer cells can be characterized as an apoptosis-like death, including caspase-3 and caspase-9 activation but not oligonucleosomal DNA fragmentation. However, the mechanisms of death induction by the taxanes in sensitive MDA-MB-435 cells and resistant NCI-ADR-RES cells differ. Cytochrome c is released from the mitochondria in resistant but not in sensitive cells. PMID- 16309220 TI - Medullary thyroid carcinoma: autologous tumor cell lines for dendritic cell vaccination. AB - BACKGROUND: Medullary thyroid carcinoma (MTC) is a calcitonin-producing tumor of the parafollicular C-cells, accounting for 5-10% of all thyroid tumors. To date, the only effective treatment is the early and total surgical removal of all neoplastic tissue. As the prognosis of patients with advanced MTC, unresectable or distant metastases is poor, and chemotherapy or irradiation is of no significant value, alternative strategies have been sought. MATERIALS AND METHODS: A promising treatment approach for human MTC, that has already been introduced at our facility, is based on vaccination with autologous dendritic cells (DCs). Strong evidence that vaccination with autologous tumor lysate-pulsed DCs induces a specific immune response in vivo has been provided. However, the therapeutic success of this approach is sometimes critically limited by the small amount of tumor material available, especially from patients operated at an early tumor stage. Thus, it would be to the best advantage to have sufficient amounts of autologous tumor cells available for DC pulsing. RESULTS: A method to generate viable autologous tumor cell cultures from a variety of MTC tissue samples, even when the sample size is small, has been successfully established. These cell lines maintain their neuroendocrine phenotype. In addition, it can be shown that these cells also display the biological features of neuroendocrine tumor cells at the molecular level. CONCLUSION: The unlimited availability of these MTC cell lines makes it possible to specify cancerogenesis of MTC. In addition, the availability of sufficient amounts of tumor lysate from these cell lines offers the advantage of prolonged immunotherapy. Finally, these cell lines could be elegantly used as read-out system to monitor the in vivo immune response during immunotherapy with DC cell-based vaccination in patients suffering from MTC. PMID- 16309221 TI - Understanding the response of dendritic cells to activation by streptococcal preparation OK-432. AB - BACKGROUND: The streptococcal preparation OK-432 induces maturation of T cells and dendritic cells (DCs). However, the mechanisms by which OK-432 induces DC maturation are not well understood. MATERIALS AND METHODS: The effects of OK-432 and TNF-alpha on peripheral blood mononuclear cells (PBMCs) were compared. Antibody-based approaches were used to detect proteins characteristic of antigen presenting cells and cytokines. Cytotoxicity was evaluated by measuring the release of LDH after incubation of effector and target cells. The TLR-4 levels were measured by real-time quantitative PCR. RESULTS: Changes in cell surface marker levels were detected in both treatment groups but OK-432 had a greater effect on induction of Th-1-type cytokines. Furthermore, TLR-4 mRNA was up regulated in cells from two out of five patients in response to OK-432, but not in response to TNF-alpha. CONCLUSION: OK-432 has a more profound effect on human DCs than TNF-alpha and may act through multiple signaling pathways. PMID- 16309222 TI - Prednisolone reduces TNF-alpha release by PBMCs activated with a trifunctional bispecific antibody but not their anti-tumor activity. AB - BACKGROUND: New adjuvant immunological therapies, that selectively redirect effector cells towards tumors, are currently under development. These strategies include trifunctional bispecific antibodies (trAb) as promising tools for the elimination of disseminated tumor cells and micrometastases. To date, these chimeric molecules have demonstrated their antitumor potential mainly in vitro. Here, trAb-activated peripheral blood mononuclear cells (PBMCs) displayed considerable antitumor activity, accompanied by the release of cytokines, which contributed to the antitumor activity but, on the other hand, may evoke serious limiting side-effects in vivo, demanding therapeutic interventions. MATERIALS AND METHODS: The antitumor activity and cytokine release by trAb-activated PBMCs were studied in co-cultures with multicellular tumor spheroids (MTS), which represent a three-dimensional in vitro model for solid tumors, especially non-vascularized micrometastases. The glucocorticoid prednisolone was tested for its influence on the release of TNF-alpha and the activity of PBMCs. RESULTS: It was shown that PBMCs, which were stimulated with a trifunctional bispecific antibody, BiUII, displayed an excellent antitumor activity, resulting in complete disintegration of the MTS. Also, it was demonstrated that prednisolone significantly reduced the release of TNF-alpha, without impairing the antitumor activity of BiUII-activated PBMCs. In contrast, unspecific killing was reduced, as demonstrated with an identical trAb (Bi48), which recognizes an antigen absent from the target cells. CONCLUSION: The in vivo application of bispecific antibodies for adjuvant tumor therapies may be limited by the manifest activation of immune effectors, accompanied by overwhelming cytokine release. Glucocorticoids, like prednisolone, may effectively reduce cytokine release without impairing the antitumor activity of trAb-activated immune cells. PMID- 16309223 TI - Protective effect of melatonin on human peripheral blood hematopoeitic stem cells against doxorubicin cytotoxicity. AB - BACKGROUND: The dose-limiting toxicity of doxorubicin on hematopoietic stem cells reduces the maximum benefit from this powerful drug. Melatonin may play a role in reducing this toxicity. MATERIALS AND METHODS: Melatonin at 10 microM was used while challenging human peripheral blood stem cells (PBSC) with doxorubicin (0.6 microM and 1 microM), and colony formation was used to evaluate the protective effect of melatonin. RESULTS: Melatonin was protective for the myeloid and erythroid series when given during or 1 hour after, but not before, doxorubicin, as measured by colony assay. This protection was independent from its antioxidant function as measured by 2', 7'-dichlodihydro-fluorescein diacetate and was selective for PBSC when compared to the MCF-7 cancer cell line. CONCLUSION: The results suggest the importance of the time sequence for melatonin administration to exert its protective effect in relation to doxorubicin treatment, as well as its protective effect on both erythroid and myeloid elements independent from its antioxidant function. PMID- 16309224 TI - E-cadherin and beta-catenin expression in normal, inflamed and cancerous gallbladder tissue. AB - BACKGROUND: Changes of the E-cadherin/beta-catenin complex during cell-cell interactions result in the loss of cell adhesion and may account for the ability of cancer cells to metastasize. Gallbladder carcinoma (GBC) can develop during chronic inflammation from normal tissue. The aim of this study was to investigate the expression of E-cadherin and beta-catenin in normal gallbladder mucosa, inflamed gallbladder tissue and GBC. MATERIALS AND METHODS: Tissue from 10 GBC, 10 chronic cholecystitis and 10 healthy gallbladders were used to evaluate the expression of E-cadherin and beta-catenin by immunohistochemistry. RESULTS: The beta-catenin membranous expression decreased between cholecystitis and malignant tissue, as well as between normal epithelium and carcinoma. The E-cadherin membranous expression was reduced in normal gallbladder epithelia compared to carcinoma and also from inflammation to GBC. The cytoplasmatic beta-catenin expression did not show any significant difference. Cytoplasmatic E-cadherin was significantly different from normal gallbladders to carcinomas and between normal tissue and inflammation. No significant difference of the nuclear P-catenin expression could be observed. E-cadherin was not detected intra-nuclear in any tissue. CONCLUSION: Significant differences of E-cadherin and beta-catenin were detected between normal, inflamed and cancerous tissues. These changes of the protein expressions and the associated loss of adhesive mechanisms might lead to a cancerous pathway in GBC. PMID- 16309225 TI - Modes of action of alpha-hederin and thymoquinone, active constituents of Nigella sativa, against HEp-2 cancer cells. AB - Our previous studies on active constituents of Nigella sativa have indicated that cell death induced by thymoquinone and alpha-hederin was dose- and time dependent, in a range of four cancer cell lines. Both compounds elicited necrosis and apoptosis with a higher incidence of the latter induced by thymoquinone. As HEp-2 human laryngeal carcinoma cells were the most susceptible, we sought to better understand the mechanisms involved by using buthionine sulfoximine (BSO), a selective inhibitor of glutathione (GSH) synthesis, to determine the importance of GSH in the apoptosis elicited, using cisplatin as internal standard. BSO significantly enhanced alpha-hederin- and cisplatin- mediated toxicity as assessed by the MIT assay, without changes in apoptosis or necrosis levels. Although the MTI assay did not indicate BSO potentiation of thymoquinone, apoptosis levels were significantly enhanced following this combination, without changes in necrosis. Thymoquinone and cisplatin significantly decreased GSH levels in a dose-dependent manner, with BSO pre-treatment synergistically depleting GSH levels in only thymoquinone- treated cells. As the caspase 3 inhibitor, Z-DEVD-fmk significantly decreased thymoquinone- and cisplatin-induced apoptosis, GSH depletion and caspase 3-activation mediate thymoquinone-induced apoptosis, in this cell line. PMID- 16309226 TI - Cancer of the uterine cervix is susceptible to anti-EGF-R antibody EMD 55,900 therapy. AB - BACKGROUND: Among a variety of cancer therapeutics targeting the epidermal growth factor receptor (EGF-R), monoclonal antibodies have shown some therapeutic potential in the treatment of various tumours of different entities. Nevertheless, several high EGF-R-expression carcinomas show no response to this treatment. Tumours of the uterine cervix represent a group, in which the response to anti-EGF-R treatment is hardly predictable, despite a relatively high expression of the receptor. MATERIALS AND METHODS: To assess the antitumour activity of anti-EGF-R monoclonal antibody EMD 55,900 in vivo, a series of experiments were performed in the nude mouse model using xenotransplanted primary carcinomas of the uterine cervix. RESULTS AND CONCLUSION: EMD 55,900 was found to be capable of inhibiting the growth of primary carcinomas of the uterine cervix at different stages of tumour development. The therapeutic response was not dependent on EGF-R expression solely, but also on the pre-treatment microvessel density. PMID- 16309227 TI - Cox-2 levels in canine mammary tumors, including inflammatory mammary carcinoma: clinicopathological features and prognostic significance. AB - Cyclo-oxygenase (Cox-2) plays an important role in mammary carcinogenesis, nevertheless, its role in canine mammary tumors, and particularly in inflammatory mammary carcinoma (IMC), is unknown. Tumor Cox-2 levels were analyzed by enzyme immunoassay, in post-surgical tumor homogenates of 129 mammary tumors (62 dysplasias and benign tumors, 57 malignant non-IMC and 10 IMC) from 57 female dogs. The highest Cox-2 values were detected in the IMC group. In non-IMC malignant tumors, high values of Cox-2 were related to skin ulceration (p < 0.001) and tumor size (p < 0.001). The follow-up study revealed that high Cox-2 levels were related with recurrence (p = 0.002), metastases (p < 0.001), disease free survival (p < 0.001) and overall survival (p < 0.001). This study demonstrates an association between intra-tumor Cox-2 levels and poor prognosis. The high levels found in IMC cases could indicate a special role of Cox-2 in the inflammatory phenotype and open the possibility of additional new therapeutic approaches in this special type of mammary cancer in humans and dogs. PMID- 16309228 TI - 4-OH tamoxifen does not interfere with bicalutamide inhibitory effects on human prostatic cancer cells in vitro. AB - BACKGROUND: The present study was aimed at investigating the effects of the co administration of 4-OH Tamoxifen (4HT) and bicalutamide (BIC) on the human prostate cancer cell line LNCaP in vitro. MATERIALS AND METHODS: The LNCaP FGC prostate cancer cell line was grown in vitro within a three-dimensional matrix formed by collagen gel under dihydrotestosterone (DHT) (0.1 nM) stimulation. Cells were incubated in the presence either of BIC at escalating concentrations (1 nM; 100 nM; 10 microM) or 4HT Tamoxifen (10 nM; 100 nM) or both compounds at the different concentrations studied. The cells were incubated for 144 hours, and growth was evaluated in non trypsinised cells by crystal violet vital dye staining assay. RESULTS: BIC appeared to exert a dose-dependent inhibitory action, with the maximum inhibitory effect achieved by the 10 microM concentration. A comparable inhibitory effect was also observed after exposure to 4HT at both doses tested. No statistically significant interference was observed when BIC was combined with 4HT. CONCLUSION: 4HT, even at the higher concentration employed here, showed no major interference with the inhibitory effects of BIC. PMID- 16309229 TI - Growth inhibitory effect of doxazosin on prostate and bladder cancer cells. Is the serotonin receptor pathway involved? AB - Doxazosin, an alpha1-adrenoceptor antagonist, is used for the treatment of benign prostatic hyperplasia (BPH) and hypertension. Alpha-adrenoceptor antagonists also inhibit growth and induce apoptosis in malignant prostatic cells. The apoptotic activity is independent of their capacity to antagonize alpha-adrenoceptors. The effect of doxazosin on the growth of prostate and bladder cancer cell lines was assessed and whether the growth inhibitory effect of doxazosin on prostate cancer cells is serotonin (5-hydroxtryptamine; 5HT)-dependent was investigated. MATERIALS AND METHODS: PC3 (androgen-independent prostate cancer) and HT1376 (grade III transitional cell carcinoma) cells were plated. The cells were incubated with doxazosin. After 72 h, cell viability was assessed (crystal violet assay). Studies were also performed after incubating the PC3 cells with 5HT or 5HT(1B) agonists for a short duration, followed by the addition of doxazosin. Cell viability was assessed at 72 h. RESULTS: Doxazosin caused a dose-dependent inhibition of PC3 and HT1376 cell growth with a maximum inhibition of 80% (n=12, p < 0.0001) and 91% (n=12, p < 0.0001), respectively, at a concentration of 10( 4)M, at 72 h. Incubation of PC3 cells with 5HT or 5HT(1B) agonist, followed by addition of doxazosin, increased the percent of viable cells as compared to when the cells were treated with doxazosin alone. CONCLUSION: Doxazosin significantly inhibited prostate (PC3) and bladder cancer (HT1376) cell growth. Furthermore, prior incubation of PC3 cells with 5HT or 5HT(1B) agonist increased cell viability as compared to treatment with doxazosin alone. These findings may be related to the similarity between subtype 1 serotonin and adrenergic receptors. The effect of alpha1-adrenoceptor antagonists on tumour cell growth merits further investigation. PMID- 16309230 TI - Expression analysis of a subset of coregulators and three nuclear receptors in human colorectal carcinoma. AB - In human colorectal tissue samples, the gene expressions of 4 coactivators, p300, pCAF, TIF-2 and TRAP 220, and 7 corepressors, N-CoR, REA, MTA1, MTA1L1, HDAC1, HDAC2 and HDAC3, linked to estrogen receptors (ER), were revealed by traditional RT-PCR. Cofactors ERalpha, ERbeta and ERRalpha mRNA levels were then measured in 40 tumor tissue samples matched with respective normal mucosa by real-time PCR. The decline of mRNA levels of all coactivators and the increase of NCoR, HDAC1, HDAC2 and MTA1 were observed from normal to tumor tissue, whereas REA, HDAC3 and MTA1L1 expressions were similar in both tissue compartments. The gene expression of ERbeta correlated with those of p300, TIF-2 and REA in normal mucosa, and with that of REA in tumor tissue only. No association was found between ERalpha and coregulators and between each coregulator and different clinical parameters. Our findings suggest that the co-induction of ERbeta and some cofactors may play an important role during the development of human colorectal carcinoma. PMID- 16309231 TI - Proposal of a new grading system for malignant fibrous histiocytomas. AB - The proposed grading system for malignant fibrous histiocytomas (MFH) comprises 3 grades of malignancy. Analogous to other grading systems, the system includes the factors of mitotic rate and necrosis. In addition to these two factors, the concept of cellularity was included. The prognostic relevance of the grading systems published by Costa, Coindre, van Unnik, Pezzi and Tsujimoto as well as the grading system proposed by the present study was tested on 161 MFH. The results showed that all grading systems tested produced clearly significant differences (p < 0.01) with regard to the survival estimated for patients with various grades of malignancy. These results revealed the superiority of systems that use 3 grades of malignancy over a 2-grade classification. The proposed grading system yielded a lower percentage of grade II tumours (37%) than the grading systems of Coindre (60%) and van Unnik (70%). In the multivariate analysis of all grading systems, the proposed grading system was the only one to show prognostic relevance (p < 0. 05). PMID- 16309232 TI - Molecular changes in PDEGF and bFGF in malignant melanomas in relation to the stromal microenvironment. AB - BACKGROUND: Aberrant expression of either growth factors or growth factor receptors by stromal cells can be an important factor promoting the growth of solid tumours. It may also affect differentiation of malignant cells and support tumour spread. The aim of the present study was to investigate the hypothesis that basic-fibroblast growth factor (bFGF) and platelet-derived growth factor (PDEGF) may be involved in tumour-stromal microenvironment interactions in primary malignant melanomas. MATERIALS AND METHODS: PDEGF and bFGF expression in malignant cells and surrounding stromal elements was assessed using indirect immunohistochemistry. RESULTS: It was confirmed that PDEGF can be involved in the reciprocal interactions between tumour cells and stroma, including aberrant angiogenesis. Interestingly, bFGF was present both in malignant melanoma lesions and benign nevi accompanied by different intracellular localisation of the protein, suggesting its implication in regulation of nevus cell proliferation and maturation. CONCLUSION: The present results suggest that bFGF and PDEGF participate in malignant melanoma progression. PMID- 16309233 TI - Insulin-like growth factor-I receptor expression and localization in benign and malignant rat mammary tumors. AB - The aim of this study was to investigate the expression and localization of the insulin growth factor type 1 receptor (IGF-IR) in malignant and benign mammary tumors induced in rats by N-nitroso-N-methylurea (NMU) and its correlation with histopathology and hormone dependence. Also, protein tyrosine kinase activities (PTKs) were analyzed in order to study the activation of the intracellular cascade. The results showed that IGF-IR is present in NMU tumors (analyzed by binding assay and Western blot), that a variable content is expressed in tumors that continued growing post-ovariectomy (OVX) of rats, and that it is undetectable in tumors that regressed post-OVX. IGF-IR was principally localized (by immunohistochemistry) in the epithelial cells of malignant tumors and in the fibrous cells of benign ones. Also, a significantly lower expression of both cytosolic and microsomal PTKs were found in benign tumors. Our results suggest a different expression and role of IGF-IR in benign and malignant tumors. PMID- 16309234 TI - Endothelial cell survivin is involved in the growth of ovarian endometriotic cysts. AB - BACKGROUND: The aim of the present study was to evaluate microvessel density (MVD) in the cellular layers of ovarian endometriomata, with particular interest in the relationship with VEGF and survivin expressions by endothelial cells and with the diameter of the cysts. MATERIALS AND METHODS: MVD and VEGF and survivin endothelial cell expressions were evaluated in 26 ovarian endometriotic cysts and correlated with the cyst diameter. RESULTS: The mean MVD was higher in the inner specialized stroma of ectopic endometrium than in the outer fibrous capsule, but only in the fibrous capsule was MVD correlated with endothelial cell VEGF and survivin expressions as well as with the cyst diameter. CONCLUSION: The diameter of ovarian endometriotic cysts seems to be related to the angiogenic process involving the outer fibrous capsule, and not the inner specialized stroma of ectopic endometrium, since only in the capsule are vessels stimulated to proliferate by VEGF and protected from apoptosis by survivin, and their density is correlated to cyst diameter. PMID- 16309235 TI - Influence of hMLH1 methylation, mismatch repair deficiency and microsatellite instability on chemoresistance of testicular germ-cell tumors. AB - BACKGROUND: Cisplatin-based chemotherapy can cure more than 80% of metastatic germ-cell testicular tumors (GCTs). The response to cisplatin-based chemotherapy has been related to Microsatellite Instability (MSI), which is caused by genetic or epigenetic changes in genes of the DNA Mismatch Repair (MMR) pathway. PATIENTS AND METHODS: We investigated 15 refractory and 36 chemosensitive GCTs for immunohistochemical loss of hMLH1, hMSH2 and hMSH6 protein expressions, in conjunction with hMLH1 gene methylation and MSI of GCTs, with a complete follow up. RESULTS: A loss of either of the MMR protein expressions was detected in 14 cases (27.5%). Pathological hMLH1 protein expression was seen in 10 cases (19.6%). hMLH1 methylation was found in 11 cases (21.60%) and was highly correlated with loss of hMLH1 expression (p < 0.0001) and with immunohistochemically-detected MMR deficiency (p = 0.0005). MSI was found in 16 cases (31.4%). There was no correlation between hMLH1 methylation and MSI. Neither hMLH1 methylation status, nor MSI correlated with any of the clinicopathological parameters investigated (tumor stage, histology, resistance to systemic treatment). CONCLUSION: hMLH1 gene methylation was detected as a common alteration in GCTs, and correlated with the loss of hMLH1 protein expression (p < 0.0001). Neither hMLH1 gene methylation, MMR deficiency, nor MSI showed a relationship with the relevant clinicopathological parameters. PMID- 16309236 TI - Synergistic cytotoxicity of artemisinin and sodium butyrate on human cancer cells. AB - BACKGROUND: Butyric acid is a short chain fatty acid produced by large bowel bacterial flora. It serves as an antiinflammatory agent and nutrient for normal colon cells. Butyric acid has also been shown to induce apoptosis in colon and many other cancer cells. Artemisinin is a compound extracted from the wormwood Artemisia annua L. It has been shown to selectively kill cancer cells in vitro and to be effective in treating animal and human cancer. We and others have found that the artemisinin analog, dihydroartemisinin (DHA), kills cancer cells by apoptosis. In the present study, the efficacy of a combined treatment of DHA and butyric acid at low doses in killing cancer cells was investigated. MATERIALS AND METHODS: Molt-4 cells (a human lymphoblastoid leukemia cell line) and freshly isolated human lymphocytes, cultured in complete RPMI-1640 medium, were first incubated with 12 microM of human holotransferrin at 37 degrees C in a humid atmosphere of 5% CO2 for one hour to enhance the iron concentration in the cells. Cells from each cell type were then divided into 20 flasks. These flasks were grouped into four sets of five cultures each. Zero, 5, 10 or 20 microM of DHA was added, respectively, to these sets and the cells were incubated at 37 degrees C for one hour. Zero, 1, 5, 10, or 20 mM of sodium butyrate was then added to the five cultures of each set, respectively. Thus, the treatments involved a combination of 4 doses of DHA and 5 doses of sodium butyrate. The cells were counted immediately before the addition of DHA, and at 24 and 48 hours after the addition of sodium butyrate. RESULTS: DHA alone at the 24-hour time-point and 20 microM concentration significantly reduced the number of Molt-4 cells in the culture by approximately 40% (p < 0.001, compared to non-treated control), whereas it did not significantly affect the number of normal human lymphocytes. Similarly, 1 mM sodium butyrate alone at 24 hours reduced the number of Molt-4 cells by approximately 32% (p < 0.001, compared to non-treated control), without significantly affecting normal human lymphocytes. The combination of 20 microM DHA and 1 mM sodium butyrate killed all Molt-4 cells at the 24-hour time-point and did not significantly affect lymphocytes. CONCLUSION: DHA in combination with butyric acid acts synergistically at low doses. The combination may provide a less toxic, inexpensive and effective cancer chemotherapy. PMID- 16309237 TI - Growth inhibition of cancer cells by an active metabolite of a novel vitamin D prodrug. AB - Active vitamin D compounds have been developed that maintain antiproliferative properties with low calcemic activity. BCI-210, a novel vitamin D pro-drug developed in our laboratory, is activated through side chain hydroxylation and possesses lower calcemic activity than calcitriol. The human hepatoma cell line (HepG2) was used to produce an active metabolite, which was characterized and identified as 27-hydroxy-BCI-210. We compared the ability of 27-OH-BCI-210 with calcitriol to inhibit proliferation of prostate (LNCaP), and breast (MCF-7) cancer cells. Cells were plated in multi-well plates and incubated with vehicle or vitamin D compounds for 6 days, after which the cell numbers were determined by a colorimetric assay. 27-OH-BCI-210 produced a dose-dependent growth inhibition, although a concentration five-fold greater than calcitriol was required to produce equivalent inhibition. We also examined the antiproliferative activity of 27-OH-BCI-210 in combination with chemotherapeutic drugs. With genistein and doxorubicin, 27-OH-BCI-210 produced synergistic inhibition of proliferation of LNCaP and MCF-7 cells. These synergistic interactions suggest the potential clinical utility of 27-OH-BCI-210 in the treatment of prostate and breast tumors. PMID- 16309238 TI - Flavopiridol, an inhibitor of cyclin-dependent kinases, induces growth inhibition and apoptosis in bladder cancer cells in vitro and in vivo. AB - Flavopiridol is a semi-synthetic flavone analog of the alkaloid, rohitukine, a compound from an Indian tree, Dysoxylum binectariferum. It has been shown to inhibit cyclin-dependent kinases (CDKs), causing cell cycle arrest and growth inhibition. Flavopiridol is reported to have cytotoxic activity against a wide range of cancer cell lines and has demonstrated its efficacy in several clinical trials. Flavopiridol seems a well-suited potential new agent for the treatment of bladder cancer. We, therefore, evaluated whether flavopiridol inhibits growth and induces apoptosis in bladder cancer cells and additionally examined the toxicity and efficacy of this drug in vivo in a rat bladder cancer model. The in vitro experiments showed an IC20 of 50-100 nM in all cell lines tested. However, there was a difference in the response with regard to the grading of the tumor cells at higher doses. The IC50 was found to be 150-350 nM in the well-differentiated RT4 and RTI12 cell lines after treatment with flavopiridol, in comparison to a IC50 of 1000 nMfor the poorly-differentiated cell lines T24 and SUP. After exposure to flavopiridol, all tumor cell lines underwent significant apoptosis in comparison to untreated cells, beginning at a dose of 50 nM flavopiridol. At high concentrations (500 nM) of flavopiridol, 80-90% of all cells showed severe apoptotic alterations. The treatment of rat urinary bladder cancer with flavopiridol demonstrated the best efficacy with an intermittent treatment of 0.1 mg/kg, 3 times weekly over a total of 3 weeks, resulting in 7/12 animals tumor free and a trend for the remaining tumors to have lower stage and grade. There seems to be a small advantage in intermittent versus daily application of flavopiridol. In summary, our results indicated that flavopiridol could be a useful therapeutic agent for bladder cancer, inhibiting tumor growth, malignant progression and inducing apoptosis. PMID- 16309239 TI - Adenovirus-mediated transfer of FRNK augments drug-induced cytotoxicity in cultured SCCHN cells. AB - BACKGROUND: Focal adhesion kinase (FAK), which is overexpressed in many human epithelial cancers, regulates cell cycle progression, cellular migration, invasion and survival. MATERIALS AND METHODS: In order to determine if inhibiting FAK activity augments drug-induced cytotoxicity in transformed epithelial cells from the head and neck region (SCCHN cells), cells were transfected with a recombinant adenovirus causing overexpression of FRNK a dominant negative inhibitor of FAK Results: When SCCHN cells (SCC25 and RPMI 2650) were transfected with Ad-FRNK there was a decrease in autophosphorylation of FAK, coincident with a large increase in FRNK expression. Ad-FRNK and cytotoxic drugs were more effective in reducing cell viability and increasing apoptosis then Ad-FRNK alone or drugs alone. Transfection with Ad-FRNK also led to substantially decreased migration of cultured SCCHN cells. These results indicate that FAK may be a good target for anticancer therapy. PMID- 16309240 TI - Combined radiation sensitizing and anti-angiogenic effects of ionizing radiation and the protease inhibitor ritonavir in a head and neck carcinoma model. AB - Ritonavir, a protease inhibitor, has been successfully applied in the treatment of HIV infection. Reports of dramatic improvement of AIDS-related cancers, such as primary central system lymphoma after radiation therapy as well as Kaposi's sarcoma, led to the recent discovery of the "non viral" antitumor activity of HIV protease inhibitors. This study was designed to detect the antitumor effect of Ritonavir when combined with ionizing radiation both in vitro and in vivo in the HEP-2 head and neck carcinoma model. Inhibition of tumor growth was observed when mice were treated with Ritonavir alone and this effect was enhanced when combined with ionizing radiation. No adverse effect or significant toxicity in the hosts' body weights was seen between the different treatment and control groups throughout the experiments. A marked antiproliferation effect of the combination was observed in vitro. A marked reduction of angiogenesis was detected within the tumor sections from the Ritonavir combined with irradiation group as compared with the Ritonavir or irradiation alone groups. Western blot analysis showed that apoptosis was induced by an increased expression of Bax and decreased expression of Bcl-2 after treatment with Ritonavir and ionizing radiation. Thus, the antitumor effect of the latter combination is associated with the enhancement of radiation-induced apoptosis and inhibition of angiogenesis. These data suggested that Ritonavir could clinically improve the tumor response to radiation therapy, especially in head and neck carcinoma. PMID- 16309241 TI - Head and neck carcinoma cell lines are sensitive to vinflunine in vitro. AB - BACKGROUND: New treatment models are needed for the management of advanced stage head and neck carcinomas (HNSCC). Vinflunine is a novel vinca alkaloid which has shown remarkable improvement in efficiency and reduced side-effects in both in vitro and in vivo studies. Better locoregional control and prevention of distant metastasis in advanced HNSCC would be welcome and, with this new efficient compound, these might be expected. The purpose of this study was to evaluate the sensitivity of HNSCC cell lines to vinflunine in vitro. MATERIALS AND METHODS: Nine recently established cell lines were used: UT-SCC-1A, -2 and -33 (oral cavity SCC), UT-SCC-40 (tongue SCC), UT-SCC-9, -11, -19A, -29 and -34 (laryngeal SCC). Cells in midlogarithmic growth were used in the 96-well plate clonogenic assay. Vinflunine was tested using two 96-well plates for each concentration, the experiments were repeated 4-6 times. The plating efficiency was calculated, the survival data was fit to the linear quadratic equation F = exp[-(alphaD + betaD2)] and IC50 corresponding to the mean inactivation dose, were obtained from the dose response curves. RESULTS: All tested cell lines were sensitive to vinflunine in nanomolar concentrations. The IC50 values of the same cell lines to vinorelbine were only slightly lower, with means of 0.96 nM and 0.73 nM, respectively. CONCLUSION: These results indicate that the new vinca alkaloid, vinflunine, could be useful in the clinical setting, and in vitro testing in combined modality models, such as concurrent chemoradiation, is warranted. PMID- 16309242 TI - Absence of interaction between doppel and GFAP, Grb2, PrPc proteins in human tumor astrocytic cells. AB - BACKGROUND: The doppel protein (Dpl) is a newly recognized cellular prion protein (PrP(C))-like molecule encoded by a novel gene locus, PRND, located on the same chromosomal region of the PrP(C) coding gene. Recently, Dpl was shown to be aberrantly expressed in astrocytic tumor specimens and in astrocytoma-derived cell lines, showing a peculiar cytoplasmic localization. Here, Dpl interactions with some of the prion-interacting proteins were studied. In particular, whether the tumor astrocytic environment is suitable for doppel interaction with GFAP and Grb2 proteins, as well as with the PrPC protein itself was investigated. MATERIALS AND METHODS: In order to verify our hypothesis, an innovative mammalian two-hybrid system and co-immunoprecipitation assays were employed. RESULTS: The results reported the absence of protein interactions. Our findings provided evidence that, in our astrocytoma cell-based model, Dpl does not share with PrP(C) the ability to interact with GFAP and Grb2. CONCLUSION: Identifying Dpl ligands may provide new insights into the involvement of Dpl in astrocytoma tumor progression. PMID- 16309244 TI - Nano-to-microscale mechanical switches and fuses mediate adhesive contacts between leukocytes and the endothelium. PMID- 16309245 TI - Modeling of the spread of an effluent and the overturning length scale near an underwater source in the northern Adriatic. PMID- 16309246 TI - A perspective on the slow vacuolar channel in vacuoles from higher plant cells. PMID- 16309247 TI - Variable selection and interpretation in structure-affinity correlation modeling of estrogen receptor binders. AB - A computational approach for the identification and investigation of correlations between a chemical structure and a selected biological property is described. It is based on a set of 132 compounds of known chemical structures, which were tested for their binding affinities to the estrogen receptor. Different multivariate modeling methods, i.e., partial least-squares regression, counterpropagation neural network, and error-back-propagation neural network, were applied, and the prediction ability of each model was tested in order to compare the results of the obtained models. To reduce the extensive set of calculated structural descriptors, two types of variable selection methods were applied, depending on the modeling approach used. In particular, the final partial least-squares regression model was built using the "variable importance in projection" variable selection method, while genetic algorithms were applied in neural network modeling to select the optimal set of descriptors. A thorough statistical study of the variables selected by genetic algorithms is shown. The results were assessed with the aim to get insight to the mechanisms involved in the binding of estrogenic compounds to the receptor. The variable selection on the basis of genetic algorithm was controlled with the test set of compounds, extracted from the data set available. To compare the predictive ability of all the optimized models, a leave-one-out cross-validation procedure was applied, the best model being the nonlinear neural network model based on error back propagation algorithm, which resulted in R2= 92.2% and Q2= 70.8%. PMID- 16309249 TI - Chirality of toroidal molecular graphs. AB - Symmetry properties of a class of toroidal molecular graphs, arising as covers of certain bipartite cubic Cayley graphs of dihedral groups, are studied. Although these symmetries make all vertices and all edges indistinguishable, they imply intrinsic chirality. PMID- 16309248 TI - Complex kinetics of the electron transfer between the photoactive redox label TUPS and the heme of cytochrome c. AB - The photoinduced covalent redox label 8-thiouredopyrene-1,3,6-trisulfonate (TUPS) has been attached to two lysine residues (K8 and K39) at opposite sides of horse heart cytochrome c, as well as to cysteines, at the same positions, introduced by site-directed mutagenesis. Electron transfer between TUPS and the heme of cytochrome c deviates from the expected monoexponential kinetic behavior. Neither the overall rate nor the individual exponential components of electron transfer, as followed by kinetic absorption spectroscopy, correlate with the length of the covalent link connecting the dye with the protein. Molecular dynamics calculations show that TUPS can approach the protein surface and occupy several such positions. This heterogeneity may explain the multiexponential electron transfer kinetics. The calculated optimal electron-transfer pathways do not follow the covalent link but involve through space jumps from the dye to the protein moiety, effectively decoupling the length of the covalent link and the electron-transfer rates. PMID- 16309250 TI - Metabotropic glutamate receptor-mediated currents at the climbing fiber to Purkinje cell synapse. AB - Different forms of synaptic plasticity in the cerebellum expressed at the synapses onto Purkinje cells (PCs) are mediated by membrane metabotropic glutamate receptors (mGluRs). There are three main mGluR groups with a total of 8 subtypes. Although mGluRs are also found at the climbing fiber (CF) to PC synapses, the distribution and biological activity of their types is not well known. Using whole cell patch-clamp recordings from PCs in rat cerebellar slices with inhibitors of ionotropic receptors and glutamate uptake blockers we demonstrate a complex pharmacology of currents obtained by CF stimulation. The mGluR1 specific antagonist CPCCOEt in a group of cells suppressed this response, but in a similar number of other cells it induced a potentiating effect. It was found that a switch between these two biopharmaceutical effects might occur with age. PMID- 16309251 TI - Staphylococcus aureus bicomponent gamma-hemolysins, HlgA, HlgB, and HlgC, can form mixed pores containing all components. AB - Staphylococcal gamma-hemolysins are bicomponent toxins forming a protein family with leucocidins and alpha-toxin. Two active toxins (AB and CB) can be formed combining one of the class-S components, HlgA or HlgC, with the class-F component HlgB. These two gamma-hemolysins form pores with marked similarities to alpha toxin in terms of conductance, nonlinearity of the current-voltage curve, and channel stability in the open state. AB and CB pores, however, are cation selective, whereas alpha-toxin is anion-selective. gamma-Hemolysins' pores are hetero-oligomers formed by three or four copies of each component (indicated as 3A3B and 3C3B or 4A4B and 4C4B). Point mutants located on a beta-strand of the class-S component that forms part of the protomer-protomer contact region can prevent oligomer assembly. Interestingly, these mutants inhibit growth of pores formed not only by their natural components but also by nonstandard components. This lead to the hypothesis that mixed ABC pores could also be formed. By studying the conductance of pores, assembled in the presence of all three components (in different ratios), it was observed that the magnitudes expected for mixed pores were, indeed, present. We conclude that the gamma hemolysin/leucocidin bicomponent toxin family may form a larger than expected number of active toxins by cross-combining various S and F components. PMID- 16309252 TI - Development and validation of empirical force field parameters for netropsin. AB - The netropsin molecule preferentially binds to the four consecutive A.T base pairs of the DNA minor groove and could therefore inhibit the expression of specific genes. The understanding of its binding on a molecular level is indispensable for computer-aided design of new antitumor agents. This knowledge could be obtained via molecular dynamics (MD) and docking simulations, but in this case appropriate force field parameters for the netropsin molecule should be explicitly defined. Our parametrization was based on the results of quantum chemical calculations. The resulting set of parameters was able to reproduce bond lengths, bond angles, torsional angles of the ab initio minimized geometry within 0.03 A, 3 deg and 5 deg, respectively, and its vibrational frequencies with a relative error of 4.3% for low and 2.8% for high energy modes. To show the accuracy of the developed parameters we calculated an IR spectrum of the netropsin molecule using MD simulation and found it to be in good agreement with the experimental one. Finally, we performed a 10 ns long MD simulation of the netropsin-DNA complex immersed in explicit water. The overall complex conformation remained stable at all times, and its secondary structure was well retained. PMID- 16309253 TI - GISAXS view of vanadium/cerium oxide thin films and influence of lithium intercalation. AB - An examination of structural modifications, induced by mixing vanadium and cerium oxides and by the introduction of lithium in vanadium and mixed vanadium/cerium oxide films, was performed using synchrotron sourced grazing incidence small angle X-ray scattering. Samples were sol-gel-derived films, deposited by a dip coating technique. An analysis of the scattering data, acquired by a two dimensional detection system, is based on the comparison of the surface and bulk characteristics of the film. The trend of estimated structural modifications is supported by the results of previous investigations on a different length scale, performed by atomic force microscopy. PMID- 16309254 TI - Biomedical simulation of heat transfer in a human heart. AB - Theory and practical experiences from numerical simulations of heat transfer in the field of medicine are presented in this paper. The cooling of a human heart during surgery was taken as an illustrative example. All phases of the simulation process are described starting with the construction of an irregularly shaped 3 dimensional model. The mathematical model is based on diffusion and Navier-Stokes equations. The system of partial differential equations is solved by finite difference approximation using an explicit time-stepping scheme to obtain the time evolution of the solution for the complete simulated interval, which is typically 1 h. A typical domain is composed of several million voxels; therefore, the program was parallelized to speed up the simulation. A speed-up of 8.2 was obtained on 16 processors in a Linux cluster. PMID- 16309255 TI - Reaction between ellagic acid and an ultimate carcinogen. AB - The reaction coordinate between a typical ultimate carcinogen benzo[a]pyrene-7,8 diol-9,10-epoxide (BPDE) and ellagic acid, a proven chemopreventive agent active against cancers caused by polycyclic aromatic hydrocarbons (PAHs), was examined by density functional theory (DFT) and semiempirical MO calculations, and activation energy was calculated. The effect of a polar environment was included using Tomasi and the Langevin dipoles methods. The calculated BPDE/ellagic acid reaction free energy of activation is found to be in decent agreement with experimental data [Sayer, J. M. et al. J. Am. Chem. Soc. 1982, 104, 5562-5564]. This work sheds light on the mechanism of action of ellagic acid. Quantum chemical calculations of this kind are valuable for the design of ellagic acid derivatives with even lower activation energy and increased reactivity toward ultimate carcinogens as well as controlled reactivity toward DNA. PMID- 16309256 TI - Molecular dynamics integration meets standard theory of molecular vibrations. AB - An iterative SISM (split integration symplectic method) for molecular dynamics (MD) integration is described. This work explores an alternative for the internal coordinate system prediction in the SISM introduced by JaneZic et al. (J. Chem. Phys. 2005, 122, 174101). The SISM, which employs a standard theory of molecular vibrations, analytically resolves the internal high-frequency molecular vibrations. This is accomplished by introducing a translating and rotating internal coordinate system of a molecule and calculating normal modes of an isolated molecule only. The Eckart frame, which is usually used in the standard theory of molecular vibrations as an internal coordinate system of a molecule, is adopted to be used within the framework of the second order generalized leapfrog scheme. In the presented MD integrator the internal coordinate frame at the end of the integration step is predicted halfway through the integration step using a predictor-corrector type iterative approach thus ensuring the method's time reversibility. The iterative SISM, which is applicable to any system of molecules with one equilibrium configuration, was applied here to perform all-atom MD simulations of liquid CO2 and SO2. The simulation results indicate that for the same level of accuracy, this algorithm allows significantly longer integration time steps than the standard second-order leapfrog Verlet (LFV) method. PMID- 16309257 TI - Electronic coupling effects on photoinduced electron transfer in carotene porphyrin-fullerene triads detected by time-resolved EPR. AB - Photoinduced charge separation and recombination in a carotenoid-porphyrin fullerene triad C-P-C60 (Bahr et al., 2000) have been followed by time-resolved electron paramagnetic resonance. The electron-transfer process has been characterized in a glass of 2-methyltetrahydrofuran and in the nematic phase of two uniaxial liquid crystals (E-7 and ZLI-1167). In all the different media, the molecular triad undergoes two-step photoinduced electron transfer, with the generation of a long-lived charge-separated state (C*+-P-C60*-), and charge recombination to the triplet state, localized in the carotene moiety, mimicking different aspects of the photosynthetic electron-transfer process. The magnetic interaction parameters have been evaluated by simulation of the spin-polarized radical pair spectrum. The weak exchange interaction parameter (J = +1.7 +/- 0.1 G) provides a direct measure of the dominant electronic coupling matrix element V between the C*+-P-C60*- radical pair state and the recombination triplet state 3C P-C60. Comparison of the estimated values of V for this triad and a structurally related triad differing only in the porphyrin bridge (octaalkylporphyrin vs tetraarylporphyrin) explains in terms of an electronic coupling effect the approximately 6-fold variation of the recombination rate induced by the modification of the porphyrin bridge as derived by kinetic experiments (Bahr et al., 2000). PMID- 16309258 TI - Entrapment of a weak polyanion and H+/Na+ exchange in confined polyelectrolyte microcapsules. AB - An approach for the entrapment of a polyanion by polyelectrolyte microcapsules is reported. It is based on a reversal changing of microcapsule wall permeability from neutral to basic pH. Polyelectrolyte microcapsules were templated on latex (polystyrene) particles by the layer-by-layer adsorption of oppositely charged polymers of sodium poly(styrene sulfonate) and poly(allylamine hydrochloride), followed by core removal using tetrahydrofuran. In alkaline conditions, the microcapsules swell and become permeable for polymers. During encapsulation, the addition of salt ions increases the amount of the polymer encapsulated and contributes to its protonation because of redistribution of H+ ions across a semipermeable microcapsule wall. The redistribution of small ions across the microcapsule wall was tuned by adding salt according to the Donnan equilibrium and was characterized by H+ sensitive dyes. PMID- 16309259 TI - High-density hydration layer of lysozymes: molecular dynamics decomposition of solution scattering data. AB - A characterization of the physical properties of protein hydration water is critical for understanding protein structure and function. Recent small-angle X ray and neutron scattering data indicate that the density of water on the surface of lysozyme is significantly higher than in bulk water. Here, we provide an interpretation of the scattering results using a molecular dynamics simulation, which allows us to make quantitative predictions about density variations in the first hydration shell. The perturbation relative to bulk water involves statistically significant changes in the average water structure in the first hydration layer. The water density in the first hydration shell is increased by 5% with respect to the bulk. In regions of higher water density, the water dipoles align more parallel to each other and the number of hydrogen bonds per water molecule is higher. Increased water density is found for water molecules interacting with hydrogen and carbon atoms in the backbone or with nonpolar or negatively charged side-chain groups. PMID- 16309260 TI - Symplectic molecular dynamics simulations on specially designed parallel computers. AB - We have developed a computer program for molecular dynamics (MD) simulation that implements the Split Integration Symplectic Method (SISM) and is designed to run on specialized parallel computers. The MD integration is performed by the SISM, which analytically treats high-frequency vibrational motion and thus enables the use of longer simulation time steps. The low-frequency motion is treated numerically on specially designed parallel computers, which decreases the computational time of each simulation time step. The combination of these approaches means that less time is required and fewer steps are needed and so enables fast MD simulations. We study the computational performance of MD simulation of molecular systems on specialized computers and provide a comparison to standard personal computers. The combination of the SISM with two specialized parallel computers is an effective way to increase the speed of MD simulations up to 16-fold over a single PC processor. PMID- 16309261 TI - Mathematical modeling of the myosin light chain kinase activation. AB - The mathematical model presented here describes the interactions among Ca2+, calmodulin (CaM), and myosin light chain kinase (MLCK) and consists of a kinetic scheme taking into account 7 reactions instead of 12 as proposed previously. We derive a system of 5 nonlinear ordinary differential equations. Solving it yields the prediction of active MLCK as a function of [Ca2+] whereby the active MLCK is defined to be proportional to the Ca4CaM.MLCK complex concentration. The model predictions are compared with other theoretical and experimental predictions of active MLCK as well as with the results of our previously proposed complex model. PMID- 16309262 TI - Mathematical modeling of the relation between myosin phosphorylation and stress development in smooth muscles. AB - In this paper the 4-state latch bridge model proposed by Rembold and Murphy is expanded; first by incorporation of the analytical expression of Ca2+ dependent MLCK activation from the work of Kato et al. and second, by inclusion of the myosin dephosphorylation based on the Michaelis-Menten kinetics. The analysis of the proposed model and the comparison with the original model results as well as with the experimental data is presented. The model is able to predict the steady state isometric stress and the myosin phosphorylation in dependence on steady cytosolic [Ca2+] as well as the temporal evolution of the system in dependence on the input Ca2+ signal in the form of biphasic transient, whereby our model results are in several aspects in better agreement with experimental observations. PMID- 16309263 TI - Probabilistic kinetic model of slow oxidation of low-density lipoprotein. 3. Hydroperoxide-free initiation. AB - A theoretical model is presented that explains slow copper-induced oxidation of low-density lipoprotein in the systems free of seeded hydroperoxydes. The model is based on the probabilistic kinetic theory, modified to take into account different radical generation rates by oxidized and reduced forms of the metal ions. It is shown that the initiation and progression of the LDL oxidation can take place in any LDL dispersion by metal-induced oxidation of alpha-tocopherol, one of the constituents of LDL, and formation of slowly reacting alpha tocopheroxyl radical. Selected values of several adjustable parameters define actual temporal profiles of the quantities defining the oxidation process. PMID- 16309264 TI - Exploring the local conformational space of a membrane protein by site-directed spin labeling. AB - Molecular modeling based on a hybrid evolutionary optimization and an information condensation algorithm, called GHOST, of spin label ESR spectra was applied to study the structure and dynamics of membrane proteins. The new method is capable of providing detailed molecular information about the conformational space of the spin-labeled segment of the protein in a membrane system. The method is applied to spin-labeled bacteriophage M13 major coat protein, which is used as a model membrane protein. Single cysteine mutants of the coat protein were labeled with nitroxide spin labels and incorporated in 1,2-dioleoyl-sn-glycero-3 phosphocholine (DOPC) bilayers. The new computational method allows us to monitor distributions of local spatial constraints and molecular mobility, in addition to information about the location of the protein in a membrane. Furthermore, the results suggest that different local conformations may coexist in the membrane protein. The knowledge of different local conformations may help us to better understand the function-structure relationship of membrane proteins. PMID- 16309265 TI - Speeding up a genetic algorithm for EPR-based spin label characterization of biosystem complexity. AB - Complexity of biological systems is one of the toughest problems for any experimental technique. Complex biochemical composition and a variety of biophysical interactions governing the evolution of a state of a biological system imply that the experimental response of the system would be superimposed of many different responses. To obtain a reliable characterization of such a system based on spin-label Electron Paramagnetic Resonance (EPR) spectroscopy, multiple Hybrid Evolutionary Optimization (HEO) combined with spectral simulation can be applied. Implemented as the GHOST algorithm this approach is capable of handling the huge solution space and provides an insight into the "quasicontinuous" distribution of parameters that describe the biophysical properties of an experimental system. However, the analysis procedure requires several hundreds of runs of the evolutionary optimization routine making this algorithm extremely computationally demanding. As only the best parameter sets from each run are assumed to contribute into the final solution, this algorithm appears far from being optimized. The goal of this study is to modify the optimization routine in a way that 20-40 runs would be enough to obtain qualitatively the same characterization. However, to keep the solution diversity throughout the HEO run, fitness sharing and newly developed shaking mechanisms are applied and tested on various test EPR spectra. In addition, other evolutionary optimization parameters such as population size and probability of genetic operators were also varied to tune the algorithm. According to the testing examples a speed-up factor of 5-7 was achieved. PMID- 16309266 TI - Resolved fluorescence emission spectra of PRODAN in ethanol/buffer solvents. AB - The fluorescence steady-state emission spectra of lipophilic fluorescence probe PRODAN in ethanol/buffer solvents of different concentrations (0.3, 0.9, 3 mol L( 1) ethanol) were extensively studied and analytically described. The complex experimental spectra, corrected for background effects, were fitted by two Gaussian curves. The energy separation of two maxima, (0.147+/-0.002) eV at 37 degrees C and (0.143+/-0.003) eV at 25 degrees C, was independent of ethanol concentration. The blue shifts observed for both maxima were linearly dependent on solvent polarity. The linear dependences of fluorescence's intensities on PRODAN concentration in all ethanol/buffer solvents indicate that no PRODAN self quenching takes place even at the highest measured PRODAN concentrations. PMID- 16309267 TI - Effect of lysine-28 side-chain acetylation on the nanomechanical behavior of alzheimer amyloid beta25-35 fibrils. AB - Amyloid fibrils are self-associating filamentous structures formed from the 39- to 42-residue-long amyloid beta peptide (Abeta peptide). The deposition of Abeta fibrils is one of the most important factors in the pathogenesis of Alzheimer's disease. Abeta25-35 is a fibril-forming peptide that is thought to represent the biologically active, toxic form of the full-length Abeta peptide. We have recently shown that beta sheets can be mechanically unzipped from the fibril surface with constant forces in a reversible transition, and the unzipping forces differ in fibrils composed of different peptides. In the present work, we explored the effect of epsilon-amino acetylation of the Lys28 residue on the magnitude of the unzipping force of Abeta25-35 fibrils. Although the gross structure of the Lys28-acetylated (Abeta25-35_K28Ac) and wild-type Abeta25-35 (Abeta25-35wt) fibrils were similar, as revealed by atomic force microscopy, the fundamental unzipping forces were significantly lower for Abeta25-35_K28Ac (20 +/ 4 pN SD) than for Abeta25-35wt (42 +/- 9 pN SD). Simulations based on a simple two-state model suggest that the decreased unzipping forces, caused most likely by steric constraints, are likely due to a destabilized zippered state of the fibril. PMID- 16309268 TI - Receptor occupancy on an ellipsoidal cell in the presence of a point source of a chemoattractant. AB - Diffusion of a chemoattractant from a micropipet is routinely used to examine the different aspects of a cell's chemotactic response. To quantify the effect of cell elongation on chemotactic sensitivity in the micropipet assay, the chemoattractant concentration at the cell plasma membrane was determined by solving the equation for diffusion from a point source in the presence of a prolate ellipsoid of varying eccentricity. The results show that cell elongation can significantly increase the difference in receptor occupancy between near and far cell ends and thereby enhance the sensitivity of chemotactic cells to shallow chemoattractant gradients. PMID- 16309269 TI - The influence of anisotropic membrane inclusions on curvature elastic properties of lipid membranes. AB - A membrane inclusion can be defined as a complex of protein or peptide and the surrounding significantly distorted lipids. We suggest a theoretical model that allows for the estimation of the influence of membrane inclusions on the curvature elastic properties of lipid membranes. Our treatment includes anisotropic inclusions whose energetics depends on their in-plane orientation within the membrane. On the basis of continuum elasticity theory, we calculate the inclusion-membrane interaction energy that reflects the protein or peptide induced short-ranged elastic deformation of a bent lipid layer. A numerical estimate of the corresponding interaction constants indicates the ability of inclusions to sense membrane bending and to accumulate at regions of favorable curvature, matching the effective shape of the inclusions. Strongly anisotropic inclusions interact favorably with lipid layers that adopt saddlelike curvature; such structures may be stabilized energetically. We explore this possibility for the case of vesicle budding where we consider a shape sequence of closed, axisymmetric vesicles that form a (saddle-curvature adopting) membrane neck. It appears that not only isotropic but also strongly anisotropic inclusions can significantly contribute to the budding energetics, a finding that we discuss in terms of recent experiments. PMID- 16309270 TI - Determination of partition coefficient of spin probe between different lipid membrane phases. AB - Model lipid membranes made from binary mixtures of dimyristoylphosphatidylcholine/dipalmitoylphosphatidylcholine (DMPC/DPPC) and dimyristoylphosphatidylcholine/cholesterol (DMPC/Chol) exhibit coexistence of diverse lipid phases at appropriate temperature and composition. Since lipids in different phases show different structural and motional properties, it is expected that the corresponding spin probe electron paramagnetic resonance (EPR) spectra will be superposition of several spectral components. From comparison of proportions of spectral components of the EPR spectrum with the fractions of the corresponding lipid phases obtained from known phase diagrams the partition coefficient of spin probe methyl ester of 5-doxyl palmitate between different lipid phases was determined. The results indicate that the used spin probe partitions approximately equally between different phases. PMID- 16309271 TI - Influence of fluctuations in actin structure on myosin V step size. AB - We study the influence of helical disorder in the actin structure on the myosin V step size, predicted from the elastic lever arm model. We show that fluctuations of +/-5 degrees per actin subunit, as proposed by Egelman et al., significantly alter the distribution of step sizes and improve the agreement with experimental data. PMID- 16309272 TI - Shape and size of giant unilamellar phospholipid vesicles containing cardiolipin. AB - The effect of cardiolipin content on the shape and size of giant palmitoyloleylphosphatidylcholine/cardiolipin vesicles was studied. Unilamellar vesicles were prepared in sugar solution by the method of electroformation, from mixtures containing up to 50% weight ratio of cardiolipin. At room temperature the vesicles containing cardiolipin exhibited abrupt changes in the curvature of the vesicle contour indicating regions of phase separation. The deviations from the spherical shape were larger if vesicles were made from mixtures with a higher content of cardiolipin. Numerous vesicles with soft fluctuating walls were observed. The estimated size of the vesicles containing cardiolipin was found to be smaller than the size of pure palmitoyloleylphosphatidylcholine vesicles. PMID- 16309273 TI - Effects of 5-fluorouracil on erythrocytes in relation to its cardiotoxicity: membrane structure and functioning. AB - In the present study, we showed that the antineoplastic drug 5-fluorouracil (5 FU) induces in vitro exposure-time/dose-dependent changes on the level of an erythrocyte's morphology, ionic balance, and membrane fluidity. These changes are partially or fully irreversible, and we suggest that they are provoked by an irreversible depletion of ATP. Because of these changes that could also occur in vivo during 5-FU infusion, a certain amount of erythrocytes with echinocytic shape and diminished ability to deliver oxygen is present in blood for longer periods of time. This renders oxygen transport and delivery more difficult, leaving the heart with an insufficient supply of oxygen, thus leading to cardiotoxicity. PMID- 16309274 TI - Blood clot dissolution dynamics simulation during thrombolytic therapy. AB - Nonocclusive blood clots only partially fill blood vessels and together with the adjacent vessel wall form a channel through which blood flows at usually much higher velocities than in normal vessels. Our aim was to find a theoretical explanation for the experimentally observed fact that fast flowing blood through the channel has a large effect on the increase of the clot dissolution rate compared to the dissolution rate in the absence of flow. Blood flow through the channel increases transport of dissolution agents to the clot and also exerts large forces to the surface of the clot along the channel. Proposed is a model for clot dissolution which assumes that the clot dissolution rate is proportional to the forces of flowing blood to the surface of the clot multiplied by the average blood velocity. The model has been verified by fitting to experimental magnetic resonance imaging data obtained by dynamical magnetic resonance microscopy of clots dissolved by recombinant tissue plasminogen activator in an artificial blood flow system. PMID- 16309275 TI - Study of the effect of ion channel modulators on photosynthetic oxygen evolution. AB - Various ion channel activities can be recorded by electrophysiological methods in the outer and inner envelope membranes of chloroplasts as well as in the thylakoid membrane. However, most of these channels are poorly characterized from a pharmacological point of view. Furthermore, the molecular identity has been determined only for a few of them, preventing an understanding of their role in plant physiology. By allowing specific ion fluxes across plastidial membranes, these ion channels may either directly or indirectly regulate photosynthesis, as has been hypothesized earlier. We have determined the effect of various ion channel modulators [indole-3-acetic acid, 5-nitro-2-(3-phenylpropylamino) benzoate, (-)-epigallocatechin-3-gallate, p-chlorophenoxyacetic acid, Konig's polyanion, Cs+, Gd3+, 4-aminopyridine, tetraethylammonium chloride, charybdotoxin, nimodipine, and cyclosporin A] on the efficiency of photosynthetic oxygen evolution in intact chloroplasts, broken chloroplasts, and isolated thylakoids. The data may improve our understanding of chloroplast ion channels and identifies inhibitors which may be exploited for electrophysiological studies. PMID- 16309276 TI - Membrane switch hypothesis. 1. Cell density influences lateral domain structure of tumor cell membranes. AB - The domain structure of human cancer cells membranes was investigated by electron paramagnetic resonance (EPR) in different phases of cell growth, and the results were compared to those obtained for nonmalignant cells. On the basis of computer simulation of the EPR spectra using a newly developed GHOST condensation routine it was suggested that plasma membranes of cancer cells have less lateral lipid domain types at confluent conditions than in the exponential growing phase, while in nonmalignant cells the domain structure does not change significantly during cell growth. In accordance to our experimental data we propose a membrane switch hypothesis: disappearance of certain membrane domain types might act as a switch promoting the clustering of membrane constituents into the active units in a common lipid membrane domain and thus influencing the physiology of cells. PMID- 16309277 TI - Membrane switch hypothesis. 2. Domain structure of phagocytes in horses with recurrent airway obstruction. AB - The mechanism of recurrent airway obstruction (RAO) in horses was investigated by measuring the membrane domain structure and oxy-redoxy activity in phagocytes isolated from bronchoalveolar lavage fluid (BAL) and from the blood of healthy and RAO horses by electron paramagnetic resonance (EPR). Differences in the activity of intracellular antioxidant enzymes CAT, GPx, and SOD measured in phagocytes of RAO horses in comparison to healthy horses showed that the phagocytes were affected by oxidative stress. In comparison with polymorphonuclear leukocytes (phagocytes) from the blood of healthy horses the reduction mechanisms in BAL were faster and coincided with the merging of disordered membrane domains, while in horses with RAO the reduction and membrane domain structure remained unchanged. We assume that the merging of lipid domains observed in phagocytes from BAL of healthy horses could promote cluster formation of membrane proteins or ligands, which could trigger the activation process in phagocytes of healthy horses and consequently the physiological response that probably did not happen in phagocytes of RAO horses. PMID- 16309278 TI - Detection of hydrogen atom adduct of spin-trap DEPMPO. The relevance for studies of biological systems. AB - We proposed EPR spectroscopy using spin-trap DEPMPO as a novel method for the detection of a hydrogen atom (*H) produced by chemical and biological systems. In complex EPR spectra of DEPMPO adducts in biological systems, spectral lines of unknown origin have been observed. We have assumed (Bacic, G.; Mojovic, M. Ann. N. Y. Acad. Sci. 2005, 1048, 230-243) that those lines represent the spectrum of a hydrogen atom (*H) adduct i.e., DEPMPO/H. An electrochemical system known to produce only *H radicals was used here in order to obtain a separate spectrum of the DEPMPO/H adduct. An acquired spectrum as well as a computer spectral simulation of the DEPMPO/H adduct showed considerable resemblance with additional lines in the EPR spectra of DEPMPO adducts in biological systems-plant plasma membranes and cell walls. This shows that such a radical is produced by plants as well as that DEPMPO is suitable for detection in both electrochemical and biological systems. PMID- 16309279 TI - Topology-aided molecular design: the platonic molecules of genera 0 to 3. PMID- 16309280 TI - Factors affecting d-block metal-ligand bond lengths: toward an automated library of molecular geometry for metal complexes. AB - Metal-ligand (M-L) bond lengths for a range of ligands (carboxylates, chlorides, pyridines, water, tertiary phosphines, and alkenes) and a variety of metals have been retrieved from the Cambridge Structural Database, CSD. Analysis of the factors which affect M-L bond lengths (for example, ligand coordination mode, oxidation state, metal coordination number and geometry, spin and Jahn-Teller effects, and ligand trans to M-L bond) shows that it is generally possible to subdivide the M-L data sets systematically to obtain better defined, unimodal, bond length distributions with means and sample standard deviations (SSDs) which reflect the nature of the bond in question. Typically, the SSDs for the M-L data sets can be reduced to 0.04-0.05 A by these methods. This work is an extension to tables of bond lengths in organometallic compounds and coordination complexes published in 1989. The importance of the factors which affect M-L bond lengths for particular metal-ligand groups are discussed. From the case studies reported, an algorithm is proposed by which compilation of a library of molecular geometry for metal complexes may be automated. The points that need to be considered to produce such a molecular library from the data stored in the CSD are discussed. The development of such a library would allow users to retrieve chemically well defined geometric data rapidly and accurately. This should be of use, for example, to crystallographers and molecular modelers. PMID- 16309281 TI - Supervised self-organizing maps in drug discovery. 1. Robust behavior with overdetermined data sets. AB - The utility of the supervised Kohonen self-organizing map was assessed and compared to several statistical methods used in QSAR analysis. The self organizing map (SOM) describes a family of nonlinear, topology preserving mapping methods with attributes of both vector quantization and clustering that provides visualization options unavailable with other nonlinear methods. In contrast to most chemometric methods, the supervised SOM (sSOM) is shown to be relatively insensitive to noise and feature redundancy. Additionally, sSOMs can make use of descriptors having only nominal linear correlation with the target property. Results herein are contrasted to partial least squares, stepwise multiple linear regression, the genetic functional algorithm, and genetic partial least squares, collectively referred to throughout as the "standard methods". The k-nearest neighbor (kNN) classification method was also performed to provide a direct comparison with a different classification method. The widely studied dihydrofolate reductase (DHFR) inhibition data set of Hansch and Silipo is used to evaluate the ability of sSOMs to classify unknowns as a function of increasing class resolution. The contribution of the sSOM neighborhood kernel to its predictive ability is assessed in two experiments: (1) training with the k-means clustering limit, where the neighborhood radius is zero throughout the training regimen, and (2) training the sSOM until the neighborhood radius is reduced to zero. Results demonstrate that sSOMs provide more accurate predictions than standard linear QSAR methods. PMID- 16309282 TI - Lead validation and SAR development via chemical similarity searching; application to compounds targeting the pY+3 site of the SH2 domain of p56lck. AB - Compound selection based on chemical similarity has been used to validate active "parent" compounds identified via database searching as viable lead compounds and to obtain initial structure-activity relationships for those leads. Twelve parent compounds that have inhibitory activity against the SH2 domain of the p56 T-cell tyrosine kinase (Lck) are the focus of this study. Lck is involved in the T-cell mediated immune response, and inhibitors of Lck protein-protein interactions could potentially be used to develop novel immunosuppressants. Similarity searches for each parent compound were performed using 2D structural fingerprints on a database containing 1,300,000 commercially available compounds. The inhibitory activity of the selected compounds was assessed using enzyme immunoassay (EIA). In general, the most active parent compounds yield the most high activity similar compounds; however, in two cases low activity parent compounds (i.e. inhibitory activity < 25% at 100 microM) yielded multiple similar compounds with activities > 60%. Such compounds may, therefore, be considered as viable lead compounds for optimization. Structure-activity relationships were explored by examining both ligand structures and their computed bound conformations to the protein. Functional groups common to the active compounds as well as key amino acid residues that form hydrogen bonds with the active compounds were identified. This information will act as the basis for the rational optimization of the lead compounds. PMID- 16309283 TI - QSAR model for predicting pesticide aquatic toxicity. AB - A hierarchical QSAR approach was applied for the prediction of acute aquatic toxicity. Chemical structures were encoded into molecular descriptors by an automated, seamless procedure available within the OpenMolGRID system. Finally, various linear and nonlinear regression techniques were used to obtain stable and thoroughly validated QSARs. The final model was developed by a counterpropagation neural network coupled with genetic algorithms for variable selection. The proposed QSAR is consistent with McFarland's principle for biological activity and makes use of seven molecular descriptors, namely HACA-2, HOMO-LUMO energy gap, Kier and Hall index, HA dependent HDSA-1, BETA polarizability, FHBCA fractional HBSA, and LogP. The model was extensively tested by the test set (R2= 0.79), the y-scrambling test, and sensitivity/stability tests. PMID- 16309284 TI - Structure-based classification of chemical reactions without assignment of reaction centers. AB - The automatic classification of chemical reactions is of high importance for the analysis of reaction databases, reaction retrieval, reaction prediction, or synthesis planning. In this work, the classification of photochemical reactions was investigated with no explicit assignment of the reacting centers. Classifications were explored with Random Forests or Kohonen neural networks in three different situations, using different levels of information: (a) pairs of reactants were classified according to the type of reaction they produce, (b) products were classified according to the type of reaction from which they can be synthesized, and (c) reactions were classified from the difference between the descriptors of the product and the descriptors of the reactants. In all cases molecular maps of atom-level properties (MOLMAPs) were used as descriptors. They are generated by a self-organizing map and encode physicochemical properties of the bonds available in a molecule. Correct classification could be achieved for approximately 90% of the 78 reactions in an independent test set. PMID- 16309285 TI - Novel statistical approach for primary high-throughput screening hit selection. AB - The standard activity threshold-based method (the "top X" approach), currently widely used in the high-throughput screening (HTS) data analysis, is ineffective at identifying good-quality hits. We have proposed a novel knowledge-based statistical approach, driven by the hidden structure-activity relationship (SAR) within a screening library, for primary hit selection. Application to an in-house ultrahigh-throughput screening (uHTS) campaign has demonstrated it can directly identify active scaffolds containing valuable SAR information with a greatly improved confirmation rate compared to the standard "top X" method (from 55% to 85%). This approach may help produce high-quality leads and expedite the hit-to lead process in drug discovery. PMID- 16309286 TI - Prediction of properties from simulations: a re-examination with modern statistical methods. AB - We discuss models fit to data collected by Duffy and Jorgensen to predict solvation free energies and partition equilibria of drugs, organic molecules, aromatic heterocycles, and other molecules. These data were originally examined using linear regression, but here more recently developed statistical models are applied. The data set is complicated due to the presence of discrepant observations and also curvature in the response. In some cases it is possible to discard a small number of the observations to get good fit to the data, but, in others, discarding an increasing proportion of the observations does not improve the fit. Our general preference is to use robust parameter estimation which downweights to reduce the influence of discrepant observations on the fitted models. Models are selected for four responses using linear or more complicated representations of the explanatory variables, such as cubic polynomials, B splines, or smoothers via generalized additive models (GAMs). Variables are chosen using the traditional approach of formal tests to assess contribution to the fit of a model, and resampling methods including bootstrap are also considered to assess the prediction error for given models. Results of our analysis indicate that GAMs are an improvement on linear models for describing the data and making predictions. In general robust regression models and GAMs have the smallest conditional expected loss of prediction over the four responses. In addition, robust regression models offer the advantage of identifying molecules that perform poorly in the fit. In general, models were identified that yielded an improvement of approximately 50% in the conditional expected loss of prediction compared with the original parametrization of Duffy and Jorgensen. It was also found that the use of cross-validation to compare models was unreliable, and bootstrapping is preferred. PMID- 16309287 TI - Studying the explanatory capacity of artificial neural networks for understanding environmental chemical quantitative structure-activity relationship models. AB - Although artificial neural networks (ANNs) have been shown to exhibit superior predictive power in the study of quantitative structure-activity relationships (QSARs), they have also been labeled a "black box" because they provide little explanatory insight into the relative influence of the independent variables in the predictive process so that little information on how and why compounds work can be obtained. Here, we have turned our interests to their explanatory capacities; therefore, a method was proposed for assessing the relative importance of variables indicating molecular structure, on the basis of axon connection weights and partial derivatives of the ANN output with respect to its input, which can identify variables that significantly contribute to network predictions, and providing a variable selection method for ANNs. We show that, by extending this approach to ANNs, the "black box" mechanics of ANNs can be greatly illuminated, thereby making it very useful in understanding environmental chemical QSAR models. PMID- 16309288 TI - Anatomy of fingerprint search calculations on structurally diverse sets of active compounds. AB - Similarity searching using molecular fingerprints is a widely used approach for the identification of novel hits. A fingerprint search involves many pairwise comparisons of bit string representations of known active molecules with those precomputed for database compounds. Bit string overlap, as evaluated by various similarity metrics, is used as a measure of molecular similarity. Results of a number of studies focusing on fingerprints suggest that it is difficult, if not impossible, to develop generally applicable search parameters and strategies, irrespective of the compound classes under investigation. Rather, more or less, each individual search problem requires an adjustment of calculation conditions. Thus, there is a need for diagnostic tools to analyze fingerprint-based similarity searching. We report an analysis of fingerprint search calculations on different sets of structurally diverse active compounds. Calculations on five biological activity classes were carried out with two fingerprints in two compound source databases, and the results were analyzed in histograms. Tanimoto coefficient (Tc) value ranges where active compounds were detected were compared to the distribution of Tc values in the database. The analysis revealed that compound class-specific effects strongly influenced the outcome of these fingerprint calculations. Among the five diverse compound sets studied, very different search results were obtained. The analysis described here can be applied to determine Tc intervals where scaffold hopping occurs. It can also be used to benchmark fingerprint calculations or estimate their probability of success. PMID- 16309289 TI - Virtual hydrocarbon and combinatorial databases for use with CAVEAT. AB - Three new virtual databases have been developed for use with the bond-orientation based database searching program CAVEAT. These consist of a database of trisubstituted monocyclic hydrocarbons having ethyl, vinyl, and phenyl substituents; a database of unsubstituted bicyclic hydrocarbons; and a database of core structures from established combinatorial synthetic methods having hydrogen, ethyl, vinyl, and phenyl substituents at the readily varied positions. Each collection of molecules was subjected to a batch conformational search, minimization, and conversion to a vector database for use with CAVEAT. PMID- 16309290 TI - Identifying biologically active compound classes using phenotypic screening data and sampling statistics. AB - Scoring the activity of compounds in phenotypic high-throughput assays presents a unique challenge because of the limited resolution and inherent measurement error of these assays. Techniques that leverage the structural similarity of compounds within an assay can be used to improve the hit-recovery rate from screening data. A technique is presented that uses clustering and sampling statistics to predict likely compound activity by scoring entire structural classes. A set of phenotypic assays performed against a commercially available compound library was used as a test set. Using the class-scoring technique, the resultant activity prediction scores were more reproducible than individual assay measurements, and class scoring recovered known active compounds more efficiently than individual assay measurements because class scoring had fewer false positives. Known biologically active compounds were recovered 87% of the time using class scores, suggesting a low false-negative rate that compared well to individual assay measurements. In addition, many weak and potentially novel classes of active compounds, overlooked by individual assay measurements, were suggested. PMID- 16309291 TI - Theoretical study of changes in pi-electron delocalization in the analogues of an ortho-hydroxy schiff base when the proton is replaced with Li+ or BeH+. AB - Molecular geometries of ortho-hydroxy Schiff base in keto-enamine and enol-imine tautomeric forms, its anion, and their derivatives in which H+ was replaced with Li+ or BeH+ were optimized at the B3LYP/6-311+G level of theory. Isodesmic reactions for estimating delocalization due to H-bonding or cation chelating were calculated. Geometry-based aromaticity index HOMA and magnetism-based NICS1(zz) index were used to estimate pi-electron delocalization. Keto-enamine tautomer exhibits low aromaticity in the ring and a relatively high pi-electron delocalization in the quasi-ring. The reverse was found for enol-imine tautomer. The Li+ and BeH+ derivatives showed a relatively high pi-electron delocalization in the ring and in the quasi-ring. This may be interpreted by an extension of the electron delocalization path in the pi-electron system through low-lying unoccupied p-type orbitals of Li+ and BeH+ cations. PMID- 16309292 TI - 'Inductive' charges on atoms in proteins: comparative docking with the extended steroid benchmark set and discovery of a novel SHBG ligand. AB - We have developed a novel iterative approach for calculation of partial charges in proteins within the framework of the 'molecular capacitance' model. The method operates by an effective 'inductive' electronegativity scale derived from a number of the conventional charge systems including CHARMM, AMBER, MMFF, OPLS, and PEOE among others. Our novel 'inductive' electronegativity equalization procedure allows rapid and conformation sensitive computation of adequate partial charges in proteins. Accuracy of the 'inductive' values was confirmed by their correlation with DFT-computed partial charges in common amino acids. A comparative docking study with an extended steroid data set not only illustrated the adequacy of 'inductive' protein charges but also demonstrated their superior performance compared to several conventional protein charging systems. Subsequent docking with 'inductive' charges resulted in identification of five potential leads as human Sex Hormone Binding Globulin (SHBG) ligands from a commercial library of natural compounds. When the selected substances were evaluated for their ability to bind SHBG in vitro, three of them displaced testosterone from the SHBG steroid-binding site, and with one compound this was achieved at micromolar concentrations. PMID- 16309293 TI - Investigation of DNA-binding properties of an aminoglycoside-polyamine library using quantitative structure-activity relationship (QSAR) models. AB - We have recently developed a novel multivalent cationic library based on the derivatization of aminoglycosides by linear polyamines. In the current study, we describe the DNA-binding activity of this library. Screening results indicated that several candidates from the library showed high DNA-binding activities with some approaching those of cationic polymers. Quantitative Structure-Activity Relationship (QSAR) models of the screening data were employed to investigate the physicochemical effects governing polyamine-DNA binding. The utility of these models for the a priori prediction of polyamine-DNA-binding affinity was also demonstrated. Molecular descriptors selected in the QSAR modeling indicated that molecular size, basicity, methylene group spacing between amine centers, and hydrogen-bond donor groups of the polyamine ligands were important contributors to their DNA-binding efficacy. The research described in this paper has led to the development of new multivalent ligands with high DNA-binding activity and improved our understanding of structure-activity relationships involved in polyamine-DNA binding. These results have implications for the discovery of novel polyamine ligands for nonviral gene delivery, plasmid DNA purification, and anticancer therapeutics. PMID- 16309294 TI - Prediction of the rodent carcinogenicity of 60 pesticides by the DEREKfW expert system. AB - The two-year rodent bioassay represents the golden standard for evaluating the carcinogenicity of chemicals. Because of practical and ethical reasons, alternative approaches have been investigated for many years. Among these approaches, the (quantitative) structure-activity relationships [(Q)SARs] offer promising perspectives for quickly screening a large number of chemicals. To increase the acceptance of (Q)SARs among the regulators, their predictive power needs to be scientifically validated. In this article, we tested the capacity of the DEREKfW expert system to qualitatively predict the rodent carcinogenicity and the genotoxic potential of 60 pesticides recently registered in Switzerland. The percentage of false negatives was found to be 31% for carcinogenicity. The associated sensitivity of 69% indicates that most of the pesticides with positive rodent bioassay results were detected by DEREKfW. On the other hand, the low specificity of 47% indicates that many pesticides may be flagged as carcinogenic while rodent bioassays would not confirm this potential. This may lead to unnecessary testing or the unnecessary restriction of a chemical. PMID- 16309295 TI - Comparing the performance of FLUFF-BALL to SEAL-CoMFA with a large diverse estrogen data set: from relevant superpositions to solid predictions. AB - In this work a template-based molecular mechanistic superposition algorithm FLUFF (Flexible Ligand Unified Force Field) and an accompanying local coordinate QSAR method BALL (Boundless Adaptive Localized Ligand) are validated against the benchmark techniques SEAL (Steric and Electrostatic Alignment) and CoMFA (Comparative Molecular Field Analysis) using a large diverse set of 245 xenoestrogens extracted from the EDKB (Endocrine Disruptor Knowledge Base) maintained by NCTR (National Centre for Toxicological Research). The results indicate that FLUFF is capable of generating relevant superpositions not only for BALL but also for CoMFA, as both techniques give predictive QSAR models. When the BALL and CoMFA methods are compared, it is clear that the BALL algorithm met or even exceeded the results of the standard 3D-QSAR method CoMFA using alignments either from the tailor-made superposition technique FLUFF or the reference method SEAL. The FLUFF-BALL method can be easily automated, and it is computationally light, providing thus a good computational "sieve" capable of fast screening of large molecule libraries. PMID- 16309296 TI - Modeling of cyclin-dependent kinase inhibition by 1H-pyrazolo[3,4-d]pyrimidine derivatives using artificial neural network ensembles. AB - Artificial neural network ensembles were used for modeling the cyclin-dependent kinase inhibition of 1H-pyrazolo[3,4-d]pyrimidine derivatives. The structural characteristics of these inhibitors were encoded in relevant 3D-spatial descriptors extracted by genetic algorithm feature selection. Bayesian regularized multilayer neural networks, trained by the back-propagation algorithm, were developed using these variables as inputs. The predictive power of the model was tested by leave-one-out cross validation. In addition, for a more rigorous measure of the predictive capacity, multiple validation sets were randomly generated as members of neural network ensembles, which makes doing averaged predictions feasible. In this way, the predictive power was analyzed accounting for the averaged test set R values and test set mean-square errors. Otherwise, Kohonen self-organizing maps were used as an additional tool for the same modeling. The location of the inhibitors in a map facilitates the analysis of the connection between compounds and serves as a useful tool for qualitative predictions. PMID- 16309297 TI - In silico footprinting of ligands binding to the minor groove of DNA. AB - The sequence selectivity of small molecules binding to the minor groove of DNA can be predicted by "in silico footprinting". Any potential ligand can be docked in the minor groove and then moved along it using simple simulation techniques. By applying a simple scoring function to the trajectory after energy minimization, the preferred binding site can be identified. We show application to all known noncovalent binding modes, namely 1:1 ligand:DNA binding (including hairpin ligands) and 2:1 side-by-side binding, with various DNA base pair sequences and show excellent agreement with experimental results from X-ray crystallography, NMR, and gel-based footprinting. PMID- 16309298 TI - Considerations in compound database preparation--"hidden" impact on virtual screening results. AB - Structure-based virtual screening (SBVS) utilizing docking algorithms has become an essential tool in the drug discovery process, and significant progress has been made in successfully applying the technique to a wide range of receptor targets. In silico validation of virtual screening protocols before application to a receptor target using a corporate or commercially available compound collection is key to establishing a successful process. Ultimately, retrieval of a set of active compounds from a database of inactives is required, and the metric of enrichment (E) is habitually used to discern the quality of separation of the two. Numerous reports have addressed the performance of docking algorithms with regard to the quality of binding mode prediction and the issue of postprocessing "hit lists" of docked ligands. However, the impact of ligand database preprocessing has yet to be examined in the context of virtual screening and prioritization of compounds for biological evaluation. We provide an insight into the implications of cheminformatic preprocessing of a validation database of compounds where multiple protonated, tautomeric, stereochemical, and conformational states have been enumerated. Several commonly used methods for the generation of ligand conformations and conformational ensembles are examined, paired with an exhaustive rigid-body algorithm for the docking of different "multimeric" compound representations to the ligand binding site of the human estrogen receptor alpha. Chemgauss, a shapegaussian scoring function with intrinsic chemical knowledge, was combined with PLP as a consensus-scoring scheme to rank output from the docking protocol and enrichment rates calculated for each screen. The overheads of CPU consumption and the effect on relative database size (disk requirement) for each of the protocols employed are considered. Assessment of these parameters indicates that SBVS enrichments are highly dependent on the initial cheminformatic treatment(s) used in database construction. The interplay of SMILES representations, stereochemical information, protonation state enumeration, and ligand conformation ensembles are critical in achieving optimum enrichment rates in such screening. PMID- 16309299 TI - Improving the quality of 3D-QSAR by using flexible-ligand receptor models. AB - To address the problems associated with molecular conformations and alignments in the 3D-QSAR studies, we have developed the Flexible Ligand - Atomic Receptor Model (FLARM) 2.0 method. The FLARM 2.0 method has three unique features as compared to other pseudoreceptor model methods: (1) the training ligands are flexibly optimized inside the receptors to achieve minimal docking energies; (2) the receptor atoms are spatially moveable in the process of genetic evolving in order to avoid improper initial receptor shapes; and (3) void receptor sites are specially favored in order to obtain open receptor models that allow large gaps. Advantages of an open model include less noise information, a smaller risk of overfitting, and ease of locating the key interaction sites. The latter two features, inherited from the previous FLARM 1.0 method, can improve the predictive ability of the 3D-QSAR models, while the first feature is newly implemented to relieve the uncertainty caused by improper conformation and alignment. Three FLARM 2.0 case studies were performed, and the results show that FLARM 2.0 models are highly predictive and robust. FLARM 2.0 pseudoreceptor models can correspond well with the pharmacophore models and/or the binding sites of the real protein receptors. PMID- 16309300 TI - Ligand-based molecular modeling study on a chemically diverse series of cholecystokinin-B/gastrin receptor antagonists: generation of predictive model. AB - Pharmacophore hypotheses were developed for six structurally diverse series of cholecystokinin-B/gastrin receptor (CCK-BR) antagonists. A training set consisting of 33 compounds was carefully selected. The activity spread of the training set molecules was from 0.1 to 2100 nM. The most predictive pharmacophore model (hypothesis 1), consisting of four features, namely, two hydrogen bond donors, one hydrophobic aliphatic, and one hydrophobic aromatic feature, had a correlation (r) of 0.884 and a root-mean-square deviation of 1.1526, and the cost difference between null cost and fixed cost was 81.5 bits. The model was validated on a test set consisting of six different series of 27 structurally diverse compounds and performed well in classifying active and inactive molecules correctly. This validation approach provides confidence in the utility of the predictive pharmacophore model developed in this work as a 3D query tool in the virtual screening of drug-like molecules to retrieve new chemical entities as potent CCK-BR antagonists. The model can also be used to predict the biological activities of compounds prior to their costly and time-consuming synthesis. PMID- 16309301 TI - Parallelizing a molecular dynamics algorithm on a multiprocessor workstation using OpenMP. AB - The atomistic molecular dynamics program YASP has been parallelized for shared memory computer architectures. Parallelization was restricted to the most CPU time-consuming parts: neighbor-list construction, calculation of nonbonded, angle and dihedral forces, and constraints. Most of the sequential FORTRAN code was kept; parallel constructs were inserted as compiler directives using the OpenMP standard. Only in the case of the neighbor list did the data structure have to be changed. The parallel code achieves a useful speedup over the sequential version for systems of several thousand atoms and above. On an IBM Regatta p690+, the throughput increases with the number of processors up to a maximum of 12-16 processors depending on the characteristics of the simulated systems. On dual processor Xeon systems, the speedup is about 1.7. PMID- 16309302 TI - McQSAR: a multiconformational quantitative structure-activity relationship engine driven by genetic algorithms. AB - The generation of quantitative structure-activity relationships (QSARs) under the supervision of a genetic algorithm (GA) is a QSAR modeling approach used for more than a decade. In this paper we present McQSAR, an extension to the traditional GA approach to derive QSARs. McQSAR is able to use descriptors for multiple representations per compound, such as different conformers, tautomers, or protonation forms. Test runs show that the algorithm converges to a set of representations that describe the binding mode of the set of input molecules to a reasonable resolution provided that suitable descriptors-based on the three dimensional structure-are used. Furthermore, the frequency of chance correlation was measured during multiple runs on a real-life data set using simulated linear relationship functions. The observed frequency of chance correlation, on average 0.3 +/- 0.5%, was found independent of the size of the calibration set and the number of terms in the underlying relationship function. PMID- 16309303 TI - Extraction, hemisynthesis, and synthesis of canthin-6-one analogues. Evaluation of their antifungal activities. AB - Zanthoxylum chiloperone var. angustifolium was investigated. Alkaloids 1-3 from the canthin-6-one series were characterized. Derivatives 7-28 were prepared by hemisynthesis or total synthesis. All compounds were tested for in vitro antifungal activities against five pathogenic fungal strains. Analogues of canthin-6-one did not show better antifungal activities. PMID- 16309304 TI - Leucovernine and acetylleucovernine, alkaloids from Leucojum vernum. AB - The fresh bulbs of Leucojum vernum provided seven tyrosine-derived alkaloids; two of them have not been reported before and are named leucovernine and acetylleucovernine. The five known alkaloids were N-demethylgalanthamine, hippeastrine, 9-O-demethylhomolycorine, 5alpha-hydroxyhomolycorine, and 11 hydroxyvittatine. These compounds have been isolated from this species for the first time. The structure determination was carried out by the combination of liquid-phase one- and two-dimensional NMR spectroscopy and high-resolution mass spectrometry. PMID- 16309305 TI - Using jasplakinolide to turn on pathways that enable the isolation of new chaetoglobosins from Phomospis asparagi. AB - The isolation and structure elucidation of three new secondary metabolites, chaetoglobosin-510 (1), -540 (2), and -542 (3), are described. These compounds were produced by cultures of the marine-derived fungus Phomopsis asparagi, challenged with the known F-actin inhibitor jasplakinolide. Chaetoglobosin-542 (3) displayed antimicrofilament activity and was cytotoxic toward murine colon and leukemia cancer cell lines. PMID- 16309306 TI - Verticillane derivatives from Bursera suntui and Bursera kerberi. AB - The stems of Bursera suntui afforded two new verticillane derivatives, (1S,3Z,7E,11S,12S)-(+)-verticilla-3,7-dien-12,20-diol (1) and (1S,3Z,7E,11S,12S) (+)-verticilla-3,7-dien-12,20-diol 20-acetate (2), together with (1S,3E,7E,11R) (+)-verticilla-3,7,12(18)-triene (3), (1R,3E,7E,11R,12Z)-(+)-verticilla-3,7,12 triene (4), (1R,7E,11Z)-(-)-verticilla-4(20),7,11-triene (5), and (1S,3E,7E,11S,12S)-(+)-verticilla-3,7-dien-12-ol (6). Compounds 3 and 4 are new enantiomerically pure natural products whose racemic mixtures, derived from synthetic approaches toward the taxane skeleton, were obtained previously. The stems of Bursera kerberi afforded the new (1S,3E,7E,11S,12R)-(+)-verticilla-3,7 dien-12-ol (7) together with 3-5. This is the first time that verticillane derivatives have been isolated from the genus Bursera. Their structures and stereochemistry were elucidated by 1D and 2D NMR data, including COSY, NOESY, HSQC, and HMBC experiments, while the absolute configuration was determined by comparison of the optical rotatory dispersion data with that of recently revised (1S,3E,7E,11S,12S)-(+)-verticilla-3,7-dien-12-ol (6), obtained from Sciadopitys verticillata, and those of (1R,3E,7E,11R,12R)-(-)-verticilla-3,7-dien-12-ol (8) and (1R,3E,7E,11R,12S)-(-)-verticilla-3,7-dien-12-ol (9), isolated from the liverwort Jackiella javanica. The conformational preferences of 1-7 were studied by molecular mechanics modeling employing the Monte Carlo protocol. PMID- 16309307 TI - Structure and absolute configuration of nyasol and hinokiresinol via synthesis and vibrational circular dichroism spectroscopy. AB - The absolute configuration of the norlignan (+)-nyasol was determined to be S by comparison of the experimental vibrational circular dichroism data with first principle calculations taking into account the eight lowest energy conformations. The established absolute configuration of (+)-nyasol enables establishment of the absolute configuration of (-)-hinokiresinol, which is concluded to be S. A total synthesis and resolution of hinokiresinol has been performed to resolve the conflicting reports of the coupling constant of the vinylic protons of the disubstituted double bond in this molecule. Racemic hinokiresinol was resolved. Both enantiomers possess the same antiplasmodial activity. PMID- 16309308 TI - Anti-inflammatory activity of verminoside from Kigelia africana and evaluation of cutaneous irritation in cell cultures and reconstituted human epidermis. AB - Kigelia africana is a plant used in Africa for anti-inflammatory, anti-microbial, and anti-skin-aging effects. Various papers have reported on the composition and biological activities of its CH2Cl2 extracts and dermal formulations. Chemical analysis of a polar extract of fruit from K. africana indicated the presence of verminoside (1), an iridoid, as a major constituent, and of a series of polyphenols such as verbascoside (2). In vitro assays showed that 1 had significant anti-inflammatory effects, inhibiting both iNOS expression and NO release in the LPS-induced J774.A1 macrophage cell line. Cytotoxicity and cutaneous irritation of the extract and of compounds 1 and 2 were investigated. The crude extract and 1 did not affect cell viability in vitro either in cells grown in monolayers (ML) or in the reconstituted human epidermis (RHE, 3D) model; neither caused release of pro-inflammatory mediators or histomorphological modification of RHE. PMID- 16309309 TI - Free-radical-scavenging and xanthine oxidase inhibitory constituents from Stereospermum personatum. AB - Bioassay-guided fractionation of different extracts of both stem and stem bark of Stereospermum personatum led to the isolation of free-radical-scavenging and xanthine oxidase inhibitory molecules along with three new anthraquinones, sterequinones F-H (1-3), a new naphthoquinone, sterequinone I (4), two new phenethyl esters, 2(4'-hydroxyphenyl)ethyl undecanoate (14) and 2(4' hydroxyphenyl)ethyl nonacosanoate (15), and a new 3,4,5-trimethoxycinnamyl ether, 2-methoxy-4-[3'-(3'',4'',5''trimethoxyphenyl)allyloxymethyl]phenol (16), together with known compounds. The antioxidant and xanthine oxidase inhibitory potentials of the isolated compounds are reported. PMID- 16309310 TI - The epimerization of sesamin and asarinin. AB - Sesamin (1) and asarinin (2) are two C-7' epimeric lignans. Molecular modeling by semiempirical methods indicated that 1 is more stable than 2 by about 2.5 kcal/mol. However, epimerization under acidic conditions led to a 44.8/55.2 equilibrium ratio of 1 and 2. Single-crystal X-ray diffraction analyses indicated that 1 was monoclinic with a = 10.0435(19) A, b = 6.9151(8) A, c = 11.8460(13) A, and 2 was triclinic with a = 5.595(5) A, b = 9.5910(18) A, c = 15.620(4) A. The unexpected equilibrium ratio of 1 and 2 indicated that structural changes are dependent on the conditions of the extraction processes. PMID- 16309311 TI - (+)-Myristinins A and D from Knema elegans, which inhibit DNA polymerase beta and cleave DNA. AB - A survey of crude plant extracts for DNA polymerase beta inhibitors resulted in the identification of a methyl ethyl ketone extract prepared from Knema elegans that strongly inhibited the enzyme. Subsequent bioassay-guided fractionation of the extract, using an assay to monitor the activity of DNA polymerase beta, led to the isolation of two potent inhibitors, (+)-myristinins A (1) and D (2), which are known flavans having unusual structures. (+)-Myristinins A and D exhibited IC50 values of 12 and 4.3 microM, respectively, as inhibitors of DNA polymerase beta in the presence of bovine serum albumin (BSA), and 2.7 and 1.2 microM in the absence of BSA. As such, they are the most potent DNA polymerase beta inhibitors reported to date. Compounds 1 and 2 potentiated the cytotoxicity of bleomycin toward cultured P388D1 cells, reducing the number of viable cells by at least 30% when employed at 9 microM concentration for 6 h in the presence of an otherwise nontoxic concentration of bleomycin (75 nM). Principles 1 and 2 also induced strong Cu2+-dependent DNA strand scission in a DNA cleavage assay. Accordingly, 1 and 2 exhibit two activities, namely, DNA polymerase beta inhibition and DNA damage. PMID- 16309312 TI - Synthesis of deserpidine from reserpine. AB - The Rauwolfia alkaloids reserpine (1) and deserpidine (2), two alkaloids from Rauwolfia species, have been widely used for their antihypertensive action. Deserpidine (2) is a compound with limited availability from natural sources, and its synthesis from 1 in six steps (41% overall yield) is reported here. PMID- 16309313 TI - Oxygenated pimarane diterpenes from Kaempferia marginata. AB - Six new diterpenes, (1R,2S,5S,7S,9R,10S,13R)-1,2,7-trihydroxypimara-8(14),15 diene (1), (1R,2S,5S,9S,10S,11R,13R)-1,2,11-trihydroxypimara-8(14),15-diene (2), (1S,5S,7R,9R,10S,11R,13R)-1,7,11-trihydroxypimara-8(14),15-diene (3), (1S,5S,9S,10S,11R,13R)-1,11-dihydroxypimara-8(14),15-diene (4), (5S, 6R,9S,10S,13R)-6-hydroxypimara-8(14),15-diene-1-one (5), and (1R,2S,5S,7S,9R,10S,13R)-1,2-dihydroxypimara-8(14),15-diene-7-one (6), along with four known diterpenes, have been isolated from the dichloromethane extract of whole plants of Kaempferia marginata. The structures were assigned by spectroscopic methods. The absolute configuration of 1 was established by the Mosher ester method. Substances obtained were evaluated against a panel of bioassays including antimalarial, antituberculous, and antifungal activity. PMID- 16309314 TI - Coumarins from the fruits of Seseli devenyense. AB - Eight new coumarins were isolated from the fruits of Seseli devenyense Simonkai. Their structures were established from NMR and mass data and their absolute configurations from chemical degradation correlation reactions. The new structures are the decanoic and dodecanoic esters of (+)-lomatin (3, 4), the decanoates of (+)-cis-khellactone at positions 4' (5) and 3' (6) as well as the 2'S epimer of 8-(2,3-dihydroxy-3-methylbutyl)-7-hydroxychromen-2-one (7) named devenyol, its two O-monoglucosides at positions 3' and 7 named devenyosides A (8) and B (9), and the corresponding 3'- and 7-O-diglucoside named devenyoside C (10). This plant is an interesting example of stereochemical diversity based on biodiversity given that other members of the Apiaceae family produce exclusively the 2'R epimers of compounds 7-9. PMID- 16309315 TI - Digitoxin inhibits the growth of cancer cell lines at concentrations commonly found in cardiac patients. AB - The cardiac glycosides digitoxin (1) and digoxin (3) have been used in cardiac diseases for many years. During this time several reports have suggested the possible use of digitalis in medical oncology. Several analogues of digitoxin (1) were evaluated for growth inhibition activity in three human cancer cell lines; this study showed that digitoxin (1) was the most active compound and revealed some structural features that may play a role in the growth inhibition activity of these drugs. The IC50 values for 1 (3-33 nM) were within or below the concentration range seen in the plasma of patients with cardiac disease receiving this glycoside (20-33 nM). A renal adenocarcinoma cancer cell line (TK-10) was hypersensitive to this drug, and digitoxin toxicity on these cells was mediated by apoptosis. In vitro experiments showed that 1 at 30 nM induced levels of DNA topoisomerase II cleavable complexes similar to etoposide, a topoisomerase II poison widely used in cancer chemotherapy. Using the individual cell assay TARDIS, cells exposed to 1 for 30 min showed low but statistically significant levels of DNA-topoisomerase II cleavable complexes; however these complexes disappeared after 24 h exposure. PMID- 16309316 TI - Iridoid glycosides and grayanane diterpenoids from the roots of Craibiodendron henryi. AB - Four new iridoid glycosides, 10-O-trans-p-coumaroylscandoside (1), 10-O-cis-p coumaroylscandoside (2), 10-O-trans-p-coumaroyldesacetyl asperulosidic acid (3), and 10-O-cis-p-coumaroyldesacetyl asperulosidic acid (4), and two new grayanane diterpenoids, 14beta-O-(2S,3S-nilyl)-2alpha,3beta,5beta,6beta,16alpha pentahydroxygrayanane (5) and 14beta-O-(2S,3S-nilyl) 2alpha,3beta,5beta,6beta,16alpha-pentahydroxygrayan-10(20)-ene (6), have been isolated from Craibiodendron henryi. The structures of these compounds were determined by chemical and spectroscopic methods including 1H-1H COSY, HMQC, HMBC, and NOESY experiments. Antioxidant activities and vasodilator effects of these compounds were assessed. PMID- 16309317 TI - Meroditerpenoids from a Formosan soft coral Nephthea chabrolii. AB - Eight new meroditerpenoid-related metabolites, including one naphthoquinone derivative, chabrolonaphthoquinone B (1), four tetraprenyltoluquinone-related compounds, chabrolobenzoquinones E-H (2-5), and three tetraprenyltoluquinol related metabolites, chabrolohydroxybenzoquinones E-G (6-8), were isolated from the organic extract of a Taiwanese soft coral Nephthea chabrolii. The structures of 1-8 were elucidated on the basis of extensive spectroscopic analysis and by comparison of the data with those of the related metabolites. Cytotoxic activity of metabolites 1-3 and 5-8 against a limited panel of cancer cell lines is also described. PMID- 16309318 TI - Bioactive tetrahydrofuran lignans from Peperomia dindygulensis. AB - Five new tetrahydrofuran lignans (1-5), accompanied by four known compounds, were isolated from the ethyl acetate extract of Peperomia dindygulensis. Structures were elucidated mainly using 1D NMR, 2D NMR, and mass spectroscopic studies. The relative configurations of 1-5 were determined by NOE correlations. Several of the compounds showed weak growth inhibitory activity against three cell lines (WI 38, VA-13, and HepG2). Compound 5 exhibited stronger MDR (multidrug resistance) reversal activity than verapamil at 2.5 microg/mL in a cellular calcein accumulation assay. Compounds 4 and 5 showed weak inhibitory activity against induction of the intercellular adhesion molecule-1 (ICAM-1) in anti-inflammatory activity experiments. PMID- 16309319 TI - Roquefortine E, a diketopiperazine from an Australian isolate of Gymnoascus reessii. AB - The new isoprenylated diketopiperazine roquefortine E (6) has been isolated from an Australian soil isolate of the ascomycete Gymnoascus reessii. The known fungal metabolite roquefortine C (1) was also recovered as the major antibacterial principle, and all structures were assigned by detailed spectroscopic analysis. PMID- 16309320 TI - Cytotoxic clerodane diterpenoids from Casearia membranacea. AB - Bioassay-guided fractionation of the EtOAc extract of Casearia membranacea leaves and twigs afforded three new clerodane diterpenes, caseamembrins M-O (1-3), and the known rel-(2S,5R,6R,8S,9S,10R,18S,19R)-2-(2-methylbutyryloxy)-6-hydroxy-18,19 di-O-acetyl-18,19-epoxycleroda-3,13(16),14-triene (4) and caseamembrin D (5). The structures of 1-3, including the relative configurations, were established by extensive NMR spectroscopic analyses. The cytotoxic activities of the isolated diterpenoids against human oral epidermoid (KB), medulla (Med), and colon (DLD-1) cancer cell lines were evaluated. PMID- 16309321 TI - Triterpenoid oligoglycosides from the sea cucumber Stichopus parvimensis. AB - Two new holostan-type triterpenoid oligoglycosides, named parvimosides A (1) and B (2), were isolated from the warty sea cucumber Stichopus parvimensis (syn. Parastichopus parvimensis). Their structures and total assignments of the proton and carbon signals were established on the basis of spectral evidence, mainly 1H and 13C 1D and 2D NMR spectroscopy including DEPT, COSY, TOCSY, NOESY/ROESY, HMQC/HSQC, and HMBC experiments, as well as FABMS. PMID- 16309322 TI - Hirsutane sesquiterpenes from the fungus Lentinus connatus BCC 8996. AB - Two new hirsutane sesquiterpenes, connatusins A (1) and B (2), were isolated from the fungus Lentinus connatus BCC 8996. The structures, closely related to hypnophilin, were elucidated on the basis of the spectroscopic data. An X-ray analysis was performed to confirm the structure of 1. Six known compounds were also obtained. Panepoxydone (5), panepoxydione (6), and dihydrohypnophilin (8) exhibited significant antimalarial and cytotoxic activities. PMID- 16309323 TI - Cytotoxic sesquiterpenes from Aplysia dactylomela. AB - Three new chamigrenes, compound 1, acetyldeschloroelatol 2 (2-bromo-1,1,9 trimethyl-5-methylenespiro[5.5]undec-8-en-3-yl acetate), and acetylelatol 4 (2 bromo-8-chloro-1,1,9-trimethyl-5-methylenespiro[5.5]undec-8-en-3-yl acetate), and the known metabolites deschloroelatol 3, elatol 5, 8-acetylcaespitol 6, and caespitol 7 have been isolated from the sea hare Aplysia dactylomela. The structures of 1, 2, and 4 were determined on the basis of spectroscopic evidences. The in vitro cytotoxicity of six of these compounds against two cancer cell lines (HeLa and Hep-2) and nontumoral Vero cells was evaluated. The results support the hypothesis that the acetate derivatives decrease the toxicity of the corresponding alcohols in a strategy to store toxic metabolites acquired through the diet. PMID- 16309324 TI - Cyclohexadepsipeptides from the insect pathogenic fungus Hirsutella nivea BCC 2594. AB - Two new cyclohexadepsipeptides, hirsutatins A (1) and B (2), were isolated from a culture filtrate of the insect pathogenic fungus Hirsutella nivea BCC 2594. Structures of these compounds were elucidated primarily by NMR and mass spectroscopic analyses. The alpha-carbon stereochemistry of 1 was established by HPLC analysis of its acid hydrolysate using a chiral column. Hirsutatin B (2) exhibited activity against the malarial parasite Plasmodium falciparum K1 with an IC50 value of 5.8 microg/mL, while hirsutatin A (1) was inactive at a concentration of 20 microg/mL. PMID- 16309325 TI - Immunosuppressive iridoids from the fruits of Gardenia jasminoides. AB - A new iridoid, gardaloside (1), and a new safranal-type monoterpene, jasminoside G (2), together with 10 known compounds including nine iridoids and a second safranal-type monoterpene, were isolated from the fruits of Gardenia jasminoides. The structures of 1 and 2 were established on the basis of spectroscopic evidence. Of these compounds, geniposide (3), 6alpha-hydroxygeniposide (5), ixoroside (7), and shanzhiside (8) showed significant inhibition of IL-2 secretion by phorbol myristate acetate and anti-CD28 monoclonal antibody co stimulated activation of human peripheral blood T cells. PMID- 16309326 TI - Dichotomins J and K, vasodilator cyclic peptides from Stellaria dichotoma. AB - Two new cyclic peptides, dichotomins J (1) and K (2), have been isolated from the roots of Stellaria dichotoma, and their structures were elucidated by chemical degradation and extensive 2D NMR methods. Dichotomins J (1) and K (2) showed a moderate vasorelaxant effect on rat aorta. PMID- 16309327 TI - N(3),5'-cycloxanthosine, the first natural occurrence of a cyclonucleoside. AB - An Eryus sp. of marine sponge from the Great Australian Bight has yielded the first reported natural occurrence of a cyclonucleoside, N(3),5'-cycloxanthosine. The structure of N(3),5'-cycloxanthosine was confirmed by detailed spectroscopic analysis and total synthesis. PMID- 16309328 TI - Cytotoxic prenylflavonoids from Artocarpus elasticus. AB - Five new prenylated flavonoids, artelastoheterol (1), artelasticinol (2), cycloartelastoxanthone (3), artelastoxanthone (4), and cycloartelastoxanthendiol (5), along with five known compounds, were isolated from the root bark of Artocarpus elasticus. The structures of 1-5 were elucidated by spectroscopic methods and through comparison with data reported in the literature. The previously known compound artonol A (6) exhibited cytotoxic activity against the A549 human cancer cell line, with an ED50 value of 1.1 microg/mL. PMID- 16309329 TI - Neoglycorandomization and chemoenzymatic glycorandomization: two complementary tools for natural product diversification. AB - In an effort to explore the contribution of the sugar constituents of pharmaceutically relevant glycosylated natural products, chemists have developed glycosylation methods that are amenable to the generation of libraries of analogues with a broad array of glycosidic attachments. Recently, two complementary glycorandomization strategies have been described, namely, neoglycorandomization, a chemical approach based on a one-step sugar ligation reaction that does not require any prior sugar protection or activation, and chemoenzymatic glycorandomization, a biocatalytic approach that relies on the substrate promiscuity of enzymes to activate and attach sugars to natural products. Since both methods require reducing sugars, this review first highlights recent advances in monosaccharide generation and then follows with an overview of recent progress in the development of neoglycorandomization and chemoenzymatic glycorandomization. PMID- 16309330 TI - The infrastructure of obesity and the obesity epidemic: implications for public policy. AB - If present increases in the rate of obesity persist, by 2015 half of the population in the US could be obese. This article presents the case that the reason for the epidemic is an 'infrastructure of obesity' that fuels the phenomenon. To control this epidemic, public policy in general and health policy in particular need to shift from market oriented policies that favour individual choice to policies that regulate the food supply, to help dismantle the infrastructure of obesity. Our research shows that state regulations have made some progress in the fight against obesity. PMID- 16309331 TI - The Chinese pharmaceutical market at the crossroads: pro-competition solutions to improve access, quality and affordability. AB - The Chinese pharmaceutical market is large in absolute size (the second largest in Asia), but it faces similar problems to those that plague other developing countries, such as a lack of adequate patent protection, low pharmaceutical spending per capita, but high pharmaceutical expenditure as a proportion of total medical spending, and a lack of health insurance coverage. In this article, the pros and cons of two pro-competition policy proposals for China are explored. The first proposal is to follow the E5 guideline of the International Conference on Harmonisation and waive unnecessary local clinical trials for global new drugs that have been approved by the US Food and Drug Administration or the European Medicines Agency (except for drugs for which there is a real possibility of there being ethnic differences in patient responses). The second proposal is to tighten the standards for generic drugs and approve only bioequivalent ones. While the first proposal encourages price competition between similar compounds in the same therapeutic class, the second proposal enhances generic competition for off patent drugs. Working together, these two proposals would improve access to and the quality and affordability of pharmaceuticals in China. PMID- 16309332 TI - Can privatising selected services benefit the public healthcare system?: the Greek case. AB - The evaluation of empirical data concerning private physician consultations in public hospitals in Greece suggests that privatisation of selected public services can increase patients' freedom of choice and also improve efficiency and quality of services, thus strengthening the supply side in a mixed healthcare system. The 2001 NHS health reform in Greece provides financial incentives to public hospital physicians to consult patients 'privately' at the outpatient clinics, while remaining restrained by certain provisions regarding fees. Competition between public and private hospitals is centrally regulated, and in this case cumulative data show success for the public sector. At the same time, patient satisfaction with the new arrangement measured in this study was very high. It could be argued that, under certain conditions, attracting private funds into a public health system constitutes an opportunity to reinforce the public provision of the system. PMID- 16309333 TI - Prisoner healthcare co-payment policy: a cost-cutting measure that might threaten inmates' health. AB - Cost-effective provision of adequate healthcare to prisoners is a core problematic issue in contemporary correctional healthcare settings. An increasingly popular policy for reducing prison healthcare costs is prisoner co payment systems for health services. Advocates of this policy assert that it facilitates efficient healthcare delivery in prison settings. This article examines the appropriateness and consequences of prisoner healthcare co-payment systems in US prisons. In conclusion, the policy has a strong potential to compromise prisoners' access to healthcare, while not significantly reducing prison healthcare costs. Alternative approaches for improving the efficiency of prison healthcare services are suggested, and implications for Australia are considered. PMID- 16309334 TI - Can we use contingent valuation to assess the demand for childhood immunisation in developing countries?: a systematic review of the literature. AB - Childhood immunisation is one of the most cost-effective public health interventions, yet its population coverage in low- and middle-income countries is severely limited by the fiscal constraints that health services face. A recent proposal suggested that commitments to purchase vaccines and make them available to developing countries for modest co-payments could solve the problem. However, this is dependent on communities being willing and able to share the cost in this way, which is difficult to assess. One possible method to assess this demand is contingent valuation (CV). This article evaluates the usefulness of using CV in this way, by reviewing applications of CV in developing countries against current 'standards' for CV of immunisation in the literature. A structured review was adopted with reference to the standard frameworks for methodological evaluation. A set of five criteria were developed for evaluating an 'acceptable' CV study: (i) response rate; (ii) association between willingness to pay (WTP) and socioeconomic status (SES); (iii) sensitivity of WTP to benefit scale/scope; (iv) predictive validity; and (v) reliability in elicitation formats. Two strands of literature search were conducted using electronic databases (MEDLINE, EMBASE, HEALTHSTAR and Econlit) from 1966 to 2003, one for CV studies of immunisation and one for CV studies in developing countries. Twelve CV studies of vaccination and 13 CV studies undertaken within developing countries were identified and reviewed. The quality of existing CV studies conducted in developing countries exceeded the benchmark standard set by studies of immunisation in the developed world in four of the five criteria. WTP estimates appeared both internally valid (i.e. associations with SES) and externally valid (i.e. predictive validity), reliability in developing countries was no less than that of the benchmark level in the existing literature, and the high response rates suggested that CV can be administered to a rural, and perhaps less literate, population. Only sensitivity to scale/scope was not well demonstrated. Our assessment indicated that the CV technique offers a promising tool to estimate the demand for childhood immunisation in low- and middle-income countries. International agencies are therefore encouraged to devote resources to such an application when designing their support to the immunisation programmes. PMID- 16309335 TI - Methodological challenges posed by economic evaluations of early childhood intervention programmes. AB - Early childhood intervention programmes have emerged in recent years with the aim of fostering the cognitive and social-emotional functioning and physical health of preschool children and enhancing their emerging competencies. This article presents a structured critical appraisal of economic evaluations of early childhood intervention programmes. It highlights a range of methodological issues in the field. These include: the fidelity of the evaluation process; the selection of the appropriate comparison group given the complexity of care routinely provided; the appropriate perspective and coverage of the study; methodological concerns relating to cost and benefit measurement and valuation; analytical requirements relating to the form of sensitivity analysis and the decision rules adopted by decision makers; and the interpretation of the results in the light of contextual factors. It is concluded that more transparent methodological guidance is required for analysts conducting economic evaluations of early childhood intervention programmes in particular and of public health interventions in general. Greater multidisciplinary collaboration between social scientists should also enhance the development of ground-breaking methods in this field. PMID- 16309336 TI - Barriers to the equitable funding of primary healthcare in South Africa. AB - Over 25 years after Alma-Ata, available evidence suggests that in low- and middle income countries (LMIC) the funding and coverage of primary care is still inequitable. This article reviews the progress that has been made towards the equitable funding of primary healthcare (PHC) in South Africa and evaluates barriers to future progress. The South African experience is assessed to consider implications for other LMIC. The results show that substantial inequities in funding persist (albeit using a narrow definition of PHC). Underlying causes relate to fiscal austerity, the lack of protection of PHC funding, incremental resource allocation and the belief that poorer districts are not able to use extra funds effectively. These results match the experience in other LMIC. Central governments have a critical role to play in lobbying for and protecting resources for PHC, maintaining more control over decentralised resource allocation and building capacity to benefit in poorer areas. PMID- 16309337 TI - Drug costs developments after patent expiry of enalapril, fluoxetine and ranitidine: a study conducted for the Netherlands. AB - BACKGROUND: In order to increase price competition, government regulations focus on controlling drug costs. Drug costs after patent expiry are an area of particular interest because the substitution of branded medication with generics represents an opportunity for lowering drug costs. However, drug costs may not decrease after patent expiry, because of a lack of price competition and different national pricing systems. AIM: The aim of this study was to investigate the trends in the use of generics after patent expiry for enalapril, fluoxetine and ranitidine and the subsequent changes, if any, in the costs of these medications. METHODS: A drug-utilisation study was performed using data from a large sample of Dutch pharmacies. Both volumes (measured as defined daily doses [DDD] per 1000 population) as well as drug costs (calculated per DDD) prior to and after patent expiry were calculated. Costs per DDD were compared using trend line analysis. In addition, the relative market shares of the different trade channels (branded, parallel imported and generic) were compared before and after patent expiry. RESULTS: The costs per DDD decreased for all three drugs and, as expected, these costs decrease more rapidly after patent expiry. Significant differences in the trend lines were found for enalapril and fluoxetine. CONCLUSIONS: Despite relatively high reimbursement prices for generics in the Netherlands, this example from the Dutch pharmaceutical market demonstrates the benefit of generic substitution for containing pharmaceutical costs, which contrasts with concerns raised by the Dutch government. PMID- 16309339 TI - [Not Available]. PMID- 16309340 TI - Los alamos hepatitis C immunology database. AB - The Los Alamos Hepatitis C Virus (HCV) Sequence Database (http://hcv.lanl.gov or http://hcv-db.org) was officially launched in September 2003. The sister HCV Immunology Database was made public in September 2004. The HCV Immunology Database is based on the Human Immunodeficiency Virus (HIV) Immunology Database. The HCV Immunology Database contains a curated inventory of immunological epitopes in HCV and their interaction with the immune system, with associated retrieval and analysis tools. This article describes in detail the types of data and services that the new database offers, the tools provided and the database framework. The data and some of the HCV database tools are available for download for non-commercial use. PMID- 16309341 TI - Feature Selection for Classification of SELDI-TOF-MS Proteomic Profiles. AB - BACKGROUND: Proteomic peptide profiling is an emerging technology harbouring great expectations to enable early detection, enhance diagnosis and more clearly define prognosis of many diseases. Although previous research work has illustrated the ability of proteomic data to discriminate between cases and controls, significantly less attention has been paid to the analysis of feature selection strategies that enable learning of such predictive models. Feature selection, in addition to classification, plays an important role in successful identification of proteomic biomarker panels. METHODS: We present a new, efficient, multivariate feature selection strategy that extracts useful feature panels directly from the high-throughput spectra. The strategy takes advantage of the characteristics of surface-enhanced laser desorption/ionisation time-of flight mass spectrometry (SELDI-TOF-MS) profiles and enhances widely used univariate feature selection strategies with a heuristic based on multivariate de correlation filtering. We analyse and compare two versions of the method: one in which all feature pairs must adhere to a maximum allowed correlation (MAC) threshold, and another in which the feature panel is built greedily by deciding among best univariate features at different MAC levels. RESULTS: The analysis and comparison of feature selection strategies was carried out experimentally on the pancreatic cancer dataset with 57 cancers and 59 controls from the University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania, USA. The analysis was conducted in both the whole-profile and peak-only modes. The results clearly show the benefit of the new strategy over univariate feature selection methods in terms of improved classification performance. CONCLUSION: Understanding the characteristics of the spectra allows us to better assess the relative importance of potential features in the diagnosis of cancer. Incorporation of these characteristics into feature selection strategies often leads to a more efficient data analysis as well as improved classification performance. PMID- 16309342 TI - A robust statistical method for detecting differentially expressed genes. AB - DNA microarray technology allows researchers to monitor the expressions of thousands of genes under different conditions, and to measure the levels of thousands of different DNA molecules at a given point in the life of an organism, tissue or cell. A wide variety of different diseases that are characterised by unregulated gene expression, DNA replication, cell division and cell death, can be detected early using microarrays. One of the major objectives of microarray experiments is to identify differentially expressed genes under various conditions. The detection of differential gene expression under two different conditions is very important in biological studies, and allows us to identify experimental variables that affect different biological processes. Most of the tests available in the literature are based on the assumption of normal distribution. However, the assumption of normality may not be true in real-life data, particularly with respect to microarray data.A test is proposed for the identification of differentially expressed genes in replicated microarray experiments conducted under two different conditions. The proposed test does not assume the distribution of the parent population; thus, the proposed test is strictly nonparametric in nature. We calculate the p-value and the asymptotic power function of the proposed test statistic. The proposed test statistic is compared with some of its competitors under normal, gamma and exponential population setup using the Monte Carlo simulation technique. The application of the proposed test statistic is presented using microarray data. The proposed test is robust and highly efficient when populations are non-normal. PMID- 16309343 TI - Design of large time-course microarray experiments with two channels. AB - In this article we propose two practical types of designs for large time-course, dual-channel microarray experiments. One type consists of several interwoven loops, and the other type combines reference and loop designs. By representing the experiment as a graph, where the timepoints are nodes and the arrays are edges, we demonstrate how the time contrasts between any two timepoints can be estimated, provided that there is a path of edges linking them. In addition, we give a general formula for the variance of such contrasts. The efficiency of the proposed designs is evaluated by estimating the variances of the log-ratios of the comparisons of interest. PMID- 16309344 TI - Random walk models for bayesian clustering of gene expression profiles. AB - The analysis of gene expression temporal profiles is a topic of increasing interest in functional genomics. Model-based clustering methods are particularly interesting because they are able to capture the dynamic nature of these data and to identify the optimal number of clusters. We have defined a new Bayesian method that allows us to cope with some important issues that remain unsolved in the currently available approaches: the presence of time dislocations in gene expression, the non-stationarity of the processes generating the data, and the presence of data collected on an irregular temporal grid. Our method, which is based on random walk models, requires only mild a priori assumptions about the nature of the processes generating the data and explicitly models inter-gene variability within each cluster. It has first been validated on simulated datasets and then employed for the analysis of a dataset relative to serum stimulated fibroblasts. In all cases, the results have been promising, showing that the method can be helpful in functional genomics research. PMID- 16309345 TI - BioCloneDB: A Database Application to Manage DNA Sequence and Gene Expression Data. AB - BioCloneDB is a user-friendly database with a web interface to assist molecular genetics laboratories in managing a local repository of sequence information linked to DNA clones. This tool is designed to assist in high-throughput sequence and gene expression projects, providing a link between both types of information. The unique feature of the application is the automation of batch sequence annotation following BLAST((R)) searches, which is supported by easy-to-use web interfaces. Furthermore, any set of sequences can be annotated against any sequence database. This replaces the need to perform and analyse individual web BLAST((R)) searches or the need to learn how to produce batch searches and perform analysis in a UNIX((R)) operating system. BioCloneDB is open-source software that can be installed on Linux or UNIX((R)) operating systems. To test the application, we used 1400 expressed sequence tags obtained from the filamentous fungus Neurospora crassa. The results were analysed and compared with published results and they show a significant change due to the accumulation of the data in the nr database (ftp://ftp.ncbi.nih.gov/blast/db/). AVAILABILITY: BioCloneDB is available for academic use along with documentation, screenshots, database scheme and readme files at http://bioclonedb.agri.huji.ac.il/ CONTACT: Oded Yarden (Oded.Yarden@huji.ac.il). PMID- 16309346 TI - GOAT: An R Tool for Analysing Gene Ontologytrade mark Term Enrichment. AB - Understanding the composition of gene lists that result from high-throughput experiments requires elaborate processing of gene annotation lists. In this article we present GOAT (Gene Ontology Analysis Tool), a tool based on the statistical software 'R' for analysing Gene Ontologytrade mark (GO) term enrichment in gene lists. Given a gene list, GOAT calculates the enrichment and statistical significance of every GO term and generates graphical presentations of significantly enriched terms. GOAT works for any organism with a genome-scale GO annotation and allows easy updates of ontologies and annotations. AVAILABILITY: GOAT is freely available from http://dictygenome.org/software/GOAT/ CONTACT: Gad Shaulsky (gadi@bcm.tmc.edu). PMID- 16309347 TI - [In vitro activity of the echinocandins. How should it be evaluated?]. AB - Echinocandins are a novel class of antifungal drugs. They have good activity against Candida spp and Aspergillus spp. Their low selective toxicity allows their administration at high doses with few secondary side effects. We have reviewed the available data on the endpoints for these drugs in their in vitro susceptibility testing on yeasts and moulds. The microdilution broth method is the most commonly used technique and MIC-1 (80% of growth inhibition) seems to be the most reliable endpoint when yeasts are tested. This endpoint also seems to be the most appropriate for the different drugs when they are combined with echinocandins using the checkerboard method for testing yeasts. By contrast, in the case of moulds, the minimum effective concentration (MEC) correlates better with the in vivo activity than the MIC when echinocandins are tested, and when these drugs are combined with other antifungals, MIC-2 (50% of growth inhibition) seems the most appropriate endpoint. Criteria based on drug pharmacodynamics is the most useful to define the echinocandin endpoints that best correlate with their in vivo efficacy. PMID- 16309348 TI - Histoplasmosis in a Brazilian center: clinical forms and laboratory tests. AB - Histoplasmosis, caused by the dimorphic fungus Histoplasma capsulatum, is endemic in many regions of the Americas, Asia and Africa. It has a wide spectrum of clinical manifestations, from asymptomatic infection to severe disseminated disease. A retrospective study was carried out to describe the clinical forms and assess the clinical significance of the laboratory diagnostic tests of patients with histoplasmosis during the period of July 1987 to December 2003 at Instituto de Pesquisa Clinica Evandro Chagas/ FIOCRUZ, RJ, Brazil. Seventy-four patients were included. Forty-nine percent of the cases (n = 36) occurred in HIV positive patients who presented with disseminated disease. The remaining 38 cases were classified in different clinical forms. Histoplasma capsulatum was isolated from 69.5% of the clinical specimens sent to culture. Immunodiffusion and immunoblot were positive in 72.6% and 100% of the performed tests, respectively. Histopathologic findings suggestive of H. capsulatum were found in 63.2% of the performed exams. Serology had a lower proportion of positivity amongst AIDS patients, when compared with HIV negative patients (X2 = 6.65; p lower than 0.008). Statistical differences between AIDS and non-AIDS patients were not observed with culture and histopathology. The specific role of each test varies according to the clinical form. Physicians need to know the value and limitations of the available diagnostic tests, but before that, they have to think about histoplasmosis and consider this clinical entity in their differential diagnosis. PMID- 16309349 TI - [Importance of the sporotrichosis asteroid body for the rapid diagnosis of sporotrichosis]. AB - Eighty patients with cutaneous sporotrichosis were studied between 1985 and 1996. The investigation of asteroid bodies (AB) was done by direct microscopic slides examination of pus obtained from the lesions. Patients were divided into two groups: 32 consulting before 1989, and 48 consulting after 1990. In the first group, material was obtained as usual by simple digital pressure of the most productive lesion, and then wet preparation microscopic examination was performed. Fourteen patients with AB were found in this group (43.75%). In the second group the initial pus was discarded and new samples were taken more deeply, and examined up to five slides for each patient. The slides were carefully examined at light microscope. Fourty five patients with AB were detected in this group (93.75%). All eighty samples were cultured and all of them were positive for Sporothrix schenckii. The change of methodology to obtain the samples and the exhaustive observations, increased the possibility of AB detection. ABs are of great diagnostic value and might be of importance to initiate treatment before reporting culture. PMID- 16309350 TI - [Physiological comportment and in vivo sensitivity of Sporothrix schenckii isolates maintained for 18 years by two preservation methods]. AB - We compared two methods for the preservation of fungi, Castellani's method and repeated passage in Sabouraud medium-agar, on five isolates of Sporothrix schenckii that were preserved for 18 years at room temperature by both procedures. They were evaluated for viability of the strains, growth rate, morphological and physiological characteristics, and in vitro sensitivity to iodide, itraconazole, terbinafine and posaconazole. 100% viability was observed in all of the isolates, with slower growth rate on strains preserved in water compared to strains periodically re-cultured. The typical morphological feature of these fungi was preserved by both methodologies. With regard to enzymatic activity, both groups gave urease reactions and were beta glucosidase-positive. Nevertheless, complete inhibition of the capacity to hydrolyse starch was observed only on the isolates preserved in water. This group also was more sensitive to potassium iodide at a concentration of 10 microM in the in vitro sensitivity tests. PMID- 16309351 TI - Could Candida dubliniensis be involved in lung fungus balls? AB - We describe a case of cavitary pneumonia due to Candida dubliniensis along with fungemia due to Candida kefyr in a leukemic patient. This is the first case of C. dubliniensis isolated in our laboratory. The identification was performed by phenotypic and molecular methods such as thermotolerance test, carbohydrate fermentation and polymerase chain reaction. PMID- 16309352 TI - [Aspergillus fumigatus in nasopharyngeal cavity of horses]. AB - Aspergillus fumigatus is a cosmopolitan opportunistic fungal associated to rhinopharyngitis, sinusitis and guttural pouches infection with nasal discharges. All them are similar with Strangle's sign, the infectious disease produced by Streptococcus equi spp. The aim of this work was to detect A. fumigatus in healthy horses living in boxes and field. 226 nasopharyngeal swabbing samples were obtained by mycological routine. A. fumigatus was isolated in 26 (11.5%) horses. PMID- 16309353 TI - [Revisiting the epidemiology of onychomycoses]. AB - Onychomycoses represent a group of nail affections caused by one or more fungi. Whether aging represents a risk factor for developing onychomycosis, remains a question. In the present work, we studied the variations in frequency and prevalence of onychomycoses and non-mycotic onychodystrophies according to age. Our results show that there is an increase in the frequency and prevalence of onychomycoses and non-mycotic onychodystrophies predominantly in patients over forty years of age. PMID- 16309354 TI - Sparing of the upper axillary area in pityriasis versicolor. AB - Increased temperature and sweating are considered factors predisposing to pityriasis versicolor. In this case report, sparing of the vaults of the axillae which are naturally occluded and sweaty areas, in a patient with widespread lesions of pityriasis versicolor is discussed. It is the second observation in this regard and further contributions are invited. PMID- 16309355 TI - Imported and autochthonous histoplasmosis in Italy: new cases and old problems. AB - In the past the Italian soil was considered as a low-endemic pabulum for H. capsulatum var. capsulatum and only few autochthonous cases of histoplasmosis were reported in Italy, especially in the Po valley. The aim of the paper was to evaluate this possibility by reviewing the literature and providing our own personal data. Four additional cases of histoplasmosis were observed during 1999 2003 in AIDS immigrant or in Italian citizens, and in travellers to endemic areas. One of the AIDS patients was an autochthonous case of histoplasmosis. The Italian literature was reviewed. Recent cases and literature data confirm the possible autochthonous presence of histoplasmosis in Italy, especially in the Northern regions. PMID- 16309356 TI - [Scytalidium dimidiatum an opportunistic fungus for both man and Mangifera indica trees in Venezuela]. AB - The purpose of this study was to confirm the presence of Scytalidium dimidiatum on Mangifera indica (mango) trees, in a plantation managed by a diabetic patient with a white grain mycetoma of the foot caused by the same fungus. Samples from necrotic apices, roots, burned leaves and rotten stems from eight trees were processed by the Smith and Furcolow's mineral oil technique (modified). Several isolates from the apex material and clinical samples from the diabetic patient isolated in pure culture a fungus with the macroscopic and microscopic characteristics of that in S. dimidiatum. This fungus should be considered as an opportunistic microorganism for both humans and M. indica. PMID- 16309357 TI - Sexual reproduction in subcultures of Absidia blakesleeana after years of preservation under mineral oil. AB - Two subcultures of Absidia blakesleeana IOC No. 2425 (-) mating type strain preserved in 1959 and 1981 on a thick layer of potato dextrose agar (PDA) medium under sterile mineral oil and maintained at room temperature in the Fungal Cultural Collection of Institute Oswaldo Cruz--IOC--were paired with the defined Absidia blakesleeana URM-UFP No. 2076 (+) mating type strain. Portions of the line where mycelia of the two strains met were observed under light microscopy. Zygospores belonging to Subgenus Mycocladus were observed. The results demonstrated conservation and re-establishment of the physiological and genetic metabolic processes of two subcultures preserved under mineral oil. PMID- 16309358 TI - [Clinical cases in medical mycology. Case No. 18]. PMID- 16309359 TI - [Clinical cases in medical mycology. Case No. 19]. PMID- 16309360 TI - Tinea faciei in a goat handler due to Microsporum canis. PMID- 16309361 TI - Role of Geotrichum candidum in canine oral ulcers. PMID- 16309362 TI - ProteinChips: the essential tools for proteomic biomarker discovery and future clinical diagnostics. PMID- 16309363 TI - Protein profiling as a diagnostic tool in clinical chemistry: a review. AB - Serum protein profiling by surface-enhanced laser desorption/ionization time-of flight mass spectrometry (SELDI-TOF MS) appears to be an important diagnostic tool for a whole range of diseases. Sensitivities and specificities obtained with this new technology often seem superior to those obtained with current biomarkers. However, reproducibility and standardization are still problematic. The present review gives an overview of the diagnostic value of protein profiles obtained with SELDI in studies on prostate and ovarian cancer. To identify aspects important for protein profiling, we compare and discuss differences in pre- and post-analytical conditions presented in the literature supplemented with some of our own data. Further progress in protein profiling as a diagnostic tool requires a more comprehensive description of technical details in all future studies. PMID- 16309364 TI - Protein biochip systems for the clinical laboratory. AB - Classical methods of protein analysis such as electrophoresis, ELISA and liquid chromatography are generally time-consuming, labor-intensive and lack high throughput capacity. In addition, all existing methods used to measure proteins necessitate multiple division of the original sample and individual tests carried out for each substance, with an associated cost for each test. The chip system allows several tests to be performed simultaneously without dividing the original patient sample. This system facilitates the development of multiplexed assays that simultaneously measure many different analytes in a small sample volume. These emerging technologies fall into two categories: 1) spotted array-based tools, and 2) microfluidic-based tools. Miniaturized and multiplexed immunoassays allow a great deal of information to be obtained from a single sample. These analytical systems are referred to as "lab-on-a-chip" devices. This article presents current trends and advances in miniaturized multiplexed immunoassay technologies, reviewing different systems from research to point-of-care assays. We focus on a subset of chip-based assays that may be used in a clinical laboratory and are directly applicable for biomedical diagnosis. Recent advances in biochip assays combine the power of miniaturization, microfluidics, micro- to nanoparticles, and quantification. A number of applications are just beginning to be explored. The power of biochip assays offers great promise for point-of-care clinical testing and monitoring of many important analytes. PMID- 16309365 TI - Automation of biochip array technology for quality results. AB - BACKGROUND: Proteomics' requirement for simultaneous measurement of multiple markers is now possible with biochip array technology. Many laboratories utilise in-house, manual procedures for biochip fabrication and sample testing. Reproducibility and standardisation of biochip processes is vital to ensure quality of results and offer the best tool for elucidation of complex relationships between multiple proteins in diseased conditions. METHODS: Various novel control checks have been implemented in biochip fabrication, reagent manufacture, automation and imaging processes for the Evidence analyser. Reference spots enable location of discrete test regions on the surface of the biochip and simultaneous quantification of multiple markers. Performance and standardisation methods are presented. RESULTS: Formulation of dispense solution for discrete test regions had a direct effect on their shape, stability and integrity on the biochip surface. Assays for fertility hormones and drugs of abuse demonstrated excellent precision, stability and comparison with other commercial methods. CONCLUSION: Control processes employed in the manufacture and analysis of Evidence components ensure reproducibility of assays for a range of routine and novel markers. PMID- 16309366 TI - SELDI-TOF-MS proteomics of breast cancer. AB - The detection, diagnosis, and management of breast cancer rely on an integrated approach using clinical history, physical examination, imaging, and histopathology. The discovery and validation of novel biomarkers will aid the physician in more effectively achieving this integration. This review discusses efforts in surface-enhanced laser desorption/ionization (SELDI)-based proteomics to address various clinical questions surrounding breast cancer, including diagnosis, monitoring, and stratification for treatment. Emphasis is placed on examining how study design and execution influence the discovery and validation process, which is critical to the proper development of potential clinical tests. PMID- 16309367 TI - Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development. AB - In the emerging field of Functional Proteomics, protein microarrays are considered to be one of the most promising tools for the simultaneous analysis of the a) abundance, b) function, and c) interaction of proteins on a system-wide scale. Resting on the technological grounds of widely used DNA biochips, the great power of microarray-based miniature solid-phase immunoassays lies in their potential to investigate in parallel large numbers of analyte pairs in a variety of biological samples. Consequently, this has fueled aspirations that protein microarrays may serve as tools for the high-throughput functional investigation of complete proteomes and, moreover, that they will develop into promising candidates for innovative in-vitro diagnostic (IVD) applications. To date, published examples of protein microarrays for IVD purposes have included tests for allergy, autoimmune and infectious diseases. Here, we discuss recent advancements in the development of protein microarrays for the profiling of IgE antibodies in the diagnosis of Type 1-related allergic diseases. PMID- 16309368 TI - Separation of human serum proteins using the Beckman-Coulter PF2D system: analysis of ion exchange-based first dimension chromatography. AB - The ProteomeLab PF2D protein fractionation system is a rapid, semi-automated, 2 D HPLC instrument that uses two different methods to separate plasma serum proteins: ion-exchange chromatography using a wide pH range in the first dimension, and non-porous reverse-phase chromatography in the second dimension. Because this methodology has only very recently been introduced in proteomic laboratories, little is known about the characteristics of PF2D fractionation of human serum proteins. To evaluate the system's application in a clinical laboratory setting, the characteristics of the ion-exchange chromatography-based separation were analyzed. Following fractionation of human serum proteins on a linear pH gradient (ranging from 8.0 to 4.0), each fraction was collected in a cool module of the instrument. Different fractions obtained from the first dimension were then pooled together and loaded on classic 2D gel electrophoresis instrumentation. The different spots obtained were then checked against the Swiss Prot Data-base. A total of 36 human serum proteins were identified in different PF2D-generated fractions. Some important features of the separation system were observed. Different eluted fractions contained different proteins, thus demonstrating the reliability of the fractionation system. The proteins were also fractionated according to the theoretical isoelectric point (pI). This was consistent with the evidence that the vast majority of immunoglobulins, characterized by an alkaline pI, were not retained by the column and were eluted in the unbound fraction. This outcome also underlines a practical advantage: fractions eluted from pH 8 to pH 4 contained virtually immunoglobulin-depleted serum proteins. This finding supports an immediate use of the PF2D system in a clinical setting, where abundant proteins should be clearly identified to enable evaluation of other less abundant, but potentially relevant, species. PMID- 16309369 TI - Rapid, accurate genotyping of alcohol dehydrogenase-1B and aldehyde dehydrogenase 2 based on the use of denaturing HPLC. AB - The genotypes of alcohol dehydrogenase-1B (ADH1B) and aldehyde dehydrogenase-2 (ALDH2) are related to alcohol dependence and some human disorders. Rapid, accurate genotyping methodologies for specific polymorphisms of these two genes are needed for molecular screening and testing of alcohol-related problems in populations. A polymerase chain reaction (PCR) composed of two separate amplicons was designed to generate sequences containing the polymorphic site of interest in the ADH1B and ALDH2 genes. The PCR amplicons for each sample were subjected to denaturing high-performance liquid chromatography (DHPLC), analysis performed under partially denaturing conditions as determined by profiling the mixture of a unique homozygous control and a tested sample amplicon. A total of 150 genomic DNA samples were tested to validate this assay by blind analysis. Direct DNA sequencing was performed on samples randomly selected from each of the genotype groups detected by DHPLC profiling. The results indicate 100% concordance between the sequencing analysis and the DHPLC detection. The method we present provides a reliable and fast genotyping procedure for molecular screening of alcohol-related problems. PMID- 16309370 TI - APOA1 polymorphisms are associated with variations in serum triglyceride concentrations in hypercholesterolemic individuals. AB - BACKGROUND: Apolipoprotein A-I gene (APOA1) polymorphisms have been associated with variations in serum low-density lipoprotein (LDL)- and high-density lipoprotein (HDL)-cholesterol. We have investigated whether APOA1 common variants are also associated with variations in basal triglyceride serum concentrations and response to atorvastatin in individuals with hypercholesterolemia. METHODS: APOA1 G-75A and C83T polymorphisms and variations in serum lipids were evaluated in 150 hypercholesterolemic (HC) and 93 normolipidemic (NL) unrelated European derived Brazilians treated with atorvastatin (10 mg/day for 4 weeks). Genomic DNA was extracted from blood leukocytes using a salting-out method and APOA1 polymorphisms were analyzed by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: G-75A polymorphism was associated with differences in serum concentrations of triglyceride and very low-density lipoprotein (VLDL) cholesterol (p=0.026) in HC men. After atorvastatin treatment, women carrying the GG/CC haplotype had lower serum triglyceride and VLDL-cholesterol (p=0.020) than non-carriers. In men, the reduction in serum triglyceride in response to atorvastatin was found to be slightly lower in GG/CC haplotype carriers (p=0.051). CONCLUSION: Our data suggest that APOA1 polymorphisms are associated with variations of baseline serum concentrations of triglyceride and VLDL cholesterol and in response to atorvastatin in a gender-specific manner. PMID- 16309371 TI - Simple PCR heteroduplex, SSCP mutation screening methods for the detection of novel catalase mutations in Hungarian patients with type 2 diabetes mellitus. AB - BACKGROUND: The enzyme catalase is the main regulator of hydrogen peroxide metabolism. Deficiency of catalase may cause high concentrations of hydrogen peroxide and increase the risk of the development of pathologies for which oxidative stress is a contributing factor, for example, type 2 diabetes mellitus. Catalase deficiency has been reported to be associated with increased frequency of diabetes mellitus in a cohort of patients in Hungary. In this cohort, the majority of mutations in the catalase gene occur in exon 2. METHODS: Type 2 diabetic patients (n=308) were evaluated for mutations in intron 1 (81 bp), exon 2 (172 bp) and intron 2 (13 bp) of the catalase gene. Screening for mutations utilized PCR single-strand conformational polymorphism (SSCP) and PCR heteroduplex methods. Verification of detected mutations was by nucleotide sequence analysis. RESULTS: A total of 11 catalase gene mutations were detected in the 308 subjects (3.57%, p<0.001). Five of the 11 were at two previously reported mutation sites: exon 2 (79) G insertion and (138) GA insertion. Six of the 11 were at five previously unreported catalase mutation sites: intron 1 (60) G-->T; intron 2 (7) G-->A and (5) G-->C; exon 2 (96) T-->A; and exon 2 (135) T- >A. The novel missense mutations on exon 2 (96 and 135) are associated with 59% and 48% decreased catalase activity, respectively; the novel G-->C mutation on intron 2 (5) is associated with a 62% decrease in catalase activity. Mutations detected on intron 1 (60) and intron 2 (7) showed no change in catalase activity. The G-->C mutation on intron 2 (5) might be a splicing mutation. The two missense mutations on exon 2 (96) and (135) cause substitutions of amino acids 53 (Asp- >Glu) and 66 (Glu-->Cys) of the catalase protein. These are close to amino acids that are important for the binding of heme to catalase, 44 (Val) and 72-75 (Arg, Val, Val, His). Changes in heme binding may be responsible for the activity losses. CONCLUSION: Mutations that cause decreased catalase activity may contribute to susceptibility to inherited type 2 diabetes mellitus. Exon 2 and neighboring introns of the catalase gene may be minor hot spots for type 2 diabetes mellitus susceptibility mutations. PMID- 16309372 TI - Glycogen phosphorylase BB in acute coronary syndromes. AB - The diagnosis of myocardial damage is preferably based on measurement of the cardiac-specific troponins. However, there is an emerging need for early, specific cardiac markers. One potential candidate is the glycogen phosphorylase BB isoenzyme (GPBB). We investigated the use of a new, commercially available GPBB ELISA assay in 61 patients presenting with an acute coronary syndrome (37 acute myocardial infarction, 24 unstable angina pectoris) in comparison to established cardiac markers such as troponin T, creatine kinase isoenzyme MB (CKMB) mass, and myoglobin. Blood samples were obtained on arrival, as well as 1, 2, 3, 4, 8, 12 and 24 h later. GPBB plasma concentrations were elevated in 90.9% of patients 1 h after onset of chest pain and increased to 100% at 4-5 h. Within the first 6 h, GPBB showed the highest sensitivity (95.5-100%) and high specificity (94-96%) compared to myoglobin (85-95% sensitivity) and CKMB mass (71.4-91.3% sensitivity). As expected, troponin T showed high specificity (100%) and sensitivity >95% later in the time course (>or=3 h). In un-stable angina pectoris patients, a very high rate of elevated GPBB was observed (93.9% at 3 h) compared to myoglobin (66.7%). Cardiac troponin T and CKMB were only elevated in 33.8% and 55.0% of these patients, respectively. In conclusion, GPBB is a promising marker for the early diagnosis of acute coronary syndromes and could probably act as a marker of ischemia. However, further studies on specificity and development of a fast, automated assay are necessary before GPBB can be recommended as a routine diagnostic tool. PMID- 16309373 TI - Alteration in serum leptin correlates with alterations in serum N-telopeptide of collagen type I and serum osteocalcin during the progression of osteoporosis in ovariectomized rats. AB - The role of leptin during the progression of osteoporosis was investigated in ovariectomized rats by correlation of serum leptin levels with N-telopeptide of collagen type I (NTx) and osteocalcin levels before ovariectomy and 20, 40 and 60 days after the operation. Furthermore, peripheral quantitative computed tomography was used to confirm the development of severe osteoporosis in rats on day 60. The levels of NTx and osteocalcin were significantly increased on day 20 [61.9+/-5.4 nM BCE (bone collagen equivalents) and 215.6+/-53.3 ng/mL, respectively] in comparison to those before ovariectomy (41.3+/-1.7 nM BCE and 60.4+/-10.9 ng/mL). Accordingly, leptin was significantly elevated on day 20 (3033+/-661 vs. 606+/-346 pg/mL before ovariectomy). Bone markers and leptin levels remained constant up to day 40, while a slight, but not statistically significant, decrease was noted for osteocalcin and leptin on day 60. Although leptin and bone markers did not correlate before ovariectomy (r=0.09 for NTx and r=-0.05 for osteocalcin), strong correlation was observed at all time points after ovariectomy. The data obtained suggest that the alterations in serum leptin levels during the progression of osteoporosis in ovariectomized rats follow the alterations in bone markers. PMID- 16309374 TI - Glycemic control in diabetes in three Danish counties. AB - BACKGROUND: Hemoglobin A1c (HbA1c) is a proxy measure for glycemic control in diabetes. We investigated the trend for glycemic control in patients from three Danish counties using HbA1c measurements. METHODS: We studied 2454 patients from a population of 807,000 inhabitants for whom routine monitoring of diabetes using HbA1c-DCCT aligned was initiated in 2001. We estimated the incidence of monitored patients in the population. The progress in patients with originally diabetic HbA1c levels was investigated by cumulative probability plots, and the individual trend in clinical outcome was investigated by a modified difference plot. RESULTS: The age-standardized incidence of monitored patients was <0.5% in all regions. Patients with diabetic first HbA1c concentrations (>or=6.62% HbA1c) showed on average 15% improved glycemic control in the first year. Further improvement was limited. The overall percentage above the treatment target (>or=6.62% HbA1c) was 51% in 2003 compared to 59% in 2001, and the percentage with poor glycemic control (>or=10.0% HbA1c) was reduced from 19% to 4%. Of patients with originally diabetic HbA1c levels, 15% showed progress in glycemic control, and 28% reached treatment targets. In patients with originally normal HbA1c, 75% showed an upward trend in HbA1c levels, which reached diabetic concentrations in 17%. CONCLUSION: Patients with diabetic first HbA1c concentrations (>or=6.62% HbA1c) showed on average 15% improved glycemic control in the first year. Further improvement was limited. In individual patients, 75% with originally diabetic HbA1c levels showed improved glycemic control after 3 years, while 78% with originally normal concentrations showed an upward trend in HbA1c levels. PMID- 16309375 TI - Atorvastatin suppresses homocysteine formation in stimulated human peripheral blood mononuclear cells. AB - Hyperhomocysteinemia is regarded as an independent risk factor for vascular diseases, and homocysteine is supposed to contribute to oxidative stress and endothelial damage. Statin therapy is an established intervention to reduce the risk of acute events in patients suffering from cardiovascular diseases. Apart from their lipid-lowering capacity, statins also exert anti-inflammatory and antioxidant effects. As cellular immune activation and oxidative stress play a major role in the pathogenesis of cardiovascular diseases, the anti-inflammatory capacity of statins could partly be responsible for the beneficial effects observed in patients. Earlier we reported that stimulated peripheral blood mononuclear cells (PBMCs) release homocysteine. Here we studied the influence of atorvastatin on homocysteine production in stimulated PBMCs and compared changes in cysteine concentrations and in neopterin production, which is a sensitive indicator of cellular immune activation. Stimulation of human PBMCs with the mitogens concanavalin A and phytohemagglutinin induced significant homocysteine and neopterin production compared to unstimulated cells, whereas cysteine concentrations remained unchanged. Treatment of PBMCs with increasing doses of atorvastatin (10-100 microM) suppressed both biochemical pathways in a dose dependent manner, and cell proliferation was inhibited in parallel. Again, cysteine levels were not influenced by any treatment. The down-regulating effect of atorvastatin on homocysteine formation in vitro indicates that statins may prevent homocysteine accumulation in the blood via immunosuppression. PMID- 16309376 TI - Buprenorphine detection in biological samples. AB - Buprenorphine is a narcotic analgesic used in the treatment of moderate-to-severe pain. In Italy, buprenorphine is now being used as an alternative in the therapeutic treatment of heroin abusers. Even if confirmation methods are needed for identification and quantitation, a rapid and sensitive test, such as an immunoassay, is essential for screening. We evaluated the Buprenorphine One-step ELISA test (designed for serum specimens) for the qualitative determination of buprenorphine in other matrices, such as urine, saliva and hair. PMID- 16309377 TI - The effect of thyroid antibody positivity on reference intervals for thyroid stimulating hormone (TSH) and free thyroxine (FT4) in an aged population. AB - Our aims were: 1) to analyze the effect of the methodology used to derive clinically feasible cut-off values for thyroid peroxidase antibodies (TPOAb) and thyroglobulin antibodies (TgAb), which exhibit highly skewed distributions; and 2) to describe the influence of thyroid antibodies on thyroid stimulating hormone (TSH) and free thyroxine (FT4) reference intervals among thyroid disease-free aged subjects. The reference population consisted of 1086 individuals with a mean age of 73 years. The impacts of TPOAb and/or TgAb positivity on the reference intervals of TSH and FT4 were evaluated by both including and excluding subjects with elevated thyroid antibodies. The exclusion of subjects with elevated thyroid antibodies had no effect on the FT4 reference interval in either gender or on the TSH reference interval in men. Among women, the exclusion of 196 (34%) thyroid antibody-positive subjects resulted in lowering of the upper reference limit of TSH from 7.2 to 5.8 mIU/L. When the more stringent "mode-method" by summing mode+(mode-2.5th percentile) was used, 334 women (58%) were excluded and the upper reference limit of TSH remained essentially identical. Regardless of the statistical methodology used to derive cut-off values, the effect of antibody positivity was found to be less than expected. PMID- 16309378 TI - High-affinity antibodies in a new immunoassay for plasma tissue factor: reduction in apparent intra-individual variation. AB - Tissue factor, the main initiator of blood coagulation, is shed into plasma by blood cells and endothelium. While studying such circulating plasma tissue factor with a commercially available immunoassay, we found unsatisfactory results and therefore developed a new and highly sensitive enzyme-linked immunosorbent assay (ELISA). High-affinity monoclonal antibodies raised against recombinant soluble tissue factor were used and the new assay had a detection limit of 40 fmol/L, approximately six-fold lower than existing assays. Normal ranges in 20 healthy donors were established in serum and in citrated EDTA and heparinized plasma. Tissue factor was also measured in three successive plasma samples from 43 patients with type 2 diabetes mellitus. In citrated plasma from healthy donors, tissue factor concentrations were 2.5 (1.0-9.3) pmol/L (median with range) and were not significantly different in diabetics. With a commercially available immunoassay, seven plasma samples were below the detection limit. Use of the new assay reduced intra-individual variation in diabetics from 49% to 14% and we conclude that high-affinity antibodies may markedly improve immunoassay performance. PMID- 16309380 TI - Sensitive immunoassays for the autoantibodies reacting against citrullinated carboxy-terminal telopeptides of type I and type II collagens in patients with rheumatoid arthritis. AB - We developed sensitive assay methods for autoantibodies recognizing the citrullinated synthetic peptides derived from type I and type II collagens in patients with rheumatoid arthritis (RA). These peptides were tested with the chemiluminescence method (Nichols Advantage System). In 44 RA patients out of 120, the sera showed increased binding of citrullinated synthetic C-telopeptide derived from the alpha1 chain of type I collagen (p=0.003 compared to controls). For a corresponding C-telopeptide pair from the alpha1 chain of type II collagen, 35 patient sera bound the citrullinated peptide more strongly than the arginine peptide, but the difference compared to the controls was not significant (p=0.074). Correlation between the two carboxy-telopeptides was r=0.473 (p<0.001). The anti-CCP assay (antibodies against citrullinated filaggrin sequence-derived peptides) was positive in 59% of our RA patients. There was no relationship between the anti-CCP results and the antibodies against collagen C telopeptides, but both are increased in RA patients. We demonstrated autoantibodies in RA patients that bound citrullinated C-telopeptides derived from type I and type II collagen antigens. The peptide sequences detected (-YYXA and -YMXA) were different from that based on the cyclic filaggrin antigen (-STXG , where X represents citrulline). PMID- 16309379 TI - Physiological matrix metalloproteinase concentrations in serum during childhood and adolescence, using Luminex Multiplex technology. AB - Matrix metalloproteinases are a family of zinc-dependent proteinases which are involved in the breakdown and remodeling of extracellular matrix. As children grow and adolescents reach pubescence, their bodies undergo changes that require age-related morphogenesis of the extracellular matrix, possibly requiring unique patterns of matrix metalloproteinase (MMP) expression during periods of rapid tissue growth (i.e., childhood) or accelerated tissue remodeling and expansion (i.e., adolescence). Therefore, we have characterized age-specific and gender specific differences in circulating concentrations of MMPs (specifically MMP-1, 2, -3, -8 and -9) in 189 serum samples obtained from healthy subjects, aged 2-18 years. MMP concentrations were measured using Fluorokine MultiAnalyte Profiling kits and a Luminex Bioanalyzer, as well as by commercial ELISA. Serum levels of MMP-1, -2, -3, -8, and -9 in healthy pediatric subjects represent log-normal distributions. MMP-2 was significantly negatively correlated with age (r=-0.29; p<0.001), while MMP-3 was significantly positively correlated with age (r=0.38; p<0.001). Although plasma, not serum, is considered the appropriate blood sample for measurement of MMP-8 and -9, serum levels of MMP-8 and -9 were also found to be highly positively correlated with each other (r=0.76; p<0.01). MMP results obtained by Fluorokin MultiAnalyte Profiling methods correlated well with conventional ELISA methods and use of this technology provided several advantages over ELISA. PMID- 16309381 TI - Characterization of a cry4Ba-type gene of Bacillus thuringiensis israelensis and evidence of the synergistic larvicidal activity of its encoded protein with Cry2A delta-endotoxin of B. thuringiensis kurstaki on Culex pipiens (common house mosquito). AB - A newly isolated strain of Bacillus thuringiensis, named BUPM97, was identified as affiliated to the israelensis subspecies. This strain was selected for its insecticidal activity against larvae of several dipteran insects, such as the common house mosquito (Culex pipiens). A 5 kb EcoRI fragment, containing a cry4Ba type gene, named cry4BLB, was cloned from BUPM97. The sequencing of this gene revealed an open reading frame of 3411 bp encoding a protein of 1136 amino acid residues. Similarity analysis of both nucleotide and amino acid sequences revealed that cry4BLB presents several differences from the other previously described cry4Ba-type genes. Particular attention was paid to a two-amino-acid substitution located in domain III of the N-terminal moiety of this protein, which is very important for both toxicity and specificity of the toxin. The transfer of cry4BLB to an acrystalliferous B. thuringiensis kurstaki strain, HD1CryB, showed that it was expressed, resulting in the production of the typical parasporal crystal inclusions. On the other hand, its transfer to another B. thuringiensis kurstaki strain synthesizing Cry2A endotoxin known to be weakly toxic to the dipteran insect C. pipiens, resulted in the clear increase in the insecticidal activity of the transformed strain. Hence, in the present paper, we demonstrate for the first time that Cry2A and Cry4B act synergistically towards C. pipiens. PMID- 16309382 TI - Reconstruction of human hepatocyte glyoxylate metabolic pathways in stably transformed Chinese-hamster ovary cells. AB - Failure to detoxify the intermediary metabolite glyoxylate in human hepatocytes underlies the metabolic pathology of two potentially lethal hereditary calcium oxalate kidney stone diseases, PH (primary hyperoxaluria) types 1 and 2. In order to define more clearly the roles of enzymes involved in the metabolism of glyoxylate, we have established singly, doubly and triply transformed CHO (Chinese-hamster ovary) cell lines, expressing all combinations of normal human AGT (alanine:glyoxylate aminotransferase; the enzyme deficient in PH1), GR/HPR (glyoxylate/hydroxypyruvate reductase; the enzyme deficient in PH2), and GO (glycolate oxidase). We have embarked on the preliminary metabolic analysis of these transformants by studying the indirect toxicity of glycolate as a simple measure of the net intracellular production of glyoxylate. Our results show that glycolate is toxic only to those cells expressing GO and that this toxicity is diminished when AGT and/or GR/HPR are expressed in addition to GO. This finding indicates that we have been able to reconstruct the glycolate-->glyoxylate, glyoxylate-->glycine, and glyoxylate-->glycolate metabolic pathways, catalysed by GO, AGT, and GR/HPR respectively, in cells that do not normally express them. These results are compatible with the findings in PH1 and PH2, in which AGT and GR/HPR deficiencies lead to increased oxalate synthesis, due to the failure to detoxify its immediate precursor glyoxylate. These CHO cell transformants have a potential use as a cell-based bioassay for screening small molecules that stabilize AGT or GR/HPR and might have use in the treatment of PH1 or PH2. PMID- 16309383 TI - Vascular and fibrinolytic effects of intra-arterial tumour necrosis factor-alpha in patients with coronary heart disease. AB - Elevated plasma t-PA (tissue plasminogen activator) and serum CRP (C-reactive protein) concentrations are associated with an adverse cardiovascular risk. In the present study, we investigated whether acute local inflammation causes vascular dysfunction and influences t-PA release in patients with stable coronary heart disease. Serum CRP, plasma t-PA and PAI-1 (plasminogen activator inhibitor type 1) concentrations were determined in 95 patients with stable coronary heart disease. A representative subpopulation of 12 male patients received an intra brachial infusion of TNF-alpha (tumour necrosis factor-alpha) and saline placebo using a randomized double-blind cross-over study design. Forearm blood flow and plasma fibrinolytic and inflammatory variables were measured. Serum CRP concentrations correlated with plasma t-PA concentrations (r=0.37, P<0.001) and t PA/PAI-1 ratio (r=-0.21, P<0.05). Intra-arterial TNF-alpha caused a rise in t-PA concentrations (P<0.001) without affecting blood flow or PAI-1 concentrations. TNF-alpha pretreatment impaired acetylcholine- and sodium nitroprusside-induced vasodilatation (P<0.001 for both) whilst doubling bradykinin-induced t-PA release (P=0.006). In patients with stable coronary heart disease, plasma fibrinolytic factors correlate with a systemic inflammatory marker and local vascular inflammation directly impairs vasomotor function whilst enhancing endothelial t PA release. We suggest that the adverse prognosis associated with elevated plasma t-PA concentrations relates to the underlying causative association with vascular inflammation and injury. PMID- 16309384 TI - Golden age of drug discovery or dark age of missed chances? PMID- 16309385 TI - More cool news from marine bacteria. PMID- 16309386 TI - Bacterial degradation of xenobiotic compounds: evolution and distribution of novel enzyme activities. AB - Bacterial dehalogenases catalyse the cleavage of carbon-halogen bonds, which is a key step in aerobic mineralization pathways of many halogenated compounds that occur as environmental pollutants. There is a broad range of dehalogenases, which can be classified in different protein superfamilies and have fundamentally different catalytic mechanisms. Identical dehalogenases have repeatedly been detected in organisms that were isolated at different geographical locations, indicating that only a restricted number of sequences are used for a certain dehalogenation reaction in organohalogen-utilizing organisms. At the same time, massive random sequencing of environmental DNA, and microbial genome sequencing projects have shown that there is a large diversity of dehalogenase sequences that is not employed by known catabolic pathways. The corresponding proteins may have novel functions and selectivities that could be valuable for biotransformations in the future. Apparently, traditional enrichment and metagenome approaches explore different segments of sequence space. This is also observed with alkane hydroxylases, a category of proteins that can be detected on basis of conserved sequence motifs and for which a large number of sequences has been found in isolated bacterial cultures and genomic databases. It is likely that ongoing genetic adaptation, with the recruitment of silent sequences into functional catabolic routes and evolution of substrate range by mutations in structural genes, will further enhance the catabolic potential of bacteria toward synthetic organohalogens and ultimately contribute to cleansing the environment of these toxic and recalcitrant chemicals. PMID- 16309387 TI - Bacterial diversity of metagenomic and PCR libraries from the Delaware River. AB - To determine whether metagenomic libraries sample adequately the dominant bacteria in aquatic environments, we examined the phylogenetic make-up of a large insert metagenomic library constructed with bacterial DNA from the Delaware River, a polymerase chain reaction (PCR) library of 16S rRNA genes, and community structure determined by fluorescence in situ hybridization (FISH). The composition of the libraries and community structure determined by FISH differed for the major bacterial groups in the river, which included Actinobacteria, beta proteobacteria and Cytophaga-like bacteria. Beta-proteobacteria were underrepresented in the metagenomic library compared with the PCR library and FISH, while Cytophaga-like bacteria were more abundant in the metagenomic library than in the PCR library and in the actual community according to FISH. The Delaware River libraries contained bacteria belonging to several widespread freshwater clusters, including clusters of Polynucleobacter necessarius, Rhodoferax sp. Bal47 and LD28 beta-proteobacteria, the ACK-m1 and STA2-30 clusters of Actinobacteria, and the PRD01a001B Cytophaga-like bacteria cluster. Coverage of bacteria with > 97% sequence identity was 65% and 50% for the metagenomic and PCR libraries respectively. Rarefaction analysis of replicate PCR libraries and of a library constructed with re-conditioned amplicons indicated that heteroduplex formation did not substantially impact the composition of the PCR library. This study suggests that although it may miss some bacterial groups, the metagenomic approach can sample other groups (e.g. Cytophaga-like bacteria) that are potentially underrepresented by other culture-independent approaches. PMID- 16309388 TI - Aerobic anoxygenic photosynthesis genes and operons in uncultured bacteria in the Delaware River. AB - Photosynthesis genes and operons of aerobic anoxygenic photosynthetic (AAP) bacteria have been examined in a variety of marine habitats, but genomic information about freshwater AAP bacteria is lacking. The goal of this study was to examine photosynthesis genes of AAP bacteria in the Delaware River. In a fosmid library, we found two clones bearing photosynthesis gene clusters with unique gene content and organization. Both clones contained 37 open reading frames, with most of those genes encoding known AAP bacterial proteins. The genes in one fosmid were most closely related to those of AAP bacteria in the Rhodobacter genus. The genes of the other clone were related to those of freshwater beta-proteobacteria. Both clones contained the acsF gene, which is required for aerobic bacteriochlorophyll synthesis, suggesting that these bacteria are not anaerobes. The beta-proteobacterial fosmid has the puf operon B A-L-M-C and is the first example of an uncultured bacterium with this operon structure. The alpha-3-proteobacterial fosmid has a rare gene order (Q-B-A-L-M X), previously observed only in the Rhodobacter genus. Phylogenetic analyses of photosynthesis genes revealed a possible freshwater cluster of AAP beta proteobacteria. The data from both Delaware River clones suggest there are groups of freshwater or estuarine AAP bacteria distinct from those found in marine environments. PMID- 16309389 TI - Fishing for biodiversity: novel methanopterin-linked C transfer genes deduced from the Sargasso Sea metagenome. AB - The recently generated database of microbial genes from an oligotrophic environment populated by a calculated 1800 major phylotypes (the Sargasso Sea metagenome-SSM) presents a great source for expanding local databases of genes indicative of a specific function. In this article we analyse the SSM for the presence of methanopterin-linked C1 transfer genes that are signature for methylotrophy. We conclude that more than 10 phylotypes possessing genes of interest are present in this environment. The sequences representative of these major phylotypes do not appear to belong to any known microbial group capable of methanopterin-linked C1 transfer. Instead, these sequences separate from all known sequences on phylogenetic trees, pointing toward their affiliation with novel microbial phyla. These data imply a broader distribution of methanopterin linked functions in the microbial world than has been previously known. PMID- 16309390 TI - A wide host-range metagenomic library from a waste water treatment plant yields a novel alcohol/aldehyde dehydrogenase. AB - Using DNA obtained from the metagenome of an anaerobic digestor in a waste water treatment plant, we constructed a gene library cloned in the wide host-range cosmid pLAFR3. One cosmid enabled Rhizobium leguminosarum to grow on ethanol as sole carbon and energy source, this being due to the presence of a gene, termed adhEMeta. The AdhEMeta protein most closely resembles the AdhE alcohol dehydrogenase of Clostridium acetobutylicum, where it catalyses the formation of ethanol and butanol in a two-step reductive process. However, cloned adhEMeta did not confer ethanol utilization ability to Escherichia coli or to Pseudomonas aeruginosa, even though it was transcribed in both these hosts. Further, cell free extracts of E. coli and R. leguminosarum containing cloned adhEMeta had butanol and ethanol dehydrogenase activities when assayed in vitro. In contrast to the well-studied AdhE proteins of C. acetobutylicum and E. coli, the enzyme specified by adhEMeta is not inactivated by oxygen and it enables alcohol to be catabolized. Cloned adhEMeta did, however, confer one phenotype to E. coli. AdhE- mutants of E. coli fail to ferment glucose and introduction of adhEMeta restored the growth of such mutants when grown under fermentative conditions. These observations show that the use of wide host-range vectors enhances the efficacy with which metagenomic libraries can be screened for genes that confer novel functions. PMID- 16309391 TI - Screening a wide host-range, waste-water metagenomic library in tryptophan auxotrophs of Rhizobium leguminosarum and of Escherichia coli reveals different classes of cloned trp genes. AB - A metagenomic cosmid library was constructed, in which the insert DNA was derived from bacteria in a waste-water treatment plant and the vector was the wide host range cosmid pLAFR3. The library was screened for clones that could correct defined tryptophan auxotrophs of the alpha-proteobacterium Rhizobium leguminosarum and of Escherichia coli. A total of 26 different cosmids that corrected at least one trp mutant in one or both of these species were obtained. Several cosmids corrected the auxotrophy of one or more R. leguminosarum trp mutants, but not the corresponding mutants in E. coli. Conversely, one cosmid corrected trpA, B, C, D and E mutants of E. coli but none of the trp mutants of R. leguminosarum. Two of the Trp+ cosmids were examined in more detail. One contained a trp operon that resembled that of the pathogen Chlamydophila caviae, containing the unusual kynU gene, which specifies kynureninase. The other, whose trp genes functioned in R. leguminosarum but not in E. coli, contained trpDCFBA in an operon that is likely co-transcribed with five other genes, most of which had no known link with tryptophan synthesis. The sequences of these TRP proteins, and the products of nine other genes encoded by this cosmid, failed to affiliate them with any known bacterial lineage. For one metagenomic cosmid, lac reporter fusions confirmed that its cloned trp genes were transcribed in R. leguminosarum, but not in E. coli. Thus, rhizobia, with their many sigma-factors, may be well suited hosts for metagenomic libraries, cloned in wide host-range vectors. PMID- 16309392 TI - Insights into the genomes of archaea mediating the anaerobic oxidation of methane. AB - The anaerobic oxidation of methane is a globally significant process which is mediated by consortia of yet uncultivated methanotrophic archaea (ANME) and sulfate-reducing bacteria. In order to gain deeper insights into genome characteristics of the different ANME groups, large-insert genomic libraries were constructed using DNA extracted from a methanotrophic microbial mat growing in the anoxic part of the Black Sea, and from sediments above gas hydrates at the Hydrate Ridge off the coast of Oregon. Analysis of these fosmid libraries with respect to archaeal 16S rRNA gene diversity revealed a single ANME-1b ribotype for the Black Sea libraries, whereas the sequences derived from the Hydrate Ridge library phylogenetically affiliated with the ANME-2a, ANME-2c and ANME-3 group. Genome walking for ANME-1b resulted in a contiguous 155 kb composite genome fragment. The comparison of a set of four genomic fragments belonging to the different ANME groups revealed differences in the rRNA operon structure and the average G+C content, with the ANME-2c contig showing the highest divergence within the set. A detailed analysis of the ANME contigs with respect to genes putatively involved in the anaerobic oxidation of methane led to the identification of: (i) a putative N5,N10-methenyltetrahydromethanopterin cyclohydrolase gene, (ii) a gene cluster supposedly encoding a novel type of heterodisulfide reductase/dehydrogenase complex and (iii) a gene cluster putatively encoding a new type of CO dehydrogenase/acetyl-CoA synthase enzyme complex. PMID- 16309393 TI - Combined bromodeoxyuridine immunocapture and terminal-restriction fragment length polymorphism analysis highlights differences in the active soil bacterial metagenome due to Glomus mosseae inoculation or plant species. AB - High numbers of bacteria are associated with arbuscular mycorrhizal (AM) fungi, but their functions and in situ activities are largely unknown and most have never been characterized. The aim of the present study was to study the impact of Glomus mosseae inoculation and plant type on the active bacterial communities in soil by using a molecular approach, bromodeoxyuridine (BrdU) immunocapture in combination with terminal-restriction fragment length polymorphism (T-RFLP). This approach combined with sequence information from clone libraries, enabled the identification of actively growing populations, within the total bacterial community. Distinct differences in active bacterial community compositions were found according to G. mosseae inoculation, treatment with an antifungal compound (Benomyl) and plant type. The putative identities of the dominant bacterial species that were activated as a result of G. mosseae inoculation were found to be mostly uncultured bacteria and Paenibacillus species. These populations may represent novel bacterial groups that are able to influence the AM relationship and its subsequent effect on plant growth. PMID- 16309394 TI - Genetic and functional properties of uncultivated thermophilic crenarchaeotes from a subsurface gold mine as revealed by analysis of genome fragments. AB - Within a phylum Crenarchaeota, only some members of the hyperthermophilic class Thermoprotei, have been cultivated and characterized. In this study, we have constructed a metagenomic library from a microbial mat formation in a subsurface hot water stream of the Hishikari gold mine, Japan, and sequenced genome fragments of two different phylogroups of uncultivated thermophilic Crenarchaeota: (i) hot water crenarchaeotic group (HWCG) I (41.2 kb), and (ii) HWCG III (49.3 kb). The genome fragment of HWCG I contained a 16S rRNA gene, two tRNA genes and 35 genes encoding proteins but no 23S rRNA gene. Among the genes encoding proteins, several genes for putative aerobic-type carbon monoxide dehydrogenase represented a potential clue with regard to the yet unknown metabolism of HWCG I Archaea. The genome fragment of HWCG III contained a 16S/23S rRNA operon and 44 genes encoding proteins. In the 23S rRNA gene, we detected a homing-endonuclease encoding a group I intron similar to those detected in hyperthermophilic Crenarchaeota and Bacteria, as well as eukaryotic organelles. The reconstructed phylogenetic tree based on the 23S rRNA gene sequence reinforced the intermediate phylogenetic affiliation of HWCG III bridging the hyperthermophilic and non-thermophilic uncultivated Crenarchaeota. PMID- 16309395 TI - Novel genes for nitrite reductase and Amo-related proteins indicate a role of uncultivated mesophilic crenarchaeota in nitrogen cycling. AB - Mesophilic crenarchaeota are frequently found in terrestrial and marine habitats worldwide, but despite their considerable abundance the physiology of these as yet uncultivated archaea has remained unknown. From a 1.2 Gb large-insert environmental fosmid library of a calcareous grassland soil, a 43 kb genomic fragment was isolated with a ribosomal RNA that shows its affiliation to group 1.1b of crenarchaeota repeatedly found in soils. The insert encoded a homologue of a copper-containing nitrite reductase with an unusual C-terminus that encoded a potential amicyanin-like electron transfer domain as well as two proteins related to subunits of ammonia monooxygenases or particulate methane monooxygenases (AmoAB/PmoAB) respectively. Expression of nirK and the amoA-like gene was shown by reverse transcription polymerase chain reaction (PCR) analyses in soil samples, the latter being found at higher levels when the soil was incubated with ammonia (measured by quantitative PCR). Further variants of both genes were amplified from soil samples and were found in the environmental database from the Sargasso Sea plankton. Taken together, our findings suggest that mesophilic terrestrial and marine crenarchaeota might be capable of ammonia oxidation under aerobic and potentially also under anaerobic conditions. PMID- 16309396 TI - Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora. AB - A metagenome expression library of bulk DNA extracted from the rumen content of a dairy cow was established in a phage lambda vector and activity-based screening employed to explore the functional diversity of the microbial flora. Twenty-two clones specifying distinct hydrolytic activities (12 esterases, nine endo-beta 1,4-glucanases and one cyclodextrinase) were identified in the library and characterized. Sequence analysis of the retrieved enzymes revealed that eight (36%) were entirely new and formed deep-branched phylogenetic lineages with no close relatives among known ester- and glycosyl-hydrolases. Bioinformatic analyses of the hydrolase gene sequences, and the sequences and contexts of neighbouring genes, suggested tentative phylogenetic assignments of the rumen organisms producing the retrieved enzymes. The phylogenetic novelty of the hydrolases suggests that some of them may have potential for new applications in biocatalysis. PMID- 16309397 TI - Lateral gene transfer and phylogenetic assignment of environmental fosmid clones. AB - Metagenomic data, especially sequence data from large insert clones, are most useful when reasonable inferences about phylogenetic origins of inserts can be made. Often, clones that bear phylotypic markers (usually ribosomal RNA genes) are sought, but sometimes phylogenetic assignments have been based on the preponderance of blast hits obtained with predicted protein coding sequences (CDSs). Here we use a cloning method which greatly enriches for ribosomal RNA bearing fosmid clones to ask two questions: (i) how reliably can we judge the phylogenetic origin of a clone (that is, its RNA phylotype) from the sequences of its CDSs? and (ii) how much lateral gene transfer (LGT) do we see, as assessed by CDSs of different phylogenetic origins on the same fosmid? We sequenced 12 rRNA containing fosmid clones, obtained from libraries constructed using DNA isolated from Baltimore harbour sediments. Three of the clones are from bacterial candidate divisions for which no cultured representatives are available, and thus represent the first protein coding sequences from these major bacterial lineages. The amount of LGT was assessed by making phylogenetic trees of all the CDSs in the fosmid clones and comparing the phylogenetic position of the CDS to the rRNA phylotype. We find that the majority of CDSs in each fosmid, 57-96%, agree with their respective rRNA genes. However, we also find that a significant fraction of the CDSs in each fosmid, 7-44%, has been acquired by LGT. In several cases, we can infer co-transfer of functionally related genes, and generate hypotheses about mechanism and ecological significance of transfer. PMID- 16309398 TI - Putative novel photosynthetic reaction centre organizations in marine aerobic anoxygenic photosynthetic bacteria: insights from metagenomics and environmental genomics. AB - Photosynthetic core complexes of anoxygenic bacteria consist of reaction centres (RCs) surrounded by light-harvesting complexes (LHC). The structural proteins of the RC-LHC1 complex are encoded by the puf-operon. We find diverse operon organizations of puf-operons that reflect structural differences of the core complex in marine aerobic anoxygenic photosynthetic bacteria (AAnP). By analysis of environmental DNA records coming from AAnP bacteria we find several unknown proteins downstream to the pufM, which were assigned as novel PufX proteins. As all known pufX genes belong to Rhodobacter strains which carry out anaerobic photosynthesis, this may be the first observation of a PufX-containing RCs in aerobic anoxygenic photosynthetic bacteria. Phylogenetic analyses of PufM proteins from cultured as well as from uncultured bacteria show that PufM from operons containing putative novel pufX genes are grouped with Rhodobacter and not with Roseobacter strains. PMID- 16309399 TI - Acute effect of cerivastatin on cardiac regional ischemia in a rat model mimicking off-pump coronary surgery. AB - BACKGROUND: Although the off-pump technique is becoming widely used in coronary bypass surgery, there has been no experimental rat heart model of regional reversible ischemia. The aims of this study were to investigate the optimal duration of coronary occlusion for making reversible ischemia and to examine whether cerivastatin increases myocardial tolerance against prolonged coronary occlusion. METHODS: Study 1--Male Sprague-Dawley rats (350 to 450 g) underwent temporary occlusion of either left anterior descending artery (LAD; for 3, 5, 7.5, 10, 12.5, 15, or 20 min) or circumflex artery (CX; for 5, 10, or 15 min). Study 2--Rats were divided into two groups, control and cerivastatin groups, which had 0.1 mg/kg cerivastatin intravenously after anesthesia. LAD was occluded for 10, 15, or 20 minutes. In the both studies, hearts were stained to determine the area at risk (AR) and infarcted (IF) area 24 hours after reperfusion. RESULTS: In LAD occlusion, IF/AR increased in a time dependent manner: 4.5 +/- 3.2%, 9.7 +/- 5.2%, 17.2 +/- 3.0%, 16.8 +/- 2.7%, 23.9 +/- 9.5% (p < 0.01 vs. 3 min), 62.4 +/- 2.9% (p < 0.0001), and 63.4 +/- 2.9% (p < 0.0001) at 3, 5, 7.5, 10, 12.5, 15, and 20 min, respectively. Also in CX, IF/AR increased with time: 14.3 +/- 2.3%, 25.9 +/- 2.1%, and 40.9 +/- 6.2% (p < 0.001 vs. 5 min) at 5, 10, and 15 min, respectively. Cerivastatin significantly reduced IF/AR at 15 minutes (43.7 +/- 6.2%) and at 20 minutes (44.6 +/- 5.3%) compared to control (62.4 +/- 2.9% and 60.6 +/- 2.5%, respectively, p < 0.05). CONCLUSION: Cerivastatin increased myocardial tolerance after prolonged coronary occlusion over 10 minutes, which was considered to be the upper limit for creating a regional reversible ischemia in rats. PMID- 16309401 TI - Autologous blood transfusion in elective cardiac valve operations. AB - BACKGROUND AND AIM OF STUDY: The aim of this study was to detect any outcome differences between patients who donated autologous blood versus nondonors undergoing nonemergent cardiac valve surgery. Of further interest was whether autologous donors required less allogeneic blood products overall than patients who did not donate. METHODS: We conducted a nested case-control study in which data were collected prospectively on 225 variables. Cases underwent nonemergent, cardiac valve surgery and donated autologous blood products (n = 40). Controls also had nonemergent, cardiac valve surgery but did not donate autologous blood products (n = 120). Cases were matched to controls 1:3 on age (+/-3 years), gender, and New York Heart Association Functional Classification. We controlled for 12 potential confounding variables and examined 17 outcomes of interest. To generate the unadjusted risks of each outcome, chi-square and t-tests were performed comparing cases and controls to each outcome of interest. Then logistic regression analysis investigated the adjusted risk between cases and controls and for the outcomes of interest, each controlling for the potential confounding variables. RESULTS: There were no significant differences between the cases and controls for 11 of the 12 possible confounding variables. Controls had significantly more chronic obstructive pulmonary disorder. There were no significant differences between cases and controls for 13 of the 17 outcomes of interest. Autologous blood donors received more total packed red blood cells (PRBCs) (p = 0.0373) and more total fresh frozen plasma than controls (p = 0.0002). Fewer autologous blood donors required allogeneic packed red blood cell transfusion (p = 0.0134), and the total length of stay was shorter for autologous donors (p = 0.0782). CONCLUSION: Four of the 17 outcomes of interest were different for patients who donated autologous blood versus those who did not. Our experience demonstrated that elective cardiac valve surgery can safely reduce (by 18.3%) the need for allogeneic PRBCs by utilizing preoperative autologous blood donation. PMID- 16309402 TI - Aprotinin decreases ischemic damage during coronary revascularization. AB - BACKGROUND AND AIM: This study sought to determine whether the favorable anti inflammatory effects of aprotinin might limit ischemic damage during the revascularization of ischemic myocardium. METHODS: Twenty pigs underwent 90 minutes of coronary occlusion followed by 45 minutes of blood cardioplegic arrest and 180 minutes of reperfusion. Ten animals received a loading dose of aprotinin (40,000 kallikrein inhibiting units/kg) during the start of coronary occlusion followed by an infusion of 20,000 kallikrein inhibiting units/kg/hour. Ten other animals received no aprotinin. Summary statistics are expressed as the mean +/- standard error. RESULTS: The aprotinin-treated animals required less cardioversions for ventricular arrhythmias (1.0 +/- 0.7 vs. 3.6 +/- 0.6; p < 0.001), accumulated less lung water (1.0 +/- 0.2% change vs. 6.2 +/- 0.9% change; p = 0.038), had more complete coronary relaxation to bradykinin (34.1 +/- 5.9% change vs. 9.2 +/- 3.5% change; p = 0.01), and had reduced infarct size (area necrosis/area risk = 20 +/- 1.1% vs. 39 +/- 1.2%; p = 0.003). CONCLUSIONS: Aprotinin limits ischemic injury during acute coronary revascularization by decreasing ventricular arrhythmias and lung edema, preserving endothelial function, and minimizing myocardial necrosis. PMID- 16309403 TI - Aortic valve replacement for aortic insufficiency: valve type as a determinant of systolic strain recovery. AB - BACKGROUND AND AIM: Left ventricular (LV) 3D systolic strain decreases in absolute value postoperatively and does not recover in patients who undergo aortic valve replacement (AVR) for chronic aortic insufficiency (AI). We investigated whether choice of valve prosthesis (mechanical [St. Jude], bioprosthetic [bovine pericardial], Ross procedure) had a significant impact on strain recovery in this surgical population. METHODS: MRI with tissue-tagging was performed on 14 patients with chronic AI both before and 28 +/- 13 months after AVR. Average values of LV systolic strain and end-systolic stress (ESS) were computed from MRI data for the LV. Three types of prosthetic valve were examined (Ross procedure n = 4, bovine pericardial n = 5, and St. Jude n = 5). RESULTS: Overall, systolic strain, ESS, LV volumes, ejection fraction, and LV mass all changed significantly following AVR. Comparisons between individual valve types revealed no differences in any of these measurements. Patients who received a mechanical valve had a greater decrease in the absolute value of systolic strain following surgery compared to patients from the nonmechanical group (Ross procedure and bioprosthetic valve). Comparisons between the Ross group and the prosthetic group (St. Jude and bioprosthetic) produced no significant differences in strain, ESS, LV volume, and mass. CONCLUSIONS: These early results suggest that valve prosthetic type may be a factor in efforts to improve strain recovery after AVR for AI, although further investigation is warranted. MRI with tissue tagging may be a useful tool for comparing the impact of prosthetic valve choice on incompletely recovered systolic strain following AVR for chronic AI. PMID- 16309404 TI - Mid-term results of radial and mammary arteries as the conduits of choice for complete arterial revascularization in elective and nonelective coronary bypass surgery. AB - OBJECTIVE: The aim of this study is to evaluate the mid-results of the use of the radial artery alongside the internal mammary artery for complete arterial revascularization in elective and nonelective coronary bypass graft surgery. METHODS: All patients undergoing coronary artery surgery alone over a 3-year period with disease of more than one coronary artery were considered for complete arterial revascularization. Preoperatively, all patients had an Allen's test on the dominant arm and a cutoff point of 10 seconds was used. These patients were initially followed in the outpatient clinic after 6 weeks and then further followed up with the help of a mailed questionnaire. The incidence of recurrent angina and reangiogram was obtained and selected patients were further interviewed and examined in the outpatient department. RESULTS: Over a 3-year period, 291 patients underwent total arterial revascularization using the radial and internal mammary arteries alone in Y-graft configuration. The mean age of the study population was 62.4 +/- 8.8 years, with a male to female ratio of 221 to 70. Elective surgery was performed in 230 patients (79.4%), with nonelective procedures comprising a total of 61 patients (20.6%). The mean number of distal anastomoses was 2.9 +/- 0.9. There were four perioperative mortalities (1.37%) and 43 patients (14.7%) developed low cardiac output syndrome, requiring inotropes with or without intra-aortic balloon pump. Forty patients (13.7%) developed postoperative supraventricular arrhythmias. There was no incidence of hand ischemia or wound complications. After a mean follow-up period of 35.4 +/- 6.3 months of 220 patients (75%), there was one further death and 24 patients required readmissions for cardiac-related causes. Ten patients had reangiogram for angina of which one patient had a blocked radial artery graft and two patients underwent angioplasty to their native coronary arteries. The patients' angina score was currently 0.5 +/- 1.0 versus 2.6 +/- 1.4 preoperatively. CONCLUSION: Total arterial revascularization with the internal mammary and radial artery is associated with a low rate of perioperative complications and mortality and can be safely used in both elective and nonelective bypass graft surgery with excellent clinical results. PMID- 16309405 TI - Off-pump coronary artery grafting in patients with left main coronary artery disease. AB - BACKGROUND: Left main coronary artery disease (LMCD) is considered a relative contraindication to off-pump coronary artery bypass (OPCAB) grafting. This study evaluates the safety and feasibility of OPCAB in these patients compared to an on pump group (cardiopulmonary bypass, CPB) with LMCD. METHODS: Between January 2000 and July 2002, 95 patients with left main coronary artery stenosis >50% underwent coronary revascularization. Seventy-three unselected patients underwent OPCAB and 22 underwent revascularization using CPB. The techniques used for OPCAB included the use of deep traction sutures in the posterior pericardium and stabilizers to expose the distal coronary targets. Intraluminal coronary shunts were routinely used during construction of the anastomoses. Variables were analyzed using a Student's paired t-test with statistical significance defined as p < 0.05. RESULTS: The mean age in the OPCAB group was 59.9 years and the CPB group 61.8 years (p = 0.54). There were 56 males (77%) in the OPCAB and 18 (82%) in the CPB groups. Mean preoperative left ventricular ejection fraction (LVEF) was 40.3% in OPCAB and 47.3% in CPB (p = 0.015). Average number of grafts was 3.1 in OPCAB and 4.1 in CPB (p = 0.0038). There were no conversions to CPB in those patients initially chosen to undergo OPCAB. There were no early deaths in OPCAB. There was one death in CPB. Mean hospital length of stay was 6.9 days for OPCAB and 9.1 for CPB (p = 0.0159). CONCLUSIONS: Patients with LMCD can undergo OPCAB grafting safely and effectively despite reduced LVEF. LMCD should no longer be seen as a contraindication to perform OPCAB grafting. PMID- 16309406 TI - Chronic pseudoaneurysm of the aortic arch: a case report. AB - Pseudoaneurysm of the thoracic aorta is a rare condition and usually occurs following blunt trauma. It is almost fatal in the absence of prompt surgical treatment. We describe the case of a 56-year-old male suffering from intermittent non-massive hemoptysis, mild dysphagia, and atypical chest pain for 1 month who has no history of trauma. A saccular aneurysm at the aortic arch between left common carotid artery and left subclavian artery was diagnosed by magnetic resonance imaging. Intraoperatively, compression of surrounding structures including trachea and esophagus by the aneurysmal sac (6 x 8 cm in diameter) was seen. Pseudoaneurysm adherent to the upper lobe of the left lung was resected and entire aortic arch replacement with a prosthetic graft was performed. Postoperative course was uneventful and neither specific changes on histologic examination nor any evidence of infection could be detected. PMID- 16309407 TI - Acute myocardial infarction due to unruptured aneurysm of left sinus of valsalva with aortic valve regurgitation. AB - This report describes a 35-year-old woman with unruptured aneurysm of the left sinus of Valsalva presenting as non-ST elevation myocardial infarction due to the compression of the left coronary artery by aneurysm. Cardiac multislice CT and angiogram revealed a large aneurysm of the left sinus of Valsalva compressing the left main coronary artery. Surgical repair was performed by closing the entrance of the aneurysm and aortic valve replacement. Postoperative coronary flow was restored and thus anginal symptom disappeared. PMID- 16309408 TI - Reoperation for calcific constrictive pericarditis 43 years after pericardiectomy. AB - The case of calcific pericarditis in a 63-year-old male with a 1-year history of progressive right heart failure 43 years after pericardiectomy in 1960 is reported. The recurrent constriction was caused by the calcification of the residual posteroinferior pericardium and the impressing sternum of a pectus excavatum encasing the heart like a skull. The patient underwent total pericardiectomy of his residual pericardium through median sternotomy using cardiopulmonary bypass resulting in complete relief of his symptoms. PMID- 16309409 TI - Spontaneous resolution of type a aortic dissection. AB - We present the case of a middle-aged man who presented with type A aortic dissection, which was resolved spontaneously over 5 weeks time. We also discuss the possible mechanism and the role of conservative management in very selected group of patients. PMID- 16309410 TI - Coronary artery bypass grafting in a patient with Glanzmann's thrombasthenia. AB - Glanzmann's thrombasthenia is a platelet aggregation disorder resulting from a functional loss of platelet membrane glycoprotein IIb-IIIa. First described by Dr. Glanzmann in 1918, the disorder is characterized clinically by mucocutaneous bleeding and physiologically by absent platelet aggregation to collagen, epinephrine, and adenosine diphosphate stimulation. While there are multiple reports of patients with Glanzmann's thrombasthenia undergoing surgery, to our knowledge there has been no report of a patient with Glanzmann's undergoing coronary artery bypass grafting. We present the first such report of a patient who successfully underwent operative coronary artery revascularization, and offer suggestions for future management of these patients. PMID- 16309411 TI - Laparoscopic cholecystectomy and open-heart surgery at the same time. AB - BACKGROUND: Abdominal procedures in patients with coronary artery disease or severe valvular disease have high risk of mortality and morbidity. AIMS: In order to prevent bile peritonitis after cardiac surgery, to reduce the hospitalization course, and to decrease the mortality and morbidity after the surgery, laparoscopic cholecystectomy and open-heart surgery were performed at the same time. METHODS: Laparoscopic cholecystectomy and open-heart surgery were performed on two patients who had been referred to Cardiothoracic Ward of Modarres Hospital, Tehran, Iran. RESULTS: Simultaneous classic laparoscopic cholecystectomy was successfully performed on two different open-heart patients, one with coronary artery disease for coronary artery bypass surgery and the other with severe mitral valve disorder for mitral valve replacement. CONCLUSIONS: Abdominal procedures are suggested to be done with cardiac surgery at the same time with laparoscopic technique to reduce mortality and morbidity in these patients. PMID- 16309412 TI - Delirium in the postoperative cardiac patient: a review. AB - BACKGROUND AND AIM OF REVIEW: Cardiac surgery is increasingly common and relatively safe, but there are frequent reports of neuropsychiatric sequelae occurring in the postoperative period. One of the most common neuropsychiatric presentations of cardiac surgery is delirium, also called postcardiotomy delirium (PCD). Despite the vast numbers of cardiac surgeries performed today, there is a paucity of data on risk factors and management options of PCD available to the clinician. This review aims to summarize available information, increase clinicians' awareness of PCD and suggest effective management of this illness. METHODS: Our literature search was completed using the databases Medline and CINAHL; it was limited to human and English language studies from 1964 to the present. Search terms included "delirium," "agitation," "postoperative," "cardiac," "neuropsychiatric," "neuroleptics," "psychosis," "surgery," "treatment," "postcardiotomy," and "pharmacotherapy." RESULTS: Our review of the literature revealed several risk factors for PCD, as well as various options for its pharmacological management. CONCLUSIONS: A multifactorial model should be applied when considering risk stratification for and prevention of delirium postoperatively. Pharmacologically, conventional antipsychotic agents, such as haloperidol, have long been used to manage delirium. In light of haloperidol's side effects, particularly those applicable to the cardiac patient, further research is required into the role of second generation antipsychotics. These agents are common in clinical use, and may be the preferred medications. PMID- 16309414 TI - Pioneering contributions of Alexander A. Vishnevsky and his team to cardiac surgery. AB - In the dawn of cardiac surgery in the 1940s to 1950s, Alexander A. Vishnevsky assembled together a fine team of surgeons and conducted intense surgical research that resulted in clinical application of modified Blalock-Taussig shunt, cavo-pulmonary shunt, and paved the way to clinical orthotopic heart and lung transplantation and coronary bypass surgery in Russia. The team of surgeons lead by Vishnevsky also performed the world's first cavo-pulmonary shunts and experimental orthotopic heart and lung transplantation. Herein, we describe the pioneering contributions of Vishnevsky and his team to cardiac surgery. PMID- 16309415 TI - Efficacy of Blake drains for mediastinal and pleural drainage following cardiac operations. AB - BACKGROUND: Mediastinal and pleural drainage following cardiac operations has traditionally been achieved with large bore, semirigid chest tubes. The purpose of this study was to evaluate the safety and efficacy of drainage by means of small, soft, and flexible 19 F Blake drains. METHODS: This is a review of all patients who underwent heart surgery over a 3-year period at a single institution. Chest tubes and Blake drains were removed on postoperative day 1 to 5 depending on patient's condition, amount of drainage, and surgeon's preference. The criteria for drain removal did not vary with type of drain. RESULTS: There was no significant difference in the amount of drainage between both groups. Postoperative mediastinal exploration occurred in 3.47% of patients (12/346) in the chest tube group and in 2.08% of patients (8/385) in the Blake group (p = 0.27). Significant pleural effusions requiring a subsequent drainage procedure occurred in 9.54% of patients (33/346) in the chest tube group and in 9.87% of patients (38/385) in the Blake group. CONCLUSIONS: No significant differences were noted in the number of mediastinal explorations in patients drained with conventional chest tubes as compared to Blake drains during cardiac operations. Though not statistically significant, there may actually be an advantage of Blake drains over conventional chest tubes in this regard. There was also no significant difference in the incidence of postoperative pleural effusions. Blake drains appear to be at least as effective and safe as conventional chest tubes in draining the mediastinum and pleural spaces following cardiac surgery. PMID- 16309416 TI - Onlay patch for complete intercostal artery preservation during thoracic and thoracoabdominal aortic aneurysm repair. AB - BACKGROUND: Replacement of the thoracic aorta is associated with a potential for interruption of the blood supply to the spinal cord leading to paralysis. Techniques have been proposed to ameliorate this risk, including reimplantation of the intercostal arteries. We present a technique to simplify this approach. METHODS: In patients with extensive aneurysmal disease, a standard approach to the thoracic aorta via a left thoracotomy is performed. The patient is placed on full cardiopulmonary bypass with moderate hypothermia. The proximal anastomosis is performed unclamped with circulatory arrest. After completing the proximal and distal anastomosis separately, an onlay patch of Dacron is created and placed along the entire back wall of the aorta to cover all of the intercostal arteries. A side-to-side anastomosis between the patch and the distal graft is then performed and subsequently the anastomosis tested by retrograde flow. RESULTS: The technique allows direct inspection of all suture lines, tested to an adequate pressure, so that postoperative bleeding is minimal. CONCLUSION: Techniques to lessen the risk of paraplegia associated with aortic surgery include reimplantation of the intercostal arteries. This technique allows each suture line to be tested and easily visually inspected prior to closure of the chest. PMID- 16309418 TI - Free right internal thoracic artery in a "horseshoe" configuration: a new technical approach for "in situ" conduit lengthening. AB - Severe chronic obstructive pulmonary disease with large lung volumes may prevent both the "in situ" internal thoracic arteries to reach coronary anastomoses sites. We present a method to revascularize the left antero-lateral myocardial wall using the right internal thoracic artery as a "free graft" anastomosed side to end to the "in situ" left internal thoracic artery, in a "horseshoe" fashion. The two ends of the "free graft" were anastomosed to the left anterior descending coronary artery and the second obtuse marginal branch, respectively. This method was successfully used in a 74-year-old patient with severe chronic obstructive pulmonary disease. PMID- 16309420 TI - Off-pump axillo-coronary artery bypass grafting with porcelain aorta. AB - Coronary artery bypass grafting (CABG) in patients with severely diseased ascending aorta has been associated with high risk for cerebral vascular accidents due to atheromatous embolism. In this situation, aortic no-touch techniques are widely employed as most important surgical strategy to prevent these complications. A case of 75-year-old man with effort angina associated with porcelain ascending aorta was reported here. He successfully underwent off-pump axillo-coronary bypass grafting with saphenous vein graft and has remained uneventful during his follow-up period. The off-pump axillo-coronary artery bypass grafting seemed to be an appropriate procedure for coronary revascularization with severely diseased ascending aorta. PMID- 16309421 TI - Severe truncal valve stenosis: diagnosis and management. AB - Severe truncal valve stenosis, due to dysplastic valve, may be found in a small number of patients with truncus arteriosus. Its presence complicates the surgical management of this anomaly due to lack of optimal therapeutic options and associated ventricular dysfunction. We report a case of truncal root replacement using a homograft for the management of such condition, and review the literature related to its management and outcome. PMID- 16309422 TI - Anti-HBV effects of CpG oligodeoxynucleotide-activated peripheral blood mononuclear cells from patients with chronic hepatitis B. AB - Unmethylated CpG dinucleotides in bacterial DNA or synthetic oligodeoxynucleotides containing immunostimulatory CpG motifs (CpG ODN) are known as a potent Th1-like immune enhancer in vertebrates. Chronic hepatitis B is the immunocompromising condition. We therefore investigated the effects of CpG ODN on cultured cells from chronic hepatitis B patients and healthy controls. The inhibitory effects of CpG ODN on hepatitis B virus (HBV) were also studied. The secretion of IFN-alpha by CpG ODN-activated peripheral blood mononuclear cells (PBMCs) from chronic hepatitis B patients and healthy controls was significantly increased when compared with PBMCs alone or GpC ODN-stimulated PBMCs. After activation with CpG ODN, the IFN-alpha secretion by chronically HBV-infected patient PBMCs is less than that by healthy control PBMCs. Treatment of HepG2 2.2.15 cells with culture supernatants of PBMCs activated by CpG ODN can significantly suppress the secretion of HBsAg, HBeAg and HBV DNA as compared with that of PBMCs without CpG ODN activation under the same conditions. No inhibitory effect on the replication of HBV was found for CpG ODN treatment alone. Our results indicated that CpG ODN could efficiently enhance the immune response of chronic hepatitis B patients. Moreover, the CpG ODN-activated PBMCs from chronic hepatitis B patients were able to significantly inhibit HBV replication in vitro, suggesting that CpG ODN may be a potential immunoregulator against HBV infection in the future. PMID- 16309423 TI - Kinetic bactericidal activity of telithromycin, azithromycin and clarithromycin against respiratory pathogens. AB - The present study assessed the comparative in vitro killing kinetics of telithromycin, azithromycin and clarithromycin. Minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) were determined against Streptococcus pneumoniae, beta-haemolytic streptococci, Haemophilus influenzae and Moraxella catarrhalis strains characterized by different susceptibilities to beta-lactams and macrolides. For each bacterial species, representative strains were chosen for time-kill studies. Telithromycin showed high activity against all the tested strains with MIC ranging from < or = 0.004 to 0.5 mg/L for streptococci, from 0.008 to 8 mg/L for H. influenzae, and from 0.008 to 0.5 mg/L for M. catarrhalis. In time-kill studies, telithromycin showed an overall superior bactericidal activity in respect to macrolides, particularly against resistant strains. In conclusion, telithromycin proved to possess bactericidal activity against a wide range of respiratory pathogens, including strains resistant to common macrolides. PMID- 16309424 TI - Prevalence of Legionella pneumophila serogroup 1 in water distribution systems in Izmir province of Turkey. AB - Legionella pneumophila serogroup 1 occurrence has been investigated in 168 hot water samples from 24 hotels, situated in 6 counties in Izmir province of Turkey, from 15 June to 30 September of the year 2000. Sampling was carried out at 15-day intervals and seven samples were taken from each of the hotels' hot water reservoirs and hot water networks. The samples were (1 L) concentrated using polycarbonate filters (mesh size 0.22 microm). Isolation was achieved using selective medium, GVPC agar. The samples were concentrated by membrane filtration, divided into three portions and cultured without pretreatment, after acid treatment, and after heat treatment, on GVPC agar. One hundred and ten isolates were identified as L.pneumophila sg 1 using the Legionella Latex Test (Oxoid). Arbitrarily primed PCR (AP PCR) was employed to assess the clonal relationship between Legionella pneumophila sg 1 isolates from the hot water samples of the hotels. Three genotypes of L. pneumophila sg 1 isolates were identified. With a high prevalence of type A, 22 hotels were found to be colonized with L. pneumophila serogroup 1, while only 2 were free from the bacteria. PMID- 16309425 TI - Evidence for a reserpine-affected mechanism of resistance to tetracycline in Neisseria gonorrhoeae. AB - The presence of a reserpine-affected mechanism of tetracycline resistance was investigated in 17 Neisseria gonorrhoeae clinical isolates. To establish this fact the MIC of tetracycline in the presence and absence of reserpine was determined, and, in addition, mechanisms of tetracycline resistance were analyzed by PCR. The results showed that reserpine affects the MIC of tetracycline at least 4-fold in all isolates, including those containing the tetM gene. An inhibitory effect of reserpine against the MtrCDE efflux system was ruled out by using strains either with an inactive or with an unrepressed MtrCDE system. The results suggest the presence of a constitutive system of resistance to tetracycline, by a possible efflux pump, which may be inhibited by reserpine. Further studies are required to determine the exact nature of the action of reserpine on the MIC of tetracycline. PMID- 16309426 TI - The white pulp in the setting of the septic spleen caused by different bacteria: a comparative morphometric study. AB - In the past little attention has been paid to histomorphologic changes accompanying the phenomenon of the septic spleen, thus indirectly reinforcing the old axiom that the spleen is an organ of mystery. It is especially noteworthy that the relationship between different causative bacteria and histopathologic abnormalities of the white pulp has not been investigated. In this study morphometric analysis was performed on the white pulp of 30 spleens obtained at autopsy from individuals with premortal sepsis. A strictly defined age- and sex matched control group was analyzed for statistical comparison. Our findings demonstrate a significant depletion of B- and T-areas in the septic spleen, accompanied by a significant tendency towards reactive germinal center hyperplasia regardless of the type of bacteria responsible. However, depletion of splenic B-areas was shown to be significantly pronounced in the setting of premortal enterococcemia in comparison with a panel of gram-negative flagellated bacteria. It is felt that certain bacterial virulence factors (e.g. flagellation and/or structural components of the cell wall) might be pathogenetically involved in the observed changes, reflecting a partially different activation of splenic lymphocytes in the setting of the septic spleen. PMID- 16309427 TI - ERBB2 kinase domain mutation in a gastric cancer metastasis. AB - ERBB2 is a member of the epidermal growth factor receptor (EGFR) family. Recent studies revealed that the kinase domain of the ERBB2 gene was mutated in human cancers, including gastric cancer. Despite the importance of cancer metastasis in the pathogenesis of cancers, data on the ERBB2 kinase domain mutation in cancer metastasis are lacking. In this study, to explore the possibility that ERBB2 mutation is involved in the metastasis mechanism, we analyzed the kinase domain of ERBB2 for the detection of somatic mutations in 58 gastric adenocarcinomas with lymph node metastasis. We found one ERBB2 mutation, which was detected in the lymph node metastasis, but not in the primary tumor of the same patient. The ERBB2 mutation was a missense mutation which substituted an amino acid in exon 21 (V832I). We simultaneously analyzed the somatic mutations of EGFR, K-RAS, PIK3CA and BRAF genes in the sample with the ERBB2 mutation, and found that this metastatic carcinoma did not harbor any of the mutations. Our data suggest that ERBB2 kinase domain mutation occasionally occurs in metastatic gastric carcinoma and might play a role in the metastatic process of some gastric carcinomas. PMID- 16309428 TI - Detection of methicillin resistance in coagulase-negative staphylococci by cefoxitin disc diffusion and oxacillin Etest. A study of consecutive bacteraemia isolates. AB - Coagulase-negative staphylococci (CoNS) are a significant cause of nosocomial bacteraemia and their susceptibility to beta-lactamase-stabile penicillins is unpredictable. To ensure appropriate antibiotic therapy reliable methods for detection of methicillin resistance (MR) are needed. The objectives of this study were to determine the frequency of MR in a set of CoNS from cases of monomicrobial bacteraemia and to evaluate two phenotypic assays for detection of MR, the 10 microg cefoxitin disk test on Iso-Sensitest agar using a semiconfluent inoculum and the oxacillin Etest. MR was determined by a commercial genomic mecA assay. Of 110 CoNS, 75 were mecA positive and 35 mecA negative. Using interpretive zone diameters R < 22 mm and S > or = 27 mm, the cefoxitin disk test had a sensitivity and specificity of 100%. A correct prediction was obtained for 86 isolates, while 23 were indeterminate (> or = 22 mm; <27 mm). Using CLSI's guidelines, sensitivity and specificity of the oxacillin Etest were 100% and 80%, respectively. A correct prediction was obtained for 102 isolates, while 7 mecA negative isolates were classified as resistant. Thus, the cefoxitin disk test and the oxacillin Etest performed with high accuracy and both seem to be suitable for routine use. PMID- 16309429 TI - Effect of different ERK2 protein localizations on prognosis of patients with invasive breast carcinoma. AB - Mitogen-activated protein kinase (MAP kinase) pathways represent a cascade of phosphorylation events, including three pivotal kinases, Raf, MEK and ERK1/2, which have been implicated in the pathogenesis of cancer. We examined 151 cases of invasive breast carcinoma by immunohistochemistry and compared the ERK2 expression with clinicopathological parameters, MMP-11 immunoexpression and patients' survival. ERK2 immunoexpression was detected in the cytoplasm and nucleus of cancer cells in 37.7% and 19.2% of cases, respectively. Nuclear ERK2 was inversely correlated with ER (p = 0.039), whereas cytoplasmic ERK2 was positively correlated with MMP-11 in fibroblasts (p = 0.032) and more often expressed in lobular than ductal carcinomas (p = 0.026). Nuclear ERK2 expression was found to be an independent prognostic factor of shortened overall survival of patients (p = 0.040), while cytoplasmic ERK2 had an independent, favorable effect on both disease-free and overall survival (p < 0.0001 and p = 0.002, respectively). These findings suggest that the different subcellular localizations of ERK2 seem to be related to different, possibly contradictory, effects on patient survival. PMID- 16309430 TI - Juvenile kyphosis in pigs. A spontaneous model of Scheuermann's kyphosis. AB - The development of kyphotic lesions in pigs with different weights from herds with a high frequency of the lesion was analysed pathologically, radiologically, and for alkaline phosphatase, calcium, magnesium, and phosphate in blood samples. The development of kyphosis was caused by the formation of ventral hemivertebrae due to the absence of ventral vertebral epiphyseal centres of ossification. Within the ventral parts of affected vertebral epiphyses, the retained chondroid tissue was dysplastic and the contents of cartilage canals with vessels often clotted with fibrin were reduced. As lesions such as fractures and inflammation that may be a secondary cause of kyphosis in pigs were absent and all blood parameters were within normal range, secondary causes of the condition, including metabolic disturbances, were excluded. It can be seen that the present type of porcine kyphosis does not develop until later in life as the ossification centres within the epiphyses of vertebrae have a retarded appearance. As the present porcine type of kyphosis pathomorphologically is comparable with Scheuermann's kyphosis in man, it constitutes a spontaneous model for this common cause of structural kyphosis of the thoracic or thoracolumbar spine. PMID- 16309431 TI - White variants of Trichophyton violaceum isolated in Ethiopia. AB - Certain dermatophytes are geographically restricted and endemic in particular parts of the world, while other species may have a sporadic but worldwide distribution. Trichophyton violaceum is one of the most common dermatophytes causing tinea capitis, and is the predominant cause of tinea in Africa, South America and the Indian subcontinent. Among 1187 dermatophyte isolates collected from Ethiopian patients with various types of tinea, 32 isolates had uncharacteristic phenotypic features. Based on conventional methods complemented by sequence analysis of the rDNA ITS2 region, these isolates were identified as white variants of T. violaceum. This is the first time that white isolates of T. violaceum have been identified in Ethiopia. PMID- 16309432 TI - Serous papillary adenocarcinoma and adult granulosa cell tumor in the same ovary. An unusual case. AB - Surface epithelial-stromal cell tumors are the most common neoplasms of the ovary but occurrence of a serous adenocarcinoma and an adult granulosa cell tumor in the same ovary is an unusual incident. In the present case report we describe this very uncommon occurrence in the ovary of a 50-year-old woman. The patient suffered abdominal distention and was referred to the state hospital where a 5x3 cm multilocular cystic lesion was observed on abdominal CT. Total abdominal hysterectomy with salpingo-oophorectomy and omentectomy was performed. Microscopy revealed an adult granulosa cell tumor and a serous papillary adenocarcinoma in the left ovary. Immunohistochemical staining with inhibin alpha and pancytokeratin confirmed the diagnosis. PMID- 16309433 TI - Clear cell sarcoma of the small bowel: a potential pitfall. Case report. AB - Clear cell sarcoma (soft-part melanoma) is a very rare entity with a distinctive histopathologic and molecular profile. Herein, we present the sixth reported case of a primary gastrointestinal clear cell sarcoma discovered in a 21-year-old woman. The patient underwent numerous tests prior to the diagnosis of her small bowel pathology, including the use of capsule endoscopy, which allowed for visualization and final localization of the tumour. Additionally, we discuss this rare type of sarcoma that affects young adults and has a poor prognosis characterized by the balanced chromosomal translocation t(12;22)(q13;q12) with special emphasis on the necessity for pathologists to be able to distinguish it from melanoma -- potentially a major pitfall in diagnosis. PMID- 16309434 TI - Cystic dysplasia of the rete testis. Case report. AB - Cystic dysplasia of the rete testis (CDRT) is a very rare cause of a paediatric scrotal mass often associated with renal and other genitourinary tract anomalies. These complex malformations are probably due to a developmental defect of the mesometanephric system during embryogenesis. A case of asymptomatic scrotal swelling in a 4-year-old boy is presented. Ultrasonography, showed a cystic lesion of the left testis associated with absence of the left kidney. Orchiectomy was performed because of extensive gonad involvement. Pathologic examination revealed multiple, anastomosing, irregular cystic spaces of varying sizes and shapes predominantly located in the region of the rete testis. The cysts had spread irregularly, displacing the testicular parenchyma, which was subsequently compressed under the tunica albuginea. Preoperative diagnosis of CDRT is easy if age, precise localisation, characteristic ultrasonographic features and other genitourinary malformations are considered. Other paediatric cystic lesions should be included in the differential diagnosis. It is possible to cure CDRT by orchiectomy or by conservative treatment. Nowadays the later option is preferred, but diagnosis of CDRT must be precisely established and followed by careful monitoring. PMID- 16309435 TI - Second modernity as a research agenda: theoretical and empirical explorations in the 'meta-change' of modern society. AB - In this article we are reformulating the theory of reflexive modernization as an empirical research programme and summarize some of the most recent findings which have been produced by a research consortium in Munich (integrating four universities, funded by the German Research Society (DFG)). On this basis we reject the idea that Western societies at the beginning of the twenty-first century move from the modern to the post-modern. We argue that there has been no clear break with the basic principles of modernity but rather a transformation of basic institutions of modernity such as the nation-state and the nuclear family. We would suggest, therefore, that what we are witnessing is a second modernity. Finally, we reform the theory of reflexive modernization in reaction to three uttered objections. PMID- 16309436 TI - Global generations: social change in the twentieth century. AB - The concept of generation within sociology has until recently been a marginal area of interest. However, various demographic, cultural and intellectual developments have re-awakened an interest in generations that started with the classic essay by Karl Mannheim. To date, the sociological literature has generally conceptualized generations as nationally bounded entities. In this paper we suggest that the sociology of generations should develop the concept of global generations. This conceptual enhancement is important because the growth of global communications technology has enabled traumatic events, in an unparalleled way, to be experienced globally. The late nineteenth and early twentieth century was the era of international generations, united through print media, and the mid-twentieth century saw the emergence of transnational generations, facilitated by new broadcast communications. However, the latter part of the twentieth century is the period of global generations, defined by electronic communications technology, which is characterized, uniquely, by increasing interactivity. The 1960s generation was the first global generation, the emergence of which had world-wide consequences; today with major developments in new electronic communications, there is even more potential for the emergence of global generations that can communicate across national boundaries and through time. If in the past historical traumas combined with available opportunities to create national generations, now globally experienced traumas, facilitated by new media technologies, have the potential for creating global generational consciousness. The media have become increasingly implicated in the formation of generational movements. Because we are talking about generations in the making rather than an historical generation, this article is necessarily speculative; it aims to provoke discussion and establish a new research agenda for work on generations. PMID- 16309437 TI - Understanding generations: political economy and culture in an ageing society. AB - Sociological understanding of generations can be enhanced by avoiding defining them rigidly as chronological cohorts but rather linking people's accounts of their generational experience with an historically informed political economy. It then becomes possible, for example, to understand the complexity of generational politics. This paper uses data on the 'War Generation' taken from the Exeter Politics of Old Age project to link an empirically based political economy of generational inequality with a cultural sociology of generations. The 'War Generation' recognizes itself and is referred to by others in terms of a common identity. It is also an historical generation; its values, attitudes and, above all, sense of national solidarity and mutual obligation were forged in the direct experience of war. But it is also divided by divergent economic interests in property and pension rights based on the historical experience of the life course by successive groups and this segmentation can be observed in political action. The political culture of the War Generation manifests both continuity and change. Understanding these dynamics requires listening to people constructing their worlds, understanding their full range of historical experiences, and analysing the conditions for their conflicts and their cohesion. PMID- 16309438 TI - The new gender essentialism--domestic and family 'choices' and their relation to attitudes. AB - This paper critically examines two strands within contemporary gender essentialism--that is, the argument that men and women are fundamentally different and that it is this 'difference' that explains the continuing social and material differences between the sexes. The first strand we examine is Hakim's 'preference theory', which has argued that persisting sex differences in employment patterns are an outcome of the 'choices' made by different 'types' of women. We next examine the claims of populist conservative feminism, that has argued that women (and men) in partnerships will be happier if they adopt a gender role traditionalism in the domestic sphere. Our empirical findings suggest that neither of these theoretical explanations are supported by our data, which is derived from the samples of six countries participating in the International Social Survey Programme Family 2002 module. PMID- 16309439 TI - Three current issues in ethnic and racial studies. AB - Prevailing orthodoxies in British studies of ethnic and racial relations have been challenged, most recently by perspectives developed within cultural studies. The issues that divide the contending parties have still to be clarified. Three of them appear to be: (i) problem definition; (ii) problem selection; and (iii) conceptual obsolescence. PMID- 16309440 TI - Between football and martyrdom: the bi-focal localism of an Arab-Palestinian town in Israel. AB - Studies of the relationship between nationalism and localism have brought evidence that these orientations might maintain either relations of opposition or congruence. By conceiving of localism mainly as a strategy, this paper argues that localism can be used alternately as an anchor of nationalist narrative or as an alternative to nationalism by the exact same community. This argument is illustrated by the case of Sakhnin, an Arab-Palestinian town in Israel. Local pride in Sakhnin has developed around two separate foci: a nationalist heroic narrative of martyrdom and the success of the local football team. These two foci developed in complete isolation, since the first is embedded in a Palestinian nationalist narrative while the latter is oriented toward the Israeli Jewish public. An ethnographic study follows the construction of these separate spheres and a survey conducted among 174 men in the town confirms that involvement in the football sphere correlates with both local pride and integrative orientations. PMID- 16309456 TI - Citrobacter rodentium of mice and man. AB - The major classes of enteric bacteria harbour a conserved core genomic structure, common to both commensal and pathogenic strains, that is most likely optimized to a life style involving colonization of the host intestine and transmission via the environment. In pathogenic bacteria this core genome framework is decorated with novel genetic islands that are often associated with adaptive phenotypes such as virulence. This classical genome organization is well illustrated by a group of extracellular enteric pathogens, which includes enteropathogenic Escherichia coli (EPEC), enterohaemorrhagic E. coli (EHEC) and Citrobacter rodentium, all of which use attaching and effacing (A/E) lesion formation as a major mechanism of tissue targeting and infection. Both EHEC and EPEC are poorly pathogenic in mice but infect humans and domestic animals. In contrast, C. rodentium is a natural mouse pathogen that is related to E. coli, hence providing an excellent in vivo model for A/E lesion forming pathogens. C. rodentium also provides a model of infections that are mainly restricted to the lumen of the intestine. The mechanism's by which the immune system deals with such infections has become a topic of great interest in recent years. Here we review the literature of C. rodentium from its emergence in the mid-1960s to the most contemporary reports of colonization, pathogenesis, transmission and immunity. PMID- 16309457 TI - Immunologic memory in cutaneous leishmaniasis. AB - Leishmania major infections induce solid immunity to reinfection. Experimental studies in mice indicate that the CD4+ T cells responsible for this immunity include two populations: parasite-dependent T effector cells and parasite independent central memory T (Tcm) cells. While there currently is no vaccine for leishmaniasis, the existence of a long-lived population of Tcm cells that does not require the continued presence of live parasites suggests that a vaccine that expands these cells might be efficacious. PMID- 16309458 TI - Recent insights into the mechanisms of Chlamydia entry. AB - Chlamydia are widespread bacteria that grow in human and animal cells. They enter their host cell, establish an intracellular environment favourable for their multiplication and finally exit the host cell. A combination of host cell factors and of bacterial proteins contribute to pathogen entry. Recent advances have shed new light on the entry mechanism, following attachment. Here we review recent data concerning endocytosis, host cell signalling, proteins secreted by the bacteria, the actin cytoskeleton in entry and the involvement of small GTPases. PMID- 16309459 TI - The Steinernema carpocapsae intestinal vesicle contains a subcellular structure with which Xenorhabdus nematophila associates during colonization initiation. AB - Steinernema carpocapsae infective juvenile (IJ) nematodes are intestinally colonized by mutualistic Xenorhabdus nematophila bacteria. During IJ development, a small number of ingested X. nematophila cells initiate colonization in an anterior region of the intestine termed the vesicle and subsequently multiply within this host niche. We hypothesize that efficient colonization of a high percentage of S. carpocapsae individuals (typically>85%) is facilitated by bacterial adherence to a site(s) in the nematode intestine. We provide evidence that the adherence site is a structure in the lumen of the IJ vesicle that we have termed the intravesicular structure (IVS). The IVS is an untethered cluster of anucleate spherical bodies that co-localizes with colonizing X. nematophila cells, but does not require X. nematophila for its formation. Colocalization with the IVS is readily apparent in IJs colonized by X. nematophila mutants that initiate intestinal colonization but fail to proliferate normally, suggesting that bacterial-IVS interaction occurs early in the colonization process. Treatment with insect haemolymph induces anal release of X. nematophila from colonized IJs and induces release of the IVS from uncolonized S. carpocapsae IJs. Released IVS were probed with several carbohydrate-specific lectins. One lectin, wheat-germ agglutinin, reacts strongly with a mucus-like substance that is present around individual spheres in the aggregate IVS. Potential roles for the IVS in mediating X. nematophila colonization of the nematode intestine are discussed. PMID- 16309460 TI - The pilus and porin of Neisseria gonorrhoeae cooperatively induce Ca(2+) transients in infected epithelial cells. AB - Purified pili and porin from Neisseria quickly mobilize calcium (Ca(2+)) stores in monocytes and epithelial cells, ultimately influencing host cell viability as well as bacterial intracellular survival. Here, we examined the Ca(2+) transients induced in human epithelial cells during infection by live, piliated N. gonorrhoeae. Porin induced an influx of Ca(2+) from the extracellular medium less than 60 s post infection. The porin-induced transient is followed by a pilus induced release of Ca(2+) from intracellular stores. The timing of these events is similar to that observed using purified proteins. Interestingly, the porin induced Ca(2+) flux is required for the pilus-induced transient, indicating that the pilus-induced Ca(2+) release is, itself, Ca(2+) dependent. Several lines of evidence indicate that porin is present on pili. Moreover, pilus retraction strongly influences the porin- and pilus-induced Ca(2+) fluxes. These and other results strongly suggest that the pilus and porin cooperate to modulate calcium signalling in epithelial cells, and propose a model to explain how N. gonorrhoeae triggers Ca(2+) transients in the initial stages of pilus-mediated attachment. PMID- 16309461 TI - Enteropathogenic Escherichia coli (EPEC) effector-mediated suppression of antimicrobial nitric oxide production in a small intestinal epithelial model system. AB - In vivo studies with the mouse-specific member of the attaching and effacing (A/E) family of pathogens raised the possibility that these non-invasive enteric pathogens can specifically inhibit inducible nitric oxide synthase (iNOS) expression to prevent the production of antimicrobial nitric oxide (NO). In this study we use polarized Caco-2 cells, a model of human small intestinal epithelia, to (i) demonstrate conclusively that an A/E member, human specific enteropathogenic Escherichia coli (EPEC), can inhibit cytokine-induced iNOS expression, (ii) show that this activity is dependent on the delivery of effector molecules into host cells and (iii) investigate the mechanism of inhibition. Analysis of the level of iNOS-related mRNA, protein and NO production demonstrated that EPEC can inhibit iNOS expression at the transcriptional, by direct and indirect mechanisms, and post-transcriptional levels. This transcriptional block was linked to the failure of the iNOS-related transcriptional factor NF-kappaB, but not STAT1, to undergo phosphorylation associated activation. A selective pressure to prevent iNOS production was evidenced by the finding that iNOS activity had a potent antimicrobial effect on adherent but not non-adherent bacteria. Moreover, given the central role NF kappaB plays in transcribing genes associated with early host immune responses, this inhibitory mechanism presumably represents an important role in pathogenesis. Our study also provides insights into the nature of NO production in response to bacterial infection as well as the role of the locus of enterocyte effacement (LEE)-encoded effector molecules in inhibiting iNOS expression. PMID- 16309462 TI - Surface ultrastructure of SARS coronavirus revealed by atomic force microscopy. AB - Atomic force microscopy has been used to probe the surface nanostructures of severe acute respiratory syndrome coronavirus (SARS-CoV). Single crown-like virion was directly visualized and quantitative measurements of the dimensions for the structural proteins were provided. A corona of large, distinctive spikes in the envelope was measured after treatment with hydroxyoctanoic acid. High resolution images revealed that the surface of each single SARS-CoV was surrounded with at least 15 spherical spikes having a diameter of 7.29 +/- 0.73 nm, which is in close agreement with that of S glycoproteins earlier predicted through the genomes of SARS-CoV. This study represents the first direct characterization of the surface ultrastructures of SARS-CoV particles at the nanometre scale and offers new prospects for mapping viral surface properties. PMID- 16309463 TI - A three-dimensional tissue culture model for the study of attach and efface lesion formation by enteropathogenic and enterohaemorrhagic Escherichia coli. AB - We sought to develop a practical and representative model to study the interactions of enteropathogenic and enterohaemorrhagic Escherichia coli (EPEC and EHEC, respectively) with human intestinal tissue. For this purpose, human intestinal epithelial HCT-8 cells were cultured under low-shear microgravity conditions in a rotating cell culture system. After 10 days, layered cell aggregates, or 'organoids', developed. Three lines of evidence indicated that these organoids exhibited traits characteristic of normal tissue. First, the organoids expressed normal intestinal tissue markers in patterns that suggested greater cellular differentiation in the organoids than conventionally grown monolayers. Second, the organoids produced higher levels of intestinally expressed disaccharidases and alkaline phosphatase on a cell basis than did conventionally cultured monolayers. Third, HCT-8 organoid tissue developed microvilli and desmosomes characteristic of normal tissue, as revealed by electron microscopy. Because the low-shear microgravity condition is proposed by modelling studies to more closely approximate conditions in the intestinal microvilli, we also tested the impact of microgravity of bacterial growth and virulence gene expression. No influence on growth rates was observed but intimin expression by EHEC was elevated during culture in microgravity as compared with normal gravity. That the responses of HCT-8 organoids to infection with wild-type EPEC or EHEC under microgravitational conditions approximated infection of normal tissue was demonstrated by the classical appearance of the resultant attaching and effacing lesions. We concluded that the low shear microgravity environment promoted growth of intestinal cell organoids with greater differentiation than was seen in HCT-8 cells maintained in conventional tissue culture and provided a reduced gravity environment for study of bacterial-host cell interactions. PMID- 16309464 TI - Novel effects of the prototype translocating Escherichia coli, strain C25 on intestinal epithelial structure and barrier function. AB - Intestinal bacteria play an etiologic role in triggering and perpetuating chronic inflammatory bowel disorders. However, the precise mechanisms whereby the gut microflora influences intestinal cell function remain undefined. Therefore, the effects of the non-pathogenic prototype translocating Escherichia coli, strain C25 on the barrier properties of human T84 and Madine-Darby canine kidney type 1 epithelial cells were examined. T-84 cells were also infected with commensal E. coil, strains F18 and HB101, and enterohaemorrhagic E. coli, serotype O157:H7. Strains F18 and HB101 had no effect on transepithelial electrical resistance (TER) of T84 monolayers. By contrast, epithelial cells infected with strain C25 displayed a time-dependent decrease in TER, preceded by an altered distribution of the cytoskeletal protein alpha-actinin, comparable to infection with E. coli O157:H7. E. coli C25 infection also led to activation of nuclear factor kappaB (NF-kappaB), interleukin-8 secretion and alterations in localization of claudin 1, but not zona occludens-1 or claudin-4, in T84 cells. There were adherent C25 bacteria on the intact apical surface of infected T84 cells, while mitochondria appeared swollen and vacuolated. These novel findings demonstrate the ability of a translocating commensal bacterium to adhere to and modulate intestinal epithelial barrier function and to induce morphological changes in a manner distinct from the known enteric pathogen, E. coli O157:H7. PMID- 16309465 TI - Human mucosa/submucosa interactions during intestinal inflammation: involvement of the enteric nervous system in interleukin-8 secretion. AB - Interleukin-8 (IL-8) is a key chemokine upregulated in various forms of intestinal inflammation, especially those induced by bacteria such as Clostridium difficile (C. difficile). Although interactions between different mucosal and submucosal cellular components have been reported, whether such interactions are involved in the regulation of IL-8 secretion during C. difficile infection is unknown. Moreover, whether the enteric nervous system, a major component of the submucosa, is involved in IL-8 secretion during an inflammatory challenge remains to be determined. In order to investigate mucosa/submucosa interactions that regulate IL-8 secretion, we co-cultured human intestinal mucosa and submucosa. In control condition, IL-8 secretion in co-culture was lower than the sum of the IL 8 secretion of both tissue layers cultured alone. Contrastingly, IL-8 secretion increased in co-culture after mucosal challenge with toxin B of C. difficile through an IL-1 beta-dependent pathway. Moreover, we observed that toxin B of C. difficile increased IL-8 immunoreactivity in submucosal enteric neurones in co culture and in intact preparations of mucosa/submucosa, through an IL-1 beta dependent pathway. IL-1 beta also increased IL-8 secretion and IL-8 mRNA expression in human neuronal cell lines (NT2-N and SH-SY5Y), through p38 and ERK1/2 MAP kinase-dependent pathways. Our results demonstrate that mucosa/submucosa interactions regulate IL-8 secretion during inflammatory processes in human through IL-1 beta-dependent pathways. Finally we observed that human submucosal neurones synthesize IL-8, whose production in neurones is induced by IL-1 beta via MAPK-dependent pathways. PMID- 16309466 TI - Eicosanoid-mediated proinflammatory activity of Pseudomonas aeruginosa ExoU. AB - As Pseudomonas aeruginosa ExoU possesses two functional blocks of homology to calcium-independent (iPLA(2)) and cytosolic phospholipase A(2) (cPLA(2)), we addressed the question whether it would exhibit a proinflammatory activity by enhancing the synthesis of eicosanoids by host organisms. Endothelial cells from the HMEC-1 line infected with the ExoU-producing PA103 strain exhibited a potent release of arachidonic acid (AA) that could be significantly inhibited by methyl arachidonyl fluorophosphonate (MAFP), a specific PLA(2) inhibitor, as well as significant amounts of the cyclooxygenase (COX)-derived prostaglandins PGE(2) and PGI(2). Cells infected with an isogenic mutant defective in ExoU synthesis did not differ from non-infected cells in the AA release and produced prostanoids in significantly lower concentrations. Infection by PA103 induced a marked inflammatory response in two different in vivo experimental models. Inoculation of the parental bacteria into mice footpads led to an early increase in the infected limb volume that could be significantly reduced by inhibitors of both COX and lipoxygenase (ibuprofen and NDGA respectively). In an experimental respiratory infection model, bronchoalveolar lavage (BAL) from mice instilled with 10(4) cfu of PA103 exhibited a marked influx of inflammatory cells and PGE(2) release that could be significantly reduced by indomethacin, a non selective COX inhibitor. Our results suggest that ExoU may contribute to P. aeruginosa pathogenesis by inducing an eicosanoid-mediated inflammatory response of host organisms. PMID- 16309467 TI - The rhoptry neck protein RON4 re-localizes at the moving junction during Toxoplasma gondii invasion. AB - Host cell invasion in the Apicomplexa is unique in its dependency on a parasite actin-driven machinery and in the exclusion of most host cell membrane proteins during parasitophorous vacuole (PV) formation. This exclusion occurs at a junction between host cell and parasite plasma membranes that has been called the moving junction, a circumferential zone which forms at the apical tip of the parasite, moves backward and eventually pinches the PV from the host cell membrane. Despite having been described by electron microscopic studies 30 years ago, the molecular nature of this singular structure is still enigmatic. We have obtained a monoclonal antibody that recognizes the moving junction of invading tachyzoites of Toxoplasma gondii, in a pattern clearly distinct from those described so far for microneme and rhoptry proteins. The protein recognized by this antibody has been affinity purified. Mass spectrometry analysis showed that it is a rhoptry neck protein (RON4), a hypothetical protein with homologues restricted to Apicomplexa. Our findings reveals for the first time the participation of rhoptry neck proteins in moving junction formation and strongly suggest the conservation of this structure at the molecular level among Apicomplexa. PMID- 16309468 TI - Mustard: a potential agent of chemical warfare and terrorism. AB - As one of the most important vesicant agents, the destructive properties of mustards on the skin, eyes and respiratory system, combined with a lack of antidote, makes them effective weapons. Such weapons are inexpensive, easily obtainable and frequently stockpiled. Sulphur mustard (mustard gas) has been used as a chemical warfare agent in at least 10 conflicts. In this article, the use of mustard as a potential agent of chemical warfare and terrorism is outlined. The dose-dependent effects of acute sulphur mustard exposure on the skin, eyes, and respiratory system are described, as well as the possible extents of injuries, the mechanisms of action and the long-term complications. Prevention and management of mustard exposure are briefly discussed. The need for awareness and preparedness in the dermatological community regarding mustard exposure is emphasized. PMID- 16309469 TI - Infrared-monitored cold response in the assessment of Raynaud's phenomenon. AB - BACKGROUND: Evaluation of treatments for Raynaud's phenomenon (RP) requires objective response parameters in addition to clinical activity scores. Thermographic monitoring of fingertip re-warming after cold challenge has been widely used but usually requires sophisticated equipment. We have previously shown that fingertip re-warming after cold challenge follows a first-order transient response curve that can be described by a single variable, designated tau. OBJECTIVES: Here, we describe a novel device termed a duosensor, which records the tau value upon cold challenge in an automated manner. METHODS: We determined tau values in healthy probands, patients with primary or secondary RP associated with autoimmune disease and patients with scleroderma-associated RP following cold challenge, to determine assay variability, sensitivity and specificity. RESULTS: Duosensor-based thermography exhibited low intraindividual variability in healthy probands. As expected, tau values in RP patients were significantly increased compared with controls (8.08 +/- 3.65 min vs. 3.23 +/- 1.65 min). The duosensor-determined tau value yielded a specificity of 94.6% and predictive value of 95.3% for the presence of RP in a retrospective analysis of 139 patients. Furthermore, in a cohort of scleroderma patients with RP, patient self-assessment of RP severity correlated with tau values. CONCLUSIONS: Taken together, the present data suggest that tau value determination provides a suitable outcome measure for clinical studies of novel RP treatments. As the duosensor is a simple stand-alone device requiring no supporting equipment and minimal personnel attention, it should allow RP activity monitoring even in clinical settings with minimal technical infrastructure. PMID- 16309470 TI - Teledermatoscopy as a triage system for pigmented lesions: a pilot study. AB - BACKGROUND: Skin-cancer screening should rely on simple, low-cost and high sensitivity diagnostic procedures. AIMS: To compare diagnosis and management options after the evaluation of clinical and dermatoscopic teleconsultations with a store-and-forward teledermatology screening system for pigmented lesions. METHODS: Kappa values between telediagnoses and the gold standard (histological examination) were assessed. Referral rates, diagnostic confidence level (DCL), sensitivity and specificity were evaluated in both approaches. Picture quality and time and cost investments were also measured. In total, 61 teleconsultations were evaluated. RESULTS: Sensitivity of the clinical and dermatoscopic teleconsultations was 1 for both, whereas specificities were 0.65 and 0.78, respectively (P < 0.05). DCL was higher for the dermatoscopic teleconsultations (4.75 vs. 4.14, P < 0.05). Agreement between the clinical and dermatoscopic teleconsultation was kappa = 0.89 (95% CI 0.81-0.97). Agreement with the gold standard was 0.91 (95% CI 0.82-1.00) for the clinical teleconsultation and 0.94 (95% CI 0.88-1.00) for teledermatoscopy (P > 0.05). Teledermatoscopy increased the economic investment of a teledermatology facility by 2.4 times. The GP spent 1.5 times longer on dermatoscopic teleconsultations. CONCLUSIONS: Teledermatoscopy has improved the DCL, specificity and referral rates of a teledermatology-based screening system for pigmented lesions. A more detailed economic analysis remains to be performed before recommending teledermatoscopy as a routine screening procedure in pigmented-lesion clinics. PMID- 16309471 TI - Using the Dermatology Index of Disease Severity (DIDS) to assess the responsiveness of dermatitis. AB - BACKGROUND: The Dermatology Index of Disease Severity (DIDS) is designed as an instrument for staging severity of inflammatory dermatoses. It measures the therapeutic effectiveness and magnitude of clinical improvement in several types of dermatitis, eczema and psoriasis. When used in the Psoriasis Outcomes Study, it was simpler and easier than the Psoriasis Area and Severity Index. The reliability and validity of DIDS have already been established in our previous study. AIM: To test the responsiveness of the DIDS in patients with various types of dermatitis following treatment. METHODS: The DIDS was used to stage patients with dermatitis over time. In total, 142 patients with dermatitis were enrolled into the study. Only 55 patients completed the follow-ups on days 30 or 60, thus the data from these 55 patients only were analysed. The Wilcoxon signed rank test for paired comparisons was used to test for significance of shift in the individual's DIDS staging over the time period. RESULTS: 35 patients were seen on or close to day 30 after the initial visit, of whom 34 patients (76%) had an improvement of at least one stage on DIDS. There were 51 patients seen at or close to day 60, of whom 40 patients (78%) had an improvement of at least one stage on DIDS. The remaining 22-24% of patients did not improve, but neither did they worsen. The joint shift of each individual's DIDS staging from baseline to follow-up was statistically significant (P < 10(-5), two-sided t-test). CONCLUSION: The results of this study support the responsiveness of the DIDS for tracking severity of disease over time in patients with various types of dermatitis. PMID- 16309472 TI - Dandruff-associated smouldering alopecia: a chronobiological assessment over 5 years. AB - BACKGROUND: In some individuals, dandruff may be recurrent and even chronic. This represents a difficult-to-treat condition. Excessive hair shedding and diffuse alopecia may develop. OBJECTIVES: To evaluate the chronobiological aspects of dandruff, hair shedding and alopecia. METHODS: This study was performed in eight men suffering from such a scalp condition. They were examined 40 times at regular intervals for five consecutive years. Hair-density assessments and trichograms were performed at least twice per trimester on the parietal region of the scalp, corresponding to an area where dandruff was present. RESULTS: A positive correlation was found between the percentage of telogen hairs and dandruff severity as assessed by the squamometry index. Bimodal yearly biorhythms were found for dandruff severity, telogen counts and hair loss between two consecutive visits. The chronobiological fluctuations were prominent in some study participants, but remained inconspicuous in others. The biorhythms were almost synchronized in all participants, showing variations over the year. CONCLUSIONS: The biorhythms on the hair cycle in dandruff are not fundamentally different from those previously reported in subjects without dandruff. The periodicity may be an intrinsic feature of human skin. PMID- 16309473 TI - Laser remodelling of nodular nasal lupus pernio. AB - Disfiguring nodular nasal lupus pernio is a rare condition that responds poorly to medical management. We have treated three affected patients with carbon dioxide laser remodelling, and report their progress 6 years, 2 years and 16 months following treatment, respectively. Although the abnormal granulomatous tissue was debulked rather than completely excised, the wounds healed within 4 weeks in all patients. The cosmetic results are acceptable and have been maintained in two of the patients, perhaps a reflection of their overall systemic sarcoidosis control. PMID- 16309474 TI - Acquired port-wine stain related to acoustic neuroma. AB - Port-wine stains are frequently seen congenital vascular malformations consisting of ectatic dermal capillaries. Acquired port-wine stain that develops later in life is an uncommon vascular lesion that is morphologically identical to a congenital port-wine stain. In the majority of acquired port-wine stains, the aetiology is unknown, but trauma is an important causative factor. Other proposed aetiologies include chronic sun exposure, hormonal changes, frostbite injury, obstruction of the peritoneovenous shunt, herpes zoster infection, and cerebral arteriovenous malformation. Here we report the first case of a patient who had an acquired port-wine stain related to a solid brain tumour. PMID- 16309475 TI - A case of drug-induced hypersensitivity syndrome showing transient immunosuppression before viral reactivation during treatment for pemphigus foliaceus. AB - Drug-induced hypersensitivity syndrome (DIHS) is one of the most severe drug adverse reactions, with characteristic biphasic symptoms. Reactivation of human herpesvirus-6 (HHV-6) is frequently observed, although the cause of DIHS is still unknown. A patient developed DIHS during treatment with diaminodiphenylsulphone for pemphigus foliaceus. The number of lymphocytes in his peripheral blood, and titres of serum total IgG and IgM and anti-desmoglein1 antibody transiently decreased just before reactivation of HHV-6, cytomegalovirus and Epstein-Barr virus. This observation suggests that transient suppression of both cellular and humoral immunity may trigger viral reactivation, which leads to the development of the second phase of DIHS. PMID- 16309476 TI - Photosensitive pityriasis rubra pilaris. AB - Exacerbation of pityriasis rubra pilaris (PRP) with ultraviolet exposure is well recognized. However, the initial presentation of PRP in a photoexposed distribution is infrequently reported, and only rarely confirmed with phototesting. We describe such a case of photosensitive PRP. Phototesting revealed exquisite ultraviolet (UV) sensitivity, which was particularly marked to UVB, a feature shared with previously reported cases. PMID- 16309477 TI - Iatrogenic phytophotodermatitis resulting from herbal treatment of an allergic contact dermatitis. AB - Phytophotodermatitis commonly occurs in skin exposed to sunlight after contact with plants containing furocoumarins. While it is recognized that the consumption of plants or vegetables containing furocoumarins can potentially trigger a phytophotodermatitis, there have been no reports to date of a phytophotodermatitis triggered by an ingested herbal remedy. We describe the case of a 56-year-old farmer who developed an extensive photo-induced dermatitis after ingesting a herbal decoction prescribed for his chronic hand dermatitis. PMID- 16309478 TI - A reticular variant of elastolytic giant cell granuloma. AB - An 80-year-old man presented with a distinctive reticular form of elastolytic giant cell granuloma (EGCG). This represents a rare subset of EGCG, thus belonging to a larger group of granulomatous skin diseases with poorly understood pathogenesis. The patient exhibited numerous erythematous to brownish, sharply demarcated patches of reticulate appearance with partial central atrophy and nonelevated margins, which involved the trunk and especially chest, shoulders, upper back and arms. Skin biopsy revealed a granulomatous infiltrate with multinucleate histiocytes containing remnants of elastic fibres in conjunction with a significant loss of elastic fibres throughout the dermis. There is little published information on treatment, which remains unsatisfactory. PMID- 16309479 TI - Retrospective diagnosis of Kindler syndrome in a 37-year-old man. AB - Kindler syndrome is a rare autosomal recessive disorder characterized by acral blisters in infancy and early childhood, followed by photosensitivity, progressive poikiloderma and cutaneous atrophy. Other features include webbing of the toes and fingers, palmoplantar hyperkeratosis, gingival fragility, poor dentition, and mucosal involvement in the form of urethral, anal and oesophageal stenosis. The recent finding of KIND1 mutations in Kindler syndrome facilitates early diagnosis, prophylactic measures and more precise definition of the phenotype. In the family described here, molecular diagnosis of Kindler syndrome in an infant with acral blisters led to the belated diagnosis in a severely affected relative whose condition had remained unidentified for 37 years. PMID- 16309480 TI - Keratosis lichenoides chronica: a diagnostic and therapeutic challenge. AB - A 45-year-old man presented with a 7-year history of keratosis lichenoides chronica (KLC), a mucocutaneous lichenoid disorder of keratinization with no known aetiology, without significant systemic associations. Our patient also had type I diabetes mellitus, mild hypertension and lipid abnormalities. The diagnosis and treatment of KLC is often challenging. PMID- 16309481 TI - Photodynamic therapy of genital condylomata in men. AB - Current treatments for genital condylomata are not completely satisfactory, as they fail to clear lesions in a proportion of patients, and relapses after successful treatment are frequently seen. Photodynamic therapy (PDT) using topical 5-aminolaevulinic acid (5-ALA) has been suggested as a novel treatment option. We performed a small open study using topical 5-ALA and red light (630 nm) in nine men with genital condylomata and a history of at least one previous unsuccessful conventional treatment. Complete cure was achieved in three patients, one of whom experienced a relapse after 3 weeks. Three patients showed partial responses, and three showed no response. Based on the currently available evidence, PDT is a viable treatment option for selected cases that fail to respond to other therapies. PMID- 16309482 TI - Tyrosinaemia type II with diffuse plantar keratoderma and self-mutilation. AB - Tyrosinaemia type II (oculocutaneous tyrosinaemia or Richner-Hanhart syndrome) is a rare and potentially treatable genetic disorder. Corneal opacities or ulcers, circumscribed palmoplantar keratoderma and subnormal intelligence form the classical triad of this syndrome. A case with additional features of self-harm and diffuse plantar keratoderma is discussed. Emphasis is laid on the importance of early diagnosis and intervention in the form of dietary regulation to prevent the neurological signs and treat the oculocutaneous symptoms. PMID- 16309483 TI - Bacterial infection-induced generalized Hailey-Hailey disease successfully treated by etretinate. AB - Hailey-Hailey disease (HHD; familial benign chronic pemphigus) is a hereditary blistering disorder characterized by episodic maceration and erosions mainly in intertriginous areas, and generalized eruptions are rarely seen. We report here a 51-year-old woman with generalized HHD who was successfully treated with oral etretinate. The present case suggests that oral etretinate is effective against the generalized eruptions even in cases in which bacterial infection has triggered the generalization of HHD. PMID- 16309484 TI - Microscopic polyarteritis presenting with skin necrosis in a patient with sickle cell disease. AB - A 20-year-old Caribbean woman with sickle cell anaemia was admitted with a 4-day history of fever and a painful swollen right ankle. She rapidly developed skin necrosis. The differential diagnosis is discussed. This case illustrates the difficulty in identifying the cause of cutaneous necrosis in an acutely ill patient. In our patient, histopathology implicated a vasculitic process, which was subsequently identified as a manifestation of microscopic polyarteritis. PMID- 16309485 TI - Childhood cicatricial pemphigoid confined to the vulva. AB - Cicatricial pemphigoid or benign mucous membrane pemphigoid is an autoimmune bullous disease predominantly affecting the mucosal surfaces and healing with scar formation. Localized cicatricial pemphigoid of the vulva in children is rare. We present a child with this rare condition who was initially investigated on suspicion of her being subjected to child sexual abuse, and discuss its management. PMID- 16309486 TI - Long-term results of topical PUVA in necrobiosis lipoidica. AB - The aim of our study was to evaluate the long-term results of topical psoralen ultraviolet A (PUVA) in patients with necrobiosis lipoidica (NL), in whom conventional methods (pentoxifylline, vitamin E, tretinoin, and topical or intralesional corticosteroids) had failed. The study comprised 10 women (age range 17-44 years), six of whom were insulin-dependent diabetics and four were diabetes-free. Duration of NL ranged from 3 to 10 years. The patients were treated with a 0.005% aqueous solution of 8-methoxypsoralen, applied topically for 30 min, and subsequently irradiated with UVA three times weekly. All the patients experienced almost complete remission (softening of skin lesions, no hyperpigmentation, lack of lesion progression) after a mean of 47 sessions (mean UVA cumulative dose 69.5 J/cm2). They were followed up for 12-24 months, during which time two recurrences, both in diabetic patients, were observed after 8 and 12 months of treatment cessation, which further resolved after another course of topical PUVA. We conclude that topical PUVA is well tolerated by NL patients and may serve as an alternative therapeutic regimen. PMID- 16309487 TI - Two cases of folliculosebaceous cystic hamartoma. AB - Folliculosebaceous cystic hamartoma (FSCH) is a rare cutaneous hamartoma composed of dilated folliculosebaceous units associated with mesenchymal elements. Two cases of FSCH with typical histopathological features are reported. Patient 1 was a 60-year-old man presented with a normal skin-coloured asymptomatic nodule on his scalp. Patient 2 was a 70-year-old man with an asymptomatic nodule on his right auricle that had persisted for the previous 15 years. In all, 34 cases of FSCH have been reported in the English literature. Clinically, the lesions are asymptomatic, usually rubbery to firm in consistency, and usually occur on or above the neck (> 90%). Most lesions do not exceed 25 mm in diameter (> 90%). Histopathologically, FSCH shares several similar features to sebaceous trichofolliculoma, but it is usually possible to differentiate these two tumours. PMID- 16309488 TI - Epidermolysis bullosa acquisita: diagnosis by fluorescence overlay antigen mapping and clinical response to high-dose intravenous immunoglobulin. AB - Epidermolysis bullosa acquisita (EBA) is a chronic, subepidermal blistering disease characterized by the presence of autoantibodies to type VII collagen, located below the lamina densa of the basement membrane zone (BMZ). There is a large clinical and histological overlap between EBA and other subepidermal autoimmune bullous diseases, therefore, complex immunological techniques are required to make an accurate diagnosis. Therapy of EBA is also a difficult issue. Most patients do not respond to several common immunosuppressive agents. We describe a patient who has shown a good response to high-dose intravenous immunoglobulin therapy. PMID- 16309489 TI - Multiple nodules on the face and in the nasal cavity are the symptoms of vegetative pyoderma gangrenosum complicated with myelodysplastic syndrome. AB - We have encountered and treated an unusual case of pyoderma gangrenosum (PG) characterized by multiple nodular lesions on the face and in the nasal meatus, which was complicated by myelodysplastic syndrome. PMID- 16309490 TI - Discoid lupus erythematosus of the eyelids associated with staphylococcal blepharitis and Meibomian gland dysfunction. AB - Lower eyelid involvement occurs in 6% of patients with discoid lupus erythematosus (DLE). Eyelid lesions are rarely the initial manifestation of DLE. We describe a 25-year-old woman presenting with discoid lesions of the lower eyelids, staphylococcal blepharitis and Meibomian gland dysfunction, who later developed a discoid lesion on the chin. Histopathological and immunofluorescence studies of a biopsy specimen from this lesion established the diagnosis of DLE. We are unaware of any previously reported cases of DLE presenting with discoid eyelid lesions associated with staphylococcal blepharitis and Meibomian gland dysfunction. DLE should be considered as a differential diagnosis in chronic blepharitis that persists despite usual medical management and eyelid hygiene. Misdiagnosis may lead to eyelid margin deformities, necessitate a complicated full-thickness biopsy, and delay diagnosis of systemic lupus. PMID- 16309491 TI - Vitiligo with inflammatory raised borders, associated with atopic dermatitis. AB - A 31-year-old man had had atopic dermatitis since childhood and developed vitiligo with inflammatory raised borders 5 years prior to presentation. Immunohistochemically, CD4+ T cells infiltrated predominantly in the raised border of vitiligo, while CD8+ T cells were present just outside of the borders, suggesting that CD8+ cells were an antecedent to the CD4+ cells. Despite the presence of atopic dermatitis, the percentage of CXCR3+ CD4+ Th1 cells increased in the patient's peripheral blood, compared with a representative atopic patient showing a high percentage of CCR4+CD4+ Th2 cells. This case suggests that vitiligo with inflammatory raised borders can occur even in patients with atopic dermatitis when Th1 cells are activated and overcome the Th2-dominant state. PMID- 16309492 TI - Aggressive NK-cell leukaemia associated with reactive haemophagocytic syndrome. AB - We report a case of aggressive NK-cell leukaemia associated with reactive haemophagocytic syndrome in a 29-year-old Korean woman who had several small purpuric patches on both thighs. She also had high fever. Laboratory tests revealed pancytopenia and deranged liver function, and atypical lymphocytes containing toxic granules were detected from peripheral blood and bone marrow. The bone marrow examination showed diffuse histiocytic proliferation with several haemophagocytic macrophages, suggesting an associated reactive haemophagocytic syndrome. Skin biopsy from her thigh lesion demonstrated atypical CD56+ lymphoid cellular infiltrates with angiocentric pattern, and in situ hybridization test for Epstein-Barr virus was positive. Although we treated her with chemotherapy, she died 1 month later. PMID- 16309493 TI - Diffuse trichostasis spinulosa in chronic renal failure. AB - Trichostasis spinulosa is a relatively common but underdiagnosed disorder of the pilosebaceous follicles in which there is follicular hyperkeratosis of a dilated vellus hair follicle with retention of successive telogen hairs arranged in parallel among keratinaceous material. The condition is apparent as dark follicular plugs in the affected areas, most commonly affecting the face (nose and cheeks) or the trunk (interscapular area in particular). We describe a 33 year-old Indian man who was on haemodialysis for chronic renal failure and who developed unusually widespread trichostasis spinulosa over almost the entire body including the lower limbs over a period of 18 months. PMID- 16309494 TI - The pathophysiology of atopic eczema. AB - Atopic eczema (AE) represents a pruritic chronic inflammatory skin disease with a complex background, triggered by genetic and environmental factors. Different dendritic cells subtypes, such as Langerhans cells, inflammatory dendritic epidermal cells and plasmacytoid dendritic cells, play a key role in AE and impact on the mechanisms underlying AE, such as the recruitment of inflammatory cells, T-cell priming, and cytokine and chemokine release. In addition, allergens in combination with bacterial and viral stimuli influence the course and severity of AE. In this review, we highlight the recent progress made in the pathophysiology of AE focusing on the latest research results published in this field. PMID- 16309495 TI - Endogenous production of nitric oxide contributes to proliferation effect of vascular endothelial growth factor-induced malignant melanoma cell. AB - The objectives of this study were to observe the effect of overexpression of vascular endothelial growth factor (VEGF) on the proliferation of the malignant melanoma (MM) cell line A375, and to study the role of nitric oxide (NO) in this process and the mechanism of VEGF induced-A375 cell proliferation. The VEGF(165) cDNA was transfected into A375 cells by electroporation. VEGF mRNA and protein in A375 cells were detected by RT-PCR and ELISA. The proliferation of A375 cells was assessed by cell counting and MTT assay. Protein expression of iNOS, eNOS and nNOS was detected by Western blotting. NO production in A375 cell supernatant was measured by the nitrate reductase method. VEGF mRNA in A375 cells was significantly increased 72 h and 96 h after transfection of VEGF(165) cDNA, as were VEGF protein, NO and iNOS levels. However, protein expression of eNOS and nNOS was not detected in either transfected or untransfected cells. Proliferation of A375 cells transfected with VEGF(165) cDNA was enhanced. The nitric oxide synthase inhibitor l-NAME could dose-dependently inhibit the proliferation of A375 cells evoked by VEGF. These results indicate that VEGF enhances the expression of iNOS in A375 cells and results in an increase in NO formation, which may be important in the process of VEGF-induced proliferation of A375 cells. PMID- 16309496 TI - Overexpression of the suppressor of cytokine signalling 3 (SOCS3) in severe atopic dermatitis. AB - Immunoregulatory conditions in atopic dermatitis (AD) involve both Th1 and Th2, although primarily Th2. The suppressor of cytokine signalling 3 (SOCS3) may mediate Th2 regulatory response through negative regulation of Th1 pathways. We examined SOCS3 expression in skin lesions of patients with severe AD and compared it with that in patients with the Th1 disorder, psoriasis. We assessed SOCS3 expression in skin lesions from seven patients with severe AD and five patients with typical psoriasis, and in skin from three normal subjects, using immunohistochemical staining, in situ hybridization and reverse transcription PCR. Strong SOCS3 expression was found in skin lesions in all patients with AD, but only weak expression in patients with psoriasis and in normal subjects. Overexpression of SOCS3 is prominent in the skin of patients with severe AD and consequently may prove useful for evaluating the severity of AD as a Th2-dominant condition. PMID- 16309497 TI - High serum galectin-3 in advanced melanoma: preliminary results. AB - BACKGROUND: Galectin-3 (Gal-3) is a member of the family of beta-galactoside binding mammalian lectins, and has been implicated in tumour invasion and metastatic process in vitro and in vivo. AIM: To determine whether an increase in serum Gal-3 production could be found in patients with advanced metastatic melanoma. METHODS: We collected 18 sera from patients with AJCC stage IV metastatic melanomas and 20 sera from healthy volunteers. Determination of Gal-3 was performed by ELISA, and in the group of patients with melanoma, these results were compared with the serum lactate dehydrogenase (LDH) and the C-reactive protein (CRP) concentrations. RESULTS: Gal-3 concentration was shown to be significantly higher in the group of patients with melanoma compared with healthy volunteers, and Gal-3 concentration was significantly correlated with both LDH and CRP in the melanoma group. We also selected four patients in the melanoma group for Gal-3 retrospective immunostaining analysis on cutaneous metastases. Two of these patients, who had a higher Gal-3 serum level, showed more intense staining and the other two patients, with a lower serum level of Gal-3, had moderate immunostaining, suggesting that at least part of serum Gal-3 might be produced by metastatic melanoma tissue. CONCLUSIONS: Gal-3 might play a role in melanoma progression and/or inflammation, and warrants further study. PMID- 16309498 TI - Audiological abnormalities in patients with vitiligo. AB - BACKGROUND: Accumulating evidence suggests that vitiligo is a systemic disease affecting the entire pigmentary system. AIM: To investigate the subclinical abnormalities of melanin-containing cellular elements of the auditory system in patients with vitiligo. METHODS: We studied the conventional audiometric investigations and brainstem auditory evoked responses (BAERs) of 57 active patients with vitiligo and 50 healthy human subjects. The I, III and V latencies, and I-III, III-V and I-V interpeak latencies (IPL) between the groups were compared. RESULTS: A mild degree of sensorineural hypoacusis was found in eight patients with vitiligo (14%), whereas no controls demonstrated abnormal audiological results (P =0.006). A statistically significant increase in both ears of the third peak latency (P =0.02, P = 0.01, respectively) and IPL I-III (P = 0.04, P = 0.008, respectively), and a significant increase of the fifth peak latency in the right ear (P = 0.04) were found, compared with controls, but no differences were found for other latencies and IPLs. CONCLUSIONS: Melanin may play a significant role in the establishment and/or maintenance of the structure and function of the auditory system and may modulate the transduction of the auditory stimuli by the inner ear. PMID- 16309499 TI - Inducible nitric oxide synthase and cytokine pattern in lesions of patients with American cutaneous leishmaniasis. AB - American tegumentary leishmaniasis has three forms: localized (LCL), found in resistant individuals; diffuse (DCL), found in susceptible individuals; and intermediate cutaneous leishmaniasis (ICL), found in individuals with exacerbated immunity. We evaluated cytokines and inducible nitric oxide synthase (iNOS) in lesions of LCL, ICL and DCL using immunohistochemistry. LCL granulomas showed a preponderance of interferon (IFN)-gamma and interleukin (IL)-12 expression, whereas ICL granulomas had more IL-4-, IL-10- and mainly transforming growth factor (TGF)-beta1-expressing cells. Higher densities of iNOS+ cells were observed in ICL and LCL than in DCL. iNOS was also expressed in keratinocytes of LCL and ICL lesions, and in epidermal dendritic cells of ICL lesions. In LCL and ICL, most keratinocytes expressed IL-12 and a portion expressed IFN-gamma. IL-12+ and IFN-gamma+ dendritic cells were absent or sparse in LCL and ICL epidermis. Our results show the importance of iNOS, IL-12 and INF-gamma in LCL and ICL lesions, emphasizing the existence of a mixed cytokine pattern in ICL different from the Th1 and Th2 responses established in LCL and DCL lesions. PMID- 16309500 TI - Multiple cutaneous and uterine leiomyomata resulting from missense mutations in the fumarate hydratase gene. AB - Multiple cutaneous and uterine leiomyomata (MCL) is an autosomal dominant disorder characterized by the development of benign smooth muscle tumours (leiomyomas) in the skin and uterus of affected women, and in the skin of affected men. In rare cases, MCL has been associated with a predisposition to the rare type II papillary renal cell cancer, also known as hereditary leiomyomatosis and renal cell cancer. The genetic locus for MCL has been mapped to chromosome 1q42.3-43 and subsequently, germline mutations in the fumarate hydratase (FH) gene have been identified. In addition, analysis of FH in some tumours of MCL patients revealed a second mutation inactivating the wild-type allele, suggesting that FH may function as a tumour suppressor gene. Here, we report two cases of MCL patients with FH mutations, designated as T287P and R190L. T287P represents a novel mutation of a highly conserved amino acid of the FH protein. In addition, a patient with an unusual clinical presentation of MCL was found to have the recurrent mutation, R190L, raising the possibility of incorporating FH sequencing as a diagnostic tool. Our findings extend the allelic series of mutations in FH and support its status as the underlying cause of MCL. PMID- 16309501 TI - Antifungal susceptibility and genetic similarity of sequential isolates of Trichophyton rubrum from an immunocompetent patient with chronic dermatophytosis. AB - Chronic cutaneous dermatophytoses caused by Trichophyton rubrum are common in immunocompromised patients. In immunocompetent indivuals, the disease is more often associated with onychomycosis and tinea pedis. The aim of this study was to perform antifungal susceptibility tests and genetic analysis of sequential isolates of T. rubrum from an immunocompetent patient with chronic dermatophytosis. Antifungal susceptibility tests against griseofulvin, ketoconazole, itraconazole and fluconazole were performed with sequential isolates of T. rubrum. Genetic relationship among the isolates was analysed by the random amplification of polymorphic DNA (RAPD) method. The results revealed that treatment failure was not related to the development of drug resistance, as all of the sequential T. rubrum isolates were sensitive to antifungals tested in vitro. The RAPD data demonstrated that this disease was caused by identical isolates, with no genetic differences among them, representing a single T. rubrum strain. Treatment failure and chronicity of infection do not seem to be related to antifungal resistance. PMID- 16309502 TI - Transcripts of the human herpesvirus 8 genome in skin lesions and peripheral blood mononuclear cells of a patient with classic Kaposi's sarcoma. AB - We report a 75-year-old Japanese woman with classic Kaposi's sarcoma. PCR amplified human herpesvirus 8 (HHV-8) DNA sequences from her skin lesions and peripheral blood mononuclear cells (PBMC), but not her plasma, saliva or urine. An antibody test against HHV-8 lytic antigens was positive. Immunohistochemical staining detected latent antigen. There was no evidence of HHV-8 infection in her husband, sister or daughter. Genes coding for HHV-8-encoded viral interleukin-6, viral macrophage inflammatory protein I, viral G protein-coupled receptor, viral cyclin D and viral Bcl-2 were expressed to the same degree in both her skin lesion and PBMC. Latency-associated T0.7 mRNA and HHV-8-encoded viral tegument protein genes were expressed in her PBMC at levels lower than in the skin lesions. Based on the gene expression profile, we concluded that lytic HHV-8 infection was present in her skin lesions and PBMC. PMID- 16309504 TI - A short history of Clinical and Experimental Dermatology. PMID- 16309505 TI - Anetoderma secondary to cutaneous B-cell lymphoma. PMID- 16309506 TI - Eosinophilic fasciitis secondary to treatment with atorvastatin. PMID- 16309507 TI - General practitioners' opinions regarding the need for training in dermatology at undergraduate and postgraduate levels. PMID- 16309508 TI - Precipitation of generalized pustular psoriasis by prednisolone. PMID- 16309509 TI - A case of cutis verticis gyrata, induced by misuse of anabolic substances? PMID- 16309510 TI - Where has all that hair gone? PMID- 16309511 TI - Erythema multiforme associated with acute hepatitis B virus infection. PMID- 16309512 TI - Gynaecomastia with oligospermia: an unusual complication of low-dose methotrexate for pustular psoriasis. PMID- 16309513 TI - Generalized exacerbation of psoriasis associated with imiquimod cream treatment of superficial basal cell carcinomas. PMID- 16309514 TI - Chronic graft-versus-host disease with skin signs suggestive of dermatomyositis. PMID- 16309515 TI - Contact dermatitis in guitar players. PMID- 16309516 TI - Response of pemphigus vulgaris to anti-CD20 antibody therapy (rituximab) may be delayed. PMID- 16309517 TI - Recalcitrant hand pompholyx: variable response to etanercept. PMID- 16309519 TI - Traumatic nail tale. PMID- 16309518 TI - Treatment of basal cell carcinoma with topical methylaminolaevulinate photodynamic therapy in an organ-transplant recipient. PMID- 16309520 TI - Propolis, beeswax, and the sensitization potential of topical calcineurin inhibitors. PMID- 16309521 TI - Fatal toxic epidermal necrolysis due to lansoprazole. PMID- 16309522 TI - Congenital smooth muscle hamartoma presenting with an unusual pseudo-Darier's sign. PMID- 16309523 TI - Kissing cafe-au-lait macule of the eyelids: report of the first case. PMID- 16309524 TI - A patient with subaural arteriovenous malformation complicated by macrotia. PMID- 16309525 TI - Pyoderma gangrenosum treated with anti-TNF alpha therapy (etanercept). PMID- 16309526 TI - Splinter haemorrhages associated with oral terbinafine in a Chinese man. PMID- 16309527 TI - Treatment of recalcitrant hidradenitis suppurativa with oral ciclosporin. PMID- 16309528 TI - A sandfly in Surrey? A case of cutaneous leishmaniasis in the United Kingdom without history of recent travel to an endemic area. PMID- 16309529 TI - Skin-coloured lumps in childhood. PMID- 16309530 TI - Multiple subcutaneous nodules with an infrapatellar mass. PMID- 16309531 TI - A swollen lip. PMID- 16309532 TI - Blistering plaques. PMID- 16309533 TI - Tender papules around a stoma. PMID- 16309534 TI - A cyst with inner meaning. PMID- 16309535 TI - Recent advances in haematology and oncology: graft-versus-host disease, classification and treatment of non-Hodgkin's lymphomas, and sentinel-node biopsy in breast cancer. PMID- 16309536 TI - Basal cell carcinoma lurking within gross rhinophyma. PMID- 16309537 TI - Second cite. PMID- 16309540 TI - British Society for Matrix Biology Spring 2005 meeting. University of Liverpool. Collagen--from genes to fibrils. PMID- 16309542 TI - A novel 3-dimensional culture system as an in vitro model for studying oral cancer cell invasion. AB - Tissue microenvironment plays a critical role in tumour growth and invasion. This study established a novel 3-dimensional (3-D) cell invasion model for direct microscopic observation of oral cancer cell invasion into the underlying basement membrane and connective tissue stroma. A multilayer cell construct was developed using the OptiCell chamber, consisting of a lower layer of oral mucosa fibroblasts embedded in collagen gel and an overlaying upper layer of oral cancer cells. The two layers are separated by a basement membrane composed of reconstituted extracellular matrix. To verify the applicability of the cell invasion model, multilayer cell constructs of oral squamous cell carcinoma and oral mucosal fibroblasts were exposed to extrinsic urokinase-type plasminogen activator (uPA) or plasminogen activator inhibitor (PAI-1), which are known effectors of cell migration. In addition, the constructs were exposed to both normoxic and hypoxic culture conditions. Microscopic study showed that the presence of uPA enhanced cell invasion, while PAI-1 inhibited cell migration. Western blot and zymographic analysis demonstrated that hypoxia up-regulated uPA and matrix metalloproteinases (MMPs) expression and activity; conversely, PAI-1 level was down-regulated in response to hypoxic challenge as compared to normoxic condition. Our results indicated that the novel 3-D invasion model could serve as an excellent in vitro model to study cancer cell invasion and to test conditions or mediators of cellular migration. PMID- 16309541 TI - Molecular predictors of clinical outcome in patients with head and neck squamous cell carcinoma. AB - Head and neck squamous cell carcinoma (HNSCC) involves the upper aerodigestive tract and can destroy the structure and function of organs involved in voice, speech, taste, smell and hearing, as well as vital structures necessary for survival. HNSCC has long been a treatment challenge because of the high rate of recurrences and of advanced disease at the time of diagnosis. Molecular identification of tissue biomarkers in diagnostic biopsy specimens may not only identify patients at risk for developing HNSCC but may also select patients that may benefit from more aggressive treatment modalities. Several biomarkers studied to date such as the proteins p53, cyclin D1, p16, Cox-2 enzyme, epidermal growth factor and vascular endothelial growth factor receptors, matrix metalloproteinases and the Fhit marker for genomic instability could be manipulated for the therapeutic benefit of these patients. This review presents the most updated information on molecular biomarkers with the greatest prognostic potential in HNSCC and discusses some factors that contribute to the controversy concerning their prognostic importance. PMID- 16309543 TI - Abnormal expression of bcl-2 and bax in rat tongue mucosa during the development of squamous cell carcinoma induced by 4-nitroquinoline 1-oxide. AB - 4-Nitroquinoline 1-oxide (4NQO)-induced rat tongue carcinogenesis is a useful model for studying oral squamous cell carcinoma. The aim of this study was to investigate the expression of bcl-2 and bax during tongue carcinogenesis induced by 4NQO. Male Wistar rats were distributed into three groups of 10 animals each and treated with 50 ppm 4NQO solution through their drinking water for 4, 12 or 20 weeks. Ten animals were used as negative control. Although no histological changes were induced in the epithelium after 4 weeks of carcinogen exposure, bcl 2 and bax were over-expressed (P < 0.01) in all layers of the 'normal' epithelium. The expression levels were the same in all layers of epithelium for both the antibodies used (bcl-2 or bax). In dysplastic lesions at 12 weeks following carcinogen administration, the levels of bcl-2 and bax expression did not increase when compared to negative control with the immunoreactivity for bcl 2 being restricted to the superficial layer of epithelium. In well-differentiated squamous cell carcinoma induced after 20 weeks of treatment with 4NQO, bcl-2 was expressed in some cells of tumour islands. On the other hand, immunostaining for bax was widely observed at the tumour nests. The labelling index for bcl-2 and bax showed an increase (P < 0.05) after only 4 weeks of 4NQO administration. In conclusion, our results suggest that abnormalities in the apoptosis pathways are associated with the development of persistent clones of mutated-epithelial cells in the oral mucosa. Bcl-2 and bax expression appears to be associated with a risk factor in the progression of oral cancer. PMID- 16309544 TI - The biochemical and morphological alterations following administration of melatonin, retinoic acid and Nigella sativa in mammary carcinoma: an animal model. AB - Worldwide, breast cancer is the second leading cause of cancer death among women and the third most common cancer. Although our understanding of the molecular basis of this fatal disease has improved, this malignancy remains elusive. Melatonin (Mel), retinoic acid (RA) and Nigella sativa (NS) are substances with anticancer effects. To date, our understanding of the mechanisms of therapeutic effects of these products in mammary cancer is still marginal. To look at the preventive and therapeutic values of these products, we carried out this investigation. An animal model formed of 80 rats was established. The animals were divided into eight groups of 10 animals each: (a) control group injected with the same vehicle used for treatments in the relevant dosages and routes; (b) carcinogen group injected with the known carcinogenic substance 7,12-di methylbenz(a)anthracene (DMBA) that induces mammary carcinoma; (c) three prophylactic (Pro) groups (Mel-Pro, RA-Pro and NS-Pro) injected with test substances (Mel, RA and NS, respectively) 14 days before the intake of the carcinogenic substance DMBA and then continued until the end of the experiments; and (d) three treated (Tr) groups (Mel-Tr, RA-Tr and NS-Tr) injected with the vehicles after the intake of DMBA. In both the Pro and Tr groups, the drugs were daily administered for 3 months. The animals were killed, and their serum and tissues were evaluated for (a) markers of tumorigenicity [serum levels of total sialic acid (TSA) and lipid-bound sialic acid (LSA)], (b) markers of endocrine derangement (serum prolactin, estradiol and progesterone levels), (c) apoptotic changes [serum tumour necrosis factor (TNF)-alpha, tissue caspase-3 activity, percentage of DNA fragmentation and ultrastructural features of apoptosis] and (d) markers of oxidative stress (tissue levels of lipid peroxides and nitric oxide). Carcinoma was absent both in the control and in the NS-Pro groups. Mammary carcinoma occurred in DMBA and other Pro and Tr groups. The frequency of mammary carcinoma was high in the carcinogen DMBA group (60%), followed by the Tr (56%) and finally the Pro groups (33%). These tumours included papillary, comedo and cribriform carcinomas. As compared with the control group, the development of carcinoma in the carcinogen DMBA group was associated with increased levels of (a) markers of tumorigenicity (77.0 +/- 3.3 vs. 209.0 +/- 5.6 and P < 0.05 for TSA; 28.7 +/- 1.7 vs. 41.8 +/- 1.2 and P < 0.01 for LSA), (b) markers of endocrine derangement (2.5 +/- 0.1 vs. 3.6 +/- 0.3 and P < 0.05 for prolactin; 39.6 +/- 1.3 vs. 24.8 +/- 2.1 and P < 0.01 for progesterone and 31.0 +/- 0.7 vs. 51.1 +/- 3.4 and P < 0.01 for estradiol) and (c) markers of oxidative stress (2.3 +/- 0.2 vs. 5.2 +/- 0.7 and P < 0.01 for lipid peroxides and 4.4 +/- 0.2 vs. 7.6 +/- 0.8 and P < 0.01 for nitric oxide). Also, it was associated with decreased levels of markers of apoptotic activity (20.8 +/- 1.1 vs. 13.4 +/- 0.7 and P < 0.01 for caspase-3; 29.0 +/- 1.7 vs. 20.9 +/- 1.3 and P < 0.05 for percentage of DNA fragmentation; and 9.4 +/- 0.8 vs. 52.1 +/- 3.3 and P < 0.01 for TNF-alpha). When compared with the carcinogen DMBA group, the development of carcinoma in the Pro and Tr groups was associated with decreased levels of (a) markers of tumorigenicity, (b) markers of endocrine derangement and (c) markers of oxidative stress. Alternatively, carcinogenicity was associated with statistically significant (P < 0.01) increased levels of markers of apoptotic activity. To conclude, the administration of Mel, RA and NS reduced the carcinogenic effects of DMBA, suggesting a protective role. The possible underlying mechanisms of these effects await further investigations. PMID- 16309545 TI - Neoplastic transformation and angiogenesis in the thymus of transgenic mice expressing SV40 T and t antigen under an L-pyruvate kinase promoter (SV12 mice). AB - Using several techniques, we have assessed morphological characteristics of a malignant thymic tumour in SV12 transgenic (Tg) mice expressing SV40 T and t antigens under control of an L-PK promoter. We describe the development of a carcinoma originating from thymic hyperplasia and followed by the formation of a benign tumour composed chiefly of medullary epithelial cells expressing the transgene and of lymphocytes, a pathology very rarely reported in mice. Our study of the SV12 Tg mice represents the first description of a model of a pure malignant thymic tumour associated with extensive angiogenesis maintained in numerous descendants. The formation of a large tumoral neovascular network, observed here, has never been described in human and/or experimental thymic tumours. Tumoral transformation and angiogenesis are demonstrated by immunolabelling with antibodies against various cytokeratins (CKs) of different molecular weights, vascular endothelial cell markers and VEGF/receptor-2 (Flk-1) present on the neovascular endothelial cells. Different points raised by the originality of this model are discussed. These include the medullary nature of the cells expressing the SV40 transgene and their relationship with the tumoral development. The subset of different molecular weight CK components and their modifications are also considered, as well as the presence of type IV epithelial cells, progenitors of medullary epithelial cells. Finally, the cell signals involved in angiogenesis and the possible action of an angiogenic factor, probably secreted by the tumoral cells themselves, are discussed. PMID- 16309546 TI - The haemotoxicity of mitomycin in a repeat dose study in the female CD-1 mouse. AB - Mitomycin (MMC), like many antineoplastic drugs, induces a predictable, dose related, bone marrow depression in man and laboratory animals; this change is generally reversible. However, there is evidence that MMC may also cause a late stage or residual bone marrow injury. The present study in female CD-1 mice investigated the haematological and bone marrow changes induced by MMC in a repeat dose study lasting 50 days. Control and MMC-treated mice were dosed intraperitoneally on eight occasions over 18 days with vehicle, or MMC at 2.5 mg/kg, autopsied (n = 6-12) at 1, 7, 14, 28, 42 and 50 days after the final dose and haematological changes investigated. Femoral nucleated bone marrow cell counts and levels of apoptosis were also evaluated and clonogenic assays carried out; serum levels of FLT3 ligand (FL) were assessed. At day 1 post-dosing, MMC induced significant reductions in RBC, Hb and haematocrit (HCT) values, and there were decreases in reticulocyte, platelet, and femoral nucleated cell counts (FNCC); neutrophil, lymphocyte and monocyte values were also significantly reduced. On days 7 and 14 post-dosing, all haematological parameters showed evidence of a return towards normal values, but at these times, and at day 28, values for RBC and FNCC remained significantly reduced in comparison with controls. At days 42 and 50 post-dosing, many haematological parameters in MMC treated mice had returned to control levels; however, there remained evidence of late-stage effects on RBC, Hb and HCT values, and FNCC also continued to be significantly decreased. Results for granulocyte-macrophage colony-forming units and erythroid colonies showed a profound decrease immediately post-dosing, but a return to normal values was evident at day 50. Serum FL concentrations demonstrated very significant increases in the immediate post-dosing period, but a return to normal was seen at day 50 post-dosing; a relatively similar pattern was seen in the number of apoptotic femoral marrow nucleated cells. The histopathological examination of kidney tissues from MMC animals at day 42 and 50 post-dosing showed evidence of hydronephrosis with cortical glomerular/tubular atrophy and degeneration. It is therefore concluded that MMC administered on eight occasions over 18 days to female CD-1 mice at 2.5 mg/kg induced profound changes in haematological and bone marrow parameters in the immediate post-dosing period with a return to normal levels at day 50 post-dosing; however, there was evidence of mild but significant late-stage/residual effects on RBC and FNCC, and on cells of the erythroid lineage in the bone marrow. PMID- 16309547 TI - Invasive versus noninvasive measurement of allergic and cholinergic airway responsiveness in mice. AB - BACKGROUND: This study seeks to compare the ability of repeatable invasive and noninvasive lung function methods to assess allergen-specific and cholinergic airway responsiveness (AR) in intact, spontaneously breathing BALB/c mice. METHODS: Using noninvasive head-out body plethysmography and the decrease in tidal midexpiratory flow (EF50), we determined early AR (EAR) to inhaled Aspergillus fumigatus antigens in conscious mice. These measurements were paralleled by invasive determination of pulmonary conductance (GL), dynamic compliance (Cdyn) and EF50 in another group of anesthetized, orotracheally intubated mice. RESULTS: With both methods, allergic mice, sensitized and boosted with A. fumigatus, elicited allergen-specific EAR to A. fumigatus (p < 0.05 versus controls). Dose-response studies to aerosolized methacholine (MCh) were performed in the same animals 48 h later, showing that allergic mice relative to controls were distinctly more responsive (p < 0.05) and revealed acute airway inflammation as evidenced from increased eosinophils and lymphocytes in bronchoalveolar lavage. CONCLUSION: We conclude that invasive and noninvasive pulmonary function tests are capable of detecting both allergen-specific and cholinergic AR in intact, allergic mice. The invasive determination of GL and Cdyn is superior in sensitivity, whereas the noninvasive EF50 method is particularly appropriate for quick and repeatable screening of respiratory function in large numbers of conscious mice. PMID- 16309548 TI - Effects of cigarette smoke condensate on proliferation and wound closure of bronchial epithelial cells in vitro: role of glutathione. AB - BACKGROUND: Increased airway epithelial proliferation is frequently observed in smokers. To elucidate the molecular mechanisms leading to these epithelial changes, we studied the effect of cigarette smoke condensate (CSC) on cell proliferation, wound closure and mitogen activated protein kinase (MAPK) activation. We also studied whether modulation of intracellular glutathione/thiol levels could attenuate CSC-induced cell proliferation. METHODS: Cells of the bronchial epithelial cell line NCI-H292 and subcultures of primary bronchial epithelial cells were used for the present study. The effect of CSC on epithelial proliferation was assessed using 5-bromo-2-deoxyuridine (BrdU) incorporation. Modulation of epithelial wound repair was studied by analysis of closure of 3 mm circular scrape wounds during 72 hours of culture. Wound closure was calculated from digital images obtained at 24 h intervals. Activation of mitogen-activated protein kinases was assessed by Western blotting using phospho-specific antibodies. RESULTS: At low concentrations CSC increased proliferation of NCI H292 cells, whereas high concentrations were inhibitory as a result of cytotoxicity. Low concentrations of CSC also increased epithelial wound closure of both NCI-H292 and PBEC, whereas at high concentrations closure was inhibited. At low, mitogenic concentrations, CSC caused persistent activation of ERK1/2, a MAPK involved in cell proliferation. Inhibition of cell proliferation by high concentrations of CSC was associated with activation of the pro-apoptotic MAP kinases p38 and JNK. Modulation of intracellular glutathione (GSH)/thiol levels using N-acetyl-L-cysteine, GSH or buthionine sulphoximine (BSO), demonstrated that both the stimulatory and the inhibitory effects of CSC were regulated in part by intracellular GSH levels. CONCLUSION: These results indicate that CSC may increase cell proliferation and wound closure dependent on the local concentration of cigarette smoke and the anti-oxidant status. These findings are consistent with increased epithelial proliferation in smokers, and may provide further insight in the development of lung cancer. PMID- 16309549 TI - An economic way of reducing health, environmental, and other pressures of urban traffic: a decision analysis on trip aggregation. AB - BACKGROUND: Traffic congestion is rapidly becoming the most important obstacle to urban development. In addition, traffic creates major health, environmental, and economical problems. Nonetheless, automobiles are crucial for the functions of the modern society. Most proposals for sustainable traffic solutions face major political opposition, economical consequences, or technical problems. METHODS: We performed a decision analysis in a poorly studied area, trip aggregation, and studied decisions from the perspective of two different stakeholders, the passenger and society. We modelled the impact and potential of composite traffic, a hypothetical large-scale demand-responsive public transport system for the Helsinki metropolitan area, where a centralised system would collect the information on all trip demands online, would merge the trips with the same origin and destination into public vehicles with eight or four seats, and then would transmit the trip instructions to the passengers' mobile phones. RESULTS: We show here that in an urban area with one million inhabitants, trip aggregation could reduce the health, environmental, and other detrimental impacts of car traffic typically by 50-70%, and if implemented could attract about half of the car passengers, and within a broad operational range would require no public subsidies. CONCLUSION: Composite traffic provides new degrees of freedom in urban decision-making in identifying novel solutions to the problems of urban traffic. PMID- 16309550 TI - Callosal connections of dorsal versus ventral premotor areas in the macaque monkey: a multiple retrograde tracing study. AB - BACKGROUND: The lateral premotor cortex plays a crucial role in visually guided limb movements. It is divided into two main regions, the dorsal (PMd) and ventral (PMv) areas, which are in turn subdivided into functionally and anatomically distinct rostral (PMd-r and PMv-r) and caudal (PMd-c and PMv-c) sub-regions. We analyzed the callosal inputs to these premotor subdivisions following 23 injections of retrograde tracers in eight macaque monkeys. In each monkey, 2-4 distinct tracers were injected in different areas allowing direct comparisons of callosal connectivity in the same brain. RESULTS: Based on large injections covering the entire extent of the corresponding PM area, we found that each area is strongly connected with its counterpart in the opposite hemisphere. Callosal connectivity with the other premotor areas, the primary motor cortex, prefrontal cortex and somatosensory cortex varied from one area to another. The most extensive callosal inputs terminate in PMd-r and PMd-c, with PMd-r strongly connected with prefrontal cortex. Callosal inputs to PMv-c are more extensive than those to PMv-r, whose connections are restricted to its counterpart area. Quantitative analysis of labelled cells confirms these general findings, and allows an assessment of the relative strength of callosal inputs. CONCLUSION: PMd r and PMv-r receive their strongest callosal inputs from their respective counterpart areas, whereas PMd-c and PMv-c receive strong inputs from heterotopic areas as well (namely from PMd-r and PMv-r, respectively). Finally, PMd-r stands out as the lateral premotor area with the strongest inputs from the prefrontal cortex, and only the PMd-c and PMv-c receive weak callosal inputs from M1. PMID- 16309551 TI - Role of rapid urease test and histopathology in the diagnosis of Helicobacter pylori infection in a developing country. AB - BACKGROUND: The aim of this study was to determine the effect of commonly self prescribed proton pump inhibitors (PPI) on the results of rapid urease test and histology for the diagnosis of H. pylori infection. METHODS: One hundred-nine consecutive patients with dyspeptic symptoms attending the endoscopy suite were enrolled in this study. Antrum biopsy specimens were collected at endoscopy for the rapid urease test (Pronto Dry, Medical Instrument Corp, France) and histopathology. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and like-hood ratio of a positive and negative of Pronto Dry test were compared against histology. The gold standard test for the diagnosis of H. pylori infection was histopathology. RESULTS: Sixty-one percent (66/109) patients were males with mean age of 43 +/- 14.1 years and age range 17 80 years. Fifty-two percent (57/109) were not on any medications while 48% (52/109) used PPI before presentation to the outpatients. Pronto Dry was positive in 40% (44/109) and negative in 60% (65/109). Histopathology was positive for H. pylori in 57% (62/109) and negative in 43% (47/109). The sensitivity, specificity, PPV, NPV and like-hood ratio of a positive and negative Pronto Dry test with and without PPI were 43.3%, 86.4%, 81.3%, 3.18, 0.656 and 52.8% vs 71.9%, 80%, 82.1%, 69%, 3.59 and 0.35. CONCLUSION: This study shows that the sensitivity, specificity, NPV and PPV of rapid urease test was reduced in patients who are on PPI. The exclusive use of the rapid urease test for the diagnosis of Helicobacter pylori cannot be recommended in patients with prior PPI use. PMID- 16309552 TI - Nurr1 dependent regulation of pro-inflammatory mediators in immortalised synovial fibroblasts. AB - BACKGROUND: Nurr1 is an orphan member of the nuclear receptor superfamily; these orphan receptors are a group for which a ligand has yet to be identified. Nurr1 has been shown to regulate the expression of a small number of genes as a monomeric, constitutively active receptor. These Nurr1 regulated genes are primarily associated with dopamine cell maturation and survival. However, previous reports have shown an increased expression of Nurr1 in the synovium of patients with rheumatoid arthritis (RA) suggesting a pro-inflammatory role for Nurr1 in RA. In this study we investigate the potential pro-inflammatory role of Nurr1 by monitoring Nurr1 dependent gene expression in an immortalised synoviocyte cell line, K4IM. METHODS: We overexpressed the wild type and a dominant negative form of the orphan nuclear receptor Nurr1, in a model synoviocyte cell line. Using the Affymetrix HG-U133 Genechips we demonstrate the effects on the transcriptome by the receptor. Further evidence of gene expression change was demonstrated using quantitative RT-PCR and ELISA analysis. RESULTS: We show that Nurr1 regulates transcription of a small number of genes for pro inflammatory modulators of which the most significant is interleukin-8 (IL-8). We also demonstrate increased synthesis and secretion of IL-8 further supporting a role for Nurr1 in inflammatory signalling pathways. CONCLUSION: Using microarray analysis we show that elevated levels of Nurr1 leads to increased gene expression of pro-inflammatory genes: IL-8, Amphiregulin and Kit ligand in a model cell line. This data provides further evidence for an additional role for Nurr1 in inflammation and may play a role in the pathogenesis of rheumatoid arthritis. PMID- 16309553 TI - Cardiovascular health knowledge and behavior in patient attendants at four tertiary care hospitals in Pakistan--a cause for concern. AB - BACKGROUND: Knowledge about coronary heart disease (CHD) and its risk factors is an important pre-requisite for an individual to implement behavioral changes leading towards CHD prevention. There is scant data on the status of knowledge about CHD in the general population of Pakistan. The objective of this study was to assess knowledge of CHD in a broad Pakistani population and identify the factors associated with knowledge. METHODS: Cross sectional study was carried out at four tertiary care hospitals in Pakistan using convenience sampling. Standard questionnaire was used to interview 792 patient attendants (persons accompanying patients). Knowledge was computed as a continuous variable based on correct answers to fifteen questions. Multivariable linear regression was conducted to determine the factors independently associated with knowledge. RESULTS: The mean age was 38.1 (+/- 13) years. 27.1% had received no formal education. The median knowledge score was 3.0 out of a possible maximum of 15. Only 14% were able to correctly describe CHD as a condition involving limitation in blood flow to the heart. Majority of respondents could identify only up to two risk factors for CHD. Most commonly identified risk factors were stress (43.4%), dietary fat (39.1%), smoking (31.9%) and lack of exercise (17.4%). About 20% were not able to identify even a single risk factor for CHD. Factors significantly associated with knowledge included age (p = 0.023), income (p < 0.001), education level (p < 0.001), residence (p < 0.001), a family history of CHD (p < 0.001) and a past history of diabetes (p = 0.004). Preventive practices were significantly lacking; 35%, 65.3% and 84.6% had never undergone assessment of blood pressure, glucose or cholesterol respectively. Only a minority felt that they would modify their diet, stop smoking or start exercising if a family member was to develop CHD. CONCLUSION: This is the first study assessing the state of CHD knowledge in a relatively diverse non-patient population in Pakistan. There are striking gaps in knowledge about CHD, its risk factors and symptoms. These translate to inadequate preventive behavior patterns. Educational programs are urgently required to improve the level of understanding of CHD in the Pakistani population. PMID- 16309554 TI - Intravitreal triamcinolone with transpupillary therapy for subfoveal choroidal neovascularization in age related macular degeneration. A randomized controlled pilot study [ISRCTN74123635]. AB - BACKGROUND: To assess the effect of intravitreal triamcinolone acetonide (iTA) as an adjunctive treatment to transpupillary therapy (TTT) for new subfoveal choroidal neovascular membranes (CNV) in age-related macular degeneration (AMD). METHODS: This prospective randomized controlled pilot study comprised 26 patients scheduled to receive TTT, due to either absent indications for photodynamic therapy or financial issues. Patients were assigned into; Group A (n = 14) received TTT alone and Group B (n = 12) received iTA (4 mg) followed by TTT within one week. Follow ups were at 2 weeks, and 1, 3 and 6 months for; best corrected visual acuity (BCVA) by ETDRS chart at 4 meters, intraocular pressures (IOP), fluorescein angiography (FAG), and central foveal thickness by optical coherence tomography (OCT). RESULTS: All 26 patients completed 6 months of follow ups. The average age for both groups was 74 years. Occult CNV formed 64% and 41%; classis/predominately classic 21% and 16.6%; and minimally classic 15% and 42.4% of group A and B respectively. At baseline; the mean BCVA was 0.045 for group A and 0.04 for group B; mean CNV size was 6.15 disc diameter (DD) and 2.44 DD; mean OCT foveal thickness was 513 um and 411 um for group A and B respectively with no statistical differences (P = 0.8, 0.07, and 0.19). At six months the proportion of patients gained > or = 1 lines was 14% and 25% (P = 0.136) and stabilization was 86% and 66% (P = 0.336); the mean size of the CNV was 5.63 DD and 2.67 DD (P = 0.162); rate of CNV closure was 64% and 83% (P = 0.275); and the mean OCT central foveal thickness was 516.36 um and 453.67 um (P = 0.341), for group A and B respectively. CONCLUSION: The use of iTA as an adjunctive to TTT for new subfoveal CNV in AMD showed a tendency towards better functional results. However due to the small sample size of the study a statistically significant results could not be reached. PMID- 16309555 TI - Reduced flavin: NMR investigation of N5-H exchange mechanism, estimation of ionisation constants and assessment of properties as biological catalyst. AB - BACKGROUND: The flavin in its FMN and FAD forms is a versatile cofactor that is involved in catalysis of most disparate types of biological reactions. These include redox reactions such as dehydrogenations, activation of dioxygen, electron transfer, bioluminescence, blue light reception, photobiochemistry (as in photolyases), redox signaling etc. Recently, hitherto unrecognized types of biological reactions have been uncovered that do not involve redox shuffles, and might involve the reduced form of the flavin as a catalyst. The present work addresses properties of reduced flavin relevant in this context. RESULTS: N(5)-H exchange reactions of the flavin reduced form and its pH dependence were studied using the 15N-NMR-signals of 15N-enriched, reduced flavin in the pH range from 5 to 12. The chemical shifts of the N(3) and N(5) resonances are not affected to a relevant extent in this pH range. This contrasts with the multiplicity of the N(5)-resonance, which strongly depends on pH. It is a doublet between pH 8.45 and 10.25 that coalesces into a singlet at lower and higher pH values. From the line width of the 15N(5) signal the pH-dependent rate of hydrogen exchange was deduced. The multiplicity of the 15N(5) signal and the proton exchange rates are little dependent on the buffer system used. CONCLUSION: The exchange rates allow an estimation of the pKa value of N(5)-H deprotonation in reduced flavin to be >or= 20. This value imposes specific constraints for mechanisms of flavoprotein catalysis based on this process. On the other hand the pK asymptotically equal to 4 for N(5)-H protonation (to form N(5)+-H2) would be consistent with a role of N(5)-H as a base. PMID- 16309556 TI - Choosing an appropriate bacterial typing technique for epidemiologic studies. AB - A wide variety of bacterial typing systems are currently in use that vary greatly with respect to the effort required, cost, reliability and ability to discriminate between bacterial strains. No one technique is optimal for all forms of investigation. We discuss the desired level of discrimination and need for a biologic basis for grouping strains of apparently different types when using bacterial typing techniques for different epidemiologic applications: 1) confirming epidemiologic linkage in outbreak investigations, 2) generating hypotheses about epidemiologic relationships between bacterial strains in the absence of epidemiologic information, and 3) describing the distributions of bacterial types and identifying determinants of those distributions. Inferences made from molecular epidemiologic studies of bacteria depend upon both the typing technique selected and the study design used; thus, choice of typing technique is pivotal for increasing our understanding of the pathogenesis and transmission, and eventual disease prevention. PMID- 16309557 TI - Association of common variation in the PPARA gene with incident myocardial infarction in individuals with type 2 diabetes: a Go-DARTS study. AB - BACKGROUND: Common variants of the PPARA gene have been found to associate with ischaemic heart disease in non diabetic men. The L162V variant was found to be protective while the C2528G variant increased risk. L162V has also been associated with altered lipid measures. We therefore sought to determine the effect of PPARA gene variation on susceptibility to myocardial infarction in patients with type 2 diabetes. 1810 subjects with type 2 diabetes from the prospective Go-DARTS study were genotyped for the L162V and C2528G variants in the PPARA gene and the association of the variants with incident non-fatal myocardial infarction was examined. Cox's proportional hazards was used to interrogate time to event from recruitment, and linear regression for analysing association of genotype with quantitative clinical traits. RESULTS: The V162 allele was associated with decreased risk of non-fatal myocardial infarction (HR = 0.31, 95%CI 0.10-0.93 p = 0.037) whereas the C2528 allele was associated with increased risk (HR = 2.77 95%CI 1.34-5.75 p = 0.006). Similarly V162 was associated with a later mean age of diagnosis with type 2 diabetes and C2582 an earlier age of diagnosis. C2528 was also associated with increased total cholesterol and LDL cholesterol, which did not account for the observed increased risk. Haplotype analysis demonstrated that when both rare variants occurred on the same haplotype the effect of each was abrogated. CONCLUSION: Genetic variation at the PPARA locus is important in determining cardiovascular risk in both male and female patients with diabetes. This genotype associated risk appears to be independent of the effect of these genotypes on lipid profiles and age of diagnosis with diabetes. PMID- 16309558 TI - Suppression of AC railway power-line interference in ECG signals recorded by public access defibrillators. AB - BACKGROUND: Public access defibrillators (PADs) are now available for more efficient and rapid treatment of out-of-hospital sudden cardiac arrest. PADs are used normally by untrained people on the streets and in sports centers, airports, and other public areas. Therefore, automated detection of ventricular fibrillation, or its exclusion, is of high importance. A special case exists at railway stations, where electric power-line frequency interference is significant. Many countries, especially in Europe, use 16.7 Hz AC power, which introduces high level frequency-varying interference that may compromise fibrillation detection. METHOD: Moving signal averaging is often used for 50/60 Hz interference suppression if its effect on the ECG spectrum has little importance (no morphological analysis is performed). This approach may be also applied to the railway situation, if the interference frequency is continuously detected so as to synchronize the analog-to-digital conversion (ADC) for introducing variable inter-sample intervals. A better solution consists of rated ADC, software frequency measuring, internal irregular re-sampling according to the interference frequency, and a moving average over a constant sample number, followed by regular back re-sampling. RESULTS: The proposed method leads to a total railway interference cancellation, together with suppression of inherent noise, while the peak amplitudes of some sharp complexes are reduced. This reduction has negligible effect on accurate fibrillation detection. CONCLUSION: The method is developed in the MATLAB environment and represents a useful tool for real time railway interference suppression. PMID- 16309559 TI - Genes involved in complex adaptive processes tend to have highly conserved upstream regions in mammalian genomes. AB - BACKGROUND: Recent advances in genome sequencing suggest a remarkable conservation in gene content of mammalian organisms. The similarity in gene repertoire present in different organisms has increased interest in studying regulatory mechanisms of gene expression aimed at elucidating the differences in phenotypes. In particular, a proximal promoter region contains a large number of regulatory elements that control the expression of its downstream gene. Although many studies have focused on identification of these elements, a broader picture on the complexity of transcriptional regulation of different biological processes has not been addressed in mammals. The regulatory complexity may strongly correlate with gene function, as different evolutionary forces must act on the regulatory systems under different biological conditions. We investigate this hypothesis by comparing the conservation of promoters upstream of genes classified in different functional categories. RESULTS: By conducting a rank correlation analysis between functional annotation and upstream sequence alignment scores obtained by human-mouse and human-dog comparison, we found a significantly greater conservation of the upstream sequence of genes involved in development, cell communication, neural functions and signaling processes than those involved in more basic processes shared with unicellular organisms such as metabolism and ribosomal function. This observation persists after controlling for G+C content. Considering conservation as a functional signature, we hypothesize a higher density of cis-regulatory elements upstream of genes participating in complex and adaptive processes. CONCLUSION: We identified a class of functions that are associated with either high or low promoter conservation in mammals. We detected a significant tendency that points to complex and adaptive processes were associated with higher promoter conservation, despite the fact that they have emerged relatively recently during evolution. We described and contrasted several hypotheses that provide a deeper insight into how transcriptional complexity might have been emerged during evolution. PMID- 16309560 TI - The EH1 motif in metazoan transcription factors. AB - BACKGROUND: The Engrailed Homology 1 (EH1) motif is a small region, believed to have evolved convergently in homeobox and forkhead containing proteins, that interacts with the Drosophila protein groucho (C. elegans unc-37, Human Transducin-like Enhancers of Split). The small size of the motif makes its reliable identification by computational means difficult. I have systematically searched the predicted proteomes of Drosophila, C. elegans and human for further instances of the motif. RESULTS: Using motif identification methods and database searching techniques, I delimit which homeobox and forkhead domain containing proteins also have likely EH1 motifs. I show that despite low database search scores, there is a significant association of the motif with transcription factor function. I further show that likely EH1 motifs are found in combination with T Box, Zinc Finger and Doublesex domains as well as discussing other plausible candidate associations. I identify strong candidate EH1 motifs in basal metazoan phyla. CONCLUSION: Candidate EH1 motifs exist in combination with a variety of transcription factor domains, suggesting that these proteins have repressor functions. The distribution of the EH1 motif is suggestive of convergent evolution, although in many cases, the motif has been conserved throughout bilaterian orthologs. Groucho mediated repression was established prior to the evolution of bilateria. PMID- 16309561 TI - The variable number of tandem repeats element in DAT1 regulates in vitro dopamine transporter density. AB - BACKGROUND: A 40-bp variable number of tandem repeats (VNTR) polymorphism exists in the 15th exon of DAT1, the gene encoding the human dopamine transporter (DAT). Though the VNTR resides in a region encoding the 3' untranslated region (UTR) and does not alter the protein's amino acid sequence, the prevalent 10-repeat variant has shown both linkage and association to Attention Deficit Hyperactivity Disorder (ADHD). In this study, we examined the effects of the DAT1 VNTR on measures of in vitro DAT expression and pharmacology. A series of four DAT1 constructs, each containing the DAT1 coding region, but varying with respect to the downstream presence or content of the 3'UTR, were engineered and stably transfected into an HEK-293 variant using Flp-In integration, an enzyme-mediated, site-specific recombination technology. RESULTS: [3H] Win 35,428 saturation binding assays and DAT immunoblots revealed statistically significant differences in DAT expression attributable to DAT1 genotype. Cells harboring the 10-repeat DAT1 variant were characterized by a Bmax approximately 50% greater than cells with the 9-repeat VNTR; those containing only the DAT1 coding region or the coding region flanked by a truncated 3' UTR resulted in greater DAT density than either of the naturalistic 9- and 10-repeat variants. Competition binding assays showed no statistically significant DAT1 genotype effects on the DAT affinity for methylphenidate, a finding consistent with the positional location of the VNTR. CONCLUSION: This study identified the DAT1 VNTR as a functional polymorphism and provides an interpretive framework for its association with behavioral phenotypes. PMID- 16309562 TI - Unusual explosive growth of a squamous cell carcinoma of the scalp after electrical burn injury and subsequent coverage by sequential free flap vascular connection--a case report. AB - BACKGROUND: Squamous cell carcinomos may arise from chronic ulcerating wounds in scars, most commonly postburn scars. Tumour growth usually takes place over months to years. Localization on the scalp is a relatively rare condition. CASE PRESENTATION: This report presents the case of a 63-year-old man with chronic ulceration of a postburn scar of the scalp due to an electrical burn 58 years ago. Sudden tumour growth started within weeks and on presentation already had extended through the skull into frontal cortex. After radical tumour resection, defect was covered with a free radial forearm flap. Local recurrence occurred 6 weeks later. Subsequent wide excision including discard of the flap and preservation of the radial vessels was followed by transfer of a free latissimus dorsi muscle flap, using the radial vessels of the first flap as recipient vessels. The patient received radiotherapy post-operatively. There were no problems with flap survivals or wound healing. Due to rapidly growing recurrence the patient died 2 months later. CONCLUSION: Explosive SCC tumour growth might occur in post-burn scars after more than 50 years. As a treatment option the use of sequential free flap connections might serve in repeated extensive tumour resections, especially in the scalp region, where suitable donor vessels are often located in distance to the defect. PMID- 16309563 TI - Fine structure of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 (Monogenea: Ancyrocephalidae). AB - A study of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 was carried out using light and electron microscopy. The anterior adhesive apparatus or head organs in B. gussevi comprise 6 circular openings or apertures in the antero-lateral region, associated pits lined with specialized microvillous tegument that differ from the general body tegument, a bundle of ducts, and uninucleate gland cells located lateral to the pharynx. The uninucleate glands of the anterior adhesive apparatus (head organs) comprise 2 types of cells, one kind of cell producing rod-like bodies (S1) and the other oval bodies (S2). The S1 bodies are filled with numerous, less electron-dense vesicles in an electron-dense matrix, while S2 bodies have no vesicles but contain a more homogeneous electron-dense matrix. Interlinking band-like structures were observed between S1 bodies. Similar band-like structures were found between S2 bodies. The formation of S1 bodies was followed by transmission electron microscopy. However, the formation of S2 bodies was unclear and could not be resolved. Uniciliated structures were also observed around the openings of the anterior adhesive apparatus. Each uniciliated structure is usually associated with an opening of a gland cell producing granular, electron-dense, secretory bodies, which differ from the secretions produced by the lateral gland cells of the anterior adhesive apparatus. PMID- 16309564 TI - Carbon monoxide increases macrophage bacterial clearance through Toll-like receptor (TLR)4 expression. AB - Carbon monoxide (CO), a catabolic product of heme degradation, is an efficacious cytoprotectant and potent anti-inflammatory molecule. One of the important cellular targets of carbon monoxide is the macrophage, a key modulator of inflammation. In this study we investigated the effects of CO on the ability of cultured macrophages to phagocytose E. coli. Exposure to CO augmented E. coli phagocytosis but had no effect on inert particulate matter internalization. The ability of CO to increase uptake of the bacteria was in part mediated by the redistribution and increased expression of Toll-like receptor 4 (TLR4) on the cell surface. Furthermore, inhibition of p38 MAPK attenuated CO/E. coli-induced surface expression of TLR4 and abrogated the CO effects on E. coli phagocytosis. Collectively these data show that CO enhances the rate of E. coli phagocytosis via p38-mediated surface expression of TLR4 and suggest that CO may be a potential therapeutic modality by which to increase bacterial clearance. PMID- 16309565 TI - Local administration of carbon monoxide inhibits neointima formation in balloon injured rat carotid arteries. AB - Recent studies indicate that systemic induction of heme oxygenase-1 (HO-1), which oxidatively degrades heme into iron, biliverdin, and carbon monoxide (CO), or adenoviral-mediated gene transfer of HO-1 inhibits neointima formation after experimental vascular injury. In the present study, we investigated whether the acute, local administration of the HO-1 product, CO, regulates the arterial remodeling response following injury. Immediately after balloon injury of rat carotid arteries, a saturated solution of CO or nitrogen (N2), or phosphate buffered saline (PBS) was incubated luminally within the injured vessels for 30 min. Two weeks after injury, arteries exposed to CO exhibited significantly reduced neointimal area, neointimal area/medial wall area ratio, neointimal thickness, and medial wall area compared to arteries exposed to N2 or PBS. Arteries exposed to CO also demonstrated significantly reduced DNA synthesis in the medial wall two days after injury as suggested by proliferating cell nuclear antigen immunostaining, and this was associated with a decrease in the protein expression of the G1 cyclins, cyclin E and A, and transforming growth factor beta1. These results indicate that the acute, local delivery of CO blocks the pathophysiological remodeling response to vascular injury, and identifies CO as a potentially important therapeutic agent in the treatment of vasculoproliferative disease. PMID- 16309566 TI - The source of endogenous carbon monoxide formation in human placental chorionic villi. AB - Carbon monoxide (CO) is proposed to play a role in placental vascular control, as the placenta produces and responds to CO. The mechanism by which CO is formed by the placenta is unclear but could be through heme oxygenase (HO) degradation of heme, lipid peroxidation, or both. Human placental cotyledons were perfused with Kreb s solution to remove blood. Chorionic villi segments were prepared for measurements of CO production in the absence/presence of an exogenous supply of heme substrate (methemalbumin), inhibitors of HO, or inhibitors of lipid peroxidation. HO inhibitors used were chromium mesoporphyrin (CrMP) (0.1 mM, 0.3 mM), and azalanstat (0.1 mM, 0.3 mM). The lipid peroxidation inhibitors used were EDTA (0.1 mM, 0.3 mM) and deferoxamine (0.1 mM). Incubation of villi segments with methemalbumin (0.15 mM, 0.3 mM, 0.45 mM) resulted in a concentration dependent increase in CO formation above the basal, endogenous rate. CrMP and azalanstat inhibited basal endogenous CO production, whereas EDTA and deferoxamine enhanced CO formation above basal level. These results demonstrate that endogenous CO was formed by human chorionic villi from heme, primarily through the action of HO, and are consistent with the hypothesis that HO plays a role in the regulation of placental vasculature by the formation of heme-derived CO. PMID- 16309567 TI - The role of exogenous carbon monoxide in the recovery of post-ischemic cardiac function in buffer perfused isolated rat hearts. AB - Isolated rat hearts were perfused for 10 min with oxygenated buffer and equilibrated with carbon monoxide (CO) of 0.001% and 0.01% before the induction of 30 min global ischemia followed by 120 min of reperfusion. These concentrations of CO significantly improved the post-ischemic recovery of coronary flow (CF), aortic flow (AF), and left ventricular developed pressure (LVDP). The improvement in recovery reflected in the reduction of infarct size and the incidence of reperfusion-induced ventricular fibrillation (VF). Thus, hearts subjected to 0.001% and 0.01% of CO exposure via the perfusion buffer, infarct size was reduced from the CO-free control value of 39% +/- 5% to 21% +/- 3% (*p<0.05) and 18% +/- 4% (*p<0.05), respectively. In the presence of 0.001% and 0.01% CO, the incidence of VF was also reduced from its control value of 92% to 17% (*p<0.05) and 17% (*p<0.05), respectively. Increasing the CO exposure to 0.1% in the buffer, all hearts showed VF combined with ventricular tachycardia or bradycardia and various rhythm disturbances indicating the direct toxic effects of CO on the myocardium. The results show that cardioprotective concentrations (0.01% and 0.001%) of exogenous CO related to an increase in cGMP levels and guanylate cyclase activities. PMID- 16309568 TI - Phosphatidylinositol 3-kinase/Akt pathway mediates heme oxygenase-1 regulation by lipopolysaccharide. AB - The stress-inducible protein heme oxygenase-1 exerts potent antiinflammatory, antiapoptotic and cytoprotective effects in vitro and in vivo. Another important mediator of cytoprotection, the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway activates many proteins involved in the maintenance of cellular homeostasis. Since activation of heme oxygenase-1 and PI3K/Akt both protect the cellular environment, we postulated that PI3K/Akt can regulate the induction of heme oxygenase-1 by proinflammatory stress. The treatment of primary murine macrophage cells (RAW 264.7) with lipopolysaccharide induced heme oxygenase-1 protein and mRNA expression, and increased the phosphorylation of Akt and p38 mitogen activated protein kinase (p38 MAPK). These cellular effects of lipopolysaccharide were markedly diminished by pre-treatment with wortmannin, a specific inhibitor of PI3K. Furthermore, lipopolysaccharide-inducible heme oxygenase expression was blocked by SB203580, a specific inhibitor of p38 MAPK. Both wortmannin and SB203580 decreased lipopolysaccharide-inducible NF-E2-related factor (Nrf2) DNA binding activity. Transfection of macrophages with dominant negative mutants of PI3K, Akt and Nrf2, as well as wortmannin treatment, significantly reduced the transcriptional activity of a minimal heme oxygenase-1 promoter luciferase construct (D33HO-1luc). We demonstrate, to our knowledge for the first time, that upon proinflammatory stimulation heme oxygenase-1 gene expression in macrophages depends on PI3K/Akt and p38 MAPK acting upstream of Nrf2-dependent promoter activation. PMID- 16309569 TI - Complementary regulation of heme oxygenase-1 and peroxiredoxin I gene expression by oxidative stress in the liver. AB - Heme oxygenase (HO)-1, the inducible isoform of the rate-limiting enzyme of heme degradation, and peroxiredoxin (Prx) I, a thioredoxin-dependent peroxidase, are multifunctional antioxidant stress proteins which are coordinately up-regulated by oxidative stress in cell cultures. HO-1 and Prx I exhibit a different hepatic cellular and subcellular localization. Here, a distinct expression pattern of the two genes was confirmed by in situ hybridization of normal rat liver. Moreover, expression of the HO-1 and Prx I genes was determined in a model of acutely damaged rat liver which was elicited by application of a single dose of carbon tetrachloride (CCl4). The mRNA levels of the HO-1 and Prx I genes were induced in whole livers of CCl4-treated rats with differential kinetics as determined by Northern blot analysis. While HO-1 mRNA was induced up to 48 hr, Prx I exhibited a maximum level of mRNA after 12 hr of treatment with CCl4. CCl4-dependent oxidative stress led to a focal increase of perivenous HO-1 positive liver cells with simultaneous loss of Prx I immunoreactivity. Taken together, the complementary hepatic gene expression pattern of HO-1 and Prx I in response to oxidative stress may suggest a functional interplay of these antioxidant genes. PMID- 16309570 TI - Effects of heme oxygenase-1 inducers on established rat adjuvant arthritis. AB - Heme oxygenase-1 (HO-1) activity can inhibit inflammatory and immune responses. We have examined the influence of HO-1 induction on the established rat adjuvant arthritis model of chronic inflammation. Therapeutic administration of cobalt protoporphyrin IX (CoPP; 5 mg/kg/day i.p.) from day 17 to 23 significantly reduced the inflammatory response, with partial inhibition of hind paw edema and the production of some inflammatory mediators such as nitric oxide metabolites and tumor necrosis factor-alpha, although joint erosion was observed. Hemin administration (26 mg/kg/day i.p.) during the same time period was ineffective on these parameters. Western blot analysis of hind paw homogenates revealed a weaker induction of HO-1 by this compound in comparison with CoPP. Our data indicate that pharmacological HO-1 induction after the establishment of adjuvant arthritis partially reduced the inflammatory response but it was not sufficient to control joint erosion in our experimental conditions. PMID- 16309571 TI - Heme oxygenase, aminolevulinate acid synthetase and the antioxidant system in the brain of mice treated with porphyrinogenic drugs. AB - Several drugs and stress are involved in the triggering of attacks in acute porphyrias. The central nervous system is extremely sensitive to free radical damage because of a relatively low antioxidant capacity. We have demonstrated that mice brain cholinergic system was altered by the effect of some porphyrinogenic agents. The aim of this work was to investigate how known porphyrinogenic drugs affect delta-Aminolevulinic acid synthetase (ALA-S), which is the response of heme oxygenase (HO) to this challenge and to evaluate if the xenobiotics studied develop stress oxidative in mice brain. HO activity was 50 70% induced after chronic Enflurane and Isoflurane anaesthesia, dietary Griseofulvin and starvation. An increase in mRNA HO expression was caused by chronic anaesthesia and Veronal treatments; instead allylisopropilacetamide (AIA) reduced mRNA expression. ALA-S activity was induced by acute administration of anaesthetics (89%), veronal (240%) and ethanol (80%), while ALA-S mRNA expression augmented by chronic administration of enflurane, AIA and veronal. Stress markers such as superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activities and malondialdehyde and reduced glutathione levels showed different responses depending on the xenobiotic assayed. In conclusion, some of the drugs studied produced oxidative stress in brain that was confirmed through HO induction and this could be one of the factors leading to porphyric neuropathy. PMID- 16309572 TI - HO-1 induction in the guinea-pig stomach: protection of smooth muscle functional performance during cobalt-induced oxidative stress. AB - Distribution and enzymatic activity of haem oxygenase (HO) was investigated in the stomach of healthy guinea pigs or animals subjected to in vivo cobalt-induced oxidative stress. The physiological role of HO-1 and HO-2 isoenzymes in fundic and antral area of the stomach was assessed by studying the action of HO substrate--hemin--on ionic currents of single smooth muscle cells. The data obtained suggest that HO-1 induction might serve in the guinea-pig stomach as genetically determined defense mechanism aimed to combat toxic stress-related pathology in order to preserve the functional performance of the organ. PMID- 16309573 TI - Monitoring circulatory heme level in hemin therapy for lowering blood pressure in rats. AB - Hemin and other heme derivatives, e.g. heme-L-lysinate (HLL) and heme-L-arginate, have been used extensively to upregulate expression of heme oxygenase and production of endogenous carbon monoxide. Hemin administration has been shown to markedly decrease high blood pressure in spontaneously hypertensive rats (SHR), but not in normotensive Wistar-Kyoto or Sprague Dawley (SD) rats. While methodology to measure serum heme levels has been established long ago, metabolism of the injected hemin or heme derivatives when used to lower blood pressure has not been investigated. In this study, metabolism of hemin or HLL after injected into the rat was monitored by measuring changes in circulatory heme levels. SHR (12-20 weeks old) had significantly higher blood pressure than age-matched SD rats. In both strains, serum heme level was negligible. Hemin or HLL injection (15 mg/kg/day, i.p.) for 5-13 days significantly lowered blood pressure of 12-weeks SHR. High blood pressure was not lowered in SHR older than 20 weeks until hemin or HLL injection period was beyond 5 days. This anti hypertensive effect of hemin and HLL was synchronized with an increase in serum heme level, from undetectable to 4.3 micromol/l. On the other hand, hemin or HLL had no effect on blood pressure of age-matched SD rats, despite serum heme level rose to the same extent as in the treated SHR. There was no significant difference between hemin and HLL injections in terms of changes in blood pressure and serum heme level in all rats. Our study for the first time correlated changes in serum heme levels with blood pressure levels after injection of hemin or HLL in SHR and SD rats. Hemin and HLL had similar effects on blood pressure change and serum heme level. By determining serum heme levels following the administration of hemin or HLL, we can better understand mechanisms for the blood pressure lowering effect of hemin therapy. Application of this heme monitoring technology will also pave the way for clinical application of hemin therapy in treatment of different types of hypertension pathologies. PMID- 16309574 TI - Absence of heme oxygenase-1 expression in the lung parenchyma exacerbates endotoxin-induced acute lung injury and decreases surfactant protein-B levels. AB - Acute respiratory distress syndrome (ARDS) is a devastating disease process characterized by severe acute lung injury, inflammatory cell recruitment to the lung, upregulation of pro-inflammatory cytokines and increased oxidative stress. Epithelial cell injury, diffuse alveolar damage and surfactant dysfunction ensue leading to refractory hypoxemic respiratory failure. There are no specific effective therapies for ARDS and novel therapeutic approaches are desperately needed. In this study we assessed the role of the cytoprotective and anti inflammatory enzyme heme oxygenase (HO)-1 in a model of nebulized endotoxin induced acute lung injury. HO-1 null (HO-1(-/-)) mice exhibited severe physiologic lung dysfunction following lipopolysaccharide (LPS) nebulization, but had similar inflammatory responses as wild-type (WT) mice. However, a dramatic reduction in surfactant protein-B (SP-B) expression was observed in the lungs of LPS-treated HO-1(-/-) mice compared with similarly treated WT mice. Using reciprocal bone marrow transplantation (BMT) to generate HO-1-chimeric mice, we found that absence of HO-1 in the lung parenchyma, not in bone marrow-derived inflammatory cells, was responsible for enhanced SP-B downregulation and severe physiologic lung dysfunction. These findings have implications for our understanding of the pathophysiology of ARDS and may guide future therapeutic strategies. PMID- 16309575 TI - Antihypertensive pharmacological agents: beyond their lowering blood pressure effects. PMID- 16309576 TI - Dehydroleucodine inhibits vascular smooth muscle cell proliferation in G2 phase. AB - Vascular smooth muscle cell (VSMC) proliferation plays an important role in the development of atherosclerosis and in the vascular changes seen in hypertension. Dehydroleucodine (DhL) is a sesquiterpene lactone that inhibits cell proliferation in plant cells. In this paper, we study the effect of DhL in the proliferation of VSMCs stimulated with 10% fetal bovine serum (FBS). Very low concentrations of DhL (2-6 microM) inhibited VSMC proliferation and induced cell accumulation in G2. DhL did not affect the dynamics of 3H-thymidine incorporation, and did not modify either the activity of DNA polymerase or the incorporation of deoxyribonucleotides in an in vitro assay. Moreover, DhL did not induce apoptosis in VSMCs. These results indicate that DhL, in very low concentration, induces a transient arrest of VSMCs in G2. Our data show that VSMCs are especially sensitive to DhL effect, suggesting that DhL could be potentially useful to prevent the vascular pathological changes seen in hypertension and other vascular diseases. PMID- 16309577 TI - Effect of nebivolol on cardiovascular changes associated with a rat model of insulin-resistance. AB - Nebivolol is a vasodilator that combines beta-adrenergic blocking activity with a relaxant effect on vascular smooth muscle cells (VSMC) mediated by the endothelial nitric oxide (NO) pathway. FFR provide a model of dietary-induced insulin-resistance syndrome, which has been used to study the pathophysiological mechanisms associated with this syndrome. Our main objective was to examine the effect of long-term administration of nebivolol on metabolic and cardiovascular variables in fructose-fed rats (FFR), a model in which an altered bioavailability of NO has been already described. Male Wistar rats were randomly assigned to 4 groups (n = 8 each): I. Control (C); II. Control + nebivolol (C+N): 1 mg/kg(-1) x day(-1) in drinking water during the last 4 weeks. III. FFR: rats receiving fructose in drinking water as a 10% (w/v) solution during 8 weeks, and IV. FFR+N: idem II plus III. During the 8 weeks experimental period, variations in systolic blood pressure (SBP), glucose tolerance test (GTT) and plasma thiobarbituric acid reactive substances (TBARS) were assessed. At the end of this experimental period, rats were killed and heart and kidneys were excised for calculation of relative heart weight (RHW) and histological evaluation of lumen to media ratio (L/M) in renal arteries. Rats from FFR group increased their SBP and RHW, showed glucose intolerance and an increment in lipid peroxidation. Moreover, FFR showed vascular remodeling in renal arteries evidenced by changes in L/M. Although the metabolic changes were not reverted by the administration of nebivolol, this drug successfully decreased SBP, TBARS levels and reverted structural changes such as cardiac hypertrophy and renal arterial remodeling. Data demonstrate that nebivolol administration could participate in the reversion of cardiovascular structural changes associated with the insulin-resistance syndrome. PMID- 16309578 TI - Effect of losartan pretreatment on kidney lipid content after unilateral obstruction in rats. AB - Intrarenal concentration of angiotensin II increases after the onset of ureteral obstruction in the obstructed kidney. The effect of pretreatment with losartan, a specific angiotensin II AT1 receptor antagonist, on lipid contents, which were previously modified by unilateral ureteral obstruction (UUO), was studied in renal cortex of rats. Adult Wistar Kyoto rats were subjected to either UUO for 24 hr or control sham operation after being treated with losartan in the drinking water at 10 mg/kg rat/day for 15 days. In the cortex of obstructed kidney the increased free and esterified cholesterol concentrations were associated with the increased cholesterol synthesis measured by incorporation of 14C-acetate (0.001>p), compared with control and contralateral kidneys. The increased amount of phosphatidylcholine was related with the increased incorporation of 14C choline into phosphatidylcholine (0.01>p). Phosphatidylethanolamine and sphingomyelin decreased slightly but total phospholipid content did not change. The level of AT1 receptor mRNA in obstructed kidney was significantly lower than in control and contralateral kidneys. Losartan pretreatment attenuated (0.01>p) the increase in cholesterol content and synthesis and restored and enhanced the AT1 angiotensin II receptor gene expression. The interference in the renin angiotensin system before UUO may modify renal cortex cholesterol content. PMID- 16309579 TI - Angiotensin II type I antagonist on oxidative stress and heat shock protein 70 (HSP 70) expression in obstructive nephropathy. AB - Angiotensin II, a profibrotic cytokine, plays a main role in the initiation of renal fibrogenesis at a very early stage leading to a progressive loss of renal function in unilateral ureteral obstruction (UUO). We studied the involvement of AT1 angiotensin II receptor in the physiopathology of tubulointerstitial fibrosis in UUO, focusing in the regulation of the oxidative stress state and in the HSP 70 expression, in renal tissue. UUO or control sham operation was perform to Wistar Kyoto rats after being treated with the AT1 angiotensin II receptor antagonist Losartan (10 mg/kg/day) in the drinking water for 15 days. Twenty four hours later, mRNA AT1 receptor expression was studied. Renal fibrosis was evaluated through TGFbeta expression and superoxide dismutase (SOD) activity, hydroxyl radicals, O2- and total antioxidant activity were measured by spectrophotometric assay. Immunohistochemical and Western blot analysis of HSP 70 were performed. A non-hypotensive dose of Losartan significantly down regulated the expression of AT1 receptor. Prevention of renal fibrogenesis by Losartan treatment was demonstrated by TGFbeta mRNA expression similar to control. Oxidative stress in obstructed kidney was evident since a decreased SOD activity and a two-fold increase in the concentration of hydroxyl radicals and O2- was observed when compared to the control. Losartan produced down regulation of ROS with recovery of the SOD activity and higher expression of HSP 70 compared to obstructed kidney of rats receiving vehicle. We can conclude that after 24 hr of UUO, protection against tubulointerstitial fibrosis by Losartan, independent from changes in blood pressure, includes decreased oxidative stress linked to upregulation of HSP 70 expression. PMID- 16309580 TI - Effect of omapatrilat on the aging process of the normal rat. AB - Previous results have shown that inhibition of the renin-angiotensin system (RAS) either with an angiotensin II (Ang II), type 1 receptor blocker (losartan) or with an angiotensin converting enzyme inhibitor (ACEI, enalapril) has a protective effect on cardiovascular, renal, hepatic and cerebral structure and function during aging. The present study has analyzed the effect of chronic administration of a newly developed compound, omapatrilat, on clinical, histological and biochemical changes due to aging. Omapatrilat combines the action of an ACEI and of an inhibitor of a neutral endopeptidase involved in the metabolism of the atrial natriuretic peptide. The final effect is a decrease of a vasoconstrictor and proinflammatory mechanism like the RAS and the potentiation of two vasodilating compounds like bradykinin and the atrial natriuretic peptide. Based on these actions, its protective effect might be greater than formerly used pharmacological agents. Determinations have been performed on young adults (6 months old), adults (12 months old) or senile (18 months old) rats. Omapatrilat (35 mg/kg/day during 6 months and 20 mg/kg/day thereafter) was administered in the drinking water since weaning until sacrifice. Cardiovascular, renal, and cerebral structure as well as cognitive behavior, cardiovascular and renal function has been analyzed. The biochemical analysis has also established whether the beneficial action of Ang II inhibition is related to an increased activity of the nitric oxide synthase as observed in previous studies. Moreover, this study has tried to determine the relationship between the protective effect of these drugs and the levels of antioxidant defenses present in the blood and/or in the tissues. Hence, enzymatic and non-enzymatic antioxidants have been evaluated. PMID- 16309581 TI - Insulin improves the endothelium-independent relaxation and the contractile response in aorta from hypertensive diabetic rats. AB - We investigated the effect of insulin on basal tone and contractile response in isolated aorta from hypertensive streptozotocin-induced diabetic rats (DR) and the role of endothelium in this response. The effect of insulin was tested in rings of control rats (CR) and DR in different protocols: in basal tone, in the plateau of norepinephrine (NE) or KCl contracted rings and in the response to NE or KCl preincubated with insulin. The role of nitric oxide (NO) on insulin response was investigated in rings treated with L-NAME. We found in DR: a) An endothelium independent-vasorelaxant effect of insulin on basal tone; b) A decreased response to NE (without differences in the sensibility) and to KCl (20 mmol/l) and an improvement of this hyporeactivity by insulin pre-treatment, and c) A potentiated vasorelaxant response of insulin dependent of increased vascular tone. Furthermore, an additional action of insulin on endothelial response through NO-release was observed in precontracted vessels from CR, not observed in DR. Our results support that insulin plays a role in regulation of arterial basal tone from DR by a direct effect on smooth muscle vascular cells exposed to high blood pressure. The vasorelaxant effect of insulin dependent of endothelium is blunted in DR by a reduced endothelial NO production. Our work also suggest that insulin could improve the endothelial function in vessels with increased tone in absence of diabetes. PMID- 16309582 TI - Angiotensin II blockade improves mitochondrial function in spontaneously hypertensive rats. AB - Angiotensin II can induce oxidant stress by stimulating vascular superoxide production. Hypertension promotes mitochondrial function decline in brain, liver and heart. The aim of this study was to investigate whether a) hypertension is associated to kidney mitochondrial dysfunction, and b) angiotensin II blockade can reverse potential mitochondrial changes in hypertension. Four-month-old male spontaneously hypertensive rats (SHR) received drinking water containing candesartan (7.5 mg/kg/day, SHR+Cand), or no additions (SHR) for 4-months. Eight month-old Wistar-Kyoto rats (WKY), that received water with no additions, were used as control. Systolic blood pressure, proteinuria, cortical glomerular area, and glomerular and tubulointerstitial alpha-smooth muscle actin labeling, were significantly higher, and creatinine clearance was significantly lower, in SHR relative to WKY and SHR+Cand. In SHR, kidney mitochondria membrane potential, and nitric oxide synthase and cytochrome oxidase activities were significantly lower than in WKY and SHR+Cand. In SHR, mitochondrial hydrogen peroxide production was significantly higher than in WKY and SHR+Cand. The results suggest that, in hypertension, increased mitochondrial oxidant production may mediate kidney mitochondria dysfunction. Candesartan preserved mitochondrial function, probably favoring the maintenance of adequate cellular and tissue function in the kidney. The known renal protective effects of candesartan in hypertension may be related to the improvement of mitochondrial function. This may be an additional or alternative explanation for some of the beneficial effects of AT1 receptor antagonists. PMID- 16309583 TI - Heme oxygenase-1: a key step in counteracting cellular dysfunction. PMID- 16309584 TI - Heme oxygenase-1-dependent and -independent regulation of angiogenic genes expression: effect of cobalt protoporphyrin and cobalt chloride on VEGF and IL-8 synthesis in human microvascular endothelial cells. AB - Induction of heme oxygenase-1 (HO-1) expression can be achieved by stimulation with cobalt protoporphyrin (CoPPIX) or cobalt chloride (CoCl2). HO-1 has been recently implicated in regulation of angiogenesis and CoCl2 is known to potently activate hypoxia inducible factor-1 (HIF-1) transcription factor, a key regulator of angiogenic response in hypoxia. Here we determined the effect of CoPPIX and CoCl2 on the expression of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8), the two major angiogenic mediators, in human microvascular endothelial cells (HMEC-1). CoPPIX induced HO-1 expression and strongly enhanced VEGF and IL-8 synthesis, through the activation of VEGF and IL-8 promoters. Inhibition of HO activity by SnPPIX decreased VEGF production, while, interestingly, it did not affect IL-8. CoCl2 activated hypoxia-responsive element (HRE) and consequently VEGF generation via the enhancement of production of reactive oxygen species (ROS). On the other hand, CoCl2 did not influence IL-8 expression, while CoPPIX did not induce ROS elevation neither it affected HRE activity in VEGF promoter. Our data show that although both CoCl2 and CoPPIX induce HO-1, the influence of CoCl2 on VEGF does not involve HO-1 and is HIF-1 dependent, while the effect of CoPPIX does not involve HIF-1 but relies on HO-1. PMID- 16309585 TI - Induction of heme oxygenase-1 modulates the profibrotic effects of transforming growth factor-beta in human renal tubular epithelial cells. AB - Transforming growth factor-beta (TGF-beta) is implicated in a variety of kidney diseases where it promotes extracellular matrix (ECM) deposition and pro inflammatory events, but it also stabilizes and attenuates tissue injury through the activation of cytoprotective proteins, including heme oxygenase-1 (HO-1). HO 1 catalyzes the conversion of heme into carbon monoxide (CO), iron, and biliverdin, which is subsequently converted to bilirubin. The beneficial effects of HO-1 induction include decreasing pro-oxidants (heme), increasing anti oxidants (biliverdin and bilirubin), and producing a vasodilator with anti apoptotic and anti-inflammatory properties (CO). The reaction products of HO-1 may also have antifibrogenic properties. The purpose of this study is to explore the effects of HO-1 expression and its reaction products on fibronectin, an ECM protein, in the kidney. The results demonstrate that kidneys of HO-1 knockout mice express significantly more fibronectin protein as compared to heterozygote mice. A potent inducer of HO-1, hemin, significantly decreases fibronectin protein with a concomitant increase in HO-1 protein. Cells expressing HO-1, via TGF-beta1 induction, have reduced fibronectin expression. Bilirubin, a product of the heme oxygenase reaction, attenuates TGF-beta1-mediated increases in fibronectin expression. These results indicate that HO-1 induction and activity may modulate the production of ECM components and suggest a potential role for TGF-beta-mediated HO-1 induction in attenuating renal fibrosis. PMID- 16309586 TI - Heme oxygenase-1 transduction in endothelial cells causes downregulation of monocyte chemoattractant protein-1 and of genes involved in inflammation and growth. AB - Heme oxygenase (HO-1) has been implicated as an anti-inflammatory gene. HO-1 overexpression, transiently and chronically, affects heme protein expression, attenuates TNF-mediated cell death, and decreases adhesion molecules. We assessed the effect of oxidant-mediated agents such as glucose and heme on 8-epi isoprostane PGF2alpha (8-epi-PGF2alpha) and monocyte chemoattractant protein-1 (MCP-1). Glucose and heme increased both 8-epi-PGF2alpha and MCP-1. Overexpression of HO-1 decreased both 8-epi-PGF2alpha and MCP-1. To identify target genes involved in HO-1-mediated regulation of inflammation, a serial analysis of gene expression mRNA profile was performed in endothelial cells (EC) overexpressing the human HO-1 gene by transduction of a retrovirus carrying the HO-1 gene. Gene arrays (differential displays among 2400 genes) were used to identify known and novel differentially expressed genes. The levels of expression for several genes were confirmed by real time PCR in cells overexpressing the HO 1 gene. In HO-1 overexpressing cells, VEGF and the prostaglandin transporter were greatly increased while MCP-1 levels were decreased by 2.5-fold. The data from this study are relevant to understanding the mechanisms underlying the pathophysiological effects of HO-1 deficiency on endothelial cell injury and inflammation. PMID- 16309587 TI - Heme oxygenase-1 gene expression increases vascular relaxation and decreases inducible nitric oxide synthase in diabetic rats. AB - Hyperglycemia represents the main cause of complication of diabetes mellitus and oxidative stress, resulting from increased generation of reactive oxygen species (ROS), and plays a crucial role in their pathogenesis. Impairment of vascular responses in diabetic rats, as a result of an increase in superoxide (O2-), formation is a major complication in diabetes. Since heme oxygenase (HO) expression regulates the level of ROS by increasing antioxidant, such as glutathione and bilirubin, we investigated whether upregulation of HO-1 modulates the levels of iNOS and eNOS and altered vascular responses to phenylephrine (PE) and acetylcholine (Ach) in aorta and femoral arteries of diabetic (streptozotocin (STZ)-induced) rats. Our results showed that iNOS expression was increased, but HO activity was reduced, in diabetic compared to nondiabetic rats (p<0.05). Upregulation of HO-1 expression by cobalt protoporphyrin (CoPP), an inducer of HO 1 protein and activity, conferred an increase in eNOS and differentially decreased iNOS protein levels (p<0.05). Isolated aortic and femoral arteries obtained from diabetic rats exhibited contraction to PE and relaxation to Ach, which were markedly increased and decreased, respectively. However, HO-1 induction in diabetic rats normalized relaxation compared to controls. Therefore, overexpression of HO-1 may mediate an increase in eNOS and a decrease in iNOS, potentially contributing to restoration of vascular responses in diabetic rats. PMID- 16309588 TI - Oxidation of hemoglobin by lipid hydroperoxide associated with low-density lipoprotein (LDL) and increased cytotoxic effect by LDL oxidation in heme oxygenase-1 (HO-1) deficiency. AB - Heme-catalyzed oxidation of low-density lipoprotein (LDL) is one of the relevant mechanisms involved in LDL modification. We previously revealed a substantial oxidation of plasma hemoglobin to methemoglobin and a subsequent heme-catalyzed LDL oxidation generating moieties toxic to endothelium in heme oxygenase-1 (HO-1) deficiency in human. Drawing upon our previous observation we posited a pathway for oxidation of plasma hemoglobin in the HO-1-deficient child involving LDL associated lipid hydroperoxide. In support, LDL-associated lipid hydroperoxide oxidized ferrohemoglobin to methemoglobin--known to readily release its heme moieties--in a dose-dependent manner. Repeated heme exposure of the child s LDL further increased its lipid hydroperoxide content within min leading to additional cytotoxic effect on endothelium. Both cytotoxicity and HO-1 inducing ability of the oxidized LDL were strongly dependent on its lipid hydroperoxide content. We wondered if cells of the HO-1-deficient patient were prone to oxidative damage arising from heme-mediated oxidation of LDL. Indeed, we found elevated cytotoxicity induced by heme-catalyzed oxidation of LDL in lymphoblastoid cells derived from the HO-1-deficient patient. We conclude that oxidation of hemoglobin to methemoglobin by LDL-associated lipid hydroperoxide and increased sensitivity of cells of the HO-1-deficient child to stress of oxidized LDL might contribute to the vascular disorders reported earlier. PMID- 16309589 TI - Decreased levels of advanced glycation end-products in patients with Gilbert syndrome. AB - Oxidative stress plays an important role in the atherogenesis and bilirubin is one of the most potent antioxidant substances in human body. The aim of the present study was to investigate the impact of hyperbilirubinemia on plasma levels of advanced glycation end-products (AGEs), which contribute to atherogenesis. Two AGEs, pentosidine and Nepsilon-carboxymethyl lysine (CML), were determined in 23 subjects with Gilbert syndrome (GS) and in 21 age-matched healthy controls. Pentosidine was assessed by HPLC and CML was determined with ELISA. Logistic regression analysis was used for multiple adjustments of possible modifying factors. As expected, significantly higher serum bilirubin levels were found in GS subjects as compared to controls (28.9 +/- 9.6 vs 9.7 +/- 2.7 micromol/l, p<0.001). In contrast, serum levels of both AGEs were significantly lower in GS compared to normobilirubinemic controls (median; 25%-75% interquartile range (pentosidine: 1.12; 0.90-1.28 vs 1.31; 1.18-1.58 nmol/g protein, p<0.005; and CML: 6.70; 6.10-7.34 vs 7.33; 6.76-8.20 micromol/g protein, p = 0.01, respectively). Levels of both AGEs remained substantially lower even after adjustment for selected vascular risk and other modifying factors. In subjects with GS elevated serum bilirubin concentrations are associated with lower levels of AGEs. These results are in support of previous data on antioxidant properties of bilirubin. PMID- 16309590 TI - Protection from cardiac injury by induction of heme oxygenase-1 and nitric oxide synthase in a focal ischaemia-reperfusion model. AB - The enzymes heme oxygenase (HO) generate carbon monoxide (CO) in living organisms during heme degradation. Carbon monoxide has recently been shown to dilate blood vessels and to possess anti-inflammatory properties. It is also known that nitric oxide (NO) donors ameliorate cardiac ischaemia-reperfusion injury in experimental models of global or focal ischaemia-reperfusion (FIR). The two gaseous mediators share the same mechanism of action via the stimulation of soluble guanylyl cyclase and the increase in cellular levels of cyclic GMP. We studied the effects of manipulating the HO system and the possible interaction between CO and NO in an experimental in vivo model of FIR in the rat heart. FIR-subjected rats had necrotic area in the left ventricle, ventricular arrhythmias and a shortening of survival time in comparison to sham-operated animals. Resident mast cells underwent a heavy degranulation, malonyldialdehyde was produced by myocardial cell membranes, and tissue calcium levels were increased. High levels of myeloperoxidase were also detected, suggesting a FIR-related inflammatory process. In animals pre-treated with the HO-1 inducer, hemin, all the biochemical and morphometric markers of FIR were minimized or fully abated. Consistently, the biochemical and morphometric markers of FIR were reversed in rats treated with the HO-1 blocker, ZnPP-IX, prior to hemin administration. Pre-treatment with hemin significantly increases the expression and activity of both cardiac HO-1 and iNOS, suggesting that CO and NO cooperate in the cardioprotective effect against FIR-induced damage, and that there is a therapeutic synergism between NO donors and CO-releasing molecules, via the common stimulation of increase in cGMP levels and decrease in calcium overload. PMID- 16309591 TI - Carbon monoxide reduces the expression and activity of matrix metalloproteinases 1 and 2 in alveolar epithelial cells. AB - Matrix metalloproteinases (MMPs), particularly MMP-1 and MMP-2, are involved in the pathophysiology of emphysema. MMPs contain zinc in the catalytic site and its expression is regulated transcriptionally via mitogen activated protein kinases (MAPKs). Carbon monoxide (CO), one of the end products of heme oxygenase activity, has anti-inflammatory properties, which are mediated, at least in part, by activation of p38 MAPK. Furthermore, CO has the unique ability to bind to metal centers in proteins and can affect their specific activity. Therefore, we hypothesized that CO could inhibit MMPs expression and/or activity. Here we show that a recently identified carbon monoxide-releasing molecule, [Ru(CO)3Cl2]2 (or CORM-2) inhibits MMP-1 and MMP-2 mRNA expression in the human lung epithelial cell line A549. The MMPs mRNA expression was unaffected by the p38 MAPK inhibitor SB203580, but in the case of MMP-1 was reversed by the antioxidant N acetylcysteine. In addition, CORM-2 inhibited both MMP-1 and MMP-2 activities. Interestingly, no effect was observed with (Ru(DMSO)4Cl2), a negative control that does not contain CO groups. To the best of our knowledge this is the first evidence on the effect of CO on MMPs expression and activity. This effect could have important implications in the pathophysiology of emphysema and other diseases involving proteases/antiproteases imbalance. PMID- 16309592 TI - Reviewing the use of carbon monoxide-releasing molecules (CO-RMs) in biology: implications in endotoxin-mediated vascular dysfunction. AB - The inducible stress protein heme oxygenase-1 (HO-1) has been linked to tissue and organ protection against the deleterious actions of many pathological conditions, including endotoxin challenge. Similar protection can be achieved by the main products of heme oxygenase activity, namely bilirubin and carbon monoxide (CO). Since the identification of novel chemical compounds that liberate CO in biological systems (CO-releasing molecules or CO-RMs), our group and others have had access to a convenient and simple pharmacological tool that enables to study the role of CO in physiological functions. This article will review the scientific literature published to date on CO-RMs, with emphasis on the in vitro, ex vivo and in vivo experimental models employed to determine the contribution of CO to cellular mechanisms. In addition, we will report on the effect of heme oxygenase-related substances, such as bilirubin, CORM-3 and hemin, in a model of endotoxin-induced hypotension. Among the three different approaches examined, CORM-3 proved the most effective agent in reducing the fall in blood pressure caused by endotoxin. Furthermore, heme oxygenase-related substances affected the endotoxin-stimulated induction and distribution of hepatic HO-1 and inducible nitric oxide synthase (iNOS). Thus, it emerges that CO-RMs could exert important biological actions in the context of endotoxic-mediated dysfunction. PMID- 16309593 TI - Carbon monoxide-releasing molecules (CO-RMs) modulate respiration in isolated mitochondria. AB - Emerging evidence reveals that heme oxygenase-1 (HO-1) and its product carbon monoxide (CO) can exert diverse biological and cytoprotective effects. Our group has recently identified a new class of compounds (CO-releasing molecules or CO RMs) that can carry and deliver CO to biological systems and can be used to examine the physiological properties of CO. Here, we evaluated the influence of endogenously-generated CO (via HO-1 induction by hemin) and CO liberated from exogenously supplied CO-RMs on mitochondrial function. Renal mitochondria were isolated either from rats with increased HO-1 or from untreated animals, the latter being exposed to different concentrations of CO-RMs (10-100 microM). We found that mitochondrial oxygen uptake was significantly reduced in kidneys after HO-1 induction and, in a similar fashion, CO-RMs inhibited mitochondrial function in a concentration-dependent manner. Specifically, a marked depression of state 3 was observed resulting in a significant decrease in respiratory control index (RCI) values. When mitochondria were incubated with the inactive forms of CO-RMs, which are devoid of CO, the respiratory parameters remained unchanged. In summary, the results indicate that HO-1 induction and enhanced CO decrease renal oxygen consumption and alter mitochondrial function suggesting that CO could be a physiological regulator of mitochondrial oxidative phosphorylation. PMID- 16309594 TI - [Process approach experience of university trust]. AB - This job has the scope to make to comprise the importance of the quality system applied to a complex organization describing the phases that have characterized the definition and the improvement of one organizational-managerial procedure for the prevention and the treatment of a problem of great importance in the within of the care process (ulcer pressure), demonstrating as the personnel involvement is one of change strategy used . PMID- 16309595 TI - [The Role Playing with recording for the undoing of the relational abilities]. AB - The elective didactic activities, (the student choice) in the regulation nursing degree corse represent an inviting formative moment for the students, since they can choose on the didactic fan proposed, the offer that most like and capture one own learning process. The proposal of the Role Playing session, active didactic methodology for excellence, since it favours the learning taking advantage of ludic analogy power, has consented, in the elective activities a ambit, with a lot of favour by nurse students that have lived in the simulation some emblematic situations of the clinical daily. The students group has focused theme attention on some ordinary situations in the hospital context performed by actorsstudents and they are analysed psychological mechanism, particularly as far as concerned the communication and the approach nurse-patient-patient relative: the behaviour mistakes perceived by oneself and somebody else, are become elements for discussions and new information for to acquire relationship competences. PMID- 16309596 TI - [Mass-Media and nurses: opportunities and remembrances of an evolving professional]. AB - The actual Model that is developing in the relationship between who is giving (provider) and who is receiving health care, it is near to exceed the "modello paternalista" that for a long time used the citizen as people observing and collaborating, with the one were taking the decision according to the science. This kind of relationship, strongly without balancing, often produced an education of Health care operators, do not able to listen the people needs (Rotary International 2002) To defend this system someone stress the follow points: the patient does not have the right knowledge; the physical and mental conditions of the patient could be influenced, the different aims between patient and health care staff; the patient difficulty, to describe own feeling. The upper sentences shown an auto-referring System, do not well adapted to receive consent, to share aims, to satisfy needs; according to the respect of the sick (or patient), his values and his autonomy chosen; that comes from the developing of the social relationship. The communication is a tool that we can use to exceed this problem, if we understand the communication as a sharing and informative instrument the study leaded, shown how the nurses used the communication and its channels to share with the citizens the aims concerning the health; goals that the nursing acquired according to the principles declared in the Nurse/citizen Act. PMID- 16309597 TI - [Opinions and expectations of customers: an overview]. AB - The present article illustrates results of a survey, introduced in occasion of the celebration of the International Day of the Nurse, in the "Ospedale Niguarda Ca' Granda" of Milan, finalized to know the opinions and the expectations that the customers have developed in the comparisons of the nurses, taking into consideration Services and Operating Units. From it turns out that the customers have a much positive image of the nurse as a person who helps more than to be a professional. However when this dimension comes expressed from the customers, it very turns out defined and specified, independent regarding the medical profession. PMID- 16309598 TI - [Risk Management and analysis of an adverse event: accidental patients falls]. AB - The matter of safety of operators and users within health care is becoming more and more topical. The relationship between users and operators with the health structures is a very relevant and expensive circumstance. For this reason some years ago a new culture for clinic risk was born, called risk management (RM), its goal is to create prevention, which focuses on patients and on the risks they take at the moment in which they keep in contact with health structures. We have take into consideration the matter of accidental falls of hospitalized patients occurred in year 2003 at the Hospital of S. Croce of Fano (PU) and made a careful check of the report file prepared by the Health Management of the body mentioned. The unfavorable event we considered is resulted to be of modest entity (0,56% of falls on the total amount of patients hospitalized) uniformly distributed all year long. The most involved patients were male, almost all hospitalized in General Medicine and Geriatrics ward. Most of all individuals fallen down did not produce particular effects. The falls happened commonly during night. Patients who fell down are completely or partially self-supporting. Through a study of all data we collected we defined two variants which interact: on one side the quality of medical assistance, the nurse assistance in particular, and the characteristics of patients fallen down on the other side. Literature too confirm our statements and underline the need to identify the danger points to define the subjects in risk of fall. We found relevant under the present health background to focus our attention on health risks, in particular the accidental fall in hospitals, which has to be considered an important event carry out the monitoring the quality of health assistance. PMID- 16309601 TI - An analysis of iatrogenic complications from the total contact cast. AB - BACKGROUND: Many physicians continue to fear iatrogenic complications of the total contact cast, and use of this modality has remained restricted to a segment of the orthopaedic and podiatric communities. I examined the actual rate of complications and the factors associated with them in a large consecutive series of total contact casts drawn from the practice of a single surgeon to determine whether this modality is safe for multiple conditions in the neuropathic foot and ankle. METHOD: A consecutive series of 398 total contact casts spanning a 28 month period from the practice of a single physician were analyzed. All casts were placed by the same team of orthotists. This study comprised 70 patients with severe peripheral neuropathy; an average of 5.69 sequential casts per patient were placed. Three-hundred and sixty-seven casts were placed in diabetic patients, and 31 in patients with idiopathic peripheral neuropathy. By protocol, the initial cast was changed at an interval of no more than 1 week. On occasion, subsequent casts were left on longer, but on average casts were left in place for 7.69 days. Logistic regression analysis was used to analyze the contribution of patient factors to the chance of ulceration. RESULTS: Complications occurred in 22 casts, including six new pretibial ulcers, six new midfoot ulcers, four forefoot or toe ulcers, five hindfoot ulcers, and one malleolar ulcer. In no case was a preexisting ulcer made worse. This corresponds to an overall complication rate of 5.52% per cast. Overall, 30% of patients suffered one complication during the course of their treatment. With one exception, all new ulcers healed with simple modalities within 3 weeks, often with continued total contact casting. A single cast led to a proximal interphalangeal ulceration that exposed the joint and eventually required toe amputation. The rate of permanent sequelae from cast related injuries was therefore 0.25%. In no case were the resultant odds ratios statistically different from 1.0, but several trends were observed. Charcot arthropathy represented the highest risk (odds ratio 1.46), while the presence of neuropathic ulceration was surprisingly benign (odds ratio 0.69). The presence of diabetes as opposed to other causes of neuropathy was associated with increased risk (odds ratio 1.34). The use of a cast after deformity-correcting surgery in a neuropathic patient was remarkably safe (odds ratio 0.44), as were casts in which the patient was instructed to remain nonweightbearing (odds ratio 0.27). Patient age was not a factor (odds ratio 1.03). The length of time the cast was left on was not statistically important (odds ratio 0.99), although strict protocols for cast changing likely altered this data. CONCLUSION: A frequently changed total contact cast is a safe modality for the offloading and immobilization of the neuropathic foot, albeit with an expected constant rate of minor, reversible complications. Patients should be informed of these complications and risks before cast application. PMID- 16309602 TI - Arthroscopic treatment of anterior ankle impingement syndrome in dancers. AB - BACKGROUND: The purpose of this study was to evaluate the results of arthroscopic treatment of anterior bony and soft-tissue impingement of the ankle in elite dancers. METHODS: The study is a case series retrospectively reviewed. In the period between 1990 and 1999, 11 elite dancers (12 ankles) had ankle arthroscopy after a diagnosis of anterior ankle impingement that markedly interfered with their dancing. Initial nonoperative treatment failed in all subjects. Previous ankle trauma was noted in all subjects. There were seven women and four men (average age 28 years). Tibiotalar exostoses were radiographically noted in six ankles. Standard anteromedial and anterolateral arthroscopic portals and instrumentation were used for resection of bone spurs and debridement of impinging soft tissues. Patients were nonweightbearing for 5 days after surgery and had postoperative physiotherapy. RESULTS: Nine dancers returned to full dance activity at an average of 7 weeks after surgery. One patient did not return to dance performance because of concurrent unrelated orthopaedic problems, but he resumed work as a dance teacher; he developed a recurrent anterior tibial spur that was successfully resected at a second arthroscopy 9 years later. Another dancer developed postoperative scar-tissue impingement and stiffness; she had a repeat arthroscopy 4 months after the initial procedure and subsequently returned to dance performance. All patients eventually had marked postoperative improvement in pain relief and dance performance. CONCLUSIONS: Arthroscopic debridement is an effective method for the treatment of bony and soft-tissue anterior ankle impingement syndrome in dancers and has minimal morbidity. PMID- 16309603 TI - The modified Lapidus procedure for hallux valgus: a clinical and radiographic analysis. AB - BACKGROUND: Historically, the modified Lapidus procedure has been considered technically challenging, with high rates of complications, including nonunion and malunion. The purpose of this study was to review the clinical and radiographic results of this technique for the treatment of hallux valgus associated with first ray hypermobility, specifically examining patient satisfaction and the incidence of complications. METHODS: We retrospectively reviewed the results of the modified Lapidus procedure in 32 patients (38 feet). Evaluation included preoperative and postoperative questionnaires, physical examination, and radiographs. RESULTS: Complete clinical data was available for 29 patients (35 feet) and complete radiographic data for 29 patients (34 feet). Average age at surgery was 54 (range 27 to 84) years. Average followup was 42 months (range 29 to 93) months. Average preoperative visual analog pain score was 7.2 and postoperative 2.3 (p < 0.001). Average preoperative AOFAS Hallux MTP-IP Score was 44.8 and postoperative 87.3 (p < 0.001). Average preoperative intermetatarsal (IM) angle was 16 degrees, and the hallux valgus (HV) angle was 34 degrees. Postoperatively, the average IM angle was 6 degrees, the HV angle 11 degrees. There were no cases of nonunion or malunion. Complications included symptomatic hallux varus in two, recurrence of hallux valgus deformity in one, deep venous thrombosis in one, and failure of fixation in one patient. Twenty-four percent of patients (7 of 29) noted the subjective sensation of midfoot stiffness and 34% (10 of 29) noted forefoot stiffness. None of these patients thought that the stiffness was a disability. Ninety percent of patients (26 of 29) were satisfied with their foot function, and 86% (25 of 29) were satisfied with the cosmetic appearance of their foot. CONCLUSIONS: The modified Lapidus procedure results in a satisfactory clinical outcome in most patients. With meticulous operative technique, rigid internal fixation, and strict postoperative weightbearing restrictions, successful union can be achieved and complications can be minimized. Care should be taken to avoid hallux varus, and patients need to be counseled regarding a potentially long convalescent period and possible postoperative stiffness. PMID- 16309604 TI - Giannini prosthesis for flatfoot. AB - BACKGROUND: To determine the clinical and radiographic results of arthroereisis using the Giannini endo-orthotic implant for the treatment of pediatric flatfoot deformity, we retrospectively evaluated 37 patients (65 feet). METHODS: Twenty two boys and 15 girls were followed for an average of 26.5 months. Their overall average age was 9.4 (range 5 to 14) years. Pain, function, participation in sports activity, and the changes in radiographic measurements taken at 3, 6, and 12 months and then at 1-year intervals were evaluated. RESULTS: Pain or discomfort decreased from 60% (22 patients) preoperatively to 6% (2 patients) postoperatively. The percentage of normal postoperative footprints was 59% (38 feet) with first-degree flatfoot present in 27 feet (41%). Sports activities were taken up by 19 patients (51%) after surgery. An 8-mm endo-orthotic implant was used in 43 (66%) feet and an Achilles tenotomy was done in 38 feet (59%). The radiographic angles with the greatest degree of correction when compared to preoperative angles were the talar-first metatarsal angle (99%) and the calcaneal pitch (36%). After placement of the endo-orthotic implant, the talar angle influenced the rest of the radiographic measurements. There was no postoperative deterioration in any of the radiographic angles measured during the monitoring period. Complications occurred in 10.7% of the patients, with postoperative pain being the most frequently reported (6% of patients). There was no infection or local reaction to a foreign body. Removal of the endo-orthotic implant was not done on a routine basis. CONCLUSION: This operative technique respects the anatomical structure of the foot and produces good clinical and radiographic results. PMID- 16309605 TI - Cutaneous thermal sensitivity in diabetic neuropathy. AB - BACKGROUND: The goal of this investigation was to determine if cutaneous thermal sensitivity could be used as a discriminator of peripheral neuropathy in diabetic subjects who were sensate to the Semmes-Weinstein 5.07 monofilament. METHODS: Sixty adult subjects were separated into two groups. The control group (A) was composed of 30 young healthy individuals without a history of diabetes. The focus group (B) was composed of 26 individuals with adult onset diabetes and four with juvenile onset. All of the subjects underwent thermal sensitivity testing in peripheral nerve root dermatomes of their hands and feet. Testing was performed with custom devices fabricated from materials with different thermal conduction capacities (copper, steel, glass, and plastic). Similar tests were performed with glass tubes containing heated or cooled water to develop a range of thermal sensitivity for the subjects. RESULTS: There was a strong relationship between cold perception and stimulation with the copper probe in dermatomes of the radial nerve of the upper limb and the superficial peroneal dermatome of the lower limb. CONCLUSIONS: Thermal sensitivity to copper and cold stimulation may be more discriminative and have a higher threshold than sensitivity to the Semmes Weinstein monofilament. This simple method may have a role in the early detection of peripheral neuropathy in adult-onset diabetes mellitus. PMID- 16309606 TI - The incidence of Achilles tendon ruptures in Edmonton, Canada. AB - BACKGROUND: The incidence of Achilles tendon ruptures specific to the North American population has not been previously reported and current epidemiological data are primarily reported from European communities. The purpose of this study was to determine the incidence of Achilles tendon ruptures in the city of Edmonton, Alberta, Canada, and to compare this data to those reported in European studies. METHODS: A retrospective chart review from all five acute care hospitals in Edmonton from 1998 to 2002 (inclusive) were reviewed for Achilles tendon ruptures. Data such as gender, age, side, mechanism of injury, and season of injury were obtained. RESULTS: The incidence of Achilles tendon ruptures ranged from an annual average of 5.5 ruptures to 9.9 ruptures per 100,000 inhabitants with an overall mean of 8.3 ruptures per 100,000 people. There was a statistically significant difference in Achilles tendon ruptures over the last two study years for both genders (women, p < 0.02; men, p < 0.03). The mean age for an Achilles tendon rupture was 40.6 years for men and 44.5 years for women. The Achilles tendon ruptures occurred most frequently in the 30 to 39 and 40 to 49 year old age groups in both men and women, respectively (p < 0.02). Most ruptures occurred in the spring season, but there was no statistical difference in the incidence of Achilles tendon ruptures by season (p > 0.05). CONCLUSIONS: The incidence of Achilles tendon ruptures in this community was comparable to those reported in European communities (range 6 to 37 ruptures per 100,000 people), although a bimodal age distribution of rupture previously reported was not observed in this study. PMID- 16309607 TI - The radiographic morphology of foot deformities in patients with fibrodysplasia ossificans progressiva. AB - BACKGROUND: The most consistent deformities that allow early diagnosis of fibrodysplasia ossificans progressiva are the presence of bilateral short first rays and hallux valgus. The purpose of this study was to describe the radiographic features observed in the feet of patients with fibrodysplasia ossificans progressiva. METHODS: The radiographs of 26 feet (15 patients with fibrodysplasia ossificans progressiva) were reviewed to evaluate the radiographic changes that occur in the first ray. Variables analyzed were the hallux valgus (HV) angle, the distal metatarsal articular (DMA) angle, the proximal phalangeal articular (PPA) angle, the intermetatarsal (IM) angle, ratio of the lengths of the first and second metatarsal lengths (MT1:MT2), and the first and second ray length ratio. The length ratios were then subtracted from similar ratios in radiographs of age- and gender-matched normal patients previously reported. RESULTS: The proximal phalanx was consistently shortened but morphologically dissimilar from subject to subject. Asymmetry was noted in some patients with bilateral radiographs. The mean HV angle was 28 degrees, and the mean IM angle was 10 degrees. The mean DMA angle was 33 degrees, and the mean PPA angle was 14 degrees. The MT1:MT2 ratio was 0.89, and the mean first ray to second ray length ratio was 0.87. The mean of the differences in the MT1:MT2 and first and second ray length ratios in patients with fibrodysplasia ossificans progressiva compared to the normal controls were 0.05 and 0.01, respectively. Fusion occurred between the abnormal tibial epiphysis of the proximal phalanx and metatarsal head with advancing age, and 68% of the metatarsal heads were fused with the abnormal proximal phalangeal epiphysis. CONCLUSIONS: Foot pathology in patients with fibrodysplasia ossificans progressiva is variable but consistently involves an abnormality of the tibial aspect of the proximal phalangeal epiphysis of the hallux. This results in the clinical observation of hallux valgus in these patients. The first metatarsal is consistently shortened, and fusion between the epiphysis of the abnormal proximal phalanx and the shortened first metatarsal head occurs with advancing age. PMID- 16309608 TI - The variations of the sensory branches of the superficial peroneal nerve course and its clinical importance. AB - BACKGROUND: Although the sensory branches of the superficial peroneal nerve (SPN) have different anatomical variations that are of clinical importance, little is known about their anatomic courses, branching patterns, or relationships to palpable osseous landmarks.(1,3) A detailed knowledge is necessary for surgical exposures about the foot and ankle, arthroscopic procedures, ankle block anesthesia, and SPN block for leg venography. METHODS: Thirty lower cadaver limbs were dissected to assess the anatomic properties and the variations of the sensory branches of the SPN. RESULTS: Three distinct branch patterns were determined. In Type 1 (63.3%), the nerve penetrated the crural fascia 80.15 +/- 17.80 mm proximal to the intermalleolar line and then divided into the intermediate dorsal cutaneous nerve (IDCN) and the medial dorsal cutaneous nerve (MDCN) (classic type). In Type 2 (26.7%), the IDCN and MDCN arose independently from the SPN. In Type 3 (10%), the SPN penetrated the crural fascia 101.14 +/- 70.27 mm proximal to the intermalleolar line as a single branch. This single branch had a similar course to the MDCN. Measurements in this study were obtained from palpable bony reference landmarks. CONCLUSION: Detailed knowledge about the SPN, IDCN, and the MDCN may decrease the damage to these nerves during operative procedures near the foot and ankle. PMID- 16309609 TI - The peroneal tubercle: description, classification, and relevance to peroneus longus tendon pathology. AB - BACKGROUND: The size and configuration of the peroneal tubercle has been implicated in the pathogenesis of peroneal tendon tears and tenosynovitis. The purpose of this study was to determine the size and prevalence of the peroneal tubercle and devise a classification scheme according to the structure of the tubercle. METHODS: One hundred and seventeen calcanei were selected from 59 human skeletons in an osteological collection (one calcaneus was missing). Three were excluded because of the poor condition, leaving 114 calcanei. The peroneal tubercle was measured in length, height, and depth and its structure subjectively described as flat, prominent, concave, or tunnel for each specimen when present. RESULTS: This study revealed a peroneal tubercle prevalence of 90.4% (103) in 114 calcanei. The average length, height, and depth of the tubercle were 13.04 mm (range 3.61 mm to 26.66 mm), 9.44 mm (range 3.67 mm to 23.40 mm), and 3.13 mm (range 1 to 10), respectively. The peroneal tubercle was classified structurally as flat in 44 (42.7%), prominent in 30 (29.1%), concave in 28 (27.2%), and tunnel in one (1.0%). CONCLUSION: This data may further help to understand the size and assorted configurations of the peroneal tubercle and how they relate to peroneus longus tendon pathology. PMID- 16309610 TI - The validity and reliability of the Klaue device. AB - BACKGROUND: Excessive first ray mobility has been implicated as the cause of many forefoot abnormalities. The association between hypermobility and forefoot pathology is controversial, and this is largely related to the difficulty in quantifying first ray motion. Manual examinations have been shown to be unreliable. Klaue etal. developed a device consisting of a modified ankle-foot orthosis with an attached micrometer to objectively measure first ray mobility. The purpose of this study was to evaluate the validity and reliability of this device. METHODS: Sixteen fresh-frozen, below-knee amputation specimens with hallux valgus were used for the study. The study was divided into two parts. Part I was an analysis of the validity of the Klaue device; first ray dorsal displacement was measured on lateral radiographs following manual manipulation, and values were statistically compared to the Klaue device measurements. Part II of the study was an evaluation of intraobserver and interobserver agreement. Two clinicians used the Klaue device on each of the cadaver limbs, and values of first ray sagittal mobility were recorded and compared. RESULTS: The mean value of first ray mobility measured with the Klaue device was 7.5 mm and the average displacement measured from the lateral radiographs was 7.4 mm. Paired t-testing showed no significant difference between the Klaue and radiographic measurements (p = 0.83). The mean first ray mobility by examiners 1 and 2 with the Klaue device were identical (10.5 mm), and statistical analysis showed no significant interobserver or intraobserver differences. CONCLUSIONS: The results confirm the validity of the Klaue device and limited variability of measurements between experienced users. PMID- 16309611 TI - A comparison of device measures of dorsal first ray mobility. AB - BACKGROUND: Devices built by Glasoe and Klaue have been used in several studies to measure first ray mobility. Both devices measure sagittal motion of the first ray in a dorsal direction. The primary difference in the devices is the method of the load imposed. This study investigates whether first ray mobility measured with the Glasoe device is similar to the amount of mobility measured with the Klaue device. METHODS: Using the devices described by Glasoe and Klaue, dorsal first ray mobility was measured in 39 patients who had foot and ankle problems. Paired t-tests were computed to assess for differences between device measures of dorsal mobility. Intraclass correlation coefficient (ICC) and absolute difference values were computed to further assess the agreement in measures. RESULTS: Dorsal mobility measured with the Glasoe device averaged 4.9 mm (1.8 to 9.3 mm). Dorsal mobility measured with the Klaue device averaged 5.2 mm (2.5 to 8.5 mm). Paired t tests (p = 0.12) revealed no significant difference in measures. An ICC of 0.70 and a mean absolute difference of 0.9 mm (SD 0.8) were found between the two clinical measures further suggesting agreement. CONCLUSION: Results indicated that the two devices possess similar diagnostic accuracy in the measurement of dorsal first ray mobility. PMID- 16309612 TI - Reliability of the Foot Function Index:: A report of the AOFAS Outcomes Committee.. AB - BACKGROUND: There currently is no widely used, validated, self-administered instrument for measuring musculoskeletal functional status in individuals with nonsystemic foot disorders. The purpose of this paper was to report on the assessment of reliability of one of these instruments. We wanted to determine if the Foot Function Index (FFI), which has been validated in rheumatoid patients without fixed foot deformity or prior foot surgery, would be reliable for a population of patients with foot complaints without systemic disease. METHODS: Patients were recruited from five orthopaedic offices where the physicians were members of the American Orthopaedic Foot and Ankle Society. Patients were asked to complete the FFI at the time of their initial office visit and then were given a second copy to complete and return by mail 1 week after their visit. RESULTS: Ninety-six patients completed the first questionnaire, and 54 patients completed the second. Reliability in this population was acceptable with an average of 23.5% of the patients providing retest values within one point of the initial response and an average of 45.3% of the patients providing the same response, for a total of 68.8% of all respondents answering within one point between their initial and second questionnaire. In two of the three categories, there were frequent nonresponses or no applicable responses. Four questions, two in the pain section and two in the activity limitation section, generated 20% or more of the nonapplicable answers. CONCLUSIONS: The FFI appears to be a reasonable tool for low functioning individuals with foot disorders. It may not be appropriate for individuals who function at or above the level of independent activities of daily living. PMID- 16309613 TI - Evidence of validity for the Foot and Ankle Ability Measure (FAAM). AB - BACKGROUND: There is no universally accepted instrument that can be used to evaluate changes in self-reported physical function for individuals with leg, ankle, and foot musculoskeletal disorders. The objective of this study was to develop an instrument to meet this need: the Foot and Ankle Ability Measure (FAAM). Additionally, this study was designed to provide validity evidence for interpretation of FAAM scores. METHODS: Final item reduction was completed using item response theory with 1027 subjects. Validity evidence was provided by 164 subjects that were expected to change and 79 subjects that were expected to remain stable. These subjects were given the FAAM and SF-36 to complete on two occasions 4 weeks apart. RESULTS: The final version of the FAAM consists of the 21-item activities of daily living (ADL) and 8-item Sports subscales, which together produced information across the spectrum ability. Validity evidence was provided for test content, internal structure, score stability, and responsiveness. Test retest reliability was 0.89 and 0.87 for the ADL and Sports subscales, respectively. The minimal detectable change based on a 95% confidence interval was +/-5.7 and +/--12.3 points for the ADL and Sports subscales, respectively. Two-way repeated measures ANOVA and ROC analysis found both the ADL and Sports subscales were responsive to changes in status (p < 0.05). The minimal clinically important differences were 8 and 9 points for the ADL and Sports subscales, respectively. Guyatt responsive index and ROC analysis found the ADL subscale was more responsive than general measures of physical function while the Sports subscale was not. The ADL and Sport subscales demonstrated strong relationships with the SF-36 physical function subscale (r = 0.84, 0.78) and physical component summary score (r = 0.78, 0.80) and weak relationships with the SF-36 mental function subscale (r = 0.18, 0.11) and mental component summary score (r = 0.05, -0.02). CONCLUSIONS: The FAAM is a reliable, responsive, and valid measure of physical function for individuals with a broad range of musculoskeletal disorders of the lower leg, foot, and ankle. PMID- 16309614 TI - Screw fixation compared to H-locking plate fixation for first metatarsocuneiform arthrodesis: a biomechanical study. AB - BACKGROUND: Several different techniques have been used for fixation of first metatarsocuneiform (MTC) joint arthrodesis, a standard treatment for arthritis, instability, and deformity of the MTC joint. Improved plating systems using locking designs are now available, but no studies have yet compared this construct with other methods. We compared load to failure with a locking plate design versus standard crossed-screw fixation. METHODS: Ten matched pairs of fresh frozen cadaver feet were used. The bone density of each pair was measured with DEXA scanning. One foot of each pair was randomly assigned to have a dorsomedial Normed H titanium locking plate (Normed Medizin-Technik Vertriebs GmbH, D-78501 Tuttlingen, Germany) applied to the first MTC joint. On the other foot of the pair, fixation of the first MTC joint was done with crossed ACE DePuy 4.0 (DePuy/Ace, Warsaw, IN) titanium cannulated screws. The first metatarsal and first cuneiform were then isolated and planted in an epoxy resin. The specimens were loaded to failure in a four-point bending configuration using a MTS Mini Bionix test frame (MTS Systems Corp., Eden Prairie, MN). Failure was defined as displacement of more than 3 mm at the arthrodesis site. The Student t-test was used to determine any observed differences, with significance set at p CH2 eliminations at the cis-terminal methyl in all cases; (2) similar primary kinetic isotope effects (KIE) of 4.25-4.64 for the CH3-->CH2 eliminations; (3) a significant intermolecular KIE (1.33+/-0.03) in competitive cyclizations of unlabeled FPP and FPP-d6 to premnaspirodiene by HPS; (4) stereoselective incorporation of label from D2O into the 1beta position of epiaristolochene; (5) stereoselective eliminations of the 1beta and 9beta protons in formation of epiaristolochene and its delta(1(10)) isomer epieremophilene by TEAS and CH4; and (6) predominant loss of the 1alpha proton in forming the cyclohexene double bond of premnaspirodiene by HPS and CH4. The results are explained by consideration of the conformations of individual intermediates, and by imposing the requirement of stereoelectronically favorable proton additions and eliminations. PMID- 16309623 TI - Structure-activity relationship of an alpha-toxin Bs-Tx28 from scorpion (Buthus sindicus) venom suggests a new alpha-toxin subfamily. AB - Scorpion venoms are among the most widely known source of peptidyl neurotoxins used for callipering different ion channels, e.g., for Na(+), K(+), Ca(+) or Cl( ). An alpha-toxin (Bs-Tx28) has been purified from the venom of scorpion Buthus sindicus, a common yellow scorpion of Sindh, Pakistan. The primary structure of Bs-Tx28 was established using a combination of MALDI-TOF-MS, LC-ESI-MS, and automated Edman degradation analysis. Bs-Tx28 consists of 65 amino acid residues (7274.3+/-2Da), including eight cysteine residues, and shows very high sequence identity (82-94%) with other long-chain alpha-neurotoxins, active against receptor site-3 of mammalian (e.g., Lqq-IV and Lqh-IV from scorpions Leiurus sp.) and insect (e.g., BJalpha-IT and Od-1 from Buthotus judaicus and Odonthobuthus doriae, respectively) voltage-gated Na(+) channels. Multiple sequence alignment and phylogenetic analysis of Bs-Tx28 with other known alpha- and alpha-like toxins suggests the presence of a new and separate subfamily of scorpion alpha toxins. Bs-Tx28 which is weakly active in both, mammals and insects (LD(50) 0.088 and 14.3microg/g, respectively), shows strong induction of the rat afferent nerve discharge in a dose-dependent fashion (EC(50)=0.01microg/mL) which was completely abolished in the presence of tetrodotoxin suggesting the binding of Bs-Tx28 to the TTX-sensitive Na(+)-channel. Three-dimensional structural features of Bs Tx28, established by homology modeling, were compared with other known classical alpha-mammal (AaH-II), alpha-insect (Lqh-alphaIT), and alpha-like (BmK-M4) toxins and revealed subtle variations in the Nt-, Core-, and RT-CT-domains (functional domains) which constitute a "necklace-like" structure differing significantly in all alpha-toxin subfamilies. On the other hand, a high level of conservation has been observed in the conserved hydrophobic surface with the only substitution of W43 (Y43/42) and an additional hydrophobic character at position F40 (L40/A/V/G39), as compared to the other mentioned alpha-toxins. Despite major differences within the primary structure and activities of Bs-Tx28, it shares a common structural and functional motif (e.g., transRT-farCT) within the RT-CT domain which is characteristic of scorpion alpha-mammal toxins. PMID- 16309624 TI - The effect of non-enzymatic glycation on the unfolding of human serum albumin. AB - We monitored the unfolding of human serum albumin (HSA) and glycated human serum albumin (gHSA) subjected to guanidine hydrochloride (GndHCl) by using fluorescence and circular dichroism (CD) spectroscopy. A two-state model with sloping baselines best described the Trp-214 fluorescence unfolding measurements, while a three-state model best described the far-UV CD unfolding data. Glycation of HSA increased the [D](50%) point by approximately 0.20M. This corresponded to an increase in the free energy of unfolding of gHSA relative to HSA of 2.6kJ/mol. The intrinsic fluorescence of Trp-214 in gHSA is 0.72 of that of HSA and the far UV CD spectrum of gHSA is nearly identical to that of HSA. These results showed that glycation altered the local structure around Trp-214 while not significantly impacting the secondary structure, and this alteration translated into an overall change in the stability of gHSA compared to HSA. PMID- 16309625 TI - Elevated expression of alphaA- and alphaB-crystallins in streptozotocin-induced diabetic rat. AB - alpha-Crystallin, a predominant protein of the ocular lens, is composed of two subunits, alphaA and alphaB. Of these, alphaB-crystallin has been shown to present widely in non-lenticular tissues while alphaA-crystallin is largely lens specific. Although, expression of alphaB-crystallin is elevated under various stress and pathological conditions, yet its physiological significance remained unknown. Some studies suggest that the expression of alphaB-crystallin gene is related to oxidative stress. Persistent hyperglycemia during uncontrolled diabetes is known to cause oxidative stress, which has been implicated in various secondary complications of diabetes. Hence, expression of alphaA- and alphaB crystallins in various tissues of streptozotocin (STZ)-induced diabetic Wistar NIN rats was investigated by RT-PCR and immunoblotting. While expression of alphaB-crystallin was noted in the wide range of tissues examined in the study, alphaA-crystallin expression was detected only in lens and retina. Interestingly, alphaB-crystallin expression was elevated in lens, heart, muscle, and brain, but decreased in adipose tissue of diabetic rats compared to control rats. alphaA Crystallin expression was increased in retina of diabetic rat. Increased oxidative stress appears to be a major stimulus for the enhanced expression of alphaA- and alphaB-crystallins in the tissues of diabetic rats and elevated expression of alpha-crystallin may have a protective role against metabolic stress. Interestingly, feeding of curcumin, a dietary antioxidant, to diabetic rats attenuated the enhanced expression of alphaB-crystallin. The results indicate that elevated expression of alpha-crystallins in some tissues may have implications in pathophysiology of diabetic complications. PMID- 16309626 TI - Cold, salinity and drought stresses: an overview. AB - World population is increasing at an alarming rate and is expected to reach about six billion by the end of year 2050. On the other hand food productivity is decreasing due to the effect of various abiotic stresses; therefore minimizing these losses is a major area of concern for all nations to cope with the increasing food requirements. Cold, salinity and drought are among the major stresses, which adversely affect plants growth and productivity; hence it is important to develop stress tolerant crops. In general, low temperature mainly results in mechanical constraint, whereas salinity and drought exerts its malicious effect mainly by disrupting the ionic and osmotic equilibrium of the cell. It is now well known that the stress signal is first perceived at the membrane level by the receptors and then transduced in the cell to switch on the stress responsive genes for mediating stress tolerance. Understanding the mechanism of stress tolerance along with a plethora of genes involved in stress signaling network is important for crop improvement. Recently, some genes of calcium-signaling and nucleic acid pathways have been reported to be up-regulated in response to both cold and salinity stresses indicating the presence of cross talk between these pathways. In this review we have emphasized on various aspects of cold, salinity and drought stresses. Various factors pertaining to cold acclimation, promoter elements, and role of transcription factors in stress signaling pathway have been described. The role of calcium as an important signaling molecule in response to various stress signals has also been covered. In each of these stresses we have tried to address the issues, which significantly affect the gene expression in relation to plant physiology. PMID- 16309627 TI - Mechanism of interaction of PITPalpha with membranes: conformational changes in the C-terminus associated with membrane binding. AB - Eukaryotic phosphatidylinositol transfer proteins (PITPs) are composed predominantly of small ( approximately 32 kDa) soluble proteins that bind and transfer a single phospholipid, normally phosphatidylinositol or phosphatidycholine. Two forms, PITPalpha and PITPbeta, which share approximately 80% amino acid sequence similarity, are known. Rat PITPalpha was labeled at specific single reactive Cys residues with I-AEDANS and used to examine PITP membrane interactions. Upon binding to phospholipid vesicles, PITP labeled with AEDANS at the C-terminus, a region postulated to be involved in membrane binding, shows significant decreases in both steady-state and dynamic fluorescence anisotropy. In contrast, PITPs labeled with AEDANS at sites located distal to the C-terminus show increases in both steady-state and dynamic anisotropy. These results suggest that interaction of PITP with membrane surfaces leads to significant alterations in conformation and perhaps melting of the C-terminal helix. PMID- 16309628 TI - Enhancement of lactate metabolism in the basolateral amygdala by physical and psychological stress: role of benzodiazepine receptors. AB - Lactate is considered to play a significant role in energy metabolism and reflect neural activity in the brain. Using in vivo microdialysis technique, we measured extracellular lactate concentrations in the basolateral amygdaloid nucleus (BLA) of rats under electric footshock or psychological stress. We also attempted to determine whether the stress-induced changes of extracellular lactate concentrations in the BLA are attenuated by diazepam, an agonist at benzodiazepine receptors, and whether FG7142, an inverse agonist at benzodiazepine receptors, have a facilitative effect on energy metabolism in the BLA. Both footshock and psychological stress led to an increase in extracellular lactate concentrations in the BLA. Similar increment of extracellular lactate levels was observed by administration of FG7142. Pretreatment with diazepam attenuated the ability of FG7142, as well as physical or psychological burden, to increase lactate levels in the BLA. These results indicate that a variety of stressors enhances energy metabolism in the BLA, and suggest that some stress induced changes in energy metabolism are regulated by benzodiazepine receptors. PMID- 16309629 TI - Geranylgeranylacetone, a heat shock protein inducer, prevents acoustic injury in the guinea pig. AB - Geranylgeranylacetone (GGA) used widely as anti-ulcer agent is accepted as an inducer of the heat shock proteins (Hsps) at gastric mucosa, liver, heart, and brain. However, there have been no reports that GGA could induce Hsps in the cochlea leading up to the oto-protection. The purpose of the present study was to investigate whether single oral dose of GGA could induce Hsps at cochlea and oral administration had protective effect to the cochlea against noise trauma. We used Hartley guinea pigs and investigated the expression of Hsp70, 40, and 27 in cochlea by Western blot analysis. To evaluate cochlear function, we assessed thresholds of the auditory brain stem response (ABR). For histological assessment, we observed the sensory epithelium using surface preparation technique. GGA (600 mg/kg) or vehicle was given orally to animals. Western blot analysis showed that the expressions of Hsp 70, 40, and 27 were increased 24-48 h after administration of single dose of GGA, whereas there was less expression in the animals given vehicle. In the animals given GGA once a day for a week before sound exposure (130 dB SPL octave band noise with a center frequency of 4 kHz) for 3 h, their ABR threshold elevations were lowered significantly. In addition, significantly fewer defects were observed on outer hair cells of organ of Corti in the animals treated by GGA than those of the animals without GGA. This result shows that pretreatment by GGA have a potential to prevent cochlea damage against the intense noise. PMID- 16309630 TI - Sleep restriction by forced activity reduces hippocampal cell proliferation. AB - Mounting evidence suggests that sleep loss negatively affects learning and memory processes through disruption of hippocampal function. In the present study, we examined whether sleep loss alters the generation, differentiation, and survival of new cells in the dentate gyrus. Rats were sleep restricted by keeping them awake in slowly rotating drums for 1 day or repeatedly for 20 h/day over a period of 8 days. In addition to home cage controls, we included forced activity controls which, compared to sleep restricted rats, walked at double speed for half the time. These animals thus walked the same distance but had sufficient time to sleep. The results show that a single day of sleep deprivation significantly reduced hippocampal cell proliferation in the hilus of the dentate gyrus as measured by immunostaining for the proliferation marker Ki-67. Repeated partial sleep deprivation reduced cell proliferation in both the hilus and the subgranular zone. However, the latter was also found after chronic forced activity, and may not have been specific for sleep loss. To study neuronal survival, rats received a single intraperitoneal injection of 5-bromo-2' deoxyuridine (BrdU) 5 days before the experiment. The number of surviving, BrdU positive cells was not affected by sleep restriction. Also, the differentiation of BrdU-positive new cells into NeuN-positive neuronal and GFAP-positive glial phenotypes was not significantly altered by sleep loss. In conclusion, since new cells in the hilus mostly differentiate into glia, our findings indicate that sleep loss may reduce hippocampal gliogenesis. PMID- 16309631 TI - Time of day alters long-term sensitization to cocaine in rats. AB - To test the hypothesis that time of day influences the locomotor effects of cocaine in rats, we measured short- and long-term sensitization to cocaine at five different Zeitgeber times (ZT). Short-term sensitization was expressed equally at all ZTs tested. Long-term sensitization was expressed only at dark onset, suggesting a possible influence of melatonin. Exogenous melatonin given immediately before cocaine challenge slightly enhanced long-term sensitization. Taken together, the present results support a time-of-day influence on locomotor sensitization to cocaine. PMID- 16309632 TI - Regulation of activity-regulated cytoskeleton protein (Arc) mRNA after acute and chronic electroconvulsive stimulation in the rat. AB - The temporal profile of Arc gene expression after acute and chronic electroconvulsive stimulations (ECS) was studied using semi-quantitative in situ hybridisation in the rat cortex. A single ECS strongly and temporarily increased Arc mRNA levels in dentate granular cells with maximal induction seen up to 4 h after the stimulus, but returned to baseline at 24 h. A single ECS also increased expression of Arc mRNA in the CA1 and the parietal cortex, but the expression peaked within 1 h and returned to baseline levels within 2 h. Repeated or chronic ECS is a model of electroconvulsive therapy and it would be predicted that gene products involved in antidepressant effects accumulate after repeated ECS. However, repeated ECS reduced Arc gene expression in the CA1 24 h after the last stimulus. These results indicate that Arc is an immediate early gene product regulated by an acute excitatory stimulus, but not accumulated by long term repetitive ECS and therefore not a molecular biomarker for antidepressant properties. More likely, Arc is likely a molecular link to the decline in memory consolidation seen in depressive patients subjected to electroconvulsive therapy. PMID- 16309633 TI - Capsaicin evokes hypothermia independent of cannabinoid CB1 and CB2 receptors. AB - The present study investigated a potential role for cannabinoid CB(1) and CB(2) receptors in capsaicin-evoked hypothermia. Capsaicin (1 mg/kg, s.c.) caused rapid and significant hypothermia in rats. Pretreatment with SR 141716A (1, 2.5 and 5 mg/kg, i.p.), a CB(1) antagonist, or SR 144528 (1, 2.5 and 5 mg/kg, i.p.), a CB(2) antagonist, did not affect capsaicin-induced hypothermia. In separate experiments, the hypothermia caused by WIN 55212-2 (5 mg/kg, i.m.), a cannabinoid agonist, was not significantly altered by capsazepine (10 and 30 mg/kg, i.p.) or SB 366791 (2 mg/kg, i.p.), a novel TRPV1 antagonist. These data suggest that capsaicin causes hypothermia by a CB(1)- and CB(2)-independent mechanism, and that WIN 55212-2 causes hypothermia by a TRPV1-independent mechanism. PMID- 16309634 TI - The effect of P2X receptor activity on GABAA receptor-mediated inhibition in the gerbil hippocampus. AB - In the present study, to elucidate the effect of altered P(2)X receptor transmission on GABA(A) receptor expression and its transmission, we studied the morphological and electrophysiological responses of GABA(A) receptor in the gerbil hippocampus following P(2)X receptor antagonist/agonist treatment. Suramin or pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) treatment did not affect GABA(A) receptor immunoreactivities and paired-pulse responses in the gerbil hippocampus. In addition, ATP treatment did not significantly affect population spike amplitude ratios and EPSP slope ratios in the gerbil dentate gyrus. Co-application, but not pretreatment, of PPADS or suramin enhanced the effect of muscimol on paired-pulse inhibition in the dentate gyrus. In contrast, co-application of ATP reduced the effect of muscimol in the dentate gyrus. These findings indicate that the blockade of P(2)X receptor did not affect GABA(A) receptor immunoreactivities, and P(2)X receptor may modulate GABA(A) receptor mediated inhibition when in co-activation with GABA(A) receptor. Therefore, our findings suggest that the relationship between GABA(A) receptor and P(2)X receptor may not be reciprocal, although GABA(A) receptor activity affects P(2)X receptor functionality and its expression. PMID- 16309635 TI - Caspase-mediated cell death predominates following engraftment of neural progenitor cells into traumatically injured rat brain. AB - Neural progenitor cells (NPCs) have been shown to be a promising therapy for cell replacement and gene transfer in neurological diseases including traumatic brain injury (TBI). However, NPCs often survive poorly after transplantation despite immunosuppression, and the mechanisms of graft cell death are unknown. In this study, we evaluated caspase- and calpain-mediated mechanisms of cell death of neonatal mouse C17.2 progenitor cells, transplanted at 24 h following lateral fluid percussion brain injury (FP) in rats. Adult Male Sprague-Dawley rats (n = 30) were subjected to lateral FP injury (n = 18) or sham surgery (n = 12). C17.2 cells labeled with green fluorescent dye (CMFDA) were engrafted in the perilesional deep cortex, and animals were sacrificed at 24 h, 72 h and 1 week post-transplantation. Pro-apoptotic caspase-mediated cleavage products (Ab246) and calpain-mediated cleavage products (Ab38) were detected in the engrafted cells using immunohistochemistry. Only 2 to 4.5% of grafted NPCs were found to survive at 24 h post-transplantation, regardless of injury status of the host brain, although brain-injured animals had significantly fewer graft cells than sham-injured animals. Limited caspase and calpain-mediated graft cell death was observed in both sham- and brain-injured animals, and caspase-mediated graft cell death was significantly greater than calpain-mediated graft cell death in all animals. Brain-injured animals had significantly increased caspase-mediated graft cell death compared to sham-injured animals. These results suggest that both the caspase and calpain family of proteases are involved in graft cell death, and that caspase-mediated apoptotic graft cell death predominates in the acute post traumatic period following TBI. PMID- 16309636 TI - Differential regulation of cortical NMDA receptor subunits by sensory learning. AB - NMDA receptor is an important player in neuronal plasticity, including cortical reorganization. In the adult cerebral cortex, the receptor properties are regulated by relative expression of NR2A and NR2B subunits. We have previously found that 3 days of sensory conditioning, in which stimulation of whiskers was paired with a tail shock, induce NMDA-receptor-dependent expansion of metabolically labeled cortical representations of the stimulated vibrissae. Here, we examined the effect of learning-induced cortical reorganization upon expression of NR2A and NR2B NMDA receptor subunits. An increase in NR2A mRNA expression in the barrel of the "trained" row of vibrissae was observed with in situ hybridization 24 h after sensory conditioning. NR2B mRNA expression level did not change. Protein level of both regulatory subunits and obligatory NR1 subunit were examined in P2 fraction. NR2A protein level was found elevated 1 h and 24 h after the sensory conditioning, but not in controls which received only whisker stimulation, signifying that the change was associated with cortical map reorganization. NR2B protein level was transiently elevated in both trained and stimulated control groups. NR1 protein level did not change. The results show that simple sensory learning induces a change in expression of regulatory NMDA receptor subunits, indicating a potential for receptor channel properties modification. PMID- 16309663 TI - LDL susceptibility to oxidation and HDL antioxidant capacity in patients with renal failure. AB - OBJECTIVES: This study examines the susceptibility to oxidation and the ability to stimulate reactive oxygen species of LDL from hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients. It was also designed to evaluate the antioxidant activity of HDL from uremic patients. DESIGN AND METHODS: Lipoprotein properties were determined in 28 HD patients, 30 CAPD patients and 30 control subjects by spectrophotometric, chemiluminescence and electrophoresis methods. RESULTS: CAPD LDL were more resistant to oxidation than control LDL. HD and control LDL, in contrast to CAPD LDL, stimulated reactive oxygen species generation in granulocytes. The HDL ability to protect LDL against oxidation was impaired in renal patients. CONCLUSIONS: The risk of atherosclerosis development in patients with renal failure does not appear to be related to less resistance of LDL to oxidation, but rather to the decreased HDL antioxidant capacity. PMID- 16309664 TI - Interference and blood sample preparation for a pyruvate enzymatic assay. AB - BACKGROUND: To assess the severity of circulatory failure, a pyruvate enzymatic assay was performed on whole blood using lactate dehydrogenase to catalyze the conversion of pyruvate to lactate. We investigated factors related to blood sample collection and preparation that might influence the results, including the timing of blood deproteinization, temperature of sample storage, and hemolysis. METHOD: A total of 25 whole blood specimens were collected for this study. Each sample was divided into 2 parts: one stored at room temperature (RT) and another kept on ice. The samples were deproteinizied by using 8% perchloric acid (PCA) at varying times after collection; the first deproteinization was immediately after the blood was drawn (0 h), then at 1 h intervals for 6 h and also in samples kept overnight. The supernatant samples were analyzed soon after deproteinization using a COBAS Centrifugal Analyzer. In another set of samples, the blood was immediately deproteinized, and the supernatants were stored at RT and 4 degrees C and assayed for pyruvate at varying times, as above. Finally, the effect of hemolysis on the blood pyruvate enzymatic assay was also evaluated. RESULTS: When samples were stored at RT, pyruvate levels remained constant until the third h after deproteinization, when there was an approximately 13.3% increase in pyruvate concentration. When whole blood samples were kept at 4 degrees C before deproteinization, pyruvate levels were significantly reduced over time, ranging from 37.8% to 62.2% (paired t test showed a significant mean difference, P < 0.001). No significant differences in pyruvate concentration were observed in supernatant stored at either RT or 4 degrees C. Hemolysis caused a 33.7% increase in the pyruvate concentration, equivalent to 0.18 mg pyruvate per gram per deciliter of hemoglobin. CONCLUSIONS: For a pyruvate enzymatic assay, keeping a whole blood sample at RT will not cause a significant difference in the pyruvate level as long as the sample is immediately deproteinized. Whole blood samples should not be stored in an ice bath for transport, nor should hemolyzed samples be used for a blood pyruvate enzymatic assay. PMID- 16309665 TI - An evolutionary conserved role of Wnt signaling in stem cell fate decision. AB - Wnt/Frizzled/ss-catenin-based signaling systems play diverse roles in metazoan development, being involved not only in the establishment of body axes in embryogenesis but also in regulating stem cell fate in mammalian post-embryonic development. We have studied the role the canonical Wnt cascade plays in stem cell fate determination in Hydractinia, a member of the ancient metazoan phylum Cnidaria, by analyzing two key molecules in this pathway, frizzled and ss catenin, and blocking GSK-3. Generally, frizzled was expressed in cells able to divide but absent in post-mitotic, terminally differentiated cells such as nerve cells and nematocytes. Transcripts of frizzled were identified in all embryonic stages beginning with maternal transcripts in the oocyte. Following gastrulation and in the planula larva, frizzled expression concentrated in the central endodermal mass from which the first interstitial stem cells and their derivatives arise. In post-metamorphic development, high levels of frizzled transcripts were detected in interstitial stem cells. Activating downstream events of the Wnt-cascade in the post-metamorphic life phase by blocking GSK-3 with paullones induced recruitment of nematocytes and nerve cells from the pool of interstitial stem cells. Terminal differentiation was preceded by an initial burst of proliferation of frizzled-positive i-cells. In activated i-cells, ss catenin appeared in the cytoplasm, later in the nucleus. It was subsequently again observed in the cytoplasm and eventually faded out during terminal differentiation. Our results suggest an ancient role of Wnt signaling in stem cell fate determination. PMID- 16309666 TI - Cdx4 is a direct target of the canonical Wnt pathway. AB - There is considerable evidence that the Cdx gene products impact on vertebral patterning by direct regulation of Hox gene expression. Data from a number of vertebrate model systems also suggest that Cdx1, Cdx2 and Cdx4 are targets of caudalizing signals such as RA, Wnt and FGF. These observations have lead to the hypothesis that Cdx members serve to relay information from signaling pathways involved in posterior patterning to the Hox genes. Regulation of Cdx1 expression by RA and Wnt in the mouse has been well characterized; however, the means by which Cdx2 and Cdx4 are regulated is less well understood. In the present study, we present data suggesting that Cdx4 is a direct target of the canonical Wnt pathway. We found that Cdx4 responds to exogenous Wnt3a in mouse embryos ex vivo, and conversely, that its expression is down-regulated in Wnt3a(vt/vt) embryos and in embryos cultured in the presence of Wnt inhibitors. We also found that the Cdx4 promoter responds to Wnt signaling in P19 embryocarcinoma cells and have identified several putative LEF/TCF response elements mediating this effect. Consistent with these data, chromatin immunoprecipitation assays from either embryocarcinoma cells or from the tail bud of embryos revealed that LEF1 and beta catenin co-localize with the Cdx4 promoter. Taken together, these results suggest that Cdx4, like Cdx1, is a direct Wnt target. PMID- 16309667 TI - Development of midline cell types and commissural axon tracts requires Fgfr1 in the cerebrum. AB - The adult cerebral hemispheres are connected to each other by specialized midline cell types and by three axonal tracts: the corpus callosum, the hippocampal commissure, and the anterior commissure. Many steps are required for these tracts to form, including early patterning and later axon pathfinding steps. Here, the requirement for FGF signaling in forming midline cell types and commissural axon tracts of the cerebral hemispheres is examined. Fgfr1, but not Fgfr3, is found to be essential for establishing all three commissural tracts. In an Fgfr1 mutant, commissural neurons are present and initially project their axons, but these fail to cross the midline that separates the hemispheres. Moreover, midline patterning defects are observed in the mutant. These defects include the loss of the septum and three specialized glial cell types, the indusium griseum glia, midline zipper glia, and glial wedge. Our findings demonstrate that FGF signaling is required for generating telencephalic midline structures, in particular septal and glial cell types and all three cerebral commissures. In addition, analysis of the Fgfr1 heterozygous mutant, in which midline patterning is normal but commissural defects still occur, suggests that at least two distinct FGF-dependent mechanisms underlie the formation of the cerebral commissures. PMID- 16309668 TI - Nandrolone treatment decreases the alpha1B-adrenoceptor mRNA level in rat kidney, but not the density of alpha1B-adrenoceptors in cultured MDCK-D1 kidney cells. AB - We have previously shown that treatment of rats with the anabolic androgen steroid nandrolone decreased the density of alpha1B-adrenoceptors in the rat kidney [Uhlen, S., Lindblom, J., Kindlundh, A., Muhisha, P., Nyberg, F., (2003). Nandrolone treatment decreases the level of rat kidney alpha1B-adrenoceptors. Naunyn-Schmiedeberg's Arch. Pharmacol. 368, 91-98]. This effect may have been caused either by decreased de novo synthesis of alpha1B-adrenoceptors or by increased degradation of alpha1B-adrenoceptors [corrected] In the present study, we show that treatment of rats with nandrolone decreases the level of mRNA for the alpha1B-adrenoceptor in the kidneys, implying decreased synthesis of alpha1B adrenoceptors. On the other hand, nandrolone did not decrease the density of alpha1B-adrenoceptors in Madin-Darby Canine Kidney (MDCK) cells, even though the sub-cell line tested, MDCK-D1, expresses both the androgen receptor and the alpha1B-adrenoceptor. It is concluded that the regulation of alpha1B-adrenoceptor expression by anabolic androgenic steroids is intricate and cell-type specific. PMID- 16309669 TI - Inhibitory effects of a benz[f]indole-4,9-dione analog on cancer cell metastasis mediated by the down-regulation of matrix metalloproteinase expression in human HT1080 fibrosarcoma cells. AB - In our previous study, a synthetic benz[f]indole-4,9-dione analog, 2-amino-3 ethoxycarbonyl-N-methylbenz[f]indole-4,9-dione (SME-6), exhibited a potential anti-tumor activity. We, in this study, further explored the anti-metastatic and anti-invasive effect of SME-6 by determining the regulation of matrix metalloproteinases (MMPs). MMPs, zinc-dependent proteolytic enzymes, play a pivotal role in tumor metastasis by cleavage of extracellular matrix as well as non-matrix substrates. On this line, we examined the influence of SME-6 on the expressions of MMP-2, -9, membrane type 1-MMP (MT1-MMP), tissue inhibitor of metalloproteinases (TIMP-1, -2), and in vitro invasiveness of human fibrosarcoma cells. Dose-dependent suppressions of MMPs and TIMP-2 mRNA levels were observed in SME-6-treated HT1080 human fibrosarcoma cells detected by reverse transcriptase-polymerase chain reaction. TIMP-1 mRNA level, however, was induced in a dose-dependent manner. Gelatin zymographic analysis also exhibited a significant down-regulation of MMP-2 and -9 expression in HT1080 cells treated with SME-6 compared to controls. Furthermore, SME-6 inhibited the invasion, motility, and migration of tumor cells. Taken together, these data provide a possible role of SME-6 as a potential antitumor agent with the markedly inhibition of the metastatic and invasive capacity of malignant cells. PMID- 16309670 TI - Zebrafish and Xenopus tadpoles: small animal models to study angiogenesis and lymphangiogenesis. AB - Small vertebrate organisms have emerged as key players in the post-genomic era for the functional characterization of novel genes on a high-throughput scale. In this context, the zebrafish embryos and Xenopus tadpoles represent attractive and valuable models to rapidly identify and characterize novel genes involved in angiogenesis and lymphangiogenesis-a significant task with a consequent impact on the design of more effective therapeutic strategies. The advantages of these two models will be discussed in the present review. PMID- 16309671 TI - Inhibition of PI3-kinase-Akt pathway enhances dexamethasone-induced apoptosis in a human follicular lymphoma cell line. AB - Glucocorticoids are commonly used in the treatment of various lymphoid malignancies. In the present study, we show that dexamethasone (Dex) induced depolarization of mitochondrial membrane, release of cytochrome c and DNA fragmentation in a human follicular lymphoma cell line, HF28RA. New protein synthesis was required before Dex-induced mitochondrial changes, and the kinetics of the apoptotic events correlated with the upregulation of the Bim protein. Furthermore, we studied whether specific inhibitors of known survival pathways would potentiate Dex-induced apoptosis. Our results show that inhibition of PKC and ERK pathways had no effect on apoptosis. In contrast, inhibition of PI3 kinase or Akt markedly enhanced Dex-induced apoptosis. The enhancement was seen at the mitochondrial level, and the kinetics of apoptosis was notably accelerated. In addition, inhibition of PI3-kinase did not alter levels of Bax, Bcl-2, Bcl-X(L) or Bim proteins in mitochondria but caused translocation of the pro-apoptotic protein Bad to mitochondria. However, inhibition of PI3-kinase-Akt pathway and subsequent translocation of Bad to mitochondria did not induce apoptosis itself. Based on these results and our current understanding of Bim and Bad action, it seems that both proteins play a synergistic role in this process. Thus, these results indicate that inhibitors of PI3-kinase-Akt pathway might be combined in future with glucocorticoids to improve the treatment of lymphoid malignancies. PMID- 16309673 TI - Effects of blue honeysuckle (Lonicera caerulea L.) extract on lipopolysaccharide induced inflammation in vitro and in vivo. AB - The aim of the present study was to investigate the effects of blue honeysuckle extract (BHE), which contains high level of phenolic compounds, on endotoxin induced uveitis (EIU). Male Lewis rats were randomly divided into 5 groups with 14 rats in each (eight rats for collection of aqueous humor, six rats for histologic examination). EIU was induced by a footpad injection of lipopolysaccharide (LPS). 1, 10, or 100 mg of BHE was injected intravenously immediately after LPS injection. The aqueous humor was collected at 24 h after LPS injection, the number of infiltrating cells, protein concentration, nitric oxide (NO), tumor necrosis factor (TNF)-alpha, and prostaglandin (PG)-E2 levels in the aqueous humor were determined. Some eyes were enucleated for histologic examination and immunohistochemical analysis. Immunohistochemical staining with a monoclonal antibody against activated nuclear factor (NF)-kappaB was performed to evaluate the effect of BHE on NF-kappaB activation. To further clarify the anti inflammatory effect, RAW264.7 cells (a mouse macrophage cell line) were stimulated with LPS in the presence or absence of BHE and its major phenolics, cyanidin 3-glucoside (C3G), cyanidin 3-rutinoside (C3R), chlorogenic acid (CA). Expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. BHE treatment significantly reduced the inflammatory cell infiltration, the protein concentration, the levels of NO, TNF alpha and PGE2 in the aqueous humor and improved histologic status of the ocular tissue. The number of activated NF-kappaB-positive cells was lower in the iris ciliary body treated with BHE at 3 h after LPS injection. BHE significantly suppressed the production of NO, PGE2 and TNF-alpha in the culture medium as well as the expression of iNOS and COX-2 by LPS-stimulated RAW264.7 cells in a dose dependent fashion. C3G, C3R and CA showed no or weak inhibitory effects on the level of inflammatory mediators and the expression of iNOS and COX-2. These results suggest that BHE attenuates the degree of inflammation in eyes with EIU by inhibiting the NF-kappaB dependent signaling pathway and the subsequent production of proinflammatory mediators. PMID- 16309672 TI - Effect of inflammation on lacrimal gland function. AB - The lacrimal gland is the main contributor to the aqueous layer of the tear film. It secretes proteins, electrolytes and water, which helps to nourish and protect the ocular surface. Lacrimal gland secretion is primarily under neural control, which is achieved through a neural reflex arc. Stimuli to the ocular surface activate afferent sensory nerves in the cornea and conjunctiva. This in turn activates efferent parasympathetic and sympathetic nerves in the lacrimal gland to stimulate secretion. Sex steroid hormones are also important regulators of lacrimal gland functions. A decrease or lack of lacrimal gland secretion is the leading cause of aqueous tear deficient dry eye syndrome (DES). It has been suggested that DES is an inflammatory disorder that affects the ocular surface and the lacrimal gland. In several pathological instances, the lacrimal gland can become a target of the immune system and show signs of inflammation. This can result from autoimmune diseases (Sjogren's syndrome), organ transplantation (graft versus host disease), or simply as a result of aging. The hallmarks of lacrimal gland inflammation are the presence of focal lymphocytic infiltrates and increased production of proinflammatory cytokines. The mechanisms leading to lacrimal gland dysfunction are still poorly understood. Apoptosis, production of autoantibodies, hormonal imbalance, alterations in signaling molecules, neural dysfunction, and increased levels of proinflammatory cytokines have been proposed as possible mediators of lacrimal gland insufficiency in disease states. PMID- 16309674 TI - Molecular and synaptic changes in the hippocampus underlying superior spatial abilities in pre-symptomatic G93A+/+ mice overexpressing the human Cu/Zn superoxide dismutase (Gly93 --> ALA) mutation. AB - Although amyotrophic lateral sclerosis (ALS) is mainly considered as a motor disease, extramotor neural and cognitive alterations have also been reported in ALS patients. There is evidence that mutations in the Cu/Zn superoxide dismutase (SOD1) gene are implicated in about 20% of familiar ALS and transgenic mice overexpressing the human Cu/Zn superoxide dismutase (GLY(93) --> ALA) mutation show an ALS-like phenotype. However, while motor behavior has been extensively analyzed in these mutants, little is known on their cognitive abilities. To characterize the pre-symptomatic cognitive profile of G93A+/+ mice, we estimated their capability to detect spatial novelty and examined several indexes of their hippocampal function. We found an enhancement of spatial abilities in mutant mice associated with (1) a higher expression of hippocampal AMPA subunit GluR1 mRNA and of GluR1 protein levels, and (2) an increased induction and maintenance of long-term potentiation (LTP) at Schaffer collateral-CA1 synapses. Thus, before leading to extensive neuronal excitotoxicity, the high endogenous levels of glutamate present in the brain of pre-symptomatic G93A+/+ mice could mediate site specific molecular and synaptic changes providing favorable conditions to spatial information processing. These findings suggest that identification of pre symptomatic behavioral changes in murine models of ALS may point to early neural abnormalities selectively associated with mutations in the Cu/Zn superoxide dismutase (SOD1) gene. PMID- 16309675 TI - Schizophrenia--a stem cell disorder. PMID- 16309676 TI - Morris water maze search strategy analysis in PDAPP mice before and after experimental traumatic brain injury. AB - Traumatic brain injury (TBI) is a common cause of cognitive dysfunction and a major risk factor for Alzheimer's disease (AD). PDAPP mice, a transgenic line overexpressing a mutant human amyloid precursor protein (APP) implicated in familial AD, have markedly impaired behavioral performance in the Morris water maze relative to wild-type (WT) littermates. Performance further deteriorates following experimental TBI in both PDAPP and WT mice. However, the aspects of cognitive function involved are not well understood. Here, we have analyzed search strategies used in the water maze by 3-4 month old PDAPP and WT C57Bl6 littermates both before and after moderate controlled cortical impact TBI. Prior to TBI, PDAPP mice used less spatial strategies and more nonspatial systematic strategies and strategies involving repetitive looping than WT mice. With training, PDAPP mice used more spatial strategies and less repetitive looping. After TBI, PDAPP mice lost use of spatial strategies and relied more on repetitive looping. TBI in WT mice also reduced their use of spatial strategies but instead caused a switch to nonspatial systematic strategies. We also analyzed changes in the efficiency with which mice used each individual strategy, but found that differences in which strategies were used quantitatively accounted for most of the differences in performance between groups. These results demonstrate that suboptimal search strategy use in addition to effects on spatial learning and memory underlies the impaired performance of PDAPP mice and further deterioration following TBI. Human TBI patients may have analogous poor use of problem solving strategies. PMID- 16309677 TI - The beta-tubulin genes of two Strongyloides species. AB - The World Health Organization is sponsoring major treatment programs with the aim of controlling helminth infection throughout the tropical world. Prominent among the anthelmintics recommended for use in these programs are drugs in the benzimidazole (BZ) class. Resistance to these drugs has been associated with polymorphisms in the beta-tubulin gene. We have cloned and sequenced the beta tubulin genes of Strongyloides stercoralis and Strongyloides ratti and have proceeded to develop a protocol for genotyping single worms for polymorphisms in beta-tubulin. Our findings indicate that S. ratti has a single beta-tubulin gene, making DNA sequence analysis of a single larva PCR product a feasible means of studying BZ resistance in these species. Our genotyping test allows the identification of polymorphisms at codons 167, 198, and 200 in the Strongyloides beta-tubulin gene, thus enabling survey for BZ resistant genotypes. PMID- 16309678 TI - A monopartite nuclear localization sequence regulates nuclear targeting of the actin binding protein myopodin. AB - Myopodin is an actin bundling protein that shuttles between nucleus and cytoplasm in response to cell stress or during differentiation. Here, we show that the myopodin sequence 58KKRRRRARK66, when tagged to either enhanced green fluorescent protein (EGFP) or to enhanced cyan fluorescent protein-CapG (ECFPCapG), is able to target these proteins to the nucleolus in HeLa or HEK293T cells. By contrast, 58KKRR61-ECFP-CapG accumulates in the nucleus. Mutation of 58KKRRRRARK66 into alanine residues blocks myopodin nuclear import and promotes formation of cytoplasmic actin filaments. A second putative nuclear localization sequence, 612KTSKKKGKK620, displays much weaker activity in a heterologous context, and appears not to be functional in the full length protein. Thus myopodin nuclear translocation is dependent on a monopartite nuclear localization sequence. PMID- 16309679 TI - Bacteria in the oral mucosa and its effects on the measurement of cortisol, dehydroepiandrosterone, and testosterone in saliva. AB - Bacteria load in saliva was experimentally manipulated, and the consequences for the measurement of salivary testosterone (T), dehydroepiandrosterone (DHEA), and cortisol (C) were examined. Healthy adults (n = 19) donated the first saliva sample upon rising after which they rinsed their mouths with water, waited 10 min, and donated a second sample. Samples were either left untreated or passed through a 0.22-microm filter and then frozen at -80 degrees C or incubated at room temperature (RT) for 10 days. Aliquots of each sample were cultured on agar to determine baseline and post-incubation (or freezing) bacteria load. Bacteria counts were not significantly influenced by rinsing (with water), were substantially reduced by filtration, and increased by incubation at RT. Average levels of salivary T and C, but not DHEA, were significantly lower in samples stored at RT than samples frozen the day of collection. The change in bacteria count induced by storing samples at RT was associated with a change in testosterone but not cortisol or DHEA. When samples were passed through a 0.22 microm filter bacteria counts were reduced, and the association between bacteria and testosterone was reduced to non-significant. These findings contribute to a growing body of literature revealing that the process of sample collection, storage, and handling can dramatically influence the accuracy of information generated when salivary biomarkers are integrated into research and clinical diagnostics. PMID- 16309680 TI - Review of the pathophysiology and acute management of haemorrhage in pelvic fracture. AB - Mortality following pelvic fractures has declined dramatically as better methods of controlling haemorrhage, such as angioembolisation to control arterial bleeding, have been introduced. But about 10% of patients still die, despite these advances. To save these patients, the key questions in managing pelvic fractures are: which patients are at highest risk for a life-threatening bleed, in these patients, what is the exact anatomical source of the bleeding and what is the best way to stop it? There is wide consensus that bleeding is most likely to occur with unstable fractures. However, it remains difficult to predict which fractures will actually cause excessive bleeding. Current treatment protocols rely on angiographic embolisation and external fixation, either alone or in combination. Direct pelvic packing is gaining in popularity, but, ultimately, the ideal treatment method remains unclear. The purpose of this review is to examine our current understanding of the pathophysiology and management of bleeding pelvic fractures. PMID- 16309681 TI - Surgical treatment of non-unions: A case for internal fixation. AB - Non-unions present a particular challenge to the fracture surgeon and are frequently underestimated by the average practitioner. Although the use of ring fixation techniques is gaining in popularity, there remains a definite role for the use of bone grafting and plate fixation within certain clinical situations. The role of decortication, or petalling, as part of the surgical approach, is emphasized. This review addresses the basic considerations in choosing to use internal fixation to manage non-unions and outlines some of the principles of these techniques. Case examples are presented. PMID- 16309682 TI - Misoprostol in preventing postpartum hemorrhage: a meta-analysis. AB - OBJECTIVE: To assess misoprostol's ability to prevent postpartum hemorrhage (PPH) where no alternatives exist. Comparison to oxytocics demonstrates how similarly misoprostol achieves a level of effectiveness-obtainable only in hospitals-in remote locations around the world. METHOD: Using the Mantel-Haenszel fixed effects model and the DerSirmonian and Laird random-effects model, summary statistics indicated that misoprostol's excess risk of PPH was only 4% when compared to oxytocics. RESULT: This risk difference was well within the range of expected results for all uterotonic agents and does not warrant branding misoprostol as an inferior drug. CONCLUSION: Conventional uterotonic drugs should not be used to set the lowest-accepted level of effectiveness in settings where they are entirely unsuitable. Continuing to weigh the benefits of one effective drug against another only delays the distribution of misoprostol in countries where it is the only feasible choice and must be measured against no treatment at all. PMID- 16309684 TI - Gestational age determination and prevention of HIV perinatal transmission. AB - OBJECTIVE: To compare different methods of gestational age (GA) measurement for ensuring effective zidovudine (ZDV) prophylaxis to prevent mother-to-child transmission of HIV. METHODS: For 1398 HIV-infected women enrolled in a perinatal prevention trial, gestation durations were calculated based on GA estimated using ultrasound (US), date of last menstruation period (LMP), first fundal height (FH(1)), and a specific algorithm was developed to provide a "reference" GA. The performance of each GA estimate was evaluated by the percentage of women who would have received > or =8 weeks ZDV, if prophylaxis was initiated at 28 weeks. RESULTS: The performances of the algorithm, US, LMP, and FH(1) were 95.5%, 94.8%, 88.4%, and 83.7%, respectively. US and FH(1) were significantly better when estimated before and after 24 weeks, respectively. CONCLUSION: In situations where no US is available and LMP is not or imprecisely known, FH(1) can be used after 24 weeks to schedule ZDV initiation date. PMID- 16309685 TI - Social regulation, medicalisation and the nurse's role: insights from an analysis of nursing documentation. AB - BACKGROUND: Medicine is recognised as a dominant source of governmentality and social regulation, and although nursing has been implicated in the same process, analytical work in this area has been sparse. OBJECTIVES: The article aims to present an analysis of nursing records in order to understand the structural and social processes that mediate the texts. METHODS: 45 sets of nursing records drawn from four clinical sites in Ireland were subjected to a discourse analysis. RESULTS: This article focuses on two main themes that were derived from data: (i) the manner in which nurses controlled, regulated and invigilated patients' activities of daily living and (ii) the way in which activities of daily living were mediated by a biomedical worldview in the clinical settings. Through the organising framework of Activities of Daily Living (ADLs), normative social practices relating to hygiene, eating and drinking, sleeping and so forth were surveyed and monitored within clinical settings. We construct qualitative categories around a range of ways that nurses assessed and judged patients' capacities at ADLs. Furthermore, it is argued that the framework of ADLs epitomises the medicalisation of normative social practices, whereupon the most mundane of normal functions become redefined as an actual or potential clinical pathology, legitimating nursing interventions. According to the nursing documentation, biochemical interventions in the form of various medications were the most dominant means through which nurses attempted to restore or improve the functional capacity of an ADL. CONCLUSION: We conclude by proposing that nurses' invigilation of patients' ADLs is not necessarily a repressive feature of nursing practice, but rather has the potential to be used to advocate on patients' behalf in certain circumstances. PMID- 16309686 TI - Testing a conceptual model of the image of nursing in Taiwan. AB - AIM: It was believed that after identifying the differences and similarities in the perception of nurses and the public of the image of nursing, it would be important to link these perceptions to nursing care quality for practical and future implications for professionalism and the process of nursing service delivery. As a result, this cross-sectional, quantitative study tested a conceptual model of the image of nursing in Taiwan. This research investigated the perceived images and expected images of Taiwanese nurses in a sample of nurses and non-nurse subjects. The targeted dependent variable was the participants' general perception toward the image of Taiwanese nurses. METHODS: This project was conducted at a private university located in southern Taiwan, between November 2004 and January 2005. The final sample included 488 participants of undergraduate students, graduate students, university staff, or faculty members, for a general response rate of 87.6%. The study questionnaire was developed based upon previous studies. Descriptive and ordinal logistic regression analyses were performed. RESULTS: Based on the results of the four developed ordinal logistic regression models: (1) being male, (2) having positive perceptions toward the angel of mercy and (3) the careerist aspects of nursing image, (4) having negative perceptions toward the bureaucratic aspect of the nursing image, (5) having less of a difference between the participants' perceptions and their expectations toward the romantic aspect of the nursing image, and (6) being more satisfied with the professional services provided by Taiwanese nurses, explained the strength of the levels of general perception toward the image of Taiwanese nurses. DISCUSSION: To minimizing the gap between patients' expectation and perception levels of nursing services, nursing leaders should continue to monitor patients' and potential future customers' perceptions toward the image of nurses in Taiwan and work to improve that image, e.g., through on-job training to strengthen nurses' analytical skills. PMID- 16309687 TI - The evolution of European guidelines: changing the management of cholesterol levels. AB - Cardiovascular disease (CVD) is the leading cause of death in Europe and the US. This paper reviews the evolution of the Joint European Societies' guidelines with respect to their lipid recommendations. We stress the importance of lowering lipid levels to, or below, the currently recommended goals and argue that patients' global risk for CVD, rather than baseline lipid levels, should direct the intensity of lipid-lowering treatment. However, the emphasis on near-term (i.e., in the next 10 years) global risk estimation may under-emphasize the importance of considering lifetime cardiovascular risk in treatment decisions. Although various guidelines' thresholds for treatment initiation and recommended goals differ, they are similar in the theme of treating global risk. Most clinical trials have not identified a threshold of cholesterol level beyond which lowering cholesterol levels no longer provides cardiovascular benefit. An urgent call for action is needed to improve goal attainment in patients with or at risk for CVD. Improving access to risk-reducing treatments should be a priority. PMID- 16309688 TI - A novel antireflux device based on magnets. AB - BACKGROUND: The problem of eliminating gastroesophageal reflux (GER) with simple, effective and devoid of unpleasant side effects procedures is still unresolved. We tried to settle this problem with a magnetic device that should be applied to the distal end of the esophagus. MATERIALS AND METHODS: Two plastoferrite magnets of 2 x 4 x 0.5cm(1) were applied, on the opposite sides of a flaccid polyethylene tube mimicking the physical characteristics of the terminal esophagus. The two magnets attracting themselves compressed the tube, creating an artificial high pressure zone that divided the tube in two segments. Both segments of the tube were connected to pressure transducers and a polygraph and one of them was connected to a hydraulic pump. The pressure was progressively increased in this segment up to a value sufficient to detach the magnets with consequent flowing of the water in the other segment of the tube. RESULTS: The progressive increase of the pressure in a segment of the tube detached the magnets allowing a free flow into the other segment when the pressure reached an average value of 9.75+/-1.05 mmHg (mean+/-SD). CONCLUSIONS: A couple of magnets clamping a tube with the characteristics of the distal esophagus is able to prevent the passage of liquid with a pressure value near to that of a normal lower esophageal sphincter. This magnetic device could be useful to maintain closed a sphincter unable to prevent gastroesophageal reflux. PMID- 16309689 TI - Liquid chromatography determination of natural dyes in extracts from historical Scottish textiles excavated from peat bogs. AB - Textiles excavated from Scottish sites belonging now to the collections of the National Museums of Scotland, including seventeenth century textiles from peat bogs in the Scottish Highlands and Islands, were selected for analysis by high performance liquid chromatography with photodiode array detection (PDA HPLC) to detect whether any dyes remained and, if so, to identify their biological sources. Dye components were identified in 36 of the 81 samples analysed. Although it was not possible to identify the exact sources of the dyestuffs because of the wide-spread occurrence of these natural dyes components, the study has shown that textiles previously not thought to have been coloured had detectable traces of dye. Before the historical textiles were analyzed, an improved extraction procedure that combined the routine acid hydrolysis method with one using dimethylformamide (DMF) was applied. The DMF method enabled increased recovery of major flavonoid and anthraquinoid compounds, and very high efficiency of recovery of indigotin even in textiles with no colour visible, thereby complementing the acid hydrolysis method already in use. Extracts from historical thread samples were analysed by PDA HPLC using a reversed-phase gradient system comprising of a C18 column (150 mm x 4.6 mm i.d., 25 +/- 1 degrees C) with water, methanol and o-phosphoric acid at an eluent flow rate of 1.2 ml/min. A preliminary investigation to improve the detection limits further for a selection of natural dyes was made by comparing results from the 4.6mm internal diameter (i.d.) column with a narrow bore C18 column (2.1 mm i.d.). An increase in the detector response was observed for narrow-bore column proving its possibility of enhancement of sensitivity. PMID- 16309690 TI - Efficient approach for the reliable quantification and confirmation of antibiotics in water using on-line solid-phase extraction liquid chromatography/tandem mass spectrometry. AB - The potential of solid-phase extraction coupled on-line to liquid chromatography/electrospray tandem mass spectrometry (SPE-LC-ESI-MS/MS) has been investigated in this paper for the efficient sensitive quantification and confirmation of 16 antibiotics in water. The list of targeted analytes included 10 quinolones (oxolinic acid (OXO), nalidixic acid (NAL), flumequine (FLU), marbofloxacine (MAR), ofloxacine (OFLO), enrofloxacine (ENR), pefloxacine (PEF), ciprofloxacine (CIP), pipemidic acid (PIPE), norfloxacine (NOR)) and 6 penicillins (penicillin G (PEN), oxacillin (OXA), dicloxacillin (DIC), piperacillin (PIP), cloxacillin (CLO) and ampicillin (AMP)) that were determined in ground and surface water. The procedure is based on the injection of 9.8 mL of sample into the SPE-LC-MS/MS system and the measurement of antibiotics by selected reaction monitoring mode, using a triple quadrupole analyser. The method has been validated at realistic low concentrations that might be present in environmental water, i.e. 10 and 100 ng L(-1), obtaining recoveries between 74% and 123% with relative standard deviation lower than 14%. Matrix effects were not relevant in most of cases, except for ampicillin in surface water, where notable signal suppression was observed. The limits of detection were as low as 0.4-4.3 ng L(-1). The method developed allows the rapid screening and quantification of all the analytes selected by acquiring one MS/MS transition (normally the most sensitive) for each compound. It was applied to a number of actual surface and groundwater samples with several compounds being detected, mainly quinolones, at low ng L(-1) levels. Special attention was given to the confirmation of compounds detected in water due to the difficulties of obtaining confident confirmation at low ng L(-1). This matter has been of growing concern in the last few years as reflected by recent papers and correspondence. The acquisition of several MS/MS transitions for each compound detected in a second independent analysis allowed the unequivocal confirmation of identity, avoiding reporting false-positives. Finally, the potential of QTOF instruments to confirm positive samples has also been evaluated and compared with triple quadrupole analysers. PMID- 16309691 TI - Chlorine isotope fractionation of a semi-volatile organochlorine compound during preparative megabore-column capillary gas chromatography. AB - Chlorine isotope fractionation during preparative capillary gas chromatography (pcGC) was investigated using 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) as a model compound for semi-volatile organochlorine (OCl) molecules. Chlorine isotope analysis by thermal ionization mass spectrometry revealed no significant alteration of the chlorine isotope composition when the whole peaks were collected in pcGC (delta37Cl -3.2 per thousand versus -3.6 per thousand for the unprocessed DDT, +/-0.5 per thousand SD). However, distinct isotope fractionations were measured for the front (delta37Cl -5.1 per thousand) and tail (delta37Cl -1.8 per thousand) segments of partially collected samples. Isolation of individual OCls by pcGC enables accurate off-line chlorine isotope analysis, and thus facilitates the investigation of naturally occurring OCls. PMID- 16309692 TI - Ultrasound-assisted liquid-phase microextraction and high-performance liquid chromatographic determination of nitric oxide produced in PC12 cells using 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-difluoroboradiaza-s indacene. AB - To reveal the function of nitric oxide (NO) that has been reported as an intracellular second messenger and a diffusible intercellular messenger, a novel ultrasound-assisted liquid-phase microextraction (ULPME) and high-performance liquid chromatographic method for the determination of NO produced in PC12 cells has been developed. 1,3,5,7-Tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl) difluoroboradiaza-s-indacene (DAMBO-CO(2)Et) has been used for NO trapping to form corresponding triazole (DAMBO-CO(2)Et-T). The enrichment factor of DAMBO CO(2)Et-T reached 150 in 2.5 min with the proposed ULPME method. NO produced in (200+/-5) PC12 cells has been determined directly with the detection limit (S/N=3) of 2.5 x 10(-13)mol/L (5a mol), which was the lowest in HPLC determination of NO reported yet. The proposed method was sensitive, selective, convenient and rapid, which has great prospect in the qualitative and quantitative determination of NO in minimal biological samples. PMID- 16309693 TI - Development of microwave-assisted extraction followed by headspace single-drop microextraction for fast determination of paeonol in traditional Chinese medicines. AB - Paeonol is an important active component present in traditional Chinese medicines (TCMs), which was used for the treatment of many diseases such as eczema. In this work, microwave-assisted extraction (MAE) was firstly combined with headspace single-drop microextraction (HS-SDME), and applied to rapid determination of paeonol in two TCMs of Cynanchum paniculatum and Paeonia suffruticosa. In the proposed method, paeonol in TCMs was isolated by using MAE, followed by extraction and concentration by HS-SDME, and detected by gas chromatography-mass spectrometry (GC-MS). The experiment parameters of MAE and HS-SDME were discussed, and the method precision, recovery and detection limit were also studied. To further demonstrate the reliability of the quantification, both the proposed method and a standard method of steam distillation (SD) were simultaneously applied to quantitative analysis of paeonol in TCM samples from different growing areas. The experimental results show that MAE-HS-SDME is a simple and rapid method for the quantitative analysis of paeonol in TCMs, and is also a potential and alternative tool for quality monitoring for the two TCMs of C. paniculatum and P. suffruticosa. PMID- 16309694 TI - Characterization of the volatile fraction emitted by phloems of four pinus species by solid-phase microextraction and gas chromatography-mass spectrometry. AB - Pine forests constitute some of the most important renewable resources supplying timber, paper and chemical industries, among other functions. Characterization of the volatiles emitted by different Pinus species has proven to be an important tool to decode the process of host tree selection by herbivore insects, some of which cause serious economic damage to pines. Variations in the relative composition of the bouquet of semiochemicals are responsible for the outcome of different biological processes, such as mate finding, egg-laying site recognition and host selection. The volatiles present in phloem samples of four pine species, P. halepensis, P. sylvestris, P. pinaster and P. pinea, were identified and characterized with the aim of finding possible host-plant attractants for native pests, such as the bark beetle Tomicus piniperda. The volatile compounds emitted by phloem samples of pines were extracted by headspace solid-phase micro extraction, using a 2cm 50/30mm divinylbenzene/carboxen/polydimethylsiloxane table flex solid-phase microextraction fiber and its contents analyzed by high resolution gas chromatography, using flame ionization and a non polar and chiral column phases. The components of the volatile fraction emitted by the phloem samples were identified by mass spectrometry using time-of-flight and quadrupole mass analyzers. The estimated relative composition was used to perform a discriminant analysis among pine species, by means of cluster and principal component analysis. It can be concluded that it is possible to discriminate pine species based on the monoterpenes emissions of phloem samples. PMID- 16309695 TI - A novel organoclay with antibacterial activity prepared from montmorillonite and Chlorhexidini Acetas. AB - A series of novel organoclays with antibacterial activity were synthesized using Ca-montmorillonite and Chlorhexidini Acetas (CA) by ion-exchange. The resultant organoclays were characterized using X-ray diffraction (XRD), high-resolution thermogravimetric analysis (HRTG) and Fourier transform infrared spectroscopy (FTIR). Their antibacterial activity was assayed by so-called halo method. In the organoclays prepared at low CA concentration, CA ions within the clay interlayer adopt a lateral monolayer while a 'kink' state or a special state with partial overlapping of the intercalated CA in the organoclays prepared at 1.0-4.0 CEC. HRTG analysis demonstrates that CA located outside the clay interlayer exists in all synthesized organoclays, resulting from the complex molecular configuration of CA. The dramatic decrease of the surface adsorbed water and interlayer water is caused by the surface property transformation and the replacement of hydrated cations by cationic surfactant. These observations are supported by the results of FTIR. Antibacterial activity test against E. coli demonstrates that the antibacterial activity of the resultant organoclays strongly depends on the content of CA. Meanwhile, the resultant organoclay shows a long-term antibacterial activity that can last for at least one year. These novel organoclays are of potential use in synthesis of organoclay-based materials with antibacterial activity. PMID- 16309696 TI - Compression of dispersions to high stress under electric fields: effects of concentration and dispersing oil. AB - Dispersions of various concentrations (15-35%) were prepared in silicone oils of vastly different viscosities (40, 1000, 10,000, and 30,000 mPa s) and compressed to high values of stress while under an electric field of 2 kV/mm. A purpose of this study was to observe the effect of compression and E field simultaneously on these dispersions and assess predictions of most common and relevant theories. As shown, static stresses of over 1000 kPa could readily be obtained although the data presented here were held below 300 kPa to protect the load cell and equipment. The results are compared to and discussed in terms of a power law fit for stress vs gap since most theories predict such a dependence. The PL exponents fall around 3 ranges: (-2), (-3), and much less than (-3). The PL coefficients however reflect in systematic way the viscosities of the dispersing oils. The compressive stress vs strain behavior is studied with regard to particle concentration and dispersing oil viscosity. PMID- 16309697 TI - Differential roles of PKC-theta in the regulation of intracellular calcium concentration in primary T cells. AB - Activation of T lymphocytes requires protein kinase C theta (PKC-theta) and an appropriately elevated free intracellular Ca2+ concentration ([Ca2+]i). Here, we show that phorbol 12 myristate 13-acetate (PMA) inhibited Ca2+ influx in wild type but not PKC-theta-/- T cells, suggesting that PKC-theta plays a role in PMA mediated inhibition of Ca2+ influx. In contrast, T cell receptor (TCR) crosslinking in the same PKC-theta-/- T cells did result in significantly decreased [Ca2+]i compared to wild-type T cells, suggesting a positive role for PKC-theta in TCR-mediated Ca2+ mobilization. In PKC-theta-/- mice, peripheral mature T cells, but not developing thymocytes, displayed significantly decreased TCR-induced Ca2+ influx and nuclear factor of activated T cells (NFAT) translocation upon sub-optimal TCR crosslinking. The decreased intracellular free Ca2+ was due to changes in Ca2+ influx but not efflux, as observed in extracellular and intracellular Ca2+ mobilization studies. However, these differences in Ca2+ influx and nuclear factor of activated T cells (NFAT) translocation disappeared with increasing intensity of TCR crosslinking. The enhancing effect of PKC-theta on Ca2+ influx is not only dependent on the strength of TCR crosslinking but also on the developmental stage of T cells. The underlying mechanism involved phospholipase Cgamma1 activation and inositol triphosphate production. Furthermore, knockdown of endogenous PKC-theta expression in Jurkat cells resulted in significant inhibition of TCR-induced activation of NFAT, as evidenced from NFAT reporter studies. Forced expression of a constitutively active form of calcineurin in PKC-theta-/- Jurkat cells could readily overcome the above inhibition. Thus, PKC-theta can both positively and negatively regulate the Ca2+ influx that is critical for NFAT activity. PMID- 16309698 TI - Structure and catalytic properties of an engineered heterodimer of enolase composed of one active and one inactive subunit. AB - Enolase is a dimeric enzyme that catalyzes the interconversion of 2-phospho-D glycerate and phosphoenolpyruvate. This reversible dehydration is effected by general acid-base catalysis that involves, principally, Lys345 and Glu211 (numbering system of enolase 1 from yeast). The crystal structure of the inactive E211Q enolase shows that the protein is properly folded. However, K345 variants have, thus far, failed to crystallize. This problem was solved by crystallization of an engineered heterodimer of enolase. The heterodimer was composed of an inactive subunit that has a K345A mutation and an active subunit that has N80D and N126D surface mutations to facilitate ion-exchange chromatographic separation of the three dimeric species. The structure of this heterodimeric variant, in complex with substrate/product, was obtained at 1.85 A resolution. The structure was compared to a new structure of wild-type enolase obtained from crystals belonging to the same space group. Asymmetric dimers having one subunit exhibiting two of the three active site loops in an open conformation and the other in a conformation having all three loops closed appear in both structures. The K345A subunit of the heterodimer is in the loop-closed conformation; its Calpha carbon atoms closely match those of the corresponding subunit of wild-type enolase (root-mean-squared deviation of 0.23 A). The kcat and kcat/Km values of the heterodimer are approximately half those of the N80D/N126D homodimer, which suggests that the subunits in solution are kinetically independent. A comparison of enolase structures obtained from crystals belonging to different space groups suggests that asymmetric dimers can be a consequence of the asymmetric positioning of the subunits within the crystal lattice. PMID- 16309699 TI - Structural evidence for adaptive ligand binding of glycolipid transfer protein. AB - Glycolipids participate in many important cellular processes and they are bound and transferred with high specificity by glycolipid transfer protein (GLTP). We have solved three different X-ray structures of bovine GLTP at 1.4 angstroms, 1.6 angstroms and 1.8 angstroms resolution, all with a bound fatty acid or glycolipid. The 1.4 angstroms structure resembles the recently characterized apo form of the human GLTP but the other two structures represent an intermediate conformation of the apo-GLTPs and the human lactosylceramide-bound GLTP structure. These novel structures give insight into the mechanism of lipid binding and how GLTP may conformationally adapt to different lipids. Furthermore, based on the structural comparison of the GLTP structures and the three dimensional models of the related Podospora anserina HET-C2 and Arabidopsis thaliana accelerated cell death protein, ACD11, we give structural explanations for their specific lipid binding properties. PMID- 16309700 TI - Co-axial association of recombinant eye lens aquaporin-0 observed in loosely packed 3D crystals. AB - Aquaporin-0 (AQP0) is the major membrane protein in vertebrate eye lenses. It has been proposed that AQP0 tetramers mediate contact between membranes of adjacent lens fiber cells, which would be consistent with the extraordinarily narrow inter cellular spacing. We have obtained 3D crystals of recombinant bovine AQP0 that diffract to 7.0 A resolution. The crystal packing was determined by molecular replacement and shows that, within the cubic lattice, AQP0 tetramers are associated head-to-head along their 4-fold axes. Oligomeric states larger than the tetramer were also observed in solution by native gel electrophoresis and analytical ultracentrifugation methods. In the crystals, there are no direct contacts between octamers, and it can thus be inferred that crystalline order is mediated solely by the detergent belts surrounding the membrane protein. Across the tetramer-tetramer interface, extracellular loops A and C interdigitate at the center and the perimeter of the octamer, respectively. The octamer structure is compared with that of the recently determined structure of truncated ovine AQP0 derived from electron diffraction of 2D crystals. Intriguingly, also in these crystals, octamers are observed, but with significantly different relative tetramer-tetramer orientations. The interactions observed in the loosely packed 3D crystals reported here may in fact represent an in vivo association mode between AQP0 tetramers from juxtaposed membranes in the eye lens. PMID- 16309701 TI - Designing thermostable proteins: ancestral mutants of 3-isopropylmalate dehydrogenase designed by using a phylogenetic tree. AB - We have recently developed a new method for designing thermostable proteins using phylogenetic trees of enzymes. In this study, we investigated a method for designing proteins with improved stability using 3-isopropylmalate dehydrogenase (IPMDH) from Thermus thermophilus as a model enzyme. We designed 12 mutant enzymes, each having an ancestral amino acid residue that was present in the common ancestor of Bacteria and Archaea. At least six of the 12 ancestral mutants tested showed thermal stability higher than that of the original enzyme. The results supported the hyperthermophilic universal ancestor hypothesis. The effect of ancestral residues on IPMDHs of several organisms and on the related enzyme isocitrate dehydrogenase was summarised and analysed. The effect of an ancestral residue on thermostability did not depend on the degree of conservation of the residue at the site, suggesting that the stabilisation of these mutant proteins is not related to sequence conservation but to the antiquity of the introduced residues. The results suggest also that this method could be an efficient way of designing mutant enzymes with higher thermostability based only on the primary structure and a phylogenetic tree. PMID- 16309702 TI - Roles of N-terminal pyroglutamate in maintaining structural integrity and pKa values of catalytic histidine residues in bullfrog ribonuclease 3. AB - Many proteins and bioactive peptides contain an N-terminal pyroglutamate residue (Pyr1). This residue reduces the susceptibility of the protein to aminopeptidases and often has important functional roles. The antitumor ribonuclease RC-RNase 3 (RNase 3) from oocytes of Rana catesbeiana (bullfrog) is one such protein. We have produced recombinant RNase 3 containing the N-terminal Pyr1 (pRNase 3) and found it to be indistinguishable from the native RNase 3 by mass spectrometry and a variety of other biochemical and immunological criteria. We demonstrated by NMR analysis that the Pyr1 of pRNase 3 forms hydrogen bonds with Lys9 and Ile96 and stabilizes the N-terminal alpha-helix in a rigid conformation. In contrast, the N terminal alpha-helix becomes flexible and the pKa values of the catalytic residues His10 and His97 altered when Pyr1 formation is blocked by an extra methionine at the N terminus in the recombinant mqRNase 3. Thus, our results provide a mechanistic explanation on the essential role of Pyr1 in maintaining the structural integrity, especially at the N-terminal alpha-helix, and in providing the proper environment for the ionization of His10 and His97 residues for catalysis and cytotoxicity against HeLa cells. PMID- 16309703 TI - Structural basis for transcription-coupled repair: the N terminus of Mfd resembles UvrB with degenerate ATPase motifs. AB - The transcription repair coupling factor Mfd removes stalled RNA polymerase from DNA lesions and links transcription to UvrABC-dependent nucleotide excision repair in prokaryotes. We report the 2.1A crystal structure of the UvrA-binding N terminus (residues 1-333) of Escherichia coli Mfd (Mfd-N). Remarkably, Mfd-N reveals a fold that resembles the three N-terminal domains of the repair enzyme UvrB. Domain 1A of Mfd adopts a typical RecA fold, domain 1B matches the damage binding domain of the UvrB, and domain 2 highly resembles the implicated UvrA binding domain of UvrB. However, Mfd apparently lacks a functional ATP-binding site and does not contain the DNA damage-binding motifs of UvrB. Thus, our results suggest that Mfd might form a UvrA recruitment factor at stalled transcription complexes that architecturally but not catalytically resembles UvrB. PMID- 16309704 TI - Epitope diversity of hepatitis B virus capsids: quasi-equivalent variations in spike epitopes and binding of different antibodies to the same epitope. AB - To investigate the range of antigenic variation of HBV capsids, we have characterized the epitopes for two anti-capsid antibodies by cryo-electron microscopy and image reconstruction of Fab-labeled capsids to approximately 10A resolution followed by molecular modeling. Both antibodies engage residues on the protruding spikes but their epitopes and binding orientations differ. Steric interference effects limit maximum binding to approximately 50% average occupancy in each case. However, the occupancies of the two copies of a given epitope that are present on a single spike differ, reflecting subtle distinctions in structure and hence, binding affinity, arising from quasi-equivalence. The epitope for mAb88 is conformational but continuous, consisting of a loop-helix motif (residues 77-87) on one of the two polypeptide chains in the spike. In contrast, the epitope for mAb842, like most conformational epitopes, is discontinuous, consisting of a loop on one polypeptide chain (residues 74-78) combined with a loop-helix element (residues 78-83) on the other. The epitope of mAb842 is essentially identical with that previously mapped for mAb F11A4, although the binding orientations of the two monoclonal antibodies (mAbs) differ, as do their affinities measured by surface plasmon resonance. From the number of monoclonals (six) whose binding had to be characterized to give the first duplicate epitope, we estimate the total number of core antigen (cAg) epitopes to be of the order of 20. Given that different antibodies may share the same epitope, the potential number of distinct anti-cAg clones should be considerably higher. The observation that the large majority of cAg epitopes are conformational reflects the relative dimensions of a Fab (large) and the small size and close packing of the motifs that are exposed and accessible on the capsid surface. PMID- 16309705 TI - Effect of hsp70 chaperone on the folding and misfolding of polypeptides modeling an elongating protein chain. AB - Virtually nothing is known about the interaction of co-translationally active chaperones with nascent polypeptides and the resulting effects on peptide conformation and folding. We have explored this issue by NMR analysis of apomyoglobin N-terminal fragments of increasing length, taken as models for different stages of protein biosynthesis, in the absence and presence of the substrate binding domain of Escherichia coli Hsp70, DnaK-beta. The incomplete polypeptides misfold and self-associate under refolding conditions. In the presence of DnaK-beta, however, formation of the original self-associated species is completely or partially prevented. Chaperone interaction with incomplete protein chains promotes a globally unfolded dynamic DnaK-beta-bound state, which becomes folding-competent only upon incorporation of the residues corresponding to the C-terminal H helix. The chaperone does not bind the full-length protein at equilibrium. However, its presence strongly disfavors the kinetic accessibility of misfolding side-routes available to the full-length chain. This work supports the role of DnaK as a "holder" for incomplete N-terminal polypeptides. However, as the chain approaches its full-length status, the tendency to intramolecularly bury non-polar surface efficiently outcompetes chaperone binding. Under these conditions, DnaK serves as a "folding enhancer" by supporting folding of a population of otherwise folding-incompetent full-length protein chains. PMID- 16309706 TI - Associations in the longitudinal course of body dysmorphic disorder with major depression, obsessive-compulsive disorder, and social phobia. AB - Body dysmorphic disorder (BDD) is an impairing and relatively common disorder that has high comorbidity with certain Axis I disorders. However, the longitudinal associations between BDD and comorbid disorders have not previously been examined. Such information may shed light on the nature of BDD's relationship to putative "near-neighbor" disorders, such as major depression, obsessive-compulsive disorder (OCD), and social phobia. This study examined time varying associations between BDD and these comorbid disorders in 161 participants over 1-3 years of follow-up in the first prospective longitudinal study of the course of BDD. We found that BDD had significant longitudinal associations with major depression--that is, change in the status of BDD and major depression was closely linked in time, with improvement in major depression predicting BDD remission, and, conversely, improvement in BDD predicting depression remission. We also found that improvement in OCD predicted BDD remission, but that BDD improvement did not predict OCD remission. No significant longitudinal associations were found for BDD and social phobia (although the results for analyses of OCD and social phobia were less numerically stable). These findings suggest (but do not prove) that BDD may be etiologically linked to major depression and OCD, i.e., that BDD may be a member of both the putative OCD spectrum and the affective spectrum. However, BDD does not appear to simply be a symptom of these comorbid disorders, as BDD symptoms persisted in a sizable proportion of subjects who remitted from these comorbid disorders. Additional studies are needed to elucidate the nature of BDD's relationship to commonly co occurring disorders, as this issue has important theoretical and clinical implications. PMID- 16309707 TI - Safety halls--an evaluation. AB - PROBLEM: In most countries, drivers licensing systems usually include teaching some aspects of using safety equipment (e.g., airbags and seat belts). However, there is now evidence worldwide that such education is inadequate, as indicated by, for example, the overrepresentation of young drivers who do not use seat belts. METHOD: A randomized controlled study was conducted in Sweden to evaluate the effects of visiting a facility known as a "safety hall" in combination with the mandatory skid training. The results were assessed to determine the effects of the knowledge and attitudes of learner drivers in the following subjects: airbags, securing loads, seat belts, sitting posture, speed, and tires. An experimental group and a control group comprising 658 and 668 learners, respectively, answered identical questionnaires on three different occasions (pretest, posttest 1, and posttest 2). RESULTS: The results show that, for most of the topics considered, knowledge and attitudes in both groups were better at posttest 2 than at the pretest, and in general, the best knowledge and attitudes were found in the experimental group. The combined safety/skid training seems to have had the greatest effect on seat belts and loads. The findings also indicate that the safety halls can be further improved to achieve an even better effect. IMPACT ON TRAFFIC SAFETY: The use of safety halls has improved the knowledge and attitudes of learner drivers concerning several important areas related to traffic safety. Since knowledge and attitudes are important predictors of behavior, implementing safety halls can be expected to lead to improvements, especially regarding the use of safety belts and securing loads. PMID- 16309708 TI - Gastroprotective and ulcer healing effect of ferruginol in mice and rats: assessment of its mechanism of action using in vitro models. AB - The gastroprotective activity of the diterpene ferruginol isolated from Prumnopitys andina wood and bark was determined on HCl/EtOH-induced gastric lesions in mice. The effect of the compound on the healing of subacute gastric lesions in rats was also studied. The mode of action of the diterpene was assessed using human gastric epithelial cells (AGS) and MRC-5 fibroblasts. The effect of ferruginol on the prostaglandin E2 content, protection against sodium taurocholate induced-damage and reduced glutathione content was evaluated on AGS cells as well as on the growth of AGS and fibroblast cultures. The free radical scavenging effect of ferruginol was assessed by the 1,1-diphenyl-2-picryl hydrazil radical and superoxide anion assays. The effect of ferruginol on human erythrocyte membrane lipoperoxidation was determined. The cytotoxicity of the compound was assessed by means of the neutral red uptake. At 25 mg/kg, ferruginol inhibited the appearance of gastric lesions by 60% showing similar effects than lansoprazole at 20 mg/kg. Additionally, the compound displayed a significant ulcer healing activity in rats at 25 and 50 mg/kg with curative ratios of 36.0% and 92.5%, respectively, while the reference compound ranitidine at 50 mg/kg showed a curative ratio of 79.6%. At 6 and 12 microM, ferruginol increased significantly the prostaglandin E2 content. A strong inhibition of lipoperoxidation was found (IC50: 1.4 microM), but no effect was observed on the sodium taurocholate induced-damage or reduced glutathione content. Ferruginol stimulated cell proliferation at 1-2 microM in AGS cells and at 4-8 microM in fibroblasts, with cytotoxicities (IC50) of 24 and 26 microM, respectively. Our results support that ferruginol acts as gastroprotective increasing the PGs content, protecting the cells against lipid peroxidation and improving the gastric ulcer healing by a stimulating effect on the cell proliferation. These findings encourage further pharmacological studies of ferruginol as a potential new anti-ulcerogenic drug. PMID- 16309709 TI - Prostaglandin E2 deteriorates N-methyl-D-aspartate receptor-mediated cytotoxicity possibly by activating EP2 receptors in cultured cortical neurons. AB - The activation of glutamate receptors, particularly N-methyl-D-aspartate (NMDA) receptors, initiates ischemic cascade in the early stages of cerebral ischemia. Postischemia, cerebral ischemia is also associated with an inflammatory reaction that contributes to tissue damage. The up-regulation of neuronal cyclooxygenase-2 (COX-2) and elevation of prostaglandin E2 (PGE2) have been reported to occur after cerebral ischemic insult. We therefore studied whether the COX-2 reaction product PGE2 affects glutamate receptor-mediated cell death in cultured rat cortical cells. PGE2 was found to augment NMDA-mediated cell death. The transcription of EP1, EP2, EP3 and EP4 PGE2 receptor genes was investigated using reverse transcriptase-polymerase chain reaction (RT-PCR). EP1, EP2 and EP3 receptor genes were found in cortical cells. Butaprost (an EP2 agonist) markedly enhanced NMDA-mediated cell death, whereas 17-phenyl trinor-PGE2 (an EP1 agonist) and sulprostone (an EP3 agonist) had little effect. Both PGE2 and butaprost elevated cAMP intracellular levels in the cortical cells; moreover, forskolin, an activator of adenylate cyclase, enhanced NMDA-mediated cell death. These results suggest that PGE2, acting via EP2 receptors, aggravates excitotoxic neurodegeneration by a cAMP-dependent mechanism. PMID- 16309710 TI - Antiviral effects of sophoridine against coxsackievirus B3 and its pharmacokinetics in rats. AB - Coxsackievirus B3 (CVB3) is a major pathogen for acute and chronic viral myocarditis. The aim of this study was to investigate the antiviral effects of sophoridine, an alkaloid extracted from Chinese medicinal herb, Sophora flavescens, against CVB3, and the underlying pharmacokinetics. First, we determined the antiviral effects of sophoridine against CVB3 in in vitro (primarily cultured myocardial cells), in vivo (BALB/c mice) and serum pharmacological experiments. Then, we determined the pharmacokinetic behavior in serum samples of SD rats after oral administration by HPLC. Finally, we determined the effects of sophoridine on the production of cytokines in a murine viral myocarditis model by measuring mRNA expression of some important cytokines in hearts of infected BALB/c mice by RT-PCR. We found that sophoridine exhibited obvious antiviral effects both in vitro and in vivo, and serum samples obtained from rats with oral administration of sophoridine reduced the virus titers in infected myocardial cells. The serum concentration profile correlated closely with antiviral activity profile. Moreover, sophoridine significantly enhanced mRNA expression of IL-10 and IFN-gamma, but decreased TNF-alpha mRNA expression. In conclusion, sophoridine possesses antiviral activities against CVB3, by regulating cytokine expression, and it is likely that sophoridine itself, not its metabolites, is mainly responsible for the antiviral activities. Therefore, sophoridine may represent a potential therapeutic agent for viral myocarditis. PMID- 16309711 TI - Up-regulation of integrin-linked kinase activity in rat mesangioproliferative glomerulonephritis. AB - This study investigated whether integrin-linked kinase (ILK) is involved in the pathogenesis of chronic glomerulonephritis (GN) by analyzing the expression and activity of glomerular ILK in a chronic rat model of mesangioproliferative GN. Double immunostaining of kidneys obtained at different time points with glomerular cell-specific markers revealed that ILK was primarily expressed by glomerular epithelial cells, and weakly by mesangial cells (MCs) and endothelial cells in control rats, but dramatically increased in a typical mesangial pattern at days 21 and 28 of GN. Semiquantitative assessment indicated that the level of glomerular ILK expression closely parallels the level of accumulation of glomerular extracellular matrix (ECM) as well as fibronectin (FN). Immunoprecipitation and kinase activity assays using isolated nephritic glomeruli indicated a striking increase of ILK activity on days 21 and 28 of GN. Further, cultured rat MCs overexpressing kinase-deficient ILK diminished FN assembly and collagen matrix remodeling as compared with control transfectants. The results showed that glomerular ILK expression and activity are markedly increased in an experimental model of chronic GN. Increased activity of ILK in MCs may contribute to the development of chronic mesangial alterations leading to glomerular scarring. PMID- 16309712 TI - Carrier-mediated processes at several rat brain interfaces determine the neuropharmacokinetics of morphine and morphine-6-beta-D-glucuronide. AB - We investigated whether capacity-limited transport processes were involved in morphine and morphine-6-beta-D-glucuronide (M6G) neuropharmacokinetics, at the level of the blood-brain barrier (BBB), the brain extra- and intra-cellular fluids (bECF/bICF), and the bECF/cerebrospinal fluid (CSF) interfaces. We performed transcortical retrodialysis in the rat, by perfusing morphine or M6G through the microdialysis probe in the presence or absence of probenecid. We measured for each compound the in vitro and in vivo (R(D)) probe recoveries. The in vivo R(D), which takes into account the permeability of the tissue surrounding the probe, informs about the morphine and M6G distribution capabilities from bECF to adjacent fluids (bICF, CSF, plasma). We also measured plasma and CSF concentrations at three time points after having added probenecid or not. Finally, we tested several pharmacokinetic models, assuming first-order or capacity-limited processes at each brain interface, to describe experimental morphine and M6G concentrations previously obtained in rat plasma and brain fluids. We found that morphine distributes more easily outside bECF than M6G. Adding probenecid caused a 2-fold decrease and a 1.3-fold increase in morphine and M6G R(D), respectively, and 30 min after adding probenecid, plasma and CSF concentrations increased for M6G but not for morphine. The pharmacokinetic model that gave the best fit included capacity-limited processes at the BBB and bECF/bICF interface for morphine and at the BBB and bECF/CSF interface for M6G. In conclusion, morphine accumulates into brain cells thanks to a probenecid sensitive transporter located at the bECF/bICF interface, whereas M6G is trapped in bECF thanks to transporters located at the BBB and the bECF/CSF interface. PMID- 16309713 TI - Apparent discrepancy in contamination history of a sub-tropical estuary evaluated through 210Pb profile and chronostratigraphical markers. AB - Zn and Cd concentrations, stable lead isotopes and 210Pb-derived chronology were determined in a sediment core sampled at Sepetiba Bay (South-eastern Brazil). During the last decades, the bay's watershed has been modified by the increase of industrial activities and human interventions. In particular, Zn and Cd ore treatment plants were built near the coast in 1960 and 1970, respectively, and water has been diverted from the adjacent Paraiba do Sul River watershed since 1950. The core collected at shallow depth near the industrial area exhibits four successive events: (i) at 50 cm depth, a change in the 206Pb/207Pb ratio from about 1.162 to more than 1.18 might be the result of the Sao Francisco Channel opening and water diversion from Paraiba do Sul river; (ii) at 40 cm depth, Zn concentration starts to increase (up to 0.8 mg g(-1)) (iii) above 30 cm depth, relatively high Cd concentrations (up to 1.6 microg g(-1)) are observed and (iv) at 16 cm depth, change in unsupported 210Pb slope is probably related to a waste dam built to prevent strong metal contamination in the bay. Sediment accumulation rates evaluated by Zn and Cd profiles used as time-markers are higher than those calculated from 210Pb-based chronology models. Using the constant initial concentration (CIC) model both events are supposed to date back to about 1884 and 1902, respectively, while using the constant rate of supply (CRS) model it shifts to about 1925 and 1935. Such discrepancies are probably assigned to the fact that these models do not take into account site-specific local sedimentation dynamics. In the study area, particles deposition seems to be controlled by enrichment with unsupported 210Pb transported by runoff from a mangrove flat bank. Chronology derived from a model that assumes an exponential increase of the initial 210Pb activity fits well with the estimated rates obtained from historical events. PMID- 16309714 TI - Scientific management of Mediterranean coastal zone: a hybrid ocean forecasting system for oil spill and search and rescue operations. AB - The oil spill from Prestige tanker showed the importance of scientifically based protocols to minimize the impacts on the environment. In this work, we describe a new forecasting system to predict oil spill trajectories and their potential impacts on the coastal zone. The system is formed of three main interconnected modules that address different capabilities: (1) an operational circulation sub system that includes nested models at different scales, data collection with near real time assimilation, new tools for initialization or assimilation based on genetic algorithms and feature-oriented strategic sampling; (2) an oil spill coastal sub-system that allows simulation of the trajectories and fate of spilled oil together with evaluation of coastal zone vulnerability using environmental sensitivity indexes; (3) a risk management sub-system for decision support based on GIS technology. The system is applied to the Mediterranean Sea where surface currents are highly variable in space and time, and interactions between local, sub-basin and basin scale increase the non-linear interactions effects which need to be adequately resolved at each one of the intervening scales. Besides the Mediterranean Sea is a complex reduced scale ocean representing a real scientific and technological challenge for operational oceanography and particularly for oil spill response and search and rescue operations. PMID- 16309716 TI - CYP2B6 is expressed in African Green monkey brain and is induced by chronic nicotine treatment. AB - CYP2B6 is a drug-metabolizing enzyme expressed in human tissues that can activate bupropion (a smoking cessation drug) and tobacco smoke nitrosamines and can inactivate drugs such as nicotine. Smokers have higher brain CYP2B6 protein levels compared to non-smokers but the cause of this elevation is unknown. We investigated the basal expression and the effect of chronic nicotine treatment on CYP2B6 protein in African Green monkey (Cercopithecus aethiops) brain. Basal expression of brain CYP2B6 was strong in specific cells such as the frontal cortical pyramidal cells, the cerebellar Purkinje cells and the neurons in the substantia nigra. Basal CYP2B6 protein levels varied 2.7-fold (non-significant) among 12 brain regions. All monkeys were given a subcutaneous 0.1 mg/kg nicotine test dose prior to treatment and the maximum plasma concentration achieved was 87 +/- 69 ng/ml and the half-life was 2.6 +/- 1.5 h. Monkeys were treated subcutaneously twice daily with nicotine at 0.05 mg/kg for 2 days, 0.15 mg/kg for 2 days followed by 0.3 mg/kg for 18 days (n = 6) or saline (n = 6). Chronic nicotine treatment induced CYP2B6 expression in specific cells such as astrocytes and neurons in the frontal cortex, caudate, thalamus and hippocampus. CYP2B6 protein levels were induced 1.5-fold in the frontal cortex (p < 0.01). Hepatic CYP2B6 expression was not altered by nicotine. In conclusion, CYP2B6 protein is expressed in specific cells in monkey brain and is induced by chronic nicotine treatment which may impact central metabolism of CYP2B6 substrates such as bupropion and nicotine. PMID- 16309717 TI - ERbeta mediates the estradiol increase of D2 receptors in rat striatum and nucleus accumbens. AB - Estradiol was previously reported to increase striatal D(2) receptor density. The following experiments investigated the contribution of each estrogen receptor in estradiol modulation of D(2) receptors. Ovariectomized Sprague-Dawley rats were treated for 2 weeks with an agonist for ERalpha, 4,4',4''-(4-propyl-[1H]-pyrazole 1,3,5-triyl)trisphenol (PPT), an agonist for ERbeta, 2,3-bis(4-hydroxyphenyl) propionitrile (DPN) and compared to estradiol treatment. Ovariectomy decreased D(2) agonist and antagonist striatal binding sites, specific binding was measured using [(3)H]quinpirole and [(3)H]spiperone. Estradiol prevented this decrease, while DPN but not PPT mimicked the estradiol increase of D(2) receptor specific binding. In the nucleus accumbens, ovariectomy decreased [(3)H]quinpirole specific binding in the core and left the shell unchanged. Similarly, estradiol and DPN but not PPT prevented this decrease. Neither ovariectomy nor treatments affected [(3)H]spiperone specific binding in this area. In the olfactory tubercle, neither ovariectomy nor treatments changed D(2) receptor binding. Finally, both ovariectomy and treatments did not affect D(2L), D(2S) mRNA and D(2L)/D(2S) ratios measured by semi-quantitative RT-PCR. The present results show, for the first time, that an ERbeta agonist treatment modulates D(2) receptors and suggest that ERbeta is involved in the estradiol modulation of D(2) receptors. PMID- 16309718 TI - Unique patterns of FOS, phospho-CREB and BrdU immunoreactivity in the female rat brain following chronic stress and citalopram treatment. AB - Affective disorders are common psychiatric illnesses characterized by marked gender-related prevalence. Recent evidence links chronic stress and dysregulation of neurotrophin signaling with the development of depression, while novel theories suggest that antidepressants may act by promoting intracellular adaptations linked to neuroplasticity. Although selective serotonin reuptake inhibitors (SSRIs) efficaciously improve a variety of dysfunctions in males, their neuroendocrine effects and intracellular signaling patterns in females are not well determined. Here we show that chronic footshock stress (21 days) promotes HPA axis hyperactivity (as seen by the increased FOS-ir in the paraventricular hypothalamic nucleus (PVN), plasma corticosterone and adrenal hypertrophy), reduces hippocampal BrdU immunoreactivity and suppresses cortical limbic CREB phosphorylation in female rats. Long-term citalopram treatment, in contrast, attenuates stress-induced elevation of corticosterone levels and adrenal hypertrophy, although it does not reverse footshock-mediated induction of FOS-ir in the PVN, inhibition of CREB phosphorylation and reduction of hippocampal BrdU-labeling. Moreover, citalopram administration was also associated with significant hypophagic effects and inhibition of CREB phosphorylation. These data suggest that, in female rats, normalization of chronic stress-induced HPA axis abnormalities may represent an initial phase of citalopram-mediated therapeutic actions and despite this SSRI's apparent lack of effects on neuroplasticity, we cannot exclude the possibility that some neurochemical adaptations occur in a later stage which may require more than 3 weeks of treatment to manifest. PMID- 16309719 TI - Effect of repeated exposures to cold on cognitive performance in humans. AB - The effects of repeated exposure to cold temperature on cognitive performance were examined in 10 male subjects who were exposed to control (25 degrees C) and cold (10 degrees C) conditions on 10 successive days. A cognitive test battery (ANAM-ICE) was administered each day to assess complex and simple cognitive functioning accuracy, efficiency and response time. Rectal (T(rect)) and skin temperatures, thermal sensations, metabolic rate (M) and cardiovascular reactivity were also recorded. With the used cold exposure, inducing cold sensations and discomfort, superficial skin cooling (6-7 degrees C) and a slightly lowered T(rect) (0.4 degrees C) we observed three distinct patterns of cognitive performance: 1) negative, reflected in increased response times and decreased accuracy and efficiency; 2) positive, reflected in decreased response time and increased efficiency; and 3) mixed, reflected in a pattern of increases in both accuracy and response time and decreases in efficiency, and a pattern of decreases in both accuracy and response time. T(rect), thermal sensations, diastolic blood pressure (DBP) and heart rate (HR) were independent predictors of decreased accuracy, but also decreased response time. Cognitive performance efficiency was significantly improved and response times shorter over the 10-d period both under control and cold exposures suggesting a learning effect. However, the changes in cognitive performance over the 10-d period did not differ markedly between control and cold, indicating that the changes in the thermal responses did not improve performance. The results suggest that cold affects cognitive performance negatively through the mechanisms of distraction and both positively and negatively through the mechanism of arousal. PMID- 16309720 TI - Detection of uronic oxidase activity in ripening peaches. AB - Uronic acid oxidase activity was found in an extract from harvested peaches that was incubated with citrus pectin at pH 8.5. The product of this reaction was identified by GC-MS analysis to be galactaric acid. The reaction was linear at 37 degrees C for up to 20 h, and the pH optimum was 8.5. The activity found in firm peaches one day after harvest did not change as the peaches softened over 5 days to eating softness. The incubation conditions were those suitable for monitoring the activity of pectate lyase, but instead of finding an increase in galacturonosyl residue reducing groups due to generation of pectin-derived oligosaccharides, uronic acid oxidase catalyzed the oxidation of the aldehyde reducing functions to carboxyl groups. PMID- 16309722 TI - In vitro anti-inflammatory activities of new steroidal antedrugs: [16alpha,17alpha-d] Isoxazoline and [16alpha,17alpha-d]-3'-hydroxy-iminoformyl isoxazoline derivatives of prednisolone and 9alpha-fluoroprednisolone. AB - A series of new anti-inflammatory steroidal antedrugs with C-16,17-isoxazoline ring system were synthesized and their pharmacological activities were evaluated. We reported earlier that these compounds are promising antedrugs based on the results of 5-day rat croton oil ear edema assay. In the present study, most of these compounds showed high binding affinities to the glucocorticoid receptor of liver cytosol. 21-acetyloxy-9alpha-fluoro-11beta-hydroxy-3,20-dioxo-1,4 pregnadieno [16alpha,17alpha-d] isoxazoline (FP-ISO-21AC) and 11beta,21-dihydroxy 9alpha-fluoro-3,20-dioxo-1,4-pregnadieno [16alpha,17alpha-d] isoxazoline (FP-ISO 21OH) were found 5.0-, 5.3-fold more potent than prednisolone, respectively. Inhibitory effects of the antedrugs on the nitric oxide (NO) production were assessed using LPS-stimulated RAW 264.7 murine macrophage cells. All these steroidal antedrugs exhibited concentration-dependent inhibition of NO production, but their relative potencies were lower than prednisolone. In vitro metabolism study in rat plasma showed that FP-ISO-21AC and 21-acetyloxy-9alpha fluoro-11beta-hydroxy-3,20-dioxo-1,4-pregnadieno [16alpha,17alpha-d]-3' hydroxyiminoformyl isoxazoline (FP-OXIM-21AC) were hydrolyzed rapidly, with the half-lives of 2.1 and 4.2 min, respectively. The half-lives of FP-ISO-21OH and 11beta,21-dihydroxy-9alpha-fluoro-3,20-dioxo-1,4-pregnadieno [16alpha,17alpha-d] 3'-hydroxyiminoformyl isoxazoline (FP-OXIM-21OH) were 92.2 and 110.2 min, respectively. PMID- 16309723 TI - Biological activities of a lectin from Bothrops jararacussu snake venom. AB - Snake venoms contain saccharide-binding lectins. In this work, we examined the biological activities of a lectin (BjcuL) purified from Bothrops jararacussu snake venom by chromatography on non-derivatized Sepharose 4B and Sephacryl S-200 HR. The protein, a homodimer with subunits of 14.5 kDa, gave a single immunoprecipitin line in immunoelectrophoresis and cross-reacted in ELISA with antivenoms raised against Bothrops spp. (lanceheads), Micrurus spp. (coral snakes), Crotalus durissus terrificus (South American rattlesnake), and arthropod (Loxosceles gaucho, Phoneutria nigriventer and Tityus serrulatus) venoms. BjcuL agglutinated human formaldehyde-fixed erythrocytes at > or = 100 ng/ml and was inhibited by lactose and EDTA (> or = 2 mM) and high concentrations (> 100 mM) of glucose and sucrose, but not by N-acetylglucosamine. BjcuL had no direct hemolytic activity and was devoid of esterase, PLA2 and proteolytic activities. The lectin (up to 200 microg/ml) did not aggregate human platelet-rich plasma (PRP) or washed platelets (WP), nor did it alter the aggregation induced by ADP in PRP or by thrombin in WP. When injected into mouse hind paws, BjcuL (10-100 microg/paw) caused edema and increased vascular permeability, with a maximum effect after 1h that persisted for up to 6 h (edema) or gradually decreased after the peak interval (vascular permeability). No hemorrhage was observed in BjcuL injected paws. In anesthetized rats, B. jararacussu venom (200 microg/kg, i.v.) produced sustained hypotension (maximum decrease of approximately 60%) whereas a similar dose of BjcuL decreased the blood pressure by approximately 15%, with a rapid return to the resting level. PMID- 16309724 TI - Glial inflammation and neurodegeneration induced by candoxin, a novel neurotoxin from Bungarus candidus venom: global gene expression analysis using microarray. AB - Candoxin (PDB #1JGK), a three-finger neurotoxin from Bungarus candidus venom, inhibits post-synaptic neuromuscular and neuronal alpha7nACh-receptors, and induces delayed cell-death throughout the glial population. When applied to cultured human glial cell lines, candoxin (CDX) induced cell death in a concentration (EC(50) approximately 1muM) and time dependent manner. Results of TUNEL-histochemistry further confirm CDX-induced brain (hippocampus, frontal cortex, and temporal regions) damage when administered intracerebroventricularly (i.c.v) in adult mice. In this study, we explored differential gene expression profiles following exposure of human glial (Hs 683) cell lines to CDX at various time intervals using Affymetrix-GeneChips. By means of MAS and GeneSpring analyses, 105 genes whose expression was significantly (P<0.01) altered by at least 3-fold were selected. Results of the genome analysis reveal that the potential role of CDX at molecular level involves the regulation of genes in signal transduction, ubiquitin-inflammation, mitochondrial-dysfunction, and damage-response pathways. In addition, using QRT-PCR and rationally designed specific CDX-binding peptide (P-NT.II), we identified the genes-IL7R, IL13RA2, IL 1beta, TNFRSF12A, GADD45A, CD44 and IFI44-that might play an important role in CDX-induced glial inflammation, DNA-damage and degeneration. These findings reveal new insight into the molecular mechanisms of glial-driven neurodegeneration after exposure to neurotoxins. PMID- 16309725 TI - The infectivity and host range of the ecotropic porcine endogenous retrovirus, PERV-C, is modulated by residues in the C-terminal region of its surface envelope protein. AB - Endogenous retroviral genetic material serves as a reservoir for the generation of retroviral pathogens by recombination between activated endogenous or exogenous infectious agents. Some porcine tissues actively express infectious porcine endogenous retroviruses (PERVs). Of the three classes of PERV characterized to date, two, PERV-A and B, are capable of infecting human cells in vitro, whereas PERV-C cannot. Here, we demonstrate that the PERV-C envelope surface protein (SU) when disassociated from its C-terminus binds human cells. Further, we show that PERV-C binding to human cells is not inhibited in 293 cells productively infected with PERV-A, confirming that the molecule PERV-C interacts with on human cells is distinct from that used by PERV-A. Moreover, we demonstrate that the envelope region encompassing the proline-rich region is required for binding to cells in addition to the putative variable region A (VRA) and B (VRB). The region in the C-terminus of the SU that alters the binding and infectivity properties of PERV-C differs by only nine residues from the analogous region of PERV-A. Caution may be warranted even when a xenotransplantation product is from source pigs that do not express human-tropic viruses, as minimal mutations within PERV-C combined with selection in a human recipient could render PERV-C infectious in humans. PMID- 16309726 TI - HIV-1 tropism for the central nervous system: Brain-derived envelope glycoproteins with lower CD4 dependence and reduced sensitivity to a fusion inhibitor. AB - We previously described envelope glycoproteins of an HIV-1 isolate adapted in vitro for growth in microglia that acquired a highly fusogenic phenotype and lower CD4 dependence, as well as resistance to inhibition by anti-CD4 antibodies. Here, we investigated whether similar phenotypic changes are present in vivo. Envelope clones from the brain and spleen of an HIV-1-infected individual with neurological disease were amplified, cloned, and sequenced. Phylogenetic analysis demonstrated clustering of sequences according to the tissue of origin, as expected. Functional clones were then used in cell-to-cell fusion assays to test for CD4 and co-receptor utilization and for sensitivity to various antibodies and inhibitors. Both brain- and spleen-derived envelope clones mediated fusion in cells expressing both CD4 and CCR5 and brain envelopes also used CCR3 as co receptor. We found that the brain envelopes had a lower CD4 dependence, since they efficiently mediated fusion in the presence of low levels of CD4 on the target cell membrane, and they were significantly more resistant to blocking by anti-CD4 antibodies than the spleen-derived envelopes. In contrast, we observed no difference in sensitivity to the CCR5 antagonist TAK-779. However, brain derived envelopes were significantly more resistant than those from spleen to the fusion inhibitor T-1249 and concurrently showed slightly greater fusogenicity. Our results suggest an increased affinity for CD4 of brain-derived envelopes that may have originated from in vivo adaptation to replication in microglial cells. Interestingly, we note the presence of envelopes more resistant to a fusion inhibitor in the brain of an untreated, HIV-1-infected individual. PMID- 16309727 TI - The role of saccades in exerting voluntary control in perceptual and binocular rivalry. AB - We have investigated the role of saccades and fixation positions in two perceptual rivalry paradigms (slant rivalry and Necker cube) and in two binocular rivalry paradigms (grating and house-face rivalry), and we compared results obtained from two different voluntary control conditions (natural viewing and hold percept). We found that for binocular rivalry, rather than for perceptual rivalry, there is a marked positive temporal correlation between saccades and perceptual flips at about the moment of the flip. Across different voluntary control conditions the pattern of temporal correlation did not change (although the amount of correlation did frequently, but not always, change), indicating that subjects do not use different temporal eye movement schemes to exert voluntary control. Analysis of the fixation positions at about the moment of the flips indicates that the fixation position by itself does not determine the percept but that subjects prefer to fixate at different positions when asked to hold either of the different percepts. PMID- 16309728 TI - Specifying the distractor inhibition account of attention-induced motion blindness. AB - There is growing evidence that motion perception is modulated by visual selective attention. In the 'attention-induced motion blindness' paradigm the detection of coherent motion in a random dot kinematogram (RDK) is impaired in a rapid serial presentation task [Sahraie, A., Milders, M., & Niedeggen, M. (2001). Attention induced motion blindness. Vision Research, 41, 1613-1617]. The effect depends on irrelevant motion episodes (distractors) prior to the target. In this study, we show that both the number and timing of distractors affect detection performance, allowing for implications on the build-up and release of inhibition. Furthermore, we rule out the possibility that subjects falsely classify targets as distractors due to uncertainty of temporal order. PMID- 16309729 TI - The erosion behaviour of biologically active sewer sediment deposits: observations from a laboratory study. AB - The erosion behaviour of various fine-grained sediment deposits has been investigated in laboratory experiments. This work mainly focused on tests using sewer sediment in which strong biochemical reactions were observed during the deposit formation period. A small number of initial tests were conducted in which the deposits were made from mixtures of "clean" mineral and organic sediments. The erosion behaviour observed in these tests was compared with the erosion characteristics for sediments taken from deposits in a sewer. The impact of the biological processes on physical properties such as bulk density, water content, deposit structure and the erosive behaviour as a function of bed shear stress are quantified and discussed. Based on these observations it is believed that bio processes weaken the strength of the in-pipe sediment deposits. A significantly weaker sediment surface layer was observed during deposition under quiescent oxygen-rich conditions. This resulted in a deposit with low shear strength which may be a cause of a first foul flush of suspended sediment when flow rates were increased. Comparison between tests with sewer sediments and the artificial representative surrogates suggested that the deposits of the later did not correctly simulate the depositional development and the resultant erosion patterns observed with the more bio-active sewer sediment. PMID- 16309730 TI - A survey of polycyclic musks in selected household commodities from the United States. AB - Occurrence of the polycyclic musks, 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8 hexamethylcyclopenta[g]-2-benzopyran (HHCB) and 7-acetyl-1,1,3,4,4,6-hexamethyl 1,2,3,4-tetrahydronapthalene (AHTN), in wastewater influent and effluent, as well as in surface waters, has been reported. HHCB and AHTN were also reported to occur in human and wildlife tissues. The major sources for HHCB and AHTN to wastewater are thought to be consumer products such as shampoos, deodorants, laundry detergents, and household surface cleaners. However, the levels of HHCB and AHTN in consumer products are not known. For evaluation of the sources of human and environmental exposures, characterization of levels of HHCB and AHTN in consumer products is needed. In this study, we measured concentrations of HHCB (Galaxolide), AHTN (Tonalide), and HHCB-lactone (Galaxolidone) from a variety of consumer products, including perfumes, body lotions, and deodorants. Concentrations of HHCB, AHTN, and HHCB-lactone in consumer products ranged from <5 ng/g to over 4000 microg/g, <5 ng/g to 451 microg/g, and <5 ng/g to 217 microg/g, respectively. The highest concentrations were found in perfumes, body creams and lotions, and deodorants. The results suggest that a wide variety of source materials exist for HHCB and AHTN, and that these materials are used on a daily basis. PMID- 16309731 TI - Microbial and kinetic characterization of pure and mixed cultures aerobically degrading 4-nitrophenol. AB - The molecular and kinetic characterization of a microorganism able to aerobically degrade 4-nitrophenol (4NP) is presented. The microorganism was isolated from a mixed culture operating in a laboratory-scale sequencing batch reactor with an aerobic anoxic cycle. It was identified as a member of Ralstonia genus within Betaproteobacteria. It is a gram negative coccobacillum (cell length of 2-3 microm) able to aerobically store lipid inclusions when grown aerobically on nitrophenol as the sole carbon source in the range of tested concentrations (80 320 mg l(-1)). Batch kinetic tests were performed with the pure culture, while the kinetics of the mixed biomass was directly investigated in the reactor. For pure cultures exponential growth was observed, with growth rate values in the range of 2-6 d(-1); in experiments with the mixed cultures 4NP concentrations were correlated with growth using the Haldane equation (k(max) = 0.30 mg 4NP mg( 1) VSSh(-1); K(s) = 55 mg 4NPl(-1) and K(I) = 15 mg 4NPl(-1)). Observed pure culture growth rates were higher than those of mixed cultures. This result can be explained by considering that in mixed culture the biomass is evaluated as volatile suspended solids, including both specialized biomass for 4NP removal and denitrifying bacteria. PMID- 16309732 TI - Early hominid dental remains from Members 4 and 5 of the Sterkfontein Formation (1966-1996 excavations): catalogue, individual associations, morphological descriptions and initial metrical analysis. AB - The fossils recovered from the Sterkfontein Formation represent, without doubt, the largest collection of early hominid specimens from a single locality. Among the over 600 entries in the catalogue of fossil hominid specimens recovered since 1966, there are 242 dental remains (isolated teeth, jaws with two or more teeth, isolated teeth in association) for a total number of 495 teeth. The aim of this paper is to provide morphological descriptions of all hominid dental specimens recovered between 1968 and 1996 from areas presently known as Members 4 and 5 of the Sterkfontein site. Together with the descriptions, explanatory catalogue information is provided, along with basic measurements and summary statistics. This paper consists of six sections, with descriptive tables: (1) Catalogue of dental remains, arranged numerically. This includes isolated teeth, specimens with teeth still in position within their jaws, and specimens comprised of isolated teeth in association. (2) List of specimens with more complete dentition and the numbers of available teeth per tooth class. (3) List of specimens subdivided in tooth class, with an indication of their preservation, of the wear, if any, and with measurements (mesiodistal and buccolingual diameters) of the individual teeth. (4) List of associations of isolated teeth as individuals i.e. dental remains that can be associated with one another. Some remarks on the relative abundance of maxillary versus mandibular teeth, and on the numbers of available teeth are presented. (5) Morphological descriptions. (6) Summary statistics for the entire Sterkfontein sample (thus including specimens recovered both before and after 1966) and updated descriptive statistics for South African early hominids (A. africanus, A. robustus, South African early Homo). We have compared the coefficients of variation for the MD and BL diameters of the permanent teeth of the Sterkfontein Member 4 hypodigm of A. africanus with the hypodigms of the early hominid taxa. The results show that the Sterkfontein Member 4 sample is not consistently more variable than the other fossil hominid samples analysed; it turned out to be generally less variable than H. habilis sensu lato and A. boisei; it shows overall similar levels of variability to A. afarensis and higher levels than A. robustus. These results, per se, do not provide evidence of the existence of multiple species in the Sterkfontein Member 4 sample. PMID- 16309733 TI - Adenosine A(3) receptor mediated coronary vasodilation in the rat heart: changes that occur with maturation. AB - Adenosine A(2B) and A(3) receptors (ADOR) have been reported to induce coronary vasodilation in the rat. This study investigated the effect of age on ADORA(3) mediated coronary responses using hearts from rats aged 6-8 weeks (immature), 16 18 weeks (young) and 52-54 weeks (mature) perfused in Langendorff mode. APNEA (ADORA(3)>ADORA(1) agonist) was observed to activate at least two receptor subtypes to mediate a biphasic vasodilator response in hearts from immature rats. The potency of APNEA at the high affinity site was enhanced by alloxazine (ADORA(2B) antagonist) and reduced when combined with MRS1191 (ADORA(3) antagonist). This indicates that the high affinity phase is the ADORA(3), and ADORA(2B) signalling is likely to play a negative regulatory role towards the ADORA(3) mediated response. The activity at this site was also reduced with maturation. The low affinity site was inhibited by alloxazine but not MRS1191, indicating that this response is mediated by the ADORA(2B) or another receptor subtype. The response at this site did not alter with age. Cl-IB-MECA (ADORA(3) agonist) produced monophasic responses that were inhibited by alloxazine but remained unaffected by MRS1191 in all age groups. In addition the potency of Cl IB-MECA does not change in hearts from PTX-treated rats. However, the maximal responses increased, indicating G(i) protein independent and dependent signalling. Q-PCR analysis of rat hearts indicated the presence of an ADORA(3) splice variant (ADORA(3i)), which increased in mRNA expression with age. Cl-IB MECA responses may be mediated by this ADORA(3i). In conclusion, APNEA mediates coronary vasodilation in the rat heart via at least two receptor sites, the ADORA(3) and ADORA(2B). ADORA(3) responses are reduced while ADORA(2B) remain unchanged with maturation. In addition, the splice variant ADORA(3i) may contribute to coronary responses in the rat heart. PMID- 16309734 TI - Metals in soils of children's urban environments in the small northern European city of Uppsala. AB - Metals occur naturally in soil, but contents are generally increased in the urban environment due to anthropogenic activities. The presence of elevated metals in soils of the urban environment has been recognized as an important source of metal intake in children and is linked to elevated metal levels in children's blood. Several metals have undesirable health effects, especially on children due to their still developing nervous system and small body volumes. Playgrounds are where urban children spend most of their time outdoors and are also where children most frequently come in contact with soil. Elevated contents of metals in playgrounds are therefore of great concern for children's wellbeing. This study investigates the soil metal content of 25 playgrounds located in different land use areas in urban Uppsala, Sweden's fourth largest city. Uppsala covers an area of approximately 100 km2 and has a population of 136,000. The soil samples were analysed for 12 metals (Al, As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, W, Zn) using aqua regia. Median metal contents were found to be 1.8, 3.4, 0.21, 32, 25, 2.5, 0.14, 494, 19, 26, 0.35 and 84 mg kg(-1) soil for each of the above metals, respectively. The median clay content was around 20% while the organic matter content was measured by loss on ignition at a median of 8%. The land use areas included industrial land, the city center, road verges, natural land and former industrial land. The results showed that land use did not have the expected large influence on the total metal contents of the soils tested. The clay content together with the age of the site proved to be a more important factor. Sites with elevated clay contents had in general elevated metal contents, which were explained by the relatively high adsorption capacity of clay particles. The soils at sites where land use had not been altered since the 1800s had increased metal contents compared to playgrounds constructed in the late 1900s. The immobility of metals once they had entered the soil system was the reason for increased metal content in soils of old playgrounds. It was concluded that in cities with few internal pollution sources, the soil characteristics of the site and the time the soil has been on-site to accumulate metal residues become important factors in determining the soil metal content. PMID- 16309735 TI - Metastin and its G protein-coupled receptor, GPR54: critical pathway modulating GnRH secretion. AB - Photoperiod, food availability, temperature, stress, and hormonal cues are some of the varied signals used by mammalian species to activate or suppress their timing of sexual maturation. All ultimately converge upon gonadotropin-releasing hormone (GnRH) secretion from the hypothalamus. Through its stimulation of LH and FSH from the pituitary, GnRH represents a critical step in the reproductive cascade. While few dispute this central role of GnRH, little is understood of the mechanisms influencing the developmental fate and physiologic controls of GnRH neurons. Identification of the signals which modulate pulsatile GnRH secretion is critical to advancing understanding of normal puberty and reproductive competency. The recent identification of loss-of-function mutations in GPR54, a receptor for kisspeptin-1, has highlighted a new pathway for the timing of puberty and reproductive control. PMID- 16309736 TI - Pubertal hormones organize the adolescent brain and behavior. AB - Maturation of the reproductive system during puberty results in elevated levels of gonadal steroid hormones. These hormones sculpt neural circuits during adolescence, a time of dramatic rewiring of the nervous system. Here, we review the evidence that steroid-dependent organization of the adolescent brain programs a variety of adult behaviors in animals and humans. Converging lines of evidence indicate that adolescence may be a sensitive period for steroid-dependent brain organization and that variation in the timing of interactions between the hormones of puberty and the adolescent brain leads to individual differences in adult behavior and risk of sex-biased psychopathologies. PMID- 16309737 TI - "Coffee, tea and me": moderate doses of caffeine affect sexual behavior in female rats. AB - The present study evaluated the effects of acute caffeine administration on paced mating behavior and partner preference in ovariectomized rats primed with estrogen and progesterone. In Experiment 1, female rats were tested for paced mating behavior following acute administration of caffeine (15 mg/kg). Caffeine shortened the latency to return to a male following an ejaculation. Although this dose of caffeine did not alter the likelihood of leaving a male after receiving sexual stimulation, locomotor activity did increase significantly. Experiment 2 evaluated the dose response characteristics of caffeine (7.5, 15, 30 mg/kg) administration on paced mating behavior. Replicating Experiment 1, caffeine at the lower doses shortened the latency to return to a male following an ejaculation. Finally, to determine whether the effects of caffeine (15 mg/kg) on contact-return latency reflect a change in sexual motivation or merely an inability to inhibit locomotion, rats were tested for partner preference (intact male vs. estrous female) following caffeine administration (Experiment 3). Although caffeine did not disrupt preference for a sexual partner, caffeine selectively increased visits to the male when physical contact was possible. Collectively, these results suggest that the effects of caffeine on female mating behavior may reflect an increase in both sexual motivation and locomotor activity. PMID- 16309738 TI - Carotenoid actions and their relation to health and disease. AB - Based on extensive epidemiological observation, fruits and vegetables that are a rich source of carotenoids are thought to provide health benefits by decreasing the risk of various diseases, particularly certain cancers and eye diseases. The carotenoids that have been most studied in this regard are beta-carotene, lycopene, lutein and zeaxanthin. In part, the beneficial effects of carotenoids are thought to be due to their role as antioxidants. beta-Carotene may have added benefits due its ability to be converted to vitamin A. Additionally, lutein and zeaxanthin may be protective in eye disease because they absorb damaging blue light that enters the eye. Food sources of these compounds include a variety of fruits and vegetables, although the primary sources of lycopene are tomato and tomato products. Additionally, egg yolk is a highly bioavailable source of lutein and zeaxanthin. These carotenoids are available in supplement form. However, intervention trials with large doses of beta-carotene found an adverse effect on the incidence of lung cancer in smokers and workers exposed to asbestos. Until the efficacy and safety of taking supplements containing these nutrients can be determined, current dietary recommendations of diets high in fruits and vegetables are advised. PMID- 16309739 TI - The influence of sediment resuspension on the degradation of phenanthrene in flow through microcosms. AB - The effect of sediment resuspension on the mineralization of phenanthrene was examined in microcosms and sediment slurries. In computer-controlled, flow through microcosms, 14C-phenanthrene-amended sediments were resuspended into overlying oxic water at frequencies of 12, 4, 1, 0.25 and 0 d(-1). In slurry bottle experiments 14C-phenanthrene-amended sediments were continuously resuspended under oxic (excess air headspace) and anoxic (N2 headspace) conditions and mineralization was measured at periods from 2 h to 7 days. Our main findings were: (1) mineralization rate constants from the microcosms ranged from 0.001 to 0.01 d(-1) and increased with frequency of resuspension, (2) these rates fell between those measured in oxic and anoxic slurries and were predicted within a factor of 2.5 by a model in which mineralization depended on the degree of oxygen exposure, and (3) the phenanthrene-degrading bacterial community was more active in resuspended sediments incubated in the microcosms than in sediments which were not resuspended, or which were stored under refrigeration. We conclude from these experiments that the effects of sediment resuspension on phenanthrene degradation are consistent with a primary role of average oxygen exposure, and also an alteration in the PAH-degrading activity of microbial populations. PMID- 16309740 TI - Constitutive models for impacted morsellised cortico-cancellous bone. AB - Constitutive models are developed, based on the results of confined compression testing, to describe the visco-elastic, and non-linear elasto-plastic behaviour of morsellised cortico-cancellous bone (MCB). It is found that the elastic modulus, E of MCB can be expressed as a linear function of the applied pressure, p. E varied from 3 to 30N/mm(2) for pressures up to 1N/mm(2). The visco-elastic behaviour of MCB can be described using a fourth-order Prony series. The plastic behaviour of MCB can be described using a Drucker Prager Cap (DPC) yield criterion, in which consolidation behaviour is described using an exponential function. The developed relationships allow MCB to be included in a realistic manner in finite element models, for example of the acetabular construct, following revision hip arthroplasty, carried out using the Slooff-Ling impaction grafting technique. PMID- 16309741 TI - Formation of hydroxyapatite onto glasses of the CaO-MgO-SiO2 system with B2O3, Na2O, CaF2 and P2O5 additives. AB - New bioactive glasses with compositions based on the CaO-MgO-SiO(2) system and additives of B(2)O(3), P(2)O(5), Na(2)O, and CaF(2) were prepared. The in vitro mineralization behaviour was tested by immersion of powders or bulk glasses in simulated body fluid (SBF). Monitoring of ionic concentrations in SBF and scanning electron microscopy (SEM) observations at the surface of the glasses were conducted over immersion time. Raman and infrared (IR) spectroscopy shed light on the structural evolution occurring at the surface of the glasses that leads to formation of hydroxyapatite. PMID- 16309742 TI - Invocation of coercion context in compliance communication -- power dynamics in psychiatric care. AB - This article draws on observations from ethnographic fieldwork to develop a theoretical understanding of the power dynamics in psychiatric care. The aim is to analyze how psychiatric clinicians solve compliance problems by invoking "coercion context". It is suggested that clinicians take a rather instrumental approach to laws regulating coercive intervention. Clinicians may invoke a coercion context even with voluntary patients. For example, they may use wordings that connote coercion, or they may make use of how treatment wards are set up to accommodate involuntary patients, thus stalling voluntary patients who cannot exit through locked doors. A coercion context can also be invoked to solve mundane practical problems, e.g. when clinicians talk about "coerced showers". The management of information and maintaining a suitable "awareness context" with regards to coercion is an essential feature in clinical attempts to achieve compliance from patients. In conclusion, the notion of coercion context helps explain the confusing findings from previous research about patients' apparent misconceptions of their formal legal status. Furthermore, it is argued that research that rely on decontextualised, objectifications of "coercion" risk to miss the meaning coercion is assigned in everyday clinical practice. PMID- 16309743 TI - Simple effective surgery for involutional entropion suitable for the general ophthalmologist. AB - OBJECTIVE: To compare the long-term success, recurrence, and complication rate of involutional entropion surgery using the lateral tarsal strip and everting sutures when performed by surgeons in training (resident or fellow) and specialist oculoplastic surgeons (attending supervising physician). DESIGN: Prospective, interventional, comparative, clinical case series. PARTICIPANTS: Adult patients with involutional entropion. METHODS: Lateral tarsal strip and everting sutures (LTS+ES) by residents, fellows, or attending supervising physician. A minimum of 12 months of postoperative follow-up was required. MAIN OUTCOME MEASURES: Patients' symptoms and clinical examination to confirm a normal eyelid position (no entropion or secondary ectropion) at rest and with forced orbicularis contraction with the topical amethocaine (tetracaine) test. This test is described. RESULTS: Fifty-five consecutive patients, aged 57 to 91 years (mean, 77 years) underwent LTS+ES surgery on 62 eyelids. Surgery was performed by a consultant ophthalmic oculoplastic surgeon (attending supervising physician) in 8 eyelids and by 20 different trainees, residents, and fellows in 54 eyelids. Six patients died (11%) within 6 months of surgery and 2 patients (3.5%) were lost to follow-up, resulting in 47 evaluable patients (54 eyelids). The follow-up period was 12 to 34 months (mean, 18 months). Fifty-three of 54 eyelids (98%) had a successful outcome with no recurrence. The surgery was effective when performed by different grades of surgeon (P>0.4). CONCLUSIONS: The LTS+ES is a simple operation for the correction of involutional entropion that can be performed effectively by both residents and fellows. PMID- 16309744 TI - Neuronal regeneration partially compensates the delayed neuronal cell death observed in the hippocampal CA1 field of soman-poisoned mice. AB - Soman poisoning induces long-term neuropathology characterized by the presence of damaged neurons up to 2 months after exposure in various central brain areas, especially the hippocampal CA1 layer. Rapid depletion of this layer could therefore be expected. Surprisingly, the CA1 layer remained consistently visible, suggesting delayed death of these damaged neurons, potentially accompanied by neuronal regeneration. To address this issue, mice were exposed to a convulsive dose of soman (110 microg/kg followed by 5.0mg/kg of atropine methyl nitrate (MNA) 1 min later) and brains were collected from day 1 to day 90 post-exposure. Damaged and residual healthy neurons were quantified on brain sections using hemalun-phloxin and fluorojade staining or neuronal nuclei antigen (NeuN) immunohistochemistry. On post-soman day 1, a moderate neuronal cell death was noticed in the hippocampal CA1 layer. In this area, an important and steady quantity of damaged neurons (about 48% of the whole pyramidal neurons) was detected from post-soman day 1 to day 30. Thus, throughout this period, damaged neurons seemed to survive, as confirmed by the unmodified depth of the hippocampal CA1 layer. The dramatic disappearance of the damaged neurons occurred only later during the experiment and was almost complete at day 90 after soman exposure. Interestingly, between day 30 and day 90 following poisoning, an increase in the number of residual healthy pyramidal neurons was observed. These different kinetic patterns related to the density of total, damaged and residual healthy neurons after soman poisoning demonstrate that neuronal regeneration is delayed in the hippocampal CA1 layer and is concomitant to the death of damaged neurons. PMID- 16309745 TI - Diagnosis and evaluation of febrile infants under 4 months of age in Japan by using RT-PCR for enterovirus. AB - OBJECTIVES AND METHODS: In order to establish an effective management for febrile infants under 4 months of age we analysed the causes of fever by using bacterial work up and a highly sensitive RT-PCR method during a 3-year-period. From February 1998 to January 2001, 263 infants under 4 months of age were admitted to our hospital because of fever (>38.0 degrees C) and enrolled in this study. RESULTS: Eighty-nine cases (33.8%) were diagnosed as lower respiratory infections, and 62 (23.5%) as urinary tract infections. Aseptic meningitis was found in 35 cases (13.2%) and fever of unknown origin in 22 (8.3%). Sepsis and purulent meningitis were found in three and two infants, respectively. Aseptic meningitis was the most common disease in infants below 30 days. Aseptic meningitis and fever of unknown origin revealed no seasonable tendency. Twelve cases out of 20 tested infants with aseptic meningitis showed positive results for enterovirus in CSF. CONCLUSION: Clinicians should consider age, sex and seasons when evaluating and treating infants with fever. Previously lumbar puncture, urinalysis and chest X-ray were recommended to avoid useless assays. These results also revealed that RT-PCR is a useful and significant assay to establish an exact diagnosis in very young infants. PMID- 16309746 TI - Bilateral alteration in stepping pattern after unilateral motor cortex injury: a new test strategy for analysis of skilled limb movements in neurological mouse models. AB - Mice are becoming increasingly popular to model neurological disease and motor system dysfunction. For evaluation of discrete, chronic motor impairments, skilled limb movements represent a valuable extension of standard mouse test batteries. This study introduces an efficient and sensitive test strategy for comprehensive assessment of skilled fore- and hind-limb stepping in mice. Adult C57BL/6 mice were trained and video-recorded in two walking tasks, the widely used rotorod test and a new ladder rung task. The animals then received a unilateral ischemic lesion in the motor cortex forelimb and hind limb area and were video-recorded on days 12 and 26 post-lesion. Forelimb and hind limb stepping movements were rated using a combination of endpoint measures and qualitative assessment. The results showed that while animals maintained a weight supported gait, posture and stepping movements were abnormal at both post operative intervals. The rotorod analysis revealed stepping deficits in both forelimbs that led to adoption of compensatory movement strategies. The ladder rung task revealed stepping errors in ipsi- and contralateral fore- and hind limbs. The findings demonstrate that this test strategy provides comprehensive assessment of motor impairments in mice and that qualitative movement analysis is a valuable tool for elaborating subtle motor disturbances. PMID- 16309747 TI - Lymphocyte phenotyping and NK cell activity analysis in pregnant NOD/SCID mice. AB - The purpose of this study was to outline the fertility features of non-obese diabetic (NOD)/LtSz-scid/scid (NOD/SCID [severe combined immunodeficiency] for short) mice, and to evaluate the effects of NK cell subsets on the pregnancy outcomes of the syngeneic NOD/SCID x NOD/SCID mating combination. Firstly, lymphocyte phenotyping was performed with flow cytometry to confirm the multiple immunodeficits in NOD/SCID mice. Fertility features were assessed in NOD/SCID x NOD/SCID mice and compared with non-immunodeficiency control BALB/c x BALB/c mice. Although the presence of NK cell deficit is apparent in NOD/SCID mice, a certain level of remnant NK activity could be observed in these mice. The remnant NK cell activity was stimulated with polyinosinic-polycytidylic acid (polyIC), or inhibited with anti-asialo GM1 (ASGM1) anti-serum, respectively. The effects of these factors on pregnancy outcomes were evaluated after administration. Roughly normal fertility could be observed in NOD/SCID x NOD/SCID mice. However, a slightly larger sized litter was observed in polyIC-treated NOD/SCID x NOD/SCID mice compared with control NOD/SCID mice. In contrast, embryo resorption was boosted after ASGM1 injection, and correlated subsequently with a smaller litter size. It indicates that the remnant NK cell activity in NOD/SCID mice may be beneficial to feto-maternal tolerance during pregnancy. PMID- 16309748 TI - Velopharyngeal function after two different types of pharyngoplasty. AB - OBJECTIVE: To evaluate velopharyngeal function after two different types of pharyngoplasty: pharyngeal flap (PF) and sphincter pharyngoplasty (SP). DESIGN: Two groups of patients operated on with pharyngeal flaps or sphincter pharyngoplasty were studied prospectively. SETTING: The study was carried out at the cleft palate clinic of the Hospital Gea Gonzalez in Mexico City. PATIENTS: Fifty-eight cleft palate patients were studied. All patients had undergone palatal repair and showed residual velopharyngeal insufficiency. Twenty-five patients were operated on using a pharyngeal flap. Twenty-three patients were operated on using a sphincter pharyngoplasty. MAIN OUTCOME MEASURES: Pharyngeal muscle function was evaluated using selective electromyography (EMG) and simultaneous videonasopharyngoscopy (VNP). The superior constrictor muscle was approached directly. The levator veli palatini was included in the central pharyngeal flap. The palatopharyngeus were included in the lateral and superiorly based surgical flaps inserted on the posterior pharyngeal wall. All patients showed a complete velopharyngeal closure after surgery. RESULTS: None of the patients showed muscle activity on the central pharyngeal flaps. None of the patients showed muscle activity on the lateral flaps of the sphincter pharyngoplasties. All the patients showed strong muscle activity of the superior constrictor. CONCLUSIONS: It is concluded that lateral pharyngeal flaps in cases of sphincter pharyngoplasty and the central pharyngeal flap in cases of pharyngeal flap, show absence of intrinsic activity during speech. The participation of these structures when velopharyngeal closure occurs during speech is rather passive. These flaps, central or lateral, increase tissue volume on specific areas; their movements are produced by the underlying superior constrictor. PMID- 16309749 TI - Chronic otitis media with effusion and Helicobacter pylori. AB - OBJECTIVE: The aim of this study is to investigate the presence of Helicobacter pylori (HP) in the middle ear effusion by Campylobacter-like organism (CLO) test and whether it has a role in the ethiopathogenesis of chronic otitis media with effusion (OME). STUDY DESIGN: A prospective randomized and controlled study. METHODS: This study was performed with 45 patients with the diagnosis of chronic OME and adenoid hypertrophy, between the ages 3 and 13 (median 6). Thirty patients constituted the study group (18 male (60%) and 12 (40%) female). Adenoidectomy with myringotomy with ventilation tube insertion were performed to this group. Middle ear effusion and adenoid tissue pieces were collected and H. pylori presence was investigated by "CLO" testing. Fifteen patients of the matching age group (9 male (66.7%) and 6 (33.3%) female) constituted the control group to whom adenoidectomy with myringotomy were performed but no middle ear effusion could be determined (empty myringotomy patients). The wash out liquid of middle ear and pieces of adenoid tissue samples were also collected from the control group. By using CLO testing, the presence of H. pylori was investigated in the adenoid tissues and middle ear of the empty myringotomy patients. RESULTS: In 20 (66.6%) patients of the study group, CLO testing was positive in the middle ear effusions. None of the patients demonstrated positive CLO test in the wash out liquid of middle ear. There was significant difference of positive CLO testing in the middle ear effusions of two groups (p<0.001). CONCLUSIONS: These findings showed us that presence of HP in the middle ear effusion using CLO testing and this may be responsible for the ethiopathogenesis of chronic OME. PMID- 16309750 TI - Acute otitis media and mastoiditis following cochlear implantation. AB - OBJECTIVE: To examine the incidence of acute otitis media (AOM) and mastoiditis in children after cochlear implantation (CI) and to evaluate the role of mastoidectomy in decreasing the rate of AOM in implanted children by comparing two surgical techniques: the Posterior tympanotomy approach (MPTA, with mastoidectomy) and the Suprameatal approach (SMA, without mastoidectomy). METHODS: A retrospective study was conducted on 234 children up to 16 years of age who underwent CI between 1993 and 2003 in our department. The children were divided into two groups according to the surgical technique that had been used for the implantation: the MPTA group and the SMA group. RESULTS: Part of the children with a history of pre-implantation AOM (22 of 29 in MPTA group and 26 of 38 in SMA group) did not suffer from AOM post-CI (p=0.59), and an incidence of AOM after CI in children who did not have history of AOM prior to implantation (13 patients of MPTA group and 15 patients of SMA group) was unrelated to surgical approach (p=0.65). The incidence of pre-implantation AOM was similar for the two groups and declined after CI unrelated to performing of mastoidectomy in surgical technique. Overall, 47 children (20.1%) had post-CI AOM compared to 67 children (28.6%) who had pre-CI AOM. Mastoiditis developed in 11 children (4.7%), all 11 in the MPTA group. A subperiosteal abscess was incised and drained with the retroauricular approach in three of these children and the others were managed with intravenously administered ceftriaxone 50mg/kg/day for 3-5 consecutive days, followed by a course of oral cephalexin until there is complete clinical resolution of the effusion in the middle ear. The implants were preserved in all cases. Seven out of 11 children with mastoiditis had no history of AOM prior to implantation. CONCLUSIONS: AOM and mastoiditis represent common complications of CI that can be successfully treated with the prompt use of antibiotics. However, the subperiosteal abscess could require surgical drainage. In our opinion, the decrease of incidence of AOM in implanted children is the result of natural history of otitis media and is unrelated to the surgical approach. PMID- 16309751 TI - Detection of equine herpesvirus type 1 by real time PCR. AB - A real-time PCR assay was developed for detection and quantitation of equid herpesvirus type 1 (EHV-1). The sensitivity of the assay was compared with an established nested-PCR (n-PCR). The real-time PCR detected 1 copy of target DNA, with a sensitivity 1 log higher than gel-based n-PCR. The assay was able to detect specifically EHV-1 DNA in equine tissue samples and there was no cross amplification of other horse herpesviruses. Real-time PCR was applied to determine EHV-1 load in tissue samples from equine aborted fetuses. The high sensitivity and reproducibility of the EHV-1-specific fluorogenic PCR assay, combined with the wide dynamic range and the high throughput, make this method suitable for diagnostic and research applications. PMID- 16309752 TI - Development of a real time RT-PCR to detect and type ovine pestiviruses. AB - A real time one-step RT-PCR was designed to detect and type border disease virus (BDV), bovine viral diarrhea virus (BVDV) type 1 and BVDV type 2 in ovine samples. The real time RT-PCR was shown to behave in a linear manner and had limits of detection of 100-1000 copies of viral RNA as judged by in vitro transcribed RNA. The real time RT-PCR was validated on 50 clinical samples from UK flocks and was more sensitive than a virus isolation and a classical nested RT PCR (nRT-PCR). The results of real time RT-PCR virus typing agreed completely with sequencing. The majority of ovine isolates were BDV; a small proportion were BVDV type 1. BVDV type 2 was not detected in any sample. This test appears reliable and can be used for the typing of ovine pestiviruses in the UK. PMID- 16309753 TI - Circuits that build visual cortical receptive fields. AB - Neural sensitivity to basic elements of the visual scene changes dramatically as information is handed from the thalamus to the primary visual cortex in cats. Famously, thalamic neurons are insensitive to stimulus orientation whereas their cortical targets easily resolve small changes in stimulus angle. There are two main types of cells in the visual cortex, simple and complex, defined by the structure of their receptive fields. Simple cells are thought to lay the groundwork for orientation selectivity. This review focuses on approaches that combine anatomy with physiology at the intracellular level, to explore the circuits that build simple receptive fields and that help to maintain neural sensitivity to stimulus features even when luminance contrast changes. PMID- 16309754 TI - Antiviral effect of the heparan sulfate mimetic, PI-88, against dengue and encephalitic flaviviruses. AB - Many viruses, including flaviviruses, display affinity for cell surface heparan sulfate (HS) proteoglycans with biological relevance in virus attachment/entry. This raises the possibility of the application of HS mimetics in antiviral therapy. We have evaluated the antiviral effect of the sulfated polysaccharides, suramin, pentosan polysulfate (PPS) and PI-88, which are currently approved or in trial for clinical use, against dengue virus (DEN) and the encephalitic flaviviruses, Japanese encephalitis virus, West Nile virus, and Murray Valley encephalitis virus. A flow cytometry-based method for the measurement of inhibition of virus infectivity was developed, which showed the in vitro antiviral activity of the three compounds, albeit with differences in efficiency which were virus-dependent. The 50% effective concentration (EC(50)) values for DEN inhibition were in the order: PPS or =126 ms and RMS 20 < or =3.1 muV and could predict atrial fibrillation with a specificity of 77%, a sensitivity of 72%, a positive value of 75%, a negative predictive value of 75% and an accuracy of 75%. Patients with FPD > or =126 ms and RMS 20 < or =3.1 muV had nearly a 7-fold increase risk for recurrence of atrial fibrillation. CONCLUSION: The recurrence of atrial fibrillation after cardioversion could be detected by P wave signal averaged ECG and an analysis of CHRS. These methods seem to be appropriate to show a delayed atrial conduction and a neurovegetative imbalance which might be possible trigger mechanisms of reinitiating of atrial fibrillation. PMID- 16309760 TI - Abnormal regional left ventricular mechanics in treated hypertensive patients with 'normal left ventricular function'. AB - Global systolic and diastolic LV function assessed by conventional echocardiographic indices is often normal in patients with controlled hypertension, with or without left ventricular hypertrophy. However, it is not certain whether regional myocardial function in these patients remains normal. We investigated 26 patients and 10 age matched normal controls, by means of long axis M-mode echocardiography. There was no significant difference in age, sex distribution, heart rate, blood pressure and routine ECG measurements between the two groups. Although there was significant LVH in patients compared to normal controls, LV cavity size and global systolic function, assessed by shortening fraction, ejection fraction and mean velocity of circumferential fibre shortening did not differ between the two groups, nor did LV diastolic function, assessed by the mitral flow pattern. However, LV regional mechanics, as assessed by multiple long axis M-mode echocardiograms differed significantly, in both systole and diastole, between the two groups. Compared to controls, the total longitudinal systolic excursion in both LV free wall and ventricular septum were significantly reduced in patients, and so was maximum early relaxation and atrial contraction in the LV free wall. The mean rate of systolic excursion in all 3 sites did not differ between the two groups, but the mean rate of early relaxation in both LV free wall and ventricular septum was significantly decreased in patients compared to normal controls. In conclusion, the evaluation of LV dysfunction in patients who have achieved good blood pressure control requires more than a conventional echocardiographic assessment. The assessment of regional mechanics described in the present paper offers an easy and sensitive method for the detection of subtle signs of LV mechanical inefficiency associated with LVH. PMID- 16309761 TI - Inflammatory and vasoactive factors in the aspirate from the culprit coronary artery of patients with acute myocardial infarction. AB - BACKGROUND: Besides distal embolization of thrombus and plaque debris, locally increased inflammatory markers at the site of ruptured plaque in acute myocardial infarction (AMI) are thought to have an adverse impact on myocardial reperfusion during primary percutaneous coronary intervention (PCI). However, there is lack of data on such factors. Therefore, we investigated the presence of locally increased inflammatory and vasoactive factors in culprit coronary artery. METHODS: We performed primary PCI with PercuSurge GuideWire system in 18 AMI patients. We collected blood samples from the femoral artery before PCI and from culprit coronary artery after first predilation while inflating the distal protection balloon and after completing PCI. We determined concentrations of C reactive protein, soluble CD40 ligand, Interleukin (IL-6), serotonin, tissue factor, and factor VIIa. RESULTS: While the concentrations of soluble CD40 ligand (2.84+/-3.74 vs 0.98+/-0.63 ng/mL, p=0.004), IL-6 (33.67+/-32.63 vs 17.08+/-21.41 pg/mL, p<0.001), serotonin (2.05+/-0.76 vs 0.92+/-0.60 ng/mL, p<0.001), tissue factor (257.17+/-84.34 vs 154.60+/-87.99 pg/mL, p<0.001) and factor VIIa (34.30+/ 27.30 vs 24.19+/-28.00 ng/mL, p=0.016) were significantly higher in the culprit coronary artery than in the femoral artery, CRP levels did not differ. The locally elevated concentrations of various factors were successfully reduced after multiple aspirations of blood using the PercuSurge GuideWire system. CONCLUSIONS: We found increased levels of soluble CD40 ligand, IL-6, serotonin, tissue factor and factor VII in the culprit coronary artery compared to those in peripheral blood. The clinical impact of such locally increased soluble factors in the culprit coronary artery needs to be investigated in further studies. PMID- 16309762 TI - QT dispersion and left ventricular function after stent placement in acute myocardial infarction. AB - BACKGROUND: QT dispersion is increased in acute myocardial infarction (AMI), but the relation of QT dispersion to left ventricular (LV) function has not yet been fully elucidated. The purpose of this study was to evaluate the relationship between QT dispersion and LV function in patients with successful coronary stenting in AMI. METHODS: Seventy five patients with AMI who underwent percutaneous transluminal coronary angioplasty (PTCA) were enrolled in this study. Corrected QT dispersion was measured before, immediately after, 24 h after, 48 h after, and 6 months after PTCA. Left ventricular ejection fraction (LVEF) was evaluated by left ventriculography at 6 months after PTCA. RESULTS: Corrected QT dispersion at 24 h after and 48 h after PTCA were significantly related to LVEF by univariate analysis (r=-0.282, p<0.05 and r=-0.326, p<0.01, respectively). In multiple regression model, corrected QT dispersion at 24 h after and 48 h after PTCA revealed significant associations with LVEF (R(2)=0.441, coefficient=-0.283, p=0.006 and R(2)=0.411, coefficient=-0.225, p=0.039, respectively), but corrected QT dispersion before, immediately after, and 6 months after PTCA were not associated with LVEF. CONCLUSIONS: Corrected QT dispersion at 24 h after and 48 h after PTCA in AMI correlate with LVEF at 6 months after PTCA. PMID- 16309763 TI - Stenting of a restrictive foramen ovale in a patient with hypoplastic left heart syndrome. AB - Neonates with a severely restrictive foramen ovale and hypoplastic left heart syndrome (HLHS) present as an emergency. We report the case of a newborn in whom left atrial decompression was successfully achieved by stent implantation into the foramen ovale on the first day of life. PMID- 16309764 TI - Obstructive sleep apnea in atrial fibrillation patients. AB - A high prevalence of obstructive sleep apnea has been demonstrated in patients with atrial fibrillation. Our comments want to emphasize the importance of identifying and treating a large proportion of patients with atrial fibrillation who have undiagnosed obstructive sleep apnea as an additional preventive strategy for atrial fibrillation patients. PMID- 16309765 TI - Role of erythropoietin in anemia after heart transplantation. AB - BACKGROUND: Anemia after heart transplantation is common; however, there are scant data on etiology and treatment. This study evaluates type of anemia and the effects of erythropoietin therapy. METHODS: In 37 anemic heart transplant recipients (31 male/59.1+/-10.3 years/hemoglobin <12.0 g/dl), complete anemia work-up was performed including erythropoietin determination. For three months, 12 anemic patients with renal failure (9 male/64.1+/-13.6 years) were treated with 1-3x4000 IU of epoietin beta/week; treatment endpoints were hemoglobin levels and quality of life as determined by questionnaire. RESULTS: In 31 patients no other cause of anemia than renal insufficiency (mean creatinine 1.9+/ 0.9 mg/dl, mean calculated GFR 50.8+/-21.5 ml/min, no hemodialysis) was found; in 93.5% of these patients with renal insufficiency, measured erythropoietin levels were markedly lower than predicted [Beguin Y, Clemons GK, Pootrakul P, Fillet G. Quantitative assessment of erythropoiesis and functional classification of anemia based on measurements of serum transferrin receptor and erythropoietin. Blood 1993; 81(4):1067-1076.]. There was an inverse correlation of hemoglobin levels with serum creatinine/creatinine clearance and a strong trend for inverse correlation of erythropoietin levels. All 12 patients treated with erythropoietin showed a significant increase in hemoglobin levels after three months returning to pre-treatment values within 3 months of cessation of therapy (before study 10.8+/-1.1 g/dl, end of study 14.1+/-1.7 g/dl, three months after end of study 11.6+/-2.1 g/dl; p<0.005). Quality of life was significantly improved in eight patients (75%). CONCLUSIONS: Anemia after heart transplantation is associated with moderate renal failure and low erythropoietin levels in most patients. Erythropoietin therapy resulted in increased hemoglobin levels in all and improved quality of life in 75% of patients. Erythropoietin may be a superior marker of functional renal impairment after heart transplantation; its therapeutic substitution allows effective anemia management and improves quality of life. PMID- 16309766 TI - Strategic integration of hospitals and physicians. AB - A striking development in the healthcare market place has been the formation of strategic relationships between hospitals and physicians. Hospital-physician integration appears to be a response to rapidly expanding managed care health insurance. We examine whether integration lead to efficiency gains from transaction cost economies thereby allowing providers to offer managed care insurance plans lower prices or whether integration is really a strategy to improve bargaining power and thereby increase prices. We find that integration has little effect on efficiency, but is associated with an increase in prices, especially when the integrated organization is exclusive and occurs in less competitive markets. PMID- 16309767 TI - Life in earth: the impact of GM plants on soil ecology? AB - The impact of changes incurred by agricultural biotechnology has led to concern regarding soil ecosystems and, rightly or wrongly, this has focused on the introduction of genetically modified (GM) crops. Soils are key resources, with essential roles in supporting ecosystems and maintaining environmental quality and productivity. The complexity of soils presents difficulties to their inclusion in the risk assessment process conducted for all GM plants. However, a combined approach, informed by both soil ecology and soil quality perspectives, that considers the impacts of GM crops in the context of conventional agricultural practices can provide a regulatory framework to ensure the protection of soils without being overly restrictive. PMID- 16309768 TI - Zebrafish in functional genomics and aquatic biomedicine. AB - The zebrafish (Danio rerio) has many features that make it an ideal model for the study of developmental biology. It is small and easy to contain, it has transparent embryos, it is easy to breed and its early development is well characterized; these same characteristics have also made it an ideal vertebrate model in the areas of biomedicine and biotechnology. In aquaculture, the need for a well-characterized fish model has been satisfied by the zebrafish owing to the availability of functional genomics and molecular biology data to facilitate studies of growth, reproduction, meat quality and disease biology, with the corresponding development of vaccines and therapies. Zebrafish are also increasingly used in toxicogenomics to analyze the effects of toxins and pollutants in the environment, and for creating biomonitors that emit alarm signals when a toxic compound is detected. As detailed in this review, the zebrafish is a versatile and well-characterized model with applications in many fields of study. PMID- 16309769 TI - Randomized blinded clinical trial of intracoronary brachytherapy with 90Sr/Y beta radiation for the prevention of restenosis after stent implantation in native coronary arteries in diabetic patients. AB - BACKGROUND: We report a double-blind, randomized clinical trial of intracoronary beta-radiation for prevention of restenosis after stent implantation in native coronary de novo lesions in diabetic patients. METHODS: After successful stent implantation in native coronary de novo lesions, 106 lesions in 89 diabetic patients were randomly allocated to treatment with beta-radiation with 18 Gy at 1 mm vessel depth (n = 53) or placebo treatment (n = 53). RESULTS: Angiographic analysis at 9 month follow-up revealed a late lumen loss of 0.7+/-0.9 mm in the radiotherapy group versus 1.2+/-0.8 mm in the control group at the injured segment (P = 0.006), 0.9+/-1.0 versus 1.3+/-0.7 mm at the radiated segment (P = 0.02), and 0.9+/-1.0 versus 1.3+/-0.7 mm at the target segment (P = 0.04) (defined as active source length plus 5mm on proximal and distal sites). Binary restenosis rates were significantly lower in the radiation group in all subsegments (injured segment: 10.9 versus 37.3%, P = 0.003; radiated segment: 21.7 versus 49.0%, P = 0.005; target segment: 23.9 versus 49.0%, P = 0.01). Target lesion revascularization for restenosis was required in nine lesions (17.6%) in the radiotherapy group versus 18 (34.0%) in the placebo group (P = 0.05). Late thrombosis occurred in four radiated patients (after premature discontinuation of antiplatelet therapy in all), resulting in a major adverse clinical event rate of 37.2% in the brachytherapy group versus 38.6% in the placebo group (P = ns). CONCLUSIONS: In diabetic patients with de novo coronary lesions, intracoronary radiation after stent implantation significantly reduced restenosis. However, this clinical benefit was reduced by the frequent occurrence of late thrombosis. PMID- 16309770 TI - Site of relapse after chemotherapy alone for stage I and II Hodgkin's disease. AB - BACKGROUND: Short course chemotherapy followed by radiotherapy is a standard treatment for early Hodgkin's disease. There is yet no consensus regarding the appropriate radiotherapy portal following chemotherapy. A good guide to the adjuvant radiotherapy field is the site of relapse in patients treated with chemotherapy alone. PATIENTS AND METHODS: From 1980 to 1996, 61 patients with stage I and II supradiaphragmatic Hodgkin's disease were treated with chemotherapy alone at the Royal Marsden Hospital. We undertook a retrospective review and failure analysis to define the pattern of recurrence. RESULTS: After a median follow-up of 6.5 years, 24 patients had relapsed giving a 5-year relapse rate of 40%. The 5 and 10-year actuarial survival rates were 94 and 89%, respectively with cause-specific survival being 94% at 5 and 10 years. Two-thirds of the relapses were nodal and supradiaphragmatic. Twenty patients (83%) relapsed in the initially involved sites of disease and this was the sole site of recurrence in 11 (45%) of patients. In retrospect, it appeared that at least 12 recurrences could have been prevented by involved field radiotherapy. Review of detailed imaging data (available in 9 out of 11 patients with recurrences in initial sites of disease) showed that the relapses were always in the initially involved nodes. CONCLUSION: After chemotherapy alone in early stage HD most initial recurrences are nodal. Loco-regional recurrences are in the originally involved nodes. Based on limited data it appears that involved nodal RT is equivalent to involved field radiotherapy and may halve the risk of recurrence. PMID- 16309771 TI - Heat stress, plasma concentrations of adrenaline, noradrenaline, 5 hydroxytryptamine and cortisol, mood state and cognitive performance. AB - The primary aims of this paper were to examine the effect of heat stress on working memory, choice reaction time and mood state, and to investigate the relationship between heat induced changes in plasma concentrations of selected neurotransmitters and hormones, and cognition. Heat stress resulted in a deterioration of performance on a central executive task (random movement generation) but not on verbal and spatial recall, and choice reaction time tasks. Perceptions of vigour decreased and fatigue increased following exposure to heat stress. Plasma concentrations of cortisol and 5-hydroxytryptamine significantly increased following exposure to heat. Regression analyses showed that percent body mass loss and change from baseline (Delta) concentrations of cortisol, post exposure to heat, were significant predictors of Delta random movement generation and Delta fatigue. A secondary purpose was to examine the effect of recovery on cognition and mood. Following recovery, the performance of the central executive task was poorer than pre-treatment. Mood states, catecholamines and 5 hydroxytryptamine concentrations returned to pre-treatment values, but cortisol fell to a level significantly lower. Regression correlations showed that Delta adrenaline and Delta scores, post-recovery, on the central executive task were significantly correlated. Delta noradrenaline correlated significantly with Delta fatigue. It was concluded that heat stress results in deterioration in the performance of central executive tasks and perceptions of mood state, and that this can be predicted by changes in body mass loss and plasma concentrations of the hormones cortisol and adrenaline. PMID- 16309772 TI - The properties of carbachol-induced beta oscillation in rat hippocampal slices. AB - The rhythmical and pharmacological properties of carbachol-induced beta oscillation were studied using rat hippocampal slices. With the application of 30 microM carbachol, beta-range oscillations with frequencies of 13-20 Hz were recorded from the CA3 region. An AMPA receptor antagonist, CNQX, diminished the oscillations. An NMDA receptor antagonist, APV, significantly suppressed the pre established beta oscillations. The pre-application of APV blocked the start of the carbachol-induced beta oscillations. When bicuculline (BIC), a GABAA receptor antagonist, was applied to the pre-established beta oscillations, the frequency decreased to the theta-range. When 5 microM BIC was applied with 30 microM carbachol, the beta oscillations did not start; instead, theta-like activities were induced. It has been reported that carbachol in hippocampal slices can induce theta-like activities, which are not modulated by BIC, while BIC's facilitating the start of the activities. The results of the present study suggest that the GABAA receptor-mediated inhibitory transmission modulates the beta oscillation and that the transmission is needed for the start process of the oscillations. Therefore, the start and generation mechanisms of carbachol-induced beta oscillation will be different from those of carbachol-induced theta-like activities. PMID- 16309773 TI - Chip calorimetry for the monitoring of whole cell biotransformation. AB - Efficient control of whole cell biotransformation requires quantitative real-time information about the thermodynamics and kinetics of growth and product formation. Heat production contains such information, but its technical application is restricted due to the high price of calorimetric devices, the difficulty of integrating them into existing bio-processes and the slow response times of established microcalorimeters. A new generation of chip or nanocalorimeters may overcome these weaknesses. We thus tested a highly sensitive chip calorimeter for its applicability in biotechnological monitoring. It was used to monitor aerobic growth of suspended and immobilized Escherichia coli DH5alpha DSM 6897 and anaerobic growth of suspended Halomonas halodenitrificans CCM 286(T). The chip data corresponded well with enthalpy balance calculations and measurements with a conventional calorimeter, indicating the applicability of the chip calorimeter for bio-process control. PMID- 16309774 TI - Ultrasound-mediated gene transfer into neuronal cells. AB - A new field of gene transfer is emerging as a simple, effective means to drive the expression foreign genes in cells: ultrasound-mediated gene transfer or sonoporation. We report here that sonoporation is an effective means of gene transfer for cultured neurons, a cell type that has been difficult to transfect. Neuronal cell types that are effectively sonoporated include chick retinal neurons, chick dorsal forebrain, chick optic tectum, PC12 cells, rat cerebellar neurons and mouse hippocampal neurons. Depending on the type of cell and conditions of sonoporation the transfection efficacy was as high as 20%. Sonoporation of plasmid DNA was effective for cells adherent to a substrate and for free-floating cells that were freshly dissociated. In the free-floating preparations, between 60 and 95% of the cells that were transfected were neuronal, as much as 90% higher than that observed for other methods of gene transfer including adenovirus and lipid-based transfection methods. We conclude that sonoporation is a simple, effective and inexpensive means by which to preferentially transfect DNA into neuronal cells. PMID- 16309775 TI - An oral delivery device based on self-folding hydrogels. AB - A self-folding miniature device has been developed to provide enhanced mucoadhesion, drug protection, and targeted unidirectional delivery. The main part of the device is a finger like bilayered structure composed of two bonded layers. One is a pH-sensitive hydrogel based on crosslinked poly(methyacrylic acid) (PMAA) that swells significantly when in contact with body fluids, while the other is a non-swelling layer based on poly(hydroxyethyl methacrylate) (PHEMA). A mucoadhesive drug layer is attached on the bilayer. Thus, the self folding device first attaches to the mucus and then curls into the mucus due to the different swelling of the bilayered structure, leading to enhanced mucoadhesion. The non-swelling PHEMA layer can also serve as a diffusion barrier, minimizing any drug leakage in the intestine. The resulting unidirectional release provides improved drug transport through the mucosal epithelium. The functionality of this device is successfully demonstrated in vitro using a porcine small intestine. PMID- 16309776 TI - An intracellular kinase signal-responsive gene carrier for disordered cell specific gene therapy. AB - We have previously reported artificial gene-regulation systems responding to cyclic AMP-dependent protein kinase (PKA) using cationic polymer. This cationic polymer (PAK) was a graft-type polymer with an oligopeptide that is a substrate for PKA and could regulate gene-expression in a cell-free system. In the present study, we carried out a detailed characterization of the PAK-DNA complex (AFM observation and DLS measurement) and tried to apply this polymer to living cells. In the unstimulated NIH 3T3 cells, transfection of the PAK-DNA complex showed no expression of the delivered gene. This means that PAK formed a stable complex with DNA in the normal cells to totally suppress gene expression. In contrast, significant expression was seen when the PAK-DNA complex was delivered to forskolin-treated cells. Thus, activated PKA disintegrates the complexes even in living cells, resulting in gene expression. Our results indicate that this type of intracellular signal-responsive polymer will be useful for the cell-specific release of genes. PMID- 16309777 TI - Is early and late post-meal exercise so different in type 1 diabetic lispro users? AB - To compare blood glucose (BG) responses during a 60 min moderate intensity exercise session performed in early or late postprandial periods. Nine generally well-controlled (HbA(1c): 7.3+/-0.1%) type 1 diabetic patients performed, at least one week apart, two exercise sessions, 60 (early exercise) and 180 min (late exercise) after a standardized breakfast. All subjects were using Humulin N (N) and Humalog (Lispro, LI) insulin. During exercise, the overall decrease in BG was 4.8+/-0.6 mmol/l and 3.6+/-0.8 mmol/l in early and late exercise, respectively (P=0.051). To prevent hypoglycemia, a dextrose infusion was initiated when BG reached 5 mmol/l. The quantity of dextrose infused was 6.2+/ 3.0 g and 10.5+/-3.2g in early and late exercise, respectively (NS). The time free of dextrose infusion during exercise was 41.2+/-7.8 min and 31.7+/-7.5 min in early and late exercise, respectively (NS). In N-LI users, overall drop in BG during exercise tends to be greater in the early postprandial period. However, early and late exercise present similar quantity of dextrose infused and time free of dextrose infusion. Consequently, the similar risk of exercise-induced hypoglycemia suggests similar precautions in either exercise times. PMID- 16309778 TI - Global synthetic-lethality analysis and yeast functional profiling. AB - The Saccharomyces genome-deletion project created >5900 'molecularly barcoded' yeast knockout mutants (YKO mutants). The YKO mutant collections have facilitated large-scale analyses of a multitude of mutant phenotypes. For example, both synthetic genetic array (SGA) and synthetic-lethality analysis by microarray (SLAM) methods have been used for synthetic-lethality screens. Global analysis of synthetic lethality promises to identify cellular pathways that 'buffer' each other biologically. The combination of global synthetic-lethality analysis, together with global protein-protein interaction analyses, mRNA expression profiling and functional profiling will, in principle, enable construction of a cellular 'wiring diagram' that will help frame a deeper understanding of human biology and disease. PMID- 16309779 TI - Mismatch repair converts AID-instigated nicks to double-strand breaks for antibody class-switch recombination. AB - Mismatch repair (MMR) proteins are important for antibody class-switch recombination (CSR), but their roles are unknown. We propose a model for the function of MMR in CSR in which MMR proteins convert single-strand nicks instigated by activation-induced cytidine deaminase (AID) into the double-strand breaks (DSBs) that are required for CSR. This model does not invoke any novel functions for MMR but simply posits that, owing to numerous single-strand nicks in the switch (S) regions of both DNA strands, when MMR proteins are recruited by U:G mismatches, they excise one strand of DNA and soon reach a nick on the opposite strand. This halts excision activity and creates a DSB. This model explains why B cells that lack either S mu and MSH2 or UNG and MSH2 cannot undergo CSR. PMID- 16309780 TI - On a chromosome far, far away: LCRs and gene expression. AB - Transcription activation of a gene involves the ordered recruitment of components of the basal transcription machinery in concert with alterations in chromatin structure, including nucleosome remodeling and post-translational modification of histones. Enhancers and locus control regions (LCRs) that are remote from the genes they activate, recruit the complexes that carry out these alterations and, sometimes, recruit RNA polymerase II. The question of how these distant activators interact with their target genes has been of long-standing interest. Recent data indicate that LCRs mediate contact with their coordinate genes through the formation of domains of histone modification and of intra- and inter chromosomal loops and that they might localize genes within nuclear regions that favor transcription. PMID- 16309781 TI - Wet age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of irreversible visual loss in industrialized nations for those age 65 and above. The majority of patients with severe visual loss suffer from the wet form of AMD wherein there is choroidal neovascularization (CNV) and associated manifestations such as retinal pigment epithelial detachment, subretinal hemorrhages, and fibrovascular disciform scarring. The main focus on understanding the pathogenesis of CNV has been on the hypothesis that the diffuse thickening of Bruch's membrane predisposes it to develop cracks and in-growth of new vessels from choriocapillaries with associated low-grade inflammatory response. Currently, three types of treatments (laser photocoagulation, photodynamic therapy, and anti Vascular Endothelial Growth Factor (VEGF) therapy) have been demonstrated to limit or delay loss of vision in patients. Only a minority of cases show stabilization of vision and a small proportion of cases show significant improvement in vision. This highlights the need for more and better pharmacologic or other interventions, with the goal of lowering recurrence rates and preventing the development of CNV in order to achieve better functional outcomes. PMID- 16309782 TI - Urinary catheterization in care homes for older people: self-reported questionnaire audit of catheter management by care home staff. AB - A self-administered questionnaire was used to determine care home staff's reported knowledge of the urinary catheter care standards published by the National Institute for Clinical Excellence (NICE) and the Association of Continence Care, and to see whether this differed in homes with higher catheterization rates. Seven hundred and fifty out of 1438 (52%) nursing and care staff from 37 randomly selected care homes with high, medium and low catheterization rates responded. There was no difference in reported practice in care homes in the three health districts sampled or those with differing catheterization rates. Eighty-three percent of the nursing staff and 40% of the other care staff received formal catheter care training. However, at least 10% of all staff reported not washing their hands before handling a catheter, and delaying emptying a urine bag until it was full, rather than three-quarters full. Only 45% of nursing staff and 40% of other care staff encouraged residents to empty their own catheter bags. Routine use of catheter maintenance solutions or bladder washouts was reported by 50% of all staff. Nursing staff (29%) and other care staff (54%) took urine specimens from the catheter bag tap. Compliance with standards has improved greatly since an audit in 1998. However, some non compliance remains. There is a need for ongoing local audit and formal training in urinary catheter care, particularly for non-qualified care staff. Education is needed to ensure local implementation of NICE guidance. PMID- 16309783 TI - Plan-do-study-act cycles as an instrument for improvement of compliance with infection control measures in care of patients after cardiothoracic surgery. AB - The aim of this study was to determine whether compliance with infection control measures for the care of patients during and after cardiothoracic surgery could be improved by using 'plan-do-study-act' (PDSA) improvement cycles in a 715-bed university hospital. The endpoints of these cycles were indices of correct procedure based on infection control standards. The intervention consisted of instruction and training of nursing and medical staff on the use of PDSA cycles, feedback of the baseline measurements, and the use of posters in the proximity of the operating room (OR). At follow-up, overall compliance only improved in the room used by the perfusionists and the OR. After the follow-up period, monitoring revealed a drop in compliance in the OR, but improved compliance during vascular catheter care of patients with prolonged stay in the intensive care unit (ICU), and during wound care of patients on the nursing ward. The last series of monitoring showed that compliance with general infection control measures in the OR had improved again, and that compliance had remained satisfactory on the ward and in the ICU, with the exception of patients recently transferred to the ICU from the OR. The results show that by using PDSA cycles, compliance with infection control measures can improve significantly. However, repeated monitoring is necessary to ensure continued compliance. PMID- 16309784 TI - Newly identified water-borne protein pheromones interact with attractin to stimulate mate attraction in Aplysia. AB - The water-borne protein attractin is a potent sex pheromone involved in forming and maintaining mating and egg-laying aggregations in the marine mollusk Aplysia. Binary blends of attractin and either enticin, temptin, or seductin, three other Aplysia protein pheromones, stimulate mate attraction. The four pheromones are thought to act in concert during egg-laying. The new data presented here show that: (1) the water-borne odor of non-laying Aplysia brasiliana further increases the attractiveness of attractin and of eggs in T-maze bioassays. This suggests that individual Aplysia release additional factors that enhance the effects of attractin, enticin, temptin, and seductin during egg-laying; (2) the N-terminal region of enticin aligns well with the conserved epidermal growth factor (EGF) like domain of mammalian reproductive proteins known as fertilins, which may mediate intercellular adhesion interactions between eggs and sperm; (3) temptin, according to fold recognition servers, may also have an EGF-like fold. Enticin and temptin also have conserved metal binding sequences that may play a role in their signaling behavior. These results suggest that aspects of mammalian egg sperm interactions (fertilins) may have evolved from pheromonal signaling mechanisms. We also review the structure, expression, localization, release, and behavioral actions of attractin, enticin, temptin, and seductin. PMID- 16309785 TI - Proctolin: A possible releasing factor in the corpus cardiacum/corpus allatum of the locust. AB - The corpus cardiacum (CC) and corpus allatum (CA) of the locust, Locusta migratoria, contain intense proctolin-like immunoreactivity (PLI) within processes and varicosities. In contrast, in the cockroach, Diploptera punctata, although a similar staining pattern occurs within the CC, PLI appears absent within the CA. The possible role of proctolin as a releasing factor for adipokinetic hormone (AKH) and juvenile hormone (JH) was investigated in the locust. Proctolin caused a dose-dependent increase in AKH I release (determined by RP-HPLC) from the locust CC over a range of doses with threshold above 10(-8)M and maximal release at about 10(-7)M proctolin. Isolated glandular lobes of the CC released greater amounts of AKH I following treatment with proctolin and in these studies AKH II was also released. Confirmation of AKH I release was obtained by injecting perfusate from incubated CCs into locusts and measuring hemolymph lipid concentration. Perfusate from CC incubated in proctolin contained material with similar biological activity to AKH. Proctolin was also found to significantly increase the synthesis and release of JH from locust CA, with the increase being greatest from CAs that had a relatively low basal rate of JH biosynthesis (<35 pmol h(-1) per CA). In contrast, proctolin did not alter the synthesis and release of JH from the cockroach CA. These results suggest that proctolin may act as a releasing factor for AKHs and JH in the locust but does not act as a releasing factor for JH in the cockroach. PMID- 16309786 TI - Melanocortin-5 receptor: a marker of human sebocyte differentiation. AB - Melanocortin receptors (MC1R-MC5R) and their ligands (melanocyte-stimulating hormone (MSH) and adrenocorticotrophin hormone (ACTH)) have been shown to influence physiological functions of cells and organs, including exocrine glands. Since relatively little is known about MC5R expression and function in the human sebaceous gland, we examined expression of MC5R by immunohistochemistry and RT PCR in human sebaceous cells in vivo and in vitro. In human skin, MC5R was detected only in differentiating, lipid-laden sebaceous cells but not in basal, undifferentiated sebaceous cells. Similarly, in cultured human sebocytes MC5R was only detectable at the onset of differentiation and in fully differentiated cells displaying prominent lipid granules. The lipid profile of the cultured and differentiated human sebocytes was shown to be human sebum-specific using (14)C acetate labeling and high performance thin layer chromatography. Our studies suggest that MC5R is a marker of human sebocyte differentiation. PMID- 16309787 TI - Proctolin-like immunoreactivity in the central and peripheral nervous systems of the locust, Locusta migratoria. AB - Proctolin-like immunoreactivity (PLI) was widely distributed in the locust, Locusta migratoria, within the central, peripheral and stomatogastric nervous systems, as well as the digestive system and retrocerebral complex. Proctolin like immunoreactivity was observed in cells and processes of the brain and all ganglia of the ventral nerve cord. Of interest, PLI was found in the lateral neurosecretory cells, which send axons within the paired nervi corporis cardiaci II (NCC II) to the corpus cardiacum (CC). The CC contained extensive processes displaying PLI, which continued on within the paired nervi corporis allata (NCA) to the paired corpora allata (CA) where the axons entered and branched therein. The frontal and hypocerebral ganglia of the stomatogastric nervous system contained PLI within processes, resulting in a brightly staining neuropile. Each region of the gut contained PLI in axons and processes of varying patterns and densities. The paired ingluvial ganglia contained PLI, including an extensively stained neuropile and immunoreactive axons projecting through the nerves to the foregut. The hindgut contained PLI within longitudinal tracts, with lateral projections originating from the 8th abdominal ganglion via the proctodeal nerve. The midgut contained PLI in a regular latticework pattern with many varicosities and blebs. No difference in PLI in cells and processes of the central nervous system (CNS) was found between males and females. PMID- 16309788 TI - Elevated maternal cortisol early in pregnancy predicts third trimester levels of placental corticotropin releasing hormone (CRH): priming the placental clock. AB - The purposes of this study were to determine the intervals when placental corticotrophic-releasing hormone (CRH) was most responsive to maternal cortisol. A sample of 203 women each were evaluated at 15, 19, 25 and 31 weeks gestation and followed to term. Placental CRH and maternal adrenocorticotropin hormone (ACTH), B-endorphin and cortisol were determined from plasma. CRH levels increased faster and were higher in women who delivered preterm compared with women who delivered at term (F3,603 = 5.73, p < .001). Simple effects indicated that CRH levels only at 31 weeks predicted preterm birth (F1,201 = 5.53, p = .02). Levels of cortisol were higher in women who delivered preterm at 15 weeks gestation (F1,201 = 4.45, p = .03) with a similar trend at 19 weeks gestation. Hierarchical regression suggested that the influence on birth outcome of maternal cortisol early in pregnancy was mediated by its influence on placental CRH at 31 weeks. Elevated cortisol at 15 weeks predicted the surge in placental CRH at 31 weeks (R = .49, d.f. = 1,199, Fchange = 61.78, p < .0001). Every unit of change in cortisol (microg/dl) at 15 weeks was associated with a 34 unit change of CRH (pg/ml) at 31 weeks. These findings suggested that early detection of stress signals by the placenta stimulated the subsequent release of CRH and resulted in increased risk for preterm delivery. PMID- 16309789 TI - Novel excitatory neuropeptides isolated from a prosobranch gastropod, Thais clavigera: The molluscan counterpart of the annelidan GGNG peptides. AB - The GGNG peptides are excitatory neuropeptides identified from earthworms, leeches and polychaeta. Two structurally related peptides were purified and characterized from a mollusk, Thais clavigera (prosobranch gastropod). The peptides designated as Thais excitatory peptide-1 (TEP-1) (KCSGKWAIHACWGGN-NH2) and TEP-2 (KCYGKWAMHACWGGN-NH2) are pentadecapeptides having one disulfide bond and C-terminal GGN-NH2 structures, which are shared by most GGNG peptides. TEP augmented the motilities of Thais esophagus and penial complex. TEP-like immunoreactivity is distributed in both the neurons of the central nervous system and nerve endings in the penial complex. Thus, the involvement of TEP in the contraction of the digestive and reproductive systems is suggested. Substitution of amino acids in TEP revealed that two tryptophan residues in TEP are important for maintaining bioactivity. PMID- 16309790 TI - A comparison of the pupariation acceleration activity of pyrokinin-like peptides native to the flesh fly: Which peptide represents the primary pupariation factor? AB - Five native pyrokinin-like peptides (Neb-PK-1, Neb-PK-2, Neb-PVK-1, [L9]Neb-PVK 2, [I9]Neb-PVK-2) identified in the neuropeptidome of the flesh fly Neobellieria bullata were compared for their quantitative and/or qualitative effects on puparium formation (pupariation). In a standard pupariation bioassay, both Neb PVK-1 and [I9]Neb-PVK-2 proved inactive, whereas [L9]Neb-PVK-2 demonstrated only weak activity. In contrast, both Neb-PK-1 and Neb-PK-2 demonstrated potent threshold doses, with Neb-PK-2 about 10-fold more active than Neb-PK-1. Analysis of neuromuscular activity during pupariation using a tensiometric technique demonstrates that the two native Neb-PKs accelerate the onset of immobilization and cuticular shrinkage more than motor programs associated with retraction of the anterior segments and longitudinal body contraction. It was further determined that the sensitivity of various components of the pupariation process to these peptides decreases in the following order: immobilization>cuticular shrinkage>motor program for anterior retraction>motor program for longitudinal contraction congruent to tanning of cuticle of the newly formed puparium. A paradoxical situation was observed whereby the motor programs of pupariation are temporally dissociated from actual morphogenesis of the puparium. The tensiometric data suggest that the most likely candidate for a primary pupariation factor is Neb-PK-2, rather than Neb-PK-1. PMID- 16309791 TI - Analysis of peptides in the brain and corpora cardiaca-corpora allata of the honey bee, Apis mellifera using MALDI-TOF mass spectrometry. AB - The neuropeptide profiles and diversity of the brain and retrocerebral organs (corpora cardiaca-corpora allata; CC-CA) of adult workers of the honey bee Apis mellifera carnica (dark European strain) were investigated using a combination of HPLC and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) with post-source decay (PSD) and collision-induced dissociation (CID) fragmentation. Using evidence from genomic sources, including BLAST searches of the honey bee genome, comparisons with other species and de novo sequencing by PSD and CID fragmentation, a total of 13 mass ions could be assigned to peptides predicted from the A. mellifera genomic database. Peptides positively identified were A. mellifera tachykinin-related peptides 3 and 4 (APMGFQGMRa; APMGFYGTRa) and leucomyosuppressin (pEDVDHVFLRFa). Peptides tentatively identified were A. mellifera tachykinin-related peptides 2 and 5 (ALMGFQGVRa; ARMGFHGMRa), A. mellifera allatostatins 2, 3 and 4 (GRDYSFGLa; RQYSFGLa; GRQPYSFGLa), A1-SIFamide (AYRKPPFNGSIFa), Q1-leucomyosuppressin (QDVDHVFLRFa) and A. mellifera pyrokinins PK 1, PK 2 and Q1-PK 2 (TSQDITSGMWFGPRLa; pEITQFTPRLa; QITQFTPRLa). Allatostatins, tachykinin-related peptides and A1-SIFamide were not detected in CC-CA extract, which appears to contain predominantly leucomyosuppressin, Q1-leucomyosuppressin, PK 1, PK 2, Q1 PK 2 and some unidentified masses. No ion signal was detected that would correspond to the hypertrehalosaemic peptide (=Manse-AKH), which has been isolated from the Italian race of the honey bee (A. mellifera ligustica), but not from A. mellifera carnica. PMID- 16309792 TI - Interactions between gut peptides and the central melanocortin system in the regulation of energy homeostasis. AB - Genetic and pharmacological studies have shown that the central melanocortin system plays a critical role in the regulation of energy homeostasis. Animals and humans with defects in the central melanocortin system display a characteristic melanocortin obesity phenotype typified by increased adiposity, hyperphagia, metabolic defects and increased linear growth. In addition to interacting with long-term regulators of energy homeostasis such as leptin, more recent data suggest that the central melanocortin system also responds to gut-released peptides involved in mediating satiety. In this review, we discuss the interactions between these systems, with particular emphasis on cholecystokinin (CCK), ghrelin and PYY(3-36). PMID- 16309793 TI - Characterization and localization of cocaine- and amphetamine-regulated transcript (CART) binding sites. AB - Cocaine- and amphetamine-regulated transcript (CART) is widely expressed in the brain and various endocrine tissues. CART is implicated in many physiological functions including food intake, drug reward, stress and nociception. No CART receptor has been identified yet. We fused CART(55-102) to the green fluorescent protein (GFP) and found that the ligand suppresses significantly food intake after intracerebroventricular (i.c.v.) injection in mice. Using this ligand, we show specific CART binding sites on HepG2 cells and hypothalamic dissociated cells. In brain sections, CART displaceable binding sites were observed on cell bodies mainly localized in hypothalamic periventricular areas. PMID- 16309794 TI - Degradation profile of [His7]-corazonin in the hemolymph of the desert locust Schistocerca gregaria. AB - Degradation of the neuropeptide [His7]-corazonin, a key hormone in phase transition in locusts was studied using [3H][His7]-corazonin, RP-HPLC and mass spectrometry. After 4h incubation, 50 and 75% of [His7]-corazonin could still be found in hemolymph of gregarious and solitarious Schistocerca gregaria, respectively. Under in vivo conditions the half-life was 30 min. These results are in contrast to many other neuropeptides that usually have half lives of a few minutes. The peptide is cleaved first by an endopeptidase, either just before or after the Tyr residue at position 5. Next, the C-terminal degradation fragments are further degraded by a dipeptidyl-peptidase, whereas the N-terminal fragments are further broken down one amino acid at a time. In addition, [Dopa5][His7] corazonin was detected. Upon synthesis, this unexpected molecular modification turned out to be biologically active in bringing about cuticular melanization. PMID- 16309795 TI - Volume reduction of the entorhinal cortex in subjective memory impairment. AB - To examine the biological basis of subjective memory impairment (SMI), defined as the feeling of memory worsening with normal memory performance, we measured the volume of the entorhinal cortex (EC) and the hippocampus in SMI subjects, patients with mild cognitive impairment (MCI), patients with Alzheimer's disease (AD) and healthy controls (CO). Compared with controls, the EC was smaller in the SMI group (left: p=0.060; right: p=0.045) and in the other two groups in the following order: CO>SMI>MCI>AD. The same sequence was observed with regard to hippocampal volumes, but the volume reduction of the left hippocampus in the SMI group only reached a trend towards significance (p=0.072) and the right was not significantly smaller compared with controls (p=0.37). Compared with controls the average (left/right) volume reduction of the EC was 18% (SMI), 26% (MCI) and 44% (AD). The mean volume reduction of the hippocampus was 6% (SMI), 16% (MCI) and 19% (AD). Our results mirror the temporal sequence of neurodegeneration in AD and support the concept of SMI as the first clinical manifestation of dementia. PMID- 16309796 TI - Beneficial effects of regular exercise on sleep in old F344 rats. AB - With aging there is a significant decline in the normal architecture of sleep and a reduction in the diurnal amplitude of core body temperature. Regular moderate exercise has been shown to have a positive impact in the elderly and here we investigate whether sleep-wake patterning can also be improved. Young (3 months) and old (22 months) F344 rats were exercised once a day for 50min at night onset over an 8-week period. Thereafter, polysomnographic recordings were obtained immediately after exercise. To determine the lasting consequences of exercise, sleep was also recorded 2 days and 2 weeks after exercise had ended. Old rats that were exercised had a significant weight loss, were awake more during the last third of their active period, had less sleep fragmentation and the amplitude of the diurnal rhythm of core body temperature was significantly increased. Old exercised rats also had an overall increase in the amplitude of EEG power (0.5 16Hz) during wake and theta EEG power during REM sleep. In young rats regular exercise increased EEG delta power (0.5-4Hz) during NREM sleep. Our data indicate regular exercise in old rats improves sleep architecture, EEG power and diurnal rhythm of temperature. PMID- 16309797 TI - Responsiveness of the aging circadian clock to light. AB - The present study assessed whether advances in sleep times and circadian phase in older adults might be due to decreased responsiveness of the aging circadian clock to light. Sixteen young (29.3+/-5.6 years) and 14 older adults (67.1+/-7.4 years) were exposed to 4h of control dim (10lux) or bright light (3500lux) during the night. Phase shifts of the melatonin rhythm were assessed from the nights before and after the light exposure. Bright light delayed the melatonin midpoint in both young and older adults (p<0.001). Phase delays for the older subjects were not significantly different from those of the young subjects for either the bright or dim light conditions. The magnitude of phase delays was correlated with both sleep offset and phase angle in the older, but not the younger subjects. The present results indicate that at light intensities commonly used in research as well as clinical practice older adults are able to phase delay to the same extent as younger subjects. PMID- 16309798 TI - [Pentoxifylline: a cheap substitute for anti-TNFalpha agents?]. AB - PURPOSE: Pentoxifylline (PTX) is a phosphodiesterase inhibitor drug used to improve peripheral vascular disease. In vitro studies demonstrated that PTX has anti-TNFalpha properties. We did a selective review of clinical trials which used PTX in patients with inflammatory rheumatic and non-rheumatic diseases. CURRENT KNOWLEDGE AND KEY POINTS: Most of the identified clinical trials were uncontrolled and involved a low number of patients. Use of PTX in systemic lupus erythematous, Behcet's disease and sarcoidosis yielded significant preliminary results. Moreover, PTX markedly reduced proteinuria in several glomerulonephritis (lupus nephritis, membranous nephropathy, diabetic nephropathy). FUTURE PROSPECTS AND PROJECTS: Further randomized and controlled clinical trials are required to examine whether PTX can improve outcome in patients with inflammatory diseases. Meanwhile, PTX should not be used for the treatment of these diseases. PMID- 16309799 TI - [Conjugal progressive systemic sclerosis]. AB - INTRODUCTION: Familial occurrence of progressive systemic sclerosis is unusual. The occurrence of conjugal scleroderma is exceptional. EXEGESIS: We report here a case of systemic sclerosis in a wife and husband who both developed the onset of illness within a 10-year period. Solvent exposure was noted. CONCLUSION: The etiology of systemic sclerosis remains unknown. Environmental factors may play role in its pathogenesis. PMID- 16309800 TI - Knowing nursing and finding the professional voice: a study of enrolled nurses converting to first level registration. AB - This study explored the experiences of a group of second level registered nurses converting to first level registration in a two-year, part-time open learning programme. It aimed to examine the relationship between the mode of course delivery and the personal and professional development experienced by the learners. Data were collected by semi-structured interviews, conducted at five points over the two-year period and analyzed by constant comparative analysis. A theoretical model was developed to demonstrate the stages of knowing nursing, experienced by the students as they made the transition from passive to autonomous professional practice. In their 'Women's Ways of Knowing' [Belenky M.F., Clinchy B.A., Goldberger N.R., Tarule J.M., 1986. Women's Ways of Knowing. Basic Books, New York] identified a progression through different stages of thinking about the nature of knowledge and evidence. This progression was evident in the nurses in this study. It was applied and developed to reflect the progression through ways of knowing nursing. This process consisted of three positions of development, each underpinned by their associated ways of knowing [Belenky M.F., Clinchy B.A., Goldberger N.R., Tarule J.M., 1986. Women's Ways of Knowing. Basic Books, New York] and characterized by a specific approach to professional nursing practice. The changing ways of knowing nursing, described in this paper, are likely to be observed in registered nurses and health care assistants who are now entering Higher Education (HE) to undertake higher awards for the first time. A clearer understanding of this process of personal and professional change, and factors that impact upon it, will be a useful guide for those supporting learners both in the clinical and academic arenas. PMID- 16309801 TI - Serological and phylogenetic evidence of monotypic immune responses to different mumps virus strains. AB - The recent resurgence of mumps epidemics in many countries with ongoing vaccination programs along with evidence of antigenic diversity among mumps virus strains have recently challenged the assumption that mumps virus is serologically monotypic. To address this controversy, we sought to identify two mumps virus strains that would best represent different serotypes, should multiple serotypes exist, and assess the ability of human sera to neutralize both strains. The virus strains, Enders and Lo1, were selected based upon a phylogenetic analysis of the major target of neutralizing antibody, the viral hemagglutinin-neuraminidase (HN) protein, along with data reported by others indicating that (1) these viruses are antigenically distinct and (2) genotypically similar strains have been implicated in cases of reinfection. Our results show that of sera capable of neutralizing one of the virus strains, 90% could neutralize the other, although significant differences in neutralization titers were noted. Though the latter confirms the existence of inter-strain antigenic variability, the fact that few sera were unable to neutralize both virus strains argues against the presence of multiple serotypes. Of those sera incapable of co-neutralization, all but one had low neutralization titers (1:8), suggesting that individuals possessing low levels of neutralizing antibody may be at risk for breakthrough infections, thereby providing an explanation for cases of infection in previously infected or vaccinated individuals. PMID- 16309802 TI - INTERESTING CASE: acoustic neuroma--an unusual presentation. PMID- 16309803 TI - INTERESTING CASE: an interesting fracture of the zygomatic arch. PMID- 16309804 TI - Heavy metals affect the coelomocyte-bacteria balance in earthworms: environmental interactions between abiotic and biotic stressors. AB - Three-day dermal exposure of Dendrobaena veneta to metal ions differentially disrupts the immunocompetence/pathogen balance. Zn does not accumulate in the earthworm body, Cu accumulation is temperature-independent while Cd accumulation is stronger at 22 degrees C than at 10 degrees C. During in vitro incubation with metal ions at 22 degrees C, growth of coelom-derived bacteria is enhanced by Zn, but significantly or almost completely inhibited by Cu or Cd. In contrast, under in vivo conditions at 22 degrees C, bacterial load is decreased only after Cd exposure, but increased after Zn and Cu exposures. At 10 degrees C bacteria growth is almost completely inhibited in all groups except Cu-treated animals. Coelomocyte number is unaffected in animals exposed to Zn, but significantly decreased after exposure to Cd (at 22 degrees C) and Cu (at 22 degrees C and 10 degrees C) with concomitant changes of amoebocyte-to-eleocyte ratio in favour of amoebocytes. Metal exposure up-regulates expression of metallothioneins in coelomocytes, mainly amoebocytes. PMID- 16309805 TI - The performance of health workers in Ethiopia: results from qualitative research. AB - Insufficient attention has been paid to understanding what determines the performance of health workers. This paper reports on findings from focus group discussions with both health workers and users of health services in Ethiopia, a country with some of the poorest health outcomes in the world. We describe performance problems identified by both health users and health workers participating in the focus group discussions, including absenteeism and shirking, pilfering drugs and materials, informal health care provision and illicit charging, and corruption. In the second part of the paper we present four structural reasons why these problems arise: (i) the ongoing transition from health sector dominated by the public sector, towards a more mixed model; (ii) the failure of government policies to keep pace with the transition towards a mixed model of service delivery; (iii) weak accountability mechanisms and the erosion of professional norms in the health sector; and (iv) the impact of HIV/AIDS. The discussions underline the need to base policies on a micro-analysis of how health workers make constrained choices, both in their career and in their day to day professional activities. PMID- 16309806 TI - Psychosocial problems of bereaved families of HIV-infected hemophiliacs in Japan. AB - This study sought to investigate the psychosocial difficulties faced by bereaved families of HIV-infected hemophiliacs in Japan, with the goal of clarifying appropriate ways to offer them support. Interviews were conducted with 46 family members from 36 families, and a written survey was mailed to 392 families. Valid responses from 225 families (307 family members) were analyzed (response rate 57.4%). We found that even at a mean 7-9 years after their family member's death, the bereaved continued to express deep feelings of sorrow and grief, and further, expressed strong feelings of resentment, anger, guilt and regret, anxiety over discrimination, and loneliness. Notably, 95% of the mothers endorsed feeling "sorry for giving birth to a hemophiliac child". Additionally, even after their family member's death, approximately 70% of the respondents continued to restrict their daily activities due to stigmatization or anxiety over discrimination. Finally, 59% of the respondents were found to have posttraumatic stress disorder like symptoms according to the Impact of Event Scale-Revised, and 46% were found, according to General Health Questionnaire-12 (a measure of mental disorder) criteria, to suffer mental health problems. These results indicate that bereaved families of HIV-infected hemophiliacs in Japan have special characteristics and psychosocial needs, and still suffer significant psychosocial difficulties years after the bereavement. PMID- 16309807 TI - Subjective response to clozapine and risperidone treatment in outpatients with schizophrenia. AB - The purpose of the present study was to compare the subjective response and attitude towards antipsychotic treatment between schizophrenic patients receiving clozapine and those receiving risperidone. Ninety-four outpatients who had been on a stable drug dosage were evaluated (clozapine group: n=57, mean dose=254.1 mg/day; risperidone group: n=37, mean dose=3.0 mg/day). Subjective response to antipsychotic treatment was assessed using the Drug Attitude Inventory (DAI). The two treatment groups had a positive total mean score, indicating that both groups had a positive subjective view of drug treatment. The proportion of subjects who had a positive total score was not different between the two groups. In subscale scores, multivariate analysis revealed that clozapine group tended to have a higher score on the subjective positive response subscale (P=0.06). The scores of subjective negative response or attitude to medication subscales were not different between groups. In conclusion, there was no marked difference between stabilized outpatients taking clozapine and risperidone in terms of subjective response and attitude towards antipsychotic treatment. Considering that subjects treated with clozapine were treatment resistant patients, equal DAI score might indicate a more favorable subjective experience of clozapine. Further prospective studies on subjective response to various atypical agents are required to obtain valuable insight into how best to use these drugs from the patient's perspective. PMID- 16309808 TI - Does domperidone potentiate mirtazapine-associated restless legs syndrome? AB - There is now evidence to suggest a central role for the dopaminergic system in restless legs syndrome (RLS). For example, the symptoms of RLS can be dramatically improved by levodopa and dopamine agonists, whereas central dopamine D2 receptor antagonists can induce or aggravate RLS symptoms. To our knowledge, there is no previous report regarding whether domperidone, a peripheral dopamine D2 receptor antagonist, can also induce or aggravate symptoms of RLS. Mirtazapine, the first noradrenergic and specific serotonergic antidepressant (NaSSA), has been associated with RLS in several recent publications. The authors report here a depressed patient comorbid with postprandial dyspepsia who developed RLS after mirtazapine had been added to his domperidone therapy. Our patient started to have symptoms of RLS only after he had been treated with mirtazapine, and his RLS symptoms resolved completely upon discontinuation of his mirtazapine. Such a temporal relationship between the use of mirtazapine and the symptoms of RLS in our patient did not support a potentiating effect of domperione on mirtazapine-associated RLS. However, physicians should be aware of the possibility that mirtazapine can be associated with RLS in some individuals, especially those receiving concomitant dopamine D2 receptor antagonists. PMID- 16309809 TI - A preliminary randomized double blind clinical trial on the efficacy of aqueous extract of Echium amoenum in the treatment of mild to moderate major depression. AB - BACKGROUND: Based on a traditional belief, Echium amoenum (Boraginaceae) dried flowers are used in Iran as an anxiolytic remedy and also as a mood enhancer. In this study, efficacy of an aqueous extract of E. amoenum in patients with mild to moderate major depressive disorder (a score > or =18 on the Hamilton depression rating scale) was evaluated. METHODS: 35 patients were randomly assigned to receive daily either placebo or 375 mg of E. amoenum aqueous extract in a 6-week double blind, parallel-group trial. Patients were assessed in weeks 0, 1, 2, 4 and 6 by the Hamilton Rating Scale for Depression (HAM-D17), the Hamilton Rating Scale for Anxiety (HAM-A14), and a score sheet on adverse effects. RESULTS: In week 4, the extract showed a significant superiority over placebo in reducing depressive symptoms. The effect on anxiety was not significant. Headache, somnolence, vomiting, dry mouth, constipation and blurred vision are the most commonly reported side effects. However, with regards to these side effects, no significant difference between placebo and drug treated groups was observed. LIMITATIONS: This study is performed with a small sample size, only with one dose and for 6 weeks. CONCLUSION: E. amoenum aqueous extract may have some antidepressant activity. PMID- 16309810 TI - Buprenorphine treatment outcome in dually diagnosed heroin dependent patients: A retrospective study. AB - The present study compared retrospectively in a clinical non-experimental setting the efficacy of buprenorphine (BUP) in different subgroups of dually diagnosed and non-dually diagnosed opioid-dependent patients: all the subjects included in the study showed severe long-lasting heroin addiction and 68.4% were affected by psychiatric comorbidity. Participants (206) (mean age 32.2+/-8.9, 177 males-29 females) were applicants to a long-term buprenorphine treatment program (mean doses 7.9+/-0.42 mg). Aim of the study was to evaluate dual diagnosis variables possibly influencing retention rate and abstinence from illicit drugs. The patients were divided into 5 subgroups on the basis of dual diagnosis: group 1: major depression (MD) 29.61%; group 2: generalized anxiety (GAD) (11.2%); group 3: personality disorders (PD), antisocial-borderline (21.84%); group 4: schizophrenia (SC)(6.3%); group 5: substance use disorder without overt psychiatric comorbidity (SUD) (31.1%). Group 1 patients affected by MD showed the highest retention rate at 12 months (72.1%) in comparison with the other groups of patients: group 2 GAD (39.1%), group 3 PD (17.8%), group 4 SC (7.7%) and group 5 SUD, without comorbidity (45.3%) (p=0.006, p<0.001, p<0.001, p=0.002). Similarly, at 12 months, the patients affected by MD showed less risk of illicit opioid use (16.4%) than those affected by GAD (34.8%), PD (42.2%), SC (53.8%) and SUD without comorbidity (34.4%) (p=0.06, p=0.003, p=0.008, p=0.017). When evaluated on the whole sample, retention rate was not influenced by dose. In contrast, the higher BUP doses were associated with less risk of illicit opioid use, than lower doses (p<0.001). Multivariate analysis and factor analysis showed a greater association of outcome measures (retention rate and negative urines rate) with comorbid diagnosis (depression) (respectively 0.64) than with buprenorphine doses (respectively 0.54). Our data need to be interpreted with caution because of the retrospective methodology applied to a clinical non experimental setting. BUP seems to be more effective in opioid-dependent patients affected by depression, probably due to the kappa opioid-receptors antagonist action, counteracting dysphoria, negativism and anxiety. High doses of BUP appear to predict a better outcome, in terms of negative urines, but not in terms of retention. PMID- 16309811 TI - Cadmium, lead, and thallium in mainstream tobacco smoke particulate. AB - The deliveries of cadmium, thallium, and lead in mainstream smoke particulate from cigarettes with different smoke delivery designs were determined by inductively coupled plasma-mass spectrometry in order to investigate their impact on the delivery of these known toxic compounds. Analyses showed that the levels of all three metals in smoke particulate were associated with their tar delivery category. After normalizing the metal concentrations to tar, there were no longer any statistically significant delivery differences between full-flavor, light or ultra-light cigarettes. When the concentrations were normalized to nicotine, the mean levels from the three delivery groups were much smaller than before normalization. But unlike the case using tar to normalize, in some of the cases, there were still some statistically significant differences in the nicotine normalized results. These findings suggest that if smokers compensate for differences in nicotine intake, they receive exposures to toxic heavy metals from ultra-light, light and full-flavor cigarettes that are more similar than results would suggest from using the Federal Trade Commission method alone. PMID- 16309812 TI - Induction of lupus-related specific autoantibodies by non-specific inflammation caused by an intraperitoneal injection of n-hexadecane in BALB/c mice. AB - A single intraperitoneal (i.p.) injection of pristane, incomplete Freund's adjuvant (IFA), or the adjuvant oil squalene, but not high molecular weight medicinal mineral oils, induces lupus-related autoantibodies to nRNP/Sm and -Su in non-autoimmune strains of mice. This ability appears to be associated with the low molecular weight and adjuvanticity of hydrocarbon. n-Hexadecane (C(16)H(34)), which is present in petroleum, has adjuvant activity and induces arthritis in rodents like other lupus-inducing oils. In addition to dietary exposure to n hexadecane in mineral oils, exposure also occurs via inhalation of oil mist, jet fuel, or diesel exhaust or by absorption through the skin. Since n-hexadecane is a low molecular weight adjuvant hydrocarbon oil similar to other lupus-inducing hydrocarbons, the present study examined whether it can also induce lupus-related autoantibodies in mice. Female BALB/cJ mice received a single i.p. injection of 0.5 ml of n-hexadecane, pristane, or saline (control). Pathology and serology (immunoglobulin levels, autoantibodies by immunofluorescence, immunoprecipitation, and ELISA) were examined 3 months later. Unexpectedly, all n hexadecane-treated mice, but none in the other groups, developed inflammatory ascites within 2.5 months. n-Hexadecane induced hypergammaglobulinemia (IgG1, IgG2a), antinuclear (titer>1:160, 67%) and -cytoplasmic antibodies (58%) and autoantibodies to nRNP/Sm (25%), Su (33%), ssDNA (83%), and chromatin (100%). Therefore, non-specific inflammation caused by n-hexadecane resulted in the production of a limited set of specific autoantibodies. These previously unrecognized immunological effects of n-hexadecane may have implications in monitoring human exposure to hydrocarbons and in the pathogenesis of autoimmune diseases. PMID- 16309813 TI - Comparison of chlordecone effects on autoimmunity in (NZBxNZW) F(1) and BALB/c mice. AB - It has been shown previously that chronic treatment with relatively low doses of chlordecone accelerates the development of systemic lupus erythematosus (SLE) in ovariectomized female (NZBxNZW) F(1) mice. In this study, the effect of chronic chlordecone treatment on SLE was evaluated in ovary-intact female (NZBxNZW) F(1) mice, as well as in female mice from a strain that is not lupus-prone, BALB/c. Chlordecone was administered chronically via implanted sustained-release pellets, and mice were monitored over time for the appearance of elevated autoantibodies (anti-dsDNA and anti-chromatin) and for the development of renal impairment, both indicators of SLE. Chlordecone treatment in (NZBxNZW) F(1) mice shortened significantly the time to onset of elevated autoantibody titers and renal disease in a dose-related manner. The doses required to produce this effect were similar to those observed previously to accelerate SLE development in ovariectomized females. Treatment of female BALB/c mice with chlordecone for up to one year did not produce elevated autoantibody titers or renal disease, suggesting an inability of chlordecone to cause a break in tolerance in this strain. These observations confirm the ability of chronic chlordecone to influence SLE, but demonstrate the importance of genetic background for this effect. PMID- 16309815 TI - Factors associated with self-reported halitosis in Kuwaiti patients. AB - OBJECTIVES: Oral malodor is a common complaint of dental patients, yet limited data is available on the actual prevalence of this condition. The aim of this study was to assess the prevalence and factors associated with self-reported halitosis in Kuwaiti patients. METHODS: This was a cross-sectional study of Kuwaiti adults using a 19-point self-administered structured questionnaire on self-perception of halitosis. Significant associations between self-reported oral malodor and sociodemographic, medical history, and oral hygiene variables were examined with multiple logistic regression analysis. RESULTS: A total of 1551 subjects participated (response rate=86.2%). The prevalence of self-reported halitosis was 23.3%. Use of the toothbrush less than once daily was the factor most strongly associated with self-perceived halitosis (OR=2.68; 95% CI=1.83 3.92; p<0.001). Other factors significantly associated with self-perceived halitosis included current or past smoking (OR=2.51), female gender (OR=1.54), being 30 years of age or older (OR=1.35), having high school education or less (OR=1.41), history of chronic sinusitis (OR=1.58) or gastrointestinal disorders (OR=1.73), never using miswak (OR=1.56), and never using dental floss (OR=1.33). CONCLUSION: Inadequate oral hygiene practices were the factors most strongly associated with self-reported oral malodor in this sample of Kuwaiti patients. Other factors with significant associations included history of gastrointestinal tract disorders, chronic sinusitis, older age, female gender, and lower education levels. PMID- 16309816 TI - Two domains are critical for the nuclear localization of soluble adenylyl cyclase. AB - Soluble adenylyl cyclase (sAC) is a newly identified source of cyclic adenosine 3',5'-monophosphate (cAMP). Unlike the well-known transmembrane adenylyl cyclases (tmACs), sAC locates to the nucleus, mitochondria and microtubules. For most cAMP signaling microdomains, there is always an AC nearby, for example tmAC. But it was until the discovery of sAC that there was not known cAMP resource in the nucleus. sAC associates with nuclear cAMP-signaling microdomains, which were once considered to depend on the diffusion of cAMP produced by tmAC. In this report, we focus on the truncated soluble adenylyl cyclase (tsAC), the most common existence form of sAC in tissues. Two domains (145-200 aa and 257-318 aa) related with sAC nuclear localization were present here. The findings provide evidence that these two domains are critical for the nuclear localization of sAC and they collocated with the catalytic domains. PMID- 16309817 TI - Adaptation to carbon starvation: RNase III ensures normal expression levels of bolA1p mRNA and sigma(S). AB - bolA is a sigma(S)-dependent Escherichia coli morphogene involved in the general cellular adaptation to stress and cell division. In this report it is shown that endoribonuclease RNase III acts as a post-transcriptional modulator of bolA expression under carbon starvation conditions. Unexpectedly RNase III positively regulates bolA1p mRNA levels and stabilities. This effect is also observed when sulA, bfr, uspA and uspB transcripts were analyzed. RNase III is furthermore shown to be necessary for the normal expression of sigma(S), ensuring normal levels of rpoS mRNA and sigma(S) protein under glucose starvation. Since sigma(S) controls a complex regulon of stress-response genes, RNase III is proposed as possible modulator of bacterial cell response to stress. PMID- 16309818 TI - COX-2 overexpression in peritoneal lesions is correlated with nonmenstrual chronic pelvic pain. AB - OBJECTIVE: To investigate cyclooxygenase (COX-2) expression within different endometriotic lesions and to assess whether these expression patterns correlate with clinical characteristics. DESIGN: Retrospective cross-sectional study. SETTING: University Hospital. PATIENTS: Seventy patients with histologically confirmed exclusively peritoneal (n=20), ovarian (n=19) or deep-infiltrating (n=31) endometriosis and a detailed medical history. INTERVENTION: Immunohistochemical analysis for COX-2 was performed on 108 endometriotic lesions. MEASUREMENTS AND MAIN RESULTS: COX-2 intensity, percentage of stained glandular endometriotic cells, and correlation of COX-2 expression with clinicopathological parameters. Semiquantitative COX-2 expression did not differ between distinct morphological types of endometriosis and showed no association with the menstrual cycle. Patients with peritoneal-only endometriosis suffering from moderate or severe chronic pelvic pain showed significantly more frequent COX-2 overexpression than asymptomatic patients or patients with minimal symptoms. In patients with exclusively ovarian or deep-infiltrating endometriosis no association between COX-2 expression and clinical parameters, such as chronic pelvic pain, dysmenorrhoea, dyspareunia, sterility, lower urinary tract symptoms or gastrointestinal symptoms was observed. CONCLUSION: Peritoneal endometriotic lesions with increased COX-2 expression have a special relevance for the development of chronic, nonmenstruation-associated, pelvic pain in endometriotic patients. These patients may benefit from therapy with COX-2 inhibitors. PMID- 16309819 TI - Placental site does not change background uterine electromyographic activity in the middle trimester of pregnancy. AB - OBJECTIVE: This study was performed in order to assess the potential influence of placental implantation site on transabdominal electromyographic (EMG) assessment of the uterine electrical activity in the middle trimester of pregnancy. STUDY DESIGN: In this prospective study 251 unselected, nulliparous asymptomatic women with a singleton pregnancy underwent transabdominal uterine EMG. Uterine electrical activity was recorded using bipolar electrodes placed on the abdominal surface for 20min. Regarding the placental implantation site and presence of action potentials (AP) pregnant women were divided into two groups: the anterior placenta group (APG) and the posterior placenta group (PPG). Outcome measures were differences in the median frequency (MF) and median amplitude (MA) of AP between the two groups. RESULTS: Action potentials were detected in 56 women: 33/56 in the APG versus 23/56 in the PPG. The parameters analyzed (MF, p=0.527, Fisher's exact test, and MA, p=0.255, Fisher's exact test) did not produce any statistical significant differences between the two groups. CONCLUSION: Background uterine EMG activity measured from the abdominal surface in the middle trimester of pregnancy does not depend on the placental implantation site. PMID- 16309820 TI - Human papillomavirus infection and cervical intraepithelial neoplasia in a cohort of low-risk women. AB - OBJECTIVE: This study was a prospective examination of the incidence of human papillomavirus (HPV) infection in a low-risk female population and an assessment of the risk of development of LSIL with HPV infection. STUDY DESIGN: In a longitudinal study, women aged 19-60 years--non-smokers, and married or living with a constant partner, who presented for cervical cancer screening at an outpatient clinic--were invited to participate in a prospective study of cervical HPV infection, and were examined every 3 months. RESULTS: Of the 464 women at risk, 20 presented with HPV infections during the follow-up. Low-grade squamous intraepithelial lesions (LSIL) event developed in 18 women. Among these women, 13 were HPV-positive (10 high-risk and 3 low-risk types). The average duration of new LSIL was 20.1 months (95% CI: 13.9-26.3) and 55.3 months (95% CI: 45.7-64.9) in the HPV-positive and negative groups, respectively, the difference was statistically significant (p<0.001). With the use of Cox proportional hazard regression, we estimated the relative risk as 90.0 for a first instance of LSIL among women testing positive for HPV as compared with women testing negative for HPV. CONCLUSION: This study has provided evidence that HPV infection is associated with an increased risk of LSIL. PMID- 16309821 TI - The use of transabdominal cervicoisthmic cerclage in successive pregnancies. PMID- 16309822 TI - Patients' obstetric history in mid-trimester termination of pregnancy with gemeprost: does it really matter? AB - OBJECTIVE: The objective was to investigate the importance of previous obstetric history for termination of pregnancy in the second-trimester with gemeprost alone. STUDY DESIGN: A consecutive series of 423 mid-trimester inductions of abortion at our teaching hospital was reviewed. Termination of pregnancy was carried out with 1mg of vaginal gemeprost every 3h up to three doses over a 24-h period, repeated the following day if necessary. Failed induction was defined as women undelivered by 96 h. The study population was then stratified by gestational age, parity, gravidity and previous uterine scars. Main outcome parameters were failed induction and complication rates. Statistical analysis was performed using the chi(2) test or Fisher's exact test for categorical data, and the t-test and linear regression for continuous variables. RESULTS: No significant differences were found in the primary outcome parameters with regard to the obstetric parameters considered. The failed induction rate was 1.2% with an overall incidence of complications of 7.4%. Parity was the main factor that affected clinical response (time to abortion interval and number of pessaries). CONCLUSION: Patients' obstetric history does affect the clinical response to gemeprost, but its safety and effectiveness are preserved. These data provide clinicians with important information for correct counselling. PMID- 16309823 TI - European Urology is "Your" Journal. PMID- 16309824 TI - Retinoic acid amplifies the host immune response to LPS through increased T lymphocytes number and LPS binding protein expression. AB - Vitamin A deficiency is associated with increased susceptibility to infection but the effects of Vitamin A supplementation on host response to pathogens are controversial. This study investigated the mechanisms by which all-trans retinoic acid (atRA) modulates the host immune response in an experimental model of Vitamin A supplementation before and after challenge with LPS in rats. We show here that a supplementation with five daily injections of 10mg/kg atRA increased the number of T lymphocytes in the peripheral blood. In addition, we show that atRA increased the expression of the LPS binding protein (LBP), a component of the LPS recognition system. The retinoic acid receptor (RAR)alpha agonist Ro 4060 55 but not the pan-retinoid X receptors (RXRs) agonist Ro 2573-86 mimicked the effects of atRA on LBP expression suggesting that atRA enhances LBP expression through a RARalpha-mediated pathway. In order to investigate the significance of increased LBP expression we challenged atRA-supplemented rats with the Gram positive bacteria Listeria monocytogenes (LM) that activates the immune response independently from LBP. In sharp contrast to our previous observations that atRA supplementation enhances IFN-gamma expression and NOS2 pathway activation in LPS challenged rats [Devaux, Y., Grosjean, S., Seguin, C., David, C., Dousset, B., Zannad, F., Meistelman, C., de Talance, N., Mertes, P.M., Ungureanu-Longrois, D., 2000. Retinoic acid and host-pathogen interactions: effects on inducible nitric oxide synthase in vivo. Am. J. Physiol. 279, E1045-E1053], atRA did not increase the LM-induced IFN-gamma expression and NOS2 pathway activation. Overall, these data demonstrate that although atRA induces a "priming" of the immune system characterized by increased T lymphocytes number and LBP expression, the profile of the immune response depends on the inflammatory/infectious stimulus. These results could explain why Vitamin A supplementation could have beneficial/neutral or deleterious effects according to the identity of the infectious pathogen. PMID- 16309825 TI - Molecular modeling of new camptothecin analogues to circumvent ABCG2-mediated drug resistance in cancer. AB - Irinotecan (CPT-11) is a widely used potent antitumor drug that inhibits mammalian DNA topoisomerase I (Topo I). However, overexpression of ABCG2 (BCRP/MXR/ABCP) reportedly confers cancer cells resistance to SN-38, the active form of CPT-11. To circumvent the ABCG2-associated drug resistance, the structure activity-relationship (SAR) of 14 new camptothecin (CPT) analogues has been studied with respect to the substrate specificity of ABCG2. While the lactone E ring is a prerequisite for anticancer activity, modifications of the A or B rings do not significantly affect Topo I inhibition. Based on the substrate specificity of ABCG2, it is strongly suggested that CPT analogues with a hydroxyl group at position 10 or 11 of the A ring are recognized by ABCG2 and are thereby effectively extruded from cancer cells. To develop a platform for the molecular modeling to circumvent anticancer drug resistance, we have carried out quantum chemical calculations and neural network SAR analysis. Electrostatic potential iso-surfaces generated by ab initio MO calculations using restricted Hartree-Fock method have revealed that negative potential localized at positions 10 or 11 in the A ring is important for recognition by ABCG2. PMID- 16309826 TI - Increase of disintergin metalloprotease 10 (ADAM10) expression in oral squamous cell carcinoma. AB - The A disintergin and metalloprotease (ADAM) superfamilies play important roles in angiogenesis, development, and tumorigenesis. Amyloid precursor protein (APP) is an important protein related to Alzheimer's disease. Recent research shows that ADAM10 alpha-secretase activity can release the secreted form of APP. We have previously demonstrated an increase of APP expression in oral squamous cell carcinoma (OSCC) and related OSCC cell lines. The present study characterizes ADAM10 expression in the neoplastic process of OSCC. RT-PCR analysis revealed a two-fold increase in APP mRNA expression in 50% of OSCC (n=50) relative to corresponding non-malignant matched tissues (NMMT). This increase in mRNA expression occurred at the preneoplastic stage. A significant correlation between mRNA expression of ADAM10 and APP in OSCC was noted. A non-buccal subset of OSCC correlated with an increase of mRNA expression of both ADAM10 and APP. The increase of ADAM10 protein expression in the majority of OSCC tissues and cell lines studies was confirmed by Western blot analysis. Additionally, an increase of ADAM10 immunoreactivity in OSCC relative to NMMT was noted. An antisense oligonucleotide against ADAM10 reduced ADAM10 expression as well as growth in an OSCC cell line. However, this treatment did not reduce the secreted form APP. This study suggests that ADAM10 expression plays a role in the carcinogenesis of OSCC and proliferation of OSCC cells, independent of APP processing. PMID- 16309827 TI - Enhanced TiO2 photocatalytic degradation of bisphenol E by beta-cyclodextrin in suspended solutions. AB - Enhancement of beta-cyclodextrin (beta-CD) on TiO(2) photocatalytic degradation of bisphenol E (BPE, bis(4-hydroxyphenyl)ethane) was investigated under a 250 W metal halide lamp (lambda> or =365 nm) in this work. In the system of photocatalytic degradation of BPE, the photodegradation rate of BPE in aqueous solutions containing beta-CD and TiO(2) was obviously faster than that in aqueous solutions containing only TiO(2). After 40 min of irradiation, beta-CD could increase the photodegradation efficiency by about 26% for 10 mg l(-1) BPE in the UV-vis/TiO(2) system and the photodegradation of 2.5-20.0 mg l(-1) BPE in aqueous solutions was found to follow pseudo-first-order law and the adsorption constant and the reaction rate constant of BPE in the system containing beta-CD and TiO(2) are obviously higher than those in the system containing only TiO(2), the influence factors on photodegradation of BPE were studied and described in details, such as beta-CD concentration, pH, BPE initial concentration and gas medium. The formation of CO(2) as a result of mineralization of BPE was observed during the photodegradation process. After 120 min of irradiation, the mineralization efficiency of BPE reached 61% in the presence of beta-CD, whereas mineralization efficiency was only 23% in the absence of beta-CD. The enhancement of photodegradation of BPE could be dependent on the enhancement of adsorption of BPE on TiO(2) surface and moderate inclusion-depth of BPE in the beta-CD cavity. PMID- 16309828 TI - Utilization of water clarifier sludge for copper removal in a liquid fluidized bed reactor. AB - A method was used to investigate the potential for using water clarifier sludge to remove copper in a fluidized-bed reactor (FBR). This study was conducted to evaluate the removal of copper in an aquatic system without prior treatment. Chemical analyses of water clarifier sludge through inductively coupled plasma atomic emission spectrophotometry indicated that silicon, aluminum and iron oxides made up more than 84% of this average composition, similar to the composition of clay. The experimental results indicated that the copper removal efficiency was highly dependent on the pH. pH values also influence the character of the water clarifier sludge. After the copper was adsorbed by the water clarifier sludge, the pH of the solution was slightly increased. In the FBR, the copper removal efficiency reached 90% when the initial copper concentration was 20 mg/L, the pH was 4 and the operating time was 60 min. In addition, copper precipitation occured on the surface of clarifier sludge when the initial copper concentration was 20 mg/L and the pH was 7.0. The kinetics of copper ion adsorption at pH 5 showed that 94% copper ion was removed in 60 min and the adsorption equilibrium was attained in 5 h. The mechanisms of adsorption of copper ions on water clarifier sludge including the formation of surface complexation and surface precipitate. PMID- 16309829 TI - Evaluation of lower flammability limits of fuel-air-diluent mixtures using calculated adiabatic flame temperatures. AB - The lower flammability limit (LFL) of a fuel is the minimum composition in air over which a flame can propagate. Calculated adiabatic flame temperatures (CAFT) are a powerful tool to estimate the LFL of gas mixtures. Different CAFT values are used for the estimation of LFL. SuperChems is used by industry to perform flammability calculations under different initial conditions which depends on the selection of a threshold temperature. In this work, the CAFT at the LFL is suggested for mixtures of fuel-air and fuel-air-diluents. These CAFT can be used as the threshold values in SuperChems to calculate the LFL. This paper discusses an approach to evaluate the LFL in the presence of diluents such as N2 and CO2 by an algebraic method and by the application of SuperChems using CAFT as the basis of the calculations. The CAFT for different paraffinic and unsaturated hydrocarbons are presented as well as an average value per family of chemicals. PMID- 16309830 TI - Cystatin C expression is increased in the hippocampus following photothrombotic stroke in rat. AB - Stroke is a major cause of epilepsy, but the molecular mechanisms underlying post stroke epileptogenesis are unknown. The expression of cystatin C, a cysteine protease inhibitor, is increased in the hippocampus during status epilepticus (SE)-induced epileptogenesis, and regulates both cell death and birth. To test the hypothesis that increased cystatin C expression represents a common molecular alteration induced by epileptogenic brain insults, we investigated the time course, cellular localization, and association of cystatin C expression with neuronal damage during post-stroke epileptogenesis. Stroke was induced with photothrombosis, which leads to epilepsy in approximately 20-30% of rats. Cystatin C expression was increased in the CA1 area of the hippocampus 4 days after photothrombosis, when the diameter of the lesion was the largest. Double labeling and confocal analysis indicated that cystatin C was expressed in astrocytes and microglia. Unlike after SE, cystatin C expression did not change in the dentate gyrus. Also, increased cystatin C expression was not associated with neurodegeneration, which was demonstrated as an absence of Fluoro Jade B positive cells in adjacent sections. The present study provides evidence that cystatin C may be involved in cellular alterations that occur after an epileptogenic insult, not only after SE but also after photothrombotic stroke. PMID- 16309831 TI - Changes of parvalbumin immunoreactive neurons and GFAP immunoreactive astrocytes in the rat lateral geniculate nucleus following monocular enucleation. AB - The expression of calcium binding proteins (CaBPs) has been linked to protection of neurons. The present study investigated the effects of monocular enucleation on the distribution of parvalbumin immunoreactive (PV-IR) neurons and glial fibrillary acidic protein immunoreactive (GFAP-IR) astrocytes in both the dorsal (dLGN) and ventral (vLGN) regions of the lateral geniculate nucleus (LGN). Our results demonstrated an increase in PV-IR neuronal density on the contralateral vLGN at 1-week post-enucleation (PE), which was maintained without significant change for 12 weeks. By contrast, PV-IR neurons in dLGN decreased significantly at all time point examined. The number of GFAP-IR astrocytes showed an initial increase from 1 to 4 weeks PE and then gradually decreased until 48 weeks in both regions of the LGN with contralateral side predominance. The present results suggest that monocular enucleation results in variable expression of PV-IR neurons and GFAP-IR astrocytes in the LGN complex, which may play an important role in neuronal degeneration and neuroplasticity of the rat visual system. PMID- 16309832 TI - The cyclooxygenase 2 -765 C promoter allele is a protective factor for Alzheimer's disease. AB - The cyclooxygenase-2 enzyme (COX-2) is of particular importance in the inflammatory response and recent findings have demonstrated a considerable role in Alzheimer's disease (AD) pathogenesis. In order to assess the possible putative role of a COX-2 polymorphism (765G/C) in AD, we examined its distribution in 161 community-based controls and 168 AD clinic-based cases previously recruited from memory disorder clinics in Tampa and Miami, Florida. There were no significant differences between the two groups in age/age of onset or gender. A significant difference was observed in the distribution of the COX-2 -765 alleles between AD cases and controls (chi(2) = 6.565, p = .010; OR = .596; CI = [.401-.888], p = .011), with the frequency of the C allele being higher in controls. In addition, a significant difference was observed for this polymorphism by genotype (chi(2) = 6.561, p = .038) and by presence or absence of C+ genotypes (chi(2) = 6.207, p = .013; OR = .464, CI = [.351-.885], p = .013). In this sample, the C allele of COX-2 -765 promoter polymorphism is associated with decreased risk of Alzheimer's disease, a finding which further supports the involvement of COX-2 in AD etiology. PMID- 16309833 TI - Mitochondrial damage induced by fetal hyperphenylalaninemia in the rat brain and liver: its prevention by melatonin, Vitamin E, and Vitamin C. AB - Abnormal oxidative stress was observed in hyperphenylalaninemia and other inborn errors of intermediary metabolism, owing to the accumulation of toxic metabolites, free radical production and increased LPO products. In our model of maternal hyperphenylalaninemia, pregnant rats were injected with 300 mg/kg BW l phenylalanine (PHE) and 50 mg/kg BW p-chlorophenylalanine (PCPA) dissolved in saline. In this research study, we measured LPO-by-products, i.e., malonaldehyde (MDA) and 4-hydroxynonenal (4-HNE) and we demonstrated that maternal hyperphenylalaninemia increased both markers of oxidative stress in the brain and liver mitochondria of the pups. We also demonstrated that administration of melatonin, Vitamin E, and Vitamin C, in this order of potency, prevented the oxidative damage to the mitochondria, especially in the brain. We therefore conclude that maternal hyperphenylalaninemia induces a clear state of oxidative stress that is somehow directly involved in brain and liver impairment, which can be prevented by melatonin, Vitamin E, and Vitamin C. PMID- 16309834 TI - Structural and neurochemical comparison of vagal and spinal afferent neurons projecting to the rat lung. AB - Afferent information from the lung is conveyed both to the brainstem and to the spinal cord by primary afferent fibres originating from vagal sensory (jugular nodose ganglion complex=JNC) and dorsal root ganglion (DRG) neurons, respectively. Most interest, so far, has been paid to the vagal pathway while much less is known about spinal afferents. Here we provide the first direct comparison of rat pulmonary spinal and vagal pulmonary afferent neurons with respect to structural (soma size) and two neurochemical characteristics (binding of lectin IB4, immunoreactivity to calcitonin gene-related peptide=CGRP). After retrograde labelling from the lung, all possible combinations of CGRP immunoreactivity and IB4-binding were observed, and the neurochemically defined subpopulations occurred in the same order of frequency in DRG and JNC: (1) IB4( )/CGRP(+) (DRG: 48%, JNC: 47%); (2) IB4(-)/CGRP(-) (DRG: 35%, JNC: 29%); (3) IB4(+)/CGRP(+) (DRG: 12%, JNC: 21%) and (4) IB4(+)/CGRP(-) (DRG: 5%, JNC: 3%). In the IB4(-)/CGRP(-) population, pulmonary DRG neurons were slightly, but significantly larger than those in JNC (mean diameter: 33 microm versus 30 microm). This group is likely to contain slowly and rapidly adapting mechanoreceptors, which may be differently distributed among rat vagal and spinal afferent pathways. In rat DRG, labelling patterns IB4(-)/CGRP(+), IB4(+)/CGRP(+) and IB4(+)/CGRP(-) are generally characteristic for different nociceptor subtypes. With respect to these features and soma size, no further distinction between spinal and vagal afferents became obvious, although this does not exclude elicitation of entirely different responses when these pathways are stimulated. PMID- 16309835 TI - Aquaporin 1 immunoreactive enteric neurons in the rat ileum. AB - Most neurons in the central nervous system and peripheral nervous system do not express water transporting protein, aquaporin (AQP). In the present study, we have demonstrated the presence of AQP1 immunoreactivity in a particular neuronal subtype in the enteric nervous system (ENS) of the rat ileum. AQP1-immunoreactive (IR) neurons simultaneously expressed a neuronal marker HuC/D. Moderate numbers of AQP1-IR neuronal somata were found in the myenteric plexus, and a very few were found in the submucosal plexus. AQP1-IR neurons can be classified as Dogiel type I cells, which have several short processes and a single long process. Many AQP1-IR fibers were found both in the myenteric and submucosal plexi. Many AQP1 IR varicose fibers were closely associated with neuronal somata in the ganglia, whereas other AQP1-IR fibers penetrated into the muscle layers. These results suggest that AQP1-IR neurons probably play a significant role within the ENS to control gut functions. PMID- 16309836 TI - Electrophysiologically identified subpopulations of taste bud cells. AB - The heterogeneous population of mammalian taste cells includes several cellular subtypes specializing in distinct physiological functions. They are poorly understood at the single cell level because the available physiological data have generally been obtained from unidentified taste cells. We recorded them from individual taste cells isolated from circumvallate, foliate, and fungiform papilla of the mouse, employing the patch clamp technique, and tried to elucidate whether universal electrophysiological criteria may be established for the identification of functionally different cellular subpopulations. It was found that irrespective of the papillae type, most ( approximately 96%) of robust taste cells could be categorized into three distinct subgroups on the basis of families of whole-cell (WC) currents exhibited in response to membrane polarization. The validity of this quite simple criterion was further confirmed by using different voltage clamp protocols, ion substitutions, and channel blockers to record different ionic currents, including voltage-gated (VG) Ca(2+), inward-rectifying K(+), and hyperpolarization-activated currents. Given that our findings are based on the statistically significant number of recordings, we believe that the electrophysiological identification of taste cells presented here may be effective for further studies on single taste cell physiology, including taste transduction. PMID- 16309837 TI - A late wake time phase delays the human dim light melatonin rhythm. AB - Short sleep/dark durations, due to late bedtimes or early wake times or both, are common in modern society. We have previously shown that a series of days with a late bedtime phase delays the human dim light melatonin rhythm, as compared to a series of days with an early bedtime, despite a fixed wake time. Here we compared the effect of an early versus late wake time with a fixed bedtime on the human dim light melatonin rhythm. Fourteen healthy subjects experienced 2 weeks of short 6h nights with an early wake time fixed at their habitual weekday wake time and 2 weeks of long 9 h nights with a wake time that occurred 3h later than the early wake time, in counterbalanced order. We found that after 2 weeks with the late wake time, the dim light melatonin onset delayed by 2.4 h and the dim light melatonin offset delayed by 2.6 h (both p < 0.001), as compared to after 2 weeks with the early wake time. These results highlight the substantial influence that wake time, likely via the associated morning light exposure, has on the timing of the human circadian clock. Furthermore, the results suggest that when people truncate their sleep by waking early their circadian clocks phase advance and when people wake late their circadian clocks phase delay. PMID- 16309838 TI - On the importance of fifth-order spherical aberration for a fully corrected electron microscope. AB - Next generation aberration correctors will not only eliminate the third-order spherical aberration, but also improve the information limit by correction of chromatic aberration. As a result of these improvements, higher order aberrations, which have largely been neglected in image analysis, will become important. In this paper, we concern ourselves with situations where sub-A resolution can be achieved, and where the third-order spherical aberration is corrected and the fifth-order spherical aberration is measurable. We derive formulae to explore the maximum value of the fifth-order spherical aberration for directly interpretable imaging and discuss the optimum imaging conditions and their applicable range. PMID- 16309839 TI - Probing the population of the spin-orbit split levels in the actinide 5f states. AB - Spin-orbit interaction in the 5f states is believed to strongly influence exotic behaviors observed in actinide metals and compounds. Understanding these interactions and how they relate to the actinide series is of considerable importance. To address this issue, the branching ratio of the white-line peaks of the N4,5 edge for the light actinide metals, alpha-Th, alpha-U, and alpha-Pu were recorded using electron energy-loss spectroscopy (EELS) in a transmission electron microscope (TEM) and synchrotron-radiation-based X-ray absorption spectroscopy (XAS). Using the spin-orbit sum rule and the branching ratios from both experimental spectra and many-electron atomic spectral calculations, accurate values of the spin-orbit interaction, and thus the relative occupation of the j = 5/2 and 7/2 levels, are determined for the actinide 5f states. Results show that the spin-orbit sum rule works very well with both EELS and XAS spectra, needing little or no correction. This is important, since the high spatial resolution of a TEM can be used to overcome the problems of single-crystal growth often encountered with actinide metals, allowing acquisition of EELS spectra, and subsequent spin-orbit analysis, from nm-sized regions. The relative occupation numbers obtained by our method have been compared with recent theoretical results and they show a good agreement in their trend. PMID- 16309840 TI - Experimental inoculation of beagle dogs with Ehrlichia species detected from Ixodes ovatus. AB - Three beagle dogs were inoculated with mice spleen/liver homogenate infected with Ehrlichia species detected from Ixodes ovatus (EIO) and one dog was used as a control. All three infected dogs did not show clinical signs of disease except for mild pyrexia throughout the 41-day study period. Splenomegaly was observed from Day 7 post-inoculation (p.i.) in two of the dogs. Hematological and biochemical abnormalities included mild thrombocytopenia, hypoproteinaemia, hypoalbuminaemia and increased C-reactive protein values. One of the dogs' splenic aspirate sample was PCR-positive for Ehrlichia Day 7 p.i. and another dogs' blood and bone marrow aspirate sample was PCR-positive Day 41 p.i. Sequence analysis of the PCR products showed 100% homology with the 16SrRNA partial gene sequence of Ehrlichia sp. HF565. Antibody titers to EIO were observed in all three experimentally infected dogs starting from the first week p.i. and cross reactivity with Ehrlichia canis was detectable in one of the dogs starting Day 7 p.i. These data suggest that infection of dogs with EIO is possible, though is probably of low pathogenic importance. Cross-reactivity of EIO infected dog serum with E. canis raises the likelihood of false E. canis seropositive dogs. PMID- 16309841 TI - Prescription-only anthelmintics--a questionnaire survey of strategies for surveillance and control of equine strongyles in Denmark. AB - In 1999, legislation in Denmark made anthelmintic drugs available only by prescription, and prohibited their use for routine, prophylactic treatment. A questionnaire survey was conducted in 2004 to determine current strategies for surveillance and control of equine strongyles used in Danish equine veterinary practices. Eighty-seven of 170 (51.2%) registered equine veterinary practices responded. Ninety seven percent of the respondents used faecal egg counts for diagnosis and surveillance, and 41% used larval cultures. Logistic regression revealed that the use of larval cultures was positively correlated with numbers of employees (P = 0.013) and the proportion of equine caseload in the practice (P < 0.000). Performing faecal egg counts and treating horses was most frequent during spring, summer and early autumn. Veterinary practices made treatment decisions based on cut-off values ranging from 20 to 500 eggs per gram (EPG) faeces. Ages of horses, clinical suspicions of parasitic disease, or requests by the owners were the most important factors in the strategies for faecal sampling. Less commonly, sampling strategies included all horses on the premises and random sampling. Ninety five percent of the respondents reported treating certain groups of horses without prior faecal analysis, including horses with clinical signs of parasitic disease (77%), foals (84%), horses less than 3 years of age (52%), and pregnant mares (51%). The respondents regarded the cyathostomin group as the most prevalent cause of parasitic disease and ill-thrift, followed by large strongyles and Parascaris equorum. Sixty seven percent rotated regularly between drugs, while 11% performed routine screening for anthelmintic resistance. Results of this survey suggest that limiting equine anthelmintics to prescription-only availability has increased the level of strongyle surveillance. Veterinary practitioners play a central role in equine parasite management with indications of a lowered intensity of treatment. However, screening for anthelmintic resistance remains uncommon. PMID- 16309842 TI - Vagal and splanchnic afferent nerves are not essential for anorexia associated with abomasal parasitism in sheep. AB - Heavy burdens of the abomasal nematode, Ostertagia (Telodorsagia) circumcincta, in growing lambs result in a reduction in liveweight gain due largely to a drop in voluntary feed intake. The present study investigated: (1) the role of subdiaphragmatic vagal and non-vagal visceral afferent nerves in mediating a reduction in voluntary feed intake, using subdiaphragmatic vagal deafferentation (vagotomy) either alone or in combination with coeliac-superior mesenteric ganglionectomy (vagotomy and sympathectomy); and (2) the association between appetite, abomasal pH, selected blood values (amidated gastrin (G-17-amide), glycine-extended gastrin (G-17-Gly), pepsinogen and leptin) and worm burden, in sheep experimentally infected with 100,000 O. circumcincta infective larvae per os. Neither vagotomy alone nor vagotomy and sympathectomy in combination adversely affected the establishment or course of development of the parasite burden, when compared with a control group subject to sham surgery. Furthermore, neither surgical procedure prevented the drop in appetite seen 5-10 days post infection, although combined vagotomy and sympathectomy did reduce voluntary feed intake prior to the start of the study. Ostertagia infection resulted in a significant increase in abomasal pH in all three groups, which was accompanied by an increase in blood G-17-amide and in G-17-Gly, the latter reported for the first time in parasitized ruminants. There were no significant differences in blood leptin, also reported for the first time in parasitized sheep, either between groups or in comparison with pre-infection levels, though weak negative correlations were established between blood leptin and appetite from day 5 to the end of the study in all three groups and a positive correlation with blood G-17 amide in the control group over the same period. These data suggest that neither intact subdiaphragmatic vagal afferent nerves or coeliac-superior mesenteric ganglion fibres, nor changes in circulating gastrin and leptin concentrations play a major role in mediating the hypophagic effects of O. circumcincta in parasitized sheep. PMID- 16309843 TI - A modified critical test and its use in two dose titration trials to assess efficacy of praziquantel for Anoplocephala perfoliata in equids. AB - Aims of this study with 43 equids naturally infected with Anoplocephala perfoliata in two dose titration trials were to document (i) the usefulness of a critical test with a 48 h treatment to necropsy period, (ii) efficacy of an oral paste of 0.5-2.0 mg praziquantel/kg body weight, and (iii) when after treatment would fecal egg counts provide best estimates of the tapeworm's prevalence in a herd. All feces passed by an equid after treatment and collected in successive 12 h batches were examined for tapeworms. At necropsy, tapeworms were identified as attached to the mucosa or unattached. Tapeworms were examined with a stereoscope and identified as normal or abnormal. Fecal samples were taken for egg counts at treatment and at 6 h intervals thereafter. In 32 of 36 treated equids, efficacy was 100% and mean efficacies for 0.5, 1.0, 1.5 and 2.0 mg praziquantel/kg body weight were 85.5, 99.7, 100 and 100%, respectively. Two equids treated with 0.5 mg praziquantel/kg body weight had low efficacies (5.4 and 38.1%) and normal tapeworms were found attached in the ventral colon and in one equid also in the dorsal colon. In seven untreated equids, tapeworms were primarily in the cecum with 21.3% detached. "Major fragments" or worms without a scolex but otherwise nearly a complete worm were 20.5% of the number of intact worms; they were not included in the efficacy analysis but should be. If the two equids with low efficacy were eliminated and if the number of all tapeworms and major fragments are combined less than 0.5% were in feces within 12 h of treatment, about 20% were in the 12-24 h period, 42% in 24-36h, 24% in 36-48 h and 13.5% in the equids at necropsy. One horse passed all its tapeworms in 24 h. This 48 h test when compared with a 24 h one with no examination of feces was more efficient in use of trial animals and reduced underestimation and overestimation of efficacy. However, a protocol similar to the 48 h test but with a 24 or 36 h post-treatment period should be investigated. The mean egg count peaked 18-24 h after treatment and samples taken at that time would provide the best estimate of prevalence. The Cornell-Wisconsin centrifugal flotation technique had a specificity of 100% and at 18 and 24 h its sensitivity was 94%. A brief discussion on critical and controlled tests for assessing efficacy of an anthelmintic for A. perfoliata is presented. PMID- 16309844 TI - Assessment of anti-estrogenic activity of tamoxifen in transgenic mice expressing an enhanced green fluorescent protein gene regulated by estrogen response element. AB - Tamoxifen is an anti-estrogenic agent for the treatment of breast cancer, while exhibiting estrogenic activity in such tissues as the uterus. This study aimed to test whether these opposite properties of tamoxifen in the uterus can be evaluated separately in vivo. We employed two transgenic murine models named, respectively, the ERE-EGFP Ar+/+ mouse and ERE-EGFP Ar-/- mouse. Both types of mice possess an enhanced green fluorescent protein (EGFP) gene regulated by four copies of estrogen response elements (EREs), while the latter lacks a functional aromatase gene, which encodes an enzyme catalyzing conversion of androgens to estrogens. Tamoxifen clearly exhibited estrogenic activity in the uteri of ERE EGFP Ar-/- mice, as it caused uterine wet weight gain and E2-target gene induction, as 17beta-estradiol (E2) did. However, tamoxifen did not enhance the EGFP expression in ERE-EGFP Ar-/- mice, although E2 induced it significantly. In ERE-EGFP Ar+/+ mice, tamoxifen suppressed the EGFP expression in a time- and dose dependent manner. Thus, the present study demonstrated that estrogenic and anti estrogenic activities of tamoxifen can be evaluated by using ERE-EGFP Ar-/- and ERE-EGFP Ar+/+ mice, respectively. Furthermore, these animal models are useful to select and evaluate estrogenic and anti-estrogenic activities of chemical compounds. PMID- 16309845 TI - Kinetic analysis of the inhibition of human butyrylcholinesterase with cymserine. AB - Accompanying the gradual rise in the average age of the population of most industrialized countries is a regrettable progressive rise in the number of individuals afflicted with age-related neurodegenerative disorders, epitomized by Alzheimer's disease (AD) but, additionally, including Parkinson's disease (PD) and stroke. The primary therapeutic strategy, to date, involves the use of cholinesterases inhibitors (ChEIs) to amplify residual cholinergic activity. The enzyme, acetylcholinesterase (AChE), along with other elements of the cholinergic system is depleted in the AD brain. In contrast, however, its sister enzyme, butyrylcholinesterase (BuChE), that likewise cleaves acetylcholine (ACh), is elevated and both AChE and BuChE co-localize in high amounts with the classical pathological hallmarks of AD. The mismatch between increased brain BuChE and depleted levels of both ACh and AChE, particularly late in the disease, has supported the design and development of new ChEIs with a preference for BuChE; exemplified by the novel agent, cymserine, whose binding kinetics are characterized for the first time. Specifically, as assessed by the Ellman method, cymserine demonstrated potent concentration-dependent binding with human BuChE. The IC50 was determined as 63 to 100 nM at the substrate concentration range of 25 to 800 microM BuSCh. In addition, the following new binding constants were investigated for human BuChE inhibition by cymserine: T(FPnubeta), K(nubeta), K(Bs), K(MIBA), M(IC50), D(Sc), R(f), (O)K(m), OIC100, K(sl), theta(max) and R(i). These new kinetic constants may open new avenues for the kinetic study of the inhibition of a broad array of other enzymes by a wide variety of inhibitors. In synopsis, cymserine proved to be a potent inhibitor of human BuChE in comparison to its structural analogue, phenserine. PMID- 16309846 TI - The modulatory effects of nicotine on parietal cortex activity in a cued target detection task depend on cue reliability. AB - This functional magnetic resonance imaging study investigates the effects of nicotine in a cued target detection task when changing cue reliability. Fifteen non-smoking volunteers were studied under placebo and nicotine (Nicorette polacrilex gum 1 and 2 mg). Validly and invalidly cued trials were arranged in blocks with high, middle and low cue reliability. Two effects of nicotine were investigated: its influence on i) parietal cortex activity underlying the processing of invalid vs. valid trials (i.e. validity effect) and ii) neural activity in the context of low, middle and high informative value of the cue (i.e. cue reliability effect). Nicotine did not affect behavioral performance. However, nicotine reduced the difference in the blood oxygenation level dependent (BOLD) signal between invalid and valid trials in the right intraparietal sulcus. The reduction of parietal activity in invalid trials was smaller in the low cue reliability condition. The same posterior parietal region exhibited a nicotinic modulation of BOLD activity in valid trials which was dependent on cue reliability: Nicotine specifically enhanced the neural activity during valid trials in the context of low cue reliability, i.e. when subjects are already in a state of low certainty. We speculate that the right intraparietal sulcus might be part of two networks working in parallel: one responsible for reorienting attention and the other for the cholinergic modulation of cue reliability. By reducing the use of the cue, nicotine modulates parietal activity related to reorienting attention in conditions with higher cue certainty. On the other hand, nicotine increases parietal activity in states of low certainty. This enhanced activation might influence brain regions, such as the posterior cingulate, directly involved in the processing of cue reliability. PMID- 16309847 TI - Molecular heterogeneity along the dorsal-ventral axis of the murine hippocampal CA1 field: a microarray analysis of gene expression. AB - There has been increasing interest in functional heterogeneity along the septotemporal, dorsal-ventral (D-V) axis of the hippocampus. Although anatomical connectivity and lesion studies point to discrete roles for these sub-regions, the contribution of differential gene expression across this axis has not been systematically studied. Here we present findings from an Affymetrix microarray screen aimed at identifying genes in the CA1 region of the adult murine hippocampus that show significant differential expression along the D-V axis. Our results indicate that the vast majority of monitored genes (>90%) had tissue expression levels that differed by less than 20% between regions, while less than 0.1% of genes had expression levels that varied more than three-fold by sub region. Only 23 probes showed a CA1 dorsoventral signal intensity ratio greater than three: 18 enriched dorsally and five enriched ventrally. Probes with the greatest difference in expression levels represent a range of genes with known functions in patterning and signaling, as well as genes without known function. Selective screening with digoxigenin-labeled in situ hybridization confirms the existence of CA1 sub-regionalized expression, with some genes exhibiting a graded expression pattern across the D-V axis, and others restricted to a discrete region. Our findings demonstrate that there are gene expression differences across the D-V axis of the adult murine hippocampus within traditionally recognized cytoarchitecturally defined boundaries. Combined with the previously recognized differences in connectivity and results from lesion studies, our data further confirm the existence of functional heterogeneity along the D-V axis. PMID- 16309848 TI - Methamphetamine use among young adults: health and social consequences. AB - The current research analyzed the relationship between methamphetamine use and health and social outcomes. Interviews were conducted with a sample of 106 respondents. Virtually all of the respondents experienced negative consequences of methamphetamine use. The most serious, but least prevalent, methamphetamine related health problem was seizures and convulsions. The most prevalent health effect was weight lose. A substantial number of respondents experienced severe psychological symptoms: depression, hallucinations, and paranoia. Of the 106 respondents, 34.9% had committed violence while under the influence of methamphetamine. The data suggest that methamphetamine-based violence was more likely to occur within private domestic contexts, both family and acquaintance relationships. It is apparent from the findings that methamphetamine use heightens the risk for negative health, psychological, and social outcomes. Having said this, it is crucial to acknowledge that there was no evidence of a single, uniform career path that all chronic methamphetamine users follow. Furthermore, a significant number of sample members experienced limited or no serious social, psychological, or physical dysfunction as a result of their methamphetamine use. PMID- 16309849 TI - Induction of heat shock proteins may combat insulin resistance. AB - The molecular mechanism responsible for obesity-associated insulin resistance has been partially clarified: increased fatty acid levels in muscle fibers promote diacylglycerol synthesis, which activates certain isoforms of protein kinase C (PKC). This in turn triggers a kinase cascade which activates both IkappaB kinase beta (IKK-beta) and c-Jun N-terminal kinase (JNK), each of which can phosphorylate a key serine residue in IRS-1, rendering it a poor substrate for the activated insulin receptor. Heat shock proteins Hsp27 and Hsp72 have the potential to prevent the activation of IKK-beta and JNK, respectively; this suggests that induction of heat shock proteins may blunt the adverse impact of fat overexposure on insulin function. Indeed, bimoclomol--a heat shock protein co inducer being developed for treatment of diabetic neuropathy--and lipoic acid- suspected to be a heat shock protein inducer--have each demonstrated favorable effects on the insulin sensitivity of obese rodents, and parenteral lipoic acid is reported to improve the insulin sensitivity of type 2 diabetics. Moreover, there is reason to believe that heat shock protein induction may have a favorable impact on the microvascular complications of diabetes, and on the increased risk for macrovascular disease associated with diabetes and insulin resistance syndrome. Heat shock protein induction may also have potential for preventing or treating neurodegenerative disorders, controlling inflammation, and possibly even slowing the aging process. The possible complementarity of bimoclomol and lipoic acid for heat shock protein induction should be assessed, and further efforts to identify well-tolerated agents active in this regard are warranted. PMID- 16309850 TI - cGMP may have trophic effects on beta cell function comparable to those of cAMP, implying a role for high-dose biotin in prevention/treatment of diabetes. AB - Incretin hormones have trophic effects on beta cell function that can aid prevention and treatment of diabetes. cAMP is the primary mediator of these effects, and has been shown to potentiate glucose-stimulated insulin secretion, promote proper beta cells differentiation by increasing expression of the crucial transcription factor PDX-1, and prevent beta cell apoptosis. cGMP's role in beta cell function has received far less scrutiny, but there is emerging evidence that it may have a trophic impact on beta cell function analogous to that of cAMP. An increase in plasma glucose boosts beta cell production of cGMP, which acts as a feed-forward mediator to enhance glucose-stimulated insulin secretion. cGMP also has an anti-apoptotic effect in beta cells, and there is now indirect evidence that it promotes expression of PDX-1. Supraphysiological concentrations of biotin can directly activate guanylate cyclase, and there is limited evidence that high intakes of this vitamin can be therapeutically beneficial in diabetics and in rodent models of diabetes. Beneficial effects of cGMP on muscle insulin sensitivity and on control of hepatic glucose output may contribute to biotin's utility in diabetes. The fact that nitric oxide/cGMP exert a range of favorable effects on vascular health should further encourage exploration of biotin's preventive and therapeutic potential. If an appropriate high-dose biotin regimen could achieve a modest systemic increase in guanylate cyclase activity, without entailing unacceptable side effects or risks, such a regimen might have considerable potential for promoting vascular health and preventing or managing diabetes. PMID- 16309851 TI - The possible role of tissue-type plasminogen activator and the plasminogen system in the pathogenesis of major depression. AB - Major depressive disorder (MDD) is one of the most common psychiatric illnesses with an unknown etiology. Evidence from animal and human studies has suggested that brain-derived neurotrophic factor (BDNF) function may be implicated in the pathogenesis of MDD. Tissue-type plasminogen activator (tPA) is a highly specific serine proteinase that catalyses the generation of zymogen plasminogen from the proteinase plasmin. Recent studies have found that the proteolytic cleavage of proBDNF, a BDNF precursor, to BDNF by the plasmin represents a mechanism by which the direction of BDNF action is controlled. Furthermore, studies using mice deficient in tPA has demonstrated that tPA is important for the stress reaction, a common precipitating factor for MDD. A study of the serum levels of the plasminogen activator inhibitor-1 (PAI-1), the major inhibitor of tPA, found that women with MDD had a higher PAI-1 concentration than normal controls. From these findings, it is proposed that the tPA/plasminogen system may play a role in the pathogenesis of MDD. Attempts to confirm the tPA/plasminogen hypothesis may lead to new directions in the study of the pathogenesis of MDD and the development of a novel intervention of this disorder. PMID- 16309852 TI - Disease-specific survival following the brachytherapy management of prostate cancer. AB - PURPOSE: To determine disease-specific survival (DSS) and associated predictive factors after prostate brachytherapy. METHODS AND MATERIALS: A total of 1561 patients underwent brachytherapy for prostate cancer from 1990 to 2004 (median follow-up, 3.8 years). Treatment included brachytherapy alone (n = 634), brachytherapy and hormonal therapy (n = 420), and implant and external beam therapy (n = 507). RESULTS: The DSS and overall survival rates at 10 years were 96% and 74%, respectively. Gleason score significantly impacted DSS, with 10-year rates of 98%, 91%, and 92% for scores of < or = 6, 7, and > or = 8, respectively (p < 0.0001). Multivariate analysis revealed that PSA status after treatment had the most significant effect on DSS. Ten-year DSS rates were 100%, 52%, and 98%, respectively for patients without PSA failure (n = 1430), failure with a doubling time (DT) < or = 10 months (n = 64), and failure with a DT > 10 months (n = 67), respectively (p < 0.0001). In patients with PSA failure, DSS rates were 30%, 67%, and 98%, for those with DT < or = 6 months, > 6-10 months, and > 10 months, respectively (p < 0.0001). CONCLUSIONS: The 10-year DSS rate supports the efficacy of brachytherapy. Patients dying with disease within 10 years after treatment harbor inherently aggressive cancer with high Gleason scores and short DT. PMID- 16309853 TI - [Nerve growth factor (NGF): new role for a neuronal mediator in the airways]. PMID- 16309854 TI - Interrupting timing in interval production and discrimination: similarities and differences. AB - Interruptions in human timing have been studied in the last few years using temporal production and discrimination tasks. Expecting a break shortened perceived duration in both paradigms but manipulating break duration affected time production only, suggesting that preparatory processes might not take place in time discrimination. In time production, using cues revealed that providing information about the break may modulate the effect of break expectancy. For example, time was perceived as shorter when a break was expected in trials with no breaks, but forewarning participants of the break absence with a cue almost abolished the effect. In the present experiment, a tone was classified as "short" or "long" in a discrimination task. Location and duration of breaks were varied and cues were provided in some trials with no breaks. Results showed an effect of break expectancy: perceived duration shortened with increasing pre-break duration. Reducing expectancy with cues in uninterrupted tones decreased the proportion of "short" responses in long-tone trials, but not in short-tone trials. As in previous discrimination experiments, perceived duration was unaffected by varying break duration. Similarities and differences in results as well as in their interpretation when breaks are used in time production and time discrimination tasks are discussed. PMID- 16309855 TI - A truncated acidic domain in Xenopus TRF1. AB - Telomere function is mediated by a complex of proteins bound to double-stranded and single-stranded telomeric repeats. A key player in this complex is TRF1, which binds to duplex TTAGGG repeats and acts as a negative regulator of telomere length. This protein's domain structure, as defined by studies with mammalian orthologs, consists of an N-terminal acidic domain, a dimerization domain, and a C-terminal Myb DNA binding domain. TRF1 from Xenopus laevis was cloned and sequenced, and the encoded protein found to have a similar structure but with a very short acidic domain. This short acidic domain was confirmed in Xenopus tropicalis, a true diploid, by cloning of cDNA sequences by RACE and analysis of the genomic locus. The TRF1 transcript is expressed in developing and adult frogs. Compared to the mammalian orthologs, the Xenopus genes are the most distantly related vertebrate examples characterized to date. Since adult Xenopus ubiquitously express somatic telomerase activity, proteins that regulate telomerase access to the chromosome ends are important in regulating telomere length in normal somatic tissue. The structure of Xenopus TRF1 has implications for its regulation by tankyrase. PMID- 16309856 TI - The galectin-3 gene promoter binding proteins in the liver of rats 48-h post treatment with CCl4. AB - The present study was undertaken to characterize structure-function relationships of the rat galectin-3 gene promoter especially focusing on the promoter binding proteins included in livers injured with CCl4. Transcription start site determination identified a 66-nucleotide-long exon 1 of this gene. Transient expression analysis using a reporter luciferase gene assigned a region between 161 and -15 to the proximal promoter within the 1-kb region flanking the 5'-end of exon 1. The rat galectin-3 gene promoter possesses a Runx2 binding site and inverted repeats of Sp1 binding motifs in separate regions downstream from -117 as structures resembling those of the mouse galectin-3 gene promoter. The -161/ 118 region bound two different proteins. One is a novel protein, a rat version of Purbeta that binds to a guanine nucleotide pair at -145 and -144 to modulate constitutive galectin-3 gene transcription. Southwestern blot analysis using the 161/-118 ligand revealed a signal of a 50-kDa protein in liver nuclear extracts from rats 48-h post-treatment with CCl4, but not in those from Ac2F cells and normal rat livers. The inducible nature of this protein suggested its distinctive role in galectin-3 induction in a liver injured with CCl4. E-box and peroxisome proliferator response element-like motifs reside on separate DNA strands from 140 to -135. Contribution of this segment to the regulation of galectin-3 gene transcription under pathological conditions was suggested, since a DNA ligand with the two motifs simultaneously mutagenized at -136 and -137 was not bound by the 50-kDa protein. PMID- 16309857 TI - Identification of functional elements in the murine Gabp alpha/ATP synthase coupling factor 6 bi-directional promoter. AB - The GA-repeat binding protein (GABP) is a ubiquitous transcription factor involved in transcriptional regulation of genes encoding proteins involved in a variety of cellular processes including adipocyte differentiation, mitochondrial respiration, and neuromuscular signaling. GABP is composed of two subunits; the GABP alpha subunit is a member of the Ets-family of transcription factors, and the unrelated ankyrin repeat containing GABP beta subunit. We previously identified a bidirectional promoter directing the expression of Gabpa (GAA) gene in one direction and ATP Synthase Coupling Factor 6 (Atp5j) (CF6) gene in the other [Chinenov, Y., Coombs, C. and Martin, M. E., 2000a. "Isolation of a bi directional promoter directing the expression of the mouse GABP alpha and ATP Synthase Coupling Factor 6 genes. Gene 261:311-320.]. In this study we characterize sequence elements and regulatory factors contributing to the promoter activities of the GAA/CF6 bidirectional promoter. The core of the GAA/CF6 bidirectional promoter is retained within a 400 bp sequence and contains four GABP binding sites, a Sp1/3 binding site and an YY1 binding site. Site directed mutagenesis demonstrated that while no single factor binding site was essential for promoter activity in either direction, the GA1 site located proximal to the previously mapped transcription start sites functioned cooperatively with the other GABP binding sites and with the Sp1/3 and YY1 sites to provide transcriptional activation of the GAA and CF6 promoters. The other GABP sites and the Sp1/3 and YY1 binding sites were functionally redundant for basal promoter activities in both directions. Electrophoretic mobility shift assays identified multiple DNA-protein complexes containing GABP alpha, GABP beta, Sp1, Sp3 or YY1 proteins, including one ternary complex containing GABP alpha, GABP beta and Sp1 proteins. Binding of GABP to the GAA/CF6 bi-directional promoter provides the potential for autoregulation of GABP alpha expression and confirms the importance of GABP in the coordinate expression of respiratory chain components. PMID- 16309858 TI - Mycoflora of poultry feeds and ochratoxin-producing ability of isolated Aspergillus and Penicillium species. AB - In Brazil, commercial feedstuffs are an important component in modern animal husbandry, but there is no information available about fungal contamination and ochratoxin A (OTA) production. The aims of this study were to determine the mycoflora incidence in poultry feeds and evaluate OTA production. In addition, the ability to produce OTA by several Aspergillus and Penicillium species was investigated. A total of 96 samples of poultry feeds were collected from four factories in Rio de Janeiro. Samples were examined for total moulds, for Aspergillus and Penicillium spp. occurrence and for their relative densities on dichloran rose bengal chloramphenicol and dichloran 18% glycerol media. The capacity to produce ochratoxin A by selected Aspergillus and Penicillium species was determined by HPLC. Total mould counts were generally higher than 1 x 10(5 )CFU ml(-1). Aspergillus and Penicillium species were isolated in the highest numbers. Aspergillus flovus and Penicillium citrinum were the most prevalent species. There was a high percentage of potential OTA producers (46%). The amount of OTA produced on this substrate was enough to cause adverse effects in animals. Several strains isolated from poultry feeds were able to produce high levels of OTA on chloramphenicol yeast medium. OTA in raw materials needs to be surveyed and storage practices must be investigated to determine occurrence and establish the livestock toxicological risk in poultry feed. PMID- 16309859 TI - A mechanism for organophosphate-induced delayed neuropathy. AB - Neuropathy target esterase (NTE) reacts with those organophosphates, which cause paralysis with swelling and degeneration of distal parts of long nerves in the legs and spinal cord. Cloning of NTE cDNA allowed the generation of constitutive and brain-specific NTE-null mice: the former die by mid-gestation whereas the latter display age-dependent neurodegeneration. NTE is not required by dividing cells but is needed for survival of post-mitotic cells such as placental secondary giant cells and brain neurons. NTE is localised to the cytoplasmic face of the endoplasmic reticulum (ER) and catalyses the deacylation of ER-membrane phosphatidylcholine (PtdCho) to soluble products-glycerophosphocholine and fatty acids. PtdCho is the major phospholipid of eukaryotic cell membranes. Yeast mutants lacking NTE are viable because they maintain membrane homeostasis by reducing the rate of PtdCho synthesis. By contrast, brain neurons and glia in Drosophila NTE-null mutants accumulate PtdCho, have abnormal membrane structures, and finally undergo cell death. In the nervous system of susceptible vertebrates, neuropathic organophosphates will cause a transient loss of NTE activity, putatively disrupting membrane phospholipid homeostasis and ER functions including axonal transport and glial-axonal interaction: the distal parts of long axons will be particularly vulnerable to loss of these support functions. PMID- 16309860 TI - Short-term delayed-release microcapsules spraycoated with acrylic terpolymers. AB - A series of poly(ethyl acrylate (EA)/methyl methacrylate (MMA)/2-hydroxyethyl methacrylate (HEMA)) lattices were synthesized to prepare short-term delayed release microcapsules by employing the Wurster coating process. Latex with a HEMA molar fraction exceeding 60% could not be synthesized as an aqueous suspension due to latex particle precipitation. The effects of monomer composition on the particle size of latex and the water-uptake and glass transition temperature (T(g)) of cast films were investigated. Lattices whose T(g) ranged from 40 to 80 degrees C were used to prepare the microcapsules. Most of the lattices exhibited excellent process performance while coating particles that were smaller than 100 microm: the product yields were 85.1-90.6% and the mean particle sizes were 82-85 microm. However, since the lattices with high molar ratios of EA and HEMA were highly hydrophilic and strongly adhesive, the core particles in the coating were severely agglomerated. The microcapsules coated with lattices whose HEMA molar fractions were higher than 50% were unable to retard the release of carbazochrome sodium sulfonate, a water-soluble model drug, during the initial 0.5 min. Poly(EA/MMA/HEMA) with a molar ratio of 9:9:10 appeared to be suitable for the preparation of short-term delayed-release microcapsules by the Wurster coating process. PMID- 16309861 TI - AIF and endoG translocation in noise exposure induced hair cell death. AB - Activation of caspases is a key element in the apoptotic process. However, mitochondria also play an important role via the release of proapoptotic proteins. This study investigated the roles of mitochondria-related apoptosis inducing factor (AIF) and endonuclease G (endoG), mitochondrion-specific nucleases, as well as caspase-3, an important mediator of apoptosis, in noise exposure induced hair cell death. Guinea pigs were exposed for 4h/day to broadband noise at 122 dB SPL for 2 days. After the noise exposure, the cochleae were examined for the activity of caspase-3 with carboxyfluorescein-labeled fluoromethyl ketone (FMK)-peptide inhibitors. The cochleae were further examined for AIF and endoG translocation from the mitochondria by immunohistochemistry. Noise exposure triggered activation of caspase-3 in apoptotic hair cells. In the normal organ of Corti, AIF and endoG were co-localized to the mitochondria. After noise exposure, AIF translocated into the nuclei of apoptotic and necrotic hair cells. The translocation of endoG from mitochondria into the nucleus was also found in apoptotic OHCs. These findings indicate that mitochondria-released proapoptotic proteins, AIF and endoG, are important factors in a noise-induced hair cell death pathway. PMID- 16309863 TI - Interaction in the perceptual processing of interaural time and level differences. AB - Phillips and Hall [Psychophysical evidence for adaptation of central auditory processors for interaural differences in time and level, Hear. Res., 202 (2005) 188-199.] recently described the frequency-specific, selective adaptation of perceptual channels for interaural differences in level (ILD) and time (ITD). Psychometric functions for laterality based on ITD or ILD were obtained before and after exposure to adaptor tones of two frequencies presented alternately and highly lateralized to opposite sides. Following adaptation, points of perceived centrality (PPCs) were displaced towards the sides of the adaptor tones, and in opposite directions for the two frequencies. That is, laterality judgements showed a shift away from the adapted side, particularly for test cue values near the middle of the range. These data were congruent with a two-channel, opponent process model of sound laterality coding. The present study used the same general paradigm to explore the independence of perceptual ITD and ILD processing. Psychometric functions for laterality based on ITD or ILD were obtained for each of two frequencies concurrently, before and after exposure to adaptor tones lateralized using the complementary cue. Once again, PPCs derived from the psychometric functions were displaced towards the sides of the adaptor tones, consistent with an opponent-process account of sound laterality coding. The size of the adaptation effect was at least as great as that described in the earlier study. Thus, a quarter cycle ITD adapting stimulus effected a 3 dB shift in the mean ILD-based PPC, and a 12 dB ILD adapting stimulus effected a 100 micros shift in the mean ITD-based PPC. These data offer new evidence concerning interaction in the processing of ITDs and ILDs. PMID- 16309862 TI - Characterization of hearing loss in aged type II diabetics. AB - Presbycusis - age-related hearing loss - is the number one communicative disorder and a significant chronic medical condition of the aged. Little is known about how type II diabetes, another prevalent age-related medical condition, and presbycusis interact. The present investigation aimed to comprehensively characterize the nature of hearing impairment in aged type II diabetics. Hearing tests measuring both peripheral (cochlea) and central (brainstem and cortex) auditory processing were utilized. The majority of differences between the hearing abilities of the aged diabetics and their age-matched controls were found in measures of inner ear function. For example, large differences were found in pure-tone audiograms, wideband noise and speech reception thresholds, and otoacoustic emissions. The greatest deficits tended to be at low frequencies. In addition, there was a strong tendency for diabetes to affect the right ear more than the left. One possible interpretation is that as one develops presbycusis, the right ear advantage is lost, and this decline is accelerated by diabetes. In contrast, auditory processing tests that measure both peripheral and central processing showed fewer declines between the elderly diabetics and the control group. Consequences of elevated blood sugar levels as possible underlying physiological mechanisms for the hearing loss are discussed. PMID- 16309864 TI - WITHDRAWN: The effect of erythrodiol-3-acetate on the expression of matrix metalloproteinase-9 caused by ultraviolet-irradiated cultured human keratinocyte cells. AB - Ahead of Print article withdrawn by publisher. PMID- 16309865 TI - In vitro effect of medicinal plants used to treat erectile dysfunction on smooth muscle relaxation and human sperm. AB - Chloroform and ethanol extracts of root bark of Securidaca longepedunculata, Wrightia natalensis and Rhoicissus tridentata were investigated for their in vitro activity on the contraction of corpus cavernosal smooth muscle of white New Zealand rabbits. Some of the extracts of these plants relaxed the corpus cavernosal smooth muscle at low concentrations. The highest activity was obtained from Securidaca longepedunculata chloroform extracts at a concentration of 13.0 mg/ml, which induced 66.6% relaxation. Viagra was used as a positive control in this study. Extracts of Securidaca longepedunculata added to human spermatozoa affected certain sperm parameters negatively at 6.5 mg/ml and higher whilst there was no effect at 1.0 mg/ml. PMID- 16309866 TI - X-chromosome STR polymorphism of Luoba Ethnic Group living in Tibet (SW China). AB - Allele frequencies of 10 X-chromosome STR loci, namely DXS7133, DXS6789, DXS6804, DXS6799, DXS8378, DXS101, DXS7424, DXS7132, DXS7423 and HPRTB were determined from healthy unrelated males and female individuals of Luoba Ethnic Group living in Tibet (SW China). PMID- 16309867 TI - Pleomorphic adenoma of the external auditory canal complicated by hearing loss secondary to chronic otitis media. AB - Pleomorphic adenoma (PA) rarely occurs in the external auditory canal (EAC). A case of PA complicated with chronic otitis media is reported. The patient was a 36-year-old male who began to experience a hearing loss in the left ear in 1996. He sought medical attention only after ear pain developed in April 2003. At the initial examination, a tumor covered by smooth skin was seen filling the left EAC. Pure-tone audiometry showed residual hearing at low frequencies and scale out across middle to high frequency range. The CT and MRI findings led to a diagnosis of a tumor of the left EAC complicated with an inflammatory middle-ear lesion. The tumor was excised en bloc with the overlying skin, and tympanoplasty was performed. The pathological diagnosis was PA of the left EAC and inflammatory granuloma of the middle ear. PMID- 16309868 TI - Model experiments of BPPV using isolated utricle and posterior semicircular canal. AB - OBJECTIVES: This study was aimed to experimentally investigate the effect of returned otoconia on the utricular using isolated utricles. The effect of interposed otoconia in models of canalolithiasis and cupulolithiasis were also investigated using isolated posterior semicircular canal (PSC). METHODS: Bullfrogs were used. The utricles (Experiment I) and PSC (Experiment II) were removed in Ringer solution. Experiment I-a: The otoconia were carefully removed from the utricular macula with gentle flush of Ringer solution. Before and after the otoconial removal, sinusoidal rotatory stimulation (0.1 Hz, 135 degrees ) was given to record utricular compound action potentials (CAPs). Experiment I-b: (1) Instantaneous changes in the utricular potentials when the otoconial mass was positioned on the macula were recorded. (2) Utricular CAP changes in response to sinusoidal rotation immediately and 10 min after the otoconial positioning were recorded. Experiment II: PSC CAPs due to sinusoidal rotatory stimulation in normal specimen, canalolithiasis and cupulolithiasis models were recorded. RESULTS: Experiment I-a: The utricular CAPs in response to sinusoidal rotation showed sinusoidal oscillation. However, this oscillation disappeared after the otoconial removal. Experiment I-b: (1) The utricular potentials transiently increased for 3-4 s after positioning the otoconial mass. (2) The utricular CAPs increased in seven specimens and decreased in four. Ten minutes after the CAPs were almost the same as immediately after otoconial positioning. Experiment II: In cupulolithiasis model, the PSC CAPs decreased in all specimens. CONCLUSIONS: The otoconia played an essential role as a transducer of acceleration to the utricular macula. Otoconia returned to the utricular macula change utricular reactivity and hence are the possible cause of dizziness after physical therapy. PSC responses to sinusoidal rotation were suppressed in cupulolithiasis model. PMID- 16309869 TI - [Cervical spondylodiscitis: one pathogen may hide another]. AB - A 54-year-old man presented with tuberculous spondylodiscitis associated to E. coli found in an intervertebral disc space needle biopsy. The enterobacteria came from a cholecystitis. The patient was cured by medical treatment, consisting in a non-surgical immobilization, antitubercular quadritherapy in association with a specific antibiotic treatment. No other case of spondylodiscitis caused by a mycobacterial coinfection pathogen has been reported so far. PMID- 16309870 TI - [Pseudomonas aeruginosa isolated in immunocompromised patients: antimicrobial resistance, serotyping, and molecular typing]. AB - OBJECTIVE: Our study dealt with antibiotic resistance and serotypes of Pseudomonas aeruginosa strains isolated from immunocompromised patients in the National Bone Marrow Transplant Center of Tunis as well as molecular typing of ceftazidime resistant strains (CAZ-R). DESIGN: We studied a total of 87 non replicate P. aeruginosa isolates from 36 patients (84 strains) or the hospital environment (3 strains). RESULTS: Rates of antimicrobial resistance were 36% for ceftazidime, 16% for imipenem, 38% for amikacin, and 57% for ciprofloxacin. The 31 CAZ-R strains were associated with O:11 serotype in 84% of the cases. Genetically characterization of CAZ-R strains by Pulsed Field Gel Electrophoresis (PFGE) after digestion of genomic DNA with SpeI revealed 2 genotypic groups. The first was composed of strains isolated from one outpatient between November 1998 and April 1999. Resistance phenotypes of these strains varied after use of antimicrobial drugs. The second was predominant (18/31 CAZ-R strains) in both hematology and graft units and persisted from June 1998 to June 2000 among 5/8 patients. These strains had O:11 serotype in 78% of the cases. The strains of this group were not isolated on patient admission and were isolated from 2 washbasins in the graft unit in May 1999. CONCLUSION: These results suggest the spread of multidrug-resistant O:11 P. aeruginosa clone from a tap water among hospitalized patients in our center, emphasizing the need of standard control of washbasins to eradicate this reservoir. PMID- 16309871 TI - [Surveillance of animal and human rabies in Lithuania from 1991 to 2001]. AB - OBJECTIVE: The objective of this study was to assess the epidemiology of rabies in Lithuania from 1991 to 2000. MATERIALS AND METHODS: Data available from the reports of the Lithuanian State Food and Veterinary Service, of the Public Health centers, of the Veterinary Laboratories of counties, districts and cities, and from the reports of the Lithuanian Ministry of Agriculture and the Environment Protection Department were used to assess rabies both in domestic animals and wildlife. The situation of human rabies from 1991 to 2000 was evaluated using the medical records of patients with a diagnosis of rabies and the epidemiological follow-up of people receiving post-exposure treatment. RESULTS: From 1991 to 2000, 2,148 laboratories confirmed cases of rabies were diagnosed in animals in Lithuania. Domestic animals accounted for 44% of all cases whereas wild animals accounted for 56% of the cases. During the 10-year period of investigation, 5 human cases were reported and 44,710 bitten persons received post-exposure treatment. COMMENTS: A high incidence of rabies in wild animals in Lithuania had a direct influence on the increase human rabies infection. It is necessary to improve preventive measures against rabies - vaccination of domestic and wild animals, and spread information on rabies to the general public. PMID- 16309872 TI - Eight cases of fascioliasis: clinical and microbiological features. AB - Fascioliasis is a parasitic infection caused by the ingestion of food contaminated with Fasciola hepatica. OBJECTIVE AND DESIGN: We report an epidemic of eight cases of fascioliasis in southern France, and describe the clinical features, and the diagnostic and therapeutic tools. RESULTS: Our series includes almost every clinical form described, apart from the pseudotumoral form. Early diagnosis relied mainly on serum assays. Among these, counter-electrophoresis was the first to be positive and therefore appears to be the most useful test, including cases where patients are asymptomatic or mildly ill. Patients were treated with the usual dose of triclabendazole, but six of them required a double dose to recover. CONCLUSION: The use of 20 mg/kg triclabendazole is safe and efficient for patients with acute fascioliasis when a single dose has failed to cure them. PMID- 16309873 TI - [Emergent pathogens, international surveillance and international health regulations (2005)]. AB - In order to address the vitality of the microbial world, to detect emerging infectious diseases, to determine their potential threat to public health, and to establish effective interventions, the World Health Organization (WHO) has developed and coordinates the Global Outbreak Alert and Response Network (GOARN) which connects several surveillance networks. Some of these networks are specific to epidemic-prone diseases, such as influenza, dengue, yellow fever or meningitis. Others were especially designed to track unusual events--such as the emergence of SARS--that are naturally-occurring, accidental, or deliberately created (biological weapons, bio-terrorism). Lastly, a special effort is being made at the international level to modernize the International Health Regulations, now obsolete, and to support all the countries in the reinforcement of their outbreak alert and response capacity. PMID- 16309874 TI - [Evolution of severe pediatric malaria in Togo between 2000 and 2002]. AB - The aim of this study was to analyse the clinical and evolutive aspects of severe malaria in hospitalised children in 2000, 2001, and 2002 in Togo. The study included 361 children in the pediatrics department of Lome-Tokoin University hospital. All them received a 10% dextrose infusion, then an infusion of quinine or intramuscular artemether. Malaria accounted for 4.37% of all hospitalizations. Children aged 1 to 5 years were more affected (69.53%). The most frequent clinical forms were anaemia (55.7%) followed by cerebral manifestations. The frequency of hemoglobinuria increased (17.2%) as well as renal failure (3%) compared to previous years. Thirty-five children died (9.7%). Most of them presented with anaemia, neurological manifestations, or respiratory distress. Neurological sequels were present in 2.2% of patients. PMID- 16309875 TI - [Streptococcus pyogenes: a rare etiology of non-gonococcal urethritis and balanitis in the adult]. AB - Urethritis and balanitis due to S. pyogenes are very uncommon. The authors report the case of a 22 year-old male patient, presenting with this association, the first case reported in France. There was no associated STD. PMID- 16309876 TI - [Vertebral and spinal cord hydatidosis: contribution of imaging]. AB - The authors report the case of a vertebromedullary hydatidosis revealed by medullar compression in a 40-year-old male patient. Magnetic resonance imaging showed lesions of the seventh and eighth dorsal vertebras, cystic lesions in the epidural space and in the perivertebral soft tissues. Surgical excision lead to clinical cure. Vertebromedullary hydatidosis is rare and severe. Modern techniques of imaging are very helpful for the diagnosis and the follow-up of patients after treatment. PMID- 16309877 TI - High-resolution respirometry--a modern tool in aging research. AB - Alterations in mitochondrial function are believed to play a major role in aging processes in many species, including fungi and animals, and increased oxidative stress is considered a major consequence of altered mitochondrial function. In support of this theory, a lot of correlative evidence has been collected, suggesting that changes in mitochondrial DNA accumulate with age in certain tissues. Furthermore, genetic experiments from lower eukaryotic model organisms, indicate a strong correlative link between increased resistance to oxidative stress and an extended lifespan; in addition, limited experimental evidence suggests that the inhibition of mitochondrial function by selected pharmacologically active compounds can extend lifespan in certain species. However, changes in mitochondrial function may affect aging in a different way in various tissues, and a clear statement about the role of mitochondrial deterioration during physiological aging is missing for most if not all species. At this point, respirometric analyses of mitochondrial function provide a tool to study age-associated changes in mitochondrial respiratory chain function and mitochondrial ATP production within living cells and isolated mitochondria. In the recent years, new instruments have been developed, which allow for an unprecedented high-resolution respirometry, which enables us to determine many parameters of mitochondrial function in routine assays using small samples of biological material. It is conceivable that this technology will become an important tool for all those, who are interested in experimentally addressing the mitochondrial theory of aging. In this article, we provide a synopsis of traditional respirometry and the advances of modern high-resolution respirometry, and discuss how future applications of this technology to recently established experimental models in aging research may provide exciting new insights into the role of mitochondria in the aging process. PMID- 16309878 TI - In vitro quantitative determination of ophthalmic irritancy by the chorioallantoic membrane test with trypan blue staining as alternative to eye irritation test. AB - The damage provoked by some substances on the chicken egg's chorioallantoic membrane (CAM) is used as an alternative assay to determine ocular irritation. There is good prediction of the eye irritation when compared to the in vivo Draize method. Nevertheless, this assay has some limitations, such as subjectivity. Hagino et al. developed an objective evaluation technique using the amount of trypan blue absorbed at the site of treatment as an indicator of injury to the CAM. The present work was aimed at the determination of ocular irritation of 21 substances (chemicals and cosmetics). We used the spectrophotometric quantification by trypan blue staining of the damage produced on CAM, of fertile chicken eggs. Results were compared with the values obtained by the traditional Draize assay. We observed a good correlation (r=0.835; p<0.0001) between the amount of dye absorbed by the CAM and the Draize eye irritation test score. The r values were 0.688; p<0.05 for cosmetics and 0.925; p<0.0001 for chemicals. Three chemicals turned as false positive and one cosmetic substance as false negative. The CAM-TBS assay is inexpensive, simple and provides an in vitro alternative method to predict the damage that chemical substances or cosmetics can cause to the ocular structures. PMID- 16309879 TI - Organotypic human vaginal-ectocervical tissue model for irritation studies of spermicides, microbicides, and feminine-care products. AB - A three-dimensional organotypic vaginal-ectocervical (VEC) tissue model has been developed to test the irritation of topically applied spermicides, microbicides, and vaginal-care products. The in vitro tissue model was reconstructed using normal VEC epithelial cells and is well stratified, containing differentiated basal, suprabasal, intermediate, and superficial cell layers similar to in vivo tissue. The intermediate and superficial cell layers contain glycogen, and the expression of cytokeratins 13 and 14 in the tissue also parallels that of native tissue. The MTT viability assay and histological assessment were used to test inter-lot and intra-lot reproducibility. The MTT average intra-lot coefficient of variation (CV) was less than 10% and the time required to reduce tissue viability by 50% (ET-50) following application of 1% Triton X-100 averaged 1.25+/-0.24h (n=23) upon completion of the 11-day culture period and 1.30 h+/- 0.19 for the same tissues stored overnight at 4 degrees C on agarose gels. The utility of the VEC model for irritation studies was examined by testing commercially available products using the MTT assay and histological assessment. The average ET-50 values ranged between 1.8 and 2.7h for feminine washes, 3.9-6.7 h for spermicides, 6.8-18 h for anti-itch creams, and >18 h for douches, lubricants, and anti-fungal creams. Studies of cytokines released from VEC cultures following product application showed that elevated concentrations of IL-1alpha and IL-1beta were associated with toxicity of test materials. In conclusion, the VEC tissue model is a highly reproducible, non-animal means to assess the irritation of contraceptives, microbicides, and vaginal-care products. PMID- 16309880 TI - Defective repair of radiation-induced DNA damage is complemented by a CHORI-230 65K18 BAC clone on rat chromosome 4. AB - The Long Evans cinnamon (LEC) rat is highly susceptible to X-irradiation due to defective DNA repair and is thus a model for hepatocellular carcinogenesis. We constructed a bacterial artificial chromosome (BAC) contig of rat chromosome 4 completely covering the region associated with radiation susceptibility. We used transient and stable transfections to demonstrate that defective DNA repair in LEC cells is fully complemented by a 200-kb BAC, CHORI-230-65K18. Further analysis showed that the region associated with radiation susceptibility is located in a 128,543-bp region of 65K18 that includes the known gene Rpn1. However, neither knockdown nor overexpression of Rpn1 indicated that this gene is associated with radiation susceptibility. We also mapped three ESTs (TC523872, TC533727, and CB607546) in the 128,543-bp region, suggesting that 65K18 contains an unknown gene associated with X-ray susceptibility in the LEC rat. PMID- 16309881 TI - High-resolution map and imprinting analysis of the Gtl2-Dnchc1 domain on mouse chromosome 12. AB - The imprinted Dlk1-Dio3 region on mouse chromosome 12 contains six imprinted genes and a number of maternally expressed snoRNAs and miRNAs. Here we present a high-resolution sequence analysis of the 1.1-Mb segment telomeric to Gtl2 in mouse and a homology comparison to the human. Ppp2r5c and Dnchc1 at the telomeric end of the analyzed sequence are biallelically expressed, suggesting that the imprinted domain does not extend beyond the paternally expressed Dio3 gene. RT PCR experiments support the predicted presence of a maternally expressed intergenic transcript(s) encompassing Gtl2, Rian, and Mirg. These maternally expressed genes, and also the intergenic transcript(s), show pronounced expression in the adult mouse brain, whereas the paternally transcribed Dio3 and the nonimprinted Ppp2r5c and Dnchc1 are expressed in different tissues. Hence, tissue-specific coregulation of maternally expressed genes might be an important feature of this domain. PMID- 16309882 TI - Chromosomal inversion discovered in C3H/HeJ mice. AB - Mice of the inbred mouse strain C3H/HeJ have been shown to be homozygous for a chromosomal inversion on Chromosome (Chr) 6. The inversion encompasses about 20% of the chromosome from approximately 73 Mb to approximately 116 Mb. The importance of this finding is that linkage crosses using C3H/HeJ will show no recombination in this region of Chr 6. The inversion has no apparent effect on the phenotype of C3H/HeJ mice and its presence should not affect biological studies; however, use of C3H/HeJ mice for genetic analysis of Chr 6 should be avoided or the results interpreted with the inversion in mind. The inversion has been named In(6)1J (inversion Chr 6, Jackson 1). PMID- 16309883 TI - Release of cytokines by brain endothelial cells: A polarized response to lipopolysaccharide. AB - Brain endothelial cells (BECs) comprise the blood-brain barrier (BBB) and are an active part of the neuroimmune system, responding to and transporting cytokines. BECs also have the ability to secrete neuroimmune substances, including cytokines. A unique feature of the BEC is its polarization, with its luminal (blood-facing) and abluminal (brain-facing) cell membranes differing in their lipid, receptor, and transporter compositions. This polarization could have functional consequences for neuroimmune communication. We postulated (i) that cytokine secretion from the luminal or abluminal membranes could differ under baseline or stimulated conditions and (ii) that an immune challenge from one side of the BBB could result in cytokine release from the other. We used an in vitro BBB model of mouse BECs cultured as monolayers to investigate cytokine secretion into luminal and abluminal chambers. Our major findings in these studies were: (i) the first demonstration that interleukin (IL)-1alpha, IL-10, and granulocyte macrophage colony-stimulating factor are secreted from BECs and confirmation of the secretions of IL-6 and tumor necrosis factor-alpha, (ii) that constitutive and lipopolysaccharide (LPS)-stimulated secretion of cytokines is polarized in favor of luminal secretion, and (iii) that response to neuroimmune stimulation is also polarized as exemplified by the finding that abluminal LPS more robustly induced secretion of IL-6 than did luminal LPS. Overall, these findings support the BBB as an important source of cytokines. Furthermore, the BBB can respond to immune challenges received from one side of the neuroimmune axis by releasing cytokines into the other. PMID- 16309884 TI - Endocrine and immunological correlates of behaviorally identified swim stress resilient and vulnerable rats. AB - Animal models of stress-induced depression have identified a bimodal reactivity to stress, namely 'resilience' and 'vulnerability.' Possible corresponding differences in endocrine and immunological responses between these groups have not been delineated. Male Sprague-Dawley rats were divided into three groups: stress (n=25), confined controls (n=7), and home cage controls (n=7). Stress rats were exposed to 80, 5-s inescapable cold water swim trials (15 degrees C). Twenty four hours later, the stress rats were tested on an instrumental swim escape test (SET) but now they had access to an omnidirectional lever that terminated the stress. Immediately after the SET, trunk blood was collected to assay for serum corticosterone (CORT), and spleens were removed and natural killer cell activity (NKCA) and concanavalin A (CON-A) induced lymphocyte proliferation determined. Subjects in the stress treatment group were divided into distinct 'resilient' and 'vulnerable' categories by a median split for average escape latencies across the last 25 trials of the SET. Stress rats secreted more CORT than controls and vulnerable rats secreted greater levels than resilient rats. NKCA was greatest in control rats, and was decreased in the stress rats although the resilient and the vulnerable groups did not differ. Conversely, CON-A-induced lymphocyte proliferation was greatest in stress rats, vulnerable rats exhibiting more proliferation than resilient rats, but both were greater than both control groups. Stress animals were hypothermic throughout the swim stress procedures but exhibited a stress-induced fever following the initial swim trials. The observed differences may have important predictive and theoretical utility for vulnerable and resilient profiles. PMID- 16309885 TI - Evidence supporting a circadian control of natural killer cell function. AB - Natural killer (NK) cells participate in the immune response against infection and cancer. An emerging body of epidemiological data supports that circadian homeostasis may constitute a factor risk for cancer development. Physiological rhythms under circadian control persist in the absence of light entrainment and ultimately rely on a molecular clock. We have previously shown that NK cell cytolytic activity follows a daily rhythm and that NK cells enriched from light entrained rats present 24-h oscillations of clock genes, cytolytic factors, and cytokines. To investigate whether these oscillations are under a genuine circadian control, we assessed the daily expression of clock genes (Per1, Per2, Clock, and Bmal1), a clock-controlled gene (Dbp), cytolytic factors (granzyme B and perforin), and cytokines (IFN-gamma and TNF-alpha) in NK cells enriched from rats maintained in constant darkness (DD). In addition, we investigated whether the disruption of the NK cell clock by RNA interference (RNAi) affects the expression of cytolytic factors and cytokines. Persistent 24-h oscillations were found in the expression levels of clock genes, cytolytic factors, and cytokines in NK cells enriched from DD rats. In addition, RNAi-mediated Per2 knockdown caused a significant decrease of granzyme B and perforin levels in the rat derived NK cell line RNK16. Taken together, these results provide evidence supporting that NK cell function is under circadian regulation. PMID- 16309886 TI - In guinea pig spermatozoa, the procaine-promoted synchronous acrosome reaction results in highly fertile cells exhibiting normal F-actin distribution. AB - In guinea pig spermatozoa, procaine induces Ca(2+) independent hyperactivated motility suggestive of sperm capacitation. Nonetheless, in the presence of high extracellular Ca(2+), procaine increases cytoplasmic Ca(2+). We analyze the procaine effect on the acrosome reaction (AR) processes in guinea pig spermatozoa. Results indicated that: (i) in spermatozoa pre-incubated 5-30 min in MCM-PLG medium, procaine produced synchronous AR, (ii) the acrosome-reacted sperm number increased with the capacitation period before procaine treatment and with procaine concentration, (iii) acrosome reaction was blocked when Ca(2+) was omitted, (iv) plasma membrane-outer acrosomal membrane fusion started within 2 min after procaine treatment, (v) in acrosome-reacted spermatozoa, actin polymerization occurred and F-actin was located in the equatorial and post acrosomal regions and (vi) procaine treatment resulted in highly fertile acrosome reacted spermatozoa. This is the first report indicating that procaine promotes synchronic AR in mammalian spermatozoa. If procaine promotes premature AR of spermatozoa in vivo, it might be a factor for infertility in patients exposed to this local anesthetic. PMID- 16309887 TI - Classical and modern prejudice: attitudes toward people with intellectual disabilities. AB - In two studies, Study 1 and Study 2, we examine whether attitudes toward people with intellectual disabilities, like sexism and racism, consist of two forms-a classical and a modern, where the classical is overt and blatant and the modern is more subtle and covert. Self-report scales tapping these two forms were developed in Study 1. Based on confirmatory factor analyses, the results in Study 1 supported our hypothesis and revealed that the modern and classical forms are correlated but distinguishable. This outcome was replicated in Study 2. Construct and discriminatory validations of the scales provided further support for the distinction. The theoretical and practical importance of the results is discussed in relation to previous research on attitudes toward people with intellectual disabilities and other social outgroups. PMID- 16309888 TI - Purkinje cell and cerebellar effects following developmental exposure to PCBs and/or MeHg. AB - We recently reported that rats exposed to PCBs and MeHg during development were impaired on the rotating rod, a test of balance and coordination that is often indicative of cerebellar damage. In addition, developmental PCB exposure is known to dramatically reduce circulating thyroid hormone concentrations, which may have a negative impact on cerebellar development. Therefore, we investigated the effects of combined PCB and MeHg exposure on Purkinje cells and the cerebellum. The serum and brains from littermates of the animals tested on the rotating rod were collected at weaning, and we also collected brains from the adult animals at the end of motor testing. Four groups were studied: 1) vehicle controls, 2) PCBs only (Aroclor 1254, 6 mg/kg/d, oral), 3) MeHg only (0.5 ppm, in dams' drinking water), and 4) PCB+MeHg (at the same doses as in individual toxicant exposures). Female Long-Evans rats were exposed beginning 4 weeks prior to breeding with an unexposed male and continuing until postnatal day (PND) 16. There was a significant reduction in serum T4 and T3 concentrations in the PCB and PCB+MeHg pups on PND21. Golgi-impregnated Purkinje cells were examined in PND21 brains, but there were no significant exposure-related effects on primary dendrite length, branching area, or structural abnormalities. However, all three male exposure groups had a marginally significant increase in Purkinje cell height, which may suggest a subtle thyromimetic effect in the cerebellum. Cresyl-violet stained sections from the adult brains showed no exposure-related effects within paramedian lobule in Purkinje cell number, total lobule volume or layer volumes (molecular, granule cell and white matter layers). Evidence is provided for the dysregulation of expression of cerebellar ryanodine receptor (RyR) isoforms in PCB-exposed brains, and this could contribute to the rotating rod deficit by changing critical aspects of intracellular calcium signaling within the cerebellum. PMID- 16309889 TI - The neurotoxic effects of prenatal cardiac glycoside exposure: a hypothesis. AB - Cardiac glycosides (CGs) are beneficial in treating cardiac conditions; depending on time and dosage, they can also be toxic as they regularly cross the blood brain barrier and the placenta and may affect the unborn baby. This paper therefore focuses on the effects of CGs administered to the mother on normal cellular physiology of the foetus with specific reference to neural tissue. CGs act by binding to the Na+/K+-ATPase and decrease or inhibit Na+-K+ pump activity. In the foetus, CGs may disrupt ion homeostasis. An over-dosage of CGs or when it is taken during pregnancy, can also affect the neuro-energy levels of brain tissue in particular. We conclude and hypothesize that CGs in this case will not only cause severe alterations in neuronal function due to disruption of membrane activity, but also in glutamate clearance, affecting neurotransmission in general. Furthermore, elevated cytosolic Ca2+ will lead to permeabilization of the mitochondrial membranes, resulting ultimately in mitochondrial dysfunction. This will result in neurotoxicity--ensuing in neural cell damage or death, and we propose the mechanism to be due to neuro-necrapoptosis. PMID- 16309890 TI - Long-term effects of lanthanum intake on the neurobehavioral development of the rat. AB - The effects of subchronic exposure to lanthanum on rats' physical and neurobehavioral development were investigated. Wistar rats were exposed to lanthanum through oral administration at 0, 0.1, 2, and 40 mg/kg concentrations from gestation day 0 through 5 months of age. Prior to weaning of the pups, physical parameters and neurobehaviors were assessed, including body weight gain, pinna detachment, eye opening, surface righting reflex and swimming endurance. At 30 days of age, DNA concentration and protein/DNA ratio of the whole brain were determined. At 150 days of age, the Morris water maze test was carried out to study the memory and learning abilities of the rats. Differences were found in the body weight gain, surface righting reflex and swimming endurance. Moreover, lanthanum exposure significantly altered the DNA concentration and Protein/DNA in brain. The Morris water maze test showed that lanthanum exposure at 40 mg/kg significantly impaired memory and learning abilities. These findings indicate that lanthanum is a potential behavior teratogen. The information provided by this work should be considered in future applications of lanthanides. PMID- 16309891 TI - Significance of serum CXCL10/IP-10 level in type 1 diabetes. AB - Although we have previously reported an elevated serum level of CXCL10/IP-10 (CXCL10), a Th1 chemokine, in type 1 diabetic patients, little is known about the origin of serum CXCL10 and its significance in type 1 diabetes. Therefore, we examined serum CXCL10 level in NOD mice in association with the expression level of CXCL10 in the pancreas, pancreatic lymph nodes (LN) and spleen. Serum CXCL10 level increased over time towards the onset of diabetes, and was significantly higher in the "diabetic" period (20 and 24 weeks of age and at onset of diabetes) than in the "pre-diabetic" period (4, 8 and 16 weeks of age). Moreover, serum CXCL10 level was associated with CXCL10 and CXCR3 mRNA level in pancreatic LN. Furthermore, it seemed that serum CXCL10 level increased just before (or at) the onset of overt diabetes. These results suggest that serum CXCL10 level may reflect accumulation of Th1 lymphocytes in pancreatic LN, and measurement of serum CXCL10 level may be useful to assess the pathophysiology of the disease course in type 1 diabetes. PMID- 16309892 TI - Measuring pathways to care in first-episode psychosis: a systematic review. AB - BACKGROUND: Adequately understanding and measuring pathways to care is a prerequisite for early detection and effective treatment of first-episode psychosis. METHOD: We conducted a systematic review of studies on pathways to care in first-episode psychosis to establish what measures currently exist to assess pathways in first-episode psychosis and to compare these measures. RESULTS: We identified 15 studies which had used six different measures of pathways to care. Differences in aims, methodology and lack of psychometric data did not allow a direct comparison of pathways measures but certain common themes emerged. DISCUSSION: Pathways to care in first-episode psychosis are diverse and varied. There is no measure with established psychometric properties that has been devised on a well-developed theoretical or conceptual framework and had its psychometric properties established. The conflict between exploring the patient's narrative and journey through the healthcare system and developing an empirical measure of pathways with optimal outcomes has hindered the development of such a measure. PMID- 16309893 TI - Positive family environment predicts improvement in symptoms and social functioning among adolescents at imminent risk for onset of psychosis. AB - This study investigated whether family factors, such as criticism, emotional over involvement (EOI), warmth, and positive remarks, as measured by the Camberwell Family Interview (CFI), predict symptom change and social outcome for individuals identified as at imminent risk for conversion to psychosis. Twenty-six adolescent patients were administered the Structured Interview for Prodromal Syndromes and the Strauss-Carpenter Outcome Scale at baseline and follow-up assessment approximately three months later. Patients' primary caregivers were administered the CFI at baseline. After controlling for symptom severity at baseline, there were significant associations between caregivers' EOI at baseline and improvement in high-risk youths' negative symptoms and social functioning at follow-up. Similarly, caregivers' positive remarks at baseline were associated with improvement in negative and disorganized symptoms at follow-up, and warmth expressed by caregivers was associated with improved social functioning at follow up. Although family members' critical comments were not related to patients' symptoms, the majority of critical remarks were focused on patients' negative symptoms and irritability/aggression, which may be important targets for early intervention. These preliminary results provide a first glimpse into the relationship between family factors and symptom development during the prodrome and suggest that positive family involvement predicts decreased symptoms and enhanced social functioning at this early stage. The finding that four-fifths of the youth enrolled in this early intervention clinical research program have shown symptomatic improvement by the three-month assessment point is very encouraging from an early detection/early intervention standpoint. PMID- 16309894 TI - Antioxidant enzymes and lipid peroxidation in different forms of schizophrenia treated with typical and atypical antipsychotics. AB - There is accumulating evidence of altered antioxidant enzyme activities and increased levels of lipid peroxidation in schizophrenia. Free radical-mediated abnormalities may contribute to specific aspects of schizophrenic symptomatology and complications of its treatment. However, few studies have evaluated both antioxidant enzymes and lipid peroxidation in the same schizophrenic patient groups treated with typical or atypical antipsychotics. Plasma malondialdehyde (MDA) levels and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities were analyzed using established procedures in 92 medicated schizophrenia including paranoid (n=34), disorganized (n=18) and residual subtypes (n=40), as well as in control subjects (n=50). The results showed that activities of SOD and GSH-Px were decreased but levels of MDA were elevated in patients with a chronic form of schizophrenia as compared with normal controls. SOD and GSH-Px activities were found to be significantly lower in paranoid and residual subtypes compared to both disorganized subtype and the control group. MDA levels were significantly higher in all subtypes compared to the control group. There were no significant differences in any parameters measured among all three subgroups treated with clozapine (n=44), risperidone (n=20) and typical antipsychotics (n=28). Additionally, a significantly higher MDA levels, but a significantly lower CAT activity was noted in female than male patients. These results suggest that oxidative stress may be implicated in the pathophysiology of all subtypes of schizophrenia, which may contribute to the increased membrane lipid peroxidation. Long-term treatments with typical and atypical antipsychotics may produce the similar effects on the antioxidant enzymes and lipid peroxidation. PMID- 16309895 TI - Early-onset schizophrenia is associated with impaired adolescent development of attentional capacity using the identical pairs continuous performance test. AB - The authors examined performance on the Continuous Performance Test-Identical Pairs "numbers" task in adolescents with schizophrenia (n=59) and healthy controls (n=55). Adjusting for an estimate of premorbid intelligence and socioeconomic status, patients performed worse than normal controls on all three d' conditions (2-digit, 3-digit, 4-digit). However, there was a significant group by-age-by-condition interaction (F[4,100]=4.69, p<.01) indicating an interaction between development and disease state. At the simplest level of the task (2 digit) the difference between patients with schizophrenia and controls was evident at all ages; while for the more difficult levels of the task (3-digit, 4 digit), differences between groups gradually increased across the tested age span (10 to 20 years of age). Premorbid social isolation was associated with worse attentional performance in patients, suggesting a relationship and continuity with negative symptoms. These data suggest that attentional differences in adolescents with schizophrenia are better captured by different tasks at different ages. The discrepant findings of attentional impairments reported in the literature for adolescents with schizophrenia could reflect the underlying etiological complexity of the disorder that may have a variable impact on involved brain regions and neurocognitive functioning. PMID- 16309896 TI - The association between psychiatrist numbers and hospitalization costs for schizophrenia patients: a population-based study. AB - OBJECTIVE: This study explores the association between psychiatrist case volumes and costs for hospitalized schizophrenia patients. METHODS: The study uses the Taiwan National Health Insurance Research Database for 2003, identifying the study subjects from the database by ICD-9-CM principal diagnosis code 295. Our study sample comprises of 135,621 admissions treated by 787 psychiatrists in 181 hospitals, with the sample being divided equally into three psychiatrist volume groups: or=601 admissions (high volume). After adjusting for psychiatrist, patient and hospital characteristics, multiple regression analyses were performed to determine the association between psychiatrist case volume and hospitalization costs (total, drug, and non-drug). RESULTS: The regression analyses showed that after adjusting for psychiatrist, patient and hospital characteristics, average treatment costs associated with hospitalized schizophrenia patients were inversely related to psychiatrist volume. The respective total costs, drug costs and non-drug costs of patients treated by high-volume psychiatrists were 369 US dollars (p<0.001), 26 US dollars (p<0.001) and 343 US dollars (p<0.001) lower than those of low-volume psychiatrists. The respective total costs, drug costs and non-drug costs for those treated by medium-volume psychiatrists were 248 US dollars (p<0.001), 22 US dollars (p<0.001) and 226 US dollars (p<0.001) lower than those of low-volume psychiatrists. CONCLUSIONS: We find that after adjusting for patient, psychiatrist and hospital characteristics, an inverse volume-cost relationship exists for psychiatrists treating schizophrenia patients. Further studies should aim to investigate the volume-quality relationship to ensure that incremental cost savings associated with increased patient volume are not achieved at the expense of quality of patient care. PMID- 16309897 TI - Characteristic MC1R polymorphism in the Japanese population. PMID- 16309898 TI - Dehydroepiandrosterone (DHEA) attenuates cocaine-seeking behavior in the self administration model in rats. AB - The aim of this study was to determine the possible involvement of the neurosteroid dehydroepiandrosterone (DHEA) in cocaine-seeking behavior in a self administration model in rats. DHEA pretreatment (continued thereafter concomitantly with cocaine self-administration) attenuated cocaine-seeking behavior and elevated the levels of dopamine and serotonin in several brain regions relevant to cocaine addiction. Chronic cocaine self-administration induced elevation in brain DHEA, its sulfate ester, DHEAS, and pregnenolone. The increased brain DHEA following cocaine self-administration may serve as a compensatory protective mechanism geared to attenuate the craving for cocaine. Such anti-craving activity is further enhanced by DHEA treatment before and during cocaine self-administration. PMID- 16309899 TI - Dendritic spines and linear networks. AB - The function of the cortical microcircuitry is still mysterious. Using a bottom up analysis based on the biophysics and connectivity of cortical neurons, we propose the hypothesis that the neocortex is essentially a linear integrator of inputs. Dendritic spines would slow the neuron and contribute to linearize input summation. Since excitatory axons are relatively straight, they appeared designed to help disperse information to a large number of recipient neurons, generating a distributed circuit. A linear summation regime will ensure the full benefit of a distributed connectivity matrix. Linear integration could also help the neocortex decode the sensory world and may have additional computational advantages. In this view, spines would be the anatomical signature of linear networks. PMID- 16309900 TI - [Prevention of childhood asthma: facts, paradox, query]. PMID- 16309901 TI - Determination of carbaryl in natural water samples by a surface plasmon resonance flow-through immunosensor. AB - The analysis of carbaryl in natural water samples was accomplished using a portable immunosensor based on surface plasmon resonance (SPR) technology. The assay was based on a binding inhibition immunoassay format with the analyte derivative covalently immobilized on the sensor surface. An alkanethiol self assembled monolayer (SAM) was formed onto the gold-coated sensor surface to allow the reusability of the same sensing surface during 220 regeneration cycles. Reproducibility was evaluated by performing three independent assays in triplicate on 3 different days. The batch-assay variability was also calculated using three different gold-coated sensor surfaces. The intra- and inter-day relative standard deviation were 8.6 and 15.3%, respectively, whilst a variation of 7.4% in assay sensitivity was obtained by employing different sensor chips. The lowest detection limit, calculated as the concentration providing a 10% decrease of the blank signal, was of 1.38 microg L(-1). Matrix effects were also evaluated in different water types, showing I50 values (carbaryl concentrations that produced a 50% decrease of the blank signal) within the range of carbaryl standard curves in distilled water (2.78-3.55 microg L(-1)). The carbaryl immunoassay performance was validated with respect to conventional high performance liquid chromatography-mass spectrometry (HPLC-MS). The correlation between methods was in good agreement (r2 = 0.998, 0.999 and 0.999) for the three types of natural water samples tested. A complete assay cycle, including regeneration, is accomplished in 20 min. All measurements were carried out with the SPR sensor system (beta-SPR) commercialised by the company SENSIA, SL (Spain). The small size and low-time of response of the beta-SPR platform would allow its utilization in real contaminated locations. PMID- 16309902 TI - Comparative study on chemical pretreatments to accelerate enzymatic hydrolysis of aquatic macrophyte biomass used in water purification processes. AB - In this study, enzymatic hydrolysis of two floating aquatic plants which are suitable for water purification, water hyacinth (Eichhornia crassipes) and water lettuce (Pistia stratiotes L.), was performed to produce sugars. Twenty chemical pretreatments were comparatively examined in order to improve the efficiency of enzymatic hydrolysis. As a result, the alkaline/oxidative (A/O) pretreatment, in which sodium hydroxide and hydrogen peroxide were used, was the most effective pretreatment in terms of improving enzymatic hydrolysis of the leaves of water hyacinth and water lettuce. The amount of reducing sugars in enzymatic hydrolysate of water lettuce leaves was 1.8 times higher than that of water hyacinth leaves, therefore water lettuce seems to be more attractive as a biomass resource than water hyacinth. Although roots of these plants contained large amounts of polysaccharides such as cellulose and hemicellulose, they generated less monosaccharides than from leaves, no matter which chemical pretreatment was tested. PMID- 16309903 TI - Synthesis and antiprotozoal activity of some new synthetic substituted quinoxalines. AB - A series of 29 new quinoxalines was synthesized and evaluated in vitro against several parasites (Leishmania donovani, Trypanosoma brucei brucei, and Trichomonas vaginalis). Several of them displayed interesting activities, and particularly four quinoxaline amides showed in vitro antileishmanial properties (IC50 less than 20 microM). PMID- 16309904 TI - Design, synthesis, and biological evaluation of BODIPY-erythromycin probes for bacterial ribosomes. AB - BODIPY-erythromycin probes of bacterial ribosomes were designed and synthesized by attaching a BODIPY fluorophore to the 4''- and 9-positions of the erythromycin structure. The probes exhibited excellent binding affinity to bacterial ribosomes and competed with erythromycin and other drugs whose binding sites are in the same vicinity of the 50S subunit. The synthetic fluorescent probe 5 was successfully adapted in our ultra high-throughput screening (uHTS) to identify novel ribosome inhibitors. PMID- 16309905 TI - Isolation, synthesis, and anti-tumor activities of a novel class of podocarpic diterpenes. AB - A novel unusual 17-carbon diterpenoid, named (+)-7-deoxynimbidiol, was isolated from the stalks of Celastrus hypoleucus (Oliv.) Warb. Its racemate and derivatives were synthesized, and the inhibitory activities of these compounds against four cultured human-tumor cell lines were evaluated. The structure activity relationship was discussed. PMID- 16309906 TI - Synthesis and pharmacochemical study of novel polyfunctional molecules combining anti-inflammatory, antioxidant, and hypocholesterolemic properties. AB - We have designed and synthesized a series of novel molecules having a residue of a classical NSAID and an antioxidant moiety, both attached through amide bonds to a known nootropic structure, an L-proline, trans-4-hydroxy-L-proline or DL pipecolinic acid residue. The compounds were found to retain anti-inflammatory and antioxidant activities, to acquire hypocholesterolemic action, and to possess a greatly reduced gastrointestinal toxicity. The novel molecules could find useful applications, among others, in slowing the progression or delaying the onset of neurodegenerative diseases. PMID- 16309907 TI - Androstenediol analogs as ER-beta-selective SERMs. AB - A series of 19-substituted androstenediol derivatives was prepared. Some of the novel analogs were surprisingly potent and selective ligands for ER-beta. PMID- 16309908 TI - Microbial transformation of silybin by Trichoderma koningii. AB - Microbial transformation of silybin A (1) and silybin B (2), the major hepatoprotective flavonolignan diastereomers from the fruits of Silybum marianum, with the culture broth of Trichoderma koningii gave two pairs of glucosylated derivatives. Their structures were identified as silybin A 3-O-beta-D glucopyranoside (3), silybin A 7-O-beta-D-glucopyranoside (4), silybin B 3-O-beta D-glucopyranoside (5) and silybin B 7-O-beta-D-glucopyranoside (6) by spectroscopic methods. PMID- 16309909 TI - Structure-activity relationship study of 1,4-dihydropyridine derivatives blocking N-type calcium channels. AB - Cilnidipine is a 1,4-dihydropyridine derived L/N-type calcium channel dual blocker possessing neuroprotective and analgesic effects which are related to its N-type calcium channel inhibitory activity. In order to find specific N-type calcium channel blockers with the least effects on cardiovascular system, we performed structure-activity relationship study on APJ2708, which is a derivative of cilnidipine, and found a promising N-type calcium channel blocker 21b possessing analgesic effect in vivo with a 1600-fold lower activity against L type calcium channels than that of cilnidipine. PMID- 16309910 TI - Permanent patellar dislocation and osteoarthritis of the knee after femoral fracture in childhood. A case report. AB - A 52 year-old woman suffered bilateral femoral shaft fractures when she was 5 years old and they were treated surgically. She had been able to walk without pain and giving way throughout her young and adult life, however, when she came to us with a 1-year history of the right knee pain, she could walk only 10 m continuously. The right knee revealed valgus deformity with complete lateral patellar dislocation. The femoro-tibial angle was 154 degrees and arthritic change was seen in the femorotibial joint. Surfaces of both the patella and the femur were degraded. Twenty-two degrees of rotational deformity was also found internally in the involved side. Varus osteotomy with external rotation of the distal fragment and lateral retinacular release combined with reefing of vastus medialis muscle were performed. Twenty-four months after surgery, knee score and functional score were, respectively, 85/100 and 71/100, using knee society clinical rating system and there was no pain associating patella incongruity. PMID- 16309911 TI - Tricyclic pharmacophore-based molecules as novel integrin alpha(v)beta3 antagonists. Part 1: design and synthesis of a lead compound exhibiting alpha(v)beta3/alpha(IIb)beta3 dual antagonistic activity. AB - In order to generate novel compounds with integrin alpha(v)beta3-antagonistic activity together with antiplatelet activity, tricyclic pharmacophore-based molecules were designed and synthesized. Although piperazine-containing compounds initially prepared were selective alpha(IIb)beta3 antagonists, replacement of piperazine with piperidine furnished a potent alpha(v)beta3/alpha(IIb)beta3 dual antagonist. Structure-activity relationship (SAR) studies provided clues for further development of tricyclic pharmacophore-based integrin antagonists. PMID- 16309912 TI - Tricyclic pharmacophore-based molecules as novel integrin alpha(v)beta3 antagonists. Part 2: synthesis of potent alpha(v)beta3/alpha(IIb)beta3 dual antagonists. AB - We synthesized 4-aminopiperidine derivatives of our prototype integrin alpha(v)beta3 antagonist 1 in an attempt to increase the activity and water solubility. Introduction of one or two hydrophilic moieties into the central aromatic ring and/or the benzene ring at the C-terminus of 1 increased water solubility and enhanced inhibition of cell adhesion. The results of a structure activity relationships (SAR) study indicated that the torsion angle between the central aromatic ring and the piperidine ring, and the acidity at the sulfonamide moiety, might be important for alpha(v)beta3 receptor binding activity. Some of these compounds are novel and potent alpha(v)beta3/alpha(IIb)beta3 dual antagonists with acceptable water solubility and a satisfactory early absorption, distribution, metabolism, excretion, and toxicity (ADMET) profile. PMID- 16309913 TI - NCA nucleophilic radiofluorination on substituted benzaldehydes for the preparation of [18F]fluorinated aromatic amino acids. AB - Nucleophilic aromatic substitution is a challenging task in radiochemistry. Therefore, a thorough evaluation and optimisation of this step is needed to provide a satisfactory tool for the routine preparation of [(18)F]fluorinated aromatic amino acids. Two methods, already proposed elsewhere, were evaluated and improved. The yields for the radiofluorination were increased whereas activity loss during solid phase extraction was observed. Radiochemical yields for the two methods were 92.7+/-5.5% (method 1) and 92.1+/-12.3% (method 2) for conversion and 11.1+/-2.8% (method 1) and 34.8+/-0.6% (method 2) for purification, respectively. In total, we demonstrate an optimised method for the preparation of this important class of [(18)F]fluorinated synthons for PET. PMID- 16309914 TI - Performance of a whole-body counter with five high-purity germanium detectors. AB - A germanium (Ge) detector system has been installed in a whole-body counting facility at NIRS (Japan). The system employs five detectors that have relative efficiencies of more than 80%. A basic performance of the Ge detectors was tested and compared with that of the former NaI(Tl) detector system. Although the Ge detector system has advantages of improved radionuclide identification capability and MDA values, it also has disadvantages of high operation costs and demanding maintenance such as supply of liquid nitrogen. PMID- 16309915 TI - Synthesis and biodistribution of six novel 99mTc complexes of 2 hydroxybenzaldehyde-amino acid Schiff bases. AB - For the purpose of developing novel diagnostic pharmaceuticals of 99mTc-labeled small-size complexes, six novel complexes of 99mTc-2-hydroxybenzaldehyde-amino acid Schiff bases were designed and synthesized, and their biodistributions in mice were investigated. All the compounds were obtained in radiochemical yields higher than 90% at optimal conditions, and poor uptakes in muscle, brain, heart and tumor were commonly observed with rapid blood clearance. Potentiality was revealed of good kidney imaging by 2-hydroxybenzaldehyde-alanine (L2) complex within 40 min post-injection. Good bone uptake of 2-hydroxybenzaldehyde-histidine (L4) and 2-hydroxybenzaldehyde-aspartic acid (L5) complexes, high spleen accumulation of 2-hydroxybenzaldehyde-glycine (L1) and 2-hydroxybenzaldehyde cysteine (L3) complexes, and non-specific biodistribution of 2 hydroxybenzaldehyde-glutamic acid (L6) complex were demonstrated. PMID- 16309916 TI - Epileptiform activity preferentially arises outside tumor invasion zone in glioma xenotransplants. AB - Seizures occur commonly with brain tumors. The underlying mechanisms are not understood. We analyzed network and cellular excitability changes in tumor invaded and sham neocortical tissue in vitro using a rat glioblastoma model. Rat C6 glioma cells were transplanted into rat neocortex, yielding diffusely invading gliomas resembling human glioblastomas. We hypothesized that network excitability would increase in regions neighboring the tumor, and that initiation of epileptic discharges might be correlated to a higher density of intrinsically bursting neurones. Voltage-sensitive dye imaging revealed epileptic activity to be initiated in paratumoral zones (1-2 mm from main tumor mass), in contrast to control tissue, where epileptic foci appeared randomly throughout the neocortex. Neuronal firing patterns revealed significantly more intrinsically bursting neurones within these initiation zones than in regions directly adjacent to the tumor or in control tissue. We conclude that gliomas are associated with a higher density of intrinsically bursting neurones, and that these may preferentially initiate epileptiform events. PMID- 16309917 TI - Narrow-band UVB induces apoptosis in human keratinocytes. AB - Narrow-band ultraviolet (NB-UVB) phototherapy emits mostly 311/312 nm light and is commonly used in the treatment of inflammatory skin disorders. As a source of UVB irradiation, NB-UVB causes apoptosis in T lymphocytes but its effects on keratinocytes are unknown. Herein, we have investigated the ability of NB-UVB to induce apoptosis in keratinocytes. Two types of human keratinocytes, primary and immortalized, were exposed to NB-UVB and broad-band UVB (BB-UVB; 315-280 nm) and tested for apoptosis. Both UVB light sources induced apoptosis in keratinocytes as determined by the presence of DNA ladders, although NB-UVB required approximately ten fold higher doses; NB-UVB (1000 mJ/cm2) and BB-UVB (125 mJ/cm2). By comparison, lower doses of NB-UVB (750 mJ/cm2) induced apoptosis in T lymphocytes, suggesting cell type specificity for NB-UVB induced apoptosis. Approximately, 50% or more of the cells underwent apoptosis when exposed to NB UVB or BB-UVB as revealed by TUNEL assay. Electron micrographs showed that NB-UVB irradiated keratinocytes contained marked chromatin condensation, extensive cytoplasmic vacuolization and fragmentation of the nuclear envelope. Furthermore, Western blot analysis confirmed the presence of activated products of caspase 3 in keratinocytes that received apoptotic doses of NB-UVB. This study defines conditions by which NB-UVB irradiation causes apoptosis in keratinocytes. PMID- 16309918 TI - Adipose tissue as source and target for novel therapies. AB - The physiology of adipose tissue has a key role in the pathogenesis of the metabolic syndrome and related cardiovascular disorders. Three main functions of adipocytes have been proposed to explain this role: the secretion of adipocyte derived hormones (so-called adipokines), energy dissipation/thermogenesis, and energy storage. There is mounting evidence that this repertoire of actions and reactions contributes to whole-body glucose and energy homeostasis, the control of blood pressure, immune-system function, haemostasis and atherosclerosis. In this article we highlight the most recent examples of adipocyte-based therapies and discuss future pharmacological options for exploiting this triad of adipocyte functions. PMID- 16309919 TI - Aquaporin-1: a novel promoter of tumor angiogenesis. AB - Angiogenesis is critical to solid tumor growth; therefore, understanding how blood vessels form within a developing tumor has become a principal, yet challenging, objective. The recent discovery that deletion of the water channel protein, aquaporin-1, impairs tumor angiogenesis provides novel insights into the mechanisms governing the neovascularization and growth of tumors. PMID- 16309921 TI - Expression of the human activin type I and II receptor extracellular domains in Pichia pastoris. AB - Methods for the expression in Pichia pastoris and purification of the human activin receptor type I and II extracellular domains (ARIa/ARIb-ECDs, ARIIA/ARIIB ECDs) are described. Key experimental aspects are also documented of the vector transformation methodology and the binding characteristics of these ECDs with activin A and inhibin. The cDNA constructs for these ECDs contained a C-terminal His6-tag with either the native signal (N) or the yeast alpha mating factor (alphaMF) sequence and were introduced into the pPICZ expression vector either as a single-copy or as a four-copy expression cassette. Hyper-resistant transformants (zeo(R): 500 microg/mL) generated from the cassette containing a single copy of the expression vector gave the stronger signal intensity with a DNA dot-blot screening assay. These transformants also produced higher quantities of the corresponding recombinant protein compared to transformants using the four copy cassette vector. All receptor-ECD proteins expressed were found to be heterogeneously glycosylated, whereby the ARIIA-ECD and ARIIB-ECD had undergone two Asn-linked glycosylation events and the ARIb-ECD a single event. By SDS-PAGE, the de-glycosylated proteins migrated larger than the expected core size, indicating that they may have undergone O-linked glycosylation. Biacore-based procedures with the glycosylated and de-glycosylated ARIIA-ECD were employed to determine the kinetic and equilibrium binding parameters for the interaction with activin A and inhibin. The glycosylated ARIIA-ECD bound to activin A with a KD of 11.9 nM and inhibin with a KD of 21.1 nM. Although glycosylation of ARIIA-ECD was not strictly required for high affinity interactions with activin A or inhibin, it markedly improved the overall stability of the ARIIA-ECD. PMID- 16309920 TI - Effects of ghrelin and synthetic GH secretagogues on the cardiovascular system. AB - Ghrelin, a newly discovered endogenous hormone that is produced by the stomach, and synthetic peptides have been identified recently as potent growth-hormone secretagogues. This effect is exerted through interaction with a specific G protein-coupled receptor, GHS-R1a, which is expressed mainly in the hypothalamus pituitary complex. A study of the peripheral distribution of GHS receptors has shown that it is also present in cardiovascular tissue, which has led to the exploration of the cardiovascular functions of ghrelin and synthetic, growth hormone-releasing peptides. These ligands have several cardiovascular activities, including a cardioprotective effect against myocardial ischemia, and vasoactive and cardiotropic effects in both experimental models and humans. These effects are mediated by the interaction of these ligands with binding sites, including GHS-1Ra, for which the signalling pathways are not documented fully. Identification of the cardiac and vascular binding sites for ghrelin and synthetic, growth-hormone-releasing peptides will provide new perspectives for treating cardiovascular diseases with these ligands. PMID- 16309922 TI - In vivo motion transmission in the inactive gastrocnemius medialis muscle-tendon unit during ankle and knee joint rotation. AB - The purposes of this study were: (a) to quantify the influence of passive ankle and knee joint angular displacement on the estimated mechanical and architectural properties of the gastrocnemius medialis (GM) muscle-tendon unit, and (b) to determine the strain distribution of separate structures (tendon, aponeurosis and fascicle) during passive lengthening of the GM muscle-tendon unit at rest. Ten male subjects participated in the study. The passive ankle and knee joint movements were performed on an isokinetic dynamometer. The kinematics of the left leg were recorded using the Vicon 624 system with 8 cameras. Two ultrasound probes were used to examine the elongation of the tendon, the aponeurosis, the fascicles and the angle of pennation of the GM. To calculate the elongation of the GM muscle-tendon unit the Achilles tendon path was reconstructed using a series of small reflective markers. The results show that the passive ankle joint angular displacement has a considerable influence on the elongation of the tendinous and architectural structures of the GM muscle-tendon unit. In contrast, the influence of knee joint angular displacement on the GM fascicle length and pennation angle becomes relevant only at knee angles greater than 144 degrees . The contribution of the tendon to the elongation of the GM muscle-tendon unit at rest is relevant because of its greater resting length in comparison to the resting length of the GM fascicles. The results indicate the existence of slackness in the inactive GM muscle-tendon unit between 121 degrees and 107 degrees ankle angle and between 65 degrees and 144 degrees knee angle. PMID- 16309923 TI - Anomalous functional organization of barrel cortex in GAP-43 deficient mice. AB - Growth associated protein 43 (GAP-43), found only in the nervous system, regulates the response of neurons to axon guidance signals. It is also critical for establishing normal somatotopy. Mice lacking GAP-43 (KO) show aberrant pathfinding by thalamocortical afferents, and do not form cortical whisker/barrels. GAP-43 heterozygous (HZ) mice show more subtle deficits--delayed barrel segregation and enlarged barrels at postnatal day 7. Here, we used cortical intrinsic signal imaging to characterize adult somatotopy in wildtype (WT), GAP-43 KO, and HZ mice. We found clear foci of activation in GAP-43 KO cortex in response to single-whisker stimulation. However, the KO spatial activation patterns showed severe anomalies, indicating a loss of functional somatotopy. In some cases, multiple foci were activated by single whiskers, while in other cases, the same cortical zone was activated by several whiskers. The results are consistent with our previous findings of aberrant pathfinding and clustering by thalamocortical afferent axons, and absence of barrel patterning. Our findings indicate that cortex acts to cluster afferents from a given whisker, even in the absence of normal topography. By contrast, single-whisker stimulation revealed normal adult topographic organization in WT and HZ mice. However, we found that functional representations of adult HZ barrels are larger than those found in WT mice. Since histological HZ barrels recover normal dimensions by postnatal day 26, the altered circuit function in GAP-43 HZ cortex could be a secondary consequence of the rescue of barrel dimensions. PMID- 16309924 TI - Discrepancies in phylogeographical patterns of two European anglerfishes (Lophius budegassa and Lophius piscatorius). AB - In order to investigate the relative importance of historical processes and life history traits in shaping the present-day genetic structure of European anglerfishes, 382 Lophius piscatorius and 134 Lophius budegassa were sequenced on the 5' end of the mitochondrial control region. Both species showed a limited genetic structure and some evidence of a demographic expansion that probably occurred not at the end of the Last Glacial Maximum, but earlier in the Pleistocene. The main discrepancy between the two anglerfishes concerned the genetic structure between Atlantic and Mediterranean populations, with weak but significant differentiation observed only in L. budegassa. This genetic structure was congruent with the existence of a phylogeographical break previously reported in several marine species across the Almeria-Oran front. The contrast observed between both anglerfishes was supposed to be induced by a possible more ancient (re)colonisation of the Mediterranean Sea by L. budegassa. Finally, the limited genetic structure and lack of isolation by distance observed in both species suggested high larval dispersal capacities that probably overwhelm the influence of the bathymetric distribution range on migrations of adults and juveniles. PMID- 16309925 TI - Perinatal seizures preferentially protect CA1 neurons from seizure-induced damage in prepubescent rats. AB - Neonatal seizures may increase neuronal vulnerability later in life. Therefore, status epilepticus was induced with kainate (KA) during the first and second postnatal (P) weeks to determine whether early seizures shift the window of neuronal vulnerability to a younger age. KA was injected (i.p.) once (1x KA) on P13, P20 or P30 or three times (3 x KA), once on P6 and P9, and then either on P13, P20 or P30. After 1x KA, onset to behavioral seizures increased with age. Electroencephalography (EEG) showed interictal events appeared with maturation. After 3 x KA, spike number, frequency, spike amplitude, and high-frequency synchronous events and duration were increased at P13 when compared to age matched controls. In contrast, P20 and P30 rats had decreases in EEG parameters relative to P20 and P30 rats with 1x KA despite that these animals had the same history of perinatal seizures on P6 and P9. In P13 rats with 1x KA, silver impregnation, hematoxylin/eosin and TUNEL methods showed no significant hippocampal injury and damage was minimal with 3 x KA. In contrast, P20 and P30 rats with 1x KA had robust eosinophilic or TUNEL positive labeling and preferential accumulation of silver ions within inner layer CA1 neurons. After 3 x KA, the CA1 but not CA3 of P20 and P30 rats was preferentially protected following 3 or 6 days. Although paradoxical changes occur in the EEG with maturation, the results indicate that early perinatal seizures do not significantly shift the window of hippocampal vulnerability to an earlier age but induce a tolerance that leads to long-term neuroprotection that differentially affects endogenous properties of CA1 versus CA3 neurons. PMID- 16309926 TI - Is primidone the drug of choice for epileptic patients with QT-prolongation? A comprehensive analysis of literature. AB - Sudden unexplained/unexpected death (SUDEP) in epilepsy is a major cause of death accounting for 7-17% of the mortality among epileptic patients. Prolongation of QT-interval has been issued as a major mechanism in SUDEP since it is associated with fatal cardiac arrhythmias. This condition may be further precipitated by anti-epileptic treatment. Despite thorough literature research, we did not find any reports suggesting that primidone is responsible for QT-prolongation. On the contrary, all the retrieved reports addressed that the drug shortened QT-interval and corrected signs and symptoms of the underlying disease. PMID- 16309927 TI - Raised chondroitin sulfate epitopes and hyaluronan in serum from rheumatoid arthritis and osteoarthritis patients. AB - OBJECTIVES: Serum hyaluronan (HA) and chondroitin sulfate (CS) epitopes WF6 and 3B3 (+) were determined to investigate disease association in patients with osteoarthritis (OA), rheumatoid arthritis (RA) and healthy controls. METHODS: Specific assays for HA and CS epitopes WF6 and 3B3 (+) were established and applied to a cross-sectional study of serum samples from patients (96 OA, 57 RA and 50 healthy controls). RESULTS: Both CS epitopes were increased in serum of many OA and RA patients and average levels were significantly above in healthy controls. In contrast serum HA was increased in RA, but only in few OA patients. CONCLUSIONS: CS epitopes WF6 and 3B3 (+) are raised in serum of patients with both OA and RA and were thus distinct from serum HA. The results suggest that OA may be detected systemically as well as RA. The range of levels of CS epitopes detected in OA and RA was wide and correlation with any aspect of disease activity is yet to be determined. PMID- 16309928 TI - Efficacy of methylsulfonylmethane (MSM) in osteoarthritis pain of the knee: a pilot clinical trial. AB - OBJECTIVE: Osteoarthritis (OA) is the most common form of arthritis and the second most common cause of long-term disability among middle-aged and older adults in the United States. Methylsulfonylmethane (MSM) is a popular dietary supplement used as a single agent and in combination with other nutrients, and purported to be beneficial for arthritis. However, there is paucity of evidence to support the use of MSM. METHODS: A randomized, double-blind, placebo controlled trial was conducted. Fifty men and women, 40-76 years of age with knee OA pain were enrolled in an outpatient medical center. Intervention was MSM 3g or placebo twice a day for 12 weeks (6g/day total). Outcomes included the Western Ontario and McMaster University Osteoarthritis Index visual analogue scale (WOMAC), patient and physician global assessments (disease status, response to therapy), and SF-36 (overall health-related quality of life). RESULTS: Compared to placebo, MSM produced significant decreases in WOMAC pain and physical function impairment (P<0.05). No notable changes were found in WOMAC stiffness and aggregated total symptoms scores. MSM also produced improvement in performing activities of daily living when compared to placebo on the SF-36 evaluation (P<0.05). CONCLUSION: MSM (3g twice a day) improved symptoms of pain and physical function during the short intervention without major adverse events. The benefits and safety of MSM in managing OA and long-term use cannot be confirmed from this pilot trial, but its potential clinical application is examined. Underlying mechanisms of action and need for further investigation of MSM are discussed. PMID- 16309929 TI - The effect of an educational program to improve health-related quality of life in patients with osteoarthritis on waiting list for total knee replacement: a randomized study. AB - OBJECTIVE: To evaluate the effect of therapeutic education and functional readaptation (TEFR) on health-related quality of life (HRQL) in patients diagnosed with osteoarthritis on a waiting list for total knee replacement (TKR). METHODS: Randomized controlled trial of 9 months duration was conducted. One hundred consecutive outpatients (71 females, mean age 71 years (range 50-86), mean disease duration 11.84+/-10.52 months) were included. Patients were randomized in two groups. The intervention group received TEFR added to conventional (pharmacological) treatment (n=51). The control group received conventional (pharmacological) treatment only (n=49). The main outcome variable was self-reported HRQL measured by the Spanish version of Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Secondary outcomes were general HRQL measured by Short Form Health Survey general questionnaire (SF-36), number of visits to general physicians and their cost. Assessments were done at baseline and at 9 months. MAIN RESULTS: Eighty patients completed the study. Significant improvement in the WOMAC function was found at 9 months in the TERF group with respect to the control group (P=0.035). Consumption of analgesics increased significantly in the TERF group compared with controls (P=0.036). Significant improvements in pain (P=0.027) measured by WOMAC and in bodily pain (P=0.043) and physical function (P=0.031), measured by SF-36, were observed in the intervention group with respect to baseline. CONCLUSIONS: The function dimension measured by WOMAC of patients who received both pharmacological treatment and TERF improved with respect to the control group receiving only pharmacological treatment. This suggests that a program of TEFR during the period on the waiting list for TKR may reduce the negative impact of this situation. PMID- 16309931 TI - 5-Azacytidine decreases fragmentation of nuclear DNA and pigment formation in first leaf cells of barley seedlings. AB - Realization of programmed cell death in senescence represents an activation/inactivation of the respective gene. Enzymatic methylation of nuclear DNA with the creation of 5methylcytosine is one of the mechanisms, which can regulate gene activity in animal and plant cells. 5Azacytidine (5azaC) acts as an inhibitor of DNA methylation, and induces expression of a range of some genes including genes responsible for senescence. Fragmentation of nuclear DNA is one of the hallmarks of programmed cell death in apoptosis pathway in plant cells. The influence of 5azaC (100 microg/ml) on nuclear DNS amount and its fragmentation in the first leaf cells of barley was studied. It was shown that in the first leaf cells of barley seedlings there is an apoptosis pathway of programmed cell death. It was also observed that nuclear DNA fragmentation under the 5azaC influence is strongly inhibited, and the DNA amount in the first leaf increases. Synthesis and destruction of chlorophyll also play a significant role in programmed cell death in plants. It was shown that under the 5azaC influence, the absorption spectrum of a pigment does not change in leaves and coleoptiles in the light, whereas in the dark condition, these pigments are not created under the 5azaC influence. PMID- 16309932 TI - Hunting the mechanisms of self-renewal of immortal cell populations by means of real-time imaging of living cells. AB - The causes of the indefinite propagation of immortalized cell populations remain insufficiently understood, that hinders the research of such fundamental processes as ageing and cancer. In this study the interrelations between clonal proliferation and abnormalities of mitotic divisions in the immortalized cell line established from the mouse embryo were investigated with the aid of computerized microscopy of living cells. 3 mitoses with three daughter cells and 7 asymmetric mitoses which generated two daughter cells of conspicuously different sizes were registered among 71 mitotic divisions in the individual cell genealogy. Abnormal mitotic divisions either did not slow the proliferation in cell clones compared with progenies of cells that divided by means of normal mitoses or were followed by the acceleration of divisions in consecutive cell generations. These data suggest that abnormal mitotic divisions may contribute to the maintenance of the immortalized state of cell populations by means of generating chromosomal instability. PMID- 16309933 TI - Nitrative stress through formation of 8-nitroguanosine: insights into microbial pathogenesis. AB - Reactive oxygen and nitrogen species, respectively, mediate oxidative and nitrative stresses by means of oxidation and nitration of various biomolecules including proteins, lipids, and nucleic acids. We have observed nitric oxide (NO) dependent formation of 8-nitroguanosine and 3-nitrotyrosine during microbial infection, and we determined that both 8-nitroguanosine and 3-nitrotyrosine are useful biomarkers of nitrative stress. Of importance, however, is the great difference in biological characteristics of these two nitrated compounds. 8 Nitroguanosine has unique biochemical and pharmacological properties such as redox activity and mutagenic potential, which 3-nitrotyrosine does not. In this review, we discuss the mechanism of nitrative stress occurring during microbial infections, with special emphasis on biological functions of 8-nitroguanosine formed via NO during the host response to pathogens. These findings provide insights into NO-mediated pathogenesis not only of viral infections but also of many other diseases. PMID- 16309934 TI - Nitric oxide-mediated inhibition of DNA synthesis was attenuated in hypertrophied neonatal rat ventricular myocytes. AB - The antiproliferative action of nitric oxide (NO) has been well established and increased production was reported in the infarcted rat heart. Concomitantly, increased DNA synthesis and hyperplasia of cardiac myocytes were documented in the hypertrophied myocardium. Despite these observations, the effect of NO on DNA synthesis in hypertrophied cardiac myocytes remains unexamined. Hypertrophy of the non-infarcted left ventricle (NILV) in 1-week post-MI rats was characterized by the increased prepro-ANP and reduction of alpha-myosin heavy chain protein expression. Inducible NO synthase was expressed in the NILV and associated with a concomitant attenuation of MnSuperoxide dismutase protein content. The latter data suggest that an antiproliferative action of NO in the hypertrophied NILV may proceed via either a cyclic GMP-dependent pathway and/or facilitated by a peroxynitrite-dependent mechanism. In neonatal rat ventricular myocytes (NNVM), the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) promoted a dose-dependent attenuation of DNA synthesis via a cyclic GMP-independent pathway. The permeable superoxide dismutase mimetic and peroxynitrite scavenger MnTBAP abrogated SNAP dependent attenuation of DNA synthesis in NNVM. MnTBAP failed to inhibit SNAP mediated recruitment of extracellular signal regulated kinase 1/2 (ERK1/2) but partially attenuated p38 phosphorylation. In hypertrophied NNVM induced by norepinephrine, SNAP-mediated peroxynitrite-dependent inhibition of DNA synthesis, ERK1/2 and p38 phosphorylation were significantly attenuated. Collectively, these data suggest that despite a favourable environment for NO and subsequent peroxynitrite generation in the NILV, hypertrophied cardiac myocytes may be partially refractory to their biological actions. PMID- 16309935 TI - Aggravating factors in the development of ocular abnormalities in farmed Atlantic halibut (Hippoglossus hippoglossus). AB - Cystic lesions in the eyes of farmed halibut with a number of ocular sequelae have been reported previously with evidence that the condition can be associated with increased activity of choroidal carbonic anhydrase and elevated oxygen tension in the aqueous humour of affected fish. These changes may be compared to the 'bends' in human divers which are characterized by elevated circulating levels of nitrogen together with aggravating factors such as physical exercise. It is postulated that bubble formation and subsequent development of cysts in halibut require not only high aqueous humour oxygen tension but also the imposition of factors such as handling or intraspecific aggression which lead to bubble formation. A significantly higher incidence of lesions was noted in fish handled, weighed and measured every two weeks compared with a control group of unhandled fish, suggesting that physical activity associated with handled and possibly also intraspecific aggression is a factor in generation of ocular lesions in these fish. PMID- 16309936 TI - Symmetry-based recoupling of 17O-1H spin pairs in magic-angle spinning NMR. AB - We have performed magic-angle-spinning solid-state NMR experiments in which protons are recoupled to oxygen-17 nuclei by applying a symmetry-based recoupling sequence at the proton Larmor frequency. Two-dimensional quadrupole-dipole correlation spectra are produced, in which the second-order quadrupolar shift of the oxygen-17 central transition is correlated with the recoupled heteronuclear dipole-dipole interaction. These spectra are sensitive to the relative orientation of the electric field gradient at the site of the oxygen-17 nucleus and the O-H internuclear vector. We also demonstrate experiments in which polarization is transferred from protons to oxygen-17, and show that oxygen-17 signals may be selected according to the protonation state of the oxygen site. We discuss the small observed value of the heteronuclear dipolar splitting in the central-transition oxygen-17 spectra. PMID- 16309937 TI - Effect of the acute crowding stress on the rat brown adipose tissue metabolic function. AB - Our previous results have shown that metabolic and thermal stressors influence interscapular brown adipose tissue (IBAT) metabolic activity by increasing oxygen consumption and, consequently, altering the toxic reactive oxygen species (ROS) production and the antioxidative system activity. Since there is not enough evidence about the effect of psychosocial stressors on these processes, we studied the effect of acute crowding stress on the IBAT and hypothalamic monoamine oxidase (MAO) activity as well as IBAT antioxidative enzymes, manganese (MnSOD), copper-zinc superoxide dismutase (CuZnSOD) and catalase (CAT), as the relevant indicators of IBAT metabolic alternations under the stress exposure and the returning of animals to control conditions. The results indicated that acute crowding stress did not change the hypothalamic and IBAT MAO activities, the generation of ROS and, consequently, the IBAT CuZnSOD and CAT activities. However, all three antioxidative enzymes were affected only after the recovery period. It seems that peripheral overheating of rats during acute crowding changes the stress nature, by becoming more thermal than psychosocial and by suppression the hypothalamic efferent pathways involved in the IBAT thermogenesis regulation. However, it seems that returning of the animals to the control conditions after the stress termination causes the reactivation of IBAT thermogenesis with tendency to normalise the body temperature. PMID- 16309938 TI - Seasonal patterns in body temperature of free-living rock hyrax (Procavia capensis). AB - Rock hyrax (Procavia capensis) are faced with large daily fluctuations in ambient temperature during summer and winter. In this study, peritoneal body temperature of free-living rock hyrax was investigated. During winter, when low ambient temperatures and food supply prevail, rock hyrax maintained a lower core body temperature relative to summer. In winter body temperatures during the day were more variable than at night. This daytime variability is likely a result of body temperatures being raised from basking in the sun. Body temperatures recorded during winter never fell to low levels recorded in previous laboratory studies. During summer ambient temperatures exceeded the thermoneutral zone of the rock hyrax throughout most of the day, while crevice temperatures remained within the thermoneutral zone of rock hyrax. However, in summer variation in core body temperature was small. Minimum and maximum body temperatures did not coincide with minimum and maximum ambient temperatures. Constant body temperatures were also recorded when ambient temperatures reached lethal limits. During summer it is likely that rock hyrax select cooler refugia to escape lethal temperatures and to prevent excessive water loss. Body temperature of rock hyrax recorded in this study reflects the adaptability of this animal to the wide range of ambient temperatures experienced in its natural environment. PMID- 16309939 TI - [Brachytherapy for soft tissue sarcomas. Technique and therapeutic indications]. AB - The role of brachytherapy in the therapeutic strategy of soft tissue sarcomas has been difficult to assess due to the fact it has been during a long time institution-specific. However, the experience gained specially with low dose-rate treatments has allowed formulating guidelines for technical rules and patient selection. Brachytherapy used as an isolated treatment combined with surgery has proven to be effective in high-grade sarcomas, compared to surgery alone, in a unique randomized trial. Beside, a number of data have shown that the combination of brachytherapy and external beam irradiation could be superior to brachytherapy or external beam irradiation alone in the treatment of tumors with high-grade, or central location, locally advanced or on contact with neurovascular structures. Brachytherapy alone, combined with surgery, is of interest for conservative treatment of recurrent sarcomas occurring in previously irradiated areas. The occurrence of side effects remains acceptable if technical guidelines are followed, for surgery as well as for brachytherapy, using the most modern available treatment techniques. Brachytherapy should be at the moment an integrated part of the multidisciplinary treatment of soft tissue sarcomas. PMID- 16309940 TI - [Renal medullary carcinoma: a case report]. AB - Renal medullary carcinoma is an aggressive malignant tumour, recently reported in the literature. It is usually reported in the relatively young patients with drepanocytic trai. Histologically, the tumour is constituted by a tumoral proliferation with diffuse or glandular architecture and inflammatory stroma. The carcinomatous cells have plasmocytoid or rhabdoid aspect. We report a case of 40 years old man who presented macroscopic hematuria. Through this observation and the review of the literature we discuss the anatomoclinical and the prognostic aspects of this exceptional tumour. PMID- 16309941 TI - [Surgery of limb and trunk wall soft tissue sarcoma]. AB - For this review of surgery in soft tissue sarcoma, we reviewed literature and added our personal experience. In front of a soft tissue tumour, the major objective is to think it could be a sarcoma. Diagnosis actually is made by core needle biopsies, but sometimes a surgical biopsy is needed. Surgical resection is better defined since ten years and distinguishes resection R0 (in sano), R1 (microscopic residual disease) and R2 (macroscopic residual disease). Quality of resection is determined collegially by confrontation of surgical and pathological reports according recommendations of the French Sarcoma Group (FSG). The risk of local recurrence depends on resection-type and tumour grade. With the help of the surgical classification from FSG, local prognosis is better understood and treatment can be adapted to individual tumours. In experienced hands, local recurrence can be pushed down to near 10 against 20% formerly. Severe complications after sarcoma surgery occur in 14% of cases. Functional outcome studies have benefited from new scoring systems, which complete each other. Isolated limb perfusion is a method that could save limbs in some desperate situations. CONCLUSION: Due to rarity and treatment specifications, soft tissue sarcoma should be treated in specialised centres. PMID- 16309942 TI - [Hodgkin disease of the nasopharynx: report of three cases]. AB - We report three cases of Hodgkin's disease (HD) involving the nasopharynx. Their clinical presentations, morphological and immunohistochimical features and their therapy modalities are discussed. The patients were aged 36, 41 and 77, presenting with increasing bilateral nasal obstruction in one case and a cervical mass in the two others. Histological study showed mixed cellularity type of HD in all cases. The Reed Sternberg cells expressed both of CD15 and CD30 in one case, and only one of them in the other cases. In one case, LMP1 was detected, CD20 and CD3 were not. HD of nasopharynx should be differentiated from EBV-associated lymphoproliferations. The treatment is based on radiotherapy that can be associated to neoadjuvant chemotherapy if nodes are involved. PMID- 16309943 TI - Genetic basis of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a multifactorial disease due to a combination of genetic and environmental factors. Identification of the genetic factors involved in the pathogenesis of RA should open up avenues for developing radical treatment strategies directed at the cause of the disease. The Association de Recherche sur la Polyarthrite (ARP) supports research in this field, in which our group has been involved since 1993. Thanks to this support, considerable progress has been made. Several combinations of susceptibility alleles of various genes are probably involved in the development of RA. Although HLA-DRB1 is the main RA gene, it accounts for only part of the familial risk for RA. HLA-DRB1 alleles are neither necessary nor sufficient to cause the development of RA in a given individual. Several genome scans conducted in populations from France, Japan, North America and UK have confirmed the role of the HLA region and suggested several other susceptibility loci. Association studies support a role for several genes, including TNFR2, PADI4, SLC22A4, RUNX1, and PTPN22. However, the imperfect matching of cases and controls requires that confirmation of these results be obtained. To confirm that a gene confers susceptibility to RA, the association must be replicated in several independent studies and, more importantly, evidence of genetic linkage must be obtained in family studies. The identification of genetic factors conferring susceptibility to RA will open up new avenues toward radical treatments for RA and may help to optimize the diagnostic, prognostic, and pharmacogenetic management of today's patients with RA. PMID- 16309944 TI - Moving single bubble sonoluminescence in phosphoric acid and sulphuric acid solutions. AB - The phenomenon of sonoluminescence still presents some unsolved aspects. Recently [Y.T. Didenko, K. Suslick, Molecular Emission during Single Bubble Sonoluminescence, Nature 407 (2000) 877-879.], it was found that a single cavitating air bubble in polar aprotic liquids (including formamide and adiponitrile) can produce very strong sonoluminescence while undergoing macroscopic translation movements in the resonator, a condition known as moving single bubble sonoluminescing (MSBSL). Here we describe some experiments conducted in aqueous solutions of phosphoric and sulphuric acid. In these liquid media, it is possible to reproduce MSBSL and luminescence is emitted even if a trapped bubble is subjected to a strong shape instability, named in the literature "jittering phase". When a moving and luminescing bubble was present and the acoustic pressure gradually increased, we observed the generation of a discrete lattice of trapped bubbles. The bubbles in the lattice emit very intense light flashes and can change their position while maintaining the overall spatial distribution in time. Some preliminary results, obtained from Mie-scattering and measurements of relative light intensity, are reported. PMID- 16309945 TI - Actin is not an essential component in the mechanism of calcium-triggered vesicle fusion. AB - Actin has been suggested as an essential component in the membrane fusion stage of exocytosis. In some model systems disruption of the actin filament network associated with exocytotic membranes results in a decrease in secretion. Here we analyze the fast Ca2+-triggered membrane fusion steps of regulated exocytosis using a stage-specific preparation of native secretory vesicles (SV) to directly test whether actin plays an essential role in this mechanism. Although present on secretory vesicles, selective pharmacological inhibition of actin did not affect the Ca2+-sensitivity, extent, or kinetics of membrane fusion, nor did the addition of exogenous actin or an anti-actin antibody. There was also no discernable affect on inter-vesicle contact (docking). Overall, the results do not support a direct role for actin in the fast, Ca2+-triggered steps of regulated membrane fusion. It would appear that actin acts elsewhere within the exocytotic cycle. PMID- 16309946 TI - Regulators of angiogenesis and strategies for their therapeutic manipulation. AB - Angiogenesis provides a mechanism by which delivery of oxygen and nutrients is adapted to compliment changes in tissue mass or metabolic activity. However, maladaptive angiogenesis is integral to the process of several diseases common in Western countries, including tumor growth, vascular insufficiency, diabetic retinopathy and rheumatoid arthritis. Understanding the process of capillary growth, including the identification and functional analyses of key pro- and anti angiogenic factors, provides knowledge that can be applied to improve/reverse these pathological states. Initially, angiogenesis research focused predominantly on vascular endothelial growth factor (VEGF) as a main player in the angiogenesis cascade. It is apparent now that participation of multiple angiogenic factors and signal pathways is critical to enable effective growth and maturation of nascent capillaries. The purpose of this review is to focus on recent progress in identifying angiogenesis signaling pathways that show promise as targets for successful induction or inhibition of capillary growth. The strategies applied to achieve these contradictory tasks are discussed within the framework of our existing fundamental knowledge of angiogenesis signaling cascades, with an emphasis on comparing the employment of distinctive tactics in modulation of these pathways. Innovative developments that are presented include: (1) inducing a pleiotropic response via activation or inhibition of angiogenic transcription factors; (2) modulation of nitric oxide tissue concentration; (3) manipulating the kallikrein-kinin system; (4) use of endothelial progenitor cells as a means to either directly contribute to capillary growth or to be used as a vehicle to deliver "suicide genes" to tumor tissue. PMID- 16309947 TI - A metric for the cognitive map: found at last? AB - A network of brain areas collectively represent location, but the underlying nature of this "cognitive map" has remained elusive. A recent study reports that the activity patterns of some entorhinal cortical neurons form a remarkably regular array of evenly spaced peaks across the surface of the environment. These "grid cells" might be the basis of a metric used for calculating position, and their discovery could greatly advance our understanding of how navigational computations are performed. PMID- 16309948 TI - RGD-based strategies for selective delivery of therapeutics and imaging agents to the tumour vasculature. AB - During the past decade, RGD-peptides have become a popular tool for the targeting of drugs and imaging agents to alphavbeta3-integrin expressing tumour vasculature. RGD-peptides have been introduced by recombinant means into therapeutic proteins and viruses. Chemical means have been applied to couple RGD peptides and RGD-mimetics to liposomes, polymers, peptides, small molecule drugs and radiotracers. Some of these products show impressive results in preclinical animal models and a RGD targeted radiotracer has already successfully been tested in humans for the visualization of alphavbeta3-integrin, which demonstrates the feasibility of this approach. This review will summarize the structural requirements for RGD-peptides and RGD-mimetics as ligands for alphavbeta3. We will show how they have been introduced in the various types of constructs by chemical and recombinant techniques. The importance of multivalent RGD-constructs for high affinity binding and internalization will be highlighted. Furthermore the in vitro and in vivo efficacy of RGD-targeted therapeutics and diagnostics reported in recent years will be reviewed. PMID- 16309949 TI - Evaluation of genotoxic effects of Apitol (cymiazole hydrochloride) in vitro by measurement of sister chromatid exchange. AB - Apitol, with cymiazole hydrochloride as the active ingredient, is used in bee keeping against the ectoparasitic mite Varroa destructor. The preparation was evaluated for genotoxicity in cultured human peripheral blood lymphocytes. Sister chromatid exchange, the mitotic index and the cell proliferation index were determined for three experimental concentrations of Apitol (0.001, 0.01 and 0.1 mg/ml). All concentrations significantly (p < 0.001) increased the mitotic index (MI = 7.35+/-0.18%, 8.31+/-0.20% and 12.33+/-0.25%, respectively), the proliferative index (PI = 1.83+/-0.01, 1.84+/-0.01 and 1.88+/-0.02, respectively) and the frequency of sister chromatid exchange (SCE = 8.19+/-1.81, 8.78+/-1.80 and 13.46+/-1.88, respectively), suggesting that cymiazole hydrochloride has genotoxic potential. PMID- 16309950 TI - A procedure for selecting and culturing subpopulations of neurons from rat dorsal root ganglia using magnetic beads. AB - Current protocols for preparing primary sensory neuron cultures are inadequate when studying individual subpopulations of dorsal root ganglion (DRG) neurons. The DRG is made up of a heterogeneous population of cells, making it difficult to study treatment effects on any given population in mass cultures. Thus, we describe a procedure using magnetic beads from Dynal to select and plate viable populations of neurons based on expression of specific cell surface markers. We show that, by the use of the lectin IB4, we can select a highly enriched viable subpopulation of GDNF-responsive DRG neurons, leaving a viable population of non selected IB4-ve, Trk+ve neurons. Key factors for successful cultures are (i) quick and careful dissection of DRGs from 4- to 5-week-old Sprague-Dawley rats, (ii) adequate removal of debris and non-neuronal contamination and (iii) gentle handling of bead-bound cells during selection. PMID- 16309951 TI - Piloting a web-based continuing professional development program for asthma education. AB - PURPOSE: Continuing professional development is an integral component of modern medical practice, yet traditional educational methods are impractical for many Primary Care Physicians. Web-based programs may fulfill the requirements of busy practitioners who have difficulty attending formal education sessions. METHODS: We piloted the use of a learning management system to deliver asthma education materials to Primary Care Physicians in both Australia and Italy in their native languages. Each group of Physicians accessed an education module which contained content pages, self-tests, a quiz and a survey. Details of how the Physicians used the system, their preferences and performance on the assessment were monitored. RESULTS: The learning management system was well received by both Italian and Australian Physicians. Thirty-eight (18 Australian, 20 Italian) Physicians used the system. Participants visited an average of 8.8 pages, with a mean time per hit of 2.9 min. Formative assessment was undertaken by 63.2% and summative assessment by 68.4% of participants. There were no substantial differences in performance between Physicians from both countries. Italian physicians tended to use the system after hours whereas Australian Physicians appear to do so between patient visits. CONCLUSIONS: Simple web-based systems are suitable for delivering educational materials to Primary Care Physicians in a manner likely to be used. PMID- 16309952 TI - Influenza virus strains in Nairobi, Kenya. PMID- 16309953 TI - Decreased survival in diabetic patients with heart failure due to systolic dysfunction. AB - BACKGROUND: Prognosis of patients with heart failure (HF) has improved in recent years due to advances in therapy. Whether this is also true for diabetic subjects with HF in clinical practice has not been studied in a prospective manner. METHODS: All patients with HF and left ventricular systolic dysfunction attending the outpatient clinic at our Veteran's Hospital between October 1999 and November 2000 were enrolled in our study and followed prospectively. Electronic medical records were accessed for data on comorbid conditions, medications, echocardiogram results and mortality information. Mean follow-up was 2.7 years. RESULTS: Of 495 patients with HF due to systolic dysfunction enrolled in the study, 293 (59%) had diabetes. Prevalence of hypertension, diuretic use and angiotensin converting enzyme inhibitor use was higher among diabetics. Beta blocker usage was equal and high in both groups (60%). On follow-up, 109/273 (37%) patients in the diabetic group died, compared with 49/202 (24%) in the non diabetic group. Independent predictors of death were diabetes (p<0.005, OR=1.73), age at enrollment (p<0.0001, OR=1.06), serum creatinine (p<0.01, OR=1.44) and diuretic use (p=0.038, OR=1.85). Beta-blocker use was associated with a decreased risk of death on univariate analysis only. CONCLUSIONS: Our results show that diabetic patients with HF continue to have higher mortality than non-diabetic patients with HF despite advances in therapy. PMID- 16309954 TI - Influence of ACE-inhibition and mechanical unloading on the regulation of extracellular matrix proteins in the myocardium of heart transplantation candidates bridged by ventricular assist devices. AB - BACKGROUND: Whether adverse structural changes in the myocardium due to remodelling can be reversed by ventricular assist device (VAD) support in patients with end-stage heart failure is controversial. AIMS: To investigate the effect of VAD support on the extra-cellular matrix. METHODS: We analysed the collagen content in terminal failing ventricles of VAD-patients and donor hearts using 4-hydroxyproline for total collagen and real time RT-PCR for fibronectin (FN), collagen I alpha 1 (Col1A1), III alpha 1 (Col3A1) and TGF beta 1 analysis. RESULTS: Compared to donor hearts we found similar increases in Col1A1 and TGF beta1 but not Col3A1 and FN mRNAs, which were similar in the myocardium from patients receiving a VAD or heart transplant. However, patients receiving ACE-I during VAD-support had lower Col1A1 mRNA content at transplantation. The total collagen content was not influenced by mechanical unloading or by ACE-I medication. CONCLUSION: Mechanical unloading by VAD does not reduce the collagen content of the terminal failing ventricle possibly due to increased TGF beta1 levels. However, Col1A1 production may be reduced by ACE-I medication during VAD support. PMID- 16309955 TI - Relationship between cytokines and tumour markers in patients with chronic heart failure. AB - AIMS: Serum levels of some cytokines and tumour markers are elevated in patients with chronic heart failure (HF). We aimed to investigate the relationship between circulating levels of cytokines and tumour markers in patients with HF. METHODS: We included 35 HF patients and 33 normal controls. HF patients were divided into two groups: mild HF (NYHA class I/II) (n=10) and severe HF (NYHA class III/IV) (n=25). Serum cytokine levels (TNF-alpha, IL-1 beta, IL-6, and IL-10) were measured by ELISA and tumour markers (CA 125, CA 19-9, CA 15-3, CEA and AFP) by chemiluminescent enzyme immunoassay. RESULTS: Serum levels of TNF-alpha, IL-6, and IL-10 as cytokines, and CA 125 and CA 19-9 as tumour markers were significantly higher in HF patients than in normal controls (p<0.0001 for all). Serum levels of TNF-alpha, IL-6 and IL-10 and CA 125 in the severe HF patients were significantly higher than in the mild HF patients (p<0.001 for all). Correlation analysis showed that CA 125 was positively related to TNF-alpha (r=0.624, p<0.001), IL-6 (r=0.671, p<0.001), and IL-10 (r=0.545, p<0.001) in HF. CONCLUSION: These findings show that CA 125 is markedly elevated in patients with HF, and correlates with serum TNF-alpha, IL-6 and IL-10 levels. Therefore, we speculate that among the tumour markers studied, only CA 125 is closely related to the cytokine system. PMID- 16309956 TI - Influence of atrial fibrillation on cardiac brain natriuretic peptide release during haemodynamic stress in heart failure. AB - BACKGROUND: The determinants of release of brain natriuretic peptide (BNP) in heart failure (HF) are incompletely understood, particularly, the effect of heart rhythm and haemodynamic stress. AIMS: To investigate the effect of haemodynamic stress on cardiac BNP release in HF and differentiate this response for atrial fibrillation (AF) and sinus rhythm (SR). METHODS: In 18 HF patients (ejection fraction<40%, 9 in AF and 9 in SR) haemodynamics and BNP levels were measured from arterial and coronary sinus samples at baseline, after 10 min of 20 degrees passive head up tilt (HUT) and after 10 min of isometric handgrip (IHG) exercise. From these data, we calculated a transcardiac BNP gradient and compared results between the AF and SR cohort. RESULTS: During haemodynamic stress in both groups, there were no significance differences in left sided filling pressures. At baseline, there were no differences in BNP measurements between the SR and AF group. The transcardiac BNP gradient increased significantly in the SR (p=0.02) but not the AF cohort, after HUT. During IHG exercise, there was a significant decrease in cardiac BNP release in the AF cohort (p=0.03) but not the SR cohort. CONCLUSION: These data imply in HF, cardiac rhythm influences cardiac BNP release in response to haemodynamic stress. PMID- 16309957 TI - Myocardial regulatory proteins and heart failure. AB - Cardiac troponin T (cTnT) and cardiac troponin I (cTnI) are considered to be the most specific and sensitive biochemical markers of myocardial damage. Troponins have been studied in a wide range of clinical settings, including heart failure; however, there are few data on the role of regulatory proteins in the pathogenesis of heart failure, although a few interesting hypotheses have been proposed. A considerable body of evidence favours the view that alteration of the myocardial thin filament is the primary event leading to defective contractility of the failing myocardium, while the changes in Ca(2+) handling are a compensatory response. A better understanding of the role of regulatory proteins under different physiological and pathological conditions could lead to new therapeutic approaches in heart failure. Recently, calcium sensitisation has been proposed as a novel method by which cardiac performance may be enhanced via an increase in the affinity of troponin C for calcium but without affecting intracellular calcium concentration. To date, the only calcium sensitizer used in clinical practice is levosimendan. PMID- 16309958 TI - An efficacy, safety, and dose-response study of Ramelteon in patients with chronic primary insomnia. AB - BACKGROUND AND PURPOSE: To evaluate the efficacy, safety, and dose response of Ramelteon, a novel highly selective MT1/MT2 receptor agonist, in patients with chronic primary insomnia. PATIENTS AND METHODS: A randomized, multicenter, double blind, placebo-controlled, five-period crossover study design was performed. A total of 107 patients, aged 18-64 years, were randomized into a dosing sequence that included 4, 8, 16, and 32 mg of Ramelteon and placebo. Patients received all five treatments, with a 5- to 12-day washout period between treatments, and served as their own controls. Medication was administered 30 min before habitual bedtime and polysomnographic monitoring. Next-day residual effects were assessed with two visual analog scales (mood and feeling), digit symbol substitution test (DSST), word-list memory tests (immediate recall and delayed recall), and a post sleep questionnaire that ascertained patients' alertness and ability to concentrate. RESULTS: All tested doses of Ramelteon resulted in statistically significant reductions in latency to persistent sleep (LPS) and increases in total sleep time (TST). No next-day residual effects were apparent at any dose, as compared with placebo. There were no differences in the number or type of adverse events between any active treatment and placebo group. The most commonly reported adverse events were headache, somnolence, and sore throat. CONCLUSIONS: Ramelteon demonstrated a statistically significant reduction in LPS and a statistically significant increase in TST, with no apparent next-day residual effects, in patients with chronic primary insomnia. PMID- 16309959 TI - Sleep disturbances and teacher ratings of school achievement and temperament in children. AB - BACKGROUND: The current study examined the relationships between academic achievement, sleep, temperament and demographic-historical data in school-age children. METHODS: Teachers were asked to fill out the teacher temperament questionnaire and a form for school achievement, while mothers filled out a demographic-historical form and the sleep disturbance scale for children (SDSC), a 26-item questionnaire that consisted of six factors: difficulty in initiating and maintaining sleep (DIMS), sleep breathing disorders (SBD), arousal disorders (DA), sleep-wake transition disorders (SWTD), disorders of excessive somnolence (DOES), sleep hyperhydrosis (SHY). From a sample of 380 school children, 264 (70%) were suitable for the analyses (141 M and 123 F aged 8-11 years, mean 9.6 years). A school achievement index (SAI) was derived, summing up four items (reading ability, reading comprehension, mathematics, executive ability) of the teacher form for school achievement. RESULTS: SAI mean for the total sample was 11.1 (SD=2.8). A significant gender difference was found: females have higher SAI than males. The analysis of the demographic-historical form showed that only two factors appeared to affect SAI score: enuresis and the low educational level of the mother. The pattern of correlations showed that (a) the SAI was negatively related to the SDSC total score, in particular to DIMS and DOES sub-factors, and (b) the SAI was significantly and positively correlated with the temperamental traits of task-orientation and personal-social flexibility. A multiple stepwise regression analysis showed that the temperamental traits task-orientation and personal-social flexibility are the most predictive factors for SAI while the mother's educational level and the SDSC total score, although they contributed significantly to the prediction of SAI, accounted for only a small portion of variance. CONCLUSIONS: Temperament and sleep are important factors influencing school achievement, and their assessment could help to identify children at risk regarding school achievement. PMID- 16309960 TI - Limitation of the palatal surgery in OSA patients. PMID- 16309961 TI - Intermittent hypoxia causes REM sleep deficits and decreases EEG delta power in NREM sleep in the C57BL/6J mouse. AB - BACKGROUND AND PURPOSE: Obstructive sleep apnea (OSA) severely impairs sleep architecture. We hypothesized that both intermittent hypoxia (IH) and non-hypoxic arousals of OSA result in significant disruption of non-rapid eye movement sleep (NREMS) and rapid eye movement sleep (REMS). PATIENTS AND METHODS: Polysomnography was performed in C57BL/6J mice (n=5) exposed to IH (cycling of FIO2 from 20.9 to 5.0%) or sleep fragmentation (SF: high flow air blasts) throughout the 12-h light phase over 5 consecutive days. RESULTS: Both IH and SF induced arousals from sleep. On Day 1 of exposure, total NREMS during the light phase decreased comparably during IH (44.1+/-7.8%/12h, P<0.05) and SF (43.7+/ 3.3%/12h, P<0.05) but returned to baseline levels of 62.0+/-7.8%/12h by Day 5 of exposure under both conditions. During IH, however, the electroencephalographic (EEG) delta power of NREMS remained impaired throughout the 5-day period of IH with a nadir of 65.4+/-5.6% relative to baseline (P=0.01), and REMS was effectively abolished during the light phase. In contrast, SF did not cause a significant reduction in either EEG delta power or REMS during the light phase. CONCLUSIONS: Thus, hypoxic exposure, but not arousals, caused overall deficits in the EEG delta power of NREMS and marked deficits in the total amount of REMS. We propose that hypoxic arousals may have a more severe impact on sleep architecture in patients with OSA than non-hypoxic arousals. PMID- 16309962 TI - Oxygenation in the artery during nap sleep. PMID- 16309963 TI - Obstructive sleep apnea syndrome: a potential cause of lower airway obstruction in cystic fibrosis. AB - A six-year-old healthy female with cystic fibrosis (CF) and pancreatic sufficiency presented with cough, weight loss, and lung function decline. Further history suggested obstructive sleep apnea, and nocturnal polysomnography (NPSG) confirmed this. Adenotonsillectomy resulted in resolution of clinical symptoms with return of normal lung function. This case establishes that obstructive sleep apnea syndrome (OSAS) may be a potential cause of lower airway inflammation and resulting weight loss in the young CF population. PMID- 16309964 TI - CARD games between virus and host get a new player. AB - A growing family of cellular proteins encoding the caspase activation and recruitment domain (CARD) has a crucial role in immunity by sensing virus infection and signaling antiviral immune defenses. Four independent studies have identified a novel CARD-containing protein, variously called IPS-1, MAVS, VISA and Cardif, which is an essential signaling adaptor of the host defense mediating CARD-CARD interactions with retinoic acid inducible gene-I (RIG-I) and melanoma differentiation-associated gene 5 (MDAS), sensors of virus infection. Disruption of this novel signaling pathway by hepatitis C virus (HCV) might provide a foundation for viral persistence. PMID- 16309965 TI - Polysaccharide-containing conjugate vaccines for fungal diseases. AB - The recognition that antibodies are effective against fungal pathogens has spawned interest in developing vaccines that elicit antibody-mediated protection. Recently, a novel polysaccharide-protein conjugate vaccine that uses the algal antigen laminarin was shown to elicit antibodies to beta-glucan in fungal cell walls and to mediate protection against both experimental candidiasis and aspergillosis. Remarkably, vaccine-induced antibodies manifested direct antifungal effects, suggesting that vaccine efficacy might not require cellular or other components of the immune system. The description of a vaccine that could protect against various fungal pathogens opens exciting new dimensions in the search for approaches to control fungal diseases. PMID- 16309966 TI - A computerised guidance tree (decision aid) for hypertension, based on decision analysis: development and preliminary evaluation. AB - This paper discusses the piloting of a computerised decision aid that provides individualised information about hypertension to patients. The program is based on decision analysis, using decision trees as a way of structuring information. It incorporates the Framingham risk equation to assess a users' risk of coronary artery disease, together with a detailed assessment of the patient's current lifestyle and their willingness to change behaviour. Users of the program can decide how much or how little information they access. The program assesses individual's preferences for different treatment outcomes, before providing them with guidance on what might be the best treatment option for them. The program was evaluated by 10 patients with a diagnosis of mild to moderate hypertension and 8 health care professionals. Overall, both health care professionals and patients assessed the program positively. The use of a decision aid based on decision analysis may be a useful way of providing information to patients in order to promote shared decision making. PMID- 16309967 TI - Optimizing plasmid-based gene transfer for investigating skeletal muscle structure and function. AB - Intramuscular injection of naked plasmid DNA is a less cytotoxic alternative to viral vectors for delivering genetic material to skeletal muscle in vivo. However, the low efficiency of plasmid-based gene transfer limits its potential therapeutic efficacy and/or its use for many experimental applications. Current strategies to enhance transfection efficiency (i.e., electroporation) can cause significant muscle damage, confounding physiological assessments such as muscle contractility. Optimizing protocols to limit damage is critical for accurate physiological, biochemical, and molecular measurements. Following extensive testing, we developed an electroporation protocol that enhances transfection efficiency in skeletal muscles without causing muscle damage. Pretreating mouse tibialis anterior muscles with hyaluronidase and electroporation at 75 V/cm (using 50% vol/vol saline as a vehicle for plasmid DNA) resulted in 22 +/- 5% of the muscle fibers expressing a reporter gene. This protocol did not compromise contractile function of skeletal muscles assessed at both the intact (whole) muscle and the cellular (single fiber) level. Furthermore, ectopic expression of insulin-like growth factor I to levels that induced muscle fiber hypertrophy without causing tissue damage or compromising muscle function highlights the therapeutic potential of these methods for myopathies, muscle wasting disorders, and other pathophysiologic conditions. PMID- 16309969 TI - Purification of a lysozyme from skin secretions of Bufo andrewsi. AB - A novel toad lysozyme (named BA-lysozyme) was purified from skin secretions of Bufo andrewsi by a three-step chromatography procedure. BA-lysozyme is a single chain protein and the apparent molecular weight is about 15 kDa as judged by SDS PAGE. The specific lytic activity against Micrococcus lysodeikticus of BA lysozyme is 2.7 x 10(5) units/mg, indicating that it is a potent lysozyme. It displayed potent bactericidal activity against Staphylococcus aureus and Escherichia coli with minimal inhibitory concentrations (MIC) of 1 and 8 microM, respectively. The deduced primary structure of BA-lysozyme from cloned cDNA was confirmed by N-terminal sequencing and peptide mass fingerprinting. Its amino acid sequence shares 56.5% identity with that of chicken egg-white lysozyme. Phylogenetic analysis indicates that B. andrewsi lysozyme is closely related to that of turtle. This is the first report on the isolation and primary structure determination of amphibian lysozyme. PMID- 16309971 TI - Lysophosphatidylcholine-induced elevation of asymmetric dimethylarginine level by the NADPH oxidase pathway in endothelial cells. AB - Recent studies suggested that endothelium is a main source of reactive oxygen species (ROS) and the major source was via NADPH oxidase pathway. Various stimuli including lysophosphatidylcholine (LPC), a major component of oxidized low density lipoprotein (ox-LDL), can enhance the activity of NADPH oxidase and lead to a marked ROS generation. Asymmetric dimethylarginine (ADMA) is an endogenous nitric oxide (NO) synthase (NOS) inhibitor, which is synthesized by protein arginine methyltransferase I (PRMT I) and degraded by dimethylarginine dimethylaminohydrolase (DDAH) in endothelial cells. Much evidence showed that ADMA was closely related to endothelial dysfunction. Our previous study showed that LPC elevated ADMA level in endothelial cells via increasing oxidative stress, but the precise cellular mechanism is not defined yet. The present study was to explore the mechanism of NADPH oxidase in LPC-induced elevation of ADMA. In LPC-treated endothelial cells, the ROS production, cell viability, ADMA and NO levels, the activity of DDAH and expression of PRMT I were detected. Treatment with LPC (10 microg/ml) for 24 h markedly increased intracellular ROS production, the expression of PRMT I, level of ADMA, decreased the concentration of NO and the activity of DDAH. These effects were attenuated by diphenyliodonium, the NADPH oxidase inhibitor. In summary, the present results suggested that LPC induced elevation of ADMA was due to reduction of DDAH activity and the up regulation of PRMT expression by stimulation of ROS production via NADPH oxidase pathway. PMID- 16309972 TI - Detection of dopamine in the pharmacy with a carbon nanotube paste electrode using voltammetry. AB - A simply prepared DNA immobilized on a carbon nanotube paste electrode (CNTPE) was utilized to monitor dopamine ion concentration using the cyclic voltammetry (CV) and square-wave (SW) stripping voltammetry methods. The optimum analytical conditions were sought. The result obtained was a very low detection limit compared to other common voltammetry methods. The optimal parameters were found to be as follows: 3.5 pH, 0.48 V SW amplitude, 71 Hz frequency, 5 s accumulation time, 0.01 V increment potential, and -1.3 V (anodic-*-) and 1.2 V (cathodic-o-) accumulation potentials. Given these conditions, the linear working range was observed to be within 0.01-0.11 microg L(-1) (SW anodic and CV). The analytical detection limit was determined to be SW anodic and CV: 4.0 microg L(-1) (2.1 x 10(-11) mol L(-1)) dopamin, and the relative standard deviation at the dopamine concentration of SW anodic 0.05 microg L(-1) was 0.02% (n=15) at the optimum conditions. PMID- 16309973 TI - Impaired DNA double strand break repair in cells from Nijmegen breakage syndrome patients. AB - Nijmegen breakage syndrome, caused by mutations in the NBS1 gene, is an autosomal recessive chromosomal instability disorder characterized by cancer predisposition. Cells isolated from Nijmegen breakage syndrome patients display increased levels of spontaneous chromosome aberrations and sensitivity to ionizing radiation. Here, we have investigated DNA double strand break repair pathways of homologous recombination, including single strand annealing, and non homologous end-joining in Nijmegen breakage syndrome patient cells. We used recently developed GFP-YFP-based plasmid substrates to measure the efficiency of DNA double strand break repair. Both single strand annealing and non-homologous end-joining processes were markedly impaired in NBS1-deficient cells, and repair proficiency was restored upon re-introduction of full length NBS1 cDNA. Despite the observed defects in the repair efficiency, no apparent differences in homologous recombination or non-homologous end-joining effector proteins RAD51, KU70, KU86, or DNA-PK(CS) were observed. Furthermore, comparative analysis of junction sequences of plasmids recovered from NBS1-deficient and NBS1 complemented cells revealed increased dependence on microhomology-mediated end joining DNA repair process in NBS1-complemented cells. PMID- 16309974 TI - Diuretic-induced dehydration/hypovolemia inhibits thermal panting in rabbits. AB - Respiratory and thermoregulatory responses to hyperthermia during isosmotic dehydration/hypovolemia were studied in 17 anaesthetized adult rabbits divided into two groups: normovolemic group (NV; n=10) and hypovolemic group (HV; n=7). Hypovolemia/isosmotic dehydration (a decrease in plasma volume by 16.1+/-1.2%) was induced by furosemide (5 mg kg(-1) i.v.). During hyperthermia (the rise in body temperature to 42 degrees C by a gradual body surface heating), the HV rabbits had lower (P<0.05) respiratory frequency and higher (P<0.05) tidal volume than the NV animals. The panting was absent in the HV rabbits at the BT of 42 degrees C, unlike the NV animals. The lower respiratory frequency and the absence of panting during exogenous hyperthermia in dehydrated animals are present not only during hyperosmotic dehydration induced by water deprivation [Doris, P.A., Baker, M.A., 1981. Hypothalamic control of thermoregulation during dehydration. Brain Res. 206 (1), 219-222], but they also occur in the furosemide-induced isosmotic dehydration/hypovolemia. PMID- 16309975 TI - Validity criteria and comparison of analytical methods of flow-independent exhaled NO parameters. AB - The objective was to assess both validity and comparability of multiple constant (MCF, mainly performed) and dynamically changing (DCF, new method) flow analyses calculating alveolar concentration (Calv(NO)), maximum conducting airway flux (J'aw(NO)) and airway diffusing capacity (Daw(NO)) of exhaled NO (FE(NO)). (Calv(NO), J'aw(NO))(R) where R is the correlation coefficient of the linear regression between NO output and expiratory flow rate (MCF) and (Calv(NO), J'aw(NO), Daw(NO))(Delta100) where Delta100 is the ratio ([observed-predicted FE(NO)]/observed FE(NO)) at 100 ml/s (DCF) were assessed in 18 healthy subjects (10 atopic). MCF demonstrated a linear relationship (R > or = 0.80) between NO output and expiratory flow in 15/18 subjects. DCF was valid (Delta100 < or = 30%) in 12/18 subjects. A good agreement between MCF and DCF was evidenced in the nine subjects with R > or = 0.80 and Delta100 < or = 30%. Failure of validity criteria was mainly observed in atopic subjects. In conclusion, when validity criteria are satisfied, the new DCF method similarly characterizes NO exchange parameters than MCF approach. PMID- 16309976 TI - Possible modulation of the mouse respiratory rhythm generator by A1/C1 neurones. AB - Although compelling evidence exist that the respiratory rhythm generator is modulated by endogenous noradrenaline released from pontine A5 and A6 neurones, we examined whether medullary catecholaminergic neurones also participated in respiratory rhythm modulation. Experiments were performed in neonatal (postnatal days 0-6, P0-P6) and young mice (P14-P18) using "en bloc" medullary preparations (pons resected) and transverse medullary slices. In "en bloc" preparations, blockade of medullary alpha2 adrenoceptors with yohimbine and activation of catecholamine biosynthesis with L-tyrosine significantly depresses and facilitates the respiratory rhythm, respectively. In slices from neonatal and young mice, blockade of medullary alpha2 adrenoceptors also depressed the respiratory rhythm. Yohimbine local applications and lesion-ablation experiments of the dorsal medulla revealed implication of A1/C1 neurones in the yohimbine depressing effect. Although the mechanisms responsible for the yohimbine depressing effect remain to be elucidated, our in vitro results in neonatal and young mice suggest that endogenous catecholamines released from A1/C1 neurones participate in respiratory rhythm modulation via medullary alpha2 adrenoceptors. PMID- 16309977 TI - A physiological model of the release of gas bubbles from crevices under decompression. AB - Moving bubbles have been observed in the blood during or after decompression using ultrasonic techniques. It has been proposed that these may grow from nuclei housed on the blood vessel wall. One candidate for bubble nucleation is hydrophobic crevices. This work explores the growth of gas pockets that might exist in conical crevices and the release of bubbles from these crevices under decompression. An existing dynamic mathematical model for the stability of gas pockets in crevices [Chappell, M.A., Payne, S.J., in press. A physiological model of gas pockets in crevices and their behavior under compression. Respir. Physiol. Neurobiol.] is extended to include the behavior as the gas pocket reaches the crevice mouth and bubbles seed into the bloodstream. The behavior of the crevice bubble is explored for a single inert gas, both alone and with metabolic gases included. It was found that the presence of metabolic gases has a significant effect on the behavior under decompression and that this appears to be due to the high diffusivity of these gases. PMID- 16309978 TI - Influence of hyperoxia on pulmonary O2 uptake kinetics following the onset of exercise in humans. AB - The purpose of this study was to examine the influence of hyperoxic gas (50% O2 in N2) inspiration on pulmonary oxygen uptake (V(O2)) kinetics during step transitions to moderate, severe and supra-maximal intensity cycle exercise. Seven healthy male subjects completed repeat transitions to moderate (90% of the gas exchange threshold, GET), severe (70% of the difference between the GET and V(O2) peak) and supra-maximal (105% V(O2) peak) intensity work rates while breathing either normoxic (N) or hyperoxic (H) gas before and during exercise. Hyperoxia had no significant effect on the Phase II V(O2) time constant during moderate (N: 28+/-3s versus H: 31+/-7s), severe (N: 32+/-9s versus H: 33+/-6s) or supra maximal (N: 37+/-9s versus H: 37+/-9s) exercise. Hyperoxia resulted in a 45% reduction in the amplitude of the V(O2) slow component during severe exercise (N: 0.60+/-0.21 L min(-1) versus H: 0.33+/-0.17 L min(-1); P < 0.05) and a 15% extension of time to exhaustion during supra-maximal exercise (N: 173+/-28 s versus H: 198+/-41 s; P < 0.05). These results indicate that the Phase II V(O2) kinetics are not normally constrained by (diffusional) O2 transport limitations during moderate, severe or supra-maximal intensity exercise in young healthy subjects performing upright cycle exercise. PMID- 16309979 TI - Column-switching high-performance liquid chromatographic method for the determination of zaltoprofen in rat plasma. AB - A direct injection column-switching high-performance liquid chromatography (HPLC) method was developed and validated for quantification of zaltoprofen in rat plasma. Following dilution with mobile phase A, i.e. acetonitrile-10mM potassium phosphate buffer (pH 6.8) (12:88, v/v) samples were directly injected to the pre column without sample pre-purification step. After endogenous plasma components were eluted to waste, the system was switched and the analyte was eluted to the trap column. Zaltoprofen was then back-flushed to the analytical column for separation with mobile phase B, i.e. acetonitrile-10mM potassium phosphate buffer (pH 6.8) (35:65, v/v) and quantification with an ultraviolet detector at 230 nm. The calibration curve was linear in the concentration range of 40-5000 ngmL(-1). This method has been fully validated and shown to be specific, accurate and precise. The method is simple, rapid and the sample preparation is minimal and appears to be useful for the pharmacokinetic study of zaltoprofen. PMID- 16309980 TI - A novel, quantitative assay for homocarnosine in cerebrospinal fluid using stable isotope dilution liquid chromatography-tandem mass spectrometry. AB - We describe a rapid and sensitive method for the quantification of homocarnosine in physiological fluids, with particular emphasis on cerebrospinal fluid (CSF). Homocarnosine was quantified as the butyl derivative, with (2)H(2)-l homocarnosine as internal standard. Following deproteinization of CSF samples, supernatants were evaporated to dryness and derivatized with 10% 6M HCl in butanol. Samples were chromatographed on a C(18) column and detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) operating in the multiple reaction monitoring mode. The intra- and inter-assay variations were 4.6 and 10.9%, respectively. Mean recovery of homocarnosine at two concentrations was 105%. The limit of detection in CSF approximated 20 nmol/L. CSF homocarnosine is age dependent and ranges from <0.02 to 10 micromol/L. Our method is applicable to the analysis of CSF derived from patients with heritable defects in the GABA pathway, patients with homocarnosinosis or serum carnosinase deficiency, and should be applicable to other model systems in order to further explore the biological role and significance of homocarnosine in mammalian systems. PMID- 16309981 TI - Analysis of interaction between DNA and Deinococcus radiodurans PprA protein by atomic force microscopy. AB - A DNA repair-promoting protein, PprA, was isolated from a radiation resistant bacterium, Deinococcus radiodurans [I. Narumi, K. Sato, S. Cui, T. Funayama, S. Kitayama, H. Watanabe, PprA: a novel protein from Deinococcus radiodurans that stimulates DNA ligation, Mol. Microbiol. 54 (2004) 278-285]. Despite several studies, however, the function of PprA is not still clear. We used atomic force microscopy (AFM) to elucidate the role of this protein in the DNA repair pathway. In the present study, interaction between the linear DNA and PprA protein was imaged and analyzed by AFM without any fixation or staining. Though both end bound and internally bound PprA was observed, the affinity of the end-bound protein was greater considering the proportion of features of binding analyzed by AFM. In some conditions, looping forms of the DNA-PprA complex were observed. Gel filtration high performance liquid chromatography (HPLC) was also conducted to estimate the molecular weight of this protein. The result of the HPLC analysis suggested that PprA formed multimers in buffer solution without DNA. PMID- 16309982 TI - Proteomic analysis of the venom and characterization of toxins specific for Na+ - and K+ -channels from the Colombian scorpion Tityus pachyurus. AB - The Colombian scorpion Tityus pachyurus is toxic to humans and is capable of producing fatal accidents, but nothing is known about its venom components. This communication reports the separation of at least 57 fractions from the venom by high performance liquid chromatography. From these, at least 104 distinct molecular weight compounds were identified by mass spectrometry analysis. The complete amino acid sequences of three peptides were determined and the partial sequences of three others were also identified. Electrophysiological experiments conducted with ion-channels expressed heterologously on Sf9 cells showed the presence of a potent Shaker B K(+)-channel blocker. This peptide (trivial name Tpa1) contains 23 amino acid residues closely packed by three disulfide bridges with a molecular mass of 2,457 atomic mass units. It is the third member of the sub-family 13, for which the systematic name is proposed to be alpha-KTx13.3. The mice assay showed clearly the presence of toxic peptides to mammals. One of them named Tpa2, containing 65 amino acid residues with molecular mass of 7,522.5 atomic mass units, is stabilized by four disulfide bridges. It was shown to modify the Na(+)-currents of F-11 and TE671 cells in culture, similar to the beta scorpion toxins. These results demonstrate the presence of toxic peptides in the venom of T. pachyurus and confirm that accidents with this species of scorpion should be considered an important human hazard in Colombia. PMID- 16309983 TI - Kinase domain mutation of MLK4 gene is uncommon in gastric and hepatocellular carcinomas. PMID- 16309984 TI - Is there any correlation between 13C-urea breath test values and response to first-line and rescue Helicobacter pylori eradication therapies? AB - AIM: To study if there is a correlation between 13C-urea breath test values prior to treatment and the response to first-line and rescue Helicobacter pylori eradication therapies. METHODS: Six-hundred patients with peptic ulcer or functional dyspepsia infected by H. pylori were prospectively studied. Pre treatment H. pylori infection was established by 13C-urea breath test. Three hundred and twelve patients were treated with first-line eradication regimen, and 288 received a rescue regimen. H. pylori eradication was defined as a negative 13C-urea breath test, 8 weeks after completion of treatment. RESULTS: H. pylori eradication was achieved in 444 patients. No statistically significant differences were demonstrated when mean delta 13C-urea breath test values were compared between patients with eradication success and failure (49.4+/-33 versus 49.2+/-31). Differences in mean pre-treatment delta 13CO2 between patients with eradication success/failure were not demonstrated either when first-line or rescue regimens were prescribed. With the cut-off point of pre-treatment delta 13CO2 set at 35 units, sensitivity and specificity for the prediction of H. pylori eradication success was 43 and 60%. The area under the receiver operating characteristic curve evaluating all the cut-off points of the pre-treatment delta 13CO2 for the diagnosis of H. pylori eradication was 0.5. Finally, delta 13CO2 values did not influence the eradication in the logistic regression model. CONCLUSION: No correlation was observed between 13C-urea breath test values before treatment and the response to first-line and rescue H. pylori eradication therapies. Therefore, we conclude that the quantification of delta 13CO2 prior to treatment is not useful to predict the success or failure of eradicating therapy. PMID- 16309985 TI - RANKL-stimulated osteoclast-like cell formation in vitro is partially dependent on endogenous interleukin-1 production. AB - Receptor activator of NF-kappaB ligand (RANKL) and interleukin-1 (IL-1) individually plays a critical role in the differentiation and activation of osteoclasts in bone. In addition, both RANKL and IL-1 activate similar signal transduction pathways including p38 MAP kinase and c-Jun NH(2) terminal kinase (JNK). We examined if endogenously produced IL-1 influenced osteoclast-like cell (OCL) formation in murine bone marrow and bone marrow monocyte (BMM) cultures that were stimulated with M-CSF and RANKL. RANKL stimulated OCL formation in a dose-dependent manner in bone marrow cultures, and this response was significantly inhibited by IL-1 RA (100 ng/ml), a specific IL-1 antagonist. Interleukin-1 further increased OCL formation in BMM cultures that were treated with M-CSF (30 ng/ml) and RANKL (1, 3, 10 and 30 ng/ml). In addition, BMM cultures from IL-1 type I receptor-deficient mice, which do not respond to IL-1, demonstrated significantly less OCL formation compared to wild-type BMM cultures. We examined the time course and dose response of IL-1alpha protein expression by ELISA in BMM cultures that were treated with or without M-CSF and RANKL. RANKL dose dependently stimulated IL-1alpha protein significantly (up to 46%) in 6-day cultures. The interaction of RANKL and IL-1 on osteoclastogenesis did not appear significantly dependent on prostaglandin synthesis since PGE(2) expression in the conditioned medium of BMM cultures was nearly undetectable and the PGHS-2 specific inhibitor, NS-398, was without effect. We also investigated the effect of IL-1 on p38 MAP kinase and JNK in BMM cultures. The combination of RANKL and IL-1 had additive effects on JNK but not p38 MAP kinase compared to results in cultures treated with RANKL or IL-1 alone. In addition, SP600125, a specific JNK inhibitor, markedly reduced OCL formation in BMM cultures that were treated with RANKL or the combination of RANKL and IL-1. These findings demonstrate that endogenously produced IL-1 augments the response of bone marrow cells to RANKL, and this effect appears mediated by mechanisms that are associated with enhancement of JNK activity. PMID- 16309986 TI - Vitamin D binding protein, bone status and body composition in community-dwelling elderly men. AB - The vitamin D binding protein (DBP) is the major carrier protein for vitamin D metabolites in plasma. Polymorphisms in DBP have been described to be associated with an increased bone fracture risk and diabetes. The present study investigates the influence of both phenotypic and (TAAA)(n)-Alu repeat DBP-polymorphism and DBP-concentration on bone mineral density, body composition, bone turnover- and metabolic markers in a cohort of ambulatory elderly men. We included 211 men (>70 years) in this study. Bone mineral density (BMD) was determined by dual energy X ray absorptiometry. Bone turnover was assessed by measurement of serum osteocalcin, serum and urinary C-terminal telopeptides of type I collagen and urinary deoxypyridinoline, together with 25(OH)-vitamin D and 1,25(OH)(2)-vitamin D concentrations. DBP-phenotypes were determined electrophoretically and the (TAAA)(n)-Alu repeat polymorphism was determined by polymerase chain reaction. Body composition was estimated using bioelectrical impedance analysis, together with handgrip and arm strength, fasting serum glucose and leptin concentrations. No differences in BMD or bone turnover markers among DBP-phenotypes or (TAAA)(n) genotypes were observed in this study. Serum 25(OH)-vitamin D was comparable among DBP-variants and did not relate to DBP-concentrations, whereas 1,25(OH)(2) vitamin D was different among DBP-phenotypes and was correlated positively with DBP-concentrations. DBP-concentrations related positively to body mass index, fat mass, leptin and glucose concentration. The correlation with leptin remained significant after correction for fat mass. Fasting glucose concentrations were different among DBP-phenotypes, whereas no difference was observed between (TAAA)(n)-genotypes. In conclusion, serum 1,25(OH)(2)-vitamin D concentrations are codetermined by DBP-phenotypes and DBP-concentrations. No major effect of DBP polymorphism was demonstrated on BMD, bone turnover markers or body composition. PMID- 16309988 TI - Total knee arthroplasty for profound varus deformity: technique and radiological results in 173 knees with varus of more than 20 degrees. AB - One hundred seventy-three knees in 117 patients with varus deformity exceeding 20 degrees were prospectively evaluated. Using selective posteromedial release, reduction osteotomy of posteromedial tibial flare, and extra-articular tibial osteotomy, with the preservation of medial collateral ligament integrity, mean tibiofemoral angle of 22.7 degrees varus preoperatively (range, 15 degrees-62 degrees) was corrected to 5.3 degrees valgus (range, 2 degrees-9 degrees) postoperatively. Eighty-six percent of the knees were in 4 degrees to 10 degrees valgus postoperatively. Mean Knee Society score improved from 22.8 (range, 0-64) to 91.1 (range, 52-99), and function score from 22.8 (range, 0-64) to 72.1 (range, 5-100) at 2.6 years (range, 2-9 years). Mean femoral component valgus was 95.5 degrees (range, 92 degrees-98 degrees), and mean tibial component valgus was 89.8 degrees (range, 86 degrees-94 degrees). Of 30 grafts for posteromedial tibial defects, 28 were successfully incorporated. No patient reported significant instability. Three knees (1.7%) showed tibial component loosening with recurrence of deformity. Correction of severe varus deformity by the technique reported can successfully restore alignment, pain-free motion, and stability. PMID- 16309989 TI - Acetabular medial wall displacement osteotomy in total hip arthroplasty: a technique to optimize the acetabular reconstruction in acetabular dysplasia. AB - Twenty-six patients (30 hips) who had acetabular dysplasia were operated on by circumferential acetabular medial wall displacement osteotomy to reconstruct the acetabulum during total hip arthroplasty. All patients had cementless acetabular components implanted. The average acetabular component size was 50 mm (range, 44 56 mm). Only 2 hips needed structural bone graft. The mean follow-up period was 22 months (range, 6-32 months). Harris hip score had changed from 47.31 (range, 19-69 points) to 94.69 (range, 85-100 points) postoperatively (P < .01). Using the Ranawat acetabular triangle to determine the optimal hip center of rotation, the postoperative hip biomechanical environment had been improved. Our short-term follow-up suggests this technique is reliable and reproducible and generally avoids the use of bone graft and graft site morbidity. In addition, it allows the use of standard modular cementless components in patients with acetabular dysplasia. PMID- 16309990 TI - Revision total hip arthroplasty in hip joints with metallosis: a single-center experience with 31 cases. AB - Clinical and radiographic findings of metallosis in relation to the clinical outcome of revision total hip arthroplasty were analyzed for 31 hips. Causes of metallosis were dissociation of polyethylene liner in 24 hips, catastrophic wear in 6, and the screw used for stem fixation in 1. Metallosis was grade I in 12 hips, grade II in 7, and grade III in 12. Osteolysis around the acetabular cup was observed in 24 (77.4%) of 31 hips. At the time of final follow-up evaluation, none of the hips showed osteolysis, acetabular cup migration, or any change in inclination. Although metallosis could not be completely eradicated in severe cases, the clinical outcome for these hips was favorable. This study demonstrates that metallosis frequently causes osteolysis and that complete elimination of it is not a prerequisite for the success of revision total hip arthroplasty. PMID- 16309991 TI - Total hip arthroplasty after failed transtrochanteric rotational osteotomy for avascular necrosis of the femoral head. AB - We compared the clinical and radiographic outcome of 15 hips converted to total hip arthroplasty after failed transtrochanteric rotational osteotomy (THA after TRO) for avascular necrosis of the femoral head (ANFH) with that of a matched control group of 16 hips with primary THA for ANFH. The operating time in THA after TRO was significantly longer than that in primary THA. Perioperative blood loss in THA after TRO was significantly more than that in primary THA. Postoperative complications were observed in 4 hips of THA after TRO. The Harris Hip Score, the stability of implants, and survival rates did not significantly differ in both groups. We conclude that TRO did not influence the outcome of secondary THA at short- and middle-term follow-up. PMID- 16309992 TI - Long-term results of total condylar knee arthroplasty. AB - From 1979 to 1984, 126 consecutive Total Condylar (Howmedica, Rutherford, NJ) knee arthroplasties were performed at a hospital in Taiwan, of which 64 knees were available for evaluation. The patients' average weight was 58 kg. The average Hospital for Special Surgery knee score at the latest follow-up was 86 points. Using revision for mechanical failure as an end point, the 20-year overall survival was 91.9%. The survival for the all-polyethylene tibial component was 96.4% and for the metal-backed tibial component was 88.4%. The difference was statistically significant (P < .001). We recommend the use of the more cost-effective and durable all-polyethylene tibial component for a primary cemented total knee arthroplasty, particularly in Asians with a relatively low weight and who are relatively inactive, especially in elderly people. PMID- 16309993 TI - Total knee arthroplasty in stiff and ankylosed knees. AB - A retrospective study was done to evaluate the results of total knee arthroplasty performed on 84 knees in 53 patients with spontaneously ankylosed knees, having a preoperative arc of movement between 0 degrees and 20 degrees (average, 14 degrees). The difficulties encountered in surgical exposure were managed by using various extensile exposure techniques. Follow-up ranged from 4 to 15 years (average, 9 years). The average postoperative arc of movement at final follow-up was 75 degrees, with a significant average gain of 61 degrees (P < .01). The average extensor lag was 6 degrees. Four knees had a residual flexion contractures of 5 degrees or less. Total knee arthroplasty in ankylosed knees can achieve correction of deformity with a significant gain in range of motion and marked improvement in quality of life. PMID- 16309994 TI - Intraobserver errors in obtaining visually selected anatomic landmarks during registration process in nonimage-based navigation-assisted total knee arthroplasty: a cadaveric experiment. AB - This study investigated the intraobserver errors in obtaining visually selected anatomic landmarks that were used in registration process in a nonimage-based computer-assisted total knee replacement (TKR) system. The landmarks studied were center of distal femur, medial and lateral femoral epicondyle, center of proximal tibia, medial malleolus, and lateral malleolus. Repeated registration in the above sequence was done for 100 times by a single surgeon. The maximum combined errors in the mechanical axis of the lower limb were only 1.32 degrees (varus/valgus) in the coronal plane and 4.17 degrees (flexion/extension) in the sagittal plane. The maximum error in transepicondylar axis was 8.2 degrees. The errors using the visual selection of anatomic landmarks for the registration technique of bony landmarks in nonimage-based navigated TKR did not introduce significant error in the mechanical axis of the lower limb in the coronal plane. However, the error in the transepicondylar axis was significant in the "worst case scenario." PMID- 16309995 TI - Preparation of the flexion gap affects the extension gap in total knee arthroplasty. AB - Changes in the extension gap caused by the preparation of the flexion gap were measured in 50 varus osteoarthritic knees undergoing total knee arthroplasty. The extension gap was first prepared and then measured before and after preparation of the flexion gap. The extension gap increased significantly on both the medial and lateral sides (19.3 +/- 0.3 to 22.0 +/- 0.3 and 21.9 +/- 0.3 to 24.0 +/- 0.3 mm, respectively, mean +/- SE, both P < .0001). The increase was greater medially (P = .0014), and in the knees with more varus deformities (> or = 10 degrees, P = .0075). Thus, preparation of the flexion gap resulted in significant widening of the extension gap, especially on the medial side. The medial release should be stepwise and conservative to achieve adequate soft tissue balance. PMID- 16309996 TI - Prognostic factors influencing the functional outcome of total hip arthroplasty for hip infection sequelae. AB - To investigate factors influencing functional prognosis for patients who have undergone total hip arthroplasty for late sequelae of infective arthritis of the hip, we conducted a retrospective analysis of 75 hips. All patients had had pyogenic or tuberculous infection and later underwent total hip arthroplasty. The average follow-up period was 5.8 years (range, 3-9 years). Various clinical and radiographic measures were analyzed with respect to Harris hip scores. Younger age at the time of infection onset, preoperative leg length discrepancy greater than 1 in, fusion, severe femoral hypoplasia, and severe acetabular dysplasia were associated with poorer prognosis. The complication rate, including 1 recurrent infection, was relatively low. Previous infections seemed to predict poorer results mostly because of the consequences of infection for the development of the hip joint, rather than because of infection recurrence. PMID- 16309997 TI - A new plain radiography method using the optimal angle of knee flexion for assessing early degeneration of the knee joint. AB - During total knee arthroplasty for primary osteoarthritis, we measured the area of worn cartilage on the medial femoral condyle in 44 knees. By reference to a lateral femoral radiograph of each knee, the optimal knee flexion angle reflecting the worn area of the femoral condyle-was determined and found to be 19.8 degrees +/- 7.6 degrees. Using this result, a new method for posteroanterior radiography of the weight-bearing knee (20/10 radiography) was devised. We compared the use of our method with Rosenberg's method in 56 varus knees and found that the width of the cartilage space determined by our method was narrower than that determined by Rosenberg's method (t test, <0.001). These results confirmed that 20/10 radiography well reflects the region of worn cartilage in knees with early-stage primary osteoarthritis. PMID- 16309998 TI - Randomized control trial comparing radiographic total knee arthroplasty implant placement using computer navigation versus conventional technique. AB - The aim of this study is to assess the radiological outcome of conventional techniques versus computer-navigated surgery for total knee arthroplasty. Ninety patients with knee arthritis were prospectively randomized into 3 groups: conventional technique: extramedullary (EM) and intramedullary (IM) tibia guide versus computer navigation surgery (CAS). Two surgeons performed all procedures. Standardized long leg coronal and sagittal x-rays were evaluated by a blinded assessor. Our results showed that CAS had greater consistency and accuracy in implant placement. In the coronal view, 93.3% in the CAS group had better outcomes compared with EM (73.4%) and IM (60.0%). In the sagittal axis, 90.0% CAS also had better outcomes compared with EM (63.3%) and IM (76.7%). Computer navigated total knee arthroplasty helps increase accuracy and reduce "outliers" for implant placement. PMID- 16309999 TI - Early failure of smooth hydroxyapatite-coated press-fit acetabular cup--7 years of follow-up. AB - The early results of smooth hydroxyapatite-coated press-fit acetabular cup are comparable to porous-coated press-fit cup, but these results deteriorate dramatically after 5 years. In this study, 66 primary total hip arthroplasties performed between 1994 and 1998 were reviewed. The average follow-up was 6.5 years. Of the 66 arthroplasties, 8 acetabular cups were revised for aseptic loosening presenting as rapid change of cup position. The average duration of symptoms before revision was 3 months. The survival of the cups at 5 and 9 years were 95.4% and 83%, respectively. The rapid clinical deterioration after 5 years may be due to lack of bone ingrowth into the cup making it unable to resist the shearing stress in the acetabulum. Radiolucent line was not an accurate sign for the detection of mechanical failure. PMID- 16310000 TI - Total hip arthroplasty with the use of nonmodular cementless acetabular component. AB - Sixty-seven hips in 63 patients who underwent total hip arthroplasties with the use of the nonmodular cementless acetabular component and alumina-on-polyethylene bearing surface were available for complete clinical and radiographic review at a mean follow-up period of 7 (range 5-9) years. The mean age was 59 years (range 34 75) years. The mean preoperative Harris hip score of 50 points improved to 93 points at final follow-up. One (1.5%) hip required revision for a recurrent dislocation. No component was loose radiographically at final follow-up. The mean linear wear rate was 0.07 (range 0.01-0.23) mm/y. At a mean follow-up of 7 years, there was no aseptic loosening. Further follow-up, however, is necessary to determine the potential advantage of nonmodular acetabular component for the development of pelvic osteolysis. PMID- 16310001 TI - Effects of hydrogen peroxide solutions on artificial hip joint implants. AB - This study was designed to elucidate the erosive effect of hydrogen peroxide solutions on the materials used for total-hip arthroplasty (THA). As test materials, cross-linked polyethylene, Ti-6Al-4V alloy, and thermal sprayed hydroxyapatite (HA) were used. Changes upon soaking in 3% hydrogen peroxide, before soaking, 1 minute after soaking, 10 minutes after soaking, and 180 minutes after soaking were examined. Scanning electron microscope, Fourier transform infrared analysis, and x-ray diffraction were used for this examination. Hydrogen peroxide did not affect polyethylene, although notable changes in the Ti-6Al-4V alloy and HA did occur. These results indicate that caution should also be exercised to minimize erosion of prosthesis consisting of HA and Ti alloy when hydrogen peroxide solutions are used during total-hip arthroplasty. PMID- 16310002 TI - Is high flexion following total knee arthroplasty safe?: evaluation of knee joint loads in the patients during maximal flexion. AB - The purpose of this study was to indicate the mechanical loads and the flexion angle at the knee during rise from maximal flexion following total knee arthroplasty (TKA). Twenty three knees were evaluated using skin marker-based motion analysis system during four different activities of daily living. The average maximum flexion was 90 degrees (34 degrees less than passive flexion) and all subjects required support for their weight to rise from maximal flexion. The external moments and the external forces at the knee during the maximal flexion were smaller than those during the stair descending activity. The results indicate that capable flexion angle for the patients following TKA is approximately 90 degrees which has smaller mechanical loads at the knee than the stair descending activity. PMID- 16310003 TI - Cementation of a metal-inlay polyethylene liner into a stable metal shell in revision total hip arthroplasty. AB - The purpose of this study was to evaluate the results of liner cementation into a stable acetabular shell using a metal-inlay polyethylene liner in 39 revision total hip arthroplasties. After an average of 2.8 years, 1 cemented liner dislodged from its metal shell at a postoperative 4 years. In the other 38 hips, mean Harris hip scores improved from 65 preoperatively to 86.9 at the final follow-up. Eighteen patients were rated as having excellent results, 17 as good, and 3 as fair. There were no changes in cup position and no cases of osteolytic lesion progression around the femoral and acetabular components in the last follow-up radiographs. Metal-inlay polyethylene liner cementation into a stable acetabular shell was found to provide an alternative option with short-term excellent results, and it also offers more liner options, the preservation of bone stock, and lower surgical morbidity. PMID- 16310004 TI - Premature failure of the polyethylene tibial bearing surface of the Interax knee arthroplasty. AB - Thirty-four patients (37 knees) were treated with the Interax knee arthroplasty. There were 31 female and 3 male patients with a mean age at surgery of 70 years (range, 53-84 years). Premature failure of the polyethylene tibial bearing surface occurred in 9 patients (9 knees). Eight patients of these required a revision operation because of early wearing out of the tibial surface 20 to 38 months after the operation. The failure was due to inadequate implant design in small component and polyethylene gamma irradiated in the air. PMID- 16310005 TI - Total knee arthroplasty in hemophilic arthropathy of the knee. AB - Twenty-five total knee arthroplasties were performed in 21 patients with hemophilia. The mean patient age was 35.8 years and mean follow-up time was 6.2 years. The average preoperative knee score increased from 18.6 points (range, 3 29) to 82.8 points (range, 44-99). The average preoperative knee function score increased from 41.4 points (range, 20-60 points) to 75.8 points (range, 45-95 points). The average preoperative range of motion was 73.4 degrees with an average flexion contracture of 22.6 degrees, whereas the average postoperative range of motion increased to 92.2 degrees with an average flexion contracture of 5.6 degrees. Median consumption of coagulation factor concentrate decreased from 4837 U/month before operation to 1500 U/month 1 year after surgery. The total knee arthroplasty is a useful treatment in severe hemophilic arthropathy to obtain pain relief and functional improvement, and to reduce the need for ongoing treatment using coagulation factor concentrate. PMID- 16310006 TI - Influence of hip position on knee flexion angle in patients undergoing total knee arthroplasty. AB - The influence of hip position on knee flexion angle before total knee arthroplasty (TKA) and that after TKA were investigated and compared. Sixty-six patients (70 knees) with osteoarthritis who were undergoing TKA were examined using lateral radiographs of maximum knee flexion angle with the hip joint at 0 degrees extension and 90 degrees flexion. The postoperative rate of decrease in knee flexion angle caused by hip extension was significantly larger compared with the value before surgery. The preoperative rate of decrease in knee flexion angle caused by hip extension showed strong inverse correlation with the preoperative and postoperative knee flexion angle ratio. In conclusion, tightness of the extensor mechanism is present in all knees undergoing TKA and especially has a strong influence on the postoperative flexion angle of the knee. PMID- 16310007 TI - The early results of high-flex total knee arthroplasty: a minimum of 2 years of follow-up. AB - We retrospectively reviewed our first 25 cases of high-flex total knee arthroplasty (TKA) with an average follow-up of 28 months. The results were compared to a matched group of posterior stabilized (PS) TKA. There was no significant difference in Knee Society Scores between the 2 groups. The high-flex group had an averaged 138 degrees of knee flexion, which was significantly higher than the PS group (average, 126 degrees). Eighty percent of patients in high-flex group were able to squat, which was significantly higher than in PS group (32%). In patients with a small bone frame, occasionally, their bone stock of the posterior femoral condyle is inadequate for high-flex TKA. For the patients without special demands, the additional knee flexion from high-flex design made no significant difference on the results. PMID- 16310008 TI - Laceration of femoral vessels by an avulsion fracture fragment of the lesser trochanter after bipolar hemiarthroplasty. AB - Femoral vessel injuries after bipolar hemiarthroplasty have not been reported. The current report describes a case of a dual major vessel (superficial femoral artery and vein) injury associated with an avulsion fracture fragment of the lesser trochanter in a 76-year-old woman who had been treated with bipolar hemiarthroplasty because of a femoral neck fracture. The superficial femoral artery was repaired and the defect of the superficial femoral vein was reconstructed with a Gore-Tex graft (WL Gore and Associates Inc, Flagstaff, Ariz). The clinical result was satisfactory and there was no vascular problem at 1-year follow-up. Early diagnosis of this vascular injury prevents serious complications including gangrene of the injured limb. PMID- 16310009 TI - Mycobacterium farcinogenes infection after total hip arthroplasty. AB - Atypical mycobacterium infection after total hip arthroplasty is a very rare but a potential cause of implant failure. We present the first report of Mycobacterium farcinogenes infection in human beings. Although the treatment of atypical mycobacterium infection after total hip arthroplasty is controversial, we successfully treated the infection in this case, after consultation with a microbiologist regarding infection management, with both surgery and chemotherapy. It is important to maintain a high index of suspicion for atypical mycobacterium infection, particularly when standard culture findings are negative despite strong clinical evidence of prosthesis infection. PMID- 16310010 TI - Development of a computer-aided diagnosis system for a new modality of renal replacement therapy: an integrated approach combining both peritoneal dialysis and hemodialysis. AB - The authors developed a computer-aided diagnosis system that includes a simple clinical test for the chronic renal disease patient who needs an integrated approach that combines both peritoneal dialysis and hemodialysis (PD-HD therapy). In this case study, the system simulated and estimated the dialysis outcome, the ultrafiltration volume and nutritional analysis by employing a pharmacokinetic model, and assessed the peritoneal permeable enhancement that can be a grave complication with peritoneal dialysis. This system requires only a minimum amount of nursing time and may be able to predict the optimal treatment schedule for PD HD therapy and provide therapeutic monitoring in long-term peritoneal dialysis. PMID- 16310011 TI - Genetic algorithms for parameter estimation in mathematical modeling of glucose metabolism. AB - Direct measurement of hormones secretion and kinetics in glucose metabolism is not feasible in the clinical practice, being highly invasive. As their knowledge is important in the diagnosis of metabolic disorders, thanks to mathematical models based on non-invasive tests, estimation of hormones behaviour is obtained. Unfortunately, traditional model estimation can suffer for convergence problems, and it can be strongly dependent on the parameters initial value. To overcome these limitations, Genetic algorithms (GAs) were tested on a group of 49 subjects. The stochastic nature of GAs allowed overcoming the initialization problem. Moreover, GAs significantly improved the accuracy of fit. PMID- 16310012 TI - A neuro-fuzzy inference system for modeling and prediction of heart rate variability in the neuro-intensive care unit. AB - In the neurological intensive care unit (NICU), prediction of impending changes in patient condition would be highly beneficial. In this paper, we employ a neuro fuzzy inference system (NFIS) for short-term prediction of heart rate variability in the NICU. An NFIS was selected because it allows for a "gray-box" approach through which a system identification procedure is used in conjunction with fuzzy modeling. The NFIS is described in detail and is compared to an auto-regressive moving average (ARMA) model for its ability to model both simulated and measured data from NICU patients. We found that the NFIS is capable of predicting changes in heart rate to a reasonable extent, and that the NFIS has both advantages and limitations over the ARMA model. The NFIS may therefore be a reasonable technique to consider for more extensive prediction purposes in ICU settings. PMID- 16310013 TI - Predicting the anti-hypertensive effect of nitrendipine from plasma concentration profiles using artificial neural networks. AB - Nitrendipine is an effective and safe calcium-channel blocker for the treatment of mild to moderate hypertension. The aim of this study is to show that an artificial neural network (ANN) model of the relationship between nitrendipine plasma levels and pharmacodynamic effects can be built and used for pressure-drop prediction after oral administration of the drug in spite of the poor correlation between plasma concentrations and the effect. To achieve the goal, the following steps were taken: evaluation of the quality of the database for training the ANN, definition of the optimal input set for the ANN, and prediction of the diastolic pressure drop using the ANN. The possible consequences of successful ANN modelling are an optimisation of the drug administration regimen, to achieve the best possible effect, as well as optimal drug formulation for drugs with complicated pharmacokinetic/pharmacodynamic relationships. PMID- 16310014 TI - A system of microcalcifications detection and evaluation of the radiologist: comparative study of the three main races in Malaysia. AB - This paper uses wavelets in the detection comparison of breast cancer among the three main races in Malaysia: Chinese, Malays, and Indians followed by a system that evaluates the radiologist's findings over a period of time to gauge the radiologist's skills in confirming breast cancer cases. The db4 wavelet has been utilized to detect microcalcifications in mammogram-digitized images obtained from Malaysian women sample. The wavelet filter's detection evaluation was done by visual inspection by an expert radiologist to confirm the detection results of those pixels that corresponded to microcalcifications. Detection was counted if the wavelet-detected pixels corresponded to the radiologist's identified microcalcification pixels. After the radiologist's detection confirmation a new client-server radiologist recording and evaluation system is designed to evaluate the findings of the radiologist over some period of cancer detection working time. It is a system that records the findings of the Malaysian radiologist for the presence of breast cancer in Malaysian patients and provides a way of registering the progress of detecting breast cancer of the radiologist by tracking certain metric values such as the sensitivity, specificity, and receiver operator curve (ROC). The initial findings suggest that no single race mammograms are easier for wavelets' detections of microcalcifications and for the radiologist confirmation even though for this study the Chinese race samples detection average were a few percentages less than the other two races, namely the Malay and Indian races. PMID- 16310017 TI - Helicobacter pylori infection and surgical disease---part I. PMID- 16310018 TI - Diagnostic indicators of sleep apnea in older women and men: a prospective study. AB - OBJECTIVE: The purpose of the present prospective study of older adults was to (1) explore the nature and frequency of sleep disorder in a sample of self identified, sleepy/tired individuals and (2) compare symptom presentation of women and men who were subsequently diagnosed with sleep apnea/hypopnea. METHOD: Participants were 112 community-based older adults self-identified with daytime sleepiness, fatigue, or insomnia. They underwent medical examination and overnight polysomnographic recording. Sleep quality, daytime sleepiness, fatigue, psychological adjustment, and perceived health were evaluated by self-report measures. RESULTS: Results indicated (1) a very high rate of sleep disorder identified by the self-selection process, (2) a male-to-female ratio of 1.2 to 1 for diagnosed apnea/hypopnea syndrome, (3) similar severe apnea signs and symptoms reported by both men and women, and (4) virtually no differences in psychological adjustment and few perceived differences in health limitations between men and women. (5) Female participants with sleep apnea/hypopnea could not be distinguished from participants with insomnia only on the basis of reported symptom presentation alone. CONCLUSION: Our findings suggest a strategy for identifying individuals for referral to a sleep laboratory. The findings also underline the diagnostic importance of medically unexplained complaints of daytime fatigue or sleepiness in older adults for the diagnosis and, ultimately, the effective management of sleep apnea and its attendant health risks. PMID- 16310019 TI - Applying a stepped-care approach to the treatment of obesity. AB - OBJECTIVE: In a stepped-care (SC) approach to treatment, more intensive interventions are implemented when less intensive interventions prove to be insufficient. It was hypothesized that a behavioral weight loss program with SC (BWLP+SC) would evidence superior treatment outcomes when compared with a BWLP without SC (BWLP). METHODS: Forty-four obese, sedentary adults were randomly assigned to a BWLP+SC [i.e., problem-solving therapy (PST)] or a BWLP. RESULTS: Participants in the BWLP+SC lost significantly more weight and body fat, reported greater physical activity and greater improvements in diet, and were more likely to achieve their within-treatment weight loss goals than BWLP participants. Participants in the BWLP+SC who received PST (BWLP+SC [PST]) also evidenced superior treatment outcomes including superior weight loss maintenance (through 12 months posttreatment) compared with BWLP participants matched on SC eligibility [BWLP (SC matched)]. CONCLUSION: BWLP+SC may improve treatment outcomes and participant motivation to achieve preestablished weight loss goals. PMID- 16310020 TI - The relationship of diet and exercise for weight control and the quality of life gap associated with diabetes. AB - OBJECTIVE: The demanding behavioral changes for weight control to manage diabetes might contribute to lower quality of life (QOL). This research examines whether the demands of diet and exercise contribute to lower QOL among persons with diabetes. METHODS: Data were from the Behavioral Risk Factor Surveillance System (BRFSS), which measures days in the last month for poor physical health, poor mental health, limited activity, pain, depression, stress, poor sleep, and high energy, as well as weight control efforts. RESULTS: Respondents with diabetes averaged more impaired days on every measure of QOL. Neither efforts to control weight nor dieting were related to any measure. Exercise was associated with reductions in impaired days on all measures. CONCLUSIONS: Exercising to control weight had a powerful effect on reducing QOL differences while attempting to maintain or reduce weight, and dieting to do so had no effect. PMID- 16310021 TI - Effect of progressive muscle relaxation in adolescent female bronchial asthma patients: a randomized, double-blind, controlled study. AB - OBJECTIVE: The aim of this study is to examine the efficacy of progressive muscle relaxation (PMR) on change in blood pressure, lung parameters and heart rate in female adolescent asthmatics. METHOD: In a prospective, randomized, double-blind, controlled study, adolescent female asthmatics (n=31) were tested to find out how the systolic blood pressure (SBP), forced expiratory volume in the first second (FEV(1)), peak expiratory flow (PEF) and heart rate change after PMR. The control group (CG; n=30) received a placebo intervention. RESULTS: A significant reduction in SBP and a significant increase in the FEV(1) and PEF were observed after PMR. The heart rate showed a significant increase in the coefficient of variation (CV), root-mean-square of successive differences (RMSSD) and at the high frequency (HF) range, in addition to a significant reduction at the low and middle frequency (LF and MF, respectively) ranges. CONCLUSION: PMR appears to be effective in improvement of blood pressure, lung parameter and heart rate in adolescent female asthmatics. PMID- 16310022 TI - How age influences the expression of narcolepsy. AB - OBJECTIVES: The aim of this study was to investigate the influence of age on the manifestation of narcolepsy symptoms and cognitive difficulties in patients with narcolepsy. METHODS: A total of 321 participants were included in the study: 157 were patients with narcolepsy from two Sleep Disorders Clinics and 164 were control participants. Narcoleptic patients were evaluated and diagnosed at the Sleep Disorders Clinic. All participants were interviewed by telephone using the Sleep-EVAL System. The interview comprised, among else, a detailed evaluation of narcolepsy symptoms and of cognitive difficulties. RESULTS: The first manifestation of the disease appeared early in life for most narcoleptic patients: 54.1% had their first symptom before the age of 20 years. Daytime sleepiness was the first symptom to appear in 65.5% of cases. In narcoleptics 60 years or older, cataplexy was more likely to be the first symptom to appear (47.4%) compared with other narcoleptic patients (21.4%; P<.05). Reported cognitive difficulties (attention-concentration, praxis, delay recall, orientation for persons, temporal orientation, and prospective memory) were higher in narcoleptic patients compared with the controls. The severity of daytime sleepiness and the presence of a major depressive disorder partly explained the cognitive difficulties. However, attention-concentration deficits and difficulties in prospective memory remained significant. Age was unrelated to cognitive difficulties in narcoleptics patients. CONCLUSIONS: The first manifestation of narcolepsy appears early in life. Reported cognitive difficulties are important in narcoleptic patients and are only partly explained by age, severity of daytime sleepiness, and major depressive disorder. PMID- 16310023 TI - Health-related quality of life and health locus of control beliefs among HIV infected treated patients. AB - OBJECTIVE: This study examined the relations between health locus of control (HLOC) beliefs and health-related quality of life (HRQL) in 302 HIV-infected patients enrolled in a French cohort, 44 months (M44) after they began highly active antiretroviral therapy (HAART). METHODS: HLOC beliefs were measured with the Multidimensional Health Locus of Control (MHLOC) scale and HRQL, with the Medical Outcome Study Short-Form Health Survey (MOS-SF-36). RESULTS: Internal HLOC beliefs at the initiation of treatment were associated with both physical HRQL in multivariate analysis, while chance HLOC beliefs on beginning HAART were associated with mental HRQL at M44. CONCLUSION: These findings suggest the importance of considering the psychological characteristics and psychosocial beliefs of patients at the initiation of ARV treatment to optimise the long-term HRQL of HIV-infected patient and to develop adaptive intervention on coping strategies. PMID- 16310024 TI - The effect of psychological stress on symptom severity and perception in patients with gastro-oesophageal reflux. AB - OBJECTIVE: Using an experimental paradigm this investigation explored whether exposure to psychological stress would produce a significant increase in acid reflux episodes or modify subjective perceptions of gastro-oesophageal reflux (GOR) symptoms. METHODS: Forty-two patients presenting with heartburn and acid regurgitation underwent 24-h oesophageal pH monitoring. During the last 90 min of this monitoring period, 21 patients received a psychological stressor, while the remaining participants were randomly assigned to a no-stress control condition. State anxiety and subjective GOR symptom ratings were obtained 1 min pretest, 1 min posttest, and 40 min posttest. Cortisol samples were collected at 10-min intervals. RESULTS: The stressor induced a significant increase in cortisol and state anxiety; however, this was not associated with any increase in reflux. Instead, the experimental group reported a dissociation between objectively measured reflux episodes and subjective symptom ratings. A similar pattern was established for participants who reported greater state anxiety, produced larger cortisol responses, or exhibited certain stress-related personality characteristics. CONCLUSION: The perception of symptoms in the absence of increased reflux when one is stressed may account for low response rates to traditional treatments. This highlights a need to bridge the gap between psychosomatic research and clinical practice to develop more successful GOR therapies. PMID- 16310025 TI - Stress and heartburn. PMID- 16310026 TI - Psychological characteristics of patients with chronic obstructive pulmonary disease: a review. PMID- 16310027 TI - Psychological characteristics of patients with chronic obstructive pulmonary disease: a review. AB - OBJECTIVE: The aim of this study was to review research on psychological characteristics in chronic obstructive pulmonary disease (COPD) patients and to summarize the findings of associations between psychological variables and various aspects of life, illness, and health in COPD patients. METHODS: The following databases were searched: Medline, PsycINFO, and the Cochrane Library. RESULTS: A total of 81 studies, published between 1968 and 2004, was identified. The reviewed studies indicated that patients suffering from COPD have a high prevalence of psychological disorders and may function at a reduced level of efficiency due to neuropsychological impairments. Psychological characteristics also seemed to be associated with functional disability and reduced quality of life. CONCLUSION: Psychological characteristics are important variables that interact with physical symptoms, and they should be taken into account when treating COPD patients. More studies with rigorous methodological designs are needed to describe the role of psychological variables in COPD accurately and reliably. PMID- 16310028 TI - Consultation liaison psychiatry in Turkey. PMID- 16310029 TI - Competencies of the consultation-liaison psychiatrist: a formulation from the general hospital section of the Dutch psychiatric association. PMID- 16310030 TI - Psychosomatic psychiatry: a Spanish proposal for a new dimension. PMID- 16310031 TI - General hospital psychiatry in the Netherlands. PMID- 16310032 TI - Gestational diabetes and the adenosine/L-arginine/nitric oxide (ALANO) pathway in human umbilical vein endothelium. AB - Altered endothelial cell function is a key factor associated with vascular disorders and is critical in the fetal growth and development. Pregnancies affected by diseases such as gestational diabetes are associated with human umbilical vein endothelial dysfunction, a finding that has been associated with a high incidence of vascular complications during the adult life. Limited information is available addressing cellular mechanisms associated with altered human umbilical vein endothelial function in gestational diabetes. One of the key signalling pathways associated with altered vascular physiology is the synthesis of the vasodilator nitric oxide (NO) from the cationic amino acid L-arginine by the endothelium (i.e. the endothelial L-arginine/NO pathway). The activity of this signalling pathway is modulated by D-glucose, adenosine, insulin, and ATP, among other molecules, and is upregulated (transcriptional, post-transcriptional and post-translational levels) in gestational diabetes. This review focuses on the cellular and molecular mechanisms involved with elevated adenosine levels in fetal umbilical vein blood and the endothelial L-arginine/NO pathway activity in gestational diabetes. We suggest that a lower capacity of adenosine transport by the fetal endothelium in gestational diabetes leads to extracellular accumulation of this nucleoside and its higher bio-availability activates endothelial P1 type purinoceptors. A functional association between A2a purinoceptor subtype signalling and the activity of the l-arginine transport mediated by human cationic amino acid transporters and endothelial NO synthase activity (i.e. 'ALANO pathway') is proposed, revealing in part the mechanisms that account for human umbilical vein endothelial cell dysfunction programmed through the development of the fetus in gestational diabetes. PMID- 16310033 TI - Selective distribution and pregnancy-specific expression of DC-SIGN at the maternal-fetal interface in the rhesus macaque: DC-SIGN is a putative marker of the recognition of pregnancy. AB - We performed immunohistochemical analysis of DC-SIGN expression at the maternal fetal interface at different stages of pregnancy in the rhesus monkey. Natural killer cells, monocytes and macrophages were observed in the nonpregnant endometrium, particularly in the luteal phase, and were increased in pregnant endometrium. No DC-SIGN+ cells were observed in the nonpregnant uterus. We observed decidual DC-SIGN+ cells within 1 week of implantation, and they increased in number during the first 5 weeks of gestation. DC-SIGN+ cells showed a clear differential distribution in the decidua in the first 2 weeks of pregnancy, being found only adjacent to the implantation site, in marked contrast to the widespread distribution of CD68+ macrophages and CD56+ NK cells throughout the endometrium. DC-SIGN+ cells also showed a more dendritic morphology than the general CD68+ cell population, and analysis of serial sections indicated an overlapping but not identical localization of these markers. Mature dendritic cells could not be detected as judged by total absence of immunostaining for CD83, CD86, DEC-205, or CD1a. DC-SIGN+ cells were defined as MHC class II+ and CD14+ by flow cytometry. We conclude that DC-SIGN expression is an early response by the primate maternal immune system to the implanting embryo. The selectively distributed population of DC-SIGN+ decidual leukocytes may represent a morphologically and phenotypically distinct subpopulation of decidual macrophages of early pregnancy that could contribute to the establishment of maternal-fetal immune tolerance. PMID- 16310034 TI - Interstitial trophoblast invasion in the decidua and mesometrial triangle during the last third of pregnancy in the rat. AB - Like other species with haemochorial placentation, pregnant rats show marked invasion of the uterine wall by trophoblast. While an endovascular pathway of invasion has been recognized for a long time, only recently, by application of cytokeratin immunostaining, the existence of an interstitial pathway of invasion has been established. Interstitial invasion is mainly effected by glycogen cell like trophoblast arising from glycogen cell islands of the trophospongium opening up into the decidua, and from glycogen cell sheaths surrounding the intraplacental maternal arterial channels which are connected with the spiral arteries in decidua and mesometrial triangle. Quantitative evaluation of interstitial invasion in both maternal compartments was carried out on days 15 21, using PAS staining and cytokeratin and alpha-actin immunostaining for detecting trophoblast and defining maternal tissue compartments. Measurements of compartment size, cytokeratin-positive areas and invasion extent were performed using the KS400 image analysis system. A distinct pattern of interstitial trophoblast invasion emerged, starting from central decidual areas around the maternal arterial channels, and mushrooming into the mesometrial triangle reaching a peak at day 18, followed by gradual regression of the invaded areas. These measurements may serve as a basis for further experiments to evaluate factors which may influence the depth of trophoblast invasion. PMID- 16310035 TI - Maternal cholestasis induces placental oxidative stress and apoptosis. Protective effect of ursodeoxycholic acid. AB - We have investigated whether maternal obstructive cholestasis during pregnancy (OCP) causes oxidative stress and apoptosis in rat placenta and whether treatment with ursodeoxycholic acid (UDCA, i.g., 60 microg/100 g b.wt./day, following complete biliary obstruction on day 14 of pregnancy) has protective effects on this organ. In rats with OCP, increased (15-fold) serum bile acid concentrations (BAs) together with signs of placental oxidative stress (lipid peroxidation and protein carbonylation) were found. The latter were partly prevented by UDCA, even though hypercholanemia was not corrected. Some elements of the antioxidant system (total glutathione content, GSH/GSSG ratio and catalase, glutathione peroxidase, and glutathione-S-transferase--but not glutathione reductase--activities) were impaired in placentas from the OCP group. UDCA treatment partly prevented changes in the antioxidant system. OCP induced an increase in Bax-alpha/Bcl-2 mRNA ratio, as determined by real-time quantitative PCR, suggesting enhanced susceptibility to apoptosis activation through the mitochondria-mediated pathway. Accordingly, the activity of caspase-3, but not caspase-8, was increased in OCP placentas, in which DNA-ladder analysis and TUNEL confirmed the existence of apoptosis. UDCA prevented changes in the Bax-alpha/Bcl-2 mRNA ratio and caspase-3 activity. In conclusion, OCP causes oxidative stress and apoptosis in rat placenta, which can be prevented by treatment with UDCA. PMID- 16310036 TI - Reactivity of human placental chorionic plate vessels is modified by level of oxygenation: differences between arteries and veins. AB - Normal fetal development in utero is dependent upon adequate perfusion of the placental vasculature, yet how fetoplacental blood flow is matched to maternal blood flow is unknown. In the perfused placental cotyledon in vitro, reduced oxygenation promotes vasoconstriction, which may act to direct blood in fetoplacental vessels to effectively perfused regions of the intervillus space. We aimed to demonstrate that oxygen tension could directly modify placental chorionic plate vessel vasoreactivity. Small arteries and veins from the chorionic plate were dissected from biopsies from term placentae of uncomplicated pregnancies and studied using parallel wire myography. Chorionic artery and vein vasoconstriction in 20%, 7% and 2% oxygen was assessed utilizing the thromboxane mimetic U46619. Reduced oxygenation increased arterial maximal active effective pressure production and sensitivity to U46619. This effect was cyclo-oxygenase independent. In veins, modified oxygenation did not alter vasoconstriction. Vasodilatation in response to the NO donor sodium nitroprusside was increased in lowered oxygenation in veins but not in arteries. We suggest that modified oxygenation may play a role in the control of the blood flow in the fetoplacental circulation. PMID- 16310037 TI - Effects of chronic hypoxia in vivo on the expression of human placental glucose transporters. AB - Birth weight is reduced and the risk of preeclampsia is increased in human high altitude pregnancies. There has been little work to determine whether hypoxia acts directly to reduce fetal growth (e.g. reduced blood flow and oxygen delivery), or via changes in functional capacities such as nutrient transport. We therefore investigated the expression of a primary nutrient transporter, the GLUT1 glucose transporter and two in vitro markers of hypoxia (erythropoietin receptor, EPO-R, and transferrin receptor, TfR) in the syncytial microvillous (MVM) and basal membrane fractions (BMF) of 13 high (3100 m) and 12 low (1600 m) altitude placentas from normal term pregnancies. Birth weight was lower at 3100 m than at 1600 m despite similar gestational age, but none of the infants were clinically designated as fetal growth restriction. EPO-R, TfR and GLUT1 were examined by immunoblotting and maternal circulating erythropoietin and transferrin by ELISA. EPO-R was greater on the MVM (+75%) and BMF (+25%) at 3100 m. TfR was 32% lower on the MVM at 3100 m. GLUT1 was 40% lower in the BMF at 3100 m. Circulating EPO was greater at high altitude, while transferrin was similar, and neither correlated with their membrane receptors. BMF GLUT1 was positively correlated with birth weight at high, but not low altitude. In this in vivo model of chronic placental hypoxia, syncytial EPO-R increased as expected, while nutrient transporters decreased, opposite to what has been observed in vitro. Therefore, hypoxia acts to reduce fetal growth not simply by reducing oxygen delivery, but also by decreasing the density of nutrient transporters. PMID- 16310038 TI - Excess syncytiotrophoblast microparticle shedding is a feature of early-onset pre eclampsia, but not normotensive intrauterine growth restriction. AB - RATIONALE: Syncytiotrophoblast microparticles (STBM) are shed into the maternal circulation in higher amounts in pre-eclampsia compared to normal pregnancy and are believed to be the stimulus for the systemic inflammatory response and endothelial cell damage which characterises the maternal syndrome. The excess shedding of STBM may be caused by hypoxia as a result of poor placentation, which is often a feature of pre-eclampsia. Similar placental pathology occurs in some cases of normotensive intrauterine growth restriction (nIUGR), but in the absence of maternal disease. OBJECTIVE: To examine whether the shedding of STBM in nIUGR occurs to the same extent as in pre-eclampsia. METHODS: A prospective case control study in a tertiary referral centre of: 1) women with early-onset pre eclampsia (EOPET < 34 week), 2) women with late-onset pre-eclampsia (LOPET > or = 34 week), 3) women with nIUGR), 4) matched normal pregnant women (NPC), and 5) non-pregnant women. An ELISA using the antitrophoblast antibody NDOG2 was used to measure STBM levels in peripheral venous plasma. Non-parametric analyses were conducted with statistical significance set at p < 0.05. RESULTS: STBM levels rise during normal pregnancy. EOPET was associated with increased STBM levels (EOPET (median): 41 ng/ml, n = 15) compared with matched normal pregnancy (16 ng/ml, n = 15; Wilcoxon p = 0.005). LOPET (50 ng/ml, n = 10) and nIUGR (18 ng/ml, n = 8) STBM levels did not differ from matched normal pregnancy (36 ng/ml, n = 15, and 36 ng/ml, n = 8, respectively). Background levels in non-pregnant plasma were 0.49 ng/ml, n = 10. CONCLUSIONS: Increased STBM levels were found in EOPET but not in nIUGR providing further evidence for their role in the pathogenesis of the maternal syndrome. PMID- 16310039 TI - The granzyme B inhibitor, PI-9, is differentially expressed during placental development and up-regulated in hydatidiform moles. AB - The intracellular serpin Proteinase Inhibitor-9 (PI-9) is a potent inhibitor of the cytotoxic lymphocyte (CL) proteinase granzyme B, a major effector molecule used by CLs to induce target cell apoptosis. PI-9 is produced by CLs to protect against mis-directed granzyme B. However, PI-9 expression has also been reported in immune privileged tissues. In the present study, cell-specific expression of PI-9 in placental tissue of various gestational ages was examined by immunohistochemistry. PI-9 is highly expressed by the extravillous trophoblasts that have invaded the decidua, and this high expression is maintained throughout pregnancy. Similar levels were also observed in proliferative villous cytotrophoblasts. Syncytial trophoblasts generally do not produce PI-9 to a significant level until the last few weeks of pregnancy. The villous stroma contains mixed populations of PI-9 positive and negative cells throughout pregnancy, with highest expression during the second trimester. Compared to first trimester placentas, syncytial trophoblasts of partial and complete hydatidiform moles showed marked up-regulation of PI-9. Examination of choriocarcinoma cell lines also demonstrated a very high level of PI-9 is produced by these cells, which may provide protection from granzyme B-mediated apoptosis. The cell specific expression of PI-9 in the placenta is consistent with a function in the maintenance of immune privilege, and it is proposed that up-regulated expression of PI-9 in gestational trophoblastic diseases contributes to disease pathogenesis via immune evasion. PMID- 16310040 TI - Differential expression of vascular endothelial growth factor, placenta growth factor and their receptors in placentae from pregnancies complicated by placenta accreta. AB - Placenta accreta is a pregnancy complication characterized by the presence of life-threatening uteroplacental neovascularization. The factors involving its development are unknown. Vascular endothelial growth factor (VEGF), placenta growth factor (PlGF) and their receptors (VEGFR) have important roles in vascular remodeling. We have investigated the differential expression of these proteins in placentae from placenta accreta (cases) and normal placentation (controls). Immunohistochemically, the expression of VEGFR-2 in the syncytiotrophoblast was significantly lower in cases than in controls during both the second and third trimesters (P = 0.005 and 0.002, respectively). However, VEGFR-2 expression in the cytotrophoblastic and extravillous trophoblastic cells and VEGFR-1, -3 and Ki 67 in the trophoblast populations were not significantly different between controls and cases (P > 0.05). Ki-67 immunostaining also showed that endothelial cells of the larger vessels were stained weaker in normal placenta than in placenta accreta. The majority of VEGFR-2 expression, as demonstrated by Western blot or reverse transcription polymerase chain reaction, was consistent with the immunohistochemical findings in the syncytiotrophoblast. Furthermore, enzyme linked immunosorbent assay in the placental lysates showed that the women with placenta accreta demonstrated significantly higher VEGF (P = 0.001) and lower soluble VEGFR-2 (P = 0.015) concentrations than did women with normal pregnancy. PlGF and soluble VEGFR-1 levels did not show any significance in study groups (P > 0.05). These observations suggest that the participation of up-regulated VEGF and down-regulated VEGFR-2 (both membrane-bound and soluble forms) may be associated with the development of placenta accreta. PMID- 16310041 TI - Expression of gap junctional connexins 26, 32 and 43 in bovine placentomes during pregnancy. AB - Gap junctional connexins (Cx) are induced in the endometrium during implantation in rodents, the human receptive window, and in the decidua Cx26 and Cx43 expression increases in response to trophoblast invasion. In contrast, this gap junctional response and decidualization is absent in non-invasive epitheliochorial placentae of pigs and horses. Bovine (syn)epitheliochorial placentation represents an intermediate type of trophoblast invasion, since it is characterized by the continuous migration and fusion of trophoblast giant cells (TGC) with uterine epithelial cells. Therefore the objective of the present study was to investigate the expression of Cx26, Cx32, and Cx43 in placental tissues during bovine pregnancy, to determine if Cx expression patterns correlate with the depth of trophoblast invasion. Cx26, Cx32, and Cx43 proteins were detected by immunohistochemistry and corresponding specific mRNAs were shown by RT-PCR and localized in tissue sections by in situ hybridization. Cx26 protein was detected at the feto-maternal contact interface and as cytoplasmic staining in TGC. Cx26 mRNA was located in maternal epithelium and in TGC. Cx32 protein expression was observed in the maternal epithelium exclusively on the tips of maternal septa, whereas Cx32 mRNA was detected in all maternal epithelial cells and single TGC. Cx43 protein and mRNA were coexpressed in TGC. Cx43 protein was present in maternal septal stroma and to a lesser extent in chorionic villous mesenchyme, while Cx43 mRNA was associated with the vasculature. In the course of gestation, expression of Cx26, Cx32, and Cx43 did not change. In conclusion, the intermediate invasive status of bovine trophoblast is supported by the fact that TGC coexpress Cx26, Cx32, and Cx43, which may be important for trophoblast migration (invasion), and fusion with maternal epithelial cells. Cx32 could be involved in the control of invasion. PMID- 16310042 TI - Placentation in the rock cavy, Kerodon rupestris (Wied). AB - Rock cavies are rodents found in the semi-arid caatinga of Brazil. We studied the structure of the rock cavy placenta by light and transmission electron microscopy. The exchange area of the labyrinth was organized in lobes separated by interlobular areas. The interhaemal barrier was syncytial haemomonochorial. The syncytiotrophoblast had recesses in the basal membrane and some invaginations of the apical membrane, but transtrophoblastic channels could not be found. The interlobular regions comprised syncytiotrophoblast, enclosing maternal venous blood channels, and cytotrophoblast. There was a prominent subplacenta composed of cytotrophoblast and syncytiotrophoblast. Microvilli projected into spaces between the cytotrophoblasts and into lacunae within the syncytiotrophoblast. The yolk sac epithelium exhibited coated pits, endocytotic vesicles and larger vacuoles, consistent with a role in protein uptake from the uterine lumen. Tight junctions between these cells provided a barrier to diffusion by the intercellular route. The reproductive biology of the rock cavy differs from other members of the family, including the guinea pig, but the architecture of the placenta remains remarkably constant. PMID- 16310043 TI - The extent and variability of effects of culture conditions on the secretion of human chorionic gonadotrophin and interleukin-6 by human, term placental explants in culture. AB - Culture of explants derived from third trimester human placenta is used in a range of contexts as an experimental model that retains tissue architecture. This study aimed to explore the variability between, and within, individuals of secretion by explants of human chorionic gonadotrophin (hCG) and interleukin-6 (IL-6). Standard culture medium contained hydrocortisone, insulin, retinoic acid and serum. Under these conditions explants displayed significant differences in the time-course and extent of hCG secretion. Peak hCG secretion varied between 1.19 and 242 mIU/mg protein/h (coefficient of variation (CV) = 111%) and could occur between days 4 and 7 of culture. hCG secretion was more variable if explant protein was < 400 microg. Unadjusted day 7 hCG secretion showed marked variation: intra-placental CV = 15%, inter-placental CV = 86%. When day 7 hCG secretion was standardised by day 6 secretion, intra-placental CV was 6.9%, inter-placental CV was 4.0%. When this standardisation was applied, hCG secretion during day 7 of culture was not affected by removal of hydrocortisone, insulin or serum from the medium or by the addition of tumour necrosis factor-alpha (TNF-alpha). The secretion of IL-6 during day 7 of culture (standardised by taking natural logarithms) was increased markedly by the addition of TNF-alpha but unaltered by removing hydrocortisone, insulin or serum. Thus, we have shown that although variable, secretion by placental explants can be used to investigate how placental tissue adapts to different culture conditions. However, explants of the same protein content may have markedly different secretory properties. PMID- 16310044 TI - Expression of Golf in the rat placenta: Possible implication in olfactory receptor transduction. AB - Olfactory receptors are G-protein coupled receptors and are encoded by an extremely large and diverse family of genes in mammals. There is increasing evidence that olfactory receptors are widely distributed in many organs, suggesting that olfactory receptors do not only recognize airborne odorants but also play important roles in chemotaxis or organ construction in embryo. In this study, we investigated whether olfactory receptors and their transduction molecule, Golf are expressed in the rat placenta. By RT-PCR, we identified 11 different olfactory receptor genes, which are all members of class II, in the rat placenta cDNAs, and our results suggested that particular members of the olfactory receptor gene family might be preferentially expressed in the placenta. By western blot analysis, we demonstrated that Golf protein is expressed in the placenta and its expression levels are developmentally regulated. We found that Golf immunoreactivity is exclusively localized to giant cell trophoblasts and spongiotrophoblast cells. These findings raised a possibility that a particular subset of olfactory receptors might be coupled with Golf and function in giant cell trophoblasts and spongiotrophoblast cells. PMID- 16310045 TI - Human keratinocytes produce the complement inhibitor factor H: synthesis is regulated by interferon-gamma. AB - Locally synthesized complement is believed to play an important role in host defense and inflammation at organ level. In the epidermis, keratinocytes have so far been shown to synthesize two complement components, C3 and factor B. Here, we studied the synthesis of factor H by human keratinocytes. We also studied the regulation of factor H synthesis in keratinocytes by several cytokines, namely IL 1alpha, IL-2, IL-6, TGF-beta1, TNF-alpha and IFN-gamma. Human keratinocytes expressed factor H mRNA and constitutively released small amounts of factor H protein into the culture medium. This release was strongly upregulated by IFN gamma but not by other cytokines tested. Western blot analysis revealed that IFN gamma augments the synthesis of both molecular species, factor H (FH; 155kDa) and factor H-like protein-1 (FHL-1; 45kDa), of factor H. Factor H released in response to IFN-gamma was functionally active. In conclusion, we demonstrate that keratinocytes are capable of synthesizing factor H and that this synthesis is regulated by IFN-gamma. PMID- 16310046 TI - Origins of peripheral B cells in IL-7 receptor-deficient mice. AB - The interleukin 7 (IL-7) signaling pathway is critical for early lymphoid differentiation. We found dramatic perturbations in fetal liver B cell development and confirmed a complete absence of developing B cells in the adult bone marrow in mice lacking the IL-7 receptor alpha (IL-7Ralpha) gene. We show that peripheral B-2 and B-1 cell populations are deficient in IL-7Ralpha-/- mice. B-2 follicular cell and peritoneal B-1 cell percentages are reduced, while B-2 marginal zone cell percentages are increased. A comparison of bone marrow and splenic populations at different ages revealed that the splenic B cell populations seen in adult IL-7Ralpha-/- mice first appear during neonatal development. We have measured N-nucleotide addition at the joints of V(D)J rearrangements in splenic B cells and have used it as a somatic marker to define and separate bone marrow-derived B cells from fetal liver-derived B cells. B cells isolated from the bone marrow and spleen of adult and neonatal IL-7Ralpha deficient mice harbor high levels of N-nucleotide additions similar to those found in equivalent wild-type B cell populations. We conclude that the majority of splenic B cells in IL-7Ralpha-deficient mice originate from the bone marrow and not the fetal liver. PMID- 16310047 TI - AIRE recruits multiple transcriptional components to specific genomic regions through tethering to nuclear matrix. AB - Thymic selection requires that diverse self antigens be presented to developing thymocytes by stromal cells. Consistent with this function, medullary thymic epithelial cells have been shown to express a large number of genes, many of which are tissue restricted. Autoimmune regulator (AIRE) is a nuclear protein, which has recently been identified as a regulator of this process, however, the mechanism by which AIRE functions is not well understood. Here we use a transrepression assay to demonstrate that AIRE interacts with multiple components of the transcription complex including a novel interaction with the UBA domain protein, GBDR1. When AIRE is expressed in cultured human thymic epithelial cells, it tightly associates with nuclear matrix, suggesting that AIRE responsive genes may be localized to specific regions. Using a mathematical approach we have re analyzed an Affymetrix dataset identifying AIRE responsive genes and show that they tend to localize to specific regions of the genome. Together, these data suggest that AIRE regulates gene expression by recruiting components of the transcription complex to specific regions of the genome via interactions with nuclear matrix. PMID- 16310048 TI - Extensive T cell receptor cross-reactivity on structurally diverse haptenated peptides presented by HLA-A2. AB - Previous studies have shown that individual TCRs are able to effectively recognize multiple peptide/MHC complexes that have varying degrees of structural diversity. These TCR cross-reactivities have usually been demonstrated by using peptides that have different amino acid sequences. To further examine the extent to which TCRs can accommodate structurally diverse ligands, we analyzed human TCR cross-reactivity to eight structurally distinct haptens that are coupled to the HLA-A2-binding Tax peptide with a lysine substitution at position 5 (Tax-5K, LLFG[K-hapten]PVYV). The results demonstrate that 71% percent of the haptenated peptide-induced CTL lines could cross-react on at least one other hapten. We compared the effects of HLA-A2 mutants with substitutions at known TCR contact sites for recognition by hapten-cross-reactive CTL. Recognition of the A2 mutants was remarkably similar whether they were presenting the immunizing or the cross reactive peptide, indicating that similar amino acid contacts are made by the TCR during recognition of both complexes. Thus, hapten cross-reactivity is apparently accomplished without major adjustments to the interaction between the TCR and the surface of the HLA-A2 molecule. Collectively, these results suggest that TCRs possess the molecular flexibility to accommodate very structurally diverse ligands while retaining conserved interactions with the surface of the MHC molecule. PMID- 16310049 TI - Effects of 17beta-estradiol on the maturation, nuclear factor kappa B p65 and functions of murine spleen CD11c-positive dendritic cells. AB - Physiological gender differences in immune capabilities are now well recognized and suggest that sex steroid hormones such as estrogens may be involved in the regulation of the immunocompetence. In this paper, CD11c-positive murine spleen dendritic cells (SDCs) were treated with various concentrations of 17beta estradiol (E2) for 24h. The viability, phenotype, nuclear factor kappa B p65 (NF kappaBp65), endocytosis, stimulatory capacity and cytokine expression were analyzed. Our results showed that E2 increased the viability and MHC-II expression but decreased nuclear NF-kappaBp65 level and endocytosis of SDCs. E2 also increased the stimulatory capacity of SDCs from low-dose group but decreased it from middle- and high-dose ones. In addition, E2 increased the intracellular expression of IL-6 and IL-10 in SDCs, but no obvious change appeared in IL-12 and TNF-alpha. These data suggested that E2 might influence the immune responses by changing the viability, maturation, NF-kappaBp65, endocytosis, stimulatory capacity and cytokine expression of SDCs. PMID- 16310050 TI - The catfish liver-expressed antimicrobial peptide 2 (LEAP-2) gene is expressed in a wide range of tissues and developmentally regulated. AB - Antimicrobial peptides (AMPs) are important components of the host's innate immune response against microbial invasion. The cysteine-rich AMPs such as defensin and hepcidin have been extensively studied, but the recently identified cysteine-rich liver-expressed antimicrobial peptide 2 (LEAP-2) has been characterized from only a few organisms. Here we cloned and sequenced the LEAP-2 cDNAs from both Channel catfish and Blue catfish. The LEAP-2 gene from Channel catfish was also sequenced and characterized. Channel catfish LEAP-2 gene consists of two introns and three exons that encode a peptide of 94 amino acids with a 28 amino acid signal peptide and a mature peptide of 41 amino acids. The amino acid sequences and gene organization were conserved between catfish and other organisms. The Channel catfish LEAP-2 gene is expressed in a wide range of tissues except brain and stomach. Its expression is developmentally regulated with no detection of mature mRNA in early stages of development. It appears that the catfish LEAP-2 gene is regulated at the level of splicing; it is constitutively transcribed, but remains unspliced until 6 days after hatching. The expression of LEAP-2 was induced in a tissue-specific manner. Its expression was upregulated in the spleen, but not in the liver and head kidney, after challenge with Edwardsiella ictaluri, the causative agent of enteric septicemia of catfish (ESC). PMID- 16310051 TI - Secretory IgA and mucin-mediated biofilm formation by environmental strains of Escherichia coli: role of type 1 pili. AB - Recent studies suggest the importance of secretory IgA (SIgA) and mucin in the mediation of biofilm formation by commensal bacteria within the mammalian gut. Studies using a variety of strains of Escherichia coli have indicated that the interaction between E. coli and SIgA is dependent on the type 1 pilus. In this study, the importance of the pilus in SIgA-mediated biofilm formation by a laboratory strain (MG1655) and environmental (fecal) strains of E. coli was evaluated. Transient expression of the type 1 pilus by the laboratory strain of E. coli failed to facilitate SIgA-mediated biofilm formation, whereas constitutive expression of the type 1 pilus by the laboratory strain was sufficient. In contrast, transient expression of the type 1 pilus was sufficient to facilitate SIgA-mediated biofilm formation by environmental isolates. The "threshold" for mucin-mediated biofilm formation appeared to be lower than that for SIgA-mediated biofilm formation, perhaps reflecting disparate roles of mucin and SIgA in mediating biofilm formation in the gut. These studies also provide the first procedures for the growth of bacterial biofilms on live epithelial cells in vitro, an important development that may facilitate future studies on the effects of bacterial biofilms on epithelial cells. PMID- 16310052 TI - The novel avian protein, AWAK, contains multiple domains with homology to protease inhibitory modules. AB - We report the purification of a 3.5kDa peptide with antimicrobial activity from the mucosa and epithelial cells of chicken intestine. The peptide contains a pattern of cysteines characteristic of a whey acidic protein (WAP) domain and was identified as the carboxy terminal fragment of a novel 767 amino acid avian protein which has a proposed molecular weight of 81kDa. Using the conserved domain database (CDD) we identified this 81kDa protein to contain multiple amino acid motifs with homology to WAP domains and an amino acid motif with homology to a Kunitz proteinase inhibitor domain. We propose to call this avian protein AWAK (Avian WAP motif containing, Kunitz domain containing). The presence of WAP and Kunitz modules suggests that AWAK has proteinase inhibitor activity. RT-PCR analyses demonstrated expression of the AWAK gene in the chicken intestine. PMID- 16310053 TI - Tropical dermatology: fungal tropical diseases. AB - Fungal infections are common in tropical countries and can have an important impact on public health. Lobomycosis is a common fungal infection in the tropical rain forest of South America, and paracoccidioidomycosis (South American blastomycosis) is a widespread and sometimes severe illness. Penicilliosis marneffei is an opportunistic infection of AIDS patients in southeast Asia. Chromoblastomycosis and mycetomas are causes of morbidity around the world. Sporotrichosis is a worldwide subcutaneous mycosis with a high incidence in tropical countries and is an important illness in immunocompromised patients. Rhinosporidiosis was classed as a fungal infection but is now considered a protistan parasite that belongs to the class Mesomycetozoea. It is included in this review because of its historical classification. In the past, most of these mycoses were restricted to specific geographic areas and natural reservoirs. There are, however, situations in which people from other regions come in contact with the pathogen. A common situation involves an accidental contamination of a traveler or worker who has contact with a tropical mycosis. Even minor trauma to the skin surface or inhalation of the fungal conidia can infect the patient. Thus recognition of the clinical symptoms and the dermatologic findings of the diseases, as well as the geographic distribution of the pathogens, can be critical in diagnosis of the tropical mycoses. This review discusses some of the more common tropical subcutaneous and systemic mycoses, as well as their signs, symptoms, methods of diagnosis, and therapies. LEARNING OBJECTIVE: At the completion of this learning activity, participants should be able to recognize the clinical and histologic presentations of tropical fungal diseases with cutaneous manifestations and be familiar with the appropriate therapies. PMID- 16310054 TI - Elevated 17-hydroxyprogesterone serum values in male patients with acne. AB - BACKGROUND: Androgen excess may provoke or aggravate acne by inducing seborrhea. In women, androgen disorders are frequently suspected when acne is accompanied by hirsutism or irregularities of the menstrual cycle. In men, however, acne may be the only sign of androgen excess. OBJECTIVE: Our aim was to investigate whether male patients with acne display pathologic androgen blood values. METHODS: This case-control study at a university dermatology department with referred and unreferred patients investigated male acne patients (n = 82, consecutive sample) in whom the diagnosis of mild to severe acne was made, as well as a control group of men without acne (n = 38). The main outcome measures were androgen parameters including morning values of testosterone, luteinizing hormone, follicle stimulating hormone, dehydroepiandrosterone sulfate, androstenedione, and 17 hydroxyprogesterone; as well as a corticotropin stimulation test. RESULTS: 17 Hydroxyprogesterone levels were significantly higher (P = .01) in acne patients than in the control group, whereas the other parameters did not differ significantly. In addition, the corticotropin stimulation test revealed abnormal 17-hydroxyprogesterone induction values in 10 of 82 patients. LIMITATIONS: The analysis is limited to a selection of androgen parameters. CONCLUSION: The results suggest that in men irregularities of adrenal steroid metabolism may be a factor contributing to acne. PMID- 16310055 TI - Asymptomatic neurocutaneous melanocytosis in patients with large congenital melanocytic nevi: a study of cases from an Internet-based registry. AB - BACKGROUND: Recent retrospective studies using magnetic resonance imaging (MRI) to screen for neurocutaneous melanocytosis (NCM) among neurologically asymptomatic children with large congenital melanocytic nevi (LCMN) report high prevalence (23-30%) of asymptomatic NCM. We sought to determine prevalence of asymptomatic NCM, and current application of MRI as a screening tool. METHODS: Patients with LCMN from an Internet-based registry answered a questionnaire regarding NCM status. RESULTS: Of 379 patients with LCMN, 26 reportedly had NCM, with 17 reporting neurologic symptoms. Of 186 patients undergoing MRI, 9 reported abnormal findings without neurologic symptoms (4.8%); 80% had LCMN on the posterior axis, whereas 55% had more than 20 satellite nevi. LIMITATIONS: Study data rely on the registry members' self-reported findings and are limited by lack of independent data verification. CONCLUSION: Asymptomatic NCM (determined by MRI) may not be common, with much lower prevalence (4.8%) than previously reported. MRI is widely used for screening patients at risk for NCM, such as patients with LCMN involving the posterior axis and greater than 20 satellite nevi. PMID- 16310056 TI - Nonsteroidal anti-inflammatory drugs and the risk of actinic keratoses and squamous cell cancers of the skin. AB - BACKGROUND: Although animal studies suggest that nonsteroidal anti-inflammatory drugs (NSAIDs), including aspirin, may protect against cutaneous squamous cell carcinoma (SCC) and actinic keratoses (AKs), possible effects on keratinocytic cancers in humans are unknown. OBJECTIVE: We sought to examine the relationship between ingestion of NSAIDs and the risk of SCC and AKs in humans. METHODS: We conducted a case-control study nested within a community-based cohort of 1621 people in southern Queensland, Australia. Eighty-six persons with SCC were compared with 187 age- and sex-matched control subjects randomly selected from within the cohort. NSAID use was captured through face-to-face interviews with study participants, supplemented by color photographs of product packaging. We defined regular use of NSAIDs as consumption of at least two tablets per week (low frequency) or at least 8 tablets per week (high frequency) for at least 1 year. AKs were counted on the face, ears, right hand, and right forearm by a single physician. RESULTS: Patients with SCC were significantly less likely than control subjects to have used any NSAIDs 8 or more times per week for more than 1 year (multivariate odds ratio [OR] 0.07, 95% confidence interval [CI] 0.01-0.71) and to have used full-dose NSAIDs 2 or more times per week for more than 5 years (OR, 0.20; 95% CI, 0.04-0. 96). Among participants without SCC, current regular users of NSAIDs (> or =2 times per week) had significantly lower counts of AKs than nonusers (rate ratio [RR], 0.52; 95% CI, 0.30-0.91). LIMITATIONS: Estimates of NSAID use were based on self-reported data. Statistical power to detect associations was limited by the number of cases with SCC. CONCLUSION: Regular users of NSAIDs appear to have lower risks of SCC and lower counts of AKs than nonusers. PMID- 16310057 TI - Lipomas treated with subcutaneous deoxycholate injections. AB - BACKGROUND: Lipomas are benign neoplasms of mature fat cells. Current treatments are invasive and carry the risk of scarring. Injections of phosphatidylcholine solubilized with deoxycholate, a bile salt, have been used to reduce unwanted accumulations of fat. Recent in vitro and ex vivo investigations indicate that deoxycholate alone causes adipocyte lysis. OBJECTIVE: We sought to report our experience treating lipomas using subcutaneous deoxycholate injections. METHODS: A total of 6 patients presenting with 12 lipomas were treated with intralesional injections of sodium deoxycholate (1.0%, 2.5%, and 5.0%) at intervals of 2 to 20 weeks. Tumor size, cutaneous reactions, and patients' subjective responses were recorded before and after treatment. RESULTS: All lipomas decreased in size (mean area reduction, 75%; range, 37%-100%) as determined by clinical measurement (with ultrasound confirmation in one lipoma) after an average of 2.2 treatments. Several lipomas fragmented or became softer in addition to decreasing in volume. Adverse effects, including transient burning, erythema, and local swelling, were associated with higher deoxycholate concentrations but resolved without intervention. There was no clear association between deoxycholate concentration and efficacy. CONCLUSIONS: Our clinical experience supports our laboratory investigations demonstrating that deoxycholate, rather than phosphatidylcholine, is the active ingredient in subcutaneously injected formulas used to treat adipose tissue. This small series suggests that low concentration deoxycholate may be a relatively safe and effective treatment for small collections of fat. However, controlled clinical trials will be necessary to substantiate these observations. PMID- 16310058 TI - Reflectance-mode confocal microscopy of pigmented skin lesions--improvement in melanoma diagnostic specificity. AB - BACKGROUND: In vivo confocal microscopy enables skin visualization with a quasihistopathologic resolution. OBJECTIVE: We sought to describe confocal features in melanocytic lesions and to evaluate their diagnostic significance for melanoma (MM) identification. METHODS: Thirty seven MMs, 49 acquired nevi, and 16 Spitz/Reed nevi, presenting equivocal clinicodermoscopic aspects were investigated by confocal microscopy. RESULTS: MMs and nevi significantly differed for some aspects. In multivariate analysis, the presence of nonedged dermal papillae, atypical cells, and isolated nucleated cells within dermal papilla, pagetoid cells, widespread pagetoid infiltration, and cerebriform clusters were strongly correlated with MM diagnosis. A receiver operating characteristic curve value of 0.952 was obtained. LIMITATIONS: Spitz/Reed nevi represented a pitfall in confocal diagnosis, owing to the frequent observation of pagetoid infiltration, architectural disarray, and cytologic atypia, and to the impossibility of evaluating cell maturation with depth. CONCLUSION: Characterization of confocal microscopy features of MMs and nevi seems to improve diagnostic accuracy for melanocytic lesions that are difficult to diagnose. PMID- 16310059 TI - Pilot study on the sensitivity and specificity of in vivo reflectance confocal microscopy in the diagnosis of allergic contact dermatitis. AB - The validity, reproducibility, and specificity of patch-testing in the diagnosis of allergic contact dermatitis (ACD) have repeatedly been addressed. In vivo reflectance confocal microscopy (RCM) has been used for real-time evaluation of the histopathologic features of ACD. This pilot study was designed to determine the sensitivity and specificity of RCM in diagnosing ACD in reference to patch testing. Sixteen participants were patch tested with allergens and control substances. Clinical scoring, digital photography, and RCM evaluation were performed at 72 hours, and RCM images were subjected to blinded evaluation. RCM evaluation parameters included stratum corneum (SC) disruption, parakeratosis, stratum spinosum (SS) and stratum granulosum (SG) spongiosis, and exocytosis. Overall, there was high specificity for all RCM features, ranging from 95.8% to 100%. Sensitivity ranged from 51.9% to 96.3%. Significant parameters with high sensitivity and specificity included spongiosis and exocytosis at the level of SS. Logistic regression analysis was performed on significant variables; P values were determined by chi2 analysis. RCM is a promising noninvasive technology for the evaluation of ACD. SC changes are not helpful in the diagnosis of ACD, although the presence of SG spongiosis and SS spongiosis shows high sensitivity in diagnosing ACD. Larger sensitivity and specificity studies are needed and the identification of ACD has to be based on a defined diagnostic algorithm. A limitation of this study is the small sample size; larger sensitivity and specificity studies are needed to confirm these findings. In addition, individual allergens have to be subjected to further evaluations in order to demonstrate the applicability of our findings for other contact allergens. In that regard, RCM may be considered as an adjunctive tool, rather than a substitute, to clinical evaluation. PMID- 16310060 TI - Soft x-ray therapy for cutaneous basal cell and squamous cell carcinomas. AB - BACKGROUND: We have used a schedule for soft x-ray therapy of epithelial malignancies that takes into account the clinically diagnosed tumor involution under treatment. OBJECTIVE: We sought to evaluate the effectiveness of this schedule in terms of cure rate and late ulcerations. METHODS: Patients with 1267 consecutively irradiated (1988-1992) basal cell and squamous cell carcinomas were followed up (average 77 months). RESULTS: The recurrence rate (5.1%) was related to tumor size and thickness and to the time-dose-fractionation factor. The frequency of ulcerations (6.3%) depended on field size, hardness of the x-rays, and in smaller fields (diameter up to 4 cm) on total dose, and time-dose fractionation factor. Of all ulcerations, 82.5 % could be conservatively cured. LIMITATIONS: We have no evidence that our radiation schedule is superior to those published by other authors. CONCLUSION: These results verify the usefulness of soft x-ray therapy for cutaneous epithelial malignancies. PMID- 16310061 TI - Granuloma faciale: a clinicopathologic study of 66 patients. AB - BACKGROUND: Only case reports or studies of small series of patients have focused on granuloma faciale (GF). OBJECTIVE: We sought to describe the clinicopathologic characteristics of GF in a large series of patients. METHODS: We conducted a retrospective analysis of 66 patients and 73 skin specimens. RESULTS: GF mostly presented as reddish plaques or nodules in middle-aged adults. One third of patients had multiple sites involved and 5 patients had extrafacial lesions. A clinical diagnosis of GF was made in only 10 cases; sarcoidosis, lymphoma, lupus, and basal cell carcinoma were the main differential diagnoses. The most frequent histopathologic features were the presence of a grenz zone, neutrophils, and telangiectases. Vascular changes were frequent, although necrotizing vasculitis appeared to be rare. There was often an association of acute and chronic inflammatory patterns, suggesting that GF follows a chronic history with recurrent acute phases, rather than distinct successive acute and chronic stages. LIMITATIONS: In this retrospective study, a reliable analysis of the outcome of patients could not be performed. Results of direct immunofluorescence tests and laboratory investigations were lacking in many cases. CONCLUSION: GF is often clinically misdiagnosed; its morphologic spectrum is broader than usually described and includes a lack of eosinophils, the presence of vascular changes with rare vessel wall necrosis, and associated acute and chronic inflammatory patterns. PMID- 16310062 TI - Infectious occupational exposures in dermatology--a review of risks and prevention measures. I. For all dermatologists. AB - Dermatologists are at risk for exposure to infectious agents from a variety of sources throughout their workday. We review occupational infectious risks from percutaneous exposures, aerosolized infectious particles (eg, with laser surgery and dermabrasion), instrumentation, and cryotherapy. We also discuss current guidelines for management and postexposure prophylaxis of the more common occupational exposures, and conclude with an overview of means to minimize them. PMID- 16310063 TI - Infectious occupational exposures in dermatology--a review of risks and prevention measures. II. The pregnant dermatologist. AB - The number of women of childbearing age (i.e. physicians, registered nurses, physician assistants, nurse practitioners, and medical assistants) working in the field of dermatology is increasing steadily. In part II of our review of the occupational infectious risks in dermatology, we address the special concerns for pregnant health care workers. PMID- 16310064 TI - Cutaneous innervation: form and function. AB - It is useful for dermatologists to know about the innervation of the skin because dysfunction of cutaneous neurons can cause symptoms--such as itching, pain, and paresthesias--that are evaluated by dermatologists. We review the innervation of the skin and update readers about recent neuroscientific discoveries. PMID- 16310065 TI - The indoor UV tanning industry: a review of skin cancer risk, health benefit claims, and regulation. AB - BACKGROUND: Nearly 30 million people tan indoors in the United States annually, including 2.3 million adolescents. Despite increased evidence on the dangers of artificial UV radiation, the popularity of indoor tanning is growing. OBJECTIVES: We aim to assess the following 3 entities: (1) the association of indoor tanning with skin cancer; (2) statements regarding the health benefits of indoor tanning, especially regarding the production of vitamin D; and (3) current regulation of the tanning industry in the United States. METHODS: We conducted a narrative review of the literature. RESULTS: Indoor tanning poses great risks. Studies support the role of artificial UV radiation in cutaneous carcinogenesis. Despite claims by the tanning industry, artificial tanning is not a safe or necessary way to increase systemic vitamin D levels. The National Institutes of Health and the World Health Organization have acknowledged the risks of indoor tanning. Nonetheless, regulations limiting tanning in the United States are surprisingly sparse. LIMITATIONS: Systematic review of the literature was not performed. CONCLUSIONS: Health care providers must increase efforts to warn and educate the public and government about the dangers of UV radiation. PMID- 16310066 TI - Mycophenolate mofetil. PMID- 16310067 TI - Osteopathic medicine and the practice of dermatology: history and current status. An overview of the American Osteopathic College of Dermatology, an affiliate specialty college of the American Osteopathic Association. AB - As the unified training body for osteopathic physicians specializing in dermatology, the American Osteopathic College of Dermatology (AOCD) sponsors 18 dermatology residency programs that integrate academic training (focusing on the core of basic medical sciences) with clinical instruction (emphasizing specialized diagnostic and laboratory techniques and the high-level performance of all dermatologic and surgical procedures) to engender the ability to provide superior, specialized dermatologic care. The board of directors of the American Academy of Dermatology (AAD) recommended passage of a bylaws amendment establishing a category of osteopathic fellow for those osteopathic physicians who are certified by the American Osteopathic Board of Dermatology; however, the AAD membership failed to adopt the amendment by the required two-thirds majority. Constitutional bylaws of both societies stress the importance of the goal of unification and representation of the specialty of dermatology. Continued rapprochement appears to be in the best interest of the AAD and AOCD and may well enhance our efforts to deal with the challenges that face dermatology and medical practice in the 21st century. PMID- 16310068 TI - Defining early mycosis fungoides. AB - This editorial review summarizes the results of 5 meetings sponsored by the International Society for Cutaneous Lymphoma at which the clinicopathologic and ancillary features of early mycosis fungoides were critically examined. Based on this analysis, an algorithm was developed for the diagnosis of early mycosis fungoides involving a holistic integration of clinical, histopathologic, immunopathologic, and molecular biological characteristics. A novel aspect of this algorithm is that it relies on multiple types of criteria rather than just one, for example, histopathology. Before its finalization, the proposed diagnostic algorithm will require validation and possibly further refinement at multiple centers during the next several years. It is anticipated that a more standardized approach to the diagnosis of early mycosis fungoides will have a beneficial impact on the epidemiology, prognostication, treatment, and analysis of clinical trials pertaining to this most common type of cutaneous lymphoma. PMID- 16310069 TI - Medical pearl: Sarcoidosis prognosis. PMID- 16310070 TI - Is pyogenic granuloma associated with Bartonella infection? AB - Lobular capillary hemangioma and bacillary angiomatosis due to Bartonella infection share several clinical and histopathologic characteristics. We sought to determine whether lobular capillary hemangioma is caused by the same agent as bacillary angiomatosis. Forty-five pathology specimens with a histologic diagnosis of lobular capillary hemangioma obtained from patients with the same clinical diagnosis were tested by immunohistochemistry and polymerase chain reaction for the presence of DNA elements of Bartonella spp. None of the 45 lobular capillary hemangioma specimens tested positive for Bartonella spp. We conclude that lobular capillary hemangioma is not associated with Bartonella spp infection. Further research is required to determine the etiologic agent. PMID- 16310071 TI - Increased metastasis and mortality from cutaneous squamous cell carcinoma in patients with chronic lymphocytic leukemia. AB - In patients with chronic lymphocytic leukemia, squamous cell carcinoma behaves aggressively. Our purpose was to compare squamous cell carcinoma metastasis and mortality between patients with chronic lymphocytic leukemia and control subjects. Medical records were assessed retrospectively for 28 patients with chronic lymphocytic leukemia who underwent surgical excision of cutaneous squamous cell carcinoma and for 56 matched control subjects. The rate of metastasis and mortality from cutaneous squamous cell carcinoma were determined on a per-patient basis. Three of 28 patients with chronic lymphocytic leukemia had metastasis and died of metastatic disease. No metastases or deaths occurred among the 56 control subjects. Compared with control subjects, chronic lymphocytic leukemia patients with cutaneous squamous cell carcinoma were more likely to have metastasis (P = .0031) and die of metastasis (P = .0033). No significant association was detected between metastasis and history of chemotherapy administration for chronic lymphocytic leukemia. Among patients with chronic lymphocytic leukemia, surveillance for skin cancer and a decreased threshold for biopsy of suspect lesions are warranted. PMID- 16310072 TI - Pitfalls in the clinical and dermoscopic diagnosis of pigmented actinic keratosis. AB - Pigmented actinic keratosis and melanoma may exhibit overlapping clinical features, thus representing a diagnostic challenge for dermatologists. Although the differentiation between these two entities is traditionally done by histopathology, dermoscopy has been utilized as a useful additional aid for improving the clinical diagnostic accuracy of such pigmented skin lesions. We report the clinical and dermoscopic features of two pigmented actinic keratoses to discuss the difficulties in their preoperative differential diagnosis. PMID- 16310073 TI - Proliferating multicentric reticulohistiocytosis associated with papillary serous carcinoma of the endometrium. AB - Multicentric reticulohistiocytosis (MRH) is a rare histocytic disease characterized by destructive arthritis in association with classic skin findings. Although MRH is not strictly a paraneoplastic disease, one quarter of cases are malignancy related. We report a case of MRH with an initial remission followed by an acute exacerbation several years later heralding the clinical presentation of endometrial carcinoma. During this flareup a skin biopsy specimen revealed a diffuse dermal infiltrate composed of histiocytes with ground-glass cytoplasm and multiple atypical mitoses. Approximately 40% of the cells stained with the proliferation marker Ki-67. Treatment of endometrial carcinoma resulted in improvement of skin and joint symptoms, and a repeat biopsy specimen no longer demonstrated mitotic figures. These findings support a reactive and proliferative cause of MRH. PMID- 16310074 TI - Adverse event reporting for botulinum toxin type A. AB - A recent article published in the Journal of the American Academy of Dermatology reviewed adverse events regarding botulinum toxin type A (BTX-A) reported to the Food and Drug Administration between 1989 and 2003. Although postmarketing surveillance is a vital mechanism to ensure drug safety, the events reported in this paper must be considered in context to be appropriately interpreted. The majority of data was related to therapeutic rather than cosmetic use. The proportion of serious adverse events was 33-fold higher for therapeutic use and no deaths were reported after cosmetic use. The data were derived from a spontaneous reporting system and do not include assessments of causality between the BTX-A and purported adverse events. The report notes that over a third of these events were related to off-label use of BTX-A, a common practice in dermatology, yet no significant differences were reported in rates of adverse events between on-label and off-label use. The report reflects a favorable safety profile for cosmetic use of BTX-A, and if misinterpreted, could lead to unreasonable conclusions regarding a product considered to be highly safe and effective. PMID- 16310075 TI - Two cases of radiation-induced skin injuries occurring after radiofrequency catheter ablation therapy for atrial fibrillation. PMID- 16310076 TI - Penile nodule in a 54-year-old man: a case of a myointimoma. PMID- 16310077 TI - Drug-induced delusions of parasitosis during treatment of Parkinson's disease. PMID- 16310078 TI - Unusual clinical and dermoscopic features in two cases of pigmented basal cell carcinoma. PMID- 16310079 TI - Acute cyclosporine nephrotoxicity in a patient with oral pemphigus vulgaris and HIV infection on antiretroviral therapy. PMID- 16310080 TI - Extensive purpura in a patient with paroxysmal nocturnal hemoglobinuria. PMID- 16310081 TI - Ectopic nail with polydactyly. PMID- 16310082 TI - Rapid onset of CD8+ aggressive T-cell lymphoma during bexarotene therapy in a patient with Sezary syndrome. PMID- 16310083 TI - Lack of efficacy of topical latanoprost in the treatment of eyebrow alopecia areata. PMID- 16310084 TI - Keloids are not angiogenic lesions. PMID- 16310086 TI - Pityrosporum folliculitis during pregnancy is not pruritic folliculitis of pregnancy. PMID- 16310087 TI - Light's role in immunomodulation and medical therapy. PMID- 16310088 TI - Is oral pulsed prednisolone useful in alopecia areata? Critical appraisal of a randomized trial. PMID- 16310090 TI - Diet and acne--slim evidence, again. PMID- 16310091 TI - Dairy and acne--the iodine connection. PMID- 16310093 TI - Rosacea, acne rosacea, and actinic telangiectasia: in reply. PMID- 16310094 TI - Pyoderma faciale. PMID- 16310096 TI - Detrimental effects of high plasma urea nitrogen levels on viability of embryos from lactating dairy cows. AB - High plasma urea nitrogen (PUN) concentrations are associated with decreased fertility in lactating dairy cows. Our objective was to evaluate the quality of embryos flushed from superovulated lactating cows having moderate or high PUN concentrations. Subsequent embryo survival was determined after transfer to recipient heifers with either low or high PUN. Lactating Holstein dairy cows (n = 23; 50-120 days in milk) were randomly assigned to one of two diets designed to result in moderate or high PUN concentrations (15.5 +/- 0.7 and 24.4 +/- 1.0 mg/dl, respectively; P < 0.001) and were fed for 30 days before embryo flushing and recovery. Embryos (n = 94) were evaluated morphologically, frozen and subsequently transferred into synchronized virgin heifers that were fed one of two diets designed to result in either low or high PUN concentrations (7.7 +/- 0.9 and 25.2 +/- 1.5 mg/dl, respectively; P < 0.001; 2 x 2 factorial design). The number, quality and stage of development of recovered embryos were similar for cows with moderate or high PUN. Transfer of embryos from moderate PUN donor cows resulted in a higher pregnancy rate (35%; P < 0.02) than the transfer of embryos from high PUN donor cows (11%). Pregnancy rate was not affected by either recipient diet or the interaction of donor and recipient diets (P > 0.05). These results indicate that high PUN concentrations in lactating dairy cows decrease embryo viability through effects exerted on the oocyte or embryo before recovery from the uterus 7 days after insemination. PMID- 16310097 TI - Genetic and environmental factors influencing first service conception rate and late embryonic/foetal mortality in low fertility dairy herds. AB - The objective of this study was to identify factors affecting variation in conception rate to first artificial inseminations (AI) (CR: number of pregnant cows on D80-100/inseminated cows) and the incidence of embryonic/foetal loss (LEM) between 21 and 80 days of pregnancy (number of cows non-pregnant on D80 100/pregnant on D21) in 44 low fertility dairy herds of the west-central region of France. Reproductive status was assessed using progesterone milk concentration on D0 = Day of AI and D21-24, plasma PSPB concentration on D30-35, rectal palpation on D80-100 and observed return to oestrous. The final data set contained 1285 Prim'Holstein cows, 5.0% (64/1285) were inseminated in the luteal phase (progesterone > or = 3 ng/ml on D0), 61.3% (787/1285) were pregnant on D21 24 (progesterone < 3 ng/ml on D0 and > or = 5 ng/ml on D21-24), 15.4% lost their embryo/foetus between D21-24 and D80-100 (198/1285) and 45.8% (589/1285) were pregnant on D80-100. The incidence of late embryonic/foetal loss (LEM) was 25.2% (198/787). Multivariate logistic regression models including the random herd effect were used to analyse the relationship between AI centre, AI sire, cow's sire, parity, interval between calving and AI, milk production, milk protein content, body condition score (BCS) on D0, season of calving, season of AI, estimated genetic index on CR and LEM incidence. CR was significantly related to parity (p < 0.05), milk production after calving (p < 0.05) and estimated genetic value (p < 0.01). A significant difference in CR was observed for calving to AI interval > or = 70 days versus > or = 90 days, but the overall effect of the interval was not significant (p = 0.11). LEM incidence was affected by period of AI (p < 0.05), milk production (p < 0.05) and BCS (p < 0.05), but was not related to estimated genetic index. In conclusion, in these low fertility herds, the incidence of LEM was high and 25% of the cows lost their embryo after 21 days of pregnancy. LEM was affected by specific factors (season, BCS), which were not related to CR. The absence of a relationship between estimated genetic index and LEM in spite of its effect on CR indicates that estimated genetic merit has a greater effect on early embryonic loss or fertilisation failure than on later stages of embryo development. PMID- 16310098 TI - Deficiency of uridine monophosphate synthase (DUMPS) and X-chromosome deletion in fetal mummification in cattle. AB - Ten mummified fetuses were tested for the deficiency of uridine monophosphate synthase (DUMPS), which is known to contribute to the embryonic and fetal mortality in cattle. Genomic DNAs of the mummified fetuses were extracted from tissue samples collected from the mummies and were amplified by GenomiPhi DNA amplification kit. UMPS gene of the mummies was amplified by polymerase chain reaction (PCR) with DUMPS primers. Out of ten mummies examined, two fetuses were heterozygous (carriers) for DUMPS as indicated by the presences of three bands of 89, 53 and 36 bp. Estimated stage of gestation when the death occurred in the two mummies was 3.5 and 2.5 months, respectively. The other fetuses exhibited only two bands of 53 and 36 bp on the polyacrylamide gel indicated that they were normal. On the other hand, all the mummies were sexed using AMX/Y primers. Specific regions of Y and X chromosomes were amplified by PCR using AMX/Y. The expected 280 bp fragment in the female sample and the 280 and 217 bp in the male sample were observed. Nine mummies had a normal X and Y chromosome bands; however, the other mummified fetus exhibited only Y chromosome band, while the constitutive X chromosome fragment was missing. The estimated stage of gestation when the death occurred in this mummified fetus was 100 days. This might be the first report of DUMPS and X-chromosome deletion at the amelogenin gene in bovine mummified fetuses in Japan. PMID- 16310099 TI - Fertility-associated proteins in Nelore bull sperm membranes. AB - The present study was undertaken to evaluate the protein composition of the sperm membranes (SM) of Nelore bulls, assessing protein markers associated with bull fertility, and whether these markers can be used for predicting bull fertility. Samples were obtained of 20 Nelore bulls, with fertility ranked and divided into three groups (greater, normal and least). To rank the bull's fertility weighted classification was used (according to the number of pregnant cows, number of AI cows and number of herds, considering three different breeding seasons), using the PROC GENMOD as a statistical model, with 99% significance. A total of 7897 Nelore cows, randomly distributed among 28 different farms, were considered in the statistical analyses. The bulls were divided into three fertility groups (pregnancy rates): greater (%F > 80), normal (79 < %F > 71) and least (< 68%F) with 3, 13 and 4 bulls, respectively. Two-dimensional gel electrophoresis (2DE) of sperm membranes indicated in 27 spots (SM40, SM53, SM69, SM93, SM102, SM111, SM137, SM138, SM189, SM196, SM201, SM202, SM204, SM225, SM236, SM237, SM239, SM241, SM246, SM247, SM275, SM283, SM342, SM346, SM355, SM372, SM391) was prevalent in the higher fertility group, and just one spot (SM244) was prevalent in the lower fertility group. Spots SM244 and SM239 had their identification defined by PMF/MALDI-MS, as BSP-A3 and aSFP, respectively. Both these proteins showed a great potential for predicting bull's fertility. The amount of aSFP was 8.5 times greater in the sperm membrane protein profile of the higher fertility groups of Nelore bulls. Besides that, the BSP-A3 was 2.5 times greater in the lower fertility group. For the other spots potentially associated with fertility not yet identified, additional tests will be necessary, but it is clear that the 2D electrophoresis of the sperm membrane can be used for a new approach to predict Nelore bull fertility. PMID- 16310100 TI - LH and IGF-1 release during oestrus and early luteal phase in lactating and non lactating horse mares. AB - The aim of the present study was to determine effects of lactation on basal LH and IGF-1 concentrations and on the LH response to a GnRH-analogue at different stages of the oestrous cycle in mares. A total of 17 cyclic Haflinger mares were included in the study. Experiments were performed on lactating mares in first postpartum oestrus, the subsequent early luteal phase, and second postpartum oestrus. Non-lactating mares were used in oestrus and early luteal phase. Blood samples were taken for 1 h at 15 min intervals. Mares were then injected with the GnRH-analogue buserelin (GnRHa; 5 microg i.v.) and blood samples were drawn every 15 min for further 2 h. LH in all samples and basal IGF-1-concentrations were determined by RIA. In lactating mares, basal LH concentrations during the early luteal phase tended to be lower (p = 0.07) and the LH response to GnRHa, calculated as area under the curve, was significantly less pronounced compared to non-lactating mares (p < 0.01). As well in lactating mares, the basal LH concentration between first early luteal phase and second oestrus differed significantly (p < 0.05) and the net response to GnRHa was significantly lower between first oestrus as well as second oestrus and first early luteal phase (p < 0.05) but not between first and second oestrous postpartum. Within the group of non-lactating mares, the LH response to GnRHa was as well significantly lower during oestrus than during early luteal phase (p < 0.01). IGF-1 concentrations differed neither between groups nor stages of the cycle within groups. In conclusion, basal and GnRHa-stimulated LH release in lactating mares is lower than in non-lactating mares. This difference, however, occurs only in the early luteal phase. In lactating mares, concentrations of LH appear adequate to allow ovulation to occur. PMID- 16310101 TI - The involvement of protein kinases in signalling of opioid agonist FK 33-824 in porcine granulosa cells. AB - It is known that acute action of mu opioid receptor agonist, FK 33-824, results in an inhibition of oestradiol (E2) secretion by porcine granulosa cells from large follicles, but the opioid mode of action is unknown. In the present study, the involvement of two signal transduction pathways, phospholipase C/protein kinase C and adenylyl cyclase/protein kinase A, in mechanism of the opioid action was investigated. Treatment of pig granulosa cells with FK 33-824 at the dose 1 nM suppressed E2 secretion. Protein kinase C (PKC) inhibitors - staurosporine (1 100 nM), d-sphingosine (10-500 nM) and PKCi (100-2000 nM) - both alone and in combination with FK 33-824 reduced E2 release from the cells in relation to the control group. The inhibitory effect of the opioid on E2 output was also observed in PKC-deficient granulosa cells. PKC activator, PMA (10 and 100 nM) significantly attenuated the inhibitory effect of the opioid agonist. FK 33-824 also inhibited 3[H]phorbol 12,13 dibutyrate (3[H]PDBu) specific binding by granulosa cells. Adenylyl cyclase (AC) engagement in opioid signal transduction was assayed after 2-h and 4-h incubations of granulosa cells. During 2-h incubation, FK 33-824 at the dose 1 nM decreased cAMP secretion. Prolongation of the incubation up to 4 h caused disappearance of the opioid action. The addition of protein kinase A (PKC) inhibitor, PKAi (100-2000 nM), alone or together with FK 33-824, was followed by an inhibition of E2 secretion. FK 33-824 with the highest dose of PKAi (2000 nM) significantly inhibited E2 secretion by the cells in comparison to these agents tested separately. The opioid added in combination with PKA activator, 8BrcAMP (1000 microM), caused attenuation of stimulatory effect of 8BrcAMP. Collectively, these results suggest that acute action of mu opioid agonist on porcine granulosa cells leads to decrease of enzymatic activity of PKC and AC/PKA. It is not ruled out that other signal transduction pathways - not considered in this study - may also be engaged in the opioid mechanism of action in these cells. PMID- 16310102 TI - Ultrastructural localisation of calcium deposits in pig ovarian follicles. AB - Calcium intracellular signaling regulates many intracellular events including oocyte maturation. This signaling is strongly dependent on the influx of calcium ions from extracellular spaces and on the state of intracellular calcium stores. In this study, intracellular calcium deposits were detected in follicle-enclosed pig oocytes using the combined oxalate-pyroantimonate method. These deposits were observed in the nucleus, the mitochondria, the cytoplasm, and on the surface of lipid droplets. The amount of calcium deposits was expressed as a percentage of the area of the respective cellular compartment, which is covered with calcium deposits on ultrathin sections. The distribution of calcium deposits in oocytes changed during folliculogenesis. The amount of calcium deposits in nuclei (1.11% of the area of oocyte nuclei) and cytoplasm (1.02%) in oocytes from secondary and early antral follicles (0.90% nuclei; 0.99% cytoplasm) is significantly lower (P < 0.05) than the amount of calcium deposits in these compartments in oocytes from primary follicles (2.51% nuclei; 2.34% cytoplasm) or antral follicles with growing oocyte (2.91% nuclei; 2.21% cytoplasm). The amount of calcium deposits in mitochondria of oocytes from primary follicles (1.27%) or antral follicles with growing oocyte (1.14%) is significantly lower (P < 0.05) than in the nucleus (2.51% in oocytes from primary follicles; 2.91% in growing oocytes from antral follicles) or cytoplasm (2.34% in oocytes from primary follicles; 2.21% in growing oocytes from antral follicles). The amount of calcium deposits in the cytoplasm of fully-grown oocytes (1.46%) dropped to levels significantly lower (P < 0.05) than those observed in the oocyte nucleus (2.29%). On the basis of these data, we can conclude that the population of follicles on pig ovaries differs in the distribution and concentration of calcium deposits in oocytes, and these changes may be involved in the regulation of the meiotic competence of oocytes. PMID- 16310103 TI - Twenty-four hour rhythm of plasma prolactin in female rabbit pups. Correlation with hypothalamic and adenohypophysial dopamine, serotonin, gamma-aminobutyric acid and taurine content. AB - Lactation in the rabbit is a nocturnal activity, extremely short and regular, that can be a strong synchronizer for the development of circadian rhythmicity in the pups. In the present study, 24-h rhythmicity of plasma prolactin and median eminence and anterior pituitary content of dopamine (DA), serotonin (5HT), gamma aminobutyric acid (GABA) and taurine were examined in 11 days old female pups kept under 16 h light:8 h dark photoperiods (lights on at 08:00 h). Groups of six to seven female rabbit pups were killed by decapitation at six different time points throughout a 24-h cycle, starting at 09:00 h. Plasma prolactin levels changed significantly throughout the day, showing two peaks, one at first half of rest span (at 13:00 h) and another one at the beginning of the scotophase (at 01:00 h), just preceding doe visit. Median eminence DA content changed in a bimodal way as a function of time of day, displaying two maxima, at the beginning of the rest span and of the activity phase. Median eminence DA and plasma prolactin correlated significantly in an inverse way. Two maxima in median eminence 5HT levels were found, about 4 h in advance to the prolactin peaks. Circulating prolactin correlated inversely with median eminence 5HT content and directly with adenohypophysial 5HT content. Median eminence GABA content reached its maximum at the beginning of the scotophase and correlated significantly with plasma prolactin concentration. A positive correlation between plasma prolactin and adenohypophysial taurine content was observed. These results show that the circadian rhythmicity in prolactin secretory mechanisms in female rabbit pups develops during the early neonatal life. PMID- 16310104 TI - The intertubular compartment morphometry in capybaras (Hydrochoerus hydrochaeris) testis. AB - The aim of the present study was to characterize the intertubule element volume density, individual and total Leydig cells volume, Leydig cell number per testis and per gram of testis, and leydigosomatic index in adult capybaras. Eight capybaras from a commercial abattoir were utilized. The intertubular compartment volume density and the Leydig cells were 45.2 and 31.13%, respectively. The individual and total Leydig cell volumes were 8.51 and 2169.41 x 10(-12) mL, respectively. The Leydig cell number per testis was 3.8 billion and the Leydig cell number per gram of testis was 126 million. The leydigosomatic index was 0.037%. In conclusion, this study shows that capybaras have one of the greatest individual and total Leydig cell volume and Leydig cell volume density, and that the Leydig cell number per gram of testis is at least double the mean for mammals previously investigated in its order. PMID- 16310105 TI - Reconstruction as a source of artifact in non-gated single-shot diffusion weighted EPI. AB - A controversy has existed over the requirement to cardiac gate diffusion-weighted MRI acquisitions of the brain. Conventional wisdom suggests gating to be a necessary requirement to allow acquisition of accurate data, but recent applications find gating not necessary. The signal-to-noise and acquisition duration of these two approaches can be quite different; thus, this difference in methodology is important. This is particularly relevant when performing quantitative work such as diffusion tensor imaging. Here, the convention to gate is explained as being due to the historical use of low spatial resolution and more recently to the use of different reconstruction approaches. It is demonstrated that the Margosian reconstruction approach only yields high quality results when used in a gated fashion. Zero padding of the acquisition matrix provides an alternative reconstruction method that is not found to accentuate the artifacts that are due to pulsatile motion in the diffusion-weighted acquisition and thus do not require a gated acquisition. The relative merits of each reconstruction approach are discussed, including estimates of the relative signal to-noise ratio and resolution benefits. It is concluded that both gated methods and non-gated methods can each provide high quality results with appropriate reconstruction methods. PMID- 16310106 TI - Mapping of the cerebral response to acetazolamide using graded asymmetric spin echo EPI. AB - Cerebral vascular reactivity in different regions of the rat brain was quantitatively characterized by spatial and temporal measurements of blood oxygenation level-dependent (BOLD)-fMRI signals following intravenous administration of the carbonic anhydrase inhibitor acetazolamide: this causes cerebral vasodilatation through a cerebral extracellular acidosis that spares neuronal metabolism and vascular smooth muscle function, thus separating vascular and cerebral metabolic events. An asymmetric spin echo-echo planar imaging (ASE EPI) pulse sequence sensitised images selectively to oxygenation changes in the microvasculature; use of a surface coil receiver enhanced image signal-to-noise ratios (SNRs). Image SNRs and hardware integrity were verified by incorporating quality assurance procedures; cardiorespiratory stability in the physiological preparations were monitored and maintained through the duration of the experiments. These conditions made it possible to apply BOLD contrast fMRI to map regional changes in cerebral perfusion in response to acetazolamide administration. Thus, fMRI findings demonstrated cerebral responses to acetazolamide that directly paralleled the known physiological actions of acetazolamide and whose time courses were similar through all regions of interest, consistent with acetazolamide's initial distribution in brain plasma, where it affects cerebral haemodynamics by acting at cerebral capillary endothelial cells. However, marked variations in the magnitude of the responses suggested relative perfusion deficits in the hippocampus and white matter regions correlating well with their relatively low vascularity and the known vulnerability of the hippocampus to ischaemic damage. PMID- 16310107 TI - Nonlinear responses of cerebral blood volume, blood flow and blood oxygenation signals during visual stimulation. AB - Hemodynamic-based functional magnetic resonance imaging (fMRI) techniques provide a great utility for noninvasive functional brain mapping. However, because the hemodynamic signals reflect underlying neural activity indirectly, characterization of these signals following brain activation is essential for experimental design and data interpretation. In this report, the linear (or nonlinear) responses to neuronal activation of three hemodynamic parameters based primarily on changes of cerebral blood volume, blood flow and blood oxygenation were investigated by testing these hemodynamic responses' additivity property. Using a recently developed fMRI technique that acquires vascular space occupancy (VASO), arterial spin labeling (ASL) perfusion and blood oxygenation level dependent (BOLD) signals simultaneously, the additivity property of the three hemodynamic responses in human visual cortex was assessed using various visual stimulus durations. Experiments on healthy volunteers showed that all three hemodynamic-weighted signals responded nonlinearly to stimulus durations less than 4 s, with the degree of nonlinearity becoming more severe as the stimulus duration decreased. Vascular space occupancy and ASL perfusion signals showed similar nonlinearity properties, whereas the BOLD signal was the most nonlinear. These data suggest that caution should be taken in the interpretation of hemodynamic-based signals in fMRI. PMID- 16310108 TI - Age-related cerebral white matter changes and pulse-wave encephalopathy: observations with three-dimensional MRI. AB - Our purpose was to investigate leukoaraiosis (LA) using three-dimensional MR imaging combined with advanced image-processing technology to attempt to group signal abnormalities according to their etiology. Coronal T2-weighted fast fluid attenuated inversion-recovery (FLAIR) sequences and three-dimensional T1-weighted fast spoiled gradient recalled echo sequences were used to examine cerebral white matter changes in 75 elderly people with memory complaint but no dementia. They were otherwise healthy, community-dwelling subjects. Three subtypes of LA were defined on the basis of their shape, geography and extent: the so-called subependymal/subpial LA, perivascular LA and "bands" along long white matter tracts. Subependymal changes were directly contiguous with ventricular spaces. They showed features of "water hammer" lesions with ventricular systematisation and a more frequent location around the frontal horns than around the bodies (P=.0008). The use of cerebrospinal fluid (CSF) contiguity criterion allowed a classification of splenial changes in the subpial group. Conversely, posterior periventricular lesions in the centrum ovale as well as irregular and extensive periventricular lesions were not directly contiguous with CSF spaces. The so called perivascular changes showed features of small-vessel-associated disease; they surrounded linear CSF-like signals that followed the direction of perforating vessels. Distribution of these perivascular changes appeared heterogeneous (P ranging from .04 to 5.10(-16)). These findings suggest that subependymal/subpial LA and subcortical LA may be separate manifestations of a single underlying pulse-wave encephalopathy. PMID- 16310109 TI - Magnetic resonance cholangiopancreatography using a free-breathing T2-weighted turbo spin-echo sequence with navigator-triggered prospective acquisition correction. AB - PURPOSE: The objective of this study was to evaluate the image quality of a respiratory-triggered T2-weighted (T2w) turbo spin-echo (TSE) sequence for magnetic resonance cholangiopancreatography (MRCP) using a new method for respiratory triggering by tracking the motion of the right diaphragm [prospective acquisition correction (PACE) technique]. MATERIALS AND METHODS: Fifty consecutive patients underwent MRCP imaging applying breath-hold half-Fourier single-shot TSE sequences and the respiratory-triggered T2w TSE sequence. Qualitative evaluation grading the depiction of eight segments of the pancreaticobiliary tree and the frequency of artifacts was performed. Quantitative evaluation included calculation of the relative contrast (RC) between fluid-filled ductal structures and organ parenchyma at four segments. RESULTS: A significantly higher (P<.01) RC was measured for the respiratory triggered T2w TSE sequence [maximum intensity projection (MIP)] for all of the four investigated segments (one of four segments for the MIP) of the pancreaticobiliary tree, as well as a significant (P<.01) improvement of visualization of all ductal segments compared with the breath-hold sequences. The frequency of artifacts was significantly lower (P<.01) compared with the breath hold sequences. CONCLUSION: Respiratory-triggered MRCP using a T2w TSE sequence with PACE significantly improves image quality and may be included into the routine MRCP sequence protocol. PMID- 16310110 TI - Susceptibility-matched envelope for the correction of EPI artifacts. AB - Fast gradient echo sequences, such as echo planer imaging (EPI) and spiral imaging, are vulnerable to artifacts resulting from B(0) inhomogeneities. A major contribution to these artifacts is the susceptibility variation across the head, which is most severe in regions adjacent to air-tissue interfaces, such as the mouth, nasal sinuses, ears and the cortex. Susceptibility artifacts can cause geometrical distortions in the image as well as loss of signal due to T(2)* dephasing. The extent of these artifacts increases with the main field, thus compromising the signal-to-noise ratio (SNR) benefit gained in higher fields. In the current work, inhomogeneity caused by susceptibility variations at the external boundary of the human body has been corrected by surrounding the organs with a liquid without hydrogen atoms and whose susceptibility is similar to that of the imaged organ. EPI experiments were conducted on head-sized phantom, human brain, hand and legs. This method causes minimal patient inconvenience and no interference with any function of the scanner, thus yielding a simple and efficient solution for the correction of B(0) variation. PMID- 16310111 TI - Implications of the Rician distribution for fMRI generalized likelihood ratio tests. AB - In functional magnetic resonance imaging (fMRI), the general linear model test (GLMT) is widely used for brain activation detection. However, the GLMT relies on the assumption that the noise corrupting the data is Gaussian distributed. Because the majority of fMRI studies employ magnitude image reconstructions, which are Rician distributed, this assumption is invalid and has significant consequences in case the signal-to-noise ratio (SNR) is low. In this study, we show that the GLMT should not be used at low SNR. Furthermore, we propose a generalized likelihood ratio test for magnitude MR data that has the same performance compared to the GLMT for high SNR, but performs significantly better than the GLMT for low SNR. PMID- 16310112 TI - Changes in intratumor heterogeneity in blood perfusion in intradermal human melanoma xenografts during tumor growth assessed by DCE-MRI. AB - The purpose of this study was to use dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) to search for systematic intratumor heterogeneity in blood perfusion in human melanoma xenografts growing intradermally in BALB/c-nu/nu mice. Six xenografted tumors of an amelanotic human melanoma line (A-07) were included in the study. DCE-MRI was performed daily for 5 days by using spoiled gradient recalled sequences. Tumor images of E.F (E is initial extraction fraction and F is perfusion) were produced by subjecting DCE-MRI data to Kety analysis. E.F was used as a measure of tumor blood perfusion, since comparative studies have shown that E.F is closely related to blood perfusion in A-07 tumors. The E.F images indicated that the intratumor heterogeneity in blood perfusion was similar in all investigated tumors. The blood perfusion was low in the center of the tumors and increased toward the tumor periphery in the dorsal and ventral direction by a factor of 3-4, but not in the lateral and medial direction. The magnitude of the heterogeneity increased by a factor of approximately 2 during tumor growth. In conclusion, intradermal human melanoma xenografts show significant systematic intratumor heterogeneity in blood perfusion. PMID- 16310113 TI - Biodistribution and pharmacokinetics of the (19)F-labeled radiosensitizer 3 aminobenzamide: assessment by (19)F MR imaging. AB - 3-Aminobenzamide (3-ABA) is a potent radiosensitizer that inhibits the repair of DNA strand breaks. The aim of this study was to monitor the biodistribution and pharmacokinetics of a fluorinated 3-ABA derivative in tumor-bearing rats by magnetic resonance imaging (MRI). To this end, 3-ABA was labeled with fluorine-19 by trifluoroethylation [3-amino-N-2,2,2-trifluoroethylbenzamide (3-ABA-TFE)], which only slightly increased the cytotoxicity of the compound as demonstrated by colony-forming assays. After intraperitoneal injection of 400 mg/kg BW 3-ABA-TFE to nine Copenhagen rats with Dunning prostate adenocarcinoma, (19)F MR images were acquired at a whole-body MR system with a spatial sampling of 10 x 10 x 15 mm(3). While 3-ABA-TFE was observed in all major organs and the muscular system, only a small and heterogeneous signal could be detected in the adenocarcinoma. Serial MR measurements yielded maximum tissue signals about 2 days after 3-ABA TFE administration. At this time point, the mean muscle-to-liver and tumor-to liver signal ratio was 0.31+/-0.07 and 0.11+/-0.04, respectively. Application of the (19)F MRI strategy makes it possible to measure the biodistribution and pharmacokinetics of 3-ABA-TFE in individual animals in a longitudinal manner. The results obtained for the prostate adenocarcinoma indicate that delivery of 3-ABA TFE to solid tumors may be seriously hampered by tumor-specific factors and that the intratumoral uptake of the substance may be lower than in normal tissues. Therefore, the development of effective carrier systems is mandatory to improve tumor-selective delivery. PMID- 16310114 TI - Effects of the tsunami on waste management in Sri Lanka. PMID- 16310115 TI - On the effect of demographic characteristics on the formulation of solid waste charging policy. AB - This paper presents the results from an analysis of municipal charges in Greece, as the fees paid by municipal waste producers to the municipal authorities are included in these charges and constitute a major part of them. The aim is to determine whether and how some of the demographic characteristics of a municipality influence the formulation of municipal waste charges and specifically the level of the charge. The study included 339 Hellenic municipalities, which comprised approximately 62% of the overall population in the country. PMID- 16310116 TI - Leachate migration from spent mushroom substrate through intact and repacked subsurface soil columns. AB - Field weathering of spent mushroom substrate (SMS) produces soluble compost leachate that percolates into underlying soils and may adversely impact groundwater. Laboratory experiments were conducted to investigate movement and retention of SMS leachate solutes in subsurface soil columns. Spent mushroom substrate leachate with high concentrations of dissolved organic matter (DOM) and inorganic salts was passively loaded to intact and repacked columns of Bt1 soil (fine-loamy, mixed, semiactive, mesic Typic Hapludults) and effluents were monitored for changes in chemical composition. Transport of SMS leachate in undisturbed soil cores was mainly via preferential flow, whereas matrix flow was predominant in repacked soil columns. Leachate DOM and phosphate were sorbed by soil minerals while Cl-, SO4(2-), Na+ and NH4+ were eluted. Leachate K+ displaced exchangeable native cations and was retained. Biodegradation of leachate DOM resulted in reduction and elution of soil Mn and Fe, especially in repacked columns. Persistent anoxia also inhibited nitrification. Precipitation of gypsum and CaCO3 blocked preferential flow channels, and movement of SMS leachate was subsequently reduced. The results demonstrate that SMS leachate migrates via rapid preferential flow initially, followed by matrix flow at a lower rate. Leachate solutes may transport to depth in soil profiles through preferential channels. To protect water resources, weathering of deep SMS piles should be conducted on compact surfaces or in fields with a condensed soil layer (no structural cracks) above the groundwater table, and measures controlling leachate runoff be imposed. PMID- 16310117 TI - Anaerobic co-digestion of coffee waste and sewage sludge. AB - The feasibility of the anaerobic co-digestion of coffee solid waste and sewage sludge was assessed. Five different solid wastes with different chemical properties were studied in mesophilic batch assays, providing basic data on the methane production, reduction of total and volatile solids and hydrolysis rate constant. Most of the wastes had a methane yield of 0.24-0.28 m3 CH4(STP)/kg VS(initial) and 76-89% of the theoretical methane yield was achieved. Reduction of 50-73% in total solids and 75-80% in volatile solids were obtained and the hydrolysis rate constants were in the range of 0.035-0.063 d(-1). One of the solid wastes, composed of 100% barley, achieved a methane yield of 0.02 m3 CH4(STP)/kg VS(initial), reductions of 31% in total solids, 40% in volatile solids and achieved only 11% of the theoretical methane yield. However, this waste presented the highest hydrolysis rate constant. Considering all the wastes, an inverse linear correlation was obtained between methane yield and the hydrolysis rate constant, suggesting that hydrolysis was not the limiting factor in the anaerobic biodegradability of this type of waste. PMID- 16310118 TI - Method to recover and reuse chromated copper arsenate wood preservative from spent treated wood. AB - The volume of chromated copper arsenate (CCA) treated wood products coming out of service is expected to increase dramatically during the next decade. There is a need for an alternative waste management approach to landfilling. This paper investigates the variables affecting extraction of CCA components from wood particles and the potential to oxidize and reuse the recovered chemicals. Most of the CCA components could be extracted by 10% H2O2 at 50 degrees C in 6 h with an average extraction efficiency of 95% for Cr, 94% for Cu and 98% for As. The extract containing Cr(III), Cu(II) and As(V) could be oxidized in several stages by aqueous 2.5% w/w H2O2 in less than 2 h to a condition where it was compatible with CCA treating solutions and could be reused for treating new wood. When the recovered extract was mixed with fresh CCA solution in different ratios, the mixed CCA-C solutions had similar solution stability as freshly prepared CCA-C solution and treated wood had similar leaching properties as wood treated with fresh solution. PMID- 16310119 TI - Is the medication bottle for pediatric and adolescent depression half-full or half-empty? PMID- 16310120 TI - Depression treatment during outpatient visits by U.S. children and adolescents. AB - PURPOSE: Depression affects approximately 2-8% of all children and adolescents, and treatment of depression in children and adolescents has been the center of recent serious debates. We examined national trends in depression visits and treatment among outpatients aged 7 to 17 years. METHODS: We analyzed visit-based data between 1995 and 2002 in two national ambulatory care surveys. RESULTS: The number of visits by children and adolescents during which depression was reported more than doubled from 1995-1996 (1.44 million) to 2001-2002 (3.22 million). The proportion of these visits during which antidepressants were prescribed rose slightly from 47% in 1995-1996 to 52% in 2001-2002, whereas the proportion during which psychotherapy or mental health counseling was provided declined from 83% to 68%. Selective serotonin reuptake inhibitors (SSRI) represented 76% of all antidepressants prescribed in 1995-1996 and 81% in 2001-2002. In absolute terms, SSRIs were reported in 1.35 million visits in 2001-2002, reflecting a 2.6-fold increase from 1995-1996. Fluoxetine was prescribed in 207,914 visits in 1995-1996 and increased 100% to 415,580 visits in 2001-2002. The use of sertraline increased by 62% to 345,576 visits and paroxetine by 269% to 279,275 visits. CONCLUSIONS: We observed a declining trend in the provision of psychotherapy/mental health counseling during outpatient visits by children and adolescents diagnosed with depression. Although the likelihood of receiving antidepressants remained essentially unchanged, the number of children and adolescents whose visits involved prescription of antidepressants, particularly SSRIs, has increased markedly through 2002. Although fluoxetine remained the most commonly prescribed, other SSRIs were increasingly prescribed through 2002. These trends raise concerns regarding the widespread off-label use of antidepressants lacking reliable evidence of safety and efficacy for use in children and adolescents. PMID- 16310121 TI - Suicidality and correlates among rural adolescents of China. AB - PURPOSE: Despite high rates of suicide in rural youths of China, little is known about the epidemiological characteristics of suicidal behaviors among this group of population. This study examined the prevalence and correlates of suicidality among Chinese adolescents. METHODS: An epidemiological survey was conducted in a sample of 1,362 adolescents from 5 high schools in a prefecture of Shandong, China. We obtained data using a structured questionnaire asking about suicidal behaviors, life stress, locus of control, depression, aggression, and individual and family characteristics. Data were analyzed using logistic regression models. RESULTS: Overall, 19% of the sample reported having suicidal ideation, and 7% reported having made a suicide attempt during the past 6 months. The prevalence rates of suicidal behaviors significantly increased over age. Female adolescents were more likely to report suicidal ideation than males, and older females (ages 16-18) were more likely to report suicide attempts than older males. Multivariate logistic regression models showed that female gender, older age, boarding in school, life stress, depression, and external locus of control were significantly associated with increased risk for suicidal ideation, while older age, life stress, external locus of control, poor academic performance, depression, and aggression were related to suicide attempts. CONCLUSIONS: Suicidal behaviors are common problems in rural adolescents of China. Female adolescents are particularly prone to report suicidal ideation and suicide attempts as they age. Multiple psychosocial factors are associated with suicidal behaviors, and psychopathology and life stress may play major roles in suicidal behaviors among rural adolescents of China. PMID- 16310122 TI - Stages of change in adolescents: an examination of self-efficacy, decisional balance, and reasons for relapse. AB - PURPOSE: The purpose of this study was to test selected constructs of the transtheoretical model (TTM) of behavior change in relation to exercise behavior with an adolescent sample. A further purpose was to examine reasons why adolescents stop physical activity and to relate these to stages of exercise change. METHODS: Participants were 15 to 17 year old students at a private high school (N = 327). Participants completed a questionnaire consisting of an exercise measure, measures of the TTM variables, and an open-ended question that asked if they used to exercise in the past but currently did not, why they had stopped. The TTM data were analyzed using ANOVA F-tests with post-hoc Scheffe tests. To evaluate the ability of the TTM variables to discriminate between stages of exercise behavior, standard discriminant function analyses were performed. Open-ended answers to the relapse question were themed according to a model which categorizes barriers preventing youth's participation in recreational activities as infrastructural, superstructural, and procedural. The relationship of perceived barriers to stage of behavior change was assessed using cross tabulation and chi-square analyses. RESULTS: Moderate support for the TTM constructs were found, with the strongest discriminator between stages being strenuous exercise, and self-efficacy being the most supported construct. Infrastructural, superstructural, and procedural barriers were all evident. CONCLUSION: Over 30% of previously active adolescents in this study reported barriers precluding extracurricular physical activity. Interventions should target the most often cited reasons for relapse: time and limits relating to the nature of the adolescent self. PMID- 16310123 TI - Psychological symptoms and nonfatal unintentional injuries among Chinese adolescents: a prospective study. AB - PURPOSE: To describe patterns of nonfatal unintentional injuries and investigate the association of psychological symptoms with injury risk among adolescents in Guangxi, China. METHODS: Adolescents were selected from middle and high school students in Baise City, Guangxi, China. Psychological symptoms at baseline were measured using SCL-90-R and nonfatal unintentional injuries that occurred among participants were recorded using a standardized injury surveillance questionnaire. Annual injury rates per 100 adolescents by adolescents' demographic and family characteristics were calculated. Student's t-test and the Chi-square test were used to test differences in mean raw scores of SCL-90-R subscales and injury rates. Relative risks and odds ratios of injury with 95% confidence intervals were calculated in univariate analyses and multivariate logistic models. RESULTS: A total of 1474 middle and high school students finished the study. Of them, 442 reported nonfatal unintentional injuries during the study period, resulting in an annual injury rate of 30.3 per 100 adolescents. Boys had a significantly higher injury rate than girls (32.6% vs. 27.4%). Injury rate decreased as the adolescents' age increased. Compared with adolescents who were not injured, the injured adolescents had higher average raw scores of all subscales of SCL-90-R, with an exception of the subscale for hostility. All SCL 90-R subscales factors, except those for hostility and phobia, were associated with injury risk. After confounding effects of age, gender, and ethnicity were controlled using logistic regression models, SCL-90-R subscales for somatization (OR = 2.00, 95% CI 1.52-2.63), obsessive-compulsiveness (OR = 2.10, 95% CI 1.71 2.58), interpersonal-sensitivity (OR = 1.66, 95% CI 1.34-2.06), depression (OR = 2.00, 95% CI 1.59-2.51), anxiety (OR = 2.08, 95% CI 1.62-2.66), and psychoticism (OR = 1.60, 95% CI 1.26-2.03) were significantly associated with an elevated injury risk. CONCLUSION: Psychological symptoms are associated with an elevated risk of nonfatal unintentional injury among middle and high school students. Mental health services, such as psychological counseling, may help reduce the risk of nonfatal unintentional injuries among Chinese adolescents. PMID- 16310124 TI - Factors that influence adolescent reports of counseling by physicians on risky behavior. AB - PURPOSE: To determine factors that affect whether low-income adolescents report that their doctor talked with them about risky behavior. METHODS: Random digit dial survey of low-income adolescents in New York City asking about depression, smoking, alcohol use, and sexual activity and the screening and counseling they received on these risk factors and risks during health visits. RESULTS: Prevalence of counseling by physicians was low, according to adolescent reports, ranging from 17% of adolescents counseled about depression to 52% about sexually transmitted diseases. Older adolescents were more likely than younger to receive counseling about all topics. In bivariate and multivariate models, having the risk factor was strongly associated with physicians counseling for depression (adjusted [adj.] OR = 4.42; p < 0.001); for sexual activity and counseling about condom use (adj. OR = 4.06; p < 0.01), and birth control (adj. OR = 2.76; p < 0.03). Still, many adolescents at risk had not received counseling. Many adolescents have not had a private and confidential visit with their provider. Having a private and confidential visit was also associated with receipt of counseling. CONCLUSIONS: Adolescents are not receiving sufficient counseling about risks and risky behavior, according to their own reports. There is need to improve delivery of counseling and ensure that private and confidential visits are provided to youth. PMID- 16310125 TI - Depression and role impairment among adolescents in primary care clinics. AB - PURPOSE: To evaluate the association between depression and role impairment in a primary care sample, with and without controlling for the effects of general medical conditions. METHODS: Cross-sectional survey of consecutive primary care patients, ages 13-21 years (n = 3471), drawn from six sites including public health, managed care, and academic health center clinics. We assessed probable depressive disorder, depressive symptoms, and common medical problems using youth self-report on a brief screening questionnaire. Main outcome measures were two indicators of role impairment: (a) decrement in productivity/role activity, defined as not in school or working full time; and (b) low educational attainment, defined as more than 2 years behind in school or > or = 20 years of age and failed to complete high school. RESULTS: Adolescents screening positive for probable depressive disorder had elevated rates of productivity/role activity decrements (19% vs. 13%; OR 1.69; 95% confidence interval [CI] 1.39-2.06; p < 0.001) and low educational attainment (20% vs. 15%; OR 1.47; 95% CI 1.21-1.78; p < 0.001). Probable depressive disorder made a unique contribution to the prediction of these impairment indicators after adjusting for the effect of having a general medical condition; controlling for depression, the presence of a general medical condition did not contribute to role impairment. CONCLUSIONS: Adolescent primary care patients screening positive for depression are at increased risk for impairment in school/work productivity and educational attainment. These findings emphasize the importance of primary care clinicians' attention to depression and role limitations. PMID- 16310126 TI - Social disadvantage and adolescent stress. AB - PURPOSE: Low socioeconomic status (SES) and minority race/ethnicity are both associated with chronic stress and co-vary in American society. As such, these factors are often used synonymously, without clear theoretical conceptualization of their roles in the development of stress-related health disparities. This study theorized that race/ethnicity and SES reflect social disadvantage, which is the underlying factor in the development of stress-related illness, and examined how social disadvantage, defined in terms of both race/ethnicity and SES, influences adolescents' stress. METHODS: This is a cross-sectional school-based study of 1209 non-Hispanic black and white 7th-12th graders from a single Midwestern metropolitan public school district. Each student completed a questionnaire and a parent provided SES information. Race/ethnicity was obtained from school records. Linear regression analyses determined the influence of race/ethnicity and SES to stress. Race/ethnicity and presence or absence of at least one parent who graduated from college were used to define four subgroups for within-group analyses. RESULTS: Stress was higher among black students, those from lower SES families, and those with lower perceived SES. In subgroup analyses, neither race nor SES maintained their independent associations with stress among socially disadvantaged groups. Black race was not associated with stress among those without a college-educated parent, and parent education did not influence stress among black students. In contrast, among more socially advantaged groups, both SES and race explained variation in adolescents' stress. CONCLUSIONS: Social disadvantage is associated with increased stress, regardless of whether disadvantage is defined in terms of race or SES. This suggests that race and SES measure adversity in the social environment, and therefore, serve as risk markers, rather than risk factors. Future research should focus on the experience of adversity, which is reflected by these social characteristics, and the processes by which it operates. PMID- 16310127 TI - Externalizing behaviors as predictors of substance initiation trajectories among rural adolescents. AB - PURPOSE: The purpose of the current study was to investigate the influence of externalizing behaviors on substance initiation trajectories among rural adolescents over a 42-month period. METHODS: Data were obtained from 198 rural adolescents (105 boys, 93 girls) who were participating in a longitudinal study. At the baseline assessment, subjects were on average 12.3 years of age. RESULTS: Controlling for gender, higher baseline levels of externalizing were associated with a greater number of substances initiated over time. The initiation trajectory was curvilinear. Girls, compared with boys, reported a lower number of substances initiated at baseline, a greater linear growth trajectory, and a deceleration of growth over time. CONCLUSIONS: The influence of adolescent externalizing behaviors on baseline levels and growth trajectories of substance initiation and the utility of latent growth curve modeling in the study of longitudinal change are discussed. PMID- 16310128 TI - Pediatricians' intention to administer human papillomavirus vaccine: the role of practice characteristics, knowledge, and attitudes. AB - PURPOSE: The objective of this study was to examine pediatrician characteristics and attitudes associated with intention to recommend two hypothetical human papillomavirus (HPV) vaccines. METHODS: A survey instrument mailed to a random sample of 1000 pediatricians assessed provider characteristics, HPV knowledge, and attitudes about HPV vaccination. Intention to administer each of two HPV vaccines types (a cervical cancer/genital wart vaccine and a cervical cancer vaccine) to girls and boys of three different ages (11, 14, and 17 years) was assessed. Linear mixed modeling for repeated measures and multivariable linear regression models were performed to identify variables associated with intention to recommend vaccination. RESULTS: The mean age of participants (n = 513) was 42 years and 57% were female. Participants were more likely to recommend vaccination to girls vs. boys and older vs. younger children, and were more likely to recommend a cervical cancer/genital wart vaccine than a cervical cancer vaccine (p < .0001). Variables independently associated with intention to recommend a cervical cancer/genital wart vaccine were: higher estimate of the percentage of sexually active adolescents in one's practice (beta .084, p = .002), number of young adolescents seen weekly (beta 1.300, p = .015), higher HPV knowledge (beta 1.079, p = .015), likelihood of following the recommendations of important individuals and organizations regarding immunization (beta .834, p = .001), and fewer perceived barriers to immunization (beta -.203, p = .001). CONCLUSIONS: Vaccination initiatives directed toward pediatricians that focus on modifiable predictors of intention to vaccinate, such as HPV knowledge and attitudes about vaccination, may facilitate adherence to emerging national immunization guidelines. PMID- 16310129 TI - The impact of a school entry law on adolescent immunization rates. AB - PURPOSE: Middle school entry laws increase coverage with recommended vaccines, but their effect on vaccines that are not required is unknown. We compared vaccination coverage for hepatitis B, tetanus and diphtheria (Td), and measles, mumps and rubella (MMR) in areas of states with discordant middle school, hepatitis B school entry laws, and evaluated the relationship between demographic characteristics and adolescent immunization rates. METHODS: Retrospective design with purposive school sampling, using location of residence to determine study group. In each school, immunization records from a random sample of up to 75 students in ninth grade (affected by a new hepatitis B law) and 12th grade (not affected by the law) from 11 schools in two areas discordant for the law were analyzed. All areas had long standing two-dose MMR and Td requirements. RESULTS: Ninth graders in schools with the law had hepatitis B rates higher (72.8%) than those without the law (18.6%) (U = 2.0, p < .01). There were no significant differences between grades or schools for MMR and Td. However, even in the presence of the law, rates were significantly lower in schools with lower socioeconomic indicators. CONCLUSIONS: Middle school immunization laws are effective at raising adolescent hepatitis B, but in this study there wasn't enough power to discern the effect on rates for other vaccines or the influence of demographic variables on rates. Results suggested that laws did not appear to completely overcome disparities. For school mandates to be more effective, additional efforts, presumably on enforcement, especially in areas with lower socioeconomic indicators, are needed. PMID- 16310130 TI - Peer activity in the evenings and participation in aggressive and problem behaviors. AB - PURPOSE: Low adult supervision during the after-school hours has been associated with numerous problem behaviors among youth. We examined the extent to which this relationship pertains to the evening hours and aggressive behaviors. METHODS: Cross-sectional self-report data were obtained from a nationally representative sample of 14,818 youth in grades 6-10 in the 2001-2002 Health Behaviors of School aged Children Survey. The relationship between spending evenings out with friends and involvement in problem behaviors was examined using logistic regression analyses. RESULTS: One-fifth of U.S. youth surveyed reportedly spent five or more evenings out with friends each week. After adjusting for grade, race/ethnicity, parental education, parental involvement, and perception of neighborhood safety, boys and girls who reported spending five or more evenings out were 4.3 and 3.1 times, respectively, than boys and girls who spent less than two evenings out, more likely to be involved in four or more physical fights in the past year; 3.0 and 4.0 times, respectively, more likely to have bullied another at least once a week at school; 2.7 and 4.9 times, respectively, more likely to have carried a weapon 6 or more days in the past month; 3.8 and 4.8 times, respectively, more likely to consume alcohol at least once a month; and 3.3 and 7.2 times, respectively, more likely to have smoked every day. CONCLUSIONS: Although the majority of youth who spend most evenings out do not frequently participate in problem behaviors (69.7%), their consistently increased risk for substance use and aggressive behaviors warrants attention. Further examination of specific evening activities, extracurricular involvement, neighborhood context, adult supervision, and parental monitoring is required. PMID- 16310131 TI - Aggressive behaviors in early adolescence and subsequent suicidality among urban youths. AB - PURPOSE: To examine the association of aggressive behaviors during early adolescence with subsequent suicidality among inner-city males and females. METHODS: Seven hundred sixty nine African-American and Latino males and females were surveyed about their involvement in aggressive behaviors in 8th grade and again during 11th grade, when suicidality was assessed. Logistic regression was used to examine the influence of middle-school aggression on subsequent suicidality, controlling for demographic and social characteristics and 8th grade depressive symptoms and substance use. At 8th grade, 43% of males and 32% of females reported a recent fight, 33% of males and 19% of females carried a knife, 8% of males and 7% of females used a knife to cut or stab someone, and 15% of males and 4% of females carried a gun. At 11th grade, 24% of males and 29% of females scored high on suicidality, indicating suicidal thoughts, plans, or attempt. RESULTS: Females scoring high on aggression were significantly more likely to report suicidality at 11th grade, controlling for demographic and social factors, early depressive symptoms, and substance use. For males, high depressive symptoms and substance use at 8th grade were related to suicidality, but not earlier aggressive behavior. CONCLUSION: Urban girls who engage in physical aggression in middle school are at risk for subsequent inner-directed violence, including suicidal thoughts and behaviors. To address adolescents' mental health needs, attention must be paid to connections between externally and internally directed forms of violence over time, and whether warning signs of suicidality may differ by gender and require different intervention. PMID- 16310132 TI - Immunogenicity and reactogenicity of a reduced-antigen-content diphtheria-tetanus acellular pertussis vaccine in healthy Taiwanese children and adolescents. AB - PURPOSE: Disease caused by Bordetella pertussis is increasingly being identified among older children and adults in immunized populations, indicating a waning of the vaccine-induced immunity. These findings suggest the need for booster immunization of older children and adults. Modern acellular reduced-antigen content vaccines have been developed, which can be given as a booster in individuals more than 4 years of age. This study was to assess the immunogenicity and reactogenicity of Boostrix, GlaxoSmithKline Biologicals' reduced-antigen content diphtheria-tetanus acellular pertussis (dTpa) vaccine, when administered as a booster in healthy subjects previously primed with DTP vaccine. METHODS: Healthy Taiwanese children and adolescents aged 6-8 years and 15-20 years, previously primed with DTP vaccine, were enrolled. All received one dose of Boostrix. Two blood samples were taken from each of them, one before vaccination and one at 1 month after vaccination. Serum antibodies to diphtheria and tetanus toxoids and immunoglobulin G (IgG) antibodies against the pertussis components PT, FHA and PRN were measured by enzyme-linked immunosorbent assay (ELISA) technique. Adverse reactions following vaccination were recorded. RESULTS: A total of 180 subjects were recruited. The vaccine response rates to the pertussis antigens ranged between 89.0-100%. There were no serious adverse events reported during the study period. CONCLUSIONS: The results of this study suggest that Boostrix may be safely and effectively administered as a booster dose to children previously primed with DTP vaccine. PMID- 16310133 TI - Premenstrual dysphoric disorder in adolescents: case reports of treatment with fluoxetine and review of the literature. AB - Premenstrual dysphoric disorder (PMDD) is a periodic, recurrent, debilitating condition with severe psychological or affective symptoms during the late luteal phase. PMDD often begins during adolescence. Dysregulation of the serotonergic system has been proposed recently as its cause and fluoxetine has been recommended as an appropriate treatment. We report 3 adolescents with PMDD who were treated for 2 years with fluoxetine, resulting in complete symptom resolution, and review the clinical trials supporting its use. Case reports of successfully treated teenagers are an addition to the accumulated evidence of the efficacy of fluoxetine for treatment of PMDD in adult women. Together they may provide some justification for the compassionate use of fluoxetine for adolescent girls who are being disabled by PMDD. However, more research is called for: a randomized placebo-controlled trial in adolescents is warranted. PMID- 16310134 TI - Self-reported symptoms of depression in late adolescence to early adulthood: a comparison of African-American and Caucasian females. AB - The prevalence of depressive symptoms from adolescence through young adulthood was examined in 1,146 African-American adolescent girls and 1,075 Caucasian adolescent girls who completed the Center for Epidemiological Studies of Depression scale. Caucasian girls' scores decreased over time, whereas scores for African-American girls were fairly consistent. Future studies are needed to examine age-specific risk factors in adolescent girls. PMID- 16310135 TI - Understanding and preventing human papillomavirus infection during adolescence and young adulthood. PMID- 16310136 TI - Vaccination as a prevention strategy for human papillomavirus-related diseases. AB - Childhood vaccines have had an enormously beneficial impact on human health. Large-scale vaccination programs have controlled diseases associated with high morbidity and mortality such as poliomyelitis, smallpox, diphtheria, tetanus, yellow fever, pertussis, Haemophilus influenzae, Streptococcus pneumoniae, measles, mumps, and rubella. Human papillomavirus (HPV) is a significant source of morbidity and mortality throughout the world. In the United States, HPV is the most common sexually transmitted infection (STI), and sexually active adolescents are at particularly high risk for HPV acquisition. Genetic and epidemiologic studies have clearly demonstrated that HPV infection is a necessary cause of both cervical cancer and genital warts. More than 99% of cervical cancers contain at least one high-risk HPV type, and approximately 70% contain HPV types 16 or 18. Moreover, low-risk HPV types 6 or 11 are responsible for approximately 90% of genital warts and almost all cases of recurrent respiratory papillomatosis. Thus, a vaccine that could prevent HPV acquisition would have the potential to significantly reduce the incidence of both childhood and adult diseases. PMID- 16310137 TI - Improving adolescent health: focus on HPV vaccine acceptance. AB - The success of future human papillomavirus (HPV) vaccination programs will depend on individuals' willingness to accept vaccination, parents' willingness to have their preadolescent and early adolescent children vaccinated, and health care providers' willingness to recommend HPV vaccination. The purpose of this article is to provide a qualitative review of the relevant literature, including research on knowledge and attitudes about HPV infection and its clinical sequelae, the acceptability of HPV vaccination to individuals and parents, and health care providers' attitudes about recommending HPV vaccination. Additionally, strategies are suggested by which providers of adolescent health care can discuss and recommend HPV vaccines with parents and their children. The research published to date suggests that there is a good deal of misunderstanding about HPV infection, cervical cancer screening, and the sequelae of HPV infection. However, the majority of research studies to date indicate that young women, parents, and health care providers are interested in vaccines that prevent HPV and other sexually transmitted infections (STIs). Of particular note are the consistent findings that providers are less comfortable vaccinating younger versus older adolescents and that endorsement of vaccination by a professional organization is of great importance. Furthermore, research suggests that most parents are interested in having their preadolescent and adolescent children vaccinated against HPV. Parents value the information and recommendations provided by their children's health care providers. To the extent that providers are concerned about potential negative reactions of parents to a recommendation of HPV vaccination, these findings should provide reassurance. At the same time, health care providers will need to be prepared to provide patients and parents with information about HPV and HPV immunization and to respond productively to the rare parent who expresses opposition to HPV vaccine or any other vaccine. PMID- 16310138 TI - Impact of HPV infection in adolescent populations. AB - Human papillomavirus (HPV) is a significant source of morbidity and mortality worldwide. The primary risk factors for acquiring HPV are generally associated with sexual activity. Evidence suggests that condoms provide some protection against infection and disease progression, but any genital contact is sufficient for HPV transmission. HPV is so common and transmissible that having just one sexual partner often results in infection. Indeed, cumulative prevalence rates are as high as 82% among adolescent women in select populations. As such, nearly all sexually active adolescents are at high risk for acquiring HPV. Persistent infection with high-risk HPV types (e.g., HPV 16 or 18) is considered necessary for the development of cervical cancer, whereas infection with low-risk HPV types (e.g., HPV 6 or 11) is associated with the development of genital warts and other low-grade genital abnormalities. Most infections are asymptomatic and are efficiently cleared by the immune system. Similarly, both low- and high-grade lesions caused by HPV can regress in adolescent and young adult women. Treatment guidelines allow for observation of adolescent women who develop low-grade lesions rather than immediate colposcopy. Nonetheless, a small percentage of adolescents will develop precancerous lesions that may progress to invasive cervical cancer. Adolescents should be given appropriate education about HPV and the dangers associated with infection; they should also be encouraged to obtain appropriate gynecological care after initiating sexual activity. This article discusses HPV infection and the causal role that HPV plays in the development of low- and high-grade genital lesions, cervical cancer, and genital warts. PMID- 16310139 TI - A comprehensive picture of palliative care at home from the people involved. AB - The purpose of this study was to identify the comprehensive picture of palliative care in the home, as experienced by the people involved. The study is a secondary analysis of three phenomenological studies including six cancer patients, six next of kin and six district nurses. Data were collected in qualitative interviews using an interview guide. The interviews were tape-recorded and transcribed verbatim. In this secondary analysis, data were analysed by hermeneutic analysis guided by Gadamer. The guiding questions during the reading were: Is there an advantage receiving palliative care at home? Is there a disadvantage receiving palliative care at home? The findings indicate that the advantages of palliative care at home is; striving for normal life, including the care in the home composed of physical care and emotional/mental care. Striving for normal life also includes emotional feelings, safety and resources and policies which regulates this activity. Disadvantages of palliative care at home are commitment, composed of adaptation and extra work, and demands, composed of frustration and uncertainty. If the people involved are to be able to manage the situation and optimize living while dying, there must be support and resources facilitating the situation. PMID- 16310140 TI - Taking the initiative. PMID- 16310141 TI - Q fever-related cholecystitis: a missed entity? PMID- 16310142 TI - High incidence of trachoma in rural areas of Guangxi, China. PMID- 16310144 TI - Favourite papers in infectious diseases 2005. PMID- 16310145 TI - Leaping to extinction? PMID- 16310147 TI - The role of nasal carriage in Staphylococcus aureus infections. AB - Staphylococcus aureus is a frequent cause of infections in both the community and hospital. Worldwide, the increasing resistance of this pathogen to various antibiotics complicates treatment of S aureus infections. Effective measures to prevent S aureus infections are therefore urgently needed. It has been shown that nasal carriers of S aureus have an increased risk of acquiring an infection with this pathogen. The nose is the main ecological niche where S aureus resides in human beings, but the determinants of the carrier state are incompletely understood. Eradication of S aureus from nasal carriers prevents infection in specific patient categories-eg, haemodialysis and general surgery patients. However, recent randomised clinical trials in orthopaedic and non-surgical patients failed to show the efficacy of eliminating S aureus from the nose to prevent subsequent infection. Thus we must elucidate the mechanisms behind S aureus nasal carriage and infection to be able to develop new preventive strategies. We present an overview of the current knowledge of the determinants (both human and bacterial) and risks of S aureus nasal carriage. Studies on the population dynamics of S aureus are also summarised. PMID- 16310148 TI - Treatment options for visceral leishmaniasis: a systematic review of clinical studies done in India, 1980-2004. AB - The state of Bihar in India carries the largest share of the world's burden of antimony-resistant visceral leishmaniasis. We analysed clinical studies done in Bihar with different treatments between 1980 and 2004. Overall, 53 studies were included (all but one published), of which 15 were comparative (randomised, quasi randomised, or non-randomised), 23 dose-finding, and 15 non-comparative. Data from comparative studies were pooled when appropriate for meta-analysis. Overall, these studies enrolled 7263 patients in 123 treatment arms. Adequacy of methods used to do the studies and report on them varied. Unresponsiveness to antimony has developed steadily in the past to such an extent that antimony must now be replaced, despite attempts to stop its progression by increasing dose and duration of therapy. The classic second-line treatments are unsuited: pentamidine is toxic and its efficacy has also declined, and amphotericin B deoxycholate is effective but requires hospitalisation for long periods and toxicity is common. Liposomal amphotericin B is very effective and safe but currently unaffordable because of its high price. Miltefosine-the first oral drug for visceral leishmaniasis-is now registered and marketed in India and is effective, but should be used under supervision to prevent misuse. Paromomycin (or aminosidine) is effective and safe, and although not yet available, a regulatory submission is due soon. To preserve the limited armamentarium of drugs to treat visceral leishmaniasis, drugs should not be deployed unprotected; combinations can make drugs last longer, improve treatment, and reduce costs to households and health systems. India, Bangladesh, and Nepal agreed recently to undertake measures towards the elimination of visceral leishmaniasis. The lessons learnt in Bihar could help inform policy decisions both regionally and elsewhere. PMID- 16310149 TI - Posaconazole: a broad-spectrum triazole antifungal. AB - Posaconazale is a new triazole drug being investigated in phase III clinical trials for the treatment and prevention of invasive fungal infections. In-vitro and in-vivo studies showed that posaconazole has broad-spectrum activity against most Candida species, Cryptococcus neoformans, Aspergillus species, Fusarium species, zygomycetes, and endemic fungi. Posaconazole is given orally two to four times daily. This triazole is widely distributed in the body, metabolised mainly by the liver, and is well tolerated, even in long-term courses. Adverse events are generally mild and include headache and gastrointestinal complaints. Posaconazole has shown promising clinical efficacy against life-threatening fungal infections that are often refractory to the currently available antifungal therapies-eg, invasive aspergillosis, fusariosis, and the emerging zygomycosis. PMID- 16310150 TI - Intravaginal practices, bacterial vaginosis, and women's susceptibility to HIV infection: epidemiological evidence and biological mechanisms. AB - Intravaginal practices such as "dry sex" and douching have been suggested as a risk factor that may increase women's susceptibility to HIV infection. These behaviours appear common in different populations across sub-Saharan Africa, where practices include the use of antiseptic preparations, traditional medicines, or the insertion of fingers or cloths into the vagina. We systematically review the evidence for the association between women's intravaginal practices and HIV infection. Although a number of cross-sectional studies have shown that prevalent HIV infection is more common among women reporting intravaginal practices, the temporal nature of this association is unclear. Current evidence suggests that bacterial vaginosis, which is a likely risk factor for HIV infection, may be a mediator of the association between intravaginal practices and HIV. Although biologically plausible mechanisms exist, there is currently little epidemiological evidence suggesting that intravaginal practices increase women's susceptibility to HIV infection. Further research into factors that increase women's susceptibility to HIV will help to inform the design of vaginal microbicides and other HIV prevention interventions. PMID- 16310151 TI - Intracranial tuberculomas developing while on therapy for pulmonary tuberculosis. AB - We describe the case of a south Asian man who developed symptomatic intracranial tuberculomas while on therapy for pulmonary tuberculosis. The development or progression of intracranial tuberculomas during the course of appropriate antituberculous therapy has been recognised previously. We review the epidemiology, pathophysiology, diagnosis, and treatment of this paradoxical phenomenon. Although the aetiology of this reaction is unknown, it may be related to an enhanced immune response during the course of therapy. Routine brain imaging for all patients diagnosed with tuberculosis is not recommended; however, in patients presenting with new neurological findings, neuroimaging is clearly warranted. Stereotactic brain biopsy should be done whenever the diagnosis of an intracranial tuberculoma is in doubt. When intracranial tuberculomas become symptomatic, antituberculous therapy generally does not need to be changed. Corticosteroids are indicated in all symptomatic cases. PMID- 16310152 TI - A husband and wife with left hand rashes. PMID- 16310153 TI - Application of a human error framework to conduct train accident/incident investigations. AB - Accident/incident investigations are an important qualitative approach to understanding and managing transportation safety. To better understand potential safety implications of recently introduced remote control locomotive (RCL) operations in railroad yard switching, researchers investigated six railroad accidents/incidents. To conduct the investigations, researchers first modified the human factors analysis and classification system (HFACS) to optimize its applicability to the railroad industry (HFACS-RR) and then developed accident/incident data collection and analysis tools based on HFACS-RR. A total of 36 probable contributing factors were identified among the six accidents/incidents investigated. Each accident/incident was associated with multiple contributing factors, and, for each accident/incident, active failures and latent conditions were identified. The application of HFACS-RR and a theoretically driven approach to investigating accidents/incidents involving human error ensured that all levels of the system were considered during data collection and analysis phases of the investigation and that investigations were systematic and thorough. Future work is underway to develop a handheld software tool that incorporates these data collection and analysis tools. PMID- 16310154 TI - Evaluation of variable speed limits for real-time freeway safety improvement. AB - Use of various variable speed limit (VSL) strategies as a tool for safety improvement on freeways was evaluated using simulation of a section of Interstate 4 in Orlando, FL. Real-time crash likelihood was calculated based on models developed in previous research by the first author [Abdel-Aty, M., Uddin, N., Pande, A., January 2005. Split models for predicting multi-vehicle crashes during high-speed and low-speed operating conditions on freeways. In: Presented at the 84th Annual Meeting of the Transportation Research Board, Washington, DC]. VSL implementation produced safety improvement by simultaneously implementing lower speed limits upstream and higher speed limits downstream of the location where crash likelihood is observed in real-time. This improvement was realized in the case of medium-to-high-speed regimes on the freeway, but no benefit was achieved in low-speed situations (no substantial safety benefit from implementing VSL in congested situation's simulation). The final recommendations for implementing VSL are: gradually introducing speed limit changes over time (5 mph every 10 min); abruptly changing speed limit in space ( no gap distance); reducing speed limits upstream and increasing speed limits downstream of location of interest; the speed limit changes up- and downstream should be large in magnitude (15 mph) and implemented within short distances (2 miles) of the location of interest. In addition to the safety benefit, this final strategy also produced travel time savings. PMID- 16310155 TI - The nature of residual cost in regular switch response factors. AB - Two experiments are presented that compare the residual cost found when switching from one task to another under predictable conditions. The aim of the study was to explore the roles played by the stimulus, the response, or both in the process of the mental set reconfiguration necessary to switch between two tasks. The experiments tested [Rogers, R. D., & Monsell, S. (1995). Cost of a predictable switch between simple cognitive tasks. Journal of Experimental Psychology: General, 124, 207-231] stimulus-cued-completion hypothesis and [Schuch, S., & Koch, I. (2003). The role of response selection for inhibition of task sets in task shifting. Journal of Experimental Psychology: Human Perception and Performance, 29, 92-105] hypothesis of response selection as the key factor in the nature of switch cost. In the first experiment, two conditions were created that varied in terms of a Go/No-Go signal: The Go trials were a replication of [Tornay, F. J., & Milan, E. G. (2001). A more complete task-set reconfiguration in random than in predictable task switch. The Quarterly Journal of Experimental Psychology, 54A, 785-803 Experiment 3]; The No-Go trials were identical to the first condition, except that participants did not execute a response in the trial n-1 (Schuch & Koch, 2003). In addition, the percentage of Go and No-Go trials was manipulated. The results showed that the cost was significant only in the high Go signal-frequency case (Experiment 2), with an abrupt offset in Go trials and a gradual offset in No-Go trials. Based on the results of these experiments, it was concluded that the crucial factor to complete a mental set reconfiguration is response-related and not stimulus-related. PMID- 16310156 TI - Plasmodium vivax dihydrofolate reductase point mutations from the Indian subcontinent. AB - Mutations in Dihydrofolate Reductase (dhfr) gene of Plasmodium vivax are known to be associated with resistance to antifolate drugs. To analyze the extent of these mutations in P. vivax population in India, dhfr gene was isolated and sequenced for 121 P. vivax isolates originating from different geographical regions of Indian subcontinent. These sequences were compared with the gene sequence that represent wild type sequence (accession no. X98123). P. vivax dhfr (Pvdhfr) sequences showed limited polymorphism and about 70% isolates showed wild type dhfr sequence. A total of 36 mutations were found at 11 positions in 121 isolates. A majority of mutant isolates showed double mutations at residues 58 (S ->R) and 117 (S-->N), known to be associated with pyrimethamine resistance, but only 19% showed double mutations at residues 57 (F-->L) and 58 (S-->R). Pvdhfr alleles showing quadruple mutation (F57L, S58R, T61M and S117T) were found in two isolates. Three other mutations reported earlier at residue 13, 33 and 173 were not found in any of the Isolates. Six novel mutations at residues 38 (R-->G), 93 (S-->C), 109 (S-->H), 131 (R-->G), 159 (V-->A) and 188 (I-->V) were observed in seven isolates. Whether these novel mutations are linked to pyrimethamine resistance remains to be established. PMID- 16310157 TI - Binding mitochondria to cryogel monoliths allows detection of proteins specifically released following permeability transition. AB - Following proapoptotic signals such as calcium-induced mitochondrial permeability transition or translocation of proapoptotic proteins, mitochondria induce cell death through release of apoptogenic proteins. The mechanism of release and the identity of the released proteins are currently debated. Earlier attempts at identification of the apoptogenic proteins have been hampered by a high nonspecific background. Our aim was to develop a novel method where background release was eliminated, allowing proteins specifically released from mitochondria following proapoptotic stimulation to be identified. Liver mitochondria were immobilized and washed on cryogel monoliths prior to induction of protein release (calcium or Bid/Bax). Immobilized mitochondria exhibited normal morphology and swelling response and retained respiratory activity. The released proteins were collected, concentrated, separated on polyacrylamide gels which were cut into pieces, trypsin-digested, and analyzed using liquid chromatography-tandem mass spectrometry. Control samples contained no protein, and stimulation with calcium and Bid/Bax resulted in identification of 68 and 82 proteins, respectively. We conclude that, in combination with the robust proteomic approach, immobilization on cryogel monoliths is a fruitful approach for studying specific protein release from isolated mitochondria. We propose that this method is a powerful tool to further characterize the role of mitochondria in cell death induction. PMID- 16310158 TI - [Aberrant pancreas and duodenal diaphragm: an exceptional association]. AB - Aberrant pancreas of the duodenal wall (APD) and duodenal diaphragm (DD) are two rare entities, which developed during duodenal embryogenesis. Occurrence, diagnosis and therapeutic approach of these lesions are different. Herein, we report the first case of this exceptional association in a man who had no symptoms. A surgical resection of both lesions was performed and the outcome was uneventful. Embryogenesis, morphological characteristics and treatment are discussed. PMID- 16310159 TI - [Improvement of medical results and of cost/efficiency needs periodic assessment of practice]. PMID- 16310160 TI - The sterol-binding antibiotic nystatin inhibits entry of non-opsonized Leishmania donovani into macrophages. AB - Leishmania donovani is an obligate intracellular parasite that infects macrophages of the vertebrate host resulting in visceral leishmaniasis in humans, a major public health problem worldwide. The molecular mechanisms involved in internalization of Leishmania are still poorly characterized. We report here that cholesterol sequestration by the sterol-binding antifungal polyene antibiotic nystatin markedly inhibits binding and entry of non-opsonized L. donovani promastigotes into macrophages. Interestingly, these effects are not observed when serum-opsonized L. donovani are used for infectivity studies thus pointing the essential role of cholesterol in mediating entry of the parasite via the non opsonic pathway. Based on our earlier results where leishmanial infectivity was shown to be sensitive to physical depletion of cholesterol from macrophages, these results indicate that the mere sequestration of cholesterol in the host plasma membrane is sufficient to inhibit the binding and entry of non-opsonized L. donovani. These results represent the first report on the effect of a cholesterol-sequestering agent on the entry of Leishmania parasites to host macrophages. More importantly, these findings offer the possibility of reevaluating the mechanism behind the effectiveness of current therapeutic strategies to treat leishmaniasis. PMID- 16310161 TI - Altered expression and function of sodium channels in large DRG neurons and myelinated A-fibers in early diabetic neuropathy in the rat. AB - Differential alterations of sodium channels in small nociceptive C-fiber DRG neurons have been implicated in diabetic neuropathy. In this study, we investigated sodium currents and the expression of sodium channels in large A fiber DRG neurons in diabetic rats. Compared with controls, large neurons from diabetic rats showed significant increases in both total and TTX-S sodium currents and approximately -15mV shifts in their voltage-dependent activation kinetics. TTX-R Na(v)1.9 sodium current was also significantly increased, whereas no alteration of TTX-R Na(v)1.8 current was observed in neurons from diabetic rats. Sodium current induced by fast- or slow-voltage ramps increased markedly in the diabetic neurons as well. Immunofluorescence studies showed significant increases in the levels and number of large DRG neurons from diabetic rats expressing Na(v)1.2, Na(v)1.3, Na(v)1.7, and Na(v)1.9 whereas Na(v)1.8 decreased. We also observed a decrease in the number of nodes of Ranvier expressing Na(v)1.8 and in staining intensity of Na(v)1.6 and Na(v)1.8 at nodes. Our results suggest that alterations of sodium channels occur in large DRG neurons and A-fibers, and may play an important role in diabetic sensory neuropathy. PMID- 16310162 TI - Symmetric array of the urothelium surface controlled by the lipid lattice composition. AB - Proteins on the luminal surface of the mammalian membrane urothelium form an hexagonal two-dimensional (2D) lattice of approximately 12nm particles embedded in a lipid matrix. The establishment and/or maintenance of this two-dimensional lattice have not yet been elucidated. To understand the lipid involvement in these effects, which may have a potential role on the permeability barrier, the lattice structural changes induced by different dietary fatty acids were analyzed. Image analysis of the uranyl formate stained urothelial membrane from mice fed with a commercial diet (control) or a formula containing 5% (w/w) corn oil (CO) or olein (O) and our previous data of fluorescence anisotropy indicated that the olein diet profoundly alters the two-dimensional particle network. Besides the increase from 15nm (control) to 17nm center-to-center particle, a statistically significant increase of the particle size was induced by the olein diet. The same parameters on urothelial membranes from animals fed with a corn diet differed on the long-range super-array from the control values. A specific structural change of the urothelial plaque particle organization as a response to changes of lipid composition is described for the first time. These results are a starting point for further research of the lipid implications on the permeability barrier. PMID- 16310163 TI - Thrombospondin 1 mediates angiotensin II induction of TGF-beta activation by cardiac and renal cells under both high and low glucose conditions. AB - Renal and cardiac fibrosis leading to organ failure are complications of both diabetes and hypertension. These disease processes, when combined, exacerbate development of fibrotic complications. Control of latent transforming growth factor (TGF)-beta activation is a potential determinant of fibrotic progression. Both glucose and angiotensin II (Ang II) upregulate thrombospondin-1 (TSP1), a major activator of latent TGF-beta, and stimulate increased TGF-beta activity. We previously showed that high glucose stimulated TSP1-dependent TGF-beta activation in rat mesangial cells (RMCs). In this paper, we examined whether Ang II similarly upregulates TSP1 production and TSP1-dependent TGF-beta activation alone or in combination with high glucose concentrations. Ang II and high glucose stimulated increases in TSP1 protein levels in the conditioned media of both rat cardiac fibroblasts (RCFs) and rat mesangial cells (RMCs). Meanwhile, Ang II stimulated increases in both TGF-beta activity and protein by RMCs, whereas, RCFs responded to both Ang II and high glucose with increased TGF-beta activity in the absence of altered TGF-beta protein levels. A combination of Ang II and high glucose induced synergistic TGF-beta activation by RCFs. Moreover, Ang II induction of TSP1 and increased TGF-beta activity were blocked by losartan, an antagonist of the Ang II type 1 (AT1) receptor. The increase in TSP1 expression leads to increased TGF-beta activity upon Ang II and/or glucose treatment, since peptide antagonists of TSP1-mediated TGF-beta activation blocked Ang II and glucose-induced TGF-beta activation. Our data support a role for TSP1 in the development and progression of renal and cardiac fibrosis in hypertension and diabetes. PMID- 16310164 TI - Oxidative stress provokes atherogenic changes in adipokine gene expression in 3T3 L1 adipocytes. AB - Increased oxidative stress has been associated with obesity-related disorders. In this study, we investigated how oxidative stress, in different ways of exposure, regulates gene expression of various adipokines in 3T3-L1 adipocytes. Exposure to 100-500microM H(2)O(2) for 10min, as well as exposure to 5-25mU/ml glucose oxidase for 18h, similarly decreased adiponectin, leptin, and resistin mRNAs, and increased plasminogen activator inhibitor-1 mRNA. Secretion levels of adipokines were also changed by oxidative stress in parallel with mRNA expression levels. Although a peak increase in plasminogen activator inhibitor-1 mRNA was achieved between 4 and 8h after exposure to H(2)O(2) for 10min, significant decreases in adiponectin and resistin mRNA were observed after 16h, while leptin mRNA was decreased earlier. Our results suggest that oxidative stress, even of short duration, has a significant impact on the regulation of various adipokine gene expressions favoring atherosclerosis. PMID- 16310165 TI - Cellular entry of Hantaan virus A9 strain: specific interactions with beta3 integrins and a novel 70kDa protein. AB - Cellular entry of pathogenic hantaviruses had been shown to be mediated by beta3 integrins. However, no direct evidence exists that hantavirus binds to beta3 integrins, and integrin beta3 subunit is not expressed on some cells permissive to hantavirus infection. In this report, utilizing beta3-integrin-transfected CHO cells, we demonstrated that integrin beta3 subunit renders CHO cells susceptible to Chinese Hantaan virus (HTN) strain A9 (isolated in China), and the viral infection was correspondingly inhibited by antibodies to alphavbeta3, alphaIIbbeta3, beta3, and alphav integrins. Furthermore, virus overlay protein binding assay and 'quarternary Western' analysis indicate that HTN A9 directly interacts with beta3 integrins and an unidentified 70kDa protein. These findings indicate that beta3 integrins play a crucial role in cellular entry of HTN A9 via specific interactions with the virus. In addition, a novel 70kDa protein may serves as a candidate receptor or alternative cellular component for interaction with HTN. PMID- 16310166 TI - Correlation between mRNA and protein abundance in Desulfovibrio vulgaris: a multiple regression to identify sources of variations. AB - Parallel profiling of mRNA and protein on a global scale and integrative analysis of these two data types could provide additional insights into the metabolic mechanisms underlying complex biological systems. However, because mRNA and protein abundance are affected by many cellular and physical processes, there have been conflicting results on their correlation. Using whole-genome microarray and LC-MS/MS proteomic data collected from Desulfovibrio vulgaris grown under three different conditions, we systematically investigate the relationship between mRNA and protein abundance by a multiple regression approach, in which some of the key covariates that may affect mRNA-protein relationship were included. The results showed that mRNA abundance alone can explain only 20-28% of the total variation of protein abundance, suggesting mRNA-protein correlation can not be determined by mRNA abundance alone. Among various covariates, analytic variation of protein abundance is the major source for the variation of mRNA protein correlation, which contributes to 34-44% of the total variation of mRNA protein correlation. The cellular functional category of genes/proteins contributes 10-15% of the total variation of mRNA-protein correlation, with a more pronounced correlation of the two properties was observed for "central intermediary metabolism" and "energy metabolism" categories. In addition, protein stability also contributes 5% of the total variation of mRNA-protein correlation. The study presents the first quantitative analysis of the contributions of various biochemical and physical sources to the correlation of mRNA and protein abundance in D. vulgaris. PMID- 16310167 TI - Enzymatic synthesis of heparin related polysaccharides on sensor chips: rapid screening of heparin-protein interactions. AB - The biological roles of heparin (HP) and heparan sulfate (HS) are mediated mainly through their interaction with proteins. In the present work, we provide a rapid method for screening HP/HS-protein interactions providing structural data on the key sulfo groups that participate in the binding. A library of polysaccharides structurally related to HP was prepared by immobilizing the biotinylated N sulfated K5 polysaccharide (N-sulfoheparosan) on sensor chips followed by selective modification of this polysaccharide with enzymes that participate in HP/HS biosynthesis. The polysaccharides synthesized on the surface of the sensor chips differ in the number and position of sulfo groups present both on uronic acid and glucosamine residues. Surface plasmon resonance was used to measure the interaction of each member of this polysaccharide library with antithrombin III (ATIII), to afford structural information on sulfo groups required for this HP/HS protein interaction. This method is viewed as widely applicable for the study of the structure-activity relationship (SAR) of HP/HS-protein interactions. PMID- 16310168 TI - Rapid assessment of two major repair activities against DNA double-strand breaks in vertebrate cells. AB - A linearized plasmid DNA, in which tandem repeats of 400bp flank the breakpoints, was transfected into vertebrate cells, and breakpoint junctions of plasmid DNA circularized in the cells were analyzed to assess the repair activities against DNA double-strand break (DSB) by non-homologous end joining and homology-directed repair (i.e., homologous recombinational repair and single-strand annealing). The circularization by non-homologous end joining repair of the breakpoints depended on the expression of DNA-PKcs, while that by homology-directed repair through the repeats depended on the length of the repeats, indicating that these two DSB repair activities can be rapidly assessed by this assay. Predominance in circularization by either non-homologous end joining or homology-directed repair differed among cells examined, and circularization was exclusively undertaken by homology-directed repair in DT40 cells known to show a high homologous recombination rate against gene-targeting vectors. Thus, this assay will be helpful in studies on mechanisms and inter-cellular variations of DSB repair. PMID- 16310169 TI - Influence of heparin and dendrimers on the aggregation of two amyloid peptides related to Alzheimer's and prion diseases. AB - Amyloid plaques composed of proteinaceous aggregates are commonly found in brains affected by Alzheimer's disease and spongiform encephalopaties. A structural homology has been recently described for the Alzheimer's peptide Abeta1-28 and the segment of the prion protein Prp185-208. In the present paper, further elements in common are reported: the aggregation processes are in both cases enhanced by the model glucosaminoglycan heparin and dendrimers can modulate the aggregation process by affecting the nucleation rate at low concentrations and the elongation rate at high concentrations. Nucleation and elongation rate constants are derived from fittings to a nucleation dependent polymerization model. PMID- 16310170 TI - Interleukin-1beta-induced apoptosis through adenylyl cyclase and ERK1/2 inhibition in primary cultured thyroid cells. AB - The programmed cell death plays a crucial role in the regulation of numerous physiological and pathological phenomena. In this study, we show that interleukin 1 beta (IL-1beta) induces an early production of endogenous ceramides via N sphingomyelinase (N-Smase) as well as an inhibition of adenylyl cyclase activity in pig thyroid cells. This effect is followed by a down-regulation of the extracellular signal-regulated protein kinase (ERK1/2) phosphorylation, an activation of caspase-3, and ends by setting up the programmed cell death. The permeable exogenous C(2)-ceramide reproduces IL-1beta effects on: (i) inhibition of adenylyl cyclase activity, (ii) down-regulation of ERK1/2 phosphorylation, (iii) activation of caspase-3, and (iv) apoptosis in pig thyroid cells. Cell treatment with a PKA inhibitor down-regulates ERK1/2 phosphorylation. Furthermore, inhibition of ERK1/2 signaling pathway by U-0126 enhances caspase-3 activity and sets up programmed cell death. Both IL-1beta and exogenous C(2) ceramide effects are reproduced by U-0126 so illustrating the implication of ERK1/2 down-regulation in both caspase-3 activation and apoptosis induction. Our study shows for the first time that endogenous ceramides are important second messengers in IL-1beta-induced apoptosis in pig thyroid cells through inhibition of adenylyl cyclase and ERK1/2 activities. PMID- 16310171 TI - Interactions of low-molecular-weight semi-synthetic sulfated heparins with human leukocyte elastase and human Cathepsin G. AB - Semi-synthetic low-molecular-weight heparin samples (LMWHs), having homogeneous degree of polymerization and saccharide backbone, but differing in the number and location of sulfate groups, were investigated in their ability to interfere with the pharmacologically relevant targets human leukocyte elastase (EL) and human Cathepsin G (CatG). Spectroscopic studies were performed for a quantitative evaluation of the enzyme-inhibitor dissociation constant, K(i), and of the IC(50) values for the inhibition of cleavage of target peptide sequences. Both proteases are inhibited by the tested polysaccharides through a mixed hyperbolic binding process. A non-linear relationship was found between degree of sulfation and binding affinity or enzyme inhibition properties, showing a composite correlation between heparin charge density and interference with EL/CatG activity. PMID- 16310172 TI - 6-Benzylthioinosine analogues: promising anti-toxoplasmic agents as inhibitors of the mammalian nucleoside transporter ENT1 (es). AB - Certain 6-benzylthioinosine analogues have been identified as potential chemotherapeutic agents against Toxoplasma gondii in cell culture and animal models. These compounds are selectively transported and metabolized by toxoplasma infected, but not uninfected, cells. In sharp contrast to mammalian nucleoside transporters, the toxoplasma adenosine/purine transporter (TgAT) allows the transport of these 6-benzylthioinosine analogues into infected cells. After entering the infected cell, these compounds act as subversive substrates for toxoplasma, but not the host, adenosine kinase (EC.2.7.1.20). Hence, 6 benzylthioinosine analogues become toxic to toxoplasma infected cells, but not uninfected host cells or animals. The basis for the lack of uptake of the anti toxoplasmic 6-benzylthioinosines by uninfected host cells is currently unknown. These anti-toxoplasmic 6-benzylthioinosines may not be substrates for the mammalian nucleoside transporters or they may act as inhibitors of these transporters. Previous studies have shown that some 6-benzylthioinosines are inhibitors of the mammalian nucleoside transporter ENT1 (es). Therefore, we examined the efficacy of promising anti-toxoplasmic 6-benzylthioinosines as inhibitors of ENT1 (es) in an effort to elucidate the basis for the lack of uptake of anti-toxoplasmic 6-benzylthioinosines by uninfected host cells. The results showed that these compounds are inhibitors of ENT1 (es). In general, electron-withdrawing substituents at the ortho, meta or para positions of the benzyl ring improved binding. The most potent inhibitors identified were m- and p nitro-6-benzylthioinosine, which had K(i) values in the subnanomolar range. Therefore, anti-toxoplasmic 6-benzylthioinosines are not only selectively toxic to parasites and parasite infected cells, they are also inhibitors of nucleoside transport in host cells. This inhibition of the host nucleoside transport is an added advantage for these 6-benzylthioinosine analogues as anti-toxoplasmic agents. Inhibitors of nucleoside transport in mammalian cells can selectively protect the host from the toxicity of toxic purine nucleosides that may be used in future combination therapy against toxoplasmosis or from metabolites of the 6 benzylthioinosine analogues that may be released by the destruction of infected cells. These findings further advance the rationale for developing 6 benzylthioinosine analogues as selective therapeutic agents for the treatment of toxoplasmosis. PMID- 16310173 TI - Mobilization of iron from cells by hydroxyquinoline-based chelators. AB - With the aim of identifying an iron (Fe) chelator which is effective at mobilizing intracellular Fe, two novel ligands were synthesized and tested. Hydroxyquinoline is known to possess a high affinity for Fe and was thus chosen as the Fe binding motif for the hexadentate chelators, C1 (2,2'-[ethane-1,2 diylbis(iminomethylene)]diquinolin-8-ol) and C2 (2,2'-[cyclohexane-1,2 diylbis(iminomethylene)]diquinolin-8-ol). Both chelators are lipophilic, with Fe3+ complexes slightly more hydrophilic than the free ligands. C1 and C2 were equally toxic to K562 cells, and partial protection was afforded by supplementing the culture medium with human holotransferrin, suggesting that some of the toxicity of the ligands is due to cellular Fe depletion. Micromolar concentrations of both ligands effectively mobilized 59Fe from reticulocytes and K562 cells. In reticulocytes, 50 microM C1 caused the release of 60% of the cells' initial 59Fe uptake after a 4h incubation. Under the same conditions, C2 revealed a release of 50% of the 59Fe. Overall, both ligands merit in vivo study for oral activity. Their effectiveness at low concentrations makes them candidates for therapeutic use. PMID- 16310174 TI - Parametrical modelling of a premature ventricular contraction ECG beat: comparison with the normal case. AB - The aim of this paper is to analyse a parametrical Gaussian kernel based model. The proposed model is tested on two types of electrocardiogram (ECG) beats, the normal case beat and the premature ventricular contraction (PVC) one. Basically, the model is constituted of N Gaussians where their corresponding parameters are estimated by optimising a specific criterion. The modelling technique has been validated using MIT/BIH databases. As a result of this study, we show that a normal beat can be modelled using 18 parameters and only 15 parameters are needed to reconstruct the PVC one. PMID- 16310175 TI - Non-equivalence of embryonic and somatic cell nuclei affecting spindle composition in clones. AB - Cloning by nuclear transfer remains inefficient but is more efficient when nuclei from embryonic cells or embryonic stem cells (ECNT) are employed as compared with somatic cells (SCNT). The factors determining efficiency have not been elucidated. We find that somatic and embryonic nuclei differ in their ability to organize meiotic and mitotic spindles of normal molecular composition. Calmodulin, a component of meiotic and mitotic spindle chromosome complexes (SCCs), displays sharply reduced association with the SCC forming after SCNT but not ECNT. This defect persists in mitotic spindles at least through the second mitosis, despite abundant calmodulin expression in the cell, and correlates with slow chromosome congression. We propose that somatic cell nuclei lack factors needed to direct normal SCC formation in oocytes and early embryos. These results reveal a striking control of SCC formation by the transplanted nucleus and provide the first identified molecular correlate of donor stage-dependent restriction in nuclear potency. PMID- 16310176 TI - A rat 8 kb dentin sialoprotein-phosphophoryn (DSP-PP) promoter directs spatial and temporal LacZ activity in mouse tissues. AB - Dentin sialoprotein (DSP) and phosphophoryn (PP) are two major dentin noncollagenous proteins that are encoded on a single DSP-PP transcript whose expression is tightly regulated during tooth dentinogenesis. The recent identification of this gene transcript in other tissues, including inner ear and jaw tissue, suggests that DSP and PP may have pleiotropic effects on other organs besides teeth. To identify candidate regulatory elements that control DSP-PP temporal and spatial expression, we constructed a -5 kb upstream region rat DSP PP promoter into the beta-galactosidase expression vector pnLacF plasmid and used this construct to prepare DSP-PP-LacZ transgenic mice. Multiple mouse tissues including teeth, bone, and kidney obtained from the six resulting transgenic mouse lines displayed strong LacZ activity. This spatial distribution was confirmed in several of these tissues by in situ hybridization studies. LacZ activity was transiently expressed in preameloblasts and continuously expressed in odontoblasts demonstrating that this -5 kb rat promoter-dependent LacZ expression mimics reported DSP-PP mRNA expression patterns. Interestingly, this 5 kb rat promoter construct drives LacZ expression according to the rat developmental clock. Based on identified transcription factors present in this -5 kb promoter region, we have identified several probable cis-regulatory modules whose interaction with one another could account for the spatial and temporal distribution of DSP-PP transcripts in developing tissues. PMID- 16310177 TI - Fishing for prostanoids: deciphering the developmental functions of cyclooxygenase-derived prostaglandins. AB - Prostaglandin G/H synthases (PGHS), commonly referred to as cyclooxygenases (COX 1 and COX-2), catalyze a key step in the synthesis of biologically active prostaglandins (PGs), the conversion of arachidonic acid (AA) into prostaglandin H(2) (PGH(2)). PGs have important functions in a variety of physiologic and pathologic settings, including inflammation, cardiovascular homeostasis, reproduction, and carcinogenesis. However, an evaluation of prostaglandin function in early development has been difficult due to the maternal contribution of prostaglandins from the uterus. The emergence of zebrafish as a model system has begun to provide some insights into the roles of this signaling cascade during vertebrate development. In zebrafish, COX-1 derived prostaglandins are required for two distinct stages of development, namely during gastrulation and segmentation. During gastrulation, PGE(2) signaling promotes cell motility, without altering the cell shape or directional migration of gastrulating cells. During segmentation, COX-1 signaling is also required for posterior mesoderm development, including the formation of vascular tube structures, angiogenesis of intersomitic vessels, and pronephros morphogenesis. We propose that deciphering the role for prostaglandin signaling in zebrafish development could yield insight and ultimately address the mechanistic details underlying various disease processes that result from perturbation of this pathway. PMID- 16310178 TI - Stem cells and their derivatives can bypass the requirement of myocardin for smooth muscle gene expression. AB - The Serum Response Factor (SRF) coactivator myocardin stimulates the transcription of multiple muscle genes during cardiac and smooth muscle development. Mouse embryos lacking myocardin die during the earliest stages of smooth muscle development and fail to express multiple smooth muscle marker genes in the embryonic dorsal aorta and other vascular structures. In this study, we used mutant embryonic stem cell lines to further define the role of myocardin in smooth muscle differentiation and vascular development. Misexpression of myocardin in undifferentiated muscle stem cells resulted in efficient activation of smooth muscle genes, and weaker activation of genes involved in cardiac and skeletal muscle differentiation. Remarkably, myocardin(-/-) embryonic stem cell lines differentiated into smooth muscle cells in vitro, although these cells expressed significantly decreased levels of smooth muscle contractile genes. Moreover, genetically labeled myocardin(-/-) ES cells were able to contribute to smooth muscle lineages in vivo. These results indicate that while myocardin function is sufficient for activation of SRF-dependent muscle gene expression in multiple cell types, myocardin-independent mechanism(s) can suffice for expression in some smooth muscle lineages. PMID- 16310179 TI - Dazl deficiency leads to embryonic arrest of germ cell development in XY C57BL/6 mice. AB - Genes of the DAZ family play critical roles in germ cell development in mammals and other animals. In mice, Dazl mRNA is first observed at embryonic day 11.5 (E11.5), but previous studies using Dazl-deficient mice of mixed genetic background have largely emphasized postnatal spermatogenic defects. Using an inbred C57BL/6 background, we show that Dazl is required for embryonic development and survival of XY germ cells. By E14.5, expression of germ cell markers (Mvh, Oct4, Dppa3/Stella, GCNA and MVH protein) was reduced in XY Dazl-/- gonads. By E15.5, most remaining germ cells in XY Dazl-/- embryos exhibited apoptotic morphology, and XY Dazl-/- gonads contained increased numbers of TUNEL positive cells. The rare XY Dazl-/- germ cells that persisted until birth maintained a nuclear morphology that resembled that of wildtype germ cells at E12.5-E13.5, a critical developmental period when XY germ cells lose pluripotency and commit to a spermatogonial fate. We propose that Dazl is required as early as E12.5-E13.5, shortly after its expression is first detected, and that inbred Dazl /- mice of C57BL/6 background provide a reproducible standard for exploring Dazl's roles in embryonic germ cell development. PMID- 16310180 TI - Genome-wide expression dynamics during mouse embryonic development reveal similarities to Drosophila development. AB - Gene transcription mediates many vital aspects of mammalian embryonic development. A comprehensive characterization and analysis of the dynamics of gene transcription in the embryo is therefore likely to provide significant insights into the basic mechanisms of this process. We used microarrays to map transcription in the mouse embryo in the important period from embryonic day 8 (e8.0) to postnatal day 1 (p1) during which the bulk of the differentiation and development of organ systems takes place. Analysis of these expression profiles revealed distinct patterns of gene expression which correlate with the differentiation of organs including the nervous system, liver, skin, lungs, and digestive system, among others. Statistical analysis of the data based on Gene Ontology (GO) group annotation showed that specific temporal sequence patterns in gene class utilization across development are very similar to patterns seen during the embryonic development of Drosophila, suggesting conservation of the temporal progression of these processes across 550 million years of evolution. The temporal profiles of gene expression and activation of processes revealed here provide intriguing insights into the mechanisms of mammalian development, embryogenesis, and organogenesis, as well as into the evolution of developmental processes. PMID- 16310181 TI - Inhibition of acute nociceptive responses in rat spinal cord by a bradykinin B1 receptor antagonist. AB - This study used behavioural and in vivo electrophysiological paradigms to examine the effects of systemic and spinal administration of a bradykinin B1 receptor antagonist, compound X, on acute nociceptive responses in the rat. In behavioural experiments, compound X significantly increased the latency to withdraw the hindpaw from a radiant heat source after both intravenous and intrathecal administration, without affecting motor performance on the rotarod. In electrophysiological experiments, both intravenous and direct spinal administration of compound X attenuated the responses of single dorsal horn neurones to noxious thermal stimulation of the hindpaw. These data show that the antinociceptive effects of a bradykinin B1 receptor antagonist are mediated, at least in part, at the level of the spinal cord and suggest a role for spinal bradykinin B1 receptors in acute nociception. PMID- 16310182 TI - The ruthenium-based nitric oxide scavenger, AMD6221, augments cardiovascular responsiveness to noradrenaline in rats with streptozotocin-induced diabetes. AB - Excess production of nitric oxide by inducible nitric oxide synthase (iNOS) has been implicated in cardiovascular dysfunction associated with the acute phase of diabetes mellitus. We examined if the selective nitric oxide scavenger, AMD6221 (ruthenium[hydrogen(diethylenetrinitrilo)pentaacetato] chloride) improved cardiovascular function in rats with streptozotocin (60 mg/kg, i.v.)-induced diabetes. The cardiovascular effects of noradrenaline (16.5 nmol/kg/min, i.v.) were measured in thiobutabarbitone-anaesthetised diabetic and control rats before and after acute administration of AMD6221 (80 mg/kg). Rats in the acute phase of diabetes (3 weeks post injection of streptozotocin) had impaired mean arterial pressure, left ventricular systolic pressure and maximum rate of increase (+dP/dt) and decrease (-dP/dt) of left ventricular pressure responses to noradrenaline compared with control rats. AMD6221 significantly augmented noradrenaline-induced increases in left ventricular systolic pressure and +/ dP/dt in the diabetic but not control rats. The results show that selective scavenging of nitric oxide by AMD6221 improved cardiac response to noradrenaline in rats with streptozotocin-induced diabetes. PMID- 16310183 TI - 5-HT1A and 5-HT1B receptor agonists and aggression: a pharmacological challenge of the serotonin deficiency hypothesis. AB - More than any other brain neurotransmitter system, the indolamine serotonin (5 HT) has been linked to aggression in a wide and diverse range of species, including humans. The nature of this linkage, however, is not simple and it has proven difficult to unravel the precise role of this amine in the predisposition for and execution of aggressive behavior. The dogmatic view that 5-HT inhibits aggression has dominated both pharmacological research strategies to develop specific and effective novel drug treatments that reduce aggressive behavior and the pharmacological mechanistic interpretation of putative serenic drug effects. Our studies on brain serotonin and aggression in feral wild-type rats using the resident-intruder paradigm have challenged this so-called serotonin deficiency hypothesis of aggressive behavior. The well-known fact that certain 5-HT(1A/1B) receptor agonists potently and specifically reduce aggressive behavior without motor slowing and sedative effects is only consistent with this hypothesis under the assumption that the agonist mainly acts on the postsynaptic 5-HT(1A/1B) receptor sites. However, systemic injections of anti-aggressive doses of 5-HT(1A) and (1B) agonists robustly decrease brain 5-HT release due to their inhibitory actions at somatodendritic and terminal autoreceptors, respectively. The availability of the novel benzodioxopiperazine compound S-15535, which acts in vivo as a preferential agonist of the somatodendritic 5-HT(1A) auto-receptor and as an antagonist (weak partial agonist) at postsynaptic 5-HT(1A) receptors, allows for a pharmacological analysis of the exact site of action of this anti aggressive effect. It was found that, similar to other prototypical full and partial 5-HT(1A) and/or 5-HT(1B) receptor agonists like repinotan, 8-OHDPAT, ipsapirone, buspirone, alnespirone, eltoprazine, CGS-12066B and CP-93129, also S 15535 very effectively reduced offensive aggressive behavior. Unlike the other ligands, however, a remarkable degree of behavioral specificity was observed after treatment with S-15535, in that the anti-aggressive effects were not accompanied by inhibiting (like other 5-HT(1A) receptor agonist with moderate to high efficacy at postsynaptic 5-HT(1A) receptors) or enhancing (like agonists with activity at 5-HT(1B) receptors and alnespirone) non-aggressive motor behaviors (e.g., social exploration, ambulation, rearing, and grooming) beyond the range of undrugged animals with corresponding levels of aggression. The involvement of 5-HT(1A) and/or 5-HT(1B) receptors in the anti-aggressive actions of these drugs was convincingly confirmed by showing that the selective 5-HT(1A) receptor antagonist WAY-100635 and/or the 5-HT(1B) receptor antagonist GR-127935, while inactive when given alone, effectively attenuated/prevented these actions. Furthermore, combined administration of S-15535 with either alnespirone or CGS 42066B elicited a clear additive effect, indicated by a left-ward shift in their dose-effect curves, providing further support for presynaptic sites of action (i.e., inhibitory somatodendritic 5-HT(1A) and terminal 5-HT(1B) autoreceptors). These findings strongly suggest that the specific anti-aggressive effects of 5 HT(1A) and 5-HT(1B) receptor agonists are predominantly based on reduction rather than enhancement of 5-HT neurotransmission during the combative social interaction. Apparently, normal display of offensive aggressive behavior is positively related to brief spikes in serotonergic activity, whereas an inverse relationship probably exists between tonic 5-HT activity and abnormal forms of aggression only. PMID- 16310184 TI - Inhibition of 1-methyl-4-phenylpyridinium-induced mitochondrial dysfunction and cell death in PC12 cells by sulfonylurea glibenclamide. AB - The present study investigates the effect of sulfonylurea glibenclamide on the cytotoxicity of 1-methyl-4-phenylpyridinium (MPP+) in differentiated PC12 cells in relation to changes in the mitochondrial membrane permeability. Glibenclamide and tolbutamide reduced the MPP+-induced cell death and GSH depletion concentration dependently with a maximal inhibitory effect at 5-10 microM. Despite the toxic effect at 20 microM, sulfonylureas showed an inhibitory effect. N-Acetylcysteine, superoxide dismutase, catalase, 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide and Mn(III) tetrakis(4-benzoic acid)porphyrin chloride inhibited the cytotoxicity of MPP+. Glibenclamide attenuated the nuclear damage, changes in the mitochondrial membrane permeability, caspase-3 activation and formation of reactive oxygen species due to MPP+ in PC12 cells. The results show that glibenclamide may reduce the MPP+ induced viability loss in PC12 cells by suppressing the changes in the mitochondrial membrane permeability, leading to the release of cytochrome c and subsequent activation of caspase-3, which are associated with the increased reactive oxygen species formation and depletion of GSH. PMID- 16310185 TI - Increased EMMPRIN (CD 147) expression during oral carcinogenesis. AB - Gene expression profiling of oral premalignant (OPM) cells and normal oral epithelial (NOR) cells showed that EMMPRIN expression was markedly upregulated in OPM cells compared to NOR cells. We used an oral squamous cell carcinoma (OSCC) progression model composed of cell lines, organotypic cultures and tissue specimens to characterize EMMPRIN expression patterns by microarray analysis, qRT PCR, Western blotting and immunohistochemistry. EMMPRIN levels are elevated in OPM and primary and metastatic OSCC cells as compared to NOR. EMMPRIN was detected as high and low glycosylated forms in the OPM and OSCC cellular extracts and was released in the media by OSCC cells but not by OPM cells. EMMPRIN expression in an organotypic culture model of normal and OPM mucosae mirrored the expression patterns in the respective tissues in vivo. EMMPRIN expression was limited to basal cells of normal, benign hyperkeratotic and inflammatory (lichen planus) oral mucosa. EMMPRIN expression is increased in dysplastic leukoplakias spreading to more superficial layers, and its expression levels correlated significantly with the degree of dysplasia. Primary and metastatic OSCC showed strong cell surface expression of EMMPRIN. These results suggest that EMMPRIN overexpression occurs at a very early stage of oral carcinogenesis and plays a contributing role in OSCC tumorigenesis. PMID- 16310186 TI - Vault mobility depends in part on microtubules and vaults can be recruited to the nuclear envelope. AB - Vaults are ribonucleoproteins that may function in intracellular transport processes. We investigated the intracellular distribution and dynamics of vaults in non-small cell lung cancer cells in which vaults are labeled with the green fluorescent protein. Immunofluorescence experiments showed that vaults are dispersed throughout the cytoplasm; a small fraction is found in close proximity to microtubules. Immunoprecipitation experiments corroborated these results showing co-precipitation of MVP and beta-tubulin. Using quantitative fluorescence recovery after photobleaching (FRAP), we demonstrated that vault mobility over longer distances in part depends on intact microtubules; vaults moving slower when microtubules are depolymerized by nocodazole. Biochemical fractionation indicated a small fraction of MVP associated with the nucleus, however, no GFP tagged vaults could be observed inside the nucleus. We observed an accumulation of vaults at the nuclear envelope upon treatment of cells with the protein synthesis inhibitor cycloheximide. Analysis of nucleo-cytoplasmic transport using a fluorescent substrate containing a classical NLS and NES expressed in MVP+/+ and MVP-/- mouse embryonic fibroblasts indicated no differences in nuclear import/export kinetics, suggesting no role for vaults in these processes. We hypothesize that a subset of vaults moves directionally via microtubules, possibly towards the nucleus. PMID- 16310187 TI - Androgen receptor and NFkB expression in human normal and glaucomatous optic nerve head astrocytes in vitro and in experimental glaucoma. AB - For several decades, clinical and experimental observations suggested a relationship between steroids and glaucoma; however, the possibility that androgens are also involved in the glaucomatous changes in the optic nerve heads (ONH) has not been explored. Our previous findings that glaucomatous ONH astrocytes synthesize androgen-metabolising enzymes and overproduce a neuroactive androgen, 5alpha-androstane-3alpha, 17beta-diol (3alpha-diol) led us to propose that ONH astrocytes are androgen target cells. Androgens modulate different cellular processes through androgen receptor (AR). NFkB is a transcription factor that positively regulates AR transcription. Here, we analysed AR and NFkB expression in normal and glaucomatous ONH astrocytes in vitro, and in vivo in a monkey model of experimental glaucoma (ExpG) by quantitative real time RT-PCR, Western blotting and immunohistochemistry. We demonstrated that in vitro human glaucomatous ONH astrocytes express AR mRNA and protein at higher levels than normal astrocytes and that in vivo ONH astrocytes from eyes with ExpG showed increased nuclear and cytoplasmic AR immunostaining compared to control eyes. In the retina, retinal ganglion cells (RGC) demonstrated cytoplasmic staining both in control and in ExpG eyes. NFkB mRNA expression was higher in glaucomatous ONH astrocytes than in normal and more nuclear NFkB protein was detected in glaucomatous ONH astrocytes. In vivo immunopositive NFkB nuclear staining of ONH astrocytes in ONH and in RGC in retina was detected both in control and in ExpG eyes. We conclude that in addition to our published data, increase of AR and NFkB expression in glaucomatous ONH astrocytes provides strong evidence that androgens play a significant role in the pathophysiology of glaucoma. PMID- 16310188 TI - Focus on molecules: X-linked Retinitis Pigmentosa 2 protein, RP2. PMID- 16310189 TI - Mouse model of sustained elevation in intraocular pressure produced by episcleral vein occlusion. AB - We have developed an inducible mouse model of glaucoma based on episcleral vein cauterization (EVC). Intraocular pressure (IOP) elevation in adult mice was produced by cauterizing three episcleral veins. Serial IOP measurements were done by induction-impact tonometry. IOP was significantly elevated by 104+/-20% in 20 out of 23 mice (87%) within the first day after EVC, and remained elevated for 4 weeks, with mean IOP 94% higher in EVC-treated vs. contralateral control eyes. Aqueous outflow blockade was verified from the IOP response to pulsed fluid infusions into the anterior chamber. Retinal ganglion cell (RGC) loss, determined by retrograde labelling using Fluoro-Gold applied to the superior colliculous, was approximately 20% at 2 weeks after EVC. We conclude that episcleral vein occlusion in mice produces significant and sustained elevation in IOP associated with increased outflow resistance and RGC loss, and thus may be useful to model glaucoma in genetically modified and drug-treated mice. PMID- 16310190 TI - Critical role of the Rho-kinase pathway in TGF-beta2-dependent collagen gel contraction by retinal pigment epithelial cells. AB - Retinal pigment epithelial cells (RPEs) are thought to be one of the main components of fibrous membrane observed in eyes with proliferative vitreo retinopathy. We investigated the signalling mechanisms of TGF-beta2-dependent collagen gel contraction by RPEs. An in vitro type I collagen gel contraction assay was performed to evaluate the effect of TGF-beta2 on gel contraction. The expression of alpha-smooth muscle actin (alpha-SMA) and the phosphorylation state of myosin light chain (MLC) were analyzed by Western blotting. The involvement of protein kinases such as p44/42 mitogen-activated protein kinase (MAPK), protein kinase C (PKC), p38 MAPK and phosphatidylinositol-3 kinase was investigated. The contribution of Rho-kinase and/or MLC-kinase was also evaluated using respective kinase inhibitors (Y27632, hydroxyfasudil and ML7). Additionally, RPEs were immunostained to examine whether the expression of alpha-SMA detected in our western blotting correlated to the stress fiber formation within the cells. TGF beta2 caused time (0-5 days)-and dose (0 10 ng ml(-1))-dependent gel contraction associated with overexpression of alpha-SMA and phosphorylation of MLC (p < 0.01, respectively). PKC inhibitor (GF109203X, 5 microM) and p38 MAPK inhibitor (SB203580, 10 microM) significantly attenuated TGF-beta2-elicited gel contraction via partial downregulation of both alpha-SMA expression and MLC phosphorylation (p < 0.01, respectively). The gel contraction was prominently inhibited in the presence of Y27632 (10 microM) or hydroxyfasudil (10 microM) with strong suppression of MLC phosphorylation but had no significant effect on alpha-SMA expression. Treatment with ML7, in contrast, resulted in a marginal inhibition of MLC phosphorylation and gel contraction. Finally, pretreatment of the cells with Y27632 or hydroxyfasudil prevented the formation of stress fiber within the cells. These results indicate that TGF-beta2-dependent myofibroblastic transdifferentiation and MLC phosphorylation by RPEs involve both PKC and p38 MAPK pathways at least in part. Myofibroblastic transdifferentiation of RPEs appears to be independent of the Rho-kinase pathway, and the presence of alpha SMA does not necessarily reflect the contractile potential of a cell. While Rho kinase inhibitors are incapable of preventing myofibroblastic transdifferentiation itself, this pathway could be one of the critical targets of cell-mediated contraction of the tissue containing fibrillar collagens by transdifferentiated RPEs. PMID- 16310191 TI - Characterization of free-floating spheres from human trabecular meshwork (HTM) cell culture in vitro. AB - It has been observed in several tissues that direct isolation of cells in serum free media and on nonadhesive substrates results in the formation of spherical clusters of cells known as free-floating spheres. Such free-floating spheres have been hypothesized to contain undifferentiated multipotent progenitor cells. Our goal was to isolate and characterize such free-floating spheres from HTM cell primary cultures. For this purpose, HTM cells were incubated in serum-free media and on a nonadhesive substrate. Individual free-floating spheres generated in these conditions were isolated in 96-well plates, and their proliferative capacity was evaluated by monitoring their size increase over time. The expression of the TM markers, MGP and CHI3L1, was examined using recombinant adenoviruses containing the respective promoters. Morphology of the free-floating spheres was analysed in semithin sections, and the gene expression profile was obtained using Human Genome U133 Plus 2.0 Affymetrix microarrays. HTM cells incubated in serum-free media and on nonadhesive substrate generated free floating spheres that could be grown for more than 3 months. Addition of serum to the culture media promoted the attachment of the spheres to the substrate, migration of cells from the spheres, and differentiation into cells phenotypically similar to normal TM cells. Gene profiling analysis demonstrated strong similarities between the gene expression profiles of the spheres and HTM cell monolayers. Both infection with the recombinant adenoviruses and gene array analysis demonstrated the expression of CHI3L1 and MGP, indicating that free floating spheres likely originate from HTM cells. Gene array analysis also showed expression of the marker for neural precursor cells nestin, as well as leukemia inhibitory factor, a gene involved in the maintenance of the undifferentiated state of progenitor cells. Analysis of semithin sections indicated that these TM free-floating spheres were highly dynamic structures demonstrating a distinct radial gradient of cell proliferation, survival, and apoptosis. Extensive up- and down-regulation of gene expression was associated with the processes of sphere attachment and cell migration after the addition of serum. These results suggest that HTM primary cultures might contain relatively undifferentiated or progenitor cells. The availability of TM progenitor cell cultures could constitute a useful tool to investigate cell therapy approaches targeting the TM in glaucoma. PMID- 16310192 TI - Literature review of current techniques for the insertion of distal screws into intramedullary locking nails. AB - The procedure for distal interlocking of intramedullary nails can be difficult and time consuming. Following nail insertion, the problems associated with the location of the distal holes and correct screw placements are well known. Numerous techniques and devices have been proposed to aid distal targeting, in attempts to overcome some of the associated problems. The development of the techniques and devices continues. A review of the literature is therefore timely, in order to provide awareness of the current situation. An overview is presented of the various distal targeting methods reported in the literature. PMID- 16310193 TI - Fractures of the femoral neck in children: long-term follow-up in 62 hip fractures. PMID- 16310194 TI - Re: Necrosis of the femoral head after fixation of trochanteric fractures with Gamma Locking Nail. A cause of late mechanical failure. PMID- 16310195 TI - Relationship between perceived empowerment care and quality of life among elderly residents within nursing homes in Taiwan: a questionnaire survey. AB - BACKGROUND: It is essential for nurses to have a solid understanding of what may influence the quality of life (QOL) among elderly residents within nursing homes. Although many factors have been demonstrated to be related to the QOL among elderly people, the relationship between perceived empowerment care and QOL among elderly residents within nursing homes has not been investigated thoroughly. OBJECTIVES: The aim of this study is to explore how demographic characteristics, perceived empowerment care and functional status affect perceived QOL among the elderly residents who reside in nursing homes. DESIGN: This study employed a cross-sectional design. SETTINGS: Eight nursing homes throughout southern Taiwan were recruited into this study. PARTICIPANTS: One hundred and two residents aged 65 years or older residing in nursing homes for at least 6 months, who did not have moderate to severe obstacles in cognitive ability, were recruited as participants. METHODS: A questionnaire including demographic data, a physical function scale, an activity of daily living (ADL) scale, a patient empowerment scale, and a QOL index was used in this study. RESULTS: The results showed a medium rating level of QOL among elderly residents. Elderly residents did not feel excessively disempowered care. Perceived empowering care, ADLs and marital status were significant predictors of QOL, which explained 45.3% of the total amount of variance in QOL. Perceived empowering care was the most important predictor of QOL. CONCLUSIONS: Nurses should pay attention to the effect of empowering care on QOL of elderly residents within nursing homes. Empowering care should be considered as a strategy of nursing care in nursing homes. PMID- 16310196 TI - Interactions between family members and staff in intensive care units--an observation and interview study. AB - BACKGROUND: Research about interactions between family members and staff are sparse, although family members' needs and experiences in intensive care units are well researched areas. AIM: The aim was to describe and interpret interactions between family members and staff in intensive care units. METHODS: Interviews (n=24) with family members and repeated observations of interactions of family members and staff in intensive care units were performed and analysed together by means of content analysis. RESULTS: The initial interactions between staff and family members had a substantial effect on family members and influenced their further interactions with the staff. Two kinds of interactions were revealed; mutual understanding and mutual misunderstanding. Family members, who understood the explicit information and the implicit messages were open in communication with the staff, adjusted well to the system, were acknowledged by the staff and sometimes consoled. Family members, who had difficulties understanding information and implicit messages drew back from communication with staff, did not adjust to the system and were sometimes insulted by the staff. CONCLUSIONS: Unambiguous information from the staff is important for developing interactions of mutual understanding. The results may be a starting point for intensive care unit staff to reflect on how all family members are initially met and further informed and treated. Further research of family members' experiences of interactions with staff in a longitudinal perspective and the influence of critical illness on families are needed. PMID- 16310197 TI - Polyphenols synergistically inhibit oxidative stress in subjects given red and white wine. AB - Aim of this study was to analyse the relationship between the plasma levels of polyphenols and the antioxidant activity of red and white wine. Twenty healthy subjects (HS) were randomly allocated to drink 300 ml of red (n = 10) or white n = 10 wine for 15 days. Ten HS who refrained from any alcohol beverage for 15 days were used as control. Urinary PGF-2alpha-III, a marker of oxidative stress and plasma levels of polyphenols were measured. Urinary PGF-2alpha-III significantly fell in subjects taking wine with a higher percentage decrease in subjects given red wine (-38.5 +/- 6%, p < 0.001) than in those given white wine (-23.1 +/- 6%). Subjects taking red wine had higher plasma polyphenols than those taking white wine (1.9 +/- 0.6 microM versus 1.5 +/- 0.33 microM, p < 0.001). Plasma polyphenols were inversely correlated with urinary PGF2alpha (r = 0.77, p < 0.001). No changes of urinary isoprostanes were observed in subjects who refrained from wine intake. In vitro study demonstrated that only a mixture of polyphenols, all in a range corresponding to that found in human circulation, inhibited LDL oxidation and PKC-mediated NADPH oxidase activation. Such inhibitory effects were more marked using the concentrations of polyphenols detected in human circulation after red wine intake. This study shows that red wine is more antioxidant than white wine in virtue of its higher content of polyphenols, an effect that may be dependent upon a synergism among polyphenols. PMID- 16310198 TI - LDL-cholesterol lowering or HDL-cholesterol raising for cardiovascular prevention. A lesson from cholesterol turnover studies and others. AB - A number of reports have indicated that both lowering low density lipoprotein (LDL)-cholesterol and raising high density lipoprotein (HDL)-cholesterol can result in significant cardiovascular benefit, both in terms of reduction of events and also, to a variable extent, of atheromatous lesions. LDL and HDL have opposite roles in body cholesterol regulation and, in theory, both reduced deposition (LDL reduction) and increased removal (raised HDL) can improve vascular disease. A number of reports over the last 30 years have attempted to quantitate with cholesterol balance/turnover studies, the correlations between LDL and HDL levels and body cholesterol pool sizes. More recently, these studies have evaluated the effects of LDL or HDL changes on cholesterol elimination. Data have, at times, been fully consistent with theoretical expectations, whereas at others they have not. Evaluation of these, at times, historical data provides, however, an important clue to the understanding of current results with different medications for the management of lipoprotein disorders. PMID- 16310199 TI - Determination of flavonoids in cultivated sugarcane leaves, bagasse, juice and in transgenic sugarcane by liquid chromatography-UV detection. AB - A high-performance liquid chromatography (HPLC) method with photo-diode array (DAD) detection was developed to separate and quantify flavonoids in sugarcane leaves and bagasse (= the crushed sugarcane refuse from juice extraction), and in sugarcane juice. Sugarcane flavonoids consist of a complex mixture of aglycones and glycosides (including flavonolignan glycosides), and the HPLC-UV method herein proposed is suitable for their quantification as total flavonoids. This method was applied to analyze samples of cultivated sugarcane, commercial juice and transgenic sugarcane leaves. Sugarcane leaves proved a promising source of flavonoids: an average of 1.10 mg of total flavonoids/g plant material was found in fresh leaves. Moreover, the flavonoid content of sugarcane juice (0.6 mg/mL) is comparable to other food sources of flavonoids previously reported. Transgenic sugarcane leaves ("Bowman-Birk" and "Kunitz") were compared with non-modified ("control") plant samples using the proposed HPLC-UV method, which indicated that the content of total flavonoids in transgenic plants is different from that in non-modified sugarcane. PMID- 16310200 TI - Determination of biogenic amines by capillary zone electrophoresis with conductometric detection. AB - A capillary zone electrophoresis (CZE) method with conductometric detection of biogenic amines (cadaverine, putrescine, agmatine, histamine, tryptamine and tyramine) is described. The optimised background electrolyte was the following: 15 mM histidine + 5 mM adipic acid + 1.5 mM sulphuric acid + 0.1 mM ethylenediaminotetraacetic acid + 0.1% hydroxyethylcellulose + 50% methanol. A clear separation of six biogenic amines from other components of acidic sample extract was achieved within 10 min. Method characteristics, i.e., linearity (0 100 micromol/ml), accuracy (recovery 86-107%), intra-assay repeatability (2-4%), and detection limit (2-5 micromol/l) were evaluated. Low laboriousness, sufficient sensitivity, speed of analysis, and low running cost are important attributes of this method. The developed method was successfully applied on the determination of biogenic amines in selected food samples. PMID- 16310201 TI - Effect of gravitational force on type-J counter-current chromatography by mathematical analysis. AB - The gravitational force exerts an important effect on the counter-current chromatography (CCC) and has been used to retain the stationary phase and improve the efficiency in the various CCC separations. This paper deals with the effect of gravitational force on type-J CCC, one of the most popular CCC methods by the mathematical analysis for the first time. The theoretical analyses reveal a close relationship between the effect of gravitational force, the placed orientation of type-J CCC apparatus, and the revolutional speed. Two placed orientations are more suitable for preparative CCC separation: one is common horizontal, and the other upright. Thus, there are three optimum conditions for type-J CCC separation, the first using horizontal apparatus at very low revolutional speed around 10rpm in the gravitational filed, the second by use of upright apparatus at moderate revolutional speed around 100rpm in the three-dimensional complex force field composed of centrifugal and orthogonal gravitational force, and the third employing the apparatus placed at any orientation including horizontal and upright CCC apparatus at high revolutional speed above 300rpm in the centrifugal force field. Therefore, we should consider the effect of gravitation force on CCC at the selection of proper operational conditions according to properties of two immiscible phases, the desired revolutional speed and the parameters of apparatus in order to utilize efficiently the effect of gravitational force. These analyses and results will benefit to improve the efficiency of CCC separation, especially large-scale industrial preparation. PMID- 16310202 TI - Using of S-(-)-2-hydroxymethyl-1,1-dimethylpyrrolidinium tetrafluoroborate as additive to the background electrolyte in capillary electrophoresis. AB - We synthesised and used new type of quaternary ammonium salt [S-(-)-2 hydroxymethyl-1,1-dimethylpyrrolidinium tetrafluoroborate] as effective additive to acidic background electrolytes. We used this quaternary ammonium salt as effective agent for capillary zone electrophoresis separation of model mixture of five tricyclic antidepressants (amitriptyline, nortriptyline, imipramine, desipramine and clomipramine) as model analytes. We observed that addition of S-( )-2-hydroxymethyl-1,1-dimethylpyrrolidinium tetrafluoroborate ([HMDP](+) [BF(4)]( )) to acidic background electrolytes leads to suppression of magnitude of electroosmotic flow (EOF) and gradually change the direction of the EOF. Baseline separation of five TAs was achieved by using of 91.1 mmol L(-1) (20 gL(-1)) of [HMDP](+) [BF(4)](-) in 25 mmol L(-1) sodium phosphate pH 2.5, where electroosmotic mobility was -11.3 x 10(-9) m(2) V(-1) s(-1). We achieved baseline separation of five TAs with using of [HMDP](+) [BF(4)](-) as water solution too. We observed that [HMDP](+) [BF(4)](-) can be used as buffer additive, which offers relatively smaller anodic electroosmotic flow instead of cationic surfactants that are mostly used for genarating of anodic electroosmotic flow in capillary electrophoresis. PMID- 16310203 TI - Accurate measurements of thermodynamic properties of solutes in ionic liquids using inverse gas chromatography. AB - Activity coefficients at infinite dilution of 29 organic compounds in two room temperature ionic liquids were determined using inverse gas chromatography. The measurements were carried out at different temperatures between 323.15 and 343.15K. To establish the influence of concurrent retention mechanisms on the accuracy of activity coefficients at infinite dilution for 1-butyl-3 methylimidazolium octyl sulfate and 1-ethyl-3-methylimidazolium tosylate, phase loading studies of the net retention volume per gram of packing as a function of the percent phase loading were used. It is shown that most of the solutes are retained largely by partition with a small contribution from adsorption on 1 butyl-3-methylimidazolium octyl sulfate and that the n-alkanes are retained predominantly by interfacial adsorption on 1-ethyl-3-methylimidazolium tosylate. PMID- 16310204 TI - Optimization of operating conditions for the determination of perchlorate in biological samples using preconcentration/preelution ion chromatography. AB - Perchlorate originates as a contaminant in the environment from the use of salts in the manufacture of solid rocket fuels and munitions. Monitoring potential perchlorate contamination in the environment is of interest, however, very few analytical methods have been developed for perchlorate determination in biological samples. Analysis of complex samples by ion chromatography is complicated by matrix components that can interfere with perchlorate determination. However, a recently developed preconcentration/preelution (PC/PE) ion chromatography method has demonstrated the capability to analyze certain complex samples such as high salinity water, milk, and hydroponic fertilizers. The ability of this method to reduce sample background and lower detection limits in ion chromatography for various biological samples was evaluated in this study. The PC/PE method was applicable to the analysis of kidneys, livers, zebrafish, quail eggs, lettuce, and urine. Optimal operating conditions were determined for each matrix. Ranges of optimal wash volumes were shorter when 15 mM NaOH prewash solutions were used compared with 10mM and good recovery was achieved for most matrices with an injection period > or =60s. Prewash solution concentration did not appear to significantly affect matrix background. The PC/PE method was capable of reducing sample background when compared to EPA Method 314.0, which resulted in detection limits, with the exception of zebrafish and urine, that were two-fold lower than those achieved with EPA Method 314.0. PMID- 16310205 TI - Analysis of acrylamide in food samples by capillary zone electrophoresis. AB - Conditions for the determination of acrylamide (AA) after derivatisation with 2 mercaptobenzoic acid by capillary zone electrophoresis were established. A derivatisation reagent-acrylamide ratio of 35:1 was selected as optimum and the reagent excess was not removed as it did not affect the determination of acrylamide by CZE. The best separation was achieved using a 40 mM phosphate buffer at pH 8.0, working at 25 kV in un-coated fused silica capillaries. Linear calibration curves over the range studied (0.3-100 microg mL(-1)), the limit of detection (0.07 microg mL(-1)), and both run-to-run (RSD values of 5.8 and 2.2% for concentration at low and medium concentration levels, respectively) and day to-day precisions (up to 11.2 and 6.7% at low and medium concentration levels, respectively) were established. Finally, the applicability of the CZE proposed methodology was demonstrated by analyzing levels of acrylamide present in different foodstuff products such as home made french fries, breakfast cereals and biscuits. PMID- 16310206 TI - Cigarette filter as sorbent for on-line coupling of solid-phase extraction to high-performance liquid chromatography for determination of polycyclic aromatic hydrocarbons in water. AB - An on-line solid-phase extraction (SPE) protocol using the cigarette filter as sorbent coupled with high-performance liquid chromatography (HPLC) was developed for simultaneous determination of trace naphthalene (NAPH), phenanthrene (PHEN), anthracene (ANT), fluoranthene (FLU), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP), and benzo(ghi)perylene (BghiP) in water samples. To on-line interface solid-phase extraction to HPLC, a preconcentration column packed with the cigarette filter was used to replace a conventional sample loop on the injector valve of the HPLC for on-line solid phase extraction. The sample solution was loaded and the analytes were then preconcentrated onto the preconcentration column. The collected analytes were subsequently eluted with a mobile phase of methanol-water (95:5). HPLC with a photodiode array detector was used for their separation and detection. The detection limits (S/N = 3) for preconcentrating 42 mL of sample solution ranged from 0.9 to 58.6 ng L(-1) at a sample throughput of 2 samples h(-1). The enhancement factors were in the range of 409-1710. The developed method was applied to the determination of trace NAPH, PHEN, ANT, FLU, BbF, BkF, BaP and BghiP in local river water samples. The recoveries of PAHs spiked in real water samples ranged from 87 to 115%. The precisions for nine replicate measurements of a standard mixture (NAPH: 4.0 microg L(-1), PHEN: 0.40 microg L(-1), ANT: 0.40 microg L(-1), FLU: 2.0 microg L(-1), BbF: 1.6 microg L(-1), BkF: 2.0 microg L( 1), BaP: 2.0 microg L(-1), BghiP: 1.7 microg L(-1)) were in the range of 1.2 5.1%. PMID- 16310207 TI - Miniaturization of sampling for chemical reaction monitoring by capillary electrophoresis. AB - Purpose-made capillary electrophoresis set-ups for reaction kinetics monitoring featuring two automated injectors allowed the easy reduction of the needed reactant amount down to 500 microL. The first set-up is similar to the cross injector used frequently in lab-on-chip designs while the other uses falling droplets for sample/buffer delivery. The versatility of the system was demonstrated by the analysis of oxidation of C-vitamin by hydrogen peroxide. Pseudo first order reaction rates about 10(-3) s(-1) were measured with RSD = 1 3% in one experiment and RSD = 20% in interday/person experiment. Plate numbers were typically around 5000-20,000. PMID- 16310208 TI - Uniform Y2O3 coating on multi-component phosphor powders by modified polyol process. AB - We have developed a method to coat the phosphors comprising of multi-component systems to prevent them from aging effect. Y2O3 was selected as a protective material because of its superior ability to resist against mercury adsorption and oxidation and its good UV transparency at the wavelength used for phosphor activation. By dispersing the phosphor particles in the polyol medium that contains the yttrium precursor, heterogeneous nucleation and growth on the particle surface were induced. This allows us to obtain a thick and uniform coating of the phosphors with amorphous Y2O3 sols. The heat-treatment condition for Y3O3 sols was carefully controlled to keep the phosphors from surface oxidation and at the same time to assure the complete crystallization and burn out of organic impurity by XRD and TG/DTA. The surface morphology of the coated phosphors was examined by SEM as a function of reaction time and temperature and precursor concentration. PMID- 16310209 TI - Interfacial tension of ethylene and aqueous solution of sodium dodecyl sulfate (SDS) in or near hydrate formation region. AB - The interfacial tensions between ethylene and an aqueous solution of SDS were measured using the pendant-drop method at 274.2 and 278.2 K and in the pressure range from 0.1 to 3.1 MPa, including hydrate formation points. The concentrations of sodium dodecyl sulfate (SDS) aqueous solution were 0, 100, 300, 500, 600, 700, 800, 900, and 1000 ppm. The effects of pressure on the critical micelle concentration (CMC) and the surface excess concentration were studied. It was demonstrated that both the CMC and the saturated surface excess concentration decreased with the increase of pressure. PMID- 16310210 TI - Vapor-induced film formation from low-Tg particles for different solvent compositions. AB - The photon transmission method was used to study latex film formation from poly(vinyl acetate) (PVAc) particles induced by two different solvents. Films with the same latex content were prepared from PVAc particles and exposed to vapor of ethanol-water and acetone-water mixtures in various compositions. Transmitted photon intensities, Itr, from these films increased with increasing vapor exposure time. The increase in Itr is attributed to the increase in crossing density at the polymer-polymer junction. The Prager-Tirrell model was employed to obtain the back-and-forth frequency, nu, of the reptating polymer chain during film formation induced by solvent vapor. It was observed that the produced nu values increase as the solvent content is increased for both solvents. Abilities of both solvents to form films were interpreted with the solubility parameters of the solvents and the PVAc. PMID- 16310211 TI - Porous structure of natural and modified clinoptilolites. AB - The evaluation of the pore-size distribution (PSD) of natural and modified mesoporous zeolites, i.e., clinoptilolites is presented. We demonstrate the SEM results showing that the pores of fracture-type from 25-50 nm to 100 nm in size between clinoptilolite grains, as well as pores between crystal aggregates up to 500 nm in size are present in the studied material. The detailed distribution of pore sizes and tortuosity factor of the above-mentioned materials are determined from the adsorption-desorption isotherms of nitrogen measured volumetrically at 77 K. To obtain the reliable pore size distribution (PSD) of the above-mentioned materials both adsorption and desorption branches of the experimental hysteresis loop are described simultaneously by recently developed corrugated pore structure model (CPSM) of Androutsopoulos and Salmas. Evaluated pore size distributions are characterized by well-defined smooth peaks placed in the region of the mesoporosity. Moreover, the mean pore diameter calculated from the classical static measurement of nitrogen adsorption at 77 K correspond very well to the pore diameters from SEM, showing the applicability of the CPSM for characterization of the porosity of natural zeolites. We conclude that classical static adsorption measurements combined with the proper modeling of the capillary condensation/evaporation phenomena are a powerful method which can be applied for pore structure characterization of natural and modified clinoptilolites. PMID- 16310212 TI - Biological stoichiometry of growth in Drosophila melanogaster. AB - We examined the relationship between growth rate, C:N:P stoichiometry, and nucleic acid content in Drosophila melanogaster. The "Growth Rate Hypothesis" predicts that N and P contents per unit body mass will be high during ontogenetic stages characterized by rapid growth, reflecting the large requirement for P-rich ribosomal RNA during these periods. The ratio of RNA:DNA also is predicted to change with changes in growth rate. Growth is rapid in early D. melanogaster larvae, slowing considerably just prior to pupation. As predicted, a positive relationship was found between growth rate and N and P content, but not C. Thus, body C:P and N:P ratios declined with increasing growth rate. The relationship between RNA content and growth rate also was positive. Additionally, the fraction of total body P contributed by ribosomal RNA increased with increasing growth rate. PMID- 16310213 TI - The Malpighian tubule: rapid insights from post-genomic biology. AB - Good osmoregulation is critical to the success of insects, and the Malpighian tubules play a key role in osmoregulation. Recently, the application of genetics and genomics to the Drosophila tubule has revealed far more extensive roles than ion and water transport. Microarray analysis shows that organic solute transporters dominate the tubule transcriptome. The tubule thus has the capability to excrete actively the broadest range of organic solutes and xenobiotics. Such transporters can produce unexpected, emergent roles for the whole tissue; e.g. the tubule is highly resistant to ouabain not because the Na+, K+ ATPase is unimportant, but because it co-localises with a potent alkaloid excretory mechanism. Reinforcing this role in excretion, the tubule expresses very high levels of a particular cytochrome P450s, glutathione-S-transferases and alcohol dehydrogenases which suggest that the tubule plays a major role in metabolism and detoxification of both endogenous solutes and xenobiotics, such as insecticides. Additionally, the tubule plays a significant role in immunity; tubules are capable of sensing bacterial challenge, and mounting an effective killing response by secretion of antimicrobial peptides, entirely independent of the fat body, the canonical immune tissue. The tubule has also proved to be a good model for some human renal disease, and to act as an organotypic 'testbed' for mammalian genes. The tubule can thus bask in a greatly enhanced reputation as a key tissue for an unexpectedly wide range of functions in the insect. PMID- 16310214 TI - Characterization of a novel satellite virus and a strain of Himetobi P virus (Dicistroviridae) from the brown planthopper, Nilaparvata lugens. AB - Two distinct spherical virus-like particles were purified from the brown planthopper, Nilaparvata lugens. One was a geographical isolate of Himetobi P virus (Cripavirus, Dicistroviridae). The other was 30 nm in diameter and contained positive-stranded RNA. The RNA was 1647 nucleotides in length and encoded only its own capsid protein, indicating that this particle is a satellite virus. Transmission tests showed that the satellite was transmitted vertically; however, its helper virus was unknown. We named this satellite Nilaparvata lugens commensal X virus (NLCXV). PMID- 16310215 TI - The ubiquitin domain superfold: structure-based sequence alignments and characterization of binding epitopes. AB - Ubiquitin-like domains are present, apart from ubiquitin-like proteins themselves, in many multidomain proteins involved in different signal transduction processes. The sequence conservation for all ubiquitin superfold family members is rather poor, even between subfamily members, leading to mistakes in sequence alignments using conventional sequence alignment methods. However, a correct alignment is essential, especially for in silico methods that predict binding partners on the basis of sequence and structure. In this study, using 3D-structural information we have generated and manually corrected sequence alignments for proteins of the five ubiquitin superfold subfamilies. On the basis of this alignment, we suggest domains for which structural information will be useful to allow homology modelling. In addition, we have analysed the energetic and electrostatic properties of ubiquitin-like domains in complex with various functional binding proteins using the protein design algorithm FoldX. On the basis of an in silico alanine-scanning mutagenesis, we provide a detailed binding epitope mapping of the hotspots of the ubiquitin domain fold, involved in the interaction with different domains and proteins. Finally, we provide a consensus fingerprint sequence that identifies all sequences described to belong to the ubiquitin superfold family. It is possible that the method that we describe may be applied to other domain families sharing a similar fold but having low levels of sequence homology. PMID- 16310216 TI - Structural basis of syndecan-4 phosphorylation as a molecular switch to regulate signaling. AB - The syndecan transmembrane proteoglycans are involved in the organization of the actin cytoskeleton and have important roles as cell surface receptors during cell matrix interactions. We have shown that the syndecan-4 cytoplasmic domain (4L) forms oligomeric complexes that bind to and stimulate PKCalpha activity in the presence of PtdIns(4,5)P2, emphasizing the importance of multimerization in the regulation of PKCalpha activation. Oligomerization of the cytoplasmic domain of syndecan-4 is regulated either positively by PtdIns(4,5)P2 or negatively by phosphorylation of serine 183. Phosphorylation results in reduced PKCalpha activity by inhibiting PtdIns(4,5)P2-dependent oligomerization of the syndecan-4 cytoplasmic domain. Data from NMR and gel-filtration chromatography show that the phosphorylated cytoplasmic domain (p-4L) exists as a dimer, similar to 4L, but not as higher-order oligomers. NMR analysis showed that the overall conformation of p-4L is a compact intertwined dimer with an unusually symmetric clamp shape, and its molecular surface is mostly positively charged. The two parallel strands form a cavity in the center of the dimeric twist. An especially marked effect of phosphorylation of the syndecan-4 cytoplasmic domain is a dramatic conformational change near the C2 region that ablates an interaction site with the PDZ domain of syntenin. Wound healing studies further suggest that syndecan-4 phosphorylation might influence cell migration behavior. We conclude that the phosphorylation (Ser183) of syndecan-4 can play a critical role as a molecular switch to regulate its functions through conformational change. PMID- 16310217 TI - The major outer membrane protein of Fusobacterium nucleatum (FomA) folds and inserts into lipid bilayers via parallel folding pathways. AB - Membrane protein insertion and folding was studied for the major outer membrane protein of Fusobacterium nucleatum (FomA), which is a voltage-dependent general diffusion porin. The transmembrane domain of FomA forms a beta-barrel that is predicted to consist of 14 beta-strands. Here, unfolded FomA is shown to insert and fold spontaneously and quantitatively into phospholipid bilayers upon dilution of the denaturant urea, which was shown previously only for outer membrane protein A (OmpA) of Escherichia coli. Folding of FomA is demonstrated by circular dichroism and fluorescence spectroscopy, by SDS-polyacrylamide gel electrophoresis, and by single-channel recordings. Refolded FomA had a single channel conductance of 1.1 nS at 1 M KCl, in agreement with the conductance of FomA isolated from membranes in native form. In contrast to OmpA, which forms a smaller eight-stranded beta-barrel domain, folding kinetics of the larger FomA were slower and provided evidence for parallel folding pathways of FomA into lipid bilayers. Two pathways were observed independent of membrane thickness with two different lipid bilayers, which were either composed of dicapryl phosphatidylcholine or dioleoyl phosphatidylcholine. This is the first observation of parallel membrane insertion and folding pathways of a beta-barrel membrane protein from an unfolded state in urea into lipid bilayers. The kinetics of both folding pathways depended on the chain length of the lipid and on temperature with estimated activation energies of 19 kJ/mol (dicapryl phosphatidylcholine) and 70 kJ/mol (dioleoyl phosphatidylcholine) for the faster pathways. PMID- 16310218 TI - Absorption by a moving spherical organelle in a heterogeneous cytoplasm: implications for the role of trafficking in a symplast. AB - An organelle which absorbs (or secretes) a particular factor will find its mass transfer rate diffusion-limited if it is stationary with respect to its ambient cytoplasm; but organellar motion will raise that limit as a non-decreasing function of the Peclet number P. It is shown analytically that (i) no Whitehead paradox need be encountered in the creeping flow regime and (ii) the flux of the factor will be an even function of the Peclet number, P. By a novel analytic solution method, the flux is shown numerically to increase as P2 for P < or = 1. For P > or = 10, a quasi-planar approximating geometry yields analytically a flux which increases as P1/3. These two solutions overlap smoothly in the range 1 < or = P > or = 10. For P approximately 1, convection should increase the mass flux by roughly 100%. PMID- 16310219 TI - Variance component analysis of polymorphic metabolic systems. AB - The relationship between mechanistic allelic interaction in multi-gene systems and genetic contribution to population variance remains poorly understood. In order to address this problem, dynamic cellular processes must be reconciled with individual differences in a population. We suggest an approach to enable this for metabolic systems, whereby steady-state biomarker concentrations are calculated for individual systems carrying different alleles. As proof of principle, we simulated two versions of a three enzyme linear synthesis pathway, in a multi level framework from transcription to enzyme action. The first (Standard) model incorporated conventional kinetics, whilst an analogous model included negative feedback in the form of competitive inhibition (CI). Alleles were allowed to confer different transcription rates, and genetic components of variance in biomarker concentration calculated for populations of each system type. Initial simulations of high and low expression alleles revealed substantial genetic additivity and some dominance for both system architectures. For the Standard model population, each of the three genes contributed equally, whereas CI substantially altered the relative importance of individual genes. Epistasis was limited for both model populations, never rising above 5% in extensive parameter explorations. Subsequent simulations examined a wide range of allelic transcription rates, from almost null to three orders of magnitude above baseline. Again, for both model architectures, additive and dominance effects were most prevalent, but epistasis increased substantially as allelic effects approached null. We conclude that the nature of allelic contribution to variance is dependent upon both the magnitude of conferred effect and the structure of the system in which it is embedded, and relatively little on other system parameters. We believe that implementation of this approach holds promise of a better understanding of the genotype to phenotype transition, particularly in integrating small allelic effects into larger frameworks. PMID- 16310220 TI - Evolution of the activity of UGT1A1 throughout the development and adult life in a rat. AB - Biliary excretion is the main route of disposal of bilirubin and impaired excretion results in jaundice, a well recognisable symptom of liver disease. Conjugation of bilirubin in the liver is essential for its clearance. The glucuronidation of bilirubin is catalysed by the microsomal UDP glucuronosyltransferase UGT1A1. Patients with Crigler-Najjar syndrome type 1 and Gunn rats, mutant strain of the Wistar rats, bear an autosomal recessive disorder resulting in hyperbilirubinemia. The aim of this work is to add new data about activity of UGT1A1 during the perinatal period and adult life. The results showed that activity of UGT1A1 is detectable from day 22 of the gestation. After birth, activity of UGT1A1 gradually increases and reaches the levels of adult life. Furthermore, bilirubin azopigments have been separated and characterized by thin layer chromatography. We have found that concentration of samples by evaporation and ulterior storing at -20 degrees C seemed to be suitable for the maintenance of samples. PMID- 16310221 TI - Polysaccharide peptides from COV-1 strain of Coriolus versicolor induce hyperalgesia via inflammatory mediator release in the mouse. AB - Polysaccharide peptide (PSP), isolated from Coriolus versicolor COV-1, has been widely used as an adjunct to cancer chemotherapy and as an immuno-stimulator in China. In this study, the anti-nociceptive effects of PSP were investigated in two different pain models in the mouse. In the acetic acid-induced writhing model, initial studies showed that PSP decreased the number of acetic acid induced writhing by 92.9%, which, by definition, would constitute an analgesic effect. However, further studies showed that PSP itself induced a dose-dependent writhing response. Studies on inflammatory mediator release showed that PSP increased the release of prostaglandin E2, tumor necrosis factor-alpha, interleukin-1beta, and histamine in mouse peritoneal macrophages and mast cells both in vitro and in vivo. The role of inflammatory mediator release in PSP induced writhing was confirmed when diclofenac and dexamethasone decreased the number of writhing responses by 54% and 58.5%, respectively. Diphenhydramine totally inhibited the PSP-induced writhing. In the hot-plate test, PSP dose dependently shortened the hind paw withdrawal latency, indicative of a hyperalgesic effect. The hyperalgesic effect was reduced by pretreatment with the anti-inflammatory drugs. In conclusion, the PSP-induced hyperalgesia was related to activation of peritoneal resident cells and an increase in the release of inflammatory mediators. PMID- 16310222 TI - Strong electronic charge as an important factor for anticancer activity of chitooligosaccharides (COS). AB - Even though several studies report the importance of chitosan derivatives for their anticancer activity, no clear information is available to describe the relationship between their charge properties and observed activities. In this research, differently charged chitooligosaccharide (COS) derivatives were synthesized and their anticancer activities were studied using three cancer cell lines, HeLa, Hep3B and SW480. Neutral red and MTT cell viability studies revealed that, highly charged COS derivatives could significantly reduce cancer cell viability, regardless to the positive or negative charge. Further, fluorescence microscopic observations and DNA fragmentation studies confirmed that the anticancer effect of these highly charged COS derivatives were due to necrosis. However, the exact molecular mechanism for anticancer activity of strongly charged COS compared to their poorly charged counterparts is not clear. PMID- 16310223 TI - Acute changes of biventricular gene expression in volume and right ventricular pressure overload. AB - OBJECTIVE: We investigated the effects of acute volume and RV pressure overload on biventricular function and gene expression of BNP, pro-inflammatory cytokines (IL-6 and TNF-alpha), iNOS, growth factors (IGF-1, ppET-1), ACE and Ca2+-handling proteins (SERCA2a, phospholamban and calsequestrin). METHODS: Male Wistar rats (n=45) instrumented with pressure tip micromanometers in right (RV) and left ventricular (LV) cavities were assigned to one of three protocols: i) Acute RV pressure overload induced by pulmonary trunk banding in order to double RV peak systolic pressure, during 120 or 360 min; ii) acute volume overload induced by dextran40 infusion (5 ml/h), during 120 or 360 min; iii) Sham. RV and LV samples were collected for mRNA quantification. RESULTS: BNP upregulation was restricted to the overloaded ventricles. TNF-alpha, IL-6, ppET-1, SERCA2a and phospholamban gene activation was higher in volume than in pressure overload. IGF-1 overexpression was similar in both types of overload, but was limited to the RV. TNF-alpha and CSQ mRNA levels were increased in the non-overloaded LV after pulmonary trunk banding. No significant changes were detected in ACE or iNOS expression. RV end-diastolic pressures positively correlated with local expression of BNP, TNF-alpha, IL-6, IGF-1, ppET-1 and SERCA2a, while RV peak systolic pressures correlated only with local expression of IL-6, IGF-1 and ppET 1. CONCLUSIONS: Acute cardiac overload alters myocardial gene expression profile, distinctly in volume and pressure overload. These changes correlate more closely with diastolic than with systolic load. Nonetheless, gene activation is also present in the non-overloaded LV of selectively RV overloaded hearts. PMID- 16310224 TI - Resveratrol inhibits gentamicin-induced mesangial cell contraction. AB - Gentamicin is an aminoglycoside antibiotic that is very effective in treating different gram negative infections, however, one of its main side effects is nephrotoxicity. Gentamicin-induced decreases in glomerular filtration rate could be mediated by mesangial cell contraction. Resveratrol, a natural hydroxystilbene, has been identified to be a potent antioxidant with many biological activities including protection against kidney diseases. As we have previously demonstrated that gentamicin induced a reduction of planar surface area of cultured rat mesangial cells, and that resveratrol has a protective effect on gentamicin-induced nephrotoxicity in vivo, the aim of this study was to investigate the effect of resveratrol on gentamicin-induced mesangial cell contraction. This study demonstrates that the contractile effect of gentamicin on mesangial cells can be prevented by incubation with resveratrol at an optimal dose of 10 microM, as it blunted the gentamicin-induced reduction in planar cell surface area and the number of contracted cells. Besides, the preincubation with 10(-5)M diphenylene iodinium (DPI), an inhibitor of the NADP(H) oxidase, also blunted gentamicin-induced cell contraction. This preventive effect was higher when cells were incubated with both substances together. These results strongly suggest that the protective effect resveratrol against gentamicin-induced reduction in renal function in vivo could be mediated by inhibiting gentamicin induced mesangial cells contraction. PMID- 16310225 TI - Estrogenic effect of procymidone through activation of MAPK in MCF-7 breast carcinoma cell line. AB - Procymidone modifies sexual differentiation in vitro and induces estrogenic activity in primary cultured rainbow trout hepatocytes, as shown by an increase in the contents of vitellogenin and heat shock proteins. Since this dicarboximide fungicide is found in human tissues, it was considered of interest to investigate its ability to induce endocrine damage in the MCF-7 human cell line. The mechanism of this estrogenic action was also evaluated. Procymidone 100 microM stimulated cell growth from day 3 up to day 12 and raised the level of pS2 on day 3. Although procymidone does not bind the estrogen receptor (ER), the antiestrogen ICI 182780 inhibited its effect on cell growth and pS2 content, suggesting that the ER is involved indirectly in these effects. In exploring the mechanism of ER indirect activation we found that the antibody against c-Neu receptor (9G6) did not modify procymidone's effects on cell growth and pS2 expression. Thus, procymidone does not bind the c-Neu membrane receptor, excluding this indirect ER activation pathway. We also found that procymidone induced mitogen-activated protein kinase (MAPK) at 15 and 30 min, and that PD 98059, a MAPK (Erk1/2) inhibitor, prevented procymidone's effects on cell growth and pS2, indicating that MAPK activation is responsible for procymidone ER activation. The production of reactive oxygen species (ROS) with these times and elimination of the phenomenon by alpha-tocopherol (alpha-T), a ROS scavenger, is proof that oxygen free-radical production is at the basis of the MAPK activation by procymidone. PMID- 16310226 TI - Spatial distribution and ecotoxicity of petroleum hydrocarbons in sediments from the Galicia continental shelf (NW Spain) after the Prestige oil spill. AB - The distribution of aliphatic and aromatic hydrocarbons was determined in surface sediments collected at 36 stations along the Galicia continental shelf (NW Spain), following the Prestige oil spill. Sampling was performed in December 2002, just after the accident, and in February and September 2003. Concentrations of PAHs (summation operator 13 parent components) were in the range of 0.9-422 microg/kgdw, the highest values being close to coastal urban areas (e.g. Pontevedra and A Coruna), whereas in the stations of the area most heavily impacted by the spill (off Costa da Morte) concentrations were in the range of 14.8-89.6 microg/kgdw, with a certain predominance of alkylated compounds, which may suggest a mixture of petrogenic and pyrolytic sources. The detailed study of petrogenic molecular markers (e.g. steranes and triterpanes) showed the occurrence of an old (weathered) petrogenic chronic pollution in the shelf sediments but not of the Prestige oil, with the possible exception of few stations in the area of Costa da Morte. This was attributed to the heavy nature of the spilled oil that was barely dispersed in the water column and mainly stranded on the coast or sedimented in the form of oil patches. The addition of increasing amounts of fuel oil to a representative sediment sample showed that the molecular indices were indicative of the presence of the Prestige oil when the amount was above 1g/kg of sediment. The toxicity of selected samples (showing the higher PAH concentrations) was tested using the bivalve embryogenesis bioassay. Embryogenesis success reached high values in all cases (80-88%, with 86% in the control), indicating a lack of toxicity in the sediments and supporting the conclusion that the patchiness of the fuel eventually reaching the seafloor reduced its impact on the benthic communities of the Galician shelf. PMID- 16310227 TI - Among the twenty classical L-amino acids, only glutamate directly activates metabotropic glutamate receptors. AB - Under pathophysiological conditions, cellular amino acids can be profusely released from cells into the cerebral interstitial space. Because several class-C G protein coupled receptors (GPCRs) display a broad natural ligand spectrum, being sensitive to more than one endogenous ligand, we wondered whether the related metabotropic glutamate (mGlu) receptors could be modulated by various types of L-amino acids, allowing them to sense large increase in extracellular amino acid concentration. Here, the agonist, antagonist and allosteric effects of the twenty classical L-amino acids were evaluated on the eight mGlu receptor subtypes. We show that, in addition to glutamate (Glu), cysteine, aspartate and asparagine also lead to the activation of mGlu3, 4 and 5. Interestingly, our data demonstrate that the effect of these three amino acids did not result from a direct activation of the receptors, but from an indirect action involving Glu transporters/exchangers. These data first demonstrate that mGlu receptors, unlike other class-C GPCRs, display an extremely high selectivity towards one ligand. Moreover, our results also show that Glu transport systems allow mGlu receptors to sense large increase in the extracellular concentration of some amino acids. Such a system will certainly lead to a large increase in some mGlu receptor activity under pathological conditions, such as seizure, ischemia or other brain injuries. PMID- 16310228 TI - The human Ca2+-activated K+ channel, IK, can be blocked by the tricyclic antihistamine promethazine. AB - Phenothiazines can be used as psychopharmaceutical agents and are known to cause many side effects during treatment since they interfere with many different cellular systems. Recently, phenothiazines were reported to block Ca(2+) activated potassium channels of the SK type. Therefore we investigated their effect on the functionally related class of Ca(2+)-activated potassium channels of the IK type. The representative phenothiazine derivative promethazine (PTZ) blocked IK channels almost independently from the extracellular pH(o) with an IC(50) of 49 +/- 0.2 microM (pH(o) 7.4, n = 5) and 32 +/- 0.2 microM (pH(o) 6.2, n = 5) in whole cell experiments. The extracellularly applied membrane impermeable PTZ analogue methyl-promethazine (M-PTZ) had a strongly reduced blocking potency compared to PTZ. In contrast, intracellularly applied PTZ and M PTZ had the same blocking potency on IK channels in excised inside out patch clamp experiments (K(d) = 9.3 +/- 0.5 microM for PTZ, n = 7 and 6.7 +/- 0.4 microM for M-PTZ, n = 5). The voltage dependency of the PTZ and M-PTZ block was investigated in excised inside out patch clamp experiments at a concentration of 100 microM. For both compounds the block was more pronounced at positive membrane potentials. The steepness of the voltage dependency was found to be 70 +/- 10 mV (for PTZ) and 61 +/- 6 mV (for M-PTZ) indicating that both compounds sensed approximately 40% of the entire membrane spanning electrical field from the inside. We conclude that PTZ and M-PTZ bind to a side in IK channels, which is located within the electrical field and is accessible from the intracellular side. PMID- 16310229 TI - Stimulation of Orobanche ramosa seed germination by fusicoccin derivatives: a structure-activity relationship study. AB - A structure-activity relationship study was conducted assaying 25 natural analogues and derivatives of fusicoccin (FC), and cotylenol, the aglycone of cotylenins, for their ability to stimulate the seed germination of the parasitic species Orobanche ramosa. Some of the compounds tested proved to be highly active, being 8,9-isopropylidene of the corresponding FC aglycone and the dideacetyl derivative the most active FC derivatives. In both groups of glucosides and aglycones (including cotylenol), the most important structural feature to impart activity appears to be the presence of the primary hydroxy group at C-19. Furthermore, the functionalities and the conformation of the carbotricyclic ring proved to play a significant role. The dideacetyl derivative of FC, being easily and rapidly obtainable in high yield starting by FC, could be of interest for its practical application as a stimulant of Orobanche ramosa seed germination, inducing the "suicidal germination", an interesting approach for parasitic plant management. PMID- 16310230 TI - Cuticular waxes from potato (Solanum tuberosum) leaves. AB - The qualitative and quantitative compositions of leaf cuticular waxes from potato (Solanum tuberosum) varieties were studied. The principal components of the waxes were very long chain n-alkanes, 2-methylalkanes and 3-methylalkanes (3.1-4.6 microg cm(-2)), primary alcohols (0.3-0.7 microg cm(-2)), fatty acids (0.3-0.6 microg cm(-2)), and wax esters (0.1-0.4 microg cm(-2)). Methyl ketones, sterols, beta-amyrin, benzoic acid esters and fatty acid methyl, ethyl, isopropyl and phenylethyl esters were found for the first time in potato waxes. The qualitative composition of the waxes was quite similar but there were quantitative differences between the varieties studied. A new group of cuticular wax constituents consisting of free 2-alkanols with odd and even numbers of carbon atoms ranging from C25 to C30 was identified. PMID- 16310231 TI - Xanthone derivatives from Cratoxylum cochinchinense roots. AB - Two xanthones and two caged-prenylated xanthones, named cochinchinones A-D, respectively, and a synthetically known caged-prenylated xanthone, together with seven known compounds were isolated from the roots of Cratoxylum cochinchinense (Lour.) Blume. Their structures were assigned on the basis of analyses of spectroscopic data. Some of the compounds exhibited effective antioxidative properties. PMID- 16310232 TI - Lignan, phenolic and iridoid glycosides from Stereospermum cylindricum. AB - A lignan glycoside [(+)-cycloolivil 4'-O-beta-d-glucopyranoside], a phenolic glycoside [3,4-dimethoxyphenyl 1-O-beta-d-xylopyranosyl-(1-->6)-beta-d glucopyranoside] and a iridoid glycoside (stereospermoside) were isolated from the leaves and branches of Stereospermum cylindricum, together with (+) cycloolivil, (+)-cycloolivil 6-O-beta-d-glucopyranoside, (-)-olivil, (-)-olivil 4 O-beta-d-glucopyranoside, (-)-olivil 4'-O-beta-d-glucopyranoside, vanilloloside, decaffeoyl-verbascoside, isoverbascoside, 3,4,5-trimethoxyphenyl 1-O-beta-d xylopyranosyl-(1-->6)-beta-d-glucopyranoside, ajugol, verminoside, and specioside. The structure elucidations were based on spectroscopic evidence. PMID- 16310233 TI - Wound-induced green leaf volatiles cause the release of acetylated derivatives and a terpenoid in maize. AB - Green leaf volatiles (GLVs), generally occurring C6 alcohols, aldehydes and acetates from plants, play an important role in plant-plant communication. These compounds induce intact plants to produce jasmonic acid, and induce defense related gene expression and the release of volatile compounds. Here, we address wound-induced GLVs cause the release of acetylated derivatives and a terpenoid, (E)-4,8-dimethylnona-1,3,7-triene (DMNT) in intact maize, which may be a type of plant-plant interaction mediated by airborne GLVs. Upon exposure of intact maize seedlings to wound-induced GLVs, (Z)-3-hexenyl acetate was consistently the most abundant compound released. Exogenous application of individual alcohols and aldehydes mostly resulted in the release of corresponding acetate esters. C6 alcohols with a double bond between the second and third, or the third and fourth carbon atoms, C5- or C6-aldehydes, and (Z)-3-hexenyl acetate triggered the release of DMNT. When (Z)-3-hexenyl acetate and hexyl acetate were used to treat maize seedlings, they were recovered from the plants. These data demonstrated that: (1) apart from direct adsorption and re-release of acetate esters, absorption and conversion of exogenous alcohols and aldehydes into acetate esters occurred, and (2) DMNT was induced by a range of aldehydes and unsaturated alcohols. PMID- 16310234 TI - Potential cancer chemopreventive activity of simple isoquinolines, 1 benzylisoquinolines, and protoberberines. AB - Seventeen simple isoquinolines, 15 1-benzylisoquinolines, and 19 protoberberines were tested for their inhibitory activities against Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells. Among the tested alkaloids, the inhibitory activity of all 1 benzylisoquinolines and 11 protoberberines was higher than that of beta-carotene. The 1-benzylisoquinolines 19, 21, 22, 29, and 34 and protoberberines 41, 47-49, 51, 52, and 55 showed potent inhibitory effects on EBV-EA induction (96-100% inhibition at 1 x 10(3) mol ratio/TPA). These alkaloids were more active than the naturally occurring alkaloids, 23, 25, 33, 53, and 54. In addition, fifteen simple isoquinolines, eighteen 1-benzylisoquinolines and eight protoberberines were evaluated with respect to their ability to scavenge 1,1-diphenyl-2 picrylhydrazyl (DPPH) free radicals. Nine simple isoquinolines, ten 1 benzylisoquinolines, and four protoberberines were more potent than alpha tocopherol, and four 1-benzylisoquinolines, 20 and 28-30, exhibited potent activities (SC50 4.5-5.8 microM). Their activities were higher than the naturally occurring alkaloids, 23, 25, and 33. Therefore, some of the isoquinoline alkaloids indicating the high activity on both assays may be potentially valuable cancer chemopreventive agents. Structure-activity relationships are discussed for both tests. PMID- 16310235 TI - Evaluation of the antibody response to Anopheles salivary antigens as a potential marker of risk of malaria. AB - The evaluation of human immune responses to arthropod bites may be a useful marker of exposure to vector-borne diseases, with applications to malaria, the most serious parasitic infection in humans. The specific antibody (Ab) IgG response to saliva obtained from Anopheles gambiae mosquitoes was evaluated in young children from an area of seasonal malaria transmission in Senegal. Specific IgG was higher in children who developed clinical Plasmodium falciparum malaria within the 3 months that followed than in those who did not (P<0.05), and it increased significantly (P<0.0001) with the level of Anopheles exposure, as evaluated by conventional entomological methods. These results suggest that evaluation of antisalivary Ab responses could be a useful approach for identifying a marker for the risk of malaria transmission. PMID- 16310236 TI - Pool screen PCR for estimating the prevalence of Leishmania infantum infection in sandflies (Diptera: Nematocera, Phlebotomidae). AB - Prevalence studies of infection in the sandfly vector can be used as an indicator of a change in the intensity of Leishmania transmission. However, these studies are difficult to carry out as prevalence in the vector is usually low and its estimation requires a large number of sandflies to be dissected. Our objective was to establish whether a L. infantum-specific PCR-ELISA applied to pools of female sandflies and a previously described algorithm could be useful tools to study the prevalence of infection by this parasite in natural vector populations. We collected sandflies from six collection points in two stable foci of leishmaniasis in southern (N=3) and north-eastern (N=3) Spain, following standard procedures. A fraction of the collected females was dissected and morphologically identified. Another fraction was used for pool screening. In total, 127 pools of 30 females (3810 specimens) were studied by PCR-ELISA and 1764 specimens were individually dissected. The prevalence of infection determined by dissection does not differ from that determined by pool screen PCR. The results suggest that pool screen PCR can be of practical use in the epidemiological surveillance of leishmaniasis in European countries of the western Mediterranean basin, associated with control interventions or global change. PMID- 16310237 TI - Venom production in long-term primary culture of secretory cells of the Bothrops jararaca venom gland. AB - There is an increasing interest of obtaining venom by other ways than from extracting it from snakes captured in the wild. A readily available source of this venom will be useful for all pharmacological and biotechnological studies, as well as providing an improved avenue for treatments of snakebites. Here, we show that secretory cells of venom gland can be a good in vitro apparatus to produce venom. We have maintained and morphologically characterized the secretory cells of the Bothrops jararaca venom gland cultured up to 21 days. The isolated cells assemble into acini that growth in size up to 21st day, instead of adhering to the substrate. Bothropasin, a venom metalloprotease, was localized in secretory vesicles by immunoelectron microscopy and venom was also detected in culture medium in a concentration as high as 63 microg/ml. These data show that the acini formed in culture are functionally viable; they can produce and secrete venom. PMID- 16310238 TI - HIV-1 Nef mutations abrogating downregulation of CD4 affect other Nef functions and show reduced pathogenicity in transgenic mice. AB - HIV-1 Nef has the ability to downmodulate CD4 cell surface expression. Several studies have shown that CD4 downregulation is required for efficient virus replication and high infectivity. However, the pathophysiological relevance of this phenomenon in vivo, independently of its role in sustaining high virus loads, remains unclear. We studied the impact of the CD4 downregulation function of Nef on its pathogenesis in vivo, in the absence of viral replication, in the CD4C/HIV transgenic (Tg) mouse model. Two independent Nef mutants (RD35/36AA and D174K), known to abrogate CD4 downregulation, were tested in Tg mice. Flow cytometry analysis showed that downregulation of murine CD4 was severely decreased or abrogated on Tg T cells expressing respectively Nef(RD35/36AA) and Nef(D174K). Similarly, the severe depletion of double-positive CD4+CD8+ and of single-positive CD4+CD8- thymocytes, usually observed with Nef(Wt), was not detected in Nef(RD35/36AA) and Nef(D174K) Tg mice. However, both mutant Tg mice showed a partial depletion of peripheral CD4+ T cells. This was accompanied, as previously reported for Net(Wt) Tg mice, by the presence of an activated/memory like phenotype (CD69+, CD25+, CD44+, CD45RB(Low), CD62(Low)) of CD4+ T cells expressing Nef(RD35/36AA) and to a lesser extent Nef(D174K). In addition, both mutants retained the ability to block CD4+ T cell proliferation in vitro after anti-CD3 stimulation, but not to enhance apoptosis/death of CD4+ T cells. Therefore, it appears that Nef-mediated CD4 downregulation is associated with thymic defects, but segregates independently of the activated/memory-like phenotype, of the partial depletion and of the impaired in vitro proliferation of peripheral CD4+ T cells. Histopathological assessment revealed the total absence of or decrease severity and frequency of organ AIDS-like diseases (lung, heart and kidney pathologies) in respectively Nef(RD35/36AA) and Nef(D174K) Tg mice, relative to those developing in Nef(Wt) Tg mice. Our data suggest that the RD35/36AA and D174K mutations affect other Nef functions, namely those involved in the development of lung and kidney diseases, in addition to their known role in CD4 downregulation. Similarly, in HIV-1-infected individuals, loss of CD4 downregulation by Nef alleles may reflect their lower intrinsic pathogenicity, independently of their effects on virus replication. PMID- 16310239 TI - Modelling of coliform removal in 186 facultative and maturation ponds around the world. AB - The paper presents a very extensive evaluation of the coliform decay in facultative and maturation ponds, based on data from 186 different ponds in the world. The ponds encompass a very wide diversity in terms of physical and operating conditions, covering most situations encountered in practice. The median values for the coliform removal efficiencies were 1.8 log units (98% removal) for primary facultative ponds, 1.0 log units for secondary facultative ponds (90% removal) and 1.2 log units (94% removal) for each maturation pond in the series. Two equations to be used for design purposes were derived for estimating the die-off coefficient K(b) (dispersed flow, 20 degrees C) in facultative and maturation ponds. The first equation led to a slightly better fitting with the observed logarithm of the effluent coliform concentrations (R2 = 0.874), and related K(b) with the pond detention time t and depth H (K(b) = 0.682 H(-1.286) t(-0.103)). The other equation also led to a satisfactory fitting (R2 = 0.845), but was slightly simpler, depending only on the pond depth (K(b) = 0.549 H(-1.456)). PMID- 16310240 TI - A coupled photocatalytic-biological process for degradation of 1-amino-8-naphthol 3, 6-disulfonic acid (H-acid). AB - There has been growing emphasis on the development of coupled treatment systems (e.g., advanced oxidation-biological) for treating poorly biodegradable wastewater. An attempt has been made in the present study to couple photocatalytic (TiO2/UV) pretreatment with conventional activated sludge process to achieve improvement in the biodegradation of H-acid. The combination of titanium dioxide and UV light has been known to generate strong oxidants that degrade several organic pollutants into carbon dioxide via the formation of some intermediates. The intermediates formed may undergo biodegradation readily. Accordingly, photodegradation experiments were carried out initially at an optimized TiO2 dose and the minimum pretreatment time required for transforming H acid was identified. For this purpose, UV-vis spectrophotometry and high performance liquid chromatography (HPLC) were extensively used. Subsequently, it was attempted to biodegrade untreated and pretreated H-acid using activated sludge from the textile industry acclimatized to H-acid. It was found that photocatalytic pretreatment of H-acid for 30 min, during which period approximately 8-10% chemical oxygen demand (COD) removal occurred, can be coupled to second-stage biological treatment for achieving enhanced biodegradation of H acid. PMID- 16310241 TI - Selective removal of arsenate from drinking water using a polymeric ligand exchanger. AB - The new maximum contaminant level (MCL) of 10 microg/L for arsenic in the US drinking water will take effect on January 22, 2006. The compliance cost is estimated to be approximately dollar 600 million per year using current treatment technologies. This research aims to develop an innovative ion exchange process that may help water utilities comply with the new MCL in a more cost-effective manner. A polymeric ligand exchanger (PLE) was prepared by loading Cu2+ to a commercially available chelating ion exchange resin. Results from batch and column experiments indicated that the PLE offered unusually high selectivity for arsenate over other ubiquitous anions such as sulfate, bicarbonate and chloride. The average binary arsenate/sulfate separation factor for the PLE was determined to be 12, which were over two orders of magnitude greater than that (0.1-0.2) for commercial strong-base anion (SBA) exchangers. Because of the enhanced arsenate selectivity, the PLE was able to treat approximately 10 times more bed volumes (BVs) of water than commonly used SBA resins. The PLE can operate optimally in the neutral pH range (6.0-8.0). The exhausted PLE can be regenerated highly efficiently. More than 95% arsenate capacity can be recovered using approximately 22 BVs of 4% (w/w) NaCl at pH 9.1, and the regenerated PLE can be reused without any capacity drop. Upon treatment using FeCl3, the spent brine was recovered and reused for regeneration, which may cut down the regenerant need and reduces the volume of process waste residuals. The PLE can be used as a highly selective and reusable sorbent for removal of arsenate from drinking water. PMID- 16310242 TI - Nowcast modeling of Escherichia coli concentrations at multiple urban beaches of southern Lake Michigan. AB - Predictive modeling for Escherichia coli concentrations at effluent-dominated beaches may be a favorable alternative to current, routinely criticized monitoring standards. The ability to model numerous beaches simultaneously and provide real-time data decreases cost and effort associated with beach monitoring. In 2004, five Lake Michigan beaches and the nearby Little Calumet River outfall were monitored for E. coli 7 days a week; on nine occasions, samples were analyzed for coliphage to indicate a sewage source. Ambient lake, river, and weather conditions were measured or obtained from independent monitoring sources. Positive tests for coliphage analysis indicated sewage was present in the river and on bathing beaches following heavy rainfall. Models were developed separately for days with prevailing onshore and offshore winds due to the strong influence of wind direction in determining the river's impact on the beaches. Using regression modeling, it was determined that during onshore winds, E. coli could be adequately predicted using wave height, lake chlorophyll and turbidity, and river turbidity (R2 = 0.635, N = 94); model performance decreased for offshore winds using wave height, wave period, and precipitation (R2 = 0.320, N = 124). Variation was better explained at individual beaches. Overall, the models only failed to predict E. coli levels above the EPA closure limit (235 CFU/100 ml) on five of eleven occasions, indicating that the model is a more reliable alternative to the monitoring approach employed at most recreational beaches. PMID- 16310243 TI - Use of cholinesterase activity as an indicator for the effects of combinations of organophosphorus pesticides in water from environmental sources. AB - Organophosphorus pesticides (OPs) are commonly detected in agricultural products, animal-derived foodstuffs, and environmental samples. Until now, the focus of research has been to evaluate the adverse effect of a single OP. While each OP may be present at concentrations under recognized as "no observed adverse effect level (NOAEL)", the combined effects of multiple OPs present at these low concentrations have not been sufficiently studied. Therefore, we developed an in vitro testing method to evaluate the toxicity of multiple OPs based on the degree of inhibition of cholinesterase (ChE) activity. This method requires only 10 min to complete and no specialized technology. We examined 15 OPs by this method and categorized them into three groups according to the degree of ChE inhibition. A relationship between the OPs' chemical structures and the degree of ChE inhibition emerged with the moiety -P-O-CN- showing the strongest action. The degree of ChE inhibition increased with multiple OPs, and the degree of inhibition seemed to be additive. These results demonstrate that the combined toxicity of multiple OPs present in food or environmental samples is an easily determined and toxicologically relevant measure of overall toxicity of complex OPs mixtures. It is possible to apply this testing method as a monitoring technique in water quality management in order to control OPs. As a result, this method can play the role for the potential risk reduction to the ecosystem and may contribute to the preservation of the environment. PMID- 16310244 TI - Effect of short-term, low dose aspirin supplementation on the activation of pro inflammatory NF-kappaB in aged rats. AB - The basis of our recently proposed "molecular inflammation theory of aging" is that activated inflammatory transcription factors, including versatile NF-kappaB, occur widespread in the organism during aging. NF-kappaB plays a key role in pro inflammatory gene expression, such as cyclooxygenase (COX). Aspirin is one of the most commonly used non-steroidal anti-inflammatory drugs because of its ability to inhibit COX activity. Thus, in the present study, we investigated the effect of short-term, low dose aspirin intake on the modulation of pro-inflammatory NF kappaB activation in old rats. To conduct the study, 24-month-old Fischer 344 rats were supplemented with low dose aspirin (0.015%) for 10 days. Biochemical analyses showed suppressed reactive species (RS) and COX-2 activity. The data also showed that NF-kappaB activation and its associated gene expressions, such as COX-2, iNOS, VCAM-1 and ICAM-1, were all suppressed by the low dose aspirin supplementation through the inhibition of phosphorylation and degradation of IkappaBalpha via the NIK/IKK pathway. Our molecular exploration further revealed that aspirin's suppressive action of NF-kappaB was mediated by its ability to inhibit the nuclear translocation of cytosolic thioredoxin and redox factor-1. These findings showed for the first time that in aged rat short-term low dose dietary aspirin feeding modulates the molecular signal transduction involved in the inflammatory process. PMID- 16310245 TI - Can MRI rule out bladder and rectal invasion in cervical cancer to help select patients for limited EUA? AB - OBJECTIVES: Although invasion of the bladder or rectum is rare in cervical carcinoma, endoscopic assessment of both organs is part of the standard FIGO clinical staging system, with associated increase in cost and risk of complications. Our objective was to evaluate whether MRI could be used to select patients who did not require invasive staging of the bladder or rectum. METHODS: Two observers, blinded to the results of cystoscopy and endoscopic examination of the rectum, retrospectively reviewed the MR images of 112 patients with cervical carcinoma. A 5-point invasion score was used to determine bladder and rectal invasion (1 = no invasion, 5 = definite invasion). A confidence score of 3 or above was used to identify patients with possible bladder or rectal involvement. The results of cystoscopy and endoscopic examination of the rectum were recorded and correlated with the MR findings. RESULTS: MRI was negative for both bladder and rectal invasion in 94/112 patients. Cystoscopy and endoscopic examination of the rectum were confirmed to be normal in all 94 cases. MRI identified 12 patients with possible rectal invasion, 2 confirmed at endoscopy. MRI identified 14 patients with possible bladder invasion, one confirmed at cystoscopy. Using a low threshold cut-off score of >3 to predict invasion resulted in a 100% negative predictive value (NPV) in detection of bladder and rectal invasion. CONCLUSION: The absence of bladder or rectal invasion can be diagnosed with sufficient confidence using an MRI scoring system to safely obviate the need for invasive cystoscopic or endoscopic staging in the majority of patients with cervical cancer. This could potentially lead to a reduction in staging costs and morbidity. PMID- 16310246 TI - Effect of naringin collagen graft on bone formation. AB - Naringin is a flavonoid available commonly in citrus fruits and is also a HMG-CoA reductase inhibitor. Our laboratory compared the amount of new bone produced by naringin in collagen matrix to that produced by bone grafts and collagen matrix. Twenty bone defects, 5 mm x 10 mm were created in the parietal bone of 14 New Zealand White rabbits. In the experimental group, 5 defects were grafted with naringin solution mixed with collagen matrix, 5 defects were grafted with autogenous endochondral bone. In the control groups, 5 defects were grafted with collagen matrix alone (active control) and 5 were left empty (passive control). Animals were killed on day 14 and the defects were dissected and prepared for histological assessment. Serial sections were cut across each defect. Quantitative analysis of new bone formation was made on 150 sections (50 sections for each group) using image analysis. A total of 284% and 490% more new bone was present in defects grafted with naringin in collagen matrix than those grafted with bone and collagen, respectively. No bone was formed in the passive control group. In conclusion, naringin in collagen matrix have the effect of increasing new bone formation locally and can be used as a bone graft material. PMID- 16310247 TI - Effect of biomaterial surface properties on fibronectin-alpha5beta1 integrin interaction and cellular attachment. AB - The ability of fibronectin (Fn) to mediate cell adhesion through binding to alpha(5)beta(1) integrins is dependent on the conditions of its adsorption to the surface. Using a model system of alkylsilane SAMs with different functional groups (X=OH, COOH, NH(2) and CH(3)) and an erythroleukemia cell line expressing a single integrin (alpha(5)beta(1)), the effect of surface properties on the cellular adhesion with adsorbed Fn layers was investigated. (125)I-labeled Fn, a modified biochemical cross-linking/extraction technique and a spinning disc apparatus were combined to quantify the Fn adsorption, integrin binding and adhesion strength, respectively. This methodology allows for a binding equilibrium analysis that more closely reflects cellular adhesion found in stable tissue constructs in vivo. Differences in detachment strength and integrin binding were explained in terms of changes in the adhesion constant (psi, related to affinity) and binding efficiency of the adsorbed Fn for the alpha(5)beta(1) integrins (CH(3) approximately NH(2)0.05), metals were generally highest at Priest Rapids, relative to downriver dams. Semivolatiles, Aroclors, and organochlorine pesticides were below method reporting limits. Radionuclide differences across locations were minor (Bonferroni P>0.05). Whereas Microtox showed little toxicity, Daphnia IQ tests exhibited measurable toxicity at all locations (EC50 = 22 - 78% porewater). Ecotoxicological benchmarks for metals were exceeded at several locations, most notably at Priest Rapids. Except for K 40, radionuclides were below benchmarks. Overall, chemistry and ecotoxicity results suggested that sediments may pose a risk to benthic biota, likely due to metals (derived largely from upriver mining) or factors associated with a reducing environment (e.g., low oxygen, high ammonia). PMID- 16310250 TI - Complement in BuB/BnJ mice revisited: serum C3 levels and complement opsonic activity are not elevated. AB - With the exception of a few strains such as BuB/BnJ, complement activity in most inbred strains of mice is remarkably similar. The BuB/BnJ strain reportedly has high levels of complement activity and elevated serum levels of C3 and other complement proteins. However, we observed that BuB/BnJ mice have serum C3 levels comparable to those seen in C57BL/6, Balb/c and several other strains of inbred mice. More importantly, using bacteria as a substrate for activation and deposition of complement as a direct biological assay to assess serum complement opsonic activity, we found that BuB/BnJ mice do not have elevated complement activity compared to other inbred mouse strains. In contrast hemolytic assays indicate BuB/BnJ sera to be most active. These results indicate that accepted views regarding mouse serum complement activity need to be re-examined. PMID- 16310251 TI - Episomal vectors to monitor and induce somatic hypermutation in human Burkitt Lymphoma cell lines. AB - Somatic hypermutation (SHM) occurs at a specific B-cell differentiation stage, during the germinal centre reaction, and provides a means to diversify and shape the antibody repertoire of the adaptive immune system. Burkitt-Lymphoma (BL) is a germinal centre derived B-cell malignancy. Presumably deregulation of the somatic hypermutation- and/or class switch recombination process causes a translocation between the myc-locus and one of the Ig-loci, which is characteristic for BL. We show here that we developed an episomal-based vector system to monitor and induce AID-dependent somatic hypermutation in human cell lines. Exemplarily, we utilized this system to investigate the hypermutation status of various BL cell lines. Analysis of the influence of Ig-specific cis-regulatory elements for the mutability of a green fluorescent protein (GFP) reporter transgene revealed, that the presence of Ig-enhancers is required for an efficient targeting of a constitutively transcribed GFP transgene. In contrast, a transcriptional inducible GFP transgene was hypermutated by overexpression of activation-induced cytidine deaminase (AID), even in the absence of Ig-specific sequences. This observation verifies that overexpression of AID in non hypermutating BL cell lines as well as the expression of endogenous AID in the hypermutating BL cell line Raji can overcome the target restriction of AID. PMID- 16310252 TI - Neuropsychological correlates of hair arsenic, manganese, and cadmium levels in school-age children residing near a hazardous waste site. AB - A pilot study was conducted to explore the potential associations between hair metal levels and the neuropsychological function and behavior of school-aged children. Thirty-two children, 11-13 years old, were administered a battery of tests that assessed general intelligence, visual-motor skills, receptive language, verbal memory, nonverbal problem-solving, and behavior problems. Parents and teachers rated the children's attention, executive functions, and behavior problems. The concentrations of manganese (Mn), arsenic (As), and cadmium (Cd) were measured in hair samples provided by 31 of the children. The mean hair metal levels were: Mn, 471.5 parts per billion (ppb); As, 17.8 ppb; Cd, 57.7 ppb. Children's general intelligence scores, particularly verbal IQ scores, were significantly related, inversely, to hair Mn and As levels, as were scores on tests of memory for stories and a word list. In some cases, a significant Mn by-As interaction was found. It appeared that it was the low scores of children for whom both Mn and As levels were above the median values in the sample that were responsible for the main effects observed for each metal. No other significant relationships were found. These results suggest the need to study further the neuropsychological correlates of developmental exposure to Mn and As, particularly as a mixture. PMID- 16310253 TI - Synthesis and structure of cycloruthenated carbonyl complexes and their emission, redox and biological properties. AB - An interesting series of mononuclear organoruthenium complexes of formulation [Ru(CO)(PPh3)2(ap-R)] (where ap-R = -H, -Cl, -Me, -OMe, -OEt) have been synthesized from the reaction of five 2-(arylazo)phenol ligands with ruthenium(II) precursor [RuH(Cl)(CO)(PPh3)3] in benzene under reflux. The 2 (arylazo)phenolate ligands behave as dianionic tridentate ligand and are coordinated to ruthenium through C, N and O by dissociation of the phenolic and phenyl proton at the ortho position of the phenyl ring forming two five-membered chelate rings. These complexes have been characterized by elemental analysis, FT IR, 1H NMR and UV-visible spectroscopy. In dichloromethane solution all the metal complexes exhibit characteristic metal-to-ligand charge transfer (MLCT) absorption and emission bands in the visible region. The structures of [Ru(CO)(PPh3)2(ap-H)] and [Ru(CO)(PPh3)2(ap-Cl)] have been determined by X-ray crystallography. Cyclic voltammetric data of all the complexes show a Ru(III)/Ru(II) oxidation and reduction Ru(II)/Ru(I) within the range 0.74-0.84 V and -0.38 to -0.50 V vs saturated calomel electrode (SCE) respectively. The potentials are observed with respect to the electronic nature of substituents (R) in the 2-(arylazo)phenolate ligands. Further, the free ligands and their ruthenium complexes have also been screened for their antibacterial and antifungal activities, which have shown great promise in inhibiting the growth of both gram +ve and gram -ve bacteria Staphylococcus aureus and Escherichia coli and fungus Candida albicans and Aspergillus niger. These results made it desirable to delineate a comparison between free ligands and their complexes. PMID- 16310254 TI - Prosthetic vascular graft infection: a risk factor analysis using a case-control study. AB - OBJECTIVES: Several factors have been anecdotally reported to increase the risk of prosthetic vascular graft infection (PVGI), a serious complication with high mortality and limb amputation rates. The goal of this study is to confirm purported risk factors using a well-designed statistical model. METHODS: Patients undergoing vascular graft placement at Mayo Clinic Rochester between January 1, 1982 and December 31, 2002 were retrospectively evaluated, excluding redo cases. PVGI cases had to meet one of the following criteria: perigraft air or fluid >8 weeks postoperatively, positive Gram stain or cultures of graft or perigraft material, intraoperative gross purulence, failure of graft incorporation, or exposed graft. Each case had two controls matched for age, gender and date of surgery. Duration of follow-up for a given control had to be at least equal to the time to infection of the corresponding case. Demographic and other clinical data were collected for each patient. RESULTS: Fifty-one cases and 102 controls met the inclusion criteria. Median age was 68 years. Staphylococcal species were identified in 45% of the total number of cases, and 60% of cases with a recovered organism. Univariate analysis identified groin incision, wound complication and wound infection as significant risk factors for PVGI (p<0.005 for all three factors). After adjusting for age and surgery date, multivariate analysis confirmed the statistical significance of groin incision and wound infection as risk factors for PVGI. CONCLUSIONS: This is the first published risk factor analysis that includes statistical evaluation in a case-control study of PVGI. Groin incision, wound complication, and wound infection are significantly associated with developing PVGI. These findings will assist in identifying patients at increased risk of developing subsequent infection. SUMMARY: In order to confirm purported risk factors of prosthetic vascular graft infections (PVGI), a well designed case-control study was conducted at Mayo Clinic, Rochester, MN. Univariate and multivariate analyses of 51 cases and 102 controls identified three statistically significant risk factors for the development of PVGI: groin incision, wound complication, and wound infection. PMID- 16310255 TI - The effect of acute, chronic, and prenatal ethanol exposure on insulin sensitivity. AB - Ethanol has been considered as a lifestyle factor that may influence the risk of type 2 diabetes mellitus. In healthy adults, acute ethanol consumption results in insulin resistance. Acute ethanol consumption causes insulin resistance selectively in skeletal muscle by an indirect mechanism. Possible mediators include triglycerides (TGs), catecholamines, acetaldehyde, alterations in insulin binding, and hepatic insulin sensitizing substance (HISS). Recent studies in rats showed that acute administration of ethanol causes insulin resistance in a dose dependent manner that is secondary to the blockade of insulin-induced HISS release. Chronic ethanol consumption may improve insulin sensitivity, but the results from the randomized controlled trials are mixed. Differences in ethanol dose, consumption period, and abstention period may account for the discrepant results. Epidemiological studies have suggested that the relationship between ethanol and insulin sensitivity is either an inverted U-shape or a positive linear relationship. Future randomized controlled trials should consider the dose of ethanol and the duration of ethanol consumption and abstention in the experimental design. Chronic prenatal and postnatal (nursing) ethanol exposure results in insulin resistance that is secondary to the absence of HISS release/action with the HISS-independent insulin action and insulin-like growth factor-1 (IGF-1)-mediated glucose disposal action remaining unimpaired. The impaired HISS release may be related to a reduction in hepatic glutathione (GSH) levels. The effect of chronic ethanol consumption on HISS has not been evaluated. PMID- 16310256 TI - A modified technique for high-resolution staining of myelin. AB - This report describes a new modification of the Gallyas method for staining myelin in fixed brain tissue and compares results of multiple myelin-visualizing techniques in normal common marmoset (Callithrix jacchus), normal macaque monkey (Macaca mulatta), and a human with multiple sclerosis. The new modification involves immersion in 10% formalin following impregnation in ammoniacal silver nitrate, and the use of a low concentration of 4% paraformaldehyde in the developer. This improved technique is less sensitive to post-mortem tissue handling, temperature, and minor contaminants, allowing a more straightforward implementation in the laboratory setting. It permits simple user-controlled development of the reaction product to maximize contrast in the area of interest, resulting in high contrast staining not only of large axonal bundles, but also thin fascicles throughout tissue sections. Myelin staining in visual cortex of an Old World monkey and a New World monkey reveals similar patterns in the new myelin silver stain, the Gallyas stain, and myelin basic protein immunohistochemistry. The most heavily myelinated areas occupy the edges of blobs, but neither the most lightly stained nor the most darkly stained areas are always in our outside a blob. This indicates a more complex pattern between myelinated axons and blobs than previously suggested. While the new myelin silver stain, darkfield microscopy, the Luxol Fast Blue stain, the Gallyas stain, and myelin basic protein immunohistochemistry all permit visualization of myelin in the CNS, each technique has its own merits and pitfalls; careful evaluation of individual study requirements would best determine which methods are the most useful. PMID- 16310257 TI - Lower cerebrospinal fluid homovanillic acid levels in depressed suicide attempters. AB - BACKGROUND: Studies suggest that the dopaminergic system is involved in the pathogenesis of major depression, Axis II disorders, and suicidal behavior. Depressed suicide attempters constitute a heterogenous group and important differences may exist between depressed suicide attempters with or without Axis II disorders. Therefore, we compared demographic and clinical parameters, and cerebrospinal fluid (CSF) homovanillic acid (HVA) levels in depressed suicide attempters without comorbid Axis II disorders, depressed non-attempters without comorbid Axis II disorders, and normal controls. METHODS: Thirty-one depressed subjects with a history of a suicide attempt, 27 depressed subjects without a history of a suicide attempt, and 50 healthy controls were included in the study. Subjects with comorbid Axis II disorders were excluded. Demographic and clinical parameters, and CSF HVA levels were examined. RESULTS: The two depressed groups did not differ with regard to depression, aggression, hopelessness, and total hostility scale scores. Depressed suicide attempters had higher current suicidal ideation scores compared to depressed non-attempters. Depressed suicide attempters had lower CSF HVA levels compared to depressed non-attempters (t = 4.4, df = 56, p < 0.0001) and to controls (t = -4.09, df = 79, p < 0.0001). There was no difference in CSF HVA levels between depressed non-attempters and controls (t < 1, df = 75, NS). CONCLUSIONS: Dopaminergic abnormalities are associated with suicidality but not with depression. The variability in the rates of comorbid Axis II disorders and in the prevalence of suicide attempters in different patient populations may affect both clinical and biological results of studies of mood disorders. PMID- 16310258 TI - Feeding and contact call stimulation both induce zenk and cfos expression in a higher order telencephalic area necessary for vocal learning in budgerigars. AB - Stimulation with natural contact calls and feeding were used to assess zenk and fos protein expression in budgerigars (Melopsittacus undulatus), a vocal learning parrot species in which feeding and physical contact often occur in conjunction with vocalization. Although only calls induced gene expression in Field L, the primary telencephalic auditory area, both calls and feeding induced gene expression in the frontal lateral nidopallium (NFl), a brain area in receipt of input from Field L which projects to areas afferent to vocal control nuclei and which is necessary for new call learning. NFl thus appears poised to provide both non-auditory as well as auditory feedback to the vocal system. PMID- 16310259 TI - Novel plant-GARP-like transcription factors in Giardia lamblia. AB - GARP homologues constitute a large family of DNA-binding proteins in plants that may be needed for a variety of key cellular functions including regulation of transcription, phosphotransfer signaling, and differentiation. However, no member of this gene family has been reported to date in yeast, animals, or protozoan parasites. We have identified four genes with putative GARP domains in the Giardia lamblia genome (GARP-like protein or GLP). The glp1 mRNA levels increased slightly during encystation. Epitope-tagged GLP1 localized to both nuclei and the proportion of stained Giardia cells increased by 10-fold during encystation. Recombinant GLP1 specifically bound to both the regulated cwp1 and constitutive ran gene promoters in their double-stranded configurations. The C-terminal region of GLP1 containing the GARP-like domain encoded the DNA binding activity. Mutation analysis revealed that an (A/G)ATCN sequence was required for binding of GLP1 to these promoters. We also found that GLP2 recognized similar binding sites. Using mutated plasmids and transfection assays, we demonstrated that the GLP1/2 binding sites are positive cis-acting elements of the cwp1 and ran gene promoters in both trophozoites and encysting cells. GLP1 is the first GARP family gene found in protozoan parasites. Our results suggest that GLP1 may be an important transcriptional activator and that its binding sites are positive promoter elements for certain Giardia genes. PMID- 16310260 TI - Effects of carvedilol on cardiac cytokines expression and remodeling in rat with acute myocardial infarction. AB - OBJECTIVE: A number of observations suggest that cytokines may be important modulators in the ventricular remodeling process. It is unclear whether carvedilol modulates myocardial pro-inflammatory and anti-inflammatory cytokines expression. We hypothesized that carvedilol could improve ventricular remodeling partly through the modulation of cytokines. The goal of this study was to evaluate the effects of carvedilol on cardiac cytokines expression as well as on myocardial and extracellular matrix remodeling in rats with acute myocardial infarction. METHODS: Rats with AMI induced by left anterior descending branch ligation were randomized to carvedilol and control group which were further compared to sham-operated group. We studied the effects of 4-weeks therapy with carvedilol starting 24 h after infarction on 1) hemodynamics, 2) tissue weights, 3) myocardial cytokines (TNF-alpha, IL-1beta, IL-6, IL-10 and TGF-beta1) expression by semi-quantitative RT-PCR and immunoblotting, 4) matrix metalloproteinases activity by gelatin zymography, 5) collagen expression by immunohistochemistry, 6) myocardium fetal gene (alpha and beta myosin heavy chain) expression. RESULTS: Treatment with carvedilol 1) reduced the pro inflammatory cytokines and fibrogenic cytokine TGF-beta1 levels in myocardium and was associated with the amelioration of the elevated left ventricular diastolic pressure. 2) increased anti-inflammatory cytokine, IL-10 protein expression. 3) reduced matrix metalloproteinases-2 and matrix metalloproteinases-9 activity 4) reduced myocardial collagens 5) did not modify fetal gene re-expression. CONCLUSION: Pro-inflammatory, anti-inflammatory and fibrogenic cytokines are all involved in the process of post-infarction myocardial remodeling. One mechanism underlying the beneficial effects of carvedilol on post-infarction myocardial remodeling may be modulation of the balance between pro- and anti-inflammatory cytokines as well as fibrogenic cytokines and extracellular matrix (ECM) remodeling. PMID- 16310261 TI - Enhanced external counterpulsation improves endothelium-dependent vasorelaxation in the carotid arteries of hypercholesterolemic pigs. AB - BACKGROUND: Enhanced external counterpulsation (EECP) has been demonstrated to be an effective method for the treatment of atherosclerotic vascular disease. However, the exact mechanism underlying the beneficial effects of EECP is not completely clear. We hypothesized that EECP leads to improvement in endothelial function, contributing to its clinical benefits. METHODS: Fifteen male domestic pigs were initially divided into 2 dietary groups: one consumed a normal feeding (NF) of pig chow (n=5), and one consumed a high-fat (HF) pig chow (n=10). After 8 weeks on the NF or HF diet, 5 HF pigs received EECP treatment (HF+EECP) 1 h daily for 6 weeks and the remaining 5 HF pigs continued to be fed by high cholesterol diet. At the end of 6-week EECP treatment, the carotid arterial rings from all of the pigs were harvested. Endothelium-dependent and -independent vasorelaxation to acetylcholine (ACh) and sodium nitroprusside (SNP) were measured in a dose dependent manner. RESULTS: The high fat diet resulted in increase in plasma cholesterol and triglyceride levels (p<0.05). Endothelium-dependent vasorelaxation was decreased in the HF group compared to the NF control (p<0.05). However, EECP treatment partially improved impaired endothelium-dependent vasorelaxation in the HF+EECP group compared to the HF control (p<0.05). Endothelium-independent vasorelaxation was not significantly different among the three groups. CONCLUSIONS: Endothelium-dependent vasorelaxation is impaired in the hypercholesterolemic pigs. EECP treatment significantly improves hypercholesterolemia-induced diminished endothelium-dependent vasorelaxation. It suggests that amelioration in endothelial function may at least in part contribute to the beneficial effects of EECP treatment in clinical practice. PMID- 16310262 TI - Lipid peroxidation is not increased in heart failure patients on modern pharmacological therapy. AB - BACKGROUND: Previous studies support a role of oxygen-free radicals in the development of congestive heart failure (CHF). The aim of this study was to investigate whether lipid peroxidation is increased in CHF patients on modern pharmacological therapy and whether there is a positive correlation between plasma levels of markers of lipid peroxidation and severity of heart failure (HF). Plasma malondialdehyde (MDA) and isoprostanes are often used as markers of lipid peroxidation and oxidative stress. We also studied whether long-term treatment with isosorbide-5-mononitrate (IS-5-MN) in combination with standard HF therapy affects P-MDA levels in patients with evidence of left ventricular (LV) dysfunction following acute myocardial infarction (AMI). MATERIALS AND METHODS: Ninety-two patients with clinical or echocardiographic evidence of LV-dysfunction following AMI were randomized to treatment with either IS-5-MN or placebo. In a subgroup of 83 patients with available plasma MDA, echocardiography, right-heart catherization, and plasma natriuretic peptides were evaluated. Control subjects were 80 healthy blood donors. A second study group consisted of 56 patients with CHF, evaluated with respect to LV function, brain natriuretic peptide and markers of oxidative stress (P-MDA and 8-isoprostane). The second control group comprised 50 healthy subjects. RESULTS: Lipid peroxidation measured by P-MDA and 8 isoprostane was not increased in patients with LV dysfunction treated with standard HF therapy. No positive correlation was found to the severity of HF. Long-term IS-5-MN therapy did not influence P-MDA concentrations. CONCLUSIONS: Although results from many experimental and clinical studies suggest that oxidative stress is increased in HF, this may not be true for patients treated with beta blockers and inhibitors of the renin-angiotensin system. PMID- 16310263 TI - Papillary muscle masquerading as a vegetation. AB - We describe a case of an elderly man with an unusual post-operative complication after mitral valve replacement. The final diagnosis was discovered on reoperation. To our knowledge, this phenomenon has not been described in the literature previously. PMID- 16310264 TI - Relation between peripheral vascular endothelial function and coronary flow reserve in patients with chest pain and normal coronary angiogram. AB - To evaluate the relation between peripheral vascular endothelial function and coronary flow reserve (CFR), we assessed flow mediated dilation (FMD) of brachial artery and the intima-media thickness (IMT) of the carotid artery in 32 subjects (mean age 58+/-9 years, M/F=9:23 ) with chest pain and normal coronary angiogram. The subjects were divided into 2 groups according to CFR >or=2.1 or <2.1 measured with transthoracic echocardiography in distal left anterior descending coronary artery. We found % FMD was decreased in the group with CFR <2.1 than those of CFR >or=2.1 and CFR was correlated with peripheral FMD. However, IMT was not different between two groups. These results suggest that microvascular dysfunction is primarily related to endothelial dysfunction rather than advanced atherosclerosis and because it is a generalized process that involves the whole arteries, the measurement of brachial FMD can be a useful diagnostic tool to evaluate microvascular dysfunction in patients with chest pain and normal coronary angiogram. PMID- 16310265 TI - Angiosarcoma of the right atrium: a diagnostic dilemma. AB - Angiosarcomas of the heart are rare and usually fatal. We describe a case where cardiovascular magnetic resonance imaging was crucial in making the diagnosis. Surgical resection was performed and the diagnosis was confirmed by histology. PMID- 16310266 TI - Acute myocarditis mimicking acute myocardial infarction: a clinical nightmare with forensic implications. AB - Authors present the case of the sudden death of a 30-year-old man, 3 h since his hospitalization by the onset of aspecific chest pain. ECG findings revealed the presence of localized ST segment elevation in precordial leads (V1-V4) and DII DII, and aVF mimicking acute antero-inferior myocardial infarction. A diagnosis of acute antero-inferior myocardial infarction was advanced and the patient introduced to thrombolytic therapy. Suddenly, on ECG monitor, conduction abnormalities were early recorded (ventricular extrasystole) followed by ventricular tachycardia degenerating in fatal ventricular fibrillation. An alleged medical malpractice was sued against the cardiologist. A complete immunohistochemical study was performed. Histologically, the heart presented massive interstitial lymphocytic infiltrate and focal myocytes necrosis. The diagnosis of acute lymphocytic myocarditis was established as the cause of death. PMID- 16310267 TI - Dynamic left ventricular obstruction: a potential cause of angina in end stage renal disease. AB - BACKGROUND: Patients with end stage renal disease (ESRD) often have angina which cannot be explained by coronary artery disease (CAD) alone. Symptoms are often attributed to systolic and diastolic dysfunction, arrhythmia or microvascular disease. This study proposes another potential cause for angina in renal failure, namely dynamic left ventricular obstruction (LVO). METHODS: 125 renal transplant candidates underwent dobutamine stress echocardiography and coronary angiography. LVO was defined as a peak LV outflow tract gradient >50 mm Hg. RESULTS: None of the patients had LVO at rest. 15 (12%) developed obstruction with dobutamine. The mechanism was systolic anterior motion of the mitral valve in 13 and mid cavity obstruction in 2. Of 53 patients with angina, 26% had dynamic LVO compared to only 1% in those without angina (p=0.004). The proportion with severe CAD, parameters of systolic and diastolic function and haemoglobin levels were similar in patients with and without angina. Significantly more patients with obstruction had angina (93% vs 28%, p<0.001). LV end-systolic (p=0.03) and LV end-diastolic (LVEDD, p=0.03) diameter were reduced and LV fractional shortening (LVFS) increased (p= or <0.001) in those with LVO. Haemoglobin levels, septal wall thickness, estimated LV filling pressure, the proportion with a positive DSE and severe CAD were similar in the 2 groups. LVFS was independently associated with the development of significant obstruction (OR 1.12, 95% CI [1.002, 1.244] p=0.04). CONCLUSIONS: Dynamic LVO occurs in 26% of ESRD patients with angina. In these patients, the angina could not be explained by severe CAD, impaired systolic or diastolic function. Such patients have smaller LV cavity size and increased LVFS. PMID- 16310268 TI - Analysis of coronary calcifications versus Framingham and PROCAM risk assessment in patients with a first myocardial infarction. AB - We compared the extent of coronary calcifications as quantified by electron beam tomography (EBT) to the predicted 10-year cardiovascular event risk based on Framingham and PROCAM algorithms in patients with a first myocardial infarction (MI). METHODS: In 156 patients (56.7 +/- 22 years), EBT was performed <4 weeks after MI. Coronary calcifications were quantified using the "Agatston Score" and age-related calcium percentiles were determined. The predicted 10-year event risk was determined using Framingham and PROCAM algorithms. RESULTS: Coronary calcifications were present in 148 patients with a mean "Agatston Score" of 589.2 +/- 976. The "Agatston Score" was >400 in 44% and >90th percentile in 42% of the patients. Framingham risk was >20% in 32% and PROCAM risk was >20% in 28%. CONCLUSION: Coronary calcifications were detected in the vast majority of patients with a first MI, independent from the patient's age. The determination of age-related percentiles was superior to the Framingham or PROCAM algorithm. PMID- 16310269 TI - Detection of Listeria monocytogenes and the toxin listeriolysin O in food. AB - Listeria monocytogenes is an emerging bacterial foodborne pathogen responsible for listeriosis, an illness characterized by meningitis, encephalitis, and septicaemia. Less commonly, infection can result in cutaneous lesions and flu like symptoms. In pregnant women, the pathogen can cause bacteraemia, and stillbirth or premature birth of the fetus. The mortality rate for those contracting listeriosis is approximately 20%. Currently, the United States has a zero tolerance policy regarding the presence of L. monocytogenes in food, while Canada allows only 100 cfu/g of food. As such, it is essential to be able to detect the pathogen in low numbers in food samples. One of the best ways to detect and confirm the pathogen is through the detection of one of the virulence factors, listeriolysin O (LLO) produced by the microorganism. The LLO-encoding gene (hlyA) is present only in virulent strains of the species and is required for virulence. LLO is a secreted protein toxin that can be detected easily with the use of blood agar or haemolysis assays and it is well characterized and understood. This paper focuses on some of the common methods used to detect the pathogen and the LLO toxin in food products and comments on some of the potential uses and drawbacks for the food industry. PMID- 16310270 TI - Evaluation of molecular methods to determine enterotoxigenic status and molecular genotype of bovine, ovine, human and food isolates of Staphylococcus aureus. AB - This study evaluated the use of PFGE and single enzyme AFLP techniques for the determination of the genetic relationships between Staphyloccocus aureus isolates from human, bovine, ovine and food related sources and reports the prevalence of 'classic' (sea to see) and 'new' (seg, seh, sei, sej, sem, sen and seo) staphylococcal enterotoxin (se) genes in 92 S. aureus strains. A sub-set of the se genotyping results was confirmed by ELISA and the presence of SE toxin determined in isolates from different sources. A 100% correlation was observed, between detection of enterotoxin genes sea-see and expression of corresponding enterotoxin proteins in vitro. The se genotyping data generated from 90 of the S. aureus isolates showed that many of the S. aureus strains producing identical se genotypes correlated with both AFLP and PFGE pattern types. However, single enzyme AFLP technique did not possess the discriminatory power of the PFGE method, but similar clonal relationships were observed by both techniques in many of the isolates tested. Results reported here include the first comprehensive study using a single enzyme AFLP technique to investigate the genetic background of S. aureus isolates from a wide distribution including animal, human and food related sources. PMID- 16310271 TI - Establishment of an experimental system allowing immobilization of proteins on the surface of Bacillus subtilis cells. AB - Gram-positive bacteria code for one or more enzymes termed sortases which catalyze the covalent anchoring of substrate proteins on their cell wall. They recognize an amino acid sequence designated sorting motif, present close to the C terminal end of the substrate proteins, cleave within this motif and catalyze anchoring of the polypeptide chain to the peptide crossbridge linking the peptidoglycan strands in a transpeptidation reaction. Bacillus subtilis has been reported to code for two different sortases but the sorting sequences recognized by them are yet unknown. To be able to immobilize proteins on the surface of B. subtilis cells, we introduced the srtA gene coding for sortase A of Listeria monocytogenes with the known sorting motif (LPXTG) into B. subtilis. L. monocytogenes and B. subtilis share the same peptide crossbridge. Next, we fused the coding region of an alpha-amylase gene to the C-terminal region of Staphylococcus aureus fibronectin binding protein B containing the sorting motif. Covalent linkage could be proven by treatment of the cells with lysozyme and by immunofluorescence microscopy. Up to 240,000 molecules of alpha-amylase could be immobilized per cell, 24 times more than previously reported for other bacterial species. To study the influence of the distance between the sorting motif and the C-terminus of alpha-amylase on the activity of the enzyme, the length of the spacer was varied. It turned out that the highest activity was measured with a spacer length of 123 amino acid residues. PMID- 16310272 TI - Nitrification performance and microbial community dynamics in a submerged membrane bioreactor with complete sludge retention. AB - A submerged membrane bioreactor (MBR) supplied with inorganic ammonium-bearing wastewater (NH(4)(+)-N, 500 mgl(-1)) was operated for 260 days without sludge purge under decreased hydraulic retention times (HRT) through six steps (from 30 to 5h). Almost complete nitrification was obtained at a volumetric loading rate (VLR)1.2g NH(4)(+)-Nl(-1)day(-1). The sludge nitrification activities were evaluated at each stage. The specific ammonium oxidizing rate (SAOR) decreased from the initial 0.45 to 0.15 kg NH(4)(+)-Nkg(-1)MLSSday(-1) in the last four stages, while the specific nitrate forming rate (SNFR) increased from 0.17 to 0.39 kg NO(3)(-)-Nkg(-1)MLSSday(-1) at the third stage, and then decreased to below 0.1 kg NO(3)(-)-Nkg(-1)MLSSday(-1) from the fourth stage. Microbial population dynamics was investigated by a combination of the MPN method, fluorescence in situ hybridization (FISH) and quinone profiles. During the experiment, although the MLSS increased gradually from 4.5 to 11.5 gl(-1), the number of ammonia-oxidizing bacteria (AOB) decreased from 10(9)l(-1) at the third stage to 10(7)l(-1) in the last two stages, and that of nitrite-oxidizing bacteria (NOB) decreased gradually from 10(8)l(-1) at the second stage (HRT of 20 h) to the final 10(5)l(-1). FISH results showed that the active cells decreased gradually with time from about 60 to 20% in the last two stages, and most of sludge was inert cells. The sum of nitrifiers occupied only about 10% of the total bacteria number in the last stage even though only ammonium-bearing inorganic wastewater was fed in. Nitrosomonas sp. and Nitrospira sp. were confirmed by FISH as the dominant nitrifying genera responsible for ammonia and nitrite oxidation, respectively. In the mean time, a small ratio of Nitrobacter sp. also existed in the system. FISH analysis matched better with the batch activity test results than did the MPN techniques. Quinone profiles revealed that the dominant ubiquinone was ubiquinone-8 (UQ-8), ranging from 84 to 66%, followed by UQ-10 of 7-13%, UQ-7 of 3-5% and UQ-9 of 1.6-2.6%. The dominant menaquinone in the MBR was menaquinone-7 (MK-7) followed by MK-6, MK-8 and MK-8 (H(2)). With the prolongation of operation, the percentage of menaquinones increased from 8 to 14%. The use of the polyphasic approach gave some new insight on variations of microbial community structures. PMID- 16310273 TI - Effect of lpdA gene knockout on the metabolism in Escherichia coli based on enzyme activities, intracellular metabolite concentrations and metabolic flux analysis by 13C-labeling experiments. AB - The lipoamide dehydrogenase (LPD) encoded by lpdA gene is a component of the pyruvate dehydrogenase complex (PDHc), alpha-ketoglutarate dehydrogenase (AKGDH) and the glycine cleavage multi-enzyme (GCV) systems. In the present study, cell growth characteristics, enzyme activities and intracellular metabolite concentrations were compared between the parent strain Escherichia coli BW25113 and its lpdA knockout mutant in batch and continuous cultures. The lpdA knockout mutant produced significantly more pyruvate and L-glutamate under aerobiosis. Some D-lactate and succinate also accumulated in the culture broth. Based on the investigation of enzyme activities and intracellular metabolite concentrations, acetyl-CoA was considered to be formed by the combined reactions through pyruvate oxidase (PoxB), acetyl-CoA synthetase (Acs) and acetate kinase (Ack) phosphoacetyltransferase (Pta) in the lpdA mutant. The effect of the lpdA gene knockout on the intracellular metabolic flux distributions was investigated based on 1H-13C NMR spectra and GC-MS signals obtained from 13C-labeling experiment using the mixture of [U-13C] glucose, [1-13C] glucose, and naturally labeled glucose. Flux analysis of the lpdA mutant indicated that the Entner-Doudoroff (ED) pathway and the glyoxylate shunt were activated. The fluxes through glycolysis and oxidative pentose phosphate (PP) pathway (except for the flux through glucose-6-phosphate dehydrogenase) were slightly downregulated. The TCA cycle was also downregulated in the mutant strain. On the other hand, the fluxes through the anaplerotic reactions of PEP carboxylase, PEP carboxykinase and malic enzyme were upregulated, which were consistent with the results of enzyme activities. Furthermore, the influence of the poxB gene knockout on the growth of E. coli was also studied because of its similar function to PDHc which connects the glycolysis to the TCA cycle. Under aerobiosis, a comparison of lpdA mutant and poxB mutant indicated that PDHc is the main enzyme which catalyzes the reaction from pyruvate to acetyl-CoA in the parent strain, while PoxB plays a very important role in the PDHc-deficient strain. PMID- 16310274 TI - Rapid deswelling of semi-IPNs with nanosized tracts in response to pH and temperature. AB - Porous linear-polymer penetrating networks (semi-IPNs) were prepared by the radical polymerization of acrylic acid (AAc) inside porous poly(N isopropylacrylamide) hydrogels with "nano-sized tract networks (nano-tracts)." The deswelling behavior was investigated by measuring the time course of the swelling ratio change of the semi-IPNs just after the temperature or pH was changed. Although the incorporation of PAAc linear chains into the PNIPAAm hydrogel networks should facilitate deswelling, nonporous semi-IPNs showed only a slow thermoresponse under low pH conditions, and a slow pH-response under a high temperature. On the other hand, the porous semi-IPNs showed a rapid deswelling in response to either a change in pH or temperature, regardless of the environment, presumably due to the nano-tracts through which the water was rapidly released from the hydrogel without any interference by network shrinking. The novel network design of these nano-tracted semi-IPNs may be useful for high performance, dual molecule-release functions. PMID- 16310275 TI - Poly I:C prevents T cell-mediated hepatitis via an NK-dependent mechanism. AB - BACKGROUND/AIMS: T cell immune responses play key roles in the pathogenesis of viral hepatitis, and innate immunity is known to be also activated during this process, however, the effects of innate immunity activation on T cell-mediated hepatitis remain obscure. Here we examined the effect of the activation of NK cells induced by toll-like receptor 3 (TLR3) ligand, polyinosinic-polycytidylic acid (poly I:C), on concanavalin A (Con A)-induced T cell-mediated liver injury. METHODS: Mice received nontoxic intraperitoneal poly I:C injection before Con A intravenous administration. The liver injury was examined by measuring serum transaminase and pathology, and the function of hepatic lymphocytes was detected by FACS analysis. RESULTS: Poly I:C pretreatment protected against T cell mediated hepatitis, as evidenced by decreased mortality, hepatic necrosis, serum transaminase levels and inflammatory cytokines (IL-4, IFN-gamma). The protective effect of poly I:C was diminished in NK-depleted mice, which could be partially restored by adoptive transfer of NK cells. Administration of poly I:C caused NKT and T cell apoptosis via enhancing expression of Fas protein on these cells and expression of Fas ligand on NK cells. CONCLUSIONS: These findings suggest that activation of NK cells by poly I:C prevents Con A-induced T cell-hepatitis via downregulation of T/NKT cells and subsequent reduction of inflammatory cytokines. PMID- 16310276 TI - Up-regulation of the anti-inflammatory adipokine adiponectin in acute liver failure in mice. AB - BACKGROUND/AIMS: Recent reports suggest that the adipose tissue and adipokines are potent modulators of inflammation. However, there is only scarce knowledge on the functional role and regulation of endogenous adiponectin in non-fat tissues such as the liver under conditions of acute inflammation. METHODS: In the present study, we investigated adiponectin expression in healthy murine liver tissue and under inflammatory conditions in vivo. RESULTS: Adiponectin mRNA was readily detectable in healthy liver tissue and further increased in ConA-mediated acute liver failure. Adiponectin protein expression was mainly found in hepatic endothelial cells. In vitro adiponectin mRNA expression was detectable in several cell types, including primary hepatic sinusoidal endothelial cells, stellate cells, and macrophages. Mice pretreated with adiponectin before ConA administration developed reduced hepatic injury as shown by decreased release of transaminases and reduced hepatocellular apoptotis. Of note, TNF-alpha levels were not affected by adiponectin, whereas IL-10 production was increased. Neutralisation of IL-10 diminished the protective effect of adiponectin. CONCLUSIONS: Adiponectin expression is up-regulated in ConA-mediated acute liver failure. Therefore, adiponectin might play a role in the control and limitation of inflammation in the liver. Moreover, our data suggest a role for IL-10 in adiponectin-mediated hepatoprotection. PMID- 16310277 TI - Hepatic environment elicits monocyte differentiation into a dendritic cell subset directing Th2 response. AB - BACKGROUND/AIMS: Dendritic cells (DCs), which play a critical role during immune response, could present alternative differentiation patterns depending on tissue microenvironment. Our aim was to examine the influence of hepatic microenvironment on human monocyte differentiation into DCs. METHODS: Cytology, immunophenotyping, cytokine production and T-cell activation were analyzed in DCs differentiated from human monocytes co-cultured with rat liver epithelial cells (RLEC) or human cells from various tissue origins and compared to control DCs obtained on plastic with GM-CSF/IL-4. RESULTS: RLEC environment promotes DC differentiation in the presence of IL-4 without GM-CSF. These DCs evidence similar expression of MHC-II, co-stimulatory and adhesion molecules than control DCs, but distinct lineage markers defining a CD11c+/CD14+/CD123+ DC subset. This phenotype is common to DCs from RLEC and human liver environment and differs from that obtained with skin or intestine environments. Functionally, they produce IL 10 but not IL-12p70 and favor IL-4/IL-10 secretion by T-cells rather than IFN gamma. CONCLUSIONS: Our results confirm that tissue niches modulate DC differentiation and demonstrate that hepatic environment influences monocyte differentiation into a DC subset directing Th2 response, a key data for understanding the specialized immune response in liver. They also make RLEC co culture system useful for studying liver DC functions. PMID- 16310278 TI - Suppression of hepatocellular carcinoma growth in mice via leptin, is associated with inhibition of tumor cell growth and natural killer cell activation. AB - BACKGROUND/AIMS: Leptin exerts potent immune modulatory properties. The aim of this study was to determine leptin's anti-tumor effect in a murine model of hepatocellular carcinoma (HCC). METHODS: In vivo, athymic nude mice were transplanted with Hep3B cells, followed by daily leptin administration for 6 weeks. RESULTS: Leptin administration induced a significant reduction in tumor size, improved survival rate, and was associated with a significant increase in peripheral natural killer (NK) cell number. Splenocytes from leptin-treated mice featured decreased expression of CIS mRNA. SCID mice featured a similar leptin associated tumor suppression. In contrast, NK-deficient SCID-beige mice developed larger tumors which were unresponsive to leptin. NK cells incubated in vitro with increasing doses of leptin demonstrated increased cytotoxicity and proliferation. Incubation of leptin with hepatoma cells induced a dose-dependent reduction in proliferation, suggesting a direct anti-tumor effect. Leptin induced increased mRNA expression of STAT2 and SOCS1 on HCC cell lines. CONCLUSIONS: Leptin administration induces a significant suppression of human HCC. This effect is mediated by induction of natural killer cell proliferation and activation, along with direct inhibition of tumor growth. Decreased NK expression of inhibitory CIS and over-expression of the antiproliferative STAT2 and SOCS1 proteins in HCC lines may underline the anti-cancerous effects of leptin. PMID- 16310279 TI - Immunostimulatory CpG-oligodeoxynucleotides (CpG-ODN) induce early hepatic injury, but provide a late window for protection against endotoxin-mediated liver damage. AB - BACKGROUND/AIMS: An impaired immunologic response to infection has been recognized as a major defect in the pathogenesis of sepsis and multi-organ failure. Sepsis-associated liver dysfunction and damage are main determinants for the course of the disease. CpG-motif-containing DNA-sequences (CpG-ODN) were previously shown to confer protection in models of infection by stimulating both innate and specific immune responses. Herein, we studied the effect of CpG-ODN in lipopolysaccharide (LPS)-associated hepatotoxicity. METHODS: Sprague Dawley rats pre-treated at day 6 with either CpG-ODN or inert DNA were challenged with E. coli LPS and subsequently studied for liver injury at 6 and 16 h using in vivo fluorescence microscopy and immunohistochemistry. Western blot protein analysis served for assessment of expression of TLR4, TNF receptor-associated factor 6 (TRAF6), NFkappaB and caspase-3. To evaluate CpG-ODN effects during non-septic conditions, additional animals were solely exposed to CpG-ODN and studied after 1 and 6 days. RESULTS: CpG-ODN application induced marked hepatic microcirculatory deterioration and liver dysfunction at day 1, however, with almost complete recovery to normal at day 6. Interestingly, CpG-ODN pre-treatment decreased LPS induced leukocyte-endothelial cell interaction, sinusoidal perfusion failure and caspase-3-dependent apoptotic cell death. Although Kupffer cell phagocytic activity was not affected, CpG-ODN pre-treatment in LPS-challenged animals attenuated hepatic protein expression of TRAF6 and NFkappaB and increased TLR4 by almost 100%. CONCLUSIONS: CpG-containing DNA-sequences induce early hepatic injury, but mediate long-term protection against LPS hepatotoxicity. The mechanism of protection is based on the induction of cross-tolerance, probably via inhibition of the downstream TRAF6-NFkappaB signaling pathway and upregulation of the TLR4 surface receptor. PMID- 16310280 TI - The finest branches of the biliary tree might induce biliary vascularization necessary for biliary regeneration. AB - BACKGROUND/AIMS: The finer branches of the biliary tree play an important role in biliary regeneration. They are consistently escorted by microvessels. Defects in the vascularization of these structures could impair bile duct regeneration. Therefore, we investigated the pattern of the escorting microvessels during the development of bile duct loss in the human liver, using chronic rejection as a model. METHODS: The number of interlobular bile ducts, bile ductules and extraportal biliary cells with and without escorting microvessels and the expression of VEGF-A were studied in follow-up biopsies of 12 patients with chronic rejection and 16 control patients with acute rejection without progression to chronic rejection. RESULTS: The controls showed a proliferation of bile ductules at 1-week and 1-month. Proliferation of bile ductules without microvessels preceded proliferation of bile ductules with microvessels. Proliferation of the microvascular compartment followed biliary proliferation. This sequence of events was not observed in the chronic rejection group, in which all biliary structures decreased in time. VEGF-A expression was increased at 1 week and 1-month in both groups. CONCLUSIONS: An immediate proliferative response of the finer branches of the biliary tree followed by proliferation of the microvascular compartment after biliary injury seems to be a prerequisite for bile duct regeneration. PMID- 16310281 TI - Expected increase in hepatitis C-related mortality in Egypt due to pre-2000 infections. AB - BACKGROUND/AIMS: Egypt has the highest prevalence of Hepatitis C Virus (HCV) in the world, apparently due to mass parenteral antischistosomal therapy. Estimating the future burden of HCV in Egypt is important to support health policies to combat the epidemic. METHODS: A previous back calculation model was adapted to the situation in Egypt. It combines a model of the natural history of HCV infections with available epidemiological data to back calculate the past HCV incidence from observed 1980-1999 hepatocellular carcinoma (HCC) mortality. In turn, the HCV-related mortality burden is projected in the future due to pre-2000 infections. RESULTS: Compared with the observed number of HCC deaths in 1999, the model predicts a 3.5-fold increase in this mortality in the next 20 years. Globally, the model predicts a 2.4-fold increase in the HCV-related mortality. These predictions do not take into account the new infections that may occur after 2000, which would still increase the estimated future mortality burden. CONCLUSIONS: HCV-related mortality is expected at least to double in the next 20 years. The use of antiviral therapies can lower these predictions. Efficient prevention policies are needed to avoid these predictions being exceeded. PMID- 16310282 TI - Detection of clustered microcalcifications in small field digital mammography. AB - The most frequent symptoms of ductal carcinoma recognised by mammography are clusters of microcalcifications. Their detection from mammograms is difficult, especially for glandular breasts. We present a new computer-aided detection system for small field digital mammography in planning of breast biopsy. The system processes the mammograms in several steps. First, we filter the original picture with a filter that is sensitive to microcalcification contrast shape. Then, we enhance the mammogram contrast by using wavelet-based sharpening algorithm. Afterwards, we present to radiologist, for visual analysis, such a contrast-enhanced mammogram with suggested positions of microcalcification clusters. We have evaluated the usefulness of the system with the help of four experienced radiologists, who found that it significantly improves the detection of microcalcifications in small field digital mammography. PMID- 16310283 TI - Dendrimer-drug interactions. AB - The interaction between drugs and dendrimers is reviewed with particular reference to the entrapment of drugs within the dendrimer architecture and the electrostatic and covalent complexation of drugs to the dendrimer surface. The application of dendrimer-drug complexation in the enhancement of drug solubility and bioavailability and the use of the complexes as vehicles for the controlled release of drugs and drug targeting is discussed. PMID- 16310284 TI - Dendrimers in gene delivery. AB - Dendrimers have unique molecular architectures and properties that make them attractive materials for the development of nanomedicines. Key properties such as defined architecture and a high ratio of multivalent surface moieties to molecular volume also make these nanoscaled materials highly interesting for the development of synthetic (non-viral) vectors for therapeutic nucleic acids. Rational development of such vectors requires the link to be made between dendrimer structure and the morphology and physicochemistry of the respective nucleic acid complexes and, furthermore, to the biological performance of these systems at the cellular and systemic level. The review focuses on the current understanding of the role of dendrimers in those aspects of synthetic vector development. Dendrimer-based transfection agents have become routine tools for many molecular and cell biologists but therapeutic delivery of nucleic acids remains a challenge. PMID- 16310285 TI - Role of melanocortin signaling in the regulation of the hypothalamic-pituitary thyroid (HPT) axis. AB - The melanocortin signaling system is orchestrated by two, independent groups of neurons in the hypothalamic arcuate nucleus with opposing functions that synthesize either alpha-melanocyte stimulating hormone (alpha-MSH) or agouti related protein (AGRP). These neurons exert regulatory control over hypophysiotropic TRH neurons in the hypothalamic paraventricular nucleus (PVN) at least in part through direct, overlapping, monosynaptic projections to the PVN. Alpha-MSH has an activating effect on hypophysiotropic TRH neurons via the phosphorylation of CREB, and when administered exogenously, can completely reverse fasting-induced suppression of TRH mRNA in the PVN. AGRP has a potent inhibitory effect on the hypothalamic-pituitary-thyroid axis in normally fed animals, mediated through actions at melanocortin 4 receptors. Inhibition of the HPT axis by fasting may be explained by inhibition of melanocortin signaling as a result of a reduction in alpha-MSH and increase in AGRP. Neuropeptide Y may also modulate the effects of the melanocortin signaling system during fasting by potentiating the inhibitory actions of AGRP on hypophysiotropic TRH neurons to prevent the phosphorylation of CREB and through direct inhibitory effects on alpha-MSH-producing neurons in the arcuate nucleus. PMID- 16310286 TI - Bioinformatic analysis of neuropeptide and receptor expression profiles during midgut metamorphosis in Drosophila melanogaster. AB - Neuropeptides are important messenger molecules in invertebrates, serving as neuromodulators in the nervous system and as regulatory hormones released into the circulation. Understanding the function of neuropeptides will require the integration of genetic, biochemical, physiological and behavioral information. The advent of DNA microarrays and bioinformatic databases provides a wealth of data describing the expression profiles of thousands of genes during biological processes. One such array catalogs the developmental patterns of gene expression during the metamorphic transformation of the Drosophila midgut. We have mined the data from this experiment to explore changes of expression in genes coding for known neuropeptides, peptide hormones, and their receptors during the metamorphosis of the midgut. We found small but significant changes in the expression of the peptides diuretic hormone, FGLa-type allatostatins, myoinhibiting peptide, ecdysis-triggering hormone, drosokinin and the burs subunit of bursicon, as well as the receptors DAR-2, NPFR1, ALCR-2, Lkr and DH-R. Just as advances have been made in understanding the molecular basis of invertebrate neuropeptide action by analysis of genome projects, data mining of gene expression databases can help to integrate molecular, biochemical and physiological knowledge of biological processes. PMID- 16310287 TI - Modulation of a neuronal network by electrical high frequency stimulation in striatal slices of the rat in vitro. AB - The effects of the GABA(A) receptor antagonist bicuculline, the D2-like receptor antagonist sulpiride and the D1-like receptor antagonist SCH-23390 on the electrical high frequency stimulation (HFS)-evoked gamma-aminobutyric acid (GABA) and dopamine (DA) release were measured from slices of the rat striatum by means of HPLC method with electrochemical detection. HFS with 130Hz stimulated veratridine-activated GABAergic neurons resulting in an increased GABA outflow while DA outflow decreased. In the presence of the GABA(A) receptor antagonist bicuculline extracellular GABA and DA outflow were enhanced. When the competitive dopamine D2-like receptor antagonist S-(-)-sulpiride was added to incubation medium, the HFS-evoked stimulatory effect on GABA outflow declined to values found after veratridine (1microM) without HFS. After co-incubation of sulpiride and the competitive D1-like receptor antagonist R-(+)-SCH-23390, the effect of sulpiride on HFS plus veratridine-evoked GABA outflow was completely reversed. Neither sulpiride nor SCH-23390 had any influence on the effect of HFS on veratridine-induced DA outflow. No effect of HFS on glutamate outflow was observed in all experiments. These results led us to suggest that in our model HFS primarily affects GABAergic neurons. These neurons are embedded in a neuronal network with a GABA-dopamine circuit, and thus, HFS interacts with a neuronal network, not only with one neurotransmitter system or one neuron population. PMID- 16310288 TI - Sparing of age-related macular degeneration in rheumatoid arthritis. PMID- 16310289 TI - [A fatal swallowed fish bone]. PMID- 16310290 TI - [Pancreatic involvement in Wegener's granulomatosis: a rare and difficult diagnosis]. AB - INTRODUCTION: Pancreatic involvement in the course of wegener's granulomatosis is rarely reported. EXEGESIS: We report a case presenting as a pancreatic tumorlike lesion. We also discuss six further cases from the medical literature. Two of them had a tumorlike lesion, three had acute pancreatitis and the last had exocrine pancreatic insufficiency. CONCLUSION: Treatment, association of corticoid and cyclophosphamide, is efficient if it is started quickly. So we think that this unusual manifestation of Wegener's granulomatosis should be recognized. PMID- 16310291 TI - [Demential revealing a Sneddon's syndrom]. PMID- 16310292 TI - [A rare cause of sciatica]. PMID- 16310293 TI - Kinetics of antiviral CD8 T cell responses during primary and post-vaccination secondary bovine respiratory syncytial virus infection. AB - We have measured antiviral CD8 T cells responses in bovine respiratory syncytial virus (bRSV) infected calves that had been immunized with either formalin inactivated (FI) or live-attenuated (L) bRSV, with evidence of immunopathology following challenge of calves vaccinated with FI-bRSV. In all cases, bRSV infection induced potent pulmonary CD8 T cell responses. The kinetics of the post challenge response in L-bRSV immunized animals was accelerated compared to the FI bRSV and PBS groups, suggesting that only the L-bRSV vaccine, and not the FI-bRSV vaccine, had primed memory T cells. The differences between primary and post vaccination secondary infection were very minor, in terms of the proliferation status of pulmonary CD8 T cells. Functional IFN-gamma+ CD8 responses were slightly higher in the FI-bRSV vaccinated animals. Furthermore, the existence of strong IFN-gamma+ CD8 responses in FI-bRSV vaccinated animals after challenge suggests (i) that these IFN-gamma+ responses in FI-bRSV immunized animals do not protect against immunopathology, and (ii) that Th-2 biased responses during bRSV challenge after vaccination with FI-bRSV have a limited impact on the CD8 responses in the bronchoalveolar lavage fluid. Thus, several response patterns (Th-l/Th-2) seem to co-exist during bRSV infection. PMID- 16310294 TI - IL-12-assisted immunization against Listeria monocytogenes using replication restricted VSV-based vectors. AB - Co-administration of IL-12 with vaccine immunogens has proven to be an effective strategy for eliciting potent Th1-biased immunity. Unfortunately, the use of IL 12 as a vaccine component has been limited because it is unstable at ambient temperatures, expensive to produce, and toxic when administered at excessive dosages. Using reverse genetics, we created a recombinant replication-restricted vesicular stomatitis virus that expresses large quantities of an IL-12 fusion protein (VSVDeltaG-IL12F), but can only establish a single round of infection because the genome does not encode the viral glycoprotein (G protein) that is required for viral entry into host cells. Here, we report that immunization of mice with a poorly immunogenic listerial antigen preparation (LMAg) in combination with VSVDeltaG-IL12F elicits potent T cell- and B cell-mediated responses that confer protective listerial immunity. PMID- 16310295 TI - Adverse events after hepatitis A B combination vaccine. AB - In May 2001, the U.S. Food and Drug Administration (FDA) approved Hepatitis A Inactivated and Hepatitis B Recombinant Vaccine (HEPAB) for immunization of adults. From May 2001 to September 2003, the Vaccine Adverse Event Reporting System (VAERS) received 305 reports of adverse events after HEPAB. Many events were similar to those reported after the monovalent hepatitis A and B vaccines. Non-serious events included constitutional symptoms and local reactions. Serious events included neurologic, hepatobiliary, and dermatologic conditions, and detailed medical and epidemiological review did not suggest a clear pattern of evidence supporting a causal relationship with the vaccine, except for injection site reactions and some allergic reactions. PMID- 16310296 TI - INTERESTING CASE: unusual presentation of adenoid cystic carcinoma of the maxillary antrum. PMID- 16310297 TI - INTERESTING CASE: when you can't see the wood: a foreign body pitfall. PMID- 16310298 TI - Glenohumeral joint loading in tetraplegia during weight relief lifting: a simulation study. AB - BACKGROUND: The incidence of shoulder complaints in wheelchair users is high and the etiology is poorly understood. The goal of this study was to examine the effect of lesion level and isolated triceps muscle paresis on the internal load on the shoulder by simulation. METHODS: Kinematic and kinetic profiles from four able-bodied subjects and four subjects with tetraplegia were used as input for an inverse dynamics biomechanical model. The model was modified to simulate lesion level and triceps muscle paresis. FINDINGS: The simulations resulted in a significantly higher (+56%) glenohumeral contact force (P=0.037) for tetraplegic profiles than for able-bodied profiles. The model modifications to simulate lesion level only had a minor effect (+7%) on the calculated glenohumeral contact force. More simulations were successful at lower triceps force levels for tetraplegic profiles compared to able-bodied profiles (P=0.012). The muscle forces at the simulated T1 lesion were not significantly higher in tetraplegic profiles compared to able-bodied profiles. INTERPRETATION: The glenohumeral contact force for the tetraplegic profiles is mainly higher due to different task performance. Model modifications only have a minor effect on the calculated glenohumeral contact force. For able-bodied profiles the triceps force seems to be an important factor. The high internal load at the shoulder recommends new techniques of weight relief lifting and proper training of the arm-shoulder muscles in rehabilitation. PMID- 16310299 TI - Spine loading as a function of lift frequency, exposure duration, and work experience. AB - BACKGROUND: Physiological and psychophysical studies of the effects of lifting frequency have focused on whole-body measurements of fatigue or subjective acceptance of the task and have not considered how spine loads may change as a function of lift frequency or lift time exposure. Our understanding of biomechanical spine loading has been extrapolated from short lifting bouts to the entire work day and may have led us to incorrect assumptions. The objective of this project was to document how spine loading changes as a function of experience, lift frequency, and lift duration while repetitively lifting over the course of an 8-h workday. METHODS: Twelve novice and twelve experienced manual materials handlers performed repetitive, asymmetric lifts at different load and lift frequency levels throughout an 8-h exposure period. Compression, anterior posterior shear, and lateral shear were evaluated over the lifting period using an EMG-assisted biomechanical model. RESULTS: Spinal loads increased after the first 2 h of lifting exposure regardless of the lift frequency. Loading was also greater for the inexperienced subjects compared to experienced lifters. The greatest spine loads occurred at those lift frequencies and weights to which the workers were unaccustomed. INTERPRETATION: Increases in spine loading were tracked back to the changes in muscle recruitment patterns that typically involved increased muscle coactivation. The results emphasize the importance of previous motor programming in defining spine loads during repetitive lifting. These results indicate a very different influence of frequency and lift time exposure compared to physiologic and psychophysical assessments. This study has shown that it is not sufficient to extrapolate from short lift periods to extended exposure periods if the biomechanical loading implications of the task are of interest. PMID- 16310300 TI - Effects of NO2 and NH3 from road traffic on epiphytic lichens. AB - The results of a survey aimed at investigating whether NO2 and NH3 emitted by road traffic can influence lichen diversity, lichen vitality and the accumulation of nitrogen in lichen thalli are reported. For this purpose, distance from a highway in a rural environment of central Italy was regarded as the main parameter to check this hypothesis. The results of the present survey indicated that road traffic is not a relevant source of NH3. On the other hand, NO2 concentrations, although rather low, were negatively correlated with distance from the highway according to a typical logarithmic function. No association between NO2 concentrations and the diversity of epiphytic lichens was found, probably because of the low NO2 values measured. Also bark properties were not influenced by distance from the highway. Accumulation of nitrogen, reduction in the content of chlorophyll a, chlorophyll b and total carotenoids were found in transplanted thalli of Evernia prunastri, but NO2 was not responsible for these changes, which were probably caused by applications of N-based fertilizers. PMID- 16310301 TI - The search for social validation and the sexual behavior of people living with HIV in Rio de Janeiro, Brazil: understanding the role of treatment optimism in context. AB - The primary aim of this qualitative study was to explore the influence of HIV treatment optimism on the sexual behavior of people living with HIV/AIDS (PLWHA) receiving highly active anti-retroviral therapy (HAART) at public health clinics in Rio de Janeiro, Brazil. We also explored the psycho-social dynamics of participants' sexual relationships in order to understand more broadly how these factors influence the sexual behavior of PLWHA and how they shape HAART-related beliefs. Twenty-three semi-structured, in-depth interviews were conducted with three groups: heterosexual women, heterosexual men and men who have sex with men living with HIV who reported being sexually active in the last year and were currently receiving HAART. We found that the availability of HAART was conceptualized as a rationale for unsafe sex among a minority of study participants and that this was more common among men than among women. Specific examples of treatment optimism appeared to be employed as a means to avoid acknowledging how deeper psychosocial issues may have been influencing participant's sexual behavior. Most participants' sexual behavior appeared largely to be a product of their desire for social validation and linked to feelings of shame and denial, including but not limited to HIV. Participants from all three groups expressed a considerable amount of fear and/or anxiety regarding behaviors such as disclosure and condom use as a result of the unexplored conflict between implementing these behaviors and continuing with their strategies for social validation within the context of their sexual relationships. We conclude that short-term information, education and communication interventions surrounding treatment optimism, disclosure and condom use are appropriate and necessary; but that they are not sufficient to address the core challenges to unsafe sex among PLHWA. These deep-rooted psychosocial issues may be better addressed by longer-term individual and group-level opportunities for exploration and critical reflection regarding sense of self and its relationship to social solidarity among PLWHA. PMID- 16310302 TI - The effect of a novel silane blend system on resin bond strength to silica-coated Ti substrate. AB - OBJECTIVES: The adhesive performance of 3-acryloyloxypropyltrimethoxysilane and a silane blend consisting of 3-acryloyloxypropyltrimethoxysilane and 1,2 bis(triethoxysilyl)ethane, adhering an experimental dendrimer-based resin to a silica-coated titanium surface, conditioned by CoJet method, was evaluated in vitro. METHODS: 3-Acryloyloxypropyltrimethoxysilane (1.0 vol%) and a blend of 3 acryloyloxy-propyltrimethoxysilane and 1,2-bis(triethoxysilyl)ethane (both 1.0 vol%) were prepared in acidified 95 vol% ethanol. A commercial 3 methcryloyloxypropyltrimethoxysilane (ESPE Si) was used as control. The silanes were applied onto tribochemically silica-coated titanium surfaces. Fresh silane solution hydrolysis (activation) was monitored by Fourier transform infrared (FTIR) spectrometry. Experimental dendrimer resin stubs were applied and photo polymerized on titanium. The specimens were thermocycled (6000 cycles, 5-55 degrees C). Before measuring shear bond strength of the resin to titanium, surface imaging was carried out with a scanning electron microscope (SEM) with elemental analysis applying energy dispersive X-ray analysis (EDXA). RESULTS: Statistical analysis (ANOVA) showed that the highest shear bond was obtained with 1.0 vol% 3-acryloyloxypropyltrimethoxysilane+1.0 vol% 1,2 bis(triethoxysilyl)ethane blend (5.2+/-1.3 MPa), and the lowest result with fresh commercial 3-methacryloyloxypropyltrimethoxysilane, ESPE Sil (2.1+/-0.8 MPa). 3 Acryloyloxypropyltrimethoxysilane yielded 4.1+/-1.0 MPa. The type of silane affected significantly the shear bond strength (ANOVA, p<0.001). All silanes were activated according to the infrared spectra. The SEM/EDXA analysis revealed an uneven distribution of titanium, silicon, and aluminum on silica-coated Ti surfaces. CONCLUSION: Silanization with 3-acryloyloxypropyltrimethoxysilane might offer better bonding of a novel dendrimer resin to silica-coated titanium than using a pre-hydrolyzed 3-methacryloyloxypropyltrimethoxysilane. PMID- 16310303 TI - Dentist-time expenditure for two different adhesive all-ceramic systems. AB - OBJECTIVES: The purpose of this clinical investigation was to compare a chair side adhesive all-ceramic system to a laboratory processed adhesive all-ceramic system with respect to quality and time expenditure for the dentist. METHODS: The same dentist treated 10 patients, who were each to receive two large posterior single tooth restorations of similar location and extent. One restoration was made in the laboratory by using the IPS Empress system [LAB], the other one was done chair-side by utilizing the Cerec system [CHAIR]. The time expenditure was measured for [LAB] and [CHAIR] and compared by the Wilcoxon signed rank test. The restorations were also evaluated according to the USPHS criteria. RESULTS: The mean time expenditure for the dentist with low-level assistance was 111:03 min [S.D.+/-24:09 min] for [LAB] and 115:31 min [S.D.+/-15:54 min] for [CHAIR]. Time expenditure with medium level assistance for the operator was 100:53 min [S.D.+/ 23:59] for [LAB] and 105:50 [S.D.+/-15:28] for [CHAIR]. Assuming a high level of assistance, the mean time values were 53:11 min [S.D.+/-14:29] for [LAB] and 54:29 min [S.D.+/-09:21] for [CHAIR]. The baseline investigation according to the modified USPHS criteria did not reveal any differences between [CHAIR] and [LAB]. CONCLUSION: There were no statistical significant differences with respect to time expenditure or quality between [LAB] and [CHAIR] in this study. PMID- 16310304 TI - Pregnancy outcome of patients with known celiac disease. AB - OBJECTIVE: Celiac disease is a permanent intolerance to gluten, probably induced by an autoimmune mechanism. Controversy exists regarding the association between celiac disease and infertility, abortions, intra-uterine growth restriction (IUGR) and stillbirths. The present study was designed to investigate pregnancy outcome of patients with celiac disease. METHODS: A retrospective comparison between all pregnancies of women with and without known celiac disease, delivered during the years 1988-2002, was conducted. RESULTS: During the study period there were 48 deliveries of patients with celiac disease and 143,663 pregnancies of patients without known celiac disease. No statistically significant differences were noted between the groups regarding maternal or perinatal outcomes, including fertility treatments (0% among patients with known celiac versus 2.5% among patients without known celiac sprue; p=0.267), recurrent abortions (0 versus 5.2%; p=0.103), perinatal mortality (2.1 versus 1.4%; p=0.668). However, higher rates of labor induction (29.2 versus 11.9%; p<0.001) and IUGR (6.3 versus 2.1%; p=0.042) were found among patients with celiac disease as compared to patients without known celiac disease. CONCLUSION: The course of pregnancy of patients with celiac disease including perinatal outcomes is favorable. Since we found higher rates of IUGR, careful surveillance should be performed for early detection of IUGR. Further, prospective studies should focus on screening for celiac disease among patients presenting with IUGR of an unknown etiology. PMID- 16310305 TI - Nifedipine versus ritodrine for suppression of preterm labor. Comparison of their efficacy and secondary effects. AB - OBJECTIVES: To compare the efficacy of nifedipine and ritodrine in prolonging pregnancy beyond 48 h, 1 week and 36.0 weeks and to evaluate maternal side effects and adverse perinatal outcome. STUDY DESIGN: Non-blinded, randomized controlled trial. Eighty patients with singleton pregnancies admitted for preterm labor with intact membranes between 22 and 35 weeks of gestation were included in the study. Preterm labor was defined as the persistence of at least two symptomatic uterine contractions within a 10 min period during 60 min after admission and despite bed rest. RESULTS: Forty women received oral nifedipine and forty intravenous ritodrine. Two patients, one from each group, were excluded because of loss to follow-up after discharge. Therefore, 39 women in the nifedipine and the ritodrine groups, respectively, were evaluable for the final analysis. Baseline characteristics were comparable in both groups. The percentage of initial response, the speed of onset of action and the rate of successful treatment within 48 h were significantly better in the ritodrine group. However, prolongation of pregnancy beyond 7 days and 36 weeks of pregnancy was similar with a significantly lower rate of side effects in the nifedipine group. CONCLUSIONS: In this small trial, ritodrine provided more effective tocolysis within the first 48 h than nifedipine at the doses used in this study, although with a significantly higher rate of side effects. PMID- 16310306 TI - TEL/ARG induces cytoskeletal abnormalities in 293T cells. AB - We previously identified TEL/ARG as a novel fusion transcript consisting of the oligomerization domain of TEL and the kinase domain of ARG, in a case of acute myeloid leukemia. We report here the existence of an alternatively spliced TEL/ARG transcript lacking part of a F-actin binding domain of ARG, and the phenotype of TEL/ARG expressing 293T cells. In 293T cells, both TEL/ARG forms co localized with the cellular beta-actin and were associated with a morphologic change of the cells, consisting in cell rounding and detachment from the tissue culture plastic. We identified the Rho inhibitor p190RhoGAP, a critical regulator of cellular adhesion, as a target of the aberrant kinase. PMID- 16310307 TI - Equilibrium studies for trimethoprim adsorption on montmorillonite KSF. AB - In this study, the adsorption of trimethoprim (TMP) on montmorillonite KSF was studied under different conditions (pH, ionic strength, temperature). The results indicate that a pH value of 5.04 is optimum value for the adsorption of TMP on KSF. The adsorption kinetics was interpreted using pseudo-first-order kinetic model, pseudo-second-order kinetic model and intraparticle diffusion model. The pseudo-second-order model provides the best correlation with the experimental data of KSF adsorption. The adsorption data could be fitted with Freundlich, Langmuir and Dubinin-Radushkevich equation to find the characteristic parameters of each model. It was found that linear form of Langmuir isotherm seems to produce a better model than linear form of Freundlich equation. From the Langmuir and Freundlich equation, the adsorption capacity values raised as the solution temperature decreased. From DR isotherm, it was also determined that the type of adsorption can be considered as ion-exchange mechanism. Determination of the thermodynamic parameters DeltaH(0), DeltaS(0) and DeltaG(0) showed that adsorption was spontaneous and exothermic in nature. It was also added that adsorption of TMP by KSF may involve physical adsorption. PMID- 16310308 TI - Performance optimization of coagulant/flocculant in the treatment of wastewater from a beverage industry. AB - This study investigated the effect of coagulation/flocculation treatment process on wastewater of Fumman Beverage Industry, Ibadan, Nigeria. The study also compared different dosages of coagulant, polyelectrolyte (non-ionic polyacrylamide) and different pH values of the coagulation processes. The effect of different dosages of polyelectrolyte in combination with coagulant was also studied. The results reveal that low pH values (3-8), enhance removal efficiency of the contaminants. Percentage removal of 78, 74 and 75 of COD, TSS and TP, respectively, were achieved by the addition of 500 mg/L Fe2(SO4)3.3H2O and 93, 94 and 96% removal of COD, TSS and TP, respectively, were achieved with the addition of 25 mg/L polyelectrolyte to the coagulation process. The volume of sludge produced, when coagulant was used solely, was higher compared to the use of polyelectrolyte combined with Fe2(SO4)3.3H2O. This may be as a result of non ionic nature of the polyelectrolyte; hence, it does not chemically react with solids of the wastewater. Coagulation/flocculation may be useful as a pre treatment process for beverage industrial wastewater prior to biological treatment. PMID- 16310309 TI - Haemonchus contortus and Trichostrongylus colubriformis in pen-trials with Javanese thin tail sheep and Kacang cross Etawah goats. AB - Weight gain costs due to infection were higher in sheep than goats, 28 and 17.5%, respectively, for Trichostrongylus colubriformis and 48.7 and 32.2%, respectively, for Haemonchus contortus. The extent of bodyweight cost attributed to anorexia in sheep infected with H. contortus was higher (13.5 g/day) than in sheep infected with T. colubriformis (2.3 g/day). On the other hand, bodyweight cost due to the other pathogenic effects in sheep infected with T. colubriformis were higher (35.6 g/day) compared to sheep infected with H. contortus (10.9 g/day). A strong relationship between faecal egg count and worm count (r=0.79, P=0.006) was shown only in sheep infected with T. colubriformis. About half of the infected sheep and goats had low or zero faecal egg counts throughout the study. In about 40% the egg count rose initially but became low by weeks 10-16, whereas in about 10% counts increased progressively throughout the period of observation and these animals also had the highest numbers of worms at slaughter. Packed cell volume was reduced in sheep and goats infected with H. contortus but serum protein and haemoglobin levels were unaffected. Sheep infected with T. colubriformis had a higher level of eosinophilia after 8 weeks (18.4%) than sheep infected with H. contortus (11.4%), whereas this pattern was reversed in goats and levels were also lower (4.1 and 8.9%, respectively). There was no apparent relationship between eosinophilia and resistance to infection with H. contortus or T. colubriformis. PMID- 16310310 TI - Quantitation of Leishmania lipophosphoglycan repeat units by capillary electrophoresis. AB - The glycosylphosphatidylinositol (GPI)-anchored lipophosphoglycan (LPG) of Leishmania is the dominant cell surface glycoconjugate of these pathogenic parasites. LPG is structurally characterized by a series of phosphoglycan repeat units. Determining the number of repeat units per LPG molecule has proven difficult using current technologies, such as mass spectrometry. As an alternative method to quantitate the number of repeat units in LPG, a procedure based on capillary electrophoretic analysis of the proportion of mannose to 2,5 anhydromannose (derived from the nonacetylated glucosamine of the GPI anchor of LPG) was developed. The CE-based technique is sensitive and relatively rapid compared to GC-MS-based protocols. Its application was demonstrated in quantitating the number of LPG repeat units from several species of Leishmania as well as from two life-cycle stages of these organisms. PMID- 16310311 TI - Effects of ovarian hormones on cognitive function in nonhuman primates. AB - Several studies have suggested that estrogen benefits verbal memory and lowers the risk of Alzheimer's disease in women, and improves cognitive function in animal models. However, the negative outcome of the Women's Health Initiative Memory Study has challenged the rationale for using estrogen as a protective agent against age-related cognitive decline. In view of the limitations of the Women's Health Initiative Memory Study, it is clear that our understanding of estrogen effects would greatly benefit from further interactions between clinical and basic science. Animal models of menopause can provide crucial information regarding the consequences of estrogen loss and replacement on several systems, including cognition. In this paper, I review the evidence that nonhuman primates, who share numerous cognitive and physiological characteristics with humans, can substantially contribute to our understanding of estrogen influences on the brain and cognition. Studies in young adult females suggest that some aspects of cognition fluctuate with the menstrual cycle, but that ovariectomy and estrogen replacement have only modest effects on cognitive function. In contrast, data in aged, naturally or surgically menopausal monkeys indicate that estrogen modulates a broad range of cognitive domains. Neurobiological data are consistent with the cognitive findings and demonstrate an array of morphological and physiological changes in brain areas important for cognition following ovariectomy and/or estrogen replacement. It is concluded that nonhuman primates, by providing a bridge between rodent and human data, constitute invaluable models to further our understanding of hormonal actions on the brain and cognition and to develop effective hormonal interventions against brain and cognitive aging. PMID- 16310312 TI - Neuroactive steroids attenuate oxytocin stress responses in late pregnancy. AB - In late pregnant rats neuroendocrine stress responses, expressed as increased oxytocin secretion and activation of the hypothalamo-pituitary-adrenal axis, are attenuated. These adaptations preserve the oxytocin store for parturition and prevent pre-term birth, and protect the fetuses from adverse programming by exposure to excess glucocorticoid. Mechanisms of adaptations for oxytocin neurones are reviewed, using challenge with systemic interleukin-1beta, simulating activation of immune signaling by infection, as a stressor of special relevance in pregnancy. In virgin rats, systemic interleukin-1beta stimulates the firing of oxytocin neurones, and hence oxytocin secretion, but interleukin-1beta has no effects in late pregnant rats. This lack of response is reversed by naloxone treatment just before interleukin-1beta administration, indicating endogenous opioid suppression of oxytocin responses in late pregnancy. This opioid presynaptically inhibits noradrenergic terminals impinging on oxytocin neurones. Finasteride pretreatment, inhibiting progesterone conversion to allopregnanolone, a positive GABA(A) receptor allosteric modifier, also restores an oxytocin response to interleukin-1beta. This finasteride effect is reversed by allopregnanolone treatment. In virgin rats allopregnanolone attenuates the oxytocin response to interleukin-1beta, which is exaggerated by naloxone. The effects of naloxone and finasteride in late pregnant rats in restoring an oxytocin response to interleukin-1beta are not additive. Accordingly, allopregnanolone may both enhance GABA inhibition of oxytocin neurone responses to interleukin-1beta, and induce opioid suppression of noradrenaline release onto oxytocin neurones. PMID- 16310313 TI - Steroid receptor coregulator diversity: what can it mean for the stressed brain? AB - Glucocorticoid hormones modulate brain function and as such are crucial for responding and adjusting to physical and psychological stressors. Their effects are mediated via mineralo- and glucocorticoid receptors, which in large measure act as transcription factors to modulate transcription of target genes, in a receptor-, cell-, and state-specific manner. The nature and magnitude of these transcriptional effects depend on the presence and activity of downstream proteins, such as steroid receptor coactivators and corepressors (together: coregulators), many of which are expressed in the brain. We address the role of coregulators for mineralo- and glucocorticoid receptor-mediated modulation of gene transcription. We first address evidence from cell lines for the importance of coregulator stoichiometry for steroid signaling. The in vivo importance of coregulators-when possible specifically for glucocorticoid signaling in the brain is discussed based on knockout mice, transient knockdown of steroid receptor coactivators, and distribution and regulation of coactivator expression in the brain. We conclude that for a better understanding of modulation of brain function by glucocorticoids, it is necessary to take into account the role of coregulators, and to assess their importance relative to changes in hormone levels and receptor expression. PMID- 16310314 TI - Hormonal influence on song structure and organization: the role of estrogen. AB - The development of song in songbirds is a complex phenomenon that involves memory and learning, sensorimotor integration, and neural and muscular maturation. Gonadal hormones are involved in each of these steps, as they influence the differentiation of the neural song system, the incorporation and survival of neurones, and the development of muscles used for song production. In young males the development of song, therefore, is closely linked to the secretion of testosterone by the testicles. Castration results in the development of incomplete or unstable songs, and hormone replacement leads to the development of crystallized or stable song. However, testosterone does not act solely as an androgen. The brain of songbirds contains high concentrations of the enzyme aromatase, which converts testosterone into estradiol. Estradiol then binds to estrogen receptors, which in the song system are found only in the nucleus HVC. This forebrain nucleus, also called the "master nucleus," codes for the syntactic structure of song, i.e. for the particular combination of simple elements syllables-that characterize the song of an individual. In this paper, we will review our studies on the role of estrogen in guiding the organization of song in canaries. PMID- 16310315 TI - Hippocampal synthesis of estrogens and androgens which are paracrine modulators of synaptic plasticity: synaptocrinology. AB - Hippocampal pyramidal neurons and granule neurons of adult male rats are equipped with a complete machinery for the synthesis of pregnenolone, dehydroepiandrosterone, testosterone, dihydrotestosterone and 17beta-estradiol. Both estrogens and androgens are synthesized in male hippocampus. These brain steroids are synthesized by cytochrome P450s (P450scc, P45017alpha and P450arom), hydroxysteroid dehydrogenases and reductases from endogenous cholesterol. The expression levels of enzymes are as low as 1/300-1/1000 of those in endocrine organs. Synthesis is dependent on the acute Ca(2+) influx upon neuron-neuron communication via NMDA receptors. Estradiol is particularly important because estradiol rapidly modulates neuronal synaptic transmission such as long-term potentiation via synaptic estrogen receptors. Xenoestrogens may also act via estrogen-driven signaling pathways. PMID- 16310316 TI - Cell death and sexual differentiation of the nervous system. AB - Sex differences in nuclear volume or neuron number often are attributed to the hormonal control of cell death. In the spinal nucleus of the bulbocavernosus, the central portion of the medial preoptic nucleus, and the principal nucleus of the bed nucleus of the stria terminalis testicular hormones decrease cell death during perinatal life, resulting in a male advantage in neuron number in adulthood. Conversely, males have more dying cells during development and fewer neurons in adulthood than do females in the anteroventral periventricular nucleus of the hypothalamus. This review discusses several limitations and unresolved issues in the literature on sexually dimorphic cell death, and identifies molecular mechanisms by which gonadal steroids may control cell survival. In particular, evidence is presented for the hormonal regulation of neurotrophic factors and involvement of Bcl-2 family proteins in the determination of sex differences in neuron number. PMID- 16310317 TI - Anabolic androgenic steroids and forebrain GABAergic transmission. AB - Anabolic androgenic steroids are synthetic derivatives of testosterone designed for therapeutic purposes, but now taken predominantly as drugs of abuse. The most common behavioral effects associated with anabolic androgenic steroid use are changes in anxiety, aggression and reproductive behaviors, including the onset of puberty and sexual receptivity. GABAergic circuits in the forebrain underlie these behaviors and are regulated by gonadal steroids. Work from our laboratories has shown that the expression and function of GABA(A) receptors in the rat and mouse forebrain varies between the sexes and across the estrous cycle. We have also shown that there are significant changes in GABA(A) receptor expression that occur with the progression through puberty to adulthood. Because GABAergic systems are both steroid-sensitive and critical for the expression of behaviors altered with anabolic androgenic steroid use, forebrain GABA(A) receptors are an attractive candidate to assess how molecular actions of anabolic androgenic steroids may be translated to known behavioral outcomes. Our studies demonstrate that anabolic androgenic steroids elicit both acute modulation of GABA(A) receptor-mediated currents, as well as chronic regulation of GABA(A) receptor expression and forebrain GABAergic transmission. Because anabolic androgenic steroid use has now become prevalent not only among adolescent boys, but in an increasing number of adolescent girls, we have also been particularly interested in determining age- and sex-specific effects of anabolic androgenic steroids. Our data show that the effects of chronic anabolic androgenic steroid exposure can be greater for adolescent than adult animals and are more marked in females than in males. These data have particularly important implications with respect to studies we have done demonstrating that chronic anabolic androgenic steroid exposure alters the onset of puberty, estrous cyclicity and sexual receptivity. PMID- 16310318 TI - The role of androgens in cognition and brain aging in men. AB - Losses of working and long-term memory are hallmarks of human aging and may signal impending neurodegenerative disease. The maintenance of neural elements in brain systems that support memory, such as synapse formation in prefrontal cortex and hippocampus, are critical for cognitive health in aging. This paper reviews the biological basis for androgens as neuroprotectants or neuromodulators in aging and the importance of androgens on the brain systems important for memory. We relate biological effects to cognitive outcomes in elderly men under a variety of androgen conditions. In brief, androgen deprivation causes significant loss of synapses in the hippocampus in rodent and nonhuman primates, increases amyloid deposition in human and rodent models and causes changes in neurotransmission in prefrontal cortex in rodent models. Recent work suggests that these changes modify age-related cognitive loss, particularly to memory in men. In addition, the conversion of testosterone to its androgen metabolites or to estradiol may play a special role in the preservation of memory in aging. This paper reviews discrepancies between studies using animal models and studies of human cognition, and suggests new directions that are likely to be fruitful in the future for understanding the role of androgens in brain aging. This review suggests that studies of low androgen levels in older men may not index the same biological mechanisms and behavioral effects as the studies of gonadectomy in animal models. PMID- 16310319 TI - Effects of gonadal hormones on central nitric oxide producing systems. AB - Nitric oxide-containing neurons are widely distributed within the CNS, including regions involved in the control of reproduction and sexual behavior. The expression of neuronal nitric oxide synthase is influenced by testosterone in male rat, and by estrogens in female. Moreover, nitric oxide synthase may co localize with gonadal hormones' receptors. Gonadal hormones may influence nitric oxide synthase expression in adulthood as well as during the development. In fact, in mice knockout for estrogen receptor alpha, the nitric oxide synthase expressing population is deeply reduced in specific regions. In physiological conditions, the female in mammalian species is exposed to short-term changes of gonadal hormones levels (estrous cycle). Our recent studies, performed in the rat vomeronasal system and in mouse hypothalamic and limbic systems reveal that, in rodents, the expression of nitric oxide synthase-producing elements within regions relevant for the control of sexual behavior is under the control of gonadal hormones. The expression of nitric oxide synthase may vary according to the rapid variations of hormonal levels that take place during the estrous cycle. This seems in accordance with the hypothesis that gonadal hormone activation of nitric oxide-cyclic guanosine-monophosphate pathway is important for lordosis behavior, as well as that this system is activated during mating behavior. Finally, comparative data available for other vertebrates suggest that class specific and species-specific differences occur in the nitric oxide synthase system of hypothalamus and limbic structures. Therefore, particular caution is needed to generalize data obtained from studies in rodents. PMID- 16310320 TI - Estrogen therapy: does it help or hurt the adult and aging brain? Insights derived from animal models. AB - Hormone therapy and estrogen therapy in postmenopausal women have been thought to ameliorate cognitive dysfunction and decrease the risk and/or progress of neurodegenerative conditions such as Alzheimer's disease and stroke. Furthermore, estrogens have been shown to exert neuroprotective actions in a variety of in vitro and in vivo models of brain injury. However, the findings of the Women's Health Initiative have made us re-evaluate these assumptions. Our laboratory has shown that physiological levels of estradiol attenuate ischemic brain injury in young and middle-aged female rats. We have begun to probe the cellular and molecular mechanisms that underlie these novel non-reproductive actions of this steroid. Our findings demonstrate that in both young and aging rats, treatment with physiological concentrations of estradiol decreases ischemic injury by almost 50%, compared with oil-treated controls. Additionally, our data suggest that estradiol acts by altering the expression of genes that suppress apoptosis and enhance survival in the penumbral region of the infarct. These observations demonstrate that estrogen therapy protects against stroke-related injury in young and aging female rats and strongly suggest that middle-aged animals remain responsive to the protective actions of estradiol. Furthermore, they suggest that estrogen therapy protects against cell death by influencing the expression of genes that suppress apoptotic cell death pathways. PMID- 16310322 TI - Non-fatal overdose and subsequent drug treatment among injection drug users. AB - Overdose is a leading cause of death among illicit drug users. Nine hundred twenty-four injection drug users (IDUs) in Baltimore, Maryland, were interviewed to characterize overdose events and determine the circumstances under which they lead to drug treatment. Overall, 366 (39.7%) reported at least one non-fatal drug overdose. Most (96.2%) used heroin on the day of their last overdose and almost half (42.6%) used heroin and alcohol but few (4.1%) used tranquilizers or benzodiazepines. Five percent were in drug treatment when the overdose occurred and 7.1% had been incarcerated 2 weeks prior. One in four IDUs (26.2%) sought drug treatment within 30 days after their last overdose of whom 75% enrolled. Speaking with someone about drug treatment after the overdose was associated with treatment seeking (AOR 5.22; 95% CI: 3.12, 8.71). Family members were the most commonly cited source of treatment information (53.7%) but only those who spoke with spouses, crisis counselors and hospital staff were more likely to seek treatment. Not being ready for treatment (69.6%) and not viewing drug use as a problem (30.7%) were the most common reasons for not seeking treatment and being placed on a waiting list was the most common reason for not subsequently enrolling in treatment (66.7%). Of the IDUs treated by emergency medical technicians, ER staff or hospital staff, only 17.3%, 26.2% and 43.2% reported getting drug treatment information from those sources, respectively. Interventions that provide drug treatment information and enhance motivation for treatment in the medical setting and policies that reduce barriers to treatment entry among motivated drug users are recommended. PMID- 16310321 TI - Sexual differentiation of central vasopressin and vasotocin systems in vertebrates: different mechanisms, similar endpoints. AB - Vasopressin neurons in the bed nucleus of the stria terminalis and amygdala and vasotocin neurons in homologous areas in non-mammalian vertebrates show some of the most consistently found neural sex differences, with males having more cells and denser projections than females. These projections have been implicated in social and reproductive behaviors but also in autonomic functions. The sex differences in these projections may cause as well as prevent sex differences in these functions. This paper discusses the anatomy, steroid dependency, and sexual differentiation of these neurons. Although the final steps in sexual differentiation of vasopressin/vasotocin expression may be similar across vertebrate species, what triggers differentiation may vary dramatically. For example, during development, estrogen masculinizes vasopressin expression in rats but feminizes its counterpart in Japanese quail. Apparently, nature consistently finds a way of maintaining sex differences in vasopressin and vasotocin pathways, suggesting that the function of these differences is important enough that it was conserved during evolution. PMID- 16310323 TI - From data to evidence, to action: findings from a systematic review of hospital screening studies for high risk alcohol consumption. AB - OBJECTIVE: To conduct a systematic review of hospital alcohol screening studies to identify effective and efficient evidence-based strategies. METHOD: Sixty-five studies (N=100,980) of alcohol problem prevalence amongst hospital patients were reviewed. RESULTS: Prevalence of positive alcohol screens varied according to hospital location, screening tool and patient characteristics. BAC measures (26%) were nearly twice as likely (OR=1.92, p<.001) to reveal positive screens in the ED than self-reports (16%). No difference was found in prevalence of self-report positive screens between ED (16%) and ward settings (17%). Males were two to four times more likely than females to screen positive (BAC: OR=2.37, p<.001, ED self report: OR=3.07, p<.001, ward self-report: OR=4.30, p<.001). ED patients aged 20 40 years and ward patients aged 30-50 years had the highest prevalence of positive screens. CONCLUSIONS: Prevalence of risky or problematic drinking among hospital patients is high and warrants systematic screening and intervention. Many hospitals lack sufficient resources to undertake widespread screening programs. For optimum return on resources, it is recommended to screen males in the ED using BAC measures. Established protocols applying priority criteria and staff training can increase screening accuracy and effectiveness. PMID- 16310324 TI - Variants of the 5'-untranslated region of human NCF2: expression and translational efficiency. AB - The NCF2 gene encodes p67(phox), an essential component of the multi-protein NADPH oxidase enzyme in phagocytic leukocytes, as well as in certain non phagocytic cells. In humans, the NCF2 gene is expressed as multiple NCF2 variants that differ in the 5'-untranslated region (5'-UTR). Previously, we reported the presence of four NCF2 5'-UTR mRNA variants (designated as NCF2 exon 1, intron 1a, intron 1b and intron 1c). As each of the gene variants encodes an identical p67(phox) protein, the functional significance of these message variants was not apparent. In this study, we investigated the relative expression levels and tissue-specificity of NCF2 5'-UTR variant mRNAs and their translation efficiency and stability. NCF2 5'-UTR variant transcripts were differentially expressed in various cell lines and human tissues. In vitro translation assays indicated that the NCF2 5'-UTR variants also differed in their effects on the translation of a luciferase reporter mRNA and NCF2 mRNA. Notably, NCF2 intron 1 5'-UTR variants, which are the predominantly expressed variants found in vivo, strongly inhibited translation when compared to the NCF2 exon 1 5'-UTR variant. In contrast, RNA decay assays demonstrated that there was no significant difference between stability of NCF2 intron 1 transcripts and the exon 1 5'-UTR variant in HL-60, MonoMac 6, and U937 cells. Moreover, expression of the variant transcripts remained unchanged after neutrophil phagocytosis, and was similar in normal neutrophils and neutrophils from a patient with X-linked chronic granulomatous disease. These studies suggest that expression of p67(phox) is regulated through mechanisms that include modulation of transcription and translation. PMID- 16310325 TI - SmpB: a novel outer membrane protein present in some Brachyspira hyodysenteriae strains. AB - A novel outer membrane protein-encoding gene was identified in Brachyspira hyodysenteriae. The predicted protein, SmpB, was encoded by a gene that contains regions of identity with that encoding the previously identified lipoprotein SmpA. However, the majority of the reading frame encoding SmpA and SmpB share no detectable similarity. Analysis of several strains revealed that B. hyodysenteriae harbours either smpA or the newly identified gene smpB, but not both. smpB encodes for a slightly larger protein than smpA, 17.6 and 16.8 kDa, respectively. The predicted proteins share an identical leader sequence and the first 10 amino acids of the mature protein, however, the remainder of the predicted protein sequence shows no similarity. It is hypothesised that smpA and smpB are present on the same area of the chromosome. The proteins are antigenically unique, as antisera raised against a strain of B. hyodysenteriae that expresses SmpA cannot detect SmpB and vice versa. Although the presence of an identical leader peptide suggests identical localisation of SmpA and SmpB, it is not known if the two predicted proteins share similar function. PMID- 16310326 TI - Macular pigmentary changes as a sequelae of retinal hemorrhages in premature infants with retinopathy of prematurity. AB - PURPOSE: We assess the incidence of macular changes in ROP patients with retinal hemorrhages. PATIENTS AND METHODS: The premature group consisted of 360 children born 32 weeks gestation and/or with weight below 1,500 g. We used the RetCam-120 Digital Retinal Camera to document retinal changes. RESULTS: Of the 360 premature infants 241 (67%) had no ROP, and 119 (33%) had ROP. Of the preterm infants with ROP retinal hemorrhages were found in 46 (38%) children. Of the newborns with ROP and with retinal hemorrhages, macular pigmentary changes were found in 3 (6%) patients. Of these 3 patients, the first had pre-threshold ROP, the second threshold ROP and had underwent diode laser photocoagulation, and the third patient had stage 2 ROP. In the patients with pre-threshold and threshold ROP retinal hemorrhages appeared 6 weeks after birth and macular pigmentary changes were found 6 months after birth. In the patient with stage 2 ROP hemorrhages appeared 7-8 weeks after birth and macular pigmentary changes were detected 12 months after birth. CONCLUSIONS: Although macular hemorrhages almost always resorb without complications, our study allows the assumption that retinal hemorrhages may cause macular pigmentary changes in the macula, and thus may lead to deprivation amblyopia. Our results may suggest that the presence of the macular pigmentary changes may be related to the hemorrhage and not to the specific therapy or to the disease. PMID- 16310327 TI - Spectrum of neural electrical activity in guinea pig cochlea: effects of anaesthesia regimen, body temperature and ambient noise. AB - Spectral analysis of electric noise recorded from the round window of the cochlea is thought to represent the summed spontaneous activity of the auditory nerve. It has been postulated that it could provide a possible tinnitus index. Because experimental conditions could change this neural activity, the effect of anaesthesia regimen, body temperature and ambient noise on the spectrum of spontaneous neural noise (SNN) were investigated in guinea pig cochlea. SNN was studied in awake guinea pigs and after anaesthesia with pentobarbital (P), xylazine/ketamine (XK) or xylazine/tiletamine-zolazepam (XTZ). Body temperature varied gradually from 33 to 41 degrees C under XK regimen. In awake animals, broadband noise was generated with intensity varying from 0 to 50 dB. The SNN consisted in a broad peak at approximately 900 Hz. With ambient broadband noise, it increased exponentially with the sound level with no shift in frequency. Soon after anaesthetic induction, the lowest frequencies were constantly decreased, and gradually the 900 Hz peak either increased moderately (P) or dropped steeply (XTZ) or remained unchanged (XK). Peak frequency increased linearly with body temperature whereas the amplitude reached a maximum at around 39.5 degrees C. In conclusion, these data indicate that experimental conditions such as anaesthesia regimen, body temperature and ambient noise modify the spontaneous neural outflow of the cochlea and must be taken into account when studying SNN. PMID- 16310328 TI - Suicide by carbon monoxide from car exhaust-gas in Denmark 1995-1999. AB - In the period 1995-1999 there were 388 car exhaust-gas suicides in Denmark. Of these 343 (88.4%) were men and 45 (11.6%) were women, the average age being 47 years. The car exhaust-gas suicides made up 9.3% of all suicides in Denmark in the period. The corresponding rate was 11.7% for men and 3.7% for women. In rural areas a larger part of all suicides were committed with car exhaust-gas compared to the more densely populated areas. Mental disease was diagnosed in 124 (32.0%) cases. A suicide note was found in 165 (42.5%) cases. A hose was fitted to the exhaust pipe in 334 (86.1%) cases. Of these the 234 (60.3%) occurred outside, typically in a forest area, while 76 (19.6%) occurred in a closed garage. All the 54 (13.9%) cases with no hose fitted to the exhaust pipe occurred in a garage. Seven (1.8%) victims were found in a burning or burnt-out car, where the following investigation revealed that it was actually a car exhaust-gas suicide. Carboxyhemoglobin was measured in 26 (6.7%) victims. In two of these victims no carboxyhemoglobin was found, as they had survived for some time after the poisoning. The average saturation of the remaining victims was 67%, the lowest saturation being 20% and the highest being 84%. In the period 1969-1987 the number of car exhaust-gas suicides in Denmark increased from 50 to approximately 190 per year and the rate of car exhaust-gas suicides compared to all suicides increased from approximately 5% to approximately 13%. In 1987-1999 these figures decreased from approximately 190 to 63 per year and from 13% to approximately 8%. During these 30 years the number of passenger cars in Denmark doubled, which explains the increase in car exhaust-gas suicides during 1969-1987. A possible explanation for the decrease in 1987-1999 is the introduction of the catalytic converter, which was made mandatory in 1990. We anticipate that car exhaust-gas suicides will continue to decrease in numbers, as more cars are equipped with catalytic converters. PMID- 16310329 TI - [Chemosusceptibility analysis of Plasmodium falciparum imported from Comoros to Marseilles, France in 2001-2003]. AB - OBJECTIVES: The aim of this work was to study the chemosensitivity of Plasmodium falciparum strains isolated from patients presenting with malaria after having returned from Comoros Islands in 2002-2003, and hospitalized at the North University Hospital, in Marseilles, France. MATERIALS AND METHODS: In vitro drug susceptibility (for strains maintained in culture) and mutation-specific polymerase chain reaction (PCR) assays (for all strains) were performed. RESULTS: Out of 23 strains kept in culture, 50% were shown to be resistant in vitro to chloroquine, 50% were resistant to pyrimethamine, 40% to cycloguanil, 25% to atovaquone, and 7% to mefloquine. However all these strains were susceptible to quinine, halofantrine, and artemether. Moreover, 48 strains were tested by molecular methods. As a result, 69% were shown to have the Asp108 mutation in the dihydrofolate reductase gene (Pfdhfr), the basic mutation associated with antifolate resistance, and 54% had additional mutations Ile51 plus Arg59, associated with a high level of resistance. Furthermore, 90% of the 20 strains tested in 2003 were shown to have the point mutation Pfcrt76 in the P. falciparum chloroquine resistance transporter (Pfcrt) gene recently proposed as a molecular marker of chloroquine-resistance. CONCLUSION: Obtaining plasmodium strains from Comoros to be tested in Marseilles, where all laboratory facilities are available, is a unique opportunity to establish a surveillance of falciparum drug resistance in the Comoros islands. PMID- 16310330 TI - Differential stimulation of muscle protein synthesis in elderly humans following isocaloric ingestion of amino acids or whey protein. AB - To counteract the debilitating progression of sarcopenia, a protein supplement should provide an energetically efficient anabolic stimulus. We quantified net muscle protein synthesis in healthy elderly individuals (65-79 yrs) following ingestion of an isocaloric intact whey protein supplement (WY; n=8) or an essential amino acid supplement (EAA; n=7). Femoral arterio-venous blood samples and vastus lateralis muscle biopsy samples were obtained during a primed, constant infusion of L-[ring-2H5]phenylalanine. Net phenylalanine uptake and mixed muscle fractional synthetic rate (FSR) were calculated during the post absorptive period and for 3.5 h following ingestion of 15 g EAA or 15 g whey. After accounting for the residual increase in the intracellular phenylalanine pool, net post-prandial phenylalanine uptake was 53.4+/-9.7 mg phe leg-1 (EAA) and 21.7+/-4.6 mg phe leg-1 (WY), (P<0.05). Postabsorptive FSR values were 0.056+/-0.004% h-1 (EAA) and 0.049+/-0.006% h-1 (WY), (P>0.05). Both supplements stimulated FSR (P<0.05), but the increase was greatest in the EAA group with values of 0.088+/-0.011% h-1 (EAA) and 0.066+/-0.004% h-1 (WY), (P<0.05). While both EAA and WY supplements stimulated muscle protein synthesis, EAAs may provide a more energetically efficient nutritional supplement for elderly individuals. PMID- 16310331 TI - Does infertility cause marital benefit? An epidemiological study of 2250 women and men in fertility treatment. AB - OBJECTIVE: To investigate (i) marital benefit, e.g., that infertility has strengthen the marriage and brought the partners closer together among people beginning fertility treatment and (ii) communication and coping strategies as predictors of marital benefit 12 months later. METHODS: A prospective cohort design including 2250 people beginning fertility treatment and a 12-month follow up. Data were based on self-administered questionnaires measuring marital benefit, communication, and coping strategies. The analyses of predictors were based on the sub-cohort (n=816) who had not achieved a delivery after fertility treatment. RESULTS: 25.9% of women and 21.1% of men reported high marital benefit. Among men medium use of active-confronting coping (e.g., letting feelings out, asking others for advice) and use of meaning-based coping were significant predictors for high marital benefit. Having the infertility as a secret, difficult marital communication, and using active-avoidance coping (e.g., avoid being with pregnant women or children, turning to work to take mind off things) were among men significant predictors for low marital benefit. No significant predictors were identified among women. CONCLUSION: Fertility patients frequently experience marital benefit. PRACTICE IMPLICATIONS: The study provides information about where to intervene with male fertility patients in order to increase their marital benefit after medically unsuccessful treatment. PMID- 16310332 TI - [Cardiovascular risk and chronic kidney failure before kidney transplantation. Is coronary angiography essential?]. PMID- 16310333 TI - [Complication of tracheal intubation: severe cervical cellulitis]. AB - A 40-year-old man, victim of a traffic accident has been hospitalized for a severe head trauma. His trachea has been intubated under general anaesthesia with an 8.0 mm ID tube (Vygon). The cerebral scan revealed a surgical subdural haematoma. In the postoperative period, the patient was admitted in surgical intensive care, under sedation and mechanically ventilation. At day 12 the patient developed a cervical cellulitis complicated of a septic shock. The cervical scan showed an important dilatation of the trachea in the site of the tube cuff. The surgical exploration discovered a complete destruction of the anterior face of de trachea on several centimetres of height. The patient died 24 hours later by multiple organ failure. PMID- 16310334 TI - [Diagnosis and management of a phaeochromocytoma discovered with laparoroscopic CO2 insufflation]. PMID- 16310335 TI - [Anaesthetic particularities for children with tumours]. AB - OBJECTIVES: To present the more frequent solid tumors, that require a general anaesthesia and to describe the particularities of the anaesthetic management. DATA SOURCES AND EXTRACTION: The PUBMED database was searched for articles (1990 2005) combined with references analysis of major articles on the topics. DATA SYNTHESIS: Neoplasma is the first cause of paediatric death. In children less than 5 years of age, neuroblastoma and nephroblastoma are the more frequent of the tumour. The incidence of each pathology is low. However a general anaesthesia is required in numerous situations: bone marrow aspiration and biopsy, central venous catheter insertion, various biopsies under computed tomography, ultrasound, thoracoscopy or laparoscopy, MRI scan, surgery and body radiation. The key points of successful anaesthetic management include complete preoperative evaluation together with the oncology paediatric team. Particularly, cardiotoxicity may result from chemotherapy and hypertension is frequently associated with neuroblastoma and nephroblastoma. Mediastinal location of lymphomas induced a significant risk of cardiopulmonary complications during induction of general anaesthesia. A peak expiratory flow rate or a tracheal cross section area less than 50% of predicted values are contraindication to general anaesthesia. Cephalic rhabdomyosarcoma may be responsible of difficult airway management. Specific guidelines are available for platelet and red cell transfusion in paediatric oncology patients. Surgical antibioprophylaxis are adapted in immunodepressed children to a potential increased risk of postoperative infectious complications, according personal previous history, microbiological ecology and body bacterial flora. Dosage of anaesthetic agents takes into account denutrition, hydroelectrolytic disturbances and hypovolemia. PMID- 16310336 TI - [Preoperative information for paediatric patients. The anaesthesiologist's point of view]. AB - Medical information is mandatory before any medical procedure, including pediatric anesthesia. Preoperative information covers many aspects, including medico-psychologic and judicial aspects. When the patient is a child, information must be delivered to the parents in priority. However, the French law has given a particular attention to the opinion of the child. In 70% of the cases, preoperative anxiety of the parents is more related to anesthesia than to the surgical procedure itself. We have to explain the most frequent adverse effects, as well as the more severe and well known complications to the parents, even though they are very infrequent. The only cases where preoperative information is not required are: emergency cases and refusal of the patient or the parents to be informed. While information is necessarily oral, it may be completed using a written document. The quality of the preoperative information directly influences the quality of the psychological preoperative preparation of the parents, and thereafter of the child. Preoperative preparation programs have been developed, but controversial results have been observed. The great majority of the studies on preoperative programs were performed in the USA, where the demand for preoperative information is very important. It is far from sure that the results of all these studies may be extrapolated to French parents, and French studies are needed. PMID- 16310337 TI - [Epidural blood patch, never too late]. AB - Long lasting postural puncture headache secondary to an unintentional dural puncture (UDP) during epidural anaesthesia (EA) or following deliberate dural puncture for spinal anaesthesia, and neurological or neuroradiological examination, is becoming unusual. Placing the bevel of Tuohy needle parallel to the longitudinal axis of the dural cylinder when searching for epidural space, and in the later cases, widespread use of small diameter, atraumatic needles, have decreased both its incidence and severity. Furthermore, in the absence of any contraindication, patients suffering from such troubles are now quite systematically offered a curative epidural blood patch (EBP) whose effectiveness is documented. Authors report an original case of a fainthearted patient who underwent caesarean section under EA when she was 31-year-old, and whose disabling painful symptoms related to a UDP were successfully relieved by one EBP that she ended to accept more than seven years later. On the occasion of a short literature review, physiopathology and management are debated. PMID- 16310338 TI - [Caesarean section in a parturient with C1 esterase inhibitor deficiency (angioneurotic oedema)]. PMID- 16310339 TI - [What are the changes in paediatric anaesthesia practice afforded by new anaesthetic ventilators?]. AB - Because of specific paediatric respiratory physiology (mainly decreased compliance and functional residual capacity, increased O2 demand and CO2 production), ventilators for paediatric anaesthesia need to be powerful and able to deliver small volumes at a high rate without compression volume loss. The compensation of compliance now available on every anaesthesia machine, compensates for the volume of gas lost by compression in the circuit tubing allowing the tidal volume to reach preset commands, even for bellow in box respirators. Preset tidal volume is then totally delivered to the lung by volume controlled ventilation because it becomes independent of total pulmonary compliance and fresh gas flow. Increased precision of electronic flowmeters and better air-tightness of circuits allow reducing with precision fresh gas flow to values approaching children's O2 consumption and N2O diffusion. New modes of ventilation are now available on anaesthesia machine. Pressure controlled mode, by increasing and maintaining mean airway pressures, ameliorates intrapulmonary gas distribution and compensates for the gas leak from uncuffed tracheal tubes. Unsteady tidal volume resulting from variation of total compliance, is the main drawback of pressure-controlled ventilation that may be overcome by using the "autoflow" mode (better described as a pressure controlled mode ensuring tidal volume) available with one of the last generation of ventilators. Increased accuracy and security of the mode "pressure assist" might increase the use of spontaneous ventilation in paediatric anaesthesia even for low weight children. However tidal volume remains variable with compliance and depth of anaesthesia, which may require several adjustments of ventilator's settings. The clinical conditions (mainly airway control) of pressure assist use for children less than 10 kg should be elucidated before recommending its use. PMID- 16310340 TI - DNA reassociation using oscillating phenol emulsions. AB - Reassociating double-stranded DNA from single-stranded components is necessary for many molecular genetics experiments. The choice of a DNA reassociation method is dictated by the complexity of the starting material. Reassociation of simple oligomers needs only slow cooling in an aqueous environment, whereas reannealing the many single-stranded DNAs of complex genomic mixtures requires both a phenol emulsion to accelerate DNA reassociation and dedicated equipment to maintain the emulsion. We present a method that is equally suitable for reassociating either simple or complex DNA mixtures. The Oscillating Phenol Emulsion Reassociation Technique (OsPERT) was primarily developed to prepare heteroduplex DNA from alkali-denatured high molecular weight human genomic DNA samples in which hundreds of thousands of fragments need to be reannealed, but the simplicity of the technique makes it practical for less demanding DNA reassociation applications. PMID- 16310341 TI - Identification and functional characterization of a novel interleukin 17 receptor: a possible mitogenic activation through ras/mitogen-activated protein kinase signaling pathway. AB - Interleukin-17 receptor (IL-17R) is increasingly emerged as a distinct receptor family functioning in diverse cellular processes including inflammation and cancer. In this study, we uncovered a novel member of IL-17R from mouse tissue that was named mouse IL-17RE (mIL-17R). Mouse IL-17RE cDNA is composed of at least 14 exons and presents at least 6 spliced isoforms (mIL-17RE1-6) with a molecular weight ranging from 34.2 to 70.1 kD. Mouse IL-17RE is expressed in limited tissues such as lung, kidney, stomach, intestine and testis, etc., and is mainly localized in the cytoplasm and on cell membrane. IL-17RE can also be detected in numerous tumor cell lines. Importantly, a mitogenic effect was detected in BaF3 cells stably transfected with the chimeric receptor fused by the ectodomain of erythropoietin receptor (EPOR) with the transmembrane and endomain of IL-17RE in a serum-dependent but EPO-independent manner. Moreover, ERK1/2 phosphorylation was significantly up-regulated as the dose of mIL-17RE increased. Specific RNAi targeting at mIL-17RE dramatically inhibited the activation of ERK1/2, indicating that mIL-17RE could functionally activate RAS/MAPK signaling pathway. Using dominant negative MEK (Dn-MEK) or RAS (Dn-RAS) as a signaling blocker, we were able to show that mIL-17RE probably activated RAS/MAPK signaling at or upstream of RAS. Overall, our results strongly indicate that mIL-17RE may belong to a novel growth-receptor like molecule that has the capability to support cellular mitogenesis through RAS/MAPK pathway. PMID- 16310342 TI - Phosphatidylinositol 3-kinase/Akt pathway is involved in transforming growth factor-beta1-induced phenotypic modulation of 10T1/2 cells to smooth muscle cells. AB - Transforming growth factor-beta1 (TGF-beta1) is known to induce phenotypic modulation of mesenchymal cells to SMCs. However, the intracellular signals regulating induction of the SMC phenotype of mesenchymal cells have not been fully clarified. In the present study, we examined the role of the mitogen activated protein kinase (MAPK) superfamily and phosphatidylinositol 3-kinase (PI3K)/Akt in the TGF-beta1-mediated phenotypic modulation of 10T1/2 mesenchymal cells to SMCs characterized by the expression of SMC-specific markers, including smooth muscle alpha-actin (SMalpha-actin), myosin heavy chain (SM-MHC), and protein 22-alpha (SM22alpha). The results showed the following: (1) TGF-beta1 induced SMalpha-actin and SM-MHC expressions in 10T1/2 cells in a time-dependent manner. (2) TGF-beta1 induced biphasic increases in extracellular signal regulated kinase (ERK), p38 MAPK, c-Jun-NH2-terminal kinase (JNK), and Akt phosphorylation. (3) The inhibitor for PI3K/Akt (i.e., LY294002), but not those for MAPKs (i.e., SB203580, PD98059, and SP600125), attenuated the TGF-beta1 induced SMalpha-actin and SM-MHC expressions in 10T1/2 cells; in addition, transfection of 10T1/2 cells with the Akt-specific small interfering RNA (siRNA) significantly reduced their SMalpha-actin and SM-MHC expressions. (4) LY294002 and the Akt-specific siRNA inhibited the TGF-beta1-induced SM22alpha gene expression and promoter activity, suggesting that the TGF-beta1-induced gene expression was mediated by PI3K/Akt at the transcriptional level. (5) LY294002 inhibited the TGF-beta1-induced gene expression and DNA binding activity of serum response factor (SRF). These results indicate that TGF-beta1 is capable of inducing the SMC phenotype of 10T1/2 cells and that this induction is mediated through the PI3K/Akt signaling pathway. PMID- 16310343 TI - Do you see what I see? Interpretations of intentional movement in schizophrenia. AB - OBJECTIVE: Current literature exploring theory of mind (ToM) abilities in patients with schizophrenia has failed to take into account the dynamic nature of complex social interactions. The aim of this study was to explore symptom specific impairments in theory of mind using a novel, dynamic task. METHODS: Subjects viewed short animations displaying three types of movement; random, goal directed, and socially complex (theory of mind). Verbal descriptions of the animations were obtained from 61 patients with schizophrenia (divided into symptom sub-groups) and 22 healthy comparison subjects and were scored for accuracy, type of response and use of target terms (terms most appropriate to each animation type). RESULTS: Accuracy on all three conditions discriminated behavioural signs (BS), and (to a lesser degree) paranoid subjects, from the other schizophrenia sub-groups (those in remission and those with passivity features) and the controls. Paranoid and BS groups had difficulties with all the animations, yet all symptom sub-groups failed to use the appropriate mentalising language to describe the ToM animations. CONCLUSIONS: In this first exploration of on-line mentalising abilities in schizophrenia, it is suggested that a failure to use appropriate mentalising language may be a trait marker for the disease. The nature of the type of tasks used to assess social cognitive processing in this group needs careful consideration, and tasks tapping into the fluidity of social interactions yield results that differ from previously reported studies. PMID- 16310344 TI - Phylogenetic analysis of members of the metabolically diverse genus Gordonia based on proteins encoding the gyrB gene. AB - Members of the metabolically diverse genus Gordonia were isolated from various biotopes including pristine and polluted sites around Taiwan. Identification, comparison and diversity assessment based on the gyrB gene were carried out using a newly developed primer pair for gyrB. The 16S rRNA gene was also sequenced for comparison. A 1.2-kb fragment of the gyrB gene of 17 Gordonia strains including type strains was determined by direct sequencing of PCR amplified fragments. A total of 25 strains (8 of which were retrieved from a public database) of the genus Gordonia form a distinct phyletic line in the GyrB-based tree and are separated from other closely related species of genera of the suborder Corynebacterineae. Sequence similarity of the gyrB sequence from twelve Gordonia type strains ranged from 79.3 to 97.2%, corresponding to between 270 and 41 nucleotide differences, while there was only a 0.3-3.8% difference in 16S rRNA gene sequence similarity at the interspecies level. Phylogenetic analysis based on the GyrB sequence deduced from the gyrB gene is consistent with that of DNA DNA hybridization results and provides a better discrimination within the species of Gordonia compared to the 16S rRNA gene. The present study demonstrates that gyrB gene analysis will aid in describing novel species belonging to the genus Gordonia. PMID- 16310345 TI - Assessing the quantitative reliability of solid-state 13C NMR spectra of kerogens across a gradient of thermal maturity. AB - Five type II kerogens, shown by elemental analysis and Rock-Eval pyrolysis to represent a gradient of thermal maturity, were further characterized using a range of solid-state 13C NMR spectroscopic techniques. 13C cross polarization (CP) NMR spectra of the kerogens confirmed the well-established pattern of increasing aromaticity with increasing thermal maturity. Spin counting showed that CP observability was around 50% for the immature kerogens, and only 14-25% for the mature kerogens. Spin counting also showed that the direct polarization (DP) observabilities were >80% for all but one of the kerogens. Despite the large differences in observability between the two techniques, aromaticities derived from corresponding CP and DP spectra differed by only 1-15%. The RESTORE technique showed that the low CP observability of the immature kerogens was due mostly to rapid T(1rho)H relaxation, whereas both rapid T(1rho)H relaxation and slow polarization transfer contributed to the low CP observability of the mature kerogens. PMID- 16310346 TI - The role of the left Brodmann's areas 44 and 45 in reading words and pseudowords. AB - In this functional magnetic resonance imaging (fMRI) study, we investigated the influence of two task (lexical decision, LDT; phonological decision, PDT) on activation in Broca's region (left Brodmann's areas [BA] 44 and 45) during the processing of visually presented words and pseudowords. Reaction times were longer for pseudowords than words in LDT but did not differ in PDT. By combining the fMRI data with cytoarchitectonic anatomical probability maps, we demonstrated that the left BA 44 and BA 45 were stronger activated for pseudowords than for words. Separate analyses for LDT and PDT revealed that the left BA 44 was activated in both tasks, whereas left BA 45 was only involved in LDT. The results are interpreted within a dual-route model of reading with the left BA 44 supporting grapheme-to-phoneme conversion and the left BA 45 being related to explicit lexical search. PMID- 16310347 TI - Utilization of the human cell line HL-60 for chemiluminescence based detection of microorganisms and related substances. AB - In this paper we describe a new pyrogen assay using the human leukemia cell line HL-60. The cell line is differentiated using all-trans retinoic acid (ATRA) to generate a cell population that resembles mature granulocytes. The differentiated HL-60 cell is capable of generating reactive oxygen species (ROS) when challenged with pyrogenic substances. In a luminol enhanced chemilumimetric assay the responsiveness of differentiated HL-60 cells is tested towards Salmonella typhimurium, Bacillus subtilis, Saccharomyces cerevisiae, Candida albicans, lipopolysaccharide (LPS) and lipoteichoic acid (LTA). The results show a poor sensitivity to S. typhimurium but displays good sensitivity towards B. subtilis, LTA and LPS. Furthermore, the sensitivity towards the yeasts C. albicans and S. cerevisiae is considerably better than obtained in other in vitro cell systems. Overall these results indicate that the HL-60 cell assay possibly could be evolved to a supplementary assay for the known pyrogenic detection assays. Furthermore, the utilization of the assay for pyrogenic examination of recombinant drugs derived from yeast expression systems would be relevant to examine. PMID- 16310348 TI - Feature detection using spikes: the greedy approach. AB - A goal of low-level neural processes is to build an efficient code extracting the relevant information from the sensory input. It is believed that this is implemented in cortical areas by elementary inferential computations dynamically extracting the most likely parameters corresponding to the sensory signal. We explore here a neuro-mimetic feed-forward model of the primary visual area (VI) solving this problem in the case where the signal may be described by a robust linear generative model. This model uses an over-complete dictionary of primitives which provides a distributed probabilistic representation of input features. Relying on an efficiency criterion, we derive an algorithm as an approximate solution which uses incremental greedy inference processes. This algorithm is similar to 'Matching Pursuit' and mimics the parallel architecture of neural computations. We propose here a simple implementation using a network of spiking integrate-and-fire neurons which communicate using lateral interactions. Numerical simulations show that this Sparse Spike Coding strategy provides an efficient model for representing visual data from a set of natural images. Even though it is simplistic, this transformation of spatial data into a spatio-temporal pattern of binary events provides an accurate description of some complex neural patterns observed in the spiking activity of biological neural networks. PMID- 16310349 TI - Comparing information about arm movement direction in single channels of local and epicortical field potentials from monkey and human motor cortex. AB - Cortical field potentials have been used for decades in neurophysiological studies to probe spatio-temporal activity patterns of local populations of neurons. Recently, however, interest in these signals was spurred as they were proposed as potential control signals for neuronal motor prostheses, i.e., for devices fit to record and decode brain activity to restore motor functions in paralyzed patients. Little is known, however, about the functional significance of these cortical field potentials. Here we compared information about arm movement direction in two types of movement related cortical field potentials, obtained during a four direction center-out arm reaching paradigm: local field potentials (LFPs) recorded with intracortical micro-electrodes from monkey motor cortex, and epicortical field potentials (EFPs) recorded with macro-electrode arrays subdurally implanted on the surface of the human cerebral cortex. While monkey LFPs showed a typical sequence of positive and negative potential peaks, an initial negative peak was the most salient feature of human EFPs. Individual contralateral LFPs from the monkey motor cortex carried approximately twice as much decoded information (DI) about arm movement direction (median 0.27 bit) as did individual EFPs from the contralateral hand/arm area of primary motor cortex in humans (median 0.12 bit). This relation was similar to the relation between median peak signal-to-noise ratios for directional modulation of movement related potentials (MRPs) of both types of signals. We discuss possible reasons for the observed differences, amongst them epi- vs. intracortical recording and the different electrode dimensions used to measure EFPs and LFPs. PMID- 16310350 TI - Fading memory and kernel properties of generic cortical microcircuit models. AB - It is quite difficult to construct circuits of spiking neurons that can carry out complex computational tasks. On the other hand even randomly connected circuits of spiking neurons can in principle be used for complex computational tasks such as time-warp invariant speech recognition. This is possible because such circuits have an inherent tendency to integrate incoming information in such a way that simple linear readouts can be trained to transform the current circuit activity into the target output for a very large number of computational tasks. Consequently we propose to analyze circuits of spiking neurons in terms of their roles as analog fading memory and non-linear kernels, rather than as implementations of specific computational operations and algorithms. This article is a sequel to [W. Maass, T. Natschlager, H. Markram, Real-time computing without stable states: a new framework for neural computation based on perturbations, Neural Comput. 14 (11) (2002) 2531-2560, Online available as #130 from: ], and contains new results about the performance of generic neural microcircuit models for the recognition of speech that is subject to linear and non-linear time warps, as well as for computations on time-varying firing rates. These computations rely, apart from general properties of generic neural microcircuit models, just on capabilities of simple linear readouts trained by linear regression. This article also provides detailed data on the fading memory property of generic neural microcircuit models, and a quick review of other new results on the computational power of such circuits of spiking neurons. PMID- 16310351 TI - [What is new in pediatric nephrology?]. AB - Recent advances in the knowledge of physiology, genetics, imaging, and therapeutics have lead to novel practical approaches in paediatric nephrology. Many inherited syndromes have been revisited in order to identify precise renal diseases at the molecular level. In addition, a large number of epidemiological studies and clinical trials have allowed guidelines and recommendations to be provided for chronic and end-stage renal failure, urinary tract infection, glomerular diseases, etc. PMID- 16310352 TI - Titration of non-invasive positive pressure ventilation in chronic respiratory failure. AB - Non-invasive ventilation (NIV) is widely used for acute and chronic respiratory failure. If arterial blood gas tensions do not improve, the level of support can be increased. However, there may be a limit above which increasing ventilatory support leads only to greater interface leak with no improvement in ventilation. The aim of this study was to establish whether there is such a limit. During a daytime study in 24 ventilated stable patients (10 with chronic obstructive pulmonary disease (COPD), 14 with chest wall deformity, CWD), inspiratory pressures up to 20 cm H(2)O and set tidal volumes up to 10 ml kg(-1) were associated with mask leak of <5 l min(-1). Although leak increased with higher levels of support, there was still an increase in minute ventilation. The mean (2 sd) tolerated pressure was 24 cm H(2)O (8-40) in both groups, and set tidal volume 12.7 ml kg(-1) (5.0-20.4) in CWD and 9.6 ml kg(-1) (3.9-14.8) in COPD. Measures of respiratory effort were significantly reduced at all levels with both forms of ventilatory support. There is debate about whether the therapeutic aim of NIV should be to reduce respiratory muscle effort, or to reverse nocturnal hypoventilation. We conclude that if the primary aim is to improve arterial blood gas tensions and this is not achieved, higher levels of ventilation can be obtained using greater pressure or volume, despite additional interface leak. If the aim is to abolish muscle effort completely, there is little to be gained by increasing the level of inspiratory pressure above 20 (CWD) or 25 (COPD) cm H(2)O. PMID- 16310353 TI - A strategy for sensitivity and specificity enhancements in prostate specific antigen-alpha1-antichymotrypsin detection based on surface plasmon resonance. AB - A biochip based on surface plasmon resonance was fabricated to detect prostate specific antigen-alpha(1)-antichymotrypsin (PSA-ACT complex) in both HBS buffer and human serum. To reduce non-specific binding and steric hindrance effect, the chemical surface of the sensor chips was constructed by using various oligo(ethylene glycol) mixtures of different molar ratios of HS(CH2)11(OCH2CH2)6OCH2COOH and HS(CH2)11(OCH2CH2)3OH. The self-assembled monolayers were biotinylated to facilitate the immobilization of streptavidin. Using the chip surfaces, PSA-ACT complex in HBS buffer and human serum was detected at 20.7 and 47.5 ng/ml by primary immunoresponse, respectively. However, the limit of detection could be simply enhanced by a sandwich strategy to improve the sensitivity and specificity of the immunoassay. An intact PSA polyclonal antibody was used as an amplifying agent in the strategy. As a result, PSA-ACT complex concentrations as low as 10.2 and 18.1 ng/ml were found in the HBS buffer and human serum sample, respectively. The result indicates that this approach could satisfy our goal without modifying the secondary interactant. PMID- 16310354 TI - The role of carbonic anhydrase IX overexpression in kidney cancer. AB - Carbonic anhydrase IX (CA IX) is a membrane isoenzyme, the overexpression of which is associated with clear cell carcinoma of the kidney. Its overexpression is restricted mainly to cancer, as it is absent in corresponding normal tissues making it a potential cancer biomarker. Several recent studies have shown that CA IX, apart from its classical enzyme activity of reversibly hydrating carbon dioxide extracellularly to facilitate the net extrusion of protons from inside to outside the cell, it can also be a key player in the modulation of cell adhesion processes and participate in the regulation of cell proliferation in response to hypoxic environment to ultimately contribute to tumour progression. Here, we have shown that the sole tyrosine moiety of CA IX present in its intracellular domain can be phosphorylated in an epidermal growth factor dependent manner, suggesting that it can feed into the growth factor receptor dependent signalling pathways. Our studies suggest that the tyrosine phosphorylated CA IX can interact with the regulatory subunit of PI-3-Kinase, contributing to Akt activation. These studies have revealed a positive feed back loop that can form the basis of a vicious cycle that could contribute to the progression of clear cell renal carcinoma and poor prognosis. These studies show that CA IX signalling may be a part of both the hypoxia driven and hypoxia independent pathways that occur in the cancer cell. Finally, our studies emphasize the need for a more refined strategy using signal transduction therapeutics to inhibit the cell surface carbonic anhydrases for the management of this malignancy. PMID- 16310355 TI - Inactivation of E-cadherin and less sensitivity of lobular breast carcinoma cells to chemotherapy. PMID- 16310356 TI - Reduction mammaplasty for benign phyllodes tumour in an adolescent female--a 13 year follow up. AB - Phyllodes tumours are rare neoplasms of the female breast, especially among adolescent women. We present the case of a 17-year old female patient with a large benign phyllodes tumour. The patient was treated by one-stage excision with reduction mammaplasty. Symmetry of breast contour and nipple-areola complex position could be restored. At follow-up after 13 years good cosmetic result was maintained and there was no evidence of local recurrence. Standard plastic surgical techniques like reduction mammaplasty might be applied in treating large benign tumours compromising the aesthetic aspect of the breast. PMID- 16310357 TI - The latest phospholipase C, PLCeta, is implicated in neuronal function. AB - Members of the phosphoinositide-specific phospholipase C (PLC) family have key roles in cell signalling. In response to many extracellular stimuli, such as hormones, neurotransmitters, antigens and growth factors, PLCs catalyse the hydrolysis of phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P(2)], thereby generating two well-established second messengers, inositol (1,4,5)-trisphosphate and diacylglycerol. Eleven PLC isozymes encoded by different genes have been identified in mammals and, on the basis of their structure and sequence relationships, have been classified into five families designated PLCbeta (1-4), PLCgamma (1 and 2), PLCdelta (1, 3 and 4), PLCepsilon (1) and PLCzeta (1). All PLCs contain the catalytic X and Y domain, in addition to other regulatory domains including the C2 domain and the EF-hand domain. In 2005, four groups independently identified an entirely new family of PLCs--eta1 and eta2--using data mining of mammalian genomes. The properties of the PLCeta enzyme suggest that it might act as a Ca(2+) sensor, in particular, functioning during formation and maintenance of the neuronal network in the postnatal brain. PMID- 16310358 TI - Synthesis, characterization, cytotoxic activities, and DNA-binding properties of the La(III) complex with Naringenin Schiff-base. AB - A new Naringenin Schiff-base ligand (H3L) and its complex, [La(H2L)2(NO3).3H2O], have been synthesized and characterized on the basis of elemental analyses, molar conductivities, mass spectra, 1H NMR, thermogravimetry/differential thermal analysis (TG-DTA), UV spectra, and IR spectra. Spectrometric titrations, ethidium bromide displacement experiments, and viscosity measurements indicate that the two compounds, especially the La(III) complex, strongly bind with calf-thymus DNA, presumably via an intercalation mechanism. The intrinsic binding constants of the La(III) complex and ligand with DNA were 1.83 x 10(7) and 9.46 x 10(5) M( 1), respectively. Comparative cytotoxic activities of the La(III) complex and ligand were also determined by MTT [3-(4,5-dimethyl-2-thiazoyl)-2,5-diphenyl-2H tetrazolium bromide] and SRB (sulforhodamine B) methods. The results showed that the La(III) complex had significant cytotoxic activity against the tested cells. PMID- 16310359 TI - New 1,3,4-thiadiazole derivatives endowed with analgesic and anti-inflammatory activities. AB - Two series of N-[5-oxo-4-(arylsulfonyl)-4,5-dihydro-1,3,4-thiadiazol-2-yl]-amides were synthesized and tested in vivo for their analgesic and anti-inflammatory activities. All the new compounds possess good antalgic action in the acetic acid writhing test and some terms of the series showed also fair anti-inflammatory activity in the carrageenan rat paw edema test. Ulcerogenic and irritative action on the gastrointestinal mucose, in comparison with indomethacin is low. PMID- 16310360 TI - Evaluation of fosmidomycin analogs as inhibitors of the Synechocystis sp. PCC6803 1-deoxy-D-xylulose 5-phosphate reductoisomerase. AB - Analogs of the antibiotic fosmidomycin, an inhibitor of the methylerythritol phosphate pathway to isoprenoids, were synthesized and evaluated against the recombinant Synechocystis sp. PCC6803 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR). Fosfoxacin, the phosphate analog of fosmidomycin, and its acetyl congener were found to be more potent inhibitors of DXR than fosmidomycin. PMID- 16310361 TI - Synthesis of some novel pyrazolo[3,4-d]pyrimidine derivatives as potential antimicrobial agents. AB - The reaction of 4-hydrazino-8-(trifluoromethyl)quinoline (2) with ethoxymethylenecyanoacetate afforded ethyl 5-amino-1-[8-(trifluoromethyl)quinolin 4-yl]-1H-pyrazole-4-carboxylate (3) and that with ethoxymethylenemalononitrile afforded 5-amino-1-[8-(trifluoromethyl)quinolin-4-yl]-1H-pyrazole-4-carbonitrile (5). Compounds 3 and 5 were hydrolyzed to get 5-amino-1-[8 (trifluoromethyl)quinolin-4-yl]-1H-pyrazole-4-carboxylic acid and then reacted with acetic anhydride to afford 6-methyl-1-[8-(trifluoromethyl)quinolin-4 yl]pyrazolo[3,4-d]oxazin-4-one (6), which was condensed with different aromatic amines to give a series of 5-substituted 6-methyl-1-[8-(trifluoromethyl)quinolin 4-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-ones (7). Compounds 3 and 5 also reacted with formamide, urea, and thiourea affording the corresponding pyrazolo[3,4-d]pyrimidines (8-13), respectively. Structures of the products have been determined by chemical reactions and spectral studies. All compounds of the series have been screened for their antibacterial and antifungal activity studies. The results are summarized in Tables 1 and 2. PMID- 16310362 TI - Temperature stress can alter the photosynthetic efficiency and secondary metabolite concentrations in St. John's wort. AB - Temperature stress is known to cause many physiological, biochemical and molecular changes in plant metabolism and possibly alter the secondary metabolite production in plants. The hypothesis of the current study was that temperature stress can increase the secondary metabolite concentrations in St. John's wort. Plants were grown under controlled environments with artificial light using cool white fluorescent lamps and CO2 enrichment and 70-day-old plants were subjected for 15 days to different temperature treatments of 15, 20, 25, 30 and 35 degrees C before harvested. Major aim of the study was to increase the major secondary metabolites in St. John's wort by applying temperature stress and to evaluate the physiological status of the plant especially the photosynthetic efficiency and peroxidase activity of the leaf tissues exposed to different temperatures under precisely controlled environmental factors. Results revealed that relatively high (35 degrees C) or low (15 degrees C) temperatures reduced the photosynthetic efficiency of the leaves of St. John's wort plants and resulted in low CO2 assimilation. Net photosynthetic rates and the maximal quantum efficiency of PSII photochemistry of the dark adopted leaves (phi(p)max) decreased significantly in the leaves of plants grown under 35 or 15 degrees C temperature treatments. High temperature (35 degrees C) treatment increased the leaf total peroxidase activity and also increased the hypericin, pseudohypericin and hyperforin concentrations in the shoot tissues. These results provide the first indication that temperature is an important environmental factor to optimize the secondary metabolite production in St. John's wort and controlled environment technology can allow the precise application of such specific stresses. PMID- 16310363 TI - Study of cis-cinnamic acid in Arabidopsis thaliana. AB - Trans-cinnamic acid (CA) can be isomerized to cis-CA in Arabidopsis thaliana extract under sunlight. Piperonylic acid treatment of Arabidopsis under ultraviolet (UV) light increased the level of cis-CA in these treated tissues. Similarly, cis-CA was also detected from Oryza sativa seedlings grown under sunlight. These results suggest that cis-CA may occur in planta. Application of cis-CA to seedlings of both wild type Arabidopsis and auxin-insensitive mutants, aux1 and axr2, resulted in nearly identical dose response curves in root growth, indicating that the mode of action by which cis-CA affects plant growth is different from that of auxins. According to root growth inhibition assay, cis-CA is nearly 10 times more active than trans-CA. These results suggest that cis-CA is a unique plant growth regulator but its in vivo function remains to be elucidated. PMID- 16310364 TI - The Arabidopsis inositol 1,3,4-trisphosphate 5/6 kinase, AtItpk-1, is involved in plant photomorphogenesis under red light conditions, possibly via interaction with COP9 signalosome. AB - The COP9 signalosome (CSN) plays important roles in multifaceted cellular processes. Study has shown that inositol 1,3,4-trisphosphate 5/6 kinase (5/6 kinase) interacts with CSN in mammalian cells. However, the biological function of the interaction still remains unknown. Here, we report that the Arabidopsis inositol 1,3,4-trisphosphate 5/6 kinase (AtItpk-1) is also associated with CSN and involved in photomorphogenesis under red light (RL) conditions, as demonstrated by co-immunoprecipitation of AtItpk-1 with CSN and characterization of the atitpk-1 mutants. Expression analysis showed that AtItpk-1 had the same sub-cellular localization and organ expression pattern as CSN. Furthermore, autophosphorylation analysis showed that AtItpk-1 has protein kinase activity. Under RL, the atitpk-1 mutants exhibited phenotype slightly similar with that of the csn mutants, indicating that 5/6 kinase might be involved in the same developmental pathway as CSN. This study suggests that AtItpk-1 may function as a protein kinase that is involved in photomorphogenesis possibly via interaction with COP9 signalosome under red light. PMID- 16310365 TI - Analysis of gun phenotype in barley magnesium chelatase and Mg-protoporphyrin IX monomethyl ester cyclase mutants. AB - The ability of barley (Hordeum vulgare L.) chlorophyll biosynthetic mutants to regulate the expression of Lhc genes was analyzed by a microarray approach. The Lhc genes are located in the nucleus and encode chlorophyll a/b binding proteins of the light-harvesting complex. The chlorophyll a/b binding proteins are some of the many proteins, which are imported to the chloroplast. It has been suggested that the chloroplast can regulate expression of nuclear genes encoding chloroplast proteins, using a chlorophyll biosynthetic intermediate such as Mg protoporphyrin IX (MP) or Mg-protoporphyrin IX monomethyl ester (MPE) as a signal molecule. These compounds are intermediates between the two enzymes magnesium chelatase (EC 6.6.1.1) and Mg-protoporphyrin IX monomethyl ester cyclase (EC 1.14.13.81) in the chlorophyll biosynthetic pathway. Genomes uncoupled (gun) mutants are defective in the chloroplast-to-nucleus signal transduction and express Lhc even when chloroplast development is inhibited by the herbicide norflurazon. We show that barley xantha-f, -g and -h mutants, defective in the three Mg-chelatase genes, have a gun phenotype. In contrast, a xantha-l mutant, defective in a gene of Mg-protoporphyrin monomethyl ester cyclase did not. Genome uncoupling in the xantha-f, -g, -h and -l mutants was also analyzed in absence of norflurazon. All mutants showed transcription of Lhc. This was unexpected in the case of xantha-l as this mutant showed accumulation of MPE, which has been suggested to be one of the two negative regulators of Lhc transcription. We suggest that chlorophyll intermediates may only function as signal molecules at an early developmental stage of chloroplast development. PMID- 16310366 TI - Adaptation of photosynthesis in marama bean Tylosema esculentum (Burchell A. Schreiber) to a high temperature, high radiation, drought-prone environment. AB - Marama bean, Tylosema esculentum, is a tuberous legume native to the Kalahari region of Southern Africa where it grows under high temperatures (typical daily max 37 degrees C during growing season) and radiation (frequently in excess of 2000 micromol m(-2) s(-1)) in sandy soils with low rainfall. These conditions might be expected to select for increased water-use efficiency of photosynthesis. However, marama was found to give similar leaf photosynthetic rates to other C3 plants for a given internal leaf CO2 concentration and Rubisco content. Under conditions of increasing drought, no increase in water-use efficiency of photosynthesis was observed, but stomata closed early and preceded any change in leaf water potential. The possibility of subtle adaptations of photosynthetic characteristics to its natural environment were investigated at the level of Rubisco kinetics. The specificity factor of marama Rubisco was slightly lower than that of wheat, but the apparent Km for CO2 in air (Km') was about 20% lower than that of wheat. This is consistent with better adaptation for efficient photosynthesis at high temperatures in marama compared to wheat, although the net benefit is predicted to be very small (<0.5% at 35 degrees C). The sequence of marama rbcL gene shows 27 deduced amino acid residue differences from that for wheat, and the possibility that one or more of these cause the difference in Rubisco Km' is discussed. PMID- 16310367 TI - Biochemical and molecular characterization of alpha-D-galactosidase from coffee beans. AB - Alpha-D-Galactosidase (alpha-Gal; EC 3.2.1.22) is one of three principal enzymes involved in the modification or degradation of plant cell wall galactomannans. In the present paper it is shown that alpha-galactosidase activities in field-grown coffee beans are variable amongst cultivars of the two species investigated (Coffea arabica and C. canephora var. Robusta). Higher activities were found in Arabica cultivars. Using beans from greenhouse-cultivated C. arabica as a model, we showed that alpha-Gal activity was undetectable in the bean perispem tissue, but increased gradually during the endosperm development, to reach a peak at approximately 30 weeks after flowering (WAF) which coincided with the hardening of the endosperm. Alpha-Gal-specific transcripts detected at 22 and 27 WAF accompanied the peak of alpha-Gal activity, but were reduced to be undetectable in mature beans at 30 WAF, while alpha-Gal activity still persisted. Two isoforms were distinguished in 2-DE profiles of crude protein extracts by N-terminal sequencing analysis. Analysis of two-dimensional gel electrophoresis profiles demonstrated that both isoforms accumulated in a linear fashion throughout grain maturation. Alpha-Gal activity was also observed to increase to high levels during in vitro germination of coffee beans suggesting an important function of this enzyme in this process. Alpha-Gal cDNA sequences from Arabica and Robusta were sequenced and their deduced proteins appeared to be very similar, differing by only eight amino acids. Southern-blot analysis suggests that the enzyme was encoded by at least two genes in C. arabica that could explain the existence of the two isoforms identified in 2-DE profiles. PMID- 16310368 TI - Characterization of lucidin formation in Rubia tinctorum L. AB - In order to approach lucidin formation (a strong mutagen or a carcinogen) from a physiological standpoint, hairy roots of Rubia tinctorum L. were established by a transformation of Agrobacterium rhizogenes strain 15834 and cultured in a liquid woody plant medium without plant hormones. The anthraquinone pigment composition of the intact hairy roots was essentially the same as that of the intact non transformed (normal) roots, in which lucidin O-beta-D-primeveroside (LuP) was one of the major pigments. Lucidin was scarcely detected in the intact hairy roots, but was a main pigment after the squash treatment. The crude protein extract of intact hairy roots exhibited LuP-glycosidase activity (an activity converting LuP to lucidin). This activity was also detected in the roots of the normal plants at a high level, but slightly in the stems and not in the leaves. Methyl jasmonate enhanced the LuP production and LuP-glycosidase activity in the hairy roots. On the other hand, ethephon or salicylic acid had either no effect or rather an inhibitory effect on them. After partial purification of LuP glycosidase, the resultant active fraction producing a major band with an apparent Mr of 68 kDa exhibited the substrate specificity for both aglycon and sugar-moiety. The sugar released from LuP by this fraction was neither D-glucose nor D-xylose and was hydrolyzed into them. These results suggest that LuP specific beta primeverosidase (EC 3.2.1.149) exists in the roots of R. tinctorum and is involved in the systematic defense system. PMID- 16310369 TI - Nod factor induces soybean resistance to powdery mildew. AB - Plants possess highly sensitive perception systems by which microbial signal molecules are recognized. In the Bradyrhizobium-soybean (Glycine max (L.) Merr.) symbiosis, recognition is initiated through exchange of signal molecules, generally flavonoids from soybean and lipo-chitooligosaccharides (Nod factors) from the microsymbiont. Application of the Nod factor Nod Bj-V (C18:1, MeFuc) induced soybean resistance to powdery mildew caused by Microsphaera diffusa. Addition of Nod factor (concentrations ranging from 10(-6) to 10(-10) M) to soybean root systems led to reductions in disease incidence. The lowest disease incidence was caused by Nod factor treatment at 10(-6) M. The effect of Nod factor application on fungal growth and development was measured at 4, 12, 48, and 96 h after inoculation. Colony diameter and number of germ tubes per conidium were decreased by 10(-6) M Nod factor. Phenylalanine ammonia lyase (PAL, EC.4.3.1.1.) is the first enzyme of the phenyl propanoid pathway, and is commonly activated as part of plant responses to disease. Treatment of soybean seedlings with Nod factor, through stem wounds, induced PAL activity; the most rapid increase followed treatment with 10(-6) M Nod factor. These data show that soybean plants are able to detect root applied LCO and respond by increased disease resistance. PMID- 16310370 TI - Effect of intermediates on ascorbic acid and oxalate biosynthesis of rice and in relation to its stress resistance. AB - Rice (Oryza sativa L.) roots were fed with L-ascorbic acid (AsA) and its putative precursors to observe AsA and oxalate concentrations and the resistance of rice to chilling, water stress, and Al toxicity. AsA concentration was significantly enhanced in both shoots and roots of rice seedlings by feeding with D-glucose or L-galactono-gamma-lactone. AsA or L-galactono-gamma-lactone treatment increased accumulation of oxalate mainly in soluble form, while these treatments decreased electrolyte leakage from root cells, H2O2 and lipid peroxidation level in rice seedlings subjected to chilling, water stress, and Al toxicity. They also alleviated the inhibition on root growth by Al. These results indicated that AsA and its immediate precursor protected plants against the oxidative damages induced by various stresses. However, 0.5 mM AsA and 10 mM L-galactono-gamma lactone treatment had no significant effect on superoxide dismutase and catalase activity and ascorbate-peroxidase activities. Enhanced Al resistance caused by AsA and L-galactono-gamma-lactone may possibly be resulted from increased level of oxalate, which acts as metal chelator. Thus it is proposed that manipulation of AsA and oxalate biosynthesis through enhancement of L-galactono-gamma-lactone level in plants could be a strategy for improving abiotic stress tolerance. PMID- 16310371 TI - Cancer of the prostate. AB - Prostate carcinoma, with about 190,000 new cases occurring each year (15% of all cancers in men), is the most frequent cancer among men in northern and western Europe. Causes of the disease are essentially unknown, although hormonal factors are involved, and diet may exert an indirect influence; some genes, potentially involved in hereditary prostate cancer (HPC) have been identified. A suspect of prostate cancer may derive from elevated serum prostate-specific antigen (PSA) values and/or a suspicious digital rectal examination (DRE) finding. For a definitive diagnosis, however, a positive prostate biopsy is requested. Treatment strategy is defined according to initial PSA stage, and grade of the disease and age and general conditions of the patient. In localized disease, watchful waiting is indicated as primary option in patients with well or moderately differentiated tumours and a life expectancy <10 years, while radical prostatectomy and radiotherapy (with or without hormone-therapy) could be appropriate choices in the remaining cases. Hormone-therapy is the treatment of choice, combined with radiotherapy, for locally advanced or bulky disease and is effective, but not curative, in 80-85% of the cases of advanced disease. Patients who develop a hormone-refractory prostate cancer disease (HRPC) have to be evaluated for chemotherapy because of the recent demonstration of improved overall survival (2 2.5 months) and quality of life with docetaxel in more than 1,600 cases. PMID- 16310372 TI - Epidural spinal cord compression. AB - Spinal cord compression from epidural metastases (epidural spinal cord compression, ESCC) is the most common neurological complication of cancer after brain metastases. Extradural compression represents 97% of spinal cord metastatic lesions. ESCC usually occurs in patients with disseminated disease. The most common tumours associated with ESCC are lung and breast cancers, followed by lymphoma, myeloma, prostate cancer and sarcoma. ESCC represents a medical emergency because delayed treatment can be responsible for irreversible deficits, such as paralysis and loss of sphincter control. Patients with ESCC require a multidisciplinary diagnostic and therapeutic approach. Clinical suspect is radiologically detected for confirmation. The median expected survival time from diagnosis usually ranges from 3 to 6 months. The nature of the primary tumour and the degree of the neurological deficit are the most important factors affecting survival. The lack of prospective randomized trials makes the optimal treatment of ESCC controversial and the decision is to be tailored to the individual. Treatment options include: bed rest, administration of corticosteroids, surgery followed by radiation therapy, radiotherapy alone and, to a limited extent, chemotherapy and hormonal therapy. PMID- 16310373 TI - Effect of low molecular lambda-carrageenan from Chondrus ocellatus on antitumor H 22 activity of 5-Fu. AB - Lambda-carrageenan is a kind of sulfated galactan isolated from some red algae and reported to have more biological activities than other types, such as K-type [Guven KC, ozsoy Y, Ulutin ON. Anticoagulant, fibrinolytic and antiaggregant activity of carrageenans and alginic acid. Bot Mar 1991; 34: 429-432]. The molecular weight (MW) of lambda-carrageenan reaches up to 600-700 kDa, the high MW decreased their solubility and limited their bioactivities. At the same time, the production and applications of lambda-carrageenan are limited for its some characteristics, such as the high viscosity, and the experiments of antitumor and immunomodulation also indicated that MW of polysaccharides had notably effect on the activities [Zhou G, Sun YP, Xin H, Zhang Y, Li Z, Xu Z. In vivo antitumor and immunomodulation activities of different molecular weight lambda-carrageenans from Chondrus ocellatus. Pharmacol Res 2004; 50(1) 47-53, Zhou G, Xin H, Sheng W, Sun Y, Li Z, Xu Z. In vivo growth-inhibition of S180 tumor by mixture of 5-Fu and low molecular lambda-carrageenans from C. ocellatus. Pharmacol Res 2005; 51(2) 153-157]. In this research, lambda-carrageenan was extracted from C. ocellatus, an important economic alga in China. Then some small MW samples of 650, 240, 140, 15 and 9.3 kDa are obtained by the method of microwave degradation, respectively. The tumor inhibiting activities of the low MW lambda-carrageenan and mixture of it and 5-Fu on transplanted H-22 tumor mice were investigated. The weight of immune organ, proliferation ratio of lymphocyte and concentration of Tumor Necrosis Factor-alpha (TNF-alpha) and histopathology of tumor from transplanted H 22 tumor mice were also determined. The results indicated that the degraded lambda-carrageenan could add the antitumor activities of 5-Fu and improve the immunocompetence damaged by 5-Fu. PMID- 16310374 TI - Nitric oxide reverses endotoxin-induced inflammatory hyperalgesia via inhibition of prostacyclin production in mice. AB - We examined whether nitric oxide (NO), derived from constitutive NO synthase (NOS) and/or inducible NOS (iNOS), could contribute to endotoxin-induced inflammatory hyperalgesia via interacting with nuclear factor-kappaB (NF-kappaB), inducible cyclooxygenase (COX-2) and/or polyADP-ribose synthase (PARS). Injection of endotoxin (10 mg kg(-1), i.p.) to mice elicited hyperalgesia, determined by hot plate test, which is prevented by NO precursor (L-arginine), cNOS/iNOS inhibitor (N(G)-nitro-L-arginine methyl ester; L-NAME), NF-kappaB inhibitor (N acetylserotonin), COX inhibitor (indomethacin), COX-2 inhibitor (DFU) and PARS inhibitor (3-aminobenzamide). Endotoxin caused a decrease in serum nitrite levels prevented by N-acetylserotonin, L-arginine, indomethacin, DFU or 3 aminobenzamide. Endotoxin increased serum 6-keto-PGF(1alpha) levels diminished by L-arginine or aminoguanidine (iNOS inhibitor). L-Arginine, L-NAME, aminoguanidine, DFU or 3-aminobenzamide prevented endotoxin-induced decrease in heart, lungs, kidneys and brain nitrite and malonedialdehyde levels and myeloperoxidase activity. In conclusion, NO reverses endotoxin-induced inflammatory hyperalgesia via inhibition of prostacyclin production, and also contributes to the analgesic effect of NF-kappaB, COX or PARS inhibitors. PMID- 16310375 TI - An in vitro rat diaphragmatic fatigue model induced by combined hypoxic and hypercapnic acidosis and the effect of salmeterol. AB - Hypoxia or hypercapnia impairs diaphragmatic contractility and induces fatigue. However, little is known about the combined effect of hypoxic and hypercapnic acidosis (HHA) on diaphragmatic fatigue. In this study, a gas mixture (21% O2, 12% CO2 and 67% N2) was used to produce HHA-induced rat diaphragmatic fatigue. Force-frequency relationships and twitch characteristics including peak twitch tension (PTT), time to peak tension (TPT), half relaxation time (1/2RT), maintaining tension (MT) and direct-muscle-stimulation tension (MST) were measured in diaphragm preparations from male SD rats. The HHA gas mixture attenuated force at all frequencies (5-120 Hz) and decreased PTT, MT and MST significantly. Aminophylline, a positive control drug, blocked the negative inotropic effect of HHA in a dose-dependent manner. Moreover, salmeterol, a long acting beta2-adrenoceptor agonist, inhibited the harmful effect of HHA at high frequencies (40-120 Hz), but without effect on MT and MST. These results suggest that an in vitro HHA-induced rat diaphragmatic fatigue model could be established by aerating with the gas mixture, which may be an optimal model to screen effective drugs for diaphragmatic fatigue. Furthermore, salmeterol may play a protective role in HHA-induced impairment. PMID- 16310376 TI - Life before the pre-B cell receptor checkpoint: specification and commitment of primitive lymphoid progenitors in adult bone marrow. AB - The production of B cells is a complex process determined by well-timed combinations of intrinsic factors and environmental cues that guide the differentiation of primitive progenitors in the bone marrow. Expression of several key transcription factors and receptor-stromal cell ligand interactions are landmarks of the earliest events in B lymphopoiesis in adult bone marrow. We describe this as a gradual loss of options for other blood cell lineages coincident with gain of essential properties. Experimental, stress or infection related deregulation may change B cell fate specification, commitment or population dynamics, and consequently the production rate of mature populations. PMID- 16310377 TI - Purification and biochemical characterization of a monomeric form of papaya mosaic potexvirus coat protein. AB - Papaya mosaic virus (PapMV) is a flexuous rod shape virus made of 1400 subunits that assemble around a plus sense genomic RNA. The structure determination of PapMV and of flexuous viruses in general is a major challenge for both NMR and X ray crystallography. In this report, we present the characterization of a truncated version of the PapMV coat protein (CP) that is suitable for NMR study. The deletion of the N-terminal 26 amino acids of the PapMV CP (CP27-215) generates a monomer that can be expressed to high level and easily purified for production of an adequate NMR sample. The RNA gel shift assay showed that CP27 215 lost its ability to bind RNA in vitro, suggesting that the multimerization of the subunit is important for this function. The fusion of a 6x His tag at the C terminus improved the solubility of the monomer and allowed its concentration to 0.2 mM. The CD spectra of the truncated and the wild-type proteins were similar, suggesting that both proteins are well ordered and have a similar secondary structure. CP27-215 was 15N labeled for NMR studies and a 2D 1H-15N-HSQC spectrum confirmed the presence of a well-ordered structure and the monomeric form of the protein. These results show that CP27-215 is amenable to a complete and exhaustive NMR study that should lead to the first three-dimensional structure determination of a flexuous rod shape virus. PMID- 16310378 TI - Improvement in the expression of CYP2B6 by co-expression with molecular chaperones GroES/EL in Escherichia coli. AB - Improvement of CYP2B6 expression was examined by co-expression with molecular chaperones GroES/EL. Although a CO-reduced difference spectrum was not detected in Escherichia coli transformed only by the CYP2B6-expressing vector, co expression of GroES/EL resulted in high-level expression which reached over 2000 nmol P450/L. CYP2B6 was purified from the E. coli membrane with a high yield. Purified CYP2B6 showed 7-ethoxy-4-trifluoromethylcoumarin O-deethylase activity in a reconstitution system. This expression system would be useful for the production of large amounts of active CYP2B6 and for the detailed analysis of the enzyme. PMID- 16310379 TI - Timing of early activity in the visual cortex as revealed by simultaneous MEG and ERG recordings. AB - To clarify the latency of the earliest cortical activity in visual processing, electroretinograms (ERGs) and visual evoked magnetic fields (VEFs) following flash stimulation were recorded simultaneously in six human subjects. Flash stimuli were applied to the right eye and ERGs were recorded from a skin electrode placed on the lower lid. ERGs showed two major deflections in all subjects: an eyelid-negativity around 20 ms and a positivity around 60 ms corresponding to an a- and b-waves, respectively. The mean onset and peak latency of the earliest component of VEFs (37 M) was 30.2 and 36.9 ms, respectively. There was a linear correlation between the peak latency of the a-wave and the onset latency of the 37 M (r=0.90, P=0.011). When a single equivalent current dipole analysis was applied to the 37 M, four out of six subjects showed highly reliable results. The generator of the 37 M was estimated to be located in the striate cortex in all four subjects. Since post-receptoral activities in the retina are expected to start around the peak of the a-wave (20 ms), the early cortical activity, which appears 10 ms later than the a-wave peak, is considered to be the earliest cortical activity following flash stimulation. PMID- 16310380 TI - Mitotic catastrophe and endomitosis in tumour cells: an evolutionary key to a molecular solution. AB - Following genotoxic insult, p53 mutated tumour cells undergo mitotic catastrophe. This is characterised by a switch from mitosis to the endocycle. The essential difference between mitosis and the endocycle is that in the latter, DNA synthesis is uncoupled from cell division, which leads to the formation of endopolyploid cells. Recent data suggests that a return from the endocycle into mitosis is also possible. Furthermore, our observations indicate that a particular type of endocycle known as endomitosis may be involved in this return. Here we review the role of endomitosis in the somatic reduction of polyploidy during development and its postulated role in the evolution of meiosis. Finally, we incorporate these evolutionary data to help interpret our most recent observations in the tumour cell system, which indicate a role for endomitosis and meiotic regulators, in particular p39mos in the segregation of genomes (somatic reduction) of these endopolyploid cells. PMID- 16310381 TI - Identification and characterization of the novel FAD-binding lobe G75S mutation in cytochrome b(5) reductase: an aid to determine recessive congenital methemoglobinemia status in an infant. AB - NADH-cytochrome b(5) reductase deficiency results clinically in either type I or type II recessive congenital methemoglobinemia. The more severe type II form is associated with a global deficiency of cytochrome b(5) reductase and is characterized by cyanosis with neurological dysfunction. In contrast, the only symptom for type I is cyanosis. We have identified a novel G to A mutation at position 15,635 in the DIAI gene of a 4-month-old baby that results in a glycine to serine substitution at codon 75 in the cytochrome b(5) reductase protein. The G75S mutation, located in the FAD-binding lobe of cytochrome b(5) reductase, was found in association with the previously described V252M variant. The V252M mutation is present in the NADH-binding domain and associated with both types I and II recessive congenital methemoglobinemia. Since the G75S and V252M mutations represent radical changes in differing regions of cytochrome b(5) reductase, generating and characterizing these variants singly and in combination using a rat heterologous expression system would provide insight into the differences between types I and II disease at the molecular level. Although all three variants were found to retain stoichiometric levels of FAD with spectroscopic and thermodynamic properties comparable to those of native cytochrome b(5) reductase, all exhibited decreased catalytic efficiency and reduced protein stability reflecting the position of the mutations in the primary structure. The G75S variant retained only 11% of the catalytic efficiency of the wild-type enzyme. Thus, cytochrome b(5) reductase deficient patients who are heterozygous for either FAD- or NADH-binding lobe mutations can exhibit the clinically less severe type I phenotype. PMID- 16310382 TI - A herd level analysis of urinary tract infection in dairy cattle. AB - Cystitis, urethritis and pyelonephritis in cattle most commonly result from ascending urinary tract infection with Corynebacterium renale, Corynebacterium cystidis, Corynebacterium pilosum or Escherichia coli. We describe the clinical, bacteriological, clinical-pathological and epidemiological findings in a dairy cattle herd with urinary tract infection (UTI). Blood and urine samples from 17 calves and 19 cows were submitted to laboratory examinations. Depression, muscle wasting, weakness and frequent urine dribbling were the main characteristics of UTI in calves. Affected cows showed weight loss and an abrupt reduction in feed intake and milk production. Enlargement of the left kidney and loss of normal lobulation were evident on rectal examination. E. coli was the most frequent cause of UTI but C. renale, alpha-haemolytic Streptococcus spp., Proteus spp. Pseudomonas aeruginosa, Klebsiella spp. and Oligella urethralis were isolated as well. Differences in total protein and several protein fractions were found between affected and healthy animals. PMID- 16310383 TI - Evaluation of the virulence of some strains of peste-des-petits-ruminants virus (PPRV) in experimentally infected West African dwarf goats. AB - Different isolates of peste-des-petits-ruminants virus (PPRV) from outbreaks in Africa and India were investigated for virulence in West African dwarf goats in the Ivory Coast. Six groups of five animals received a virulent suspension of various strains of virus at a concentration of 10(3) TCID(50)/mL and the goats were observed for 15 days after infection. The Cote-d'Ivoire 89 (CI89), Guinea Conakry and Bissau Guinea PPRV strains caused a peracute disease; the India Calcutta strain caused acute disease; the Sudan-Sennar strain produced an acute to mild disease, while the Nigeria 75/1 wild-type strain caused a mild disease and the animals recovered. The viruses studied contained examples of PPRV from specific lineage groups based on their nucleoprotein PPRV gene. This experiment indicated that virulence characteristics might be a useful marker to help classify PPRV isolates. PMID- 16310384 TI - A salivary malignant myoepithelioma in a dog. AB - Salivary tumours are uncommon in domestic animals and there are no known previous confirmed reports of salivary tumours of myoepithelial origin in dogs. A 12-year old female mixed breed dog was presented with a lobulated mass, composed of white yellowish tissues, extending from soft palate to epiglottis. Histological examination revealed a neoplastic lesion consisting of a dense population of cells showing moderate pleomorphism, with pale cytoplasm and large oval nuclei, arranged in solid lobules. Mitotic activity was very high. Tumoral cells were negative for both periodic acid-Schiff reaction and Alcian blue stain and displayed strong immunohistochemical reactivity for pan-cytokeratin, muscle specific actin and myosin and focal positivity for cytokeratin 14. On the basis of the morphological, histochemical and immunohistochemical findings a diagnosis of malignant tumour of myoepithelial origin (malignant myoepithelioma) was made. PMID- 16310385 TI - SQUID-detected microtesla MRI in the presence of metal. AB - In magnetic resonance imaging performed at fields of 1 T and above, the presence of a metal insert can distort the image because of susceptibility differences within the sample and modification of the radiofrequency fields by screening currents. Furthermore, it is not feasible to perform nuclear magnetic resonance (NMR) spectroscopy or acquire a magnetic resonance image if the sample is enclosed in a metal container. Both problems can be overcome by substantially lowering the NMR frequency. Using a microtesla imaging system operating at 2.8 kHz, with a superconducting quantum interference device as the signal detector, we have obtained distortion-free images of a phantom containing a titanium bar and three-dimensional images of an object enclosed in an aluminum can; in both cases high-field images are inaccessible. PMID- 16310386 TI - A putative bioactive conformation for the altered peptide ligand of myelin basic protein and inhibitor of experimental autoimmune encephalomyelitis [Arg91, Ala96] MBP87-99. AB - [Arg(91), Ala(96)] MBP(87-99) is an altered peptide ligand (APL) of myelin basic protein (MBP), shown to actively inhibit experimental autoimmune encephalomyelitis (EAE), which is studied as a model of multiple sclerosis (MS). The APL has been rationally designed by substituting two of the critical residues for recognition by the T-cell receptor. A conformational analysis of the APL has been sought using a combination of 2D NOESY nuclear magnetic resonance (NMR) experiments and detailed molecular dynamics (MD) calculations, in order to comprehend the stereoelectronic requirements for antagonistic activity, and to propose a putative bioactive conformation based on spatial proximities of the native peptide in the crystal structure. The proposed structure presents backbone similarity with the native peptide especially at the N-terminus, which is important for major histocompatibility complex (MHC) binding. Primary (Val(87), Phe(90)) and secondary (Asn(92), Ile(93), Thr(95)) MHC anchors occupy the same region in space, whereas T-cell receptor (TCR) contacts (His(88), Phe(89)) have different orientation between the two structures. A possible explanation, thus, of the antagonistic activity of the APL is that it binds to MHC, preventing the binding of myelin epitopes, but it fails to activate the TCR and hence to trigger the immunologic response. NMR experiments coupled with theoretical calculations are found to be in agreement with X-ray crystallography data and open an avenue for the design and synthesis of novel peptide restricted analogues as well as peptide mimetics that rises as an ultimate goal. PMID- 16310387 TI - Combined electronic-topological and neural networks study of some hydroxysemicarbazides as potential antitumor agents. AB - Structure-activity relationships study was performed for a series of Schiff bases hydroxysemicarbazide as potential antitumor agents by using the electronic topological method combined with neural networks (ETM-NN). Data for the approach were obtained from conformational and quantum-chemical calculations and arranged first as matrices called electronic-topological matrices of contiguity, by one for each compound. Then specific molecular fragments were found for active compounds ('activity features') from the ETM application. After this, a system of prognosis was developed as the result of training the Kohonen self-organizing maps (SOM) by the most significant fragments. PMID- 16310388 TI - Comparable efficacy of ciclesonide once daily versus fluticasone propionate twice daily in asthma. AB - BACKGROUND: Inhaled corticosteroids are the mainstay of therapy in asthma, but local and systemic side effects and adherence remain a concern. Ciclesonide is an inhaled corticosteroid with on-site lung activation that provides potent anti inflammatory activity and has been shown to have a good safety profile, even at high doses. OBJECTIVE: The aim of this study was to compare the efficacy and safety of once-daily ciclesonide versus twice-daily fluticasone propionate at comparable daily doses in patients with asthma. METHODS: In this multicenter, randomized, double-blind, double-dummy, parallel group study, 529 patients were randomized to ciclesonide 160 microg once daily or fluticasone propionate 88 microg twice daily for 12 weeks. The primary endpoint was change in lung function. RESULTS: Both ciclesonide and fluticasone propionate significantly improved forced expiratory volume in 1s, forced vital capacity, and morning peak expiratory flow compared with baseline (p<0.0001 for all variables). Both medications reduced asthma symptoms and rescue medication use within the first 24 h. At the tested dose, both medications were equally safe and well tolerated. CONCLUSION: Ciclesonide 160 microg once daily was as effective as fluticasone propionate 88 microg twice daily in improving lung function and asthma symptoms, and in reducing rescue medication use in patients with asthma. PMID- 16310389 TI - Highlights on the appropriate use of fluoroquinolones in respiratory tract infections. AB - The impact of respiratory infections on public health is increasing, and lower respiratory tract infections are a major cause of morbidity and mortality. Moreover, most antibiotic prescriptions are related to respiratory infections and this is probably one of the main determinants of the increasing rate of bacterial resistance in both community and hospital settings. This has been the catalyst for the development of new drugs, such as the new fluoroquinolones. The new fluoroquinolones have an excellent spectrum providing cover for the most important respiratory pathogens, including atypical and "typical" pathogens. The pharmacokinetic and dynamic properties of the new fluoroquinolones have a significant impact on their clinical and bacteriological efficacy. They cause a concentration-dependent killing with a sustained post-antibiotic effect. Fluoroquinolones combine exceptional efficacy with cost-effectiveness. Not surprisingly, different guidelines have inserted these agents among the drugs of choice in the empirical therapy of LRTIs. This review discusses the most recent data on the bacteriological and clinical activity of the new fluoroquinolones and critically analyses the risks of a potential overuse of this valuable new class of drugs. PMID- 16310390 TI - The isolated anterior stomach of larval mosquitoes (Aedes aegypti): voltage-clamp measurements with a tubular epithelium. AB - The anterior stomach of larval Aedes aegypti was isolated and perfused via two pipettes. For transepithelial voltage (V(te)) measurement, the inflow pipette and the bath were connected via agar bridges to calomel electrodes. For voltage clamping, the lumen of the tissue contained an Ag/AgCl wire held by the outflow pipette, and the preparation was placed in a bath within a spiral of Ag/AgCl wire. After equilibrating the tissue in mosquito saline on both sides, a V(te) of -8+/-1 mV was measured (+/-S.E.M., N=32). Current-voltage curves (+/-100 mV) demonstrated ohmic behaviour of the epithelium. Short-circuiting resulted in a current (I(sc)) of 103+/-16 microA cm(-2) and a mean transepithelial conductance (G(te)) of 11.8+/-1.3 mS cm(-2) (+/-S.E.M., N=32). A Yonath-Civan plot of G(te) of individual preparations over the corresponding I(sc) resulted in a straight line (r(2)=0.8422), indicating that the difference in I(sc) of individual preparations is mainly based on different transcellular conductances (G(c)). This analysis allowed to estimate the mean leak conductance (G(l) approximately 3.9 mS cm(-2)) and the mean transcellular electromotive force (E(c) approximately 13 mV). After administering 0.2 micromol L(-1) serotonin, I(sc) and G(te) significantly increased, to 457+/-49 microA cm(-2) and to 21.3+/-2.3 mS cm(-2) (+/-S.E.M., N=31, P<0.05), respectively. The Yonath-Civan plot after serotonin resulted again in a straight line (r(2)=0.8219), indicating a mean G(l) of about 1 mS cm(-2) and a mean E(c) of about 22 mV. Dinitrophenol (2.5 mmol L(-1)) almost abolished I(sc) and significantly reduced G(te) (N=6). Concanamycin A (100 micromol L(-1)) reduced I(sc) by more than 90% without significantly affecting G(te). PMID- 16310391 TI - Analysis of 5-methyltetrahydrofolate in serum of healthy children. AB - 5-Methyltetrahydrofolate (5MTHF) is the active one-carbon donor and the principal circulating form of plasma folates. It is involved in a number of metabolic and neurodevelopmental processes and analysis of cerebrospinal fluid (CSF) 5MTHF is of great importance in the diagnosis of cerebral folate deficiency (CFD). Serum 5MTHF levels and the 5MTHF serum/CSF ratio may be important additional parameters for the understanding of CFD. We developed a HPLC method for the measurement of 5MTHF in serum and established reference values for the pediatric population. Serum samples from 64 healthy children were extracted with Sep-Pak C18 cartridges and 5MTHF was separated by RP-HPLC and quantified by electrochemical detection. 5MTHF was separated from other folates and detected after 8.7 min with linearity of up to 1600 nmol/L. The detection limit was 4.5 nmol/L and recovery during solid-phase extraction for low and high concentrations of 5MTHF was 66 and 62%, respectively. Within-run imprecision (13.5%) was slightly higher than run-to-run imprecision (8.5%). 5MTHF levels in healthy children were found to be age dependent, decreasing from 158.0 nmol/L in newborns to 60.1 nmol/L in children older than 16 years. The method we describe is sensitive, selective, and reliable for the analysis of 5MTHF from 400 microL of serum. PMID- 16310392 TI - gamma-H2AX dephosphorylation by protein phosphatase 2A facilitates DNA double strand break repair. AB - Phosphorylated histone H2AX (gamma-H2AX) forms foci over large chromatin domains surrounding double-stranded DNA breaks (DSB). These foci recruit DSB repair proteins and dissolve during or after repair is completed. How gamma-H2AX is removed from chromatin remains unknown. Here, we show that protein phosphatase 2A (PP2A) is involved in removing gamma-H2AX foci. The PP2A catalytic subunit [PP2A(C)] and gamma-H2AX coimmunoprecipitate and colocalize in DNA damage foci and PP2A dephosphorylates gamma-H2AX in vitro. The recruitment of PP2A(C) to DNA damage foci is H2AX dependent. When PP2A(C) is inhibited or silenced by RNA interference, gamma-H2AX foci persist, DNA repair is inefficient, and cells are hypersensitive to DNA damage. The effect of PP2A on gamma-H2AX levels is independent of ATM, ATR, or DNA-PK activity. PMID- 16310393 TI - Brain abscess: clinical aspects of 100 patients. AB - OBJECTIVE: To verify if, in the last two decades, there have been any changes in epidemiological, clinical, diagnostic, therapeutic and prognostic aspects of patients with brain abscess. METHOD: We studied retrospectively 100 patients discharged over a 17-year period from Ca' Foncello Regional Hospital, Treviso, Italy with a diagnosis of brain abscess. RESULTS: Post-surgical abscesses were more frequent than those related to contiguous infections and the spectrum of etiologic agents was very heterogeneous. A cerebral neoplasm was the initial neuroradiological diagnosis in 13 patients; 72 patients underwent a neurosurgical procedure. A comatose state at presentation was associated with an unfavourable outcome. CONCLUSION: With the exception of some epidemiological aspects, which varied from the literature, in spite of the improvements in diagnostic procedures and treatment, no significant changes occurred in the prognosis of patients with brain abscess. PMID- 16310394 TI - Spontaneous, nontraumatic gas gangrene due to Clostridium perfringens. PMID- 16310395 TI - Determination of penicillin resistance in Streptococcus pneumoniae isolates from southern Brazil by PCR. AB - OBJECTIVE: To demonstrate the potential clinical applicability of the PCR technique to the early detection of bacterial resistance in Streptococcus pneumoniae. METHODS: We studied 153 samples of S. pneumoniae, isolated from different anatomic sites, using polymerase chain reaction (PCR) for the detection of specific amplicons from genes that code for penicillin-binding proteins (PBP) 1a, 2b and 2x, which are responsible for penicillin resistance in this organism. The occurrence of these mutated genes was correlated with the minimum inhibitory concentration (MIC) of penicillin, determined by the agar dilution test. RESULTS: The rate of penicillin resistance in S. pneumoniae in Porto Alegre, Brazil was 22.8% (16.3% intermediate resistance and 6.5% high resistance). In a statistically significant proportion of cases (p < 0.05), penicillin-susceptible samples had no amplicons, intermediate samples had only one (generally from PBP 2x), and highly resistant samples had amplicons from all three PBPs investigated. CONCLUSION: These results suggest that penicillin resistance in S. pneumoniae in southern Brazil is on the increase, but is still lower than in other countries, and that PCR could be used for its early detection. PMID- 16310396 TI - [Soft tissue sarcomas: current data in the field of pathology]. AB - Over the last fifteen years, pathology underwent significant changes in the field of soft tissue tumours. They were related to considerable advances in molecular biology and genetics. New data led to the revision of the WHO classification. Malignant fibrous histiocytoma is no longer considered as an entity. It has split up into several subgroups belonging to liposarcomas, leiomyosarcomas or undifferentiated sarcomas. Haemangiopericytoma underwent reappraisal and was put in the same category as solitary fibrous tumour. Many tools have improved. Immunohistochemistry performed with new antibodies had its specificity increased, and became appropriate for the prediction of therapeutic response in some cases, e.g. CD117 detecting mutations of the c-kit proto-oncogen in gastro-intestinal stromal tumours. Refinement of the techniques allows accurate diagnoses from core needle biopsies. Surgical specimens are collegially examined by surgeons and pathologists with special attention paid to resection margins. Although bound by some limitations, the grading system of the French Federation of Cancer Centers has currently remained the best predictor of metastasis-free survival and overall survival of patients. It is based on an assessment of three parameters: differentiation, amount of necrosis, and mitotic count of tumours. The pathologist sets up a diagnosis, and actively takes part in the prediction of the prognosis and therapeutic response. He is one of the major participants in decision making for multimodal treatment of sarcomas. PMID- 16310397 TI - [Assessment of tumor radiosensitivity using functional and metabolic nuclear imaging in research and clinical practice. A review]. AB - During the last half of century considerable research on radiosensitivity biomarkers has been published. However, to date there is no non-invasive marker of cellular radiosensitivity identified for clinical routinely use. In this review, the main functional and metabolic imaging isotopic techniques for tumor radiosensitivity that have been explored over the last years are being described. This indirect evaluation fall into 3 topics associated with tumor proliferation rate or apoptosis, tumor hypoxic fraction, neoangiogenesis and the intrinsic radiosensitivity of clonogenic tumor cells. The final objective of the radiosensitivity monitoring during radiotherapy would be to adapt treatment strategy for overcoming the identified radioresistance mechanism such as hypoxia by the addition of radiosensitisers for example. This would allow better tumor control rather than continue inefficient and costly treatment delivery, which in addition could compromise outcome. PMID- 16310398 TI - Psoriatic arthritis. AB - Psoriatic arthritis exhibits specific characteristics that create challenges for physicians attempting to manage the disease. Active research has produced new knowledge in the fields of epidemiology, diagnostic criteria, evaluation tools, outcome prediction, and treatment. Biotherapies have been introduced for psoriatic arthritis, creating a need for a reappraisal of evaluation tools, outcome prediction, and parameters reflecting treatment effects. TNF-alpha antagonists constitute a breakthrough in the management of psoriatic arthritis. Similar to rheumatoid arthritis and ankylosing spondylitis, psoriatic arthritis is entering a new era of evaluation and treatment. PMID- 16310399 TI - Forensic considerations of surrogacy -- an overview. AB - In the past two decades, there has been a spectacular change in the field of reproductive technologies. Reproductive sciences have come in with techniques such as donor insemination; in vitro fertilization and embryo transfer methods, which have completely revolutionized the reproductive environment. These techniques have given hope to many infertile couples, who wish to have a child of their own. With the oncoming of surrogacy agreements, concepts of fatherhood and motherhood have been subject to much controversy. Motherhood was never under much scrutiny as it was rightly thought that childbirth was the natural and conclusive fact establishing it, though paternity was at times a controversial concept burdening legislators to provide for suitable tests. Medical sciences have now compartmentalized motherhood into the genetic, gestational and the social mother leading to a clash of interests. This paper reviews some medicolegal, ethical and social aspects of surrogacy. PMID- 16310400 TI - Paternal and sibling incest: a case report. AB - A case is reported of a female victim of paternal incest, who had also been raped repeatedly by her elder brother for two years. A survey of the literature showed no other report of such a case from Turkey. This does not necessarily mean that the incidence of paternal and sibling incest does not happen, but may indicate that incestuous abuse is not reported or handled without making it known to legal authorities. The victim was first raped by her 16 year-old brother when she was 9 years old. He raped her repeatedly over a period of two years, until he left home. Her father began raping the victim when she was 13 year-old, leaving her pregnant at age 15. He took her to a doctor for a termination of pregnancy. The father continued abuse after the termination. The victim left home to marry a man. The father filed a lawsuit against the man for taking the victim away from home. More openness and awareness of incest in Turkey may encourage the victims to seek help from medical and legal authorities. PMID- 16310401 TI - Potent histamine-releasing activity of atrahagin, a novel snake venom metalloproteinase. AB - Poisonous snakebite wound is a popular disease worldwide. However, the pathogenesis remains unclear. In the present study, a novel metalloproteinase atrahagin in Chinese cobra (Naja atra) snake venom was purified, using heparin sepharose followed by Superdex 75 gel filtration chromatography. Apart from its alpha-fibrinogenase activity, atrahagin potently activated human colon, lung and tonsil mast cells with the net histamine release being 25.9+/-4.4, 17.0+/-1.9, 13.2+/-3.6%, respectively. Time course studies revealed that the peak histamine release induced by atrahagin occurred at 12, 12 and 8 min following incubation of the enzyme with colon, lung and tonsil mast cells, respectively. The response of mast cells to atrahagin was abolished by preincubation of the cells with metabolic inhibitors or pertussis toxin, and by removal of Ca2+ and Mg2+ from the challenge buffer. In conclusion, activation of human mast cells by atrahagin indicated that the enzyme might contribute to the pathogenesis of snakebite wound. PMID- 16310402 TI - Alterations in components of the TGF-beta superfamily signaling pathways in human cancer. AB - Signaling by transforming growth factor-beta (TGF-beta) superfamily ligands to the nucleus is mediated by type I and type II receptors and the intracellular signal transducers, the Smads. Alteration of some of the components of these pathways has been observed in human tumors. These alterations can be deletions or mutations, or downregulation of components that act positively in the pathway, or alternatively, amplification or overexpression of inhibitors of the pathways. The selection of these alterations during tumor progression and their correlation with clinical outcomes, such as survival, risk of recurrence after tumor resection or tendency for metastatic spread, suggest that many are involved in tumor progression. Here, we review the genetic alterations and epigenetic modifications that occur in different components of the TGF-beta superfamily signaling pathways in human tumors and we discuss their correlation with clinical outcome. The evidence suggests that not all alterations of the TGF-beta superfamily signaling pathway components in human cancer have an equivalent effect on tumor progression and we discuss what implications this has for our understanding of the role of TGF-beta signaling in human cancer. PMID- 16310403 TI - Comparison of the relative sensitivity of human lymphocytes and mouse splenocytes to two spindle poisons. AB - Aneugenic compounds act on non-DNA targets to exert genotoxicity via an indirect mechanism. In contrast to DNA-binding agents, these compounds are expected to possess threshold levels of activity. Therefore, the risk for adverse effects following human exposure to an aneugen could be minimal, if the threshold of activity has been clearly determined in vivo and in vitro and providing the human exposure level is below this threshold. Thus, the development of a single-cell model to allow comparisons between in vitro and in vivo threshold values for aneugenic compounds is of importance. The in vivo micronucleus test is one of the main assays used in genetic toxicology, and is often performed in the mouse. Thus, an extensive database is available in the literature. However, there are only few data concerning the in vitro micronucleus assay using mouse cells, as the majority of in vitro micronucleus assays have been performed using human lymphocytes. In addition, there is a lack of data concerning thresholds for any compound using this model. First, we evaluated whether the use of mouse splenocytes would be an acceptable alternative to that of human lymphocytes to identify aneugens. To allow valid comparisons, the two protocols were first harmonized. Thus, phytohemagglutinin (PHA) and concanavalin A were used as specific mitogens for human lymphocytes and mouse splenocytes, respectively, in order to achieve similar cell-proliferation rates. To achieve similar and sufficient numbers of binucleated cells, cytochalasin B was added 44 and 56 h after culture initiation of the human and mouse cells, respectively. Second, we compared the sensitivity of the mouse protocol with that of the human protocol by exposing the cells to the aneugens nocodazole and paclitaxel. There was good reproducibility of the cytotoxic/genotoxic responses of the two cell models following exposure to the aneugens. The sensitivity of the mouse splenocytes to paclitaxel was higher than that of the human lymphocytes. The two cell types were equally sensitive to nocodazole. PMID- 16310404 TI - A simple and fast densitometric method for the analysis of tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta and in the ventral tegmental area. AB - Parkinson's disease is a progressive dyskinetic disorder caused by degeneration of mesencephalic dopaminergic neurons in the substantia nigra pars compacta (SNpc) and, to a lesser extent, in the ventral tegmental area (VTA). Tyrosine hydroxylase (TH) is a rate-limiting enzyme for dopamine synthesis, therefore immunohistochemistry for TH can be used as an important marker of dopaminergic cell loss in these regions. Traditionally, immunohistochemical experiments are analyzed qualitatively by optical microscopic observation or more rarely semi quantitatively evaluated by densitometry. A common problem with such papers is the lack of a clear explanation of the algorithms and macros employed in the semi quantitative approaches. In this paper, we describe, in detail, an easy, fast and precise protocol for the analysis of TH immunoreactivity in SNpc and VTA using one of the most popular image analysis software packages (Image Pro-Plus). We believe that this protocol will facilitate the evaluation of mesencephalic TH immunoreactivity in various available animal models of Parkinson's disease. PMID- 16310405 TI - Rubella in pregnancy: intrauterine transmission and perinatal outcome during a Brazilian epidemic. AB - Sixty pregnant women with clinical signs of rubella and specific rubella antibodies were studied between January 1999 and December 2002 in order to determine the intrauterine rubella transmission rate and the presence of the virus in amniotic fluid and fetal tissues by nested PCR. Thirty-three patients presented rubella before 12 weeks of gestation and 27 after 12 weeks. Gestational age at the time of acute rubella was determined on the basis of the date of last menstruation and the first trimester ultrasound scan. Thirteen patients with clinical features of rubella before 12 weeks of gestation were submitted to amniocentesis. Three products of conception were analyzed. The presence of the rubella virus was determined by nested PCR. IgM and IgG antibodies were analyzed in neonatal samples at birth and at 3 months of age using a capture immunoassay. Newborn follow-up was based on the presence of congenital rubella syndrome compatible defects, anti-rubella antibodies, echocardiographic alterations, brainstem evoked response audiometry, and ophthalmological pathology. Five miscarriages and four fetal deaths were observed in the group of patients presenting clinical features before 12 weeks of gestation. IgM antibodies were detected in seven neonates at birth and at 3 months of age. Deafness was observed in three cases and pigmentary retinopathy in one case. Fourteen of the 16 samples (13 amniotic fluid and 3 fetal tissue samples) submitted to virological analysis tested positive. Four fetal deaths, five miscarriages (one with negative virology) and seven newborns with anti-rubella IgM at birth and/or at 3 months age were observed in the group with rubella before 12 weeks of gestation. There were three cases in which virological analysis of the amniotic fluid samples was positive (infected) while the newborn showed no signs of congenital rubella syndrome and anti-rubella IgM were absent. When maternal rubella occurred after 12 weeks of gestation, no fetal or neonatal rubella signs were observed. Eradication of congenital rubella syndrome is possible since vaccination campaigns continue and all services related to the health care of children, adolescents and women have become aware of the significance of the problem and are collaborating. All pregnant women in Brazil should be screened for the rubella antibody and the susceptible group should be vaccinated after giving birth. PMID- 16310406 TI - Simultaneous detection and typing of human metapneumovirus strains in nasopharyngeal secretions and cell cultures by monoclonal antibodies. PMID- 16310407 TI - Vasoactive intestinal peptide--release from the heart and response in heart failure due to left ventricular pressure overload. AB - BACKGROUND: Vasoactive intestinal peptide (VIP) is a peptidergic neurotransmitter and a vasodilator with positive inotropic and chronotropic properties. Whether and how VIP contributes to the neuroendocrine response in heart failure (HF) is disputed, and there are no data on VIP in pressure overload-induced HF. METHODS: We studied 129 adults with isolated aortic valve stenosis (AS). Blood was sampled from the aortic root and, in a subset of 48 patients, also from the coronary sinus for determination of VIP by radioimmunoassay. HF was diagnosed according to the European Society of Cardiology criteria. RESULTS: Plasma VIP (mean+/-S.E.M.) was slightly higher in patients with HF (22.6+/-0.9 pmol/l, n=41) than in patients free of HF (21.1+/-0.5 pmol/l, n=88) or in 11 control patients without structural heart disease (20.0+/-1.3 pmol/l, n=11) (p=0.030 across the groups). VIP did not correlate with any measurement of cardiac structure or function in AS. The change in plasma VIP from aortic root to coronary sinus averaged +1.2+/ 0.4 pmol/l in the 11 control patients (p=0.021), +1.2+/-0.2 pmol/l in 33 AS patients free of HF (p<0.001) and +0.8+/-0.3 pmol/l in 15 AS patients with HF (p=0.037). CONCLUSIONS: Both structurally normal and diseased hearts release VIP into the coronary sinus. Although marginally elevated in the systemic circulation, VIP is unlikely to contribute significantly to the neuroendocrine activation in HF due to pressure overload. PMID- 16310408 TI - VE/VCO2 slope is associated with abnormal resting haemodynamics and is a predictor of long-term survival in chronic heart failure. AB - BACKGROUND: Patients with chronic heart failure (CHF) present with exercise induced hyperpnea, but its pathophysiological mechanism has not been thoroughly investigated. We aimed to determine the relationship between exercise-induced hyperpnea, resting haemodynamic measurements and the validity of ventilatory response (V(E)/V(CO(2)) slope) as a mortality predictor in CHF patients. METHODS: Ninety-eight CHF patients (90M/8F) underwent a symptom-limited treadmill cardiopulmonary exercise test (CPET). Right heart catheterization and radionuclide ventriculography were performed within 72 h of CPET. RESULTS: Twenty seven patients died from cardiac causes during 20+/-6 months follow-up. Non survivors had a lower peak oxygen consumption (V(O(2)p)), (16.5+/-4.9 vs. 20.2+/ 6.1, ml/kg/min, p=0.003), a steeper V(E)/V(CO(2)) slope (34.8+/-8.3 vs. 28.9+/ 4.8, p<0.001) and a higher pulmonary capillary wedge pressure (PCWP) (19.5+/-8.6 vs. 11.7+/-6.5 mm Hg, p=0.008) than survivors. By multivariate survival analysis, the V(E)/V(CO(2)) slope as a continuous variable was an independent prognostic factor (chi(2): 8.5, relative risk: 1.1, 95% CI: 1.03-1.18, p=0.004). Overall mortality was 52% in patients with V(E)/V(CO(2)) slope > or =34 and 18% in those with V(E)/V(CO(2)) slope <34 (log rank: 18.5, p<0.001). In a subgroup of patients (V(O(2)p): 10-18 ml/kg/min), V(E)/V(CO(2)) slope was a significant predictor of mortality (relative risk: 6.2, 95% CI: 1.7-22.2, p=0.002). Patients with high V(E)/V(CO(2)) slope had higher resting PCWP (19.9+/-9.1 vs. 11.3+/-5.7 mmHg, p<0.001) and V(E)/V(CO(2)) slope correlated significantly with PCWP (r: 0.57, p<0.001). CONCLUSIONS: The V(E)/V(CO(2)) slope, as an index of ventilatory response to exercise, improves the risk stratification of CHF patients. Interstitial pulmonary oedema may be a pathophysiological mechanism of inefficient ventilation during exercise in these patients. PMID- 16310409 TI - Obstructive sleep apnea: re-evaluating our index of severity. PMID- 16310410 TI - The neurophysiology of the alternating leg muscle activation (ALMA) during sleep: study of one patient before and after treatment with pramipexole. AB - BACKGROUND AND PURPOSES: To report the neurophysiological features of a patient with alternating leg muscle activation (ALMA) during sleep, a quickly alternating pattern of anterior tibialis activation which might represent transient facilitation of a spinal central pattern generator for locomotion, perhaps due to the serotonergic effects of antidepressant medication. PATIENT AND METHODS: A 33 year-old male patient with ALMA. The patient underwent a complete and detailed study of his neurophysiological parameters during sleep, before and after treatment with pramipexole. RESULTS: The treatment with pramipexole was followed by a significant reduction in the rate of occurrence of ALMA, in reported insomnia, and in daytime sleepiness. The ALMA generally were preceded by cyclic alternating pattern A phases and increased heart rate in most instances. Visual scoring and spectral analyses suggested that after pramipexole more intense arousal was required to trigger ALMA. CONCLUSION: The evident beneficial effect induced by the treatment with pramipexole indicates that the spinal networks involved in the generation of ALMA might also be under the inhibitory control of dopaminergic networks. We suggest that ALMA can be seen even in the absence of other factors such as antidepressant therapy, sleep apnea or periodic leg movements during sleep, and might be considered as an additional phenomenon influenced by sleep instability. Our patient seems to indicate also that treatment with dopamine agonists can be useful in such patients because the treatment can be followed by a good clinical response. PMID- 16310411 TI - Triggering TLR signaling in vaccination. AB - Toll-like receptors (TLRs) are a family of pattern-recognition receptors that are an important link between innate and adaptive immunity. Many established, as well as experimental, vaccines incorporate ligands for TLRs, not only to protect against infectious diseases but also in therapeutic immunization against noninfectious diseases, such as cancer. We review the underlying mechanisms by which engagement of TLR signaling pathways might trigger an adaptive immune response after immunization. Although the engagement of TLR signaling pathways is a promising mechanism for boosting vaccine responses, questions of efficacy, feasibility and safety remain the subject of active investigation. PMID- 16310412 TI - Coevolutionary interactions between host and parasite genotypes. AB - More than 20 years after Dawkins introduced the concept of "extended phenotype" (i.e. phenotypes of hosts and parasites result from interactions between the two genomes) and although this idea has now reached contemporary textbooks of evolutionary biology, most studies of the evolution of host-parasite systems still focus solely on either the host or the parasite, neglecting the role of the other partner. It is important to consider that host and parasite genotypes share control of the epidemiological parameters of their relationship. Moreover, not only the traits of the infection but also the genetic correlations among these and other traits that determine fitness might be controlled by interactions between host and parasite genotypes. PMID- 16310413 TI - Chemokines in host-parasite interactions in leishmaniasis. AB - Crucial to the defense against leishmaniasis is the ability of the host to mount a cell-mediated immune response capable of controlling and/or eliminating the parasite. Cell recruitment to the site of infection is essential to the development of the host cellular immune response. The process is controlled by chemokines, which are chemotactic cytokines produced by leukocytes and tissue cells. PMID- 16310414 TI - Aging of mesenchymal stem cells. AB - The role of adult mesenchymal stem cells (MSC) in tissue maintenance and regeneration has received significant attention of late. Questions arise to what extent these cells are either subject to, or causes of aging; whether age-related changes in these cells are due to intrinsic factors or induced by the somatic environment. This review collates and examines recent data in support of these different theories. By means of introduction, a brief overview is given of current MSC definitions and their basic role in tissue regeneration followed by a comparative analysis of gerontological studies involving MSC. Evidence for extrinsic aging and various aging markers relating to morphology, proliferation, signalling, senescence markers, telomeres and telomerase, and other indicators is discussed. We observe that while the literature might often appear to conflict, many apparent discrepancies are attributable to inconsistent methods of extracting and isolating MSC which in fact contains various subsets of adult stem cells, varying not only in their differentiation potential but also in their vulnerability to senescence--ranging from quasi-somatic lifespan to perennial vigour. Thus, mesenchymal stem cells emerge as both subject to and key mediators of organismal aging. PMID- 16310415 TI - Roles of E. coli double-strand-break-repair proteins in stress-induced mutation. AB - Special mechanisms of mutation are induced during growth-limiting stress and can generate adaptive mutations that permit growth. These mechanisms may provide improved models for mutagenesis in antibiotic resistance, evolution of pathogens, cancer progression and chemotherapy resistance. Stress-induced reversion of an Escherichia coli episomal lac frameshift allele specifically requires DNA double strand-break-repair (DSBR) proteins, the SOS DNA-damage response and its error prone DNA polymerase, DinB. We distinguished two possible roles for the DSBR proteins. Each might act solely upstream of SOS, to create single-strand DNA that induces SOS. This could upregulate DinB and enhance mutation globally. Or any or all of them might function other than or in addition to SOS promotion, for example, directly in error-prone DSBR. We report that in cells with SOS genes derepressed constitutively, RecA, RuvA, RuvB, RuvC, RecF, and TraI remain required for stress-induced mutation, demonstrating that these proteins act other than via SOS induction. RecA and TraI also act by promoting SOS. These and additional results with hyper-mutating recD and recG mutants support roles for these proteins via error-prone DSBR. Such mechanisms could localize stress induced mutagenesis to small genomic regions, a potentially important strategy for adaptive evolution, both for reducing additional deleterious mutations in rare adaptive mutants and for concerted evolution of genes. PMID- 16310416 TI - Serotoninergic receptors in the anteroventral preoptic region modulate the hypoxic ventilatory response. AB - Hypothalamus is a site of integration of the hypoxic and thermal stimuli on breathing and there is evidence that serotonin (5-HT) receptors in the anteroventral preoptic region (AVPO) mediate hypoxic hypothermia. Once 5-HT is involved in the hypoxic ventilatory response (HVR), we investigated the participation of the 5-HT receptors (5-HT1, 5-HT2 and 5-HT7) in the AVPO in the HVR. To this end, pulmonary ventilation (V(E)) of rats was measured before and after intra-AVPO microinjection of methysergide (a 5-HT1 and 5-HT2 receptor antagonist), WAY-100635 (a 5-HT1A receptor antagonist) and SB-269970 (a 5-HT7 receptor antagonist), followed by 60 min of hypoxia exposure (7% O2). Intra-AVPO microinjection of vehicles or 5-HT antagonists did not change V(E) during normoxic conditions. Exposure of rats to 7% O2 evoked typical hypoxia-induced hyperpnea after vehicle microinjection, which was not affected by methysergide. WAY-100635 and SB-269970 treatment caused an increased HVR, due to a higher tidal volume. Therefore, the current data provide the evidence that 5-HT acting on 5 HT1A and 5-HT7 receptors in the AVPO exert an inhibitory modulation on the HVR. PMID- 16310417 TI - A new on-line, in-tube pre-column derivatization technique for high performance liquid chromatographic determination of azithromycin in human serum. AB - Pre-column derivatization methods for high performance liquid chromatographic assay of specific pharmaceutical agents using 9-fluorenylmethyl chloroformate (FMOC-Cl) have received special attention because highly fluorescent and stable adducts are provided by these methods. However, unlike the post-column on-line techniques, long derivatization time is needed and the reaction cannot be well controlled. A new, sensitive and fast pre-column on-line derivatization technique coupled with high-performance liquid chromatography using FMOC-Cl as labeling agent is described and validated for determination of azithromycin in human serum. After extraction of the drug from serum, the residue was reconstituted in mixture of acetonitrile-phosphate buffer (3:1, v/v; pH 8.5) and directly injected onto the chromatographic system. Continuous on-line derivatization and analysis of the compounds were successfully performed using in-tube elution of FMOC-Cl. The total time needed for derivatization and chromatographic analysis of the drug was 13 min. The assay was reliable and reproducible, with limit of quantification of 10 ng/ml. The described technique may offer significant advantages over existing off-line derivatization methods using FMOC-Cl. PMID- 16310418 TI - Direct determination of the ratio of tetrahydrocortisol+allo-tetrahydrocortisol to tetrahydrocortisone in urine by LC-MS-MS. AB - The 11beta-hydroxysteroid dehydrogenase (11beta-HSD) is responsible for the interconversion of both the hormonally inactive cortisone and the active cortisol. This enzyme activity, which has implications in the pathogenesis of numerous diseases, is reflected in the ratio of tetrahydrometabolites of cortisol (allo-tetrahydrocortisol and tetrahydrocortisol) to those of cortisone (tetrahydrocortisone). Several methods have been proposed in the literature to determine such a ratio in urine. Most of them require tedious and extensive extraction and derivatization steps and make use of gas-chromatographic techniques, including gas chromatography coupled to mass spectrometry (GC-MS). We present here an alternative approach for the direct determination of such a ratio in urine by using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS), based on a minimal sample treatment. Actually, the limit of detections (LODs) for pure standards in water permitted a simple dilution of the urine samples prior to the analysis, hence, an accurate optimization of the high performance liquid chromatography (HPLC) separation was needed in order to get rid of the severe influence of the urine matrix on the ionization efficiency. Besides, the nature of some interfering species was deeply investigated, as well as the suitability of some commercial deuterated steroids as internal standards. All these led to the final method, which was based on a HPLC separation on a C8 column and a ternary gradient water/methanol/acetonitrile. In parallel, an appropriate sample preparation was set up, which consisted of an enzymatic hydrolysis of the conjugated species and a followed 1:20 dilution. Preliminary measurements on real urine samples were performed as well. PMID- 16310419 TI - Standard line slopes as a measure of a relative matrix effect in quantitative HPLC-MS bioanalysis. AB - A simple experimental approach for studying and identifying the relative matrix effect (for example "plasma-to-plasma" and/or "urine-to-urine") in quantitative analyses by HPLC-MS/MS is described. Using as a database a large number of examples of methods developed in recent years in our laboratories, the relationship between the precision of standard line slopes constructed in five different lots of a biofluid (for example plasma) and the reliability of determination of concentration of an analyte in a particular plasma lot (or subject) was examined. In addition, the precision of standard line slopes was compared when stable isotope-labeled analytes versus analogs were used as internal standards (IS). Also, in some cases, a direct comparison of standard line slopes was made when different HPLC-MS interfaces (APCI versus ESI) were used for the assay of the same compound, using the same IS and the same sample preparation and chromatographic separation conditions. In selected cases, the precision of standard line slopes in five different lots of a biofluid was compared with precision values determined five times in a single lot. The results of these studies indicated that the variability of standard line slopes in different lots of a biofluid [precision of standard line slopes expressed as coefficient of variation, CV (%)] may serve as a good indicator of a relative matrix effect and, it is suggested, this precision value should not exceed 3-4% for the method to be considered reliable and free from the relative matrix effect liability. Based on the results presented, in order to assess the relative matrix effect in bioanalytical methods, it is recommended to perform assay precision and accuracy determination in five different lots of a biofluid, instead of repeat (n=5) analysis in the same, single biofluid lot, calculate standard line slopes and precision of these slopes, and to use <3-4% slope precision value as a guide for method applicability to support clinical studies. It was also demonstrated that when stable isotope-labeled analytes were used as internal standards, the precision of standard line slopes in five different lots of a biofluid was 169.99), CG (252.12-->195.02) and 4CPB (212.06- >153.06) was monitored using selected reaction monitoring. The three compounds and the internal standard (chloroproguanil) were well separated by HPLC and no interfering peaks were detected at the usual concentrations found in blood and plasma. The limit of quantification of PG and CG was 1 ng/mL and 5 ng/mL for 4CPB in rat blood and plasma. The extraction efficiency of PG, CG and 4CPB from rat blood and plasma was higher than 73%. The intra- and inter-assay variability of PG, CG and 4CPB were within 12% and the accuracy within +/-5%. This new assay offers higher sensitivity and a much shorter run time over earlier methods. PMID- 16310421 TI - Detection and quantification of ginsenoside Re in ginseng samples by a chromatographic immunostaining method using monoclonal antibody against ginsenoside Re. AB - A chromatographic immunostaining method has been developed for the determination of ginsenoside Re (G-Re) in ginseng samples on a polyethersulphone (PES) membrane. G-Re standard and the extracts of ginseng roots were applied to a PES membrane and developed by methanol-water-acetic acid (45:55:1, by volume). G-Re was clearly detected by an immunostaining method using a monoclonal antibody against G-Re. The coloring spots of G-Re were analyzed quantitatively using NIH Image software indicating at least 0.125 microg of G-Re was detectable. G-Re can be analyzed quantitatively between 0.25 and 4.0 microg. PMID- 16310422 TI - Three-dimensional domain-swapped oligomers of ribonuclease A: identification of a fifth tetramer, pentamers and hexamers, and detection of trace heptameric, octameric and nonameric species. AB - By lyophilization from 40% acetic acid solutions, bovine pancreatic Ribonuclease A forms three-dimensional domain-swapped dimers, trimers, and tetramers that can be separated by cation-exchange chromatography. Each oligomeric species consists of at least two conformers, one less basic, one more basic. The structures of the two dimers and one trimer have been solved. Plausible models have been proposed for a second RNase A trimer and four tetramers. This work is focused on the characterization of the largest oligomers which compose small peaks that have always appeared in chromatograms of RNase A. These higher order oligomers were collected by repeated cation-exchange chromatographies. On the basis of (a) gel filtrations through analytical Superdex 75 and 200; (b) gel electrophoreses under non-denaturing conditions, (c) cross-linking with divynilsulfone followed by analyses with SDS-PAGE and mass spectrometry, (d) enzymatic activity assays, and (e) analyses of the products of spontaneous dissociation of the oligomers, we could identify three-dimensional domain-swapped pentamers and hexamers, and one additional tetrameric conformer. For the latter we propose a cyclic model (C(TT)). Moreover, we advance a linear model (NCNC(P)) for one pentamer, and three possible cyclic models (with a C-trimer as the main component) for one hexamer. The experimental evidence also indicates the existence of heptameric, octameric and nonameric species. PMID- 16310423 TI - Protection, promotion and support of breastfeeding in low-income countries. AB - The rates of exclusive breastfeeding and the duration of breastfeeding fall short of what is recommended by the Global Strategy on Infant and Young Child Feeding worldwide. In low-income countries this is associated with a great excess of avoidable childhood death and disease. A higher degree of protection, promotion and support of breastfeeding has the potential to avert the death of about 1.3 million children per year and to prevent much of the associated individual and social sufferings. This paper presents some evidence about interventions that are effective to protect, promote and support breastfeeding in the health system and in the community. These interventions should not be implemented in isolation, but as part of an integrated and intersectoral programme, with a participatory approach that takes local cultural characteristics into account. Lack of political will is probably the most important factor associated with inadequate protection, promotion and support of breastfeeding. PMID- 16310424 TI - Does diabetes mellitus abolish the beneficial effect of primary coronary angioplasty on long-term risk of reinfarction after acute ST-segment elevation myocardial infarction compared with fibrinolysis? (A DANAMI-2 substudy). AB - Little is known about the effect of diabetes mellitus on long-term clinical outcome after primary percutaneous coronary intervention (pPCI) compared with fibrinolysis in patients who have acute ST-elevation myocardial infarction. We analyzed 3-year clinical outcome in diabetic patients and nondiabetic patients who had been randomized to fibrinolysis or pPCI in the DANAMI-2 trial to compare long-term clinical outcome. The primary end point was a composite of death, clinical reinfarction, or disabling stroke. Median follow-up was 3.8 years. Among 1,572 consecutive patients who had ST-elevation myocardial infarction and were randomized to pPCI or fibrinolysis, 173 (11.0%) had diabetes mellitus; 60 of these patients received metformin treatment and were excluded. After 3 years no difference was found between diabetic patients who underwent pPCI versus fibrinolysis (combined event p=0.37, reinfarction p=0.06 in favor of fibrinolysis), whereas pPCI was superior to fibrinolysis in nondiabetic patients (combined event p=0.002, clinical reinfarction p<0.001). Three-year incidence of clinical reinfarction analyzed with Cox's regression showed that pPCI compared with fibrinolysis increased the relative risk of clinical reinfarction in diabetic patients (relative risk 2.57, 95% confidence interval 1.48 to 4.46, p <0.001) but decreased the risk in nondiabetic patients (relative risk 0.52, 95% confidence interval 0.36 to 0.74, p<0.001). In conclusion, from the DANAMI-2 trial we hypothesize that diabetes may abolish the beneficial effect of pPCI on long-term risk of clinical reinfarction. PMID- 16310425 TI - Association of plaque characterization by intravascular ultrasound virtual histology and arterial remodeling. AB - Positive remodeling is more often observed in lesions of patients who have acute coronary syndromes or vulnerable (rupture-prone) plaques. However, there are few data that correlate plaque morphology, composition, and arterial remodeling in vivo. We evaluated coronary plaque characterization of lesions with positive remodeling using intravascular ultrasound (IVUS) radiofrequency data analysis. Seventy-seven nonbifurcation native coronary lesions (in 50 patients) were imaged in vivo using 30-MHz IVUS transducers. Lesions were classified into 4 plaque types, fibrous, fibrofatty, dense calcium, and necrotic core, by using processing of the radiofrequency signal validated in vitro. The remodeling index was calculated as the lesion external elastic membrane area divided by the proximal reference external elastic membrane area. Lesions were divided into 2 groups: positive remodeling (remodeling index>1.0, 26 lesions) and intermediate/negative remodeling (remodeling index60 minutes could be justified for high-risk patients. PMID- 16310431 TI - Relation of low socioeconomic status to C-reactive protein in patients with coronary heart disease (from the heart and soul study). AB - Low socioeconomic status (SES) is associated with poor health outcomes in patients who have coronary heart disease (CHD). Inflammation is a potential mechanism by which low SES may lead to adverse cardiovascular outcomes, but it is not known whether low SES is associated with inflammation in patients who have CHD. We measured high-sensitivity C-reactive protein (CRP) levels in a cross sectional study of 985 adults who had CHD. Income and education were determined by self-report. We used ordinal logistic regression to examine the association of income and education with CRP. Of the 985 participants, 390 had high CRP levels (>3 mg/dl). The proportion of participants who had high CRP levels ranged from 30% (103 of 340) in those who had a college degree to 51% (65 of 127) in those who had less than a high school degree (p<0.0001). The proportion of subjects who had a high CRP level ranged from 28% (52 of 183) in those who had annual income>or=$50,000 to 42% (199 of 974) in those who had an annual income<$20,000 (p<0.001). After adjustment for traditional cardiovascular risk factors and other potential confounding variables, lower income and education remained associated with higher CRP levels. In conclusion, low SES is associated with high CRP levels in patients who have CHD. This observation raises the possibility that inflammation may contribute to the adverse cardiovascular outcomes associated with low SES. PMID- 16310432 TI - Frequency of distress and fear of dying during acute coronary syndromes and consequences for adaptation. AB - Experiencing an acute coronary syndrome (ACS) may provoke a range of negative emotional responses, including acute distress and fear of dying. The frequency of these emotional states has rarely been assessed. This study examined the presence and severity of the fear of dying and acute distress in 184 patients with ACS and analyzed its correlates and consequences. Intense distress and fear of dying was reported by 40 patients (21.7%) and moderate fear and distress by 95 patients (51.6%). Intense distress and fear was associated with female gender (odds ratio [OR] 2.49, 95% confidence interval [CI] 1.07 to 2.49), lower levels of education (OR 2.44, 95% CI 1.02 to 5.87), greater chest pain (OR 5.33, 95% CI 1.40 to 20.4), and emotional upset in the 2 hours before onset of ACS (OR 2.70, 95% CI 1.13 to 6.45). Having no acute distress or fear was more common in patients who exercised regularly (OR 3.32, 95% CI 1.35 to 8.18) and who did not initially attribute the chest pain to cardiac causes (OR 2.67, 95% CI 1.10 to 6.47). No association was found with cardiovascular disease history, objective measures of clinical severity, or with clinical presentation of ACS. Acute distress and fear of dying predicted greater depression and anxiety 1 week after ACS (p=0.006), and elevated levels of depression at 3 months (p=0.009), after adjustment for age, gender, and negative affect. In conclusion, distress and fear during the initial stages of an ACS may trigger subsequent depression and anxiety, thereby promoting poorer prognosis and greater morbidity with time. PMID- 16310433 TI - Effect of glucose-insulin-potassium infusion on myocardial damage due to percutaneous coronary revascularization. AB - Percutaneous coronary intervention has been known to cause myocardial damage as a result of microvascular dysfunction due to microembolization and microinfarction. Previous studies have shown that glucose-insulin-potassium (GIK) infusion decreases mortality in patients with acute myocardial infarction. Therefore, in this study, we aimed to investigate the effect of GIK infusion on myocardial damage due to percutaneous coronary revascularization. A total of 52 consecutive nondiabetic patients diagnosed with non-ST-elevation acute coronary syndrome and designated for elective percutaneous coronary intervention were randomized in a double-blind fashion into GIK and normal saline groups. GIK infusion (30% dextrose, 300 U insulin, and 60 mEq potassium chloride) at a dose of 1.5 ml/kg/hour was initiated 24 hours before the intervention and continuing during and until 1 hour after the intervention. Troponin I levels were recorded in venous blood samples before and 12 and 24 hours after the intervention. The increase in troponin I was significantly lower at 12 and 24 hours in the GIK group compared with those of the saline controls (p=0.022 and p=0.005, respectively). GIK infusion initiated 24 hours before coronary stenting for non ST-elevation acute coronary syndrome resulted in less myocardial damage as determined by postprocedure troponin I levels. PMID- 16310434 TI - Usefulness of omega-3 fatty acids and the prevention of coronary heart disease. AB - Clinical trial evidence exists that supports a role for the omega-3 polyunsaturated fatty acids in coronary heart disease prevention. However, the results from these clinical trials have varied and were conducted in diverse population groups using several different types of omega-3 polyunsaturated fatty acids, including eicosapentaenoic acid, docosahexaenoic acid, and alpha-linolenic acid (ALA). Thus, we systematically reviewed previously published reports assessing the different types of omega-3 polyunsaturated fatty acid interventions and cardiovascular outcomes. Fourteen randomized clinical trials were included in the review. Six trials were included with fish oil, with 1 large trial (10,000 patients) dominating the analysis. In aggregate, the fish oil trials demonstrated a reduction in total mortality and sudden death without a clinically significant reduction in nonfatal myocardial infarction. The 6 trials with ALA supplements or an ALA-enriched diet were of poorer design than the fish oil trials and had limited power. Many of the trials with ALA involved other changes in dietary components. In aggregate, the ALA trials demonstrated possible benefits in reducing sudden death and nonfatal myocardial infarction, but with wider confidence intervals than in the fish oil trials. In conclusion, the evidence suggests a role for fish oil (eicosapentaenoic acid, docosahexaenoic acid) or fish in secondary prevention because recent clinical trial data have demonstrated a significant reduction in total mortality, coronary heart disease death, and sudden death. The data on ALA have been limited by studies of smaller sample size and limited quality. PMID- 16310435 TI - Hospitalizations for new heart failure among subjects with diabetes mellitus in the RENAAL and LIFE studies. AB - We sought to study the risk factors for heart failure (HF) and the relation between antihypertensive treatment with losartan and the first hospitalization for HF in patients with diabetes mellitus in the Losartan Intervention For Endpoint reduction in hypertension (LIFE) and Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) studies. We evaluated 1,195 patients with hypertension, left ventricular hypertrophy, and diabetes from the LIFE study and 1,513 patients with type 2 diabetes and nephropathy from the RENAAL study. The comparative treatments were atenolol in the LIFE study and placebo in the RENAAL study. Patients with a history of HF were excluded from this analysis. Losartan significantly reduced the incidence of first hospitalizations for HF versus placebo in the RENAAL study (hazard ratio 0.74, p=0.037) and versus atenolol in the LIFE study (hazard ratio 0.57, p=0.019). Patients enrolled in the RENAAL study were at a higher risk of developing HF (hazard ratio for RENAAL vs LIFE diabetics 3.0, p<0.0001). The significant, independent baseline risk factors for the development of HF in the RENAAL study were urinary albumin/creatinine ratio, age, peripheral vascular disease, the Cornell product, body mass index, and previous angina; in the LIFE study they were the Cornell product, previous myocardial infarction, peripheral vascular disease, baseline atrial fibrillation, alcohol use (inverse relation), and urinary albumin/creatinine ratio. The beneficial effect of losartan on the reduction of risk for hospitalization for new HF was demonstrated in patients who were at high renal and/or high cardiovascular risk. PMID- 16310436 TI - United States emergency department visits for acute decompensated heart failure, 1992 to 2001. AB - Heart failure is a significant public health problem. The epidemiology and practice pattern of emergency department (ED) visits for acute decompensated heart failure (ADHF) have not been well characterized. A better description is essential to highlight areas in which improvements or additional research are needed. We analyzed all ED visits for ADHF of patients>or=40 years old using the data of the National Hospital Ambulatory Medical Care Survey from 1992 to 2001. During this 10-year period, an estimated 10.5 million ED visits occurred for ADHF, representing 2.9% of all ED visits. The number of ADHF visits increased on average by 18,500 per year, for a 19.4% absolute increase during the decade. The rate per 1,000 United States population was unchanged. The average patient was 74 years old, and patients>or=65 years accounted for 79% of visits. Loop diuretics were administered in 63% and vasodilators in 29% of visits. The ED visit rate per 1,000 United States population was 53% higher in blacks than in whites (14.2 vs 9.3). In a multivariate model, white race was a significant predictor of hospitalization. In conclusion, during the past decade, the absolute number of ED visits for ADHF has increased owing to the aging population. Diuretics remain the most common treatment. Race-related differences in hospitalization merit additional study. PMID- 16310437 TI - Prevalence of mechanical dyssynchrony in patients with heart failure and preserved left ventricular function (a report from the Belgian Multicenter Registry on dyssynchrony). AB - The present study evaluated the prevalence of mechanical inter- and intraventricular dyssynchrony in patients with heart failure and preserved left ventricular (LV) ejection fraction (LVEF). We studied 138 patients with heart failure (age 67+/-11 years; 76% men); 60 patients had preserved LVEF (>40%). Using conventional Doppler echocardiography, an interventricular mechanical delay>or=40 ms was defined as interventricular dyssynchrony. Using pulse-wave tissue Doppler imaging, the time from the beginning of the QRS complex to onset of systolic motion was measured in 4 basal LV segments. A dispersion of >or=60 ms was defined as intraventricular dyssynchrony. The prevalence of inter- and intraventricular dyssynchrony was lower in patients with preserved LVEF than in those with reduced LVEF (17% vs 41%, p<0.01 for interventricular dyssynchrony, 18% vs 36%, p<0.01 for intraventricular dyssynchrony). However, patients with preserved LVEF and a QRS width>or=120 ms had higher values for the parameters for inter- and intraventricular dyssynchrony than patients with a QRS width<120 ms (interventricular mechanical delay 33+/-20 vs 20+/-16 ms, p<0.05; tissue Doppler imaging dispersion 42+/-26 vs 33+/-22 ms, p<0.05). In patients with a QRS width>or=120 ms, the prevalence of inter- and intraventricular dyssynchrony was comparable for patients with preserved and reduced LVEF (42% vs 55%, p=NS for interventricular dyssynchrony and 45% vs 46%, p=NS for intraventricular dyssynchrony). In conclusion, the prevalence of inter- and intraventricular dyssynchrony was low (17% and 18%, respectively) in patients with heart failure and preserved LVEF. However, in the presence of a QRS width of >or=120 ms, this prevalence increased to almost 50%, comparable to that for patients with heart failure and reduced LVEF and a QRS width of >or=120 ms. PMID- 16310438 TI - Comparison of diameter of ascending aorta in patients with severe aortic stenosis secondary to congenital versus degenerative versus rheumatic etiologies. AB - Aortic root dilation has been previously reported to be associated with aortic stenosis (AS), but data to support this statement are scarce. The dimensions of the aortic root were measured at 4 levels (annulus, sinuses, sinotubular junction, and ascending aorta) in 88 patients (mean age 71.2+/-9.7 years; 56% men) with severe AS who underwent intraoperative transesophageal echocardiography immediately before aortic valve replacement. These patients were compared with 76 gender- and age-matched patients without AS who underwent transesophageal echocardiography for various indications. The etiology of aortic valve stenosis was degenerative in 62 (70.5%), bicuspid aortic valve (BAV) in 15 (17.0%), and rheumatic in 11 (12.5%). The ascending aorta was significantly wider in AS with various etiologies (BAV, rheumatic, degenerative) than in the controls (39+/-6.9, 35.0+/-4.2, 33.1+/-4.1, and 31.3+/-3.7 mm, respectively; p<0.001). The dimensions of the sinuses and sinotubular junction were significantly less in those with AS of degenerative etiology than in the controls (29.5+/-4.0 vs 32.5+/-4.3 mm and 23.6+/-3.0 vs 26.8+/-3.0 mm, respectively, p<0.001). The prevalence of a dilated aorta (>37 mm) was 3.9%, 13.1%, 36.4%, and 60% in the control group and AS patients with degenerative, rheumatic, and BAV etiology, respectively (p<0.0001). In conclusion, patients with severe AS due to BAV had significant dilation of the aortic root. Patients with degenerative and rheumatic etiology had less remarkable dilation compared with control group, and most values were within the normal range. PMID- 16310439 TI - Clinical determinants of sinus conversion by radiofrequency maze procedure for persistent atrial fibrillation in patients undergoing concomitant mitral valvular surgery. AB - The radiofrequency Maze procedure can effectively restore sinus rhythm in most patients with atrial fibrillation (AF) and mitral valve disease. AF after cardiac surgery is associated with increased morbidity and mortality. However, clinical determinants of long-term postoperative AF after the radiofrequency Maze procedure and concomitant mitral valve surgery are poorly defined. This study comprised 99 consecutive patients with persistent AF and mitral valve disease who underwent radiofrequency Maze procedures and concomitant mitral valvular operations. The predictive values of clinical variables for postoperative AF were examined. After a mean follow-up period of 46.1+/-24.6 months, 83 patients (83.8%) had sinus conversion after the Maze procedure, and 16 patients remained in persistent or paroxysmal AF. Multiple logistic regression analysis determined that predictors of sinus conversion were preoperative left atrial diameter (odds ratio [OR] 1.127 per 1-mm increment in left atrial diameter, 95% confidence interval [CI] 1.045 to 1.215, p<0.002) and the duration of AF (OR 1.022 per 1 month increment in duration of AF, 95% CI 1.009 to 1.035, p<0.001). Discriminant analysis showed that the sinus conversion rate was significantly lower in patients with preoperative left atrial diameters>56.8 mm (p<0.001) or AF duration>66 months (p<0.001) than in patients with preoperative left atrial diameters<56.8 mm or AF duration<66 months. In conclusion, the preoperative left atrial size and duration of AF are primary predictors of sinus conversion by the radiofrequency Maze procedure for patients with persistent AF and mitral valve disease. PMID- 16310440 TI - Usefulness of tissue Doppler imaging for demonstrating altered septal contraction sequence during dual-chamber pacing in obstructive hypertrophic cardiomyopathy. AB - Dual-chamber pacing reduces left ventricular (LV) outflow obstruction in patients with obstructive hypertrophic cardiomyopathy (HC), the mechanism of which lies in pacing-induced paradoxic septal motion. This study was conducted to test the hypothesis that tissue Doppler imaging (TDI) could demonstrate changes in the septal contraction sequence during dual-chamber pacing in patients with HC. TDI was performed in 16 patients (5 women; mean age 63+/-11 years) who underwent dual chamber pacing for 7.6+/-2.1 year. With and without pacing, the time to peak systolic myocardial velocity was measured from the basal, mid, and distal segments in the 4 different LV walls. Without pacing, there was almost no longitudinal segmental asynchrony. During pacing, however, marked longitudinal segmental asynchrony appeared, especially in the anteroseptal wall (from p=NS to p<0.01 by analysis of variance) and the ventricular septum (from p<0.05 to p<0.01), with the time to peak velocity extremely prolonged at the distal segments. This was associated with a modest but significant decrease in the LV pressure gradient (from 20+/-8 to 14+/-7 mm Hg, p<0.01). In patients with obstructive HC, altered septal contraction sequence accounts for the reduced LV outflow obstruction during dual-chamber pacing, which was clearly demonstrated by TDI. PMID- 16310441 TI - Effect of Losartan on left ventricular diastolic function in patients with nonobstructive hypertrophic cardiomyopathy. AB - In hypertrophic cardiomyopathy (HC), diastolic dysfunction of the left ventricle is a prominent feature caused by myocardial hypertrophy and fibrosis. Angiotensin II has trophic and profibrotic effects on the heart, and the blockade of angiotensin II receptors reverses hypertrophy and fibrosis in human cardiac diseases and in animal HC. This study investigated the short-term (6 months) effects of losartan 100 mg/day in 20 patients with nonobstructive HC, with an emphasis on left ventricular (LV) diastolic dysfunction, compared with 10 patients with HC who were not treated. At the final evaluation, significant changes were observed in the losartan group: a left atrial diameter decrease (p<0.0001), a tissue Doppler early (Ea) mitral annulus diastolic velocity increase (p=0.003) and an E/Ea ratio decrease (p=0.0002), and a significant decrease in plasma levels of the aminoterminal fragment of pro-brain natriuretic peptide (NT-pro-BNP) from a median of 860 to 606 pg/ml (p=0.001). A significant correlation was found between percentage changes in NT-pro-BNP and the E/Ea ratio from baseline to 6 months (r=0.61, p=0.002). In the 2 groups, echocardiographic LV wall and cavity measures did not change. In conclusion, in selected patients with nonobstructive HC, losartan during a 6-month period improved LV diastolic function. PMID- 16310442 TI - Usefulness of magnetic resonance imaging of left ventricular endocardial fibroelastosis in infants after fetal intervention for aortic valve stenosis. AB - This report describes the use of myocardial delayed-enhancement magnetic resonance imaging to delineate the extent of endocardial fibroelastosis (EFE) in 4 infants after fetal balloon aortic valvuloplasty. The preoperative mapping of EFE followed by extensive resection led to biventricular physiology in 2 patients. PMID- 16310443 TI - Systemic oxygen saturation and coagulation factor abnormalities before and after the fontan procedure. AB - Coagulation factor abnormalities are believed to predispose to increased risk for thromboembolism after the Fontan procedure. Recent data, however, suggest that coagulation abnormalities may precede the operation. This study determined coagulation factor abnormalities in patients with single-ventricle congenital heart defects before the Fontan procedure and compared the findings with age matched post-Fontan patients and control subjects. Liver function, coagulation factor levels, and pulse oximetry readings were determined in 11 pre-Fontan patients aged 9.3+/-5.6 years. The results were compared with those of 11 age matched post-Fontan patients, at 9.1+/-2.4 years after the Fontan procedure, and 11 age-matched controls by simple analysis of variance and post hoc Tukey's test. Liver function was normal in patients before and after surgery apart from mildly elevated bilirubin levels in post-Fontan patients (p=0.027). When compared with controls, pre-Fontan patients had significantly lower levels of protein C; protein S; antithrombin III; and factors II, V, VII, and X and longer prothrombin times (all with p<0.05). When compared with post-Fontan patients, pre-Fontan patients had similarly lower levels of free protein S (p<0.001), protein C (p=0.06), and antithrombin III (p=0.001). Systemic oxygen saturation correlated positively with the levels of anticoagulants (protein C, protein S, antithrombin III) and procoagulants (factors II, V, VII, and X) (all with p<0.05, r=0.33 to 0.61). In conclusion, the findings suggest that coagulation factor abnormalities precede the Fontan procedure and tend to normalize after the operation, relating perhaps to improved systemic oxygenation. PMID- 16310444 TI - Status of endothelial dependent vasodilation in patients with hyperuricemia. AB - Hyperuricemia has been associated with an increased risk for cardiovascular disease and increased mortality. However, the biologic mechanisms that link elevated serum uric acid to cardiovascular disease are uncertain. This study tested the hypothesis that elevated serum uric acid is associated with impaired endothelial function in hyperuricemic patients without any overt cardiovascular disease. Seventeen male patients with hyperuricemia (mean age 42+/-4 years) and 9 control subjects (mean age 45+/-5 years) were studied. All subjects were nonsmokers. All patients had never been treated for hyperuricemia, were on no medications, and were free of any other known diseases. Endothelial function was evaluated by flow-mediated dilation measured by ultrasound. Flow-mediated dilation was significantly impaired in patients with hyperuricemia (4.0+/-0.7%) compared with control subjects (6.4+/-0.8%) (p=0.044). Flow-mediated dilation correlated inversely with uric acid levels (r=-0.4, p=0.05). Nitrate-induced dilation was 12.3+/-1.0% in patients with hyperuricemia and 11.8+/-2.3% in control subjects (p=0.82). Impaired endothelial-dependent vasodilation is present in hyperuricemic patients even in the absence of any overt cardiovascular disease. The elevated serum uric acid, per se, may constitute a novel risk factor for endothelial dysfunction. PMID- 16310445 TI - Relation of inflammation to peripheral arterial disease in the national health and nutrition examination survey, 1999-2002. AB - The relation between inflammation and peripheral arterial disease (PAD) is not well characterized. This study examined this relation and its consistency across important subgroups in a cross-sectional, nationally representative sample of the adult United States population. C-reactive protein (CRP), fibrinogen, leukocyte count, and PAD were assessed in a sample of 4,787 participants aged>or=40 years in the National Health and Nutrition Examination Survey 1999-2002. PAD was defined as an ankle-brachial blood pressure index <0.9. Graded relations were present between inflammatory markers and PAD. The multivariate adjusted odds ratios of PAD associated with the highest versus the lowest quartile of CRP, fibrinogen, and leukocyte count were 2.14 (95% confidence interval [CI] 1.41 to 3.25), 2.49 (95% CI 1.27 to 4.85), and 1.67 (95% CI 0.84 to 3.31), respectively (each p trend<0.05 across quartiles). Associations between inflammation and PAD were similar across gender, obesity, and diabetic subgroups. However, the odds ratios of PAD for the highest CRP quartile versus the 3 lowest quartiles were 3.10 (95% CI 1.76 to 5.45) for non-Hispanic blacks versus 1.50 (95% CI 0.98 to 2.28) for non-Hispanic whites and 1.11 (95% CI 0.57 to 2.17) for Mexican Americans (p interaction=0.049) and 5.59 (95% CI 1.82 to 17.17) for patients aged 40 to 54 years versus 2.01 (95% CI 1.13 to 3.58) for patients aged 55 to 69 years and 0.98 (95% CI 0.65 to 1.48) for patients aged>or=70 years (p interaction=0.018). Odds ratios of PAD for the highest fibrinogen quartile versus the lowest 3 quartiles were 3.26 (95% CI 1.69 to 6.28) for current smokers versus 0.83 (95% CI 0.51 to 1.35) for never smokers (p interaction=0.006). In conclusion, in the general United States adult population, inflammation is independently associated with PAD. PMID- 16310446 TI - Comparative accuracy of manual versus computerized electrocardiographic measurement of J-, ST- and T-wave deviations in patients with acute coronary syndrome. AB - Accurate and rapid electrocardiographic interpretation is of crucial importance in acute coronary syndrome (ACS). Computerized electrocardiographic algorithms are often used in out-of-hospital settings. Their accuracy should be carefully validated in ACS, particularly in ST-elevation myocardial infarction. This study evaluated the comparative accuracy of lead-specific computer-based versus manual measurements of the J-point, ST-segment, and T-wave deviations in standard 12 lead electrocardiograms (ECGs) (excluding lead aVR). Sixty-nine consecutive patients with suspected ACS were included. The interobserver reliability in the determination of ST-segment deviation>or=0.2 mV in leads V2 and V3 was very good (kappa=0.94 and 0.93, respectively). Agreement between a cardiologist and the computer regarding ST elevation>or=0.2 mV in lead V2 was moderate (kappa=0.72) and in V3 was very good (kappa=0.85). For ST depression or elevation>or=0.05 mV in lead LIII, agreement was good and moderate (kappa=0.79 and 0.51, respectively). Bland-Altman analysis demonstrated clinically acceptable limits of agreement comparing measurements of the J point and the T wave, but clinically inadequate limits of agreement with respect to ST-segment deviation, between the electrocardiographer and the computer. The optimal cut-off points were 0.115 mV (sensitivity 89%, specificity 98%) for the computer program to detect ST elevation>or=0.2 mV and 0.045 mV (sensitivity 74%, specificity 99%) for revealing ST elevation>or=0.1 mV. It was found that automatically measured ST-segment deviations were smaller than those manually measured. In conclusion, a correction should be performed to obtain optimal results in the automated analysis of ECGs, because the results have important implications for clinical decision making. PMID- 16310447 TI - The Editor's roundtable: cyclooxygenase-2 inhibitors and cardiovascular risk. PMID- 16310448 TI - Aorta to right atrial fistula following transcatheter closure of an atrial septal defect. AB - The percutaneous transcatheter closure of atrial septal defects (ASDs) has become an acceptable alternative to surgical repair. The investigators present a case of aorta-to-right atrial fistula after the closure of an ASD using the Amplatzer septal occluder (ASO). After the surgical removal of the ASO, the ASD patch closure, and the repair of the fistula, the patient eventually recovered. PMID- 16310449 TI - Migration of an amplatzer septal occluder device for closure of atrial septal defect into the ascending aorta with formation of an aorta-to-right atrial fistula. AB - The percutaneous closure of atrial septal defects is increasingly used. Serious complications of the procedure, such as cardiac perforation and tamponade, are rare and usually occur <72 hours after device placement. The investigators report the late development of the erosion of an Amplatzer septal occluder into the ascending aorta with associated aortic-to-right atrial fistula formation. PMID- 16310450 TI - Systemic hypertension: some observations. PMID- 16310451 TI - Is vasospastic angina an inflammatory disease? PMID- 16310452 TI - An association between prinzmetal's angina pectoris and obstructive lung disease. PMID- 16310453 TI - What makes Lance Armstrong tick? He has a big ticker. PMID- 16310454 TI - A unique eponymous sign of finger clubbing (Schamroth sign) that is named not only after a physician who described it but also after the patient who happened to be the physician himself. PMID- 16310455 TI - Metabolic syndrome, hostility, and cardiac rehabilitation. PMID- 16310456 TI - Tractography to depict three layers of visual field trajectories to the calcarine gyri. AB - PURPOSE: Fiber-tracking by diffusion tensor magnetic resonance imaging (DT-MRI) is currently the only noninvasive in vivo method for white matter fiber-tracking in the human brain. We used this method in attempts to visualize the optic radiation and to examine the clinical applicability of this technique. DESIGN: Observational case series. METHODS: DT-MRI scans for fiber-tracking were obtained in five healthy volunteers by use of a whole-body, 1.5 Tesla imager. DT-MRI data were transferred to an off-line workstation; PRIDE software was used for image analysis. We constructed 3 diopters fiber trajectories by tracking the direction of the fastest diffusion from the lateral geniculate nucleus, and then selected tracts on the basis of anatomical knowledge of the optic radiation. RESULTS: Our method successfully reconstructed the macroscopic 3 diopters architecture of the three major groups of optic radiation in all subjects. Meyer's loop depicted by tractography was located more posterior than the known anatomical locations, although our results on the central and posterior bundles were in good agreement with them. DT-MRI scanning required 7 minutes; preliminary images of the optic radiation could be obtained in approximately 20 minutes. CONCLUSIONS: Fiber tracking enabled us to obtain information quickly on the 3 diopters course of the optic radiation in vivo. The finding that the fiber-tracking method underestimates the anterior extent of the optic radiations could prove to be an important limitation in the utility of this technique for preoperative planning. The time required for data acquisition and processing makes this method acceptable for routine clinical use. PMID- 16310457 TI - Multifocal electroretinographic changes in patients receiving hydroxychloroquine therapy. AB - PURPOSE: To evaluate the longitudinal changes in multifocal electroretinography (mfERG) in patients receiving hydroxychloroquine and to assess the effects of cumulative hydroxychloroquine dose on mfERG. DESIGN: Prospective cohort study. METHODS: Twenty-four eyes in 12 patients receiving hydroxychloroquine underwent mfERG recordings at baseline and 1 to 2 years later. The first negative (N1) and first positive (P1) response amplitudes and peak latencies were compared with normal controls. Serial changes in the pattern of mfERG abnormalities and in response amplitudes and peak latencies were also compared between eyes in which hydroxychloroquine therapy was continued or stopped. Correlation analyses were performed to assess the effects of a cumulative dose of hydroxychloroquine on mfERG. RESULTS: At baseline, reductions in N1 and P1 response amplitudes were observed in patients receiving hydroxychloroquine compared with controls. At follow-up, in addition to the reductions in N1 and P1 response amplitudes, increases in P1 peak latencies compared with controls were observed. In patients who stopped hydroxychloroquine therapy, there were significant increases in N1 and P1 response amplitudes at follow-up mfERG. CONCLUSIONS: Patients receiving hydroxychloroquine showed a longitudinal decline in retinal function; patients who stopped hydroxychloroquine therapy showed improvement. Although these data are insufficient to demonstrate the sensitivity of mfERG for evaluating early hydroxychloroquine toxicity, the results suggest that serial mfERG assessment may help detect early retinal changes associated with hydroxychloroquine therapy. Further studies with long-term results will be useful in clarifying the value of mfERG in evaluating early retinal toxicity due to hydroxychloroquine. PMID- 16310458 TI - Screening for diabetic retinopathy by ophthalmologists and endocrinologists with pupillary dilation and a nonmydriatic digital camera. AB - PURPOSE: To assess the effects of pupillary dilation on image quality and certitude of screening diabetic retinopathy by trained endocrinologists using a digital nonmydriatic camera. DESIGN: Prospective, comparative, observational case series. METHODS: One hundred fifty patients with diabetes attending a hospital based photographic screening clinic had five-field (45 degree) digital retinal imaging and mosaic construction, first through dark-adapted pupils, then after a single application of tropicamide 1%. The 600 sets of images (150 patients, 300 eyes, before and after dilation) were graded independently and in a blinded manner by two endocrinologists and two ophthalmologists, with the consensual opinion on dilated images of the latter serving as the gold standard. The criteria of evaluation were assessment of image quality and certitude of detecting and grading retinopathy. RESULTS: Of 300 eyes, pharmacological mydriasis improved image quality, with an increase in the number of eyes with five good images from seven to 160 and good-quality mosaics from 99 to 233. Dilation allowed better identification of maculopathy in the second eye because there was a decrease in ungradeable central images from 127 to 15 eyes. Dilation increased the number of eyes having retinopathy detected with certainty from 153 to 252 and graded with certainty from 173 to 277. No adverse effects were recorded. CONCLUSIONS: Single application of tropicamide 1% significantly improves image quality and certitude of screening diabetic retinopathy with nonmydriatic camera by endocrinologists. PMID- 16310459 TI - Diagnostic testing of vitrectomy specimens. AB - PURPOSE: To assess the usefulness of diagnostic tests that are performed on vitrectomy specimens from patients with suspected lymphoma or infection. DESIGN: Noncomparative, retrospective, interventional case series. METHODS: Seventy-eight consecutive patients (84 eyes) underwent pars plana vitrectomy for diagnostic purposes. Vitrectomy with cytologic, cytofluorographic, or microbiologic analysis of vitreous samples was performed. The main outcome measures were the efficiency of diagnostic procedure and positive and negative predictive values (PPV and NPV). RESULTS: There were 28 patients (33 eyes) with suspected intraocular lymphoma and 50 patients (51 eyes) with suspected infection, which was subdivided into chronic endogenous endophthalmitis, atypical chorioretinitis, or chronic postoperative inflammation. Vitreous testing led to a diagnosis in 48 of 78 patients (61.5%); 14 patients with a final diagnosis of lymphoma/leukemia, and 34 patients with a final diagnosis of infection. When preoperative indication was compared with final clinical diagnosis, the efficiency of the diagnostic procedure of cytologic evaluation, flow cytometry, and bacterial/fungal culture was 67%, 79%, and 96%, respectively. For lymphoma, the PPV of cytologic evaluation was 100% and the NPV 60.9%. For infection, the PPV of bacterial/fungal culture was 100% and the NPV 94.9%. CD22+ B lymphocytes >or=20% of total cells on cytofluorographic analysis had a PPV of 88% for lymphoma. A cytofluorographic CD4:CD8 T-lymphocyte ratio >or=4 had a PPV of 70% for immunologically mediated uveitis. Surgical complications were rare and manageable. CONCLUSION: Diagnostic vitrectomy in selected patients with carefully planned testing is an effective means of supporting diagnoses in intraocular lymphoma, chronic intraocular infections, and atypical chorioretinitis. Flow cytometry quantitates the percentages and ratios of various cell types and is helpful in contrasting intraocular lymphoma with immunologically mediated uveitis. PMID- 16310460 TI - Effect of artificial tears on visual acuity. AB - PURPOSE: To study the effect of commonly used preservative free artificial tear, carboxymethylcellulose (CMC) 0.5% (Refresh Plus, Allergan, Irvine, California) on visual acuity in symptomatic dry eye (SDE) and asymptomatic dry eye (ADE) patients. DESIGN: Nonrandomized prospective clinical trial. METHODS: Prospective study involving 20 patients (40 eyes) with SDE and 20 patients (40 eyes) with ADE, all 40 years and older, were recruited from a clinic setting over a 1-month period. Distance visual acuity was measured by the Early Treatment of Diabetic Retinopathy Study (ETDRS) vision chart and near visual acuity was measured by the Lighthouse Near Vision chart before and 30 seconds after instillation of one drop of CMC. Distance and near visual acuity was measured both with and without correction. The duration of action of CMC was measured at 1-minute intervals until the patient's visual acuity returned to pretear level. RESULTS: In both SDE and ADE groups, uncorrected and corrected near and distance vision showed a statistically significant improvement after the use of CMC (P < .05). There was no statistically significant difference in improvement between the SDE and ADE groups in all categories (P values > .05). The mean duration of improvement of vision was 2.93 minutes in the SDE group and 3.70 minutes in the ADE group (P = .036). CONCLUSIONS: CMC 0.5% provides a temporary yet significant improvement in the visual acuity of SDE and ADE patients. The effect of artificial tears on visual acuity may be of diagnostic value in detecting ocular surface abnormality in symptomatic and asymptomatic patients. PMID- 16310461 TI - BCL10 expression in ocular adnexal lymphomas. AB - PURPOSE: To study BCL10 expression in ocular adnexal lymphoma in the US population and its association with clinical outcomes. DESIGN: Institutional, retrospective study. METHODS: Immunohistochemistry was performed with antibody against BCL10 on two tissue microarray blocks that were constructed with paraffin embedded tissues from the same cohort of 48 patients with ocular adnexal lymphomas. The main outcomes that were measured include extraorbital involvement, recurrence rate, and time to recurrence. The median length of the follow-up period was 40 months. RESULTS: Aberrant BCL10 expression (nuclear [moderate intensity] and cytoplasmic [weak to moderate intensity] staining) was observed in 10 of 33 cases (30.3%) of mucosa-associated lymphoid tissue (MALT) lymphoma, in 4 of 10 cases (40%) of follicular lymphoma (grade 1, 9 cases; grade 2, 1 case), in 0 of 2 cases of diffused large B-cell lymphoma, in 0 of 1 case of chronic lymphocytic leukemia/small lymphocytic lymphoma and in 1 of 1 case (100%) of mantle cell lymphoma. There were no differences in clinical parameters at examination (ie, average age, gender, site of occurrence, laterality, extraorbital involvement at diagnosis), recurrence rate, and time to recurrence for patients (MALT lymphoma or follicular lymphoma) with or without aberrant nuclear BCL10 expression. CONCLUSION: Aberrant BCL10 expression can occur in other types of ocular adnexal lymphomas besides MALT lymphoma. Ocular adnexal MALT lymphoma may have slightly lower frequency of aberrant BCL10 expression than gastric/pulmonary MALT lymphomas that have been reported in the literature. Furthermore, aberrant BCL10 nuclear expression in ocular adnexal lymphoma does not seem to correlate with clinical outcome. Further studies that include a larger number of cases and longer follow-up period are needed to confirm our observation. PMID- 16310462 TI - Prevalence of Staphylococcus epidermidis strains with biofilm-forming ability in isolates from conjunctiva and facial skin. AB - PURPOSE: To compare the prevalence of biofilm-forming strains of Staphylococcus epidermidis in the conjunctival and facial skin microflora. DESIGN: Experimental study. METHODS: The prevalence of biofilm-forming ability of 10 S epidermidis strains obtained from the conjunctival sac of healthy volunteers was compared with 40 strains obtained from the facial skin of healthy volunteers. The ability to form biofilm was determined by the presence of the icaA gene, and the production of biofilm was examined by qualitative (Congo red agar [CRA]) and quantitative (microtiter plate) assays. Additionally, the prevalence of 36 S epidermidis strains obtained from the conjunctival sac of precataract patients to form biofilm was investigated. RESULTS: The icaA gene was detected in 60% of the isolates from the conjunctival sac of volunteers and 15% of those from the facial skin. Fifty percent of the isolates from the conjunctiva of volunteers and 5% from the facial skin were CRA positive. Biofilm production was significantly greater in isolates from the conjunctiva of volunteers. Of the nine pairs of isolates found in the same volunteers, six conjunctival sac isolates were positive for the icaA gene with biofilm-forming ability except one strain, whereas only one of the facial skin isolates was positive for the icaA gene and none exhibited biofilm-forming phenotype. Sixty-nine percent and 44% of the isolates from the conjunctival sac of precataract patients were positive for icaA gene and CRA test, respectively. CONCLUSIONS: The prevalence of biofilm-forming S epidermidis isolates is higher in the conjunctival sac than the facial skin. PMID- 16310463 TI - Sensitive and rapid polymerase chain reaction based diagnosis of mycotic keratitis through single stranded conformation polymorphism. AB - PURPOSE: To report a method for early and correct diagnosis of mycotic keratitis. DESIGN: Clinical laboratory diagnostic study. METHODS: Corneal scraping of all the four patients were processed for DNA extraction which were amplified by fungal specific primers of internal transcribed spacer region I (ITS1). These products were sequenced and analyzed by single stranded conformation polymorphism (SSCP) for species identification. RESULTS: The DNA samples from corneal scrapings of all the four patients were successfully amplified by the primer pair ITS1 and ITS2 and similarity/dissimilarity were established by Jaccard's coefficient. Patient isolate 1 was identified as Nectria hematococca, isolate 2 as Candida albicans, and isolates 3 and 4 were identified as Bipolaris papendorfii. This led to prompt initiation of antifungal therapy in all the four cases where useful vision could be restored. CONCLUSIONS: Early and correct diagnosis of mycotic keratitis by polymerase chain reaction could be obtained in all the four cases compared with conventional methods, which helped in the prompt initiation of antifungal therapy in patients. PMID- 16310464 TI - Assessment of biomechanics of orbital fracture: a study in goats and implications for oculoplastic surgery in humans. AB - PURPOSE: To determine the biophysical characteristics of energy absorption of the various orbital structures and the amount of force/energy required to produce an orbital fracture in goats, and to interpret results in terms of human clinical applicability. DESIGN: Experimental animal study. METHODS: The energy required to fracture the orbit on direct globe impact was measured. We used a variety of other measurements, including high-speed videography, intraocular pressure, and intraorbital pressure measurements to describe the biophysics of floor fracture. Twenty-one goats and 42 orbits were studied in this single institutional site study. RESULTS: Our measurements determined the fracture threshold for the caprine orbit averaged 26.5 N (SE 3.1 N) with a total energy threshold for fracture of 2600 mJ (SE 280 mJ). Eye deformation measured 6 mm with deformation energy dissipation measuring 100 to 180 mJ (SE 10 mJ). Eye displacement measured 10 mm with dissipation measuring 50 to 75 mJ (SE 4.0 mJ). The deconvoluted bone fracture energy averaged 2350 mJ (SE 260 mJ). CONCLUSIONS: Impact energy applied to the orbit through the eyeball is absorbed and dissipated by three primary mechanisms: (1) globe deformation, (2) globe displacement, and (3) orbital fracture. Excess energy (above the energy required for deformation and displacement) >or=2350 mJ will result in an orbital fracture. PMID- 16310465 TI - Decision analysis with Markov processes supports early surgery for large-angle infantile esotropia. AB - PURPOSE: To assess tradeoffs in time spent with aligned visual axes or stereopsis and risked reoperations between three strategies for the treatment of large-angle infantile esotropia: first surgery at 6, 24, or 48 months of age. DESIGN: Decision analysis. METHODS: We simulated three hypothetic cohorts of children with this condition until their eighth year. Potential achievement of stereopsis, the diagnosis of dissociated vertical divergence or inferior oblique overaction, and various complications were modeled. Probability estimates were obtained from a comprehensive literature review. Analyses aimed to determine the strategy that was associated with longer time spent with aligned eyes and/or stereopsis and fewer reoperations. RESULTS: Eight-year-old children are expected to spend 72.6 vs 58.0 vs 37.8 months with aligned eyes and 26.8 vs 9.0 vs 1.5 months with stereopsis and to receive on average 1.76 vs 1.67 vs 1.46 surgeries, if they first underwent operation at 6 vs 24 vs 48 months, respectively. Children at 8 years are expected to have stereopsis at the rate of 36.1% vs 17.2% vs 5.1%, respectively. Operation at 6 months instead of 48 months is expected to yield an additional 9% of total follow-up time in health states with eye alignment and 11% in health states with stereopsis, per 5% increase in risk for extra operations (5% and 7%, respectively, per 5% increase in risk for operation at 6 vs 24 months). Wide-range sensitivity analyses and long-term projections point to the same direction. CONCLUSION: Given the benefit/risk tradeoffs, it is probably warranted to attempt to correct large-angle infantile esotropia as early as possible. PMID- 16310466 TI - The dilemma of hydroxychloroquine screening: new information from the multifocal ERG. PMID- 16310467 TI - Diffusion tensor imaging of the visual sensory pathway: are we there yet? PMID- 16310468 TI - Dry eye (Keratoconjunctivitis Sicca), rheumatoid arthritis, and Sjogren's syndrome. PMID- 16310469 TI - Removing silicone oil from eyes with cytomegalovirus retinitis. PMID- 16310470 TI - Visual training: current status in ophthalmology. AB - PURPOSE: To inform ophthalmologists of the current status of visual training. DESIGN: Personal perspective. METHOD: A perspective and analysis of current practices that include a review of the literature and personal experiences of the author. RESULTS: Visual training of some sort has been used for centuries. In the first half of the twentieth century, in cooperation with ophthalmologists, orthoptists introduced a wide variety of training techniques that were designed primarily to improve binocular function. In the second half of the twentieth century, visual training activities were taken up by optometrists and paramedical personnel to treat conditions that ranged from uncomfortable vision to poor reading or academic performance. Other visual training has been aimed at the elimination of a wide variety of systemic symptoms and for the specific improvement of sight and even for the improvement of athletic performance. At present, ophthalmologists and orthoptists use visual training to a very limited degree. Most visual training is now done by optometrists and others who say it works. Based on an assessment of claims and a study of published data, the consensus of ophthalmologists regarding visual training is that, except for near point of convergence exercises, visual training lacks documented evidence of effectiveness. CONCLUSION: Although visual training has been used for several centuries, it plays a minor and actually decreasing role in eye therapy used by the ophthalmologist. At the beginning of the twenty-first century, most visual training is carried out by non-ophthalmologists and is neither practiced nor endorsed in its broadest sense by ophthalmology. PMID- 16310471 TI - Outcomes of small-incision cataract surgery in eyes with preexisting Ahmed Glaucoma Valves. AB - PURPOSE: To investigate outcomes of phacoemulsification in eyes with functioning Ahmed Glaucoma Valves (AGVs). DESIGN: Observational case series. METHODS: We reviewed records of 19 patients (23 eyes) who had a clear-cornea phacoemulsification >/=3 months after insertion of an AGV between 1997 and 2002. The main outcome measures were intraocular pressure (IOP) control, number of medications, complications, and further glaucoma surgery. RESULTS: The median interval between AGV surgery and phacoemulsification was 0.7 years with an average (+/-SD) follow-up time of 1.6 (+/-0.6) years (range: 0.9 to 3.0 years). Four eyes (17%) had an IOP increase >10 mm Hg on day one. The mean IOP or number of medications did not significantly change after phacoemulsification at one month or thereafter (P > .05). One eye (4%) required a second AGV. CONCLUSIONS: Small-incision cataract surgery is consistent with maintenance of IOP control in eyes with functioning AGV. A minority of the eyes will require repeat glaucoma surgery. PMID- 16310472 TI - Intraocular pressure in Northern China in an urban and rural population: the Beijing eye study. AB - PURPOSE: To evaluate intraocular pressure (IOP) and its demographic associations in an urban and rural population in Northern China. DESIGN: Population-based cohort study. METHODS: In the Beijing Eye Study, IOP was measured by noncontact pneumatonometry. Mean age was 56.20 +/- 10.59 years (range, 40-101 years). RESULTS: Mean IOP was 16.11 +/- 3.39 mm Hg (median, 16 mm Hg; range, 7-60 mm Hg). In the rural group and the urban population group, respectively, IOP was significantly (P = .016 and P = .029, respectively) lower in the high-age group (>or=75 years) than in the middle-age group (55-64 years). In the rural group, IOP was additionally significantly lower in the age group of 40 to 44 years than in the middle-age group, which suggests a reversed U-like course. A multiple regression analysis revealed that IOP significantly increased with myopia (P = .024), and that it was statistically independent of rural area vs urban area (P = .12) and gender (P = .71). CONCLUSION: In the Northern Chinese population, mean and median of IOP that is measured by pneumatonometry is 16 mm Hg with a normal range (mean +/- two SDs) of 10 to 22 mm Hg. In cross-sectional analysis, IOP slightly increased towards an age of 55 to 64 years and decreased towards higher ages. IOP increased with myopia. It was not associated with rural vs urban region or gender. PMID- 16310473 TI - Successful treatment of idiopathic orbital inflammation with mycophenolate mofetil. AB - PURPOSE: To report our experience with the use of mycophenolate mofetil for treatment of idiopathic orbital inflammation (IOI). DESIGN: Noncomparative interventional case series. METHODS: We reviewed the records of five patients with IOI who were treated with mycophenolate mofetil. The indications for use included recurrent inflammation after other therapies (four patients) and to avoid corticosteroid-induced blood sugar elevation in poorly controlled diabetes mellitus (one patient). Treated patients were followed for a median of 8.5 months (range, 4-30 months) after mycophenolate mofetil therapy was begun. Main outcome measures were response to treatment, treatment-related side effects, drug dosage, previous and final treatment regimens, and discontinuation of systemic corticosteroids. RESULTS: Mycophenolate mofetil therapy was discontinued in one patient because of nausea. Each of the remaining four patients experienced complete resolution of orbital inflammation. Of these, only one patient remains on prednisone, but at a significantly reduced dose (1 mg/d). CONCLUSION: Mycophenolate mofetil therapy resulted in the resolution of inflammation in patients with refractory or corticosteroid-dependent IOI and was used successfully for a first episode of IOI in a patient for whom corticosteroids were contraindicated. PMID- 16310474 TI - Long-term safety and efficacy of repositioning dislocated plate haptic intraocular lenses in the ciliary sulcus. AB - PURPOSE: To report safety and efficacy outcomes of repositioning posterior dislocated plate haptic lenses (PHLs) in the ciliary sulcus (CS). DESIGN: Retrospective interventional case series. METHODS: Fifteen consecutive eyes with dislocated PHL repositioned in the CS were reviewed retrospectively. Pars plana vitrectomy was required in 12 of 15 cases. The main outcome variables were initial and final best-corrected visual acuity (BCVA) and length of follow-up. RESULTS: All lenses remained centered for an average of 48.7 months (range 4 to 99 months) of follow-up with no recurrent dislocation, cystoid macular edema, chronic iritis, or iris chaffing. Initial and final BCVA of 20/40 or better was attained in 93.3% (14 of 15) and 66.7% (10 of 15) of eyes, respectively. Decline of final BCVA was attributable to secondary eye diseases and not related to repositioning. CONCLUSIONS: Our study reports dislocated PHL may be managed safely and effectively by placement in the CS with long-term stability under certain clinical scenarios. PMID- 16310475 TI - The effectiveness of isobutyl cyanoacrylate tissue adhesive for the treatment of corneal perforations. AB - PURPOSE: To study the results of treatment of corneal perforations with isobutyl cyanoacrylate tissue adhesive. DESIGN: Retrospective case series. METHODS: The charts of 20 patients (22 eyes) with corneal perforation 125 microm) and/or (2) soft distinct drusen (>125 microm) associated with pigmentary abnormalities. RESULTS: The 3-year incidence of AMD was 0.49% (95% confidence interval [CI]: 0.13 to 0.85) and increased significantly with age, reaching 3.41% (95% CI: 0 to 7.2) in participants aged 80 years or more. After adjustment for age, eyes with early ARM at baseline were 78 times more at risk of developing AMD than eyes without early ARM (OR = 78.4, 95% CI: 14.6 to 420.1). CONCLUSIONS: This study confirms that AMD develops mainly in subjects aged 80 years or older, and in subjects with early ARM. PMID- 16310478 TI - Deep corneal neovascularization after implantation with intrastromal corneal ring segments. AB - PURPOSE: To demonstrate the development of deep corneal neovascularization after intrastromal corneal ring segment (ICRS) implantation, and to demonstrate complete regression after treatment with surgical removal and anti-inflammatory therapy. DESIGN: Observational case report. METHODS: A 29-year-old man developed deep stromal neovascularization after ICRS implantation for post-laser-assisted in situ keratomileusis ectasia that was not associated with the surgical wound. RESULTS: After surgical removal of the ICRS and treatment with topical prednisolone acetate 1.0% and cyclosporine A 1.0%, complete vessel regression occurred in 2 weeks. CONCLUSIONS: ICRS can induce deep corneal neovascularization that is not associated with the surgical wound. Surgical removal of the intrastromal ring and treatment with topical anti-inflammatory agents can induce vessel regression. PMID- 16310479 TI - Association between intensity of posterior capsule opacification and contrast sensitivity. AB - PURPOSE: To investigate the correlation between change in contrast sensitivity (CS) and difference in objective posterior capsule opacification (PCO) score before and after Nd:YAG laser capsulotomy. DESIGN: Prospective interventional case series. METHODS: CS (Functional Acuity Contrast Test, or FACT, chart), distance visual acuity (Early Treatment Diabetic Retinopathy Study [ETDRS] chart), and pupil size of 30 pseudophakic eyes with PCO of varying intensity were measured before and after capsulotomy. Digital retroillumination images were taken, and the area corresponding to the pupil size was evaluated with an automated PCO analysis program known as AQUA. RESULTS: The correlation between change in PCO score and change in CS was highest (r = 0.85) for row B on the FACT chart and lowest (r = 0.09) for row E. The correlation with change in ETDRS visual acuity was 0.57. CONCLUSIONS: Objective PCO assessment by an automated image analysis system correlates well with PCO-induced loss of CS and is therefore an appropriate method for clinical studies on the development and prevention of PCO. PMID- 16310480 TI - Risk of retinopathy in children with type 1 diabetes mellitus before 2 years of age. AB - PURPOSE: To assess the risk of diabetic retinopathy (DR) in children with type 1 diabetes mellitus (T1DM) diagnosed at a very early age. DESIGN: Observational case series. METHODS: The records of 51 patients were identified through the diabetes database of the Division of Pediatric Endocrinology and Metabolism at Washington University School of Medicine. The patients were diagnosed with T1DM before 2 years of age and were monitored for at least 5 years after diagnosis. The results of ophthalmic screening examinations were reviewed. RESULTS: Fifty one patients were identified, 33 of whom were monitored for >8 years. None of the patients developed DR. CONCLUSIONS: Children have a negligible risk of developing DR during the first 10 years of life, even if they are diagnosed before age 2. These results indicate that screening for DR is not necessary before age 10. PMID- 16310481 TI - Methylenetetrahydrofolate reductase genetic polymorphisms in patients with cataract. AB - PURPOSE: Hyperhomocysteinemia is commonly associated with polymorphisms in the methylenetetrahydrofolate reductase (MTHFR) gene. The level of homocysteine can be lowered by dietary intake of folate. A protective effect of folate supplementation has been reported against cataract. Here we investigate MTHFR polymorphisms in human cataract. DESIGN: Retrospective case-control association study. METHODS: Patients with nuclear (n = 77), cortical (n = 155), posterior subcapsular (n = 119), and mixed (n = 151) cataract, and 187 controls were analyzed for the MTHFR 677C-->T and 1298A-->C polymorphisms using minisequencing technique. RESULTS: The wild-type MTHFR 677CC/1298AA genotype was strongly overrepresented among cataract cases (P = .003). This effect was most pronounced in the mixed cataract group (P < .001). Hyperhomocysteinemia-associated genotypes had similar frequencies in cataract and control groups. CONCLUSIONS: The previously reported protective effect of folate against cataract is not due to overrepresentation of hyperhomocysteinemia-associated MTHFR genotypes. Instead, the strong predominance of wild-type MTHFR in cataract may suggest impaired DNA synthesis as a cataractogenic factor. PMID- 16310482 TI - Blepharoptosis and upper eyelid swelling due to lipogranulomatous inflammation caused by silicone oil. AB - PURPOSE: Report of two patients who presented with unilateral upper eyelid swelling and ptosis 1 and 8 years, respectively, after vitreoretinal surgery with intraocular silicone oil. DESIGN: Report of two cases. METHODS: In the first case, the oil was still present in the eye. In the second case, the intraocular oil and an epibulbar buckle had been removed 7 years earlier. RESULTS: Histopathology of upper eyelid skin and preaponeurotic fat revealed lipogranulomatous inflammation. CONCLUSIONS: Leakage of intraocular silicone oil into the eyelid tissues may cause an inflammatory reaction resulting in eyelid swelling and ptosis. We presume that in the first patient, silicone oil had leaked from the eye during or after surgery; in the second patient, silicone oil had probably been left behind in the space previously occupied by the epibulbar buckle. PMID- 16310483 TI - Anaphylactoid reaction after verteporfin therapy. AB - PURPOSE: To report a case of anaphylactoid reaction in a woman after initial exposure to verteporfin and to alert physicians of this potentially life threatening reaction. DESIGN: Interventional case report. METHODS: An 80-year-old woman who was found to have exudative age-related macular degeneration on clinical examination and fluorescein angiography underwent treatment with verteporfin photodynamic therapy (PDT). Thirty minutes after verteporfin PDT infusion, the patient experienced throat constriction, swelling of her hands, and severe shortness of breath. She received immediate intravenous treatment with methylprednisolone, diphenhydramine, and famotidine and was admitted for observation. RESULTS: The patient's symptoms resolved, and she experienced no long-term side effects related to PDT. Routine noninvasive pulse oximetry, however, did result in finger discoloration and superficial burn. Evaluation revealed that her symptoms were noncardiac in origin. CONCLUSION: Verteporfin can result in an anaphylactoid reaction. Treating physicians should be advised of possible photosensitizing reactions once the patient has received verteporfin PDT. PMID- 16310484 TI - Low levels of pigment epithelium-derived factor in highly myopic eyes with chorioretinal atrophy. AB - PURPOSE: To determine the concentration of pigment epithelium-derived factor (PEDF) in the aqueous humor of highly myopic eyes. DESIGN: Observational case series. METHODS: The PEDF concentration in the aqueous humor of 23 eyes of 17 patients with high myopia (axial length >26 mm) who underwent cataract surgery was measured by enzyme-linked immunosorbent assay. RESULTS: The mean concentration of PEDF in eyes with high myopia (0.54 +/- 0.12 microg/ml) was significantly lower than that in control eyes (0.86 +/- 0.04 microg/ml, P = .0022). The PEDF level in myopic eyes with chorioretinal atrophy (0.32 +/- 0.05 microg/ml) was lower than that in myopic eyes without chorioretinal atrophy (0.71 +/- 0.12 microg/ml; P = .041) and control eyes (P = .0003). CONCLUSIONS: The significantly lower concentration of PEDF in eyes with chorioretinal atrophy associated high myopia probably resulted from degeneration of the retinal pigment epithelial cells and/or the retinal ganglion cells that are the main sources of PEDF in the eye. PMID- 16310485 TI - Corneal topographic changes after transconjunctival (25-gauge) sutureless vitrectomy. AB - PURPOSE: To evaluate the topographic changes in the cornea after pars plana vitrectomy (PPV) with 25-gauge transconjunctival sutureless vitrectomy (TSV) system. DESIGN: Prospective, interventional case series. METHODS: In this prospective study, we evaluated the topographic changes of the cornea in 32 eyes of 32 patients who underwent PPV with the 25-gauge TSV system. The topographic parameters that were analyzed statistically were the average corneal power, corneal surface cylinder, surface asymmetry index, and surface regularity index. Mean induced astigmatism was estimated by vector analysis software. Wilcoxon test was used for statistical analysis. RESULTS: There was no significant change in average corneal power, corneal surface cylinder, surface asymmetry index, and surface regularity index parameters at first day, first week, and first month after the operation. Mean induced astigmatism was 0.38 diopters at 15 degrees. CONCLUSIONS: Corneal surface and astigmatic changes were observed to be insignificant in the early postoperative period after PPV with the 25-gauge TSV system. PMID- 16310486 TI - Explantation of an AcrySof Natural intraocular lens because of a color vision disturbance. AB - PURPOSE: To describe a patient who complained of color vision disturbance after implantation of a clear intraocular lens (IOL) in one eye and a yellow IOL in the other. DESIGN: Interventional case report. METHODS: A 50-year-old woman complained of binocular imbalance and "yellow vision" in her left eye after implantation of a visible blue light-filtering yellow IOL. A clear IOL had been implanted in her right eye 1 month before. She requested explantation of the yellow IOL. RESULTS: The yellow IOL was exchanged for a clear one, which resolved the unbalanced color vision. CONCLUSIONS: Patients may be intolerant of the color disturbance that results when dissimilarly colored IOLs are implanted. It is advisable to avoid this practice. PMID- 16310487 TI - Bilateral diffuse uveal melanocytic proliferation in a patient with cancer associated retinopathy. AB - PURPOSE: To describe a patient with bilateral diffuse uveal melanocytic proliferation (BDUMP) and cancer-associated retinopathy (CAR). DESIGN: Interventional case report. METHODS: A 66-year-old woman developed progressive vision loss 4 months after total hysterectomy. Ophthalmologic examination, Western blot test of sera and aqueous humor, and immunohistochemistry of carcinoma cells were performed. RESULTS: Testing revealed BDUMP and severe retinal dysfunction. Autoantibodies against recoverin and heat shock cognate protein 70 (hsc 70) were detected in serum. Cytoplasmic immunoreactivity for recoverin and hsc 70 was observed in endometrioid carcinoma cells. CONCLUSIONS: Simultaneous cases of BDUMP and CAR are rare. Aberrantly expressed recoverin and hsc 70 triggered serum autoantibody production, which caused photoreceptor degeneration. PMID- 16310488 TI - Diagnosis and anatomic mapping of an orbital varix by computed tomographic angiography. AB - PURPOSE: To report the use of computed tomographic angiography (CTA) to diagnose an orbital varix. DESIGN: Observational case report. METHODS: A 73-year-old female was investigated by CTA for carotid artery stenosis. An incidental orbital lesion was discovered. Reformations were obtained. RESULTS: CTA demonstrated a homogeneously enhancing orbital apex lesion. It provided exceptional anatomic detail of inflow and outflow veins, confirming the diagnosis of an orbital varix. CONCLUSIONS: CTA is cost effective, fast, non-invasive, and readily available. These aspects combined with the exceptional vascular detail obtained may make CTA the primary test for orbital varix diagnosis. PMID- 16310489 TI - The effect of subtenon triamcinolone acetonide injection for diabetic macular edema on retinal and choroidal circulation. AB - PURPOSE: To evaluate changes in retinal and choroidal circulation after subtenon triamcinolone acetonide (TA) injection for diabetic macular edema. DESIGN: Prospective interventional case series. METHODS: Thirteen eyes of 13 patients with diabetic macular edema were studied. Fluorescein and indocyanine green angiograms were performed at three periods: before the injection and 1 week and 6 months after subtenon injection of TA (40 mg). Retinal arteriovenous passage time (as an indicator of retinal circulation) and choroidal tau (as an indicator of early filling velocity of choroid) were obtained with image analysis software. RESULTS: Choroidal tau values before and 1 week after subtenon TA injection were, respectively, 3.2 +/- 0.4 and 4.0 +/- 0.7 seconds, which showed a significant delay (P = .01, Wilcoxon signed-rank test). The delayed choroidal tau values returned to pretreatment level at 6 months after TA injection. In contrast, the arteriovenous passage time remained unchanged. CONCLUSION: Subtenon TA injection transiently influences choroidal blood flow. PMID- 16310490 TI - Vogt-Koyanagi-Harada disease in patients with chronic hepatitis C. AB - PURPOSE: To report the cases of four patients with hepatitis C virus infection who experienced clinical features that are virtually identical to Vogt-Koyanagi Harada disease (VKH). DESIGN: Retrospective observational case series. METHODS: Medical records of patients who were referred between January and December 2003 were reviewed for diagnosis and management of VKH and who also had chronic hepatitis C virus (HCV) infection. RESULTS: Four white patients had the clinical features of VKH. Three of the patients experienced intraocular inflammation while they were being treated for HCV infection with pegylated interferon alpha 2b and ribavirin. The intraocular inflammation responded to systemic corticosteroid treatment and to discontinuation of antiviral agents. CONCLUSION: Although the number of patients who were studied is limited, there appears to be an association between HCV infection that was treated with pegylated interferon alpha 2b and the development of VKH-like disease. Further studies are required to confirm such an association. PMID- 16310491 TI - Management of severe hypotony with intravitreal injection of viscoelastic. AB - PURPOSE: To investigate the effect of an intravitreal injection of sodium hyaluronate 1.4% and 2.3% in the treatment of hypotony. DESIGN: Interventional case series. METHODS: Intravitreal injection of sodium hyaluronate 1.4% was performed to treat persistent hypotony in three eyes after the successful repair of retinal detachment and in one eye with uveitis. Injection of sodium hyaluronate 2.3% was performed in one eye after the successful repair of retinal detachment with removal of silicone oil. RESULTS: At the last follow-up examination (2-16 months), intraocular pressure increased to at least 5 mm Hg (5 14 mm Hg) in all eyes that had been injected with sodium hyaluronate. Intraocular pressure increased to 38 mm Hg in the eye that had been injected with sodium hyaluronate 2.3% on postoperative day 1 and was 10 mm Hg at three months. Vision improved in all patients. CONCLUSION: Intraocular injection of sodium hyaluronate 1.4 and 2.3% appears to be efficacious in reversing hypotony in some eyes. Functional and structural findings can also improve. PMID- 16310493 TI - Influence of wavefront aberration and corneal subepithelial haze on low-contrast visual acuity after photorefractive keratectomy. PMID- 16310494 TI - Ocular higher-order aberrations in eyes with supernormal vision. PMID- 16310496 TI - Ocular higher-order aberrations in eyes with supernormal vision. PMID- 16310498 TI - Posterior subtenon triamcinolone acetonide for refractory diabetic macular edema. PMID- 16310499 TI - Optic disk size correlated with refractive error. PMID- 16310500 TI - Persistent diabetic macular edema is associated with elevated hemoglobin A1c. PMID- 16310504 TI - Retinal cystoid spaces in acute Vogt-Koyanagi-Harada syndrome. PMID- 16310506 TI - Pyogenic granulomas after silicone punctal plugs: a clinical and histopathologic study. PMID- 16310507 TI - Modified grid laser photocoagulation versus pars plana vitrectomy with internal limiting membrane removal in diabetic macular edema. PMID- 16310509 TI - Phosphatidylserine decarboxylases as genetic and biochemical tools for studying phospholipid traffic. PMID- 16310510 TI - Blunted activation in orbitofrontal cortex during mania: a functional magnetic resonance imaging study. AB - BACKGROUND: Patients with bipolar disorder have been reported to have abnormal cortical function during mania. In this study, we sought to investigate neural activity in the frontal lobe during mania, using functional magnetic resonance imaging (fMRI). Specifically, we sought to evaluate activation in the lateral orbitofrontal cortex, a brain region that is normally activated during activities that require response inhibition. METHODS: Eleven manic subjects and 13 control subjects underwent fMRI while performing the Go-NoGo task, a neuropsychological paradigm known to activate the orbitofrontal cortex in normal subjects. Patterns of whole-brain activation during fMRI scanning were determined with statistical parametric mapping. Contrasts were made for each subject for the NoGo minus Go conditions. Contrasts were used in a second-level analysis with subject as a random factor. RESULTS: Functional MRI data revealed robust activation of the right orbitofrontal cortex (Brodmann's area [BA] 47) in control subjects but not in manic subjects. Random-effects analyses demonstrated significantly less magnitude in signal intensity in the right lateral orbitofrontal cortex (BA 47), right hippocampus, and left cingulate (BA 24) in manic compared with control subjects. CONCLUSIONS: Mania is associated with a significant attenuation of task related activation of right lateral orbitofrontal function. This lack of activation of a brain region that is usually involved in suppression of responses might account for some of the disinhibition seen in mania. In addition, hippocampal and cingulate activation seem to be decreased. The relationship between this reduced function and the symptoms of mania remain to be further explored. PMID- 16310512 TI - From the molecular genetics revolution to gene therapy: translating basic research into medicine. PMID- 16310513 TI - Influence of hypoalbuminemia or hyperalbuminemia on the serum anion gap. AB - BACKGROUND: Conflicting data exist as to what extent hypoalbuminemia reduces the anion gap; estimates range from 1.5 to 2.5 mM per g/dL decrease in serum albumin. METHODS: We measured serum albumin, total protein, and electrolyte concentrations in 5328 consecutive patients aged 1 month to 102 years. Most patients (3750; 70%) had a normal albumin, but 1158 had hypoalbuminemia (< or =3.4 g/dL); 420 had hyperalbuminemia (> or =4.7 g/dL). Relationships between serum albumin or total protein and the anion gap were analyzed by linear regression. RESULTS: 309 (27%) hypoalbuminemic patients had a decreased anion gap, and 257 hyperalbuminemic patients (61%) had an increased anion gap. Among the entire group of 5328 patients, there were highly significant correlations between either serum albumin or total protein and the anion gap (P < 0.001). The slope of the regression for albumin versus anion gap was 2.3 mM per g/dL. Using this slope, anion gap could be adjusted for abnormal serum albumin levels: anion gap(adjusted) =anion gap + 2.3 (4-albumin). The initial assessment of an anion gap as being increased, normal, or decreased changed in 44% of the patients with hypo- or hyperalbuminemia once anion gap had been adjusted with this formula. CONCLUSIONS: Before considering whether a disorder associated with an increased or decreased anion gap is present, the anion gap should be first adjusted for abnormal serum albumin concentrations. Our data suggest that physicians use 2.3 times the change in serum albumin, whereas those of Figge et al suggest 2.5; either approach gives similar results. PMID- 16310514 TI - Detection of RhD(el) in RhD-negative persons in clinical laboratory. AB - The Rhesus (Rh) blood group is the most polymorphic human blood group system, and it is clinically significant in transfusion medicine. About 15% of Caucasoid people are RhD-negative, whereas in the Asian population, the RhD-negative blood type only occurs in 0.1% to 0.5%. However, approximately 30% of apparently RhD negative Taiwanese people actually were RhD(el). Traditionally, we verify RhD(el) by a serologically adsorption-elution procedure with polyclonal anti-D. In our recent report, RhC phenotype is highly associated with RhD(el), and RHD1227A is a useful genetic marker for RhD(el). For setting up a rapid protocol to detect RhD(el) in clinical laboratory, a total number of 395 Taiwanese serological RhD negative blood samples, those with RhC (+) phenotypes as selected by serological tests, were further screened by adsorption/elution tests and RHD1227A allele by specific sequence primer-polymerase chain reaction (SSP-PCR) for RhD(el). Among 395 blood samples collected from RhD-negative subjects, the incidence of RhC (+) was 43% (171/395). One hundred and twenty six of the 171 RhC (+) samples were positive for both adsorption/elution for RhD detection and SSP-PCR assay for RHD1227A. The sensitivity and specificity were 96.9% and 97.5%, respectively, for RHD1227A detection as compared with the traditional adsorption/elution test. Our results also indicated that RHD1227A was highly linked to Ce haplotypes (95.2%). In conclusion, combined RhC (+) phenotyping and RHD1227A analysis can be a simple and accurate laboratory screening protocol for RhD(el) detection in RhD-negative population. PMID- 16310515 TI - Differential regulation of the rat heme oxygenase-1 expression by Ets oncoproteins in glomerular mesangial cells. AB - The Ets-1 oncoprotein and the heme-catabolizing enzyme heme oxygenase (HO)-1 have been implicated in the pathogenesis of renal disease. We investigated the role of the putative Ets-binding sites (EBSs) in the transactivation of the proximal promoter of rat heme oxygenase 1 (hmox1) gene by the Ets oncoproteins Fli-1, Erg 2, and Ets-1 in mesangial cells. We examined several rat hmox1-chloramphenicol acetytransferase (CAT) constructs and EBS mutant constructs in an effort to assess the effect of ETS oncoproteins on transactivation of the rat hmox1 proximal promoter in renal glomerular mesangial cells. CAT assays demonstrated that the proximal promoter region (-1387 to -40) contains positive and negative regulatory regions and that the EBS-2, 3, and 4 play a role in basal promoter activity. Overexpression of Fli-1 and Erg-2 proteins showed a significant increase in promoter activity, whereas Ets-1 showed no effect on promoter activity. The Fli-1-induced transcriptional activation was not altered by mutation of EBSs, either independently or in combination. However, mutation of EBS-4 independently or a combined mutation of sites 3 and 4 led to a 50% reduction in Erg-2-induced transcriptional activation. Furthermore, mutation of EBS-2 and 4 completely abolished Erg-2-mediated promoter activation. Our results support a role for Ets transcription factors in the regulation of rat hmox-1 gene expression in mesangial cells. PMID- 16310516 TI - Stability of blood cell counts, hematologic parameters and reticulocytes indexes on the Advia A120 hematologic analyzer. AB - Delayed sample analysis is not a rare circumstance in clinical and laboratory practice, especially when blood samples are shipped to distant centralized laboratories, when the analysis can not be readily performed, or when retesting is appropriate. In this study we sought to evaluate the stability of conventional and new hematologic parameters in blood specimens stored for as long as 24 hours at 4 degrees C. Of the 21 hematologic parameters tested with the use of the Advia 120 hematologic analyzer (Bayer Diagnostics), means for paired samples of specimens differed significantly over the 24-hour storage period for hematocrit, main corpuscular volume, percentage of macrocytes, platelet count, main platelet volume, reticulocyte count and percentage, and reticulocyte hemoglobin content (all P < .01). We noted no significant changes in the other parameters tested or in the white blood cell differential. The overall distribution of the immature reticulocytes fractions remained substantially unchanged, though the high staining-intensity fraction showed a considerable shift from the baseline measure. Bland-Altman plots and limits-of-agreement analysis showed mean biases between -4.8% and 37.2% and relative coefficients of variations ranging from 0.4% to 32.7%. The 95% agreement interval in the set of differences was satisfactory and almost within the current analytic-quality specifications for desirable bias. The results of this investigation suggest that, within certain limitations for parameters derived or calculated from cellular volumes, blood specimens stored for as long as 24 hours at 4 degrees C may be suitable for hematologic testing. PMID- 16310517 TI - Laboratory testing for the antibodies that cause heparin-induced thrombocytopenia: how much class do we need? AB - Heparin-induced thrombocytopenia (HIT) is usually caused by platelet-activating antibodies of immunoglobulin G class that recognize platelet factor-4 (PF4) bound to heparin or certain other polyanions. Commercial enzyme immunoassays (EIAs) for PF4/polyanion-reactive antibodies detect two immunoglobulin classes (IgA and IgM) besides IgG. To investigate whether the additional detection of these antibody classes improves or worsens assay operating characteristics, we compared the sensitivity and specificity of EIAs that detect these 3 immunoglobulin classes individually with that of a commercial EIA (Genetic Testing Institute, GTI), as well as a platelet-activation assay, the serotonin-release assay (SRA). We compared the operating characteristics of these 5 assays by evaluating 448 patients, in 14 of whom clinical HIT developed, who received either unfractionated or low molecular weight heparin in prospective studies that included systematic platelet-count monitoring and serologic evaluation for anti PF4/polyanion antibodies. We found that the SRA and IgG and commercial EIAs had similar high sensitivity for HIT; however, diagnostic specificity (for unfractionated and low molecular weight heparin, respectively) varied considerably, as follows: SRA (95.1%, 97.2%) > IgG EIA (89.0%, 93.7%) > GTI EIA (74.2%, 87.6%). Additional detection of IgA and IgM antibodies by the GTI EIA worsened test specificity by detecting numerous nonpathogenic antibodies. Moreover, the frequency and magnitude of IgA and IgM antibody formation in non HIT immune responses did not differ from that exhibited by patients in whom clinical HIT developed. We conclude that an EIA that detects anti-PF4/polyanion antibodies of only the IgG class has greater diagnostic usefulness in revealing clinical HIT than does an assay that also detects IgA and IgM class antibodies. PMID- 16310518 TI - Serum levels of C-reactive protein and procalcitonin in critically ill patients with cirrhosis of the liver. AB - Concentrations of C-reactive protein (CRP) and procalcitonin (PCT) have been suggested as markers of infection. The liver is believed to be a key source of CRP and PCT. For this reason we assessed the predictive value of these markers in patients with hepatic cirrhosis in a 31-bed university-hospital department of intensive care. Demographic, clinical, laboratory, and microbiologic data were collected prospectively over 9 months. Of 864 patients included in the study, 79 (9%) had hepatic cirrhosis. Patients with cirrhosis were more likely to have a medical than a surgical admission diagnosis (67 vs 47%, P = .03). They also had a higher rate of infection (48 vs 30%, P = .03) and higher mortality (44 vs 17%, P = .01) than did patients without cirrhosis. We detected no differences in CRP and PCT concentrations among patients with cirrhosis and different disease severity as assessed on the basis of Child-Pugh score. The serum CRP concentration (admission 11.2 +/- 4.6 vs 13.0 +/- 5.8, maximum 13.9 +/- 6.4 vs 18.8 +/- 7.3 mg/dL) and PCT (admission 1.3 +/- 0.9 vs 2.0 +/- 1.4, maximum 3.3 +/- 1.8 vs 3.4 +/- 2.1 ng/mL) were slightly lower in infected patients with cirrhosis than in infected patients without cirrhosis, but the differences were not statistically significant. Although the liver is considered the main source of CRP and a source of PCT, serum levels of these acute-phase proteins are not significantly lower in patients with cirrhosis than in other patients. Moreover, the predictive power of CRP and PCT for infection was similar for patients with and without cirrhosis. PMID- 16310519 TI - The quack doctor. PMID- 16310521 TI - Genetics of allergic disease. AB - Currently, more than 20.3 million Americans report having asthma and an even greater number suffer from allergies. The cost for treatment of these dis-eases in the United States is greater than $8 billion with more than 40% of this total representing drug expenditure [59]. An intense effort has been made to understand the genetic components of asthma and allergies and how the identified genetic differences influence disease progression and response to drugs. In the future, it will be possible in the clinical setting to analyze a patient's genetic repertoire. From this information, the physician will gain in-sight into the genes involved in producing that subject's allergic and asthmatic phenotype; understand the natural history of that patient's disease;and predict responses (positive and negative) to pharmacologic agents. The end result will be the ability to tailor a specific treatment regime for each patient and reduce the overall cost of health care related to allergies and asthma. PMID- 16310522 TI - Allergic rhinitis: impact of the disease and considerations for management. AB - AR is a common condition affecting individuals of all ages. Those afflicted with AR often suffer from associated inflammatory conditions of the mucosa,such as AC, rhinosinusitis, asthma, otitis media with effusion, and other atopic conditions, such as eczema and food allergies. Lack of treatment or treatment with suboptimal therapy may result in reduced quality of life and compromise productivity at work or school. Although environmental controls may prove difficult to implement, and not all controls appear adequately to mitigate symptoms of AR, they continue to represent a foundation for treatment. Many different classes of medications are now available, and they have been shown to be effective and safe in a large number of well-designed, double-blind, placebo-controlled clinical trials. Some of the over-the-counter medi-cations have been associated with increased sedation, potentially leading to accidents and fatalities at work or while operating complex machinery, such as automobiles. Only immunotherapy with increasing doses of individually targeted allergens results in sustained changes in the immune system. Although anti-IgE is probably only the first successful immunomodulator commercially available to treat AR, monoclonal antibodies will remain too costly, at least in the near future, to find their way into routine AR treatment. PMID- 16310523 TI - Asthma: diagnosis and management. AB - The diagnosis and management of asthma continue to be of critical importance, as recent trends have demonstrated its increasing prevalence, morbidity, and perhaps mortality. Because current treatments for asthma are effective and safe, it is important to diagnose asthma early and to use treatments effectively, particularly those directed toward airway inflammation. The diagnostic measures and array of medications, both those currently available and on the horizon, provide an armamentarium for effective diagnosis, management, and monitoring of asthma. In the coming years, it is expected that additional testing modalities will be available for more precise monitoring of asthma control, and an increased understanding of pharmacogenetics will enable the tailoring of asthma medications to specific patients, providing customized therapy to maximize asthma control. PMID- 16310524 TI - Differential diagnosis of adult asthma. AB - Although the term "all that wheezes is not asthma" is not new, and the long list of asthma masqueraders has remained essentially the same for several decades, the importance of knowing when to question the accuracy ofa diagnosis of asthma has remained critical for physicians who care for patients with respiratory symptoms. The concepts of "asthma control" and"asthma severity" are currently evolving, although the fundamental hall-marks that define the syndrome of asthma endure and should be mastered by asthma specialists. Asthma masqueraders, including several that may confound a correct diagnosis of asthma, are important to consider when either the presentation of asthma is atypical or the response of the patient to treatment is suboptimal. COPD and VCD head the list of diagnoses most likely to be confused with asthma in everyday practice. Correctly identifying the diagnosis of COPD enables implementation of an up-to-date treatment plan that differs from asthma management. VCD is a vastly under recognized syndrome whose existence is widely accepted but whose pathophysiology is poorly understood, and correctly identifying a VCD component to asthma symptoms enables both a reduction in costly and potentially harmful asthma medications and focus on specific VCD treatment, such as speech therapy. For less common and uncommon asthma masqueraders, it is important to be familiar with their typical clinical presentation and basic diagnostic approaches. PMID- 16310525 TI - Anaphylaxis. AB - Anaphylaxis is an acute multisystem allergic reaction that is potentially fatal. Anaphylactic episodes are most commonly caused by foods or drugs, but in many instances have no known cause. Each physician should be equipped in office for therapy of the acute event. The drug of choice, which should be administered immediately, is epinephrine. Although there is some debate as to the preferred injection site, it is clear that of sites studied to date, injection in the lateral thigh (vastus lateralis) produces the most rapid rise in serum level. Any patient predisposed to anaphylactic episodes should wear identifying medical jewelry and avoid, whenever possible, drugs that could worsen an event or complicate its therapy. PMID- 16310526 TI - Adverse reactions to foods. AB - Food allergy encompasses a variety of immune-mediated adverse reactions to foods. IgE-mediated, cell-mediated, and mixed-mechanism food allergy disorders are recognized. Over the past 2 decades, the prevalence of food allergy doubled and its phenotypic expression increased in Westernized societies. Major food allergens have been identified for many common foods. Laboratory diagnosis of food allergy relies heavily on the detection of food-specific IgE antibodies, but novel approaches include tests for T-cell-mediated disorders and tests for prediction of tolerance. OFC remains the diagnostic standard for food allergy. Management of food allergy focuses on avoidance of the offending foods, nutritional support, and prompt recognition and treatment of acute food allergic reactions. Anti-IgE monoclonal antibody is the first potential therapy for food allergy that is under-going testing in clinical trials. PMID- 16310527 TI - Allergic diseases of the eye. AB - The prevalence of ocular allergy is clearly underappreciated; it has been an underdiagnosed and undertreated area in primary care medicine. The ocular symptoms associated with the most common ocular allergy conditions,such as seasonal and perennial AC, are twice as likely to affect the allergy sufferer as nasal symptoms alone. The emergence of new medications for the specific treatment of ocular symptoms over the course of the past 15 years offers a new field for improved patient care by the primary and sub-specialty health care providers. PMID- 16310528 TI - Atopic dermatitis. AB - Atopic dermatitis (AD) is an eczematous, highly pruritic chronic inflammatory skin disease. It usually begins early in life and often occurs in people with a personal or family history of asthma and allergic rhinitis. The prevalence is high, especially in children,and it has been rising in recent decades, in parallel with asthma prevalence. Although AD is often described as an "allergic" dis-ease, allergic causation is difficult to document, and AD is increasingly viewed as a skin disease that predisposes to allergies. This interpretation, based on clinical, epidemiologic, and animal stu-dies, may greatly influence our approach to therapy and prevention of atopic diseases in the coming years. PMID- 16310529 TI - Allergic contact dermatitis. AB - The pathophysiology of ACD follows an intricate design and results in the characteristic, delayed inflammatory response. Although the astute physician may correctly diagnose ACD from its initial, classic history and presentation, alternative diagnoses should be considered and excluded. Patch testing performed with a relevant panel of contact allergens is the ultimate confirmatory test of ACD. Correctly identifying the inciting allergen permits appropriate personal avoidance. Corticosteroids remain the principal treatment options. PMID- 16310530 TI - Urticaria: selected highlights and recent advances. AB - Urticaria has been called a vexing problem and remains so today. The most important part of the diagnostic evaluation remains a comprehensive and detailed history and physical examination, supplemented with limited laboratory testing. Although acute urticaria has been relatively well understood for some time, significant and important recent advances in under-standing the pathogenesis of chronic urticaria are beginning to provide insight in this challenging field, notably the identification of many of these patients with an autoimmune etiology. Antihistamines of various types continue to represent the keystone of symptomatic treatment, with adjunctive support from medications of other classes, such as antileukotrienes, adrenergics, and immunosuppressive and anti-inflammatory agents (including steroids and cyclosporine). Although some progress has been made at improving symptomatic control of urticaria, further research and discovery are necessary before there can yet be an effective impact on the underlying course and natural history of this condition. PMID- 16310531 TI - Insect sting allergy. AB - Insect sting allergy has served as an excellent model for the allergic process over the past century. In particular, during the last 30 years, a new form of diagnostic testing and treatment with vo m has been one of the great suc-cess stories in the entire field of allergy. VIT reduces the risk of recurrent life threatening reactions from about 60% to less than 2%. Progress and further questions continue with a search for a definitive diagnostic test that more accurately predicts which patients are at risk for future reactions, and defines which patients can stop VIT and which ones need to continue treatment. PMID- 16310532 TI - Drug hypersensitivity. AB - Allergic drug reactions compose a small percentage of ADRs, yet they are commonly encountered in clinical practice, and physicians are taught routinely to question patients about these reactions during history taking. Among antibiotics, the immunochemistry of penicillins has been elucidated,leading to the development of validated skin test reagents to diagnose type 1 allergy. Currently, the temporary commercial unavailability of Pre-Pen makes accurate penicillin skin testing impossible; however, this important skin test reagent is expected to become available sometime in 2006. Type 1 allergies to most other drugs lack comparable diagnostic tests, and their diagnosis is therefore driven by the patient's history. When readministration of medications to which patients report previous reactions is indicated, it may be almost always successfully accomplished by means of either graded challenge or desensitization. PMID- 16310533 TI - Politics trumps science at the FDA. PMID- 16310535 TI - The UK drug industry: responsible, ethical, and professional? PMID- 16310534 TI - Maintaining anti-AIDS commitment post "3 by 5". PMID- 16310536 TI - After FIELD: should fibrates be used to prevent cardiovascular disease in diabetes? PMID- 16310537 TI - Total lymphocyte counts and ART in resource-limited settings. PMID- 16310538 TI - COPD: good lung health is the key. PMID- 16310539 TI - HIV risks in incarcerated injection-drug users. PMID- 16310540 TI - Seeking rational policy settings for PMTCT. PMID- 16310541 TI - Ashok Alexander: taking on the challenge of AIDS in India. PMID- 16310542 TI - Roflumilast for chronic obstructive pulmonary disease. PMID- 16310543 TI - Roflumilast for chronic obstructive pulmonary disease. PMID- 16310544 TI - Roflumilast for chronic obstructive pulmonary disease. PMID- 16310545 TI - Roflumilast for chronic obstructive pulmonary disease. PMID- 16310547 TI - Roflumilast for chronic obstructive pulmonary disease. PMID- 16310548 TI - Adherence to dietary advice in weight-reduction drug trials. PMID- 16310549 TI - Incivility in hospitals: health workers strike back. PMID- 16310550 TI - End-of-life issues. PMID- 16310551 TI - Effects of long-term fenofibrate therapy on cardiovascular events in 9795 people with type 2 diabetes mellitus (the FIELD study): randomised controlled trial. AB - BACKGROUND: Patients with type 2 diabetes mellitus are at increased risk of cardiovascular disease, partly owing to dyslipidaemia, which can be amenable to fibrate therapy. We designed the Fenofibrate Intervention and Event Lowering in Diabetes (FIELD) study to assess the effect of fenofibrate on cardiovascular disease events in these patients. METHODS: We did a multinational, randomised controlled trial with 9795 participants aged 50-75 years, with type 2 diabetes mellitus, and not taking statin therapy at study entry. After a placebo and a fenofibrate run-in phase, we randomly assigned patients (2131 with previous cardiovascular disease and 7664 without) with a total-cholesterol concentration of 3.0-6.5 mmol/L and a total-cholesterol/HDL-cholesterol ratio of 4.0 or more or plasma triglyceride of 1.0-5.0 mmol/L to micronised fenofibrate 200 mg daily (n=4895) or matching placebo (n=4900). Our primary outcome was coronary events (coronary heart disease death or non-fatal myocardial infarction); the outcome for prespecified subgroup analyses was total cardiovascular events (the composite of cardiovascular death, myocardial infarction, stroke, and coronary and carotid revascularisation). Analysis was by intention to treat. The study was prospectively registered (number ISRCTN 64783481). FINDINGS: Vital status was confirmed on all but 22 patients. Averaged over the 5 years' study duration, similar proportions in each group discontinued study medication (10% placebo vs 11% fenofibrate) and more patients allocated placebo (17%) than fenofibrate (8%; p<0.0001) commenced other lipid treatments, predominantly statins. 5.9% (n=288) of patients on placebo and 5.2% (n=256) of those on fenofibrate had a coronary event (relative reduction of 11%; hazard ratio [HR] 0.89, 95% CI 0.75-1.05; p=0.16). This finding corresponds to a significant 24% reduction in non-fatal myocardial infarction (0.76, 0.62-0.94; p=0.010) and a non-significant increase in coronary heart disease mortality (1.19, 0.90-1.57; p=0.22). Total cardiovascular disease events were significantly reduced from 13.9% to 12.5% (0.89, 0.80-0.99; p=0.035). This finding included a 21% reduction in coronary revascularisation (0.79, 0.68-0.93; p=0.003). Total mortality was 6.6% in the placebo group and 7.3% in the fenofibrate group (p=0.18). Fenofibrate was associated with less albuminuria progression (p=0.002), and less retinopathy needing laser treatment (5.2%vs 3.6%, p=0.0003). There was a slight increase in pancreatitis (0.5%vs 0.8%, p=0.031) and pulmonary embolism (0.7%vs 1.1%, p=0.022), but no other significant adverse effects. INTERPRETATION: Fenofibrate did not significantly reduce the risk of the primary outcome of coronary events. It did reduce total cardiovascular events, mainly due to fewer non-fatal myocardial infarctions and revascularisations. The higher rate of starting statin therapy in patients allocated placebo might have masked a moderately larger treatment benefit. PMID- 16310552 TI - Safety and efficacy of zinc supplementation for children with HIV-1 infection in South Africa: a randomised double-blind placebo-controlled trial. AB - BACKGROUND: Zinc deficiency is associated with impaired immune function and an increased risk of infection. Supplementation can decrease the incidence of diarrhoea and pneumonia in children in resource-poor countries. However, in children with HIV-1 infection, the safety of zinc supplementation is uncertain. We aimed to assess the role of zinc in HIV-1 replication before mass zinc supplementation is recommended in regions of high HIV-1 prevalence. METHODS: We did a randomised double-blind placebo-controlled equivalence trial of zinc supplementation at Grey's Hospital in Pietermaritzburg, South Africa. 96 children with HIV-1 infection were randomly assigned to receive 10 mg of elemental zinc as sulphate or placebo daily for 6 months. Baseline measurements of plasma HIV-1 viral load and the percentage of CD4+ T lymphocytes were established at two study visits before randomisation, and measurements were repeated 3, 6, and 9 months after the start of supplementation. The primary outcome measure was plasma HIV-1 viral load. Analysis was per protocol. FINDINGS: The mean log(10) HIV-1 viral load was 5.4 (SD 0.61) for the placebo group and 5.4 (SD 0.66) for the zinc supplemented group 6 months after supplementation began (difference 0.0002, 95% CI -0.27 to 0.27). 3 months after supplementation ended, the corresponding values were 5.5 (SD 0.77) and 5.4 (SD 0.61), a difference of 0.05 (-0.24 to 0.35). The mean percentage of CD4+ T lymphocytes and median haemoglobin concentrations were also similar between the two groups after zinc supplementation. Two deaths occurred in the zinc supplementation group and seven in the placebo group (p=0.1). Children given zinc supplementation were less likely to get watery diarrhoea than those given placebo. Watery diarrhoea was diagnosed at 30 (7.4%) of 407 clinic visits in the zinc-supplemented group versus 65 (14.5%) of 447 visits in the placebo group (p=0.001). INTERPRETATION: Zinc supplementation of HIV-1-infected children does not result in an increase in plasma HIV-1 viral load and could reduce morbidity caused by diarrhoea. RELEVANCE TO PRACTICE: Programmes to enhance zinc intake in deficient populations with a high prevalence of HIV-1 infection can be implemented without concern for adverse effects on HIV-1 replication. In view of the reductions in diarrhoea and pneumonia morbidity, zinc supplementation should be used as adjunct therapy for children with HIV-1 infection. PMID- 16310553 TI - Use of total lymphocyte count for informing when to start antiretroviral therapy in HIV-infected children: a meta-analysis of longitudinal data. AB - BACKGROUND: Total lymphocyte count has been proposed as an alternative to the percentage of CD4+ T-cells to indicate when antiretroviral therapy should be started in children with HIV in resource-poor settings. We aimed to assess thresholds of total lymphocyte count at which antiretroviral therapy should be considered, and compared monitoring of total lymphocyte count with monitoring of CD4-cell percentage. METHODS: Longitudinal data on 3917 children with HIV infection were pooled from observational and randomised studies in Europe and the USA. The 12-month risks of death and AIDS by most recent total lymphocyte count and age were estimated by parametric survival models, based on measurements before antiretroviral therapy or during zidovudine monotherapy. Risks were derived and compared at thresholds of total lymphocyte count and CD4-cell percentage for starting antiretroviral therapy recommended in WHO 2003 guidelines. FINDINGS: Total lymphocyte count was a powerful predictor of the risk of disease progression despite a weak correlation with CD4-cell percentage (r=0.08-0.19 dependent on age). For children older than 2 years, the 12-month risk of death and AIDS increased sharply at values less than 1500-2000 cells per muL, with little trend at higher values. Younger children had higher risks and total lymphocyte count was less prognostic. Mortality risk was substantially higher at thresholds of total lymphocyte count recommended by WHO than at corresponding thresholds of CD4-cell percentage. When the markers were compared at the threshold values at which mortality risks were about equal, total lymphocyte count was as effective as CD4-cell percentage for identifying children before death, but resulted in an earlier start of antiretroviral therapy. INTERPRETATION: In this population, total lymphocyte count was a strong predictor of short-term disease progression, being only marginally less predictive than CD4 cell percentage. Confirmatory studies in resource-poor settings are needed to identify the most cost-effective markers to guide initiation of antiretroviral therapy. PMID- 16310554 TI - Chronic obstructive pulmonary disease in five Latin American cities (the PLATINO study): a prevalence study. AB - BACKGROUND: Both the prevalence and mortality attributable to chronic obstructive pulmonary disease (COPD) seem to be increasing in low-income and middle-income countries, but few data are available. The aim of the PLATINO study, launched in 2002, was to describe the epidemiology of COPD in five major Latin American cities: Sao Paulo (Brazil), Santiago (Chile), Mexico City (Mexico), Montevideo (Uruguay), and Caracas (Venezuela). METHODS: A two-stage sampling strategy was used in the five areas to obtain probability samples of adults aged 40 years or older. These individuals were invited to answer a questionnaire and undergo anthropometry, followed by prebronchodilator and postbronchodilator spirometry. We defined COPD as a ratio less than 0.7 of postbronchodilator forced expiratory volume in the first second over forced vital capacity. FINDINGS: Complete information, including spirometry, was obtained from 963 people in Sao Paulo, 1173 in Santiago, 1000 in Mexico City, 885 in Montevideo, and 1294 in Caracas. Crude rates of COPD ranged from 7.8% (78 of 1000; 95% CI 5.9-9.7) in Mexico City to 19.7% (174 of 885; 17.2-22.2) in Montevideo. After adjustment for key risk factors, the prevalence of COPD in Mexico City remained significantly lower than that in other cities. INTERPRETATION: These results suggest that COPD is a greater health problem in Latin America than previously realised. Altitude may explain part of the difference in prevalence. Given the high rates of tobacco use in the region, increasing public awareness of the burden of COPD is important. PMID- 16310555 TI - Economic assessment of a women's group intervention to improve birth outcomes in rural Nepal. AB - We did a cost-effectiveness analysis alongside a cluster-randomised controlled trial of a participatory intervention with women's groups to improve birth outcomes in rural Nepal. The average provider cost of the women's group intervention was US0.75 dollars per person per year (0.90 dollars with health service strengthening) in a population of 86,704. The incremental cost per life year saved (LYS) was 211 dollars (251 dollars), and expansion could rationalise on start-up costs and technical assistance, reducing the cost per LYS to 138 dollars (179 dollars). Sensitivity analysis showed a variation from 83 dollars to 263 dollars per LYS for most variables. This intervention could provide a cost effective way of reducing neonatal deaths. PMID- 16310556 TI - Prevention of falls and consequent injuries in elderly people. AB - Injuries resulting from falls in elderly people are a major public-health concern, representing one of the main causes of longstanding pain, functional impairment, disability, and death in this population. The problem is going to worsen, since the rates of such injuries seem to be rising in many areas, as is the number of elderly people in both the developed and developing world. Many methods and programmes to prevent such injuries already exist, including regular exercise, vitamin D and calcium supplementation, withdrawal of psychotropic medication, cataract surgery, professional environment hazard assessment and modification, hip protectors, and multifactorial preventive programmes for simultaneous assessment and reduction of many of the predisposing and situational risk factors. To receive broader-scale effectiveness, these programmes will need systematic implementation. Care must be taken, however, to rigorously select the right actions for those people most likely to benefit, such as vitamin D and calcium supplementation and hip protectors for elderly people living in institutions. PMID- 16310557 TI - The end or the beginning of the drive to an HIV-preventive vaccine: a view from over 20 years. PMID- 16310558 TI - What would Malthus say about AIDS in Africa? PMID- 16310559 TI - HIV transmission from surrogate breastfeeding. PMID- 16310560 TI - Urinary tract infections-microbial virulence determinants and reactive oxygen species. AB - The human urinary tract is able to combat with the microbial invasion under normal circumstances. To cause urinary tract infection the organism has to evade the host defense mechanisms by possessing distinct properties which contribute to the virulence of the organism hence called virulence determinants Ninety percent of uncomplicated urinary tract infections are caused by Escherichia coli, hence the knowledge of the virulence determinants of this organism can be extrapolated to other uropathogenic organism as well. Virulence determinants of uropathogenic E. coli include adhesins, siderophore production, polysaccharide coating, hemolysin production, outer membrane proteins etc. The intestinal E. coli, which are the reservoir of E. coli for causing UTI, lack these virulence determinants. On the other hand these virulence determinants enable the organism to colonize and invade the urinary tract. In addition these are important in acquiring the nutrients in other wise nutrient deficient environment. Further, they also help the organisms in triggering an inflammatory response and hence bringing about pathological changes which leads to symptomatic UTI. Severity of symptomatic infections and tissue damage during the infective process depends upon the magnitude of the inflammatory response triggered by the uropathogen which in turn is dependent upon the amount of extrcellular release of reactive oxygen species by the phagocytic cells; hence role of antioxidants as an adjunct to antibiotics in the treatment of infective process needs to be evaluated further. PMID- 16310561 TI - Pelvic insufficiency fracture related to autosomal dominant polycystic kidney disease. AB - We report the case of a patient with autosomal dominant polycystic kidney disease (ADPKD) and an insufficiency-type fracture of the pelvis. A 60-year-old Japanese woman was admitted because of pain in the right ischium and pubis that began suddenly with no precipitating cause. Computed tomography showed the bony pelvis to be compressed by enlarged dependent kidneys and an enlarged liver. We relieved compression on the pelvic bones by means of transarterial embolization (TAE) to the kidneys and liver after initiation of hemodialysis therapy. The fracture healed gradually after TAE, and the patient could walk 4 months later. In an iliac bone specimen obtained before TAE, cancellous bone was intact, but periosteal and endosteal surfaces of cortical bone showed marked resorption and were irregular. Normally, many ligaments are connected tightly to the periosteal surface, supporting the cortical bone. However, because of extensive surface resorption associated with pressure from enlarged kidneys, connections between ligaments and the periosteal surface presumably became fragile, promoting an insufficiency fracture from unapparent external forces. Thus, ADPKD is a potential cause of insufficiency fracture owing to abnormalities of cortical bone. PMID- 16310562 TI - Intracranial granuloma and skull osteolysis: complication of a primary cutaneous cryptococcosis in a kidney transplant recipient. AB - Cryptococcosis is the third most common invasive fungal infection in organ transplant recipients after candidiasis and aspergillosis. It occurs almost exclusively in the late posttransplantation period (>6 months after the initiation of immunosuppression). Subclinical onset of meningitis is the usual clinical presentation. Despite initiation of therapy, the mortality rate associated with this infection in this patient population remains high. To the best of our knowledge, this report describes one of the first cases of a rare entity: a primary cutaneous cryptococcosis in a renal transplant recipient disclosed by skull osteomyelitis and pseudotumoral intracranial extension. Surgical debridement and azole antifungal therapy were performed. Ten months after the onset of treatment, the patient feels good, clinical examination findings are normal, and no sign of evolutive cryptococcosis is noted. PMID- 16310563 TI - Peritonitis with multiple rare environmental bacteria in a patient receiving long term peritoneal dialysis. AB - We describe a patient receiving long-term peritoneal dialysis who experienced 2 episodes of peritonitis in successive months caused by unusual bacteria of environmental origin: Agrobacterium radiobacter, Pseudomonas oryzihabitans, and Corynebacterium aquaticum. A radiobacter and P oryzihabitans occurred simultaneously in the first episode of peritonitis, and C aquaticum, in the second episode. The patient's vocation necessitated exposure to moist soiled conditions. Both episodes responded promptly to antibiotics commonly used to treat peritonitis. Although these organisms rarely lead to loss of life and commonly are considered to be contaminants, they can cause symptomatic peritonitis and peritoneal dialysis catheter loss. A review of previous case reports is included. PMID- 16310564 TI - Acute renal failure induced by a Brazilian variety of propolis. AB - Propolis is a resinous substance collected by honeybees and used in hive construction and maintenance. Cumulative evidence suggests that propolis may have anti-inflammatory, antibiotic, antioxidant, antihepatotoxic, and antitumor properties. In addition to topical applications, products containing propolis have been used increasingly as dietary supplements. Although reports of allergic reactions are not uncommon, propolis is reputed to be relatively nontoxic. Its systemic toxicity is rarely reported and hence may be underestimated. This is the first report of propolis-induced acute renal failure. A 59-year-old man required hemodialysis for acute renal failure. The patient had cholangiocarcinoma and had ingested propolis for 2 weeks before presentation. Renal function improved after propolis withdrawal, deteriorated again after reexposure, and then returned to a normal level after the second propolis withdrawal. This case indicates that propolis can induce acute renal failure and emphasizes the need for vigilance and care when propolis is used as a medicine or dietary supplement. PMID- 16310566 TI - The use of vaccines in adult patients with renal disease. AB - In patients with renal disease, infection remains among the most common causes of morbidity and mortality. Alterations in the function of the immune system, as well as unique exposures of this patient population, account for the increased risk. Vaccination is an invaluable tool in preventing many infectious diseases. Unfortunately, responsiveness to vaccination in patients with renal disease can be diminished. In the present review, we examine the available evidence on the use of vaccinations in adult patients at different stages of chronic kidney disease. We address efficacy, clinical outcomes, and potential costs of individual vaccinations and provide our recommendations based on the literature reviewed. We also identify areas in which additional research is needed. PMID- 16310567 TI - Idiopathic membranous nephropathy: outline and rationale of a treatment strategy. AB - Idiopathic membranous nephropathy is a common cause of nephrotic syndrome. The treatment of patients with idiopathic membranous nephropathy is heavily debated. Based on literature data and our own experience, we propose a rational treatment strategy. Patients with renal insufficiency (serum creatinine level > 1.5 mg/dL [> 135 micromol/L]) are at greatest risk for the development of end-stage renal disease and should receive immunosuppressive therapy. In patients with normal renal function (serum creatinine level < 1.5 mg/dL [< 135 micromol/L]), risk for developing end-stage renal disease can be estimated by measuring urinary excretion of beta2-microglobulin or alpha1-microglobulin and immunoglobulin G. For low-risk patients, a wait-and-see policy is advised. High-risk patients likely benefit from immunosuppressive therapy. Currently, combinations of steroids with chlorambucil or cyclophosphamide are the best studied. We prefer cyclophosphamide in view of its fewer side effects. Cyclosporine may be an alternative option in patients with well-preserved renal function, although long term data are lacking. Other immunosuppressive agents, such as mycophenolate mofetil or rituximab, currently are under study; however, data are insufficient to support their routine use. PMID- 16310568 TI - Urinary tract infections in patients with urinary diversion. AB - Several surgical techniques have been used to provide urinary diversion after radical cystectomy. The noncontinent type of urinary diversion (using an intestinal conduit) and the continent urinary diversion (ureterosigmoidostomy or construction of an intestinal pouch that serves as a deposit of urine) were used commonly in the past, leading to a cumulative considerable number of patients encountered in clinical practice. The orthotopic urinary diversion (neobladder) has been used widely during the last years. The incidence and characteristics of urinary tract infection in patients with different types of urinary diversion are reviewed. Clinicians should be aware that urinary tract infection is a frequent cause of fever in patients with urinary diversion because it occurs in a significant proportion. In addition, symptoms from the lower urinary tract may not be prominent in this population. PMID- 16310569 TI - The outcome of acute renal failure in the intensive care unit according to RIFLE: model application, sensitivity, and predictability. AB - BACKGROUND: The definition, classification, and choice of management of acute renal failure (ARF) in the setting of the intensive care unit (ICU) remain subjects of debate. To improve our approach to ARF in the ICU setting, we retrospectively applied the new classification of ARF put forward by the Acute Dialysis Quality Initiative group, RIFLE (acronym indicating Risk of renal failure, Injury to the kidney, Failure of kidney function, Loss of kidney function, and End-stage renal failure), to evaluate its sensitivity and specificity to predict renal and patient outcomes. METHODS: RIFLE classification was applied to 183 patients with ARF admitted to the ICU (2002 to 2003) at the Northern General Hospital, Sheffield, UK. Patients were divided into 4 groups according to percentage of decrease in glomerular filtration rate from baseline. The risk group included 60 patients; injury group, 56 patients; failure group, 43 patients; and control group, 24 patients. Demographic, biochemical, hematologic, clinical, and long-term health status were studied and compared in the 4 groups. An attempt was made to evaluate, by means of logistic regression analysis and receiver operator characteristic curve analysis, the predictive value of RIFLE classification for mortality in the ICU. RESULTS: The failure group showed the worst parameters with regard to Acute Physiology and Chronic Health Evaluation (APACHE) II score, pH, lowest and highest mean arterial pressures, and Glasgow Coma Scale (P < 0.001). Mortality rate in the ICU (1 month) was significantly greater in the failure group compared with all groups (32 of 43 patients [74.4%]; P < 0.001) and, again, 6-month mortality rate (37 of 43 patients [86%]; P < 0.001). Receiver operator characteristic curve analysis showed that Simplified Acute Physiology Score (SAPS) II was more sensitive than APACHE II score for prediction of patient death in the risk and injury groups compared with the failure and control groups (risk group: SAPS II, 0.8 +/- 0.06; P < 0.001; APACHE II, 0.63 +/- 0.07; P = 0.14; injury group: SAPS II, 0.76 +/- 0.08; P < 0.001; APACHE II, 0.72 +/- 0.07; P = 0.006). CONCLUSION: RIFLE classification can improve the ability of such older and established ICU scoring systems as APACHE II and SAPS II in predicting outcome of ICU patients with ARF. PMID- 16310570 TI - Uncomplicated acute renal failure and hospital resource utilization: a retrospective multicenter analysis. AB - BACKGROUND: Although acute renal failure (ARF) complicating nonrenal organ dysfunction in the intensive care unit is associated with significant mortality and hospital costs, hospital resource utilization attributed to uncomplicated ARF is not well known. The goal of this study is to characterize the costs and lengths of stay (LOSs) incurred by hospitalized patients with uncomplicated ARF and their important determining factors. METHODS: We obtained hospital case-mix data sets from 23 Massachusetts hospitals for a 2-year period (1999 to 2000) from the Massachusetts Division of Health Care Finance and Policy. A total of 2,252 records of patients hospitalized with uncomplicated ARF were identified. Patient records of other common medical diagnoses were studied for comparison. RESULTS: Patients hospitalized with uncomplicated ARF incurred median direct hospital costs of 2,600 dollars, median hospital LOS of 5 days, and mortality of 8%. Dialysis was independently associated with significantly greater hospital costs and LOSs for patients with uncomplicated ARF (P < 0.05). Male sex and nonwhite race were associated with significantly lower hospital costs and LOSs, whereas type of hospital had opposing effects on these 2 resource utilization outcomes (P < 0.05). Unadjusted aggregate resource utilization associated with uncomplicated ARF exceeded that of many other common illnesses. CONCLUSION: Demographic and hospital factors, as well as dialysis therapy, are significant determinants of hospital resource utilization for patients with uncomplicated ARF. Uncomplicated ARF appears to incur greater hospital costs and longer LOSs compared with other common medical conditions. Greater focus should be directed toward further understanding of the factors influencing resource utilization for ARF. PMID- 16310571 TI - Sonographic assessment of the severity and progression of autosomal dominant polycystic kidney disease: the Consortium of Renal Imaging Studies in Polycystic Kidney Disease (CRISP). AB - BACKGROUND: The accuracy and precision of ultrasonography (US) in assessing the severity of autosomal dominant polycystic kidney disease (ADPKD) is unknown. METHODS: US and magnetic resonance imaging (MRI) were performed at baseline and 1 year on 230 subjects with ADPKD. Ellipsoid volume was calculated from US length, width, and depth, and sequential transverse images were used to measure total and cystic volume directly. These were compared with MRI measurements of kidney volume and cystic volume. RESULTS: Variability between different sonographers ranged from 18% to 42%. Correlations between US and MRI volume were 0.88 and 0.89. The SD of the discrepancy from MRI ranged from 21% to 33% and was unrelated to kidney size or body mass. Kidney length was the most reproducible measurement, and its correlation with MRI volume was 0.84. All patients with an US volume less than 700 cm3 had an MRI volume less than 1,000 cm3, and all patients with an US volume greater than 1,700 cm3 had an MRI volume greater than 1,000 cm3. Increases in volume after 1 year were 12% +/- 36% for the ellipsoid method, 6% +/- 29% for the direct method, and 4.2% +/- 7.2% for MRI. Correlation between US and MRI measurement of fractional cyst volume was 0.80. CONCLUSION: Sonographic measurement of kidney volume in patients with ADPKD is inaccurate and lacks the precision necessary to measure short-term disease progression. However, sonography can provide an estimate of kidney volume that reflects severity and prognosis in individual patients. PMID- 16310572 TI - Factors predicting outcome in a cohort of patients with atherosclerotic renal artery disease diagnosed by magnetic resonance angiography. AB - BACKGROUND: Atherosclerotic renal artery disease increasingly is recognized as a cause of chronic kidney disease and associated with high morbidity and mortality. We investigated factors predicting patient and renal survival in a cohort of patients with atherosclerotic renal artery disease diagnosed by means of magnetic resonance angiography (MRA). METHODS: We retrospectively analyzed a cohort of patients attending our unit in whom atherosclerotic renal artery disease was identified by means of MRA from 1998 to 2001. One hundred nine patients were followed up for a median of 2.3 years. Baseline clinical and laboratory data were assessed as predictors of outcome by using multivariate Cox proportional hazards analysis. RESULTS: Seventeen patients (16%) required dialysis and 37 patients (34%) died during a median follow-up of 841 days (interquartile range, 326 to 1,206). On multivariate Cox proportional hazards analysis, increased peripheral blood eosinophil count (hazard ratio, 3.39; 95% confidence interval [CI], 1.45 to 7.88; P = 0.0097), creatinine clearance (hazard ratio, 0.97; 95% CI, 0.94 to 0.99; P = 0.0128), and peripheral arterial disease (hazard ratio, 2.09; 95% CI, 1.04 to 4.18; P = 0.0371) were associated with subsequent death. Only creatinine clearance (hazard ratio, 0.91; 95% CI, 0.87 to 0.96; P = 0.0004) was associated with the need for dialysis. There was no association between eosinophil count and other markers of inflammation. Severity of atherosclerotic renal artery disease was not associated independently with either the need for dialysis or death. CONCLUSION: An increased peripheral-blood eosinophil count predicts patient survival in those with atherosclerotic renal artery disease at the time of diagnosis. This novel risk factor may help identify a group of patients who could benefit from intensive medical therapy by using an assay readily available to most clinicians worldwide. PMID- 16310573 TI - Long-term renoprotective effects of standard versus high doses of telmisartan in hypertensive nondiabetic nephropathies. AB - BACKGROUND: This report describes an open randomized study intended to evaluate the long-term renoprotective effects of "standard" (80 mg once daily) versus "high" (80 mg twice daily) doses of telmisartan in hypertensive patients without diabetes with biopsy-proven chronic proteinuric nephropathies. METHODS: We included 78 patients (age, 43.5 +/- 13.2 years; 71.8% men). After a 4-week wash out period, patients were randomly assigned to telmisartan, 80 mg once daily (n = 40) or 80 mg twice daily (n = 38), during a mean follow-up of 24.6 +/- 2.2 months. RESULTS: Baseline characteristics were similar in both groups, including blood pressure, renal function, and proteinuria. Blood pressure control did not differ between groups during follow-up. In the group administered telmisartan, 80 mg once daily, serum creatinine level increased from 1.6 +/- 0.6 to 2.7 +/- 0.9 mg/dL (141 +/- 52 to 239 +/- 80 micromol/L), and estimated creatinine clearance declined from 68 +/- 30 to 50 +/- 34 mL/min (1.13 +/- 0.50 to 0.83 +/- 0.57 mL/s), whereas in those administered 80 mg twice daily, serum creatinine (1.6 +/- 0.7 to 1.6 +/- 0.8 mg/dL [141 +/- 62 to 141 +/- 71 micromol/L]) and estimated creatinine clearance values (67 +/- 38 to 74 +/- 38 mL/min [1.12 +/- 0.63 to 1.23 +/- 0.63 mL/s]) did not change during the study. The decrease in proteinuria was more pronounced (P < 0.01) in patients administered the high dose of telmisartan compared with those treated with the standard dose. Serum potassium levels and lipid profiles did not change significantly in either group. CONCLUSION: Long term administration of high doses of telmisartan seems to improve the efficacy of the drug to decrease proteinuria and slow the progression to end-stage renal failure in nondiabetic hypertensive renal disease. PMID- 16310574 TI - Underuse of ACE inhibitors and angiotensin II receptor blockers in elderly patients with diabetes. AB - BACKGROUND: National guidelines recommend angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) therapy in patients with diabetes who also have hypertension and/or proteinuria to retard the progression of renal damage. However, little is known about the adequacy of adherence to these guidelines in elderly patients with diabetes and predictors of such appropriate ACE-inhibitor or ARB use. METHODS: Using linked medical claims from Medicare and the Pennsylvania Pharmaceutical Assistance Contract for the Elderly program, we studied a cohort of patients older than 65 years with diabetes. Baseline variables ascertained included age, sex, race, income, and several specific comorbid conditions. The outcome measure was at least 1 filled prescription for any ACE inhibitor or ARB during the quarter after the baseline year. We used multivariate logistic regression to measure predictors of use of the agents studied. RESULTS: Of 30,750 patients with diabetes studied, 21,138 patients (68.7%) also had hypertension and/or proteinuria. Of these, only 50.7% (95% confidence interval, 50.0 to 51.4) were administered an ACE inhibitor or ARB in the quarter studied. In multivariate analyses, failure to be administered either agent was associated significantly with older age, male sex, chronic lung disease, depression, dementia, and other mental illness. Greater rates of ACE inhibitor or ARB use were found in patients with coronary artery disease or congestive heart failure. CONCLUSION: Only half the elderly patients with diabetes studied received recommended treatment with ACE inhibitors or ARBs. This shortfall could provide an opportunity for quality-improvement interventions that could result in important benefits in clinical outcomes for these high-risk patients. PMID- 16310575 TI - Predialysis psychoeducational intervention extends survival in CKD: a 20-year follow-up. AB - BACKGROUND: Predialysis psychoeducational interventions increase patient knowledge about chronic kidney disease (CKD) and its treatment and extend time to dialysis therapy without compromising physical well-being in the short run. The present research examines long-term survival after predialysis psychoeducational intervention. In addition, we examined whether survival differed because of early (ie, > or = 3 months) versus late referral to nephrology. METHODS: We collected follow-up data for patients with CKD who participated in a multicenter randomized controlled trial of predialysis psychoeducational interventions in the mid-1980s. We gathered 20-year survival data from clinical records and databases. RESULTS: Participants included 335 patients with CKD, including 172 patients randomly assigned to receive predialysis psychoeducational interventions (63.0% men; mean age, 50.8 years) and 163 patients assigned to usual care (62.1% men; mean age, 52.7 years). Two hundred forty-six patients (66.8%) died during the course of the study. Mean duration of follow-up was 8.5 +/- 7.23 (SD) years. Analyses were by intention to treat. Adjusting for age, general nonrenal health at inception, and time between identification and predialysis psychoeducational intervention or usual care, Cox proportional hazards multiple regression analyses indicated that median survival was 2.25 years longer after patients with CKD received predialysis psychoeducational interventions compared with usual care (chi-square change [1] = 3.75; P = 0.053; hazard ratio, 1.32; 95% confidence interval, 1.0 to 1.74). Predialysis psychoeducational intervention recipients survived a median of 8.0 months longer than usual-care patients after the initiation of dialysis therapy (chi-square-change [1] = 4.39; P = 0.036; hazard ratio, 1.35; 95% confidence interval, 1.02 to 1.775). No significant survival advantage was evident for early referral to nephrology or the combination of early referral plus predialysis psychoeducational interventions. CONCLUSION: Predialysis psychoeducational intervention is a safe and useful intervention that contributes valuably to multidisciplinary predialysis care. PMID- 16310576 TI - Variable effects of soy protein on plasma lipids in hyperlipidemic and normolipidemic hemodialysis patients. AB - BACKGROUND: Hyperlipidemic factors contribute to the high cardiovascular risk in hemodialysis patients. Soy protein has decreased some atherogenic lipid concentrations in subjects with normal renal function. This study evaluates the effect of soy protein on serum lipid profiles in hyperlipidemic and normolipidemic hemodialysis patients. METHODS: Nineteen hyperlipidemic and 18 normolipidemic hemodialysis patients were enrolled in a randomized, double-blind, placebo-controlled, clinical trial. After a 4-week run-in phase, subjects in each category were randomly assigned to 2 groups. Thirty grams of isolated soy protein or milk protein was consumed daily as a beverage at breakfast or postdialysis for 12 weeks. RESULTS: In hyperlipidemic subjects, soy protein intake significantly decreased total cholesterol levels by 18.6% (95% confidence interval [CI], -11.4 to -25.8; P = 0.04), triglyceride levels by 43.1% (95% CI, -34.0 to -52.2; P = 0.02), non-high-density lipoprotein cholesterol levels by 23.6% (95% CI, -14.7 to -32.5; P < 0.01), apolipoprotein B levels by 15.4% (95% CI, -5.4 to -25.4; P = 0.01), and insulin levels by 49.8% (95% CI, -23.3 to -66.1; P < 0.01). Low density lipoprotein cholesterol concentration was decreased significantly ( 25.8%; 95% CI, -8.3 to -42.7; P = 0.01), and high-density lipoprotein cholesterol level was increased significantly (17%; 95% CI, 2 to 32.0; P = 0.03), but there was no significant difference compared with the milk protein group (-5.5% +/- 16.9% and 7.0% +/- 11.8%, respectively). There were no significant changes in serum lipid and lipoprotein concentrations in normolipidemic subjects. CONCLUSION: These results indicate soy protein substitution has lipid-lowering effects in hyperlipidemic hemodialysis patients. However, soy protein intake had little effect on plasma lipid levels in normolipidemic hemodialysis patients. PMID- 16310577 TI - Skipped treatments, markers of nutritional nonadherence, and survival among incident hemodialysis patients. AB - BACKGROUND: Skipping hemodialysis treatments and failing to adhere to prescribed diets are thought to injure hemodialysis patients. METHODS: We examined predictors of hemodialysis skipping and laboratory measures of nonadherence and then examined the association of dialysis skipping and serum potassium and phosphate levels with survival. RESULTS: Of 739 patients, 67 were classified as skippers because they were absent for greater than 3% of scheduled treatments. Black race (odds ratio [OR], 2.26; 95% confidence interval [CI], 1.30 to 3.92), current smoking (OR, 1.79; 95% CI, 1.02 to 3.13), and use of illicit drugs (OR, 3.96; 95% CI, 2.16 to 7.24) were associated with skipping. White race, increased serum phosphate level, greater creatinine level, and lower body mass index increased the likelihood of a serum potassium level greater than 5.0 mEq/L (mmol/L); younger age, greater serum potassium level, and greater serum creatinine level were associated with a serum phosphate level greater than 5.5 mg/dL (>1.78 mmol/L). Skipping was associated with an increased risk for death (hazard ratio [HR], 1.69; 95% CI, 1.24 to 2.31), as were phosphate level greater than 5.5 mg/dL (HR, 1.59; 95% CI, 1.16 to 2.17) and potassium level greater than 5.0 mEq/L (HR, 1.50; 95% CI, 1.10 to 2.06). Skipping was associated with a lower likelihood of kidney transplantation in those younger than 65 years (OR, 0.41; 95% CI, 0.18 to 0.93). CONCLUSION: These findings show that hemodialysis patients of black race and those with current tobacco or illicit drug use are at risk for skipping dialysis treatments. Skipping treatments and markers of poor dietary adherence are strongly associated with greater risk for death. Targeting high risk patients to understand reasons for nonadherence and to intervene could prevent premature death. PMID- 16310578 TI - Wait-listing for kidney transplantation among Aboriginal hemodialysis patients. AB - BACKGROUND: Although there has been rapid growth in the global prevalence of Aboriginal patients with end-stage renal disease (ESRD), these individuals have markedly lower rates of kidney transplantation for reasons that are unclear. We investigated barriers to kidney transplantation for Aboriginal patients treated with hemodialysis for ESRD in Canada. METHODS: All consenting adults in the province of Alberta, Canada, who had been established on hemodialysis therapy for 6 months or longer were interviewed by a physician. Data for transplantation referral, waiting list status, and the assessment process were determined from the regional transplant programs, with race defined by patient self-report. For purposes of this analysis, race was dichotomized as either Aboriginal or non Aboriginal. RESULTS: Of 835 subjects, 100 (12%) were Aboriginal. Aboriginal patients were significantly younger and more likely to have diabetes mellitus as the cause of ESRD than non-Aboriginal patients. Although a greater proportion of Aboriginal patients were referred for transplantation assessment (60.6% versus 46.0%; P < 0.01), after adjustment, the likelihood of referral was similar for both racial groups (hazard ratio associated with Aboriginal race, 0.80; 95% confidence interval, 0.59 to 1.08). Aboriginal patients also were significantly less likely to be active on the transplant wait list than non-Aboriginal patients (adjusted hazard ratio, 0.46; 95% confidence interval, 0.27 to 0.78). Aboriginal subjects who had been referred for assessment were significantly more likely than non-Aboriginals to be in the process of completing the transplantation workup (69.6% versus 26.9%; P < 0.01) as opposed to being temporarily or permanently unsuitable for transplantation (30.4% versus 73.3%; P < 0.01). Although not statistically significant, median duration of the workup in progress in referred, but nonlisted, participants appeared longer in Aboriginal participants (954 versus 596 days; P = 0.07). CONCLUSION: The likelihood of referral for transplantation was similar between Aboriginal and non-Aboriginal people. However, Aboriginal people were approximately half as likely to be successfully activated to the transplant waiting list compared with non-Aboriginal people. These data suggest that the major barrier to transplantation in Aboriginal patients occurs after referral, but early in the course of evaluation for eligibility. PMID- 16310579 TI - Thrombotic microangiopathy in marginal kidneys after sirolimus use. AB - The increasing shortage of cadaver donor kidneys has prompted the use of expanded or marginal donor kidneys, ie, from older donors or those with a history of hypertension or diabetes. These marginal kidneys may be especially susceptible to calcineurin inhibitor (CNI)-mediated vasoconstriction and nephrotoxicity. Recipients of renal transplants from marginal donors therefore require non nephrotoxic immunosuppression. Some investigators have proposed using sirolimus, a novel and potent immunosuppressant, instead of CNIs. Moreover, another complication of solid-organ transplantation is thrombotic microangiopathy (TMA), which affects 3% to 14% of patients on immunosuppression therapy treated with CNIs. Therefore, it was suggested that CNIs may be substituted by sirolimus in patients with posttransplantation CNI-induced TMA. We report 3 patients who received marginal cadaveric kidneys and were administered maintenance immunosuppression with sirolimus, prednisone, and mycophenolate mofetil. They each developed de novo TMA despite never having been previously administered a CNI. In these cases, TMA occurred in marginal kidneys with possible endothelial injury before transplantation. Sirolimus may have prevented recovery from these injuries and thus may have promoted TMA in these marginal kidneys. The risk for such a vascular complication should be kept in mind in patients who receive marginal kidneys and are administered sirolimus, even when sirolimus is used without CNIs. PMID- 16310580 TI - Pharmacology. PMID- 16310581 TI - Purpura followed by proteinuria in a 7-year-old girl. PMID- 16310583 TI - Prevalence of left ventricular hypertrophy: some alternate thoughts. PMID- 16310584 TI - Cardiovascular mortality and dialysis access-related infections: is there a link? PMID- 16310585 TI - A water deprivation test is not indicated in the evaluation of hypernatremia. PMID- 16310587 TI - The neurology of attention deficit/hyperactivity disorder. PMID- 16310588 TI - Apolipoprotein E genotype analysis in Chinese Han ethnic children with Wilson's disease, with a concentration on those homozygous for R778L. AB - Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism caused by a large number of different mutations in the ATP7B gene. R778L mutation is mostly observed in Chinese, Japanese and Korean patients, whereas the H1069Q point mutation in the ATP7B gene is the most frequent mutation in European patients with WD. In our previous study we did not find a significant correlation between genotype and phenotype (age of onset and clinical presentation) in patients homozygous (37 patients) or heterozygous (52 patients) for R778L. It was reported that European patients homozygous for H1069Q who were also homozygous for the ApoE genotype epsilon3/3 developed clinical symptoms 5-11 years later than did patients with genotypes other than ApoE epsilon3/3. In the present study (i) we firstly observed that ApoE epsilon3/3 did not delay the onset of WD; (ii) no association between ApoE genotype and WD clinical presentation in Chinese Han children, including those patients homozygous for R778L. Thus we conclude that the onset of WD in Chinese children is not related to ApoE epsilon3/3, although the high frequency of ApoE epsilon3/3 in Chinese Han children with WD was not significantly different from that in controls. PMID- 16310589 TI - Low-dose carbamazepine therapy for benign infantile convulsions. AB - Benign infantile convulsions (BIC) are characterized by: (1) onset at up to 2 years of age, (2) normal development, (3) mostly brief, often clustered convulsions, and (4) normal electroencephalography during the interictal stage. BIC follow a favorable course and disappear before 2-3 years of age, although convulsions for which diazepam is ineffective frequently develop. We treated 15 children (3-16 months of age) diagnosed as having BIC, excluding convulsions associated with mild gastroenteritis, with a once-daily dose of 5mg/kg of carbamazepine until up to 2 or 3 years of age. The serum concentration of carbamazepine was as low as below the effective range in six patients, but the treatment was dramatically effective in all the BIC children. Seizures did not recur in any patients during oral administration of carbamazepine. The treatment was finished in 12 patients at age 2 years, two at age 3 years, and one at 16 months-old. Therefore, we recommend the administration of a once-daily dose of 5mg/kg of carbamazepine until up to 2 or 3 years of age as a treatment for BIC. PMID- 16310590 TI - Synchronous occurrence of EEG bursts and heart rate acceleration in preterm infants. AB - Continuous and simultaneous registration of electroencephalogram (EEG) and heart rate (HR) pattern in preterm infants can give information about the functioning of central nervous system and the integrity of the autonomic nervous system. The developmental and behavioural state determine the pattern of EEG activity. A discontinuous EEG activity also known as 'Trace alternant' (TA) in preterm infants is accompanied by a low heart rate variability (HRV). It was found that electroencephalographic bursts of slow waves during TA are coupled with an acceleration of the HR. In this study, this synchronous behaviour of EEG bursts and HR is described for the first time in a group of preterm infants with a mean conceptional age (CA) of 36 weeks. PMID- 16310591 TI - The Rey-Osterrieth Complex Figure as a measure of executive function in childhood. AB - In adults, the Rey-Osterrieth Complex Figure (ROCF) can be used for the assessment of not only visuoconstructional ability and visual memory, but also executive function in adults. We studied whether ROCF scores also correlated with executive function in childhood. The subjects consisted of 56 patients with various neurological diseases (5 years 7 months-14 years 11 months; mean: 8 years 8 months; M 42, F 14) whose full-scale IQs were 70 or higher (mean: 93.3). All subjects underwent the Wechsler Intelligence Scale for children third edition (WISC-III) and various psychological tests focusing on executive function. We evaluated the ROCF with the Boston Qualitative Scoring System (BQSS). We calculated the partial correlation coefficients between the BQSS Summary Scores and representative scores of other executive function tests, using age as the control variable. Among the scores of the various examinations, the Perseverative Errors of Nelson of the Wisconsin Card Sorting Test, the raw scores of the Mazes (WISC-III), the Digit Span (WISC-III), and the Block Design (WISC-III) were correlated significantly with two to four BQSS Summary Scores (P<0.05 or P<0.01). In contrast, the scores of the Trail Making Test, the Stroop Test, and the Commission Error of Continuous Performance Test-II did not show a significant correlation with any of the BQSS Summary Scores. The ROCF evaluated with the BQSS reflects not only visuoperceptual ability and visuoconstructional ability, but also executive function, especially planning and organization. However, a new BQSS Summary Score should be devised since no existing BQSS index specifically reflects executive functions with an outstandingly strong correlation. PMID- 16310592 TI - L-carnitine protects against glutamate- and kainic acid-induced neurotoxicity in cerebellar granular cell culture of rats. AB - Glutamate mediated intracellular calcium accumulation and free radical generation are thought to be major mechanisms that contribute to cell death in hypoxic ischemic brain injury. For this reason, various glutamate receptor antagonists and antioxidants have been investigated for their therapeutic potential. To assess whether L-carnitine, a possible antioxidant, is able to prevent glutamate- and kainic acid (KA)-induced neurotoxicity. Glutamate (10(-7) M) and one of its receptor agonists, KA (10(-4) M) were administered to cerebellar granular cell cultures that were prepared from 1-day-old Sprague-Dawley rats. The neuroprotective effect of L-carnitine was examined. L-carnitine at doses of 10( 6), 10(-5), 10(-4), 10(-3) M was applied to culture flasks. L-carnitine at doses of 10(-4) and 10(-3) M significantly blocked glutamate-induced neurotoxicity. 10( 4) M dose of L-carnitine proved to be more effective than 10(-3)M. L-carnitine also blocked KA-induced neurotoxicity only at the dose of 10(-4) M. 10(-4) M L carnitine, the most effective dose in both glutamate- and KA-induced neurotoxicity, decreased glutamate-induced neuronal cell death from 36.14+/-2.95% to 17.59+/-2.25%; (P<0.001) and KA-induced neuronal cell death from 21.4+/-0.41 to 13.4+/-1.38%; (P<0.001). The present study demonstrates that L-carnitine protects against glutamate- and KA-induced neurotoxicity. Protective effect of L carnitine may result from its antioxidant activity because free radical generation is a common result in either glutamate- or KA-induced neurotoxicity. L carnitine merits further investigation as a therapeutic option in hypoxic ischemic brain injury of newborn. PMID- 16310593 TI - Early onset distal muscular dystrophy with normal dysferlin expression. AB - A 7-year-old boy, who was noted to be a slow runner at the age of 2 years, had progressive muscle weakness and atrophy, preferentially affecting distal muscles. At 3 years of age, he had scoliosis and difficulty in standing on tip-toe. Serum creatine kinase was 1074IU/l. Muscle CT scan showed low-density areas in the lower legs and upper arms, but predominantly in the gastrocnemius and soleus muscles. Biopsy of the biceps brachii muscle showed moderate dystrophic changes with normal dysferlin expression on immunohistochemical and western blot analyses. Although muscle involvement mimicked that seen in Miyoshi myopathy (MM), the very early onset of the disease and scoliosis were quite unusual for MM. We, therefore, made the diagnosis of early onset dysferlin-positive distal muscular dystrophy, probably a new type of distal muscular dystrophy. PMID- 16310594 TI - Vitamin B12 deficiency in infancy as a cause of developmental regression. AB - Vitamin B12 deficiency can cause serious developmental regression, hypotonia and cerebral atrophy in infants. We report a 6-month-old infant, with insidious developmental regression and brain atrophy showed by CT scan, secondarily to vitamin B12 deficiency. His mother was a strict vegetarian and the patient was exclusively breastfed. The clinical symptoms and the brain CT were normalized after vitamin B12 administration. PMID- 16310595 TI - Dystonia and multiple sclerosis. PMID- 16310597 TI - Deep vein thrombosis in the intensive care unit: underappreciated, understudied, and undertreated. PMID- 16310598 TI - Preventing venous thromboembolism in the intensive care unit: A no-brainer? PMID- 16310599 TI - On the need for a workshop on venous thromboembolism in critical care. PMID- 16310600 TI - Prevalence, incidence, and risk factors for venous thromboembolism in medical surgical intensive care unit patients. PMID- 16310601 TI - The diagnosis of deep venous thrombosis and pulmonary embolism in medical surgical intensive care unit patients. PMID- 16310602 TI - Thromboprophylaxis in medical-surgical intensive care unit patients. PMID- 16310603 TI - Anticoagulant prophylaxis in special populations with an indwelling epidural catheter or renal insufficiency. AB - The prevention of deep venous thrombosis in postoperative patients who have an indwelling epidural catheter and patients with impaired renal function is a frequently encountered and challenging clinical management problem. Such patients make up a considerable proportion of patients in a medical-surgical intensive care unit. The management of such patients is problematic because low-molecular weight heparins, the anticoagulant of choice as thromboprophylaxis for many clinical indications, are generally avoided in such patients. This review focuses on anticoagulant prophylaxis in such patients, including reasonable approaches for using low-molecular-weight heparins as thromboprophylaxis. PMID- 16310604 TI - Deep venous thrombosis: clinically silent in the intensive care unit. AB - PURPOSE: The reliability of clinical signs and the physical examination in the evaluation of deep venous thrombosis (DVT) in the critically ill is unknown. The purpose of this study was to determine the diagnostic properties of clinical examination for signs of DVT in a cohort of medical-surgical intensive care unit (ICU) patients using screening compression ultrasonography as a reference standard. MATERIALS AND METHODS: We prospectively included patients older than 18 years with an expected length of ICU stay of more than 72 hours. Patients underwent bilateral lower limb screening compression ultrasound twice weekly and structured physical examination twice weekly by 2 independent trained research coordinators blinded to the ultrasonography results. We classified patients according to 2 methods: method 1, a DVT Risk Stratification System of 3 categories and method 2, a DVT Risk Score, both of which use the history and physical examination to stratify patients for their risk of DVT. RESULTS: We included 239 patients in our study, 32 of whom had DVT based on the results of their compression ultrasound. We excluded 7 patients with DVT on ICU admission and 2 who did not undergo any structured examinations. We matched controls with cases (9:1) based on duration of ICU stay. Cases and controls were then allocated to low, moderate, and high risk strata for DVT. Using method 1, the area under the receiver operating characteristic curve (AUC) was 0.57 (95% CI, 0.33-0.78, P = .01). Using method 2, the AUC was 0.59 (95% CI, 0.42-0.75, P = .02). An AUC of 1.0 indicates an ideal test, and AUC of 0.50 indicates a test with no diagnostic utility. CONCLUSIONS: The history and physical examination for DVT are not useful in detecting lower limb DVT in the ICU. PMID- 16310605 TI - Burden of illness in venous thromboembolism in critical care: a multicenter observational study. AB - PURPOSE: The frequency of clinically diagnosed venous thromboembolism (VTE) including deep venous thrombosis (DVT) and pulmonary embolism (PE) in medical surgical critically ill patients is unclear. The objectives of this study were to estimate the prevalence and incidence of radiologically confirmed DVT and PE in medical-surgical intensive care unit (ICU) patients and to determine the impact of prophylaxis on the frequency of these events. MATERIALS AND METHODS: In a retrospective observational cohort study in 12 adult ICUs, we identified prevalent cases (diagnosed in the 24 hours preceding ICU admission up to 48 hours post-ICU admission) and incident cases (diagnosed 48 hours or more after ICU admission and up to 8 weeks after ICU discharge) of upper or lower limb DVT or PE. Deep venous thrombosis was diagnosed by compression ultrasound or venogram. Each DVT was classified as clinically suspected or not clinically suspected in that the latter was diagnosed by scheduled screening ultrasonography. Pulmonary embolism was diagnosed by ventilation-perfusion lung scan, computed tomography pulmonary angiography, echocardiography, electrocardiography, or autopsy. RESULTS: Among 12,338 patients, 252 (2.0%) patients had radiologically confirmed DVT or PE and another 47 (0.4%) had possible DVT or PE. Prevalent DVTs were diagnosed in 0.4% (95% confidence interval [CI], 0.3%-0.5%) of patients and prevalent PEs were diagnosed in 0.4% (95% CI, 0.3%-0.6%). Incident DVTs were diagnosed in 1.0% (95% CI, 0.8%-1.2%) of patients, and incident PEs were diagnosed in 0.5% (95% CI, 0.4%-0.6%). Of patients with incident VTE, 65.8% of cases occurred despite receipt of thromboprophylaxis for at least 80% of their days in ICU. The median (interquartile range) ICU length of stay was similar for patients with DVT (7 [3-17]) and PE (5 [2-8]). For all patients with VTE, ICU mortality was 16.7% (95% CI, 12.0%-21.3%) and hospital mortality was 28.5% (95% CI, 22.8%-34.1%). CONCLUSIONS: Venous thromboembolism appears to be an apparently infrequent, but likely underdiagnosed problem, occurring among patients receiving prophylaxis. Findings suggest the need for increased suspicion among clinicians, renewed efforts at thromboprophylaxis, and evaluation of superior prevention strategies. PMID- 16310606 TI - Thrombocytopenia in medical-surgical critically ill patients: prevalence, incidence, and risk factors. AB - OBJECTIVE: The purpose of this study is to describe the prevalence, to analyze the incidence and independent risk factors for thrombocytopenia, and to examine the impact of thrombocytopenia developing in the intensive care unit (ICU) on patient outcome in a well-defined cohort of critically ill patients in a medical surgical ICU. MATERIALS AND METHODS: As part of a prospective cohort study examining the frequency and clinical importance of venous thromboembolism in the ICU, we enrolled consecutive patients older than 18 years expected to be in the ICU for more than 72 hours. Exclusion criteria were an admitting diagnosis of trauma, orthopedic surgery or cardiac surgery, pregnancy, and life support withdrawal. Patients had platelet counts performed as directed by clinical need. We defined thrombocytopenia as a platelet count of less than 150 x 10(9)/L and severe thrombocytopenia as a platelet count of less than 50 x 10(9)/L. Protocol directed care included routine thromboprophylaxis and twice weekly screening ultrasonography of the legs. Patients were followed to hospital discharge. RESULTS: Of the 261 enrolled patients, 121 (46%, 95% confidence interval [CI], 40%-53%) had thrombocytopenia (62 on ICU admission and 59 acquired during their ICU stay). Patients who developed a platelet count less than 150 x 10(9)/L during their ICU stay had higher ICU and hospital mortality (P = .03 and .005, respectively), required longer mechanical ventilation (P = .05), and were more likely to receive platelets (P < .001), fresh frozen plasma (P = .005), and red blood cell transfusions (P = .004) than patients who did not develop thrombocytopenia. The only independent risk factors for thrombocytopenia developing during the ICU stay were administration of nonsteroidal anti inflammatory drugs before ICU admission (hazard ratio, 2.8; 95% CI, 1.3-6.0) and dialysis during the ICU stay (hazard ratio, 3.1; 95% CI, 1.2-7.8). Of the 33 patients who underwent 36 tests for heparin-induced thrombocytopenia, none tested positive. CONCLUSIONS: We found that about 50% of the patients admitted to the ICU had at least one platelet count of less than 150 x 10(9)/L during their ICU stay. Patients who developed thrombocytopenia were more likely to die, required longer duration of mechanical ventilation, and were more likely to require blood product transfusion. Heparin-induced thrombocytopenia was frequently suspected but did not develop in these critically ill patients. PMID- 16310607 TI - Thromboprophylaxis for intensive care patients in Australia and New Zealand: a brief survey report. AB - PURPOSE: Internationally, there is practice variation concerning optimal thromboprophylaxis for patients in the intensive care unit (ICU). The current practice in Australia and New Zealand is unknown. METHOD: We conducted a self administered e-mail survey of 22 Australian and New Zealand ICUs expressing interest in participating in a proposed international randomized trial (PROphylaxis for ThromboEmbolism in Critical Care Trial). RESULTS: Our response rate was 95.4% (95% CI, 77%-100%). Of participating ICUs, 90.5% (95% CI, 70%-99%) used subcutaneous unfractionated heparin for routine thromboprophylaxis in ICU patients. Low-molecular-weight heparin was reserved for specific high-risk patients in many units. CONCLUSION: Routine thromboprophylaxis for ICU patients in Australia and New Zealand is similar to Canada but different to France. Optimal thromboprophylaxis for ICU patients is currently unclear in the absence of randomized trial data. PMID- 16310608 TI - Dalteparin thromboprophylaxis for critically ill medical-surgical patients with renal insufficiency. AB - PURPOSE: Thromboprophylaxis with low-molecular-weight heparin (LMWH) may be more effective than unfractionated heparin but also more likely to bioaccumulate and potentially cause bleeding in patients with renal insufficiency. The objectives of this study were to assess, among medical-surgical patients in the intensive care unit receiving dalteparin 5,000 IU daily for thromboprophylaxis, (1) the relationship between renal dysfunction and LMWH bioaccumulation as measured by trough anti-Xa levels, (2) the relationship between renal dysfunction and risk of bleeding as measured by a surrogate marker (peak anti-Xa levels), and (3) the relationship between anti-Xa levels, bleeding events, and thrombotic events. MATERIALS AND METHODS: In this prospective single-center cohort study, we enrolled patients 18 years or older, expected to stay 72 hours or longer, and with a creatinine clearance 30 mL/min or higher at intensive care unit admission. We administered 5,000 IU dalteparin subcutaneously each day. The main phase 1 objective was to detect bioaccumulation of dalteparin by measuring trough anti-Xa levels (22-23 hours post dalteparin). The main phase 2 objective was to examine the relationship between renal dysfunction and peak anti-Xa levels (4 hours post dalteparin). We recorded creatinine clearance daily and bleeding and thrombotic events, blinded to anti-Xa levels. RESULTS: We enrolled 19 patients aged 62.7 (13.2) years with an APACHE II score of 23.5 (9.4). We measured trough anti-Xa levels on 185 occasions in 19 patients; we measured peak anti-Xa levels on 113 occasions in 11 patients. We identified no bioaccumulation of LMWH in this study, as detected by trough anti-Xa levels. Most peak anti-Xa levels were in the conventional prophylactic range. CONCLUSIONS: When administered in prophylactic doses to critically ill patients with a wide range of calculated creatinine clearances, we found no evidence of bioaccumulation of dalteparin. If dalteparin does not bioaccumulate, it may be an attractive alternative agent for thromboprophylaxis. PMID- 16310609 TI - Prophylaxis of Thromboembolism in Critical Care (PROTECT) Trial: a pilot study. AB - PURPOSE: There is no randomized trial comparing low-molecular weight heparin (LMWH) and unfractionated heparin (UFH) for thromboprophylaxis in medical surgical ICU patients. The primary objective of this randomized pilot study on LMWH vs UFH was to assess the feasibility of conducting a large randomized trial with respect to timely enrollment and blinded study drug administration, practicality of twice-weekly lower limb ultrasounds to screen for deep venous thrombosis, LMWH bioaccumulation and dose adjustment in renal insufficiency, and recruitment rates for a future trial in medical-surgical intensive care unit (ICU) patients. Its additional goals were to evaluate the suitability of the exclusion criteria and to document the range of research activities that precede accrual of patients into a trial to plan multisite management. MATERIALS AND METHODS: By computerized telephone randomization, we allocated 129 medical surgical ICU patients to treatment with dalteparin 5,000 IU QD SC or that with UFH 5,000 IU BID SC. Within each clinical center, only the study pharmacist was not blinded. We performed bilateral lower limb compression ultrasounds within 48 hours of ICU admission, twice weekly, on suspicion of deep venous thrombosis, and 7 days after ICU discharge. Research coordinators and investigators at 7 centers reported the time they engaged in all research activities before the first patient was randomized. RESULTS: Timely complete study drug administration occurred after enrollment. More than 99% of scheduled doses were administered in a blinded fashion. Scheduled ultrasounds were performed without exception. No bioaccumulation of dalteparin was observed when creatinine clearance decreased to lower than 30 mL/min. Average recruitment was 2 patients/center per month before the study exclusion criteria were modified. Study startup activities required, on average, 65.5 hours of combined investigator and research coordinator time at each center. Careful examination of the accrual in the pilot study led to a reexamination of the Prophylaxis of Thromboembolism in Critical Care Trial (PROTECT) study exclusion criteria. CONCLUSIONS: This pilot study suggests that a multicenter randomized clinical trial comparing LMWH with UFH in critically ill medical-surgical patients is feasible. Pilot studies can improve the design of larger trials and may enhance successful timely completion. PMID- 16310610 TI - Clinically important venous thromboembolism in pediatric critical care: a Canadian survey. AB - PURPOSE: Pediatric venous thromboembolism (VTE) is becoming an increasingly recognized morbidity associated with critical illness. The objective of this survey is to identify the patient factors and radiological features that pediatric intensivists consider more or less likely to make a venous thrombosis (VTE) clinically important in their patients. MATERIALS AND METHODS: Our definition of clinically important VTE was a VTE likely to result in short- or long-term morbidity or mortality if left untreated. We asked respondents to rate the likelihood that patient factors and radiological features make a venous thrombosis clinically important using a 5-point scale (1 = much less likely to 5 = much more likely). RESULTS: The 38 (58.5%) of 65 pediatric intensivists responding rated 4 patient factors as most likely to make a VTE clinically important: clinical suspicion of pulmonary embolism (mean score, 4.8), symptoms (mean, 4.5), detection by physical exam (mean, 4.4), and the presence of an acute or chronic cardiopulmonary comorbidity that might limit a patient's ability to tolerate pulmonary embolism (mean, 4.3). Of the radiological features, the 2 considered most important were VTE involving the vena cava extending into the right atrium (mean, 5) and central veins (mean, 4.5). CONCLUSIONS: When labeling a VTE as clinically important, pediatric intensivists rely on several specific patient factors and thrombus characteristics. PMID- 16310611 TI - Systems biology and translational research. PMID- 16310612 TI - Oral transmucosal fentanyl citrate in the management of dyspnea crises in cancer patients. PMID- 16310613 TI - Successful use of ketamine for intractable burning pain of HTLV-1-associated myelopathy. PMID- 16310614 TI - Late referral to hospice and bereaved family member perception of quality of end of-life care. AB - The Family Evaluation of Hospice Services was used to document bereaved family members' perceptions of whether their loved ones were referred too late to hospice and to examine the association of that perception and quality of end-of life care. A mortality follow-back survey of bereaved family members from two not for-profit hospices found that 13.7% of decedents were referred at a time too late for hospice services. Family members of persons referred too late reported lower satisfaction with hospice services, a higher rate of unmet needs for information about what to expect at time of death, lower confidence in participating in patient care at home, more concerns with coordination of care, and lower overall satisfaction. Families reported physicians as an important barrier to earlier hospice referral in nearly one-half of cases. These results indicate a need for improved services for shorter-stay hospice patients/families and for physicians to help facilitate earlier hospice admission. PMID- 16310615 TI - Patient reports of symptoms and their treatment at three palliative care projects servicing individuals with HIV/AIDS. AB - Self-reports of 32 symptoms and their treatments were obtained from patients of three palliative care programs that provide services to seriously ill HIV patients (>or=95% AIDS) in Alabama (n=47), Baltimore (n=91), and New York City (n=117). On average, patients reported 10.9 (SD=7.6) to 12.7 (SD=6.2) symptoms. Pain, lack of energy, and worrying were reported by a majority of patients at all sites, often with a high level of associated distress. For only four symptoms (pain, nausea, difficulty swallowing, and mouth sores) did half or more of patients at all sites experiencing the symptom also report treatment. Less than a third of patients experiencing 12 symptoms (five of six comprising a psychological subscale) reported treatment. Results show that despite the availability of more efficacious treatments, many HIV/AIDS patients continue to experience significant physical and psychological symptomatology. Many of those experiencing symptoms, however, do not perceive their symptoms as being treated. PMID- 16310617 TI - Occurrence, severity, and longitudinal course of twelve common symptoms in 1129 consecutive patients during radiotherapy for cancer. AB - Little is known about the frequency, severity, and course of symptoms experienced by patients receiving radiotherapy (RT). For this descriptive study, 1129 patients with a variety of cancer diagnoses completed a 12-item Symptom Inventory (SI) at the start of RT; 419 of these patients also completed the SI weekly for an additional 4 weeks (five data points). Eighty-four percent of the 1129 patients were already experiencing symptoms when treatment began. All symptoms significantly increased in frequency over a typical 5 week RT course (all Ps<0.001). Skin problems showed the largest increase. The most common symptoms (fatigue, drowsiness, and sleep problems) were also the most severe. Female patients and patients younger than the median age (59 years) reported significantly more symptoms than males and those 59 years or older. Symptom frequency and severity varied significantly by cancer diagnosis. Improved understanding about the time course and dose response of radiation-induced toxicity will permit more accurate presentation of side effect risk at the time patient consent is obtained. PMID- 16310616 TI - Pain and use of complementary and alternative medicine in a national sample of persons living with HIV. AB - The current study investigated the relationship of pain to use of complementary and alternative medicine (CAM) in a U.S. nationally representative sample of 2466 persons with human immunodeficiency virus (HIV), using data from the HIV Cost and Services Utilization Study. Pain was conceptualized as a need characteristic within the context of predisposing, enabling, and need (PEN) characteristics following Andersen's Behavioral Model of Health Services Use. Multivariate analyses were used to examine the association of baseline PEN characteristics with CAM use by follow-up (approximately 6 months later), including use of five specific CAM domains. Change in pain from baseline to follow-up was also examined in relation to CAM use. Baseline pain was a strong predictor of CAM use, and increased pain over time was associated with use of unlicensed or underground drugs with potential for harm. These results highlight the importance of medical efforts to control pain in persons living with HIV. PMID- 16310618 TI - Cancer-related symptom assessment in Russia: validation and utility of the Russian M. D. Anderson Symptom Inventory. AB - This multicenter cross-sectional study (n=226) validated the Russian-language M. D. Anderson Symptom Inventory (MDASI-R) in Russian cancer patients with hematological malignancies or solid tumors. The Russian-language Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36-R) also was used for validation. Factor analysis found three underlying constructs for symptom items--general, treatment-related, and affective symptoms--with Cronbach alphas of 0.86, 0.68, and 0.90, respectively. Convergent validity was established by comparing MDASI-R items with SF-36-R subscales. The MDASI-R detected significant differences in symptom severity and interference levels by performance status, supporting known group validity. The most prevalent symptoms were fatigue, sleep disturbance, pain, sadness, and poor appetite; 53% of the sample reported one to four moderate to-severe symptoms (>or=5 on 0-10 scale). Symptoms interfered most with work and general activity. Medical professionals underestimated the severity of pain, fatigue, and distress. The MDASI-R is valid and reliable for measuring symptom severity and interference in Russian cancer patients. PMID- 16310619 TI - Cancer pain: voices of the Ojibway people. AB - Recognition of the integration of sensory dimensions of pain with cultural factors has the potential to improve the quality of the clinical encounter between those in pain and their health care providers. This qualitative study used a grounded theory approach to describe the conceptualization of cancer pain from 18 Ojibway patients, family caregivers, and healers/medicine men from a Reserve community and 13 health professionals (nurses and physicians) from a hospital adjacent to this Reserve. Results revealed that for Ojibway participants, cancer pain epitomized all that was most painful in life. Unlike health professionals, they described the properties of pain as a seamless intertwining of the dimensions of physical sensation, threatening cognitions, emotional, social and spiritual anguish, and intuitive sensing. These Ojibway voices on cancer pain remind us of the interrelatedness of culture in the construction of pain and caution us to broaden the restricted focus we bring to the clinical encounter. PMID- 16310620 TI - The impact of chronic pain on depression, sleep, and the desire to withdraw from dialysis in hemodialysis patients. AB - Pain is a multidimensional phenomenon with physical, psychological, and social components and is a significant problem for 50% of hemodialysis (HD) patients. Failure to treat pain adequately may lead to disruption in many aspects of life. This study examines the relationship between moderate to severe chronic pain and depression, insomnia, and the desire to withdraw from dialysis in HD patients. In a cross-sectional study of 205 Canadian HD patients, patients completed a questionnaire that included the Brief Pain Inventory, Beck Depression Inventory, and the Pittsburgh Sleep Quality Index. One hundred and three patients (50.2%) reported chronic pain and 85 (41.4%), moderate to severe pain. There was a higher prevalence of depression in patients with moderate or severe chronic pain compared to patients with mild or no pain (34.1% vs. 18.3%, odds ratio [OR]=2.31, P=0.01). Severe irritability, anxiousness, and inability to cope with stress were all more common in patients with pain compared to patients without pain (P<0.001). There was a higher prevalence of insomnia in patients with moderate or severe chronic pain compared to patients with mild or no pain (74.7% vs. 53.0%, OR=2.32, P=0.02). Although consideration of withdrawal from dialysis was significantly associated with moderate or severe pain compared to no or mild pain (46% vs. 16.7%, P<0.001), death due to withdrawal from dialysis was not. Chronic pain in HD patients is associated with depression and insomnia and may predispose patients to consider withdrawal of dialysis. PMID- 16310621 TI - Psychological functioning associated with prostate cancer: cross-sectional comparison of patients treated with radiotherapy, brachytherapy, or surgery. AB - This study compared the prevalence of psychological difficulties (i.e., anxiety and depression), psychophysiological problems (i.e., insomnia and fatigue), and sexual difficulties across three modalities of treatment for prostate cancer (radiotherapy, brachytherapy, and radical prostatectomy). A total of 861 men completed a battery of questionnaires assessing anxiety, depression, fatigue, insomnia, and quality of life. Patients who initially received radiotherapy had higher levels of depression, anxiety, and fatigue and a lower quality of life, and were more likely to report clinical levels of depression and fatigue. Patients who initially received surgery were more likely to report clinical levels of sexual difficulties but less likely to report clinical levels of depression and fatigue, while patients who received brachytherapy were less likely to report sexual difficulties. Although cross-sectional, these findings raise the possibility of a differential influence of treatments for prostate cancer on some aspects of psychological functioning. PMID- 16310622 TI - Alternative treatments of breakthrough pain in patients receiving spinal analgesics for cancer pain. AB - Patients who experience a poor response to different systemic opioid trials (oral and intravenous) are candidates for spinal treatment. Breakthrough pain occurring in this group of patients is challenging for physicians. This phenomenon has never been described in this context and the treatment is quite difficult, as patients already demonstrated a poor response to systemic opioids. We report a preliminary experience of alternative methods, including the intrathecal injection of local anesthetic boluses as needed, or alternatively, the use of sublingual ketamine. Twelve consecutive patients with advanced cancer and pain were selected for intrathecal treatment after receiving different trials with systemic opioids. During intrathecal therapy, pain flares not responding to high doses of intravenous morphine were treated with intrathecal boluses of local anesthetics titrated to achieve the best balance between analgesia and adverse effects, or with sublingual ketamine (25 mg), according to their preference. Pain and symptoms were recorded for each episode of breakthrough pain during hospital admission. Effective pain control was achieved in all the episodes treated within 10 minutes with either method, without relevant complications. A mean volume of 0.6 mL of levobupivacaine (LB) 0.25% (1.5 mg) was effective within a few minutes and was well tolerated in patients receiving a continuous intrathecal infusion of a combination of morphine and LB in different doses. Similarly, ketamine in doses of 25 mg sublingually was effective and relatively well tolerated. Despite the difficult clinical situation of these patients, these approaches controlled almost all breakthrough pain events previously unresponsive to relatively high doses of intravenous opioids. These intensive treatments should be reserved for a very selected population and initiated in an appropriate setting with frequent monitoring facilities and skilled nursing. PMID- 16310625 TI - Sarcopenia of male aging. PMID- 16310626 TI - Utility of ultrasensitive growth hormone assays in assessing aging-related hyposomatotropism. PMID- 16310627 TI - Aging somatotropic axis: mechanisms and implications of insulin-like growth factor-related binding protein adaptation. PMID- 16310628 TI - Sex-steroid control of the aging somatotropic axis. PMID- 16310629 TI - Testing pituitary function in aging individuals. PMID- 16310630 TI - Perimenopausal reproductive endocrinology. PMID- 16310631 TI - Mechanisms of premature menopause. PMID- 16310632 TI - Mechanisms of hypoandrogenemia in healthy aging men. PMID- 16310633 TI - Relative testosterone deficiency in older men: clinical definition and presentation. PMID- 16310634 TI - Thyrotropin-axis adaptation in aging and chronic disease. PMID- 16310635 TI - Aging-related adaptations in the corticotropic axis: modulation by gender. PMID- 16310636 TI - Pathophysiology of age-related bone loss and osteoporosis. PMID- 16310637 TI - Abnormalities of water homeostasis in aging. PMID- 16310640 TI - Adrenal disorders in pregnancy. PMID- 16310641 TI - Calcium and bone disorders during pregnancy and lactation. PMID- 16310642 TI - Management of gestational diabetes: pharmacologic treatment options and glycemic control. PMID- 16310643 TI - Successful pregnancy in women with type 1 diabetes: from preconception through postpartum care. PMID- 16310644 TI - Pituitary disorders during pregnancy. PMID- 16310645 TI - Thyroid disorders during pregnancy. PMID- 16310646 TI - Fertility in polycystic ovary syndrome. PMID- 16310647 TI - Hypertension in pregnancy. PMID- 16310648 TI - Endocrinology of parturition. PMID- 16310649 TI - Developmental origins of adult metabolic disease. PMID- 16310652 TI - Pediatric anesthesia historical perspective. AB - This article examines how anesthesia evolved to serve the needs of children. Discussion includes milestones in technologic advancement related to pediatric anesthetic care and how collaboration among pediatric surgeons, neonatologists, and pediatric anesthesiologists has helped our specialty to progress. Conversely, the significant contributions of pediatric anesthesiology to pediatric critical care medicine, pain management, and pediatric public health care are also presented. PMID- 16310653 TI - Preoperative psychological preparation of the child for surgery: an update. AB - Preoperative anxiety is associated with a number of poor postoperative outcomes and with significant parental and child distress before surgery. Preparing children for surgery can prevent many behavioral and physiologic manifestations of anxiety. Psychologic and behavioral interventions and pharmacologic interventions are available to treat preoperative anxiety in children. This article discusses the psychologic preparation of children for surgery. PMID- 16310654 TI - The genetics of malignant hyperthermia. AB - The genetic evaluation of the ryanodine type one receptor (RYR1) gene is unlikely to be a useful screening test of malignant hyperthermia susceptibility. But when significant suspicion of malignant hyperthermia has been raised by well documented clinical events or strong family history, the genetic evaluation of RYR1 could secure a diagnosis and indicate appropriate treatment for both the index patient and many relatives of all ages, including the youngest. PMID- 16310655 TI - Opioids in pediatric anesthesia. AB - Opioids are used widely in the practice of pediatric anesthesia and pediatric perioperative medicine. The benefits of opioids are well documented, and their pharmacology has been extensively studied. Nonetheless special care is important when using these agents in the pediatric population. This article addresses the developmental pharmacologic changes that occur with opioids as well as their most common clinical uses. PMID- 16310656 TI - Procedural sedation in the pediatric patient. AB - The demand for safe and effective procedural sedation for children is rapidly increasing because of the increased awareness about procedure-related anxiety even in young infants and children. The development of short-acting sedatives, improved monitoring, and new regulatory requirements have led to the evolution of new paradigms of safe, effective, and resource-efficient systems for providing procedural sedation outside the operating rooms by anesthesiologists and nonanesthesiologists. PMID- 16310657 TI - New developments in pediatric cardiac anesthesia. AB - As the practice of pediatric cardiac anesthesia continues to grow, anesthesiologists now routinely care for patients ranging in size from less than 2 kg to more than 100 kg. New clinical and laboratory research has enhanced our understanding of the effects of anesthetic drugs on the pediatric myocardium, and improvements in survival statistics for even the smallest and sickest infants have shifted the emphasis to evaluation of quality of life and neurological outcome in pediatric cardiac patients. The use of circulatory support in infants and children, both for rapid resuscitation and for more chronic indications such as bridge to transplantation, also continues to evolve, with the recent introduction of pulsatile and axial pumps for pediatric use. This article reviews anesthetic agents, bleeding and coagulation, neurological monitoring, and mechanical circulatory support in the treatment of infants and children. PMID- 16310658 TI - Anesthesia ventilators: better options for children. AB - Mechanical ventilation of pediatric patients in the operating room is challenging. Infants require significantly smaller tidal volumes than adults and changes in delivered volume that would be clinically insignificant for an adult patient, can produce unintended hyper- or hypoventilation in children. The consequences of these unintended ventilation changes can produce hypoxemia, hypercarbia, or barotrauma. This article discusses unique aspects of pediatric ventilation in the operating room, limitations of traditional anesthesia machine technology, the features of modern anesthesia ventilators that circumvent these limitations, and presents several comparison studies. PMID- 16310659 TI - Single-lung ventilation in pediatric anesthesia. AB - Single-lung ventilation is requested for an increasing spectrum of surgical procedures in infants and children. A clear understanding of the physiology of single-lung ventilation, the techniques of lung separation, and the technical skill necessary to apply these techniques are essential for an anesthesiologist practicing thoracic anesthesia. This article focuses on various devices available for single-lung ventilation in the pediatric age group, the relevant respiratory physiology, and the strategies that optimize oxygenation during one-lung anesthesia. PMID- 16310660 TI - Anesthesia considerations for pediatric thoracic solid organ transplant. AB - This article discusses the indications, perioperative management, postoperative complications, and patient outcome of pediatric heart transplantation and pediatric lung transplantation. Special emphasis is placed on the anesthetic considerations relevant for children who are undergoing or have received a solid thoracic organ transplant. PMID- 16310661 TI - Surgery and anesthesia for children who have cerebral palsy. AB - Cerebral palsy is a spectrum of signs and symptoms resulting from an injury to the brain during its developmental stage. Children with cerebral palsy have disorders that impact their anesthetic management. In particular, abnormalities of the respiratory system, gastrointestinal system, and neuromuscular system must be considered. There is emerging evidence for abnormalities in their neuromuscular junction that potentially increase their anesthetic risk. In addition, there are recent data explaining possible reasons for increased bleeding in these children. PMID- 16310662 TI - Anesthesia for pediatric obesity. AB - This article discusses the unique anesthetic implications of obesity, with an emphasis on children and adolescents. It also touches on the issues surrounding bariatric surgery in the morbidly obese adolescent population. Adolescent bariatric surgery is moving to the forefront as a treatment modality because weight-loss programs alone are not keeping pace with the growth of the problem. Bariatric surgery offers the potential to achieve the weight reductions necessary to reverse the debilitating and costly comorbidities of obesity. PMID- 16310663 TI - Anesthetic management for pediatric spinal fusion: implications of advances in spinal cord monitoring. AB - Currently, the detection of emerging injury through intraoperative neurologic monitoring is the best way to prevent neurologic injury. This requires a team approach that includes the anesthesiologist, neurophysiologist, and surgeon. The monitoring modalities available for the patient must be considered in planning the anesthetic management. In addition, intraoperative care for the patient requires an ongoing attention to how the anesthetic drugs affect spinal cord monitoring. PMID- 16310664 TI - Pediatric acute pain management. AB - Children are benefiting from the advances made in developmental neurobiology and analgesic pharmacology over the past few decades. Heightened public awareness and increased political pressure from external regulatory agencies are helping to maintain the momentum in improving pediatric pain management. As a result, methods of assessing and managing children's pain are being refined, and new modalities of pain relief are being explored. This review summarizes selected current topics in pediatric acute pain management, with the major emphasis on acute postoperative pain management. PMID- 16310665 TI - Regional anesthesia in children. AB - The future of regional anesthesia in children is to continue to use current techniques, but also to search for ways to make them easier to employ. The potential development of safe local anesthetic agents with much longer durations, will serve to facilitate improvements in the techniques and styles of practice. The advances in minimally invasive surgical techniques do not mean that regional techniques will not be necessary, but will result in an adaptation of techniques. Peripheral nerve blockade and local wound infiltration can still be used and in some instances, may be very appropriate. PMID- 16310666 TI - Palliative care in pediatrics. AB - Palliative care for children is not about dying; rather, it is about helping children and families to live to their fullest and to restore wholeness while facing complex medical conditions. Family centered pediatric palliative care is the art and science of improving quality of life and attending to suffering for children with life-threatening conditions; the basic principles are presented and discussed in this article. PMID- 16310667 TI - Pediatric anesthesiology: thoughts on the future. AB - Pediatric anesthesiology has made a significant contribution to child health and will be necessary for progress in the health sciences and outcomes related to child health in the future. It is likely that the practice of pediatric anesthesiology will remain an interesting and rewarding but demanding profession for the next generations of physicians. Despite this favorable professional profile, stiff competition for resources will come from other segments of the health care community. This article outlines a multidimensional strategy for pediatric anesthesiology to sustain its progress as a profession and contribute to the health of our children. PMID- 16310670 TI - Cervical neoplasia complicating pregnancy. AB - This article reviews the key concepts regarding the counseling and management of cervical dysplasia and invasive cervical cancer diagnosed during pregnancy. Emphasis is placed on balancing the maternal and fetal well-being in collaboration with appropriate multidisciplinary teams. Information regarding appropriate diagnosis strategies and the impact of delay in treatment, subsequent prognosis, and treatment algorithms are discussed. Novel fertility-sparing techniques for cervical cancer and their impact on complications of future pregnancies are also discussed. PMID- 16310671 TI - Breast cancer and pregnancy. AB - This article addresses a challenging diagnostic and treatment dilemma encountered in the care of pregnant women-breast cancer. The treatment of a breast cancer is significantly affected by an ongoing pregnancy and may result in an increased risk for a poor outcome in the mother. The definition, incidence, mechanism, diagnosis, and treatment of breast cancer associated with pregnancy and the normal physiologic and endocrine changes in the breast during pregnancy that contribute to the difficulty encountered by practitioners in diagnosing and treating the disorder are reviewed. The risks associated with pregnancy after breast cancer treatment and the effect of pregnancy on lifetime risk for breast cancer in the general population and for women with mutations in BRCA1 and BRCA2 are also discussed. PMID- 16310672 TI - Malignant melanoma in pregnancy. AB - This article provides a concise overview of issues relating to melanoma and pregnancy, including pregnancy-associated risk and prognosis, and briefly summarizes results from relevant reports that have been published in recent years. The bulk of evidence amassed over the past half century suggests that pregnancy does not significantly affect the risk of developing malignant melanoma. Further, pregnancy does not seem adversely to influence overall survival from the disease. Most studies found no difference in overall survival between pregnant and nonpregnant women with melanoma. Recent reports from large scale, population-based studies support these conclusions. PMID- 16310673 TI - Malignant adnexal masses in pregnancy. AB - Ovarian tumors during pregnancy are very rare; however, a cancer diagnosis causes distress to the couple. Reassurance is paramount, and the first consideration should be given to the safety of the mother. If both mother and fetus can be preserved, treatment to minimize the risks to both should be planned accordingly. It is imperative to care for the patient with a multidisciplinary team that includes a high-risk obstetrician, a gynecologic oncologist, and a medical oncologist specialized in gynecologic cancers. PMID- 16310674 TI - Hematologic malignancies in pregnancy. AB - Hematologic malignancies complicating pregnancy are uncommon, but a delay in diagnosis or treatment can mean the difference between life and death. It is the responsibility of the obstetrician, nurse-midwife, or nurse practitioner to maintain a high index of suspicion when patients present with unexplained lymphadenopathy or protracted constitutional symptoms. Management of these patients requires a multifaceted team from the oncology, pediatrics, and obstetrics services. With most hematologic cancers now requiring multiagent chemotherapy for optimal survival, the patient, her family, and her physicians are often faced with what seems to be a Faustian dilemma. Most infants exposed in utero to multiagent chemotherapy, however, seem to suffer no long-term detrimental consequences. PMID- 16310675 TI - Gestation trophoblastic diseases: management of cases with persistent low human chorionic gonadotropin results. AB - The finding of persistent low levels of human chorionic gonadotropin (hCG) raises concern, regardless of the antecedent history, and often provokes the evaluating physician to embark on further work-up. The USA hCG Reference Service was started in 1998 to aid physicians with these persistent low hCG cases. This article reviews the observations of the USA hCG Reference Service for 134 cases with persistent low hCG results. Examination of these cases provides clear insight for the appropriate management of those presenting with persistent low levels of hCG. Three distinct sources are discussed for persistent low hCG results: quiescence gestational trophoblastic disease, false-positive hCG results, and pituitary hCG. PMID- 16310676 TI - Chemotherapeutic drugs in pregnancy. AB - Chemotherapy may be indicated for the treatment of cancer during pregnancy. The decision to use chemotherapy significantly impacts the pregnancy, and in turn the pregnancy may affect the treatment options available to patients with cancer. This review provides information about the effects of chemotherapeutic agents in pregnancy, taking into account both the mother and the fetus. For convenience, the agents are divided into categories based upon class and mechanism of action. These include alkylating agents, antimetabolites, nucleoside analogs, topoisomerase I inhibitors, topisomerase II inhibitors, vinca alkaloids, taxanes, and biologics such as signaling and growth factor blocking agents. PMID- 16310677 TI - Gastrointestinal, pancreatic, and hepatic cancer during pregnancy. AB - Pregnancy affects the clinical presentation, evaluation, treatment, and prognosis of patients with gastrointestinal cancer. Pregnant patients may present with advanced gastrointestinal cancer as a result of delayed diagnosis, in part because of difficulty differentiating signs and symptoms of cancer from signs and symptoms of normal pregnancy. The approach to cancer surgery and chemotherapy must be modified in pregnant patients to minimize fetal and maternal risks. Because of these factors, women who develop gastrointestinal cancers during pregnancy seem to have a poor prognosis. This article focuses on cancers of the colon, stomach, pancreas, and liver that occur during pregnancy. PMID- 16310678 TI - Choriocarcinoma and gestational trophoblastic disease. AB - Gestational trophoblastic disease (GTD) encompasses a unique group of uncommon but interrelated conditions derived from placental trophoblasts. For the purposes of discussion GTD is the appropriate collective name for hydatidiform mole, whereas the term gestational trophoblastic neoplasia (GTN) is reserved for cases with persistent human chorionic gonadotropin (hCG) titer elevation after evacuation of hydatidiform mole, metastatic disease, or choriocarcinoma. Although the pathology and clinical behavior of CM and PM are different, the initial management of both conditions is surgical evacuation by suction curettage, determination of the baseline, and follow-up with (hCG) titers. There are guidelines for risk-factor scoring and a staging system that classifies untreated patients into distinct prognostic categories so that treatment outcomes can be objectively compared. The rates of GTN and choriocarcinoma are decreasing and survival has dramatically improved. PMID- 16310679 TI - Validity of the CAGE questionnaire in hospital. AB - OBJECTIVE: The CAGE questionnaire is considered a useful screening and case finding tool for alcohol use disorders in clinical populations. Our objectives were to validate the French version of the CAGE against DSM-IV criteria and to assess performance of each item of the scale. METHOD: Data were extracted from a hospital morbidity study conducted in central France. It concerned 5452 patients 48.5% men-in short and medium-stay units. Patients answered the CAGE questionnaire as a past-year assessment. The alcohol use disorders were diagnosed by the physicians using DSM-IV alcohol abuse or dependency criteria. RESULTS: The CAGE questionnaire for a cut-off of 2 had a sensitivity of 77% and a specificity of 94%. The CAGE test was more sensitive for patients diagnosed as alcohol dependent than for alcohol abusers (61% vs. 84%) with the same specificity (94%). These values are close to those for the English-language CAGE. The first three items (CAG) were very similar, with sensitivity 70% and specificity 94%. The eye opening question (E) differentiated sharply between abuse and dependency, with sensitivities of 18% and 46%, respectively. A questionnaire comprising only the CAG questions of the CAGE had properties similar to the full questionnaire. CONCLUSION: CAGE is a good screening tool for alcohol abuse or alcohol dependency. Given the frequent-and insufficiently diagnosed-alcohol problems among inpatients, CAGE is indicated as a first-line tool for screening for the most severe alcohol use disorders in hospital. It should ideally be used systematically. A positive reply to any of the first three items should alert the clinician and prompt further investigation. PMID- 16310680 TI - Grouping for behavioral and psychological symptoms in dementia: clinical and biological aspects. Consensus paper of the European Alzheimer disease consortium. AB - Behavioral and psychological symptoms of dementia (BPSD), constitute a major clinical component of Alzheimer's disease (AD). There is a growing interest in BPSD as they are responsible for a large share of the suffering of patients and caregivers, and they strongly determine the patient's lifestyle and management. Better detection and understanding of these symptoms is essential to provide appropriate management. This article is a consensus produced by the behavioral group of the European Alzheimer's Disease Consortium (EADC). The aim of this article is to present clinical description and biological correlates of the major behavioral and psychological symptomatology in AD. BPSD is not a unitary concept. Instead, it should be divided into several symptoms or more likely: groups of symptoms, each possibly reflecting a different prevalence, course over time, biological correlate and psychosocial determinants. There is some clinical evidence for clusters within groups of BPSD. Biological studies indicate that patients with AD and BPSD are associated with variations in the pathological features (atrophy, brain perfusion/metabolism, histopathology) when compared to people with AD without BPSD. An individually tailored approach taking all these aspects into account is warranted as it may offer more, and better, pharmacological and non-pharmacological treatment opportunities. PMID- 16310681 TI - Assessing alexithymia in French-speaking samples: psychometric properties of the Observer Alexithymia Scale-French translation. AB - In the present study, we evaluated the psychometric properties of the Observer Alexithymia Scale-French translation (OAS-F), a 33-item, observer-rated alexithymia measure. The scale, accessible to lay and professional raters, taps everyday expressions of alexithymia. French university students (N = 159) were asked to rate a person they knew well or ask an acquaintance to rate them. Those being rated (N = 159) were parents, siblings, children, and friends. OAS-F total and subscale scores were comparable to those in the English normative samples. Moreover, OAS scores were reliable, and the scale's five-factor structure (distant, uninsightful, somatizing, humorless, and rigid) was confirmed. Importantly, too, OAS total scores correlated 0.31 with (self-report) 20-item Toronto Alexithymia Scale (TAS) scores. The OAS-F appears to be a psychometrically sound observer-rated alexithymia measure. PMID- 16310682 TI - Reliability of the Spanish version of the Composite Scale of Morningness. AB - AIM: The aim of this study was to examine the reliability of the Spanish version of Composite Scale of Morningness (CSM) and its ability to measure the circadian typology. SUBJECTS AND METHODS: Voluntary and unpaid psychology students (N= 391; 132 men and 259 women), aged between 17 and 33, completed the questionnaire between the months of September and December. RESULTS: The total score was independent of age and gender, with a close to normal distribution and a non significant negative skewness. The internal consistency was high (Cronbach's alpha= 0.87) and factor analysis extracted three factors labeled Time of Retiring (items 2 and 7), Activity Planning (items 8, 9, and 13) and Morning Affect (items 3-6, and 10-12). With the 10th and 90th percentiles as cut-off scores, scorers below 22 (N= 40; 10.2%) are classified as evening-types and scorers above 39 as morning-types (N= 28; 7.2%). CONCLUSION: The Spanish questionnaire shares most of the good psychometric properties of other versions of the CSM, and thus can be used for Spanish-speaking student samples. Nevertheless, further studies of normative data in workers and aged subjects are needed in order to validate CSM. PMID- 16310684 TI - Intussusception then and now: a historical vignette. PMID- 16310685 TI - Use of resterilized polypropylene mesh in inguinal hernia repair: a prospective, randomized study. AB - BACKGROUND: Inguinal hernia repair consumes considerable health-care resources worldwide. Open mesh repairs are commonly used and the feasibility of using a resterilized mesh, which is a general practice in certain countries, has not been evaluated. STUDY DESIGN: In this randomized prospective study, original and resterilized meshes were used in two groups of patients with unilateral inguinal hernia. Microbiologic changes, textile mechanical properties, overall complication rates, and cost-effectiveness of resterilized mesh were investigated. A time period of 3 years was determined for patient enrollment to this pilot feasibility study, with the goal of 100 patients in each group. RESULTS: Ninety-one patients were enrolled in the original group and 93 in the resterilized mesh group. Median followup was 735 and 739 days and calculated interquartile ranges were 454 and 513 days, respectively. Average tensile strength of the original polypropylene mesh changed slightly with resterilization, as maximum load decreased from a mean of 66.6 to 58.2 N/cm. Overall complication rates were similar in the two groups. The 6.6% infection rate in the original mesh group was not statistically different from the 7.5% rate in the resterilized group (p = 0.80, relative risk = 0.88, 95% confidence interval, 0.31-2.51). There was only one recurrence in the original mesh group in the 21st month. Use of a resterilized mesh decreased the overall cost of operation by decreasing the cost of mesh from 15.9% to 8.3% of the total amount. CONCLUSIONS: Use of a resterilized mesh for inguinal hernia repair is feasible without considerable changes in infection and recurrence rates. PMID- 16310686 TI - The missing thyroid. AB - BACKGROUND: Several patients have been referred to our practice after having undergone attempted thyroidectomy, during which the thyroid gland was not found. These patients were reviewed to identify factors associated with failure to identify or adequately resect the thyroid. STUDY DESIGN: Patients referred for a "missed thyroid" between October 1, 1990 and December 31, 2002, were reviewed retrospectively. Seven patients who underwent neck exploration for thyroid cancer (n=6) or Graves' disease (n=1) were identified. Surgical indications, history of neck operations, means of recognizing the complication, incision location, and intraoperative and pathologic findings were analyzed. RESULTS: The thyroid was missed on initial exploration in four patients and in the second or third operation in three patients. In three patients, the surgeon misidentified the thymus (two patients) or nodal tissue (one patient) as the thyroid gland. In one patient, the thyroid could not be identified. In one patient, the thyroid was deemed unresectable because of scar tissue. In two patients, an inadequate "near total" thyroidectomy was performed. In five patients with long necks, the neck incision was too far below the cricoid cartilage. CONCLUSIONS: Failure to identify the thyroid gland at the normal anatomic position was associated with low cervical incisions and a history of earlier neck operations. These findings emphasize the need for improved understanding of thyroid anatomy, embryology, localization procedures, and surgical technique. PMID- 16310687 TI - Influence of time on risk of bowel resection in complete small bowel obstruction. AB - BACKGROUND: Little is known about the effect of passing time on risk of resection among patients with complete small bowel obstruction. We sought to provide a benchmark of the relationship of time from symptom onset to surgical treatment on the need for resection in patients with complete small bowel obstruction. STUDY DESIGN: We performed an observational study of patients with surgically treated complete small bowel obstruction at an inner-city urban tertiary referral center and a municipal hospital. Patients were sampled randomly retrospectively (n=60), and prospectively (n=81), for a final sample of 141. Detailed clinical and time data were abstracted from medical records including out-of-hospital examinations. Risk of resection was calculated using actuarial life table methods. Linear regression was used to determine factors affecting time to treatment. RESULTS: All patients were treated surgically for obstruction; 45% underwent resection. Resected patients had longer (11 days versus 8 days; p=0.01) and more complicated (31% versus 14% in ICU; p=0.01) hospital stays. The risk of resection was 4% among patients with 24 hours of unresponsive symptoms; it increased to 10% to 14% through 96 hours, then dropped slightly but did not disappear. Patients treated first with a tube had longer times between first examination and operation, system-time (40.6 hours versus 10.2 hours; p=0.0007), but this was not associated with an increased resection risk. System-times were shorter among patients seen first in the emergency department (median: 25.7 hours versus 59.7 hours; p=0.0001). CONCLUSIONS: Physicians should be cautious in postponing surgery beyond 24 hours in patients with unresponsive symptoms from complete obstruction. The risk of resection rises dramatically, remains elevated through 96 hours of unresolved symptoms, then declines but does not disappear. PMID- 16310688 TI - Minimizing local recurrence after breast conserving therapy using intraoperative shaved margins to determine pathologic tumor clearance. AB - BACKGROUND: This study analyzed survival, locoregional recurrence, and reexcision rates after breast conserving therapy, based on the margin analysis technique used at the University of Florida, which incorporates frozen section analysis of shaved breast tissue from the lumpectomy cavity. STUDY DESIGN: A retrospective review was done of 257 patients who underwent 267 consecutive lumpectomy operations and completed radiation therapy at our institution. Margins in 189 patients were assessed using frozen section analysis of shaved margins from the lumpectomy cavity. RESULTS: Breast conserving therapy was performed for 220 (83%) patients with early breast cancer (T1 and T2 tumors) and 47 (17%) with ductal carcinoma in situ. With a median followup of 5.6 years, the crude locoregional recurrence rates for patients who had margins analyzed intraoperatively by frozen section analysis or margins analyzed by permanent analysis were 1.9% (3 of 157) and 3.1% (2 of 63), respectively, for early breast cancer and 15.6% (5 of 32) and 6.6% (1 of 15) for ductal carcinoma in situ (p=NS). Survival rates were 97% and 78%, at 5 and 10 years, respectively, for the early breast cancer patients, and 98% and 98%, respectively, for ductal carcinoma in situ patients. Permanent intraoperative frozen section analysis reexcision rates were 33.3% (26 of 78) and 5.8% (11 of 189). CONCLUSIONS: Regardless of the technique used for margin analysis, breast conserving therapy led to low locoregional recurrence relative to national figures, pointing to the importance of the technique of radiation therapy at our institution. Reexcision rate was reduced with the use of frozen section analysis. PMID- 16310689 TI - Clinical and pathologic features of proximal biliary strictures masquerading as hilar cholangiocarcinoma. AB - BACKGROUND: Nontraumatic inflammatory hilar strictures are uncommon, but are known to mimic malignancy. This study examines the clinical and pathologic features of benign idiopathic strictures. STUDY DESIGN: Patients without a history of trauma or earlier biliary operation treated for benign strictures were identified. Clinical information was obtained from the medical record and all resected specimens were reexamined. RESULTS: From January 1992 to July 2003, 275 patients with proximal biliary strictures were referred. Among these, 22 patients had a final histologic diagnosis of benign stricture, despite a suspected preoperative diagnosis of malignancy. All 22 patients underwent resection of the extrahepatic biliary tree, which in 10 patients was combined with en bloc partial hepatectomy. Histologic reexamination identified five different benign processes: lymphoplasmacytic sclerosing pancreatitis and cholangitis, primary sclerosing cholangitis, granulomatous disease, nonspecific fibrosis/inflammation, and stone disease. Major postoperative morbidity occurred in 6 (26%) patients but none died. No preoperative clinical or radiographic features were identified that could reliably distinguish patients with benign strictures from those with cancer. CONCLUSIONS: "Malignant masquerade" of the proximal bile duct results from several different underlying conditions, and differentiating benign strictures from cancer remains problematic. The treatment approach should continue to be resection for presumed malignancy. PMID- 16310690 TI - Developing quality indicators for elderly patients undergoing abdominal operations. AB - BACKGROUND: Although the expanding and aging population will likely increase demand for surgical services, surgeons and other providers must develop strategies to optimize care. We sought to develop process-based quality indicators for elderly patients undergoing abdominal operations to identify necessary and meaningful ways to improve care in this cohort. STUDY DESIGN: Through structured interviews with thought leaders and systematic reviews of the literature, we identified candidate quality indicators addressing perioperative care in elderly patients undergoing abdominal operations. Using a modification of the RAND/UCLA Appropriateness Methodology, an expert panel of physicians in surgery, geriatrics, anesthesia, internal, and rehabilitation medicine formally rated and discussed the indicators. RESULTS: Eighty-nine candidate indicators were identified and categorized into seven domains: comorbidity assessment (eg, cardiopulmonary disease), elderly issues (eg, cognition), medication use (eg, polypharmacy), patient-to-provider discussions (eg, life-sustaining preferences), intraoperative care (eg, preventing hypothermia), postoperative management (eg, preventing delirium), and discharge planning (eg, home health care). Of the 89 candidate indicators, 76 were rated as valid by the expert panel. Importantly, the majority of indicators rated as valid address processes of care not routinely performed in younger surgical populations. CONCLUSIONS: Attention to the quality of surgical care in elderly patients is of great importance because of the increasing numbers of elderly undergoing operations. This project used a validated methodology to identify and rate process measures to achieve high quality perioperative care for elderly surgical patients. This is the first time quality indicators have been developed in this regard. PMID- 16310691 TI - Developing an appropriate staging system for esophageal carcinoma. AB - BACKGROUND: Development of an optimal staging system for esophageal cancer is essential to estimate prognostic factors accurately and treat them appropriately. In this study, we evaluated the surgical outcomes of esophageal cancer according to five existing staging systems and assessed their prognostic significance. STUDY DESIGN: For 113 patients with esophageal cancer who had undergone curative resection, lymph-node metastasis was classified using the 8th and 9th editions of the Japanese classification, the 6th edition of the Union Internationale Contre le Cancer (UICC) TNM classification, and systems based on the number (0, 1 to 3, or > or = 4) or ratio (0, < 0.15, or > or = 0.15) of metastatic lymph nodes. Survival and prognostic factors of the respective stages were evaluated. RESULTS: Univariate analysis of disease-specific survival revealed that depth of invasion and lymph-node classification notably affected prognosis. Multivariate analysis confirmed that each classification independently influenced prognosis. According to the criteria of the two Japanese classifications, there was no clear correlation between lymph-node stage and survival. The Union Internationale Contre le Cancer/TNM classification, and those based on the number or ratio of metastatic lymph nodes showed a clear correlation between lymph-node metastasis and survival. These systems had better stratification than the Japanese classifications. CONCLUSIONS: Staging systems for esophageal cancer based on the number or ratio of metastatic lymph nodes showed better prognostic significance than those based on the anatomic distribution of metastatic lymph nodes, because of their good stratification and clinical utility. Such classifications are suitable for use throughout the world. PMID- 16310692 TI - Prediction of outcomes in trauma: anatomic or physiologic parameters? AB - BACKGROUND: Prediction of outcomes after injury has traditionally incorporated measures of injury severity, but recent studies suggest that including physiologic and shock measures can improve accuracy of anatomic-based models. A recent single-institution study described a mortality predictive equation [f(x) = 3.48 - .22 (GCS) - .08 (BE) + .08 (Tx) + .05 (ISS) + .04 (Age)], where GSC is Glasgow Coma Score, BE is base excess, Tx is transfusion requirement, and ISS is Injury Severity Score, which had 63% sensitivity, 94% specificity, (receiver operating characteristic [ROC] 0.96), but did not provide comparative data for other models. We have previously documented that the Physiologic Trauma Score, including only physiologic variables (systemic inflammatory response syndrome, Glasgow Coma Score, age) also accurately predicts mortality in trauma. The objective of this study was to compare the predictive abilities of these statistical models in trauma outcomes. METHODS: Area under the ROC curve of sensitivity versus 1-specificity was used to assess predictive ability and measured discrimination of the models. RESULTS: The study cohort consisted of 15,534 trauma patients (80% blunt mechanism) admitted to a Level I trauma center over a 3-year period (mean age 37 +/- 18 years; mean Injury Severity Score 10 +/- 10; mortality 4%). Sensitivity of the new predictive model was 45%, specificity was 96%, ROC was 0.91, validating this new trauma outcomes model in our institution. This was comparable with area under the ROC for Revised Trauma Score (ROC 0.88), Trauma and Injury Severity Score (ROC 0.97), and Physiologic Trauma Score (ROC 0.95), but superior compared with admission Glasgow Coma Score (ROC 0.79), Injury Severity Score (ROC 0.79), and age (ROC 0.60). CONCLUSIONS: The predictive ability of this new model is superior to anatomic-based models such as Injury Severity Score, but comparable with other physiologic-based models such as Revised Trauma Score, Physiologic Trauma Score and Trauma, and Injury Severity Score. PMID- 16310693 TI - Prevention of major amputations in nonischemic lower limb lesions. AB - BACKGROUND: Major lower limb amputations continue to be performed at an increasing rate, the major cause being the rising prevalence of adult onset diabetes. It can be demonstrated that a reduction in amputation rate can be achieved at institutes with a higher level of academic and specialty interest, by avoiding errors of management and by newer technical innovations in wound closure. STUDY DESIGN: A simpler method of wound closure that is easily taught and learned and is readily available to the medical community and the patient population can play a substantial role in reducing the number of amputations. Satisfactory closure of open lesions by full-thickness skin and subcutaneous tissue carries with it a lower incidence of recurrence, reduces costs by avoiding or reducing length of hospitalization, and minimizes use of general anesthesia, a serious matter in immunocompromised older patients. Avoiding errors of technique will additionally reduce amputation rate. RESULTS: In the last 10 years, 125 patients, mostly diabetic and elderly, with major and recurrent lesions of the lower extremities, have been treated by closure of the debrided wound with a combination of external tissue expansion and wound toilette. Major lower limb amputations have been avoided. CONCLUSIONS: Experience confirms that the relationship between high hospital volume, specialty interest, higher level of academic care; avoiding technical errors; and simplifying the methodology, lead to a reduction in need for major amputations. A lower mortality in the patient population at risk is achieved and a reduction in costs, which have risen beyond the 132 billion dollars level, can be expected. PMID- 16310694 TI - PSA testing in office-based clinics: are we testing as much as we think? AB - BACKGROUND: Self-report studies of widespread use of routine prostate-specific antigen (PSA) testing have fueled concerns about overuse and possible harm. The purpose of this study was to examine use of PSA testing during physician office visits in a national sample of prostate-cancer-free men ages 40 years and older. METHODS: Bivariate and multivariate logistic regression models of receiving a PSA test by prostate-cancer-free men ages 40 years and older were performed using the 2000 National Ambulatory Medical Care Survey. RESULTS: There were 2,709 primary care and urology office visits by prostate-cancer-free men 40 years of age and older, and 10.2% resulted in a PSA test. In a multivariate model, men in their 70s had considerably higher odds (odd ratio, 1.60; 95% CI, 1.11-2.32) and men with multiple medical comorbidities had considerably lower odds (odds ratio, 0.28; 95% CI, 0.10-0.65) of receiving a PSA test. CONCLUSIONS: We report lower use of PSA testing than previously published. Greater insight into use of PSA testing based on clinic and hospital administrative data are required to determine the impact of PSA testing on the cost of health care and prostate cancer incidence and mortality. PMID- 16310695 TI - The LANN technique to reduce postoperative functional morbidity in laparoscopic radical pelvic surgery. AB - BACKGROUND: We investigated the feasibility and advantages of introducing Laparoscopic Neuro-Navigation (LANN) into the field of laparoscopic gynecologic radical pelvic surgery. STUDY DESIGN: In a prospective pilot study, 261 consecutive patients underwent laparoscopic radical pelvic surgery for cervical cancer or deep infiltrating endometriosis of the parametria. During the procedure, dissection and electrostimulation, and consequently, sparing of the pelvic parasympathetic nerves by transection of the parametria, were performed. Postoperative bladder dysfunction was documented. RESULTS: Laparoscopic dissection and electrostimulation of the pelvic splanchnic nerves were feasible in all patients without any complications, and the rate of postoperative bladder dysfunction was considerably reduced, to less than 1% of the patients. CONCLUSIONS: The parasympathetic nerve-sparing method using the Laparoscopic Neuro-Navigation technique in laparoscopic radical pelvic gynecologic surgery is a feasible and reproducible technique that preserves postoperative bladder function. PMID- 16310696 TI - Differences in risk factors for lower extremity arterial occlusive disease. AB - BACKGROUND: Patients undergoing lower extremity revascularization have associated cardiovascular risks: smoking, hypertension, dyslipidemia, and diabetes. This study evaluated the impact of cardiovascular risk factors on proximal versus distal arterial occlusive disease in patients undergoing lower extremity revascularization as adjusted to a control group without vascular disease. STUDY DESIGN: We performed a retrospective, case-control study that included 151 patients undergoing lower extremity revascularization and 229 patients undergoing knee and hip replacement (controls). Risk factors were determined for each of three separate groups undergoing revascularization for different levels of occlusive disease: aortoiliac, superficial femoral, and popliteal-tibial. Comparisons to controls were tested using t-tests or chi-square tests and multiple logistic regression. RESULTS: Dyslipidemia was associated with a significant risk of aortoiliac (odds ratio [OR]=3.4; p=0.0006) and superficial femoral occlusion (OR=2.8; p=0.01) but was less strongly associated with popliteal-tibial occlusion (OR=2.1; p=0.09). Smoking was strongly associated with aortoiliac (OR=4.5; p=0.004) and superficial femoral disease (OR=4.6; p=0.0007) but not popliteal-tibial disease (OR=1.3; p=0.53). In contrast, diabetes mellitus and chronic renal insufficiency were strong risk factors for popliteal-tibial occlusion (OR=5.4, p=0.0002; OR=3.9, p=0.01, respectively), but were not significant risk factors for aortoiliac or superficial femoral occlusion. CONCLUSIONS: These data, which use revascularization level as a surrogate marker for lower extremity arterial disease, suggest that the risk factor profile for proximal disease differs from that of distal disease. These findings may reflect differences in the biology of disease and indicate that different risk factors have various anatomic influences on arterial disease formation. PMID- 16310697 TI - Progressive specialization within general surgery: adding to the complexity of workforce planning. AB - BACKGROUND: Although most general surgeons receive comparable training leading to Board certification, the services they provide in practice may be highly variable. Progressive specialization is the voluntary narrowing of scope of practice from the breadth of skills acquired during training; it occurs in response to patient demand, rapid growth of medical knowledge, and personal factors. Progressive specialization is increasingly linked to fellowship training, which generally abruptly narrows a surgeon's scope of practice. This study examines progressive specialization by evaluating trends in fellowship training among general surgeons. STUDY DESIGN: Because no database exists that tracks trainees from medical school matriculation through entrance into the workforce, data from multiple sources were compiled to assess the impact of progressive specialization. Trends in overall number of trainees, match rates, and proportion of international medical graduates were analyzed. RESULTS: The proportion of general surgeons pursuing fellowship training has increased from > 55% to > 70% since 1992. The introduction of fellowship opportunities in newer content areas, such as breast surgery and minimally invasive surgery, accounts for some of the increase. Meanwhile, interest in more traditional subspecialties (ie, thoracic and vascular surgery) is declining. CONCLUSIONS: Progressive specialization confounds workforce projections. Available databases provide only an estimate of the extent of progressive specialization. When surgeons complete fellowships, they narrow the spectrum of services provided. Consequently, as the phenomenon of progressive specialization evolves, a larger surgical workforce will be needed to provide the breadth of services encompassed by the primary components of general surgery. PMID- 16310698 TI - Misdiagnosis of appendicitis and the use of diagnostic imaging. AB - BACKGROUND: CT and ultrasound (US) are increasingly recommended to establish the diagnosis of appendicitis, but population-based rates of misdiagnosis (negative appendectomy [NA]) have not improved over time. The objective of this study was to determine the relationship between CT/US and NA in common practice. STUDY DESIGN: Using data from the Group Health Cooperative on all patients undergoing appendectomy between 1980 and 1999, a longitudinal study was conducted to determine the frequency of NA over time and a case-control, medical record-based study of a subset of patients from the 1990s was conducted to determine the accuracy of CT/US. RESULTS: Of 4,058 patients undergoing appendectomy (mean age 31 +/- 18.6 [SD] years, 49.6% women), 631 (15.5%) had an NA. The overall incidence of NA remained stable over time at 1.5/10,000 patient-years, as did the age and gender adjusted rate (incident rate ratio 0.95, 95% CI 0.97, 1.01). In 1999, nearly 40% of patients had either CT or US. The aggregate sensitivity of these tests was only 74.2% (95% CI 65.7, 83.7), with a positive predictive value of 95.1% (95% CI 91.5, 96.8). CT scans were 88.3% sensitive, with 97.2% positive predictive value (95% CI 92.9, 100). Ultrasounds were 69.5% sensitive, with a positive predictive value of 94.1% (95% CI 89.6, 96.4). More than one in five tests obtained in patients with NA were positive for appendicitis (21.7% for CT and 20.8% for US). CONCLUSIONS: The rate of NA was unchanged over time despite the introduction and use of CT/US, and this appeared to be related to the inconsistent performance characteristics of the tests. This study cautions against overreliance on CT/US in diagnosing appendicitis and emphasizes the need for test benchmarking in routine practice before establishing protocols for presumed appendicitis. PMID- 16310699 TI - Gastrointestinal conditions. PMID- 16310700 TI - The emerging role of 18F-fluorodeoxyglucose positron emission tomography in the management of primary and recurrent rectal cancer. PMID- 16310701 TI - Role of optional (retrievable) IVC filters in surgical patients at risk for venous thromboembolic disease. PMID- 16310702 TI - Compression anastomoses revisited. PMID- 16310703 TI - Medical and surgical care of our four assassinated presidents. PMID- 16310704 TI - Complex aortic atheroma with mobile thrombus. PMID- 16310705 TI - Incarcerated vesicoinguinal hernia presenting with gross hematuria. PMID- 16310706 TI - Management of injuries to the liver and spleen. PMID- 16310707 TI - First civilian helicopter service. PMID- 16310708 TI - Adult necrotizing enterocolitis. PMID- 16310710 TI - Surgical treatment of large contaminated abdominal wall defects. PMID- 16310711 TI - Postinguinal herniorrhaphy pain. PMID- 16310712 TI - Spinal cord stimulation for complex regional pain syndrome: a systematic review of the clinical and cost-effectiveness literature and assessment of prognostic factors. AB - OBJECTIVE: To review the clinical and cost-effectiveness of spinal cord stimulation (SCS) in the management of patients with complex regional pain syndrome (CRPS) and identify the potential predictors of SCS outcome. DESIGN: Systematic review of the literature and meta-regression. METHODS: Electronic databases were searched for controlled and uncontrolled studies and economic evaluations relating to the use of SCS in patients with either CRPS type I or II. RESULTS: One randomised controlled trial, 25 case series and one cost effectiveness study were included. In the randomised controlled trial in type I CRPS patients, SCS therapy lead to a reduction in pain intensity at 24 months of follow-up (mean change in VAS score -2.0), whereas pain was unchanged in the control group (mean change in VAS score 0.0) (p<0.001). In the case series studies, 67% (95% CI 51%, 84%) of type I and type II CRPS patients implanted with SCS reported pain relief of at least 50% over a median follow-up period of 33 months. No statistically significant predictors of pain relief with SCS were observed in multivariate meta-regression analysis across studies. An economic analysis based on the randomised controlled trial showed a lifetime cost saving of approximately 58,470 (60,800 US dollars) with SCS plus physical therapy compared with physical therapy alone. The mean cost per quality-adjusted life year at 12-month follow-up was 22,580 (23,480 US dollars). CONCLUSIONS: SCS appears to be an effective therapy in the management of patients with CRPS type I (Level A evidence) and type II (Level D evidence). Moreover, there is evidence to demonstrate that SCS is a cost-effective treatment for CRPS type I. PMID- 16310713 TI - Can personality traits and gender predict the response to morphine? An experimental cold pain study. AB - The aim of the present study was to examine the possible role of personality traits, in accordance with Cloninger's theory, and gender, in the variability of responsiveness to opioids. Specifically, it was intended to test whether or not the three personality dimensions - harm avoidance (HA), reward dependence (RD) and novelty seeking (NS) - as suggested by Cloninger, can predict inter-personal differences in responsiveness to morphine after exposure to experimental cold pain. Thirty-four healthy volunteers (15 females, 19 males) were given the cold pressor test (CPT). Pain threshold, tolerance, and magnitude (VAS) were measured before and after (six measures, 30 min apart) the administration of either 0.5 mg/kg oral morphine sulphate (n=21) or 0.33 mg/kg oral active placebo (diphenhydramine) (n=13) in a randomized, double blind design. Assessment of the three personality traits, according to Cloninger's Tridimensional Personality Questionnaire, was performed before the CPT. A high HA score (but not RD, NS, or baseline values of the three pain parameters) predicted a significantly larger pain relief following the administration of morphine sulphate (but not of the placebo). Women exhibited a larger response in response to both treatments, as indicated by a significantly increased threshold and tolerance following morphine sulphate as well as significantly increased tolerance and decreased magnitude following placebo administration. The present study confirms the existence of individual differences in response to analgesic treatment. It suggests that high HA personality trait is associated with better responsiveness to morphine treatment, and that females respond better than men to both morphine and placebo. PMID- 16310714 TI - Classification of patients with whiplash associated disorders (WAD): reliable and valid subgroups based on the Multidimensional Pain Inventory (MPI-S). AB - BACKGROUND: Classification of patients with chronic whiplash associated disorders (WAD) into homogenous subgroups is an important objective in order to tailor interventions and to control for subgroup differences when evaluating treatment outcome. AIMS: The aims of this study were to investigate if it was possible to replicate and describe the three cluster solution and profiles found in other pain groups and describe cluster profiles based on self-reported Multidimensional Pain Inventory-scores for patients with WAD three months after the injury, describe characteristics of the clusters in relation to disability, self-efficacy and coping at the same point in time and to validate the cluster solution by comparing clusters in disability, self-efficacy and coping over time. METHODS: Ninety-one WAD-patients three months after the accident took part in the study. The measures used were the Multidimensional Pain Inventory-Swedish version (MPI S), The Self-Efficacy Scale, The Coping Strategies Questionnaire and The Pain Disability Index. Cluster analysis was conducted for the total sample MPI-S subscale scores. RESULTS: The adaptive copers cluster represented 42% of the sample, dysfunctional 34% of the sample, and interpersonally distressed 24% of the sample. The external validation of cluster solution showed that there were several significant differences between clusters in self-efficacy, disability and coping measures. There was also a significant interaction effect (clusterxtime) in disability (PDI). Patients in dysfunctional cluster reported a decreased disability over time. CONCLUSIONS: These results support the presence of different subgroups among patients with whiplash associated disorders. This classification can be seen as a complement to a classification based on medical condition. PMID- 16310715 TI - Secondary heat hyperalgesia induced by melittin in humans. AB - Melittin, which is a principal protein of honeybee venom, can induce mechanical hyperalgesia in humans. The characteristics of the melittin induced mechanical hyperalgesia are quantitatively and qualitatively different from those evoked by capsaicin. The aim of the present study was to investigate in detail secondary heat hyperalgesia induced by melittin in humans. In six healthy volunteers, 10 microg of melittin was injected intradermally on the volar forearm, and VAS score to radiant heat stimuli (focused light from a xenon lamp) was assessed around the injection site 5, 30, and 60 min after injection. For normalization purposes, a pain rating index was calculated as the individual heat evoked VAS scores obtained after melittin divided by the individual baseline VAS scores. A two-way ANOVA revealed a significant increase of the pain rating index over time (F=3.6; P=0.03). The pain rating index at 60 min was significantly larger than at 5 min (P=0.04) and at 30 min (P=0.03). These results demonstrated slowly developing secondary heat hyperalgesia after injection of melittin. A possible contribution of peripheral inflammatory responses to the manifestation of secondary heat hyperalgesia is suggested, which in reality render the distinction between the primary and secondary area of heat hyperalgesia unnecessary. PMID- 16310716 TI - The burden of neuropathic pain: results from a cross-sectional survey. AB - BACKGROUND: There are few published data on the treatment patterns and burden of neuropathic pain. We have investigated this in a large, observational, cross sectional survey. METHODS: We surveyed 602 patients with neuropathic pain recruited from general practitioners in six European countries. Physicians recorded demographic and treatment information, including prescription medications. Patients completed Brief Pain Inventory (BPI) severity and interference questions, the EuroQol (EQ-5D), and questions about their productivity, non-prescription treatments, and frequency of physician visits. The BPI Pain Severity score (range: 0-10) is the mean of worst, least, average, and current pain ratings, with scores of 4-6 and 7-10 considered moderate and severe, respectively. We evaluated the impact of pain severity on functioning using analysis of variance models and chi2 tests. RESULTS: Mean (SD) age was 62.9 (14.4) years (50% female). Most patients reported moderate (54%) or severe (25%) pain. Nearly all patients (93%) were prescribed medications for their neuropathic pain: analgesics (71%); anti-epileptics (51%); antidepressants (29%); sedatives/hypnotics (15%). Seventy-six percent visited their physician at least once in the past month. Employment status was affected in 43% of patients; those employed missed a mean (SD) of 5.5 (9.8) workdays during the past month. Pain severity was associated significantly (P<0.001) with poorer EQ-5D scores (mild=0.67, moderate=0.46, severe=0.16), greater disruption of employment status (mild=24%, moderate=48%, severe=54%), and more frequent physician visits (% with one or more visits: mild=66%, moderate=79%, severe=83%). CONCLUSIONS: Patients with neuropathic pain visit their physician frequently and report substantial pain that interferes with daily functioning despite receiving treatment. PMID- 16310717 TI - Decreased muscle blood flow in fibromyalgia patients during standardised muscle exercise: a contrast media enhanced colour Doppler study. AB - The aim of the study was to investigate if contrast enhanced ultrasound (US) imaging of muscular blood flow during and following exercise could detect alterations in vascularity in fibromyalgia (FM) patients. Ten FM patients and 10 matched controls were examined with US during standardised static and directly following static and dynamic muscular contractions of the infraspinatus muscle. Doppler ultrasound evaluation was performed before and after the administration of ultrasound contrast media. The FM patients had lower magnitude of muscle vascularity following dynamic (p<0.001) and during (p<0.002) static exercise compared to controls. The immediate flow response to muscular activity was not only of a lower magnitude, but also of a shorter duration in FM patients following dynamic exercise (p<0.001) and during static exercise (p<0.01). There were no statistically significant group differences in blood flow intensity or duration following static contraction. In conclusion, contrast enhanced US was found useful to study real-time muscle blood flow changes during and following standardised, low-intensity exercise in FM patients and healthy controls. Our results support the suggestion that muscle ischemia can contribute to pain in FM, possibly by maintaining the central nervous changes such as central sensitisation/disinhibition. US with contrast can be a new valuable approach to assess muscle perfusion in pain patients during standardised exercise. PMID- 16310718 TI - Return to work after comparing outpatient multidisciplinary treatment programs versus treatment in general practice for patients with chronic widespread pain. AB - Former studies have questioned the quality and effectiveness of multidisciplinary rehabilitation for working-age adults with fibromyalgia and chronic widespread pain (CWP). High-quality trials are needed, and return to work should also be included as an outcome variable. This randomized study evaluated the return-to work outcome of an extensive and a light multidisciplinary treatment program combining cognitive strategies and exercise versus treatment-as-usual initiated by a general practitioner, for CWP patients. The patients (n=208), on sick leave for 3 months on average, were randomized to the extensive program including group sessions, a light and more individual program, and to treatment-as-usual. The number of days absent from work and full return to work were used as an outcome, and follow-up lasted 54 months after the programs ended. The regression analysis showed that the extensive program was associated with significantly fewer days absent from work among women. For women, the mean effect of extensive treatment versus treatment-as-usual on total number of days absent from work was estimated to -206.95 days. Among men, the light program was associated with significantly more days absent from work compared to treatment-as-usual. Both among men and women, independent of type of treatment, patients with poorer health (poor prognosis) were absent from work more days than patients with good prognosis. In our sample, higher age significantly increased the number of days absent from work, but only for women. The extensive outpatient multidisciplinary treatment program was effective in returning women to work. PMID- 16310719 TI - Opioid-induced or pain relief-reduced symptoms in advanced cancer patients? AB - BACKGROUND: While opioids in increasing doses may produce adverse effects, the same adverse effects may be associated with poor pain control. Moreover, in the clinical setting symptomatic treatment and illness may balance the outcome of opioid titration. Some adverse effects may tend to disappear continuing the treatment in a long-term period. AIMS: The aim of this study was to monitor the effects of a rapid opioid titration combined with symptomatic treatment in patients with poor relief and to monitor these changes in the following period of 20 days. METHODS: A consecutive sample of 35 patients admitted to an acute Pain Relief and Palliative Care Unit were titrated with opioids, according to a department policy, allowing administration of parenteral opioids to assist opioid titration with oral or transdermal opioids. RESULTS: Thirty-three patients were followed up for the period of the study. Pain was adequately controlled and doses were opioid doses were stable after a mean of 40 h. Opioid escalation index (OEI) was extremely high initially, and then progressively declined at the following study intervals. Weakness and nausea and vomiting did not change, as well as confusion and appetite. Drowsiness, constipation and dry mouth significantly increased and then did not change, although a significant decrease in drowsiness was subsequently observed. Well-being improved some weeks after opioid stabilization. In multivariate analysis, drowsiness and dry mouth were correlated to opioid doses. CONCLUSION: The effects reported were often due to multiple causes. A rapid decrease in pain intensity induced by rapid opioid titration does not produce changes in weakness, nausea and vomiting, appetite. While constipation appears the most relevant problem, resistant to common symptomatic treatment, drowsiness initially produced by acute opioid dose increase and the achievement of pain relief, tends to spontaneously decrease, probably as the result of late tolerance. Improved well-being may be the late positive effect of pain relief, also influenced by the setting of home care. PMID- 16310720 TI - Chronic pain and reduced work effectiveness: the hidden cost to Australian employers. AB - Cost estimates of the impact of chronic pain on work have largely focussed on absenteeism, excluding the hidden contribution of reduced work effectiveness to lost productivity. This paper aims to estimate the cost of lost productivity due to chronic pain in Australia, not only in terms of absenteeism but also reduced work effectiveness. Data on chronic pain in Australia and its impact on work were obtained from the 1997 New South Wales Health Survey (n=17,543) and the Northern Sydney Area Pain Study (n=2,092), both of which were population-based and used random-digit dialing sampling methods. This was combined with relevant Australian population and labour force data obtained from the Australian Bureau of Statistics. From this we estimated annual lost productivity costs arising from both chronic pain-related days absent and 'reduced-effectiveness workdays'. There were estimated to be 9.9 million workdays absent due to chronic pain annually in Australia, equating to a cost of AUD 1.4 billion per annum. Under the assumption that reduced-effectiveness workdays affect productivity costs in the same way as lost work days, the total number of lost workday equivalents was 36.5 million, with the total annual cost of lost productivity due to chronic pain estimated as AUD 5.1 billion per annum. In conclusion, while the impact of reduced work effectiveness on days worked with pain on productivity is uncertain, it has the potential to account for the majority of lost productivity costs associated with chronic pain. Interventions which target working despite pain have the potential to significantly reduce lost productivity costs due to chronic pain. PMID- 16310721 TI - Training medical students to manage a chronic pain patient: both knowledge and communication skills are needed. AB - Most studies concerning pain education of undergraduate medical students focus on knowledge, but little is known about the interviewing skills and pain evaluation. At the end of the fifth study year and at the beginning of the sixth year the students were asked to answer an electronical questionnaire to evaluate how the IASP curriculum on pain had been covered during the studies. In addition, the interviewing skills of the fifth year medical students were assessed using an objective structured clinical examination (OSCE). The students met a standardized patient suffering from postherpetic neuralgia, who was instructed to express depressive and exhausted feelings. A total of 97 students received the questionnaire and 35% responded with identification. All students answering the IASP questionnaire evaluated teaching of postherpetic pain and antidepressant treatment as sufficient. OSCE appeared as a feasible instrument in the assessment of chronic pain education. Eighty-eight percent of the students made the correct diagnosis. However, only 35% asked about sleep disturbances and 16% about depression. When developing a curriculum on pain education, attention should be paid to pedagogic methods about helping the students to implement the learned knowledge in their practice. Formative assessment of both knowledge and skills is essential for the development of a functional pain curriculum. PMID- 16310722 TI - Pulsed radiofrequency applied to dorsal root ganglia causes a selective increase in ATF3 in small neurons. AB - BACKGROUND: This is a "proof of concept study" to test the hypothesis that pulsed radiofrequency, PRF, produces cell stress at the primary afferent level without signs of overt thermal damage. We assumed that cell stress would result in impairment of normal function, and used the expression of activating transcription factor 3, ATF3, as an indicator of cellular "stress". METHODS: PRF (20 ms of 500-kHz RF pulses, delivered at a rate of 2 Hz; maximum temperature 42 degrees C) was delivered either to the sciatic nerve of adult rats in mid thigh, or to the L4 anterior primary ramus just distal to the intervertebral foramen. Controls were sham-operated or L4 axotomised. All tissues were examined 14 days after surgery. The percentage of CGRP- or ATF3-positive DRG neuronal somata was calculated using image analysis software (SigmaScan Pro 4). RESULTS: ATF3 expression was upregulated in L4 DRG neuronal cell bodies, irrespective of their size, after axotomy. It was also upregulated significantly (p<0.002) and selectively, in small and medium calibre L4 DRG neurons, when PRF was applied close to the DRG just distal to the intervertebral foramen. PRF did not produce any obvious cellular changes in the nerve or L4 DRG neurons when applied to the sciatic nerve in mid-thigh. CONCLUSION: PRF has a biological effect, unlikely to be related to overt thermal damage. It appears to be selective in that it targets the group of neurons whose axons are the small diameter C and Adelta nociceptive fibres. PMID- 16310723 TI - Plant metabolomics: potential for practical operation. AB - In the postgenomic era, metabolomics is expected to be the newest useful omics science for functional genomics. However, in plant science, the present metabolomics technology cannot be considered a universal tool to perfectly elucidate perturbations imposed on sample plants although this is desired by plant physiologists. Despite it being an immature technology, metabolomics has already been used as a powerful tool for precise phenotyping, particularly for industrial application. Metabolomics is the best technology for the analysis of large mutant or transgenic libraries of model experimental plants, such as Arabidopsis, rice, etc. Here, we review the applications and technical problems of metabolomics. We also suggest the potential of metabolomics for plant post genomic science. PMID- 16310724 TI - Production of conjugated fatty acids by lactic acid bacteria. AB - Conjugated fatty acids have attracted much attention as a novel type of biologically beneficial functional lipid. Some isomers of conjugated linoleic acid (CLA) reduce carcinogenesis, atherosclerosis, and body fat. Considering the use of CLA for medicinal and nutraceutical purposes, a safe isomer-selective process is required. The introduction of biological reactions for CLA production could be an answer. We screened microbial reactions useful for CLA production, and found several unique reactions in lactic acid bacteria. Lactic acid bacteria produced CLA from linoleic acid. The produced CLA comprised a mixture of cis 9,trans-11-octadecadienoic acid (18:2) and trans-9,trans-11-18:2. Lactobacillus plantarum AKU 1009a was selected as a potential CLA producer. Using washed cells of L. plantarum AKU 1009a as a catalyst, CLA production from linoleic acid reached 40 mg/ml under the optimized conditions. The CLA-producing reaction was found to consist of two successive reactions, i.e., hydration of linoleic acid to 10-hydroxy-12-octadecenoic acid and dehydrating isomerization of the hydroxy fatty acid to CLA. On the basis of these results, the transformation of hydroxy fatty acids by lactic acid bacteria was investigated. Lactic acid bacteria transformed ricinoleic acid (12-hydroxy-cis-9-octadecenoic acid) to CLA (a mixture of cis-9,trans-11-18:2 and trans-9,trans-11-18:2). Castor oil, which is rich in the triacylglycerol form of ricinoleic acid, was also found to act as a substrate for CLA production by lactic acid bacteria with the aid of lipase catalyzed triacylglycerol hydrolysis. L. plantarum AKU 1009a produced conjugated trienoic fatty acids from alpha- and gamma-linolenic acid. The trienoic fatty acids produced from alpha-linolenic acid were identified as cis-9,trans-11,cis-15 octadecatrienoic acid (18:3) and trans-9,trans-11,cis-15-18:3. Those produced from gamma-linolenic were cis-6,cis-9,trans-11-18:3 and cis-6,trans-9,trans-11 18:3. The conjugated trienoic fatty acids produced from alpha- and gamma linolenic acid were further saturated by L. plantarum AKU 1009a to trans-10,cis 15-18:2 and cis-6,trans-10-18:2, respectively. PMID- 16310725 TI - Cytochrome c and bioenergetic hypothetical model for alkaliphilic Bacillus spp. AB - Although a bioenergetic parameter is unfavorable for production of ATP (DeltapH<0), the growth rate and yield of alkaliphilic Bacillus strains are higher than those of neutralophilic Bacillus subtilis. This finding suggests that alkaliphiles possess a unique energy-producing machinery taking advantage of the alkaline environment. Expected bioenergetic parameters for the production of ATP (DeltapH and DeltaPsi) do not reflect the actual parameters for energy production. Certain strains of alkaliphilic Bacillus spp. possess large amounts of cytochrome c when grown at a high pH. The growth rate and yield are higher at pH 10 than at pH 7 in facultative alkaliphiles. These findings suggest that a large amount of cytochrome c at high pHs (e.g., pH 10) may be advantageous for sustaining growth. To date, isolated cytochromes c of alkaliphiles have a very low midpoint redox potential (less than +100 mV) compared with those of neutralophiles (approximately +220 mV). On the other hand, the redox potential of the electron acceptor from cytochrome c, that is, cytochrome c oxidase, seems to be normal (redox potential of cytochrome a=+250 mV). This large difference in midpoint redox potential between cytochrome c and cytochrome a concomitant with the configuration (e.g., a larger negative ion capacity at the inner surface membrane than at the outer surface for the attraction of H+ to the intracellular membrane and a large amount of cyrochrome c) supporting H+-coupled electron transfer of cytochrome c may have an important meaning in the adaptation of alkaliphiles at high pHs. This respiratory system includes a more rapid and efficient H+ and e- flow across the membrane in alkaliphiles than in neutralophiles. PMID- 16310726 TI - Two novel cyclodextrin-degrading enzymes isolated from thermophilic bacteria have similar domain structures but differ in oligomeric state and activity profile. AB - In this paper, we present the expression and characterization of two novel enzymes from the alpha-amylase family exhibiting cyclomaltodextrinase specificity. The nucleotide sequences encoding the enzymes were isolated from the genomic DNA of two thermophilic bacterial strains originating from Icelandic hot springs and belonging to the genera Anoxybacillus (AfCda13) and Laceyella (LsCda13). The genes were amplified using a consensus primer strategy utilizing two of the four conserved regions present in glycoside hydrolase family 13. No identifiable signal peptides were present in open reading frames encoding the enzymes, indicating an intracellular location of both enzymes, and their physiological function to be intracellular cyclodextrin degradation. The domain structures of both enzymes were also similar, including an N-terminal domain, the catalytic module composed of the A- and B-domains, and a C-terminal domain. Despite the similarity in domain composition, the two enzymes displayed differences in the oligomeric state with AfCda13 being a dimeric protein, whereas LsCda13 was monomeric. The two enzymes also displayed significantly different activity profiles, despite being active on the same range of substrates. It was shown that the enzyme displaying the highest activity on cyclodextrin was dimeric (AfCda13). Moreover, a fraction of the dimeric enzyme could be converted to a monomeric state in the presence of KCl and this fraction retained only 23% of its activity on alpha-cyclodextrin while its activity on starch was not significantly affected, indicating that the oligomeric state is an important factor for a high activity on cyclodextrin substrates. PMID- 16310727 TI - Development of simple and efficient in planta transformation method for rice (Oryza sativa L.) using Agrobacterium tumefaciens. AB - Seeds of rice (Oryza sativa L. var. Koshihikari) were soaked in water for 2 d. Thereafter, the embryo containing an apical meristem was inoculated with Agrobacterium tumefaciens by piercing a site of the husk overlying the embryonic apical meristem with a needle that had been dipped in an A. tumefaciens inoculum. The inoculated seeds were then grown to maturation (T0 plants) and allowed to pollinate naturally to set seeds (T1 plants) in pots under nonsterile conditions. To examine the transformation by various means, three different strains of A. tumefaciens were used for transformation: an M-21 mutant, which is an avirulent mutant with a Tn5 insertion in the iaaM gene, and two LBA4404 strains each with a different binary vector. Five different lines of evidence were demonstrated the transformation: the altered phenotype and its inheritance by the next generation, histochemical detection of beta-glucuronidase, resistance to hygromycin B, detection of the transgene by PCR and rescue of a plasmid consisting of the integrated T-DNA and the flanking rice genome DNA. Transformation efficiency of T1 plants was estimated to be 40% and 43% by PCR and a histochemical assay of beta-glucuronidase, respectively. PMID- 16310728 TI - Identification and functional analysis of genes required for desulfurization of alkyl dibenzothiophenes of Mycobacterium sp. G3. AB - Mycobacterium sp. G3 was reported as a dibenzothiophene (DBT)-degrading microorganism and the first strain to have the ability to degrade high-molecular weight alkyl DBTs, such as 4,6-dibutyl DBT and 4,6-dipentyl DBT, by the C-S bond cleavage pathway. Three genes (mdsA, mdsB, and mdsC) for desulfurization, which form a cluster, were cloned from Mycobacterium sp. G3. The expression of each gene in Escherichia coli JM109 showed that MdsC oxidized DBT to DBT sulfone, MdsA transformed DBT sulfone into 2-(2'-hydroxyphenyl)benzene sulfinate (HPBS), and MdsB desulfinated HPBS into 2-hydroxybiphenyl (HBP), indicating that the gene products of mdsABC are functional in the recombinant. MdsC oxidized 4,6-dimethyl DBT, 4,6-diethyl DBT, 4,6-dipropyl DBT and 4,6-dibutyl DBT to each sulfone form, suggesting that MdsC covers a broad specificity for alkyl DBTs. PMID- 16310729 TI - Selection of microalgal growth model for describing specific growth rate-light response using extended information criterion. AB - The effects of light intensity and carbon dioxide (CO2) concentration on the growth of the highly CO2-tolerant green alga Chlorococcum littorale were studied in batch cultures. Four mathematical representations were compared for the specific growth rate-light response curve: a rectangular hyperbolic function, Steele's exponential function, a Poisson function and a hyperbolic tangent function. The hyperbolic tangent function, which is commonly used for representing the photosynthesis-light relationship, gave the best fit as evaluated by the extended information criterion (EIC). EIC proved to be applicable as a criterion to this kind of nonlinear model selection problem. Carbon dioxide, the sole carbon source for photoautotrophic growth of this alga, inhibited the growth rate at concentrations higher than pCO2 of 0.02. A substrate inhibition model was successfully used to simulate the relationship between the specific growth rate and CO2 response. PMID- 16310730 TI - Cloning of alanine racemase genes from Pseudomonas fluorescens strains and oligomerization states of gene products expressed in Escherichia coli. AB - Bacterial alanine racemase (EC 5.1.1.1) is a pyridoxal 5'-phosphate-dependent enzyme. Almost all eubacteria known to date possess a biosynthetic alr gene and some bacteria have an additional catabolic dadX gene. On the basis of the subunit structure, alanine racemases are classified into two types, monomeric and homodimeric. Alanine racemase genes were cloned from two distinct Pseudomonas fluorescens strains, the psychrotrophic TM5-2 strain and the soil-borne LRB3W1 strain, by means of complementing an Escherichia coli alanine racemase-deficient mutant. From the cloning results, both strains are likely to possess only one alanine racemase gene, dadX, in the same manner as the other P. fluorescens strains. Gene organization surrounding the dadX gene is highly conserved among Pseudomonas strains. The gene for D-amino acid dehydrogenase is located adjacent to the dadX gene in both strains. The DadX alanine racemases were expressed in E. coli as C-terminal His-tagged fusion proteins and purified to homogeneity. The catalytic activity of LRB3W1 DadX was higher than that of TM5-2 DadX. The association states of P. fluorescens DadX subunits in the E. coli alanine racemase-deficient mutant were analyzed by gel filtration chromatography. Alanine racemase subunits were demonstrated to exist as both monomers and dimers. The enzyme was in a monomer-dimer equilibrium, and the catalytic activity of the enzyme was proportional to the equilibrium association constant for dimerization. PMID- 16310731 TI - Mesenchymal stem cells differentiate into tenocytes by bone morphogenetic protein (BMP) 12 gene transfer. AB - Rhesus bone marrow mesenchymal stem cells (MSCs) were transfected with the BMP12 gene by electroporation, and the phenotype of the transfected cells was identified by morphological observation and molecular biological assay. After transfection, cells became slender, and their processes became thinner and were interwoven into a network. There were more organelles in the transfected cells than in the parental MSCs. The transfected cells exhibited mRNA expressions of BMP12, collagen type I and scleraxis, but not collagen type III mRNA expression. Immunocytochemical analysis also showed the presence of collagen type I, but not collagen type III in the transfected cells. The transfected cells were positive for CD44 and negative for HLA-DR. Therefore, MSCs can be introduced to differentiate into tenocytes by BMP12 gene transfection, and bone marrow MSCs can serve as an alternative seed cell for application in tendon tissue engineering. PMID- 16310732 TI - Lipase-catalyzed preparation of S-propranolol in presence of hydroxypropyl beta cyclodextrins. AB - A simple method for the preparation of S-propranolol catalyzed by a Rhizopus niveus lipase in an aqueous medium is described. Hydroxypropyl-beta-cyclodextrin was used for the first time to increase the solubility of (R,S)-O-butyryl propranolol thus permitting the reaction to be carried out in water. The formation of an inclusion complex between (R,S)-O-butyryl propranolol and hydroxypropyl-beta-cyclodextrin was studied and a stoichiometry of 1:1 was determined. The influences of the hydroxypropyl-beta-cyclodextrin concentration, pH and percentage of ethanol on the enzymatic activity were also investigated. Under the conditions presented in this paper, values of ee(s) of 90% and E=48 were obtained. PMID- 16310733 TI - Effect of the synthesized mycolic acid on the biodegradation of diesel oil by Gordonia nitida strain LE31. AB - The dynamics of diesel oil biodegradation were previously investigated at initial substrate concentrations of 1000 to 20,000 ppm using Gordonia nitida isolated from wastewater. Following the gas chromatogram profiles of diesel oil degradation, diesel oil with concentrations of up to 15,000 ppm was efficiently degraded by this strain. At a concentrations of 20,000 ppm, however, the degradation by this strain was not effective. The enhancement of the biodegradation of diesel oi1 (at 15,000 and 20,000 ppm) by a synthetic mycolic acid biosurfactant (at 9, 90 and 900 ppm) was also investigated. In G. nitida inoculated cultures, the degradation of diesel oil was enhanced by the biosurfactant. For comparison, diesel oil degradation in batch incubations was measured after the addition of rhamnolipid and other surfactants. Synthetic mycolic acid enhanced the degradation to a greater extent than any other surfactant tested. Additionally, it was demonstrated that the degradation enhancing property of synthetic mycolic acid was similar to that of rhamnolipid and Tween 80. PMID- 16310734 TI - Biological treatment of wastewater discharged from biodiesel fuel production plant with alkali-catalyzed transesterification. AB - The biological treatment of wastewater discharged from a biodiesel fuel (BDF) production plant conducting alkali catalysis transesterification was investigated. BDF wastewater has a high pH and high hexane-extracted oil and low nitrogen concentrations, and inhibits the growth of microorganisms. The biological treatment of BDF wastewater is difficult because the composition of such wastewater is not suitable for microbial growth. To apply the microbiological treatment of BDF wastewater using an oil degradable yeast, Rhodotorula mucilaginosa, the pH was adjusted to 6.8 and several nutrients such as a nitrogen source (ammonium sulfate, ammonium chloride or urea), yeast extract, KH2PO4 and MgSO4.7H2O were added to the wastewater. The optimal initial concentration of yeast extract was 1 g/l and the optimal C/N ratio was between 17 and 68 when using urea as a nitrogen source. A growth inhibitor was also present in the BDF wastewater, and this growth inhibitor could be detected by measuring the solid content in an aqueous phase after the hexane extraction of the wastewater. Microorganisms could not grow at solid contents higher than 2.14 g/l in the wastewater. To avoid the growth inhibition, the BDF wastewater was diluted with the same volume of water. Oil degradation in the diluted BDF wastewater was observed and the best result was obtained under the determined optimal conditions. This treatment system is simple because no controllers, except for a temperature, are necessary. These results suggest that the biological treatment system developed for BDF wastewater is useful for small-scale BDF production plants. PMID- 16310735 TI - Biosynthesis of poly-gamma-glutamic acid in plants: transient expression of poly gamma-glutamate synthetase complex in tobacco leaves. AB - Transient expression of genes coding for the poly-gamma-glutamate (gammaPGA) synthetase system (pgs) was investigated in tobacco plants. Three genes of the pgs, pgsA, pgsB and pgsC, were separately placed under the control of the CaMV 35S promoter and introduced into tobacco leaves via Agrobacterium infection. Synthesized gammaPGA in plant tissues was detected immunologically with mouse anti-gammaPGA antiserum which specifically reacts with gammaPGA on a nitrocellulose membrane. Confirmation of gammaPGA biosynthesis in the transient expression analysis in tobacco tissue indicates that subunits of pgs complex were expressed and reassembled in a functional form. PMID- 16310736 TI - Separation of cultured strawberry cells producing anthocyanins in aqueous two phase system. AB - A rapid and simple selection method of high-yield cells has been desired to establish highly productive cell lines for useful secondary metabolites. For this purpose, a new attempt was made to partition cultured plant cells in a poly(ethylene glycol)-dextran aqueous two-phase system (ATPS). The applicability of the ATPS in partitioning cultured strawberry cells (designated FAW) was investigated. The result of single-step partitioning in the ATPS supplemented with 0.4 mmol/kg lithium sulfate showed that FAW cells cultivated for 7 d under light-irradiation were separated into two cell populations with significantly different anthocyanin content. Additionally, the analysis technique of microscopic cell images showed that cells accumulating a high level of anthocyanin were partitioned completely into the bottom phase in a partitioning experiment of FAW cells cultivated for 10 d under light-irradiation in the ATPS supplemented with 1.8 mmol/kg potassium phosphate buffer. These results indicated that cell partitioning in ATPS increased the intracellular anthocyanin content and that the cultured strawberry cell population was heterogeneous in terms of cell surface properties. This is the first report of partitioning based on the heterogeneity of the cell surface properties correlated with the intracellular secondary metabolism in cultured plant cells. Our results also suggested that the ATPS was appropriate as a large-scale method for selecting useful cell lines among the cultured plant cells. PMID- 16310737 TI - Oxidation of cholesterol catalyzed by amyloid beta-peptide (Abeta)-Cu complex on lipid membrane. AB - A catalytic reaction of H2O2 production by an amyloid beta-peptide (Abeta)-Cu complex with cholesterol incorporated in a liposome was kinetically analyzed. The Michaelis-Menten model was applied to the H2O2 production reaction using cholesterol as the substrate catalyzed by the Abeta-Cu complex. The Km value for the Abeta-Cu complex catalytic reaction with cholesterol-containing 1-palmitoyl-2 oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes (Km=0.436 microM for Abeta(1 40); Km=0.641 microM for Abeta(1-42)) was found to be smaller than that with cholesterol-containing 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) liposomes (Km=0.585 microM for Abeta(1-40), Km=0.890 microM for Abeta(1-42)). The results imply that membrane properties could play an important role in the interactions of the Abeta-Cu complex with cholesterol in these liposomes. Considering the physical states of the cholesterol/POPC (liquid disordered phase) and cholesterol/DPPC (liquid ordered phase) liposomes in the present reaction conditions, the data obtained suggests that the H2O2-generating activity of the Abeta-Cu complex, accompanied by oxidation of membrane-incorporated cholesterol, could be effected by the phase of the liposome membranes. PMID- 16310738 TI - Isolation of copper-tolerant mutants of sake yeast with defective peptide uptake. AB - Mutants of Saccharomyces cerevisiae that have a decreased peptide uptake ability were isolated from sake yeast. A copper medium containing copper sulfate, histidylleucine and sodium lactate as a carbon source was used for the isolation. One mutant showed a decreased peptide uptake ability due to PTR2 transcriptional repression, but for other mutants, the decrease was not due to the transcriptional repression. The peptide concentration in sake brewed with these mutants increased to about 1.5-fold that of sake brewed with the parental strain. The angiotensin I-converting enzyme (ACE) inhibitory activity of sake brewed with the mutant was about 3.6-fold that of sake brewed with the parental strain. PMID- 16310739 TI - Enhanced undecylprodigiosin production from Serratia marcescens SS-1 by medium formulation and amino-acid supplementation. AB - Serratia marcescens Simon Swift-1 (SS-1) was used to produce a prodigiosin-like pigment, undecylprodigiosin (UP), known to have antitumor activities and potential as an anticancer drug. Modified media containing components of Luria Bertani (LB) broth and selected amino acids were used to improve UP production from S. marcescens SS-1. Optimal culture conditions (e.g., temperature, pH, agitation rate) for UP production were also identified. It was found that S. marcescens SS-1 was able to produce 690 mg l-1 of UP when it was grown with 5 g l 1 yeast extract alone (YE medium) under the optimal culture conditions of 30 degrees C, 200 rpm, and pH 8. The UP production of 690 mg l-1 is nearly 23-fold of that obtained from original LB medium. Addition of amino acids containing pyrrole-like structures further enhanced UP production. Nearly 2 and 1.4 g l-1 of UP was produced when the SS-1 strain was cultivated with YE medium supplemented with proline and histidine (5 g l-1), respectively. Moreover, the addition of aspartic acid (5 g l-1) also resulted in a high UP production of 1.4 g l-1. Optimal dosages of the three amino acids were subsequently determined and the highest UP production (2.5 g l-1) was achieved with the addition of 10 g l-1 of proline. This suggests that the supplementation of amino acids related to the formation of a UP precursor (e.g., pyrrolylpyrromethene) could enhance UP production by the SS-1 strain. PMID- 16310740 TI - Cloning and expression of NAD+-dependent L-arabinitol 4-dehydrogenase gene (ladA) of Aspergillus oryzae. AB - A gene of Aspergillus oryzae, ladA, which encodes L-arabinitol 4-dehydrogenase (EC 1.1.1.12), and its cDNA were cloned in Escherichia coli. The gene consisted of a 1209-bp coding region, interrupted by a 59-bp intron, which encoded a 382 amino-acid polypeptide (40,812 Da). The protein showed 67% identity to a well studied L-arabinitol 4-dehydrogenase (Lad1) of Hypocrea jecorina. The cell-free extract of E. coli, which expressed ladA cDNA, showed L-arabinitol dehydrogenase activity with NAD+. It was also reactive for ribitol and xylitol. PMID- 16310741 TI - Enhancement of hepatitis B surface antigen production by adenosine 5' monophosphate in culture of recombinant Chinese hamster ovary cells. AB - Adenosine 5'-monophosphate was found effective in enhancing hepatitis B surface antigen (HBsAg) production in the culture of recombinant Chinese hamster ovary (CHO) cells, but it markedly inhibited cell growth. A culture strategy with periodical medium changes was established to increase HBsAg production and avoid the AMP-induced inhibition of cell growth. The enhanced energy supply might be one of the reasons for the increase in HBsAg production in the presence of AMP. PMID- 16310742 TI - Reaction properties of catalytic antibodies encapsulated in organo substituted SiO2 sol-gel materials. AB - Catalytic antibodies (38C2 and 84G3) were encapsulated in hybrid organic inorganic sol-gel powder prepared from tetraethoxysilane (TEOS) and organoalkoxysilanes, then used to catalyze enantioselective aldol reactions. Sol gel immobilization enhanced antibody stability with respect to the effect of temperature and organic solvent concentration. PMID- 16310743 TI - Trypanosomatid biology and euglenozoan evolution: new insights and shifting paradigms revealed through genome sequencing. PMID- 16310744 TI - The systematic position of the parasitoid marine dinoflagellate Paulsenella vonstoschii (Dinophyceae) inferred from nuclear-encoded small subunit ribosomal DNA. AB - Members of the genus Paulsenella Chatton are marine phagotrophic dinoflagellates that specifically attack marine diatoms. In this phylogenetic study, we show that Paulsenella groups with Amyloodinium ocellatum (Brown) Brown et Hovasse, Pfiesteria piscicida Steidinger et Burkholder (Dinophyceae), Pfiesteria shumwayae Glasgow et Burkholder, and the cryptoperidiniopsoids, all members of the order Peridiniales. In the phylogenetic tree, Paulsenella diverged after Amyloodinium ocellatum but prior to Pfiesteria and the cryptoperidiniopsoids. This suggests that Paulsenella also belongs to the order Peridiniales and its earlier description as gymnodinioid and athecate has to be revised. PMID- 16310745 TI - Molecular evidence for host-symbiont specificity in soritid foraminifera. AB - Symbiosis between the dinoflagellate genus Symbiodinium and various invertebrates and protists is an ubiquitous phenomenon in shallow tropical and subtropical waters. Molecular studies undertaken on cnidarian symbionts revealed the presence of several distinctive lineages or subgeneric clades of Symbiodinium whose taxonomic level provides limited information about the specificity between invertebrate hosts and their symbionts. This contrasts with the finding of several Symbiodinium clades being present almost exclusively in foraminifera and belonging to the subfamily Soritinae. To test whether such specificity also exists at a lower taxonomic level within Soritinae, we obtained the SSU rDNA sequences from 159 soritid individuals collected in nine localities worldwide and representing all known morphospecies of this subfamily. For each individual, the symbionts were determined either by sequencing or by RFLP analysis. We distinguished 22 phylotypes of Soritinae in relation with a number of symbiont "groups" corresponding to 3 clades and 5 subclades of Symbiodinium. Among the 22 soritid phylotypes, 14 show strict symbiont specificity and only one was found to be a host for more than two "groups" of Symbiodinium. It is suggested that the strong host-symbiont specificity observed in Soritinae is a combined effect of a selective recognition mechanism, vertical transmission of symbionts, and biogeographical isolation. PMID- 16310746 TI - Phylogenetic position of the copepod-infesting parasite Syndinium turbo (Dinoflagellata, Syndinea). AB - Sequences were determined for the nuclear-encoded small subunit (SSU) rRNA and 5.8S rRNA genes as well as the internal transcribed spacers ITS1 and ITS2 of the parasitic dinoflagellate genus Syndinium from two different marine copepod hosts. Syndinium developed a multicellular plasmodium inside its host and at maturity free-swimming zoospores were released. Syndinium plasmodia in the copepod Paracalanus parvus produced zoospores of three different morphological types. However, full SSU rDNA sequences for the three morphotypes were 100% identical and also their ITS1-ITS2 sequences were identical except for four base pairs. It was concluded that the three morphotypes belong to a single species that was identified as Syndinium turbo, the type species of the dinoflagellate subdivision Syndinea. The SSU rDNA sequence of another Syndinium species infecting Corycaeus sp. was similar to Syndinium turbo except for three base pairs and the ITS1-ITS2 sequences of the two species differed at 34-35 positions. Phylogenetic analyses placed Syndinium as a sister taxon to the blue crab parasite Hematodinium sp. and both parasites were affiliated with the so-called marine alveolate Group II. This corroborates the hypothesis that marine alveolate Group II is Syndinea. PMID- 16310747 TI - A plastid in the making: evidence for a second primary endosymbiosis. AB - One of the major steps in the evolution of life was the origin of photosynthesis in nucleated cells underpinning the evolution of plants. It is well accepted that this evolutionary process was initiated when a photosynthetic bacterium (a cyanobacterium) was taken up by a colorless host cell, probably more than a billion years ago, and transformed into a photosynthetic organelle (a plastid) during a process known as primary endosymbiosis. Here, we use sequence comparisons and phylogenetic analyses of the prokaryotic rDNA operon to show that the thecate, filose amoeba Paulinella chromatophora Lauterborn obtained its photosynthetic organelles by a similar but more recent process, which involved a different cyanobacterium, indicating that the evolution of photosynthetic organelles from cyanobacteria was not a unique event, as is commonly believed, but may be an ongoing process. PMID- 16310748 TI - The lure of life cycles: Cyril Garnham and the malaria parasites of primates. PMID- 16310749 TI - Driver Behaviour Questionnaire: a follow-up study. AB - The aim of the present study was to investigate time-across stability of different factor solutions (two to six factors) of the Driver Behaviour Questionnaire (DBQ) and to examine the changes on self-reported driving pattern in a follow-up sample (n=622) after three years of the first responses. Repeated measures ANOVA indicated that there was a significant change between Time 1 and Time 2 scores in six items of the DBQ. Drivers reported less competitiveness while driving at Time 2 but more speeding, drinking and driving, driving to wrong destinations and having no recollection of the road just travelled. Significant Time x Sex x Age interactions were found in change scores of four items. Young males and middle-aged female drivers emerged as a group of drivers who changed their self-reported driving pattern over three years. Additionally, sex, age or both had main effects on scores of 21 items. Males and young drivers reported more violations than females and older drivers, whereas female drivers reported more errors and lapses. After running possible factor solutions with Tucker's Phi agreement coefficients, the results indicated that the four- and two-factor solutions were the most stable and interpretable ones. The two-factor solution showed better time-across stability than the four-factor structure did, although the factor solutions found at Time 1 and Time 2 were not as identical as expected. Separate analysis revealed that drivers who had high annual mileage at Time 1 and Time 2 showed the strongest two-factor time-across stability. The test retest reliability was 0.50 for errors, 0.76 for violations and 0.61 for the whole scale. PMID- 16310750 TI - Engrossed in conversation: the impact of cell phones on simulated driving performance. AB - The current study examined the effects of cognitively distracting tasks on various measures of driving performance. Thirty-six college students with a median of 6 years of driving experience completed a driving history questionnaire and four simulated driving scenarios. The distraction tasks consisted of responding to a signal detection task and engaging in a simulated cell phone conversation. Driving performance was measured in terms of four categories of behavior: traffic violations (e.g., speeding, running stop signs), driving maintenance (e.g., standard deviation of lane position), attention lapses (e.g., stops at green lights, failure to visually scan for intersection traffic), and response time (e.g., time to step on brake in response to a pop-up event). Performance was significantly impacted in all four categories when drivers were concurrently talking on a hands-free phone. Performance on the signal detection task was poor and not significantly impacted by the phone task, suggesting that considerably less attention was paid to detecting these peripheral signals. However, the signal detection task did interact with the phone task on measures of average speed, speed variability, attention lapses, and reaction time. The findings lend further empirical support of the dangers of drivers being distracted by cell phone conversations. PMID- 16310751 TI - Is electronic stability program effective on French roads? AB - This paper proposes an evaluation of the effectiveness of the electronic stability program (ESP) in terms of reduction of injury accidents in France. The method consists of 3 steps: The identification, in the French National injury accident census, of accident-involved cars for which the determination of whether or not the car was fitted with ESP is possible. A sampler of 136 cars involved in injury accidents occurred in 2000, 2001, 2002 and 2003 was then selected. But we had to restrict the analysis to only 588 Renalut Laguna's. The identification of accident situations for which we can determine whether or not ESP is pertinent ( for example ESP is pertinent for loss of control accidents whilst it is not for cars pulling out of a junction). The calculation, via a logistic regression, of the relative risk of being involved in an ESP-pertinent accident for ESP equipped cars versus unequipped cars, divided by the relative risk of being involved in a non-ESP-pertinent accident for ESP equipped cars versus unequipped cars. This relative risk is assumed to be the best estimator of ESP effectiveness. The arguments for such a method, effectiveness indicator and implicit hypothesis are presented and discussed in the paper. Based on a few assumptions, ESP is proved to be likely effective. Currently, the relative risk of being involved in an ESP pertinent accident for ESP-equipped cars is lower (-44%, although not statistically significant) than for other cars. PMID- 16310752 TI - On the occurrence of thioredoxin in Trypanosoma cruzi. AB - The full coding sequence for thioredoxin from Trypanosoma cruzi (TcTRX) strain Tulahuen O has been cloned into the pRSETA vector. The protein was expressed in Escherichia coli with an N-terminal extension of six histidine residues for purification through metal ion chromatography. The biological activity of recombinant TcTRX was proved utilizing the insulin reduction assay. Amino acid sequence alignment indicates a high identity of TcTRX with thioredoxins from different sources. Immunocytochemistry assays showed that TcTRX is present in epimastigote forms of T. cruzi, thus, indicating that the gene is expressed in vivo, rather than being a pseudogene. The in vivo occurrence of TcTRX points out the necessity of considering this protein as a molecular component of the redox metabolism in trypanosomatids. PMID- 16310753 TI - Direct genotyping of animal and human isolates of Toxoplasma gondii from Colombia (South America). AB - Genetic analysis of the SAG2 locus was performed to determine the prevalence of the main genotypes of Toxoplasma gondii (SAG2 types I, II, and III) associated with humans, cats, birds and guinea pig toxoplasmosis in Colombia. This typing was directly performed on clinical samples and autopsy material from human or animals. A total of 50 from 146 samples were positive by specific B1 Toxoplasma PCR assay and then were analyzed by restriction fragment length polymorphism in PCR-amplified SAG2 products. Characterization of the SAG2 gene was successful in 33 (66%) of the samples. Genotyping indicated that 31 (93.9%) were SAG2 type I, 1 was SAG2 type III and 1 was atypical. In birds and cats all samples were SAG2 type I. Results support a predominance of the Toxoplasma SAG2 type I circulating in human and animals in South America. PMID- 16310754 TI - An automated peptide and protein thiazolidine coupling chemistry for biosensor immobilization giving a unique N-terminal orientation. PMID- 16310755 TI - Kinetic and thermodynamic analysis of Bradyrhizobium japonicum PutA-membrane associations. AB - In Escherichia coli, proline induces tight membrane binding of the PutA flavoenzyme and transforms PutA from a transcriptional repressor to a membrane associated proline catabolic enzyme. In other gram-negative bacteria such as Bradyrhizobium japonicum, PutA lacks DNA binding activity and functions only as a proline catabolic enzyme. Here, we characterize the membrane binding properties of PutA from B. japonicum (BjPutA) to address whether proline regulates BjPutA lipid binding similar to Escherichia coli PutA (EcPutA). Surface plasmon resonance (SPR) kinetic measurements of BjPutA-lipid binding show BjPutA forms a complex with lipids in the absence and presence of proline with similar dissociation constant (K(D)) values of 2.5 and 1.7nM, respectively. SPR experiments using differently charged lipid bilayers indicate BjPutA selectively binds negatively charged lipids, which contrasts with the charge independent membrane binding of EcPutA. Analysis of BjPutA-lipid binding by isothermal titration calorimetry at 25 degrees C revealed an endothermic binding reaction that is entropically driven. This work shows that BjPutA-membrane associations vary significantly from EcPutA. PMID- 16310756 TI - Histamine (H3) receptors modulate the excitatory amino acid receptor response of the vestibular afferents. AB - Although the effectiveness of histamine-related drugs in the treatment of peripheral and central vestibular disorders may be explained by their action on the vestibular nuclei, it has also been shown that antivertigo effects can take place at the peripheral level. In this work, we examined the actions of H3 histaminergic agonists and antagonists on the afferent neuron electrical discharge in the isolated inner ear of the axolotl. Our results indicate that H3 antagonists such as thioperamide, clobenpropit, and betahistine (BH) decreased the electrical discharge of afferent neurons by interfering with the postsynaptic response to excitatory amino acid agonists. These results lend further support to the idea that the antivertigo action of histamine-related drugs may be caused, at least in part, by a decrease in the sensory input from the vestibular endorgans. The present data show that the inhibitory action of the afferent neurons discharge previously described for BH is not restricted to this molecule but is also shared by other H3 antagonists. PMID- 16310757 TI - Comparison of curdlan and its carboxymethylated derivative by means of Rheology, DSC, and AFM. AB - Curdlan was carboxymethylated in an aqueous alkaline medium using monochloroacetic acid as the etherifying agent. The structure of carboxymethylated curdlan (CMc) was analyzed by FT-IR and NMR spectroscopy, which revealed that the carboxymethyl group was introduced mainly at the C-6 position as well as at the C-2 and C-4 positions. Furthermore, CMc was compared with the native curdlan by using rheology and DSC methods. It was found that in water, both polysaccharides behaved as pseudoplastic fluids and fit the power law and Herschel-Bulkley rheological models well. Both the storage shear modulus G' and the loss shear modulus G'' of CMc aqueous solutions decreased and became more frequency dependent with decreasing concentration in comparison with the curdlan aqueous suspensions. The modulus-temperature curve also suggested that the gel characteristic of curdlan has been lost after chemical modification, which is consistent with the DSC results. AFM images revealed differences in the conformation of native and carboxymethylated curdlan, which changed from the aggregation of macromolecules to triple helices. All the experimental results suggest that the hydrogen bonds that bind curdlan with interstitial water to form the micelles have been destroyed completely and that the hydrophobic interactions related to the methylene groups at C-6 formed above 55 degrees C disappeared due to the introduction of the hydrophilic carboxymethyl group. PMID- 16310758 TI - Behavior of GM3 ganglioside in lipid monolayers mimicking rafts or fluid phase in membranes. AB - We studied the interaction of GM3 ganglioside with sphingomyelin (SM) and palmitoyl-oleoyl-phosphatidylcholine (POPC) in Langmuir monolayers mimicking, respectively, raft and fluid phase of a cellular membrane, by surface pressure measurements and fluorescence microscopy. No difference was observed in the behavior of SM-GM3 and POPC-GM3 monolayers. In both cases, a GM3 threshold concentration has been underlined between 20 and 40 mol%. Below this threshold, SM-GM3 and POPC-GM3 monolayers behave ideally, suggesting that GM3 and host lipid would form separated domains. On the contrary, above the threshold, a condensation of monolayers is observed. This could be due to a partial solubilisation of GM3 in host lipid, leading to a change in orientation of GM3 molecules at the air-water interface. PMID- 16310759 TI - Practical considerations in PTH testing. AB - New knowledge concerning PTH biology have accumulated during the past few years. The finding that the so-called "intact" PTH assays measure a "non-1-84" PTH fragment in addition to full-length PTH has led to the development of new assays. These new assays, which were initially thought to measure 1-84 PTH only, have been shown to recognize also another PTH species called "amino-PTH". As the various names given to the different assay methods are highly confusing, there is a need for a simplified nomenclature. A simple way would be to identify the older "intact" PTH assays as second-generation assays and the new assays (Whole, CAP, BioIntact) as third-generation assays. Although of considerable potential interest for the comprehension of PTH physiology, the third-generation PTH assays have not yet proved to be superior to the second-generation assays in clinical practice. There is thus currently no recommendation to switch from the second generation to the third-generation assays in clinical practice, or to use a ratio derived from the concommitent measurement of PTH with both assay-generation. Because second- and third-generation PTH assays are usually highly correlated, significant differences in the clinical information provided by these methods are unlikely. However, our opinion is that more definitive bone biopsy studies in dialyzed patients selected according to their bone- and calcium-related treatment are still needed to reach a consensus. Finally, we have proposed that PTH reference values should be established in healthy subjects with a normal vitamin D status. This supposes that 25OHD is measured in the reference population beforehand, and that the subjects with vitamin D insufficiency are eliminated from the reference group. Although more complicated than the usual way to establish normative data, we have shown that it decreases the upper limit of normal by 25-35%, enhancing thus the diagnostic sensitivity for hyperparathyroidism without a decrease in specificity. PMID- 16310760 TI - Detection of electronegative low density lipoprotein (LDL-) in plasma and atherosclerotic lesions by monoclonal antibody-based immunoassays. AB - OBJECTIVES: To produce a monoclonal antibody (MAb) against electronegative LDL (LDL-) for detecting this modified lipoprotein in blood plasma and tissues. DESIGN AND METHODS: LDL- was isolated from human blood plasma and used as an antigen for immunization of Balb/c mice. Lymphocytes of immunized mice were fused with myeloma cells (SP2/0) to obtain the hybridomas. LDL- was detected in blood plasma and atherosclerotic lesions of humans and rabbits by MAb-based ELISA and immunohistochemistry, respectively. RESULTS: LDL- concentrations were higher (P < 0.05) in the blood plasma of hypercholesterolemic subjects (HC, 248 +/- 77 mg/dL of total cholesterol) than in normolipidemic subjects (NL, 173 +/- 82 mg/dL of total cholesterol) and rabbits (HC, 250 +/- 15 mg/dL of cholesterol versus NL, 81 +/- 12 mg/dL of cholesterol). Moreover, LDL- was detected in the atherosclerotic lesions of humans and rabbits. CONCLUSION: These MAb-based immunoassays are adequate to detect LDL- in biological samples and represent an important tool for investigating the role of LDL- in atherosclerosis. PMID- 16310761 TI - Processing controlled PROs in Spanish. AB - Two experiments were carried out to investigate the processing of the empty category PRO and the time-course of this in Spanish. Eye movements were recorded while participants read sentences in which a matrix clause was followed by a subordinate infinitival clause, so that the subject or the object of the main clause could act as controller of PRO, and therefore as implicit grammatical subject of the infinitive. In Experiment 1, verb control information was manipulated: The matrix clause contained either subject-control verbs like prometer ('promise') or object-control verbs like forzar ('force'). In Experiment 2, the preposition that headed adverbial subordinate clauses was manipulated: Two different kinds of infinitival adverbial clauses were used, expressing purpose (preposition para) and reason (preposition por) and in which control information is primarily induced by the prepositions (para tends to trigger subject-control; por object-control). Experiment 1 showed that readers make immediate use of verb control information to recover the antecedent of the empty category PRO in Spanish obligatory control constructions. The data obtained in Experiment 2 suggest that during the processing of the empty category PRO in purpose vs. reason adverbial subordinate infinitival clauses the control information induced by the prepositions por vs. para is not initially used as a constraint to guide the selection of the nominal antecedent of PRO. In addition, both experiments showed that PRO antecedent selection is a very fast process and that, together with verb control information, recency played an important role. The results are discussed in terms of current psycholinguistic hypotheses about empty categories and in relation to formal linguistic hypotheses about PRO control. We argue that obligatory control ties (which are co-referential ties) are processed fast because they are launched from the same lexical platform that launches all fast syntactic connections (complements, as opposed to modifiers). In short, that such lexically-specified co-referential ties are regulated by linguistic form. PMID- 16310762 TI - Retinal stem/progenitor properties of iris pigment epithelial cells. AB - Neural stem cells/progenitors that give rise to neurons and glia have been identified in different regions of the brain, including the embryonic retina and ciliary epithelium of the adult eye. Here, we first demonstrate the characterization of neural stem/progenitors in postnatal iris pigment epithelial (IPE) cells. Pure isolated IPE cells could form spheres that contained cells expressing retinal progenitor markers in non-adherent culture. The spheres grew by cell proliferation, as indicated by bromodeoxyuridine incorporation. When attached to laminin, the spheres forming IPE derived cells were able to exhibit neural phenotypes, including retinal-specific neurons. When co-cultured with embryonic retinal cells, or grafted into embryonic retina in vivo, the IPE cells could also display the phenotypes of photoreceptor neurons and Muller glia. Our results suggest that the IPE derived cells have retinal stem/progenitor properties and neurogenic potential without gene transfer, thereby providing a novel potential source for both basic stem cell biology and therapeutic applications for retinal diseases. PMID- 16310763 TI - The bZip proteins CES-2 and ATF-2 alter the timing of transcription for a cell specific target gene in C. elegans. AB - The Caenorhabditis elegans basic leucine zipper (bZip) protein CES-2 regulates the transcription of different genes in different cells. Here, we show that CES-2 mediates its effects in one cell (the excretory duct cell) together with another bZip protein, ATF-2. Both proteins influence transactivation of a target gene, lin-48. Although lin-48 is required for both anatomical and physiological features of the excretory duct cell, ces-2 and atf-2 mutants only exhibit anatomical defects. We show that CES-2 and ATF-2 modify the temporal transcription of lin-48, acting to initiate embryonic expression required for normal excretory system development and anatomy. In contrast, CES-2 and ATF-2 do not influence post-embryonic lin-48 expression. We provide evidence that CES-2 and ATF-2 can interact with each other, and that they cooperate to form a complex on lin-48 regulatory sequences. Thus, the two bZip proteins function together as one of several inputs that influence the spatial and temporal regulation of lin 48. PMID- 16310764 TI - Hydrochlorothiazide increases plasma or tissue angiotensin-converting enzyme inhibitor drug levels in rats with myocardial infarction: differential effects on lisinopril and zofenopril. AB - Sodium depletion with diuretics augments the efficacy of angiotensin-converting enzyme-inhibitor therapy for hypertension and renal dysfunction, and possibly for left ventricular dysfunction after myocardial infarction. Underlying mechanisms may involve altered angiotensin-converting enzyme-inhibitor pharmacokinetics. We hypothesized that the diuretic hydrochlorothiazide causes increased steady-state levels of the angiotensin-converting enzyme-inhibitors lisinopril and zofenopril in rats with myocardial infarction. Rats were subjected to coronary ligation to induce myocardial infarction. After 1 week, rats were randomized to 50 mg/kg/day hydrochlorothiazide or control treatment for 3 weeks. The last week, rats received lisinopril or zofenopril in equipotentent dosages (3.3 and 10 mg/kg/day, respectively). Rats were sacrificed at Tmax after the last dose of angiotensin converting enzyme-inhibitor, and tissues were collected for analysis of drug concentrations. Lisinopril concentrations in plasma were significantly increased by hydrochlorothiazide, at unchanged tissue concentrations. This increase could be fully explained by decreased renal function, as evidenced by increased plasma creatinine levels (lisinopril + hydrochlorothiazide 82+/-5 microM versus lisinopril 61+/-5 microM, P < 0.001). In contrast, zofenoprilat levels in kidney and non-infarcted left ventricle were markedly increased by hydrochlorothiazide, whereas plasma concentrations were unchanged. Although hydrochlorothiazide tended to increase plasma creatinine in zofenopril-treated rats as well, this increase was less pronounced (zofenopril + hydrochlorothiazide 61+/-3 microM versus zofenopril 54+/-2 microM, P = 0.15). Hydrochlorothiazide increases steady-state angiotensin-converting enzyme-inhibitor drug levels, most likely by affecting their renal clearance. Notably, the lipophilic angiotensin-converting enzyme inhibitor zofenopril accumulated in tissue, whereas the hydrophilic lisinopril increased in plasma. Whether combining different angiotensin-converting enzyme inhibitors with hydrochlorothiazide translates into distinct clinical profiles requires further study. PMID- 16310765 TI - The cysteinyl-leukotriene-1 receptor antagonist zafirlukast is a potent secretagogue in rat and human airways. AB - Cysteinyl-leukotriene-1 receptor antagonists are important tools in the therapy of asthma. Although many studies have been performed concerning their effects on airway smooth muscle tone, there are no basic data on their effects on airway secretions. Therefore, we assessed the effects of zafirlukast and montelukast on rat tracheal secretion by quantification of secreted 35S04 labelled mucus macromolecules, and determined the influence of the arachidonic acid pathway using the modified Ussing chamber technique. Zafirlukast (432+/-89.99%) and montelukast (167+/-16.74%) stimulated rat tracheal secretion. This was abolished by application of eicosatetraenoic acid, an inhibitor of the arachidonic acid metabolism. Whereas inhibition of cyclooxygenase did not show any significant effect on zafirlukast induced secretion, blockade of the 5-lipoxygenase pathway markedly reduced the secretagogue effects. Furthermore, inhibition of phosphatidylinositol-3-kinase completely inhibited the effects elicited by zafirlukast. Additional experiments revealed secretagogue effects of zafirlukast also in human bronchial tissue. In conclusion, zafirlukast is a potent inducer of tracheal secretion. Obviously, these effects are induced by involvement of a phosphatidylinositol-3-kinase dependent pathway mediated by products of the arachidonic acid metabolism. PMID- 16310766 TI - Violence among peoples in the light of human frustration and aggression. AB - This article sets out to provide a general background to the study of aggression in the social sciences, with a particular focus on its link to collective violence. While the study of what happens in the human brain appears to be already highly complex, analysis of violent behavior appears to be even more intricate. A deductive system in the sense of a general and clear system of propositions logically connected to one another is not feasible, principally because contrary to the natural sciences there are no verities but merely "stylized facts." One of these concerns the setting of human aggression in the light of frustration, as argued in the frustration-aggression hypothesis developed by Dollard et al. in 1939. Apart from conceiving of aggression as a pure human instinct, it may also be seen as externally driven, while a third possibility concerns culturally "learned" aggression. Proof of the latter is that the strongest correlation appears to be that between current violence and previous manifestations thereof. Attention is paid to the way in which Gurr has rooted his relative deprivation theory on causes of collective violence among peoples in mechanisms of frustration and aggression. That theory is taken a bit further in terms of "perceived acquirement failure," which appears to be highly connected to the role of the state. Based on certain observations by Hannah Arendt, the argument then proceeds to violence as a manifestation of powerlessness. Finally, this leads to a discussion of justice as a crucial factor in what Durkheim used to call a "right to conflict." In this way, human aggression is placed in a broad socio-economic context. PMID- 16310767 TI - Treatment with PARP-1 inhibitors, GPI 15427 or GPI 16539, ameliorates intestinal damage in rat models of colitis and shock. AB - Poly (ADP-ribose) polymerase-1 (PARP-1), a nuclear enzyme activated by DNA strand breaks, plays a detrimental role during inflammation. As inflammation is important in the development of colitis and ischemia/reperfusion (I/R) injury of the intestine, we investigated the effects of 10-(4-methyl-piperazin-1-ylmethyl) 2H-7-oxa-1,2-diaza-benzo[de]anthracen-3-one (GPI 15427) and 2-(4-methyl-piperazin 1-yl)-5H-benzo[c][1,5]naphthyridin-6-one (GPI 16539), two novel and potent inhibitors of PARP-1, in a rat model of gut injury and inflammation, splanchnic artery occlusion (SAO)shock and dinitrobenzene sulfonic acid (DNBS)-induced colitis. We report here for the first time that post-injury administration of GPI 15427 and GPI 16539 exerts potent anti-inflammatory effects by reducing inflammatory cell infiltration and histological injury, and delaying the development of clinical signs in both in vivo models. Furthermore, GPI 15427 and GPI 16539 treatment diminished the accumulation of poly(ADP-ribose) in the ileum of splanchnic artery occlusion-shocked rats and in the colons of dinitrobenzene sulfonic acid-treated rats. Thus, GPI 15427 and GPI 16539 exhibited anti inflammation activity against damage caused by intestinal ischemia/reperfusion and colitis. GPI 15427 and GPI 16539 may be useful for treating gut ischemia and inflammation. PMID- 16310768 TI - Behavioral and neural mechanisms of compulsive drug seeking. AB - Not the mere procurement and use of drugs, but the fact that patterns of seeking and taking become compulsive after prolonged drug use is a defining characteristic of drug addiction. Development of a therapy that targets the compulsive aspects of drug use and thus addresses addiction at its core would therefore be very desirable. In the present review, we will discuss animal studies that attempt to model loss of control over drug use. Furthermore, we will try to put these studies in a theoretical perspective, and discuss the hypothesized underlying neural and behavioral mechanisms. PMID- 16310769 TI - 5-HT1B receptors and aggression: a review. AB - The serotonergic (5-HT) system in the brain is involved in the modulation of offensive aggressive behavior. The dogma that activity of the 5-HT system is inversely related to aggression is obsolete now. Research on the status of the 5 HT system before, during and after the execution of aggression is ongoing but has not yet led to a clear picture about the actual functional role of the 5-HT system, the more because state versus trait aggression seems to play a pivotal role in the outcome. Pharmacological challenges pinpoint 5-HT(1A) and 5-HT(1B) receptors as key players in the modulation of offensive aggression. This review emphasizes in particular the role of postsynaptic 5-HT(1B) (hetero) receptors as a premier site to modulate offensive aggression. Modulation of the firing and 5 HT release of the serotonergic neuron, via presynaptic 5-HT(1A) (auto) receptors, presynaptic 5-HT(1B) (auto) receptors and serotonergic transporters, may also have striking influences on aggression under certain conditions. Therefore, it is hypothesized that postsynaptic 5-HT(1B) (hetero) receptors directly influence the executive, consummatory phases of agonistic behavior, whereas presynaptic serotonergic feedback systems are particularly useful in the introductory (appetitive) phases of the agonistic behavioral complex. PMID- 16310770 TI - Nucleocytoplasmic shuttling of STK16 (PKL12), a Golgi-resident serine/threonine kinase involved in VEGF expression regulation. AB - PKL12/STK16 protein is the first identified mammalian member of a ser/thr kinase subfamily that is conserved across several kingdoms, with a broad expression pattern in murine tissues and cell types. Endogenous STK16 subcellular localization was evaluated by indirect immunofluorescence in NIH/3T3 and NRK cells, demonstrating a Golgi-associated pattern that appears to be independent of signals provided by integrin pathways. When cells were treated with brefeldin A (BFA) or nocodazole, drugs that promote Golgi disorganization, we observed STK16 translocation to the nuclear compartment. Constitutive overexpression of this protein by retroviral vectors also promotes accumulation of STK16 in the nuclear compartment, as shown by subfractionation studies. A kinase-dead STK16 mutant (E202A) was used to demonstrate that both the Golgi association and the nuclear translocation capabilities seem to be independent of the STK16 kinase activity. In addition, we show that STK16 overexpression in several cell lines enhances their capacity to produce and secrete VEGF. To confirm these data in vivo, we injected tumor cells overexpressing STK16 into immunodeficient BALBc/SCID mice. HT1080-derived tumors overexpressing STK16 showed increased volume and number of blood vessels compared to controls. Altogether, these data concur with previous reports suggesting a potential role for STK16 as a transcriptional co-activator. PMID- 16310771 TI - VEGF-A, cytoskeletal dynamics, and the pathological vascular phenotype. AB - Normal angiogenesis is a complex process involving the organization of proliferating and migrating endothelial cells (ECs) into a well-ordered and highly functional vascular network. In contrast, pathological angiogenesis, which is a conspicuous feature of tumor growth, ischemic diseases, and chronic inflammation, is characterized by vessels with aberrant angioarchitecture and compromised barrier function. Herein we review the subject of pathological angiogenesis, particularly that driven by vascular endothelial growth factor (VEGF-A), from a new perspective. We propose that the serious structural and functional anomalies associated with VEGF-A-elicited neovessels, reflect, at least in part, imbalances in the internal molecular cues that govern the ordered assembly of ECs into three dimensional vascular networks and preserve vessel barrier function. Adopting such a viewpoint widens the focus from solely on specific pro-angiogenic stimuli such as VEGF-A to include a key set of cytoskeletal regulatory molecules, the Rho GTPases, which are known to direct multiple aspects of vascular morphogenesis including EC motility, alignment, multi-cellular organization, as well as intercellular junction integrity. We offer this perspective to draw attention to the importance of endothelial cytoskeletal dynamics for proper neovascularization and to suggest new therapeutic strategies with the potential to improve the pathological vascular phenotype. PMID- 16310772 TI - Alpha-synuclein is upregulated in neurones in response to chronic oxidative stress and is associated with neuroprotection. AB - Chronic oxidative stress has been linked to the neurodegenerative changes characteristic of Parkinson's disease, particularly alpha-synuclein accumulation and aggregation. However, it remains contentious whether these alpha-synuclein changes are cytotoxic or neuroprotective. The current study utilised long-term primary neural culture techniques with antioxidant free media to study the cellular response to chronic oxidative stress. Cells maintained in antioxidant free media were exquisitely more vulnerable to acute exposure to hydrogen peroxide, yet exposure of up to 10 days in antioxidant free media did not lead to morphological alterations in neurones or glia. However, a subpopulation of neurones demonstrated a significant increase in the level of alpha-synuclein expressed within the cell body and at synaptic sites. This subset of neurones was also more resistant to apoptotic changes following exposure to antioxidant free media relative to other neurones. These data indicate that increased alpha synuclein content is associated with neuroprotection from relatively low levels of oxidative stress. PMID- 16310773 TI - Neural progenitor implantation restores metabolic deficits in the brain following striatal quinolinic acid lesion. AB - Neural progenitor transplantation is a potential treatment for neurodegenerative diseases, including Huntington's disease (HD). In the current study, we tested the potential of rat embryonic neural progenitors expanded in vitro as therapy in the rat quinolinic acid-lesioned striatum, a model that demonstrates some of the pathological features of HD. We used positron emission tomography (PET) to demonstrate that the intrastriatal injection of cultured rat neural progenitors results in improved metabolic function in the striatum and overlying cortex when compared to media-injected controls. Transplanted progenitors were capable of surviving, migrating long distances and differentiating into neurons and glia. The cortices of transplanted animals contained greater numbers of neurons in regions that had shown metabolic improvement. However, histological analysis revealed that only a small fraction of these increased neurons could be accounted for by engrafted cells, indicating that the metabolic sparing was likely the result of a trophic action of the transplanted cells on the host. Behavioral testing of the implanted animals did not reveal improvement in apomorphine induced rotation. These data demonstrate that progenitor cell implantation results in enhanced metabolic function and sparing of neuron number, but that these functions do not necessarily result in the restoration of complex circuitry. PMID- 16310774 TI - Clonorchis sinensis and Opisthorchis viverrini: development of a mitochondrial based multiplex PCR for their identification and discrimination. AB - We report a single, one-step PCR approach for detection and discrimination of Clonorchis sinensis and Opisthorchis viverrini in different life-stage forms (adults, metacercariae, and eggs) from fish intermediate hosts and from infected patients. Primers designed for species-specific PCR, amplifying portions of the mitochondrial (mt) genome, were also suitable for a multiplex PCR. The latter was a single, one-step reaction under high stringency conditions, using simultaneously 2 pairs of primers (1 pair for C. sinensis--product size 612 bp, and 1 pair for O. viverrini--product size 1357 bp). Assays using serially diluted templates demonstrated that as little as 0.78 ng of genomic DNA of either species could yield amplicons. Genomic DNA extracted from different life-stage forms including adult worms (of both species), eggs (of O. viverrini), eggs possibly of several trematode species (collected from patients infected with C. sinensis in Vietnam) and mixed metacercariae of common trematodes (collected from fishes in the C. sinensis endemic areas), yielded specific bands of the correct size and their identity was confirmed by sequence analysis. The multiplex PCR approach described here proved to be a species-specific, sensitive and fast tool for accurate diagnosis of clonorchiasis and/or opisthorchiasis, permitting the detection of their metacercariae in infected fishes or adult/eggs from patients in endemic areas. PMID- 16310775 TI - Multiple incorporation of non-natural amino acids into a single protein using tRNAs with non-standard structures. AB - The ability to introduce non-natural amino acids into proteins opens up new vistas for the study of protein structure and function. This approach requires suppressor tRNAs that deliver the non-natural amino acid to a ribosome associated with an mRNA containing an expanded codon. The suppressor tRNAs must be absolutely protected from aminoacylation by any of the aminoacyl-tRNA synthetases in the protein synthesizing system, or a natural amino acid will be incorporated instead of the non-natural amino acid. Here, we found that some tRNAs with non standard structures could work as efficient four-base suppressors fulfilling the above orthogonal conditions. Using these tRNAs, we successfully demonstrated incorporation of three different non-natural amino acids into a single protein. PMID- 16310776 TI - Identification of multiple allosteric sites on the M1 muscarinic acetylcholine receptor. AB - Staurosporine and four staurosporine derivatives were docked on the rhodopsin based homology model of the M1 muscarinic acetylcholine receptor in order to localize the possible allosteric sites of this receptor. It was found that there were three major allosteric sites, two of which are located at the extracellular face of the receptor, and one in the intracellular domain of the receptor. In the present study, the localization of these binding sites is described for the first time. The present study confirms the existence of multiple allosteric sites on the M1 muscarinic receptor, and lays the ground for further experimental and computational analysis to better understand how muscarinic receptors are modulated via their allosteric sites. These findings will also help to design and develop novel drugs acting as allosteric modulators of the M1 receptor, which can be used in the treatment of the Alzheimer's disease. PMID- 16310777 TI - The binding of mycolic acids to galectin-3: a novel interaction between a host soluble lectin and trafficking mycobacterial lipids? AB - Understanding the molecular mechanism of host-pathogen interactions is the basis for drug design and vaccine development. The fine composition of mycolic acids (MA), the major constituents of Mycobacterium tuberculosis (Mtb) cell envelope, as well as other cell wall-associated lipids, contribute to determine the virulence of a given strain. However, endogenous receptors for mycolic acids on susceptible cells exposed to mycobacterial infections have not been fully identified. Here, we show that galectin-3, a multifunctional beta-galactoside binding lectin present mainly in the cytoplasm of inflammatory cells and also present on the cell surface, can recognize mycobacterial mycolic acids. MA can inhibit the lectin self-association but not its carbohydrate-binding abilities and can selectively interfere in the interaction of the lectin with its receptors on temperature-sensitive dendritic cell line, suggesting that galectin-3 could be involved in the recognition of trafficking mycolic acids and participate in their interaction with host cells. PMID- 16310778 TI - Adenovirus-mediated expression of the C-terminal domain of SARS-CoV spike protein is sufficient to induce apoptosis in Vero E6 cells. AB - The pro-apoptotic properties of severe acute respiratory syndrome coronavirus (SARS-CoV) structural proteins were studied in vitro. By monitoring apoptosis indicators including chromatin condensation, cellular DNA fragmentation and cell membrane asymmetry, we demonstrated that the adenovirus-mediated over-expression of SARS-CoV spike (S) protein and its C-terminal domain (S2) induce apoptosis in Vero E6 cells in a time- and dosage-dependent manner, whereas the expression of its N-terminal domain (S1) and other structural proteins, including envelope (E), membrane (M) and nucleocapsid (N) protein do not. These findings suggest a possible role of S and S2 protein in SARS-CoV induced apoptosis and the molecular pathogenesis of SARS. PMID- 16310780 TI - 5-Hydroxytryptamine as a potent migration enhancer of human aortic endothelial cells. AB - The purpose of the present study was to investigate whether 5-hydroxytryptamine (5-HT, serotonin) affects migration of vascular endothelial cells. 5-HT significantly enhanced migration of human aortic endothelial cells (HAECs), and this enhancement was completely inhibited by GR 55562, a 5-HT1 receptor antagonist, and fluoxetine, a 5-HT transporter inhibitor, but was not affected by ketanserin, a 5-HT2 receptor antagonist. 5-HT stimulation increased RhoA and ERK activity of HAECs, and inhibitors of RhoA (Y-27632 and H-1152) and inhibitors of MEK (U0126 and PD98059) abolished the 5-HT-induced increase in migration velocity. Inhibition of Rho kinase by Y-27632 blocked stress fiber formation and rear release of HAECs. Thus, 5-HT has a potent enhancing action on migration of HAECs through activating the RhoA and ERK pathways following 5-HT1 receptor stimulation. PMID- 16310781 TI - Structural basis for the specificity of basic winged bean lectin for the Tn antigen: a crystallographic, thermodynamic and modelling study. AB - The crystal structure of winged bean basic agglutinin in complex with GalNAc alpha-O-Ser (Tn-antigen) has been elucidated at 2.35 angstroms resolution in order to characterize the mode of binding of Tn-antigen with the lectin. The Gal moiety occupies the primary binding site and makes interactions similar to those found in other Gal/GalNAc specific legume lectins. The nitrogen and oxygen atoms of the acetamido group of the sugar make two hydrogen bonds with the protein atoms whereas its methyl group is stabilized by hydrophobic interactions. A water bridge formed between the terminal oxygen atoms of the serine residue of the Tn antigen and the side chain oxygen atom of Asn128 of the lectin increase the affinity of the lectin for Tn-antigen compared to that for GalNAc. A comparison with the available structures reveals that while the interactions of the glyconic part of the antigen are conserved, the mode of stabilization of the serine residue differs and depends on the nature of the protein residues in its vicinity. The structure provides a qualitative explanation for the thermodynamic parameters of the complexation of the lectin with Tn-antigen. Modeling studies indicate the possibility of an additional hydrogen bond with the lectin when the antigen is part of a glycoprotein. PMID- 16310782 TI - Transgenic potato expressing Abeta reduce Abeta burden in Alzheimer's disease mouse model. AB - Beta amyloid (Abeta) is believed one of the major pathogens of Alzheimer's disease (AD), and the reduction of Abeta is considered a primary therapeutic target. Immunization with Abeta can reduce Abeta burden and pathological features in transgenic AD model mice. Transgenic potato plants were made using genes encoding 5 tandem repeats of Abeta1-42 peptides with an ER retention signal. Amyloid precursor protein transgenic mice (Tg2576) fed with transgenic potato tubers with adjuvant showed a primary immune response and a partial reduction of Abeta burden in the brain. Thus, Abeta tandem repeats can be expressed in transgenic potato plants to form immunologically functional Abeta, and these potatoes has a potential to be used for the prevention and treatment of AD. PMID- 16310783 TI - The putative protein 6 of the severe acute respiratory syndrome-associated coronavirus: expression and functional characterization. AB - The SARS-CoV open reading frame 6 (ORF6) is transcribed into mRNA6 and encodes a putative 7.5 kDa accessory protein, SARS 6, with unknown function. In this study, we have confirmed the SARS 6 protein expression in lung and intestine tissues of the SARS patients and in SARS-CoV infected Vero E6 cells by immunohistochemistry. Further studies by immunoblot and confocal microscopy analyses revealed the expression and the endoplasmic reticulum (ER) localization of the recombinant SARS 6 protein in mammalian cells. Expression of SARS 6 protein in mammalian cells elicits biological activity of stimulating cellular DNA synthesis. PMID- 16310784 TI - Degradation of human aquaporin 0 by m-calpain. AB - Opacities (cataracts) in the lens of the eye are a leading cause of preventable blindness. Aquaporins function as water channels, and the C-terminus is postulated as a regulatory domain. The C-terminal domain of aquaporin 0 (AQP0) develops numerous truncation sites during lens aging. The purpose of the present experiment was to determine if the calcium-activated protease m-calpain (EC 3.4.22.17) was responsible for truncation of human AQP0. AQP0 was isolated from young human donors, incubated with recombinant m-calpain, and the cleavage sites on the released peptides were determined by on-line electrospray ionization mass spectrometry. We found that four cleavage sites on human AQP0 could be tentatively assigned to m-calpain. This is the first evidence for possible calpain activity in human lens. Because the cause(s) of 17 other cleavage sites was unknown, the data also suggested that other, as yet unknown, proteases or non enzymatic mechanisms are more active than calpain in human lens. PMID- 16310785 TI - Functional sulfurtransferase is associated with mitochondrial complex I from Yarrowia lipolytica, but is not required for assembly of its iron-sulfur clusters. AB - Here, we report that in the obligate aerobic yeast Yarrowia lipolytica, a protein exhibiting rhodanese (thiosulfate:cyanide sulfurtransferase) activity is associated with proton pumping NADH:ubiquinone oxidoreductase (complex I). Complex I is a key enzyme of the mitochondrial respiratory chain that contains eight iron-sulfur clusters. From a rhodanese deletion strain, we purified functional complex I that lacked the additional protein but was fully assembled and displayed no functional defects or changes in EPR signature. In contrast to previous suggestions, this indicated that the sulfurtransferase associated with Y. lipolytica complex I is not required for assembly of its iron-sulfur clusters. PMID- 16310786 TI - The cysteine-desulfurase IscS promotes the production of the rhodanese RhdA in the persulfurated form. AB - After heterologous expression in Escherichia coli, the Azotobacter vinelandii rhodanese RhdA is purified in a persulfurated form (RhdA-SSH). We identified l cysteine as the most effective sulfur source in producing RhdA-SSH. An E. coli soluble extract was required for in vitro persulfuration of RhdA, and the addition of pyridoxal-5'-phosphate increased RhdA-SSH production, indicating a likely involvement of a cysteine desulfurase. We were able to show the formation of a covalent complex between IscS and RhdA. By combining a time-course fluorescence assay and mass spectrometry analysis, we demonstrated the transfer of sulfur from E. coli IscS to RhdA. PMID- 16310787 TI - Localization of the nuclear receptor CAR at the cell membrane of mouse liver. AB - The nuclear receptor constitutive active/androstane receptor CAR is a drug sensing transcription factor. Upon activation by various drugs such as phenobarbital (PB), CAR translocates from the cytoplasm into the nucleus to regulate the genes that encode enzymes and proteins involved in hepatic metabolism. Here, we have shown the presence of CAR at the cell membrane of mouse livers, using Car+/+ and Car-/- mice. Levels of the cell membrane CAR increased after PB treatment. The CAR exists as a large approximately 160 kDa complex. Thus, CAR undergoes PB-induced translocation to the cell membrane, indicating that CAR may exert a non-genomic action. PMID- 16310788 TI - Evolution of host specificity in fleas: is it directional and irreversible? AB - Evolutionary trends in the evolution of host specificity have been the focus of much discussion but little rigorous empirical testing. On the one hand, specialization is often presumed to lead irreversibly into evolutionary dead ends and little diversification; this would mean that generalists might evolve into specialists, but not vice versa. On the other hand, low host specificity may limit the risk of extinction and provide more immediate fitness benefits to parasites, such that selection may favour evolution toward a generalist strategy. Here, we test for directionality in the evolution of host specificity using a large data set and phylogenetic information on 297 species of fleas parasitic on small mammals. The analyses determined whether host specificity, measured both as the number of host species exploited and their taxonomic diversity, was related to clade rank of the flea species, or the number of branching events between an extant species and the root of the phylogenetic tree (i.e., the total path length from the root of the tree to the species). Based on regression analyses, we found positive relationships between the number of host species used and clade rank across all 297 species, as well as within one (Hystrichopsyllidae) of four large families and one of seven large genera investigated separately; in addition, we found a positive relationship between the taxonomic diversity of host species used and clade rank in another of the seven genera. These results suggest a slight evolutionary trend of decreasing host specificity. Using a much more conservative likelihood ratio test, however, a random walk, or null model, of evolution could not be discarded in favour of the directional trends in all cases mentioned above. Still, these results suggest that host specificity may have tended to decrease in many flea lineages, a process that could have been driven by the benefits of exploiting a wide range of host species. PMID- 16310789 TI - Phytomonas serpens: cysteine peptidase inhibitors interfere with growth, ultrastructure and host adhesion. AB - In this study, we report the ultrastructural and growth alterations caused by cysteine peptidase inhibitors on the plant trypanosomatid Phytomonas serpens. We showed that the cysteine peptidase inhibitors at 10 microM were able to arrest cellular growth as well as promote alterations in the cell morphology, including the parasites becoming short and round. Additionally, iodoacetamide induced ultrastructural alterations, such as disintegration of cytoplasmic organelles, swelling of the nucleus and kinetoplast-mitochondrion complex, which culminated in parasite death. Leupeptin and antipain induced the appearance of microvillar extensions and blebs on the cytoplasmic membrane, resembling a shedding process. A 40 kDa cysteine peptidase was detected in hydrophobic and hydrophilic phases of P. serpens cells after Triton X-114 extraction. Additionally, we have shown through immunoblotting that anti-cruzipain polyclonal antibodies recognised two major polypeptides in P. serpens, including a 40 kDa component. Flow cytometry analysis confirmed that this cruzipain-like protein has a location on the cell surface. Ultrastructural immunocytochemical analysis demonstrated the presence of the cruzipain-like protein on the surface and in small membrane fragments released from leupeptin-treated parasites. Furthermore, the involvement of cysteine peptidases of P. serpens in the interaction with explanted salivary glands of the phytophagous insect Oncopeltus fasciatus was also investigated. When P. serpens cells were pre-treated with either cysteine peptidase inhibitors or anti-cruzipain antibody, a significant reduction of the interaction process was observed. Collectively, these results suggest that cysteine peptidases participate in several biological processes in P. serpens including cell growth and interaction with the invertebrate vector. PMID- 16310790 TI - Compatibility in the Biomphalaria glabrata/Echinostoma caproni model: potential involvement of adhesion genes. AB - Because susceptibility or resistance of Biomphalaria glabrata to the trematode Echinostoma caproni correlates with differential hemocytic adhesive properties, we compared the expression of genes involved in adhesion processes between hemocytes from susceptible and resistant snails. Quantitative reverse transcriptase-PCR analysis revealed four genes whose transcripts were differentially represented between hemocytes from resistant and susceptible snails. These genes encode two dermatopontin-like, one matrilin-like and one cadherin-like proteins. Expression analyses performed following parasite exposure suggested that dermatopontins may be involved in the compatibility differences between these strains. We also investigated expression levels on whole snails of different genes potentially involved in extracellular matrix structure or coagulation. Our results support the hypothesis that susceptible snails possess a hemolymph coagulation-like system that is more potent than that of resistant snails. This system may prevent hemocyte migration towards the parasite larvae and therefore facilitate parasite settlement in susceptible snails. PMID- 16310791 TI - Increased soluble FcgammaRIIIa(Mphi) in plasma from patients with coronary artery diseases. AB - Atherosclerosis is the underlying disease process in patients affected with coronary artery diseases (CAD). Macrophages play a major role in the development of vascular lesions in atherogenesis. The cells express Fcgamma receptor type IIIa (FcgammaRIIIa: CD16) identical to that in natural killer cells (NK cells), but with a cell type-specific glycosylation. In contrast, neutrophils express FcgammaRIIIb. These FcgammaRIIIs are released from the cell surface on activation, and these soluble forms (sFcgammaRIII) are present in the plasma. We measured sFcgammaRIIIa(Mphi) in the plasma with a newly developed anti FcgammaRIII mAb, MKGR14, which recognizes FcgammaRIIIa(Mphi) specifically. The level of sFcgammaRIIIa(Mphi), as well as the level of sFcgammaRIIIa (sFcgammaRIIIa(Mphi) plus sFcgammaRIIIa(NK)) or the level of total sFcgammaRIII (sFcgammaRIIIa plus sFcgammaRIIIb), were significantly increased in patients with CAD, but not in patients with vasospastic angina (VSA) or intact coronary arteries, compared with age-matched healthy donors. The sFcgammaRIIIa(Mphi) level was related to the number of significantly affected coronary arteries, and positively correlated with LDL-cholesterol to HDL-cholesterol ratios, but negatively with HDL-cholesterol. No correlation among the levels of three sFcgammaRIIIs was observed in CAD patients, as well as in healthy donors. The macrophages are activated during the process of atherosclerosis, and sFcgammaRIIIa(Mphi) may serve as a novel marker for atherosclerosis. PMID- 16310792 TI - An investigation by electron microscopy of chylomicron remnant uptake by human monocyte-derived macrophages. AB - Human monocyte-derived macrophages (HMM) internalise proatherogenic chylomicron remnants via several high affinity receptor pathways. However, the endocytic ultrastructures responsible for the uptake of chylomicron remnants by macrophages have not previously been described. In this study, we have utilised transmission electron microscopy together with colloidal gold-labelling of chylomicron remnants to investigate the pathways involved in macrophage uptake of chylomicron remnants. We found that macrophages internalise chylomicron remnants via surface connected compartments of up to 600 nm as well as non-clathrin coated pits. Chylomicron remnants were found to be distributed internally in a number of endocytic vesicles including early cysternal endosomes, spherical late endosomes and tubular vesicular compartments. Uptake of chylomicron remnants by HMM via phagocytosis or macropinocytosis was excluded based on the observations that lipoproteins were not found in phagolysosomes nor modified by inhibitors of these two processes, respectively. The latter observation contrasts with previous reports of chylomicron remnant internalisation by macrophages of other species. PMID- 16310793 TI - Control of ground reaction forces by hindlimb muscles during cat locomotion. AB - It has been proposed that biarticular muscles are primarily responsible for the control of the direction of external forces, as their activation is closely related and highly sensitive to the direction of external forces. This functional role for biarticular muscles has been supported qualitatively by experimental evidence, but has never been tested quantitatively for lack of a mathematical/mechanical formulation of this theory and the difficulty of measuring individual muscle forces during voluntary movements. The purposes of this study were: (1) to define rules for muscular coordination based on the control of external forces; (2) to develop a model of the cat hindlimb that allows for the calculation of the magnitude and direction of the ground reaction forces (GRFs) produced by individual hindlimb muscles; and (3) to test if the coordination of mono- and biarticular cat hindlimb muscles is related to the control of the resultant GRF. We measured the GRF, hindlimb kinematics, selected muscle forces and activations during cat locomotion. Then, the measured muscle forces were used as input to the hindlimb model to compute the muscle-induced GRF. We assume that if activation (and possibly force) increased as the muscle induced component of GRF approximated the resultant GRF, then that muscle was used by the central nervous system (CNS) to help control the direction of the external GRF. During cat walking, medial gastrocnemius (MG) and plantaris (PL) forces increased with increasing proximity to the GRF, while soleus (SOL) forces and vastus lateralis (VL) activations did not. SOL and VL activation were most strongly related to the vertical and parallel (braking/accelerating) component of the GRF, respectively. We concluded from these results that MG and PL are primarily responsible for the control of the direction of the GRF, while SOL primarily functions as an anti-gravity muscle, and VL as an acceleration/deceleration muscle. PMID- 16310794 TI - Rapid inorganic ion analysis using quantitative microchip capillary electrophoresis. AB - Rapid quantitative microchip capillary electrophoresis (CE) for online monitoring of drinking water enabling inorganic ion separation in less than 15 s is presented. Comparing cationic and anionic standards at different concentrations the analysis of cationic species resulted in non-linear calibration curves. We interpret this effect as a variation in the volume of the injected sample plug caused by changes of the electroosmotic flow (EOF) due to the strong interaction of bivalent cations with the glass surface. This explanation is supported by the observation of severe peak tailing. Conducting microchip CE analysis in a glass microchannel, optimized conditions are received for the cationic species K+, Na+, Ca2+, Mg2+ using a background electrolyte consisting of 30 mmol/L histidine and 2 (N-morpholino)ethanesulfonic acid, containing 0.5 mmol/L potassium chloride to reduce surface interaction and 4 mmol/L tartaric acid as a complexing agent resulting in a pH-value of 5.8. Applying reversed EOF co-migration for the anionic species Cl-, SO42- and HCO3- optimized separation occurs in a background electrolyte consisting of 10 mmol/L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 10 mmol/L HEPES sodium salt, containing 0.05 mmol/L CTAB (cetyltrimethylammonium bromide) resulting in a pH-value of 7.5. The detection limits are 20 micromol/L for the monovalent cationic and anionic species and 10 micromol/L for the divalent species. These values make the method very suitable for many applications including the analysis of abundant ions in tap water as demonstrated in this paper. PMID- 16310795 TI - Screening of nitrogen mustards and their degradation products in water and decontamination solution by liquid chromatography-mass spectrometry. AB - Analysing nitrogen mustards and their degradation products in decontamination emulsions posed a significant challenge due to the different phases present in such matrices. Extensive sample preparation may be required to isolate target analytes. Furthermore, numerous reaction products are formed in the decontamination emulsion. A fast and effective qualitative screening procedure was developed for these compounds, using liquid chromatography-mass spectrometry (LC-MS). This eliminated the need for additional sample handling and derivatisation that are required for gas chromatographic-mass spectrometric (GC MS) analysis. A liquid chromatograph with mixed mode column and isocratic elution gave good chromatography. The feasibility of applying this technique for detecting these compounds in spiked water and decontamination emulsion was demonstrated. Detailed characterisation of the degradation products in these two matrices was carried out. The results demonstrated that N-methyldiethanolamine (MDEA), N-ethyldiethanolamine (EDEA) and triethanolamine (TEA) are not the major degradation products of their respective nitrogen mustards. Degradation profiles of nitrogen mustards in water were also established. In verification analysis, it is important not only to develop methods for the identification of the actual chemical agents; the methods must also encompass degradation products of the chemical agents as well so as to exclude false negatives. This study demonstrated the increasingly pivotal role that LC-MS play in verification analysis. PMID- 16310796 TI - Capillary liquid chromatography-tandem mass spectrometry of tetrahydroisoquinoline derived neurotoxins: a study on the blood-brain barrier of rat brain. AB - Certain tetrahydroisoquinoline derivatives such as 1-benzyl-1,2,3,4 tetrahydroisoquinoline (1-BnTIQ) and N-methylsalsolinol are parkinsonian neurotoxins. This paper describes a sensitive and reliable analytical method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the determination of tetrahydroisoquinoline derivatives (TIQs) in brain dialysate. Samples (20 microL injected) were effectively stacked and cleaned up in-line on a capillary column (5 cm x 0.25 mm I.D.) packed with 5 microm phenyl reversed-phase silica particles. Under the optimized conditions, electrospray ionisation-MS/MS detection of TIQs was highly sensitive. The capillary LC-MS/MS method had a detection limit of 2 ng/ml for TIQ. The method was used in combination with in vivo microdialysis to study the blood-brain barrier (BBB) for TIQs. The microdialysis probe was implanted in the frontal cortex of rat brain. Test compounds were administered intraperitoneally (i.p.). Four TIQs including 1,2,3,4 tetrahydroisoquinoline (TIQ), 5,6,7,8-tetrahydroisoquinoline (5-TIQ), 1-BnTIQ, and salsolinol (SAL) were studied. A concentration maximum was detected in brain dialysate for TIQ, 5-TIQ, and 1-BnTIQ about 40 min after drug administration. However, SAL, the precursor of N-methylsalsolinol was found unable to cross the BBB of rat brain. PMID- 16310797 TI - Identification of alkyl carbazoles and alkyl benzocarbazoles in Brazilian petroleum derivatives. AB - Carbozoles are important compounds in crude oils, as they may be used as geochemical tracers, being the major type of nitrogen compounds in petroleum. At the same time, they are regarded as undesirable due to the problems they may cause in the refining process, such as catalyst poisoning, corrosion, gum or color formation in final products. As separation and identification of carbazoles are challenging goals, this work presents a chromatographic method, made of a pre fractionation on neutral alumina followed by the separation and identification of two classes of carbazoles using FeCl(3)/Chromossorb W and gas chromatograph with mass spectrometer (GC/MS) (SIM-single ion monitoring mode) analysis. For the first time, a series of alkyl carbazoles and alkyl benzocarbazoles were identified in heavy gas oil (HGO) and atmospheric residue of distillation (ARD) obtained from Brazilian petroleum. PMID- 16310798 TI - Diffusion coefficient of Brownian particle in rough micro-channel. AB - This study examines the feasibility of using of the lattice Boltzmann method to determine how the surface roughness of a quadrate channel affects the diffusion coefficient of Brownian particle(s). The surface was represented by a regular array of spheres. Surface roughness reduced the diffusion coefficient of the Brownian particle(s) because of a change in the velocity autocorrelation function decay and in pressure. Additionally, the neighboring particles increased the diffusion coefficient of Brownian particle. PMID- 16310799 TI - Comparison of the surface characteristics of polypropylene films treated by Ar and mixed gas (Ar/O2) atmospheric pressure plasma. AB - In an attempt to modify the hydrophobic surface properties of polypropylene (PP) films, this study examined the optimum process parameters of atmospheric pressure plasma (APP) using Ar gas. Under optimized conditions, the effects of a mixed gas (Ar/O2) plasma treatment on the surface-free energy of a PP film were investigated as a function of the O2 content. The polar contribution of the surface-free energy of the PP film increased with increasing O2 content in the gas mixture. However, slightly more oxygen-containing polar functional groups such as CO, CO, and COO were introduced on the PP film surface by the Ar gas only rather than by the Ar/O2 gas mixture. In addition, AFM analysis showed that the Ar plasma treatment of the PP film produced the smoothest surface as a result of the relatively homogeneous etching process. PMID- 16310800 TI - Cooperativity in the adsorption of magnetic colloidal particles. AB - In this paper we investigate the adsorption of magnetic particles onto magnetically patterned substrates. We find that the adsorption process is cooperative, where the probability of adsorption decreases with increasing substrate occupancy (namely, density of adsorbed particles). The effect of cooperativity can be accounted for by a simple modification of the adsorption probability as manifested by the binomial distribution. The negative cooperativity found in the magnetic particle adsorption is not due to direct repulsion between particles, but to screening of the surface's magnetic field by previously adsorbed particles. Thus, the adsorption of magnetic colloids on magnetic substrates is a self-limiting process. PMID- 16310801 TI - A novel sandwich ELISA for alpha1 domain based detection of soluble HLA-G heavy chains. AB - The detection of soluble human leukocyte antigen G (HLA-G) has been a technically demanding task for several years now and various enzyme linked immunosorbent assay (ELISA) formats have been designed. However, no ELISA test has been described so far which is able to detect all possible kinds of soluble HLA-G (sHLA-G) molecules that might occur in bio fluids. Here we describe a new ELISA approach able to recognize soluble alpha1 domain containing heavy chains of all HLA-G isoforms. The detection limit is shown to be at about 150 pg soluble recombinant HLA-G1 heavy chain per milliliters. Detectable HLA-G fragments are shown to occur in the supernatants of different HLA-G transfected cell lines and appear to be particularly abundant in supernatant of trophoblast derived choriocarcinoma cell lines. The novel ELISA employs the well characterized HLA-G mAbs 4H84 and MEM-G1 which ensure high HLA-G specificity. A negative control ELISA format, designed against non-existing analytes, has been established to reveal non-specific signal interference. PMID- 16310802 TI - Engineering of large numbers of highly specific homing endonucleases that induce recombination on novel DNA targets. AB - The last decade has seen the emergence of a universal method for precise and efficient genome engineering. This method relies on the use of sequence-specific endonucleases such as homing endonucleases. The structures of several of these proteins are known, allowing for site-directed mutagenesis of residues essential for DNA binding. Here, we show that a semi-rational approach can be used to derive hundreds of novel proteins from I-CreI, a homing endonuclease from the LAGLIDADG family. These novel endonucleases display a wide range of cleavage patterns in yeast and mammalian cells that in most cases are highly specific and distinct from I-CreI. Second, rules for protein/DNA interaction can be inferred from statistical analysis. Third, novel endonucleases can be combined to create heterodimeric protein species, thereby greatly enhancing the number of potential targets. These results describe a straightforward approach for engineering novel endonucleases with tailored specificities, while preserving the activity and specificity of natural homing endonucleases, and thereby deliver new tools for genome engineering. PMID- 16310803 TI - Three-dimensional structure of a double apoptosome formed by the Drosophila Apaf 1 related killer. AB - The Drosophila Apaf-1 related killer (Dark) forms an apoptosome that activates Dronc, an apical procaspase in the intrinsic cell death pathway. To study this process, we assembled a large Dark complex in the presence of dATP. Remarkably, we found that cytochrome c was not required for assembly and when added, cytochrome c did not bind to the Dark complex. We then determined a 3D structure of the Dark complex at 18.8A resolution using electron cryo-microscopy and single particle methods. In the structure, eight Dark subunits form a wheel-like particle and two of these rings associate face-to-face. In contrast, Apaf-1 forms a single ring that is comprised of seven subunits and each Apaf-1 binds a molecule of cytochrome c. We then used relevant crystal structures to model the Dark complex. This analysis shows that a single Dark ring and the Apaf-1 apoptosome share many key features. When taken together, the data suggest that a single ring in the Dark complex may represent the Drosophila apoptosome. Thus, our analysis provides a domain model of this complex and gives insights into its function. PMID- 16310804 TI - Safety-culture in a Norwegian shipping company. AB - PROBLEM: Although there has been considerable interest in safety culture and safety climate in many industries, little attention has been given to safety culture in one of the world's riskiest industries, shipping. METHOD: Using both self developed items and items from published research on safety culture, safety climate, and quality and management style, a 40-item safety culture questionnaire was developed. The questionnaire was distributed in a self-administered form to sailors onboard 20 vessels and to officers attending a seminar in Manila. A total of 349 questionnaires were collected (total response rate, 60%). RESULTS, DISCUSSION AND IMPACT ON INDUSTRY: Principal component analysis (PCA) revealed 11 factors when the Kaiser eigenvalue rule was used and four factors when the scree test criterion was used. The factor structure in the material confirmed structures found in other industries. The relative importance of the factors from the factor analysis on "level of safety" measures was tested by canonical correlation analysis and regression analysis. The results confirmed previous research and showed that the most important factors were influential across industries. To determine weather differences existed between nationalities, occupations, and vessels the factors from the PCA was subjected to Multiple Discriminant Analysis. Significant differences between occupations, nations, and vessels were found on one or more of the factors from the PCA. PMID- 16310805 TI - Meiotic CAG repeat instability in spinocerebellar ataxia type 6: maternally transmitted elongation in a presumed sporadic case. AB - Spinocerebellar ataxia type 6 (SCA6) is an autosomal dominantly inherited disorder characterized by cerebellar ataxia, dysarthria and nystagmus. The molecular background for the disorder is a CAG repeat expansion in the CACNA1A gene located on chromosome 19. The size of SCA6 expanded alleles is usually stable, and variation in repeat size over successive generations is rare. We report a Danish family with one case of SCA6 resembling a sporadic case of spinocerebellar ataxia. Analysis of the CACNA1A gene showed meiotic CAG repeat instability in the transmission from a 70-year-old woman with no subjective symptoms to her symptomatic son. The CAG repeat size expanded from 22 repeats in the mother to 23 repeats in the proband. This case demonstrates maternal repeat instability and clinical anticipation in a family with SCA6. PMID- 16310806 TI - Proterguride, a highly potent dopamine receptor agonist promising for transdermal administration in Parkinson's disease: interactions with alpha(1)-, 5-HT(2)- and H(1)-receptors. AB - Dopamine receptor agonists play an important role in the treatment of Parkinson's disease and hyperprolactinemic conditions. Proterguride (n-propyldihydrolisuride) was already reported to be a highly potent dopamine receptor agonist, thus its action at different non-dopaminergic monoamine receptors, alpha(1A/1B/1D), 5 HT(2A/2B)- and histamine H(1), was investigated using different functional in vitro assays. The drug behaved as an antagonist at alpha(1)-adrenoceptors without the ability to discriminate between the subtypes (pA(2) values: alpha(1A) 7.31; alpha(1B) 7.37; alpha(1D) 7.35) and showed antagonistic properties at the histamine H(1) receptor. In contrast, at serotonergic receptors (5-HT(2A), 5 HT(2B)) proterguride acted as a partial agonist. The drug stimulated 5-HT(2A) receptors of rat tail artery in lower concentrations than 5-HT itself but failed to evoke comparable efficacy (proterguride: pEC(50) 8.34, E(max) 53% related to the maximum response to 5-HT; 5-HT: pEC(50) 7.03). Agonism at 5-HT(2B) receptors is presently considered to be involved in drug-induced valvular heart disease. Activation of 5-HT(2B) receptors in porcine pulmonary arteries by proterguride (pEC(50) 7.13, E(max) 49%; E(max) (5-HT) 69%), however, occurred at concentrations much higher than plasma concentrations achieving dopaminergic efficacy in humans. The results are discussed focussing on the relevance of action at 5-HT(2B) receptors as well as their significance for a transdermal administration of proterguride. Since it is well accepted that pulsatile dopaminergic stimulation is associated with treatment-related motor complications in the dopaminergic therapy of Parkinson's disease, the transdermal route of administration is of great clinical interest due to the possibility to achieve constant plasma concentrations. PMID- 16310807 TI - Induction of G2/M phase arrest by squamocin in chronic myeloid leukemia (K562) cells. AB - Squamocin is one of the annonaceous acetogenins and has been reported to have anticancer activity. Squamocin was found to inhibit the growth of K562 cells in a time- and dose-dependent manner. Cell cycle analysis showed G2/M phase arrest in K562 cells following 24 h exposure to squamocin. During the G2/M arrest, cyclin dependent kinase inhibitors (CDKIs), p21 and p27 were increased in a dose dependent manner. Analysis of the cell cycle regulatory proteins demonstrated that squamocin did not change the steady-state levels of Cdk2, Cdk4, cyclin A, cyclin B1, cyclin D3 and cyclin E, but decreased the protein levels of Cdk1 and Cdc25C. These results suggest that squamocin inhibits the proliferation of K562 cells via G2/M arrest in association with the induction of p21, p27 and the reduction of Cdk1 and Cdc25C kinase activities. PMID- 16310808 TI - Effect of thalidomide affecting VEGF secretion, cell migration, adhesion and capillary tube formation of human endothelial EA.hy 926 cells. AB - Angiogenesis, new blood vessel formation, is a multistep process, precisely regulated by pro-angiogenic cytokines, which stimulate endothelial cells to migrate, proliferate and differentiate to form new capillary microvessels. Excessive vascular development and blood vessel remodeling appears in psoriasis, rheumatoid arthritis, diabetic retinopathy and solid tumors formation. Thalidomide [alpha-(N-phthalimido)-glutarimide] is known to be a potent inhibitor of angiogenesis, but the mechanism of its inhibitory action remains unclear. The aim of the study was to investigate the potential influence of thalidomide on the several steps of angiogenesis, using in vitro models. We have evaluated the effect of thalidomide on VEGF secretion, cell migration, adhesion as well as in capillary formation of human endothelial cell line EA.hy 926. Thalidomide at the concentrations of 0.01 microM and 10 microM inhibited VEGF secretion into supernatants, decreased the number of formed capillary tubes and increased cell adhesion to collagen. Administration of thalidomide at the concentration of 0.01 microM increased cell migration, while at 10 microM, it decreased cell migration. Thalidomide in concentrations from 0.1 microM to 10 microM did not change cell proliferation of 72-h cell cultures. We conclude that anti-angiogenic action of thalidomide is due to direct inhibitory action on VEGF secretion and capillary microvessel formation as well as immunomodulatory influence on EA.hy 926 cells migration and adhesion. PMID- 16310809 TI - General administration of cyclohexenonic long-chain fatty alcohol ameliorates hyperreactivity of STZ-induced diabetic rat aorta. AB - Diabetic neuropathy, a major complication of diabetes mellitus, is associated with the development of vascular dysfunction and autonomic neuropathy. We studied the effects of cyclohexenonic long-chain fatty alcohol (FA) on streptozotocin diabetic hyperreactivity in the rat aorta smooth muscle. The rats were divided randomly into four groups and were maintained for 4 weeks: age-matched control rats, diabetic rats without treatment with FA, and diabetic rats treated with FA (2 and 8 mg/kg, i.p. everyday). The serum glucose and insulin levels were determined, and the contractile responses of the aorta induced by a thromboxane A2 agonist, U46619 and KCl were investigated. Treatment with FA did not alter rats' diabetic status, i.e., body weight, thickness of the aorta, serum glucose levels, and serum insulin levels, but significantly improved the diabetic-induced hyperreactivity of the rat aorta in a dose-dependent manner. Removal of endothelium did not change contractile force between groups. In histological examinations, thinning of smooth muscle bundle in the wall of aorta was observed in the diabetic rat, which was not significantly improved by treatment with FA. Our data indicate that FA can prevent hyperreactivity in the diabetic aorta. PMID- 16310810 TI - Molecular modeling of SWR-0342SA, a beta3-selective agonist, with beta1- and beta3-adrenoceptor. AB - The molecular dynamics (MD) simulations study in the formation of the complex between compound SWR-0342SA and beta-ARs suggested that upon binding SWR-0342SA stimulates receptor activation through residues network (Asp104, Leu335 in beta(1)-AR; Asp117, Ser209, Leu303, Ser191 in beta(3)-AR) in an active conformation state. The models suggest that the structural origin of the selectivity of SWR-0342SA to beta(3)-AR vs. beta(1)-AR comes from the following results: (a) the tight interaction between the agonist and the TMs 3, 5, 6 and 2 nd EC loop. Asp117 interacts with the cationic amino group of the agonist molecule. (b) Additional contacts are done with Ser209, Leu303 and Ser191. These results are in good agreement with the binding affinities (pKi values) of SWR 0342SA to beta-AR family expressed in recombinant mammalian cells. PMID- 16310811 TI - A single-molecule PCR approach to the measurement of induced expanded simple tandem repeat instability in vitro. AB - Sensitive and precise models are needed to identify potential genotoxicity at environmentally relevant doses of mutagens. The size length alterations in expanded simple tandem repeat (ESTR) loci have been used as a biomarker of genetic instability caused by a variety of agents in the mouse germline. The mechanisms operating in both spontaneous and induced instability are poorly understood. We have developed a single-molecule polymerase chain reaction (SM PCR) method to investigate mutation at the mouse ESTR locus Ms6-hm in the murine C3H/10T1/2 embryonic cell line. Growth of cells to levels of high cell density induced increased ESTR instability, with mutation frequencies 5.1-fold (+/-2.8) over sub-confluent cultures. Accordingly, cell cultures were maintained at sub confluent levels for further investigations of the induction of ESTR mutation by genotoxic agents. Treatment with the DNA alkylating agent N-nitroso-N-ethylurea (ENU) resulted in a 1.94-fold (+/-1.1) increase in mutation frequency, similar to responses measured previously in the germline in vivo. Therefore, mutagen exposure can also affect somatic (non-meiotic) rapidly dividing mouse cells. This SM-PCR approach eliminates the requirement of sub-cloning individual treated cells, thereby, reducing the time needed to screen for ESTR mutation, and will be a very useful tool for future investigations into the mechanisms involved in ESTR mutation. PMID- 16310812 TI - Effects of histamine H(3) antagonists and donepezil on learning and mnemonic deficits induced by pentylenetetrazol kindling in weanling mice. AB - Childhood epilepsy is one of the main risk factors for a variety of problems involving cognition and behavior. Pentylenetetrazol (PTZ) kindling is currently an acceptable model for epilepsy research. The objectives of this study are to clarify the learning and mnemonic characteristics of PTZ kindling in developing mice, and to examine the effects of thioperamide and JNJ-5207852, two histamine H(3) receptor antagonists and donepezil, an acetylcholinesterase (AChE) inhibitor, on learning and memory deficits induced by PTZ kindling in the brains of developing mice. PTZ kindling led to learning and mnemonic deficits as assessed by social discrimination, acoustic fear conditioning, water maze and passive avoidance tests. Thioperamide and JNJ-5207852, ameliorated PTZ kindling induced learning and mnemonic deficits in all tests except for the water maze test. In addition, the learning and mnemonic impairments induced by PTZ kindling were significantly improved by donepezil in all tests. These findings suggest that histamine and acetylcholine are involved in the different processes of learning and memory in the brain and that histamine H(3) receptor antagonists might be useful in the treatment of cognitive impairment in epilepsy. PMID- 16310813 TI - Neural correlates of cognitive control in childhood and adolescence: disentangling the contributions of age and executive function. AB - Dense-array (128-channel) electroencephalography (EEG) was used to record event related potentials (ERPs) from 33 participants between 7 and 16 years of age while they performed a Go/Nogo task. The frontal (Nogo) N2 component of the ERP was taken as an index of cognitive control, and examined in relation to both age and independent assessments of executive function (EF), including the Iowa Gambling Task (IGT), the Stroop task, a delay discounting task, and backward digit span. Better performance on the IGT and the Stroop task was associated with smaller N2 amplitudes, over and above effects of age. N2 latencies decreased with age but were not predicted by EF. Source modeling of the N2 revealed neural generators in areas suggestive of cingulate cortex and orbitofrontal cortex, and the locations of these generators varied systematically with EF (IGT and Stroop task): the cingulate generator was more anterior for good EF participants at all ages; the orbitofrontal generator was relatively left lateralized for younger and for poorer EF participants. Taken together, these findings suggest that age related decreases in N2 amplitude, but not N2 latency, reflect the development of cognitive control and cannot be attributed solely to incidental changes that may affect assessments of the N2 (e.g., increases in skull thickness). Functionally relevant decreases in N2 amplitude may reflect changes in the regions of cortex giving rise to the N2. PMID- 16310814 TI - Heredity influences the dietary energy density of free-living humans. AB - Dietary energy density has been shown to have large effects on short-term food intake. Little is known, however, about its origins. We investigated the influence of heredity, shared (familial) environment, and individual environment on dietary energy density and its relationship to food intake with 110 identical and 102 fraternal same-sex and 53 fraternal mixed gender adult twin pairs who were paid to maintain 7-day food intake diaries. From the diary reports, the total and meal intakes of food energy, the amounts of the macronutrients ingested, and dietary energy density were estimated. Linear structural modeling was applied to investigate the nature and degree of genetic and environmental influences. The analysis revealed significant genetic and individual environmental, but not shared (familial) environmental, influences on the dietary densities of the reported diets. But, there was no genetic or familial influences on the responsiveness of the individual to dietary density. Dietary energy density appears to be another component in a package of genetically determined physiological, sociocultural, and psychological processes that influence the regulation of energy balance. PMID- 16310815 TI - Scrapie resistance alleles are not associated with lower prolificity in Rasa Aragonesa sheep. AB - Scrapie is a prion disease characterised by the accumulation of the pathological associated form of cellular prion protein (PrP(SC)) in the central nervous system. Susceptibility to scrapie is associated with polymorphism in the ovine prion protein (PrP) gene. The European Union has implemented scrapie control programs, relying on selective breeding for scrapie resistance; the use of ARR carrier and the exclusion of VRQ-carrier were recommended. In this study, 4323 individuals from Rasa Aragonesa Sheep breed were genotyped for the PrP gene and the individual estimated breeding values (EBV) for prolificity were calculated. Most represented PrP alleles do not work against prolificity. Only a significant association between VRQ/VRQ genotype and a lower EBV was observed (p = 0.027, eta2 = 0.002). Therefore, avoiding reproduction of VRQ/VRQ individuals would not cause negative effect regarding prolificity. PMID- 16310816 TI - Changes in peripheral leukocytes enzymes activity and plasma metabolite concentrations in growing Holstein calves. AB - Plasma metabolite and immunoreactive insulin concentrations and activities of enzymes related to energy metabolism in peripheral leukocytes were measured in growing Holstein calves. A ratio of girth of abdomen divided by girth of thorax (A/T ratio) of calves was significantly elevated after weaning, and the A/T ratio maybe a good indicator to evaluate rumen development. Plasma glucose and free fatty acid concentrations were changed in calves accompanying change in feeding. Activities of lactate dehydrogenase with pyruvate as substrate (LDH-P) and hexokinase (HK) in cytosolic fractions of peripheral leukocytes decreased significantly after weaning the calves reflecting the change of energy source from milk replacer with high percentages of fat and glucose and lactose as absorbable carbohydrate to pelleted feed containing starch as less absorbable carbohydrate and roughage. Some peripheral leukocyte enzymes such as LDH and HK may be good indicators to evaluate changes in energy metabolism of growing calves. PMID- 16310817 TI - Monoacylglycerol lipase inhibition by organophosphorus compounds leads to elevation of brain 2-arachidonoylglycerol and the associated hypomotility in mice. AB - Three components of the cannabinoid system are sensitive to selected organophosphorus (OP) compounds: monoacylglycerol (MAG) lipase that hydrolyzes the major endogenous agonist 2-arachidonoylglycerol (2-AG); fatty acid amide hydrolase (FAAH) that cleaves the agonist anandamide present in smaller amounts; the CB1 receptor itself. This investigation considers which component of the cannabinoid system is the most likely contributor to OP-induced hypomotility in mice. Structure-activity studies by our laboratory and others rule against major involvement of a direct toxicant-CB1 receptor interaction for selected OPs. Attention was therefore focused on the OP sensitivities of MAG lipase and FAAH, assaying 19 structurally diverse OP chemicals (pesticides, their metabolites and designer compounds) for in vitro inhibition of both enzymes. Remarkably high potency and low selectivity is observed with three O-alkyl (C1, C2, C3) alkylphosphonofluoridates (C8, C12) (IC50 0.60-3.0 nM), five S-alkyl (C5, C7, C9) and alkyl (C10, C12) benzodioxaphosphorin oxides (IC50 0.15-5.7 nM) and one OP insecticide metabolite (chlorpyrifos oxon, IC50 34-40 nM). In ip-treated mice, the OPs at 1-30 mg/kg more potently inhibit brain FAAH than MAG lipase, but FAAH inhibition is not correlated with hypomotility. However, the alkylphosphonofluoridate-treated mice show dose-dependent increases in severity of hypomotility, inhibition of MAG lipase activity and elevation of 2-AG. Moderate to severe hypomotility is accompanied by 64 to 86% MAG lipase inhibition and about 6-fold elevation of brain 2-AG level. It therefore appears that OP induced MAG lipase inhibition leads to elevated 2-AG and the associated hypomotility. PMID- 16310818 TI - Functional structure of Agelena labyrinthica's (Araneae:Agelenidae) venom gland and electrophoresis of venom. AB - The funnel-web spider, Agelena labyrinthica, is widely distributed throughout Turkey. The objective of the present study was to describe the histological and functional fine structure of A. labyrinthica's venom gland by using light microscope, scanning (SEM) and transmission electron microscope (TEM). We have also preliminarily analyzed venom components by SDS-PAGE. Each venom gland has surrounded by a thin adventitia and gross striated muscular bundles. Basal lamina underlies between muscular bundles and the inner glandular epithelium, and ties up them each other. The striated muscular bundles spirally covered venom gland has been observed by SEM. Intricate relations formed between motor neuron axons and the muscle fibers have been revealed by TEM. The secretory epithelium, which made up of simple columnar cells, formed the secretory region of the venom gland. The secretory surface of the gland was increased by a sort of fringes extended from basal membrane into the gland lumen. The epithelial cells have many rough endoplasmic reticulum, mitochondria and different size and shape of secretory granules. These granules have been accumulated in apical portion of the secretory cells. After the gland is emptied, the apical portions of secretory cells deteriorate and the basal epithelial cells regenerate the columnar cells. The analysis of A. labyrinthica venom, which was achieved by SDS-PAGE showed that there have been at least seven components ranging from 10 to 40 kDa molecular weight. PMID- 16310819 TI - Acute renal failure provoked by toxin from caterpillars of the species Lonomia obliqua. AB - Acute renal failure (ARF) is a complication of envenoming by contact with caterpillars of the species Lonomia obliqua. A cohort study was conducted to evaluate development of acute and chronic renal failure (CRF) in patients who been in contact with L. obliqua during the period from 1989 to 2003 in the State of Santa Catarina, southern Brazil. Patients were evaluated in two distinct groups: those prior to 1995, who did not receive specific treatment of any type, and those after this date who were treated with antilonomic serum (SALon). The presence of creatinine > or = 1.5 mg/dl in patients who had no history of previous renal illness was considered to be indicative of acute kidney failure. Of the 2067 patients evaluated, 39 (1.9%) developed ARF. Eleven (32%) of these patients were treated with dialysis and four (10.3%) developed CRF. The seven deaths (4%) occurred in the period before treatment with SALon. Blood coagulation measures (TC, TP, and TTPA), were significantly longer in the group with ARF. All patients with ARF and 67% of the control group presented hematuria. The majority of the patients recovered prior renal function, however, four (10.3%) needed chronic dialysis. Envenoming by L. obliqua can be considered an important risk factor for the development of potentially fatal ARF, as well as for developing CRF. PMID- 16310820 TI - MMTV accessory factor Naf affects cellular gene expression. AB - Mouse mammary tumour virus (MMTV) encodes a viral superantigen (Sag) and a negative acting factor (Naf) which share parts of their coding sequence. Using 2 dimensional gel electrophoresis (2D-DIGE), we could show that at least 10 different cellular proteins were differentially expressed in Naf positive cells. Also, luciferase reporter expression was down-regulated in Naf expressing cells independent of the promoter used and further experiments suggested that this effect was due in part to a decrease in cellular growth rates. Although in Naf positive cells expression of the major sag containing transcript was strongly induced by the synthetic glucocorticoid dexamethasone, the hormone analogue neither influenced luciferase expression nor mRNA expression of selected cellular proteins identified by 2D-DIGE. Taken together, these data support the previous finding that Naf and Sag have separable activities and suggest that Naf may play a role in modulating host cell gene expression during MMTV infection. PMID- 16310821 TI - Adsorbent obtained from CEPT sludge in wastewater chemically enhanced treatment. AB - Chemically enhanced primary treatment (CEPT) in municipal wastewater treatment is particularly suitable for rapidly growing cities. The focus on CEPT process now might be the cost of chemicals and disposal of chemical sludge. In this study, the potential application of adsorbent made from CEPT sludge in CEPT wastewater treatment was investigated in various conditions, such as the adsorbent dosage, pH, and dosing modes of ferric chloride and adsorbent. It was found that sludge derived adsorbent was a good way of sludge disposal while enhanced pollutants removal rate and reduced the fresh ferric chloride dosage. With the use of 10.0 mg L(-1) of ferric chloride and 0.6 g L(-1) of the adsorbent at the first stage simultaneously, the enhanced removal efficiencies of turbidity, UV254, COD, TP were 83.3%, 52.3%, 48.8% and 89.0%, respectively. The experiments showed that ferric chloride dose was reduced about 50%. The pH played a significant role in coagulation and adsorption process. PMID- 16310822 TI - Correlations between arsenic in Maine groundwater and microbial populations as determined by fluorescence in situ hybridization. AB - Arsenic is known to cause serious health effects when consumed in drinking water. In the state of Maine, approximately half of the population relies on private groundwater wells for their drinking water. Of those wells, as many as 13% may contain arsenic levels above the current EPA maximum contaminant level of 10 microgl(-1). Microorganisms can potentially contribute to arsenic release into groundwater through several mechanisms. Some can reduce arsenate to arsenite, which is more toxic and may be more mobile. Sulfurospirillum species NP4, which was isolated from well water, respires arsenate and could act in this way. Microorganisms can also act indirectly by reducing bedrock surface coatings, such as iron oxyhydroxides, that adsorb arsenic in the groundwater environment. The genus Geobacter contains many species that are capable of iron reduction that could play a role in the indirect release of arsenic into groundwater. Water samples from Northport, ME and the Branch Lake region of Ellsworth, ME, which both have elevated groundwater arsenic levels, have been probed using fluorescence in situ hybridization (FISH), to determine the percentage of the population that is NP4 and the percentage that are Geobacter species. Geobacter abundance correlates well with the total arsenic concentration indicating that indirect mechanisms could be important in releasing arsenic. NP4 appears to be reducing arsenate since its prevalence correlates well with arsenite, the end product of arsenate respiration. PMID- 16310823 TI - Fungal and enzymatic decolourisation of artificial textile dye baths. AB - A textile dye Reactive Black 5 was used in screening 25 fungal strains for their decolourising ability. The most promising strains were tested in a medium containing specific constituents of a dye bath in order to approach real application conditions. It was shown that the concentrations of the constituents had to be reduced to allow fungal growth. Decolourisation started in cultures of Geotrichum candidum but was not complete. Only Bjerkandera adusta was able to decolourise the black-blue colour through violet and red to pale yellow. After 17 days spectral absorption coefficients, alpha, at three wavelengths, 620, 525 and 436 nm almost reached the permitted values. A partly purified manganese peroxidase prepared from B. adusta was tested for decolourisation of several artificial dye baths. The constituents seemed not to be inhibitory to the enzyme and no dilution was necessary. Evaluation of decolourisation gave different results, depending on the method used. The most efficient decolourisation on a percentage basis was observed in the dye bath of the anthraquinone dye Reactive Blue 19, followed by the diazo dye Reactive Black 5. However, based on absorbance units, the largest reduction was achieved with the Reactive Black 5 and Acid Orange 7 dye baths. Comparing the alpha values after 120 h fungal and enzymatic treatments of Reactive Black 5 dye bath the enzyme showed about 1.5 times greater colour reduction than the fungus. Given the tolerance to the constituents and concentration of dye baths, the enzyme proved to be a promising tool for their treatment. PMID- 16310824 TI - Mechanically induced self-propagating reactions: analysis of reactive substrates and degradation of aromatic sulfonic pollutants. AB - Mechanochemical reactions have been identified as a valuable alternative to conventional methodologies for the degradation of toxic pollutants as well as for their abatement in contaminated matrices. This paper discusses the application of the mechanochemical technique to the degradation of sulfonic acids in a contaminated matrix. The degradation of the pollutant compound was carried out by taking advantage of combustive reactions on a suitable reactive system ignited under mechanical treatment conditions. Two systems have been investigated as possible reactive substrates. The first one was a Mg-SiO2 powder mixture while the second system was a Ca-SiO2 powder mixture. Milling trials performed under different mechanical processing conditions allowed one to characterise the reactivity of these chemical systems, which basically undergo a reduction/oxidation reaction involving the formation of MgO and Si phases when the Mg-SiO2 system is considered and CaO and Si phases when the Ca-SiO2 system is employed, respectively. The systematic change of the stoichiometric ratios Mg:SiO2 and Ca:SiO2 permitted to identify the minimum Mg or Ca content necessary for the ignition of the combustive reactions. The experimental runs performed with such systems have shown a greater effectiveness of the Mg-SiO2 because of less energy inputs required to ignite a combustion. For this reason the Mg-SiO2 has been considered as a reactive substrate in the following trials. Since the SiO2 amount in stoichiometric excess may be regarded as inert phase, it was substituted with a different phase consisting of the matrix contaminated by sulfonic acids. This aspect permitted to ignite a combustive reaction with the minimum possible content of Mg-SiO2 reacting mixture. The chemical analyses performed after the combustive reaction proved the complete removal of the sulfonic acid from the contaminated matrix. PMID- 16310825 TI - Adaptive neuro-fuzzy based modelling for prediction of air pollution daily levels in city of Zonguldak. AB - Air pollution is a growing problem arising from domestic heating, high density of vehicle traffic, electricity production, and expanding commercial and industrial activities, all increasing in parallel with urban population. Monitoring and forecasting of air quality parameters in the urban area are important due to health impact. Artificial intelligent techniques are successfully used in modelling of highly complex and non-linear phenomena. In this study, adaptive neuro-fuzzy logic method has been proposed to estimate the impact of meteorological factors on SO2 and total suspended particular matter (TSP) pollution levels over an urban area. The model forecasts satisfactorily the trends in SO2 and TSP concentration levels, with performance between 75-90% and 69-80 %, respectively. PMID- 16310826 TI - Microbial indicators of heavy metal contamination in urban and rural soils. AB - Urban soils and especially their microbiology have been a neglected area of study. In this paper, we report on microbial properties of urban soils compared to rural soils of similar lithogenic origin in the vicinity of Aberdeen city. Significant differences in basal respiration rates, microbial biomass and ecophysiological parameters were found in urban soils compared to rural soils. Analysis of community level physiological profiles (CLPP) of micro-organisms showed they consumed C sources faster in urban soils to maintain the same level activity as those in rural soils. Cu, Pb, Zn and Ni were the principal elements that had accumulated in urban soils compared with their rural counterparts with Pb being the most significant metal to distinguish urban soils from rural soils. Sequential extraction showed the final residue after extraction was normally the highest proportion except for Pb, for which the hydroxylamine-hydrochloride extractable Pb was the largest part. Acetic acid extractable fraction of Cd, Cu, Ni, Pb and Zn were higher in urban soils and aqua regia extractable fraction were lower suggesting an elevated availability of heavy metals in urban soils. Correlation analyses between different microbial indicators (basal respiration, biomass-C, and sole C source tests) and heavy metal fractions indicated that basal respiration was negatively correlated with soil Cd, Cu, Ni and Zn inputs while soil microbial biomass was only significantly correlated with Pb. However, both exchangeable and iron- and manganese-bound Ni fractions were mostly responsible for shift of the soil microbial community level physiological profiles (sole C source tests). These data suggest soil microbial indicators can be useful indicators of pollutant heavy metal stress on the health of urban soils. PMID- 16310827 TI - Molluscicidal activity of Ferula asafoetida, Syzygium aromaticum and Carum carvi and their active components against the snail Lymnaea acuminata. AB - The molluscicidal activity of dried root latex powder of Ferula asafoetida, flower-bud powder of Syzygium aromaticum and seed powder of Carum carvi against the snail Lymnaea acuminata was studied. The molluscicidal activity of all the three plant products was found to be both time and concentration dependent. The toxicity of S. aromaticum flower-bud powder (96 h LC(50):51.98 mg/l) was more pronounced than that of root latex powder of F. asafoetida (96 h LC(50):82.71 mg/l) and seed powder of C. carvi (96 h LC(50):140.58 mg/l). Ethanol extract was more toxic than other organic extracts. The ethanol extract of S. aromaticum (24h LC(50):83.53 mg/l) was more effective than that of F. asafoetida (24h LC(50):132.31 mg/l) and C. carvi (24h LC(50):130.61 mg/l) in killing the test animals. The 96 h LC(50) of column purified fraction of seed powder of C. carvi was 5.40 mg/l whereas those of flower-bud powder of S. aromaticum and dried root latex powder of F. asafoetida were 7.87 and 9.67 mg/l, respectively. The product of F. asafoetida, S. aromaticum and C. carvi may be used as potent molluscicides. PMID- 16310828 TI - Effect of soil chemical properties on the remediation of phenanthrene contaminated soil by electrokinetic-Fenton process. AB - The electrokinetic-Fenton (EK-Fenton) remediation of soil contaminated with phenanthrene was studied. Two different soils were chosen to investigate the effects of chemical properties, such as Fe oxide contents and acid soil buffer capacity. The H(2)O(2) concentrations in pore water, the electrical potential distributions and the electrical currents were monitored to assess the electrochemical effect in relation to the soil properties. Hadong caly had high acid buffer capacity, and thus the amount of electroosmotic flow was lager in the experiment with Hadong clay than with EPK kaolin. The major mechanism of phenanthrene removal was a degradation in the experiment with EPK Kaolin, while it was a simple transport away from the system in experiment with Hadong clay. It was mainly because of the lower acid buffering capacity and better H(2)O(2) stability in case with EPK Kaolin than with Hadong clay. PMID- 16310829 TI - Cadmium concentration in tobacco (Nicotiana tabacum L.) from different countries and its relationship with other elements. AB - Cadmium accumulation in crop plants, such as tobacco (Nicotiana tabacum L.), can lead to human exposure to this carcinogenic metal. To better define actual Cd distribution in cured or processed tobacco leaves from chief tobacco-producing regions, we analyzed 755 leaf samples of three major types (Flue-cured, Burley, and Oriental) obtained from 13 countries during 2001-2003. This survey may help identify regions with low- or high-Cd concentrations in tobacco to obtain insight into the cause of the concentration extremes and to assist in defining strategies to reduce Cd in tobacco. Cadmium concentrations in the samples ranged from 0 to 6.78 microg g(-1), as determined by ICP-MS. Significant differences were found among types and among countries, but significant interactions between type and country were found. Variations in Cd concentrations were also found in all countries. Our results suggest an important contribution of the field (e.g., bioavailable Cd in soil, other soil characteristics) to the Cd concentration in tobacco. Finally, the correlation between the concentration of Cd and that of other elements differed among the types, which could be effectively discriminated based on the concentrations of 20 elements. PMID- 16310830 TI - The role of humic substances in chromium sorption onto natural organic matter (peat). AB - To elucidate mechanisms of Cr3+ sorption onto the unaltered solid natural organic matter, the comparative studies of this ion binding from a solution at pH 4.0 onto three selected particle size fractions: 2000-1000 microm, 630-200 microm and 63-20 microm of markedly different HS content and structure, separated by a wet sieving from an overall sample of peat (Brushwood Peat Humus) were carried out. Comparable patterns of COOH groups and CEC(t) confirmed that for cation exchange capacity were responsible mainly cations connected with COO- functional groups. It was though found that aliphatic acids in the solid state did not take part in Cr3+ binding, thus the finest studied fraction 63-20 microm of the highest contents of functional groups showed the lowest sorption capacity for Cr3+, while similar patterns of sorbed Cr3+, soluble HS content and base CEC(0) indicated that these parameters were directly interrelated. The base ion exchange processes determined by CEC(0) (with Ca2+ as a predominant exchangeable cation) appeared to be not the major mechanisms responsible for Cr3+ sorption. For this metal, strong binding to insoluble large molecular weight organic pool two- to threefold prevailed over the ion exchange processes. Very low acid desorption indicated generally low mobility of Cr3+ -organic compounds. PMID- 16310831 TI - Biphenyl degradation kinetics by Burkholderia xenovorans LB400 in two-phase partitioning bioreactors. AB - Biphenyl could be successfully degraded by Burkholderia xenovorans LB400, initially described as Pseudomonas sp. LB400, in two-phase partitioning bioreactors (TPPBs). TPPBs are comprised of an aqueous, cell containing phase, and an immiscible, biocompatible organic phase that partitions toxic and/or poorly soluble substrates (in this case biphenyl) based on maintaining a thermodynamic equilibrium. The critical LogKO/W of the organism was found to be approximately 5.5, indicating that solvents with a LogKO/W larger than 5.5 are suitable as delivery phases for B. xenovorans LB400. Two solvents selected for the TPPB system were octadecene and bis(2-ethylhexyl)sebacate (BES). In one experiment a total of 6.6 g biphenyl per l aqueous-phase-equivalent (biphenyl delivered in solvent, at an aqueous phase to solvent ratio of 10) could be degraded in 25 h during batch operation with octadecene. The specific growth rate and the half saturation constant of the Monod model were estimated to be mumax=0.25 h-1 and KS=0.0001 g l-1, and the yield coefficient was YX/S=0.48 g biomass per g biphenyl. These parameter estimates were used to predict the time course of biphenyl degradation at different initial substrate concentrations and with biphenyl delivered from the two solvents with different partitioning behaviour for biphenyl. The predictions were validated by experimental data, confirming the microbial kinetics as well as the expected partitioning effects. PMID- 16310832 TI - Equilibrium sorption of phenanthrene by soil humic acids. AB - This study investigated the effect of chemical heterogeneity of humic acids (HAs) on the equilibrium sorption of phenanthrene by HA extracts. Six HA samples were extracted from three different soils with 0.5 M NaOH and 0.1 M Na4P2O7 and were characterized with elemental analysis, infrared spectrometry, and solid-state 13C nuclear magnetic resonance (NMR) spectrometry. The equilibrium sorption measurements were carried out with a batch technique and using the six HA solids as the sorbents and phenanthrene as the sorbate. The measured sorption isotherm data were fitted to the Freundlich equation. The results showed that, for the same soil, (i) the total HA mass extracted with Na4P2O7 was 13.7-22.6% less than that extracted with NaOH, (ii) the Na4P2O7-extracted HA had higher O/C atomic ratio, greater content of polar organic carbons (POC), and lower aliphatic carbon content than the NaOH-extracted HA, and (iii) the Na4P2O7-extracted HA exhibited greater sorption isotherm linearity and but not dramatic difference in sorption capacities than the NaOH extracted HA. The differences in the HA properties resulting from the two different extraction methods may be because NaOH can hydrolyze insoluble HA fractions such as fatty acid like macromolecules bound on soils whereas Na4P2O7 could not. As a result, the HAs extracted with the two different methods had different polarity and functionality which affected their sorption property for phenanthrene. PMID- 16310833 TI - Limb bone bilateral asymmetry: variability and commonality among modern humans. AB - Humans demonstrate species-wide bilateral asymmetry in long bone dimensions. Previous studies have documented greater right-biases in upper limb bone dimensions--especially in length and diaphyseal breadth--as well as more asymmetry in the upper limb when compared with the lower limb. Some studies have reported left-bias in lower limb bone dimensions, which, combined with the contralateral asymmetry in upper limbs, has been termed "crossed symmetry." The examination of sexual dimorphism and population variation in asymmetry has been limited. This study re-examines these topics in a large, geographically and temporally diverse sample of 780 Holocene adult humans. Fourteen bilateral measures were taken, including maximum lengths, articular and peri-articular breadths, and diaphyseal breadths of the femur, tibia, humerus, and radius. Dimensions were converted into percentage directional (%DA) and absolute (%AA) asymmetries. Results reveal that average diaphyseal breadths in both the upper and lower limbs have the greatest absolute and directional asymmetry among all populations, with lower asymmetry evident in maximum lengths or articular dimensions. Upper limb bones demonstrate a systematic right-bias in all dimensions, while lower limb elements have biases closer to zero %DA, but with slight left-bias in diaphyseal breadths and femoral length. Crossed symmetry exists within individuals between similar dimensions of the upper and lower limbs. Females have more asymmetric and right-biased upper limb maximum lengths, while males have greater humeral diaphyseal and head breadth %DAs. The lower limb demonstrates little sexual dimorphism in asymmetry. Industrial groups exhibit relatively less asymmetry than pre-industrial humans and less dimorphism in asymmetry. A mixture of influences from both genetic and behavioral factors is implicated as the source of these patterns. PMID- 16310834 TI - Clinical and neuropsychological features of corticobasal degeneration. AB - In this paper, we review the history, clinical, cognitive, imaging and neuropathological features of corticobasal degeneration (CBD). Additionally, we report new findings on the neuropsychological profile of 14 patients with CBD compared to 14 with Alzheimer's disease. Increasing awareness of this disorder has led to the realization that specific cognitive and imaging changes are common with CBD degeneration and that these changes help with clinical diagnosis. PMID- 16310835 TI - Extracellular enzyme activity and dynamics of bacterial community in mucilaginous aggregates of the northern Adriatic Sea. AB - Bacterial degradation of mucilaginous aggregates (creamy layers, stringers and macroflocs) collected during two summer events (2001-2002) was tested. The objective was to describe the temporal trend of the bacterial activity, abundance and composition in the aggregated and dissolved organic matter under different trophic conditions. In the native aggregates proteins and organic phosphorous were actively hydrolyzed as aminopeptidase and alkaline phosphatase activities represented up to 87% and 25% of total activity, respectively; polysaccharides were less hydrolyzed and the highest activities were observed for beta glucosidase (5% of the total). This hydrolysation pattern tends to a progressive accumulation of long persistent polysaccharides. During short term incubations nutrient addition (P, N and Glucose) differently stimulated bacterial growth in the seawater: P played the main role in stimulating bacterial production from 3 to 6 folds higher than in the control, whereas a secondary C-limitation was observed only for bacteria growing on seawater from macroflocs. This scarce dissolved organic carbon (DOC) bioavailability was confirmed by the lower DOC removal (13% macroflocs, 36% stringers). The total amount of carbon incorporated by bacteria living on aggregates was similar (0.58 mg C L(-1)) both in the control and under P enrichments showing a more balanced condition with respect to the seawater. Hence the well-known P limitation in the Northern Adriatic Sea affects only dissolved organic carbon uptake without influencing the uptake of aggregated organic matter. Organic matter limitation was observed only on stringers--total C incorporated raised to 0.96 mg C L(-1) after PNG addition. Macroflocs release of refractory compounds leads to DOC accumulation (73 microM DOC) contributing to inflate the pool of refractory DOC in the surrounding waters. Several evidences, including different monosaccharide composition of stringers and macroflocs (glucose 15% and 56% on stringers and macroflocs, respectively), bring to the conclusion that stringers are in an older stage in comparison with macroflocs. Community composition described by fluorescence in situ hybridization did not show significant differences between free-living and attached bacteria but it was modified by the different enrichment conditions: Cytophaga-Flavobacteria increased after inorganic nutrients enrichments while organics advantaged gamma-Proteobacteria. PMID- 16310836 TI - Atmospheric mercury in Norway: contributions from different sources. AB - The environmental loadings of national Norwegian mercury emissions compared to the loadings of atmospheric long range transported mercury have been estimated using national emission data and EMEP model data. The results indicate that atmospheric long-range transport to Norway is somewhat larger than the national Norwegian emissions of mercury. Atmospheric deposition of mercury has been studied using data from Norwegian monitoring programs on mercury in precipitation, mosses, natural surface soils, and lake sediments. Precipitation data show no significant time trend during 1990-2002, whereas moss samples show similar concentrations from 1985 to 1995, but a 30% decrease from 1995 to 2000. Concentrations of mercury in peat cores and reference sediments indicate that the current mercury levels measured in surface sediments, surface soils and mosses at background sites in Norway are substantially affected by long-range atmospheric transport. PMID- 16310837 TI - Saquinavir does not affect adipose tissue development or metabolic parameters in mice on high fat diet. PMID- 16310838 TI - Optimal reversal of vitamin K antagonists. PMID- 16310839 TI - Adnexal masses in pregnancy: how often are they malignant? AB - OBJECTIVE: The primary objective was to investigate the occurrence rates of benign and malignant ovarian tumors associated with pregnancy among women identified in three large California databases between 1991 and 1999. The secondary objective was to determine maternal and perinatal outcomes among these pregnancies. METHODS: This is a population-based study of 4,846,505 obstetrical patients using California hospital discharge records from 1991-1999. The California vital statistics birth/patient discharge database was linked to the California Cancer Registry (CCR). Cases of maternal ovarian cancers and low malignant potential (LMP) tumors were separated into three periods based on the timing of diagnosis and pregnancy: prenatal, at delivery, and postpartum. International Classification of Diseases, Revision 9 (ICD-9) codes were used to identify both diagnostic and procedural factors occurring during hospitalizations. The CCR database was used to identify cancer outcomes such as stage, histology, treatments, and vital status. RESULTS: 9375 women had a hospital diagnosis of an ovarian mass associated with pregnancy. CCR database identified 87 ovarian cancers and 115 LMP tumors in the same cohort. The occurrence rates were 0.93% (87/9375) ovarian cancers per total number of ovarian masses diagnosed during pregnancy, and 0.0179 ovarian cancers per 1000 deliveries. The summary stages of the ovarian cancers and LMP tumors were (respectively): localized 65.5% and 81.7%, regional 6.9% and 7.8%, remote 23.0% and 4.4%, and unknown 4.6% and 6.1%. 34 of the 87 ovarian cancers were germ cell tumors (GCT). Malignant ovarian tumors increased the likelihood of maternal outcomes such as cesarean delivery, hysterectomy, blood transfusions, and prolonged hospitalization compared to noncancer pregnant controls, but did not adversely affect neonatal outcomes. Cause-specific maternal mortality of patients with follow-up was 4.7% (9/191) at a mean of 2.43 years after diagnosis. CONCLUSIONS: Ovarian malignancies are rare during pregnancy. Most maternal malignant ovarian neoplasms are early stage and associated with favorable maternal and neonatal outcomes. The low maternal mortality rate is likely due to the predominance of GCTs among the ovarian cancers. PMID- 16310840 TI - Effect of timing of prostaglandin administration, controlled internal drug release removal and gonadotropin releasing hormone administration on pregnancy rate in fixed-time AI protocols in crossbred Angus cows. AB - Two experiments were conducted to investigate the effects of timing of prostaglandin F2(alpha) (PGF2(alpha)) administration, controlled internal drug release device (CIDR) removal and second gonodotropin releasing hormone (GnRH) administration on the pregnancy outcome in CIDR-based synchronization protocols. In Experiment 1, suckled Angus crossbred beef cows (n = 580) were given 100 microg of GnRH+a CIDR on Day 0. Cows in Group 1 (modified Ovsynch-P) received 25 mg of dinoprost (PGF2(alpha)) and CIDR device removal on Day 8 (AM), 100 microg of GnRH 36 h later on Day 9 (p.m.), and fixed-time AI (FTAI) 16 h later on Day 10 (47.5+/-1.1 h after PGF2(alpha)). Cows in Group 2 (Ovsynch-P) received 25mg of PGF2(alpha) and CIDR device removal on Day 7 (p.m.), 100 microg of GnRH 48 h later on Day 9 and FTAI 16 h later on Day 10 (66.6+/-1.2 h after PGF2(alpha)). Pregnancy rates were 56.5% (170/301) for Group 1 and 55.6% (155/279) for Group 2, respectively (P = 0.47). In Experiment 2, beef cows (n=734) were synchronized with 100 microg of GnRH+CIDR on Day 0, 25 mg of PGF2(alpha) and CIDR device removal on Day 7 and either 100 microg of GnRH 48 h later on Day 9 (Ovsynch-P) and FTAI 16 h later on Day 10 (64.9+/-3.3 h from PGF2(alpha)) or 100 microg of GnRH on Day 10 (CO-Synch-P) at the time of AI (63.2+/-4.2 h from PGF2(alpha)). Pregnancy rates were 48.8% (180/369) for Ovsynch-P and 44.7% (163/365) for CO synch-P groups, respectively (P = 0.11). In both experiments, there was a locationxtreatment interaction (P<0.05); pregnancy rates between locations were different (P < 0.05) in the Ovsynch-P group. In conclusion, in a CIDR-based Ovsynch synchronization protocol, delaying administration of prostaglandin and CIDR removal by 12 h, or timing of the second GnRH by 16 h, did not affect pregnancy rates to FTAI. Therefore, there may be an opportunity to make changes in synchronization protocols with out adversely affecting FTAI pregnancy rates. PMID- 16310841 TI - Comparison between glycerol and ethylene glycol for dog semen cryopreservation. AB - The aim of this study was to compare the effect of ethylene glycol versus glycerol for dog semen freezing, on post-thaw longevity, motility and motility parameters, and on plasma membrane functional integrity. Semen was diluted in two steps with an egg yolk TRIS extender containing a final concentration of either 5% glycerol or 5% ethylene glycol, and frozen in 0.5 ml straws, with 100 x 10(6) spermatozoa/ml, over nitrogen vapours. Semen motility was evaluated both under a light microscope and with a Computer Assisted Motility Analyser System, immediately after thawing and then hourly till 4h of incubation. Sperm membrane functional integrity was assessed with the hypoosmotic swelling test (60 mOsm fructose solution) applied at thawing and then hourly, for 4 h, on incubated samples. Motility (light microscope) and total and progressive motility (analyser) were significantly higher in ethylene glycol frozen samples at thawing (P < 0.01); from hour 1 onwards the effect of the cryoprotectant became not significant. Semen frozen with ethylene glycol showed higher path velocity and higher straight line velocity till 3 h after thawing; however, ethylene glycol semen samples also showed higher curvilinear velocity and higher lateral head displacement, which may indicate a capacitation-like condition affecting sperm membranes and possibly reducing post-thaw longevity. Functional integrity of plasma membrane was similar in glycerol and ethylene glycol samples till 3 h after thawing, then ethylene glycol samples showed a higher decline. The strong though short-lived positive effect of ethylene glycol is worth being evaluated further. PMID- 16310842 TI - Improvement of parameters of freezing medium and freezing protocol for bull sperm using two osmotic supports. AB - The aim of this study was to improve the freezing protocol of bull sperm, by investigating the influence on sperm viability after freeze/thawing of different freezing medium components, as well as the effect of cooling rates in the different stages of the cooling protocol, in single factor experiments. The experimental variables were: (1) salt-based versus a sugar-based medium (Tris versus sucrose); (2) glycerol concentration; (3) detergent (Equex) concentration; (4) presence of bicarbonate; (5) rate of cooling from 22 degrees C to holding temperature (CR1); (6) holding temperature (HT); (7) rate of cooling from holding temperature to -6 degrees C (CR2); (8) rate of cooling from -10 to -100 degrees C (CR3). All experiments were performed using five bulls per experiment (three ejaculates per bull). Sperm motility after freezing and thawing was assessed by CASA system, and sperm membrane integrity was assessed by flow cytometry. Sucrose based medium did not offer a clear significant benefit compared to Tris medium. The concentration of Equex that gave the best results in Tris-based media group and sucrose-based media group was in a range between 2-7 and 4-7 g/l, respectively. In both media groups, a glycerol concentration of 800 mM was the best in any post-thaw viability parameters. In the Tris media group, the presence of bicarbonate had a negative effect on sperm viability. CR1 and CR2 had no significant effect on any of the post-thaw sperm viability parameters, but a CR1=0.2 degrees C/min and CR2=4 degrees C/min appeared to give better results in both media. The holding temperature (HT) that gave the best results was found to be in the range of 5-9 degrees C. There was a significant disadvantage of using a low CR3 of 10 degrees C/min, while 150 degrees C/min appeared to be the best cooling rate for either medium. PMID- 16310843 TI - Antioxidant activity of different sulfate content derivatives of polysaccharide extracted from Ulva pertusa (Chlorophyta) in vitro. AB - Polysaccharide extracted from Ulva pertusa (Chlorophyta) is a group of sulfated heteropolysaccharide; for simplicity, the sulfated polysaccharide is referred to as ulvan in this paper. In this study, different sulfate content ulvans were prepared with sulfur trioxide/N,N-dimethylformamide (SO3-DMF) in formamide, and their antioxidant activities were investigated including scavenging activity of superoxide and hydroxyl radicals, reducing power and metal chelating ability. As expected, we obtained several satisfying results, as follows: firstly, high sulfate content ulvans had more effective scavenging activity on hydroxyl radical than natural ulvan. Secondly, comparing with natural ulvan, high sulfate content ulvans exhibited stronger reducing power. Thirdly, HU4 (sulfate content, 30.8%) and HU5 (sulfate content, 32.8%) showed more pronounce chelating ability on ferrous ion at high concentration than other samples. PMID- 16310844 TI - Nanopowder molding method for creating implantable high-aspect-ratio electrodes on thin flexible substrates. AB - Metal nanoparticles and a nanopowder molding process were used to fabricate 2D and 3D patternable structures having a height-to-width ratio of up to 10:1. By means of this process, an entire neural stimulation circuit, including stimulating electrode, connection trace, and contact pad, can be fused into one continuous, integrated structure where different sections can have different heights, widths, and shapes. The technique is suitable for mass production, and the fabricated electrode is robust and very flexible. More importantly for biomedical applications, the entire fabricated structure can be packed at room temperature onto a biocompatible flexible substrate, such as polydimethylsiloxane, parylene, and polyimide as well as other temperature sensitive or vacuum-sensitive materials. The electrodes and wires have about the same electrical resistivities as bulk materials and desirable electrochemical properties, including low impedance. PMID- 16310845 TI - Combinatorial investigation of the structure-properties characterization of photopolymerized dimethacrylate networks. AB - The effects of co-monomer composition and irradiation time in a model two component dimethacrylate dental resin blend were evaluated using combinatorial methods to determine the degree of methacrylate conversion and resulting mechanical properties. 2-Dimensional gradient samples varying in monomer composition and light exposure time were fabricated. The conversion was measured using near infrared spectroscopy (NIR) and the mechanical properties (i.e., hardness and elastic modulus) were determined using nanoindentation via the continuous stiffness method. An excellent correlation was observed between the reaction conversion and mechanical properties for the cross-linked networks. The methacrylate conversion ranged from 40% to 85% and the mechanical properties increased over two orders of magnitude over this conversion range. The ultimate reaction conversion and mechanical properties depended on both the co-monomer composition and cure time. PMID- 16310846 TI - The enzymatic degradation of scaffolds and their replacement by vascularized extracellular matrix in the murine myocardium. AB - Replacement of injured myocardium by cell-based degradable scaffolds is a novel approach to regenerate myocardium. Understanding the foreign body reaction (FBR) induced by the scaffold is requisite to predict unwanted site effects or implant failure. We evaluated the FBR against a biodegradable scaffold applied on injured myocardium in mice. Cryolesions and collagen type I scaffolds (Col-I) were applied to the left ventricle of mice. Cell infiltration, neovascularization, collagen deposition, matrix metalloproteinase (MMP-8) expression, enzymatic activity and scaffold degradation were determined at different time points (2-70 days). Infiltration of mainly macrophages, neutrophils and blood vessels was completed within 14 days. High numbers of neutrophils accumulated around the Col I fibers and degradation of Col-I fibers into small fragments was observed on day 14. Active MMP-8 co-localized with the neutrophils on day 14, indicating enzymatic degradation of Col-I by neutrophil collagenase. Highly vascularized extracellular matrix remained at day 70. No differences were observed in the FBR to Col-I after application on healthy or injured myocardium. The FBR had no adverse effects on the adjacent myocardial tissue. In conclusion, cardiac scaffolds are degraded by MMP-8 and replaced by vascularized extracellular matrix during the FBR on injured myocardium. PMID- 16310847 TI - Collagen-based implants reinforced by chitin fibres in a goat shank bone defect model. AB - Tissue engineering is an increasingly popular method for repairing bone defects. However, repair of bone defects over 30 mm using tissue-engineering methods is a difficult clinical problem. In this study, we used a goat shank model to evaluate the bone-regenerating efficacy of a novel nano-hydroxyapatite/collagen/PLLA (nHACP) composite reinforced by chitin fibres. Forty adult male goats with 40 mm defects in shank at the same anatomic site were divided into four groups. The first group was the control, where nothing was implanted in the defect (defect group). The other three groups were implanted with porous pure PLLA, nHACP and nHACP reinforced by chitin fibres, respectively. Bone growth in each group was evaluated by radiography, histology, bone mineral density and mechanical strength, once every 5 weeks for 15 weeks. The results indicated that nHACP implants, both with and without chitin fibres, are better for repairing the defects than pure PLLA. However, only the reinforced implants showed nearly perfect recovery in 15 weeks after operation. So, the reinforced scaffold might be a candidate for bone tissue repair. PMID- 16310848 TI - Mechanical properties of bacterial cellulose and interactions with smooth muscle cells. AB - Tissue engineered blood vessels (TEBV) represent an attractive approach for overcoming reconstructive problems associated with vascular diseases by providing small calibre vascular grafts. The aim of this study has been to evaluate a novel biomaterial, bacterial cellulose (BC), as a potential scaffold for TEBV. The morphology of the BC pellicle grown in static culture was investigated with SEM. Mechanical properties of BC were measured in Krebs solution and compared with the properties of porcine carotid arteries and ePTFE grafts. Attachment, proliferation and ingrowth of human smooth muscle cells (SMC) on the BC were analysed in vitro. The BC pellicle had an asymmetric structure composed of a fine network of nanofibrils similar to a collagen network. The shape of the stress strain response of BC is reminiscent of the stress-strain response of the carotid artery, most probably due to the similarity in architecture of the nanofibrill networks. SMC adhered to and proliferated on the BC pellicle; an ingrowth of up to 40 microm was seen after 2 weeks of culture. BC exhibit attractive properties for use in future TEBV. PMID- 16310849 TI - PEO-PPO-PEO-based poly(ether ester urethane)s as degradable reverse thermo responsive multiblock copolymers. AB - Aiming at developing biodegradable thermo-responsive polymers that display enhanced rheological properties, a family of PEO-PPO-PEO based poly(ether ester urethane)s, was developed. The materials were produced following a two-step synthetic pathway. The PEO-PPO-PEO triblocks were first end-capped with LA or CL oligo(ester)s whereby pentablocks were produced. Then, the different precursors were chain extended using hexamethylene diisocyanate to create the respective polymers. The length and type of the ester block influenced the behavior of the molecules in water, especially their viscosity versus temperature response. The gelation temperature increased from 23 degrees C for a 20wt% F127 solution to 26 and 31 degrees C for pentablocks with 4.4 and 7.5 lactoyl units, respectively. Materials containing longer LA units failed to show any reverse thermo responsiveness. The presence of the oligo(ester) blocks also reduced the viscosity of the gel at 37 degrees C. While F127 displayed a viscosity of around 28,000Pas, pentablocks containing 4.4 and 7.5 LA units showed values of 15,400 and 12,600Pas. Also, the viscosity at 37 degrees C as well as the gelation temperature decreased as the molecular weight of the oligo(ester)s increased. Finally, the degradation process of the gels was studied by monitoring their viscosity at body temperature and determining the molecular weight of the polymers, over time. Polymers were tailored so to combine high initial viscosity values with diverse degradation rates, as a function of the length and type of the oligo(ester) present along the polymeric backbone. PMID- 16310850 TI - Control of smooth muscle cell proliferation by ferrous iron. AB - This study was conducted to determine the interaction of individual corrosion products from biodegradable iron stents with cells from the adjacent tissue. The response of human umbilical venous smooth muscle cells (SMCs) to an excess of ferrous ions was investigated in a cell culture model at the phenotypic and at the molecular level. When soluble ferrous ions were added to the cell culture medium the cell growth rate was reduced. Gene expression profiling indicated a reduction in the amounts of mRNA from genes that are required for cell proliferation. In addition, mRNA was regulated from multiple genes involved in iron homeostasis, DNA replication and lipid metabolism. In conclusion, ions released from iron stents could reduce the vascular SMC proliferation rate by influencing growth-related gene expression and may therefore play a beneficial role in antagonizing restenosis in vivo. PMID- 16310851 TI - Hypertrophic osteoarthropathy and intrathoracic Hodgkin's disease in children. AB - BACKGROUND: Hypertrophic osteoarthropathy (HOA) is a syndrome characterized by clubbing of the fingers and toes, periosteal new bone formation of the long bones and polyarthritis. CASE REPORT: In this report, two children with intrathoracic Hodgkin's disease and HOA are presented. CONCLUSIONS: Intrathoracic neoplasms are one of the major causes of HOA in adults; however HOA is rarely associated with intrathoracic malignancies in children. HOA associated with intrathoracic Hodgkin's disease is even more rare, but should be kept in mind. PMID- 16310852 TI - Life cycle assessment of municipal solid waste management methods: Ankara case study. AB - Different solid waste management system scenarios were developed and compared for the Municipal Solid Waste Management System of Ankara by using the life cycle assessment (LCA) methodology. The solid waste management methods considered in the scenarios were collection and transportation of wastes, source reduction, Material Recovery Facility (MRF)/Transfer Stations (TS), incineration, anaerobic digestion and landfilling. The goal of the study was to determine the most environmentally friendly option of MSWM system for Ankara. The functional unit of the study was the amount of solid waste generated in the system area of concern, which are the districts of Ankara. The life cycle inventory analysis was carried out by IWM Model-1. The inputs and outputs of each management stage were defined and the inventory emissions calculated by the model were classified in to impact categories; non-renewable energy sources exhausting potential, final solid waste as hazardous and non-hazardous, global warming, acidification, eutrophication and human toxicity. The impacts were quantified with the weighing factors of each category to develop the environmental profiles of each scenario. In most of the categories, Source Reduction Scenario was found to be the most feasible management method, except the global warming category. The lowest contribution to GWP was calculated for the anaerobic digestion process. In the interpretation and improvement assessment stage, the results were further evaluated and recommendations were made to improve the current solid waste management system of Ankara. PMID- 16310853 TI - Recombinant idiotypic TCRbeta chain immunization in mice generates antigen specific T cell response. AB - Vaccination remains the most cost-effective means of preventing infectious diseases. Success of vaccination depends on generation of effective memory response. Understanding the mechanism of generation and maintenance of immunological memory would help in the design of rational vaccines. T lymphocytes play a central role in the generation of protective immune response against many microbial infections. A hypothesis known as relay hypothesis was earlier proposed, which explains the maintenance of immunological memory through interaction of idiotypic and anti-idiotypic lymphocytes. In the present study, we have shown that immunization with a model antigen, chicken ovalbumin specific T cell receptor beta chain (idiotypic TCR) generates TCR specific antibody and anti idiotypic T cell responses as well as ovalbumin specific T cell response. We further show that boosting of ovalbumin primed mice with ovalbumin specific idiotypic TCRbeta DNA or TCRbeta protein gives memory response for ovalbumin. This study provides experimental evidence for perpetuation of immunological memory through idiotypic network interactions. PMID- 16310854 TI - A novel 3-year longitudinal pilot study of medical students' acquisition and retention of screening eye examination skills. AB - OBJECTIVE: To assess the acquisition and retention of screening ophthalmic clinical skills over 3 years of medical school. DESIGN: Observational, longitudinal, multiple skills measures. PARTICIPANTS: All 96 students enrolled in a single graduating class at a public medical school in California. METHODS: Immediately after the second-year ophthalmic clinical skills course, all students were evaluated by their preceptors and self-rated for competence in defined skills. Follow-up assessments were done during 2 required third-year clerkships without additional formal ophthalmic clinical exposure. Three complete history and physical examination chart notes routinely submitted for course grading in third-year clerkships were selected randomly for each student by clerkship directors in family medicine and internal medicine, masked for identity, and then scored for appropriateness. Funduscopic skills were assessed objectively with a simulator in the third-year Clinical Performance Examination. During a fourth year 4-day ophthalmology clerkship, students were trained and reassessed with the same simulator. Just before graduation, a self-assessment questionnaire was administered to the entire class. MAIN OUTCOMES MEASURES: Twelve skills were assessed: ability to evaluate visual acuity (VA); pupils; extraocular muscles; confrontation fields; lids; cornea; conjunctiva/sclera; anterior chamber depth; and, funduscopically, the disc, macula, vessels, and retina. RESULTS: Faculty rated 88% to 90% of students as able to assess acuity, pupils, ductions, and fields, and 72% to 82% as able to visualize various parts of the fundus. Seventy six percent of students felt comfortable after funduscopic training. In 364 analyzed chart notes, one VA was measured, and pupils were examined in 66% of notes, ductions in 54%, and fields in 3%. Only 11% of notes documented attempted funduscopy; <2% suggested actual visualization. In the Clinical Performance Examination, 32% of students accurately described some aspect of the disc, with an improvement to 84% of 38 students retested after brief ophthalmology training in year 4. Of 54 (56%) respondents to the exit questionnaire, 59% felt comfortable visualizing some aspect of the fundus. CONCLUSIONS: There is worrisome erosion of students' acquired screening skills across the third-year clerkships. Skill reinforcement in the fourth year yielded improved performance. Attention must be directed to reinforcing basic ophthalmology skills training within medical school curricula to assure competence of graduates. PMID- 16310855 TI - Drug-resistant tuberculosis: a disease of target populations in Houston, Texas. AB - OBJECTIVES: To analyse the traditional and molecular epidemiology of drug resistant tuberculosis (DRTB) in Houston and Harris County, Texas in the setting of decreasing disease incidence. METHODS: Case-control study of 193 patients with DRTB and 1977 patients with drug-susceptible TB (DSTB) identified from a population-based surveillance, 1995-2001. RESULTS: In a multivariate logistic regression, the following risk factors were found to be predictors of having DRTB (P< or =0.05): human immunodeficiency virus (HIV) seropositivity, Hispanic ethnicity, Asian ethnicity, history of past TB; whereas being foreign born, having a history of past TB, and younger age were predictors of MDRTB. There were 15 patients who acquired drug resistance while on therapy, and they were significantly more likely than controls to be HIV-seropositive, be of Asian ethnicity, have smear-positive pulmonary disease and present with pleural effusion on chest radiograph. No difference in 6-month mortality between DRTB and DSTB cases was found. During the study period, the incidence of DRTB remained constant. CONCLUSIONS: In Houston, there is a steady, low-level, incidence of DRTB which disproportionately affects specific subpopulations, with no evidence of increased mortality at 6 months. PMID- 16310856 TI - Perilymph sampling from the cochlear apex: a reliable method to obtain higher purity perilymph samples from scala tympani. AB - Measurements of drug levels in the fluids of the inner ear are required to establish kinetic parameters and to determine the influence of specific local delivery protocols. For most substances, this requires cochlear fluids samples to be obtained for analysis. When auditory function is of primary interest, the drug level in the perilymph of scala tympani (ST) is most relevant, since drug in this scala has ready access to the auditory sensory cells. In many prior studies, ST perilymph samples have been obtained from the basal turn, either by aspiration through the round window membrane (RWM) or through an opening in the bony wall. A number of studies have demonstrated that such samples are likely to be contaminated with cerebrospinal fluid (CSF). CSF enters the basal turn of ST through the cochlear aqueduct when the bony capsule is perforated or when fluid is aspirated. The degree of sample contamination has, however, not been widely appreciated. Recent studies have shown that perilymph samples taken through the round window membrane are highly contaminated with CSF, with samples greater than 2microL in volume containing more CSF than perilymph. In spite of this knowledge, many groups continue to sample from the base of the cochlea, as it is a well established method. We have developed an alternative, technically simple method to increase the proportion of ST perilymph in a fluid sample. The sample is taken from the apex of the cochlea, a site that is distant from the cochlear aqueduct. A previous problem with sampling through a perforation in the bone was that the native perilymph rapidly leaked out driven by CSF pressure and was lost to the middle ear space. We therefore developed a procedure to collect all the fluid that emerged from the perforated apex after perforation. We evaluated the method using a marker ion trimethylphenylammonium (TMPA). TMPA was applied to the perilymph of guinea pigs either by RW irrigation or by microinjection into the apical turn. The TMPA concentration of the fluid sample was compared with that measured in perilymph prior to taking the sample using a TMPA-selective microelectrode sealed into ST. Data were interpreted with a finite element model of the cochlear fluids that was used to simulate each aspect of the experiment. The correction of sample concentration back to the perilymph concentration prior to sampling can be performed based on the known ST volume (4.7microL in the guinea pig) and the sample volume. A more precise correction requires some knowledge of the profile of drug distribution along the cochlear prior to sampling. This method of sampling from the apex is technically simple and provides a larger sample volume with a greater proportion of perilymph compared to sampling through the RW. PMID- 16310857 TI - Interleukin-11, IL-11 receptoralpha and leukemia inhibitory factor are dysregulated in endometrium of infertile women with endometriosis during the implantation window. AB - Interleukin (IL)-11 is essential for embryo implantation in the mouse and evidence suggests it has a role in implantation in humans. This study has evaluated immunoreactive IL-11, IL-11 receptor (R) alpha and leukemia inhibitory factor (LIF) in endometrium of infertile women with endometriosis (I/E) and normal fertile women (controls) during the implantation window. Endometrial biopsies from I/E (N = 7) were timed from the LH surge and were post-ovulatory days (POD) 5-10. Control biopsies (N = 8) from women were between days 19 and 24 of the menstrual cycle. Staining intensity of IL-11, IL-11Ralpha and LIF evaluated using semi-quantitative immunohistochemistry scores. Immunoreactive IL 11, IL-11Ralpha and LIF were present predominantly in glandular epithelium, while luminal epithelium showed patchy staining. All controls stained positively for IL 11, IL-11Ralpha and LIF in glandular epithelium. IL-11 and IL-11Ralpha staining was absent from glandular epithelium in cohorts of I/E. LIF staining intensity in glandular epithelium was significantly lower in I/E compared to controls. The results suggest that reduced endometrial IL-11 and/or LIF may contribute to infertility in some endometriotic women. PMID- 16310858 TI - Immunopathological effects of porcine circovirus type 2 (PCV2) on swine alveolar macrophages by in vitro inoculation. AB - Porcine circovirus type 2 (PCV2) is the primary causative agent of postweaning multisystemic wasting syndrome (PMWS), a multifactorial disease, in pigs. Monocyte/macrophage lineage cells, including alveolar macrophages (AMs), are the major target cells for PCV2. Swine AMs are essential for the pulmonary defense system against various pathogens. Concurrent infection of lung with opportunistic pathogens in pigs suffered from PMWS is speculated as a feature of immunosuppression. The present study was conducted to characterize the effects of PCV2 inoculation on swine AMs in the in vitro system. The parameters selected for evaluation included PCV2 antigen- and nucleic acid-containing rate, viability, TUNEL-positive rate, phagocytosis, microbicidal capability, and capacity for production of reactive oxygen species (superoxide anion, O2-, and hydrogen peroxide, H2O2), cytokines, and chemokines. High intracytoplasmic PCV2 antigen- and nucleic acid-containing rate, absence of intranuclear signals for PCV2 antigen and nucleic acid, and lack of noticeable cell death were seen in PCV2 inoculated AMs. The PCV2-inoculated AMs displayed a transient as well as persistent reduction in the up-take and destruction of Candida albicans, respectively, accompanied by decrease in the production of O2- and H2O2. In PCV2 inoculated AMs, the levels of tumor necrosis-alpha (TNF-alpha) and interleukin-8 (IL-8) were significantly increased; the mRNA expression levels of alveolar macrophage-derived neutrophil chemotactic factors-II (AMCF-II), granulocyte colony-stimulating factor (G-CSF), monocyte chemotactic protein-1 (MCP-1), and IL 8 were strongly up-regulated. The reduced phagocytosis and microbicidal capability in conjunction with decreased production of reactive oxygen species in PCV2-inoculated AMs suggest that PCV2-containing AMs may favor the survival and spread of PCV2. It is speculated that the functional alterations observed in PCV2 containing AMs may be potentially harmful to the lung tissue and local pulmonary defense system, especially in those PCV2-infected pigs conditioned by various PMWS development-dependent co-factors. PMID- 16310859 TI - Replication of West Nile virus in equine peripheral blood mononuclear cells. AB - A cell model of primary monocytes and other mononuclear cells isolated from equine blood was used to study the kinetics of West Nile virus (WNV) replication in a natural host. West Nile virus has emerged on the North American continent as a significant cause of morbidity and mortality in a wide range of avian and mammalian species. While other flaviviruses are known to infect monocytes and lymphocytes, the ability of WNV to productively replicate in specific immune cells of peripheral blood has not been assessed. In this study, enriched populations of monocytes and lymphocytes as well as purified monocytes, CD4+, CD8+ and B lymphocytes were obtained from equine blood. Productive WNV replication was demonstrated by viral growth curves, quantitative RT-PCR for WNV RNA, and indirect immunofluorescence detection of a non-structural WNV protein. Enriched and purified monocytes consistently supported productive viral replication in blood from nine of nine horses tested while a minor subset of CD4+ lymphocytes supported productive replication in cells from three of the nine horses tested. Peak viral titers of 3.2-6.6 log10 PFU/ml were reached at 6 days post-inoculation (p.i.) and titers were maintained through 10-15 days p.i. Activation of monocytes with bacterial lipopolysaccharide, which resulted in activation of nuclear transcription factor kappaB (NF-kappaB) plus elevation of nitric oxide and type I interferon levels, reduced or eliminated WNV replication. These results suggest that immune cells of the peripheral blood may serve as target cells for initial replication of WNV and may play a role in subsequent viral dissemination. Furthermore, primary equine immune cell cultures represent a potentially useful model of a natural WNV host when testing compounds such as antivirals for use in WNV treatment. PMID- 16310860 TI - Metal-dependent hydrolysis of myelin basic protein by IgGs from the sera of patients with multiple sclerosis. AB - Homogeneous IgG fractions were obtained by chromatography of the sera of patients with multiple sclerosis (MS) on Protein G-Sepharose under conditions that remove non-specifically bound proteins. These IgGs contained several chelated metals, the relative amount of which decreases in the order: Fe>or=Ca>Cu>or=Zn>or=Mg>or=Mn>or=Pb>or=Co>or=Ni. In contrast to homogeneous IgGs of healthy individuals, Abs of MS patients effectively hydrolyzed human myelin basic protein (MBP). A minor metal-dependent fraction was obtained by chromatography of highly purified IgGs from MS patient on Chelex-100. This IgG fraction did not hydrolyze human MBP in the absence of Me(2+) ions but was activated after addition of Me(2+) ions: Mg(2+)>Mn(2+)>Cu(2+)>Ca(2+). Proteolytic activities of IgGs from other MS patients were also activated by other metal ions (Ni(2+), Fe(2+), Co(2+), Zn(2+), Pb(2+), and Co(2+)) and especially Ni(2+). Ni(2+)-activated IgGs were separated into distinct MBP-hydrolyzing fractions by chromatography on HiTraptrade mark Chelating Sepharose charged with Ni(2+). Detection of Mg(2+)-dependent proteolytic activity in the SDS-PAGE area corresponding only to IgG provided direct evidence that IgG from sera of MS patients possesses metal-dependent human MBP-hydrolyzing activity. Observed properties of MS abzymes distinguish them from other known mammalian metalloproteases and demonstrate their pronounced catalytic diversity. Metal dependent IgGs from MS patients represent the first example of abzymes with metal dependent proteolytic activity. PMID- 16310861 TI - Positioning prostanoids of the D and J series in the immunopathogenic scheme. AB - Prostaglandin D(2) (PGD(2)) is produced by a variety of immune and non hematopoietic cells and appears to function in both an inflammatory and homeostatic capacity. Two genetically distinct PGD(2)-synthesizing enzymes have been identified to date, including hematopoietic- and lipocalin-type PGD synthases (H-PGDS and L-PGDS, respectively). Though the inter-species expression profiles of these two enzymes vary widely, H-PGDS is generally localized to the cytosolic aspect of immune and inflammatory cells, whereas L-PGDS is more resigned to tissue-based expression. PGD(2) activity is principally mediated through two unique G protein-coupled receptors (GPCR), designated DP(1) and DP(2). These receptors exhibit overlapping binding profiles, yet their respective agonists elicit generally distinctive responses. Additional to DP receptors, the PGD(2) metabolite 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)) binds the nuclear peroxisome proliferator-activated receptor gamma (PPARgamma) and has the facility to initiate a variety of anti-inflammatory phenotypes either through or independent of PPARgamma association. This review highlights the collective relevance of PGD(2) and its respective synthases, receptors, and metabolites in immunopathologic responses. PMID- 16310862 TI - UK and Ireland experience of bone anchored hearing aids (BAHA) in individuals with Down syndrome. AB - OBJECTIVE: To evaluate the indications, surgical techniques and post-operative problems seen in children with Down syndrome fitted with bone anchored hearing aids (BAHA). METHODS: A postal survey of all registered United Kingdom and Ireland BAHA centres. RESULTS: There was a 98% response rate to the survey. Eighty-one centres in the United Kingdom and Ireland undertake BAHA surgery. Eighteen centres were identified to have undertaken BAHA surgery on individuals with Down syndrome. A total of 43 individuals were implanted. Twenty-four cases were under the age of 16. Soft tissue complications were encountered in 21 patients (49%). Osseointegration failures were encountered in four cases (9%). There was a high level of satisfaction with the BAHA system amongst patients, parents and carers. CONCLUSIONS: BAHA is a valuable method of hearing amplification in children with Down syndrome. It should be considered not as a primary method of amplification, but in the overall management of individuals with Down syndrome after conventional hearing aids and/or ventilation tubes have been considered or already failed. The survey shows a high patient and carer satisfaction with the system, despite short term early soft tissue complications. PMID- 16310863 TI - High frequency of Alloiococcus otitidis in the nasopharynx and in the middle ear cavity of otitis-prone children. AB - OBJECTIVE: Colonization of middle ear pathogens is facilitated in the nasopharynx of otitis-prone children, and is associated with the development of otitis media. Recently, a new species of bacterium, Alloiococcus otitidis, is considered as one of the major middle ear pathogens. However, as far as we know, no study has been reported concerning the prevalence of A. otitidis in the nasopharynx of otitis prone children. And, no study has been conducted on the association of A. otitidis in the nasopharynx with otitis media. METHODS: The frequency of A. otitidis in 83 middle ear effusions (MEE) and 56 nasopharyngeal swab (NPS) specimens from 56 children with otitis media was investigated by culture and polymerase chain reaction (PCR). RESULTS: A. otitidis was detected in 24 (28.9%) of MEE and in 6 (10.7%) of NPS specimens. When the frequency was investigated in relation to proneness to otitis media, A. otitidis was detected in 16 (64%) of 25 MEE and in 5 (29.4%) of 17 NPS specimens from otitis-prone children, whereas it was detected in 8 (13.8%) of 58 MEE and in 1 (2.6%) of 39 NPS specimens from non otitis-prone children. The frequency of A. otitidis in both NPS and MEE specimens was significantly higher in otitis-prone than in non-otitis-prone children. CONCLUSIONS: Our results suggest that colonization of A. otitidis is facilitated in the nasopharynx of otitis-prone children. And, nasopharyngeal colonization of A. otitidis may be associated with the frequency of otitis media. PMID- 16310864 TI - Chemokine blockers--therapeutics in the making? AB - Chemokines are a family of small chemoattractant cytokines that have an important role in controlling leukocyte migration. The finding that some chemokines and their receptors are upregulated in both acute and chronic inflammatory diseases, and that they are key players in the development of AIDS, has provided the pharmaceutical industry with new targets for therapeutic intervention in these diseases. Although the chemokine system shows apparent redundancy in vitro, target validation is possible largely through expression studies in human disease tissues and the use of transgenic and knockout mice as disease models. Several approaches are being developed to block the effects of chemokines, including small-molecule antagonists of chemokine receptors, modified chemokines and antibodies directed against chemokine receptors. Here, we describe the rationale behind these different approaches, the pitfalls that have been encountered and future perspectives. PMID- 16310865 TI - The expression and function of chemokines involved in CNS inflammation. AB - Chemokines and their receptors have principal roles in leukocyte trafficking under normal physiological and pathological conditions. The differential expression of the chemokine system in different parts of the CNS provides insights into the processes that are required for normal immune surveillance and pathological immune-mediated effector processes. Insights derived from studying multiple sclerosis, an inflammatory disorder of the CNS in humans, and experimental autoimmune encephalomyelitis, an animal model of this disorder, aid in further understanding the complexities of chemokine-mediated inflammation. Knowledge of the molecular biology of chemokines and their receptors, and the roles of specific chemokine ligands and receptors in the CNS in health and in disease have made these proteins targets for therapeutic intervention in neuroinflammation. We also discuss currently proposed and potentially useful chemokine receptor antagonists. PMID- 16310866 TI - Electron microscopic examination of the viromatrix of Rana grylio virus in a fish cell line. AB - Rana grylio virus (RGV), a Ranavirus belonging to the family Iridoviridae, assembles in the viromatrix which is a factory for viral genome replication and particle assembly. Ultrastructural studies of the viromatrix will clarify the pathway of assembly. The viromatrix and quantitative changes in RGV infected epithelipma papulosum cyprini (EPC) cells, one of fish cell lines, were studied by electron microscopy. It was shown that viromatrices were adjacent to the nucleus, and the electron density was lower than that of the surrounding cytoplasm. The viromatrix contained virus particles with different forms, electron-dense materials and amorphous structures which included tubules and membranous materials. Tubules were often observed in direct continuity with empty capsids. Several bundles of intermediate filaments were seen alongside the viromatrix and crystalline aggregates. Large clusters of mitochondria occurred in proximity to viromatrix. A total of 990 cells profiles were examined. The results showed that 394 cells contained viromatrix: 89.3% contained one, and 10.7% contained two to four viromatrices. The number of viromatrices increased gradually and reached a peak at 16 h p.i. The viromatrix area at 24 h p.i. increased up to 7.4 +/- 0.69 microm(2) which was three-times lower than that at 6 h p.i. The number of empty capsids within viromatrix was generally more than that of "full" particles at different time points, and there was a strong positive correlation between them. PMID- 16310867 TI - Quantitative detection of a marine fish iridovirus isolated from large yellow croaker, Pseudosciaena crocea, using a molecular beacon. AB - A real-time polymerase chain reaction (PCR) assay utilizing a molecular beacon for the quantitative detection of a marine fish iridovirus isolated from large yellow croaker, Pseudosciaena crocea (LYCIV), was developed, which involved the amplification of a 122bp DNA fragment from a conserved region of LYCIV ATPase gene. The specific probe consisting of two short arm and a central loop sequences complementary to the target amplicon was characterized with respect to its efficiency of quenching (E(ff)), and signal to background ratio by spectrofluorometric analysis of its hybridization with the complementary oligonucleotide target. The positive control plasmid pFHT-ATPase containing the target sequence was quantified to make the standard curve for sample detection after serial 10-fold dilution. Linear coefficient correlations between cycle threshold (C(T)) value and logarithmic positive plasmid concentration were close to one (r(2)=0.998) and the detection limit of the assay was 70 copies of positive plasmid/assay. The specificity of this real-time PCR was also demonstrated by using the genomic DNA templates from the healthy fish, white spot syndrome baculovirus (WSSV), and epizootic heamatopietic necrosis virus (EHNV), respectively. The coefficient of variation (CV) of the assay ranged from 1.16 to 4.42%, depending on the concentration of the positive plasmid. The quantitative detection of different tissues from LYCIV-infected fish showed that the spleen and kidney contained the largest number of viral particles (6.86 x 10(6) and 4.62 x 10(6) viral genome copies/mg tissue, respectively) while no viral DNA was detected in the muscular tissue. These results suggested that the real-time PCR assay reported here could be used for rapid, sensitive, and quantitative detection of LYCIV infection. PMID- 16310868 TI - Avian metapneumovirus phosphoprotein targeted RNA interference silences the expression of viral proteins and inhibits virus replication. AB - Avian metapneumovirus (aMPV) is one of the major causes of serious respiratory infections of poultry and leads to considerable economic losses to food animal production worldwide. Here, we show that double stranded short interfering RNA (siRNA) molecules corresponding to aMPV phosphoprotein (P) gene silence P RNA and protein expression. These siRNAs broadly reduced the expression of other viral proteins in addition to P, but did not have a discernable effect on cellular protein expression. The exposure of cells to P-specific siRNAs also led to inhibition of virus replication as evidenced by marked reduction in the progeny virion titers. Taken together, the findings suggest that exogenous P silencing siRNAs can inhibit aMPV replication with potential implications in the design of novel siRNA based prophylactics. PMID- 16310869 TI - C-section birth per se or followed by acute global asphyxia altered emotional behaviour in neonate and adult rats. AB - Birth complications such as perinatal asphyxia are considered risk factors for later neurobehavioural disorders. Behavioural analysis of animal models may help to clarify the contribution of particular patterns of early hypoxia and their combination to psychiatric morbidity. Wistar rats underwent caesarean section (c section) alone or c-section followed by asphyxia, the latter induced by placing pups still in uterus horns into a water bath at 37 degrees C for 20 min. Vaginally delivered pups were used as controls. Frequency of ultrasound emissions was analysed following isolation at a lower temperature than that of the home nest (23+/-0.5 degrees C) and reunion with their mother (3 min) on postnatal day (PND) 13 (maternal potentiation test). A sex-dependent effect of hypoxia was observed, with higher production of ultrasounds in hypoxic males. Caesarean delivered pups produced significantly more ultrasounds than those vaginally delivered. At adolescence (PND 35) rats underwent a 25 min social interaction test with a conspecific of the same sex and age. Significant alterations in investigative behaviour (inclusive of: nose, anogenital, body sniffing, and following) were evident in caesarean-delivered rats of both sexes, but not in rats experiencing perinatal asphyxia. At adulthood, auditory, and context conditioned responses, analysed in a fear conditioning test, were not markedly affected either by c-section or c-section plus hypoxia. However, hypoxic rats emitted significantly more 22 kHz ultrasounds than c-section or vaginally delivered rats during the training session. In conclusion, differential effects appear to be brought about by c-section and by hypoxia mainly related to emotional/anxious responses. PMID- 16310870 TI - Anxiety and cognition in female histidine decarboxylase knockout (Hdc(-/-)) mice. AB - The role of histamine in brain function has been studied using histidine decarboxylase (HDC) deficient male mice. As the effects of HDC deficiency on brain function might be sex-dependent, we behaviorally analyzed Hdc(-/-) and control female mice. Compared to female control mice, Hdc(-/-) female mice showed hypoactivity, increased measures of anxiety, impairments in water-maze performance, but enhanced passive avoidance memory retention. Following behavioral testing, arginine vasopression (AVP) immunoreactivity was higher in the dorsal hypothalamus and central and basolateral nuclei of the amygdala of Hdc(-/-) than Hdc(+/+) mice. Finally, MAP2 immunoreactivity in the hippocampal CA1 region correlated positively with measures of anxiety in the open-field and light-dark tests and negatively with performance during the hidden sessions of the water-maze. As the effects of HDC deficiency on object recognition, water maze, and rotorod performance, were sex-dependent, it is important to consider potential effects of sex in the interpretation of the role of histaminergic neurotransmission in brain function. PMID- 16310871 TI - Localized lesions of ventral striatum, but not arcopallium, enhanced impulsiveness in choices based on anticipated spatial proximity of food rewards in domestic chicks. AB - The effects of bilateral chemical lesions of the ventral striatum (nucleus accumbens and the surrounding areas in the medial striatum) and arcopallium (major descending area of the avian telencephalon) were examined in 1-2-weeks old domestic chicks. Using a Y-maze, we analyzed the lesion effects on the choices that subject chicks made in two tasks with identical economical consequences, i.e., a small-and-close food reward vs. a large-and-distant food reward. In task 1, red, yellow, and green beads were associated with a feeder placed at various distances from the chicks; chicks thus anticipated the spatial proximity of food by the bead's color, whereas the quantity of the food was fixed. In task 2, red and yellow flags on the feeders were associated with various amount of food; the chicks thus anticipated the quantity of food by the flag's color, whereas the proximity of the reward could be directly visually determined. In task 1, bilateral lesions of the ventral striatum (but not the arcopallium) enhanced the impulsiveness of the chicks' choices, suggesting that choices based on the anticipated proximity were selectively changed. In task 2, similar lesions of the ventral striatum did not change choices. In both experiments, motor functions of the chicks remained unchanged, suggesting that the lesions did not affect the foraging efficiency, i.e., objective values of food. Neural correlates of anticipated food rewards in the ventral striatum (but not those in the arcopallium) could allow chicks to invest appropriate amount of work-cost in approaching distant food resources. PMID- 16310872 TI - Application and validation of approaches for the predictive hazard assessment of realistic pesticide mixtures. AB - In freshwater systems located in agricultural areas, organisms are exposed to a multitude of toxicologically and structurally different pesticides. For regulatory purposes it is of major importance whether the combined hazard of these substances can be predictively assessed from the single substance toxicity. For artificially designed multi-component mixtures, it has been shown that the mixture toxicity can be predicted by concentration addition (CA) in case of similarly acting substances and by independent action (IA), if mixtures are composed of dissimilarly acting substances. This study aimed to analyse whether these concepts may also be used to predictively assess the toxicity of environmentally realistic mixtures. For this purpose a mixture of 25 pesticides, which reflects a realistic exposure scenario in field run-off water, was studied for its effects on the reproduction of the freshwater alga Scenedesmus vacuolatus. The toxicity of the tested mixtures showed a good predictability by CA. This is consistent with the finding that the toxicity was dominated by a group of similarly acting photosystem II inhibitors, although the mixture included substances with diverse and partly unknown mechanisms of action. IA slightly underestimated the actual mixture toxicity. However, the EC(50) values that can be derived from each prediction, according to CA respectively IA, only differed by a factor of 1.3. The finding of such a small difference is partly explainable by the fact that only few components dominate the mixture scenario in terms of so-called toxic units (TUs). This connection is established by developing an equation that allows to calculate the maximum possible ratio between corresponding predictions of effect concentrations by IA and CA for any given ratio of the TUs of mixture components, irrespective of their individual concentration-response functions and independent from their mechanisms of action. To evaluate whether small quantitative differences between EC(50) values predicted by CA and IA are an exception or rather the rule for agricultural exposure scenarios, this calculation was applied on an additional set of 18 pesticide exposure scenarios that were taken from the literature. For these scenarios, EC(50) values predicted by IA can never exceed those predicted by CA by more than a factor of 2.5. The findings of this study support the view that CA provides a precautious but not overprotective approach to the predictive hazard assessment of pesticide mixtures under realistic exposure scenarios, irrespective of the similarity or dissimilarity of their mechanisms of action. PMID- 16310873 TI - Biphasic decay of guide RNAs in Trypanosoma brucei. AB - Guide RNAs (gRNAs) are short mitochondrially encoded RNAs that contain the information for editing of messenger RNAs in Trypanosoma brucei. Although a great deal of work has focused on the utilization of gRNAs in editing, little is known about the turnover of gRNAs. In this report, we utilized in organello pulse chase and in vitro RNA decay experiments to directly examine gRNA turnover. We found that gRNAs are degraded by a biphasic mechanism. In the first step of decay, 3' gRNA sequences encompassing primarily the post-transcriptionally added oligo(U) tail are rapidly removed. This is followed by a second step, which entails a comparatively slower degradation of the encoded gRNA body. Decay of the 3' end of the gRNA is sequence specific, as it does not occur on oligoadenylated gRNAs. In contrast, the nucleotide composition of the 3' extension does not affect the rate of degradation during the second, slower, decay step. Finally, competition assays suggest that complete gRNA decay is mediated by two distinct enzymes, one of which simultaneously recognizes elements of the oligo(U) tail and the encoded portion of the gRNA. Overall, these results provide the first evidence for a gRNA specific decay pathway. PMID- 16310874 TI - Cardioverter defibrillator implantation in an adult with isolated noncompaction of the ventricular myocardium. AB - Isolated noncompaction of the ventricular myocardium is an unclassified cardiomyopathy and is thought to be due to arrest of myocardial morphogenesis. Fatal ventricular arrhythmias may occur in approximately half of the patients and account for half of the death in this disorder. In this report, we describe a patient with isolated noncompaction of the ventricular myocardium in whom implantation of automatic cardioverter defibrillator was thought to be effective to prevent the risk of sudden cardiac death. PMID- 16310875 TI - Analysis of covariance patterns in gene expression data and FT-IR spectra. AB - The aim of this study was to detect and interpret correlation patterns in several large data matrices from the same biological system using Partial Least Squares Regression (PLSR) in order to get information on the system under investigation. To do this, DNA microarray data and Fourier Transform Infrared (FT-IR) spectra from a designed study where Campylobacter jejuni was exposed to environmental stress conditions, were used. The experimental design included variation in atmospheric conditions, temperature and time. PLSR was first used to analyse each of the two data types separately in order to explore the effect of the experimental parameters on the data. The results showed that both the gene expression and FT-IR spectra were affected by the variations in atmosphere, temperature and time, but that the effect was different for the two types of data. When the DNA microarray data and FT-IR spectra were linked together by PLSR, covariation due to temperature was seen. Both specific genes and ranges in the FT-IR spectra that were connected to the variation in temperature were detected. Some of these are possibly connected to properties of the cell wall of the bacteria. The results in this study show the potential of PLSR for investigation of covariance structures in biological data. By doing this, valuable information about the biological system can be detected and interpreted. It was also shown that the use of FT-IR spectroscopy provided important information about the stress responses in the bacteria, information that was not detected from the DNA microarray data. PMID- 16310876 TI - G protein-activated inwardly rectifying K+ channel inhibition and rescue of weaver mouse motor functions by antidepressants. AB - Antidepressants, including tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs), have been widely used for the treatment of not only depression but also other psychiatric disorders, although the molecular mechanisms of the drug effects have not yet been sufficiently revealed. Here, we investigated the in vivo effects of these antidepressants on G protein-activated inwardly rectifying K+ (GIRK) channels, which are important for regulating the excitability of various cells, by using weaver (wv) mice, which have mutant GIRK channels and show abnormal neuronal cell death and motor disturbances. First, we found that a widely used SSRI fluoxetine (also known as Prozac) effectively inhibited wv GIRK2 channels like wild-type GIRK channels, expressed in Xenopus oocytes. Next, we found that weaver motor disturbances were remarkably alleviated by chronic treatment with fluoxetine or desipramine. Furthermore, the chronic fluoxetine treatment substantially suppressed the abnormal neuronal cell death in the weaver mouse cerebellum and pontine nuclei. These results suggest that continuous inhibition of wv GIRK2 channels by a group of antidepressants caused substantial suppression of the neuronal cell death and resulted in improvement of motor abilities in weaver mice. These results provide evidence for in vivo GIRK channel inhibition by a group of antidepressants. PMID- 16310877 TI - Functional specialization within the dorsolateral prefrontal cortex: a review of anatomical and physiological studies of non-human primates. AB - The dorsolateral prefrontal cortex (DLPFC) possesses cortico-cortical connections with the parietal and premotor cortices that are involved in visuomotor control of actions. Studies have shown that the DLPFC, especially the caudal part, has a crucial role in cognitive control of motor behavior, and that it uses spatial information in conjunction with information such as object identity, behavioral rules, and rewards. Current anatomical and physiological studies indicate that the DLPFC may not be a single entity. Anatomical studies show that preferential anatomical connections exist between subregions of the DLPFC and the parietal/premotor cortices. Physiological studies based on data obtained from monkeys performing a variety of cognitive tasks report region-specific neuronal activity within the DLPFC. In this article, I review evidence for functional segregation within the DLPFC and postulate that at least two distinct subregions, i.e., the dorsal and ventral parts, can be identified. PMID- 16310878 TI - Through central arginine vasopressin, not oxytocin and endogenous opiate peptides, glutamate sodium induces hypothalamic paraventricular nucleus enhancing acupuncture analgesia in the rat. AB - Our previous study proved that the hypothalamic paraventricular nucleus (PVH) plays an important role in acupuncture analgesia. The neuropeptides involving in the PVH regulation of acupuncture analgesia was investigated in the rat. The changes of pain threshold, which was induced by electrical acupuncture of "Zusanli" points (St. 36), were measured as acupuncture analgesia. Microinjection of l-glutamate sodium into the PVH, which only excites the PVH neurons, could dose-dependently enhance the acupuncture analgesia, but microinjection of l glutamate sodium into the area nearby the PVH did not alter acupuncture analgesia. Removing pituitary did not influence this effect of l-glutamate sodium. Microinjection of l-glutamate sodium into the PVH only increased the arginine vasopressin (AVP), not oxytocin (OXT), leucine enkephaline (L-Ek), beta endorphine (beta-Ep) and dynorphinA(1-13) (DynA(1-13)) concentrations in the PVH perfuse liquid using radioimmunoassay. Intraventricular injection of anti arginine vasopressin serum (AAVPS) could completely reverse the effect of microinjection of l-glutamate sodium into the PVH enhancing acupuncture analgesia. Intraventricular injection of naloxone, one opiate peptide antagonist, partly attenuated this effect of l-glutamate sodium, and intraventricular of anti oxytocin serum (AOXTS) did not change this effect of l-glutamate sodium. The results suggested that l-glutamate sodium induces the PVH enhancing acupuncture analgesia only through AVP, not OXT and endogenous opiate peptides in central nervous system. PMID- 16310879 TI - Multiclonal pattern of Jaagsiekte sheep retrovirus integration sites in ovine pulmonary adenocarcinoma. AB - Insertional mutagenesis and envelope (Env)-mediated oncogenesis are hypothesized mechanisms by which Jaagsiekte sheep retrovirus (JSRV) causes ovine pulmonary adenocarcinoma (OPA). Twenty-eight JSRV integration sites in lung tumors (LTs) from four sheep with OPA were cloned and sequenced by a multiple step gene walking technique. Using nested PCR, clonal expansion of these integration sites could be detected, if at all, only in the localized regions of LT from which the integration sites were derived. One sheep had a viral integration site in a sequence with 85 and 81% identity, respectively, over 100 bp to exon 2 of the human and mouse receptor protein tyrosine phosphatase gamma genes. Clonal integration of Jaagsiekte sheep retrovirus in this gene was demonstrated by nested PCR and Southern blot hybridization in the DNA sample from which the integration site was cloned, but not in other LT or kidney DNA samples from the same sheep. OPA may develop from multiple independent oncogenic events and a role for insertional mutagenesis cannot be ruled out. PMID- 16310880 TI - Programmed ribosomal frameshifting in HIV-1 and the SARS-CoV. AB - Ribosomal frameshifting is a mechanism of gene expression used by several RNA viruses to express replicase enzymes. This article focuses on frameshifting in two human pathogens, the retrovirus human immunodeficiency virus type 1 (HIV-1) and the coronavirus responsible for severe acute respiratory syndrome (SARS). The nature of the frameshift signals of HIV-1 and the SARS-CoV will be described and the impact of this knowledge on models of frameshifting will be considered. The role of frameshifting in the replication cycle of the two pathogens and potential antiviral therapies targeting frameshifting will also be discussed. PMID- 16310881 TI - The threshold value for insulin resistance (HOMA-IR) in an admixtured population IR in the Brazilian Metabolic Syndrome Study. PMID- 16310882 TI - Salvage effect of the vascular endothelial growth factor on chemically induced acute severe liver injury in rats. AB - BACKGROUND/AIMS: The role of the vascular endothelial growth factor (VEGF), a potent angiogenic factor, in liver regeneration following acute severe liver injury (ALI) has not been elucidated. The aims of the current study were to investigate the role of VEGF, and to find out whether VEGF can improve the outcome of ALI in rats. METHODS: ALI was induced in male rats by combination of D galactosamine (Gal-N) and lipopolysaccharide (LPS). The survival rate and several indices were chronologically compared with or without VEGF treatment. RESULTS: The overall survival rate of the VEGF-treated group significantly improved as compared with the untreated group (100 vs. 27%, respectively). The serum ALT elevation, with a peak at 24 h after Gal-N+LPS intoxication, was markedly attenuated with VEGF treatment. The proliferation of hepatocytes and sinusoidal endothelial cells (SEC) was stimulated by VEGF with a peak at 36 and 96 h, respectively. The immunohistochemical analysis revealed that VEGF drastically prevented destruction of the SEC architecture in ALI. Our in vitro study showed that VEGF significantly prevented the Gal-N+LPS-induced cytotoxicity and apoptosis of SEC. CONCLUSIONS: VEGF treatment significantly reduced the mortality rate of ALI in the rat, and it may provide a new therapeutic strategy for ALI. PMID- 16310883 TI - Superoxide anions and hydrogen peroxide induce hepatocyte death by different mechanisms: involvement of JNK and ERK MAP kinases. AB - BACKGROUND/AIMS: In liver diseases, reactive oxygen species (ROS) are involved in cell death and liver injury, but the mechanisms are not completely elucidated. To elucidate the mechanisms of hepatocyte cell death induced by the ROS superoxide anions and hydrogen peroxide, primary cultures of hepatocytes were exposed to the superoxide anion donor menadione (10-50 micromol/L) or H2O2 (1-5 mmol/L). Hepatocytes were also treated with caspases and MAPKs inhibitors, superoxide dismutase (PEG-SOD) and SNAP, a nitric oxide donor. Apoptosis was determined by measuring caspase-9, -6, -3 activation and cleaved PARP, and necrotic cell death by Sytox Green staining. RESULTS: (1) Menadione (50 micromol/L) induces JNK phosphorylation, caspase-9, -6, -3 activation, PARP cleavage and apoptosis. Superoxide anions-induced apoptosis is dependent on JNK activity. Menadione (50 micromol/L) induces the phosphorylation of ERK1/2 and this attenuates cell death. (2) H2O2 increases necrotic cell death at high concentration or when H2O2 detoxification is impaired. H2O2 does not activate MAPKs signalling. (3) PEG-SOD prevents ERK1/2-, JNK- phosphorylation, caspase activation and apoptosis induced by menadione. Glutathione depletion increases menadione-induced apoptosis. (4) SNAP abolishes menadione-induced apoptosis but increases necrotic cell death. CONCLUSIONS: In normal hepatocytes, superoxide anions-induced caspase activation and apoptosis is dependent on JNK activity and totally abolished by superoxide scavengers. PMID- 16310884 TI - Recent progress in ocular drug delivery for posterior segment disease: emphasis on transscleral iontophoresis. AB - Age-related macular degeneration, diabetic retinopathy, posterior uveitis, and retinitis due to glaucoma are leading causes of vision loss in the United States and other developed countries. Because these diseases are located in the posterior segment of the eye, topical application of ophthalmic medicines is of limited benefit, since topically applied drugs rarely reach therapeutic levels in the affected posterior tissues such as the choroid and retina. Intravitreal injections can deliver drugs to the posterior segment without the side effects associated with systemic administration. However, the repeated and long-term injections often needed may cause complications, such as vitreous hemorrhage, retinal detachment, or endophthalmitis. Recent advances in ocular drug delivery methods and the development of novel biopharmaceutical agents could lead to new regimens for the treatment of disease of the posterior retina, choroids, and macula. This review will summarize recent literature concerning ocular drug delivery of bioactive compounds to the posterior segment of the eye with emphasis on transscleral iontophoresis. PMID- 16310885 TI - Supramolecular structures from dendrons and dendrimers. AB - This paper reviews aspects of the association of dendrons and dendrimers into a variety of supramolecular structures. There is such a wide range of primary dendron and dendrimer chemistries that it is still difficult to predict behaviour in aqueous media, and there are few studies in non-aqueous media. The aggregation of the primary units into larger and more complex forms leads to a wider range of potential carrier systems for drugs, genes and vaccines. This review deals principally with the association structures which can be formed. These include liquid crystalline structures and dendron block copolymer aggregates, surface monolayer formation, dendrimer derived nanoparticles, micellar structures and dendrisome (vesicle) formation. Of particular interest are DNA-dendrimer complexes and dendrimer-polyanion interactions. The in vivo behaviour of dendrons and dendrimers is of course crucial and is addressed. Dendrimer vesicle solubilisation by surfactants and emulsion stabilisation by dendrimers completes the survey of secondary structures. The challenge is to understand better the processes involved and to concentrate further on the design of the synthesis of dendrons and dendrimers which will associate into specific complex structures to increase the scope of dendrimer science. PMID- 16310886 TI - Randomized multicentric phase II study of carboplatin/gemcitabine and cisplatin/vinorelbine in advanced non-small cell lung cancer GFPC 99-01 study (Groupe francais de pneumo-cancerologie). AB - PURPOSE: To evaluate the efficacy and safety of gemcitabine and carboplatin in the treatment of previously untreated patients with advanced non-small cell lung cancer (NSCLC). METHODS: A randomized phase II study was conducted by the Groupe Francais de Pneumo-Cancerologie (GFPC) in 15 centers. The patients were randomized in either arm A (GC): gemcitabine 1250 mg/m2 on days 1 and 8+carboplatin AUC 6 mg/(mLmin) on day 1; or in arm B (VP): vinorelbine 30 mg/m2 weekly+cisplatin 80 mg/m2 on day 1. Treatment cycles were repeated every 3 weeks. RESULTS: A total of 100 patients were randomized with stage IV or stage III NSCLC with malignant pleural effusion: 51 patients in arm A and 49 patients in arm B. A total of 190 cycles were administered in the GC arm and 172 cycles in the VP arm, with a median of four cycles per patient in each arm. The dose intensity was 84.9% for gemcitabine, 99.8% for carboplatin, 97.7% for cisplatin and 67.7% for vinorelbine. The objective response rates were 19.6% (95% CI, 9.8-33.1) for GC and 29.2% (95% CI, 17.0-44.1) for VP in an ITT analysis. The response duration was 169 days in arm A and 226 days in arm B. The TTP was similar with 140 days (GC) and 148 days (VP), respectively. Overall survival rates were 334 days in the GC combination and 304 days in the VP combination. Overall, the treatment was safe and toxicities observed were different in each arm: neutropenia was the most common toxicity in the VP treatment, whereas thrombocytopenia was more frequent in the GC combination. Anemia was similar in both arms. Non-haematologic toxicity was mild. One toxic death in arm A and three toxic deaths in arm B were observed. CONCLUSION: In terms of response rate, the gemcitabine-carboplatin combination was not efficient enough to allow further phase III study. Survival data are in the same range as the standard arm. This chemotherapy is feasible and may represent an alternative to a standard cisplatin-based regimen, allowing treatment in an outpatient setting. PMID- 16310888 TI - Hydraulic characterization for steam enhanced remediation conducted in fractured rock. AB - To explore the viability of Steam Enhanced Remediation (SER) in fractured rock a small-scale steam injection and water/vapour extraction pilot study was conducted at the former Loring Air Force Base in northern Maine, USA. A detailed well testing program was undertaken to assist in the design of the injection and extraction well array, and to assess the possibility of off-site heat and contaminant migration. A structurally complex limestone having low matrix porosity and a sparse distribution of fractures underlies the study site. To characterize the groundwater and steam flow pathways, single-well slug tests and more than 100 pulse interference tests were conducted. The results of the well testing indicate that the study site is dominated by steeply dipping bedding plane fractures that are interconnected only between some wells in the injection/extraction array. The SER system was designed to take advantage of interconnected fractures located at depth in the eastern end of the site. An array of 29 wells located in an area of 60 by 40 m was used for steam injection and water/vapour extraction. The migration of heat was monitored in several wells using thermistor arrays having a 1.5 m vertical spacing. Temperature measurements obtained during and after the 3 month steam injection period showed that heat migration generally occurred along those fracture features identified by the pulse interference testing. Based on these results, it is concluded that the pulse interference tests were valuable in assisting with the design of the injection/extraction well geometry and in predicting the migration pathways of the hot water associated with the steam injection. The pulse interference test method should also prove useful in support of any other remedial method dependant on the fracture network for delivery of remedial fluid or extraction of contaminants. PMID- 16310889 TI - Modeling and interpreting bioavailability of organic contaminant mixtures in subsurface environments. AB - Bioavailability often controls the fate of organic contaminants in surface and subsurface aquatic environments. Bioavailability can be limited by sorption, mass transfer, and intrinsic biodegradation potential and can be further altered by the presence of other compounds. This paper reviews current perspectives on the processes influencing subsurface contaminant bioavailability, how these processes are modeled, and how the relative role of the various processes can be assessed through bioavailability indices. Although these processes are increasingly well understood, the use of sophisticated models and indices often are precluded by an inability to estimate the many parameters that are associated with complex models. Nonetheless, the proper representation of sorption, mass transfer, biodegradation, and co-solute effects can be critical in predicting bio attenuation. The influence of these processes on contaminant fate is illustrated with numerical simulations for the simultaneous degradation of toluene (growth substrate) and trichloroethylene (nongrowth cometabolite) in hypothetical, aerobic, solid-water systems. The results show how the relative impacts on contaminant fate of the model's various component processes depends upon system conditions, including co-solute concentrations. Slow biodegradation rates increase the inhibition effects of a cometabolite and suppress the rate enhancement effects of a growth substrate. Irrespective of co-solute effects, contaminant fate is less sensitive to biodegradation processes in systems with strong sorption and slow desorption rates. Bioavailability indices can be used to relate these findings and to help identify appropriate modeling simplifications. In general, however, there remains a need to redefine such indices in order that bioavailability concepts can be better incorporated into site characterization, remediation design, and regulatory oversight. PMID- 16310890 TI - Guidelines for the control of glycopeptide-resistant enterococci in hospitals. AB - The increase since the mid 1980s in glycopeptide resistant enterococci (GRE) raised concerns about the limited options for antimicrobial therapy, the implications for ever-increasing numbers of immunocompromised hospitalised patients, and fuelled fears, now realised, for the transfer of glycopeptide resistance to more pathogenic bacteria, such as Staphylococcus aureus. These issues underlined the need for guidelines for the emergence and control of GRE in the hospital setting. This Hospital Infection Society (HIS) and Infection Control Nurses Association (ICNA) working party report reviews the literature relating to GRE prevention and control. It provides guidance on microbiological investigation, treatment and management, including antimicrobial prescribing and infection control measures. Evidence identified to support recommendations has been categorized. A risk assessment approach is recommended and areas for research and development identified. PMID- 16310891 TI - Rapid modulation of TRH-like peptides in rat brain by thyroid hormones. AB - Recent identification of membrane receptors for T4, T3, 3,5-T2, and 3 iodothyronamine that mediate rapid physiologic effects of thyroid hormones suggested that such receptors may supplement the regulation of TRH and TRH-like peptides by nuclear T3 receptors. For this reason 200 g male Sprague-Dawley rats received daily i.p. injections of PTU or T4. Levels of TRH and TRH-like peptides were measured 0, 2 h or 1, 2, 3, or 4 days later. Rapid increases or decreases in TRH and TRH-like peptide levels were observed in response to PTU and T4 treatments in various brain regions involved in mood regulation. Significant effects were measured within 2 h of T4 injection. Nuclear T3 receptor-mediated changes in gene expression altering translation, post-translational processing and constitutive release of peptides require more than 2 h. We conclude that non genomic mechanisms may contribute to the psychiatric effects of thyroid disease and thyroid hormone adjuvant treatment for major depression. PMID- 16310892 TI - Longitudinal changes in white matter following ischemic stroke: a three-year follow-up study. AB - Information on longitudinal changes in white matter after stroke is limited. The aim of the present study was to quantitatively investigate longitudinal changes in the microstructural integrity of non-lesioned white matter at 1-3 years following ischemic stroke. In a sample of 80 ischemic stroke patients, we obtained diffusion tensor imaging (DTI) measures of fractional anisotropy (FA), an apparent measure of white matter integrity, in radiologically normal-appearing white matter at baseline and 3 years of follow-up. Mixed model regression analysis results showed a significant improvement in FA from baseline during the first 2 years of follow-up that stabilized by the third year of follow-up. These results demonstrate a long-term improvement in apparent white matter integrity following ischemic stroke that continues, at least, into the second year following the insult. PMID- 16310893 TI - Oxidation, glycoxidation, lipoxidation, nitration, and responses to oxidative stress in the cerebral cortex in Creutzfeldt-Jakob disease. AB - Gel electrophoresis and Western blotting of frontal cortex homogenates have been carried out in sporadic Creutzfeldt-Jakob disease (CJD) cases and age-matched controls to gain understanding of the expression of glycation-end products (AGEs). N-Carboxymethyl-lysine (CML) and N-carboxyethyl-lysine (CEL) were used as markers of glycoxidation; 4-hydroxynonenal (4-HNE) and malondialdehyde-lysine (MDAL) as markers of lipoxidation; and nitrotyrosine (N-tyr) and neuronal, endothelial and inducible nitric oxide synthase (nNOS, eNos and iNos) as markers of protein nitration and as sources of NO production, respectively. Age receptor (RAGE) and Cu/Zn superoxide dismutase (SOD1) and Mn superoxide dismutase (SOD2) expression levels were also examined. The results showed a significant increase in the expression levels of AGE (p<0.05), CEL (p<0.001), RAGE (p<0.05), HNE modified proteins (p<0.01), nNOS, iNOS and eNOS (p<0.01 and p<0.05, respectively), N-tyr (p<0.05), and SOD1 (p<0.05) and SOD2 (p<0.05). No relationship was observed between PrP genotype, PrP type, PrP burden, and expression levels of oxidative stress markers. The present findings demonstrate oxidative, glycoxidative, lipoxidative and nitrative protein damage, accompanied by increased oxidative responses, in the cerebral cortex in sporadic CJD. These results provide support for the concept that oxidative stress may have important implications in the pathogenesis of prion diseases. PMID- 16310894 TI - [Incidence of cardiovascular events in giant cell arteritis: preliminary results of a prospective double cohort study (GRACG)]. AB - Giant cell arteritis is the most frequent vasculitis. Cardiovascular events such as cerebrovascular accident or ischemic heart disease may occur in patients with giant cell arteritis. However, their real incidence, as well as their relative risk compared to the general population, remains unknown. PURPOSE: To assess in a prospective, double cohort study, the incidence of cardiovascular events in giant cell arteritis patients compared to controls, after controlling for cardiovascular risk factors. PATIENTS AND METHODS: We included on predefined criteria 432 newly diagnosed patients with giant cell arteritis, each assigned to sex- and age-matched controls randomly selected from the general population. Cardiovascular risk factors (high-blood pressure, diabetes, smoking, hypercholesterolemia and preexisting peripheral vascular disease) were collected at inclusion. During the 24-month follow-up, all cardiovascular events were collected. After stratification for cardiovascular risk factors, a log-rank test was performed to compare cases and controls. A parametric survival model was used for multivariate analysis. RESULTS: Cardiovascular events all combined were significantly increased in patients with giant cell arteritis (RR = 2.15 [1.21 3.81], P = 0.009), and were mainly associated with age (P = 0.0001), past history of cardiovascular disease (P = 0.023) but also with giant cell arteritis (P = 0.009). However, each subset of cerebrovascular accident (RR = 2.42 [0.84-7]) or ischemic heart disease (RR = 1.67 [0.72-3.89]) increased but did not significantly. CONCLUSION: Cardiovascular events incidence is increased in patients with giant cell arteritis, and prescription of preventive antiagregant treatment may be discussed. PMID- 16310895 TI - The effect of heparin in peripheral intravenous nutrition via a fine-bore midline: a randomised double-blind controlled trial. AB - BACKGROUND AND AIMS: Peripheral intravenous nutrition (PIVN) delivered via a finebore midline offers a viable alternative to central venous feeding. The major complication is the onset of peripheral vein thrombophlebitis (PVT). Feed additives such as heparin and hydrocortisone have been advocated in its prevention. Concern over the safety of heparin has prevented its widespread use; this study examines its true benefit. METHODS: A randomised, double-blind trial comparing the addition of either, Heparin 1500 units or hydrocortisone 15 mg or a combination of the two to full intravenous nutrition (IVN) (2500 ml, 13 g of nitrogen, 1190 m0sm/k) was performed. All feeds were delivered via a finebore midline inserted via an antecubital fossa vein. Feeding was terminated in the event of complication or cessation of intended nutritional support. RESULTS: One hundred and twenty-three episodes of feeding occurred in 110 patients. The incidence of peripheral vein thrombosis was similar in each group (Heparin 30% (12/41), hydrocortisone 33% (14/42), combination 31% (13/41) chi(2) test P>0.05). There was a significant difference in total catheter survival with the addition of heparin to the feeds, either alone (11 days (SEM 1.79) or in combination with hydrocortisone (11.7 days (SEM 1.39) compared with those receiving only hydrocortisone (6.9 days (SEM 0.73) P=0.002 and 0.030, respectively)). CONCLUSION: When intravenous feeds are delivered in to a peripheral vein via a fine-bore midline, the addition of heparin to the feed extends the total period of feeding attainable. PMID- 16310896 TI - Expression and purification of an influenza hemagglutinin--one step closer to a recombinant protein-based influenza vaccine. AB - Numerous human infections with avian influenza viruses in Asia in recent years have raised the concern that the next influenza pandemic is imminent. The most effective way to combat influenza is through the vaccination of the public. However, a minimum of 3-6 months is needed to develop an influenza vaccine using the traditional egg-based vaccine approach. The influenza hemagglutinin protein (HA), the active ingredient in the current vaccine, can be expressed in insect cells using the baculovirus expression vector system and purified rapidly. An influenza vaccine based on such a recombinant antigen allows a more timely response to a potential influenza pandemic. Here, we report an innovative monitoring assay for recombinant HA (rHA) expression and a rapid purification process. Various biochemical analyses indicate that the purified rHA is properly folded and biologically active. PMID- 16310897 TI - Immunogenicity of a multicomponent DNA vaccine against visceral leishmaniasis in dogs. AB - Vaccination of dogs, the domestic reservoir of Leishmania chagasi, could not only decrease the burden of canine visceral leishmaniasis (VL), but could also indirectly reduce the incidence of human VL. Intramuscular vaccination of foxhounds with a Leishmania multicomponent (10 antigen) DNA vaccine resulted in antigen-induced lymphoproliferative and IFN-gamma (but not IL-4) responses. This response was not augmented by co-administration of canine IL-12 or GM-CSF DNA adjuvants. The multicomponent DNA vaccine also induced a delayed type hypersensitivity (DTH) response to viable L. donovani promastigotes and led to a reduction of parasite burden in an in vitro intracellular infection model, and in the draining lymph node of dogs early after cutaneous challenge. Thus, the multicomponent DNA vaccine was effective in priming dogs for a parasite-specific type 1 cellular immune response, which was able to restrict parasite growth. PMID- 16310898 TI - Topical imiquimod is a potent adjuvant to a weakly-immunogenic protein prototype vaccine. AB - A major challenge in the development of more effective vaccines for cancer and other diseases is the development of potent adjuvants that can strongly, simply and safely enhance vaccine immunogenicity. Adjuvants that preferentially enhance Th1 type of responses are particularly desirable, as these responses are believed to play the major role in immune resistance to cancer. This study describes the ability of topical application of imiquimod to act as a potent, safe and simple vaccine adjuvant in mice. Groups of C57BL/6 mice were immunized subcutaneously with ovalbumin (OVA, 0.1mg/dose) weekly x 4. Imiquimod in a 5% cream formulation was rubbed into the skin over the injection site for 15s to give a dose of approximately 1mg/treatment following each immunization. Control mice were immunized with OVA alone, with irradiated E.G7-OVA cells (that express ovalbumin), with OVA encapsulated in liposomes, or to PBS. Topical imiquimod enhanced anti-OVA antibody responses 100-fold and markedly increased cellular responses compared to mice not given imiquimod. The responses were shifted towards a Th1 phenotype, with marked enhancement of IgG2a, IgG2b, and CD8+ T cell responses and concomitant suppression of IgM and IgG1 responses. More frequent topical applications of imiquimod further enhanced both antibody and cellular responses. There was no detectable local or systemic toxicity associated with treatment. These results indicate that topical imiquimod can safely and strongly enhance both antibody and CD8+ T cell response to OVA immunization, and suggest that it may provide a simple, safe and effective way to enhance the immunogenicity of vaccines in general. PMID- 16310899 TI - Rates of hospitalisation for influenza, respiratory syncytial virus and human metapneumovirus among infants and young children. AB - To inform the development of a national influenza immunisation programme and the potential role of antiviral drugs in young children, we studied 613 children aged 71 months or younger who attended Leicester Childrens' Hospital during winter 2001-2002. During periods of respiratory syncytial virus (RSV), influenza, and human metapneumovirus activity, an estimated 12.2% (95% CI: 11.4-13.1), 7.1% (6.3 7.9), and 2.5% (2.1-2.9), respectively, of all medical cases assessed in the hospital were associated with these infections. The respective rates of hospital assessments for RSV, influenza, and human metapneumovirus (HMPV) were 1042 (95% CI: 967-1021), 394 (348-443), and 223 (189-262) per 100,000 children, and for admissions were 517 (465-574), 144 (117-175), and 126 (101-156) per 100,000. Few children with influenza had a prior risk factor. Children with influenza were admitted a median of 4 days after onset of illness and none was coded at discharge as influenza. We conclude that antivirals have little role in the hospital management of children with influenza. Our data provide health economists with information to evaluate the place of universal immunisation of young children against influenza. Hospitalisation rates decreased markedly with referral age, so vaccine would need to be given in early infancy for maximum benefit. PMID- 16310900 TI - Non PC liposome entrapped promastigote antigens elicit parasite specific CD8+ and CD4+ T-cell immune response and protect hamsters against visceral leishmaniasis. AB - Leishmania donovani promastigote soluble antigens (sLAg) were encapsulated in non phosphatidylcholine (non-PC) liposomes (escheriosomes) prepared from E. coli lipids. The escheriosome-based vaccine was investigated for its potential to elicit a protective immune response against experimental visceral leishmaniasis. The vaccine administration induced strong humoral as well as cell mediated immune responses both in hamsters and BALB/c mice. Immunization of BALB/c mice with escheriosome entrapped sLAg (EL-sLAg) elicited stronger CD8+ cytotoxic T lymphocyte (CTL) response as compared to sLAg entrapped in egg PC/chol liposome (EPC-sLAg) or sLAg administered with incomplete Freund's adjuvant (IFA-sLAg). EL sLAg also induced the release of mixed (Th1 and Th2) types of cytokines in the immunized BALB/c mice. In addition, the delivery of sLAg via escheriosomes enhanced the expression of costimulatory signals (CD80 and CD86) as determined in peritoneal macrophages obtained from BALB/c mice. In another set of experiments, the EL-sLAg immunized hamsters were found to be better protected than those immunized with EPC-sLAg. The prophylaxis coincided with increased lymphocyte proliferation as well as high nitric oxide (NO) production by peritoneal macrophages among EL-sLAg immunized hamsters. Escheriosomes thus seem to have potential in delivering the antigen to cytosol of the antigen presenting cells (APCs) and in the development of liposome-based vaccine against leishmaniasis as well as other intracellular infections. PMID- 16310901 TI - Immunogenicity of a hepatitis B DNA vaccine administered to chronic HBV carriers. AB - Hepatitis B virus (HBV) is the major pathogen of chronic hepatitis and liver disease worldwide. Despite the availability of effective vaccines against hepatitis B for many years, over 370 million people remain persistently infected with HBV. Viral persistence is thought to be related to poor HBV-specific T-cell responses. Based on clinical data, the development of efficient methods capable of inducing strong T-cell responses is an important and primary step toward the development of immunotherapeutics against chronic HBV infection. We designed a phase I clinical trial in chronic HBV carriers to assess safety, tolerability and immunogenicity of a DNA vaccine expressing HBV small (S) and middle (preS2 +S) envelope proteins. After occurrence of lamivudine breakthrough, 10 HBeAg positive patients with chronic hepatitis B were followed longitudinally before, during and after DNA vaccine therapy. Immunizations were well tolerated and adverse physical events were mild and considered unrelated to the vaccine. Proliferative responses to hepatitis B surface antigen (HBsAg) were detected in two patients after DNA injections. Following three injections of vaccine, interferon (IFN)-gamma producing T-cells specific for the preS2 or the S antigen were detectable in 50 and 100% of the patients, respectively. Each patient recognized at least one peptide within the envelope domain encoded by the vaccine. Anti-preS2 antibodies and seroconversion to anti-HBe were detected in two patients. This study shows evidences for the safety and immunological efficacy of HBV-DNA vaccination and demonstrates that DNA vaccination can restore or activate T-cell responses in chronic HBV carriers. PMID- 16310902 TI - Hepatitis B revaccination in healthy non-responder Chinese children: five-year follow-up of immune response and immunologic memory. AB - To assess persistence of anti-HBs and immunologic memory of non-responders after revaccination, 40 healthy non-responder children were given a three-dose recombinant hepatitis B vaccine revaccination randomly by intramuscular (10 microg per dose) or intradermal (2 microg per dose) route and followed up to five years. All 17 intramuscular and 22 of 23 intradermal children developed a seroprotective antibody response (anti-HBs>or=10 mIU/mL) after revaccination. Children of intramuscular group had significantly higher seroprotection rates and anti-HBs geometric mean titers than the intradermal group. At year 5, 50% of children in intramuscular group, but only 18.2% of intradermal group still maintained seroprotection (P=0.075). By the end of follow-up, a booster dose (5 microg) was given to those who had lost seroprotection. All the eight intramuscular children developed an anamnestic response with increase of anti-HBs level by 215 times, but two of the 18 intradermal children failed to produce seroprotective level. Three-routine-dose intramuscular revaccination was significantly more effective than low-dose intradermal revaccination with the same number of injections. No child seroconverted to HBsAg, and 11 had transient infections indicated by seroconversion to anti-HBc. These results demonstrated that non-responders could benefit from three doses intramuscular revaccination not only in high proportion of anti-HBs conversion but also in long-term persistence of seroprotection, and more importantly in preservation of the immunologic memory years after loss of protective anti-HBs. PMID- 16310903 TI - Purification of phocid herpesvirus type 1 glycoproteins B and D and pilot studies of immunogenicity in mice. AB - In the process of developing a subunit vaccine against phocid herpesvirus type 1, we have cloned and expressed the glycoproteins B and D (gB and gD) of phicid herpesvirus type 1, using an eukaryotic baculovirus expression system. To establish the proof of concept, candidate iscom vaccines based on these affinity purified proteins either alone or in combination, were tested for their immunogenicity in BALB/C mice. Mice immunised with a combination of gB and gD developed higher antibody and proliferative T cell responses against PhHV-1 than those immunised with gB or gD alone. The corresponding antibody and T cell proliferative responses were higher against PhHV-1 than against FHV. These data favour further testing of these candidate vaccines based on gB and gD in the FHV cat model. PMID- 16310904 TI - Interesting case: an unusual penetrating injury--the forked tongue. PMID- 16310905 TI - Orbital abscess after extraction of a maxillary wisdom tooth. AB - A 21-year-old patient with orbital abscess and vision loss left after wisdom tooth extraction is presented. PMID- 16310906 TI - Results of the centralisation of adult emergency oral and maxillofacial surgical services at the University Hospital, Birmingham. AB - In 2001, the oral and maxillofacial surgical (OMFS) services for adults in Birmingham were centralised at the Selly Oak site of the University Hospital and the OMFS department was given access to the emergency operating theatre every day. We examined the effects of this on the emergency workload during the 6 months after centralisation and compared it with the emergency workload at the University Hospital during the 6 months before centralisation. The number of adult patients who attended the University Hospital as emergencies increased from 242 in the 6 months before centralisation to 545 in the subsequent 6 months (an increase of 127%). Of the latter 164 (30%) were admitted and operated on, 138 (84%) within 24h of admission. Despite the large increase in the number of patients, 102 (74%) were operated on during normal working hours. After centralisation, all operations were done on multidisciplinary emergency lists compared with only 55% before centralisation. Centralisation did not reduce the access of patients to the emergency service and conformed with the guidelines of the National Confidential Enquiry into Perioperative Deaths (NCEPOD). PMID- 16310907 TI - The aggressive form of cherubism: report of two cases in unrelated families. AB - Cherubism is a benign lesion that causes painless symmetrical enlargement of the jaws, usually with a familial tendency. We describe in two Chinese families two cases of the aggressive form of cherubism with extensive swelling on both sides of the mandible, typical microscopic findings, and apparent familial history. PMID- 16310908 TI - Exposure of implants using a modified multiple-flap transposition vestibuloplasty. AB - AIM: To introduce a minimally invasive operation to improve the condition of the soft tissues around the implants in an atrophied mandible, at the same time, as uncovering the implants. PATIENTS AND METHOD: A multiple-flap transposition vestibuloplasty was done in 11 patients after the insertion of four implants in the interforaminal region of an atrophied mandible. Improvement in soft tissues and successful exposure of implants and attached gingiva were evaluated during a follow-up period of 55 months. All the patients were operated on local anaesthesia as outpatients. RESULTS: Adequate exposure of implants and an area of attached gingiva 4-9 mm wide were attained. There was no bleeding on probing or local infection. CONCLUSION: The transposition multiple-flap vestibuloplasty is a simple and minimally invasive method of improving the condition of soft tissue after insertion of implants. It does not limit the patients' routine activities and avoids staged operations. PMID- 16310909 TI - Interesting case: The importance of the history in patients with haemangioma of the head and neck. PMID- 16310910 TI - Scapholunate dislocation: a rare occupational hazard. AB - Dentists and their assistants are at risk of physical injuries during dental operations. The most common injuries are musculoskeletal. The need to work in a fixed working position using continuous repetitive movements can predispose dentists to wrist ache, lower backache, and neckache. We encountered a rare case of scapholunate dislocation with ligamentous injury, an unusual complication of extraction of teeth. PMID- 16310911 TI - Risk of plantar ulceration in diabetic patients with single-leg amputation. AB - BACKGROUND: There is a disconcerting rate of bilateral limb loss in patients with diabetes. Therefore, this study aimed to explore plantar loading of the surviving foot following unilateral trans-tibial amputation within a wider context of daily walking activity to investigate the precise risk to the surviving limb. METHODS: Twenty-one subjects with diabetic neuropathy and trans-tibial amputation were matched for weight; height; age and gender with 21 control subjects with diabetic neuropathy without history of plantar ulceration. Gait parameters, in-shoe plantar pressure distribution and daily walking (using the step activity monitor) were recorded. Student's t-tests were used to compare groups (alpha-level: 0.05). FINDINGS: The trans-tibial amputations group walked almost 30% slower compared to controls (P < 0.01), with reduced cadence (P < 0.01), and shorter strides (P < 0.01). Despite walking slower, the surviving foot showed higher mean peak plantar pressures in the trans-tibial amputations group over the heel (P < 0.001) however there was no significant difference over the I-II and lateral III-IV-V metatarso phalangeal regions. Pressure time integral was higher over the heel (P < 0.00), I II (P < 0.01) and III-IV-V metatarso-phalangeal (P < 0.05) in the trans-tibial amputations group. The amputee group walked less steps per day (P < 0.01). INTERPRETATION: Adaptations in gait and level of walking activity affect plantar pressure distribution and ultimately the risk of ulceration to the surviving foot. Therefore rehabilitation measures should consider implications for plantar loading and the potential risk of ulceration to the surviving foot. PMID- 16310912 TI - Proceedings of the 5th meeting of the International Shoulder Group, August 2004, Estoril, Portugal. PMID- 16310913 TI - Consequences for Protaphorura armata (Collembola: Onychiuridae) following exposure to genetically modified Bacillus thuringiensis (Bt) maize and non-Bt maize. AB - Studies on the effect of genetically modified Bacillus thuringiensis (Bt) crops on true soil dwelling non-target arthropods are scarce. The objective of this study was to assess the influence of a 4-week exposure to two Bt maize varieties (Cry1Ab) Cascade and MEB307 on the collembolan Protaphorura armata. For comparison three non-Bt maize varieties, Rivaldo (isogenic to Cascade), Monumental (isogenic to MEB307) and DK242, and two control diets based on baker's yeast (uncontaminated and contaminated with Bt toxin Cry1Ab) were also tested. Due to a lower C:N ratio, individuals reared on yeast performed significantly better in all of the measured endpoints than those reared on maize. P. armata performed equally well when reared on two Bt and three non-Bt maize varieties. Although there were no negative effects of Bt maize in this experiment, we recommend future studies on Bt crops to focus on species interactions in long term, multi-species experiments. PMID- 16310914 TI - Metal stoichiometry in predicting Cd and Cu toxicity to a freshwater green alga Chlamydomonas reinhardtii. AB - In this study, we quantified the accumulation and toxicity of cadmium and copper in a freshwater green alga, Chlamydomonas reinhardtii, under different phosphate conditions. The accumulated Cd and Cu concentrations increased significantly with increasing ambient P concentrations and free metal ion concentrations. The metal:P ratio remained independent of the ambient P concentration. For the three pulse-amplitude-modulated parameters, the median inhibition concentrations were 1.5-1.6x and 2.0x higher, but the medium inhibition cellular quota was 2.2x and 1.2x lower for cells maintained at 0.1 microM P than for cells maintained at 10 microM P for Cd and Cu, respectively. Furthermore, the difference in metal toxicity decreased (for Cd) or disappeared (for Cu) when the toxicity was expressed by the metal:P ratio in the cells, indicating that the stoichiometry of metals and P can be better used to predict the toxicity of metals. It is necessary to consider the stoichiometry of metals in predicting metal toxicity in phytoplankton. PMID- 16310915 TI - Environmental contamination of mercury from Hg-mining areas in Wuchuan, northeastern Guizhou, China. AB - Total Hg and methyl-Hg were evaluated in mine wastes, soils, water, and vegetations from the Wuchuan Hg-mining areas, Guizhou, China. Mine wastes contain high total Hg concentrations, ranging from 79 to 710 microg g(-1), and methyl-Hg from 0.32 to 3.9 ng g(-1). Total Hg in soil samples range from 0.33 to 320 microg g(-1) and methyl-Hg from 0.69 to 20 ng g(-1). Vegetations present a high average total Hg concentration of 260 ng g(-1), which greatly exceeds the maximum Hg concentration of 20 ng g(-1) recommended by the Chinese National Standard Agency for food sources. The rice samples contain elevated methyl-Hg concentrations, ranging from 4.2 to 18 ng g(-1). Stream water collected from Hg-mining areas is also contaminated, containing Hg as high as 360 ng l(-1), and methyl-Hg reaches up to 5.7 ng l(-1). Data indicate heavy Hg-contaminations and significant conversion of methyl-Hg in the study areas. PMID- 16310916 TI - "Does it mean I'm gonna die?": on meaning assessment in the delivery of diagnostic news. AB - This paper investigates how, in the delivery of diagnostic news, participants to the delivery may engage in meaning assessment or interpreting the news. It draws on data from 24 conversations in developmental disabilities clinics, internal medicine clinics and HIV counselling and testing clinics in the USA. The analysis shows that participants initiate meaning assessment sequences whereby one participant proposes what the news means and the other aligns or disaligns with the proposal. When meaning assessment occurs, the preferred way for this to happen is that the clinician initiates and proposes an interpretation. Following the interpretive proposal, a patient or family member aligns or disaligns with the interpretation, with alignment being sought over disalignment. Further practices of meaning assessment are "affirming the positive" and "disconfirming the negative," which work to provide relatively benign interpretations of news. Analysis of a collection of meaning assessment sequences in clinical settings is brought to bear on a single case in which an internist tells a patient that he has stomach cancer. After delivering the diagnosis, this doctor neither affirms the positive nor disconfirms the negative, and the patient ends up asking, "Does it mean I'm gonna die?" At this point, the interview gets disrupted as the patient withdraws. Asking what the news means is a structurally dispreferred way of handling problems of meaning, and as such this patient's exhibition of difficulty is an outcome of orderly social practices. A clinician's withholding of auspicious meaning assessment may undermine the relationship with patients and/or family members and disrupt the encounter. PMID- 16310917 TI - Chronic nicotine interactions with clozapine and risperidone and attentional function in rats. AB - Although antipsychotic drugs are therapeutically effective in attenuating the hallmark symptoms of schizophrenia, these improvements do not return most patients to normative standards of cognitive function. Thus, complementary drug treatment may be needed to treat the attentional deficits of schizophrenia as well as to counteract the potential attentional impairments caused by some antipsychotic drugs. Nicotine, a drug commonly self-administered by a great majority of individuals with schizophrenia, has been shown to significantly improve cognitive function in some studies. The current study was conducted to determine the interactive effects of the atypical antipsychotic drugs clozapine and risperidone with chronic nicotine administration on attentional performance. Adult female Sprague-Dawley rats (N=35) were trained to perform an attentional task using an operant visual signal detection task. After training, rats were infused with a dose of 5 mg/kg/day (s.c.) nicotine base (n=18) or saline (n=17) for 28 consecutive days via osmotic pump. In Exp. 1, while being administered chronic nicotine or saline, rats were given acute doses of clozapine (0, 0.625, 1.25 and 2.5 mg/kg, s.c.) and were tested for attentional function. In Exp. 2, while on chronic nicotine or saline, other rats were challenged with acute doses of risperidone (0, 0.025, 0.05 and 0.1 mg/kg, s.c.) and were tested for attentional function. Results showed that acute administration of clozapine caused a significant dose-dependent impairment in choice accuracy (percent hit) in animals treated with chronic saline. Chronic nicotine treatment itself lowered accuracy, but attenuated further declines with acute clozapine treatment. Acute administration of risperidone at high dose significantly reduced performance (percent correct rejection) in chronically saline-treated rats, but in a similar fashion as in Exp. 1, chronic nicotine lowered accuracy but attenuated further impairment with acute risperidone. In summary, atypical antipsychotic drugs clozapine and risperidone significantly impaired choice accuracy in the visual signal detection task. Clozapine was more detrimental than risperidone but the adverse effects of both clozapine and risperidone on attentional performance were masked in rats chronically treated with nicotine. PMID- 16310918 TI - Opposite effects of depression and antidepressants on processing speed and error rate. AB - Depressions increase reaction times and antidepressant treatment decreases them. Is it possible to get information, how these two conditions affect the stimulus response pathway? Therefore, auditory and visual, left-hand and right-hand, simple and choice reaction times were measured in 38 patients with depression and 30 healthy control subjects. From these data, the minimal reaction times and the error rates were calculated. In the first study, the reaction times and error rates of patients with depression were correlated with the antidepressant doses. In the second study, the differences between medicated, depressive patients and healthy control subjects were investigated. The result of the first study was that antidepressant doses correlated negatively with reaction times but positively with error rates. In the second study, the patients with depression showed increased reaction times but reduced error rates compared to healthy control subjects. The auditory tasks were more affected than the visual tasks and the minimal reaction times more than the median reaction times. Therefore, we concluded that depression and antidepressant treatment had opposite effects on reaction times and error rates. The findings in minimal reaction times and auditory tasks suggest a special involvement of early cortical auditory processing in depression and antidepressant treatment. PMID- 16310919 TI - Cataleptic activity of the denatured venom of the social wasp Agelaia vicina (Hymenoptera, Vespidae) in Rattus norvegicus (Rodentia, Muridae). AB - Catalepsy is a state of immobility, commonly experienced by patients with chronic use of many antiparkisonism and neuroleptic drugs. Recently, catatonia has been considered as an evolutionary-based fear response. The cataleptogenic effects of the low molecular weight compounds from the venom of the social wasp A. vicina (AdAv) were reported in rodents. Intracerebroventricular injections of AdAv in highest dose reduced the locomotor activity of Wistar rats in the open field and induced ataxia and catalepsy within 10 min. This effect could be observed up to 30 min after injections. Lower doses of denatured venom injected in brain ventricles also reduced the locomotor activity of the rats but did not induce catalepsy. The cataleptic effects of the intracerebrally administered AdAv were antagonized by the peripheral (intraperitoneal) pretreatment with theophylline and ketamine. Moreover, the central effects of the AdAv were compared to those elicited by the neuroleptic drug haloperidol (intraperitoneally administered), whose cataleptic effects were also antagonized by theophylline and ketamine. However, the association of haloperidol and denatured venom was reverted by theophylline but not by ketamine. These findings suggest that A. vicina venom may affect neural substrates involved with catalepsy in the central nervous system. PMID- 16310920 TI - Effect of sodium hypochlorite on primary dentin--a scanning electron microscopy (SEM) evaluation. AB - OBJECTIVE: The aim of this study was to evaluate the alterations of etched deciduous dentin when submitted to different time and concentrations of sodium hypochlorite (NaOCl), using scanning electron microscopy (SEM). MATERIAL AND METHODS: Forty deciduous anterior teeth were selected, cleaned and ground until expose a flat dentin area on the buccal surface. The specimens were randomly distributed into eight groups (n=5), according to dentin surface treatment (35% phosphoric acid etching for 7s--AE and/or NaOCl application), NaOCl solution concentration (5% or 10%), and time of application (0, 30, 60, and 120 s), as follows: G1: control (without AE and NaOCl); G2: only AE; G3, G4, and G5: AE+5% NaOCl for 30, 60, and 120 s, respectively; G6, G7, and G8: AE+10% NaOCl for 30, 60, and 120 s, respectively. All specimens were prepared for SEM analysis and the photomicrographs (three for each specimen) were classified according to a score as follow: 0: presence of smear layer (SL); 1: absence of SL+non-altered collagen fibrils; 2: absence of SL+collagen fibrils slightly altered; 3: absence of SL+collagen fibrils severely altered; and 4: absence of SL and absence of collagen fibrils. Data were submitted to Kruskal-Wallis and Mann-Whitney tests (p<0.05). RESULTS: All groups treated with NaOCl solution were significant different from G1 and G2, and showed alterations on the collagen fibrils network. Collagen complete removal was only observed when a 5% NaOCl solution was applied for 120 s and 10% NaOCl solution for 30, 60, and 120 s. CONCLUSIONS: The NaOCl action produced significant changes in the etched deciduous dentin. The higher NaOCl concentration, the lower the time required to completely removing the collagen fibrils network in deciduous dentin. PMID- 16310921 TI - Characterization of the 5'- and 3'-ends of mRNAs of ndhH, ndhA and ndhI genes of the plastid ndhH-D operon. AB - The monocistronic mRNAs of ndhH and ndhA genes of the plastid ndhH-D operon accumulate during senescence. In a step forward to discover the mechanisms involved in the production of mature transcripts of the ndhH-D operon, we determined the 5 - and 3 -ends of low-molecular-weight ndhH, ndhA and ndhI transcripts of barley. The 3'-end of the only ndhH mRNA detected (1622 b) extends 421 b into exon1 of ndhA. Several monocistronic ndhA mRNAs were identified and most of them extend their 3'-ends through the adjacent intergenic 101 b region to around 105 b into ndhI, the neighboring gene in the operon. Unspliced ndhA transcripts (around 2.4 kb and intron-containing) were also identified with different 3'-ends extending almost 200 b into ndhI. In contrast, all ndhA transcripts showed the same 5'-end 65 b upstream of the ndhH stop codon. 5'-ends of all ndhA and ndhH transcripts seem produced by nuclease cleavages between the last AA of the consensus sequence AAUGAA present 66 and 16 b upstream of the respective start codon. Secondary structure predictions suggest that 3'-end extensions provide 3'-UTR stabilizing elements to ndhH-D transcripts. Therefore, the processing of the 7.8 kb primary transcript frequently involves intra-genic cleavages sacrificing downstream gene messages and decreasing the efficiency of the conversion to mature ndh mRNAs. Significantly, the maturation of ndhE mRNA would not sacrifice the downstream psaC message, contributing to higher levels of psaC mRNA than of ndh mRNAs. PMID- 16310922 TI - Nitric oxide inhibits HIF-1alpha protein accumulation under hypoxic conditions: implication of 2-oxoglutarate and iron. AB - Cells exposed to low oxygen conditions respond by initiating defense mechanisms, including the stabilization of hypoxia-inducible factor (HIF) 1alpha, a transcription factor that upregulates genes such as those involved in angiogenesis and glycolysis, which also plays a pivotal role in the regulation of cellular utilization of oxygen and is an essential regulator of angiogenesis in solid tumor and ischemic disorders. Nitric oxide and other inhibitors of mitochondrial respiration prevent the stabilization of HIF-1alpha during hypoxia. In the present study we found that nitric oxide inhibits HIF-1alpha accumulation under low oxygen (1%) conditions. The effect is supported by an increase in 3 nitrotyrosine and is more likely caused by the formation of peroxynitrite in the cells, which leads to the damage of mitochondria and their respiratory chain followed by the increase in 2-oxoglutarate (2-OG) and iron (the components needed to activate HIF-1alpha proline hydroxylases) concentrations in cell cytosol. The inhibiting effect of NO on HIF-1alpha accumulation was not observed in the cells lacking mitochondria. On the other hand the depletion of intracellular glutathione (GSH) was observed upon cell treatment with nitric oxide donors under hypoxic conditions. Treatment of those cells with N-acetyl cysteine (NAC) increased the amount of intracellular GSH and attenuated the NO effect and abolished the damage of mitochondria as well as 2-OG/iron release. PMID- 16310923 TI - Editing of the grapevine mitochondrial cytochrome b mRNA and molecular modeling of the protein. AB - Cytochrome b (COB), the central catalytic subunit of ubiquinol cytochrome c reductase, is a component of the transmembrane electron transfer chain that generates proton motive force. Some plant COB mRNAs are processed by RNA editing, which changes the gene coding sequence. This report presents the sequences of the grapevine (Vitis vinifera L.) mitochondrial gene for apocytochrome b (cob), the edited mRNA and the deduced protein. Grapevine COB is 393 amino acids long and is 98% identical to homologs in rapeseed, Arabidopsis thaliana and Oenothera sp. Twenty-one C-U editing sites were identified in the grapevine cob mRNA, resulting in 20 amino acid changes. These changes increase the overall hydrophobicity of the protein and result in a more conserved protein. Molecular modeling of grapevine COB shows that residues changed by RNA editing fit the secondary structure characteristic of an integral membrane protein. This is the first complete mitochondrial gene reported for grapevine. Novel RNA editing sites were identified in grapevine cob, which have not been previously reported for other plants. PMID- 16310924 TI - BXL628, a novel vitamin D3 analog arrests prostate growth in patients with benign prostatic hyperplasia: a randomized clinical trial. AB - OBJECTIVE: To evaluate the effect of BXL628, a vitamin D3 analog, on prostate volume in patients with benign prostatic hyperplasia (BPH). METHODS: We conducted a phase II, double blind, randomized, placebo controlled, clinical study. Patients eligible were aged>or=50 years, had a diagnosis of BPH and a prostate volume>or=40 ml. Eligible patients were randomized and given either BXL628 150 mcg daily or placebo for 12 weeks. All randomized patients underwent at baseline and at the end of study pelvic MRI to measure prostatic volume, uroflowmetry (Qmax), American Urological Association Symptom Index (AUASI), serum PSA, testosterone, dihydrotestosterone and luteizing hormone. RESULTS: A total of 119 patients were randomized: 57 patients to BXL628 and 62 to placebo. The percentage change of prostate volume at 12 week was -2.90 in the BXL628 group vs. +4.32 in the placebo group (p-value<0.0001). The estimated difference between treatments (BXL628 minus placebo) was -7.22% (95% confidence limit -9.27 to -5.18). Considering Qmax, mean change vs. baseline was -0.30 in BXL628 vs. +1.50 in the placebo group: this finding was not statistically significant. The mean change of the AUASI total score at final visit vs. baseline was -1.77 in the BXL628 group vs. -3.45 in the placebo group (p=not significant). CONCLUSION: BXL628 was able to arrest prostate growth within 12 weeks in men aged>or=50 years with prostatic volume>or=40 ml. Its unprecedented mechanism of action may offer a new opportunity for the treatment of BPH. PMID- 16310925 TI - Lack of prophylactic effect of incadronate on skeletal lesions associated with implants of prostate cancer. AB - OBJECTIVES: To evaluate prophylactic effects of bisphosphonate (BP) on skeletal lesions induced by prostate cancer (CaP). METHODS: Incadronate, a third generation BP, was administered weekly for 4 weeks, with or without 4-week pre administration, to rats whose calvaria had been inoculated with syngeneic CaP tissue. RESULTS: The transplanted CaP grew up and caused bone resorption with osteoblastic changes regardless of incadronate treatment. Although decrease in bone resorption accompanied by reduced number of osteoclasts was shown by incadronate administration, pre-administration had no additional inhibitory effect on bone destruction and Ki-67 labeling indices of CaP cells were not altered. CONCLUSIONS: These results indicate that application of BP alone is not sufficient to prevent skeletal lesions due to CaP in patients with high risk of bone metastases although it is useful in inhibition of cancer-induced bone resorption with osteoblastic changes. PMID- 16310926 TI - New perspectives of penile enhancement surgery: tissue engineering with biodegradable scaffolds. AB - OBJECTIVE: To evaluate in a multicenter, prospective study preliminary aesthetic and functional results of autologous ex-vivo tissue engineering for penile girth enhancement. METHODS: From July 1999 to January 2004, 204 men of mean age 26.77 (range 19-54 years) underwent this procedure. Indications for penile girth enhancement were penile dysmorphic disorder and previous failed surgery for penile girth enhancement. Fibroblast cells harvested from 1 cc of biopsied scrotal dermal tissue were expanded in culture until the total cell number of at least 2x10(7) was reached. Suspended cells in culture medium were then seeded on pretreated tube-shaped PLGA scaffolds and incubated for 24 hours. After penile degloving, scaffolds were shape adjusted and transplanted between dartos and Buck's fascia when the skin was compliant or under the neurovascular bundle when the skin was not compliant. RESULTS: A total of 84 randomly selected patients were followed 1 to 5 years postoperatively (median 24 months). The gain in girth ranged from 1.9 to 4.1cm (mean 3.15 cm). Postoperative complications occurred as infection in three, penile skin pressure necrosis in two and seroma formation in five patients and were all treated conservatively. Surgical intervention was appraised by patients on a scale from 1 to 5 as follows: the best mark (5) was given by 44.05%, very good (4) by 36.90%, good (3) by 19.05% and only one patient gave the mark 2 judging general penile appearance as dissatisfactory; mean score was 4.25. CONCLUSION: Autologous tissue engineering by using biodegradable scaffolds as a carrier is a new and safe therapeutic approach for penile girth enhancement. The outcome of this study points out the necessity for its expanded clinical applicability in the future. PMID- 16310927 TI - Transcapsular adenomectomy(Millin): a comparative study, extraperitoneal laparoscopy versus open surgery. AB - OBJECTIVE: To demonstrate the safety of the Millin extraperitoneal laparoscopic adenomectomy by comparing the laparoscopic and open approaches. METHODS: From January 2003 to April 2005, patients, with indication of prostatic adenomectomy, were offered the chance to choose between 2 types of procedure: Millin adenomectomy with open or extraperitoneal laparoscopic approach. Forty patients were included in this prospective non-randomised study. The patients were divided into 2 Groups. Group A (20 patients) underwent extraperitoneal laparoscopic adenomectomy, while Group B (20 patients) was treated by open surgery. Pre operative, peri-operative and post-operative parameters were evaluated. Group A was subdivided in two sub-groups (first ten and last ten) and peri-operative parameters were considered in order to determine the learning curve. RESULTS: In terms of pre-operative parameters considered, the 2 study groups are comparable (p>0.3). As far as peri and post-operative parameters are concerned: mean blood loss for Group A was 411.6+/-419 ml, for Group B 687.5+/-298.6 ml (p=0.004). For all the other parameters no significant statistical differences were recorded (p>0.4). Mean operative time was 107.2+/-34.9 min in Group A, and 95.5+/-22.5 min in Group B. Mean adenoma weight in Group A was: 69,5+/-21.5 g, in Group B: 88.1+/ 43.8 g. Mean haemoglobin levels in Group A was: 11.2+/-1.8 g/dl, Group B: 11.6+/ 1.2 (10-13.4) g/dl. Mean Analgesic consuming (Tramadol) during the post-operative stay was 385+/-36 mg in Group A, versus 430+/-108 mg in Group B. Mean catheterization time was 6.3+/-3.7 days in Group A, 5.6+/-1.1 days in Group B. The mean hospital stay was 7.8+/-4.1 days in Group A, and 7+/-1.6 days in Group B. One patient (5%) from Group A was re-operated for bleeding and clot retention, whilst in Group B patients did not present complications which required any further intervention. As far as peri-operative and post-operative parameters of the two sub-Groups A (first ten patients and last ten patients) are concerned, the statistical evaluation shows a significant difference only on operative time (p=0.01). The p-value for the other parameters was not significant (p>0.1). CONCLUSIONS: The extraperitoneal laparoscopic adenomectomy is a safe technique presenting results comparable to open surgery with the advantage of significantly lower peri-operative blood loss. PMID- 16310928 TI - Urinary hyaluronan as a marker for the presence of residual transitional cell carcinoma of the urinary bladder. AB - OBJECTIVE: The purpose of this report is to evaluate the value of urinary hyaluronan (HA) as a maker of residual transitional cell carcinoma (TCC). PATIENTS AND METHODS: Urine samples were collected from 83 patients hospitalized for transurethral resection (TUR). Patient ages ranged from 36 to 86 years. Samples were taken both before and after surgery. HA analysis was performed using an "ELISA-like" fluorometric assay. RESULTS: Patients were divided into two groups: a control group whose previous diagnosis was negative for tumors (n=22) and another with positive diagnosis for tumors (n=61) which was further sub divided into with and without residual tumor. After the second procedure 47 individuals did not display residual tumor, whereas 14 (23%) did. The average HA in the control group was 8.3 microg/L pre- and 7.1 post-operatively, hence, no change occurred (p=0.471). In the group with TCC patients, the HA dropped from 885.5 microg/L to 215.3 microg/L with residual tumors and from 234.3 microg/L to 11.2 microg/L for those without residual tumor. Using a cut-off value of 20 microg/L, the sensitivity to detect residual tumor is 92.9% and specificity is 83%. CONCLUSION: HA in addition to being one of the best markers for the initial evaluation of bladder carcinoma can be used to determine the presence of a residual tumor. This is associated with poor prognosis. PMID- 16310929 TI - Interferon alpha 2b as medical selection for nephrectomy in patients with synchronous metastatic renal cell carcinoma: a consecutive study. AB - OBJECTIVE: Up to 25% of the patients with synchronous metastatic renal cell carcinoma (mRCC) treated with nephrectomy and interferon alpha-2b (IFN-alpha) will progress rapidly at metastatic sites and undergo needless surgery for an asymptomatic primary. We reversed the timing of surgery and immunotherapy and evaluated the role of initial IFN-alpha as selection for nephrectomy. PATIENTS AND METHODS: Sixteen patients with mRCC and the primary in-situ received initial IFN-alpha for 8 weeks (2 weeks 5x3x10(6)IU/wk; 2 weeks 5x6x10(6)IU/wk; 2 weeks 5x9x10(6)IU/wk and 2 weeks 3x9x10(6)IU/wk). Patients with either partial remission (PR) or stable disease (SD) underwent nephrectomy followed by IFN-alpha maintenance at 3x9x10(6)IU/wk. Patients were evaluated with regard to age, sex, metastatic sites, morbidity, response, nephrectomy rate, time to progression and survival. RESULTS: Thirteen patients received 2 months of preoperative IFN-alpha; 3 stopped during the 2 months period due to progressive disease (PD). Eight patients developed either a PR (n=3) or SD (n=5) at metastatic sites and underwent nephrectomy. Survival at 1 year is 50% (4/8 patients). Median progression-free survival was 6 months (3-17 months). Two of the 3 patients with PR developed a CR after 2 months maintenance following surgery. Eight patients with PD did not undergo surgery and had a median survival of 4 months (range 1-8 months). CONCLUSIONS: Absence of progression at metastatic sites following IFN alpha with the primary tumor in place may be used as selection for nephrectomy in patients with an intermediate prognosis. Currently, a randomized study is underway to assess the role of initial versus delayed nephrectomy in combination with IFN-alpha with regard to morbidity and survival. PMID- 16310930 TI - Re: Chapple CR, Martinez-Garcia R, Selvaggi L, Toozs-Hobson P, Warnack W, Drogendijk T, Wright DM, Bolodeoku J. A comparison of the efficacy and tolerability of solifenacin succinate and extended release tolterodine at treating overactive bladder syndrome: results of the STAR trial. Eur Urol 2005;48:464-70. PMID- 16310931 TI - p53 protein transduction therapy: successful targeting and inhibition of the growth of the bladder cancer cells. AB - INTRODUCTION: Virus-mediated gene therapy for bladder cancer has some problems, such as efficiency of gene delivery and safety issues. We have reported that poly arginine peptide (11R) has the ability to increase protein transduction in cells. Here, we show that p53 protein transduction using 11R is useful for targeting to bladder tumors and suppressing the growth of bladder cancer cells. MATERIALS AND METHODS: An 11R-fused p53 protein (11R-p53) was transduced in bladder cancer cell lines (J82 and T24) to evaluate the anti-tumor effect. Cell viability was assessed by performing the 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio] 1,3-benzene disulfonate (WST) assay. To investigate whether 11R-p53 enhanced the effect on anti-cancer drug-dependent apoptosis of bladder cancer cells, the cell lines were cotreated with 11R-p53 and cis-diaminedichloroplatinum (CDDP). Apoptotic cells were identified using Hoechst staining. To investigate the efficiency of protein transduction mediated by 11R in bladder tumors in vivo, SCID mice were transplanted with J82 cells in the bladder and 11R-GFP was transurethrally transduced into the bladder. The transduction of 11R-GFP in the tumor was examined by confocal microscopy. RESULTS: 11R-p53 inhibited the growth of both J82 and T24 cells in a dose-dependent manner. The transduction of 11R-p53 enhanced CDDP-dependent induction of apoptosis. Transurethral application of 11R GFP resulted in transduction of GFP in bladder tumors but not in the normal bladder epithelium or subepithelial tissues. CONCLUSION: The present results suggest that p53 protein transduction therapy may be a promising method for the treatment of bladder cancer. PMID- 16310932 TI - Peri-operative complications and pain after the suburethral sling procedure for urinary stress incontinence: a French prospective randomised multicentre study comparing the retropubic and transobturator routes. AB - OBJECTIVE: To compare peri-operative complications, pain, and the immediate functional results of the sub-urethral sling procedure for urinary stress incontinence by the retropublic and transobturator routes, using a non-elastic polypropylene sub-urethral sling. PATIENTS AND METHODS: This prospective, multicentre study involved 88 women undergoing the sub-urethral sling procedure for stress urinary incontinence (SUI). The retropubic route (RPR) and the transobturator route (TOR) were used in respectively 42 and 46 cases. The characteristics of the women in the RPR and TOR groups were as follows: mean age (+/-standard deviation) 56.8+/-12 years and 53.4+/-10 years, respectively; mean BMI: 25+/-4 and 26+/-4; mean parity: 2.1+/-0.9 and 2+/-1 children; post menopausal status: 66.7% and 58.7%; prior surgery for SUI: 7.1% and 6.5%; and prior hysterectomy: 21.4% and 26.1%. None of these characteristics differed significantly between the groups. Likewise, pre-operative urinary functional status (SUI stage, and pollakiuria, nocturia and urgency rates) was similar in the two groups. RESULTS: Mean hospital stay and overall morbidity rate were not significantly different between the RPR and TOR groups. Mean operating time was longer in the RPR group. Bladder injury was significantly more frequent in the RPR group and vaginal injury was significantly more frequent in the TOR group. Pain scores were significantly lower in the TOR group. The objective functional results at one month did not differ between the groups. Quality of life, evaluated with questionnaires and numerical rating scales, was similarly improved in the two groups. DISCUSSION: The suburethral sling procedure was less painful by the TOR route than by the RPR route. Bladder injury, haematomas and abscesses were only observed in the RPR group, while vaginal injury only occurred in the TOR group. The immediate functional results of the two approaches were similar. PMID- 16310933 TI - Relationship of NKX3.1 and MYC gene copy number ratio and DNA hypomethylation to prostate carcinoma stage. AB - OBJECTIVE: High stage prostate cancers have been reported to frequently harbor chromosome 8 alterations and hypomethylation of LINE-1 retrotransposons. The potential of these parameters for molecular staging of prostate carcinoma was investigated. METHODS: High molecular weight DNA was extracted from 63 carcinoma tissues (22 pT2, 38 pT3, 3 pT4). Chromosome 8 alterations were followed by determining the ratio of NKX3.1 (at 8p21) to MYC (at 8q24) gene copy numbers (NKX3.1:MYC ratio) using a new real-time PCR technique. LINE-1 hypomethylation was quantified by Southern blot analysis. RESULTS: In 42 carcinomas NKX3.1 copy numbers were altered, with decreases in 32 cases. Copy numbers of MYC were increased in 38 cases and diminished in four. The NKX3.1:MYC ratio was altered in 45 specimens, with a decrease in all but two. NKX3.1 loss was associated with tumor stage (p<0.03) and MYC gain with Gleason score (p<0.03). The NKX3.1:MYC ratio was highly significantly associated with tumor stage (p<0.002), displaying 66% sensitivity and 87% specificity. LINE-1 hypomethylation was related (p<0.004) to tumor stage, but exhibited lower sensitivity (59%) and specificity (77%). CONCLUSION: A straightforward PCR technique detecting chromosome 8 alterations might be useful to predict which prostate cancers are organ-confined while determination of hypomethylation appears to be somewhat less well suited. PMID- 16310934 TI - Prostate cancer: fine tuning our ability to accurately grade and stage the disease prior to therapy. PMID- 16310935 TI - Role of glucocorticoids in dopamine-related neuropsychiatric disorders. AB - 'Psychoneuroendocrinology' is now quickly emerging as a hot interdisciplinary research field that addresses the interplay between neuronal and endocrine signaling in psychiatric diseases. Both glucocorticoid hormones and dopamine have an important role in maintaining normal brain functions. In this review, molecular and mechanistic aspects of glucocorticoid effects on brain function and behavior will be discussed with specific reference to dopamine signaling. PMID- 16310936 TI - The effect of grain size of rock phosphate amendment on metal immobilization in contaminated soils. AB - When rock phosphates (RP) are used to remediate Pb-contaminated soils, their effectiveness is likely affected by their grain size. In this study, the effect of grain size of rock phosphate on the effectiveness of heavy metal immobilization in two contaminated soils was measured in pot experiment. Rock phosphate was used with four different grain sizes: <35, 35-72, 72-133 and 133 266microm. The application rate of rock phosphate in two soils was determined based on P/metals (Pb, Zn, Cu and Cd) molar ratio of 5.0 in the soils. The results showed that rock phosphate of the smallest grain size (<35microm) was superior to all of other grain sizes more than 35microm for reducing uptake in plant (Brassica oleracea L.) shoots for Cd (19.6-50.0%), Pb (21.9-51.4%) and Zn (22.4-34.6%), respectively, as compared with the soil without application of rock phosphate. Sequential extraction analysis indicated that rock phosphate was most effective for soil Pb to induced transformation from non-residual fractions to a residual fraction than that for Zn and Cd. Such transformation was probably through dissolution of Pb associated with exchangeable (EX), organic fraction (OC), acidic fraction (AC) and amorphous Fe and Al oxides-bound (OX) fraction and precipitation of pyromorphite-like minerals. Results suggested that the rock phosphate with small grain size was superior to that with large grain size for in situ remediation technology. PMID- 16310937 TI - Decomposition of some polynitro arenes initiated by heat and shock Part I. 2,4,6 Trinitrotoluene. AB - Samples of 2,4,6-trinitrotoluene (TNT) exposed to heat or to shock and residues after their detonation have been analyzed chromatographically (LC-UV and LC/MS). It was found that the main identified decomposition intermediates are identical in all the three cases. 4,6-Dinitro-2,1-benzoisoxazole and 2,4,6 trinitrobenzaldehyde are the most reactive from them. It has been stated that the chemical micro-mechanism of the primary fragmentations of shock-exposed TNT molecules and/or its detonation transformation should be the same as in the case of its low-temperature thermal decomposition. PMID- 16310938 TI - Combustion of chlorinated volatile organic compounds (VOCs) using bimetallic chromium-copper supported on modified H-ZSM-5 catalyst. AB - The paper reports on the performance of chromium or/and copper supported on H-ZSM 5(Si/Al = 240) modified with silicon tetrachloride (Cr1.5/SiCl4-Z, Cu1.5/SiCl4-Z and Cr1.0Cu0.5/SiCl4-Z) as catalysts in the combustion of chlorinated VOCs (Cl VOCs). A reactor operated at a gas hourly space velocity (GHSV) of 32,000 h(-1), a temperature between 100 and 500 degrees C with 2500 ppm of dichloromethane (DCM), trichloromethane (TCM) and trichloroethylene (TCE) is used for activity studies. The deactivation study is conducted at a GHSV of 3800 h(-1), at 400 degrees C for up to 12 h with a feed concentration of 35,000 ppm. Treatment with silicon tetrachloride improves the chemical resistance of H-ZSM-5 against hydrogen chloride. TCM is more reactive compared to DCM but it produces more by products due to its high chlorine content. The stabilization of TCE is attributed to resonance effects. Water vapor increases the carbon dioxide yield through its role as hydrolysis agent forming reactive carbocations and acting as hydrogen supplying agent to suppress chlorine-transfer reactions. The deactivation of Cr1.0Cu0.5/SiCl4-Z is mainly due to the chlorination of its metal species, especially with higher Cl/H feed. Coking is limited, particularly with DCM and TCM. In accordance with the Mars-van Krevelen model, the weakening of overall metal reducibility due to chlorination leads to a loss of catalytic activity. PMID- 16310939 TI - Treatment of olive mill effluents by coagulation-flocculation-hydrogen peroxide oxidation and effect on phytotoxicity. AB - The pre-treatment of olive mill effluents (OME) by means of coagulation flocculation coupling various inorganic materials and organic poly-electrolytes was investigated. Tests were conducted with two different OME with chemical oxygen demand (COD) contents of 61.1 and 29.3 g/L, total suspended solids (TSS) of 36.7 and 52.7 g/L and total phenolic contents (TP) of 3.5 and 2.5 g/L, respectively. Inorganic materials such as lime, iron, magnesium and aluminum as well as four cationic and two anionic commercial poly-electrolytes were employed either alone or in various combinations and screened with respect to their efficiency in terms of TSS, TP and COD removal, the amount of sludge produced and the phytotoxicity of the resulting liquid to lettuce seeds. Coupling lime or ferrous sulphate (in the range of several g/L) with cationic poly-electrolytes (in the range of 200-300 mg/L) led to quantitative TSS removal, while COD and TP removal varied between about 10-40% and 30-80%, respectively, depending on the materials and the effluent in question; separation efficiency generally decreased with decreasing coagulant and/or flocculant concentration. To enhance organic matter degradation, iron-based coagulation was coupled with H(2)O(2), thus simulating a Fenton reaction and this increased COD reduction to about 60%. The original, untreated OME was strongly phytotoxic to lettuce seeds even after several dilutions with water; however, phytotoxicity decreased considerably following treatment with lime and cationic poly-electrolytes; this was attributed to the removal of phenols and other phytotoxic species from the liquid phase. PMID- 16310940 TI - Removal of lead(II) and cadmium(II) from aqueous solutions using grape stalk waste. AB - The sorption of lead and cadmium from aqueous solutions by grape stalk waste (a by-product of wine production) was investigated. The effects of the contact time, pH of the solution, ionic medium, initial metal concentration, other metal ions present and ligands were studied in batch experiments at 20 degrees C. Maximum sorption for both metals was found to occur at an initial pH of around 5.5. The equilibrium process was described well by the Langmuir isotherm model, with maximum grape stalk sorption capacities of 0.241 and 0.248 mmol g(-1) for Pb(II) and Cd(II), respectively, at pH around 5.5. Kinetic studies showed good correlation coefficients for a pseudo-second-order kinetic model. The presence of NaCl and NaClO(4) in the solution caused a reduction in Pb and Cd sorption, the latter being more strongly suppressed. The presence of other metals in the uptake process did not affect the removal of Pb, while the Cd uptake was much reduced. HCl or EDTA solutions were able to desorb lead from the grape stalks completely, while an approximately 65% desorption yield was obtained for cadmium. From the results obtained it seems that other mechanisms, such as surface complexation and electrostatic interactions, must be involved in the metal sorption in addition to ion exchange. PMID- 16310941 TI - Selective removal of lead and bromide from a hazardous industrial solid waste using Limited Acid Demand and Separation Factor at ambient conditions. AB - A detailed methodology is described for the selective removal of lead and bromide content from an industrial toxic solid waste, containing also large quantities of iron. Limited Acid Demand (LAD) was examined in order to avoid the co-dissolution of undesirable constituents. The Separation Factor (SF), which is the concentration ratio of dissolved constituents in the resulting leaching solution after the application of extraction stage, can describe the selective release of lead against iron, according to the variation of major leaching parameters (i.e. the molarity of used acid and the liquid to solid (L/S) ratio). A general equation, describing the overall leaching process was obtained, containing a specific "inhibition constant" and indicating that when the initial stoichiometry of leaching procedure and the applied L/S ratio are known, then the removal of lead can be predicted. PMID- 16310942 TI - Comparative sorption kinetic studies of ammonium onto zeolite. AB - The sorption kinetics of ammonium onto three types of zeolite, natural zeolite, natural zeolite covered by biofilm and ammonium-bearing zeolite covered by biofilm, at two particle sizes were studied. The pseudo-first order and pseudo second order models were fitted to the results by a non-linear method. The batch sorption model, based on a pseudo-second order mechanism, was applied to predict the rate constant of sorption, the equilibrium capacity and the initial sorption rate. Ion exchange between NH(4)(+) and cations in the zeolite increased with decreasing particle size of the zeolite. Biofilm covered on the zeolite did not affect the ion exchange for the smaller particle size but decreased the ion exchange capacity by 22% for the larger particle size. In addition, bioregeneration should be considered for the recovery of ion exchange capacity of the bio-zeolite made from ammonium-bearing zeolite by the microorganisms in the biofilm, 78.0 and 63.9% regeneration for the smaller and larger particle size of zeolite, respectively. PMID- 16310943 TI - Harnessing data mining to explore incident databases. AB - Large numbers of incident related databases have been established in the last three decades. The majority of attempts to explore these data marts were trials to identify patterns via first glance into the datasets. This study investigated a subset of incidents from fixed facilities in Harris County, TX, extracted from the National Response Center database. By classifying the information into groups and using data mining techniques, interesting patterns of incidents according to characteristics such as type of equipment involved, type of chemical released and causes involved were revealed and further these were used to modify the annual failure probabilities of equipments. PMID- 16310944 TI - Effect of organic additives on the mitigation of volatility of 1-nitro-3,3' dinitroazetidine (TNAZ): next generation powerful melt cast able high energy material. AB - 1-Nitro-3,3'-dinitroazetidine (TNAZ) was synthesized based on the lines of reported method. Thermolysis studies on synthesized and characterized TNAZ using differential scanning calorimetry (DSC) and hyphenated TG-FT-IR techniques were undertaken to generate data on decomposition pattern. FT-IR of decomposition products of TNAZ revealed the evolution of oxides of nitrogen and HCN containing species suggesting the cleavage of C/N-NO(2) bond accompanied with the collapse of ring structure. The effect of incorporation of 15% additives namely, 3-amino 1,2,4-triazole (AT), 3,5-diamino-1,2,4-triazole (DAT), carbohydrazide (CHZ), 5,7 dinitrobenzofuroxan (DNBF), bis (2,2-dinitropropyl) succinate (BNPS), triaminoguanidinium nitrate (TAGN), 2,4,6-trinitrobenzoic acid (TNBA) and nitroguanidine (NQ) on the volatility of TNAZ was investigated by undertaking thermogravimetric analysis. The TG pattern brings out the potential of BNPS and TAGN as additives to mitigate the volatility of TNAZ. The influence of additives on thermal decomposition of pattern of TNAZ was also investigated by DSC. The DSC results indicated that the additives did not have appreciable effect on the melting point of TNAZ. Scanning electron microscopic (SEM) studies were carried out to investigate the effect of additives on morphology of TNAZ. This paper also discusses the possible mechanism involved in between the TNAZ and TAGN and BNPS. It appears that the formation of charge transfer complex formation between the TNAZ and TAGN/BNPS. The effect of addition of high explosives such as CL-20, HMX and RDX on thermo-physical characteristics of TNAZ is also reported in this paper. PMID- 16310945 TI - Kinetic study on photocatalytic degradation of C.I. Acid Yellow 23 by ZnO photocatalyst. AB - The potential of a common semiconductor, ZnO, has been explored as an effective catalyst for the photodegradation of C.I. Acid Yellow 23 (AY23). The effects of process parameters such as, catalyst loading, initial dye concentration, light intensity, and pH on the extent of photodegradation have been investigated. Substantial reduction of COD, besides removal of color, was also achieved. A rate equation for the degradation based on Langmuir-Hinshelwood (L-H) model has been proposed. The results show that the adsorption constant (K(ads)) and rate constant (k(L-H)) in L-H model are dependent to the light intensity, and increase with increasing the light intensity. With inserting the light intensity parameter to L-H equation, this model can be used for predicting the removal rate at different light intensities and initial concentrations of AY23. A comparison between experimental and calculated apparent reaction rate constants shows that the results obtained from the L-H modified model are in good agreement with experimental data. PMID- 16310946 TI - Recycling spent zinc manganese dioxide batteries through synthesizing Zn-Mn ferrite magnetic materials. AB - A novel process to reclaim spent zinc manganese dioxide batteries (SDBs) through synthesizing Zn-Mn ferrite magnetic materials is present. Firstly, the dismantling, watering, magnetism, baking and griddling steps were consecutively carried out to obtain iron battery shells, zinc grains and manganese compounds using the collected SDBs, and then these separated substances were dissolved with 4 mol L(-1) H(2)SO(4) to prepare FeSO(4), ZnSO(4) and MnSO(4) reactant solutions. Secondly, Zn-Mn ferrites with stoichiometric ratio of Mn(0.26)Zn(0.24)FeO(2) were synthesized using chemical co precipitation process with ammonium oxalate precipitator. The XRD results showed that the obtained Zn-Mn ferrites had spinel structure and high purity at the calcining temperatures of 850-1250 degrees C. With the increase of calcining temperature, the finer crystalline structure could be formed, and their intensity of saturation magnetization reached the highest value at 1150 degrees C. The magnetization performances of Zn-Mn ferrites prepared from the SDBs were similar to that of from analysis reagents, suggesting the feasibility to synthesize Zn-Mn ferrites with high properties from SDBs. PMID- 16310947 TI - Coal ash conversion into effective adsorbents for removal of heavy metals and dyes from wastewater. AB - Fly ash was modified by hydrothermal treatment using NaOH solutions under various conditions for zeolite synthesis. The XRD patterns are presented. The results indicated that the samples obtained after treatment are much different. The XRD profiles revealed a number of new reflexes, suggesting a phase transformation probably occurred. Both heat treatment and chemical treatment increased the surface area and pore volume. It was found that zeolite P would be formed at the conditions of higher NaOH concentration and temperature. The treated fly ash was tested for adsorption of heavy metal ions and dyes in aqueous solution. It was shown that fly ash and the modified forms could effectively absorb heavy metals and methylene blue but not effectively adsorb rhodamine B. Modifying fly ash with NaOH solution would significantly enhance the adsorption capacity depending on the treatment temperature, time, and base concentration. The adsorption capacity of methylene blue would increases with pH of the dye solution and the sorption capacity of FA-NaOH could reach 5 x 10(-5) mol/g. The adsorption isotherm could be described by the Langmuir and Freundlich isotherm equations. Removal of copper and nickel ions could also be achieved on those treated fly ash. The removal efficiency for copper and nickel ions could be from 30% to 90% depending on the initial concentrations. The increase in adsorption temperature will enhance the adsorption efficiency for both heavy metals. The pseudo second-order kinetics would be better for fitting the dynamic adsorption of Cu and Ni ions. PMID- 16310948 TI - Consequences of major accidents: assessing the number of injured people. AB - Quantitative risk assessment studies of accident scenarios usually involve estimating the number of fatalities that can be expected. The number of people injured, however, is seldom evaluated because it implies significant additional effort and often the information required to perform this evaluation is not available. However, the number of injured people can be very important for emergency planning, especially in relatively large accidents. In this paper, a set of 975 accidents were selected for analysis, with the aim of searching for a relationship between the number of people killed and the number of people injured. As the data were scattered, principal component analysis and clustering analysis were applied to identify the data subsets that could undergo a selective, specific statistical treatment. Further treatment of these subsets led to mathematical expressions that are used to estimate the probable number of injured people as a function of the number of fatalities for all accidents, as well as for gas cloud, fire and explosion events, respectively. PMID- 16310949 TI - Aqueous Cr(VI) photo-reduction catalyzed by TiO2 and sulfated TiO2. AB - TiO(2) and sulfated TiO(2) (SO(4)(2-)/TiO(2)) catalysts with different textural properties were prepared under different calcination temperatures and the photo reduction of Cr(VI) to Cr(III) catalyzed by these catalysts was investigated. For the photocatalytic reduction of Cr(VI), the photocatalytic activities of the TiO(2) samples were found to be strongly dependent of the calcination temperature and TiO(2) calcined at 400 degrees C showed a higher catalytic activity compared to other TiO(2) catalysts. In contrast, sulfation of TiO(2) stabilized the catalytic activities of SO(4)(2-)/TiO(2) catalysts. At low calcination temperature, SO(4)(2-)/TiO(2) catalysts exhibited catalytic activities almost comparable with that of TiO(2) and the catalytic activities of SO(4)(2-)/TiO(2) catalysts were markedly higher than TiO(2) under high calcination temperature. In addition, the removal of surface SO(4)(2-) of SO(4)(2-)/TiO(2) catalyst led to a marked decrease of the catalytic activity for Cr(VI) photo-reduction, suggesting that the presence of surface SO(4)(2-) provided an acid environment over the catalyst surface and favored the photo-reduction of Cr(VI). PMID- 16310950 TI - Biosorption of lead, copper and cadmium by an indigenous isolate Enterobacter sp. J1 possessing high heavy-metal resistance. AB - This study was undertaken to investigate biosorption kinetics and equilibria of lead (Pb), copper (Cu) and cadmium (Cd) ions using the biomass of Enterobacter sp. J1 isolated from a local industry wastewater treatment plant. Efficiency of metal ion recovery from metal-loaded biomass to regenerate the biosorbent was also determined. The results show that Enterobacter sp. J1 was able to uptake over 50mg of Pb per gram of dry cell, while having equilibrium adsorption capacities of 32.5 and 46.2mg/g dry cell for Cu and Cd, respectively. In general, Langmuir and Freundlich models were able to describe biosorption isotherm fairly well, except that prediction of Pb adsorption was relatively poor with Langmuir model, suggesting a different mechanism for Pb biosorption. Adjusting the pH value to 3.0 led to nearly complete desorption of Cd from metal-loaded biomass, while over 90% recovery of Pb and Cu ions was obtained at pH300 genes encoded by chromosome 21 with specific phenotypic features is a goal that becomes more feasible with the increasing availability of large scale functional, expression and evolutionary data. These data are dispersed among diverse databases, and the variety of formats and locations, plus their often rapid growth, makes access and assimilation a daunting task. To aid the Down syndrome and chromosome 21 community, and researchers interested in the study of any chromosome 21 gene or ortholog, we are developing a comprehensive chromosome 21-specific database with the goals of (i) data consolidation, (ii) accuracy and completeness through expert curation, and (iii) facilitation of novel hypothesis generation. Here we describe the current status of data collection and the immediate future plans for this first human chromosome specific database. PMID- 16310978 TI - Communicating risk and uncertainty in relation to development and implementation of disease control policies. AB - Scientific evidence is one of the key factors to be considered in the development of disease control policies. It is generated using investigations into cause effect relationships, which usually produce results that are associated with a varying degree of uncertainty. Experience has shown that taking account of these uncertainties can become a formidable challenge for policy makers when devising the strategies and when communicating them to stakeholders. The situation has been further complicated by a reduction in public trust in scientific evidence. It is now recognised that this challenge cannot be managed by simply providing more information, but it is also necessary to consider the influence that variation in risk perception amongst stakeholders has on their response to and commitment towards the policies. PMID- 16310979 TI - The pathogenesis and pathology of bovine tuberculosis with insights from studies of tuberculosis in humans and laboratory animal models. AB - This paper reviews key insights the discipline of pathology has contributed to our understanding of bovine tuberculosis in the context of findings of studies of tuberculosis in humans and laboratory animal models. Analysis and extrapolation of data from other species have the potential to expand our understanding of the pathogenesis of the disease in cattle. The distribution of lesions in affected cattle, humans and laboratory animals illustrate the primacy of the respiratory tract as portal of infection and raise questions about the role of the upper respiratory tract surface, tonsil and dorsal lung regions in disease pathogenesis and transmission. The mechanisms behind significant pathological processes such as necrosis, apoptosis and liquefaction, occurring within lesions, are explored and their potential practical significance assessed in the context of herd disease dynamics and vaccine development. It is proposed that effective 'innate' host defences result in many animals and humans remaining disease-free and tuberculin test negative following exposure to infection. Furthermore, the concepts of latency and disease reactivation, considered significant factors in perpetuating tuberculosis in human populations, are explored in the context of the bovine disease. PMID- 16310980 TI - An update on bovine tuberculosis programmes in Latin American and Caribbean countries. AB - Of the approximately 374 million cattle in Latin America and the Caribbean, 70% are held in areas where rates of Mycobacterium bovis infection in cattle are higher than 1%. The remaining 30% are in countries where infection affects less than 1% of cattle, including 62 million in countries where bovine tuberculosis infection is virtually nil. Measures for controlling bovine tuberculosis are partially or extensively applied in most of the countries in the Region. These measures are based on test and slaughter, notification, post-mortem inspection and surveillance in slaughterhouses. A coordinated production, standardization and quality control of purified protein derivatives is urgently required for use in control and eradication campaigns in order to assure reliability of reagents and comparability of data on tuberculin testing within the Region. On the basis of information from Argentina, M. bovis is estimated to cause 2% of all human cases of tuberculosis in the Region. Slaughterhouse and dairy farms workers are most-frequently infected, with infection occurring via the respiratory tract. Various in vitro assays for the diagnosis of bovine tuberculosis have been developed and/or assessed in the Region, and DNA fingerprinting has been applied for a comprehensive understanding of the epidemiology of bovine tuberculosis at the local and regional level. PMID- 16310981 TI - A long-term study in Angora goats experimentally infected with Mycobacterium avium subsp. paratuberculosis: clinical disease, faecal culture and immunological studies. AB - Two longitudinal experiments involving Angora goats challenged with either bovine or ovine strains of Mycobacterium avium subspecies paratuberculosis (Map) have been conducted over a period of 54 and 35 months, respectively. Blood samples for the interferon-gamma (IFN-gamma) test and the absorbed ELISA and faecal samples for bacteriological culture were taken pre-challenge and monthly post-challenge. Persistent shedding, IFN-gamma production, seroconversion and clinical disease occurred earlier with the bovine Map gut mucosal tissue challenge inoculum than with cultured bacteria. The IFN-gamma responses of the gut mucosal tissue and bacterial challenge groups were substantially and consistently higher than those of the control group. The in vivo and cultured cattle strains were much more pathogenic for goats than the sheep strains with persistent faecal shedding, seroconversion and clinical disease occurring in the majority of bovine Map challenged goats. With the ovine Map, 3 goats developed persistent antibody responses but only one of these goats developed persistent faecal shedding and clinical disease. However, there was no significant difference between the IFN gamma responses of the tissue challenged, bacterial challenged and control groups. Compared with sheep, the ELISA appeared to have higher sensitivity and the IFN-gamma test lower specificity. PMID- 16310982 TI - The cytolethal distending toxin among Helicobacter pullorum strains from human and poultry origin. AB - Helicobacter pullorum has been associated with diarrhoea, gastroenteritis and liver disease in humans and with hepatitis and enteritis in poultry. The purpose of the present study was to examine whether cytolethal distending toxin was present among 10 poultry and three human H. pullorum isolates and whether a different level of cytolethal distending toxin activity was noted. A PCR assay was performed to detect the cdtB gene. In addition, epithelial Hep-2 cells inoculated with sonicate from all strains were observed microscopically and DNA analysis of these cells was done by flow cytometry. All H. pullorum isolates harboured the cdtB gene, but functional cytolethal distending toxin activity was only demonstrated in the human H. pullorum strain CCUG 33839. A significant number of cells treated with sonicate from this strain were enlarged. The nuclei were distended proportionally. Giant cells and multinucleated cells were observed as well. In addition, stress fibers accumulated. DNA analysis by flow cytometry revealed 31.0% of these cells at the S/G2 stage of the cell cycle. The tested poultry and human H. pullorum isolates all possess the cdtB gene, but under the circumstances adopted in this study only the human strain CCUG 33839 seems to show biological activity typically for CDT in vitro. PMID- 16310983 TI - Role of SPI-1 in the interactions of Salmonella Typhimurium with porcine macrophages. AB - Salmonella Pathogenicity Island 1 (SPI-1) genes are indispensable for virulence of Salmonella Typhimurium in mice after oral challenge. These genes mediate invasion in intestinal epithelial cells and induce cell death in murine macrophages. The role of SPI-1 in the pathogenesis of Salmonella Typhimurium infections in food producing animals is not known. It was the aim of the present study to characterize the interactions of a porcine Salmonella Typhimurium field strain and its isogenic mutants in the SPI-1 genes hilA, sipA and sipB with porcine macrophages. SPI-1 was found to be important in the invasion of porcine pulmonary alveolar macrophages (PAM) and the induction of the formation of spacious phagosomes. Both early and delayed cytotoxicity were seen in PAM, but only the early cytotoxicity was SPI-1 dependent. Exposure of PAM to Salmonella Typhimurium induced the production of reactive oxygen species (ROS) and interleukin-8, but no differences were noticed between the induction mediated by the wild type strain and its SPI-1 mutant strains. In conclusion, invasion of porcine macrophages and the induction of early, but not delayed, cytotoxicity by Salmonella Typhimurium is SPI-1 dependent. SPI-1 dependent invasion, however, is not a prerequisite to induce a pro-inflammatory response. PMID- 16310984 TI - Genetic basis of toxic reactions to drugs and chemicals. AB - Inter-individual drug effects are subject to substantial variability. There are multiple reasons based on pathophysiological factors and environmental interactions, but also genetic characteristics. Groundbreaking successes have been achieved in the field of pharmacogenomics and toxicogenomics. In particular, the identification of hereditary polymorphisms in genes of the cytochrome P450 system and phase II-enzymes such as TMPT contributed considerably to the explanation of the individually varying pharmacokinetics of a number of drugs. Furthermore, hereditary variations in genes of membrane drug transporters were recently discovered. Along with these factors, which could influence pharmacokinetics, strong efforts have been undertaken to clarify the role of genetic polymorphisms in receptors or signal transduction proteins modulating drug efficacy. Particularly for malignant diseases such as bladder or lung cancer, polymorphic foreign compound metabolizing enzymes have been identified as susceptibility factors, modulating an individual's cancer risk dependent on the extent of environmental exposure. This review focuses on the role of the polymorphic phase I enzymes cytochrome P450 1A1, 1A2, 1B1, 2C9, 2C19, 2D6, 3A5 and myeloperoxidase as well as on the phase II-enzymes arylamine N acetyltransferases 1 and 2, glutathione S-transferases M1 and T1, and thiopurine S-methyltranferases as detoxifying but also toxifying factors, modulating pharmacokinetics and disease susceptibility. PMID- 16310985 TI - Damage of zinc fingers in DNA repair proteins, a novel molecular mechanism in carcinogenesis. AB - Zinc finger motifs participate in protein-nucleic acid and protein-protein interactions in many groups of proteins, including those involved in DNA repair. The Zn(II) ion, bonded tetrahedrally to cysteine thiolates and/or histidine imidazole groups, maintains the three-dimensional structure, crucial for the function of the domain. Zinc fingers can thus be compromised by a substitution of Zn(II) with another metal ion or by a release of Zn(II), due to the oxidation of thiolate donors. The latter may result from an action of redox-active metals or other oxidative agents. Studies in cell cultures and ex vivo demonstrated that soluble compounds of definite carcinogenic metals and metalloids, such as arsenic, cadmium and nickel, and putative carcinogens, including cobalt and lead, inhibit zinc finger containing DNA repair proteins. Further experiments demonstrated that these metals, as well as endogeneous oxidative substances, including hydrogen peroxide, nitrosoglutathione, and reducible selenium compounds damage or distort zinc finger domains. This reactivity can therefore be regarded as a novel molecular mechanism in carcinogenesis. PMID- 16310986 TI - Controlling interrelationships of progesterone/LH and estradiol/LH in mares. AB - The interrelationships of progesterone, estradiol, and LH were studied in mares (n=9), beginning at the first ovulation (Day 0) of an interovulatory interval. An increase in mean progesterone concentrations began on Day 0 and reached maximum on Day 6, with luteolysis beginning on Day 14. A common progesterone threshold concentration of about 2 ng/ml for a negative effect on LH occurred at the beginning and end of the luteal phase. Progesterone and LH concentrations decreased at a similar rate from Day 6 until the onset of luteolysis on Day 14, consistent with a decreasing positive effect of LH on progesterone. Concentrations of LH during the increase in the ovulatory surge consisted of two linear regression segments involving a rate of 0.4 ng/ml/day for Days 14-22 and 1.8 ng/ml/day for Day 22 to 1 day after the second ovulation. The end of the first segment and beginning of the second segment was 2 days before ovulation and was the day the ovulatory estradiol surge was at a peak. PMID- 16310987 TI - Nuclear and microtubule remodeling and in vitro development of nuclear transferred cat oocytes with skin fibroblasts of the domestic cat (Felis silvestris catus) and leopard cat (Prionailurus bengalensis). AB - The leopard cat (Prionailurus bengalensis), a member of the felidae family, is a threatened animal in South Korea. In terms of protecting endangered felids, nuclear transfer (NT) is a potentially valuable technique for assuring the continuation of species with dwindling numbers. In the present experiment, nuclear and microtubule remodeling and the in vitro developmental potential of enucleated domestic cat oocytes reconstructed with nuclei of somatic cells from either domestic cat fibroblast (DCF) or leopard cat fibroblast (LCF) were evaluated. Microtubule aster is allocated to de-condensed chromatin following nuclear transfer (3h after activation) of fibroblast cells from both domestic and leopard cats, suggesting the introduction of a somatic cell centrosome. The transferred fibroblast nuclei formed a large, swollen, pronuclear-like structure in most reconstructed oocytes, in the cat or leopard cat. At 18h following nuclear transfer, mitosis occurred, and according to the photo (F) it appears that spindle microtubules and two asters were observed. The percentages of blastocyst formation from nuclear transfer embryos derived from domestic cat fibroblasts (4/46, 8.6%) were not significantly different than those for nuclear transfer embryos constructed with leopard cat fibroblasts (4/52, 7.6%). These results indicate that nuclear and microtubule remodeling processes and in vitro developmental ability are similar in reconstructed cat oocytes following transfer of nuclei from either domestic or leopard cats. PMID- 16310988 TI - Hydroxyethylstarch microcapsules: a preliminary study for tumor immunotherapy application. AB - The objective of this work was to prepare microcapsules which would allow protection and slow release of antigens used for melanoma immunotherapy treatment. Hydroxyethylstarch (HES) microcapsules were prepared using interfacial cross-linking with terephthaloyl chloride (TC). They were characterized with respect to morphology (microscopy) and size (in the 4-15 microm range). Bovine serum albumin (BSA) was used as model protein for loading and release studies. Microcapsules were loaded with solutions at different protein concentrations (0.5 5%). The maximum loading efficiency (20%) was observed with the concentration of 2.5%, which allowed a loading capacity near 100%. Confocal laser scanning microscopy (CLSM) visualization showed that BSA was entrapped within the microcapsules and not only associated to their outer surface. BSA-release studies showed a 20% BSA release within 30 min while 80% remained entrapped in the microcapsules for 4 days. Microcapsules were degraded by alpha-amylase and addition of esterase to alpha-amylase enhanced slightly their degradation. In vitro studies on melanoma cells showed that HES microcapsules were non-toxic. Preliminary in vivo studies demonstrated that microcapsules were biodegradable after intraperitoneal injection (i.p.). The observation of peritoneal wash showed a complete degradation within 7 days, indicating a possible application as an in vivo drug delivery system especially to enhance the presentation of antigens. PMID- 16310989 TI - Lung surfactant as a drug delivery system. AB - Lung surfactant is a complex mixture of mainly phospholipids and proteins. The composition leads to a unique spreading effect of the surfactant as well as spontaneous vesicle formation, which may be favourable characteristics of a drug delivery system for pulmonary delivery. The aim of study was to investigate the potential use of the surfactant extract, HL10 (LeoPharma, DK) as a drug delivery system. Studies involved incorporation of hydrophilic- and amphipathic model drugs (sucrose and acylated peptides) into HL10 and elucidation of the influence of surfactant proteins on the HL10 behaviour. Results showed that HL10 vesicles did not retain sucrose indicating formation of leaky vesicles. Studying the influence of surfactant proteins on release from DPPC-liposomes showed tendencies toward a protein-induced release. Hence, the surfactant proteins may influence the membrane lipid packing and characteristics resulting in leakiness of the membranes. Incorporation of acylated peptides into HL10 depended on the chain length rendering a successful incorporation of the peptide acylated with C14-acyl chains. This study suggests that HL10 may be a promising drug delivery system for the pulmonary delivery of amphipathic drug substances, e.g. therapeutically active acylated peptides (e.g. acylated insulin). PMID- 16310990 TI - Biodegradable microparticulate system of captopril. AB - Albumin microparticles have found many applications in diagnosis and treatment in recent years and more than 100 diagnostic agents and drugs have been incorporated into albumin microparticles. In the present study, bovine serum albumin (BSA) based microparticles bearing captopril were prepared by an emulsification-heat stabilization technique. Four batches of microparticles with varying ratio of drug and polymer were prepared. The prepared microparticles were studied for drug loading, particle size distribution, in vitro release characteristics, in vivo tissue distribution study and stability studies. The microparticles had mean diameter between 2 and 11 microm of which more than 70% were below 5 microm and incorporation efficiency of 41-63% was obtained. In vitro release profile for formulations containing captopril-loaded albumin microparticles with heat stabilizing technique shows slow controlled release up to 24 h. The in vivo result of drug-loaded microparticles showed preferential drug targeting to liver followed by lungs, kidneys and spleen. Stability studies showed that maximum drug content and closest in vitro release to initial data were found in the formulation stored at 4 degrees C. In the present study, captopril-loaded BSA microparticles were prepared and targeted to various organs to a satisfactory level and were found to be stable at 4 degrees C. PMID- 16310991 TI - Cutaneous metabolism of a dipeptide influences the iontophoretic flux of a concomitant uncharged permeant. AB - Passive and iontophoretic transport of the model dipeptide tyrosine-phenylalanine (TyrPhe) that is subject to cutaneous metabolism and the uncharged glucose derivative benzyl-2-acetamido-2-deoxy-alpha-d-glucopyranoside (BAd-alpha-Glc) used as electroosmosis marker through heat-separated human epidermis was investigated in vitro. TyrPhe and BAd-alpha-Glc were used separately and in combination in order to determine their interaction in terms of permeability and the influence of skin metabolism of TyrPhe on permeation rate and tissue retention of itself and of BAd-alpha-Glc. TyrPhe was chemically and electrochemically stable but underwent considerable degradation in the epidermis under reflection boundary conditions with generation of degradation products tyrosine (Tyr) and phenylalanine (Phe) confirming cutaneous metabolism of TyrPhe in heat-separated human epidermis, which was more pronounced at pH 4.5 than at pH 3.0. As a result, no reproducible epidermis permeation of TyrPhe at pH 3 and no permeation at all at pH 4.5 was measured regardless of the presence of BAd-alpha Glc, accompanied by increased levels of Tyr and Phe compared to blank runs. Low temperature (4 degrees C) at both pH values and addition of o-phenanthroline at pH 3 but not at pH 4.5 yielded reproducible TyrPhe permeation and blank, i.e., endogenous levels of Tyr and Phe evidencing inhibition of degradation. Constant voltage anodal iontophoresis marginally reduced BAd-alpha-Glc flux at pH 3 and 4.5 compared to the passive flux. In combination with TyrPhe, iontophoretic flux of BAd-alpha-Glc was increased markedly compared to the passive one when TyrPhe was metabolized in the tissue, while no such increase was observed when TyrPhe metabolism was inhibited. The increase of BAd-alpha-Glc iontophoretic flux was accompanied by a considerable decrease of the BAd-alpha-Glc amount retained in the epidermis. The presence of the generated Tyr and Phe, therefore, appears to be related to a decrease of the BAd-alpha-Glc amount retained in the epidermis upon application of an electrical voltage and an enhancement of its iontophoretic flux. Thus, an interaction between the concurrent permeants at the level of tissue retention induced by metabolism can influence the apparent iontophoretic permeation. PMID- 16310992 TI - Are chitosan formulations mucoadhesive in the human small intestine? An evaluation based on gamma scintigraphy. AB - Rapid passage through the proximal intestine can result in the low bioavailability of a drug substance with site-specific absorption characteristics in the upper gastrointestinal tract. To overcome this, there is increasing interest in developing gastro-retentive formulations and/or formulations that linger in the proximal parts of the small intestine, e.g. by using mucoadhesive polymers as excipients in formulations. In our recent study, we used neutron activation-based gamma scintigraphy to evaluate the gastro-retentive properties of formulations containing chitosan (Mw 150 kDa) in man. At the same time, we had an opportunity to monitor the transit of the formulations (40 or 95% of chitosan) in the small intestine. Gamma scintigraphic investigations revealed that although the chitosan studied had exhibited marked mucoadhesive capacities in vitro, retention of the chitosan formulations in the upper gastrointestinal tract was not sufficiently reproducible and the duration of retention was relatively short. In 3 volunteers out of 10, the formulation adhered to the gastric mucosa (retention times varied from 1.25 to 2.5 h) and in two volunteers to the upper small intestine (approximate retention time 45 min). In one case, the formulation adhered to the oesophagus. The system failed to increase the bioavailability of furosemide, a drug site-specifically absorbed in the upper gastrointestinal tract. As far as the kind of formulation studied is concerned, preparation of a system that is site-specific to the stomach and/or the upper small intestine seems difficult if the proposed mechanism of action is mucoadhesion. The results suggest that other mechanisms of action should also be studied. PMID- 16310993 TI - Effects of three purgative decoctions on inflammatory mediators. AB - In traditional Chinese medicine (TCM), there are three Cheng-Chi-Tang decoctions (CCTDs) including: Ta-Cheng-Chi-Tang (TCCT), Xiao-Chen-Chi-Tang (XCCT) and Tiao Wei-Chen-Chi-Tang (TWCCT), which are the frequently used purgative remedies to treat "internal heat"-induced symptoms like a bloated and painful abdomen, hard stools and fever, etc. Constituents in each formulation are Rheum palmatum L. (Polygonaceae), Magnolia officinalis Rehd. et Wils. (Magnoliaceae), Citrus aurantium L. (Rutaceae), Mirabilitum (mirabilite, crystals of sodium sulfate, Na2SO4) for TCCT; Rheum palmatum, Magnolia officinalis, Citrus aurantium for XCCT; and Rheum palmatum, Mirabilitum, Glycyrrhiza uralensis Fisch. (Leguminosae) for TWCCT. However, the underlying mechanisms for purging internal pathological heat are far from fully clarified, and few scientific investigations have been carried out to delineate the relationships between the anti-inflammatory effects and laxative potencies of these formulations. In this study, the anti inflammatory effects of the three CCTDs on lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E (PGE2) production in RAW 264.7 cells, carrageenan-induced paw edema in mice and the laxative effect in mice were explored. The results showed that TCCT inhibited LPS-induced NO and PGE2 production and inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells more effectively than did XCCT or TWCCT. Moreover, paw edema of carrageenan treated mice was significantly attenuated in mice pretreated with 1 g/kg TCCT. TCCT also showed the strongest purgative activity among the three formulations. These findings indicate that TCCT has anti-inflammatory effects in addition to its traditionally known purgative activities. It may have potential to treat inflammatory disease conditions. PMID- 16310994 TI - Anti-inflammatory and free radical scavenging activity of Ricinus communis root extract. AB - Anti-inflammatory and free radical scavenging activities of the methanolic extract of Ricinus communis (RCM) (Euphorbiaceae) Linn. root was studied in Wistar albino rats. The methanolic extract at doses 250 and 500 mg/kg p.o. exhibited significant (P<0.001) anti-inflammatory activity in carrageenan-induced hind paw edema model. The extract at the dose of 500 mg/kg p.o. also exhibited significant (P<0.001) anti-inflammatory activity in cotton pellet granuloma model. The methanolic extract showed significant free radical scavenging activity by inhibiting lipid peroxidation initiated by carbon tetrachloride and ferrous sulphate in rat liver and kidney homogenates. The extract enhanced free radical scavenging activity of stable radical 2,2-diphenyl-1-picryl-hydrazyl (DPPH*), nitric oxide and hydroxyl radical in in vitro assay methods. The results of the study indicate that the methanolic extract of Ricinus communis root possess significant anti-inflammatory activity in acute and chronic inflammatory models in rats. The observed pharmacological activity may be due to the presence of phytochemicals like flavonoids, alkaloids and tannins present in the plant extract with various biological activities. PMID- 16310995 TI - A new antibacterial clerodane diterpenoid from the resinous exudate of Haplopappus uncinatus. AB - The antibacterial activity of Haplopappus uncinatus is attributed to a new clerodane diterpenoid, 18-acetoxy-cis-cleroda-3-en-15-oic acid (10 betaH, 16 xi 19 beta, 17beta, 20 alpha form) (1), isolated as a major component from the resinous exudates of its twigs and leaves, together with the inactive 3,5 dihydroxy-6,7,3',4'-tetramethoxyflavone, n-alkanes and a few sesquiterpenoids. PMID- 16310996 TI - Validation and casework testing of the BioPlex-11 for STR typing of telogen hair roots. AB - A new STR typing strategy has been developed allowing the simultaneous amplification and subsequent analysis of 11 polymorphic systems with amplicon sizes smaller than 270bp. The multiplex amplification reaction includes six STR loci from the European standard set of loci (ESS) for DNA databases (D3S1358, D8S1179, D21S11, THO1, FGA and VWA) as well as four additional STR systems selected for their robustness (D2S1338, D12S391, TPOX and D5S818) together with the sex-specific locus amelogenin. After PCR amplification, the multiplex reaction is splitted into two sets of STR multiplexes by using biotin labelled primers only for one set. Using streptavidin-coated Sepharose beads five STR systems are separated from the other six systems prior to being analysed in two different runs on a capillary gel electrophoresis instrument. The multiplex system was developed and tested especially for the use in forensic casework if only limited amounts or highly degraded DNA is available, for instance, when isolated from telogen hair roots. PMID- 16310997 TI - Preservation of the auditory nerve function after translabyrinthine removal of vestibular schwannoma. AB - OBJECTIVE: To evaluate the function of the postoperative auditory nerve preserved after translabyrinthine (TL) vestibular schwannoma (VS) removal. METHODS: Fifteen patients, who underwent unilateral VS resection via a TL approach, were preserved auditory nerve anatomically. The size and location of VS were measured on MRI preoperatively. After surgery, the electrical stimulation test (EST) or electrically evoked auditory brainstem response (EABR) was performed. RESULTS: Four cases (27%) out of fifteen patients were retained the functional integrity of the auditory nerve after surgery. The maximum tumor size in the group with a positive response to EST or EABR was significantly smaller than that in the group with a negative response to EST. It seems to be difficult to preserve auditory nerve function in cases where a tumor extends to the fundus of internal auditory canal. CONCLUSION: These results suggest that size of tumor and/or extension of tumor to the fundus might be important factors in preserving the auditory nerve function even if using a TL approach. PMID- 16310998 TI - A case of angiofibroma originating from the inferior nasal turbinate. AB - Angiofibromas are nasopharyngeal tumors mostly seen in adolescent males. They account for less than 0.5% of all head and neck neoplasms. Extranasopharyngeal angiofibromas, although rare, have been reported. We report a case of extranasopharyngeal angiofibroma arising from the inferior turbinate of a 62-year old male. The tumor was excised using the Denker approach. The patient is currently free of relapse 10 months after surgery. Previous extranasopharyngeal angiofibromas research was reviewed. PMID- 16310999 TI - Development of behavioural dysfunctions in accelerated-senescence OXYS rats is associated with early postnatal alterations in brain phosphate metabolism. AB - We recently demonstrated that senescence-accelerated OXYS rats may represent a model adequate for studying aging processes. The animals of this strain have a particularly short life span; they display early cataract, macular dystrophy, hypertension, and changes in cognitive and emotional spheres. Mitochondrial dysfunctions have been suggested to be a causal factor for the accelerated senescence in these animals. In the present study we investigated whether behavioral alterations in OXYS rats could be associated with changes in high energy phosphate and phospholipid metabolism of the brain in the early postnatal period. The development of behavioral dysfunction in OXYS rats relative to Wistar was investigated by measuring locomotor exploratory activity and the degree of anxiety at 4 and 12 weeks of age in open-field and elevated plus maze tests, respectively. Brain energy metabolism was evaluated at 2, 3, 4 and 12 weeks of age by calculating the ratios of PCr/Pi, PCr/ATP and phosphate potential (ATP/ADPxPi) measured by 31P NMR spectroscopy. We found that the behavioral alterations in OXYS rats, i.e. the increased anxiety and the decreased exploratory activity, were not congenital but developed during the period from 4 to 12 weeks of age. The study of high-energy phosphates has not revealed any signs of energy deficiency in OXYS rat's brain but indicated changes in PCr metabolism at 2 and 3 weeks of age when compared with Wistar rats. Furthermore, alterations in phospholipids turnover were also found in young OXYS rats. The data suggest that the changes in phosphate metabolism may have impacts on the development of behavioral deficits in OXYS rats. PMID- 16311000 TI - Nuclear flow FISH: isolation of cell nuclei improves the determination of telomere lengths. AB - Understanding telomere biology is of utmost importance for aging and cancer research. An essential tool is the determination of telomere length, which traditionally is done by telomere restriction fragment analysis, a laborious and time consuming method. Therefore, large efforts have been made to establish alternative methods like flow FISH analysis. This method, combining fluorescence in situ hybridization with a telomere specific peptide nucleic acid probe and flow cytometry, measures single cells, is suitable for analysis of non-dividing cells, and can be performed within 24 h. However, when performing flow FISH analysis with normal human kidney epithelial cells, we observed strong variation of autofluorescence at different population doubling levels, especially at replicative senescence, which limits the suitability of this method for the analysis of normal human cells. Since molecules responsible for autofluorescence are predominantly accumulating in the cytoplasm, we decided to isolate the nuclei to perform flow FISH analysis. With this novel nuclear flow FISH (NFF) technique we were able to minimize autofluorescence and its variability, thereby improving the signal-to-noise ratio and consequently, allowing the determination of telomere length during in vitro aging with high accuracy. Moreover, NFF will find broader applications, whenever in situ hybridization signals have to be quantitated. PMID- 16311001 TI - Evaluation of LC-MS for the analysis of cleaning verification samples. AB - The cleaning verification of pharmaceutical manufacturing equipment prior to further use is a cGMP requirement. Typically, relevant data are generated by HPLC with UV detection using methods individually developed and validated for each product. This work describes the use of HPLC with mass spectrometry to analyse cleaning verification samples, a novel means of utilising this analytical technology. The initial aim was to produce a single, generic method capable of quantifying a broad range of pharmaceuticals. Ultimately, however, a more effective strategy, in terms of efficiency and reliability, proved to be application of a well-defined approach to the rapid generation of compound specific methods. Results of studies to optimise the sample preparation for a basic compound in drug development (compound 1), together with experimental results for two further compounds are presented. These demonstrated that the combination of a well defined approach to chromatographic method development and mass spectrometric detection provided methodology with advantages in terms of sensitivity. Additionally, and by virtue of its potential for general applicability, the approach proposed has the potential to improve the overall efficiency with which methods for cleaning verification samples can be developed and applied. PMID- 16311002 TI - Development and validation of an HPLC method for the determination of gatifloxacin stability in human plasma. AB - A simple reversed-phase high performance liquid chromatography (HPLC) method for the determination of gatifloxacin stability in human plasma was developed and validated. Using ciprofloxacin as an internal standard (IS), separation was achieved on X Terra MS C18 (3 mm x 50 mm, 5 microm) column. The mobile phase, 0.025 M disodium hydrogen phosphate (pH 3.0) and acetonitrile (80:20 v/v), were delivered at a flow rate of 1.0 ml/min. The eluent was monitored using spectrophotometeric detection at 293 nm. Plasma samples were deproteinized using Amicon Centrifree system. No interference in blank plasma or of commonly used drugs was observed. The relationship between gatifloxacin concentration and peak height ratio of gatifloxacin to the IS was linear over the range of 0.10-6.0 microg/ml. The intra-day and inter-day coefficients of variation were < or = 2.77 and < or = 4.59%, respectively. The extraction recovery of gatifloxacin and the IS from plasma samples was > or = 85%. Gatifloxacin was found to be stable for at least 5 h at RT, 7 weeks at -20 degrees C, and after 3 freeze-thaw cycles in plasma; 16 h at RT and 48 h at -20 degrees C in deproteinized plasma; and 24 h at RT and 7 weeks at -20 degrees C in phosphate buffer. PMID- 16311003 TI - Enantioselective determination of a gastroprokinetic drug using amylose tris-(3,5 dimethylphenylcarbamate) as a stationary phase by HPLC. AB - An enantioselective high-performance liquid chromatographic method for determination of enantiomers of mosapride citrate in bulk drugs and pharmaceuticals using UV-vis and polarimetric detectors in series has been developed. Baseline separation with resolution >2.0 was achieved on a column containing amylose tris-(3,5-dimethylphenylcarbamate) as stationary phase using a mobile phase consisting of n-hexane:ethanol:triethylamine (80:20:0.3, v/v/v) at 40 degrees C. The detection was carried out at UV-276 nm and enantiomers were identified by polarimetric detector. The effect of ethanol, 2-propanol, TEA, temperature and mobile phase flow rate on separation of MSP enantiomers was studied and the method was validated with respect to accuracy, precision, linearity and limits of detection and quantification. The linearity of the method was studied between 6.25 and 50 microg/ml and r2 was >0.9997. The recoveries were in the range 99.63-100.22%, the method was suitable not only for process development of mosapride citrate but also for quality assurance of the individual enantiomers in bulk drugs and pharmaceuticals. PMID- 16311004 TI - Development and validation of an automated solid phase extraction and liquid chromatographic method for the determination of piperaquine in urine. AB - A sensitive and specific bioanalytical method for determination of piperaquine in urine by automated solid-phase extraction (SPE) and liquid chromatography (LC) has been developed and validated. Buffered urine samples (containing internal standard) were loaded onto mixed phase (cation-exchange and octylsilica) SPE columns using an ASPEC XL SPE robot. Chromatographic separation was achieved on a Chromolith Performance RP-18e (100 mm x 4.6 mm I.D.) LC column with phosphate buffer (pH 2.5; 0.1 mol/L)-acetonitrile (92:8, v/v). Piperaquine was analysed at a flow rate of 3 mL/min with UV detection at 347 nm. A linear regression model on log-log transformed data was used for quantification. Within-day precision for piperaquine was 1.3% at 5000 ng/mL and 6.6% at 50 ng/mL. Between-day precision for piperaquine was 3.7% at 5000 ng/mL and 7.2% at 50 ng/mL. Total-assay precision for piperaquine over 4 days using five replicates each day (n = 20) was 4.0%, 5.2% and 9.8% at 5000, 500 and 50 ng/mL, respectively. The lower limit of quantification (LLOQ) was set to 3 ng/mL using 1 mL of urine, which could be lowered to 0.33 ng/mL when using 9 mL of urine and an increased injection volume. PMID- 16311005 TI - [Haemorrhagic shock during cerebrospinal fluid shunt procedure. Reassessment of the anaesthetic or surgical practice?]. AB - We report two cases of perioperative haemorrhagic shock after accidental puncture of trunc vessels, during emergency shunt procedures in children who suffer of severe intracranial hypertension. In both cases it's the peritoneal internalization of the shunt with Portnoy trocar which is responsible of these deep vascular wounds. Evolution was favourable in both cases with cardiovascular resuscitation, transfusion and surgical haemostatic correction. Few days later, an internal shunt was performed in the second patient, while the first patient did not need shunt anymore. These two accidents lead to the discussion of the surgical procedure with surgeons, in order to have a better prevention against this complication. We also discuss our anaesthesiological practice from preoperative to perioperative period of this usually non-haemorrhagic surgical procedure. PMID- 16311006 TI - [Intrathecal use of colistin]. PMID- 16311007 TI - [Easy tracheal intubation? Mallampati class zero airway]. PMID- 16311008 TI - [Postpartum ovarian vein thrombophlebitis revealed by pulmonary signs]. PMID- 16311009 TI - Investigation into the ability of the Salmonella umu test to detect DNA damage using antitumor drugs. AB - In order to examine the ability of the umu test detecting system, 18 antitumor drugs were tested using the Salmonella umu test. The tested antitumor drugs were selected so as to produce different biochemical actions, and they were classified into three categories; five agents of group I (antimetabolites), eight agents of group II (alkylating agents), and five agents of group III (antibiotics). The results showed that all antimetabolites, all alkylating agents, and three of the antibiotics had positive responses, but the antibiotics aclarubicin (ACR) and chromomycin A3 (CHR) had negative responses. Both antibiotics that gave negative responses were anthracyclines, but daunomycin (DNR), which was one of the anthracyclines, had a positive result in the umu test. These results suggest that it is possible for the umu test to detect genotoxicity of chemicals regardless of the types of DNA damage (inhibition of DNA synthesis relative enzyme, DNA base alkylating, DNA strand-break, and DNA adduct), but difficult for it to detect genotoxicity of any anthracyclines. PMID- 16311010 TI - Correlation of apoptotic potential of simple oxiranes with cytotoxicity. AB - The measurement of primary DNA damage caused by oxirane chemicals can be confounded by apoptotic-generated DNA autolysis. The apoptogenic potential of oxiranes requires knowledge of the relationship between the apoptotic threshold dose and cytotoxic dose for interpretation of DNA damage assays. This research determined the relationship between cytotoxic and apoptotic doses for seven simple oxiranes of varying structure. This relationship between cytotoxic and apoptotic thresholds was determined simultaneously in in vitro cell culture. L929 cells in log-phase growth were exposed to the oxiranes for 24 h in 25 cm(2) and then assayed fluorometrically in 96-well plates for Caspase 3. Viability was assessed using Trypan Blue exclusion and loss of Caspase 3 activity. Ranked apoptotic potency was: diepoxybutane (DEB)>styrene oxide (SO)>phenyl glycidyl ether (PGE)>epichlorhydrin (EPI)>glycidol (GLY)>epoxybutane (EB)>epoxycyclohexane (ECH). Relative cytotoxicity was significantly correlated (r(s)=0.86, p=0.02) with potencies: DEB>EPI>PGE>SO>GLY>EB>ECH. These structurally-diverse, simple oxiranes were all capable of inducing apoptosis at doses several-fold below their cytotoxic concentrations. Difunctionality and aromaticity were key predictors of potency for both. Caspase 3 activity was an accurate indicator of necrosis which correlated with Trypan Blue results. PMID- 16311011 TI - Development of an in vitro skin sensitization test using human cell lines: the human Cell Line Activation Test (h-CLAT). I. Optimization of the h-CLAT protocol. AB - The aim of this study is to optimize the experimental conditions for an in vitro skin sensitization test using the human cell lines THP-1 and U-937. As regards pre-culturing time, the expression of CD86 on DNCB-treated THP-1 cells tended to be higher after 48h and 72h pre-culture compared with other time points evaluated. Next, we investigated the effect of chemical treatment time, and found that induction of CD86 expression on THP-1 cells by DNCB reached a plateau after 24h. Augmentation of CD86 expression is often observed when cells are treated with a subtoxic dose of allergens. To determine the appropriate dose of test samples, the cytotoxicity of test samples to THP-1 and U-937 cells was assessed with MTT assay, and the 50% inhibitory concentration (IC50) of each test sample was calculated. Based on the cytotoxicity assay data, four concentrations in the range between toxic and non-toxic were selected (0.1x, 0.5x, 1x and 2x IC50). Several kinds of antibodies were tested for staining THP-1 and U-937 cells treated with allergens/non-allergens (e.g., DNCB, Ni/SLS), and suitable antibodies for staining CD86 and CD54 were selected. We confirmed that the working dilutions of the selected CD86 and CD54 antibodies were appropriate for use in our method. The effect of an FcR blocking procedure was also evaluated. The mean fluorescence intensity (MFI value) was decreased by the FcR blocking procedure, which indicated that non-specific staining was blocked. Therefore, this procedure should be included in the method. Based on our findings, the protocol for this assay was optimized and the experimental conditions to be used in a future validation study were identified. We propose to call this kind of in vitro skin sensitization test h-CLAT, which is short for human Cell Line Activation Test. PMID- 16311012 TI - Evaluation of cytogenetic and DNA damage induced by the antibacterial drug, trimethoprim. AB - Trimethoprim, a widely used antimicrobial drug was tested for its effect on the level of nuclear DNA damage in cultured peripheral blood lymphocytes in terms of chromosome and DNA alterations. The extent of cytogenetic damage, expressed as chromosome breakage and chromosome loss, was evaluated employing the cytokinesis block micronucleus method (CBMN) in cultured peripheral blood lymphocytes coupled with fluorescence in situ hybridization (FISH) using a digoxigenin-labelled alphoid DNA probe specific for the centromere of all human chromosomes. The DNA breakage level was evaluated by the Comet assay. Cultures were set up by using blood samples from two healthy donors. A range of concentrations of the test agent (from 1 to 100 microg/ml) was used for the micronuclei (MN) frequency and cytogenetic origin of MN. For the Comet assay the range of doses used was from 0.5 to 150 microg/ml. From the results obtained it appears that this antifolic agent has a significant clastogenic potential, as detected by a dose-dependent increase of the incidence of C-MN and significantly greater than control levels at the highest concentrations tested (25,100 microg/ml). In addition, the results obtained in the Comet assay also show that trimethoprim induces a dose-dependent increase in the level of DNA breakage, this increase attaining statistical significance at the highest concentrations tested (25, 100, 150 microg/ml), which would confirm its genotoxicity. PMID- 16311013 TI - Association of Reelin gene polymorphisms with autism. AB - Genome scans indicate a linkage of autism to the chromosome 7q21-q36 region. Recent studies suggest that the Reelin gene may be one of the loci contributing to the positive linkage between chromosome 7q and autism. However, these studies were relatively small scale, using a few markers in the gene. We investigated 34 single nucleotide polymorphisms (SNPs) in the Reelin gene with an average spacing between the SNPs of 15 kb for evidence of association with autism. There were significant differences in the transmission of the alleles of exon 22 and intron 59 SNP to autistic subjects. Our findings support a role for the Reelin gene in the susceptibility to autism. PMID- 16311014 TI - Tag/anti-tag liquid-phase primer extension array: a flexible and versatile genotyping platform. AB - This study demonstrates an array-based platform to genotype simultaneously single nucleotide polymorphisms (SNPs) and some short insertions/deletions (indels) by the integration of the universal tag/anti-tag (TAT) system, liquid-phase primer extension (LIPEX), and a novel two-color detection strategy on an array format (TATLIPEXA). The TAT system permits a universal chip to be used for many applications, and the LIPEX simplifies the sample preparation but improves the sensitivity significantly. More importantly, all SNPs and some short indels can be interrogated in a single reaction with only two fluorescent ddNTPs. The concept of TATLIPEXA is demonstrated for nine SNPs (eight point mutations and one single-base insertion), and genotypes obtained show a remarkable concordance rate of 100% with both DNA sequencing and restriction fragment length polymorphism. Moreover, TATLIPEXA is able to provide quantitative information on allele frequency in pooled DNA samples, which could serve as a rapid screening tool for SNPs associated with diseases. PMID- 16311015 TI - Bioinformatic characterization of the SynCAM family of immunoglobulin-like domain containing adhesion molecules. AB - SynCAM 1 (synaptic cell adhesion molecule 1, alternatively named Tslc1 and nectin like protein 3) belongs to the immunoglobulin superfamily and is an adhesion molecule that operates in a variety of important contexts. Exemplary are its roles in adhesion at synapses in the central nervous system and as tumor suppressor. Here, I describe a family of genes homologous to SynCAM 1 comprising four genes found solely in vertebrates. All SynCAM genes encode proteins with three immunoglobulin-like domains of the V-set, C1-set, and I-set subclasses. Comparison of genomic with cDNA sequences provides their exon-intron structure. Alternative splicing generates isoforms of SynCAM proteins, and diverse SynCAM 1 and 2 isoforms are created in an extracellular region rich in predicted O glycosylation sites. Protein interaction motifs in the cytosolic sequence are highly conserved among all four SynCAM proteins, indicating their critical functional role. These findings aim to facilitate the understanding of SynCAM genes and provide the framework to examine the physiological functions of this family of vertebrate-specific adhesion molecules. PMID- 16311016 TI - Mouse cDNA microarray analysis uncovers Slug targets in mouse embryonic fibroblasts. AB - There is a need to reveal mechanisms that account for maintenance of the mesenchymal phenotype in normal development and cancer. Slug (approved gene symbol Snai2), a member of the Snail gene family of zinc-finger transcription factors, is believed to function in the maintenance of the nonepithelial phenotype. This study identified candidate Slug target genes linked to Slug gene suppression in primary mouse embryonic fibroblasts. Expression analyses were performed with a mouse cDNA microarray (Mousechip-CNIO) containing 15,000 clones. A total of 15 novel Slug target species were validated by real-time PCR or Western analyses. These included self-renewal genes (Bmi1, Nanog, Gfi1), epithelial-mesenchymal genes (Tcfe2a, Ctnb1, Sin3a, Hdac1, Hdac2, Muc1, Cldn11), survival genes (Bcl2, Bbc3), and cell cycle/damage genes (Cdkn1a, Rbl1, Mdm2). Expression patterns were studied in wild-type MEFs and Slug-deficient MEFs. Slug complementation studies recovered aberrant gene expression in cells lacking Slug, indicating that these genes were regulated directly by Slug. These results highlight their potential roles in mediating Slug function in mesenchymal cells and may help to identify novel therapeutic biomarkers in cancers linked to Slug. PMID- 16311017 TI - Congenital toxoplasmosis--prenatal aspects of Toxoplasma gondii infection. AB - Toxoplasma gondii (T. gondii) is the cause of toxoplasmosis. Primary infection in an immunocompetent person is usually asymptomatic. Serological surveys demonstrate that world-wide exposure to T. gondii is high (30% in US and 50-80% in Europe). Vertical transmission from a recently infected pregnant woman to her fetus may lead to congenital toxoplasmosis. The risk of such transmission increases as primary maternal infection occurs later in pregnancy. However, consequences for the fetus are more severe with transmission closer to conception. The timing of maternal primary infection is, therefore, critically linked to the clinical manifestations of the infection. Fetal infection may result in natural abortion. Often, no apparent symptoms are observed at birth and complications develop only later in life. The laboratory methods of assessing fetal risk of T. gondii infection are serology and direct tests. Screening programs for women at childbearing age or of the newborn, as well as education of the public regarding infection prevention, proved to be cost-effective and reduce the rate of infection. The impact of antiparasytic therapy on vertical transmission from mother to fetus is still controversial. However, specific therapy is recommended to be initiated as soon as infection is diagnosed. PMID- 16311018 TI - Effects of bisphenol A given neonatally on reproductive functions of male rats. AB - Male Sprague-Dawley rats (Crj:CD (IGS) were treated neonatally with bisphenol A (BPA) to evaluate effects on reproductive parameters. Animals were given BPA subcutaneously in corn oil to dosages of 0.002-97 mg/kg body weight, or 0.9 mg/kg 17beta-estradiol (E2) once a day from postnatal day (PND) 0 to PND 9. Preputial separation, copulatory rate, fertility rate, sperm analysis, serum testosterone levels, and gene expression in the testis were assessed. Males in the E2 group showed a decrease in testis weight and alterations of estrogen-mediated gene expression in the testis on PND 10, and by PND 150 incomplete preputial separation, decreases in the copulatory rate, testicular and accessory organ weights and number of sperm. In contrast, males in all BPA groups showed normal reproductive parameters. These results indicate that in male rats, BPA given during the neonatal period neither affected reproductive function nor evoked estrogen-mediated gene responses in the testis. PMID- 16311019 TI - Pharmacogenetics of autoimmune diseases: research issues in the case of Multiple Sclerosis and the role of IFN-beta. AB - Pharmacogenetics of auto-immune diseases is a complex field of application for this relatively new discipline, since we still have a partial knowledge of the biological mechanisms of the disease and of the drugs currently used to treat it. We address a few key issues that emerge when planning a pharmacogenetic investigation in Multiple Sclerosis and that relate to the complexities existing at the biological-genetic level and at the phenotypic characterization. In fact, we think that a clearer characterization of the clinical phenotype representing the end-point of the investigation together with a critical appraisal of the multi-faceted dimension of the genetic component of either the disease and the pharmacogenetic profile of the drug investigated, will help to design more thorough study and to achieve deeper understanding of the practical results. We will primarily focus our research considerations on the role of Interferon Beta (IFN-beta) as a prototypal therapeutic agent in Multiple Sclerosis. PMID- 16311020 TI - Drosophila TAB2 is required for the immune activation of JNK and NF-kappaB. AB - The TAK1 plays a pivotal role in the innate immune response of Drosophila by controlling the activation of JNK and NF-kappaB. Activation of TAK1 in mammals is mediated by two TAK1-binding proteins, TAB1 and TAB2, but the role of the TAB proteins in the immune response of Drosophila has not yet been established. Here, we report the identification of a TAB2-like protein in Drosophila called dTAB2. dTAB2 can interact with dTAK1, and stimulate the activation of the JNK and NF-kB signaling pathway. Furthermore, we have found that silencing of dTAB2 expression by dsRNAi inhibits JNK activation by peptidoglycans (PGN), but not by NaCl or sorbitol. In addition, suppression of dTAB2 blocked PGN-induced expression of antibacterial peptide genes, a function normally mediated by the activation of NF kappaB signaling pathway. No significant effect on p38 activation by dTAB2 was found. These results suggest that dTAB2 is specifically required for PGN-induced activation of JNK and NF-kappaB signaling pathways. PMID- 16311021 TI - Examination of the zinc transporter gene, SLC39A12. PMID- 16311022 TI - VSL#3 probiotic preparation has the capacity to hydrolyze gliadin polypeptides responsible for Celiac Sprue. AB - The native structure and distribution of gliadin epitopes responsible for Celiac Sprue (CS) may be influenced by cereal food processing. This work was aimed at showing the capacity of probiotic VSL#3 to decrease the toxicity of wheat flour during long-time fermentation. VSL#3 (10(9) cfu/ml) hydrolyzed completely the alpha2-gliadin-derived epitopes 62-75 and 33-mer (750 ppm). Two-dimensional electrophoresis, immunological (R5 antibody) and mass spectrometry analyses showed an almost complete degradation of gliadins during long-time fermentation of wheat flour by VSL#3. Gliadins non-hydrolyzed during fermentation by VSL#3 were subjected to peptic-tryptic (PT) digestion and analyzed by CapLC-ESI-Q-ToF MS (Capillary Liquid Chromatography-Electrospray Ionization-Quadrupole-Time of Flight-Mass Spectrometry). Search for several epitopes showed the only presence of alpha2-gliadin-fragment 62-75 at a very low concentration (sub-ppm range). Compared to IEC-6 cells exposed to intact gliadins extracted from the chemically acidified dough (control), VSL#3 pre-digested gliadins caused a less pronounced reorganization of the intracellular F-actin which was mirrored by an attenuated effect on intestinal mucosa permeability. The release of zonulin from intestinal epithelial cells treated with gliadins was considerably lower when digested with VSL#3. Agglutination test on K 562 (S) cells showed that the PT-digest of wheat flour treated with VSL#3 increased the Minimal Agglutinating Activity of ca. 100 times. Wheat proteins were extracted from doughs and subjected to PT digestion. Compared to PT-digest from chemically acidified dough, celiac jejunal biopsies exposed to the PT-digest from the dough fermented by VSL#3 did not show an increase of the infiltration of CD3(+) intraepithelial lymphocytes. Proteolytic activity by probiotic VSL#3 may have an importance during food processing to produce pre-digested and tolerated gliadins for increasing the palatability of gluten-free products. PMID- 16311023 TI - Female hyper IgM syndrome type 1 with a chromosomal translocation disrupting CD40LG. AB - Hyper-IgM syndrome type 1 (HIGM1) is a primary immunodeficiency characterized by recurrent bacterial and opportunistic infections, associated with normal or high serum level of IgM and decreased serum levels of IgG, IgA and IgE due to the defect of class switch recombination. CD40LG, located in Xq26, has been reported to be mutated in male HIGM1 patients. Here, we report the second case of a female HIGM1 with the defect of CD40 ligand (CD40L) expression and of soluble serum CD40L. Clinical course and HIGM phenotype was indistinguishable from that of male HIGM1 including severe neutropenia. High-resolution chromosome banding revealed that this patient's karyotype is 46, X, t(X;14)(q26.3;q13.1), and FISH analysis demonstrated that the break point of the chromosomal translocation is within CD40LG. Using four chimeric cDNA clones obtained by 3' RACE method, the break point was identified within the intron 4 of CD40LG on X chromosome and non-coding region of chromosome 14. We also found an extremely skewed X-chromosome inactivation pattern by methylation-specific PCR. Thus, the reciprocal translocation caused the disruption of CD40LG, resulting in defective CD40L expression in the female patient with an extremely skewed X-inactivation pattern in T cells leading to the HIGM1 phenotype. PMID- 16311024 TI - Influence of cellulose powder structure on moisture-induced degradation of acetylsalicylic acid. AB - The stability of crystalline acetylsalicylic acid (ASA) powder in binary mixtures with cellulose powders was investigated to reveal information about the influence of the cellulose structural properties on the moisture-induced ASA degradation. Different cellulose powder samples were manufactured and characterized by X-ray diffraction and N2 BET gas adsorption. The degradation patterns in ASA/cellulose mixtures were monitored as a function of salicylic acid increase versus time under various relative humidity conditions at 50 degrees C. The crystallinity index of cellulose samples varied between approximately 49 and 95%. The results indicated that cellulose powder with the lowest crystallinity index exhibited lower degradation rates than the samples with the higher crystallinity index. It should be noted that higher ASA degradation rates were observed in the samples with comparably lower moisture contents. This effect was most pronounced in the 1:3 (w/w), ASA/cellulose mixtures, whereas in 3:1 (w/w), ASA/cellulose mixtures the effect was less obvious. The findings emphasise the importance of cellulose structural organisation when governing the moisture's partition between cellulose and ASA during the hydrolytic degradation. PMID- 16311025 TI - [Pulmonary involvement with Crohn's disease in an adolescent]. PMID- 16311026 TI - The influence of underweight and dietary support on well-being in lung transplant candidates. AB - BACKGROUND: Quality of life has been found to be a significant predictor of survival in lung transplantation candidates. The aim of this study was to investigate associations between underweight, dietary support and well-being. METHODS: A self-administered questionnaire for perceived well-being was administered to underweight (n=42) and normal-weight (n=29) candidates for lung transplantation before and after dietary intervention in which the underweight patients received dietary support for weight gain. RESULTS: Underweight compared with normal-weight, independent of lung function, was associated with low well being in several of the measured dimensions. Improvements were observed after dietary intervention compared with baseline in the underweight patients, for scores in the dimension of tiredness 29.2 (4.2) vs. 26.2 (6.0), P<0.01; general satisfaction 4.7 (1.5) vs. 4.0 (1.4), P=0.01; social life 16.7 (3.9) vs. 15.0 (4.4), P=0.02) (mean (sd) before and after dietary intervention respectively), but not in the normal-weight patients. The underweight patients achieved the goal for energy intake and protein intake and experienced a significant weight gain. Regression analyses showed that none of the well-being improvements was associated with weight gain or change in body composition. However, an association between less tiredness and an increase in protein intake was indicated (b=-0.305, P=0.055). CONCLUSION: Underweight compared with normal weight was associated with more impaired quality of life in candidates for lung transplantation and some benefit from dietary support in terms of well-being was indicated. PMID- 16311027 TI - Genetic background influences metabolic response to dietary phosphorus restriction. AB - Dietary phosphorus (P) is essential to bone growth and turnover; however, little research has focused on the genetic mechanisms controlling P utilization. Understanding the interactions between genetics and dietary P that optimize bone integrity could provide novel interventions for osteoporosis. Thirty-six pigs from two sire lines known to differ in bone structure [heavier boned (HB) and lighter boned (LB)] were assigned to one of the three diets (P adequate, P repletion or P deficient). After 14 days, bone marrow and intact radial bones were collected. Differences between these lines in growth rate, bone integrity and gene expression within bone marrow were observed. In HB, but not LB, pigs, the P-deficient diet decreased weight gain (P<.01). For both lines, P deficiency caused a reduction in radial bone strength (P<.01), but HB P-deficient animals had greater (P<.10) bone integrity than P-deficient LB pigs. In HB, but not LB, pigs, dietary treatment affected the expression of CALCR (calcitonin receptor) (P<.05), VDR (vitamin D receptor) (P<.04) and IGFBP3 (insulin-like growth factor binding protein 3) (P<.06). There was also a trend of increased IL6 (interleukin 6), TFIIB (transcription initiation factor IIB) and SOX9 (sex determining region Y-box 9) expression with P deficiency in HB, but not LB, pigs. Both genetic backgrounds responded similarly to P deficiency with an increase in the expression of OXTR (oxytocin receptor) and IGF1 (insulin-like growth factor 1). Differences in growth rate, bone integrity and gene expression within the bone marrow suggest a difference in the homeorhetic control of P utilization between these genetic lines. Understanding these differences could lead to novel treatments for osteoporosis and aid in the development of tests for identifying those at risk for this disease. PMID- 16311028 TI - Seasonal variation in the chemical composition of two tropical seaweeds. AB - The chemical composition of red seaweed Gracilaria cervicornis and brown seaweed Sargassum vulgare from Brazil was investigated. In this study, the relationship between the nutritive components of each species and the environment was established. Protein content varied from 23.05+/-3.04% to 15.97+/-3.04%. The highest value was found in G. cervicornis. The protein levels were positively correlated with nitrogen content and negatively with water temperature and salinity. Carbohydrate contents of both species varied significantly (p<0.01) and the values observed were superior to others chemical constituents. Contrary to carbohydrates, the lipid concentrations were the lowest recorded chemical component and varied slightly between the two species. Ash content was greater in S. vulgare (14.20+/-3.86) than in G. cervicornis (7.74+/-1.15). In general the variation in chemical composition was related to environment. PMID- 16311029 TI - Development of sludge filterability test to assess the solids removal potential of a sludge bed. AB - A qualitative sludge characterisation technique called "sludge filterability technique" has been developed. This technique enables the determination of the sludge potential for the physical removal of solids, weighing the effect of different process parameters on solids removal and identifying the mechanisms of solids removal in an upflow anaerobic sludge bed system. In this paper guidelines for conducting the test are given and a "standardised" set-up is presented. The experimental set-up and protocol are simple and the results can be obtained in a period as short as a few hours. A sludge sample is added to an upflow reactor incubated at 4 degrees C, to limit gas production, washed with an anaerobically pre-treated and suspended solids free wastewater to remove solids washed out from the sludge, and then fed with a model substrate, prepared from fish meal with a standard procedure. Several experimental runs were conducted to validate and optimise the technique. The results showed that the technique is reliable, workable and reproducible. PMID- 16311030 TI - Relationship between density and ultrasonic velocity in Brazilian tropical woods. AB - In this study, the effect of density on the velocity of an ultrasonic wave in wood is investigated. The aim of the present study was to analyze the influence of density on the longitudinal velocity of an ultrasonic wave, propagated in the longitudinal direction. Experiments were conducted on 5cm x 5cm x 5cm wood specimens selected from the following species: pinus caribea (Pinus caribea var. caribea), eucalyptus citriodora (Eucalyptus citriodora), eucalyptus grandis (Eucalyptus grandis), cupiuba (Goupia glabra) and jatoba (Hymenaea sp.). The relationship between density and velocity was analyzed in two different manners: between and within species. The results obtained between species indicated that ultrasonic velocity tends to increase with increasing density. The results obtained within species also showed an increasing trend in ultrasonic velocity as density increased, but the relationship was not as significant as it was for between species. PMID- 16311031 TI - Potential utilization of guar gum industrial waste in vermicompost production. AB - Recycling of guar gum industrial waste through vermitechnology was studied under laboratory conditions by using composting earthworm Perionyx excavatus (Perrier). Three different combination of guar gum industrial waste namely guar gum industrial waste:cow dung:saw dust in 40:30:30 ratio (T1), guar gum industrial waste:cow dung:saw dust in 60:20:20 ratio (T2), and guar gum industrial waste:cow dung:saw dust in 75:15:10 ratio (T3) were used for vermicomposting experiments. Chemical changes during vermicomposting were measured and comparatively T2 showed great increase (from its initial level) for total N (25.4%), phosphorus (72.8%) and potassium (20.9%) than the other treatments. T2 also showed higher vermicomposting coefficient (VC), higher mean biomass for P. excavatus (146.68 mg) and higher cocoon production (about 21.9% and 645.5% more than the T1 and T3, respectively). Maximum earthworm mortality during vermicomposting was recorded with T3 treatment while zero mortality was recorded for T2 treatment after 150 days. Overall, T2 treatment appeared to be an ideal combination for enhancing maximum biopotential of earthworms to management guar gum industrial waste as well as for earthworm biomass and cocoon production. PMID- 16311032 TI - Cellulose acetates from linters and sisal: correlation between synthesis conditions in DMAc/LiCl and product properties. AB - We report the acetylation of celluloses from sisal (untreated and alkali treated) and cotton linters (alkali treated), under homogeneous solution conditions, using DMAc/LiCl as solvent system. Our target was to evaluate the effects of cellulose dissolution and reactions conditions on the product properties. The products were characterized in terms of degree of substitution (DS) by 1H NMR, and molar weight distribution (MWD) by size exclusion chromatography. Changes in the DS of the products were correlated with reaction conditions and solution properties. It was found that the dissolution of celluloses and degree of substitution of cellulose derivatives depends on a fine adjustment of the dissolution/derivatization conditions, as well as on the origin (sisal or linters) of celluloses. PMID- 16311033 TI - Thermodynamic and breakthrough column studies for the selective sorption of chromium from industrial effluent on activated eucalyptus bark. AB - Studies were carried out on adsorption of Cr(VI) on an adsorbent made from eucalyptus bark. Results revealed that sorption of chromium on activated eucalyptus bark (AEB) was endothermic in nature. Thermodynamic parameters such as the entropy change, enthalpy change and Gibb's free energy change were found out to be 100.97 J mol(-1)K(-1), 33 kJ mol(-1) and -0.737 kJ mol(-1), respectively. Industrial chrome effluent of different chromium concentration at different pH was used as feedstock for the fixed bed adsorption studies. When effluent was fed to the column at low pH of 2, the breakthrough volume increased significantly compared to effluent at higher pH of 4.85. The surface properties of sorbent were characterized by the Scanning electron microscopy, X-ray diffraction technique and Infrared techniques. It was concluded that AEB sorbent column could be used effectively for removal of chromium from industrial effluents by reducing the pH of chrome effluent to two and at optimal column conditions. PMID- 16311034 TI - Lobular carcinoma of the breast metastatic to the uterus in a patient under adjuvant anastrozole therapy. AB - This is the first report of breast carcinoma metastatic to the endometrium in a patient on adjuvant anastrozole therapy. We report a case of metastatic lobular carcinoma of the breast in a 63-year-old patient on adjuvant anastrozole therapy for 8 months. She was asymptomatic and metastatic endometrium was diagnosed after transvaginal ultrasound revealed suspicious findings along with elevated Ca 15-3 levels. As further work up showed no other metastatic sites her uterus was taken out along with her ovaries and pelvic lymph nodes. Uterine metastases should be kept in mind in asymptomatic patients on anastrozole therapy. PMID- 16311035 TI - Prediction methods to account for the effect of gait speed on lower limb angular kinematics. AB - Altering gait speed appears to have an effect on lower limb kinematics during gait. This is a potential source of inaccuracy when data recorded from healthy volunteers are compared with those of slow-walking patients. The aim of this study was to assess the accuracy of several prediction methods which could be used to predict appropriate kinematic data obtained at various walking speeds. Seventeen healthy adults were video-recorded while walking at three different speeds (slow, normal and fast). Five lower limb angles (knee, ankle, thigh, shank and foot) were assessed at 26 points in the gait cycle. Significant correlations were observed between angles and gait speed. The relationship between gait speed and angles was also dependent on the point in the gait cycle that was used for comparison. Results indicate that using prediction methods based on group mean data or regression data from the same speed range on which the prediction methods are being applied is significantly more accurate than prediction methods based on other speed ranges. These results indicate that 'ideal' angle data for slow walking speeds should be based on data recorded from slow-walking healthy adults or regressed using suitable regression equations. PMID- 16311036 TI - Changes in coordination of postural control during dynamic stance in chronic low back pain patients. AB - The human postural system operates on the basis of integrated information from three independent sources: vestibular, visual and somatosensory. It is conceivable that a derangement of any of these systems will influence the overall output of the postural system. The peripheral proprioceptive system or the central processing of proprioceptive information may be altered in chronic low back pain (CLBP). We therefore investigated whether patients with CLBP exhibited an altered postural control during quiet standing. Dynamic posturography was performed by 12 CLBP patients and 12 age-matched controls. Subject's task was to stand quietly on a computer-controlled movable platform under six sensory conditions that altered the available visual and proprioceptive information. While the control of balance was comparable between the two groups across stabilized support surface conditions (1-3), CLBP patients oscillated much more than controls in the anterior-posterior (AP) direction in platform sway referenced conditions (4-6). Control experiments ruled out that increased sway was due to pain interference. In CLBP patients, postural stability under challenging conditions is maintained by an increased sway in AP direction. This change in postural strategy may underlie a dysfunction of the peripheral proprioceptive system or the central integration of proprioceptive information. PMID- 16311037 TI - Internal-sensing machinery directs the activity of the regulatory network in Escherichia coli. AB - Individual cells need to discern and synchronize transcriptional responses according to variations in external and internal conditions. Metabolites and chemical compounds are sensed by transcription factors (TFs), which direct the corresponding specific transcriptional responses. We propose a classification of the currently known TFs of Escherichia coli based on whether they respond to metabolites incorporated from the exterior, to internally produced compounds, or to both. When analyzing the mutual interactions of TFs, the dominant role of internal signal sensing becomes apparent, greatly due to the role of global regulators of transcription. This work encompasses metabolite-TF interactions, bridging the gap between the metabolic and regulatory networks, thus advancing towards an integrated network model for the understanding of cellular behavior. PMID- 16311038 TI - A simple method for the absolute determination of uranium enrichment by high resolution gamma spectrometry. AB - A simple method for the determination of uranium enrichment using high-resolution gamma spectrometry is presented in this paper. The method relies solely on the gamma-ray emission probabilities of 235U and 234mPa, and an iterative procedure for the least squares fit of a polynomial to a set of experimentally determined data. To ensure the reliability of the 234mPa gamma-ray emission probabilities employed, a new determination of these probabilities was carried out using a combination of gamma spectrometry and Cerenkov counting of a purified 234Th solution. Using these new data, a maximum difference of approximately 5% has been found between the experimental and declared uranium enrichment in a set of solid and liquid samples containing uranium compounds. PMID- 16311039 TI - Multi-day radon signals with a radioactive decay limb -- occurrence and geophysical significance. AB - Multi-day signals, generally with duration of 2-10 days, are a prominent temporal variation type of radon (Rn) in geogas in the unsaturated zone. Rare multi-day Rn signals have been found which are characterized by: (a) a declining limb lasting up to 10 days which conforms to the radioactive decay of Rn, (b) recurs at the same location and (c) is recorded in diverse situations-volcanic and seismogenic. It suggested that a Rn blob is injected at a lower level on a steady upward flow of geogas whereby the rise and final fall of the signal are attributed to the edges of the blob while the central Rn-decay segment records the passing of the decaying blob itself. Rn-decay signals are a small subset of multi-day Rn signals which are considered as highly irregular and unusable for the understanding of geophysical processes. In difference, it is concluded that multi-day Rn signals are probably proxies of subtle geodynamic processes at upper crustal levels and are therefore significant for studying such processes. PMID- 16311040 TI - Evolution, development and timing of puberty. AB - The age of menarche has fallen as child health has improved. Although there is ample evidence of delayed puberty being associated with poorer childhood nutrition, menarche is also influenced by prenatal factors. In particular, early onset of puberty is reported in children who have migrated from developing to developed countries. Evolutionary perspectives suggest that these effects can be explained by adaptive mechanisms. They also provide an explanation for the human pubertal growth spurt. In the past few decades, as puberty has advanced, biological maturation has come to precede psychosocial maturation significantly for the first time in our evolutionary history Although this developmental mismatch has considerable societal implications, care has to be taken not to medicalize contemporary early puberty inappropriately. PMID- 16311041 TI - Staging and treatment of differentiated thyroid carcinoma with radiolabeled somatostatin analogs. AB - In patients with progressive metastatic (or recurrent) differentiated thyroid carcinoma that either do not take up radioiodine or are unresponsive to continued radioiodine therapy, staging is difficult and treatment options are few. However, in most of these patients uptake of radiolabeled somatostatin analogs is evident on somatostatin-receptor scintigraphy (SRS). Using SRS, patients with sufficient uptake of radiolabeled somatostatin analogs can be selected for high-dose peptide receptor radionuclide therapy (PRRT) as an alternative targeted-treatment option. PRRT with the beta-particle-emitting radionuclides (90)yttrium ((90)Y) and (177)lutetium ((177)Lu) gives the best results in terms of objective tumor response. Promising, novel, radiolabeled somatostatin analogs that have a broader receptor affinity profile and, thus, a potentially wider therapeutic range are being tested clinically. PMID- 16311042 TI - Firing properties of motor units during fatigue in subjects after stroke. AB - The purpose of this work was to investigate the electromyographic (EMG) fatigue representations in muscles of subjects after stroke at the level of motor unit, based on the analysis of mean power frequency (MPF) in the power density spectrum (PDS) for intramuscular EMG and our previous modeling and experiment studies on the neuromuscular transmission failure (NTF). NTF due to the local muscular fatigue had been captured in motor unit signals from healthy subjects during a submaximal fatigue contraction previously. In this study, the EMG signals for the biceps brachii muscles were collected by needle electrodes from the affected and unaffected arms of six hemiplegic subjects after stroke, and from the dominated arm of six healthy subjects during a full maximum voluntary contraction (MVC) and a subsequent 20% MVC. The MPF of EMG trials detected intramuscularly during the full and 20% MVCs, and the parameters of motor unit action potential trains (MUAPTs) during 20% MVC were analyzed in three groups: the normal (from healthy subjects), unaffected (from subjects after stroke), and affected (from subjects after stroke). It was found that during the full MVC the MPFs of the normal and unaffected groups decreased more than the affected when monitored by a moving time window of 2 s. The comparison on the overall MPF during the full MVC for these three groups over the whole time course of the EMG signal (18 s) were: the affected overall MPF was higher than the unaffected (P < 0.05); and the unaffected overall MPF was larger than the normal (P < 0.05). However, no significant decrease in MPF was found for these three groups during 20% MVC. The NTF was captured in most MUAPTs in the groups of the normal and unaffected rather than in the affected group, symbolized by the lowered rates of change (RCs) of firing rate (FR) (P < 0.05), more MUAPTs with positive RCs of maximum oscillation (MO) in MUAPT power density spectra (P < 0.05), and the significant higher RCs of minimum inter-pulse interval (MINI) (P < 0.05) in the normal and unaffected compared to the affected group. Enhanced neural drives to the motor units of the unaffected and affected groups were observed during 20% MVC, which possibly came from the bilateral neural inputs due to the disinhibition of the ipsilateral projections in subjects after stroke. For identifying the fatigue associated with NTF, the motor unit firing parameters, FR, MINI, and MO, were more sensitive than the MPF. The results obtained in this work provided a further understanding on the EMG of the fatigue processes in paretic and non-paretic muscles during voluntary contractions. PMID- 16311043 TI - Modulation of pre-programmed muscle activation and stretch reflex to changes of contact surface and visual input during movement to absorb impact. AB - The purpose of this study was to examine the extent of modification of the preactivation and stretch reflex response in ankle joint muscles to different contact surfaces and visual input during movement to absorb impact. Experimental movements like landing were performed using a special sliding apparatus. Seven subjects made landings on the hard surface (Hard-S) of a metal force platform or soft surface (Soft-S) of a foam cushion with eyes open or closed. The electromyographic activities from the medial gastrocnemius (MG), soleus (Sol), and tibialis anterior (TA) muscles, contact force, and ankle joint angle were recorded. The preactivation levels of MG and TA to Hard-S increased compared to Soft-S. After foot contact, dorsiflexion velocity, impulse, and responses of the stretch reflex in MG and Sol were significantly larger on Hard-S than Soft-S. With eyes closed, there were trends of decrease in the preactivation. Although the dorsiflexion velocity and impulse showed no significant differences between both visual conditions, the stretch reflex responses with eyes closed were larger than those with eyes open for both surfaces. These results suggest that the preactivation is modulated to different surface and the reflex gain is enlarged by visual suppression. PMID- 16311044 TI - Postural muscle activity patterns during standing at rest and on an oscillating floor. AB - Postural muscle activity pattern was examined in the eyes-closed state after adequate adaptation to floor anteroposterior oscillation. Twenty-three subjects were grouped almost evenly according to dominance of anterior or posterior postural muscles in the trunk and thigh during quiet stance. In the posterior dominant group, this dominance was maintained at every frequency in most subjects. In the anterior-dominant group, this dominance was maintained in most subjects at 0.1 and 0.5 Hz but changed to posterior dominance at 1.0 and 1.5 Hz in about half the subjects. Periodicity of muscle activity was evaluated by EMG amplitude spectrum at the floor oscillation frequency. Periodicity of posterior dominant muscles in the trunk and thigh increased with increasing oscillatory frequency. In the trunk, the periodicity did not differ significantly between posterior-dominant and anterior-dominant groups. However, in the thigh, periodicity was significantly lower in the anterior-dominant muscles. This was considered to be caused by nonperiodic alternating action of the anterior and posterior muscles. In the lower leg, posterior dominance was observed in quiet stance and at all oscillation frequencies. Periodicity of soleus and gastrocnemius increased at higher frequencies and was higher in gastrocnemius than in soleus. The periodicity difference between both muscles decreased with increasing oscillation frequency. PMID- 16311045 TI - Unconscious manipulation of free choice in humans. AB - Previous research has shown that subliminally presented stimuli accelerate or delay responses afforded by supraliminally presented stimuli. Our experiments extend these findings by showing that unconscious stimuli even affect free choices between responses. Thus, actions that are phenomenally experienced as freely chosen are influenced without the actor becoming aware of the manipulation. However, the unconscious influence is limited to a response bias, as participants chose the primed response only in up to 60% of the trials. LRP data in free choice trials indicate that the prime was not ineffective in trials in which participants chose the non-primed response as then it delayed performance of the incongruently primed response. PMID- 16311046 TI - Phylogeny of the Gadidae (sensu Svetovidov, 1948) based on their morphology and two mitochondrial genes. AB - Although Codfishes are probably one of the most studied groups of all teleost fishes worldwide owing to their great importance to fisheries, their phylogeny and classification are still far from being firmly established. In this study, we present phylogenetic relationships of 19 out of 22 genera traditionally included in the Gadidae based on the analysis of entire cytochrome b and partial cytochrome oxidase I genes (1530 bp). Maximum Parsimony, Maximum Likelihood, and Bayesian analyses all recovered five main clades that correspond to traditionally recognized groupings within Gadoids. The same clades were recovered with MP analysis based on 30 morphological characters (collected from the literature). Given these findings, we propose a revised provisional classification of Gadoids: one suborder Gadoidei containing two families, the Merlucciidae (1 genus) and the Gadidae (21 genera) distributed into four subfamilies: the Gadinae (12 genera), the Lotinae (3 genera), the Gaidropsarinae (3 genera), and the Phycinae (3 genera). Lastly, nuclear inserts of mitochondrial DNA (Numts) were identified in two species, i.e., Gadiculus argenteus and Melanogrammus aeglefinus. PMID- 16311047 TI - Novel in vitro perfusion system for the determination of hypothalamic-pituitary adrenal axis responses. AB - INTRODUCTION: The hypothalamic-pituitary-adrenal (HPA) axis is a three-gland component of the endocrine system and a key modulator of the stress response. We have developed a novel in vitro perfusion system to enable the study of pharmacological and hormonal challenges to tissue components of the HPA axis. In vivo studies have shown functional sex differences (sexual diergism) in HPA responses to cholinergic drugs, and in the present in vitro study, we examine these differences at several levels of the HPA axis. METHODS: Hypothalami, pituitaries, and adrenal glands were collected from male and female rats (n=3 per sex). One-half hypothalamus, one-half pituitary, and one adrenal gland were placed individually into three Erlenmeyer flasks connected by tubing. Flasks were perfused with medium (pH 7.4) at 37 degrees C. Sampling ports between the flasks were used to collect buffer for determination of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) release from the hypothalamus, pituitary, and adrenal flasks, respectively, over an extended baseline period, to determine stability of the system, and after nicotine administration. RESULTS: The perfusion system produced steady CRH, ACTH, and CORT baselines, the ACTH and CORT values being comparable to in vivo basal ACTH and CORT values in jugular-vein-cannulated rats. In vitro CRH, ACTH, and CORT responses to nicotine were significantly increased at 10 min and returned to baseline by 30 min, the CRH and ACTH responses from female tissues being greater than responses from male tissues. These sex differences were similar to those following nicotine administration in vivo. DISCUSSION: The ability of this novel, dynamic in vitro system to replicate in vivo HPA axis responses supports its potential as a new method for pharmacological and toxicological studies. PMID- 16311048 TI - Interleukin-17, a regulator of angiogenic factor release by synovial fibroblasts. AB - OBJECTIVE: Angiogenesis is a process stimulated in inflamed synovium of patients with osteoarthritis (OA), and contributes to the progression of the disease. Synovial fibroblasts secrete angiogenic factors, such as vascular endothelial growth factor (VEGF), an up-regulator of angiogenesis, and this ability is increased by interleukin (IL)-1beta. The purpose of this study was to verify whether IL-17 contributes and/or synergizes with IL-1beta and tumor necrosis factor (TNF)-alpha in vessel development in articular tissues by stimulating the secretion of proangiogenic factors by synovial fibroblasts. DESIGN: We stimulated in vitro synovial fibroblasts isolated from OA, rheumatoid arthritis (RA) and fractured patients (FP) with IL-17 and IL-1beta and from OA patients with IL-17, IL-1beta and TNF-alpha. In the supernatants from the cultures, we assayed the amount of VEGF by immunoassay and other angiogenic factors (keratinocyte growth factor, KGF; hepatocyte growth factor, HGF; heparin-binding endothelial growth factor, HB-EGF; angiopoietin-2, Ang-2; platelet-derived growth factor B, PDGF-BB; thrombopoietin, TPO) by chemiluminescence; semiquantitative RT-PCR was used to state mRNA expression of nonreleased angiogenic factors (Ang-2 and PDGF-BB) and tissue inhibitors of metalloproteinase (TIMP)-1. RESULTS: IL-17, TNF-alpha and IL 1beta increased VEGF secretion by synovial fibroblasts from OA patients. IL-17 and IL-1beta also increased VEGF secretion in RA and FP. Besides, IL-17 increased KGF and HGF secretions in OA, RA and FP; in OA and RA, IL-17 also increased the HB-EGF secretion and the expression of TIMP-1 as protein and mRNA. In OA patients IL-17 had an additive effect on TNF-alpha-stimulated VEGF secretion. CONCLUSIONS: These results suggest that IL-17 is an in vitro stimulator of angiogenic factor release, both by its own action and by cooperating with TNF-alpha. PMID- 16311049 TI - Differences in the gestational pattern of mRNA expression of the Rnd family in rat and human myometria. AB - Uterine myometrial contractility remains a poorly characterized area of research in reproductive physiology. Rnd1, a novel member of the GTP-binding Rho protein family, inhibits Ca(2+)-sensitization by specifically interfering with a RhoA/Rho activated kinases-dependent mechanism in smooth muscle. In addition to Rnd1, there are two other members, Rnd2 and Rnd3, in the Rnd family of Rho proteins. In the present comparative study of myometrial contractility in rats and humans, we found that all three Rnd mRNAs were expressed in nonpregnant rat myometrium and in nonpregnant human myometrial tissues. Although all three mRNA levels increased significantly after gestation in rat myometria, only Rnd1 expression was significantly greater after gestation in human samples. In the ovariectomized rat, administration of estrogen and/or progesterone increased the expression of all Rnd mRNAs. These results suggest that universal Rnd family up-regulation during pregnancy in rats may have an important role for negative-feedback control of uterine contraction during gestation by inhibiting RhoA-mediated increase in Ca(2+) sensitivity of contractile elements. Such increases in Rnd levels may be due to augmented levels of reproductive steroids in rats. Our data also point to gestational differences between rats and humans in Rnd isoform patterns. PMID- 16311050 TI - Two glutathione S-transferase isoenzymes purified from Bulinus truncatus (Gastropoda: Planorbidae). AB - We purified and characterized two major glutathione S-transferase isoenzymes (GST2 and GST3) from snail Bulinus truncatus (Mollusca, Gastropoda, Planorbidae) tissue. The Km with respect to 1-chloro-2, 4-dinitrobenzene (CDNB) for both isoenzymes was increased as the pH decreased. Km of both isoenzymes with respect to glutathione (GSH) doubled when the pH was increased from 6.0 to 6.5. Acid inactivated GST2 and GST3 and the two enzymes were almost inactive at pH 3.5. However, they retain the full activity for at least 20 h when incubated at pH between 6.0 and 9.0. The optimum temperature was 45 degrees C for GST2 and 50 degrees C for GST3. The half lifetime at 50 degrees C was 70 min and 45 min for GST2 and GST3 isoenzymes, respectively. Addition of 5 mM GSH to the incubation buffer increased the half life of both isoenzymes more than fourfold. The activation energy for catalyzing the conjugation of CDNB was 1.826 and 3.435 kcal/mol for GST2 and GST3, respectively. I50 values for Cibacron blue, bromosulphophthalein, indocyanine green, hematin and ethacrynic acid were 0.76 microM, 47.9 microM, 7.59 microM, 0.03 microM and 0.79 microM for GST2, and 0.479 microM, 79.4 microM, 89.1 microM, 32.4 microM and 1.15 microM for GST3, respectively. Cibacron blue and indocyanine green were non-competitive inhibitors, while hematin was a mixed inhibitor. Bromosulphophthalein was found to be a competitive inhibitor for GST2 and a mixed inhibitor for GST3. PMID- 16311051 TI - The cloned avian neurohypophysial hormone receptors. AB - Arginine vasotocin (AVT), a neurohypophysial hormone, has many essential functions in birds including the regulation of salt and fluid balance, blood pressure, the stress response and a variety of behaviors. In addition, AVT controls reproductive functions in birds that are served by oxytocin in mammals. In the following review, we examine the functions of AVT in birds with an emphasis on the present state of knowledge concerning the cloned receptors for this important hormone. Receptor and gene structure, signal transduction mechanisms and expression pattern are all discussed. Finally, we explore the phylogenetic relationships between the cloned avian receptors and other vertebrate and invertebrate neurohypophysial hormone receptors. PMID- 16311052 TI - Actin-inhibition and folding of vertebrate deoxyribonuclease I are affected by mutations at residues 67 and 114. AB - Amino acid (aa) residues (Val-67 and Ala-114) have been suggested as being mainly responsible for actin-binding in human and bovine deoxyribonucleases I (DNase I). This study presents evidence of these two aa mutational mechanisms, not only for actin-binding but also for folding of DNase I in mammals, reptiles and amphibians. Human and viper snake (Agkistrodon blomhoffii) enzymes are inhibited by actin, whereas porcine, rat snake (Elaphe quadrivirgata), and African clawed frog (Xenopus laevis) enzymes are not. To investigate the role of aa at 67, mutants of rat snake (Ile67Val) and viper snake (Val67Ile) enzymes were constructed. After substitution, the rat snake was inhibited by actin, while the viper snake was not. For the role of aa at 114, mutants of viper snake (Phe114Ala), rat snake (Phe114Ala), African clawed frog (Phe114Ala), and porcine (Ser114Ala/Ser114Phe) enzymes were constructed. Strikingly, the substitute mutants for viper snake, rat snake and African clawed frog expressed no protein. The porcine (Ser114Ala) enzyme was inhibited by actin, but not the porcine (Ser114Phe) enzyme. These results suggest that Val-67 may be essential for actin binding, that Phe-114 may be related to the folding of DNase I in reptiles and amphibians, and that Ala-114 may be indispensable for actin-binding in mammals. PMID- 16311053 TI - Regulation of insulin-like growth factor binding protein-5, four and a half lim 2, and a disintegrin and metalloprotease-9 expression in osteoblasts. AB - The roles of insulin-like growth factors (IGFs) in regulating growth and their modulation by six IGF binding proteins (IGFBP) are well established. IGFBP-5, the most abundant IGFBP stored in bone, is an important regulator of bone formation via IGF-dependent and -independent mechanisms. Two new proteins, four and a half lim (FHL)-2, a transcription modulator that interacts with IGFBP-5, and a disintegrin and metalloprotease (ADAM)-9, an IGFBP-5 protease, have been identified as potential regulators of IGFBP-5 action in bone. We tested the hypothesis that agents which modulate bone formation by regulating IGFBP-5 expression would also regulate FHL-2 and ADAM-9 expression in a coordinated manner. We evaluated the expression of IGFBP-5, FHL-2, and ADAM-9 by real-time reverse transcriptase (RT)-PCR during differentiation of mouse bone marrow stromal cells into osteoblasts and in response to treatment with bone formation modulators in the LSaOS human osteosarcoma cell line. IGFBP-5 and FHL-2 increased 4.3- and 3.0-fold (P < or = 0.01), respectively, during osteoblast differentiation. Dexamethasone (Dex), an inhibitor of bone formation, decreased IGFBP-5 and FHL-2 and increased ADAM-9 in LSaOS cells (P < or = 0.05). Bone morphogenic protein (BMP)-7, a stimulator of bone formation, increased IGFBP-5 and decreased ADAM-9 (P<0.01). To determine if BMP-7 would eliminate Dex inhibition of IGFBP-5, cells were treated with Dex+BMP-7. The BMP-7-induced increase in IGFBP-5 was reduced, but not eliminated, in the presence of Dex (P < or = 0.01), indicating that BMP-7 and Dex may regulate IGFBP-5 via different mechanisms. Transforming growth factor (TGF)-beta, a stimulator of bone formation, increased IGFBP-5 and FHL-2 expression (P < or = 0.01). IGF-I and TNF alpha decreased expression of ADAM-9 (P<0.05). In conclusion, our findings are consistent with the hypothesis that FHL-2 and ADAM-9 are important modulators of IGFBP-5 actions and are, in part, regulated in a coordinated manner in bone. PMID- 16311054 TI - Genetic/metabolic health care delivery during and after hurricanes Katrina and Rita. AB - Provision of health care to patients during and after events like those which occurred in association with hurricanes Katrina and Rita poses particular difficulties for rare disease patients, including those with genetic/metabolic diseases. In this summary, we recount the obstacles encountered in attempting to maintain and restore essential medical care to these patients, and offer proposals which may mitigate future such events. PMID- 16311055 TI - Helicobacter felis and Helicobacter bizzozeronii induce gastric parietal cell loss in Mongolian gerbils. AB - Non-pylori helicobacter infections are associated with gastritis, gastric ulcers and MALT lymphomas in man. Approximately 50% of these are caused by helicobacters commonly found in dogs and cats, including Helicobacter felis, Helicobacter bizzozeronii and H. salomonis. In contrast to Helicobacter pylori, the virulence mechanisms of these species are unknown. In this study the virulence of H. felis, H. bizzozeronii and H. salomonis was investigated in Mongolian gerbils. Female SPF gerbils were inoculated intragastrically with H. felis, H. bizzozeronii or H. salomonis and sacrificed 3 weeks later. Fundus and antrum samples were taken for bacterial detection by PCR. A longitudinal strip covering all stomach regions was taken for histology. Gastric colonization, inflammation, apoptosis, loss of parietal cells and cell proliferation were assessed. Controls and H. salomonis inoculated gerbils were negative in PCR. H. felis and H. bizzozeronii inoculated animals were positive. H. felis inoculated animals showed loss of parietal cells extending from the limiting ridge into the fundus. A high cell proliferation rate was noticed in the mucosal area devoid of parietal cells. A dense band of apoptotic cells and large numbers of Helicobacter bacteria were seen at the transition zone between affected and normal parietal cells. In H. bizzozeronii infected gerbils, this was less pronounced. Focal apoptotic loss of gastric epithelial cells was spatially associated with the presence of bacteria especially in H. felis and to a lesser extent in H. bizzozeronii infected gerbils. This loss of cells may lead to intestinal metaplasia. PMID- 16311056 TI - Bone metabolism and energy balance: role for leptin. PMID- 16311057 TI - Leptin, central nervous system, and bone: influence of physical activity. PMID- 16311058 TI - Central control of bone mass: potential therapeutic implications. PMID- 16311059 TI - The RATIO observatory: French registry of opportunistic infections, severe bacterial infections, and lymphomas complicating anti-TnFalpha therapy. AB - The RATIO observatory collects nationwide data on opportunistic infections, severe bacterial infections, and lymphomas in patients with a past or present history of tumor necrosis factor alpha (TNFalpha) antagonist treatment in France. The cases are validated by a committee of experts, and the capture-recapture method is used to check and to improve case ascertainment. A nested case-control comparison is carried out to identify risk factors for the events of interest. The registry differs from other biological registries in that the inclusion criterion is occurrence of the event (infection or lymphoma) instead of administration of the treatment. This method ensures collection of a far larger number of cases. The RATIO observatory is a remarkable example of a three-way partnership of learned societies, pharmaceutical companies, and institutions (the French research institute INSERM and the French drug safety agency AFSSAPS). Over 100 events were reported in the first 16 months, a large increase compared to European registries of fixed patient cohorts monitored for 4-5 years. This result validates our original approach, which will probably need to be extended to other biotherapies for inflammatory joint disease and to other potential adverse events. The strong commitment of rheumatologists in France, who are the main prescribers of TNFalpha antagonists, and of the French Society for Rheumatology explain the high case-ascertainment and must continue to ensure that answers are rapidly provided to the drug safety questions that are vital to our patients. PMID- 16311060 TI - [Indications of GnRH analogues before and after surgery for endometriosis]. AB - Indications of GnRH analogues before and/or after surgery for endometriosis remain controversial. Although some studies have underlined a decrease in rAFS scores using GnRH analogues pre-operatively, data are insufficient to recommend their systematic use in routine. In the same way, despite an increase in symptom free period subsequent to the use of GnRH analogues post-operatively, no effect of this therapeutic on recurrence rate and fertility outcome has been proved. In addition to a meta-analysis on GnRH analogues in endometriotic women, further studies, especially in women with large endometriomas and with deep pelvic endometriosis, are required to clarify the indications of GnRH analogues associated with surgical management. PMID- 16311061 TI - Investigation of thoracolumbar T12-L1 burst fracture mechanism using finite element method. AB - A finite element model of the T12-L1 motion segment was subjected to dynamic vertical impact to investigate vertebral burst fracture mechanism at the thoracolumbar junction. A rigid ball was directed vertically towards a rigid plate fixed on top of the T12 vertebral body to simulate the axial impact. The results show that upon impact, the T12 vertebra exhibited a vibratory motion. At its maximum compression, the endplates bulged towards their vertebral bodies. The central parts of the endplates adjacent to the nucleus experienced the highest effective stress, and localized stress concentration developed correspondingly within the central parts of the cancellous bone adjacent to the endplates. This appears to confirm the hypothesis that nucleus material is forced to enter the vertebral body, pressurizing it further and squeezing the fat and marrow contents out of the cancellous bone. When the nucleus material enters the vertebral body faster than fat and marrow being expulsed, the vertebral body could burst through the anterior and posterior cortical shell. Upon sudden posterior cortex fracture, the transient fragment encroachment could be further into the spinal canal than the final observed locations, as the fragments are retropulsed to the vertebral body during the bursting process. PMID- 16311062 TI - A fatal case of post-operative pulmonary thromboembolism with cosmetic liposuction. AB - Pulmonary thromboembolism (PTE) has been regarded as rare in Japan. However, PTE has been increasingly recognised because either of increased incidence, diagnostic progress, or social recognition. Recently, 10 Japanese Medical Associations have submitted preventive guidelines for PTE in post-operative patients and the government decided to fund this, as results of the increase in cases and concern regarding medical negligence. A fatal case of PTE after liposuction is reported. A female patient was in the home toilet after two days of immobilization following day-surgery liposuction. Clinicians must be aware of appropriate methods for the prevention of post operative PTE with cosmetic surgery. PMID- 16311063 TI - A qualitative and quantitative survey of Forensic Medical Examiner workload in the Northumbria Police Force area October 2002-January 2003. AB - This force-wide study in the Northumbria Police area provides an audit of the quality and quantity of work carried out by Forensic Medical Examiners (FMEs) over a three month period. Approximately 25% of all arrests during the period of the audit resulted in a request for FME assessment of the detainee. About 79% of consultations were performed to establish fitness to detain, in which over 30% of detainees were intoxicated and over 40% addicted to drugs and/or alcohol. Over 30% of those queried for fitness to detain also required advice and occasionally prescription for medication, both for the management of addiction and the treatment of chronic illness. In comparison to prior work this study found lower rates of mental illness in alcohol and drug dependent populations (13% and 6%, respectively). It is hoped this study will provide information in respect of the specific competencies not only required by future FMEs but also nurses working with detainees within the custody suites. PMID- 16311064 TI - Asthma and air pollution in the Bronx: methodological and data considerations in using GIS for environmental justice and health research. AB - This paper examines methods of environmental justice assessment with Geographic Information Systems, using research on the spatial correspondence between asthma and air pollution in the Bronx, New York City as a case study. Issues of spatial extent and resolution, the selection of environmental burdens to analyze, data and methodological limitations, and different approaches to delineating exposure are discussed in the context of the asthma study, which, through proximity analysis, found that people living near (within specified distance buffers) noxious land uses were up to 66 percent more likely to be hospitalized for asthma, and were 30 percent more likely to be poor and 13 percent more likely to be a minority than those outside the buffers. PMID- 16311065 TI - United snakes. AB - Since their debut in 1987, snakes (active contour models) have become a standard image analysis technique with several variants now in common use. We present a framework called "United Snakes", which has two key features. First, it unifies the most popular snake variants, including finite difference, B-spline, and Hermite polynomial snakes in a consistent finite element formulation, thus expanding the range of object modeling capabilities within a uniform snake construction process. Second, it embodies the idea that the heretofore presumed competing technique known as "live wire" or "intelligent scissors" is in fact complementary to snakes and that the two techniques can advantageously be combined by introducing an effective hard constraint mechanism. The United Snakes framework amplifies the efficiency and reproducibility of the component techniques, and it offers more flexible interactive control while further minimizing user interactions. We apply United Snakes to several different medical image analysis tasks, including the segmentation of neuronal dendrites in EM images, dynamic chest image analysis, the quantification of growth plates in MR images and the isolation of the breast region in mammograms, demonstrating the generality, accuracy and robustness of the tool. PMID- 16311066 TI - Mate choice decisions: the role of facial beauty. AB - For most people, facial beauty appears to play a prominent role in choosing a mate. Evidence from research on facial attractiveness indicates that physical beauty is a sexually selected trait mediated, in part, by pubertal facial hormone markers that signal important biological information about the displayer. Such signals would be ineffective if they did not elicit appropriate cognitive and/or emotional responses in members of the opposite sex. In this article, I argue that the effectiveness of these hormonal displays varies with perceivers' brains, which have been organized by the degree of steroid hormone exposure in the uterus, and activated by varying levels of circulating steroids following puberty. I further propose that the methodology used for examining mate choice decisions has general applicability for determining how cognitive and emotional evaluations enter into decision processes. PMID- 16311067 TI - Oral submucous fibrosis: review on aetiology and pathogenesis. AB - Data from recent epidemiological studies provide overwhelming evidence that areca nut is the main aetiological factor for OSF. A clear dose-dependent relationship was observed for both frequency and duration of chewing areca nut (without tobacco) in the development of OSF. Commercially freeze dried products such as pan masala, Guthka and mawa (areca and lime) have high concentrates of areca nut per chew and appear to cause OSF more rapidly than by self prepared conventional betel quid that contain smaller amounts of areca nut. It is logical to hypothesise that the increased collagen synthesis or reduced collagen degradation as possible mechanisms in the development of the disease. There are numerous biological pathways involved in the above processes and, it is likely that the normal regulatory mechanisms are either down regulated or up regulated at different stages of the disease. Among the chemical constituents, alkaloids from areca nut are the most important biologically whilst tannin may have a synergistic role. These chemicals appear to interfere with the molecular processes of deposition and/or degradation of extracellular matrix molecules such as collagen. In vitro studies on human fibroblasts using areca extracts or chemically purified arecoline support the theory of fibroblastic proliferation and increased collagen formation that is also demonstrable histologically in human OSF tissues. The copper content of areca nut is high and the possible role of copper as a mediator of fibrosis is supported by the demonstration of up regulation of lysyl oxidase in OSF biopsies. It has been postulated that areca nut may also induce the development of the disease by increased levels of cytokines in the lamina propria. Increased and continuous deposition of extracellular matrix may take place as a result of disruption of the equilibrium between matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMP). Current evidence implicates collagen-related genes in the susceptibility and pathogenesis of OSF. The individual mechanisms operating at various stages of the disease-initial, intermediate and advanced-need further study in order to propose appropriate therapeutic interventions. PMID- 16311068 TI - Influence of environmental exposure to PAHs on the susceptibility of lymphocytes to DNA-damage induction and on their repair capacity. AB - The influence of occupational exposure to environmental carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) on DNA damage detected in lymphocytes of exposed people (city policemen) was studied. The cellular susceptibility to the induction of the DNA damage and the repair capacity of exposed donors are presented in comparison with matched controls. Monitoring was performed and blood samples (164 donors) were collected in Prague, Czech Republic, during the winter and summer seasons. The single-cell gel electrophoresis (SCGE) assay with an internal standard was applied to evaluate the DNA damage. A challenging dose of 2Gy of X rays was used to study cellular capacities. In the results of studies of the DNA damage induced in vivo or as an immediate response to the challenging treatment no significant difference was found between exposed and unexposed subgroups. The percentage of non-repaired X-ray-induced DNA damage (residual damage, RD) overall in both seasons was significantly higher in lymphocytes of policemen exposed to c PAHs than in matched controls (RD(T-DNA), %DNA in the comet tail: winter 36.4+/ 22.1 versus 22.7+/-10.8, p < 0.001; summer 47.7+/-22.9 versus 34.7+/-15.2, p < 0.001). The results suggest that occupational exposure to environmental c-PAHs significantly reduces the cellular capacity to repair the DNA damage induced by a challenging treatment. A significant decrease of repair efficiency in donors occupationally exposed to environmental c-PAHs was also observed when subgroups were stratified according to smoking history. In conclusion, our results suggest that environmental exposure to c-PAHs affects the cellular repair processes and can lead to harmful effects hazardous to human health. PMID- 16311069 TI - 4-Hydroxynonenal signalling to apoptosis in isolated rat hepatocytes: the role of PKC-delta. AB - 4-Hydroxynonenal, a significant aldehyde end product of membrane lipid peroxidation with numerous biochemical activities, has consistently been detected in various human diseases. Concentrations actually detectable in vivo (0.1-5 microM) have been shown to up-regulate different genes and modulate various enzyme activities. In connection with the latter aspect, we show here that, in isolated rat hepatocytes, 1 microM 4-hydroxynonenal selectively activates protein kinase C-delta, involved in apoptosis of many cell types; it also induces very early activation of Jun N-terminal kinase, in parallel increasing activator protein-1 DNA-binding activity in a time-dependent manner and triggering apoptosis after only 120 min treatment. These phenomena are likely protein kinase C-delta-dependent, being significantly reduced or annulled by cell co-treatment with rottlerin, a selective inhibitor of protein kinase C-delta. We suggest that 4-hydroxynonenal may induce apoptosis through activation of protein kinase C delta and of Jun N-terminal kinase, and consequent up-regulation of activator protein-1 DNA binding. PMID- 16311070 TI - Serum digoxin concentration and outcomes in women with heart failure: A bi directional effect and a possible effect modification by ejection fraction. AB - BACKGROUND: The association between serum digoxin concentration (SDC) and outcomes in women with heart failure (HF) has not been well studied. AIMS: To test the hypothesis that the effect of digoxin on outcomes in women with HF is bi directional and dependent on SDC, as in men, and is modified by ejection fraction (EF). METHODS: We studied 1366 female participants of the Digitalis Investigation Group trial in whom data on SDC (ng/ml) were available. We calculated adjusted odds ratios (AOR) and Bonferroni-adjusted 97.5% confidence intervals (CI) for various outcomes at a median follow up of 41 months, in all women and stratified by EF 35%. RESULTS: Compared with placebo (26.9%), 40.3% with SDC> or =1.2 (AOR=1.80; CI=1.14-2.86; p=0.004) and 26.6% with SDC 0.5-1.1 (AOR=1.05; CI=0.73 1.51; p=0.762) died. Respective rates for HF-hospitalizations were: placebo (32.8%), SDC> or =1.2 (38.0%) and SDC 0.5-1.1 (25.5%). For women with EF<35% (N=677), SDC 0.5-1.1 lowered odds for HF-hospitalizations (AOR=0.63; CI=0.39 1.00; p=0.026) without increasing odds for death (AOR=0.77; CI=0.47-1.26; p=0.233). In women with EF> or =35% (N=689), SDC 0.5-1.1 had a borderline association with death (AOR=1.58; CI=0.92-2.72; p=0.058) but not with HF hospitalization (AOR=0.95; CI=0.54-1.66; p=0.826). CONCLUSIONS: As in men, in women with HF, digoxin has a bi-directional effect based on SDC, and the beneficial effects were significant only among women with EF<35%. PMID- 16311071 TI - Plasmodium research in the postgenomic era. AB - The complete genomic sequence of Plasmodium falciparum strain 3D7 was published in October 2002. At the Next Steps in Malaria Research meeting in April 2005, the next practical steps were considered and the priorities ranked for postgenomic research in Plasmodium. The high-throughput approaches that will help to answer the major biological questions regarding Plasmodium should, like the genome project itself, build community-shared resources, and efforts must be made to help researchers ready themselves to use the tools that will become available. PMID- 16311072 TI - Strange symptoms in Sneddon's syndrome. PMID- 16311073 TI - Potent vaccine therapy with dendritic cells genetically modified by the gene silencing-resistant retroviral vector GCDNsap. AB - Dendritic cells (DCs) genetically modified to express tumor-associated antigens (TAAs) would be promising tools in cancer immunotherapy. However, the use of retroviral vectors for such modifications is still a challenge because of low transduction efficiency and gene silencing in DCs. We have established an efficient method to prepare such DCs by in vitro differentiation of hematopoietic progenitor cells transduced with chicken ovalbumin (OVA) cDNA via the gene silencing-resistant retroviral vector GCDNsap packaged in vesicular stomatitis virus G protein. When c-KIT(+)/lineage(-) cells were transduced with OVA followed by expansion and differentiation, more than 90% of mature DCs expressed the transgene. Mice inoculated with those cells completely rejected the OVA expressing tumor E.G7-OVA, and the anti-tumor effects were stronger than those observed in mice inoculated with the same number of OVA peptide-pulsed DCs. The mice harbored more cytotoxic T lymphocytes (CTLs) against E.G7-OVA and produced antibody against OVA, suggesting the generation of multiple CTLs recognizing different OVA epitopes and OVA-specific CD4(+) T cells. Successive inoculations of the transduced DCs in a therapeutic setting eradicated preexisting E.G7-OVA and prevented the progression of retransplanted tumors. Thus, this vaccine therapy may represent a potent immunotherapeutic approach for various malignant tumors that express suitable TAAs. PMID- 16311074 TI - Involvement of apoptosis in malathion-induced cytotoxicity in a grass carp (Ctenopharyngodon idellus) cell line. AB - We investigated the role of apoptosis in malathion-induced cytotoxicity in the grass carp (Ctenopharyngodon idellus) cell line ZC-7901. Fish cells were treated with different concentrations of malathion (0.62-95 mg/L), and the IC(50) ranged from 37.94+/-1.93 mg/L for 12 h to 3.04+/-0.27 mg/L for 72 h by the MTT assay. Apoptosis was detected by confocal laser scanning microscopy, transmission electron microscopy, TUNEL reaction, DNA laddering and a flow cytometric PI staining assay. The results demonstrated that apoptosis was involved in the cytotoxic effect of malathion, and that malathion-induced apoptosis occurred in a dose- and time-dependent manner. In addition, the induction of apoptosis by malathion was accompanied by mitochondrial membrane potential (DeltaPsi(m)) disruption, intracellular Ca(2+) elevation, generation of reactive oxygen species (ROS) and ATP depletion. Our investigation suggested that malathion exerts its cytotoxic effects by the induction of apoptosis via a direct effect on the mitochondria. PMID- 16311075 TI - Molecular impact of propiconazole on Daphnia magna using a reproduction-related cDNA array. AB - We have developed a first version cDNA microarray of the cladoceran Daphnia magna. Through Suppression Subtractive Hybridisation PCR (SSH-PCR) 855 life stage specific cDNAs were collected and used to document the toxicological mode of action of the pesticide propiconazole. DNA sequencing analysis revealed gene fragments related to important functional classes such as embryo development, energy metabolism, molting and cell cycle. Major changes in transcription were observed in organisms exposed for 4 and 8 days to 1 microg/mL. After 4 days a 3 fold down-regulation of the gene encoding the yolk protein, vitellogenin, was observed indicating impaired oocyte maturation. Moreover, genes such as a larval specific gene and chaperonin were repressed, whereas the heat shock 90 protein and ATP synthase were induced. Organismal effects clearly confirmed the major molecular findings: at the highest concentration (1 microg/mL) adult growth was significantly (p < 0.05) impaired and increased developmental effects in the offspring could be noted. We have demonstrated the potential of microarray analysis in toxicity screening with D. magna. The use of vitellogenin mRNA as a rapid biomarker of reproductive effects in chronic toxicity studies with cladocerans is suggested. PMID- 16311076 TI - Ancillary study to the PREFER trial: a descriptive study of participants' patterns of self-monitoring--rationale, design and preliminary experiences. AB - BACKGROUND: The treatment of obesity, marked by a high rate of recidivism, lacks strategies that sustain the improved lifestyle behaviors that led to the weight loss. Self-monitoring, a behavioral strategy to increase awareness of one's behavior, is the cornerstone of behavioral treatment in weight loss management. However, adherence to self-monitoring is less than ideal. The actual patterns of self-monitoring of eating behaviors in weight loss treatment programs have not been studied previously; if done, these data could provide information on how self-monitoring patterns affect weight loss and if interventions can be developed that address deficits in self-monitoring. Technology now permits an instrumented paper-and-pencil diary to record the real time of individuals' self-monitoring behaviors (invivodata, inc.), thus providing more information about self-reported behaviors. This technology differs from PDA based products (ex. e-diary, e trials, logpad, PICS) because, to the user, it is no different from a standard paper-and-pencil diary. METHODS: The purposes of this ancillary study are: (1) to describe the electronically recorded patterns of self-monitoring of eating behaviors by weight-loss study participants; (2) compare reported vs. electronically recorded self-monitoring; and (3) examine the relationship between the electronically validated adherence to self-monitoring goals and success in weight loss in a sub-sample of participants in the PREFER study. This paper describes the design, methods and preliminary 6-month results. RESULTS: Preliminary results reveal that participants frequently report recording their eating behavior during times when the binder was not opened. Two examples of data are shown that demonstrate the lack of concordance between the self-reported and the electronically recorded data. CONCLUSIONS: These unobtrusively collected electronic data can lead to the development of strategies that can promote improved adherence to self-monitoring. PMID- 16311077 TI - Carrier frequency of F508del mutation of cystic fibrosis in Indian population. AB - BACKGROUND: Cystic fibrosis (CF) is considered to be very rare in Indian subcontinent. Based on reports of CF in migrants from Indian subcontinent to United Kingdom and United States of America, the prevalence of CF is estimated to be between 1/10,000 and 1/40,000 in this ethnic group. The present study was done to estimate the carrier frequency of F508del mutation among neonates using cord blood samples to reflect the prevalence of CF in the study population. METHODS: 955 mothers delivering at our hospital between December 1999 and November 2000 were enrolled. Cord blood samples were analyzed for F508del mutation using polymerase chain reaction and gel electrophoresis. The frequency of patients homozygous for F508del mutation in the population was estimated using Hardy Weinberg principle. The prevalence of CF was estimated by using the proportion of F508del homozygous cases out of all CF patients, as reported in various studies (19-44%) from Indian subcontinent. RESULTS: Out of 955 cord blood samples, 4 were positive for F508del mutation. The carrier frequency and gene frequency of F508del mutation in the Indian population was calculated to be 1/238 (0.42%) and 1/477 (0.21%), respectively. Frequency of CF patients homozygous for F508del mutation is 1/228,006. The estimated prevalence of CF is 1/43,321 to 1/100,323 in Indian population. CONCLUSION: CF does occur in Indian subcontinent though the prevalence is lesser than the Caucasian population. PMID- 16311078 TI - Altered diaphragm muscle action potentials in Zucker diabetic fatty (ZDF) rats. AB - The Zucker diabetic fatty (ZDF) rat is a model of type 2 diabetes, being characterized by obesity, diabetes, and dyslipidemia. In vitro studies tested the hypothesis that diaphragm muscle from ZDF rats has abnormal resting membrane potential and action potentials, similar to type 1 diabetic rodents. Resting membrane potential was comparable for muscle from ZDF and control rats. Diaphragm from ZDF rats had augmented action potential peak height (92.1 mV versus 82.4 mV, P<0.00001), overshoot (15.6 mV versus 8.1 mV, P<0.001) and area (80.7 mV ms versus 68.6 mV ms, P<0.001) compared with that from controls. Action potential rate of depolarization and repolarization were not affected. The K(+) blocker, 3,4-diaminopyridine, augmented action potential duration and area of muscle from ZDF and controls, but without significant differences between animal groups. These findings in ZDF rats contrast with type 1 diabetic rats, suggesting that isolated hyperglycemia differs from hyperglycemia combined with other metabolic perturbations with respect to diaphragm electrophysiological derangements. PMID- 16311079 TI - Serial respiratory adaptations and an alternate hypothesis of respiratory control in human pregnancy. AB - This study determined the time course of changes in resting and exercising respiratory responses during the first half of human pregnancy, and examined the potential roles of plasma osmolality and the strong ion difference ([SID]) as mediators of pregnancy-induced increases in ventilation. Healthy active women (n = 11) were studied serially from 7 to 22 weeks gestation. Responses were compared with preconception data from 14 subjects (six of whom were tested in early pregnancy), and with late-gestation resting data from 10 additional women. Resting and exercising measurements included ventilation, PaCO2, progesterone, osmolality and [SID]. Results were analyzed using mixed-model linear regression. By 7 weeks gestation, increased ventilation resulted in a very large decrease in PaCO2 at rest and during moderate-intensity exercise. Large correlations (r > 0.5) between resting progesterone and PaCO2 support the traditional theory that circulating progesterone stimulates ventilation during pregnancy. The similar time course of changes and large correlations between raw and delta values of PaCO2 and each of plasma osmolality and [SID] also suggest that both variables may influence respiratory control at rest and during exercise in the first half of pregnancy. Future experiments should continue to explore the hypothesis that osmolality and [SID] contribute to pregnancy-induced respiratory changes. PMID- 16311080 TI - Respiratory changes in a murine model of spontaneous systemic lupus erythematosus. AB - The pathophysiology of systemic lupus erythematosus (SLE) has been very well described in many organs. However, the relation between extracellular matrix changes and lung dynamic mechanical behaviour deserves elucidation. To that end, pulmonary mechanics, lung morphometry and the amount of collagen and elastic fibres in the alveolar septa were analysed in mice with SLE [NZB/W (New Zealand Black/White) F1] and non-diseased NZW mice (control). Static (E(st)) and dynamic (E(dyn)) elastances, difference between dynamic and static elastances (DeltaE), airway resistance (R(aw)) and viscoelastic/inhomogeneous pressure (DeltaP(2)) were determined by the end-inflation occlusion method. Lungs were removed and prepared for histology. E(st), E(dyn), DeltaE and DeltaP(2) were higher in SLE than in control group, while R(aw) was similar in both groups. SLE group showed alveolar collapse and increased amount of elastic and collagen fibres. In conclusion, SLE mice showed an increase in elastic and viscoelastic/inhomogeneous pressures that was accompanied by deposition of collagen and elastic fibres in the alveolar septa. PMID- 16311081 TI - Selective quantitative bioanalysis of proteins in biological fluids by on-line immunoaffinity chromatography-protein digestion-liquid chromatography-mass spectrometry. AB - A quantitative method for the determination of proteins in complex biological matrices has been developed based on the selectivity of antibodies for sample purification followed by proteolytic digestion and quantitative mass spectrometry. An immunosorbent of polyclonal anti-bovine serum albumin (BSA) antibodies immobilized on CNBR agarose is used in the on-line mode for selective sample pretreatment. Next, the purified sample is trypsin digested to obtain protein specific peptide markers. Subsequent analysis of the peptide mixture using a desalination procedure and a separation step coupled, on-line to an ion trap mass spectrometer, reveals that this method enables selective determination of proteins in biological matrices like diluted human plasma. This approach enhances substantially the selectivity compared to common quantitative analysis executed with immunoassays and colorimetry, fluorimetry or luminescence detection. Hyphenation of the immunoaffinity chromatography with on-line digestion and chromatography-mass spectrometry is performed and a completely on line quantification of the model protein BSA in bovine and human urine was established. A detection limit of 170 nmol/l and a quantification limit of 280 nmol/l is obtained using 50 microl of either standard or spiked biological matrix. The model system allows fully automated absolute quantitative mass spectrometric analysis of intact proteins in biological matrices without time consuming labeling procedures. PMID- 16311082 TI - Simple and sensitive determination of free and total morphine in human liver and kidney using gas chromatography-mass spectrometry. AB - We developed a reliable, simple and sensitive method to determine free and total morphine in human liver and kidney, using gas chromatography-mass spectrometry (GC-MS). Free morphine or total morphine obtained by acid hydrolysis from 0.2g tissue sample was extracted using an Extrelut NT column with an internal standard, dihydrocodeine, followed by trimethylsilylation. The derivatized extract was submitted to GC-MS analysis of EI-SIM mode. The calibration curves of morphine in both liver and kidney samples were linear in the concentration range from 0.005 to 5 microg/g. The lower limits of detection of morphine were 0.005 microg/g. This method proved successful when we determined free and total morphine in liver and kidney obtained from an autopsied man who was mis-ingested morphine compound in the hospital, which resulted in the cause of death being morphine intoxication. PMID- 16311083 TI - Randomised clinical trial investigating the effects of combined administration of octreotide and methylglucamine diatrizoate in the older persons with adhesive small bowel obstruction. AB - OBJECTIVE: To investigate the effects of combined administration of octreotide and methylglucamine diatrizoate in the older persons with adhesive small bowel obstruction. PATIENTS AND METHODS: One hundred and sixty-two consecutive patients who had suffered from adhesive intestinal obstruction without clinical evidence of strangulation or gangrene were randomised into two groups, a control group (treated conservatively, n=82) and a contrast group (treated with combined administration of octreotide and methylglucamine diatrizoate, n=80). A laparotomy was performed in both the two groups if symptoms of strangulation developed or the obstruction did not resolve spontaneously after 72 h. RESULTS: Statistically significant rapid reduction in pain score, lower amount of nasogastric drainage, shorter hospital stay, lower operative rate and lower postoperative morbidity were observed in the contrast group. Among the non-operative patients, earlier passage of stool and gas, earlier first oral intake and shorter duration of nasogastric tube placement were significantly more frequently observed in the contrast group. No difference in the rate of readmission was found between the two groups. CONCLUSIONS: Combined administration of octreotide and methylglucamine diatrizoate accelerates resolution of small bowel obstruction by a specific therapeutic effect and is safe for the older persons. PMID- 16311084 TI - Pegylated interferon plus ribavirin for recurrent Hepatitis C infection after liver transplantation in naive and non-responder patients on a stable immunosuppressive regimen. AB - BACKGROUND: Hepatitis C virus recurrence after liver transplantation is universal, leading to chronic hepatitis and cirrhosis. AIMS AND PATIENTS: We evaluated the efficacy and safety of pegylated interferon and ribavirin in 20 patients with recurrent Hepatitis C virus after liver transplantation (10 naive and 10 non-responders to a previous interferon course). METHODS: Treatment consisted of pegylated interferon alfa-2b (1.0 microg/kg once weekly) and ribavirin (600 mg/daily) for at least 6 months. Therapy continued for an additional 6 months only in patients with undetectable serum Hepatitis C virus RNA or >2 log drop from baseline levels. RESULTS: Eleven out of 20 patients (55%) completed 1 year of treatment. Nine patients (45%) had undetectable Hepatitis C virus-RNA at the end of treatment, six of them were naives and three non responders. In all of them, virological response persisted 6 months after discontinuation of therapy, so the sustained virological response rate was 60% in naive patients and 30% in non-responders. CONCLUSIONS: Our results suggest that pegylated interferon plus ribavirin combination therapy may be effective in patients with post-liver transplantation recurrent chronic Hepatitis C, even in those previously non-responders to interferon plus ribavirin. These results need to be confirmed by large studies. PMID- 16311085 TI - Expression and regulation of CCN genes in murine osteoblasts. AB - Members of the CCN family of genes include cysteine-rich 61 (CYR61), connective tissue growth factor (CTGF), nephroblastoma overexpressed (NOV), and Wnt-induced secreted proteins (WISP) 1, 2 and 3. CCN proteins play a role in cell differentiation and function, but their expression and function in skeletal tissue is partially understood. We examined the expression and regulation of CCN genes in primary cultures of murine osteoblasts treated with transforming growth factor beta (TGFbeta), bone morphogenetic protein (BMP)-2, or cortisol. Northern blot analysis revealed the presence of CYR61, CTGF, NOV, and WISP 1 and 2 transcripts in murine osteoblasts, but not WISP 3 transcripts. Northern and Western blot analyses revealed that TGF beta, BMP-2, and cortisol increased CYR61 and CTGF mRNA and protein levels. TGF beta decreased NOV and increased WISP 2 mRNA and protein levels, and TGF beta and BMP-2 increased, whereas cortisol decreased WISP 1 mRNA and protein levels. Nuclear run-on assays revealed that TGF beta, BMP-2 and cortisol enhanced CYR61 and CTGF transcription, TGF beta and BMP 2 induced and cortisol suppressed WISP 1, and TGF beta induced WISP 2 transcription. Suppression of NOV transcription could not be detected due to low control levels. In conclusion, five of the six known CCN genes are expressed by osteoblasts and their transcription is regulated by TGF beta, BMP-2 and cortisol. PMID- 16311086 TI - A systematic review and meta-analysis of the impact of omega-3 fatty acids on selected arrhythmia outcomes in animal models. AB - Epidemiological studies and clinical trials report the beneficial effects of fish or fish oil consumption on cardiovascular disease outcomes including sudden death. We performed a systematic review of the literature on controlled animal studies that assessed the effects of omega-3 fatty acids on selected arrhythmia outcomes. On the basis of predetermined criteria, 27 relevant animal studies were identified; 23 of these were feeding studies, and 4 were infusion studies. Across species, fish oil, eicosapentaenoic acid, and/or docosahexaenoic acid appear to have beneficial effects on ventricular tachycardia (VT) and fibrillation (VF) in ischemia- but not reperfusion-induced arrhythmia models; no effect on the incidence of death and infarct size; and inconsistent results with regard to arrhythmia score, VF threshold, ventricular premature beats or length of time in normal sinus rhythm, compared to omega-6, monounsaturated, or saturated fatty acids, and no treatment controls. In a meta-analysis of 13 studies using rat models, fish oil but not alpha-linolenic acid supplementation showed a significant protective effect for ischemia- and reperfusion-induced arrhythmias by reducing the incidence of VT and VF. It is not known whether omega-3 fatty acid supplementation has antiarrhythmic effects in other disease settings not related to ischemia. PMID- 16311087 TI - Hyperinsulinemia predicts hepatic fat content in healthy individuals with normal transaminase concentrations. AB - Although the prevalence of insulin resistance (IR) and compensatory hyperinsulinemia (CH) is increased in patients with nonalcoholic fatty liver disease, the role of IR/CH in regulation of hepatic fat content in healthy volunteers with normal concentrations of alanine transaminase (ALT) has not been defined. To address this issue, hepatic fat content was quantified by ultrasound in 69 (30 men, 39 women) healthy individuals, without known risk factors for liver disease and with plasma ALT concentrations of less than 30 U/L. Experimental variables quantified included body mass index, waist circumference, systolic and diastolic blood pressures, and fasting plasma glucose, fasting plasma insulin (FPI), and lipid concentrations. Subjects were classified as having no (55%), mild (27%), or moderate to severe (18%) hepatic steatosis on the basis of the ultrasound results. Statistically significant (P < .05-.001) correlations (Spearman rho values) existed between liver fat content and ALT (0.26), body mass index (0.52), waist circumference (0.50), systolic blood pressure (0.28), diastolic blood pressure (0.27), fasting plasma glucose (0.47), FPI (0.56), triglycerides (0.30), and high-density lipoprotein cholesterol ( 0.35). Multivariate general discriminant analysis and multiple linear regression analysis indicated that FPI was the only independent predictor (P < .001) of both liver fat content and ALT concentrations. Fasting plasma insulin (a surrogate estimate of IR/CH) predicts hepatic fat content and ALT in healthy volunteers with normal transaminase concentrations, independently of the other anthropometric and metabolic variables measured. PMID- 16311088 TI - Prevention of hyperglycemia in Zucker diabetic fatty rats by exercise training: effects on gene expression in insulin-sensitive tissues determined by high density oligonucleotide microarray analysis. AB - Exercise training (ET) causes metabolic improvement in the prediabetic and diabetic states. However, only little information exists on the changes to ET at the transcriptional level in insulin-sensitive tissues. We have investigated the gene expression changes in skeletal muscle, liver, fat, and pancreatic islets after ET in male Zucker diabetic fatty (ZDF) rats. Eighteen ZDF rats (7 weeks old) were divided in a control and ET group. Exercise was performed using a motorized treadmill (20 m/min 1 hour daily for 6 days a week). Blood glucose, weight, and food intake were measured weekly. After 5 weeks, blood samples, soleus muscle, liver, visceral fat (epididymal fat pads), and islet tissue were collected. Gene expression was quantified with Affymetrix RG-U34A array (16 chips). Exercise training ameliorates the development of hyperglycemia and reduces plasma free fatty acid and the level of glucagon-insulin ratio (P < .05). In skeletal muscle, the expression of 302 genes increased, whereas that of 119 genes decreased. These changes involved genes related to skeletal muscle plasticity, Ca(2+) signals, energy metabolism (eg, glucose transporter 1, phosphorylase kinase), and other signaling pathways as well as genes with unknown functions (expressed sequence tags). In the liver, expression of 148 genes increased, whereas that of 199 genes decreased. These were primarily genes involved in lipogenesis and detoxification. Genes coding for transcription factors were changed in parallel in skeletal muscle and liver tissue. Training did not markedly influence the gene expression in islets. In conclusion, ET changes the expression of multiple genes in the soleus muscle and liver tissue and counteracts the development of diabetes, indicating that ET-induced changes in gene transcription may play an important role en the prevention of diabetes. PMID- 16311089 TI - Effect of aromatic ring-containing drugs on carnitine biosynthesis in rats with special regard to p-aminomethylbenzoic acid. AB - Secondary carnitine deficiencies are associated with metabolic disorders or may be the consequence of the side effects of some drugs. The mechanisms may be either a facilitated urinary excretion or an inhibited biosynthesis. Based on our earlier findings with drugs and benzoic acid analogue metabolites, in the present study, we studied the possible inhibitory effect of some benzoic acid analogue drugs. In the pathway of carnitine biosynthesis, we tested the last step, the hydroxylation of gamma-butyrobetaine (Bu) to carnitine in the liver. (Liver is the only organ in rats where this step takes place.) Of the 5 tested compounds, the p-aminomethylbenzoic acid (PAMBA) was found to be inhibitory. In tracer experiments with radioactive Bu, PAMBA (a single injection of 1.2 mmol/kg) reduced the conversion of [Me-(3)H]Bu to [Me-(3)H]carnitine from 62.6% +/- 5.11% to 46.8% +/- 5.02% (means +/- SEM, P < .02). This single dose also markedly reduced the conversion of loading amount of exogenous unlabeled Bu, as measured by enzymatic analysis of carnitine. The conversion of endogenous Bu was also hampered by long-term administration of PAMBA, as indicated by increased Bu and decreased carnitine levels. Furthermore, single injection of PAMBA markedly reduced the Glu level in the liver from 2.87 +/- 0.17 to 1.42 +/- 0.11 mumol/g (P < .001). Trying to get closer to a mechanism by which the flux through the Bu hydroxylase was depressed, we supposed that alfa-ketoglutarate (alpha-KG), an obligatory cofactor of the enzyme, was also be depressed. It was expected because alpha-KG is a reversible copartner of l-glutamate through the Glu-dehydrogenase reaction. We found that PAMBA reduced the alpha-KG level from 207 +/- 17.5 to 180 +/- 19.1 nmol/g (means +/- SEM, P < .02). Considering the conditions of the enzyme in vitro and in vivo, this decrease may contribute to the decreased in vivo flux through the butyrobetaine hydroxylase enzyme. PMID- 16311090 TI - NO-1886 (ibrolipim), a lipoprotein lipase activator, increases the expression of uncoupling protein 3 in skeletal muscle and suppresses fat accumulation in high fat diet-induced obesity in rats. AB - Although the lipoprotein lipase (LPL) activator NO-1886 shows antiobesity effects in high-fat-induced obese animals, the mechanism remains unclear. To clarify the mechanism, we studied the effects of NO-1886 on the expression of uncoupling protein (UCP) 1, UCP2, and UCP3 in rats. NO-1886 was mixed with a high-fat chow to supply a dose of 100 mg/kg to 8-month-old male Sprague-Dawley rats. The animals were fed the high-fat chow for 8 weeks. At the end of the administration period, brown adipose tissue (BAT), mesenteric fat, and soleus muscle were collected and levels of UCP1, UCP2, and UCP3 messenger RNA (mRNA) were determined. NO-1886 suppressed the body weight increase seen in the high-fat control group after the 8-week administration (585 +/- 39 vs 657 +/- 66 g, P < .05). NO-1886 also suppressed fat accumulation in visceral (46.9 +/- 10.4 vs 73.7 +/- 14.5 g, P < .01) and subcutaneous (43.1 +/- 18.1 vs 68.9 +/- 18.8 g, P < .05) tissues and increased the levels of plasma total cholesterol and high-density lipoprotein cholesterol in comparison to the high-fat control group. In contrast, NO-1886 decreased the levels of plasma triglycerides, nonesterified free fatty acid, glucose, and insulin. NO-1886 increased LPL activity in soleus muscle (0.082 +/- 0.013 vs 0.061 +/- 0.016 mumol of free fatty acid per minute per gram of tissue, P < .05). NO-1886 increased the expression of UCP3 mRNA in soleus muscle 3.14-fold (P < .01) compared with the high-fat control group without affecting the levels of UCP3 in mesenteric adipose tissue and BAT. In addition, NO-1886 did not affect the expression of UCP1 and UCP2 in BAT, mesenteric adipose tissue, and soleus muscle. In conclusion, NO-1886 increased the expression of UCP3 mRNA and LPL activity only in skeletal muscle. Therefore, a possible mechanism for NO-1886's antiobesity effects in rats may be the enhancement of LPL activity in skeletal muscle and the accompanying increase in UCP3 expression. PMID- 16311091 TI - Effects of thorough mastication on postprandial plasma glucose concentrations in nonobese Japanese subjects. AB - Thorough mastication has the potential to affect postprandial plasma glucose concentrations by improving digestibility and absorption of nutrients. To evaluate the effects of mastication on postprandial plasma glucose concentration, we compared usual and thorough mastication in subjects with normal glucose tolerance (NGT group, n = 16) and subjects predisposed to type 2 diabetes (first degree relatives of type 2 diabetic patients, subjects with impaired glucose tolerance, and type 2 diabetic patients) (predisposed group, n = 10) in a crossover trial of 52 test meals. Plasma glucose and serum insulin concentrations were measured for 3 hours postprandially, and the insulinogenic index (the ratio of incremental serum insulin to plasma glucose concentration during the first 30 minutes after meal) was calculated. In the NGT group, thorough mastication reduced the postprandial plasma glucose concentration at 90 minutes (5.8 +/- 0.3 vs 6.5 +/- 0.4 mmol/L, P < .05) and 120 minutes (5.4 +/- 0.2 vs 6.3 +/- 0.4 mmol/L, P < .05) and the area under the curve (AUC) from -15 to 180 minutes (19.1 +/- 0.6 vs 20.6 +/- 0.8 [mmol . L]/h, P < .05) without an increase in the AUC for insulin. In the predisposed group, thorough mastication significantly augmented plasma glucose and serum insulin concentrations and the AUCs compared with usual mastication. Thorough mastication elicited a significantly higher insulinogenic index than usual mastication in the NGT group (205.0 +/- 27.6 vs 145.6 +/- 17.7 pmol/mmol, P < .05), whereas the predisposed group showed significantly less early-phase insulin secretion than the NGT group. In the NGT group the postprandial plasma glucose concentration upon thorough mastication of meal was significantly lower, most probably because of the potentiation of early-phase insulin secretion. In the subjects predisposed to type 2 diabetes, thorough mastication did not potentiate early-phase insulin secretion and elicited a higher postprandial plasma glucose concentration. PMID- 16311092 TI - Fatty acid-induced changes in vascular reactivity in healthy adult rats. AB - Dietary fatty acids (FAs) are known to modulate endothelial dysfunction, which is the first stage of atherosclerosis. However, their exact role in this initial phase is still unclear. The effects of isolated or combined (by 2) purified FAs from the main FA families were studied on the vascular response of isolated thoracic aorta in healthy rats to get a better understanding of the mechanisms of action of dietary FAs in regulating vascular endothelial function. Cumulative contraction curves to phenylephrine and relaxation curves to carbachol and then to sodium nitroprusside were obtained in the absence or presence of the FAs studied allowing endothelium-dependent and endothelium-independent ability of the smooth muscle to relax to be assessed in each experimental group. The endothelium dependent vasodilator response to carbachol was lowered by eicosapentaenoic acid, whereas it was not altered either by docosahexaenoic acid alone or by combined eicosapentaenoic acid-docosahexaenoic acid, oleic acid, or stearic acid, and it was increased by linoleic acid (LA). A decreased phenylephrine-induced contraction was observed after incubation with arachidonic acid and with stearic acid. On the other hand, the endothelium-dependent relaxation was reduced by the addition of combined LA-arachidonic acid and LA-oleic acid. In conclusion, these data point out the differential effects of different types of FAs and of FAs alone vs combined on vascular reactivity. The complex nature of these effects could be partially linked to metabolic specificities of endothelial cells and to interactions between some FAs. PMID- 16311093 TI - Effect of postmenopause and hormone replacement therapy on serum adiponectin levels. AB - Little is known about the effects of menopause and hormone replacement therapy (HRT) on adiponectin production. The objectives of the study were to compare levels of serum adiponectin in post- and premenopausal women, to test whether adiponectin levels are related to endogenous estradiol and sex hormone-binding globulin (SHBG) levels, to determine whether HRT influences serum adiponectin, and to investigate relationships of adiponectin levels with cardiovascular risk factors. One hundred four women matched for body mass index were enrolled in this study, and among them were 34 postmenopausal HRT nonusers, 34 postmenopausal HRT users, and 36 premenopausal healthy women with regular menstrual cycles. We evaluated waist circumference and waist-to-hip ratio (WHR) in each women. Serum was assayed for adiponectin, estradiol, SHBG, triglycerides (TGs), total cholesterol, high-density lipoprotein cholesterol, and fasting glucose levels. Post- and premenopausal women showed no significant differences in adiponectin and SHBG concentrations. There were no differences in serum adiponectin levels between postmenopusal HRT nonusers and users; however, SHBG concentrations were higher in HRT users. The simple linear regression analyses of all studied women indicated that serum adiponectin was negatively correlated with body mass index, waist circumference, WHR, and TG levels. Positive correlation was observed between adiponectin and high-density lipoprotein cholesterol as well as between adiponectin and SHBG levels. There were no relationships between adiponectin and estradiol levels in all studied women and among subgroups. Multiple regression analysis showed that WHR and TG were significant independent predictors of serum adiponectin. In conclusion, serum adiponectin levels are not influenced by menopausal status or serum estradiol levels. Exogenous estrogen treatment does not significantly affect serum adiponectin concentrations. PMID- 16311094 TI - Variant clinical courses of 2 patients with neonatal intrahepatic cholestasis who have a novel mutation of SLC25A13. AB - Deficiency of citrin due to mutations of the SLC25A13 gene causes not only adult onset type II citrullinemia, but also neonatal intrahepatic cholestasis. Neonatal intrahepatic cholestasis is a self-limiting condition and spontaneously disappears by 12 months of age without special treatment. The natural history of patients with SLC25A13 mutations is not clear. Two patients with infantile hepatic dysfunction were found to have a novel mutation of the SLC25A13 gene. DNA analyses of SLC25A13 disclosed that the first patient was a compound heterozygote for the Ex16+74_IVS17-32del516 (del516-Ex16/IVS17) and IVS11+1G-->A mutations and the second one a homozygote for the del516-Ex16/IVS17 mutation. It is predicted that the 516-base pair deletion mutation leads to a frameshift from codons 556 to 564, a premature termination at codon 565, and a truncated form of the citrin protein (normal, 675 amino acids). The first patient had disseminated intravascular coagulation associated with hepatic dysfunction in the neonatal period. The other patient had persistent cholestatic jaundice and underwent an operation to rule out bile duct atresia. Without specific treatment, both patients had a favorable clinical course. In conclusion, citrin deficiency resulting from the mutation of SLC25A13 presented variant clinical courses, followed by hypercitrullinemia and intrahepatic cholestasis in infancy. The conditions in the patients were self-limiting and spontaneously disappeared. PMID- 16311095 TI - Influence of cyclosporine on low-density lipoprotein uptake in human lymphocytes. AB - Low-density lipoprotein (LDL) levels are often elevated in renal transplant recipients, and cyclosporine (CsA) therapy in these patients has been implicated. Cardiovascular disease is the major cause of mortality in transplant recipients, and alterations of lipid metabolism represent a common risk factor. The role of CsA on LDL metabolism is still partially defined. The aim of the study was to evaluate the LDL receptor uptake of CsA-transported LDL (CsA-LDL) compared with normal LDL in normal and CsA-treated lymphocytes. Forty-seven healthy unrelated subjects and 6 CsA-treated patients were consecutively enrolled as donors of lymphocytes to measure receptor-mediated LDL metabolism. Normal LDL and CsA-LDL were isolated from blood donors and from patients under CsA immunosuppressive therapy, respectively. Lipoproteins were labeled with a fluorochrome, and LDL receptor uptake was measured by flow cytometry. Normal LDL uptake was 13.95% +/- 4.5%, whereas CsA-LDL uptake was 32.47% +/- 10.84% (P < .001) in healthy lymphocytes. In CsA-treated lymphocytes, normal LDL uptake was 7.48% +/- 2.32% vs 12.49% +/- 2.44% CsA-LDL (P < .01). Lymphocytes of every subject showed at least a 2-fold increased uptake of CsA-LDL vs normal LDL. Our data show that CsA-LDL is internalized more than normal LDL via the LDL receptor in both human healthy and CsA-treated lymphocytes. CsA-treated lymphocytes, in comparison to normal lymphocytes, exhibit a reduced LDL receptor activity. PMID- 16311096 TI - Does androgen excess contribute to the cardiovascular risk profile in postmenopausal women with type 2 diabetes? AB - The purpose of this study was to determine if postmenopausal women with type 2 diabetes have clinical and biochemical evidence of androgen excess as a potential contributor to an increase in risk for coronary heart disease when compared with women without diabetes. Fasting glucose, insulin, lipids, sex hormone-binding globulin (SHBG), and sex steroids (from pooled samples) (total testosterone and free testosterone [non-SHBG-T], androstenedione [A-dione], total estrogens) were measured at baseline in 16 postmenopausal women with type 2 diabetes treated with diet or a sulfonylurea and 17 age-matched controls. Measurements of glucose, insulin, and sex steroids were repeated at hourly intervals for 3 hours after oral glucose administration. Hirsutism scores and insulin sensitivity (homeotasis model assessment [HOMA] insulin [SI]) were obtained. Women with type 2 diabetes were more hyperglycemic, hyperinsulinemic, and insulin-resistant (HOMA SI, 46.7 +/- 7.0 vs 12.9 +/- 2.0, P < .001), and had higher total to high-density lipoprotein cholesterol (TC/HDL) ratios, lower SHBG (20.8 +/- 3.5 vs 59.3 +/- 14.4 nmol/L, P < .05), higher non-SHBG-T (0.225 +/- 0.025 vs 0.135 +/- 0.021 nmol/L, P < .05), and higher hirsutism scores (1.1 +/- 0.3 vs 0.3 +/- 0.2, P = .004) than those without diabetes. No changes in sex steroids occurred after the oral glucose challenge. HOMA SI and area under the curve for glucose correlated significantly with SHBG (r = -0.42), non-SHBG-T (r = 0.40), and TC/HDL (r = 0.41) (all P < .05) in the combined groups. Postmenopausal women with type 2 diabetes have both clinical and biochemical evidence of androgen excess that may contribute to more adverse cardiovascular risk profiles. PMID- 16311097 TI - Sex-specific effect of APOAV variant (Val153>Met) on plasma levels of high density lipoprotein cholesterol. AB - The importance of the APOAV gene for the determination of plasma triglyceride levels has been suggested by creations of transgenic and knockout mice and confirmed in population studies. We examined whether the newly detected APOAV variant is associated with plasma lipid levels and risk of myocardial infarction (MI). APOAV polymorphism (Val153>Met) was genotyped in 1191 males and 1368 females representatively selected from the Czech population. Lipid levels were analyzed in 1997 and 2001 in all individuals. Subsequently, we have analyzed the genotype frequencies of APOAV polymorphism in 435 male patients with MI. Val153>Met variation in the APOAV gene affects the plasma high-density lipoprotein cholesterol levels showing a higher level in Val/Val homozygotes than in Met carriers in both years (1.51 +/- 0.36 and 1.52 +/- 0.37 mmol/L compared with 1.42 +/- 0.33 and 1.39 +/- 0.35 mmol/L, P < .01). This association has been observed in females but not in males. Other analyzed lipid parameters (total cholesterol, low-density lipoprotein cholesterol, and triglycerides) have not been associated with APOAV Val153>Met variant. In a group of patients with MI, the frequency of the Met153 carriers was not significantly different from the male population sample (6.5% vs 6.4%). Val153>Met variation in the APOAV gene plays a sex-specific role in genetic determination of plasma high-density lipoprotein cholesterol levels, but does not influence risk of MI in males. PMID- 16311098 TI - Inhibition of the phosphatidylinositol 3'-kinase signaling pathway leads to decreased insulin-stimulated adiponectin secretion from 3T3-L1 adipocytes. AB - Adiponectin is a protein secreted by adipocytes, which modulates insulin resistance and is thought to confer protection from atherosclerosis. Decreased circulating adiponectin is seen in states of insulin resistance, yet the cause of this decrease remains unclear. We investigated the role of insulin in adiponectin secretion and the effect of selective insulin resistance on insulin-stimulated adiponectin secretion by 3T3-L1 adipocytes. Inhibition of the phosphatidylinositol 3'-kinase (PI3K) insulin-signaling pathway was induced with wortmannin (WT) or with a kinase-inactive Akt adenoviral construct (Akt-KD), and inhibition of the mitogen-activated protein kinase pathway was induced with PD98059 or with a dominant-negative ras adenoviral construct (DNras). The PI3K pathway was activated with a constitutively active Akt adenoviral construct (Akt myr). Adiponectin was measured by Western blot, and adiponectin messenger RNA (mRNA) levels were determined by real-time reverse transcription-polymerase chain reaction. Insulin treatment increased adiponectin secretion and decreased intracellular adiponectin. Treatment with 100 nmol/L insulin for 24 hours resulted in a 78% increase in secreted adiponectin (P < .05). Insulin had no effect on adiponectin mRNA. WT or Akt-KD, but not PD98059 or DNras, inhibited insulin-stimulated adiponectin secretion (P < .05). Activation of the PI3K pathway resulted in increased insulin-independent adiponectin secretion. Inhibition of the PI3K- or mitogen-activated protein kinase-dependent pathway decreased adiponectin mRNA by 50% (P < .01). We demonstrate a decrease in insulin stimulated adiponectin secretion with selective inhibition of the PI3K pathway. These results suggest a mechanism for the observed decreased adiponectin levels associated with insulin resistance, when defects in the PI3K-dependent insulin signaling pathway lead to decreased adiponectin production, inadequate adiponectin secretion, and therefore low circulating adiponectin levels. PMID- 16311099 TI - Spectrum of insulin sensitivity in the Korean population. AB - The aims of the present study were to (1) examine the range of values for insulin sensitivity measures such as fasting serum insulin, homeostasis model assessment (HOMA), and quantitative insulin sensitivity check index (QUICKI) and (2) to identify cutoffs for indirect indexes of insulin sensitivity such as insulin, HOMA, and QUICKI that confer increased risk of metabolic syndrome in a large sample of Korean adults. The total number of study subjects involved was 83186. All of them presented for a routine health status checkup at the Kangbuk Samsung Hospital between January 2003 and December 2004, and none of them was currently taking medication for hypertension, diabetes, or dyslipidemia. We used 3 measures of insulin sensitivity: the fasting serum insulin, the HOMA, and the QUICKI. The age- and sex-adjusted prevalence of metabolic syndrome was examined by tenths of the distribution of each index of insulin. The fasting serum insulin, HOMA, and QUICKI were compared by using receiver operating characteristic curves. The fasting serum insulin ranged from 1.71 to 70.40 microU/mL, with the 25th percentile = 5.97, the median = 7.69, and the 75th percentile = 9.82. The HOMA ranged from 0.34 to 17.72, with the 25th percentile = 1.33, the median = 1.74, and the 75th percentile = 2.27. The QUICKI ranged from 0.112 to 0.202, with the 25th percentile = 0.146, the median = 0.152, and the 75th percentile = 0.158. The insulin, HOMA, and QUICKI values at the point on the receiver operating characteristic curve closest to the ideal of 100% sensitivity and 100% specificity for detecting the presence of metabolic syndrome were 9.7 microU/mL, 2.43, and 0.145, respectively. In conclusion, these findings describe the spectrum of insulin sensitivity in Korean adults. This study is the first attempt to determine cutoff values for indirect indexes of insulin sensitivity such as insulin, HOMA, and QUICKI that confer an increased risk of metabolic syndrome. These findings may be useful for evaluating insulin resistance, particularly in epidemiological studies. PMID- 16311100 TI - Comparison of the acute response to meals enriched with cis- or trans-fatty acids on glucose and lipids in overweight individuals with differing FABP2 genotypes. AB - Trans-fatty acids have been implicated as a risk factor for cardiovascular disease and diabetes. In addition, a polymorphism at codon 54 (Ala54Thr) in the fatty acid-binding protein 2 (FABP2) gene has been suggested to modify an interaction between dietary fat and insulin sensitivity. We examined the postprandial metabolic profiles after meals enriched with C18:1trans- relative to a similar meal with C18:1cis-fatty acid in individuals who were either FABP2 Ala54 homozygotes or Thr54 carriers. Moderately overweight men and women ate 2 breakfast test meals, separated by 1 week, each providing 40% of their daily energy requirement and containing 50% of energy as fat. In one meal, 10% of energy was from C18:1trans, and in the other meal, the C18:1trans was replaced with C18:1cis. Metabolic parameters were assessed during an 8-hour period. Insulin and C-peptide levels increased more after the C18:1trans meal, and this was associated with a greater fall in free fatty acids. Postprandial glucose levels and oxidation of fatty acids and carbohydrate were not different between the 2 test meals. The Thr54 allele for FABP2 increased the rise in postprandial glucose but not triacylglycerols. Fractional triacylglycerol synthetic rates were higher after consumption of the C18:1trans meal relative to the C18:1cis meal only in Thr54 carriers. These data show that a single meal enriched with C18:1trans-fatty acids can significantly increase insulin resistance, and that in the presence of the FABP2 Thr54 allele, may contribute to increased partitioning of glucose to triacylglycerols and insulin resistance. PMID- 16311101 TI - Selective angiotensin II receptor antagonism enhances whole-body insulin sensitivity and muscle glucose transport in hypertensive TG(mREN2)27 rats. AB - Essential hypertension is frequently associated with insulin resistance of skeletal muscle glucose transport, and angiotensin II (ANGII) can contribute to the pathogenesis of both conditions. The male heterozygous TG(mREN2)27 rat (TGR) harbors the mouse transgene for renin, exhibits local tissue elevations in ANGII and is an excellent model of both hypertension and insulin resistance associated with defective insulin signaling. The present study was designed to assess the specific role of ANGII in the insulin resistance of the male heterozygous TGR. TGRs were treated with either vehicle or the ANGII (AT(1)-specific) receptor antagonist, irbesartan (50 mg/kg body weight), for 21 consecutive days. Compared with vehicle-treated TGRs, whole-body insulin sensitivity was increased 35% (P < .05) in the irbesartan-treated group, and insulin-mediated glucose transport was increased (P < .05) in both type IIb epitrochlearis (80%) and type I soleus (59%) muscles after irbesartan treatment. Moreover, glycogen synthase activation due to insulin was increased 58% (P < .05) in the soleus of the irbesartan-treated TGRs. However, no significant improvements were observed for functionality of insulin signaling elements (tyrosine phosphorylation of insulin receptor and insulin receptor substrate 1 [IRS1], IRS1 associated with the p85 regulatory subunit of phosphatidylinositol 3'-kinase, and Ser473 of Akt) in muscle of irbesartan treated animals, except for a 25% increase (P < .05) in IRS1 tyrosine phosphorylation in soleus. Collectively, these data indicate that the improvements in whole-body and skeletal muscle insulin action after long-term antagonism of ANGII action in TGRs occur independently of modulation of the functionality of these insulin-signaling elements. PMID- 16311102 TI - Effect of a 2-day very low-energy diet on skeletal muscle insulin sensitivity in obese type 2 diabetic patients on insulin therapy. AB - This study investigates the molecular mechanisms underlying the blood glucose lowering effect of a 2-day very low-energy diet (VLED, 1883 kJ/d) in 12 obese (body mass index, 36.3 +/- 1.0 kg/m2 [mean +/- SEM]) type 2 diabetic (HbA(1C) 7.3% +/- 0.4%) patients simultaneously taken off all glucose-lowering therapy, including insulin. Endogenous glucose production (EGP) and glucose disposal ([6,6 2H2]-glucose) were measured before and after the VLED in basal and hyperinsulinemic (40 mU/m2 per minute) euglycemic conditions. Insulin signaling and expression of GLUT-4, FAT/CD36, and triglycerides were assessed in muscle biopsies, obtained before the clamp and after 30 minutes of hyperinsulinemia. Fasting plasma glucose decreased from 11.3 +/- 1.3 to 10.3 +/- 1.0 mmol/L because of a decreased basal EGP (14.2 +/- 1.0 to 11.9 +/- 0.7 micromol/kg per minute, P = .009). Insulin-stimulated glucose disposal did not change. No diet effect was found on the expression of the insulin receptor and insulin receptor substrate-1 or on phosphatidylinositol 3'-kinase activity, or on FAT/CD36 expression pattern, GLUT-4 translocation, or triglyceride distribution in either the basal or insulin stimulated situation. Unexpectedly, basal PKB/Akt phosphorylation on T308 and S473 increased after the diet, at equal protein expression. In conclusion, a 2 day VLED lowers fasting plasma glucose via a decreased basal EGP without an effect on glucose disposal. Accordingly, no changes in activation of phosphatidylinositol 3'-kinase, triglyceride distribution, FAT/CD36 expression, and GLUT-4 translocation were found in skeletal muscle biopsies. PMID- 16311103 TI - Captopril does not affect reflex increases in adrenal or lumbar sympathetic nerve activity to hypoglycemia in rats. AB - Blockade of angiotensin II (ANGII) receptors or converting enzyme inhibition attenuates reflex increases in epinephrine during insulin-induced hypoglycemia. Because ANGII receptors are found in several sites within the central nervous system, the aim of this study was to examine whether acute captopril attenuates the reflex increase in adrenal preganglionic sympathetic nerve activity (SNA) induced by hypoglycemia. We infused vehicle (control) or insulin (30 U/kg IV) in anesthetized rats or in rats pretreated with captopril (Cap-insulin; 2.5 mg/kg, then 1 mg/kg per hour IV) while measuring hemodynamics and SNA from adrenal preganglionic, adrenal postganglionic, and lumbar sympathetic nerves. Hypoglycemia elicited similar adrenal preganglionic SNA increases in insulin treated (260% +/- 31% from 100% baseline) and Cap-insulin-treated (255% +/- 34%) rats. Likewise, increases in adrenal postganglionic SNA and lumbar SNA were equivalent in the insulin and Cap-insulin groups. Hypoglycemia also elicited a tachycardia in insulin-treated rats that was attenuated in Cap-insulin-treated rats, and corresponding blood pressure decreases in insulin rats were enhanced in Cap-insulin-treated rats. Thus, blockade of ANGII formation by captopril did not affect hypoglycemia-induced activation of adrenal preganglionic SNA, indicating that the renin-angiotensin systems in the brain and spinal cord do not modulate increases in adrenal SNA during hypoglycemia. PMID- 16311104 TI - Continually high insulin levels impair Akt phosphorylation and glucose transport in human myoblasts. AB - Chronic hyperinsulinemia is both a marker and a cause for insulin resistance. This study analyzes the effect of long-term exposure to high insulin levels on the insulin-signaling pathway and glucose transport in cultured human myoblasts. Human myoblasts were grown in the presence of low (107 pmol/L, SkMC-L) or high (1430 pmol/L, SkMC-H) insulin concentrations for 3 weeks. Glucose transport, insulin receptor (IR), and IR substrate 1 (IRS1) phosphorylation, phosphatidylinositol 3'-kinase (PI3K) activity, as well as Akt-Ser473 phosphorylation have been investigated at the end of the incubation period and after a further short-term insulin stimulation. At the end of the incubation period, IR, IRS1, p85/PI3K, Akt, and GLUT4 protein expression levels were similar in both culture conditions. Basal glucose transport was similar in SkMC-L and SkMC-H, but after short-term insulin stimulation significantly increased (P < .01) only in SkMC-L. IR binding was down-regulated in SkMC-H (P < .01), but IR and IRS1 tyrosine phosphorylation and PI3K activity were significantly higher (P < .01) in SkMC-H than SkMC-L. Despite increased PI3K activation, Akt-Ser473 phosphorylation was similar in SkMC-L and SkMC-H. After a short-term insulin stimulation (10 nmol/L insulin for 10 minutes), IR and IRS1 tyrosine phosphorylation, PI3K activation, and Akt-Ser473 phosphorylation significantly increased (P < .01 and P < .05 for Akt) in SkMC-L but not in SkMC-H. Serine phosphorylation of IRS1 was similar in SkMC-L and SkMC-H. Moreover, in the SkMC H, insulin stimulation was associated with the inhibition of IRS1 tyrosine dephosphorylation (P < .05). In summary, continuous exposure of cultured myoblasts to high insulin levels induces a persistent up-regulation of IR, IRS1, and PI3K activity associated with the demodulation of insulin signaling. Moreover, the impairment of the insulin-signaling steps between PI3K and Akt is concomitant with the desensitization of glucose transport. These alterations may contribute to the derangement insulin-signaling pathway states of hyperinsulinemia such as obesity and type 2 diabetes. PMID- 16311106 TI - Osteosarcoma: basic science and clinical implications. AB - Current therapy for osteosarcoma successfully treats 60% to 70% of patients. Attempts to identify patients who will respond poorly to therapy has focused on the use of new biologic markers or microarray cluster analysis. New potential therapeutic targets, including growth factors, chemokines, transcription factors, and angiogenic factors, are being evaluated for their roles in osteosarcoma. These new targets may provide mechanisms to treat the patients who would respond poorly to standard therapy. PMID- 16311107 TI - Recent advances in the basic science of chondrosarcoma. AB - Chondrosarcoma are a heterogeneous group of tumors whose treatment is limited to surgical removal. Molecular genetic analysis of these tumors has revealed some of the abnormalities responsible for the traits of the malignant phenotype. PMID- 16311108 TI - Advances in chemotherapy for patients with extremity soft tissue sarcoma. AB - Doxorubicin-based chemotherapy does not appear to offer a survival benefit to patients who have high-risk primary extremity soft tissue sarcomas, whereas ifosfamide-based chemotherapy does. This benefit is likely histology- and size specific. Until a less toxic targeted systemic therapy is developed, treatment with ifosfamide should be strongly considered in patients who have high-risk primary extremity soft tissue sarcomas. PMID- 16311109 TI - Pigmented villonodular synovitis. AB - Pigmented villonodular synovitis is a benign proliferative lesion of the synovium with unclear etiology. It is a locally aggressive lesion that may invade and destroy surrounding bone and soft tissues and represents a high rate of recurrence despite aggressive treatment modalities. This article describes the new developments in etiology and outlines current approaches for diagnosis and treatment. PMID- 16311110 TI - Giant cell tumor of bone. AB - Giant cell tumor is a common benign bone tumor that possesses specific features including location at the end of long bone, a strong tendency toward local recurrence, and the rare capacity to metastasize to the lungs. Preferred treatment usually consists of extensive curettage and filling of the cavity with bone graft or cement. Debate still exists about the usefulness of local adjuvant treatment. Functional outcome is usually very good. PMID- 16311111 TI - Desmoid tumors and current status of management. AB - Desmoid tumors, also known as aggressive fibromatosis, are rare fibroblastic tumors that exhibit a wide range of local aggressiveness, from largely indolent to locally destructive. Understanding of the pathogenesis and the great heterogeneity in the natural history of desmoid tumors is invaluable to the development of therapeutic strategies. The optimal treatment protocol has not yet been established and, in many cases, a multidisciplinary approach including surgery, chemotherapy, and radiation therapy has been employed. The rarity of cases in even major tumor centers has traditionally limited the ability to study this disease. Several novel pharmacologic and biologic treatment approaches are actively being developed, although long-term follow-up is needed for their substantiation. PMID- 16311112 TI - Massive allograft use in orthopedic oncology. AB - Allograft transplantation is a functional reconstructive option for large extremity osseous defects. Improvements in anatomic matching, infection prevention, allograft fixation, soft tissue reconstructions, and rehabilitation protocols have greatly influenced predictability and longevity of massive allografts. These demanding reconstructions require time, an experienced group of orthopedic surgeons working at an institution with access to a large volume of patients, and a reliable, modern bone bank to select the appropriate graft for each individual under safe conditions. This article discusses the current use of total condylar osteoarticular allografts, hemicondylar allografts, allograft arthrodesis, intercalary segmental allografts, hemicylindric intercalary allografts, and allograft-prosthesis composites. PMID- 16311113 TI - The use of prostheses in skeletally immature patients. AB - Prosthetic reconstruction in the skeletally immature patient is challenging because of the necessity to cope with expected disruption of limb growth after resection of one or more major growth plates and the high demands placed on implants by young patients. Although prosthetic reconstruction in children is fraught with many problems, it is usually possible to obtain a good functional limb at skeletal maturity. Extendable prostheses are readily acceptable to patients and their parents. The availability of noninvasive extendable prostheses is expected to lead to fewer complications, fewer surgical interventions, and even improved function. This article reviews the indication, evolution, and clinical and functional results of extendable endoprostheses, drawing from experience in the last 30 years of using such prostheses in children. PMID- 16311114 TI - Pelvic reconstruction techniques. AB - Partial resection of the pelvis or sacrum is an uncommon procedure, typically performed in the setting of tumors, severe infections, or trauma. The resultant defects, depending on the size and location, may cause significant postoperative morbidity or functional impairment. It is therefore essential that the surgeon be aware of all reconstructive options available and implement the most appropriate option for each individual patient. The purpose of this article is to review the functional consequences of the various pelvic resections and discuss the options available for reconstruction. PMID- 16311115 TI - Surgical management of metastatic spine tumors. AB - Recent advances in diagnostic tests and radiologic imaging, and the development of novel chemotherapeutic agents and radiation methods have greatly altered the treatment options in patients who have spinal tumors. Improvements in fundamental understanding of the mechanisms of bone metastases, developments in spinal instrumentation, and recent introduction of recombinant bone morphogenetic proteins for spinal reconstruction offer promising strategies in selected patients. Clear applications of the fundamental surgical oncology still apply to spinal tumors. This article considers recent advances in management of the metastatic tumors to the spine. PMID- 16311116 TI - Image-guided therapies in orthopedic oncology. AB - The treatment of patients who have malignancies of bone often require multidisciplinary care. The aim of this review is to outline percutaneous techniques that may be useful for the diagnosis and treatment of these patients. In particular, the existing procedures of percutaneous biopsy, alcoholization (ethanol ablation), vertebroplasty, kyphoplasty, osteoplasty, radiofrequency ablation, laser photocoagulation, and vascular embolization are reviewed. Aspects of each technique, including mechanism of action, patient selection, treatment technique, and recent patient outcome are presented. PMID- 16311117 TI - Polyomavirus disease in renal transplantation: review of pathological findings and diagnostic methods. AB - Up to 10% of renal transplant recipients can develop polyomavirus nephropathy (PVN) in the allograft, leading to premature graft failure. Recent studies have shown that early diagnosis of PVN before there is irreversible damage to the kidney can result in marked improvement of outcome with resolution of the infection in a large proportion of patients. Early histopathologic diagnosis is complicated by the subtle beginning of the infection and its multifocal nature. This review presents a comprehensive set of guidelines for the effective clinical use of urine cytology and quantitation of viruria and viremia in conjunction with the renal biopsy findings. The morphological features of PVN are presented with specific emphasis on the patterns of PVN that are based on the histological progression of the disease and that correlate with clinical outcome. Also discussed in the context of their clinical significance are the main virological and epidemiological aspects of the BK, JC, and SV40 polyomavirus infections. PMID- 16311118 TI - Polyoma virus infection of renal allografts: relationships of the distribution of viral infection, tubulointerstitial inflammation, and fibrosis suggesting viral interstitial nephritis in untreated disease. AB - Whether polyoma virus (PV) infection of renal allografts induces an antiviral or antigraft immune reaction is unclear. By examination of the relationships of tubular PV to graft inflammation and scarring, this study sought histological evidence of viral interstitial nephritis in allograft biopsies with untreated PV infection and compared the inflammatory indices to controls with acute rejection (AR). Morphological features including viral cytopathic changes (VCCs) and modified Banff 97 histological indices were evaluated in sections of 28 diagnostic biopsies from a group of patients receiving prednisone, tacrolimus, and mycophenolate mofetil at constant dosage before biopsy. Two-micrometer paraffin sections were stained for PV large T antigen (TAg) and for C4d, by immunohistochemistry. Tubular profiles with 1 or more nuclei expressing TAg per x200 field were scored using an interval scale (0-10; none to 91-100%) by 2 observers. Controls with AR (n = 38, TAg negative) were matched for time after transplantation and severity of Banff 97 interstitial inflammation (i) and tubulitis (t) scores. Median t scores for tubules with VCC or TAg or both exceeded scores for tubules without VCC or TAg (3 versus 0, P = .001). Tubular TAg score correlated with i score (r = 0.58, P < .01) and sum ct + ci score (r = 0.61, P < .001). Atrophic tubules in scars had persistent VCC and/or TAg. Interstitial plasma cells (75% versus 21%) and neutrophils (32% versus 0%) were more frequent, and interstitial fibrosis was more severe (ci >1 in 54% versus 21%) in polyoma virus nephropathy (PVN) than in the group with AR (P < .01). Intimal arteritis (0% versus 35.7%), peritubular capillary C4d (0% versus 47.4%), and interstitial hemorrhage (4% versus 37%) were almost exclusively found in AR (P < .01). Tubular inflammation in untreated PVN involves infected tubular profiles with greater severity than those without evidence of infection. The extent of tubular PV infection is proportional to interstitial inflammation and scarring. Tubulointerstitial inflammation in PV infection has significant qualitative differences from AR. Observations in these examples of untreated PVN suggest that the allograft inflammatory reaction may exhibit features of viral tubulointerstitial nephritis distinct from AR. PMID- 16311119 TI - Promoter hypermethylation profile of RASSF1A, FHIT, and sFRP1 in intracranial primitive neuroectodermal tumors. AB - Medulloblastomas (MBs) and supratentorial primitive neuroectodermal tumors (SPNETs) are histologically alike intracranial PNETs found in different anatomical locations of the brain. Current evidence suggests that hypermethylation of promoter CpG islands is a common epigenetic event in a variety of human cancers. The aim of this study was to investigate whether promoter hypermethylation of putative tumor suppressor genes was involved in both types of intracranial PNETs. We examined the methylation status at promoter regions of RASSF1A, FHIT, and sFRP1 by methylation-specific polymerase chain reaction in a cohort of 25 primary MBs, 9 primary SPNETs, and 3 MB and 2 SPNET cell lines. Our results revealed no promoter hypermethylation of RASSF1A, FHIT, and sFRP1 in 2 normal cerebellar and 5 normal cerebral tissue specimens examined. In contrast, promoter hypermethylation of RASSF1A was detected in 100% of primary MBs, 67% (6/9) of primary SPNETs, and all PNET cell lines. The frequency of promoter hypermethylation of RASSF1A was significantly lower in SPNETs than in MBs (Fisher exact test, P = .014). Treatment of RASSF1A-deficient PNET cell lines with 5-aza-2'deoxycytidine, a demethylating agent, restored RASSF1A expression, providing evidence that promoter hypermethylation contributes to transcriptional silencing. In addition, promoter hypermethylation of FHIT and sFRP1 was detected in 22% (2/9) and 11% (1/9) of SPNETs, respectively, but not in any MBs studied. In conclusion, our study demonstrates that promoter hypermethylation of RASSF1A is a common event in intracranial PNETs, whereas FHIT and sFRP1 are epigenetically affected in a fraction of SPNETs. PMID- 16311120 TI - Extramammary Paget's disease: analysis of growth signal pathway from the human epidermal growth factor receptor 2 protein. AB - Paget's disease is a skin cancer characterized by characteristic (Paget) cells scattered in the epidermis. Although its prognosis is generally favorable with surgical resection, the clinical outcome turns unfavorable in cases with recurrence and metastasis. Therefore, establishment of effective therapeutic regimens is required for advanced Paget's disease. The human epidermal growth factor receptor 2 (HER2) protein, a transmembrane growth factor receptor, is frequently overexpressed in malignancies, causing activation of the phosphatidylinositol 3 kinase (PI3K) and extracellular signal-regulated kinase (ERK) signal pathways. Recently, HER2-targeting molecular therapy using trastuzumab (Herceptin; Genentech, Inc, South San Francisco, Calif) was revealed to be effective in advanced breast cancers overexpressing HER2 protein. Here, we analyzed the correlation between activation of the HER2 signal pathways and clinicopathologic parameters of 36 extramammary Paget's disease samples from 34 Japanese patients, using immunohistochemical analyses for HER2, phosphorylated HER2, phosphorylated AKT, and phosphorylated ERK proteins. We found overexpression of the HER2 protein in 19.4% (7) of the lesions, 3 of which showed HER2 amplification by chromogenic in situ hybridization. Phosphorylated HER2 protein was detected in 12 lesions (33.3%), including 2 of the 7 HER2 overexpressing lesions. Phosphorylated AKT was detected in approximately 75.0% (27/36) and phosphorylated ERK in 38.9% (14/36). Both HER2 and AKT were simultaneously phosphorylated in 9 cases (25.0%) and HER2 and ERK in 9 cases (25.0%), but all 3 molecules were phosphorylated in only 1 sample. Phosphorylated ERK correlated with the maximum diameter of the tumors (P < .025), but other immunohistochemical parameters failed to show any correlation with clinicopathologic features. These results suggest the contribution of the HER2 signaling pathway to the pathogenesis and progression of some cases of extramammary Paget's disease, for which clinical use of molecular target therapy against the HER2 pathway is warranted. PMID- 16311121 TI - Expression of Aurora kinases A and B in normal, hyperplastic, and malignant human endometrium: Aurora B as a predictor for poor prognosis in endometrial carcinoma. AB - Aurora kinases such as Aurora A and Aurora B are key regulators of mitosis and have been reported to be overexpressed in various malignancies. However, the expression and localization of Aurora kinases in normal and neoplastic endometrial tissues remain undetermined. In the present study, immunohistochemical expression of Aurora A and B was examined in 40 normal, 30 hyperplastic, and 73 malignant endometria. The data were compared with the expression of Ki-67 and patient survivals. The expression of Aurora A and B at protein and messenger RNA levels was also examined using Western blotting and the reverse transcriptase polymerase chain reaction. The expression of Aurora A in normal endometrium was observed mainly in the proliferative phase and was decreased in the secretory phase. The Aurora A expression was significantly increased in carcinomas compared with normal proliferative endometrium; however, there was no correlation of Aurora A expression with Ki-67 expression or patient survival. The expression of Aurora B in normal endometrium was significantly higher in the proliferative phase than in the secretory phase. In endometrial carcinomas, the expression of Aurora B was correlated with Ki-67 expression and was significantly increased in high-grade tumors. In addition, patients with Aurora B-positive carcinoma showed poor prognosis compared with those with Aurora B-negative carcinoma (P = .0135). Accordingly, the present study indicates the aberrant expression of Aurora A and Aurora B in endometrial carcinomas and the clinical importance of Aurora B expression in relationship to patient prognosis. PMID- 16311122 TI - Sperm protein 17 expression defines 2 subsets of primary esthesioneuroblastoma. AB - Esthesioneuroblastomas (ENBs) are rare malignant tumors of the nasal vault, the origin, diagnosis, and management of which are still subjects of discussion. That there is no related prognostic factor or generally recognized therapeutic regimen highlights the need for further analyses of its underlying biologic features and investigations of new marker proteins that allow more reliable clinical testing. We here show that sperm protein 17 (Sp17) is expressed in the ciliated cells of the normal olfactory epithelium and in a proportion of primary ENB lesions. We found an association between Sp17 expression and metastases at relapse (P = .035), chromogranin expression (P = .014), and a female sex prevalence. A statistically nonsignificant relation was found between Sp17 and S-100, synaptophysin, and neurofilament expression. No correlation was also found between Sp17 expression and the proliferative capacity of the lesion that was evaluated by Ki-67 immunohistochemistry. The results of this study show the usefulness of Sp17 as a means of discriminating 2 subsets of primary ENB lesions and seem to suggest the existence of 2 distinct cell pathways in their origin and development. PMID- 16311123 TI - Nuclear translocations of beta-catenin and TCF4 in gastric cancers correlate with lymph node metastasis but probably not with CD44 expression. AB - Interaction of nuclear beta-catenin and TCF4 is the end point of canonical Wnt signaling, which is believed to trigger the transcription of multiple cancer associated genes, including CD44. So far, the combined status of beta-catenin and TCF4 and its relevance for lymph node metastasis and CD44 expression have not been well studied in gastric cancers (GCs). To address these issues, we examined 31 GCs, 17 premalignant tissues, 10 noncancerous gastric mucosae, 17 regional lymph node metastases, and 4 human GC cell lines (MGC803, MGC823, AGS, and HGC 27) using immunohistochemical and immunofluorescence staining, reverse transcriptase polymerase chain reaction, and Western blot analysis. Frequent TCF4 up-regulation and nuclear translocation of beta-catenin were found in both primary and metastatic tumors. Standard CD44 was detected in all gastric tissue samples. The frequency of variant CD44 expression increased in parallel with stepwise gastrocarcinogenesis and tumor spread, but the rates of detection did not match that of nuclear beta-catenin and TCF4, especially in the premalignant and noncancerous samples. The data from the 4 cell lines were in accordance with the in vivo findings in terms of beta-catenin nuclear translocation, TCF4 activation, and CD44 expression. Our results suggest an established Wnt signaling pathway in most GCs, a close correlation of beta-catenin/TCF4-mediated signaling with tumor dissemination, and the unlikelihood of a direct effect of activated Wnt signaling on CD44 expression. The influence of beta-catenin-TCF4 interaction on alternative CD44 splicing was not established. These 3 alterations may be regarded as unfavorable features of GC. PMID- 16311124 TI - Correlation of spectral imaging and visual grading for the quantification of thymidylate synthase protein expression in rectal cancer. AB - Quantification of protein expression in tissue sections stained by immunohistochemistry has traditionally involved visual grading techniques. However, if these results are to be used to predict tumor behavior and permit targeted therapy, there is a need for more accurate, objective, and reproducible methods. This study investigated the utility of spectral imaging as a method of quantifying thymidylate synthase protein expression in immunohistochemically stained sections of primary rectal cancer and normal rectal mucosa by comparing it with the current gold standard of manual visual grading. There was good correlation between estimates of thymidylate synthase stain intensity and area derived by spectral imaging and visual grading in both tumor and normal mucosal sections, suggesting that spectral imaging is a valid way of quantifying biologic sections stained by immunohistochemistry. PMID- 16311125 TI - Image-guided biopsy in the evaluation of renal mass lesions in contemporary urological practice: indications, adequacy, clinical impact, and limitations of the pathological diagnosis. AB - With increasing sophistication of imaging modalities, many small indeterminate renal masses are detected, posing therapeutic dilemmas. Minimally invasive techniques such as radio frequency ablation (RFA) and cryotherapy are increasingly used in such settings, making it essential to classify renal mass in biopsies. We evaluated indications, adequacy, spectrum, and limitations of pathological diagnoses in 52 (79%) of 66 adequate biopsies that impacted management by having conservative therapy (less than total nephrectomy). Indication for biopsies in this group was exclusively for indeterminate mass. Biopsies were categorized as 52% clear cell renal cell carcinoma (RCC); 11% papillary RCC; 23% oncocytic neoplasms, subdivided as oncocytoma (10), chromophobe RCC (1), and cannot rule out RCC (1); 8% spindle cell neoplasms; 2% round blue cell tumors; and 4% inflammatory. After biopsy, 29% underwent nephron sparing surgery, 36% underwent RFA, and 35% were followed up with observation only. One (2%) unresolved oncocytic neoplasm proved to be a chromophobe RCC in the total nephrectomy. Of the 19 patients, 15 who underwent RFA after diagnostic biopsy subsequently underwent post-RFA biopsy to assess therapy response. Complete ablation at first attempt was achieved in 12 of 15 patients. In contemporary urological practice, biopsies are increasingly performed for incidentally detected renal mass to influence the clinical management. Most biopsies can be classified into clinically relevant categories; however, caution is advised while interpreting oncocytic neoplasms because sampling and tumor heterogeneity may adversely affect interpretation. PMID- 16311126 TI - Serous papillary carcinoma of the endometrium arising from endometrial polyps: a clinical, histological, and immunohistochemical study of 13 cases. AB - Serous papillary carcinoma is an aggressive tumor. Point mutations in the p53 suppressor gene might explain in part the rapid growth of this malignant tumor and its unfavorable outcome. The aims of this study were to evaluate the behavior of serous papillary carcinoma developing in endometrial polyps and to assess the p53 protein overexpression. Patients included in this study were treated in our institution between 1982 and 2003. All clinical and pathological materials were examined. A p53 protein immunohistochemical analysis was performed on paraffin embedded tissues. Thirteen serous papillary carcinomas arising from benign polyps of the endometrium were identified. The patients' age averaged 73 years. All patients were treated surgically. After an average follow-up of 22 months, 54% of the patients were dead or alive with disease. Of 10 serous papillary carcinomas, 8 (80%) for which paraffin blocks were available overexpressed the p53 protein. A serous papillary carcinoma arising from benign polyps of the endometrium remains a malignant neoplasia with an unfavorable outcome even if the primary tumor is limited to the polyp. The high rate of protein p53 overexpression suggests that a p53 gene mutation occurs early in the disease and might explain the rapid growth of the tumor. PMID- 16311127 TI - Breast cancer in an MSH2 gene mutation carrier. AB - A 49-year-old woman presented with breast cancer. She is a member of a family with the hereditary nonpolyposis colorectal cancer syndrome for which a 2-base pair deletion in exon 11 of the mismatch repair gene MSH2 (c1705_1706 delGA) had been identified. Breast cancer is rare in the hereditary nonpolyposis colorectal cancer syndrome. Microsatellite analysis of the tumor showed a microsatellite instable pattern for markers Bat25, Bat26, and Bat40, and no changes for markers D2S123 and D5S346, a so-called microsatellite instability-high pattern. Immunohistochemical staining for the mismatch repair enzymes MSH2 and MSH6 was negative, whereas the tumor cells were positive for MLH1, a pattern suggestive for biallelic MSH2 gene inactivation. We tested the tumor for loss of heterozygosity of the MSH2 gene and found loss of the wild-type MSH2 allele. These data strongly suggest that the MSH2 gene was involved in the development of this breast tumor. PMID- 16311128 TI - Challenges in QOL assessment. Overview and introduction. PMID- 16311129 TI - Optimal timing for QOL assessments. PMID- 16311130 TI - Combining information across symptom studies. PMID- 16311131 TI - Presenting longitudinal data. PMID- 16311132 TI - Incorporating clinical significance into a study. PMID- 16311133 TI - Handling missing data. PMID- 16311134 TI - Quality of life from a patient's perspective: can we believe the patient? PMID- 16311135 TI - The impact of quality of life measurements on the patient. PMID- 16311136 TI - Future directions in QOL research. PMID- 16311137 TI - Au revoir, Auf Wiederzhen...... PMID- 16311138 TI - Sympathetic skin response in idiopathic and diabetic carpal tunnel syndrome. AB - BACKGROUND: In carpal tunnel syndrome (CTS), certain changes were expected in sympathetic skin response (SSR) because median nerve carries postganglionic unmyelinated fibres. PURPOSE: To investigate the median and ulnar SSR in idiopathic and diabetic CTS without autonomic dysfunction in hands and to find possible relations with electrophysiological features of these diseases. PATIENTS AND METHODS: SSRs were elicited by electrical stimulation on the supraorbital nerve and recorded from the median and ulnar territories in the hand from 20 diabetic patients with only CTS (29 hands), 24 idiopathic CTS patients (42 hands) and 13 normal subjects (26 hands). Hands with ulnar neuropathy at the wrist without symptoms and normal hands of unilateral CTS were excluded. In addition to classical parameters and comparative methods, SSR waveform changes and percentile method was used in finding abnormality. RESULTS: Median SSRs had significant delayed latency compared to ulnar latency in both CTS patients but this was not important clinically (1.38/1.37 s for idiopatic CTS; 1.43/1.36 s for diabetic CTS). Median and ulnar SSR amplitude, area, median/ulnar latency difference, amplitude and area ratio were compared and only median/ulnar latency difference and median/ulnar latency ratio were found different between the three groups. Four idiopathic CTS hands were outside of the limits or absent (9.5%). SSR waveforms were significantly different from normal subjects in CTS patients. P type SSR replaced M type in idiopathic CTS and N type in diabetic CTS. CONCLUSIONS: Findings regarding SSR parameters suggest that unmyelinated C fibers were affected in CTS. These values were not useful because they were too small. Data on SSR were not normally distributed and the percentile method seems to be more convenient for finding any abnormality in clinical practice. Also, SSR waveform analysis could give us valuable data and should add to the SSR examination parameters. PMID- 16311139 TI - "Dementia unmasked": atypical, acute aphasic, presentations of neurodegenerative dementing disease. AB - Acute onset of aphasia is most commonly due to a cerebrovascular event in the territory of the dominant hemisphere middle cerebral artery. An isolated and slowly progressive aphasia may occasionally be the presenting feature of a dementia syndrome. The longitudinal clinical, neuropsychological and neuroimaging findings of two patients with an acute, perioperative, onset of aphasia are reported, to show that acute aphasia may on occasion be the presenting feature of neurodegenerative dementing disorders, notwithstanding clinical diagnostic exclusion criteria for these conditions. PMID- 16311140 TI - Computed tomographic changes in lacunar syndromes. AB - BACKGROUND: Lacunes are small deep infarcts due to occlusion of long-perforating arterial branches. The clinical presentation and outcome are variable. The present study investigates the prognostic value of comparing repeated computed tomographic (CT) scans in patients with a lacunar syndrome (LACS). PATIENTS AND METHODS: From a series of 220 patients, subsequently admitted with a first-ever stroke, 32 were diagnosed as LACS. CT scans without contrast enhancement were performed on day 3 and 10 after onset of the symptoms. A lacune was considered as responsible for the LACS when its location corresponded to the expected cerebral hemisphere and when it changed in size and/or in X-ray attenuation on the CT scans from day 3 to 10. The size of the lacune was also measured and compared on both time points. The patients were divided into two groups according to the modified Rankin (R) scale at 3 months, in those who were independent (R 0-1-2) and in those who remained handicapped (R 3-4-5). RESULTS: The patients who remained disabled at 3 months were older and more severely impaired on admission, and had more frequent cardiac problems and cognitive decline than the independent ones. On CT scans of the brain a higher incidence of leukoaraiosis was observed in this group. Only in three patients of each group no relevant lacune could be observed both on day 3 and 10. The average size of the symptomatic lacune on day 3 and 10 was not statistically different between both groups, when comparing all patients with a LACS. When comparing only those patients with a visible symptomatic lacunar infarct on one of the CT scans, however, the average size of the symptomatic deep infarct was smaller in the independent than the handicapped patients on day 3 as well as on day 10. The lacune decreased in average size from day 3 to 10 in the former group but remained unchanged in the group of disabled patients. CONCLUSION: The outcome of patients with a LACS depends on several factors including age, cognitive and cardiac status, the presence of leukoaraiosis and also the size of the symptomatic lacune. PMID- 16311141 TI - Simple clinical predictors of brain lesions in patients with impaired consciousness: a cross sectional study from a rural, tertiary hospital in central India. AB - OBJECTIVE: To evaluate the diagnostic accuracy of vital signs for detecting brain lesions in patients with impaired consciousness in a rural setting. METHODS: We enrolled patients older than 12 years who presented with impaired consciousness of non-traumatic origin to the intensive care unit of a rural teaching hospital. The design was a cross sectional analysis of a hospital-based case series, independently comparing vital signs on admission (temperature, pulse, systolic and diastolic blood pressure) against a reference standard (final diagnosis). Diagnostic accuracy was measured by computing multi-level likelihood ratios, and area under the receiver operating characteristic (ROC) curve. RESULTS: We studied 386 patients of whom 242 (62.7%) were men. A total of 178 patients (46%) had a brain lesion. None of the clinical predictors could accurately distinguish between those with and without a brain lesion. The area under the ROC curve for pulse was 0.61 (S.E. 0.02); that for the systolic and diastolic blood pressure 0.70 (S.E. 0.02) each. Systolic BP provided informative test results in 29.7%, diastolic BP in 37.2% and pulse rate in 19.9% patients. CONCLUSION: Our findings suggest that the vital signs lack accuracy for ruling in or ruling out brain lesion in patients with impaired consciousness. PMID- 16311142 TI - Olanzapine versus propranolol in essential tremor. AB - PURPOSE: In this study, we evaluated the anti-tremor effect of olanzapine in patients with essential tremor (ET). MATERIAL AND METHODS: Thirty-eight patients with ET affecting upper limbs underwent a randomized, crossover study with olanzapine (20 mg/day) and propranolol (120 mg/day). Evaluation (baseline and 1 month after each drug) was done by using the tremor clinical rating scale and a self-reported disability scale. RESULTS: We did not find statistical difference in all tremor parameters before treatment in two groups. Both drugs improved essential tremor parameters. However, olanzapine significantly improved all tremor parameters except hygiene as compared to propronalol. CONCLUSION: These results suggest that olanzapine might be efficacious for essential tremor. Further clinical trials are indicated to establish the efficacy of olanzapine in patients with ET. PMID- 16311143 TI - Clinical and neuroradiological predictors of mortality in patients with primary pontine hemorrhage. AB - BACKGROUND AND PURPOSE: Primary pontine hemorrhage (PPH) accounts approximately for about 5-10% of intracranial hemorrhages, and PPHs are known to have a much less uniform prognosis. We aimed to evaluate the clinical and radiological predictors affecting the mortality in 32 patients with PPH. MATERIAL AND METHODS: We retrospectively evaluated the data of 32 patients with PPH admitted to our clinic between 1994 and 2004. We divided the patients into two groups: (1) patients who survived (14 patients), and (2) patients who died (18 patients). The two groups were compared for age, gender, diabetes mellitus, hypertension, initial clinical status, initial GCS, pupillary abnormalities, ophthalmoparesis, volume and localisation of hemorrhage, intraventricular and extrapontine extension, necessity of mechanical ventilation and hydrocephalus. The hematoma volumes were measured with the formulation described by Broderick. RESULTS: Eighteen patients (56%) died and 14 patients (44%) survived. The patients who died (61.3 +/- 8.8) were older than the survivors (56.4 +/- 11.0), but the difference was not statistically significant. The mean GCS was 4.4 +/- 0.2, the mean hematoma volume was 9.9 +/- 3.3 ml for patients who died and the mean GCS was 10.1 +/- 3.3, the mean hematoma volume was 3.3 +/- 1.2 ml for survivors (p < 0.001). Coma on admission (p = 0.001), extrapontine extension (p = 0.001), intraventricular extension (p = 0.019), necessity of mechanical ventilation (p = 0.007), hydrocephalus (p = 0.024), massive and bilateral tegmental localisation (p = 0.006) were found statistically significant predictors for mortality with univariate comparison, and coma on admission (p = 0.038) was the only significant predictor with multivariate regression analysis. CONCLUSION: In patients with PPH, it is important to know the prognostic factors for mortality for planning the treatment protocol, and coma and bad clinical status on admission was found the only significant prognostic predictor for mortality with multivariate regression analysis. PMID- 16311144 TI - Accuracy of somatosensory evoked potentials in diagnosis of mild idiopathic carpal tunnel syndrome. AB - OBJECTIVE: Concerning prevalence of carpal tunnel syndrome (CTS) and the difficulties with electromyography (EMG) and nerve conduction studies (NCS), this study was designed to evaluate the power of somatosensory evoked potential (SSEP) in CTS diagnosis among Iranian patients. PATIENTS AND METHODS: SSEP was performed on 100 asymptomatic hands of 50 healthy participants (40 female, age range 38-59 years) and on 61 hands of 46 patients (39 female, age range 34-58 years). Mean difference between N(20) latency of the middle finger and the wrist (median nerve innervation) as well as N(20) latency of the third finger and the fifth finger (ulnar nerve innervation) were measured. Using receiver operating characteristic (ROC) curve analysis, the upper limits of these variables were defined as 6.0 and 1.5 ms, respectively. Higher amounts in either of these variables were considered as positive SSEP for diagnosis of CTS. Measures of accuracy for SSEP were measured getting clinical diagnosis by two separate neurologists as the reference standard. In the patients' group who underwent both techniques of SSEP and EMG NCS, kappa statistic as the agreement coefficient between two procedures was calculated. RESULTS: Sensitivity, specificity, and likelihood ratios for positive and negative results of SSEP in diagnosis of CTS were 70.4%, 91.0%, 7.83 and 0.32, respectively. Sensitivity of EMG-NCS in diagnosis of CTS was measured as 81.9%. Measure of agreement between two procedures (kappa) was calculated as 0.42. CONCLUSION: This study showed that positive results of SSEP might have a role in diagnosis of CTS. However, larger studies to demonstrate diagnostic power of SSEP in comparison with EMG-NCS seem necessary. PMID- 16311145 TI - A case of fatal progressive neuropathy. Delayed consequence of multiple bites of a non-venomous snake? AB - Neurotoxicity is a common feature of the acute syndrome of systemic snake envenomation. In this report a patient with delayed onset of neurological manifestations after multiple bites of a non-venomous snake is described. The neurological cascade consisted of segmental painful muscle spasms presented several days after the snake bites, which gradually subsided and within a month was followed by muscle weakness in the legs. The latter progressed to flaccid quadriplegia with additional respiratory and autonomic failure leading to death after total disease duration of 5.5 months. The possibility that this unique neurological syndrome of fatal progressive neuropathy could be caused by either tetanus or an immune reaction initiated by snakebites is discussed. PMID- 16311146 TI - Spontaneous brain rupture: a complication of untreated hydrocephalus. AB - A case of infantile hydrocephalus with secondary spontaneous brain rupture is reported. As far as the authors' knowledge, this is the first case of spontaneous brain and ventricular rupture secondary to high-pressure hydrocephalus. It is a case of infantile hydrocephalus occurring due to the expansility of an infantile skull, which is normally not seen these days, and is a fatal sequela of untreated hydrocephalus. This rare complication occurs with the rupture of thinnest part of the ependymal layer of the ventricle, cerebral tissue, meningeal membranes, bone and scalp. PMID- 16311147 TI - Alexia without agraphia following cerebral venous thrombosis associated with protein C and protein S deficiency. AB - A 26-year-old right handed female was admitted to hospital with right homonymous hemianopia associated with alexia without agraphia. Her cranial magnetic resonance imaging and magnetic resonance angiography revealed a left occipital venous infarction due to thrombosis of the left transverse, sigmoid sinuses and the left internal jugulary vein. The underlying conditions were protein C and protein S deficiency associated with the use of oral contraceptives. To our knowledge, alexia without agraphia has never been described due to a venous infarction associated with hereditary thrombophilia in the literature. PMID- 16311148 TI - Intractable epilepsy in Turner syndrome associated with bilateral perisylvian hypoplasia: one case report. AB - Turner syndrome (TS) is the most frequent sex abnormality in females, generally associated with a 45,X0 karyotype. Although neurological complications are frequently part of the clinical picture, serious brain abnormalities are quite rare in TS. Epilepsy in TS is not frequent and so far only few cases have been reported, usually associated with cortical dysplasias. We report a Turner patient showing severe neurological impairment, refractory epilepsy and MRI finding of bilateral perisylvian hypoplasia. The possible dysgenetic role of X-chromosome on cortical morphogenesis is also discussed. PMID- 16311149 TI - Transient global amnesia heralding basilar artery thrombosis. AB - The cause of transient global amnesia (TGA) remains controversial. Focal cerebral ischemia, seizure, venous congestion, and migraine have all been proposed as underlying mechanisms. We describe a patient presenting with typical TGA who two days later developed a posterior circulation stroke due to basilar artery occlusion. He was treated successfully with intra-arterial thrombolytic therapy. Shortly thereafter, he had recurrent basilar artery thrombosis and jugular vein thrombosis, and was found to have a mucinous adenocarcinoma believed to be causing a hypercoagulable state. We believe this case supports the hypothesis that TGA can on occasion be caused by cerebral ischemia. PMID- 16311150 TI - Sylvian fissure dermoid cyst with intratumoral hemorrhage: case report. AB - It is rare for a dermoid cyst to develop intratumoral hemorrhage. A 61-year-old woman had a sudden-onset left hemiparesis and slow response to verbal requests for one week when unenhanced computed tomography scanning revealed a mixed iso- and hypo-dense heterogeneous lesion in the right fronto-temporal area. T1 weighted magnetic resonance imaging (MRI) of the brain showed a mixed hyper- and hypo-intense tumor in the right fronto-temporal area. The tumor became hyperintense on T2-weighted MRI and was faintly enhanced at tumor periphery on T1 weighted MRI. The tumor was excised and pathological examination revealed a dermoid cyst with intratumoral hemorrhage. The post-operative course was complicated by hemorrhage in the tumor bed, which was evacuated immediately. The patient improved and could walk without support two weeks after the second operation. After 1 year of follow-up, she was well and without neurological deficits. To the best of our knowledge after a literature review, only two previous cases of dermoid cyst have featured intratumoral hemorrhage. PMID- 16311151 TI - Retroclival epidural hematoma secondary to a longitudinal clivus fracture. AB - Retroclival epidural hematomas (RCEDH) make part of posterior fossa epidural hematomas (PFEDH) and represent an extremely rare entity not always easily diagnosed with computerized tomography (CT) due to beam hardening artifacts. The authors present a case of a child-to our knowledge-the first reported until now featured the rare combination of a longitudinal clivus fracture associated with concomitant epidural hematoma treated conservatively with favorable outcome of the patient. PMID- 16311152 TI - Adult unilateral periventricular pseudocysts with ipsilateral headache. AB - OBJECTIVES: Unilateral periventricular pseudocysts (PVPC), manifesting as ipsilateral headache have been only rarely reported in adults. CASE REPORT: In a 48-year-old woman, right-sided, pulsating headache occurred in 1999. Headache exclusively extended over the right head and periorbitally; it was frequently associated with a feeling of cold, lasted maximally for 1 day, and occasionally went along with nausea and right-sided lacrimation. Since January 2003, headache occurred daily with varying intensity. For headache, she was regularly taking doxepin (25 mg). Ordinary analgesics and tryptanes were only of minimal effect. Since 2001, left-sided hemi-hypesthesia occurred. CT and MRI scans of the brain disclosed right-sided cysts in the white matter with a maximal diameter of 1cm, partially grouped and partially disseminated and bilateral calcifications of the basal ganglia. Since hypoxemia/ischemia, subependymal hemorrhage, CNS infection, developmental defect of the mantle layer, chromosomal disorder, Zellweger syndrome, carbon monoxide intoxication, trauma, or mitochondriopathy were not causative, the etiology of PVPC remained questionable. CONCLUSIONS: Rarely, unilateral PVPC become symptomatic in adulthood, manifesting as ipsilateral headache, contralateral hemi-hypesthesia, depression, collapses, and slight cognitive decline. Clinical progression of the disease is not necessarily related to the progression of the imaging findings. PVPC should be included in the differential diagnosis of unilateral headache. PMID- 16311153 TI - Bilateral carpal tunnel syndrome associated to familial Mediterranean fever. AB - A unique case of bilateral severe carpal tunnel syndrome due to familial Mediterranean fever is reported. The syndrome was diagnosed by clinical examination and electrophysiological studies. Bilateral transverse carpal ligaments were released and the biopsy specimens revealed systemic type A amyloidosis. Up to our knowledge, the co-existence of bilateral carpal tunnel syndrome and familial Mediterranean fever has not been reported previously in the literature. PMID- 16311154 TI - Guillain-Barre syndrome and hemophagocytic lymphohistiocytosis in a patient with severe chronic active Epstein-Barr virus infection syndrome. AB - Epstein-Barr virus (EBV) infection causes a wide range of neurologic and hematologic manifestations. We report a 72-year-old Japanese male patient with severe chronic active EBV infection syndrome (SCAEBV) who presented with Guillain Barre syndrome (GBS) and developed hemophagocytic lymphohistiocytosis (HLH) several months after the onset of GBS. He showed acute onset of distal muscle weakness, ophthalmoplegia and bulbar palsy. Results of nerve conduction study revealed acute motor-sensory axonal neuropathy (AMSAN). His serum was positive for anti-LM1 IgG and anti-GM1b IgM. Titers of antibodies to EBV-related antigens indicated chronic reactivated EBV infection. Treatment with IVIg resolved the acute ophthalmoplegia, but there was no notable improvement in the AMSAN and bulbar palsy despite repeated. Finally, he developed refractory HLH resulting in a fatal outcome. In the present patient, it seems that SCAEBV was associated with the development of GBS and fatal HLH via parainfectious autoimmunity and direct infectious immune mechanisms, respectively. PMID- 16311155 TI - Posttraumatic pneumocephalus-induced bilateral oculomotor nerve palsy. AB - Bilateral third nerve palsy often points to the involvement of its nucleus. Third nerve palsy as a result of posttraumatic nuclear involvement is an extremely rare condition. A 23-year-old man presented with a depressed skull fracture after acute head trauma and had Glasgow Coma Scale Score of 9. The diameters of the pupils were 6.5 and 7.5 mm and they were not reactive to light stimulation. There was bilateral ptosis. Computed tomography (CT) relieved bilateral perimesensephalic pneumocephalus. We suggested that bilateral oculomotor nerve paresis might develop in association with posttraumatic bilateral perimesensephalic pneumocephalus, which affected the nucleus of the third nerve. PMID- 16311156 TI - Endoscopic third ventriculostomy in the management of Chiari I malformation and syringomyelia associated with hydrocephalus. AB - BACKGROUND: Chiari malformation with syringomyelia is conventionally managed with foramen magnum decompression and duroplasty. In presence of hydrocephalus, a ventriculoperitoneal shunt insertion has been the initial preferred mode of treatment. METHODS: Two patients of Chiari I malformation with syringomyelia who had associated hydrocephalus were initially treated with endoscopic third ventriculostomy (ETV). RESULTS: At follow up, both the patients symptomatically improved. Repeat MRI studies revealed collapse of the syrinx cavity and reduction of the Chiari malformation. CONCLUSIONS: Chiari malformation and syrinx associated with hydrocephalus may be effectively managed with endoscopic third ventriculostomy. PMID- 16311157 TI - Concurrent cerebral and axillary artery occlusion: a possible source of cerebral embolization from peripheral artery thrombosis. AB - Intracranial embolization usually arises from heart, precerebral artery or aorta but rarely the peripheral artery as retrograde upstream of embolus should be overcome. We encountered a woman who experienced a transient right acronumbness followed by a sudden onset of left hemiplegia and conscious change concurrently. Cranial computerized tomography revealed a hemorrhagic infarct at right lentiform nucleus. Angiography disclosed right axillary artery occlusion at the proximal portion without collateral circulation. Cardioaortic survey and coagulation profile were normal. Thrombectomy was done and radial flow was corrected rapidly. Post-thrombectomic heparinization did not bring complication. Therefore, a distant embolism from peripheral artery at the upper limb should be alerted for cerebral ischemia. A lower density of embolus and no-flow condition in her occluded axillary artery may facilitate the upstreamed embolization. Heparinization, under closed monitor, is not contraindicated for preventing restenosis after thrombectomy in case of hemorrhagic transformation in brain. PMID- 16311158 TI - A case with cholinesterase inhibitor responsive asymmetric posterior cortical atrophy. AB - A 55-year-old right-handed woman presented initially with mild amnestic and depressive episodes but slowly developed progressive neurobehavioral symptoms, indicative of posterior cortical atrophy in ensuing years. A more detailed neurobehavioral test suggested predominant right temporo-parietal dysfunction with executive functional deficits. SPECT and MRI findings revealed right unilateral temporo-parietal involvement. Cholinesterase inhibitor administration led to amelioration of symptoms. We suggest that cases of posterior cortical atrophy or visual variant of Alzheimer's disease may be responsive to cholinesterase inhibitor therapy; however, the observation in a single case should be confirmed on larger populations in a clinical trial design with placebo control. PMID- 16311159 TI - Downbeat nystagmus following classical heat stroke. AB - Cerebellar syndrome following classical heat stroke is rare. We report a case of a 39-year-old man who presented with a cerebellar syndrome including downbeat nystagmus. MRI taken after 3 days of admission was normal. The downbeat nystagmus lasted for 6 days and than gradually disappeared. The ataxia and dysmetria improved gradually over 2 months. A brain CT scan performed 3 months later revealed no cerebellar atrophy. This case is unique as there were no predisposing factors (e.g. hypomagnesemia) and follow-up revealed complete reversibility of the ophthalmological abnormality. It reveals that the vestibulocerebellum is particularly vulnerable to thermal injury. PMID- 16311160 TI - Rebleeding from a primary brain tumor manifesting as intracerebral hemorrhage (CNN 04/077, revised version). AB - Symptomatic spontaneous intratumoral hemorrhage is a rare event in a patient with a brain tumor (BT). Although the treatment of choice in such a case is surgical removal of both the tumor and the hemorrhage, the optimal timing for surgical intervention has not been clearly established, particularly in those who present with minimal neurological deficits and a small hemorrhage volume. Two cases of primary BTs manifesting as an intracerebral hemorrhage (ICH) are described, in which rebleeding from the tumor occurred shortly after the initial hemorrhage. The patients each presented with the sudden onset of a headache and minimal neurological deficits, and the neuroradiological workup was consistent with a diagnosis of hemorrhagic BT. Each patient remained neurologically stable, and elective surgery had been planned within 7 days of their admission, but rebleeding occurred 5 and 6 days, respectively, after admission. A BT manifesting as an ICH may rebleed shortly after the initial bleeding, and should be treated on an emergency basis instead of an elective basis regardless of the patient's neurological status on admission or hematoma volume on the initial CT scans. PMID- 16311162 TI - Solar ultraviolet radiation: definitions and terminology. AB - In the rapidly developing field of photobiology as it relates to solar ultraviolet radiation (UVR), there is a need as never before to ensure that definitions and terminology are current, correct, and standard. This article provides a basic definition of UVR; a review of correct UVR radiometric symbols, units, and nomenclature; defines extraterrestrial and terrestrial solar UVR; and reviews the measurement of biologically effective dose of solar UVR in humans. PMID- 16311163 TI - An overview of ultraviolet radiation, sunscreens, and photo-induced dermatoses. AB - The incidence of sunlight-induced skin aging and skin cancers, particularly melanoma skin cancer, has been increasing in many parts of the world. Authorities are recommending primary prevention programs to reduce cutaneous photodamage and skin carcinogenesis. An integral component of these programs is the use of protective clothing and effective sunscreens. Most modern sunscreens have highly efficient absorption or reflecting capabilities throughout ultraviolet B, partly ultraviolet A, and in some instances infrared wavelengths. Over the last several years, more efficient sunscreening ingredients have been developed for improved skin protection. More recently, direct evidence has demonstrated the effectiveness of sunscreens in their ability to reduce the incidence of solar keratoses. This article reviews the protectiveness of sunscreens and assays that predict their levels of protection. PMID- 16311164 TI - Ultraviolet immunosuppression: mechanisms and consequences. AB - It is well recognized that exposure to solar radiation has several detrimental consequences, both acute and chronic. The suppression of immune functions remains one of the most intriguing phenomena brought about by ultraviolet (UV) radiation. This concept has challenged experts from various disciplines including dermatology, immunology, and photobiology. Although controversies exist regarding the mechanisms involved, the consensus is that UV immune suppression contributes significantly to the growth of cutaneous malignancies--both melanoma and nonmelanoma skin cancer. It is therefore a critical issue to be addressed in the context of developing and using sun protection strategies. PMID- 16311165 TI - Sunscreen use in photodermatoses. AB - Photodermatoses are relatively uncommon, but most dermatologists will have to diagnose and treat them at some point. This article highlights several photodermatoses, their common symptoms and possible. Unfortunately, sunscreens marketed in the United States do not presently contain agents that are effective against longer wavelengths. There are agents available in other countries that offer better protection. It is hypothesized that these products will soon be available in the United States. Until such time, clinicians should recommend agents with avobenzones, benzophenones, and the physical blockers titanium oxides. Two other agents, dihydroxyacetone and iron oxide, may also offer benefit in some of these patients. PMID- 16311166 TI - Human safety and efficacy of ultraviolet filters and sunscreen products. AB - Ultraviolet (UV) filters are the active ingredients in sunscreens. The concentration and combination of UV filters determine the efficacy of sunscreens as measured by sun protection factor. The safety of individual UV filters, and, more generally, sunscreen products, is a matter of a few related components: objective toxicologic evaluation, phototoxicologic potential, and human health consequences of using products that may reduce some but not all of the solar UV. Of 16 UV filters approved by the US Food and Drug Administration, 9 are used in different combinations in the most currently marketed sunscreens. Most of these compounds are considered safe and effective alone or in combination with other UV filters based on extensive toxicologic/phototoxicologic evaluations and market history. The benefits from proper use of sunscreens outweigh real or perceived human health concerns, establishing a favorable benefit-to-risk ratio. Future UV filters will require complete human safety evaluations alone and in combination with select benchmark ingredients. PMID- 16311167 TI - Sunscreen product formulation. AB - On the surface, sunscreen products are pretty simple. They consist of a delivery vehicle containing one or more sunscreen active ingredients. When applied to the skin, these sunscreen actives intercept solar ultraviolet (UV) rays before they can damage the underlying skin. However, while conceptually simple, a detailed analysis reveals that sunscreen formulations are quite complex, requiring careful selection of sunscreen active and vehicle components to control multiple performance and in-use parameters.Thus, to enable a better understanding and appreciation of sunscreen products, the typical steps in formulating a sunscreen product are described. Throughout this process, the key is to apply scientific principles together with a bit of formulation art to holistically create the best sunscreen product that meets the design targets--a sunscreen product that people will use regularly and properly. PMID- 16311168 TI - Ultraviolet A radiation: testing and labeling for sunscreen products. AB - Conceptually, sunscreen products are quite simple. The ultraviolet (UV) filters in these products reduce the "dose" of solar energy to which the skin is exposed. Underlying this empirical notion are many complexities including measures of product efficacy and how to communicate this to consumers. The sun protection factor (SPF) test is and should remain the singular in vivo method for evaluating sunscreens. Additionally, substrate spectrophotometric measure of absorbance/transmittance and the calculation of the summary statistic, such as the critical wavelength (ie, lambda(c)), should be used as a means of evaluating broad-spectrum (ie, UVA) protection. Ideally, the photoprotective efficacy of sunscreen products will be communicated to consumers as an SPF no greater than 50 and a single designation of "broad-spectrum" to indicate long-wavelength UVA protection. PMID- 16311169 TI - Water resistance and extended wear sunscreens. AB - The effectiveness of a sunscreen product can be reduced by sweating, friction, water immersion, or any other force that has the potential to remove the product from the surface of the skin. Substantivity (resistance to removal by water or sweat) along with durability (resistance to removal by wear) during physical activity are important aspects of sunscreen performance along with the fundamental protection provided against sunburn and ultraviolet-induced skin damage. Standards for measuring substantivity and other performance characteristics are available in regulations or guidelines around the world, but these regulations vary concerning the labeling and communication of product performance to consumers. Sunscreens alone should not be relied on to prevent all of the possible harmful effects of sun exposure. Variation between individuals, the substantivity and durability of products, and exposure conditions all play a role in the performance of sunscreens. PMID- 16311170 TI - Sunscreens and hair photoprotection. AB - Photoprotection as it pertains to hair is not a common topic addressed by the dermatologist. Hair is nonliving and requires no protection from ultraviolet (UV) radiation because carcinogenesis of the hair shaft itself is not possible. If hair proteins are altered by sun exposure, damaged hair can be removed and replaced by new growth. Thus, at first glance, the whole issue of photoprotection for the hair might seem irrelevant; however, patients frequently consult the dermatologist for advice on hair growth and appearance problems. Hair photoprotection is an important part of maintaining the cosmetic value of the hair shaft. This article focuses on the chemical effects of UV radiation on the hair shaft, hair photoaging, intrinsic hair UV photoprotective mechanisms, and the use of hair sunscreens. The whole science of hair and photoprotection is currently in its infancy and an area of focused research within the hair care product and salon industries. PMID- 16311171 TI - Garments as solar ultraviolet radiation screening materials. AB - Although wearing clothing to protect one's skin from the harmful rays of the sun is not new practice, this practice is of recent increasing interest. This article discusses 1) three types of protection (sunburn, precancerous skin lesion development, and photoaging) that can be realized by covering the skin with fabric, 2) the process by which some garments come to be labeled with information about ultraviolet radiation protection advantage, 3) the meaning of the information provided on product labels, 4) practical guidelines that can be used to decide which summertime garments having no stated sun protection information are the best for wearing out-of-doors, and 5) the pros and cons of using fabric and sunscreen lotions for sun protection. PMID- 16311172 TI - Compliance and sunscreens. AB - Part of the controversy surrounding sunscreens deals with issues of compliance. Sunscreens do not work if they remain in the bottle. It is estimated that the average United States adult uses less than one bottle of sunscreen per year. Clearly, this is indicative of poor compliance, since one bottle, if applied as directed on a daily basis, should last 1 month. What are some of the issues surrounding compliance? Why is compliance a problem? What can be done to improve sunscreen compliance? These are some of the issues addressed in this article. PMID- 16311173 TI - Novel emerging sunscreen technologies. AB - Because of increases in the number of skin cancers diagnosed annually, adverse effects of ultraviolet (UV) rays are being recognized, and major public education programs have been undertaken concerning photoprotection, including the use of sunscreen. In daily life, UV exposure is unavoidable; therefore sunscreen should be used regularly. Development in sunscreen manufacturing has grown tremendously in the last decade. Sunscreen active ingredients now are incorporated into cosmetics products to minimize photoaging changes. With the advances in technologies, many new UV filters have been developed recently. These have improved efficacy and safety. This article reviews these new filters, along with regulatory issues in the United States. PMID- 16311174 TI - Public health challenges in sun protection. AB - Sunscreens are a popular choice for protection from ultraviolet radiation, and hence, important components in the public health campaign to reduce the burden of skin cancer. Public health messages in skin cancer prevention have been used effectively in educational campaigns. The benefits of sunscreen extend beyond skin cancer prevention into other aspects of health and disease prevention: sunscreen decreases the risk for sunburn during physical activity outdoors and seems not to increase the risk for osteoporosis. Public health efforts have laid a solid foundation on which to face the continuing challenge of promoting and developing effective public health campaigns and health policies that encourage sunscreen use, sun protection, and the primary prevention of skin cancer. In this article, the controversies, concerns, and challenges of sunscreen use as it relates to public health are discussed. PMID- 16311176 TI - Informed consent to undergo treatment for substance abuse: a recommended approach. AB - With more than 3 million persons receiving substance abuse treatment per year in the United States and with increasing interest in treatment outcomes, there is a need for closer attention to all aspects of the treatment process. However, minimal attention has been given to informed consent as a way of enlisting client engagement and active participation in treatment. Although there is some literature on informed consent in substance abuse research, the literature on informed consent to undergo substance abuse treatment is very limited. Incorporating informed consent into substance abuse treatment is recommended as part of motivational interviewing. Standard treatment consent issues include (1) the clinical characteristics of the problem, including diagnosis; (2) treatment recommendations; (3) the risks and benefits of treatment; (4) the financial costs of the intervention; (5) alternative services or interventions should a client refuse the recommended form of care; and (6) freedom to choose or refuse treatment. This article provides a background for informed consent procedures to facilitate client engagement in substance abuse treatment and suggests needs for future research on informed consent to undergo substance abuse treatment. PMID- 16311177 TI - Use of a low-cost incentive intervention to improve counseling attendance among methadone-maintained patients. AB - Despite the importance of counseling in methadone maintenance treatment, many patients do not take advantage of these services. Incentives for attending group counseling were offered to methadone-maintained patients during an orientation phase of treatment or during required attendance at a relapse group later in treatment. Upon attending each counseling session, patients could draw for prizes under an escalating draw system with a 50% probability that draws would result in a prize. Incentives included small (1dollar), moderate (5 dollars), and large (20 dollars) prizes, with chances of winning inversely related to prize costs, and a maximum possible total of 160 dollars per patient. It was anticipated that this policy would provide a relatively low-cost approach to improving counseling attendance in our methadone clinic. The incentive policy significantly increased the percent of counseling sessions attended (52% vs. 76%) and promoted periods of continuous attendance. These data further support the effectiveness of low-cost incentive programs in enhancing counseling attendance among methadone patients. PMID- 16311178 TI - Patterns of heroin, cocaine, and alcohol abuse during long-term methadone maintenance treatment. AB - Individuals' use of heroin, cocaine, and alcohol during long-term methadone maintenance treatment (MMT) was studied. Prospectively collected data from 103 heroin-addicted individuals who were consecutively admitted for MMT and remained 2 years in treatment were evaluated. The patients were assessed every 6 months with a standardized interview. Three longitudinal patterns of drug abuse were identified. A proportion of patients abstained fully from their particular drug use (26% from heroin, 39% from cocaine, and 19% from alcohol); a proportion (39%, 32%, and 47%, respectively) switched between periods of abuse and nonuse of these drugs; and chronic drug users (34%, 28%, and 33%, respectively) continued use, including periods of daily abuse throughout MMT. Different therapeutic interventions may be needed in patients with different longitudinal patterns of additional substance use during MMT. PMID- 16311179 TI - Evidence-based treatment: why, what, where, when, and how? AB - Research and clinical perspectives are blended in this commentary on the rapidly emerging requirement for evidence-based treatment (EBT) in substance abuse programs. Although, historically, it has not been a standard of care in behavioral health, there are sound scientific, ethical, and compassionate reasons to learn and deliver an EBT as it becomes available. This article explores a series of issues, including the following: (1) Why should EBTs be used in substance abuse treatment? (2) What kinds of treatment are EBTs, and how are they determined? (3) Where can EBTs be implemented--at what levels of service delivery? (4) When should EBTs be used? and (5) How do clinicians learn EBTs? Potential pitfalls in implementing EBTs are also considered. PMID- 16311180 TI - "Painting a picture of the client": implementing the Addiction Severity Index in community treatment programs. AB - This article discusses the implementation of the Drug Evaluation Network System (DENS) version of the Addiction Severity Index (ASI) among substance abuse treatment providers in Los Angeles County. Thirty-two trained and certified treatment professionals from 14 programs participated in focus group interviews about their use of and experiences with the ASI. All 14 programs were currently administering the ASI at the time of the study. Qualitative interviews revealed variation among the programs in terms of when staff administered the ASI and how they used it in clinical care. Although the ASI creates a comprehensive picture of substance-abusing patients and their functioning, the timing of its administration seems to significantly affect the usefulness of the information. The findings illustrate the importance of practicality, flexibility, and ongoing training for the successful implementation of evidence-based practices. PMID- 16311181 TI - Effectiveness of workshop training for psychosocial addiction treatments: a systematic review. AB - Workshop training for psychosocial substance abuse treatment has been an important part of the transfer of evidence-based approaches into larger practice. Although they are widely used, training methods such as self-study reading, internet-based courses, and educational workshops have not traditionally been the focus of empirical investigations. Based on electronic and manual searches of the literature, we summarize 17 evaluations of workshop training that describe the training program and the educational outcomes. In general, training tends to improve attendees' knowledge, attitudes, and confidence in working with clients who have substance abuse problems. Some skill improvements, when measured, are usually seen immediately after training but are less often maintained over a longer time. Extended contact, through follow-up consultation, supervision, or feedback, appears to be necessary for the long-term adoption of skills. There are also a number of institutional factors that may influence the extent to which providers adopt new practices. Given the popularity of this training format, the role of workshop training needs to be a focus of future evaluative research. PMID- 16311182 TI - Methamphetamine treatment: trends and predictors of retention and completion in a large state treatment system (1992-2002). AB - This report describes trends in treatment admissions for methamphetamine/amphetamine (MA) abuse from 1992 to 2002 in California and assesses predictors of treatment retention and completion. Results show such admissions increasing fivefold and representing a growing proportion of overall treatment admissions. Patients admitted for MA abuse were increasingly diverse in race/ethnicity, older in age, and more frequently under legal supervision status over time. There was a decrease in injection drug use. Several user characteristics played consistent roles as risk factors for noncompletion and shorter treatment retention for both residential and outpatient admissions: having lower than a high school education, being younger at treatment admission, having a disability, having greater severity of MA use, and using injection drugs. Consistently, those with legal supervision status at admission had higher completion rates and longer retention than those reporting no legal status. Overall, findings suggested that clients with greater socioeconomic disadvantage and more severe problems may require greater efforts (e.g., services) to be retained in treatment. PMID- 16311183 TI - Buprenorphine tablet versus liquid: a clinical trial comparing plasma levels, efficacy, and symptoms. AB - We evaluated peak plasma concentrations, trough concentrations, and the 24-hour area under the concentration curve (AUC(0-24 h)) during maintenance with sublingual (SL) liquid or tablet formulations in 57 opiate-dependent volunteers. Study participants were assigned randomly to one of three SL daily buprenorphine dose pairs and maintained for 2 weeks with the liquid formulation followed by 2 weeks with the corresponding tablet dose. Plasma samples were obtained after at least 10 days of maintenance with the liquid formulation and after at least 10 days of that with the tablet formulation. The bioequivalence of the tablet compared with the liquid doses ranged from 57% to 75% based on peak concentrations, from 102% to 108% based on trough concentrations, and from 66% to 86% based on 24-hour AUC, but there was a large intersubject and intrasubject variability in plasma concentrations, with greater variability following tablets than liquid. Measures of withdrawal symptoms or illicit opioid use were not associated with buprenorphine dose, formulation, or plasma buprenorphine levels. PMID- 16311184 TI - Resource use of elderly emergency department patients with alcohol-related diagnoses. AB - This study evaluates resource use associated with alcohol-related admissions through the emergency department (ED) by older adults. Data from 11 states were extracted from the Healthcare Cost and Utilization Project State Inpatient Databases. The study results revealed that the presence of a secondary alcohol related diagnosis significantly increased resource use (37-119% for length of stay and 126-343% for charges; p < .05) associated with the top 10 International Classification of Diseases, Ninth Revision, Clinical Modification Clinical Classifications Software diagnoses. They also showed that admissions with an alcohol-related primary diagnosis had lower associated charges (2,172 dollars; p < .05) and longer lengths of stay (0.3 days; p < .05) than other types of ED admissions. Proper linkages to substance abuse treatment services should be instituted and coupled with medical treatment to limit the additional resource use burden of alcohol-related admissions. PMID- 16311185 TI - United Kingdom substance misuse treatment workers' attitudes toward 12-step self help groups. AB - Research has highlighted the benefits of professional substance misuse treatment workers facilitating their clients' involvement in 12-step self-help groups such as Alcoholics Anonymous (AA) and Narcotics Anonymous (NA). This study used a cross-sectional survey design to explore the attitudes of substance misuse treatment staff working in the English West Midlands region toward AA and NA and the 12-step philosophy. A total of 346 treatment workers responded (71%). Fewer than 10% used the 12-step model in their treatment work, and only a third felt that their clients were generally suited to AA or NA. Fewer than half (46%) said that they were likely to recommend that their clients attend a self-help group meeting. Staff with nursing qualifications were more likely to have a positive attitude toward AA/NA than their non-nursing colleagues, and there was a statistically significant association between the level of reported spirituality of the workers and the likelihood of them recommending to clients that they attend AA or NA meetings. These results highlight differences in the role of AA/NA within the substance misuse treatment system in the United Kingdom and that in the United States; possible explanations are discussed. PMID- 16311186 TI - Impact of drug treatment on subsequent sexual risk behavior in a multisite cohort of drug-using women: a report from the Women's Interagency HIV Study. AB - BACKGROUND: The evidence that drug treatment programs are associated with changes in sexual behavior and, thus, have prevention benefits beyond addiction is inconclusive. We examined whether entry into drug treatment was associated with subsequent alterations in sexual behavior among a group of drug-using women. METHODS: Data were collected semiannually via structured interviews over 8 years. Generalized estimating equations evaluated the relationship between self-reported drug treatment at each visit and sexual abstinence and consistent condom use in the subsequent 6-month period. RESULTS: In this sample (N = 1,658; mean age, 37.3 years; 57.5% African American; 80.3% HIV positive; 49.6% crack/cocaine users), 40% reported being in a variety of drug treatment programs. Those undergoing drug treatment (vs. those not) were less likely to become sexually active (adjusted odds ratio [AOR], 0.83; 95% confidence interval [CI], 0.76-0.91); this association was unchanged when the frequency of attendance and number of different drug treatment programs were evaluated. Drug treatment was not associated with subsequent consistent condom, regardless of frequency of attendance, but involvement in at least three treatment programs was (AOR, 1.40; 95% CI, 1.00-1.97). CONCLUSIONS: Additional efforts are needed to integrate effective sexual risk reduction programs into drug treatment settings; expanding access to different types of drug treatment modalities may be indicated. PMID- 16311187 TI - Identification of signatures in biomedical spectra using domain knowledge. AB - OBJECTIVE: Demonstrate that incorporating domain knowledge into feature selection methods helps identify interpretable features with predictive capability comparable to a state-of-the-art classifier. METHODS: Two feature selection methods, one using a genetic algorithm (GA) the other a L(1)-norm support vector machine (SVM), were investigated on three real-world biomedical magnetic resonance (MR) spectral datasets of increasing difficulty. Consensus sets of the feature sets obtained by the two methods were also assessed. RESULTS AND CONCLUSIONS: Features identified independently by the two methods and by their consensus, determine class-discriminatory groups or individual features, whose predictive power compares favorably with that of a state-of-the-art classifier. Furthermore, the identified feature signatures form stable groupings at definite spectral positions, hence are readily interpretable. This is a useful and important practical result for generating hypothesis for the domain expert. PMID- 16311188 TI - Relationship of spasticity to knee angular velocity and motion during gait in cerebral palsy. AB - This study investigated the effects of spasticity in the hamstrings and quadriceps muscles on gait parameters including temporal spatial measures, knee position, excursion and angular velocity in 25 children with spastic diplegic cerebral palsy (CP) as compared to 17 age-matched peers. While subjects were instructed to relax, an isokinetic device alternately flexed and extended the left knee at one of the three constant velocities 30 degrees/s, 60 degrees/s and 120 degrees/s, while surface electromyography (EMG) electrodes over the biceps femoris and the rectus femoris recorded muscle activity. Patients then participated in 3D gait analysis at a self-selected speed. Results showed that, those with CP who exhibited heightened stretch responses (spasticity) in both muscles, had significantly slower knee angular velocities during the swing phase of gait as compared to those with and without CP who did not exhibit stretch responses at the joint and the tested speeds. The measured amount (torque) of the resistance to passive flexion or extension was not related to gait parameters in subjects with CP; however, the rate of change in resistance torque per unit angle change (stiffness) at the fastest test speed of 120 degrees/s showed weak to moderate relationships with knee angular velocity and motion during gait. For the subset of seven patients with CP who subsequently underwent a selective dorsal rhizotomy, knee angular extension and flexion velocity increased post operatively, suggesting some degree of causality between spasticity and movement speed. PMID- 16311189 TI - Implicit motor learning of a balancing task. AB - This study examined the implicit learning of a balancing task. Three treatment conditions were constructed using different motor learning strategies. In two of the treatment conditions, explicit learning of the balancing task was impeded by using either an analogy or an errorless learning technique. In the third treatment condition, participants learnt the task by discovery learning, which typically results in explicit knowledge. It was hypothesised that in the analogy and errorless learning conditions, learning of the balancing task would be implicit in character. Three criteria of implicit learning were used to test this hypothesis; the accumulation of few explicit rules, robustness under secondary task loading and durability over time. Although the discovery learners acquired more explicit rules, all groups appear to have acquired the skill implicitly, in that all groups were robust to imposition of a concurrent task load and over time. Indeed, balance performance with a concurrent verbal task was better than balance performance alone. Discussion focuses on the contribution of verbal and non-verbal processes to balancing. PMID- 16311190 TI - Postural strategy to keep balance on the seesaw. AB - This work investigates the kinematic and electromyography (EMG) strategy used by the central nervous system (CNS) to keep equilibrium during anterior-posterior balance on seesaws with different degrees of instability. The movement of hip, knee, and ankle were reconstructed using a 3D motion-analysis system and the EMG activities of selected ankle, knee, and hip muscles were recorded. Balance was kept mainly at the ankle joint. The EMG patterns of the gastrocnemius and anterior tibialis alternated between agonist and antagonist bursts. The agonist burst started before the end of the lengthening phase and was prolonged until the end of the shortening phase. The EMG activities of the muscles crossing the knee and hip joints were characterized by a pattern of generalized co-activation. The movements at these two joints were very small, suggesting a neural or biomechanical constraint underlying the operations of the equilibrium control. Our results also indicate that the strategy to keep balance on the seesaw is qualitatively the same for the different levels of mechanical demands in terms of the seesaw's instability. PMID- 16311191 TI - Cervical joint position sense: an intra- and inter-examiner reliability study. AB - OBJECTIVES: To evaluate the intra- and inter-examiner reliability of neck active joint position sense measurements in different head movements. METHODS: Participants had to reproduce actively a specific angle-target in all movements (flexion, right/left rotation, right/left side flexion) from two initial positions, sitting and standing by using a 3D ultrasound-based motion analysis device. Three tests were employed to assess intra-examiner reliability and two examiners used for the inter-examiner reliability. Absolute error (AE) and variable error (VE) indices were used to assess the repositioning accuracy. Intraclass correlation coefficient (I.C.C.(1,1)), standard error of measurement (S.E.M.), smallest detectable difference (S.D.D.) indices were calculated for the analysis of the results. RESULTS: Both AE and VE yielded poor to moderate I.C.C.s in any movement and position (-0.01 to 0.50 and 0.01-0.25, respectively). AE presented higher I.C.C.s estimates than the VE but the S.D.D.s were similar for both indices. Regarding the AE, the standing position yielded higher I.C.C. estimates (0.15-0.68) than the sitting position (-0.01 to 0.43) but the S.E.M. (1.2-3.0 degrees and 1.5-3.5 degrees, respectively) and S.D.D. values (123.3 191.8% and 139.9-203.8%, respectively) showed no specific trend in favour of any position. The VE reliability indices showed that standing position was more reliable with less error than sitting. The inter-examiner data showed similar results to the intra-examiner study. CONCLUSIONS: The statistical analysis of the present experiments showed that the method employed for measuring cervical joint position sense is unreliable. However, it needs further research to identify the discriminatory power of these tests or if they are clinically unacceptable. PMID- 16311192 TI - Effect of equinus foot placement and intrinsic muscle response on knee extension during stance. AB - Equinus gait, a common movement abnormality among individuals with stroke and cerebral palsy, is often associated with knee hyperextension during stance. Whether there exists a causal mechanism linking equinus foot placement with knee hyperextension remains unknown. To investigate the response of the musculoskeletal system to equinus foot placement, a forward dynamic simulation of normal walking was perturbed by augmenting ankle plantarflexion by 10 degrees at initial contact. The subsequent effect on knee extension was assessed when the muscle forces were allowed, or not allowed, to change in response to altered kinematics and intrinsic force-length-velocity properties. We found that an increase in ankle plantarflexion at initial contact without concomitant changes in muscle forces caused the knee to hyperextend. The intrinsic force-length velocity properties of muscle, particularly in gastrocnemius and vastus, diminished the effect of equinus posture alone, causing the abnormal knee extension to be less pronounced. We conclude that the effect of ankle position at initial contact on knee motion should be considered in the analysis of equinus gait. PMID- 16311193 TI - Induced acceleration contributions to locomotion dynamics are not physically well defined. AB - Induced acceleration analysis quantifies the contributions of individual moments and forces to the accelerations, reaction forces, and powers produced during a task. The analysis has been advocated in the assessment of muscle and joint moment function during locomotion. However, results and interpretations drawn from the analysis have differed considerably between studies. The purpose of this paper is to assess whether induced acceleration contributions to locomotion dynamics are physically well defined. The assessment was facilitated by the analyses of a simple, theoretical locomotor task using different models. The task was based on a planar, rigid-body simulation in which joint moments at the hip, knee, and ankle posturally supported the configuration of the body as it rolled forward in a pendular motion. Induced acceleration analyses were performed using four models that completely described the simulated dynamics of the task but represented progressively fewer degrees of freedom. The contributions of each joint moment to the ground reaction force and trunk and leg powers differed between models, even though the net contributions were identical for all models and consistent with the simulation. Moreover, when all body degrees of freedom were represented in the model, large power redistributions between the trunk and leg were attributed to individual joint moments. However, these redistributions mostly cancelled such that the net redistribution was modest. In a single-segment model, this net redistribution was attributed entirely to gravity, without any cancellation of power flows. To conclude, induced acceleration contributions to the dynamics of a task are not physically well defined. The application of the analysis in the assessment of muscle and joint moment function during locomotion should be critically reevaluated. PMID- 16311194 TI - Postural control in male patients with hip osteoarthritis. AB - Earlier studies have demonstrated that postural control is worse in patients with knee osteoarthritis (OA) than in control subjects, whereas little information is available about the postural control in patients with hip OA. The aim of this study was to investigate the standing balance in different test conditions in men with hip OA and to compare the results with those of age- and sex-matched healthy controls. Twenty-seven volunteers 47-64-year-old men with hip OA and 30 randomly selected, healthy age-matched men were tested using the sensory organisation test (SOT). The center point of force velocity (CPFV (cm/s)) was also determined during one- and two-footed standing. There was no difference between the patients and controls in the SOT test, in the strategy analysis or during one-footed standing. There were no significant differences in CPFV values between the better and worse hip side. Fatiguing exercise had no effect on two-footed CPFV with eyes open, but when the eyes were closed both two and one-footed CPFV values were significantly increased (p<0.01-0.05) in both groups. No differences were found in fatiguing exercise between control and OA subjects with eyes open or closed or in one-footed trials. In OA patient's subjective pain was related to increasing CPFV (p<0.05). In conclusion, the hip OA had no effect on static balance in men. PMID- 16311195 TI - Understanding falls in osteoporosis: the viscoelastic modeling perspective. AB - Unsteadiness and increased postural sway predispose individuals with osteoporosis to falling. We examined this relationship from a biomechanical perspective using a viscoelastic model. Specifically, we investigated the center-of-pressure (COP) in quiet stance in 37 postmenopausal women (aged 42-79 years), diagnosed with osteopenia or osteoporosis and who engaged in a range of activity levels. Subjects stood on a force platform for two 20s intervals: one with eyes open (EO) and the other with eyes closed (EC). The COP signals were used to compute traditional measures of sway: standard deviation, range, and mean velocity. Based on the difference between the COP and its smoothed representation, we used the "COP minus center-of-mass" signal to compute selected properties of postural mechanics (stiffness and viscosity) and dynamics (frequency and damping) of stance. Sway variability and incidence of falls were not correlated even though our subjects displayed higher sway than age-matched subjects from other studies. Bone density was correlated with frequency in the mediolateral (ML) plane with EO (r=0.50, p<0.002). Irrespective of age, the decrease in postural frequency and increase in postural viscosity in the ML plane were determinants of imbalance and accounted for 18% and 8% of the variance, respectively. With EC, an additional 10% of the variance was contributed by the ML range. The increased ML viscosity was the sole predictor of falls accounting for 20% of the variance of the tendency to fall. Thus, ML frequency plays a fundamental role in preserving balance in that a low value is an important predictor of falls. Finally, although being physical activity may reduce sway, physical activity fails to improve poor postural strategies. To conclude, increases in viscosity and range elucidate further the unique strategy of ML body sway in individuals with osteopenia or osteoporosis and provide insight into how posture may be maintained by those who fear falling. PMID- 16311196 TI - Mechanisms underlying age-related differences in ability to recover balance with the ankle strategy. AB - Falls cause substantial death and morbidity in the elderly. An important prerequisite to the development of fall prevention interventions is improved understanding of the biomechanical and neuromuscular variables that govern ability to recover balance. In the present study, we examined the relative importance of strength versus speed of response variables in explaining differences in balance recovery performance with the ankle strategy between young and elderly women. Twenty-five young (19-36 years) and 25 community-dwelling elderly women (66-90 years) participated in balance recovery experiments. Subjects were supported in an inclined standing position by a horizontal tether and instructed to recover an upright vertical standing position by contracting their ankle muscles. The maximum initial lean angle from which they could recover balance without release of the tether (which depends primarily on strength) was 19.6% smaller for elderly than young. The maximum initial lean angle from which they could recover balance after the tether was suddenly released (which depends on both strength and speed of response) was 36.1% smaller for elderly than young. Moreover, between-group differences in performance were related to both strength and speed of response. Peak ankle torque was 7.7% smaller in elderly than young during dynamic recovery trials; reaction time was 27% slower in elderly, due to a lengthened muscle response latency, and rate of ankle torque generation was 15.6% slower in elderly. These results suggest that differences in ability to recover balance between young and elderly women associate with variables related to strength and speed of response, and that exercise-based fall prevention programs should include balance and agility training, in addition to strength training. PMID- 16311197 TI - Comparisons of the joint moments between leading and trailing limb in young adults when stepping over obstacles. AB - Twenty young healthy adults walked and crossed obstacles of heights of 10%, 20% and 30% of their leg lengths and kinematics and kinetics of their gait analysed. Three-dimensional joint moments of the lower limb were calculated with inverse dynamics analysis. Peak moments and crossing moments (joint moments at the time when the swing foot is above the obstacle) were obtained. Most of the peak moments of the supporting limb joints did not show the same height effects as the corresponding crossing moment components. This suggests that the true effects of obstacle heights on the joint kinetics during obstacle crossing cannot be obtained solely through comparisons of peak moments. For inter-limb comparisons the peak and crossing moments were different, suggesting that the leading and trailing limb adopted different kinetic strategies during obstacle crossing. The findings may help to shed light on further investigation on the motor control strategies of the central nervous system for obstacle crossing. PMID- 16311198 TI - Validation of a visual gait assessment scale for children with hemiplegic cerebral palsy. AB - This study reports the experience of reliability testing and validation of a visual assessment of gait based on the Physician Rating Scale in children with hemiplegic cerebral palsy. Comparison with three-dimensional gait analysis showed excellent correlation between observers and full gait analysis for sagittal plane motion at the knee in stance. Inter and intra-rater reliability showed moderate to almost perfect agreement for foot contact characteristics and ankle in stance. Use of this scale may be helpful in assessment of children where frequent analysis of these characteristics is needed. PMID- 16311199 TI - Leg lengthening as part of gait improvement surgery in cerebral palsy: an evaluation using gait analysis. AB - Eleven children with hemiplegic or an asymmetric diplegic cerebral palsy who had a preoperative leg length discrepancy of more than 2.5 cm underwent gait improvement surgery which included leg length equalisation. Sagittal plane kinematics and kinetics before and about 3 years after surgery for the lengthened limb and contralateral limb were evaluated. Preoperatively the unaffected limb had excessive stance phase flexion at the hip and knee, and dorsal flexion at the ankle joint. These changes could be partially reversed to produce a kinematic gait pattern comparable to age matched normal controls on the uninvolved side after equalisation of leg length. PMID- 16311200 TI - A prospective study of gait related risk factors for exercise-related lower leg pain. AB - The purpose of this study was to determine prospectively gait related risk factors for exercise-related lower leg pain (ERLLP) in 400 physical education students. Static lower leg alignment was determined, and 3D gait kinematics combined with plantar pressure profiles were collected. After this evaluation, all sports injuries were registered by the same sports physician during the duration of the study. Forty six subjects developed ERLLP and 29 of them developed bilateral symptoms thus giving 75 symptomatic lower legs. Bilateral lower legs of 167 subjects who developed no injuries in the lower extremities served as controls. Cox regression analysis revealed that subjects who developed ERLLP had an altered running pattern before the injury compared to the controls and included (1) a significantly more central heel-strike, (2) a significantly increased pronation, accompanied with more pressure underneath the medial side of the foot, and (3) a significantly more lateral roll-off. These findings suggest that altered biomechanics play a role in the genesis of ERLLP and thus should be considered in prevention and rehabilitation. PMID- 16311201 TI - Partial weight bearing after surgery for fractures of the lower extremity--is it achievable? AB - Partial weight bearing is a generally accepted principle of rehabilitation following trauma or reconstructive surgery of the lower extremity. Individual dynamic loads during partial weight bearing to a given load level of 200 N were compared in 23 patients who had sustained a fracture of the lower extremity and 11 healthy volunteers using dynamic sole pressure measurements. Excessive dynamic loading compared with the statically pre-tested 200 N level was observed in all groups. Maximum force levels were up to 690 N in young patients and up to 580 N elderly patients beyond the prescribed static load. None of the healthy volunteers was able to keep within the given load of 200 N. The set load level was exceeded by at least 38 N (119%) in the elderly patient group. In comparison, elderly patients showed statistically significantly higher maximum forces than young patients during the first two test days (p=0.007 and 0.013). On the 3rd test day the maximum ground contact forces were on average 71 N higher than in the young patients group. Analysis of the force time integrals (impulses transferred to the ground) displayed higher values in the older again than in young patients. The differences were statistically significant during the first two test days (p=0.006 and 0.037). This study implies that the conventional concept of postoperative partial weight bearing starting from 200 N and a stepwise increase of the load level until full weight bearing is not valid during clinical practice. PMID- 16311202 TI - The kinematics and kinetics of turning: limb asymmetries associated with walking a circular path. AB - The biomechanics of changing direction while walking has been largely neglected despite its obvious relevancy to functional mobility. The world is filled with turns that must be negotiated. These turns carry an increased risk of injury due to a decrease in stability. A VICON 612 system measured joint kinematics and kinetics on 10 normal subjects for straight line walking (ST); turning, inside foot strike (IN); and turning, outside foot strike (OUT). All trials were completed at a self-selected walking speed and across a range of speeds from 0.6 to 1.3 m/s; the turn radius was 1 m. Significant differences between the conditions were detected using a mixed effects repeated measures ANCOVA with walking speed as a covariate. The most pronounced differences were demonstrated in the mediolateral ground reaction force impulse: in straight walking the impulses tended to shift the body toward the contralateral limb. In turning, the IN and OUT impulses shifted the body toward the ipsilateral and contralateral limbs, respectively. Knee flexion during stance was increased on the IN limb, while ankle plantarflexion increased on the OUT limb consistent with body lean during turning; differences in joint kinetics during turning were negligible. Self-selected turning was significantly slower than walking straight ahead (0.96+/-0.12 m/s versus 1.61+/-0.22 m/s) and turning at very slow speeds showed a non-uniform center of mass trajectory. Understanding the mechanisms of turning will provide insights driving design, therapy and intervention to increase functional navigation in amputees, the elderly and individuals with neuromuscular pathologies. PMID- 16311203 TI - A comparison of gastrocnemius muscle-tendon unit length during gait using anatomic, cadaveric and MRI models. AB - Computing the length of a specific muscle is often of interest both in the study of human movement and the optimization of surgical techniques. The objective of this study was to compare three widely used methods of calculating gastrocnemius muscle-tendon unit (MTU) length. Gastrocnemius MTU length was calculated from the sagittal knee and ankle kinematics of 20 normal adults using three different methods: one based on cadaveric dissection and joint manipulation, one based on anatomic computer software modeling with insertion points obtained from cadavers and the third based on MRI measurements. All methods were normalized to percent anatomic neutral length (%ANL) with 100% at full knee extension and neutral ankle plantar-dorsiflexion. At the longest length (terminal stance), mean values (% ANL+/-standard deviation) were found to be 99.3+/-0.8, 101.8+/-0.5 and 102.4+/ 0.7 for the MRI, cadaveric and anatomic methods, respectively. During the shortest length (early swing), the MRI, cadaveric and anatomic methods had mean values of 93.0+/-0.6, 92.2+/-2.0 and 91.9+/-2.1% ANL, respectively. All methods showed statistically significant differences (p<0.01) from each other in terminal stance and early swing, while loading and terminal swing lengths were similar among all methods. The gastrocnemius MTU calculated using the MRI-based method was found to be the shortest. Additional research is needed in order to validate a model for calculating gastrocnemius MTU length using in vivo methods of estimating the gastrocnemius moment arm at the knee and ankle. PMID- 16311204 TI - Editorial. PMID- 16311206 TI - Recording click-evoked myogenic potentials (CEMPs) with a setup for brainstem auditory evoked potentials (BAEPs). AB - AIM OF THE STUDY: Click-evoked myogenic potentials (CEMPs) originate in the sternocleidomastoid (SCM) muscle through a reflex loop involving the sacculus and the vestibular nerve. In this study we suggest that they can be picked up from the mastoid reference used for auditory evoked potential (AEP) recording by using standard filters for brainstem AEPs (BAEPs). They consist of a P13-N20 complex. METHODS: Fifty normal subjects were investigated. Recordings were performed with the same setting as that used for conventional BAEPs but without artifact rejection and using a wide time window (100 ms). Unilateral auditory stimulations were used. All acquisitions were performed in both sitting and supine positions. In nine subjects CEMPs and BAEPs were simultaneously recorded at both earlobe and both SCM muscles. RESULTS: In all subjects, a CEMP P13-N20 component could be evidenced in sitting, but not in supine position at both the ipsilateral earlobe and the ipsilateral SCM muscle. Its latency was 0.7 ms lower at the earlobe. It obeyed the same relationship to stimulus intensity at both earlobe and SCM muscle. CONCLUSION: These results demonstrate the possibility to get simultaneous information on the brainstem auditory pathways and on a reflex probably mediated through the sacculus and the vestibular nerve. Further studies on patients with selective vestibular nerve impairment should be conducted to confirm the clinical utility of this approach. PMID- 16311207 TI - Respiratory evoked potentials and occlusion elicited sympathetic skin response. AB - INTRODUCTION: Neurophysiological study of respiratory structures usually relies upon diaphragm electromyography and phrenic nerve conduction study, which do not assess the afferent sensory pathways. OBJECTIVE: To assess the feasibility of respiratory evoked potentials (REPs) and sympathetic skin responses (SSRs) elicited by inspiratory occlusion. METHODS: REPs and SSRs were studied in 12 healthy adults. REPs were elicited by inspiratory occlusions triggered by the physician within 1 s after the onset of a respiratory effort. They were recorded from C3, C4 and Cz needle electrodes (referenced to Fz). Each individual trial consisted of two superimposed 30-sweep averaged responses to inspiratory occlusions. SSRs were recorded from surface electrodes placed on the subject's hand and elicited by similar inspiratory occlusions. RESULTS: Reproducible REPs and SSRs were obtained in all subjects. Mean latencies of initial P1 and N1 cortical responses were 41 and 72 ms, respectively. SSRs were similar to those usually elicited by peripheral nerve electrical stimulation. CONCLUSION: Brief occlusion of inspiration induces cortical and sympathetic activation, both are easily recordable. Since REPs are considered to be the neurophysiological substrate of certain types of respiratory sensations and are altered in different chronic respiratory diseases, they, in addition to SSR, represent attractive new techniques that may provide better understanding of respiratory dysfunction. PMID- 16311208 TI - [Autonomic profile of patients with migraine]. AB - OBJECT: Dysfunction of autonomic nervous system (ANS) is implicated in the genesis and persistence of migraine. The objective of this study was to compare autonomic nervous system (ANS) profile of migraineurs during headache-free periods to a group of normal subjects based on cardio-vascular reactivity. METHODS: Patients with migraine according to the criteria of IHS 2004 were selected for the study. After a 30 min resting blood pressure (BP), the following standard tests were performed: deep-breathing (DB), hand grip (HG) of 15 s and 3 min, valsalva maneuver, echo stress, (ES) and tilt test (TT). Results were compared to 44 normal subjects, age similar, 37 female, (84.1%) using the Student test, with P < 0.005 as significant. RESULTS: Thirty-two patients (27 female (84.38%), 16-51 years, mean 40.41 +/- 7.8) were studied. Twenty-two patients (69%) had systolic blood pressure below 94 mmHg and 25 patients (78%) had diastolic blood pressure below 60 mmHg. Compared to normal, migraineurs exhibited a significantly higher vagal response (P < 0.001) and a significantly lower alpha sympathetic response, central by using ES as well as peripheral by using HG of 3 min (P < 0.001). CONCLUSIONS: Autonomic cardiovascular reactivity of patients with migraine showed a vagal hyperactivity and a deficiency of the alpha sympathetic system. This leads to further studies with new therapeutical approaches. PMID- 16311209 TI - A study on visual evoked responses in children with chronic renal failure. AB - AIMS OF THE STUDY: Nervous involvement is frequent in patients with renal failure. Early recognition of the condition by electrophysiological tests may provide means for protective measures before irreversible damage of nervous system (NS) structures takes place. This study has two objectives: (1) examining whether pattern-reversal visual evoked potential (PR-VEP) studies may provide information relating to possible subclinical NS involvement in pediatric patients with chronic renal failure (CRF) and (2) looking for a possible relationship between serum parathormone (PTH) and creatinine levels and PR-VEP parameters. METHODS: PR-VEP recordings at low spatial frequencies were performed and peak-to peak amplitudes and latencies of the P100 component were measured in 19 neurologically asymptomatic children with CRF, 15 of whom were on continuous ambulatory peritoneal dialysis (CAPD) and four on hemodialysis (HD). A similar procedure was applied to 29 healthy, age- and sex-matched, subjects. Patients were sub-grouped according to the serum PTH and creatinine levels. Student's-t and one-way ANOVA tests were used for comparisons within patient and control groups and sub-groups relating to serum PTH and creatinine levels. RESULTS: We did not demonstrate any statistically significant differences in PR-VEP parameters in patients vs. controls. PR-VEP amplitudes were higher in patients with low serum creatinine levels as compared to group with high creatinine values and to controls. No other relationship was found between PR-VEP parameters and serum PTH and creatinine levels in this pediatric population. CONCLUSION: Young patients with CRF and under dialysis do not necessarily show pathologic alterations in PR-VEPs when they are neurologically intact. This fact suggests that either PR-VEPs are not sensitive enough to detect clinically silent NS involvements in such patients, or it could be related to positive effects of the currently improving standards in the management of dialysis and supportive nutrition. Additional factors such as the age of the patient during examination, the latency between dialysis and visual evoked potential (VEP) assessment, or the selected check size may have some impact on the results and justify further studies. PMID- 16311210 TI - Reversible non-metabolic triphasic waves. PMID- 16311212 TI - Human fat cell lipolysis: biochemistry, regulation and clinical role. AB - Release of fatty acids (FAs) from adipose tissue through lipolysis in fat cells is a key event in many processes. FAs are not only energy substrates but also signalling molecules and substrates for lipoprotein production by the liver. Fat cells consist of>95% triglycerides that are hydrolysed during lipolysis to glycerol and FAs. The major rate-limiting factor for lipolysis is hormone sensitive lipase, but additional lipases such as adipose tissue triglyceride lipase may also play a role. The regulation of human fat cell lipolysis is, in many ways, species unique. Only catecholamines, insulin and natriuretic peptides have pronounced acute effects. Catecholamines influence lipolysis through four different adrenoceptor subtypes, in contrast to rodents where only one subtype (beta(3)) is of major importance. There are regional variations in adipocyte lipolysis leading to more release of FAs from the visceral than subcutaneous adipose tissue during hormone stimulation (insulin, catecholamines). Since, only visceral fat is linked to the liver (by the portal vein), alterations in visceral adipocyte tissue lipolysis have direct effects on the liver through portal FA release. The regional variations in lipolysis are further enhanced in obesity and polycystic ovarian syndrome, and are of importance for dyslipidaemia, hyperinsulinaemia and glucose intolerance in these conditions. There is a marked elevation of circulating FA levels among the obese, which may be due to enhanced production of tumour necrosis factor alpha in adipose tissue. This cytokine stimulates lipolysis through so-called MAP kinases. Pharmacological agents in clinical practice such as nicotinic acid and glitazones exert lipid-lowering and glucose-lowering effects, respectively, by decreasing FA output from the adipose tissue. This review covers the biochemistry, regulation and clinical aspects of human fat cell lipolysis. PMID- 16311213 TI - Adipogenesis: cellular and molecular aspects. AB - Obesity and lipoatrophy are major risks for insulin resistance, non-insulin dependent diabetes and cardiovascular disease. In the past three decades, significant advances have been made in delineating the key transcription factors of adipogenesis, as well as extracellular effectors and intracellular signalling pathways that regulate fat cell formation. This review focuses on in vitro models of adipocyte differentiation, and on the balance between pro- and anti-adipogenic factors that drive the adipocyte differentiation process. Full understanding of the mechanisms of adipose tissue differentiation represents a major issue to develop a comprehensive strategy to prevent and treat obesity. PMID- 16311214 TI - Peroxisome proliferator-activated receptor gamma and the regulation of adipocyte function: lessons from human genetic studies. AB - In recent years, the thiazolidinediones (e.g. rosiglitazone, pioglitazone) have emerged as an exciting novel class of therapeutic agent for the treatment of type 2 diabetes mellitus and the human metabolic syndrome. At first glance, the use of these high-affinity peroxisome proliferator-activated receptor gamma (PPARgamma) agonists, that promote adipogenesis, to treat a group of disorders that typically have their origins in obesity seems counter-intuitive. However, to view PPARgamma simply as a regulator of fat mass, and adipocytes themselves as passive vessels for energy storage, is to ignore an extensive body of data that speaks of the diverse roles of both this receptor and adipose tissue in the maintenance of normal metabolic homeostasis. This article highlights the important clinical and laboratory observations made in human subjects harbouring genetic variations in PPARgamma that have confirmed its pivotal role in the regulation of adipocyte endocrine function, and thus our metabolic response to the environment. PMID- 16311215 TI - Adipocytokines: leptin--the classical, resistin--the controversical, adiponectin- the promising, and more to come. AB - With the growing prevalence of obesity, scientific interest in the biology of adipose tissue has been extended to the secretory products of adipocytes, since they are increasingly shown to affect several aspects in the pathogenesis of obesity-related diseases. The cloning of the ob gene is consistent with this concept and suggests that body fat content in adult rodents is regulated by a negative feedback loop centred in the hypothalamus. In recent years, a number of additional signalling molecules secreted by adipose tissue have been discovered, commonly referred to as 'adipocytokines'. Among these, adiponectin is perhaps the most interesting and promising compound for the clinician since it has profound protective actions in the pathogenesis of diabetes and cardiovascular disease. Adiponectin is low in obese subjects and, in particular, insulin-resistant patients. In contrast, resistin seems to be of greater relevance in relation to the immune stress response than in the regulation of glucose homeostasis. However, inflammatory processes have recently been connected with the development of atherosclerosis. Finally, little is known regarding the clinical relevance of visfatin. Recent research has revealed many functions of adipocytokines extending far beyond metabolism, such as immunity, cancer and bone formation. This report aims to review some of the recent topics of adipocytokine research that may be of particular importance. PMID- 16311216 TI - Adipose tissue: a regulator of inflammation. AB - Adipose tissue is a highly active organ. In addition to storing calories as triglycerides, it also secretes a large variety of proteins, including cytokines, chemokines and hormone-like factors, such as leptin, adiponectin and resistin. Intriguingly, many, if not most, of these adipose-derived proteins have dual actions; cytokines have both immunomodulatory functions and act as systemic or auto-/paracrine regulators of metabolism, while proteins such as leptin and adiponectin are regulators of both metabolism and inflammation. The production of pro-atherogenic chemokines by adipose tissue is of particular interest since their local secretion, e.g. by perivascular adipose depots, may provide a novel mechanistic link between obesity and the associated vascular complications. PMID- 16311217 TI - Mineralocorticoid-stimulating activity of adipose tissue. AB - Obesity is strongly associated with arterial hypertension. A positive correlation between obesity and plasma aldosterone levels has been observed by different investigators, suggesting that an abnormal secretion of aldosterone in obesity contributes to the development of arterial hypertension in obese subjects. The mechanisms proposed to explain this abnormal aldosterone production mainly involve the adipose renin-angiotensin system, an indirect effect of increased fatty acids, and direct adrenal stimulation by adipocyte secretory products. Indeed, adipose mineralocorticoid-stimulating activity was recently observed in isolated human adipocytes, suggesting a hitherto unknown direct involvement of adipose tissue in the regulation of blood pressure in obesity. PMID- 16311218 TI - Insulin-like growth factor I, growth hormone and insulin in white adipose tissue. AB - Maturation of adipose tissue results from both the expansion of mature adipocytes and the formation of new adipocytes from adipocyte precursor cells. A variety of hormones related to adipogenesis have been identified recently. Both growth hormone (GH) and insulin-like growth factor I (IGF-I) are of major significance in adipocyte differentiation. IGF-I has been suggested to be a major regulator of cell proliferation, differentiation and metabolism, thus regulating, among other biological processes, adipose tissue growth and differentiation of pre-adipocytes into adipocytes. GH exerts its effects by increasing the pool of adipocyte precursor cells capable of differentiating into mature adipocytes. In addition, GH seems to have the potential to reduce the volume of mature adipocytes, thus inhibiting the expansion of adipose tissue and reducing body fat. This chapter gives an overview of studies that have investigated the roles of insulin, GH and IGF-I in adipogenesis. PMID- 16311219 TI - Nutrition-/diet-induced changes in gene expression in white adipose tissue. AB - Nutrients regulate metabolic fluxes and homeostasis through transcriptional and translational control of enzyme concentrations and allosteric modulation of enzyme activity. Dietary omega-3 polyunsaturated fatty acids (PUFAs) have been shown to exert a variety of beneficial health effects such as reducing adiposity and increasing insulin sensitivity in rodents. It is now clear that PUFAs regulate fundamental adipose cell and liver functions through modulation of activity and abundance of key transcription factors that act as nutrient sensors, including peroxisome proliferator-activated receptors (PPARalpha/delta/gamma), sterol regulatory element binding proteins (SREBP-1/2), and liver X receptors (LXRalpha/beta). However, in the state of obesity, where adipose tissue shows elevated storage of triglycerides, many lipogenic genes that are essential for adipose cell function including PPARgamma, SREBP-1c, CCAAT-enhancer binding protein alpha and stearoyl-CoA desaturase-1 are downregulated, apparently due to desensitization of the very same crucial nutrient sensors. This chapter will summarize recent studies of PUFA- and obesity-induced changes in gene expression in white adipose tissue. PMID- 16311220 TI - Transgenic animal models for the study of adipose tissue biology. AB - The traditional view of adipose tissue as a passive energy reservoir has changed. Adipose tissue is a complex, highly active metabolic and endocrine organ. With obesity as an increasingly important public health threat, a major development in the understanding of adipose tissue biology has come with observations in different biological spheres including whole-body physiology and application of transgenic animal models. Scientific progress has been made with the identification of several genes in spontaneous monogenic animal models of obesity, and in understanding the molecular mechanisms underlying phenotypes of altered body weight, adiposity and fat distribution by creating transgenic and knockout animal models. Mouse phenotypes resulting from inactivation or overexpression of molecules responsible for the regulation of adipose tissue metabolism have led to novel concepts in the understanding of adipocyte biology and development of obesity. This review presents an overview of transgenic animal models for the study of adipose tissue biology. PMID- 16311221 TI - Fatty acids and insulin resistance in muscle and liver. AB - Free fatty acids (FFAs) circulate round the body and represent important nutrients and the key oxidative fuel for the heart and resting skeletal muscle. In addition, FFAs are thought to be potent signalling molecules. Growing evidence indicates that FFAs may be involved in type 2 diabetes mellitus and obesity by mediating insulin resistance. In 1963, it was postulated that accumulated glucose 6-phosphate as a result of increased FFA oxidation leads to decreased glucose uptake. An alternative hypothesis is that increased concentrations of plasma FFA induce insulin resistance in humans through inhibition of glucose transport activity, which appears to be a consequence of decreased insulin receptor substrate-1-associated phosphatidyl inositol 3 kinase activity. Moreover, FFAs can arise locally, and increased intramyocellular and hepatocellular lipids have been shown to be associated with insulin resistance. This paper reviews the main aspects of FFA metabolism in the development of insulin resistance in skeletal muscle and liver, as well as the role of ectopic lipid deposits as a local source of FFAs. Finally, the role of thiazolidinediones as modulators of FFA-induced insulin resistance will be discussed. PMID- 16311222 TI - White adipose tissue and cardiovascular disease. AB - Adipocytes have recently been shown to secrete a variety of bioactive substances called 'adipocytokines', and have been recognized as endocrine cells. Tumour necrosis factor (TNF)-alphaalpha, plasminogen activator inhibitor-1 (PAI-1) and heparin-binding epidermal-growth-factor-like growth factor (HBEGF) are among these adipocytokines, and they contribute to the development of vascular diseases. Visfatin is a visceral fat-specific protein that may be related to the development of obesity-related diseases such as diabetes mellitus and cardiovascular disease. In contrast, adiponectin, an adipose-tissue-specific collagen-like protein, has recently been reported as an important anti atherogenic and anti-diabetic protein. Adipocytokine secretion may be regulated dynamically by the nutritional state. Visceral fat accumulation leads to dysfunction of adipocytes (including hypersecretion of TNF-alphaalpha, PAI-1 and HBEGF, and hyposecretion of adiponectin), which results in the development of a variety of metabolic and circulatory diseases. In this review, the importance of adipocytokines, including adiponectin, is discussed with respect to cardiovascular disease. PMID- 16311223 TI - Link between obesity and type 2 diabetes. AB - The relationship between obesity and diabetes is of such interdependence that the term 'diabesity' has been coined. The passage from obesity to diabetes is made by a progressive defect in insulin secretion coupled with a progressive rise in insulin resistance. Both insulin resistance and defective insulin secretion appear very prematurely in obese patients, and both worsen similarly towards diabetes. Thus, the classic 'hyperbolic relationship' between insulin resistance and insulin secretion and the 'glucose allostasis concept' remain prevailing concepts in this particular field of knowledge. An increase in overall fatness, preferentially of visceral as well as ectopic fat depots, is specifically associated with insulin resistance. The accumulation of intramyocellular lipids may be due to reduced lipid oxidation capacity. The ability to lose weight is related to the capacity to oxidize fat. Thus, a relative defect in fat oxidation capacity is responsible for energy economy and hampered weight loss. PMID- 16311226 TI - Reconciling cultural relativism for a clinical paradigm: what's a nurse to do? PMID- 16311227 TI - What's in a name? Multidisciplinary, interdisciplinary, and transdisciplinary. PMID- 16311228 TI - Transforming health care from the inside out: advancing evidence-based practice in the 21st century. AB - Health care is in need of change. Major professional and health care organizations as well as federal agencies and policy-making bodies are emphasizing the importance of evidence-based practice (EBP). Using this problem solving approach to clinical care that incorporates the conscientious use of current best evidence from well designed studies, a clinician's expertise, and patient values and preferences, nurses and other health care providers can provide care that goes beyond the status quo. Health care that is evidence-based and conducted in a caring context leads to better clinical decisions and patient outcomes. Gaining knowledge and skills in the EBP process provides nurses and other clinicians the tools needed to take ownership of their practices and transform health care. Key elements of a best practice culture are EBP mentors, partnerships between academic and clinical settings, EBP champions, clearly written research, time and resources, and administrative support. This article provides an overview of EBP and offers recommendations for accelerating the adoption of EBP as a culture in education, practice and research. PMID- 16311229 TI - Educating for evidence-based practice. AB - Several countries have developed policy initiatives that require health care to be evidence based and health science students to study under an evidence-based curriculum as well as learn how to practice in an evidence-based way. Evidence based nursing (EBN) is too new to have any actual research on which to base a curriculum. However, in the absence of evidence, several schools of nursing have moved or are moving forward. This article presents ideas and examples gleaned from colleagues around the world on the why's and how's of teaching EBN. Finally, this article calls for the funding and planning of an evaluation of evidence based curricula and teaching strategies for continued development of the field. PMID- 16311230 TI - Building a foundation for an evidence-based approach to practice: teaching basic concepts to undergraduate freshman students. AB - The University of Pittsburgh School of Nursing introduced evidence-based practice (EBP) into the freshman-level curriculum this year. This article describes the reasons behind this decision, the development of the curriculum, the specific EBP content, and the student assignments. The introduction of EBP into the first level nursing curriculum has proven to be successful; consideration is even being given to expanding the EBP content. PMID- 16311231 TI - Developing research competence to support evidence-based practice. AB - This article describes one step in the process that was undertaken to prepare for the introduction of evidence-based practice (EBP) into the curriculum across the Bachelor of Science in Nursing, Master of Science in Nursing, and Doctor of Philosophy programs, as well as the programs that were under development, Clinical Nurse Leader and Doctor of Nursing Practice, at the University of Pittsburgh School of Nursing. Expected research competencies were identified for each level or academic year within each program. Based on these competencies, recommendations on how to modify the curriculum into one that would support students' acquisition and development of the skills necessary to be successful in matriculating through an EBP curriculum were developed. Evaluation mechanisms for the achievement of these competencies vary across the academic programs and will include performance on capstone projects, comprehensive examinations, and program milestones for doctoral students. The establishment of evidence-based competencies provided a foundation for the development of new teaching approaches and the curricular revisions across the three academic programs. Thus, the University of Pittsburgh model of educating for EBP is based on a sequential layering of research competencies throughout the curriculum. PMID- 16311232 TI - Critical thinking and evidence-based practice. AB - Critical thinking (CT) is vital to evidence-based nursing practice. Evidence based practice (EBP) supports nursing care and can contribute positively to patient outcomes across a variety of settings and geographic locations. The nature of EBP, its relevance to nursing, and the skills needed to support it should be required components of baccalaureate education and must be introduced early in students' development as independent, self-directed learners and as professional nurses. Among the knowledge, skills, and processes needed to support EBP, CT is paramount. The development of CT can prepare nurses with the necessary skills and dispositions (habits of mind, attitudes, and traits) to support EBP. The intents of this study were to explore the importance of CT as an essential skill to support EBP and to describe some of the strategies and processes considered key to the ongoing development of CT. PMID- 16311233 TI - Toward clinical scholarship: promoting evidence-based practice in the clinical setting. AB - Organizational interventions are being suggested to increase the rate of quality research dissemination and uptake. This article describes how one tertiary institution is using an evidence-based nursing (EBN) committee as an organizational strategy to shift its nursing culture toward clinical scholarship. A number of approaches and activities that have stimulated the movement toward evidence-based practice (EBP) are examined: organizational commitment to EBP, strategic positioning of the EBN committee within nursing's administrative structure, articulation of a mission, conceptualization of a model for EBN practice, learning on the job, selection and adoption of an evidence-based model for implementing change, marketing for a change in culture toward clinical scholarship, and other selected examples of projects undertaken by the committee. Action-oriented principles associated with committee experiences are related to the approaches and activities. PMID- 16311234 TI - Evidence-based resources and the role of librarians in developing evidence-based practice curricula. AB - The implementation of evidence-based practice (EBP) requires acquisition and use of a complex set of skills, including the ability to locate and critically evaluate clinically relevant research literature. In this article, we discuss information resources and tools that may be of value to educators faced with the task of teaching students to search for and evaluate research-based evidence. In addition, we discuss how health sciences librarians, with the use of new models of information instruction and delivery, can work with nursing faculty in developing curricula for training students in EBP. PMID- 16311235 TI - How to overcome difficulties in the evaluation of best possible decision-making in redo-revascularization? PMID- 16311236 TI - Role of angiotensin system and effects of its inhibition in atrial fibrillation: clinical and experimental evidence. AB - Atrial fibrillation (AF) is a common arrhythmia that is difficult to treat. Anti arrhythmic drug therapy, to maintain sinus-rhythm, is limited by inadequate efficacy and potentially serious adverse effects. There is increasing interest in novel therapeutic approaches that target AF-substrate development. Recent trials suggest that angiotensin converting-enzyme (ACE)-inhibitors and angiotensin receptor blockers (ARBs) may be useful, particularly in patients with left ventricular hypertrophy or failure. The clinical potential and mechanisms of this approach are under active investigation. Angiotensin-II is involved in remodelling and may have direct electrophysiological actions. Experimental studies show protection from atrial structural and possibly electrical remodelling with ACE-inhibitors and ARBs, as well as potential effects on cardiac ion-channels. This article reviews information pertaining to the clinical use and mechanism of action of ACE-inhibitors and ARBs in AF. A lack of prospective randomized double-blind trials data limits their application in AF patients without another indication for their use, but studies under way may alter this in the near future. This exciting field of investigation may lead to significant improvements in therapeutic options for AF patients. PMID- 16311237 TI - Implementation of reperfusion therapy in acute myocardial infarction. A policy statement from the European Society of Cardiology. AB - Reperfusion therapy in ST-segment elevation myocardial infarction (STEMI) is the most important component of treatment, as it strongly influences short- and long term patient outcome. The main objective of healthcare providers should be to achieve at least 75% of reperfusion therapy applied to patients suffering from STEMI in a timely manner, and preferably within the first 3 h after onset of symptoms. Establishing networks of reperfusion at regional and national level, implying close collaboration between all the actors involved in reperfusion therapy, namely hospitals, departments of cardiology, PCI centres, emergency medical systems (EMS), (para)medically staffed ambulances, private cardiologists, primary care physicians, etc., is a key issue. All forms of reperfusion, depending on local facilities, need to be available to patients. Protocols must be written and agreed for the strategy of reperfusion to be applied within a network. Early diagnosis of STEMI is essential and is best achieved by rapid ECG recording and interpretation at first medical contact, wherever this contact takes place (hospital or ambulance). Tele-transmission of ECG for immediate interpretation by experienced cardiologists is an alternative. Primary PCI is the preferred reperfusion option if it can be performed by experienced staff within 90 min after first medical contact. Thrombolytic treatment, administered if possible in the pre-hospital setting, is a valid option if PCI cannot be performed in a timely manner, particularly within the first 3 h following onset of symptoms. Thrombolysis is not the end of the reperfusion therapy. Rescue PCI must be performed in the case of thrombolysis failure. Next-day PCI after successful thrombolysis has been proven efficacious. Quality control is important for monitoring the efficacy of networks of reperfusion. All elements that influence time to reperfusion must be taken into account, particularly transfer delays, in-hospital delays, and door-to-balloon or door-to-needle times. The rate of reperfusion achieved must also be taken into consideration. Professional organizations such as the European Society of Cardiology (ESC) have the responsibility to impart this message to the cardiology community, and inform politicians and health authorities about the best possible strategy to achieve reperfusion therapy. PMID- 16311238 TI - Heart failure, atrial fibrillation, and beta-blockers. PMID- 16311239 TI - Percutaneous PFO closure, further data but many unanswered questions. PMID- 16311240 TI - Differential sensitivity of the cystic fibrosis (CF)-associated mutants G551D and G1349D to potentiators of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel. AB - The genetic disease cystic fibrosis (CF) is caused by loss of function of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel. Two CF mutants, G551D and G1349D, affect equivalent residues in the highly conserved LSGGQ motifs that are essential components of the ATP-binding sites of CFTR. Both mutants severely disrupt CFTR channel gating by decreasing mean burst duration (MBD) and prolonging greatly the interburst interval (IBI). To identify small molecules that rescue the gating defects of G551D- and G1349D-CFTR and understand better how these agents work, we used the patch clamp technique to study the effects on G551D- and G1349D-CFTR of phloxine B, pyrophosphate (PP(i)), and 2' deoxy ATP (2'-dATP), three agents that strongly enhance CFTR channel gating. Phloxine B (5 microm) potentiated robustly G551D-CFTR Cl- channels by altering both MBD and IBI. In contrast, phloxine B (5 microm) decreased the IBI of G1349D CFTR, but this effect was insufficient to rescue G1349D-CFTR channel gating. PP(i) (5 mm) potentiated weakly G551D-CFTR and was without effect on the G1349D CFTR Cl- channel. However, by altering both MBD and IBI, albeit with different efficacies, 2'-dATP (1 mm) potentiated both G551D- and G1349D-CFTR Cl- channels. Using the ATP-driven nucleotide-binding domain dimerization model of CFTR channel gating, we suggest that phloxine B, PP(i) and 2'-dATP alter channel gating by distinct mechanisms. We conclude that G551D- and G1349D-CFTR have distinct pharmacological profiles and speculate that drug therapy for CF is likely to be mutation-specific. PMID- 16311241 TI - Reactive lipid species from cyclooxygenase-2 inactivate tumor suppressor LKB1/STK11: cyclopentenone prostaglandins and 4-hydroxy-2-nonenal covalently modify and inhibit the AMP-kinase kinase that modulates cellular energy homeostasis and protein translation. AB - LKB1, a unique serine/threonine kinase tumor suppressor, modulates anabolic and catabolic homeostasis, cell proliferation, and organ polarity. Chemically reactive lipids, e.g. cyclopentenone prostaglandins, formed a covalent adduct with LKB1 in MCF-7 and RKO cells. Site-directed mutagenesis implicated Cys210 in the LKB1 activation loop as the residue modified. Notably, ERK, JNK, and AKT serine/threonine kinases with leucine or methionine, instead of cysteine, in their activation loop did not form a covalent lipid adduct. 4-Hydroxy-2-nonenal, 4-oxo-2-nonenal, and cyclopentenone prostaglandin A and J, which all contain alpha,beta-unsaturated carbonyls, inhibited the AMP-kinase kinase activity of cellular LKB1. In turn, this attenuated signals throughout the LKB1 --> AMP kinase pathway and disrupted its restraint of ribosomal S6 kinases. The electrophilic beta-carbon in these lipids appears to be critical for inhibition because unreactive lipids, e.g. PGB1, PGE2, PGF2alpha, and TxB2, did not inhibit LKB1 activity (p > 0.05). Ectopic expression of cyclooxygenase-2 and endogenous biosynthesis of eicosanoids also inhibited LKB1 activity in MCF-7 cells. Our results suggested a molecular mechanism whereby chronic inflammation or oxidative stress may confer risk for hypertrophic or neoplastic diseases. Moreover, chemical inactivation of LKB1 may interfere with its physiological antagonism of signals from growth factors, insulin, and oncogenes. PMID- 16311242 TI - Using qualitative methodology to elucidate themes for a traditional tooth gauging education tool for use in a remote Ugandan community. AB - The gauging of unerupted deciduous canine teeth occurs in approximately one in three children in some areas of Uganda. It is believed that such teeth are 'maggots' that cause fever, diarrohea and vomiting. Traditional healers use knitting needles, bicycle spokes, scissors or broken glass to extract the teeth. Post-ebino extraction complications include septicaemia, anaemia, difficulties in feeding and pain. Some children require hospitalization. Health is further compromised when multiple ebino extractions occur at one time, increasing the risk of human immunodeficiency virus/acquired immunodeficiency syndrome transmission. An ebino education initiative was developed in the southwest Ugandan province of Rukungiri, based on the findings of five community-based focus group discussions. The initiative involved role-plays, didactic presentations and discussion/debate workshops to 23 women's groups in 15 communities (total number of women exposed = 1874). After 1.5 years of the programme's inception, community awareness of the scientific causes and alternatives to ebino extractions had increased (as gauged by follow-up focus group discussion findings) and the number of hospital admissions for traditional tooth extraction complications had reduced. The appropriateness of the model in exploring and addressing ebino extraction beliefs and attitudes is discussed, as are implications of the strategy in its implementation in other communities where ebino extractions are prevalent. PMID- 16311243 TI - Concordance of multiple analytical approaches demonstrates a complex relationship between DNA repair gene SNPs, smoking and bladder cancer susceptibility. AB - Study results of single nucleotide polymorphisms (SNPs) and cancer susceptibility are often conflicting, possibly because of the analytic challenges of testing for multiple genetic and environmental risk factors using traditional analytic tools. We investigated the relationship between DNA repair gene SNPs, smoking, and bladder cancer susceptibility in 355 cases and 559 controls enrolled in a population-based study of bladder cancer in the US. Our multifaceted analytical approach included logistic regression, multifactor dimensionality reduction, and hierarchical interaction graphs for the analysis of gene-gene and gene environment interactions followed by linkage disequilibrium and haplotype analysis. Overall, we did not find an association between any single DNA repair gene SNP and bladder cancer risk. We did find a marginally significant elevated risk of the XPD codon 751 homozygote variant among never smokers [adjusted odds ratio (OR) 2.5, 95% confidence interval (CI) 1.0-6.2]. In addition, the XRCC1 194 variant allele was associated with a reduced bladder cancer risk among heavy smokers [adjusted OR 0.4, 95% CI 0.2-0.9)]. The best predictors of bladder cancer included the XPD codon 751 and 312 SNPs along with smoking. Interpretation of this multifactor model revealed that the relationship between the XPD SNPs and bladder cancer is mostly non-additive while the effect of smoking is mostly additive. Since the two XPD SNPs are in significant linkage disequilibrium (D' = 0.52, P = 0.0001), we estimated XPD haplotypes. Individuals with variant XPD haplotypes were more susceptible to bladder cancer [e.g. adjusted OR 2.5, 95% CI 1.7-3.6] and the effect was magnified when smoking was considered. These results support the hypothesis that common polymorphisms in DNA repair genes modify bladder cancer risk and emphasize the need for a multifaceted statistical approach to identify gene-gene and gene-environment interactions. PMID- 16311244 TI - Genotypes and haplotypes of matrix metalloproteinase 1, 3 and 12 genes and the risk of lung cancer. AB - The MMPs (matrix metalloproteinases) are a family of secreted zinc metalloproteases that degrade the collagens of the extracellular matrix important in tissue remodeling and repair during development and inflammation. We investigated the associations between polymorphisms of MMP-1 (-1607 1G/2G, rs1799750), MMP-3 (-1171 5A/6A, rs3025058), and MMP-12 (-82AG, rs2276109, and 1082A/G, rs652438) and the risk of lung cancer in 2014 Caucasian lung cancer patients and 1323 healthy controls. The results were analyzed using logistic regression models, adjusting for covariates. The four polymorphisms were in Hardy Weinberg disequilibrium. Except for the 1G-1082A, the other linkage disequilibrium tests between the four MMP polymorphisms were statistically significant (P < 0.001). There was no overall association between individual MMP polymorphism and the risk of lung cancer. The MMP polymorphisms jointly were associated with a non-statistically significant higher risk of lung cancer, with the adjusted odds ratio (AOR) of subjects with 5+ variant alleles versus zero variant allele of 1.31 [95% confidence interval (CI), 0.92-1.88]. Stronger associations were observed in never-smokers and males, with the corresponding AORs of 2.44 (95%CI, 1.10-5.43, P(trend) = 0.04) in never smokers and 1.35 (95%CI, 0.79-2.30, P(trend) = 0.04) in men. In haplotype analysis, the 1G-6A-82A 1082G haplotype was associated with higher risk of lung cancer among never smokers, with the AOR of 3.65 (95%CI, 1.62-8.20) when compared with the most common 1G-5A-82A-1082A haplotype. In conclusion, the combined MMP genotypes and associated haplotypes may be associated with higher risk of lung cancer, particularly among never smokers and men. PMID- 16311245 TI - Overexpression of cyclins D1 and D3 during estrogen-induced breast oncogenesis in female ACI rats. AB - A common feature of human breast oncogenesis is cell cycle deregulation. The expression of cyclins D1 and D3 was examined during estradiol-17beta (E(2)) induced mammary tumorigenesis in female August Copenhagen Irish (ACI) rats. Low serum E(2) levels ( approximately 60-120 pg/ml) were sufficient to induce mammary gland tumors (MGTs) that remarkably resemble human ductal breast cancer (BC) at the histopathologic and molecular levels. Western blot analysis of the E(2) induced MGTs revealed a marked rise in cyclins D1 (24-fold), D3 (9-fold) and cdk4 (3-fold) expression compared with age-matched untreated controls. Small focal dysplasias with large, pale staining nuclei were commonly seen at 3-3.6 months, large focal dysplasias, including atypical ductal hyperplasia at 3.6-4.3 months, ductal carcinoma in-situ (DCISs) at 4.3-5.0 months, and 100% incidence of invasive ductal BC/frank tumors at 5-6 months were detected after E(2) treatment. Immunohistochemical analysis of serial sections of focal dysplasias, DCISs and invasive ductal carcinomas showed overexpression of cyclins D1, D3, estrogen receptor-alpha (ERalpha) and progesterone receptor (PR). However, cyclin D3 expression, unlike D1, was confined essentially to early pre-malignant lesions (focal dysplasias and DCISs) and primary MGTs with <1-5% of resting and normal hyperplastic breast cells staining positive. The kinase activity for cyclins D1 and D3, using retinoblastoma (Rb) as a substrate, in E(2)-induced MGTs and their binding to cdk4 was significantly elevated. Semi-quantitative reverse transcriptase PCR analysis of the E(2)-induced MGTs exhibited increased expression of cyclins D1 (2.9-fold) and D3 (1.4-fold) mRNA, indicating that their elevated protein expression was due in part to an increase in mRNA transcription. However, when analyzed by quantitative real-time Q-PCR, these genes were not amplified. These data indicate that in female ACI rat mammary glands, E(2) induced pre-malignant lesions differentially and selectively express cyclins D1 and D3, thus contributing to a distinct growth advantage of these pre-neoplasias relative to E(2)-elicited normal hyperplasia. PMID- 16311246 TI - Green tea, black tea and breast cancer risk: a meta-analysis of epidemiological studies. AB - Experimental studies have shown that tea and tea polyphenols have anti carcinogenic properties against breast cancer. A number of epidemiologic studies, both case-control and cohort in design, have examined the possible association between tea intake and breast cancer development in humans. This meta-analysis included 13 papers which examined populations in eight countries and provided data on consumption of either green tea or black tea, or both in relation to breast cancer risk. Summary odds ratios (ORs) for highest versus non/lowest tea consumption level were calculated based on fixed and random effects models. Heterogeneity between studies was examined via the Q statistics. For green tea, the combined results from the four studies indicated a reduced risk of breast cancer for highest versus non/lowest intake (OR = 0.78, 95% CI = 0.61-0.98). For black tea, conflicting results were observed in case-control versus cohort studies. The combined results from the eight case-control studies showed a minor inverse association between black tea consumption and risk of breast cancer (OR = 0.91, 95% CI = 0.84-0.98). This inverse association was stronger in hospital based (OR = 0.77, 95% CI = 0.50-1.19) than population-based case-control studies (OR = 0.94, 95% CI = 0.81-1.09). Five cohort studies demonstrated a modest increase in risk associated with black tea intake (OR = 1.15, 95% CI = 1.02 1.31). The results of this meta-analysis indicate a lower risk for breast cancer with green tea consumption. Available data suggest a possible late-stage, promotional effect of black tea on breast carcinogenesis. PMID- 16311247 TI - It's in your hands: the value of handsearching in conducting systematic reviews of public health interventions. AB - BACKGROUND: While there is an emerging evidence base in public health, the evidence can often be difficult to find. Indexing of journals in MEDLINE has assisted those conducting systematic reviews to more easily identify published studies. However, information technology and the processes associated with indexing are not infallible. Studies may not be correctly marked by study design which may mean they are missed in the electronic searching process. Handsearching for evidence of intervention effectiveness has therefore become a recognized tool in the systematic review process. METHODS: Resources to guide handsearching activity currently are clinically focused, and may not be sensitive to the characteristics of public health studies where study terminology may differ. In response to this issue, the Cochrane Health Promotion and Public Health Field (the Field) developed and implemented a small study to recruit and support handsearchers from around the world to identify health promotion and public health trials and systematic reviews. A strategic framework was developed to recruit and support handsearchers to search six public health-related journals. RESULTS: In total, 131 trials and 21 systematic reviews were identified. The greatest value of handsearching was found to be in supplement editions and abstract sections of journals CONCLUSIONS: The study focused exclusively on indexed journals with the intention that tools and methods developed could be used to explore the potential for handsearching in non-indexed journals and for unpublished studies. The findings from this study will continue to support handsearching efforts and in doing so contribute to high quality systematic reviews of public health interventions. PMID- 16311248 TI - Communicable disease and health protection quarterly review: April to June 2005: from the health protection agency, centre for infections. PMID- 16311249 TI - Dental filling as an indicator of socio-economic status. PMID- 16311250 TI - Dr. Laidlaw's prescription for the homeless. PMID- 16311251 TI - Intracerebral adeno-associated virus-mediated gene transfer in rapidly progressive forms of metachromatic leukodystrophy. AB - Metachromatic leukodystrophy (MLD) is a neurodegenerative lysosomal disease caused by a defect of the enzyme arylsulfatase A (ARSA) that disrupts the degradation of sulfatides (Sulf) in neurons and glial cells. Therapy for MLD requires active production of ARSA in the brain to prevent demyelination and neuronal damage, and efficient delivery of ARSA to act faster than disease progression, particularly in the rapidly progressive late infantile form. We used an adeno-associated virus serotype 5 (AAV5) vector to express the human ARSA gene in the brain of MLD mouse model. We achieved rapid, extensive and long-lasting expression of the recombinant ARSA in the brain, cerebellum and brainstem from at least 3 to 15 months post-injection. Analysis of the vector genome and ARSA distribution gave evidence for in vivo cross-correction of many untransduced neurons and astrocytes. ARSA delivery rapidly reversed Sulf storage and prevented neuropathological abnormalities and neuromotor impairment. We believe that AAV5 mediated brain delivery of ARSA is a potentially efficacious therapeutic strategy for MLD patients, especially for those with rapidly progressive form of the disease. PMID- 16311252 TI - Telomere instability in the male germline. AB - Telomeres play a key role in upholding the integrity of the genome, and telomerase expression in spermatogonial stem cells is responsible for the maintenance of telomere length in the human male germline. We have previously described extensive allelic variation in somatic cell telomere length that is set in the zygote, the ultimate source of which may be the germline. This implies that despite telomerase activity, substantial telomere length variation can be generated and tolerated in the germline; in order to investigate this further, we have examined the nature of telomere length variation in the human male germline. Here, we describe an analysis of both genome-wide telomere length and single molecule analysis of specific chromosome ends in human sperm. We observed individual specific differences in genome-wide telomere length. This variation may result from genetic differences within the components that determine the telomere length setting of each individual. Superimposed on the genome wide telomere length setting was a stochastic component of variation that generates germ-cells containing severely truncated telomeres. If not re-lengthened during early embryogenesis, such telomeres may limit the replicative capacity of cells derived from the zygote and have the potential to create fusagenic chromosomes, unbalanced translocations and terminal micro-deletions. These data may have implications for the genetic determination of ageing, genetic disease and fertility. PMID- 16311253 TI - Proteasome impairment does not contribute to pathogenesis in R6/2 Huntington's disease mice: exclusion of proteasome activator REGgamma as a therapeutic target. AB - Huntington's disease (HD) is one of a group of neurodegenerative disorders caused by the pathological expansion of a glutamine tract. A hallmark of these so-called polyglutamine diseases is the presence of ubiquitylated inclusion bodies, which sequester various components of the 19S and 20S proteasomes. In addition, the ubiquitin-proteasome system (UPS) has been shown to be severely impaired in vitro in cells overexpressing mutant huntingtin. Thus, because of its fundamental housekeeping function, impairment of the UPS in neurons could contribute to neurotoxicity. We have recently proposed that the proteasome activator REGgamma could contribute to UPS impairment in polyglutamine diseases by suppressing the proteasomal catalytic sites responsible for cleaving Gln-Gln bonds. Capping of proteasomes with REGgamma could therefore contribute to a potential 'clogging' of the proteasome by pathogenic polyglutamines. We show here that genetic reduction of REGgamma has no effect on the well-defined neurological phenotype of R6/2 HD mice and does not affect inclusion body formation in the R6/2 brain. Surprisingly, we observe increased proteasomal 'chymotrypsin-like' activity in 13 week-old R6/2 brains relative to non-R6/2, irrespective of REGgamma levels. However, assays of 26S proteasome activity in mouse brain extracts reveal no difference in proteolytic activity regardless of R6/2 or REGgamma genotype. These findings suggest that REGgamma is not a viable therapeutic target in polyglutamine disease and that overall proteasome function is not impaired by trapped mutant polyglutamine in R6/2 mice. PMID- 16311254 TI - Trehalose reduces aggregate formation and delays pathology in a transgenic mouse model of oculopharyngeal muscular dystrophy. AB - Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant disease that presents in the fifth or sixth decade with dysphagia, ptosis and proximal limb weakness. OPMD is caused by the abnormal expansion of a polyalanine tract within the coding region of polyA binding protein nuclear 1 (PABPN1). The resultant mutant PABPN1 forms aggregates within the nuclei of skeletal muscle fibres. We have previously described a transgenic mouse model of OPMD that recapitulates the human disease and develops progressive muscle weakness accompanied by the formation of aggregates in skeletal muscle nuclei. The chemical chaperone trehalose has been used effectively to alleviate symptoms in a mouse model of Huntington's disease and is thought to elicit its effect by binding and stabilizing partially folded polyglutamine proteins and inhibiting the formation of aggregates. Here, we show that trehalose reduces aggregate formation and toxicity of mutant PABPN1 in cell models. Furthermore, oral administration of trehalose attenuated muscle weakness, reduced aggregate formation and decreased the number of TUNEL-labelled nuclei in skeletal muscle in an OPMD transgenic mouse model. Thus, anti-aggregation therapy may prove effective in the treatment of human OPMD. PMID- 16311255 TI - Mutagenesis studies with four stereoisomeric N2-dG benzo[a]pyrene adducts in the identical 5'-CGC sequence used in NMR studies: G->T mutations dominate in each case. AB - Benzo[a]pyrene (B[a]P) is a polycyclic aromatic hydrocarbon (PAH) and a potent mutagen/carcinogen found ubiquitously in the environment. B[a]P is primarily metabolized to diol epoxides, which react principally at N2-dG in DNA. B[a]P-N2 dG adducts have been shown to induce a variety of mutations, notably G-->T, G- >A, G-->C and -1 frameshifts. Four stereoisomers of B[a]P-N2-dG (designated: [+ta]-;, [+ca]-, [-ta] and [-ca]) were studied by NMR in duplex 11mers in a 5' CGC sequence context, and each adopted a different adduct conformation (Geacintov, et al. (1997) Chem. Res. Toxicol., 10, 111). Herein these four identical B[a]P-containing 11mers are built into duplex plasmid genomes and mutagenesis studied in Escherichia coli following SOS-induction. In nucleotide excision repair (NER) proficient E.coli, no adduct-derived mutants are detected. In NER deficient E.coli, G-->T mutations dominate for all four stereoisomers [+ta]-, [+ca]-, [-ta] and [-ca]-B[a]P-N(2)-dG, and mutation frequency is similar. Thus, the mutagenic pattern for these four B[a]P-N2-dG stereoisomers is the same, in spite of the fact that they adopt dramatically different conformations in ds oligonucleotides as determined by NMR. These findings suggest that adduct conformation must be fluid enough in the 5'-CGC sequence that the duplex DNA conformation can interconvert to mutagenic and non-mutagenic conformations during lesion-bypass. A comparison of all published studies with these four B[a]P-N2-dG stereoisomers in E.coli reveals that B[a]P-N2-dG adduct stereochemistry tends to have a lesser impact on mutagenic pattern (e.g. G-->T versus G-->A mutations) than does DNA sequence context, which is discussed. PMID- 16311256 TI - A 14-year-old girl with renal abnormalities after brief intrauterine exposure to enalapril during late gestation. PMID- 16311257 TI - Treatment of severe steroid-dependent nephrotic syndrome (SDNS) in children with tacrolimus. AB - BACKGROUND: Severe steroid-dependent nephrotic syndrome (SDNS) is a common type of nephrotic syndrome (NS) observed in childhood. Steroid-sparing agents such as calcineurin inhibitors (CNIs) are used to avoid steroid toxicity in SDNS. Tacrolimus (TAC) has been prescribed for maintaining remission of NS in patients who have developed treatment resistance or adverse effects with cyclosporin A (CYA) at our institution since 1995. The aim of this study was to compare the efficacy and complications of TAC with CYA in the management of severe SDNS. METHODS: We report a retrospective longitudinal clinical series of patients with SDNS, all of whom have been treated with TAC. RESULTS: Ten SDNS children (eight males) were reviewed quarterly from time of initial referral to the present day during 93 completed treatment patient years. Nine patients had minimal change disease and one had focal segmental glomerulosclerosis on their first biopsy. The median age at diagnosis was 2.9 years (range 1.6-12.9). The median age at initial referral was 3.9 years (range 2.2-12.9). All patients initially responded to prednisolone at 60 mg/m2/day, and subsequent frequent relapses were treated sequentially with oral cyclophosphamide 168 mg/kg over 8-12 weeks (n = 10), CYA (n = 10), intravenous mustine or a second course of cyclophosphamide (n = 7) and then TAC (n = 10). The initial daily treatment of CYA and TAC in two divided doses was 5 and 0.1 mg/kg/day, respectively; targeted 12 h blood drug levels were of 50-100 microg/l for CYA and 5-10 microg/l for TAC. Patients underwent renal biopsy and the formal glomerular filtration rate (GFR) was measured using plasma clearance of the Inutest method every 2-3 years while receiving CNIs. Six patients continued with TAC; in four patients, TAC was discontinued because of poor response (n = 2), hypertension (n = 1) and glucose intolerance (n = 1). For CYA and TAC treatment periods, the median NS relapse rate was two and one relapses per year, respectively, and cumulative steroid dosage was 73.9 and 105.2 mg/kg/day, respectively (P = 0.54). The reduction in GFR was 5.8 and 11.7 ml/min/1.73 m2 during these periods. Three of the 10 patients showed histological evidence of mild CNI nephrotoxicity over the whole of the CNI treatment period despite achieving target therapeutic drug levels; no significant change in measured or calculated GFR over this prolonged CNI therapy was observed. Antihypertensive medication was prescribed for 11 of 31 CYA and 22 of 40 TAC treatment years. Growth was maintained during the entire CNI therapy period with median change in height SD scores (SDS) of +0.37 and -0.03 over CYA and TAC, respectively (P = 0.13). CONCLUSION: In conclusion, we observed that the replacement of CYA by TAC does not lead to a better management of severe SDNS. PMID- 16311258 TI - Severe hyperparathyroidism with bone abnormalities and metastatic calcification in rats with adenine-induced uraemia. AB - BACKGROUND: Marked parathyroid hyperplasia with bone diseases and vascular calcification are unsolved issues in dialysis patients. In this study, we made azotemic model rats by adenine feeding and analyzed the development and progression of the abnormalities. METHODS: Renal failure was induced in 8-week old male Wistar rats by feeding 0.75% adenine-containing diet for 6 weeks. Serum parameters, parathyroid hyperplasia, bone changes and metastatic calcification were examined at 2, 4 and 6 weeks. RESULTS: Progressive increase of serum creatinine and inorganic phosphate, and decreased levels of serum calcium and 1,25(OH)2D3 were confirmed. Markedly enlarged parathyroid glands and extremely high PTH levels were observed in all adenine-fed rats compared with the control (PTH: 199.3+/-58.0 vs 10.5+/-3.0 pmol/l, P<0.01, respectively, at 6 weeks). In cortical bone of the femur, the morphometric parameters showed increased bone resorption with increased fibrosis, whereas in the trabecular bone, bone resorption decreased and bone volume increased with a larger amount of osteoid compared with the control. Metastatic calcification in aorta, coronary artery and other soft tissues were also found in adenine-fed rats. CONCLUSIONS: Uraemic rats made by adenine diet developed severe abnormalities of calcium metabolism in a relatively short period and therefore they may serve as a useful model for the analysis of parathyroid hyperplasia and vascular calcification in chronic renal failure. PMID- 16311259 TI - Polyclonal activation of an IgA subclass against Staphylococcus aureus cell membrane antigen in post-methicillin-resistant S. aureus infection glomerulonephritis. PMID- 16311260 TI - Progression versus regression of chronic kidney disease. PMID- 16311261 TI - Obesity and hypertension--the issue is more complex than we thought. PMID- 16311262 TI - Renal cell carcinoma with an unusual presentation. PMID- 16311263 TI - Fluconazole dosing in continuous veno-venous haemofiltration (CVVHF): need for a high daily dose of 800 mg. AB - To cover intermediate sensitive Candida glabrata in ICU patients, fluconazole plasma peak levels at least in the range of 16-32 microg/ml appear necessary for treatment. Previous studies did not reach these fluconazole levels under continuous veno-venous haemofiltration (CVVHF) with dosages of 200-600 mg fluconzole daily. In the present study, nine patients simultaneously requiring CVVHF for treatment of acute oligoanuric renal failure and antimycotic therapy of Candida septicemia received fluconazole 800 mg/day. Fluconazole plasma levels were determined to evaluate whether this dosage is adequate to reach the advised fluconazole levels. Patients were dialysed on two consecutive days with an ultrafiltration rate (UF) of 1000 ml/h or 2000 ml/h, respectively, in a randomized order. The predilution was 800 ml/h and 1800 ml/h, respectively. The treatment was tolerated without adverse effects. All patients reached plasma fluconazole concentrations between 16 and 32 microg/ml, remaining in this range for a minimum of 1 up to 24 h with a mean of 9.6 h and a UF rate of 2000 ml/h, and 15.7 h with a UF rate of 1000 ml/h. So far, there are no in vivo data on the fluconazole plasma concentrations required for effective treatment. However, our data demonstrate, that at least the fluconazole concentrations desirable on the basis of in vitro susceptibility testing can be reached in critically ill patients on CVVHF in an ICU setting. However, in these patients, 800 mg fluconazole/day are necessary to achieve fungicidal drug concentrations. PMID- 16311264 TI - Haemodialysis vascular access problems in Canada: results from the Dialysis Outcomes and Practice Patterns Study (DOPPS II). AB - BACKGROUND: The optimal vascular access for chronic maintenance haemodialysis (HD) is the native arteriovenous fistula (AVF). Vascular access practice patterns are reported for a Canadian cohort of patients from the Dialysis Outcomes and Practice Patterns Study (DOPPS II). METHODS: DOPPS II is a prospective, observational study in 12 countries, including Canada. A representative random sample of 20 Canadian HD facilities and patients within those units were studied during 2002-2004. Canadian results were compared with those found in Europe and the USA. RESULTS: AVF use in Canadian prevalent (53%) and incident (26%) patients was lower than Canadian guidelines recommend (60%), and lower than in Europe [prevalent (74%), incident (50%)]. Despite 85% of Canadian HD patients having seen a nephrologist for > 1 month prior to starting dialysis, central venous catheter use in Canada (33% in prevalent patients, 70% in incident patients) was much higher than in Europe (prevalent 18%, incident 46%) and slightly higher than in the USA (prevalent 25%, incident 66%). This pattern is contrary to the preferences of Canadian medical directors and vascular access surgeons. The typical time from referral until permanent vascular access creation is substantially longer in Canada (61.7 days) than in Europe (29.4 days) or the USA (16 days). This longer delay time and higher catheter use in Canada may be a consequence of the significantly lower number of access surgeons per 100 HD patients in Canada (2.9) compared with the USA (8.1) and Europe (4.6). Furthermore, the median hours per week devoted to vascular access-related surgery per 100 patients is substantially lower in Canada (0.027 h) compared with the USA (0.082 h) and Europe (0.059 h). CONCLUSION: These findings suggest that Canadian chronic HD patients often rely on central venous catheters for vascular access, despite their known association with numerous detrimental outcomes in HD. Nephrologists, vascular access surgeons, interventional radiologists, other physicians and health care funding bodies must be more broadly educated about the priority of AVF creation as the preferred vascular access for chronic HD patients. They must work together to secure both the human and financial resources and other health care system enhancements to increase AVF creation rates in a timely manner. PMID- 16311265 TI - Up-regulation of hippocampal metabotropic glutamate receptor 5 in temporal lobe epilepsy patients. AB - Metabotropic glutamate receptors (mGluRs) are G protein-coupled receptors involved in the regulation of glutamatergic transmission. Recent studies indicate that excitatory group I mGluRs (mGluR1 and mGluR5) contribute to neurotoxicity and hyperexcitability during epileptogenesis. In this study, we examined the distribution of mGluR1alpha and mGluR5 immunoreactivity (IR) in hippocampal resection tissue from pharmaco-resistant temporal lobe epilepsy (TLE) patients. IR was detected with panels of receptor subtype specific antisera in hippocampi from TLE patients without (non-HS group) and with hippocampal sclerosis (HS group) and was compared with that of non-epileptic autopsy controls (control group). By immunohistochemistry and immunoblot analysis, we found a marked increase of mGluR5 IR in hippocampi from the non-HS compared with the control group. High mGluR5 IR was most prominent in the cell bodies and apical dendrites of hippocampal principal neurons and in the dentate gyrus molecular layer. In the HS group, this increase in neuronal mGluR5 IR was even more pronounced, but owing to neuronal loss the number of mGluR5-immunoreactive neurons was reduced compared with the non-HS group. IR for mGluR1alpha was found in the cell bodies of principal neurons in all hippocampal subfields and in stratum oriens and hilar interneurons. No difference in mGluR1alpha IR was observed between neurons in both TLE groups and the control group. However, owing to neuronal loss, the number of mGluR1alpha-positive neurons was markedly reduced in the HS group. The up-regulation of mGluR5 in surviving neurons is probably a consequence rather than a cause of the epileptic seizures and may contribute to the hyperexcitability of the hippocampus in pharmaco-resistant TLE patients. Thus, our data point to a prominent role of mGluR5 in human TLE and indicate mGluR5 signalling as potential target for new anti-epileptic drugs. PMID- 16311266 TI - Mitochondrial changes in skeletal muscle in amyotrophic lateral sclerosis and other neurogenic atrophies--a comment. PMID- 16311269 TI - Genetics of Parkinson's disease: LRRK2 on the rise. PMID- 16311270 TI - Autosomal dominant hereditary sensory neuropathy with chronic cough and gastro oesophageal reflux: clinical features in two families linked to chromosome 3p22 p24. AB - Autosomal dominant hereditary sensory neuropathy (HSN I) is a clinically and genetically heterogeneous group of disorders, and in some families it is due to mutations in the serine palmitoyltransferase (SPTLC1) gene. We have characterized two families with HSN I associated with cough and gastro-oesophageal reflux (GOR). From a large Australian family, 27 individuals and from a smaller family, 11 individuals provided clinical information and blood for genetic analysis. Affected individuals had an adult onset of paroxysmal cough, GOR and distal sensory loss. Cough could be triggered by noxious odours or by pressure in the external auditory canal (Arnold's ear-cough reflex). Other features included throat clearing, hoarse voice, cough syncope and sensorineural hearing loss. Neurophysiological and pathological studies demonstrated a sensory axonal neuropathy. Gastric emptying studies were normal, and autonomic function and sweat tests were either normal or showed distal hypohidrosis. Cough was likely to be due to a combination of denervation hypersensitivity of the upper airways and oesophagus, and prominent GOR. Most affected individuals were shown on 24 h ambulatory oesophageal pH monitoring to have multiple episodes of GOR, closely temporally associated with coughing. Hoarse voice was probably attributable to acid-induced laryngeal damage, and there was no evidence of vocal cord palsy. No other cause for cough was found on most respiratory or otorhinological studies. Linkage to chromosome 3p22-p24 has been found in both families, with no evidence of linkage to loci for known HSN I, autosomal dominant hereditary motor and sensory neuropathy, hereditary GOR or triple A syndrome. These families represent a genetically novel variant of HSN I, with a distinctive cough owing to involvement of the upper aerodigestive tract. PMID- 16311271 TI - Periventricular nodular heterotopia with overlying polymicrogyria. AB - Polymicrogyria (PMG) and periventricular nodular heterotopia (PNH) are two developmental brain malformations that have been described independently in multiple syndromes. Clinically, they present with epilepsy and developmental handicaps in both children and adults. Here we describe their occurrence together as the two major findings in a group of at least three cortical malformation syndromes. We identified 30 patients as having both PNH and PMG on brain imaging, reviewed clinical data and brain imaging studies (or neuropathology summary) for all, and performed mutation analysis of FLNA in nine patients. The group was divided into three subtypes based on brain imaging findings. The frontal perisylvian PNH-PMG subtype included eight patients (seven males and one female) between 2 days and 10 years of age. It was characterized by PNH lining the lateral body and frontal horns of the lateral ventricles and by PMG most severe in the posterior frontal and perisylvian areas, occasionally with extension to the parietal lobes beyond the immediate perisylvian cortex. The posterior PNH-PMG subtype consisted of 20 patients (15 male and 5 female) between 5 days and 40 years of age. It was characterized by PNH in the trigones, temporal and posterior horns of the lateral ventricles, and PMG most severe in the temporo-parieto occipital regions. The third type was found in 2 females aged 7 months and 2 years, and was characterized by severe congenital microcephaly and more diffuse cortical abnormality. The PNH-PMG subtypes described here have distinct imaging and clinical phenotypes that suggest multiple genetic aetiologies involving defects in multiple genes, and a shared pathophysiological mechanism for PNH and PMG. The frontal-perisylvian and posterior subtypes both had skewing of the sex ratio towards males, which suggests the possibility of X-linked inheritance. Delineation of these syndromes will also aid in providing more accurate diagnosis and prognostic information for patients with these malformations. PMID- 16311272 TI - Genetic variation in five Mediterranean populations of Juniperus phoenicea as revealed by inter-simple sequence repeat (ISSR) markers. AB - BACKGROUND AND AIMS: The assessment of the genetic variability and the identification of isolated populations within a given species represent important information to plan conservation strategies on a genetic basis. In this work, the genetic variability in five natural populations of Juniperus phoenicea, three from Sardinia, one from Cyprus and the last one in the Maritime Alps was analysed by means of ISSRs, on the hypothesis that the latter could have been a refugial one during the last glaciation. METHODS: ISSRs were chosen because of their ability to detect variation without any prior sequence information. The use of three primers yielded 45 reproducible, polymorphic bands, which were utilized to estimate the basic parameters of genetic variability and diversity. KEY RESULTS: All of the populations analysed harboured an adequate amount of genetic variability, with H(S) = 0.1299. The proportion of genetic diversity between populations has been estimated by G(ST) = 0.12. The three Sardinian populations are separated, as tested by AMOVA, from the Cyprus and the continental ones. CONCLUSIONS: The results indicate that geographical isolation has represented a major barrier to gene flow in Juniperus phoenicea. This work represents a first step towards a full genetic characterization of a conifer from the Mediterranean, a world biodiversity hotspot confronted with climate change, and thus contributes towards the planning of genetics-informed conservation strategies. PMID- 16311273 TI - Isolation and characterization of four ascorbate peroxidase cDNAs responsive to water deficit in cowpea leaves. AB - BACKGROUND AND AIMS: Abiotic stresses stimulate formation of active oxygen species in plant tissues. Among antioxidant mechanisms, H2O2 detoxication by ascorbate peroxidases (APX) plays an important role. Several APX isoforms exist in plant cells, and they have rarely been studied separately. The aim of this work was to study changes in cytosolic, peroxisomal, stromatic and thylakoid APX gene expression in response to progressive drought, rapid desiccation and application of exogenous abscisic acid in the leaves of cowpea (Vigna unguiculata) plants. METHODS: Two cowpea (V. unguiculata) cultivars, 'EPACE-1' which is drought-tolerant and '1183'which is drought-sensitive, were submitted to drought stress by withholding irrigation. Detached leaves were air-dried or treated with exogenous abscisic acid. APX cDNAs were isolated by PCR and cloned in plasmid vectors. Changes in gene expression were studied using reverse transcription PCR. KEY RESULTS: Four new V. unguiculata cDNAs encoding putative cytosolic, peroxisomal and chloroplastic (stromatic and thylakoidal) APX were isolated and characterized. In response to the different treatments, higher increases in steady-state transcript levels of the cytoplasmic and peroxisomal APX genes were observed in '1183' compared with 'EPACE-1'. On the other hand, the expression of the chloroplastic APX genes was stimulated earlier in the tolerant cultivar when submitted to progressive drought. CONCLUSIONS: Water deficit induced differences in transcript accumulation of APX genes between the two cultivars that were related to their respective tolerance to drought. Chloroplastic APX genes responded early to progressive water deficit in the tolerant plant, suggesting a capacity to efficiently detoxify active oxygen species at their production site. The more sensitive '1183' was also able to respond to drought by activating its whole set of APX genes. PMID- 16311274 TI - The presentation and survival of patients with non-cutaneous AIDS-associated Kaposi's sarcoma. AB - BACKGROUND: Acquired immune deficiency syndrome related Kaposi's sarcoma (AIDS KS) remains a significant cause of morbidity and mortality. We describe for the first time a proportion of patients with AIDS-KS who presented with no evidence of cutaneous disease. PATIENTS AND METHODS: From our cohort of 5932 individuals infected with the human immunodeficiency virus (HIV-1) treated in the HAART era, 319 were identified with KS. Of these, 11 patients (5.4%) were diagnosed with KS without the presence of any cutaneous disease. We compared their survival, clinical, immunological and virological characteristics to other individuals with KS. RESULTS: There were no statistically significant differences in survival, CD4 count or HIV viral load at KS presentation. We observed that tumour-associated oedema (P = 0.046) and non-oral gastrointestinal KS (P = 0.042) were significantly more common in patients with non-cutaneous KS. Only one case of non cutaneous KS was observed prior to the era of highly active anti-retroviral therapy (HAART). CONCLUSIONS: Non-cutaneous KS is a recognisable condition; patients should be treated with the standard of care as their prognosis is not inferior. This is likely to reflect a strong immune response, in the era of HAART. PMID- 16311275 TI - Phase III trial of 5-fluorouracil and leucovorin plus either 3H1 anti-idiotype monoclonal antibody or placebo in patients with advanced colorectal cancer. AB - BACKGROUND: The monoclonal antibody 3H1 mimics the external structure of the carcinoembryonic antigen (CEA). It therefore has the potential, via the anti idiotypic network, to stimulate immune responses to CEA that may benefit colorectal cancer patients. PATIENTS AND METHODS: A total of 630 patients with previously untreated metastatic colorectal cancer were randomised in a 2:1 fashion to receive bolus 5-fluorouracil (5-FU) and leucovorin (LV) plus either 3H1 (n = 422) or placebo (n = 208). RESULTS: The addition of 3H1 to 5-FU and LV did not result in increased toxicity. Survival for the full intent-to-treat population was 14.7 months for the 3H1 arm and 15.2 months for the placebo arm (P = 0.80). Anti-CEA antibody responses were observed in 70% of patients treated with 3H1. Patients with a negative CEA response had a median survival of 8.3 months (95% CI 7.5-11.0) compared with patients with a strong response: median survival not reached (P <0.001). CONCLUSION: 3H1 is safe and effectively induces immune responses to CEA. Addition of 3H1 to 5-FU and LV was not shown to improve overall patient outcomes. However, improved survival in patients developing anti CEA responses to 3H1 are provocative and should be studied in further clinical trials. PMID- 16311276 TI - Bedside confirmation of a persistent left superior vena cava based on aberrantly positioned central venous catheter on chest radiograph. AB - This is a report of incidental diagnosis of a persistent left superior vena cava (PLSVC) based on an abnormal positioning of central venous catheter seen on chest radiograph and an abnormal pressure waveform. Non-invasive bedside tests included venography with simultaneous chest radiograph and a transthoracic echocardiography with an agitated saline microbubble contrast. These tests led to the diagnosis of PLSVC. Although PLSVC is the most common venous thoracic anomaly that produces a diagnostic dilemma, not many anaesthetists and intensivists are familiar with its appearance, diagnosis and implications. The clinical significance of PLSVC and diagnostic options are discussed. PMID- 16311277 TI - Effects of three different dose regimens of magnesium on propofol requirements, haemodynamic variables and postoperative pain relief in gynaecological surgery. AB - BACKGROUND: In this double-blind, randomized, placebo-controlled study we compared the effects of three different dose regimens of magnesium on intraoperative propofol and atracurium requirements, and postoperative morphine consumption in patients undergoing gynaecological surgery. METHODS: Eighty women were allocated to four equal groups. The control group received normal saline; magnesium groups received 40 mg kg(-1) of magnesium before induction of anaesthesia, followed by i.v. infusion of normal saline, magnesium 10 mg kg(-1) h(-1) or magnesium 20 mg kg(-1) h(-1) for the next 4 h. Propofol infusion was targeted to keep bispectral index values between 45 and 55. Postoperative analgesia was achieved using PCA with morphine. RESULTS: Magnesium groups required significantly less propofol [mean (sd) 121.5 (13.3), 102.2 (8.0) and 101.3 (9.7) microg kg(-1) min(-1) respectively] than the control group (140.7 (16.5) microg kg(-1) min(-1)). Atracurium use was significantly higher in the control group than magnesium groups [0.4 (0.06) vs 0.34 (0.06), 0.35 (0.04), 0.34 (0.06) mg kg(-1) h(-1) respectively]. Morphine consumption was significantly higher in control group than magnesium groups on the first postoperative day [0.88 (0.14) vs 0.73 (0.17), 0.59 (0.23), 0.53 (0.21) mg kg(-1) respectively]. The heart rate was lower in magnesium groups and 20 mg kg(-1) h(-1) infusion group demonstrated the lowest values. CONCLUSION: Magnesium 40 mg kg(-1) bolus followed by 10 mg kg(-1) h(-1) infusion leads to significant reductions in intraoperative propofol, atracurium and postoperative morphine consumption. Increasing magnesium dosage did not offer any advantages, but induced haemodynamic consequences. PMID- 16311278 TI - Bougie-guided insertion of the ProSeal laryngeal mask airway has higher first attempt success rate than the digital technique in children. AB - BACKGROUND: We tested the hypothesis that bougie-guided insertion of the ProSeal laryngeal mask airway (ProSeal LMA) has higher success rate than the digital technique in children. METHODS: One hundred and twenty children (ASA I-II, aged 1 16 yr) were randomly allocated for ProSeal LMA insertion using the digital or bougie-guided technique. The digital technique was performed according to the manufacturer's instructions. The bougie-guided technique involved priming the drain tube with a bougie, placing the bougie in the oesophagus under direct vision and railroading the ProSeal LMA into position. Unblinded data were collected about ease of insertion (number of attempts and time taken to provide an effective airway), efficacy of seal, ease of gastric tube placement, haemodynamic responses and blood staining. Blinded data were collected about postoperative airway morbidity. RESULTS: The first attempt success rate was higher for the bougie-guided technique (59/60 vs 52/60, P=0.015), but effective airway time was longer (37 vs 32 s, P<0.001). There were no differences in efficacy of seal, ease of gastric tube placement, haemodynamic responses, blood staining or postoperative airway morbidity. CONCLUSION: We conclude that bougie guided insertion of the ProSeal LMA has a higher first attempt success rate than the digital technique in children. PMID- 16311279 TI - Effect of fluid loading with saline or colloids on pulmonary permeability, oedema and lung injury score after cardiac and major vascular surgery. AB - BACKGROUND: The optimal type of fluid for treating hypovolaemia without evoking pulmonary oedema is still unclear, particularly in the presence of pulmonary vascular injury, as may occur after cardiac and major vascular surgery. METHODS: In a single-centre, prospective, single-blinded clinical trial 67 mechanically ventilated patients were randomly assigned to receive saline, gelatin 4%, HES 6% or albumin 5%, according to a 90 min fluid loading protocol with target central venous pressure of 13 and pulmonary capillary wedge pressure of 15 mm Hg, within 3 h after cardiac or major vascular surgery. Before and after the protocol, we recorded haemodynamics and ventilatory variables and took chest radiographs. The pulmonary vascular injury was evaluated using the 67Ga-transferrin pulmonary leak index (PLI) and extravascular lung water (EVLW). Plasma colloid osmotic pressure (COP) was determined and the lung injury score (LIS) was calculated. RESULTS: More saline was infused than colloid solutions (P<0.005). The COP increased in the colloid groups and decreased in patients receiving saline. Cardiac output increased more in the colloid groups. At baseline, PLI and EVLW were above normal in 60 and 30% of the patients, with no changes after fluid loading, except for a greater PLI decrease in HES than in gelatin-loaded patients. The oxygenation ratio improved in all groups. In the colloid groups, the LIS increased, because of a decrease in total respiratory compliance, probably associated with an increase in intrathoracic plasma volume. CONCLUSIONS: Provided that fluid overloading is prevented, the type of fluid used for volume loading does not affect pulmonary permeability and oedema, in patients with acute lung injury after cardiac or major vascular surgery, except for HES that may ameliorate increased permeability. During fluid loading, changes in LIS (and respiratory compliance) do not represent changes in pulmonary permeability or oedema. PMID- 16311280 TI - Obese parturients have lower epidural local anaesthetic requirements for analgesia in labour. AB - BACKGROUND: There are no studies comparing local anaesthetic requirements for obese and normal parturients. Obesity has been associated with a higher incidence of Caesarean section and higher levels of epidural block have also been found in obese obstetric patients, suggesting they may require less local anaesthetic. The aim of our study was to estimate the minimum local analgesic concentration (MLAC) of bupivacaine for obese and non-obese parturients. METHODS: Otherwise healthy parturients (n=32) requesting epidural analgesia were enrolled in this up-down sequential allocation study. Women were in active labour (3-6 cm cervical dilatation) with visual analogue pain scores (VAPS) >40/100 mm. Subjects with BMI >30 kg m(-2) were allocated to the obese group and BMI < or = 30 kg m(-2) were allocated to the normal group. The initial epidural dose for both groups was 20 ml 0.1% w/v bupivacaine (20 mg), with a dosing increment of 0.01% w/v VAPS < or = 10/100 mm defined effective analgesia. The MLAC was estimated using up-down reversals and probit regression with P<0.05 as significant. RESULTS: Groups were similar except for BMI and weight (P<0.001). Local anaesthetic requirements were significantly (P<0.001) reduced by a factor of 1.68 (95% CI 1.32-2.29) in the obese group, with significantly higher initial level of block (P<0.001). CONCLUSION: We found obese parturients to have significantly decreased epidural bupivacaine analgesic requirements. A contributing factor to obese patients having more difficult labours may be that relatively larger doses of local anaesthetic are administered than actually required. It may be worth considering lowering the concentrations and doses with which we initiate analgesia in obese parturients. PMID- 16311281 TI - Arytenoid cartilage dislocation caused by a double-lumen endobronchial tube. AB - Following surgery requiring the use of a double-lumen endobronchial tube, a patient immediately complained of persistent severe hoarseness. On the third day after the operation, fibreoptic laryngoscopy revealed posterolateral dislocation of the left arytenoid cartilage. By the sixth day of the operation, a slight improvement was observed in the hoarseness without treatment and a spontaneous recovery of arytenoid cartilage dislocation was expected. The patient did not consent to surgical treatment, and therefore a conservative therapy was selected. Ten weeks after the operation, it was found that the dislocated left arytenoid cartilage had spontaneously repositioned and the patient regained his normal voice. The causes and treatment options are discussed. PMID- 16311282 TI - Influence of improved teaching on medical students' acquisition and retention of drug administration skills. AB - BACKGROUND: Drug administration error is a major problem causing substantial morbidity and mortality worldwide. Lack of education about drug administration appears to be a causative factor. We devised an online teaching module for medical students and assessed its short- and long-term efficacy. METHODS: One hundred and thirty clinical medical students were invited to undertake additional, online, teaching about drug administration. Those participating were identified and the number of web pages viewed recorded. The students' knowledge retention was tested by means of drug administration questions incorporated into routine assessments and examinations over the next 6 months. Other indices of all students' performance were recorded to correct for confounding factors. RESULTS: Just over half (52%) responded to the invitation to participate. The amount of interest they showed in the teaching module correlated positively with their performance in questions about drug administration, although the latter waned over time. Surprisingly, correcting for students' general ability and keenness revealed that the less able students were most likely to undertake the teaching module. CONCLUSIONS: Additional online teaching about drug administration improves students' knowledge of the topic but clearly requires reinforcement; however, only about half the students took up the option. Medical students must acquire these fundamental skills, and online teaching can help. Medical educators must ensure that drug administration is taught formally to all students as part of the curriculum and must understand that it may require additional teaching. PMID- 16311283 TI - Incidence of adrenal insufficiency after severe traumatic brain injury varies according to definition used: clinical implications. AB - BACKGROUND: Adrenal insufficiency impacts on the haemodynamic management of patients in intensive care. Very little is known about the incidence of adrenal insufficiency in the first 10 days after traumatic brain injury. METHODS: We retrospectively reviewed the charts of 113 traumatic brain injury patients within 10 days of their injury. They all had a high-dose corticotropin stimulation test performed because of haemodynamic instability. Blood cortisol concentrations were measured at baseline, 30 and 60 min after the administration of high-dose corticotropin. The incidence of adrenal insufficiency was determined according to various definitions used in the literature. RESULTS: The baseline cortisol concentration was <414 nmol litre(-1) (15 microg dl(-1)) in 78% of patients and <690 nmol litre(-1) (25 microg dl(-1)) in all patients. The cortisol concentration did not rise above 500 nmol litre(-1) (18 microg dl(-1)) at 30 and 60 min in 49 and 22% of patients, respectively. The cortisol concentration did not rise by 250 nmol litre(-1) (9 microg dl(-1)) at 30 and 60 min in 48 and 25% of patients respectively. Primary adrenal insufficiency defined by an abnormal baseline cortisol concentration and an abnormal response to the high-dose corticotropin stimulation test was present in 13-28% of patients according to the cut-off values used. CONCLUSIONS: The incidence of adrenal insufficiency varies from 25 to 100% in the first 10 days after traumatic brain injury. The range of incidences reported illustrates the need for standardization of the definition of adrenal insufficiency. This has a direct impact on treatment. Sampling at 60 min after the high-dose corticotropin stimulation test seems to correlate better with the maximum secreting capacity of the adrenal glands. PMID- 16311284 TI - Physicians' perceptions and attitudes regarding inappropriate admissions and resource allocation in the intensive care setting. AB - BACKGROUND: Physicians' perceptions regarding intensive care unit (ICU) resource allocation and the problem of inappropriate admissions are unknown. METHODS: We carried out an anonymous, self-administered questionnaire survey to assess the perceptions and attitudes of ICU physicians at all 20 ICUs in Milan, Italy, regarding inappropriate admissions and resource allocation. RESULTS: Eighty-seven percent (225/259) of physicians responded. Inappropriate admissions were acknowledged by 86% of respondents. The reasons given were clinical doubt (33%); limited decision time (32%); assessment error (25%); pressure from superiors (13%), referring clinician (11%) or family (5%); threat of legal action (5%); and an economically advantageous 'Diagnosis Related Group' (1%). Respondents reported being pressurized to make more 'productive' use of ICU beds by Unit heads (frequently 16%), hospital management (frequently 10%) and colleagues (frequently 4%). Five percent reported refusing appropriate admissions following 'indications' not to admit financially disadvantageous cases. Admissions after elective surgery prioritized patients from profitable surgical departments: frequently for 6% of respondents and occasionally for 15%. Sixty-seven percent said they frequently received requests for appropriate admissions when no beds were available. This was considered sufficient reason to withdraw treatment from patients with lower survival probability (sometimes 21%) or for whom nothing more could be done (sometimes 51%, frequently 11%). CONCLUSIONS: Inappropriate ICU admissions were perceived as a common event but were mainly attributed to difficulties in assessing suitability. Physicians were aware that their decisions were often influenced by factors other than medical necessity. Economic influences were perceived as limited but not negligible. Decisions to forgo treatment could be influenced by the need to admit other patients. PMID- 16311285 TI - Factors influencing emergency intubation in the pre-hospital setting--a multicentre study in the German Helicopter Emergency Medical Service. AB - BACKGROUND: Definitive airway control by endotracheal intubation (ETI) is standard of care in pre-hospital airway management. However, there are specific factors that may influence and complicate ETI. METHODS: Prospective, descriptive study at three German Helicopter Emergency Medical Services (HEMS) over a 1-yr period. We examined the success and complication rate for field intubation performed by trauma anaesthetists. RESULTS: In 342 patients (9.3%) ETI was performed. The overall success rate was 100%; in 87.4% the first attempt was successful, whereas in 11.1% a second and in 1.5% a third ETI attempt was necessary. No patient required a surgical intervention. Limited access to the patient was found upon arrival at the scene in 20.2% of the patients and in 9.6% of the patients at the time of ETI attempt. An orotracheal ETI technique was used in all patients. In the patients in whom only one ETI attempt was necessary for successful intubation, the assessment of ETI conditions was rated 'very good' or 'good' in 94.7%, but in those requiring a second or third ETI attempt this was reduced to 68.6 and 20.0%, respectively. Difficulties encountered during ETI included blood (19.9%), vomit/debris (15.8%) and secretions (13.8%) in the upper airway; anatomical reasons (11.7%), patient position (9.6%) and surrounding conditions (9.1%), making laryngoscopy more difficult. CONCLUSIONS: Despite various factors increasing the difficulties in managing the airway in the field, definitive airway control by ETI seems to be safe practice. PMID- 16311286 TI - Equivalent dose of ephedrine and phenylephrine in the prevention of post-spinal hypotension in Caesarean section. AB - BACKGROUND: Comparative studies of ephedrine and phenylephrine in prevention of hypotension after spinal anaesthesia for Caesarean section have lacked a consensus on dose equivalence. The aim of this study was to determine the minimum vasopressor dose for each of these drugs to calculate the dose ratio for clinical equivalence in the prevention of hypotension. METHODS: Patients with a normal singleton pregnancy beyond 36 weeks gestation undergoing elective Caesarean section under spinal anaesthesia were randomized into two groups. The first patient in Group A received 50 mg of ephedrine in saline 0.9% w/v, 500 ml, at 999 ml h(-1), the maximum rate possible on the pump and the first patient in Group B received 500 microg of phenylephrine in saline 0.9% w/v, 500 ml, at the same rate. The initial dose for dilution was an arbitrary choice. The dose of vasopressor in the saline bag for every subsequent patient was established by the efficacy of the dose in preventing hypotension in the previous patient according to the technique of up-down sequential allocation. Minimum vasopressor dose for each drug was determined according to the Dixon-Massey formula. RESULTS: The minimum vasopressor dose in saline 500 ml was 532.9 microg (95% CI 506.0-559.8) for phenylephrine and 43.3 mg (95% CI 39.2-47.3) for ephedrine. The concentration needed for equivalence at an infusion rate of 999 ml h(-1) was 1.07 microg ml(-1) for phenylephrine and 86.66 microg ml(-1) for ephedrine. Mean (sd) dose used for phenylephrine was 496.45 (78.3) microg and for ephedrine 39.64 (6.33) mg. CONCLUSION: This study demonstrates a potency ratio of 81.2 (95% CI 73.0-89.7) for equivalence between phenylephrine and ephedrine in prevention of hypotension after spinal anaesthesia for Caesarean section. PMID- 16311287 TI - Strategies and clinical outcome of 250 cycles of Preimplantation Genetic Diagnosis for single gene disorders. AB - BACKGROUND: We report on our experience with preimplantation genetic diagnosis (PGD) for single gene disorders (SGDs), from 1999 to 2004, describing strategies and overall clinical outcome of 250 cycles in 174 couples for 23 different genetic conditions. METHODS: PGD cycles included 15 for autosomal dominant, 148 for autosomal recessive and 19 for X-linked SGDs. In addition, 68 cycles of PGD for SGDs were performed in combination with HLA matching. The strategy in each case used an initial multiplex PCR, followed by minisequencing to identify the mutation(s) combined with multiplex PCR for closely linked informative markers to increase accuracy. Linkage analysis, using intragenic and/or extragenic polymorphic microsatellite markers, was performed in cases where the disease causing mutation(s) was unknown or undetectable. RESULTS: In 250 PGD cycles, a total of 1961 cleavage stage embryos were biopsied. PCR was successful in 3409 out of 3149 (92.4%) biopsied blastomeres and a diagnosis was possible in 1849 (94.3%) embryos. Four hundred and twenty-seven embryos were transferred in 211 cycles, resulting in 71 pregnancies (33.6% per embryo transfer), including 15 biochemical pregnancies, six spontaneous miscarriages, two ectopic pregnancies, which were terminated, and nine pregnancies which are still ongoing. The remaining pregnancies were confirmed to be unaffected and went to term without complications, resulting in the birth of 35 healthy babies. CONCLUSIONS: Minisequencing for mutation detection combined with multiplex fluorescence PCR for linkage analysis is an efficient, accurate and widely applicable strategy for PGD of SGDs. Our experience provides a further demonstration that PGD is an effective clinical tool and a useful option for many couples with a high risk of transmitting a genetic disease. PMID- 16311288 TI - Etoposide induces chromosomal abnormalities in mouse spermatocytes and stem cell spermatogonia. AB - BACKGROUND: Etoposide (ET) is a chemotherapeutic agent widely used in the treatment of leukaemia, lymphomas and many solid tumours such as testicular and ovarian cancers, all of which are common in patients of reproductive age. The purpose of the study was to characterize the long-term effects of ET on male germ cells using sperm fluorescence in situ hybridization (FISH) analyses. METHODS: Chromosomal aberrations (partial duplications and deletions) and whole chromosomal aneuploidies were detected in sperm of mice treated with a clinical dose of ET. Semen samples were collected at 25 and 49 days after dosing to investigate the effects of ET on meiotic pachytene cells and spermatogonial stem cells, respectively. RESULTS: ET treatment resulted in major increases in the frequencies of sperm-carrying chromosomal aberrations in both meiotic pachytene (27- to 578-fold) and spermatogonial stem-cells (8- to 16-fold), but aneuploid sperm were induced only after treatment of meiotic cells (27-fold) with no persistent effects in stem cells. CONCLUSION: These results show that ET may have long-lasting effects on the frequencies of sperm with structural aberrations. This has important implications for cancer patients undergoing chemotherapy with ET because they may remain at higher risk for abnormal reproductive outcomes long after the end of chemotherapy. PMID- 16311289 TI - Spermatogonial survival after grafting human testicular tissue to immunodeficient mice. AB - BACKGROUND: The xenografting of pre-pubertal human testicular tissue to an immunodeficient mouse is a theoretical strategy for restoring fertility in childhood cancer patients, while circumventing the risk of malignant recurrence. This study aimed at comparing the grafting of pre-pubertal and adult murine testicular tissue, as well as that of human adult testicular tissue, to two immunodeficient recipients, i.e. Swiss Nude mice and SCID-NOD mice. MATERIALS AND METHODS: In this study, we evaluated the survival of pre-pubertal and adult murine testicular tissues, and that of adult human testicular tissue after subcutaneous grafting to immunodeficient mice. RESULTS: After allografting pre pubertal testicular tissue pieces, meiotic cells were observed in 69.1% of the grafts, while complete spermatogenesis was observed in 30.9%. All grafts of adult murine testicular tissue and 59.5% of the adult human testicular grafts showed sclerosis. However, in 21.6% of the adult human testicular grafts, spermatogonia were still observed, with increasing sclerosis in time. No significant differences were observed between the two mouse models under evaluation. CONCLUSION: After xenografting human adult testicular tissue to a recipient mouse, spermatogonia were maintained over a period of >195 days. However, in order to prove xenografting as a method for external germ line storage, the transplants should have a more immature developmental stage. Moreover, not only the developmental status of the tissue at the time-point of grafting, but also the structural organisation of the seminiferous epithelium, might influence the development of the testicular tissue. PMID- 16311290 TI - HLA-G is expressed by the glandular epithelium of peritoneal endometriosis but not in eutopic endometrium. AB - BACKGROUND: HLA-G is a major histocompatability antigen with documented immune regulatory function. Various epithelial cancers and tissue allografts have been noted to express HLA-G, which is postulated to aid in their escape from immunosurveillance. We evaluated peritoneal endometriosis and eutopic endometrium for the expression of HLA-G protein and gene transcript. METHODS: Two experiments were performed: (i) archived tissue blocks from peritoneal endometriotic lesions (n = 15) and eutopic endometrium (n = 12) were evaluated for extent of protein immunostaining, and (ii) eutopic endometrial biopsies from women without (n = 17) and with (n = 24) endometriosis, and peritoneal endometriotic lesions (n = 14) were evaluated for presence of RNA transcript by in situ hybridization. RESULTS: HLA-G protein localized in the glandular epithelium of 14 of 15 (93.3%) peritoneal endometriotic lesions, but not in stromal cells. HLA-G protein staining was absent in endometrial biopsies (n = 12). HLA-G gene transcript localized to the glandular epithelium in 13 of 14 (92.8%) peritoneal endometriotic lesions. HLA-G transcript was never observed in eutopic endometrium, regardless of cycle stage or whether from women with (n = 24) or without (n = 18) endometriosis. CONCLUSIONS: HLA-G is expressed by endometriotic glandular epithelium but not by eutopic endometrium under normal conditions. Differential expression of HLA-G suggests that peritoneal inflammation or cellular stress may up-regulate mechanisms to promote ectopic endometrial survival. PMID- 16311291 TI - Activin receptor expression and induction of apoptosis in rat blastocysts in vitro. AB - BACKGROUND: Apoptosis, a process of normal embryonic development, is enhanced in blastocyst from diabetic rats. Nevertheless, glucose seems not to be the only factor involved. Activin A, a TGF-beta family member, is also increased in maternal serum from diabetic pregnancy. METHODS: Flushing medium, blastocysts and uterine cells were obtained from 5 day old pregnant rats. The presence of activin A in flushing medium was investigated by western blotting. RT-PCR was used to test for the presence of activin betaA subunit mRNA in cultured uterine cells. Blastocysts were stained by immunohistochemistry for activin receptor types IIA and IIB, and chromatin degradation (apoptosis) was investigated by terminal transferase-mediated dUTP nick end labelling in blastocysts exposed in vitro to activin. RESULTS: In this study, we demonstrate the presence of activin A protein in fluid from rat uterine horns at day 5 of pregnancy, as well as the presence of activin A receptors type IIB in the trophectoderm and inner cell mass and activin A receptor type IIA in trophectoderm cells only. Activin A increases the chromatin degradation level in vitro. CONCLUSIONS: Activin A protein was found in fluid from uterine horns, and mRNA expression of betaA activin subunit in cultured uterine cells suggests probable secretion from decidual cells. Moreover, activin A increases specifically the apoptosis level in rat blastocyst in vitro. PMID- 16311292 TI - Value of the sperm chromatin dispersion test in predicting pregnancy outcome in intrauterine insemination: a blind prospective study. AB - BACKGROUND: Sperm DNA integrity has been used as a new marker of sperm quality in the prediction of pregnancy. Nevertheless, no previous study has been performed by analysing the same samples that were employed in assisted reproduction. The main objective of this work was to correlate sperm chromatin dispersion (SCD), measured by the SCD test, with semen parameters and pregnancy outcome in intrauterine insemination (IUI). METHODS: A total of 100 semen samples obtained from males of couples undergoing IUI were analysed by the SCD test before and after swim-up, and the results were correlated with semen parameters and pregnancy outcome. RESULTS: SCD was negatively correlated with sperm motility in both ejaculated and processed semen. Sperm recovered by swim-up did not show a significant improvement in DNA integrity. No correlation was found between SCD and pregnancy outcome in IUI. CONCLUSIONS: DNA dispersion, as measured by the SCD test, is not correlated with pregnancy outcome in IUI. PMID- 16311293 TI - Aortic function is compromised in a rat model of polycystic ovary syndrome. AB - BACKGROUND: Arterial mechanical parameters are modified in women with polycystic ovary syndrome (PCOS), before and during pregnancy. This study tested the hypothesis that aortic mechanics and endothelial function are modified in the mifepristone-treated rat model of PCOS. METHODS: Female rats injected daily with mifepristone or vehicle for 7-9 days were assessed by ultrasound to allow estimation of aortic stiffness index and compliance. The influence of acetylcholine (ACh) and sodium nitroprusside (SNP) on dissected phenylephrine contracted aortic rings was assessed. RESULTS: Aortic compliance was reduced by 67% in mifepristone-treated rats versus controls (P<0.05), while stiffness index was increased 2.3-fold (P<0.02). ACh-induced dilation was less in aortic rings from mifepristone-treated rats (P=0.022) and was less sensitive to the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) (P<0.001), while SNP-induced dilation was greater (P=0.001). CONCLUSIONS: Aortic mechanics in vivo and endothelial function in vitro were consistently perturbed in mifepristone-treated rats. Aortic ring behaviour suggested that NO release was depressed or degradation elevated, with a compensatory increase in NO sensitivity and/or activation of a non-NO-mediated relaxation mechanism. The mifepristone treated rat is a valid model for investigation of the vascular deficits seen in PCOS. PMID- 16311294 TI - Oocyte donation in patients with Turner's syndrome: a successful technique but with an accompanying high risk of hypertensive disorders during pregnancy. AB - BACKGROUND: Few data are available on pregnancy rate and obstetrical outcome after oocyte donation in Turner's syndrome patients. We conducted a retrospective analysis on the outcome of this subgroup. METHODS: Thirty oocyte donation cycles with fresh embryo transfer were performed in 21 patients between 2001 and 2004. RESULTS: The mean (+/-SD) age of the recipients was 33.1+/-1.8 years. The median (range) number of transferred embryos per cycle was two (1-4). Seventeen pregnancies were obtained (57%), of which 12 were clinical (40%). The implantation rate and the ongoing pregnancy rate were 22% (15 out of 68) and 30% (nine out of 30), respectively. Premature delivery was observed in 50% (four out of eight) of the pregnancies and intrauterine growth retardation in 55.5% (five out of nine) of the fetuses. Hypertensive disorders occurred in five out of eight pregnancies (three pre-eclampsias). CONCLUSIONS: Turner's syndrome patients achieve acceptable pregnancy rates after oocyte donation. A high rate of pregnancy-associated hypertensive disorders was observed which have led to a high rate of prematurity and intrauterine growth restriction. Although the number of cases in this study is limited, these results call for the need for intensive surveillance of such pregnancies. In order to reduce the risk of hypertensive disorders induced by multiple pregnancies, single embryo transfer should be proposed. PMID- 16311295 TI - The psychological impact of mild ovarian stimulation combined with single embryo transfer compared with conventional IVF. AB - BACKGROUND: The objective of this study was to assess the psychological implications of mild ovarian stimulation combined with single embryo transfer (SET) during a first IVF cycle. METHODS: We conducted a randomized controlled two centre trial. Three hundred and ninety-one couples were randomized to undergo either mild ovarian stimulation with GnRH antagonist co-treatment and SET (n=199) or conventional GnRH agonist long protocol ovarian stimulation with double embryo transfer (DET) (n=192). Women completed the Hospital Anxiety and Depression Scale, the Hopkins Symptom Checklist and the Subjective Sleep Quality Scale at baseline, on the first day of ovarian stimulation and following embryo transfer. Affect was assessed daily with the Daily Record Keeping Chart from the first day of ovarian stimulation until the day treatment outcome became known. RESULTS: The conventional IVF group experienced elevated levels of physical and depressive symptoms during pituitary downregulation. At oocyte retrieval, this group experienced more positive affect and less negative affect than the mild IVF group. In the conventional IVF group, cycle cancellation was associated with less positive and more negative affect. CONCLUSIONS: During the first IVF treatment cycle, mild ovarian stimulation and SET does not lead to more psychological complaints than conventional IVF. PMID- 16311296 TI - Nicotine does not affect vascularization but inhibits growth of freely transplanted ovarian follicles by inducing granulosa cell apoptosis. AB - BACKGROUND: There is clear support for an association between smoking and decreased female fecundity and fertility. Cigarette smoke appears to have adverse effects along a continuum of reproductive processes. We therefore studied the effect of nicotine on follicular growth and vascularization of freely transplanted ovarian follicles. METHODS: We used the skinfold chamber model in Syrian golden hamsters, which allows the in vivo microscopy of follicular grafts. Animals were treated daily with nicotine at doses mimicking low-rate and high rate smokers (0.2 and 1.0 mg/kg body weight subcutaneously). Saline-treated animals served as controls. To further evaluate the effect of nicotine on angiogenesis, an in vitro aortic ring assay was used. RESULTS: The re vascularisation rate of follicles was similar in nicotine-treated animals and controls. During the 7 days after transplantation, nicotine further caused a dose dependent inhibition of follicular growth. In contrast, the vascularized area and microvessel density were not affected by the nicotine exposure. In vitro aortic ring assays confirmed that nicotine does not influence sprouting and microvessel formation. However, immunohistochemistry for cleaved caspase-3 revealed a large extent of granulosa cell apoptosis within transplanted follicles of high-dose nicotine-treated animals. CONCLUSIONS: Nicotine as one toxic component of cigarette smoke does not affect vascularization, but adversely influences follicular growth by an increase in apoptotic cell death. As follicular growth is a crucial step in normal ovulation and fertilization, nicotine-induced cell apoptosis may represent one of the mechanisms underlying the well-established link between smoking and fertility disorders. PMID- 16311297 TI - Antenatal care in singleton pregnancies after ICSI as compared to spontaneous conception: data from a prospective controlled cohort study in Germany. AB - BACKGROUND: The aim was to compare the antenatal care and the intake of vitamins and medications of infertility patients with singleton pregnancies after ICSI to women with a spontaneously conceived singleton pregnancy. METHODS: The data on the antenatal care of 2055 singleton pregnancies after ICSI from a prospective controlled multicentre study in Germany were analysed. The prospectively collected data of the control group of 7861 singletons were retrospectively assessed for the present analysis. RESULTS: The ICSI patients were significantly older (32.9 versus 27.0 years, P<0.019) and more likely to be obese (body mass index>or=30 kg/m2: 13.9 versus 4.8%, P<0.001) than the controls. The control mothers were significantly more likely to smoke (19.2%) or to consume alcohol (23.5%) during pregnancy than the ICSI mothers (7.4 and 0.6% respectively). Only 38.1% of ICSI patients took folic acid before conception. Only 61.7% of ICSI patients received an iodine supplementation at some point during pregnancy. ICSI patients went more regularly to the routine antenatal care consultations at the gynaecologist and had ultrasound examinations performed more regularly than the controls. The absolute number of ultrasound examinations was significantly higher in the ICSI group (13.6+/-6.0 versus 4.1+/-2.4). CONCLUSION: In spite of the intensive use of antenatal medical care by patients pregnant after ICSI, simple methods, such as the use of supplementary iodine and periconceptional folic acid, were used rarely. This indicates that counselling pregnant patients after ICSI about periconceptional and antenatal care seems to be insufficient in Germany and must be improved. PMID- 16311298 TI - Defining the rise of serum HCG in viable pregnancies achieved through use of IVF. AB - BACKGROUND: We aimed to characterize the rate of HCG rise associated with viable IVF pregnancies, and to evaluate the association between HCG rise and potentially influential factors. METHODS: We performed a retrospective cohort analysis of all viable pregnancies achieved through IVF at two centres between January 1999 and March 2004. RESULTS: Of the 455 pregnancies resulting in live births, 391 met inclusion criteria and contributed a total of 1052 HCG values. Using random effects models, the best pattern to describe the rise of log HCG was quadratic with the rate of increase slowing at 24 days post-oocyte retrieval. Limiting the analysis to measurements below the discriminatory zone, the linear model adequately characterized the profile. The average slope was 0.403, yielding a predicted increase of 1.50 (50% increase) in 1 day and 2.24 (124%) in 2 days. In the final model, absolute HCG values, but not rate of rise, were significantly higher for twins and triplets and significantly lower for patients with BMI>30 kg/m2. CONCLUSIONS: The HCG profile of viable pregnancies conceived with IVF is quadratic with an earlier plateau than has been reported for non-IVF pregnancies. The average rate of rise is comparable to previous estimates in symptomatic spontaneous conceptions. PMID- 16311299 TI - Fatty acid metabolism in human preimplantation embryos. AB - BACKGROUND: Little is known of fatty acid metabolism in human embryos. This information would be useful in developing metabolic tests of embryo quality and improving embryo culture media. METHODS: The fatty acid composition of human embryos and their ability to accumulate 13C labelled fatty acids was assessed in relation to the stage of development using gas-chromatography and combustion isotope-ratio-mass spectrometry. RESULTS: Compared with embryos which did not develop beyond the 4-cell stage, those that did had significantly higher concentrations of the unsaturates, linoleic (12% versus 3%; P=0.02) and oleic (14% versus 7%; P=0.02), and a lower concentration of total saturates (62% versus 77%; P=0.04). There was uptake of both 13C linoleic and palmitic, but the developmental pattern was different for each fatty acid. The net accumulation in pmol/embryo/24h for palmitic was 1 at the 2-cell to <8-cell stage, 4 at the 8 cell-morula stage and negligible at the blastocyst stage. For linoleic, there was little net accumulation at the 2-cell to <8-cell stage, 8 (8-cell-morula stage) and 17 pmol/embryo/24 h (blastocyst stage). CONCLUSION: Preimplantation human embryos actively take up individual fatty acids at different rates at different stages of development. The high unsaturated concentration at the later stages of development may be explained by preferential uptake of linoleic acid. PMID- 16311300 TI - Effects of mifepristone on proliferation and apoptosis of human endometrium in new users of medroxyprogesterone acetate. AB - BACKGROUND: Mifepristone has been demonstrated to decrease breakthrough bleeding (BTB) in users of progestin-only contraceptives. METHODS: Endometrial biopsies were collected from 50 normal cycling women who were new users of depot medroxyprogesterone acetate (DMPA) randomized to receive either mifepristone or placebo before, during and after treatment. Proliferation, apoptosis and sex steroid receptors were evaluated by either immunohistochemistry or TUNEL assay. RESULTS: Administration of mifepristone to DMPA-exposed endometrium for 1 week significantly increased endometrial expression of Ki-67 (MKI67), estrogen receptor (ER)alpha and progesterone receptors A and B (PRAB) and decreased the number of TUNEL-positive and caspase-3 (CASP3)-active cells in the endometrial stroma. However, after 10 weeks of mifepristone treatment, no significant difference in proliferation, apoptosis and the expression of ERalpha or PRAB could be detected between the endometrium treated with DMPA alone and endometrium treated with mifepristone and DMPA. CONCLUSIONS: Administration of mifepristone to DMPA users significantly increases endometrial proliferation and decreases endometrial stromal apoptosis in the short term. Prolonged exposure to mifepristone does not counteract the inhibitory effects of progestin therapy on endometrial proliferation. Estrogen and progesterone receptors may play an important role in these effects. PMID- 16311301 TI - NASA: back to eating seed corn. PMID- 16311302 TI - Avian influenza. China will attempt largest-ever animal vaccination campaign. PMID- 16311303 TI - U.S. budget. NIH set for tiny spending hike in 2006. PMID- 16311304 TI - Genetics. Expression of endorphin gene favored in human evolution. PMID- 16311305 TI - Medicine. Cancer-suppressing enzyme adds a link to type 2 diabetes. PMID- 16311306 TI - Society for Neuroscience meeting. Bats have a feel for flight. PMID- 16311307 TI - Society for Neuroscience meeting. Neuroscience society plans to leave New Orleans high and dry. PMID- 16311308 TI - Society for Neuroscience meeting. Computer game sharpens aging minds. PMID- 16311309 TI - Canada. New funding schemes aim to retain top academic talent. PMID- 16311310 TI - Higher education. U.S. plans suit to stop minority-only programs. PMID- 16311311 TI - After Katrina. Louisiana's wetlands struggle for survival. PMID- 16311312 TI - After Katrina. Tapping a river to restore and build up wetlands. PMID- 16311314 TI - Condensed-matter physics. Researchers turn up the heat in superconductivity hunt. PMID- 16311313 TI - After Katrina. New Orleans labs start their uncertain comeback. PMID- 16311315 TI - Space science. The question on the table: will Europe go to Mars? PMID- 16311316 TI - Making a rebuilt New Orleans sustainable. PMID- 16311317 TI - Problems of studying extinction risks. PMID- 16311318 TI - Benefits of a regional climate model. PMID- 16311319 TI - Proposed changes to biomedical funding. PMID- 16311320 TI - The paradox of radiation's effects. PMID- 16311321 TI - Public health. Violence against women. PMID- 16311322 TI - Cell signaling. Frizzled at the cutting edge of the synapse. PMID- 16311323 TI - Atmospheric science. Tiny bubbles tell all. PMID- 16311324 TI - Planetary science. Saturn's strangest ring becomes curiouser and curiouser. PMID- 16311325 TI - Developmental biology. Encountering microRNAs in cell fate signaling. PMID- 16311326 TI - The Phanerozoic record of global sea-level change. AB - We review Phanerozoic sea-level changes [543 million years ago (Ma) to the present] on various time scales and present a new sea-level record for the past 100 million years (My). Long-term sea level peaked at 100 +/- 50 meters during the Cretaceous, implying that ocean-crust production rates were much lower than previously inferred. Sea level mirrors oxygen isotope variations, reflecting ice volume change on the 10(4)- to 10(6)-year scale, but a link between oxygen isotope and sea level on the 10(7)-year scale must be due to temperature changes that we attribute to tectonically controlled carbon dioxide variations. Sea-level change has influenced phytoplankton evolution, ocean chemistry, and the loci of carbonate, organic carbon, and siliciclastic sediment burial. Over the past 100 My, sea-level changes reflect global climate evolution from a time of ephemeral Antarctic ice sheets (100 to 33 Ma), through a time of large ice sheets primarily in Antarctica (33 to 2.5 Ma), to a world with large Antarctic and large, variable Northern Hemisphere ice sheets (2.5 Ma to the present). PMID- 16311327 TI - Singing icebergs. AB - Sustained harmonic tremor signals were recorded by the seismographs of the German Neumayer Base seismological network in western Dronning Maud Land, Antarctica. These tremor episodes, lasting up to 16 hours, were recorded up to 820 kilometers from the source. Their spectra show narrow peaks with fundamental frequencies ranging from 0.5 to 6 hertz, more than 30 integer harmonic overtones, and frequency gliding, resembling volcanic tremor. Frequency-wave number analysis suggested a moving source, which was recognized as iceberg B-09A traveling along the coast of eastern Antarctica. The most probable tremor sources are fluid-flow induced vibrations inside the iceberg's tunnel/crevasse systems. PMID- 16311328 TI - Cassini discovers a kinematic spiral ring around Saturn. AB - Since the time of the Voyager flybys of Saturn in 1980-1981, Saturn's eccentric F ring has been known to be accompanied on either side by faint strands of material. New Cassini observations show that these strands, initially interpreted as concentric ring segments, are in fact connected and form a single one-arm trailing spiral winding at least three times around Saturn. The spiral rotates around Saturn with the orbital motion of its constituent particles. This structure is likely the result of differential orbital motion stretching an initial cloud of particles scattered from the dense core of the F ring. Different scenarios of formation, implying ringlet-satellite interactions, are explored. A recently discovered moon candidate, S/2004 S6, is on an orbit that crosses the F ring core at the intersection of the spiral with the ring, which suggests a dynamical connection between S/2004 S6 and the spiral. PMID- 16311329 TI - Encoding electronic properties by synthesis of axial modulation-doped silicon nanowires. AB - We describe the successful synthesis of modulation-doped silicon nanowires by achieving pure axial elongation without radial overcoating during the growth process. Scanning gate microscopy shows that the key properties of the modulated structures-including the number, size, and period of the differentially doped regions-are defined in a controllable manner during synthesis, and moreover, that feature sizes to less than 50 nanometers are possible. Electronic devices fabricated with designed modulation-doped nanowire structures demonstrate their potential for lithography-independent address decoders and tunable, coupled quantum dots in which changes in electronic properties are encoded by synthesis rather than created by conventional lithography-based techniques. PMID- 16311330 TI - Super-compressible foamlike carbon nanotube films. AB - We report that freestanding films of vertically aligned carbon nanotubes exhibit super-compressible foamlike behavior. Under compression, the nanotubes collectively form zigzag buckles that can fully unfold to their original length upon load release. Compared with conventional low-density flexible foams, the nanotube films show much higher compressive strength, recovery rate, and sag factor, and the open-cell nature of the nanotube arrays gives excellent breathability. The nanotube films present a class of open-cell foam structures, consisting of well-arranged one-dimensional units (nanotube struts). The lightweight, highly resilient nanotube films may be useful as compliant and energy-absorbing coatings. PMID- 16311331 TI - The nature of aqueous tunneling pathways between electron-transfer proteins. AB - Structured water molecules near redox cofactors were found recently to accelerate electron-transfer (ET) kinetics in several systems. Theoretical study of interprotein electron transfer across an aqueous interface reveals three distinctive electronic coupling mechanisms that we describe here: (i) a protein mediated regime when the two proteins are in van der Waals contact; (ii) a structured water-mediated regime featuring anomalously weak distance decay at relatively close protein-protein contact distances; and (iii) a bulk water mediated regime at large distances. Our analysis explains a range of otherwise puzzling biological ET kinetic data and provides a framework for including explicit water-mediated tunneling effects on ET kinetics. PMID- 16311332 TI - Stable carbon cycle-climate relationship during the Late Pleistocene. AB - A record of atmospheric carbon dioxide (CO2) concentrations measured on the EPICA (European Project for Ice Coring in Antarctica) Dome Concordia ice core extends the Vostok CO2 record back to 650,000 years before the present (yr B.P.). Before 430,000 yr B.P., partial pressure of atmospheric CO2 lies within the range of 260 and 180 parts per million by volume. This range is almost 30% smaller than that of the last four glacial cycles; however, the apparent sensitivity between deuterium and CO2 remains stable throughout the six glacial cycles, suggesting that the relationship between CO2 and Antarctic climate remained rather constant over this interval. PMID- 16311333 TI - Atmospheric methane and nitrous oxide of the Late Pleistocene from Antarctic ice cores. AB - The European Project for Ice Coring in Antarctica Dome C ice core enables us to extend existing records of atmospheric methane (CH4) and nitrous oxide (N2O) back to 650,000 years before the present. A combined record of CH4 measured along the Dome C and the Vostok ice cores demonstrates, within the resolution of our measurements, that preindustrial concentrations over Antarctica have not exceeded 773 +/- 15 ppbv (parts per billion by volume) during the past 650,000 years. Before 420,000 years ago, when interglacials were cooler, maximum CH4 concentrations were only about 600 ppbv, similar to lower Holocene values. In contrast, the N2O record shows maximum concentrations of 278 +/- 7 ppbv, slightly higher than early Holocene values. PMID- 16311334 TI - Assistance of microbial glycolipid antigen processing by CD1e. AB - Complexes between CD1 molecules and self or microbial glycolipids represent important immunogenic ligands for specific subsets of T cells. However, the function of one of the CD1 family members, CD1e, has yet to be determined. Here, we show that the mycobacterial antigens hexamannosylated phosphatidyl-myo inositols (PIM6) stimulate CD1b-restricted T cells only after partial digestion of the oligomannose moiety by lysosomal alpha-mannosidase and that soluble CD1e is required for this processing. Furthermore, recombinant CD1e was able to bind glycolipids and assist in the digestion of PIM6. We propose that, through this form of glycolipid editing, CD1e helps expand the repertoire of glycolipidic T cell antigens to optimize antimicrobial immune responses. PMID- 16311335 TI - Vertebrate-type intron-rich genes in the marine annelid Platynereis dumerilii. AB - Previous genome comparisons have suggested that one important trend in vertebrate evolution has been a sharp rise in intron abundance. By using genomic data and expressed sequence tags from the marine annelid Platynereis dumerilii, we provide direct evidence that about two-thirds of human introns predate the bilaterian radiation but were lost from insect and nematode genomes to a large extent. A comparison of coding exon sequences confirms the ancestral nature of Platynereis and human genes. Thus, the urbilaterian ancestor had complex, intron-rich genes that have been retained in Platynereis and human. PMID- 16311336 TI - SMEDWI-2 is a PIWI-like protein that regulates planarian stem cells. AB - We have identified two genes, smedwi-1 and smedwi-2, expressed in the dividing adult stem cells (neoblasts) of the planarian Schmidtea mediterranea. Both genes encode proteins that belong to the Argonaute/PIWI protein family and that share highest homology with those proteins defined by Drosophila PIWI. RNA interference (RNAi) of smedwi-2 blocks regeneration, even though neoblasts are present, irradiation-sensitive, and capable of proliferating in response to wounding; smedwi-2(RNAi) neoblast progeny migrate to sites of cell turnover but, unlike normal cells, fail at replacing aged tissue. We suggest that SMEDWI-2 functions within dividing neoblasts to support the generation of cells that promote regeneration and homeostasis. PMID- 16311337 TI - Representation of action-specific reward values in the striatum. AB - The estimation of the reward an action will yield is critical in decision-making. To elucidate the role of the basal ganglia in this process, we recorded striatal neurons of monkeys who chose between left and right handle turns, based on the estimated reward probabilities of the actions. During a delay period before the choices, the activity of more than one-third of striatal projection neurons was selective to the values of one of the two actions. Fewer neurons were tuned to relative values or action choice. These results suggest representation of action values in the striatum, which can guide action selection in the basal ganglia circuit. PMID- 16311338 TI - Nucleus accumbens long-term depression and the expression of behavioral sensitization. AB - Drug-dependent neural plasticity related to drug addiction and schizophrenia can be modeled in animals as behavioral sensitization, which is induced by repeated noncontingent or self-administration of many drugs of abuse. Molecular mechanisms that are critical for behavioral sensitization have yet to be specified. Long term depression (LTD) of alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptor (AMPAR)-mediated synaptic transmission in the brain has been proposed as a cellular substrate for learning and memory. The expression of LTD in the nucleus accumbens (NAc) required clathrin-dependent endocytosis of postsynaptic AMPARs. NAc LTD was blocked by a dynamin-derived peptide that inhibited clathrin-mediated endocytosis or by a GluR2-derived peptide that blocked regulated AMPAR endocytosis. Systemic or intra-NAc infusion of the membrane-permeable GluR2 peptide prevented the expression of amphetamine-induced behavioral sensitization in the rat. PMID- 16311339 TI - Wingless signaling at synapses is through cleavage and nuclear import of receptor DFrizzled2. AB - Wingless secretion provides pivotal signals during development by activating transcription of target genes. At Drosophila synapses, Wingless is secreted from presynaptic terminals and is required for synaptic growth and differentiation. Wingless binds the seven-pass transmembrane DFrizzled2 receptor, but the ensuing events at synapses are not known. We show that DFrizzled2 is endocytosed from the postsynaptic membrane and transported to the nucleus. The C terminus of DFrizzled2 is cleaved and translocated into the nucleus; the N-terminal region remains just outside the nucleus. Translocation of DFrizzled2-C into the nucleus, but not its cleavage and transport, depends on Wingless signaling. We conclude that, at synapses, Wingless signal transduction occurs through the nuclear localization of DFrizzled2-C for potential transcriptional regulation of synapse development. PMID- 16311340 TI - My approach to superficial inflammatory dermatoses. AB - Superficial inflammatory dermatoses are very common and comprise a wide, complex variety of clinical conditions. Accurate histological diagnosis, although it can sometimes be difficult to establish, is essential for clinical management. Knowledge of the microanatomy of the skin is important to recognise the variable histological patterns of inflammatory skin diseases. This article reviews the non vesiculobullous/pustular inflammatory superficial dermatoses based on the compartmental microanatomy of the skin. PMID- 16311341 TI - Involvement of MMP-7 in invasion of pancreatic cancer cells through activation of the EGFR mediated MEK-ERK signal transduction pathway. AB - AIMS: To clarify the involvement of matrix metalloproteinase-7 (MMP-7) in cell dissociation and the subsequent invasion of pancreatic cancer cells. METHODS: Western blotting, in vitro invasion assay, immunocytochemistry, and immunohistochemistry were performed in pancreatic cancer cell lines or pancreatic cancer tissue. RESULTS: The active form of the MMP-7 protein was expressed exclusively in the conditioned medium of dissociated (PC-1.0 and AsPC-1) pancreatic cancer cells, whereas proMMP-7 protein was only detected in the conditioned medium of non-dissociated (PC-1 and Capan-2) cells. Both intracellular and conditioned medium localised MMP-7 was greatly reduced by treatment with the epidermal growth factor receptor (EGFR) inhibitor AG1478 and the mitogen activated protein kinase kinase (MEK) inhibitor U0126 in pancreatic cancer cells. MMP-7 treatment significantly induced the disruption of tight junction (TJ) structures and subsequent cell dissociation, and activation of the EGFR mediated MEK- ERK (extracellular signal regulated protein kinase) signalling pathway in the non-dissociated pancreatic cancer cells. Moreover, the strong in vitro invasiveness of dissociated cells was inhibited by AG1478 and U0126 treatment, whereas the weak invasiveness of non-dissociated cells was apparently induced by MMP-7 treatment. In addition, MMP-7 expression was stronger at the invasive front than at the centre of human pancreatic tumours. CONCLUSION: MMP-7 is involved in cell dissociation and the subsequent invasion of pancreatic cancer cells. It induces the disruption of TJ structures and forms a positive feedback loop with activation of the EGFR mediated MEK-ERK signalling pathway. PMID- 16311342 TI - FAT, E-cadherin, beta catenin, HER 2/neu, Ki67 immuno-expression, and histological grade in intrahepatic cholangiocarcinoma. AB - AIM: To identify surrogate prognostic markers in intrahepatic cholangiocarcinoma (IHCC). METHODS: Thirty one cases of IHCC were graded and immunostained for FAT, Ki67, E-cadherin, beta catenin, and HER 2/neu. RESULTS: Twenty two cases were high grade and 27 had high Ki67 counts. Strong membranous staining of HER 2/neu was found in 10 tumours and reduced membranous E-cadherin and beta catenin in 19 and 18 tumours, respectively. Nuclear localisation of beta catenin was identified in five tumours and 22 showed weak cytoplasmic staining of FAT. Strong HER 2/neu and weak FAT immuno-expression were significantly correlated with high histological grade (p=0.01) and high Ki67 index (p=0.03). Upregulation of HER 2/neu was also significantly associated with nuclear localisation of beta catenin (p=0.01). Reduced membranous beta catenin was significantly related to reduced membranous E-cadherin (p=0.03), weak staining for FAT (p=0.01), and nuclear translocation of beta catenin (p=0.04). CONCLUSIONS: Reduced immuno-expression of E-cadherin and FAT at their normal membranous location may be potential prognostic markers, and the overexpression of HER 2/neu and beta catenin nuclear translocation may have a role in cholangiocarcinogenesis. PMID- 16311343 TI - mRNA expression of genes involved in lipid efflux and matrix degradation in occlusive and ectatic atherosclerotic disease. AB - BACKGROUND: Atherosclerotic plaque behaviour is influenced by intra-plaque inflammation, matrix turnover, and the lipid core volume. Peroxisome proliferator activated receptor gamma (PPARgamma) modulates atherosclerosis by its anti inflammatory and anti-protease activity. PPARgamma promotes lipid efflux through the liver X receptor alpha (LXRalpha) and the ATP binding cassette transporter A1 (ABCA1). Matrix metalloproteinase 9 (MMP-9) and cyclooxygenase 2 (COX-2) are implicated in plaque instability. AIMS: To assess the expression of these genes in occlusive and ectatic atherosclerotic disease to determine the relation between genes involved in lipid efflux and matrix degradation. METHODS: Carotid endarterectomy specimens from 16 patients and aneurysm tissue from 16 patients undergoing abdominal aortic aneurysm repair were used. Inferior mesenteric arteries from colectomy specimens from 12 patients served as controls. Total RNA was extracted from pulverised tissue and reverse transcribed into cDNA. Quantitative real time polymerase chain reaction (PCR) was performed using fluorescently labelled probes for ABCA1, LXRalpha, PPARgamma, COX-2, and MMP-9. RESULTS: PPARgamma expression was significantly lower in both occlusive and ecstatic atherosclerotic disease (p<0.001), whereas LXRalpha and ABCA1 expression was significantly increased (p<0.01). MMP-9 expression was significantly increased in diseased tissues (p<0.0001), and values were highest in occlusive disease (p<0.01). The increases in ABCA1 and MMP-9 mRNA were significantly correlated in diseased tissues (p<0.01, r=0.71 and r=0.78). COX-2 expression was increased in ectatic but low in occlusive disease (p<0.01). CONCLUSION: This observational study suggests a role for therapeutic upregulation of PPARgamma, which could potentially upregulate lipid efflux through ABCA1 and inhibit matrix degradation through inhibition of MMP-9. PMID- 16311344 TI - Histological examination has a major impact on macroscopic necropsy diagnoses. AB - BACKGROUND: Necropsy is the gold standard for clinicopathological discrepancy studies and epidemiological surveys. Inadequate sampling or lack of tissue may hamper the final interpretation and quality of the necropsy. AIM: To compare the histological and gross necropsy diagnoses of different organs. METHODS: A retrospective comparison of the provisional reports (gross findings only) and the final reports (after histological examination) of the necropsies performed at the department of pathology of the Hospital das Clinicas, Sao Paulo University, Brazil, a large tertiary care complex, in 2001. The total number of diagnoses listed for the lungs, heart, liver, pancreas, kidneys, and spleen were calculated. Findings were categorised into concordant/refined diagnosis, discordant/additional diagnosis, histology needed, and inconclusive. RESULTS: Three hundred and seventy one postmortem reports were analysed. There were 214 men and 157 women, with a mean age of 50.3 years, ranging from 1 to 92. The lung received the highest number (954) and the pancreas the lowest number (390) of diagnoses. The highest frequencies of discrepancies between the gross and microscopic findings were found in the lung and the liver: 38.7% and 35.1%, respectively. The brain had the lowest frequency of discrepancies. In a small number of cases, the final diagnosis could only be achieved through microscopic analysis, with the highest frequency being found in the kidneys (8.5%). CONCLUSIONS: Histological analysis has a major impact on previously performed gross diagnosis at necropsy, especially in the lungs, liver, and kidneys. Adequate sampling and histological analysis are important for necropsy quality. PMID- 16311345 TI - Characterisation of adherens and tight junctional molecules in normal animal larynx; determining a suitable model for studying molecular abnormalities in human laryngopharyngeal reflux. AB - BACKGROUND: The disruption of intercellular junctions in the larynx is a pathological feature of laryngopharyngeal reflux (LPR). Good experimental models are necessary to gain greater insight into the molecular mechanisms and alterations that result from abnormal exposure of the laryngeal epithelium to acid refluxate. AIMS: To characterise laryngeal tissues from different species to determine the most suitable for use in experimental studies of LPR. METHODS: Human and non-human laryngeal tissues (mouse, rat, guinea pig, porcine, and rabbit) were studied. Histological characterisation was performed by light microscopy. The expression and subcellular localisation of adherens junctional molecules (E-cadherin and beta catenin) was evaluated by immunohistochemistry, and tight junction molecules (occludin and zonula occludens 1 (ZO-1)) by western blotting. The ultrastructural features of porcine and human tissue were assessed by electron microscopy. RESULTS: Porcine tissue revealed both respiratory-type and stratified squamous epithelium, as seen in the human larynx. The expression and subcellular localisation of the E-cadherin-catenin complex was detected in all species except mouse and rat. The pattern of ZO-1 and occludin expression was preserved in all species. CONCLUSION: The expression of intercellular junctional complexes in porcine epithelium is similar to that seen in humans. These results confirm the suitability of these species to study molecular mechanisms of LPR in an experimental system. PMID- 16311346 TI - Extensive intestinal metaplasia in gastric carcinoma and in other lesions requiring surgery: a study of 3,421 gastrectomy specimens from dwellers of the Atlantic and Pacific basins. AB - BACKGROUND: Extensive intestinal metaplasia (EIM) has been reported in gastrectomies from patients dwelling in the Pacific and Atlantic basins. AIMS: To compare all the results in an attempt to explain the findings. METHOD: All sections from 3,421 gastrectomies were reviewed at various hospitals: 1946 in the Atlantic and 1475 in the Pacific basin. Sections with EIM showed IM encompassing one or more entire low power field (>or=5 mm in length/section) in one or more section. RESULTS: In the Atlantic basin, EIM was present in 18.8% (153 of 814) of specimens with intestinal carcinoma (IC) and in 10.3% (65 of 630) of those with diffuse carcinoma (DC). In the Pacific basin, EIM was found in 62.9% (412 of 655) of gastrectomies with IC and in 33.3% (160 of 481) of those with DC. The numbers of specimens with EIM were significantly higher in the Pacific than in the Atlantic basin for both carcinoma phenotypes, particularly among elderly patients (>or=60 years). CONCLUSIONS: The proportion of gastrectomies with EIM was higher among populations at a higher gastric cancer risk than in those with a lower cancer risk. EIM was mostly associated with IC rather than DC or with miscellaneous gastric diseases (841 control gastrectomies) in both basins. The proportion of gastrectomies with EIM was significantly higher in Vancouver than in New York and in Santiago de Chile than in Buenos Aires, even though these populations reside at approximately the same geographical latitude, but in different basins. Environmental factors seem to accelerate the evolution of EIM. PMID- 16311347 TI - Expression of CD1d in human scalp skin and hair follicles: hair cycle related alterations. AB - BACKGROUND: CD1d belongs to a family of antigen presenting molecules that are structurally and distantly related to the classic major histocompatibility complex class I (MHC I) proteins. However, unlike MHC I molecules, which bind protein antigens, CD1d binds to lipid and glycolipid antigens. CD1d is expressed by cells of lymphoid and myeloid origin, and by cells outside of the lymphoid and myeloid lineages, such as human keratinocytes of psoriatic skin. AIMS: To investigate whether CD1d is also expressed in sun exposed skin and in the immuno privileged anagen hair follicle. MATERIALS/METHODS: CD1d immunoreactivity was studied in human scalp skin and hair follicles of healthy women in situ by immunofluorescent and light microscopic immunohistology. Skin biopsies were obtained from normal human scalp containing mainly anagen VI hair follicles from women (age, 53-57 years) undergoing elective plastic surgery. RESULTS: CD1d showed strong immunostaining in human scalp skin epidermis, pilosebaceous units, and eccrine glands. In the epidermis, CD1d was strongly expressed by basal and granular keratinocytes. In hair follicles, CD1d was expressed in the epithelial compartment and showed hair cycle related alterations, with an increase in the anagen and a reduction in the catagen and telogen phases. CONCLUSIONS: These results suggest that CD1d plays a role in human scalp skin immunology and protection against lipid antigen rich infectious microbes. They also raise the question of whether keratinocytes of the immuno-privileged anagen hair follicle can present lipid antigens to natural killer T cells. These data could help provide new strategies for the manipulation of hair related disorders. PMID- 16311348 TI - Survey of clinical allergy services provided by clinical immunologists in the UK. AB - BACKGROUND: The UK National Health Service is failing to meet the need for diagnosis and treatment of allergic disorders, which are common and increasing in prevalence. The House of Commons select committee report on allergy services highlighted the inequalities and urgent need for investment. AIM: To survey the allergy workload provided by clinical immunologists to inform service planning and resource allocation. METHODS: The allergy services performed by clinical immunologists during a 12 month period from 1 April 2003 to 31 March 2004 were surveyed by means of a questionnaire via supra-regional audit groups. RESULTS: The immunology centres surveyed serve 32 million people and offer almost the complete repertoire of a specialised allergy service. There were large variations in clinic capacity, new referrals, appointment duration, and service configuration. Services were largely consultant delivered, but availability of joint clinics with paediatricians and anaesthetists was locally variable. Novel service delivery models utilising nurses and clinical assistants have been developed and merit further investigation. CONCLUSION: Consultant immunologists and trainees currently make a major contribution to the development and provision of specialised allergy services. Consultant immunologists will probably remain key providers of tertiary level allergy care in the UK in the long term (in line with other countries) and will be pivotal in supporting and developing the provision of equitable national access to specialist allergy services in a timely manner. Rapid progress in developing the new specialty of allergy and securing better access to services for patients in the short term will be best served by strengthening the collaborative relationship between allergists and clinical immunologists. PMID- 16311349 TI - Necrobiotic palisading granulomas associated with breast carcinoma. AB - AIMS: The presence of granulomas within the stroma of carcinomas and in the lymph nodes draining carcinomas has been well described. To date, however, there have been few studies examining the occurrence and relevance of necrobiotic granulomas occurring in association with breast carcinoma. METHODS/RESULTS: Four cases of breast carcinoma with necrobiotic granulomas were examined using periodic acid Schiff and Ziehl Neelsen stains for fungi and tubercle bacilli and with immunohistochemistry using CAM 5.2, cytokeratin 7, and cytokeratin AE1/3 for tumour cells. In one case the stroma was involved, in the other three cases the lymph nodes contained necrobiotic granulomas. In two of the cases, one with stromal and one with lymph node involvement, the necrobiotic granulomas contained necrotic tumour cells. CONCLUSION: In this study the features of four cases of breast carcinomas with necrobiotic granulomas are examined and their relevance explored. Close scrutiny of such granulomas is necessary to avoid underdiagnosis of metastatic disease. PMID- 16311350 TI - Light microscopic examination of scalp hair samples as an aid in the diagnosis of paediatric disorders: retrospective review of more than 300 cases from a single centre. AB - BACKGROUND: Microscopic examination of scalp hair can provide important diagnostic information in a range of paediatric conditions. It is a non-invasive and cost effective investigation, which is not widely performed. AIMS: To examine retrospectively the value of hair examination by light microscopy, including polarising microscopy, in a specialist paediatric pathology department during a 15 year period (1989-2004) and to describe the morphological abnormalities indicative of specific paediatric conditions. METHODS: Three hundred and twenty two hair samples were submitted. Microscopic changes were analysed in the light of clinical information categorised as: (1) erythroderma, (2) neurological impairment, (3) immunological/haematological defect, (4) ectodermal dysplasia, (5) abnormal hair only, and (6) non-specific/absent clinical details. RESULTS: Abnormalities were evident in 49% of the samples. In 25%, the changes were compatible with specific diagnoses including Menkes disease, Netherton's syndrome, trichothiodystrophy, Griscelli and Chediak-Higashi syndromes, monilethrix, uncombable hair, and loose anagen syndromes. In respect of the clinical presentation groups noted above, diagnostic changes were seen in 41%, 32%, 33%, 0%, 29%, and 0%, respectively. CONCLUSIONS: Morphological light microscopic examination of scalp hair is an inexpensive, rapid, and non-invasive investigation, which can provide valuable diagnostic information in a range of paediatric conditions. PMID- 16311351 TI - Frequent overexpression of epidermal growth factor receptor (EGFR) in mammary high grade ductal carcinomas with myoepithelial differentiation. AB - AIMS: To evaluate the expression of common biological markers and the epidermal growth factor receptor (EGFR) in mammary high grade ductal carcinomas with myoepithelial differentiation (DCMDs). MATERIALS/METHODS: Thirty DCMDs were clinicopathologically and immunohistochemically analysed and compared with 36 control cases of high grade conventional invasive ductal carcinoma (IDC). RESULTS: EGFR, HER2/neu, oestrogen receptor, progesterone receptor, and p53 expression was seen in 21, one, three, four, and 20 of the 30 DCMDs, compared with eight, nine, 18, 17, and five of the 36 conventional IDCs (p<0.05), respectively. In 16 of the 30 DCMDs, metastases were found in the brain, lung, bone, and liver, within a maximum of 47 months (mean, 13.9) after initial surgery, whereas only four of the 36 conventional IDCs metastasised to the lung and bone within a maximum of 27 months (mean, 18.0) after initial surgery (p=0.0001). There was a significant difference in disease free survival between DCMD and conventional IDC (p=0.001). EGFR was frequently overexpressed in DCMD compared with conventional IDC, whereas the expression of HER2/neu and hormone receptors was lower in DCMD. Fluorescent in situ hybridisation revealed that the mean EGFR to chromosome 7 centromere (CEP7) ratio of the 24 DCMD cases available for evaluation was 1.03, and EGFR gene amplification was not detected in the 21 DCMD cases with EGFR overexpression. CONCLUSION: Immunohistochemistry for myoepithelial markers and EGFR is useful for the accurate diagnosis and molecular target treatment of high grade DCMD. PMID- 16311352 TI - Blood film examination for vacuolated lymphocytes in the diagnosis of metabolic disorders; retrospective experience of more than 2,500 cases from a single centre. AB - BACKGROUND: A range of metabolic diseases can result in abnormal accumulation of metabolic byproducts, resulting in abnormal lymphocyte cytoplasmic vacuolation, identifiable on routine blood film examination. AIMS: This study retrospectively examines the usefulness of blood film examination for vacuolated lymphocytes in a specialist paediatric pathology department in relation to patient's age and presentation. It also describes specific diagnostic features in relation to specific classes of metabolic disease. METHODS: Retrospective review of a histopathology database to identify all blood films examined for the detection of vacuolated lymphocytes during a 15 year period (1989-2004). RESULTS: In total, 2,550 blood films were investigated. The median age at submission was 2 years (range, birth to 88), and>90% of samples were from children<18 years. The most common indications were developmental delay/regression, ataxia, seizures, and cardiomyopathy. Vacuolated lymphocytes were identified in 156 films (6.1%). The frequency of vacuolated lymphocytes varied with clinical presentation, with ophthalmic indications having the highest positive rate (40%). In cases with vacuolated lymphocytes, a wide range of underlying metabolic diagnoses was apparent, the most common being juvenile neuronal ceroid lipofuscinosis and acid maltase deficiency, which accounted for more than half of the diagnoses. CONCLUSIONS: The examination of blood films for lymphocyte vacuolation is clinically useful in patients with a history suggestive of metabolic disease. The test is cheap, rapid, minimally invasive, and provides first line screening, with some findings indicating clues to a specific underlying diagnosis. PMID- 16311353 TI - Maspin expression in carcinoma ex pleomorphic adenoma. AB - AIMS: To investigate the presence and distribution of the protein maspin in carcinoma ex pleomorphic adenoma (CXPA). METHODS: Maspin expression was studied by means of immunohistochemistry in 16 cases of CXPA, using the labelled polymer method. RESULTS: According to the extent of invasion, the tumours were subdivided into: intracapsular (five cases), minimally invasive (four cases), and invasive (seven cases). Twelve patients had carcinoma with only epithelial differentiation, whereas four had a malignant myoepithelial component. Non luminal cells in the duct-like structures of the remnant pleomorphic adenoma were strongly positive for maspin, whereas only a few luminal cells were immunopositive. A few positive cells were seen in the frequent hypocellular and hyalinised areas. Maspin was abundantly expressed, mainly in non-luminal cells, in transitional areas of CXPA with only epithelial differentiation. In frankly carcinomatous areas there was a gradual decrease in maspin expression. Almost all cells were maspin positive in CXPA with a myoepithelial component. When present, luminal cells were in general negative for maspin. CONCLUSIONS: When only epithelial cells undergo malignant transformation, maspin expression is gradually lost. In cases with a myoepithelial component, maspin expression is high, and this might be related to the tumour suppressor activity attributed to this cell. PMID- 16311354 TI - Comparison of the novel quantitative ARMS assay and an enriched PCR-ASO assay for K-ras mutations with conventional cytology on endobiliary brush cytology from 312 consecutive extrahepatic biliary stenoses. AB - BACKGROUND: Extrahepatic biliary stenosis (EBS) has malignant and benign causes. Patients with EBS are at risk of having or developing malignancy. Accurate diagnostic tests for early detection and surveillance are needed. The sensitivity of biliary cytology for malignancy is low. K-ras mutation analysis on brush cytology is a valuable adjunct, but specificity is low. A quantitative test for K ras mutations has been developed: the amplification refractory mutation system (ARMS). AIM: To assess the test characteristics and additional value of ARMS in diagnosing the cause of EBS. METHODS: Brush samples from endoscopic retrograde cholangiopancreatography were collected from 312 patients with EBS. K-ras mutation analysis was performed using ARMS-allele specific amplification was coupled with real time fluorescent detection of PCR products. Results were compared with conventional cytology and K-ras mutation analysis using allele specific oligonucleotide (ASO) hybridisation, and evaluated in view of the final diagnosis. RESULTS: The test characteristics of ARMS and ASO largely agreed. Sensitivity for detecting malignancy was 49% and 42%, specificity 93% and 88%, and positive predictive value (PPV) 96% and 91%, respectively. The sensitivity of ARMS and cytology combined was 71%, and PPV was 93%. The specificity of ARMS could be increased to 100% by setting limits for the false positives, but reduced sensitivity from 49% to 43%. CONCLUSIONS: ARMS can be considered supplementary to conventional cytology, and comparable to ASO in diagnosing malignant EBS. A specificity of 100% can be achieved with ARMS, which should be considered in the surveillance of patients at risk for pancreatic cancer. PMID- 16311355 TI - Unusual duodenal presentation of leishmaniasis. AB - This case report describes an atypical case of duodenal leishmaniasis in an elderly patient not infected with human immunodeficiency virus. Investigation of this 84 year old woman with a constitutional syndrome and dysphagia revealed anaemia of chronic disorder, a high erythrocyte sedimentation rate, and polyclonal hypergammaglobulinaemia. Abdominal ultrasonography revealed thickening of the stomach wall, which was seen to be inflamed during gastroscopy. Duodenal histology revealed numerous leishmania amastigotes within macrophages. This was confirmed by bone marrow biopsy and leishmania serology. This case report stresses the importance of atypical symptoms and the unusual location of visceral leishmaniasis, not only in immunodepressed patients, but also in elderly immunocompetent patients. PMID- 16311356 TI - Two cases of inflammatory muscle disease presenting with raised serum concentrations of troponin T. AB - Troponins T and I are highly sensitive markers of myocardial injury. However, non cardiac disorders, such as pulmonary embolism, renal failure, subarachnoid haemorrhage, sepsis, eclampsia, chemotherapy, and inflammatory muscle conditions (dermatomyositis and polymyositis), can also result in raised serum troponin concentrations. This article describes two cases that occurred within a month of each other in Craigavon Area Hospital, whereby conditions unrelated to myocardial ischaemia resulted in raised concentrations of cardiac markers. The first patient, in retrospect, underwent unnecessary investigation as an inpatient in the cardiac ward. Experience gained from this case led to more appropriate consultation and management of the second patient. PMID- 16311357 TI - A "crackleware" oesophagus. AB - This case report describes a 70 year old woman with excessive diffuse keratinisation of the oral cavity and oesophagus harbouring a squamous cell carcinoma. This excessive diffuse keratinisation of normally non-keratinised squamous epithelium could not be identified in normally non-keratinised epithelia in other parts of the body (the vagina), arguing against a genetic basis for this disorder. The term "crackleware" oesophagus was used to describe this entity, which has not been described previously in the English literature. PMID- 16311358 TI - Small cell sweat gland carcinoma of childhood. AB - Small cell sweat gland carcinoma appears to represent a very unusual histological type of sweat gland anlage tumour presenting in children. The differential diagnosis from other small blue cell tumours involving the skin is often difficult. The present report confirms the original observation describing two patients of 2 and 5 years of age harbouring cutaneous tumours. The histology of these lesions showed a monomorphic proliferation of small cells with a high mitotic rate and areas of necrosis. Immunohistochemically, the cells were negative for desmin, cytokeratin 7, cytokeratin 20, Cam 5.2, CD99, chromogranin, CD56, synaptophysin, and S-100, and focally positive for the pancytokeratin marker AE1/AE3, carcinoembryonic antigen (one case), and neurone specific enolase (one case). The prognosis of this type of tumour seems to be good. As more cases are added, the clinical pathological spectrum of the lesion will become better defined. PMID- 16311359 TI - Pure testicular carcinoid associated with intratubular germ cell neoplasia. AB - This case report describes for the first time a case of pure testicular carcinoid pre-aortic lymph node metastases in a 25 year old patient with carcinoid syndrome. The simultaneous occurrence of intratubular germ cell neoplasia in the surrounding testicular tissue was identified by OCT4 and placental-like alkaline phosphatase positivity. This confirmed that the tumour had a germ cell origin in the testis, rather than being a metastasis from an extragenital carcinoid. PMID- 16311360 TI - Sclerochoroidal calcification associated with Gitelman syndrome and calcium pyrophosphate dihydrate deposition. AB - Sclerochoroidal calcification is an uncommon condition. Metabolic evaluation and clinical examination are important to exclude associated systemic conditions such as the Bartter and Gitelman syndromes. It has been suggested that the lesions seen in sclerochoroidal calcification are calcium pyrophosphate dihydrate crystals. This report describes the first documented case in the UK of sclerochoroidal calcification associated with Gitelman syndrome and calcium pyrophosphate dihydrate deposition. PMID- 16311361 TI - Application of the FICTION technique for the simultaneous detection of immunophenotype and chromosomal abnormalities in routinely fixed, paraffin wax embedded bone marrow trephines. AB - The use of interphase fluorescence in situ hybridisation (FISH) to study cytogenetic abnormalities in routinely fixed paraffin wax embedded tissue has become commonplace over the past decade. However, very few studies have applied FISH to routinely fixed bone marrow trephines (BMTs). This may be because of the acid based decalcification methods that are commonly used during the processing of BMTs, which may adversely affect the suitability of the sample for FISH analysis. For the first time, this report describes the simultaneous application of FISH and immunofluorescent staining (the FICTION technique) to formalin fixed, EDTA decalcified and paraffin wax embedded BMTs. This technique allows the direct correlation of genetic abnormalities to immunophenotype, and therefore will be particularly useful for the identification of genetic abnormalities in specific tumour cells present in BMTs. The application of this to routine clinical practice will assist diagnosis and the detection of minimal residual disease. PMID- 16311362 TI - Postmortem diagnosis of chronic granulomatous disease: how worthwhile is it? AB - A previously healthy 11 year old boy died unexpectedly after a rapid course of progressive pneumonia. Postmortem microbiology and histopathology suggested an underlying diagnosis of chronic granulomatous disease. This was confirmed by neutrophil oxidative burst and gene mutation analysis of other family members, one of whom benefited from early bone marrow transplantation. PMID- 16311363 TI - Metastasis of a caecal neuroendocrine carcinoma to the thyroid gland. PMID- 16311364 TI - Poorly differentiated hepatocellular carcinoma with unusual tubular structures. PMID- 16311365 TI - Liesegang rings in inflammatory breast lesions. PMID- 16311366 TI - Expression of HIF-1 alpha in human tumours. PMID- 16311368 TI - Family Practice and the new era of electronic publishing. PMID- 16311367 TI - Congenital bronchogenic cyst in the gastric mucosa. PMID- 16311369 TI - Evaluation of daptomycin activity against Staphylococcus aureus in an in vitro pharmacodynamic model under normal and simulated impaired renal function. AB - OBJECTIVES: Daptomycin is a lipopeptide antimicrobial that is primarily excreted by the kidney. We examined daptomycin bactericidal activity in an in vitro pharmacodynamic model (IVPM) under normal and simulated impaired renal function against methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin resistant S. aureus (MRSA). METHODS: Two clinical strains MSSA-1199 and MRSA-494 were used in an IVPM. MICs and MBCs were determined according to CLSI. Daptomycin free concentrations were simulated that corresponded to dose regimens of 4, 6 and 8 mg/kg every 24 h at 8 h t(1/2) (Scheme I) and every 48 h at 30 h t(1/2) (Scheme II). In addition, we simulated daptomycin free concentrations corresponding to fractional dose regimens of 2, 3 and 4 mg/kg every 24 h at 30 h t(1/2) (Scheme III). The targeted C(max free)/MIC for 2, 3, 4, 6 and 8 mg/kg against MSSA-1199 ranged from 5.2 to 21.2. The targeted C(max free)/MIC for 2, 3, 4, 6 and 8 mg/kg against MRSA-494 ranged from 10.4 to 42.2. The targeted AUC(free)/MIC for Schemes I, II and III against MSSA-1199 ranged from 94 to 392. The targeted AUC(free)/MIC for Schemes I, II and III against MRSA-494 ranged from 188 to 581. Bactericidal activity and the potential for resistance were determined over 96 h. All models were completed in triplicate. RESULTS: Daptomycin MICs (MBCs) for MSSA-1199 and MRSA-494 were 0.5 (1.0) mg/L and 0.25 (0.25) mg/L, respectively. Daptomycin 6 and 8 mg/kg at both 8 and 30 h t(1/2) achieved 99.9% kill as early as 1 h. Daptomycin 4 mg/kg achieved 99.9% kill as early as 1 h when given at 8 and 30 h t(1/2) but was not maintained to an endpoint of 96 h (P > 0.05). CONCLUSIONS: Overall, there was no difference in kill noted for daptomycin regimens at 4, 6 and 8 mg/kg every 24 h at 8 h t(1/2) versus every 48 h at 30 h t(1/2). Fractional doses of daptomycin at 30 h t(1/2) were inferior to daptomycin regimens of 4, 6 and 8 mg/kg administered every 48 h (P = 0.03). PMID- 16311373 TI - DEFRA considers UK rabies control import policies. PMID- 16311374 TI - Tackling MRSA in animals and humans. PMID- 16311375 TI - Emerging zoonotic epidemics in the interconnected global community. AB - The rate at which epidemics of zoonotic disease in humans have surfaced over the past 25 years has shaken--some would say shattered--the assumption that zoonotic diseases are under control, says Paul Gibbs. In this review he analyses the global factors that have led to the increased emergence of zoonotic diseases, sketches several recent epidemics (and where relevant, their relationship to bioterrorism), discusses the lessons learned, and concludes by outlining an agenda for action. PMID- 16311376 TI - Integrated working. PMID- 16311377 TI - Crossing the boundaries. AB - In the past, research on zoonotic diseases has been fragmented. Teresa Belcher and Diane Newell discuss the formation of Med-Vet-Net, a "virtual institute", which aims to integrate activity between researchers in human and veterinary medicine across the European Union. PMID- 16311378 TI - Identifying hazards, assessing the risks. PMID- 16311379 TI - Veterinarians and food safety. AB - Since the early days of veterinary science, vets have played an important part in protecting public health through controls over the safety of food of animal origin. Traditionally, public health has been treated as a topic separate from the mainstream veterinary subjects of diagnosis and treatment of animal disease. However, there is now increasing awareness of the interconnection between animal and human health, and between the work of vets in production animal practice and those in food safety and other disciplines, as Kenneth Clarke explains. PMID- 16311380 TI - Therapeutic applications of the human-companion animal bond. PMID- 16311381 TI - Working in parallel, learning in parallel? PMID- 16311382 TI - Clinical audit--learning from the medical profession. PMID- 16311383 TI - Survey of zoonoses recorded in Scotland between 1993 and 2002. AB - All the human and animal laboratory reports of zoonoses sent to Health Protection Scotland between 1993 and 2002 were identified. There were 24,946 reports from veterinary laboratories, and 94,718 (20 per cent) of the 468,214 reports from medical laboratories were considered to be zoonotic. The most common reports of zoonoses from people were Campylobacter, Salmonella, Cryptosporidium and Giardia species and Escherichia coli o157. The most common reports of zoonoses from animals were Salmonella, Cryptosporidium, Chlamydia and Campylobacter species and Mycobacterium avium paratuberculosis. For all the zoonoses in people, the National Health Service Board areas Borders, Dumfries and Galloway, Forth Valley, Grampian, Lanarkshire and Lothian had a higher than expected standardised incidence rate of infection, whereas Ayrshire and Arran, Fife, Greater Glasgow, Shetland, Tayside and Western Isles had a lower than expected rate. The organisms and diseases considered to be new and emerging were Rhodococcus species, Cyclospora cayetanensis, Leishmania species, Pneumocystis carinii (jiroveci) and bovine spongiform encephalopathy/variant Creutzfeldt-Jakob disease. PMID- 16311384 TI - Prevalence, incidence and geographical distribution of serovars of Salmonella on dairy farms in England and Wales. AB - A study of randomly selected dairy farms in England and Wales was made between October 1999 and February 2001 to estimate the prevalence and incidence of Salmonella serovars. The farms were enrolled through five milk-buying companies, which represented 63 per cent of the dairy farms in England and Wales, and they were sampled on up to four occasions (449 farms at visit 1, 272 farms at visit 2, 251 farms at visit 3 and 243 farms at visit 4). In total, 19,296 samples of pooled faecal pats and slurry were collected. The farm-specific prevalence of all serovars of Salmonella ranged from 12.1 per cent (95 per cent confidence interval [CI] 8.2 to 16.0 per cent) to 24.7 per cent (95 per cent CI 19.4 to 30.1 per cent) at each visit. The most common serovars identified were Salmonella Dublin (3.7 to 6.6 per cent farm-specific prevalence at each visit), Salmonella Agama (1.8 to 7.6 per cent) and Salmonella Typhimurium (2.6 to 4.1 per cent) The prevalence varied by region and month of sampling and increased in late summer. The incidence rate of all serovars of Salmonella was 0.43 (95 per cent CI 0.34 to 0.54) cases per farm-year at risk. There was no significant difference between the incidence rates of the common serovars S Typhimurium (0.07), S Dublin (0.06) and S Agama (0.13). A total of 29 Salmonella serovars were isolated. Few of the isolates were resistant to the 16 antimicrobial agents tested, except the isolates of S Typhimurium dt104, of which 67.9 per cent were resistant to at least five of them. PMID- 16311385 TI - Serosurvey for West Nile virus in horses in southern France. PMID- 16311386 TI - Review of human rabies cases in the UK and in Germany. PMID- 16311390 TI - Prescribing under the new regulations. PMID- 16311391 TI - Riddles of the sphynx. PMID- 16311393 TI - CPR--the vanishing competency. PMID- 16311395 TI - Prevent unhealthy work environments. PMID- 16311396 TI - Advance directives in New Mexico. PMID- 16311397 TI - Intensive spiritual care: a case study. PMID- 16311398 TI - End-of-life decision making in intensive care. PMID- 16311399 TI - Decreasing parental stress in the pediatric intensive care unit: one unit's experience. PMID- 16311400 TI - Ask the experts. PMID- 16311401 TI - Is malpractice insurance important? PMID- 16311402 TI - Going through the wars: a passage in time. PMID- 16311403 TI - Functional evidence for a small and rigid active site in a high fidelity DNA polymerase: probing T7 DNA polymerase with variably sized base pairs. AB - Hypotheses on the origins of high fidelity in replicative DNA polymerases have recently focused on the importance of geometric or steric effects in this selectivity. Here we reported a systematic study of the effects of base pair size in T7 DNA polymerase (pol), the replicative enzyme for bacteriophage T7. We varied base pair size in very small (0.25 A) increments by use of a series of nonpolar thymidine shape mimics having gradually increasing size. Steady-state kinetics were evaluated for the 5A7A exonuclease-deficient mutant in a 1:1 complex with thioredoxin. For T7 pol, we studied insertion of natural nucleotides opposite variably sized T analogs in the template and, conversely, for variably sized dTTP analogs opposite natural template bases. The enzyme displayed extremely high selectivity for a specific base pair size, with drops in efficiency of as much as 280-fold for increases of 0.4 A beyond an optimum size approximating the size of a natural pair. The enzyme also strongly rejected pairs that were smaller than the optimum by as little as 0.3 A. The size preferences with T7 DNA pol were generally smaller, and the steric rejection was greater than DNA pol I Klenow fragment, correlating with the higher fidelity of the former. The hypothetical effects of varied active site size and rigidity are discussed. The data lend direct support to the concept that active site tightness is a chief determinant of high fidelity of replicative polymerases and that a less rigid (looser) and larger active site can lead to lower fidelity. PMID- 16311405 TI - Treatment of pituitary tumors: history. AB - Over the past century our understanding of the pituitary disease has undergone an amazing evolution, accompanied by impressive advances in the therapeutic approaches to pituitary tumors. This historical review aims to provide insights into the contributions of key medical and scientific pioneers, their occasionally serendipitous discoveries, and the lively global debates, which have ultimately improved the therapeutic targets and the long-term outcome of these patients. The development of the three main modalities is discussed (surgery, irradiation, pharmacotherapy). More recent experimental data, which may provide a path to a stronger therapeutic armamentarium for these undoubtedly challenging tumors, are also briefly reported. PMID- 16311406 TI - The history of the treatment of pituitary adenomas. AB - The immense history leading to our current understanding and treatment of pituitary pathology is inextricably linked to the evolution of the understanding of the numerous functions of the hypophysis cerebri as the "master gland" of the endocrine system. When the anatomists of old encountered this small organ sequestered "like a nugget in the innermost of Chinese boxes" at the base of the brain, they had no inkling of its importance in the control of multiple target organs in the human body. It would ultimately take two millennia and a vast amount of clinical and laboratory research for its role in the body to eventually become more completely appreciated. PMID- 16311407 TI - Classification and pathology of pituitary tumors. AB - Pituitary adenomas originating in adenohypophysial cells represent the most common neoplasms of the sella turcica. The pathologist's goal is the optimal diagnosis and classification of pituitary adenomas. Lack of clinicopathological correlations in the past classification of pituitary adenomas, which was based on the tinctorial properties of adenoma cells, limited the importance of histological diagnosis. Morphologic separation of pituitary cells by electron microscopy provided fundamental knowledge to classify pituitary adenomas. Immunohistochemistry represents the gold standard of the current classification. Combined morphologic and immunohistochemical diagnostic approaches resulted in the clinicopathologic classification of pituitary adenomas. The WHO classification of 2004 is based on morphologic features and takes into consideration findings from imaging procedures and clinical symptoms. Morphologic characterization of pituitary tumors and correlation of the hormone product with hormone secretion provides the clinician with useful information. In addition, the utility of tumor markers offers objective information in managing the patient and predicting responses to specific treatment. The Ki-67 labeling index (LI) is widely used for it correlates with invasiveness and probably prognosis. Adenomas showing increased ( >3%) LI and extensive p53 immunoreactivity should be termed "atypical adenomas" suggesting aggressive potential or malignant transformation. Morphologic separation of adenoma from carcinoma is not feasible. The term pituitary carcinoma should be exclusively applied when cerebrospinal and/or systemic metastases are definite. PMID- 16311408 TI - Recent progress in studies of pituitary tumor pathogenesis. AB - The mechanisms of tumorigenesis of the human pituitary have been elucidated to a limited extent. Classically, pituitary tumor formation was shown to be induced by thyroidectomy and estrogen administration. Molecular biological and immunohistochemical studies have revealed several aspects of pituitary tumorigenesis. Translineage cell differentiation has been shown to be induced by the aberrant expression of transcription factors and co-factors, such as Pit-1, Prop-1, and estrogen receptor. Defects or overexpression of cell cycle regulators, such as CDK inhibitors, PTTG, and GADD45gamma, result in the abnormal proliferation of pituitary cells. Recently, epigenetic regulation has been suggested to be related to pituitary tumor formation. This article presents a review and update of recent progress in studies of the development and differentiation of pituitary tumors. PMID- 16311409 TI - Genetics and proteomics of pituitary tumors. AB - Genetics and proteomics determine structure and function of normal tissues, and the molecular alterations that underlie tumorigenesis result in changes in these aspects of tissue biology in neoplasms. We review the known genetic alterations in pituitary tumors. These include the oncogenic Gsalpha protein (GSP)-activating mutations, and pituitary tumor-derived fibroblast growth factor receptor-4 (ptd FGFR4), as well as tumor suppressor gene mutations associated with multiple endocrine neoplasia type 1 (MEN1). Other candidates identified from expression profiling include pituitary tumor-transforming gene (PTTG), GADD45, and bone morphogenic protein (BMP)4. Proteomic changes in pituitary tumors include classical alterations identified by immunohistochemistry as well as epigenetic reductions in p27. The underlying mechanisms for dysregulated cell adhesive molecules including cadherins and FGFRs are reviewed. The combined use of genetic and proteomic approaches will enhance novel drug therapeutic development. PMID- 16311410 TI - DNA microarrays: recent developments and applications to the study of pituitary tissues. AB - Many new techniques are rapidly being developed and applied to the study of normal and neoplastic pituitary. DNA microarrays are a uniquely efficient method for simultaneously assessing the expression levels of thousands of genes, identifying disease subphenotypes, and predicting disease progression. This article reviews the utility of DNA microarray-based tumor profiling including recent developments and applications to pituitary biology in order to demonstrate how these new techniques are providing insights about basic aspects, clinical knowledge, and pharmacologic knowledge of the pituitary gland and about pituitary tumors. PMID- 16311411 TI - Pituitary tumors: prognostic indicators. AB - Many factors influence the proliferation of pituitary adenomas: angiogenesis, apoptosis, growth factors, oncogenes, tumor suppressor genes, and hormone receptors. These elements can be demonstrated by immunohistochemistry and/or molecular pathology but no single factor can be used for determination of biological behavior resp. prognosis. Pituitary adenomas can be enclosed or invasive and may be very large or may be microadenomas, but the most important point for prognosis is the total resection in the first or second surgery or the reaction on treatments by drugs. Especially for residual tumor tissue proliferation, markers are important because they may indicate the growth rate and the aggressiveness of the tumor. Radiation therapy is indicated in many of these recurrent tumors and can improve the prognosis. PMID- 16311412 TI - Treatment of pituitary tumors: surgery. AB - Following a century of technical developments and refinements, a variety of standard operation techniques to date are available for the surgical treatment of pituitary tumors. The vast majority of the lesions can be dealt with satisfactorily utilizing transsphenoidal approaches. The goal of surgical treatment is rapid eradication of the tumor mass, decompression of visual pathways, and elimination of hormonal oversecretion while preserving the normal gland and avoiding potential surgical complications. The tumor's size, extension, and configuration and the magnitude of hormonal oversecretion, are the essential factors that decide whether all the goals can be reached. Another important factor is the individual skill and experience of the surgeon. Still, several lesions that are mainly developed outside of the sella require transcranial approaches, of which the pterional and subfrontal routes are the most widely used. With microsurgical techniques and standard approaches, mortality is far below 1% and morbidity is remarkably low. The most favorable surgical results are obtained with microadenomas, which in the MR image are depicted as distinct low intensity lesions. Only recently has the recovery of pituitary function following surgery been convincingly demonstrated. With the extended transsphenoidal approaches, lesions become accessible that previously have been considered contraindications for transsphenoidal surgery. The introduction of new technical gadgets such as neuronavigation, endoscopy, and intraoperative imaging open new avenues and, even more, widen the spectrum of accessible lesions. Indications for surgery, the preoperative workup, surgical techniques, results, limitations, and new technical developments are briefly reviewed in this article. PMID- 16311413 TI - Treatment of pituitary tumors: radiation. AB - In this paper, the role of conventional radiotherapy and radiosurgery in the management of pituitary tumors is reviewed. After a short summary of the mechanism of action of irradiation therapy and the types of different irradiation techniques, the therapeutic effects and side effects are analyzed in patients with different types of pituitary tumors, including our own experience with conventional radiotherapy and radiosurgery in patients with acromegaly. Conventional fractionated radiotherapy has long been used to control growth and/or hormonal secretion of residual or recurrent pituitary tumors. However, patient selection for conventional radiotherapy still remains a controversial issue, because a number of potentially significant side effects, including hypopituitarism and other complications, have been described. Stereotactic radiotherapy/radiosurgery methods have several potential advantages over conventional radiotherapy, including their use in patients with residual or recurrent pituitary tumors who had previously been treated by conventional radiotherapy, but long-term follow-up data with these relatively new techniques are still limited. PMID- 16311414 TI - Gamma knife radiosurgery for pituitary adenomas. AB - Although surgical extirpation by transsphenoidal microsurgery is a major remedy for pituitary adenomas, adjuvant therapy also plays an important role in achieving tumor growth control and endocrine normalization in hormone-secreting tumors. Historically, the treatment options for pituitary adenomas included microsurgery, medical management, and fractionated radiotherapy, but radiosurgery has recently emerged as a practical treatment option. In this paper, we will describe the indications, radiosurgical procedure, results, histological change, and complications of gamma knife radiosurgery (GKS) for pituitary adenomas based on our experience since 1991 and a review of the literature. PMID- 16311415 TI - Treatment of pituitary tumors: somatostatin. AB - Somatostatin is an important physiological regulator of neuroendocrine function across multiple biological systems, including the brain and the gastrointestinal tract. In the pituitary gland, somatostatin regulates the secretion of hormones such as growth hormone and thyroid-stimulating hormone in healthy and pathological states. The short half-life of somatostatin makes it unsuitable for clinical use in chronic diseases, which led to the development of long-acting somatostatin analogs for the treatment of acromegaly and thyroid-stimulating hormone-secreting adenomas, which were administered by intermittent injection twice or three times a day. More recently, depot versions have been developed that permit dosing once every month. This review assesses the efficacy of somatostatin analogs in the treatment of pituitary adenomas, including acromegaly, thyroid-stimulating hormone-secreting tumors, non-functioning adenomas, and Cushing's disease. PMID- 16311416 TI - Treatment of pituitary tumors: dopamine agonists. AB - The neurotransmitter/neuromodulator dopamine plays an important role in both the central nervous system and the periphery. In the hypothalamopituitary system its function is a dominant and tonic inhibitory regulation of pituitary hormone secretion including prolactin- and proopiomelanocortin-derived hormones. It is well known that dopamine agonists, such as bromocriptine, pergolide, quinagolide, cabergoline, and lisuride, can inhibit PRL secretion by binding to the D(2) dopamine receptors located on normal as well as tumorous pituitary cells. Moreover, they can effectively decrease excessive PRL secretion as well as the size of the tumor in patients having prolactinoma. Furthermore, dopamine agonists can also be used in other pituitary tumors. The major requirement for its use is that the tumor cells should express D(2) receptors. Therefore, in addition to prolactinomas, targets of dopamine agonist therapy are somatotroph tumors, nonfunctioning pituitary tumors, corticotroph pituitary tumors, Nelson's syndrome, gonadotropinomas, and thyrotropin-secreting pituitary tumors. It is also an option for the treatment of pituitary disease during pregnancy. Differences between the effectiveness and the resistance of different dopaminergic agents as well as the future perspectives of them in the therapy of pituitary tumors are discussed. PMID- 16311417 TI - Treatment of pituitary tumors: pegvisomant. AB - Pegvisomant is a pegylated analog of growth that functions as a growth hormone receptor antagonist. The drug is capable of normalizing serum IGF-I concentrations (the chief mediator of disease activity in acromegaly) in 97% of patients, and therapy is associated with significant improvements in the symptoms and signs of GH excess. Biochemical control may be achieved with pegvisomant in patients wholly or partially resistant to somatostatin analogs, and there are emerging data to suggest that the drug may be particularly suitable for patients with acromegaly and co-existent diabetes mellitus. PMID- 16311418 TI - Pituitary carcinoma: diagnosis and treatment. AB - Pituitary carcinomas are rare pituitary tumors that by definition have cerebrospinal and/or systemic metastases. Most of the tumors occur in the setting of multiple recurrences of invasive pituitary adenomas. This article reviews the clinical presentation of these tumors, their neuroimaging and pathological features, tumor pathogenesis, and possible treatment modalities. PMID- 16311419 TI - Treatment of acromegaly: future. AB - Recent progress in the therapy of GH-secreting pituitary tumors includes three treatment modalities: surgery, radiotherapy, and medications. A combination of treatment options is often required to attain therapeutic goals, increasing the potential for a combination of unwanted side effects. The focus of this review is to discuss medical therapy of GH-secreting adenomas focusing on newer drug compounds. In selected cases, therapeutic goals are attained with somatostatin analog treatment alone. The GH receptor antagonist controls IGF-I hypersecretion, and its use in combination with somatostatin analogs in selected patients is tempting but requires further evaluation. Somatostatin multireceptor ligand SOM230 and a somatostatin-dopamine chimeric ligand are new compounds that may improve therapy outcome. Careful individualization of therapy is important in deciding the ideal treatment approach, and primary medical therapy may be recommended in selected patients. PMID- 16311420 TI - Precision assessment and radiation safety for dual-energy X-ray absorptiometry: position paper of the International Society for Clinical Densitometry. AB - Measurement of bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA) is used to diagnose osteoporosis, assess the risk of fracture, and monitor changes in BMD over time. Because biological changes in BMD are usually small in proportion to the error inherent in the test itself, interpretation of serial BMD tests depends on knowledge of the smallest change in BMD that is beyond the range of error. This value, called the least significant change (LSC), varies according to the instrument used, the patient population being tested, the measurement site, the skill of the technologist at positioning the patient and analyzing the test, and the confidence interval used in the calculation. The precision and LSC values provided by the manufacturer cannot be applied to clinical bone densitometry centers because of the differences in the patients being tested and the technologist performing the test. Because harmful errors in clinical management may occur from incorrectly interpreting serial BMD tests, it is recommended that every DXA technologist conduct a precision assessment and calculate the LSC for each measurement site and DXA instrument used. Precision assessment provides direct benefit to patients by allowing clinicians to make clinical decisions based on genuine change or stability of BMD. The patient-care benefits of precision assessment outweigh the risk of exposure to trivial doses of ionizing radiation. PMID- 16311421 TI - Comparison of dual-energy X-ray absorptiometry and conventional radiography for the detection of vertebral fractures. AB - Prevalent vertebral fractures confer a high risk of subsequent fractures; therefore, fracture identification is a key issue in osteoporosis management. Dual-energy X-ray absorptiometry has become the most widespread technique for the assessment of bone density, and can also provide images for assessment of vertebral morphometry (MXA) that are undistorted by parallax. The radiation dose is very low compared with conventional radiography. Whether the resolution of MXA is adequate for vertebral morphometry is uncertain. The present study compares MXA with conventional radiography in detecting vertebral deformities. MXA scans and lateral radiographs were performed in 61 women with osteoporosis. Adequate images were obtained in 99.1% of the 793 vertebrae by radiography and 95.6% by MXA. Poor image quality was mostly at T4-6 by MXA. Precision of repeat analyses was comparable between the techniques. Heights measured by MXA were 24 to 35% less than the radiographic values, the methods produced equivalent results for the ratio of anterior to posterior heights, but the MXA values for middle to posterior ratios were about 10% lower than for the radiographs (p < 0.0001). The extent of agreement between the techniques in identifying vertebral fractures was dependent on the strictness of the fracture definition used. With a 3 standard deviation (SD) criterion, the techniques found similar numbers of patients to have vertebral deformities, and the concordance in classification of individual vertebrae was 94%. At a 2 SD, cut-off concordance was 79%, and at 4 SD, it was 96%. Receiver-operating characteristic curves using cut-offs of > or = 3 SDs also showed excellent diagnostic accuracy. We conclude that MXA shows acceptable performance for clinical use in diagnosing vertebral deformities, as long as cut offs of > or = 3 SDs are used, although a few percent of vertebrae in the upper thoracic region cannot be imaged adequately using this technique. PMID- 16311422 TI - Prospective identification of postmenopausal osteoporotic women at high vertebral fracture risk by radiography, bone densitometry, quantitative ultrasound, and laboratory findings: results from the PIOS study. AB - Women with established osteoporosis are at high risk to sustain additional vertebral fractures. Treatment may affect the predictive power of bone densitometry and biochemical techniques. There are few prospective studies comparing fracture prediction by dual-energy X-ray absorptiometry (DXA) and other techniques in treated women with established osteoporosis. The objective of this study was to prospectively assess the predictive power of various DXA and quantitative ultrasound (QUS) techniques for identification of women at high risk to develop new fractures over 1-2 yr. Moreover, we wanted to investigate whether previous or ongoing therapy precluded the use of common clinical laboratory blood tests and bone turnover markers for prediction of fracture risk. We measured prevalent fracture status; bone mineral density (BMD) of the whole body, spine, and hip by DXA; QUS of the calcaneus and the patella; hormones and various markers of bone resorption and formation; and took standard blood tests in 124 women (age 64.9 yr +/- 7.9) with manifest and variously treated postmenopausal osteoporosis. Subsequently, new spine fractures were assessed after 1 yr and, in a subset of 87 women, after 2 yr. Prevalent fractures turned out to be the strongest predictor of subsequent vertebral fractures with an age-adjusted odds ratio (OR) of 3.9 per prevalent fracture over 2 yr. Furthermore, our results underline the predictive power of spinal BMD (sOR = 2.1; standardized OR per 1 standard deviation population variance decrease), whole body BMD (sOR: 2.4), and QUS stiffness index of the calcaneus (sOR: 2.8) for vertebral fracture prediction. QUS of the patella did not predict vertebral fractures. Blood sedimentation rate was predictive in the first year (sOR: 1.9). The predictive power of bone turnover markers, however, appeared to be too low to be detectable in a group of this sample size and it may have been reduced because most women were already receiving treatment. In conclusion, radiographic measures, but not the tested laboratory bone turnover markers, enabled us to identify women (from a population of osteoporotic women who have been treated for some time with a variety of medications) who are at highest risk for developing new vertebral fractures within 1-2 yr. PMID- 16311423 TI - Relationship among MRTA, DXA, and QUS revisited. AB - Inexpensive, commercially produced devices that directly measure bone strength in vivo are not currently available. Mechanical response tissue analysis (MRTA), a unique prototype device, is an in vivo vibrational test that measures transverse bending stiffness (a measure of whole bone strength expressed as the product of estimated Young's modulus of elasticity and cross-sectional moment of inertia, EI, Nm2) at ulna midshaft. We compared speed of sound (SOS; [m/s]) in ulna cortical bone using a commercially available axial transmission quantitative ultrasound (QUS) device with EI using MRTA. Dual-energy X-ray absorptiometry (DXA) was used to provide an estimate of ulna size (cm2), bone mineral content (BMC; [g/cm]) and areal bone mineral density (BMD; [g/cm2]). The objective of the study was to determine if ulna SOS--alone or in combination with BMD from DXA- was correlated with ulna EI, thus becoming a surrogate measure of transverse bending stiffness, and thus whole bone strength. Data were collected from 138 female volunteers (18-86 yr). EI and SOS were significantly correlated, r = +0.218, p = 0.01, but r2 was very low, 4.8%. SOS and total ulna BMD were combined to estimate elastic modulus, which correlated with EI, r = +0.377, p < 0.0001; however, the correlation was not significantly better than with SOS alone. We conclude that axial transmission QUS is not a strong surrogate in vivo technique for estimating transverse bending stiffness. PMID- 16311424 TI - Prevalence of osteoporosis using DXA bone mineral density measurements at the calcaneus: cut-off points of diagnosis and exclusion of osteoporosis. AB - The objective of this article is to evaluate different T-score cut-off points in the calcaneus in order to establish the prevalence of osteoporosis in the general population and to evaluate the clinical value of bone mineral density at the calcaneus as a tool to identify patients with spine or hip osteoporosis. A total of 1455 people (727 women and 728 men) from the Hortega cohort were studied. The densitometric studies were carried out in the calcaneal region using a Peripheral Instantaneous X-ray Imaging (PIXI) Lunar bone densitometer. We established three cut-off points (-1.6, -2.0, -2.5). One-hundred twenty-five patients (67 men with a mean age of 47 +/- 13 yr and 58 women with a mean age of 66 +/- 8 yr) from internal medicine outpatient clinics who were undergoing densitometry of the calcaneus, spine, and hip were subsequently studied. The prevalence of osteoporosis in women with a calcaneus T-score -1.6 was 12.4%, which is comparable to the 12.7% obtained with an axial densitometer in a Spanish population. The prevalence in men was 7.8%, with a calcaneus T-score of <-2.0. In women, the highest sensitivity (85%) was obtained with a calcaneus T-score of < 1.0 and the highest specificity with a calcaneus T-score of <-2.5. In men, the best sensitivity (61%) was obtained with a calcaneus T-score of <-1.0 and the best specificity (98%) with a calcaneus T-score of <-2.5. A calcaneus T-score < 1.6 is an adequate cut-off to establish the prevalence of osteoporosis in population studies. In women, a calcaneus T-score of >-1.0 enables the diagnosis of the disease to be excluded, whereas a calcaneus T-score of <-2.5 enables the diagnosis to be confirmed and treatment to be initiated in both men and women. PMID- 16311425 TI - Ethnic differences in bone mineral density between inuit and Caucasians in north Greenland are caused by differences in body size. AB - Data on bone mineral density (BMD) in living Inuit are limited and BMD measurements in Arctic Inuit using Dualenergy X-ray Absorptiometry (DXA) are lacking. Ethnicity may be important for bone mass. The aim of this study was to validate DXA in rural Arctic Greenland, to measure BMD in Greenland Inuit and Caucasians, and to estimate the importance of ethnicity for BMD. We measured the BMD in 80 healthy subjects living in Ilulissat and Saqqaq in North Greenland twice in both distal forearms and in both heels using peripheral DXA (pDXA). Participants were stratified by origin (Inuit[settlement])/Caucasians, n = 33 [19]/28), gender (men/women, n = 37/43), and age (30-39/40-49, n = 32/48). Caucasians were bigger than Inuit (men/women, height p < 0.001/p < 0.001; weight p = 0.01/ p = 0.026), but had similar BMI (p = 0.42/0.70). Triplicate pDXA measurements showed individual CV% = 0.16-1.79%; overall CV% = 1.1% (forearm)/1.0% (heel). Data followed the normal distribution (p = 0.65-0.99) with identical variances between Inuit and Caucasians (p = 0.12-0.63). Mean BMD in right forearm/left forearm/right heel/left heel was: Inuit men 0.570/0.568/0.549/0.536 g/cm2; Inuit women 0.484/0.474/0.473/0.464 g/cm2; Caucasian men 0.580/0.570/0.646/0.638 g/cm2; Caucasian women 0.495/0.496/0.552/0.553 g/cm2. An ethnic difference in heel BMD (p < 0.001) disappeared when adjusted for weight (p = 0.30). No difference was found in forearm BMD. In conclusion, pDXA is feasible and reliable in rural Greenland. Ethnic differences in BMD are small and may reflect differences in body size. PMID- 16311426 TI - Effective dose of dual-energy X-ray absorptiometry scans in children as a function of age. AB - Effective dose, a parameter utilized to assess biological risk related to radiation exposure, may be used to evaluate risk associated with dual-energy X ray absorptiometry (DXA). We estimated the effective dose from DXA (Hologic QDR 4500A) scans of the lumbar spine (fast array mode), total body, hip (fast array mode), and forearm for children ages 1, 5, 10, and 15 yr and for adults. Entrance dose incorporating backscatter was determined for each scan type. Depth-dose curves were derived using Plexiglas slabs simulating tissue attenuation. Organ depth was estimated using pediatric phantom models. For all scan types, the effective dose decreased as age increased. The effective dose values for a 1-yr old and an adult, respectively, were 4.7 microSv and 2.2 microSv for a lumbar spine scan performed in fast array mode, 3.4/3.5 microSv and 1.8/2.1 microSv (male/female) for a total body scan, and 0.14 microSv and 0.03 microSv for a forearm scan. There were marked sex differences in the effective dose associated with hip scans (fast array mode) ranging from 15.2 microSv for a 1-yr-old male to 4.6 microSv for an adult female. A comprehensive uncertainty analysis indicated that the effective dose values were reliable within a factor of 3. With the exception of the hip scans in 1- and 5-yr-olds, the effective doses were below the negligible individual dose limit of 10 microSv/yr. PMID- 16311427 TI - Forearm single X-ray absorptiometry in the identification of postmenopausal women with osteoporosis at the hip and spine: a correlation study. AB - The International Society for Clinical Densitometry (ISCD) has stated that forearm bone mineral density (BMD) testing combined with a thorough clinical evaluation may be an option for the diagnosis of osteoporosis when central bone density (CBD) testing is not available. This study assessed the performance of two different forearm sites in identifying subjects with spinal and femoral osteoporosis, and defined the 90% sensitivity point for the DTX-100 bone densitometer in the detection of central osteoporosis. Four hundred and two postmenopausal Bulgarian women between the ages of 50 and 81 yr (mean age 60.24 +/- 10.48 yr) participated in this study. Forearm BMD (distal and ultradistal forearm) was measured with a DTX-100 device (Osteometer Meditech, USA) and central BMD (lumbar spine and proximal femur) with a Hologic QDR 4500 A device. Linear T-score correlations among sites, sensitivity and specificity of the forearm site were analyzed. T-score correlations between the forearm and the central sites ranged from 0.32 to 0.69 (p < or = 0.05 for all correlations in age group 50-59). The forearm site sensitivity increased slightly with advancing age, but specificity decreased. When the distal forearm BMD cut point (0.340 g/cm2) was set to achieve 90% sensitivity to identify total hip osteoporosis, specificity was 40%; when the distal forearm BMD cut point (0.410 g/cm2) was set to achieve 90% sensitivity to identify spinal osteoporosis, specificity was 55.4%; when ultradistal forearm BMD cut points (0.280 and 0.320 g/cm2) were set to achieve 90% sensitivity to identify total hip and spinal osteoporosis, specificity was 40.8 and 59.2%, respectively. Forearm bone density measures may be useful to selectively screen for patients with central osteoporosis. PMID- 16311428 TI - Postmenopausal women with colles' fracture have lower values of bone mineral density than controls as measured by quantitative ultrasound and densitometry. AB - Measurement of ultrasonographic parameters provides information concerning not only bone density but also bone architecture. We investigated the usefulness of ultrasonographic parameters and bone mineral density (BMD) to evaluate the probability of Colles' fracture. Two-hundred eighty-nine postmenopausal women (62.3 +/- 8.7 yr) with (n = 76) and without (n = 213) Colles' fracture were studied. BMD of lumbar spine and proximal femur was evaluated in all women by dual-energy X-ray absorptiometry (DXA) and speed of sound (SOS), broadband ultrasound attenuation (BUA), and stiffness in the calcaneus were measured by a Sahara ultrasonometer (Hologic). Patients suffering from Colles' fracture had lower values of BMD adjusted by height at the lumbar spine, L2-L4 (0.797 g/cm2 vs 0.860 g/cm2), femoral neck (0.685 g/cm2 vs 0.712 g/cm2 ), SOS (1518 m/sg vs 1525 m/sg), and stiffness (74.6 vs 77.7) (p < 0.05). Nevertheless, BUA values were similar in both groups. After stepwise logistic regression analysis, the area found under receiver operating characteristic (ROC) curves was 0.60 for L2L4 and 0.63 for a formula combining L2L4 and height. Our data suggest that patients suffering from Colles' fracture have lower values of BMD by DXA, SOS, and stiffness. However, the ability of these techniques to discriminate is low because the values for the area under ROC curve are 0.60 for L2-L4 and 0.63 for a formula derived of the combination of L2-L4 and height. PMID- 16311429 TI - Effect of temperature on the longitudinal variability of quantitative ultrasound variables. AB - It is unclear whether longitudinal change in phantom measurements bears any relation to the long-term in vivo instrument performance of quantitative ultrasound devices. Longitudinal quantitative ultrasound phantom data were obtained by measuring the manufacturer-provided phantom at ambient temperature and two different sets of Leeds phantoms at either ambient temperature or following a phantom temperature-control protocol. Measurements were performed using the Achilles Plus bone densitometer. Changes in longitudinal phantom data were compared to in vivo quantitative ultrasound data obtained from seven healthy, young volunteers. A cosinor model with linear trend and Hotelling's T2 test were used to quantify seasonal rhythms and long-term drift in quantitative ultrasound variables. Temperature effects and marked seasonal rhythms on quantitative ultrasound phantom measurements were evident but were far less apparent in vivo. Longitudinal precision of quantitative ultrasound variables was poorer for the manufacturer-provided phantom than for phantoms that were subjected to a temperature-control protocol or for healthy volunteers. This study has shown that longitudinal precision and longitudinal change differs between in vivo and phantom data. Longitudinal quantitative ultrasound measurements for monitoring change in skeletal status cannot, as yet, be properly controlled. PMID- 16311430 TI - Bone assessment in children: comparison of fan-beam DXA analysis. AB - The newest version of whole body dual-energy X-ray absorptiometry (DXA) analysis software from Hologic (Discovery 12.1) is designed to enhance bone detection in smaller subjects. We re-analyzed 1127 pediatric scans (ages 1.8-18.5 yr) previously analyzed using software version 11.2. Regression analysis compared new and original results for bone area (BA), bone mineral content (BMC), bone mineral density (BMD), and DXA-derived body weight. Changes in total and regional bone results were compared with age, weight, and height. New results were highly correlated with original analyses (R2 > or = 0.9), but there were large differences at the individual subject level. The BA and BMC values increased in subjects less than 40 kg weight, resulting in a lower BMD. Original and new results were equivalent by about age 14 yr in both genders. Regional bone data showed the greatest changes in the legs. The newest software produces significant changes in bone values in subjects weighing less than 40 kg, compared with earlier versions. This effect increases with decreasing body weight. This will impact interpretation of longitudinal pediatric DXA studies, as well as existing pediatric whole body bone reference databases. Investigators must recognize which DXAsoftware version they are using, and which version produced any reference database they may use for comparison. PMID- 16311431 TI - Low 25-hydroxyvitamin D and osteopenia are prevalent in persons > or =55 yr with fracture at any site: a prospective, observational study of persons fracturing in the community. AB - Ethnic diversity and lower socioeconomic populations are poorly captured in most studies of osteoporosis and fracture risk. This article describes a prospective, observational study designed to analyze risk factors for fracture in an ambulatory, ethnically diverse, urban population aged > or =55 yr. The goal of the study was to determine the number of fractures associated with hypovitaminosis D (< or =15 ng/mL serum 25-hydroxyvitamin D) and osteopenia (T score <-1.5) by bone mineral density (BMD). From January 1 to July 31, 2001, we identified 262 persons who fractured in our community; 83 chose to enroll in the study. Enrolled patients had a BMD examination at two sites; their blood was drawn for 25-hydroxyvitamin D (25VitD), calcium, phosphorus, albumin, and alkaline phosphatase. At the completion of the study a letter was sent to the patients detailing the findings, and a copy sent to their physician. Of the 83 persons enrolled, 73 (88%) had evidence of osteopenia or osteoporosis (T-score < 1.5) and/or low 25VitD. All fractures in the community in person > or =55 yr, with or without a history of antecedent trauma, should be assessed with BMD and screening for 25VitD. PMID- 16311432 TI - High degree of discordance between three-dimensional and two-dimensional lumbar spine bone mineral density in Turner's syndrome. AB - Low bone mineral density (BMD) measured by dual-energy X-ray absorptiometry (DXA) has been described in Turner's syndrome (TS). One of the error factors of DXA is short stature, a common finding in TS patients. Aimed to evaluate the influence of a low stature on BMD, we compared the two-dimensional (2D) or conventional BMD (cBMD) with three-dimensional (3D) or volumetric BMD (vBMD) in 62 females (10 to 48 yr old) with TS diagnosis in a case control study. They were compared to 102 normal females (7 to 45 yr old) grouped by age-ranges. All patients were subjected to a lumbar spine densitometry by DXA in the PA and lateral projections, obtained the cBMD and vBMD and calculated for the apparent BMD (appBMD). In TS, the mean of Z-score for cBMD was significantly lower than that for vBMD and for appBMD (-2.31 +/- 1.42; -0.64 +/- 1.55; and -1.72 +/- 1.5; respectively). Most of the patients (83.8%) had a Z-score <-1 for cBMD, whereas the majority (58.1%) had a Z-score <-1 for vBMD. Concluding, the cBMD underestimates the bone mass of the lumbar spine in patients with TS inducing to false diagnoses of bone fragility. Volumetric BMD approached the bone mass of control patients, while appBMD just partially do that. PMID- 16311433 TI - Does the crack silently break the back? Long-term follow-up and assessment of risk factors for regional osteoporosis following tibial shaft fractures. AB - The aim of this study is to determine the incidence and to quantify the risk factors for developing long-term regional osteopenia/osteoporosis (RO) following tibial fractures. We studied 42 adult subjects (8 females and 34 males) who had sustained a tibial fracture 16 yr prior to the study. Mechanism and type of injury, method of treatment, length of immobilization, weight-bearing status, and healing time were determined from the patient records. A questionnaire covering known causes of generalized osteoporosis (GO), including history of smoking, alcohol consumption, medications, other fractures, thyroid/parathyroid disorders, epilepsy, and renal disorders, was answered by all the subjects. Bone mineral density (BMD) of lumbar vertebrae 1-4 and both proximal femurs was assessed using dual-energy X-ray absorptiometry (DXA) scanning. T- and Z-scores were generated. Assessment of risk factors was done by calculating the odds ratio (OR) and 95% confidence interval (CI). The incidence of significant loss of BMD as defined by the World Health Organization (T-score <-1) of the ipsilateral neck of femur and/or lumbar spine was found to be 40%. None of our subjects had any known cause for GO. The following risk factors were found to be statistically significant in unadjusted models: smoking (OR 22, 95% CI = 4 - >40), alcohol >20 units/wk (OR 11, 95% CI 2 = 2-54), open fracture (OR 17, 95% CI = 2.9 - >40), nonweight bearing >4 wk (OR 15, 95% CI 2.9- >40), and delayed union defined as healing time more than 6 mo (OR 15, 95% CI 1.54 - >40). Permanent regional loss of BMD occurs in a significant proportion of tibial shaft fracture patients. Modern fracture management should include identifying "at-risk" patients and appropriate preventive measures to prevent fragility fractures. PMID- 16311434 TI - Evaluation of femur angle abduction/adduction and bone mineral density values. AB - Bone densitometry using fan-beam technology allows both femurs to be rapidly measured with the aid of a bilateral foot-positioning tool to keep both legs in internal rotation. The Lunar Prodigy system uses a bilateral foot-positioning tool with a fixed interpedal width. The aim of this study was to determine whether the small degrees of femur abduction and adduction introduced by a fixed interpedal width-positioning tool influences the femur bone mineral density (BMD) measurements. Two-hundred patients had the total femur (FTot), femoral neck (FN), femoral shaft (FS), and femoral trochanter (FT) BMD, and angles of femoral abduction and adduction measured. Of all subjects, 26.5% had an absolute difference in femur angulation of 1 degrees , 77.5% had 5 degrees and 97% had 10 degrees. There was no significant correlation between the right-to-left differences in femur angles and the absolute right-to-left differences in FTot, FN, FS, and FT BMD. The introduction of small degrees of femur angulation with the fixed interpedal width bilateral foot positioner does not significantly affect the measured BMD at the various femur regions. PMID- 16311435 TI - DXA scanning of acetabulum in patients with cementless total hip arthroplasty. AB - The aim of this study was to evaluate the reproducibility of bone mineral density (BMD) measurements of the periprosthetic bone in patients with hemispherical acetabular cups in cementless total hip arthroplasty (THA). Thirty patients were treated for primary osteoarthrosis with cementless THA. Dual-energy X-ray absorptiometry (DXA) scanning was performed with a pencil-beam bone densitometer (Norland XR-36). Accuracy and reproducibility was determined by double measurements of BMD in four regions of interest (ROI). The influence of patient postures including various pelvic inclination angles was evaluated as well. Pitman test for a combined netROI revealed a standard deviation ratio of 3.2 for the anterio-posterior scans related to the lateral position. The Wilkinson ROIs showed a high intraobserver agreement. With pelvic tilt increasing until 20 degrees , the precision of DXA scanning decreased. In conclusion, reproducibility of DXA scanning was high. This study demonstrated that the patients can be scanned in the supine position, and BMD measurement of the periacetabular bone can be performed using the Wilkinson model with four rectangular ROIs. PMID- 16311436 TI - Patients with knee osteoarthritis have lower total hip bone mineral density in the symptomatic leg than in the contralateral hip. AB - Osteoarthritis (OA) of the knee is common in the aging population. In patients with OA, bone mineral density (BMD) is usually increased, but the fracture rate does not appear to be systematically lower than in age-matched healthy controls. The aim of our study was to describe hip BMD in patients presenting with unilateral symptomatic knee OA. Patients with painful knee OA were prospectively included in a single-center, randomized, double-blind, placebo-controlled clinical trial to evaluate the structure-modifying efficacy of an oral chondroitin sulfate treatment on the knee joint. The majority of these patients underwent additional measurements of BMD of their lumbar spine and both hips using dual-energy X-ray absorptiometry (DXA). The hip BMD values of the leg with symptomatic knee OA were compared with the contralateral hip. One-hundred and sixty-one patients (81 men and 80 women; aged 62.6 +/- 9.2 yr, range 40-82 yr) underwent DXA. The median total hip BMD was higher than in age-matched controls, but patients had a relatively lower hip BMD in the knee OA-affected leg (p = 0.001). Our knee OA patients rarely presented with concomitant osteoporosis, but usually had a relatively lower hip BMD on the affected leg. Therefore, we suggest that the hip of the leg with symptomatic knee OA should be measured if DXA is acquired only at one hip. Future studies have to assess whether the relative decrease of BMD at the hip of the leg with knee OA might influence fracture incidence. PMID- 16311437 TI - Improved performance of hip DXA using a novel region of interest in the upper part of the femoral neck: in vitro study using bone strength as a standard of reference. AB - We tested the hypothesis that bone mineral density (BMD) and bone mineral content (BMC) in proximal human femur specimens in the upper neck region of interest (ROI) and femoral neck axis length (FNAL) provide a significantly better prediction of femoral bone strength than standard ROIs in vitro. BMD and BMC were measured in 110 proximal femur specimens using a standard dual-energy X-ray absorptiometry (DXA) scanner. The analysis included a new ROI in the upper neck as well as the standard ROIs. FNAL was obtained from the scan images. The specimens' failure-load was measured in a mechanical loading device, simulating a fall on the greater trochanter. For the standard ROIs, correlations between failure-load and BMD ranged from R2 = 0.64 (shaft ROI) to R2 = 0.70, p < 0.001 (femoral neck). Prediction of strength by BMD did not significantly differ from those of BMC (R2 ranging from 0.65 to 0.75, p < 0.001). In the upper neck ROI, for both BMD and BMC correlations with failure-load were higher (R2 = 0.76 and 0.81, respectively; p < 0.001). A lower, yet still significant, correlation was found between FNAL and bone strength (R2 = 0.23, p < 0.001). Normalization of failure-load with respect to FNAL did not significantly increase the correlations with densitometric measures. This study provides in vitro evidence indicating that among the ROIs of the proximal femur the newly defined upper neck ROI provides the best prediction of bone strength. Only a weak association was observed between failure load and FNAL. PMID- 16311438 TI - The arterial system pressure-volume loop. AB - Although the ventricular P-V loop has become a popular tool to characterize aspects of the performance of the heart, an arterial system P-V loop has not yet been described. In principle, the volume stored in the arterial system (V) could be calculated by integrating the difference between inflow and outflow. In practice, however, flow out of the innumerable arterioles cannot be measured directly. To overcome this obstacle, it has been shown that outflow can be approximated by input pressure divided by total peripheral resistance. Recently, the classical Windkessel model was generalized with the concept of apparent arterial compliance (C(app)), the transfer function relating pressure and volume expressed in the frequency domain. The arterial system P-V loop serves as a time domain representation of C(app). This simple technique provides the first known characterization of an arterial system P-V loop. PMID- 16311439 TI - Effects of the time response of the temperature sensor on thermodilution measurements. AB - Thermodilution is widely used to measure cardiac output, ejection fraction and end diastolic volume. Even though the method is based on dynamic temperature measurements, little attention has been paid to the characterization of the dynamic behavior of the temperature sensor and to its influence on the accuracy of the method. This paper presents several theoretical and empirical results related to the thermodilution method. The results show that, at flow velocities above 0.2 m s(-1), the response of temperature sensors embedded in Swan-Ganz catheters can be accurately described by a convolution operation between the true temperature of the blood and the impulse response of the sensor. The model developed is used to assess the influence of the probe response on the measurement of cardiac output, and this study leads us to the conclusion that the probe response can cause errors in the cardiac output measurement, but this error is usually small (2% in cases with a high degree of arrhythmia). The results show that these small errors appear during arrhythmias that affect the R-R interval and when the real temperature distribution at the pulmonary artery does not possess a shape with perfect temperature plateaux. PMID- 16311440 TI - Measuring the rate of change of haemodynamic response at the onset of exercise in normal limbs and those with intermittent claudication. AB - Patients with claudication have an inadequate haemodynamic response to exercise. Blood flow response will not only have a magnitude, but also a rate of change. There is scope for investigating these parameters, as manipulation of the factors which control them may benefit work to improve the treatment for claudication. This work compares the responses for patients with one normal limb and one with intermittent claudication. A custom-built ergometer allows unilateral, infragenicular plantar flexion exercise, whilst common femoral artery blood flow can be measured continuously by Duplex ultrasound. This apparatus was used to measure blood flow before, at the onset of and during a 5 W square-wave exercise stimulus in 15 patients. The claudicant group had a mean steady-state gain that was approximately half that of the normal group at around 170 ml min(-1) (p < 0.001) and a response time that was much shorter (p = 0.006). A mean response time of 21.0 +/- 1.4 s was achieved in claudicant limbs compared to 31.8 +/- 2.9 s in normals. However, the average rate of change of blood flow during this response time was estimated to still be greater for the normal group, at 431.7 +/ 47.1 ml min(-2), than for the claudicant group. The differences in magnitude and rate of change of limb blood flow response to exercise in claudicants were significant and may have implications for the treatment of claudication. PMID- 16311441 TI - Quantification of synchronization during atrial fibrillation by Shannon entropy: validation in patients and computer model of atrial arrhythmias. AB - Atrial fibrillation (AF), a cardiac arrhythmia classically described as completely desynchronized, is now known to show a certain amount of synchronized electrical activity. In the present work a new method for quantifying the level of synchronization of the electrical activity recorded in pairs of atrial sites during atrial fibrillation is presented. A synchronization index (Sy) was defined by quantifying the degree of complexity of the distribution of the time delays between sites by Shannon entropy estimation. The capability of Sy to discriminate different AF types in patients was assessed on a database of 60 pairs of endocardial recordings from a multipolar basket catheter. The analysis showed a progressive and significant decrease of Sy with increasing AF complexity classes as defined by Wells (AF type I Sy = 0.73 +/- 0.07, type II Sy = 0.56 +/- 0.07, type III Sy = 0.36 +/- 0.04, p < 0.001). The extension of Sy calculation to the whole right atrium showed the existence of spatial heterogeneities in the synchronization level. Moreover, experiments simulated by a computer model of atrial arrhythmias showed that propagation patterns with different complexity could be the basis of different synchronization levels found in patients. In conclusion the quantification of synchronization by Shannon entropy estimation of time delay dispersion may facilitate the identification of different propagation patterns associated with AF, thus enhancing our understanding of AF mechanisms and helping in its treatment. PMID- 16311442 TI - Spatiotemporal pattern of the extracranial component of the rheoencephalographic signal. AB - The use of rheoencephalography (REG) in the clinical practice to evaluate cerebral blood flow is conditional on the finding of a method for removing the extracranial interference caused by the scalp blood flow. To remove this undesirable influence, digital processing based on statistics could be an effective technique if the appropriate data model were applied. This paper focuses on the analysis of the spatiotemporal features of the extracranial REG component, by comparing its morphology and phase shift at several scalp sites. For this purpose, a numerical model of the scalp was employed to assess tissue impedance changes caused by the inflow of a stepwise blood pulse wave. These results were compared with the experimental impedance waveforms recorded on six pairs of adjacent electrodes. The correlation coefficients between each pair of impedance recordings of each subject were always greater than 0.942, showing a mean value of 0.986. This result suggests that the extracranial REG component can be considered as morphologically invariant. On the other hand, negligible phase shifts were observed when mean electrode distances, measured in the blood flow direction, were relatively small, although temporal corrections in the data model would be advisable for longer distances. PMID- 16311443 TI - A compact electric potential sensor array for the acquisition and reconstruction of the 7-lead electrocardiogram without electrical charge contact with the skin. AB - Conventional electrocardiogram (ECG) systems make use of separate electrical connections to the arms and legs. These use a 'long baseline' for the voltage reference potential which in the case of precordial ECG leads is provided using a Wilson central terminal (WCT) wiring configuration. The aims of this project were (a) to construct compact, non-invasive surface ECG sensor arrays which would operate without the need for a WCT reference, (b) to obtain high quality precordial ECGs showing fine differences in ECG detail between small adjacent areas of the chest and (c) to reconstruct, from a compact array of four sensors, ECGs which closely match to the conventional 7-lead ECG system, but without the need for multiple wires and long baselines. In this paper, we describe two sensor array configurations which have been constructed using electric potential sensors (EPSs). We show high quality precordial ECGs obtained from small areas of the surface of the chest and show the different angular vectors (leads) in the frontal cardiac plane constructed using signals from the array elements. We suggest that these ECG arrays, which are simple to apply, should prove to be a valuable tool in providing useful information about the state of the heart. PMID- 16311444 TI - Can mechanical myotonometry or electromyography be used for the prediction of intramuscular pressure? AB - The aim of the study was to characterize the electromechanical properties of skeletal muscle during isometric loading as well as to assess the potential of estimating intramuscular pressure by electrical and mechanical methods. Simultaneous electromyography (EMG), mechanical myotonometry (MYO, frequency and decrement of decay) and intramuscular pressure (IMP) measurements were conducted at rest and during short-term and long-term isometric contractions in patients with chronic pain in the anterior leg or dorsal forearm. The EMG amplitude and MYO(freq) accounted significantly (24-73%, p < 0.0001) for the variations in the IMP under short-term isometric loading. The IMP, EMG and MYO(freq) increased linearly with the relative muscle load (r = 0.868-0.993, p < 0.05). Mean values of EMG amplitudes at the contraction levels of 75% and 100% maximum voluntary contraction (MVC) and MYO(freq) values at all contraction levels (0-100% MVC) were higher for subjects with pathological values of IMP than for those with IMP values in the normal range. Total changes in IMP and EMG amplitude during 1 min isometric contraction were linearly interrelated (r = 0.747, p < 0.0001). We conclude that both surface electromyography and myotonometry parameters are indicative of intramuscular pressure, but neither of these methods can be used alone to diagnose non-invasively chronic compartment syndrome with acceptable accuracy. PMID- 16311445 TI - Physiologically precise simulation of multiple lung gas exchange during anaesthesia by simultaneous gas infusion and extraction. AB - A lung gas exchange simulator was tested which produces simultaneous uptake and/or elimination of multiple gases by an artificial test lung with physiologically realistic gas expired and exhaust gas flows, using a combination of infusion of diluting/enriching gases into the lung with lung gas extraction. A deterministic algorithm is incorporated which calculates required gas infusion and extraction flow rates for any set of possible target gas exchange values with any given set of fresh gas flows and concentrations. Six different scenarios were simulated, comprising a range of gas exchange values for each gas species which lie within a physiologically realistic range for anaesthetized patients. For each of these experiments the system was tested for 15 consecutive measurements over 25 min by measurement of gas exchange in the system using the Haldane transformation. RESULTS: the mean bias and standard error of the mean bias (SE, in parentheses) relative to the target value was: +0.001 (0.002) l min(-1) for O(2) uptake, -0.002 (0.005) l min(-1) for CO(2) production, -0.001 (0.002) l min( 1) for uptake of nitrous oxide and +0.3 (0.1) ml min(-1) for uptake of a volatile anaesthetic agent (isoflurane). The confidence limits of the mean bias were within 5% of the target value for all gases and scenarios with the exception of those where a low uptake of anaesthetic gas was specified. The confidence limits of the mean bias for the lower uptakes of isoflurane were within 10% of the target value for these scenarios and within 15% for the low uptake of N(2)O. Good accuracy and precision of this approach to lung gas exchange simulation were demonstrated, resulting in a versatile simulator. PMID- 16311446 TI - Heart rate variability: comparison among devices with different temporal resolutions. AB - Several devices based on ECG can acquire beat-to-beat intervals, and some of these devices may be used for heart rate variability (HRV) analysis. Most of these devices and their methods to acquire the beat-to-beat intervals need to be validated for HRV analysis by comparing them against traditional methods. Some of these methods have low temporal resolution, which may be sufficient for certain studies. HRV analysis parameters obtained from two methods differing in temporal resolution were compared to the HRV analysis parameters obtained from a referent method (ECG). One of the methods had a high resolution (<1 ms) and the other method had a low resolution (10 ms). Seven healthy male volunteers participated in the study. The beat-to-beat intervals were collected simultaneously from the three methods and the HRV parameters derived from them were used for our comparisons. The Pearson product-moment correlations were used, which demonstrated an excellent correlation (r > 0.99) in time domain, frequency domain and some non-linear HRV measures. The HRV measures were further analyzed using the one-way repeated-measures analysis of variance (ANOVA) and statistically significant differences were observed in some of the HRV parameters obtained from the low-resolution method, especially the mean RR (p < 0.001) and the mean heart rate (p < 0.001). Since the same software was used for HRV analysis of the three methods, any differences were due to the temporal resolution of the RR intervals measured by the methods. Overall, both the methods correlated very well with the referent method among healthy volunteers in resting conditions and may be used by researchers for HRV studies. PMID- 16311447 TI - Subtraction of 16.67 Hz railroad net interference from the electrocardiogram: application for automatic external defibrillators. AB - The widespread application of automatic external defibrillators (AEDs) for treating out-of-hospital cardiac arrest incidents and their particular use at railway stations defines the task for 16.67 Hz power line interference elimination from the electrocardiogram (ECG). Although this problem exists only in five European countries, it has to be solved in all AEDs, which must comply with the European standard for medical equipment requirements for interchangeability and compatibility between rail systems. The elimination of the railroad interference requires a specific approach, since its frequency band overlaps with a significant part of the frequencies in the QRS spectra. We present a method based only on one channel ECG signal processing, which effectively subtracts the interference components. The computation procedure is based on simple signal processing tools, which include: (i) bi-directional band pass filtering (13-23 Hz) of the analyzed ECG segment; (ii) estimation of adequate linearity thresholds; (iii) frequency measurement and calculation of dynamic interference buffer in linear segments and (iv) phase synchronization and subtraction technique in nonlinear segments. The developed method has proved advantageous in providing sufficient quality of the output interference free ECG signal for seven defined arrhythmia types (normal sinus rhythm, normal rhythm, supraventricular tachicardia, slow/rapid ventricular tachycardia, and coarse/fine ventricular fibrillation), and simulated interferences with constant or variable frequencies and amplitudes, which cover the entire amplitude range of the input channel. The procedure is suitable to be embedded in AEDs as a preprocessing branch, which proves reliable for analysis of ECG signals, thus guaranteeing the specified accuracy of the AED automatic rhythm analysis algorithms. PMID- 16311448 TI - Accessing heart dynamics to estimate durations of heart sounds. AB - Segmentation of the phonocardiogram into its major sound components is the first step in the automated diagnosis of cardiac abnormalities. Almost all of the existing phonocardiogram segmentation algorithms utilize absolute amplitude or frequency characteristics of heart sounds, which vary from one cardiac cycle to the other and across different patients. The objective of this work is to provide an efficient phonocardiogram segmentation technique, under difficult recording situations, by utilizing the underlying complexity of the dynamical system (heart) giving rise to the heart sound. Complexity-based segmentation is invariant to amplitude and frequency variations of the heart sound and yields better time gates for heart sounds. PMID- 16311449 TI - The clinical application of non-invasive intracranial blood volume pulse wave monitoring. AB - An ultrasonic method was used to non-invasively measure intracranial blood volume (IBV) pulse waveforms. This technology has previously shown a strong association between invasively recorded ICP pulse waves and non-invasively recorded IBV pulse waves. The objective of the present study was to investigate the diagnostic value of non-invasively measured IBV pulse waves in the cases of different pathologies. A total of 75 patients were examined and these included cases of acute, chronic and stabilized hydrocephalus, spinal cord injury and terminal blood flow. These were compared to a control group of 53 healthy volunteers. The object of comparison was normalized and averaged IBV pulse waves. Pathological IBV pulse waveforms were compared with IBV pulse waveforms of the normal group using sub wave values, the area under waveform curve and the Euclidean distance calculation. The non-invasively measured IBV pulse waveform is not significantly dependent on acoustic path, gender or age. A detectable change in IBV pulse waveform shape was observed in situations when disturbance in intracranial hydrodynamics was present, e.g. during hypoventilation tests, in cases of terminal blood flow and hydrocephaly, depicting the level of hydrocephalus activity and the patient's compensatory capabilities as well as the effect of treatment. PMID- 16311450 TI - The influence of cardiac preload and positive end-expiratory pressure on the pre ejection period. AB - The pre-ejection period (PEP) has recently been described as a potential parameter for monitoring cardiac preload. This study further investigated the influence of changes in intravascular volume status and the application of positive end-expiratory pressure (PEEP) on the pre-ejection period. In ten pigs, ECG, arterial pressure and stroke volume derived from an aortic flowprobe were registered. Global end-diastolic volume (GEDV) was measured by transcardiopulmonary thermodilution. Total blood volume (TBV) and intrathoracic blood volume (ITBV) were measured by the dye-dilution technique. Measurements were performed during normovolaemic conditions, after volume loading with haemodilution blood (20 ml kg(-1)) and following haemorrhage (30 ml kg(-1)) without PEEP and with PEEP (15 cm H(2)O) applied. Volume loading increased GEDV, ITBV, TBV and SV, whereas PEP remained constant. However, the changes were not significant (P > 0.05). Subsequent haemorrhage significantly decreased GEDV (from 436 to 308 ml), ITBV (from 729 to 452 ml), TBV (from 2,131 to 1,488 ml) (all P values <0.05), and SV (from 20.7 ml to 14.3 ml, P < 0.001). However, PEP did not change significantly (from 73 to 82 ms, P > 0.05). No correlation between the changes in PEP and changes in any other variable was observed. It is concluded that PEP is not sensitive to the changes in intravascular volume status. PMID- 16311451 TI - Analysis of intracranial pressure during and after the infusion test in patients with communicating hydrocephalus. AB - The cerebrospinal fluid (CSF) infusion test is used to evaluate the dynamics of CSF circulation in patients with communicating hydrocephalus and is based on constant-rate infusion of the normal saline into cerebrospinal fluid space. The aim of the study was to refine methods of the analysis of intracranial pressure (ICP) recorded during and after the infusion test. The mathematical model of cerebrospinal fluid circulation was extended by the equation describing ICP decrease after the infusion. The nonlinear least-squares method of Levenberg Marquardt was used to estimate the parameters describing the CSF compensatory mechanisms. Twenty-seven infusion tests were studied. Both phases of the test-the increase and the decrease of ICP-were recorded and the compensatory parameters were calculated for each of them. ICP often does not return to the resting level after the infusion test within the period equivalent to the time of infusion in all cases. In 20 tests the differences between post- and pre-infusion resting ICP (DeltaICP) was higher than 1 mmHg, which was considered as significant. The mean value of DeltaICP for 20 infusion tests was 3.0 +/- 0.7 mmHg. The cerebral elasticity evaluated during the infusion was greater than the elasticity estimated from the decreasing phase after the infusion (0.24 +/- 0.07 ml(-1) versus 0.14 +/- 0.03 ml(-1); p < 0.01). PMID- 16311452 TI - Estimation of breathing interval from the photoplethysmographic signals in children. AB - Two important parameters that are generally under continual observation during clinical monitoring are heart rate (HR) variability and breathing interval (BI) of patients. Current HR monitoring during night-long childhood respiratory sleep studies is well tolerated but BI monitoring requires instrumentation, like nasal cannula, that can be less accommodating for children. In this study, BI was extracted from the photoplethysmographic (PPG) signals using a two-stage signal processing technique termed zero-phase digital filtering. Eight children (7 male) aged 8.6 +/- 2.6 years were recruited to perform two breathing activities: during tidal and with customized externally applied inspiratory resistive loading (IRL). The accuracy of BI derived from the PPG signals was compared with that estimated by a calibrated air pressure transducer in children. Statistical analysis revealed that mean BI attained from the PPG signals were significantly related during tidal breathing (r(2) = 0.76; range 0.61-0.83; p < 0.05) and with the IRL (r(2) = 0.79; range 0.68-0.85; p < 0.05) in the absence of motion artefacts. Preliminary findings herein suggest that besides having the capability to monitor HR and arterial blood oxygen saturation measurements, the PPG signals can be used to derive BI for children. This can be an attractive alternative for children who are more disturbed by intrusive techniques in prolonged clinical monitoring. PMID- 16311453 TI - Artifact reduction in magnetogastrography using fast independent component analysis. AB - The analysis of magnetogastrographic (MGG) signals has been limited to epochs of data with limited interference from extraneous signal components that are often present and may even dominate MGG data. Such artifacts can be of both biological (cardiac, intestinal and muscular activities, motion artifacts, etc) and non biological (environmental noise) origin. Conventional methods-such as Butterworth and Tchebyshev filters-can be of great use, but there are many disadvantages associated with them as well as with other typical filtering methods because a large amount of useful biological information can be lost, and there are many trade-offs between various filtering methods. Moreover, conventional filtering cannot always fully address the physicality of the signal-processing problem in terms of extracting specific signals due to particular biological sources of interest such as the stomach, heart and bowel. In this paper, we demonstrate the use of fast independent component analysis (FICA) for the removal of both biological and non-biological artifacts from multi-channel MGG recordings acquired using a superconducting quantum intereference device (SQUID) magnetometer. Specifically, we show that the signal of gastric electrical control activity (ECA) can be isolated from SQUID data as an independent component even in the presence of severe motion, cardiac and respiratory artifacts. The accuracy of the method is analyzed by comparing FICA-extracted versus electrode-measured respiratory signals. It is concluded that, with this method, reliable results may be obtained for a wide array of magnetic recording scenarios. PMID- 16311454 TI - Infrared ear thermometers--parameters influencing their reading and accuracy. AB - Infrared ear thermometers (IRETs) are extensively used for measuring the temperature of a human body. For accurate measurements with IRETs they have to be calibrated regularly with an appropriate and traceable calibration system. Such systems are neither widely available nor are there many competent (accredited) laboratories which can provide traceability for IRETs. This paper describes some important influential parameters in the calibration and use of IRETs, which were observed during the extensive research on IRETs and have not been reported in the literature yet. IRET readings, and consequently also their most important metrological characteristics, accuracy and uncertainty of measurement, depend on these influential parameters. According to our findings, we would like to warn users of medical radiation thermometers, not only IRETs but also forehead thermometers and infrared temperature scanning systems, that they should be extremely careful in selection, maintenance and use of medical radiation thermometers. Measurement accuracy, as it is required by several technical standards, is hard to achieve with the majority of currently available medical radiation thermometers. PMID- 16311455 TI - Clinical comparison of tympanic membrane displacement with invasive intracranial pressure measurements. AB - A non-invasive method of assessing intracranial pressure (ICP) would be of benefit to patients with abnormal cerebral pathology that could give rise to changes in ICP. In particular, it would assist the regular monitoring of hydrocephalus patients. This study evaluated a technique using tympanic membrane displacement (TMD) measurements, which has been reported to provide a reliable, non-invasive measure of ICP. A group of 135 hydrocephalus patients was studied, as well as 13 patients with benign intracranial hypertension and a control group of 77 volunteers. TMD measurements were carried out using the Marchbanks measurement system analyser and compared between the groups. In 36 patients, invasive measurements of ICP carried out at the same time were compared with the TMD values. A highly significant relationship was found between TMD and ICP but intersubject variability was high and the predictive value of the technique low. Taking the normal range of ICP to be 10-15 mmHg, the predictive limits of the regression are an order of magnitude wider than this and therefore Vm cannot be used as a surrogate for ICP. In conclusion, TMD measurements do not provide a reliable non-invasive measure of ICP in patients with shunted hydrocephalus. PMID- 16311456 TI - Pulse transit time changes observed with different limb positions. AB - Pulse transit time (PTT) is a non-invasive measure of arterial compliance. It can be used to assess instantaneous blood pressure (BP) changes in continual cardiovascular measurement such as during overnight respiratory sleep studies. In these studies, periodic changes in limb position can occur randomly. However, little is known about their possible effects on PTT monitored on the various limbs. The objective of this study was to evaluate PTT differences on all four limbs during two positional changes (lowering and raising of a limb). Ten healthy adults (seven male) with a mean age of 27.0 years were recruited in this study. The results showed that the limb that underwent a positional change had significant (p < 0.05) local PTT differences when compared to its nominal baseline value, whereas PTT changes in the other remaining limbs were insignificant (p > 0.05). The mean PTT value measured from a vertically-raised limb increased by 42.7 ms, while it decreased by 28.1 ms with a half-lowered limb. The PTT differences observed during positional change can be contributed to by the complex interactions between hydrostatic pressure changes, autonomic and local autoregulation experienced in these limbs. Hence the findings herein suggest that PTT is able to reflect local circulatory responses despite changes in the position of other limbs. This can be useful in prolonged clinical observations where limb movements are expected. PMID- 16311457 TI - Effects of lung time constant, gas analyser delay and rise time on measurements of respiratory dead-space. AB - This study evaluated effects of mechanical time constants (tau(m)) of the respiratory system, delays between flow and CO(2) partial pressure (P(CO)(2)) signals and rise time of the CO(2) analyser on dead-space measurements. A computer model simulated low alveolar dead-space, high alveolar dead-space, 0.2 80%) of the power of noninvasive signals was linearly related to the power of the invasive signal. The characteristic frequency of each band was substantially preserved in both noninvasive signals. The results for diastolic pressure were similar, but the Finapres errors in the VLF and LF bands were lower. Baroreflex sensitivity was significantly underestimated by both devices (Finometer: -31%, Finapres: -24%). Despite previous studies having shown that brachial artery waveform reconstruction performed by the Finometer has improved the accuracy of blood pressure measurement compared to the Finapres device, measurement of blood pressure variability in cardiac disease patients provides worse results in most spectral parameters and a better accuracy only in the HF band of systolic pressure. PMID- 16311460 TI - Assessment of cerebrospinal fluid outflow conductance using constant-pressure infusion--a method with real time estimation of reliability. AB - The outflow conductance (C(out)) of the cerebrospinal fluid (CSF) system is a parameter considered to be predictive in selection for hydrocephalus surgery. C(out) can be determined through an infusion test. A new apparatus for performing infusion tests in a standardized and automated way was developed. The objective was to evaluate repetitiveness as well as to propose and evaluate a method for real time estimation of the reliability of individual C(out) investigations. Repeated investigations were performed on an experimental model simulating the CSF system, and on 14 patients with hydrocephalus. DeltaC(out), calculated as the 95% confidence interval of C(out), was introduced as an estimate of the reliability of individual C(out) investigations. On the model, no significant difference was found between DeltaC(out) and the actual C(out) variation in repeated investigations (p = 0.135). The correlation between the first and the second patient investigation was high (R = 0.99, p < 0.05), although there was a significant difference between the investigations (p < 0.05). The standard deviation of difference was 2.60 microl (s kPa)(-1). The repetitiveness of C(out) with the new apparatus was high, and DeltaC(out) reflected the reliability of each investigation. This feature has to be taken into account in every individual case, before making a decision or performing research based on measurements of C(out) in the future. PMID- 16311485 TI - Social justice and community health centers: commitment to one gave rise to the other. PMID- 16311486 TI - Stymied. PMID- 16311487 TI - Homeless, mentally ill and addicted: the need for abuse and trauma services. AB - This paper examines an empirical investigation of the lifetime prevalence of trauma (defined as sexual and/or physical abuse) in a cohort of adults enrolled in a federally funded initiative that provides treatment for homeless persons suffering the effects of comorbid substance use and serious mental illness, and considers the impact of this information on clinical programming. Data collected from homeless individuals with co-occurring disorders admitted to the Seeking Treatment and Recovery (STAR) Program during a one year period (n=78) were analyzed for a history of trauma events. Of those individuals evaluated, 79.5% (62/78) acknowledged a history of either physical and/or sexual abuse at some time in their lifetimes. Of this population, 100% of the homeless women (27/27) with co-occurring disorders had experienced a life-altering traumatic event while 68.6% (35/51) of the homeless men also reported trauma histories. We describe the trauma-based interventions made in the STAR Program that have the potential for replication in other initiatives committed to serving homeless individuals with co-occurring disorders. PMID- 16311488 TI - Universal health care coverage for children: impact on pediatric health care providers. AB - A Northern California county expanded health coverage to cover nearly all children in the state through a new insurance program. In two years, 75,500 children entered a health care system near capacity. We hypothesized that the influx of thousands of previously uninsured children into the health system would affect providers in many ways. This cross-sectional study sought to investigate how this influx affected provider practices, job satisfaction, access to specialists, and overarching views about the program. Qualitative analyses of expert interviews were performed. Providers reported improved access to health care, specialists, and medications for patients. They cited increased job satisfaction for providers due to fewer limits on care, improved referral process, and decreased patient family financial stress. Providers noted the persistence of long appointment wait times for specialist care. After moving to near universal coverage, safety net providers described increased job satisfaction. Because this study examined safety-net providers, future research requires a more representative sample of providers. PMID- 16311489 TI - Cancer screening among Native Americans in eastern North Carolina. AB - The purpose of the study is to explore cancer screening predictors and practices among Lumbee Indians in eastern North Carolina. Interviewers from the community conducted phone interviews with 333 men and 456 women aged 40 and older randomly selected from the Lumbee Tribal roll as part of the Lumbee Diabetes and Health Survey. The survey has sections on demographic information and health conditions, family history of chronic conditions, and health behaviors. This paper reports on cancer screening behaviors. Rates of ever being screened for breast, cervical, prostate and colon cancer were relatively high compared with overall national rates. Predictors included younger age, better health, more education, and lifestyle factors such as engaging in regular physical activity and not smoking. Future research should focus on developing culturally appropriate campaigns to increase the frequency of cancer screening to conform to guidelines, and educational programs and interventions to reach Lumbee Indians most at risk for not being screened. PMID- 16311490 TI - Health insurance coverage and health-related quality of life: analysis of 2000 Medical Expenditure Panel Survey data. AB - This study investigated relationships between health insurance status and health related quality of life (HRQOL) using the 2000 Medical Expenditure Panel Survey data. Health-related quality of life was measured using the SF-12 Physical Component Summary (PCS) and SF-12 Mental Component Summary (MCS). The analysis controlled for sociodemographic and attitudinal variables and medical conditions. The analysis also investigated and controlled for possible reverse causality between HRQOL and health insurance in the models. After adjusting for covariates, individuals without health insurance had significantly lower mean PCS scores (beta=-5.8; SE=0.4) than those with health insurance. The adjusted association between no health insurance and MCS scores (beta=-1.1; SE=0.4) also was significant. The adjusted difference in HRQOL among people with health insurance and those without it exceeds or is comparable to adjusted differences in HRQOL between people with each of various medical conditions and people without them. PMID- 16311491 TI - The effect of socioeconomic status on the survival of people receiving care for HIV infection in the United States. AB - HIV-infected people with low socioeconomic status (SES) and people who are members of a racial or ethnic minority have been found to receive fewer services, including treatment with Highly Active Antiretroviral Therapy (HAART), than others. We examined whether these groups also have worse survival than others and the degree to which service use and antiretroviral medications explain these disparities in a prospective cohort study of a national probability sample of 2,864 adults receiving HIV care. The independent variables were wealth (net accumulated financial assets), annual income, educational attainment, employment status (currently working or not working), race/ethnicity, insurance status, use of services, and use of medications at baseline. The main outcome variable was death between January 1996 and December 2000. The analysis was descriptive and multivariate adjusted Cox proportional hazards regression analysis of survival. By December 2000, 20% (13% from HIV, 7% non-HIV causes) of the sample had died. Those with no accumulated financial assets had an 89% greater risk of death (RR=1.89, 95% CI=1.15-3.13) and those with less than a high school education had a 53% greater risk of death (RR=1.53, 95% CI=1.15-2.04 ) than their counterparts, after adjusting for sociodemographic and clinical variables only. Further adjusting for use of services and antiretroviral treatment diminished, but did not eliminate, the elevated relative risk of death for those with low SES by three of the four measures. The finding of markedly elevated relative risks of death for those with HIV infection and low SES is of particular concern given the disproportionate rates of HIV infection in these groups. Effective interventions are needed to improve outcomes for low SES groups with HIV infection. PMID- 16311492 TI - Optimal design features for surveying low-income populations. AB - Improving Medicaid program effectiveness for underserved populations is hampered by low survey response rates. This study determined how to maximize Medicaid consumer satisfaction survey response rates to the Consumer Assessment of Health Plans Study (CAHPS) survey. In a public immunization clinic, 8 focus groups and 15 extended interviews were used to assess consumer-preferred survey design features and incentives. To test hypotheses, we conducted the following trial. Out of 10,733 total participants in a Kansas Medicaid managed care plan, 3,685 eligible for CAHPS were unduplicated by household. After randomization of the 968 households with valid addresses to one of three groups, a controlled trial was conducted to assess response rates to CAHPS survey formats and incentives. Response rates were 35% for a standard mailing, 44% for a user-friendly low literacy mailing, and 64% for a user-friendly low-literacy mailing with a $10 contingent incentive. Both experimental arms significantly improved response compared with the control; the response rate of the mailing group with the incentive was higher than the response rate of the group receiving that mailing without any incentive (p<0.0001). Using consumer-based preferences significantly increased response rates to this Medicaid satisfaction survey. Raising CAHPS response rates may increase validity of Medicaid consumer satisfaction information. PMID- 16311493 TI - Identifying areas for improvement: results of a Medicaid managed care diabetes survey. AB - This study explores factors that contribute to dissatisfaction for Medicaid managed care enrollees with diabetes. Using results from a mail survey of 2,104 Medicaid managed care enrollees with diabetes, multivariate logistic regression models were fit for 3 outcomes: dissatisfaction with diabetes information from doctor or nurse, dissatisfaction with diabetes care received and dissatisfaction with health plan. Across the 3 models, enrollees who reported they were in poor health, who failed to engage in self-management activities, who reported little or inadequate patient education and/or who had difficulty accessing diabetes care were more likely to report dissatisfaction. The results of this study will be used to propose interventions that Medicaid managed care plans can initiate to improve satisfaction of enrollees with diabetes. PMID- 16311494 TI - Predictors of improved HbA1c testing by primary care physicians. AB - Measurement of glycosylated hemoglobin (HbA1c) among patients with diabetes mellitus contributes to attaining control of blood glucose, which in turn is associated with fewer complications. Here, we seek to identify physician and county level characteristics that predict increased HbA1c testing among Medicare beneficiaries with diabetes who did not have testing in the baseline year. Physicians in counties with more people on Medicaid, or with fewer physicians per capita tended to have less improvement in HbA1c testing among Medicare beneficiaries over the three year study period. PMID- 16311495 TI - Limited health care access impairs glycemic control in low income urban African Americans with type 2 diabetes. AB - Limited access to health care is associated with adverse outcomes, but few studies have examined its effect on glycemic control in minority populations. Our observational cross-sectional study examined whether differences in health care access affected hemoglobin A1c (HbA1c) levels in 605 patients with diabetes (56% women; 89% African American; average age, 50 years; 95% with type 2 diabetes) initially treated at a municipal diabetes clinic. Patients who had difficulty obtaining care had higher A1c levels (9.4% vs. 8.7%; p=0.001), as did patients who used acute care facilities (9.5%; p<0.001) or who had no usual source of care (10.3%; p<0.001) compared with those who sought care at doctors' offices or clinics (8.6%). In adjusted analyses, HbA1c was higher in persons who gave a history of trouble obtaining medical care (0.57%; p=0.04), among persons who primarily used an acute care facility to receive their health care (0.49%; p=0.047), and in patients who reported not having a usual source of care (1.08%; p=0.009). Policy decisions for improving diabetes outcomes should target barriers to health care access and focus on developing programs to help high-risk populations maintain a regular place of health care. PMID- 16311496 TI - Community's voice in managing childhood asthma: the assessment phase of research. AB - This study was designed to investigate community beliefs about caring for childhood asthma and to elicit suggestions for interventions to improve asthmatic children's health. Focus groups were conducted with parents of children with asthma, children with asthma, school staff, and health care and childcare professionals. Data were analyzed for themes, such as disruption of normal living and having to work in a chaotic system, enabling researchers to posit a core belief for each group. These core beliefs, together with encompassed other, related beliefs held by group members, guide attitudes and actions about asthma. Interventions recommended by focus group participants included creating an asthma play, asthma education, and developing a clinic-based registry to standardize asthma documentation. The community's voice is important in assessment and design of health improvement projects. Incorporating the community's suggestions gives the community a sense of contributing to the health care of their children with asthma. PMID- 16311497 TI - Medicaid enrollee switching among managed care plans. AB - This case-study (n=41,969) of voluntary switching among Mecklenburg County, North Carolina Medicaid managed care plans showed a low switching rate of 14.5 per 100 member-years over 33 months, or 5.3 averaged annually. Population, plan and plan characteristics, claims and telephone survey data were examined to better understand this important behavioral measure. Switching in Medicaid managed care, which is little studied, is contrasted with the extensive literature on middle class switching. Policy implications included the suitability of Medicaid populations for managed care and the need for more research on switching and disenrollment and the Medicaid innovation of neutral health benefits advising. PMID- 16311498 TI - Detection of depression among low-income Mexican Americans in primary care. AB - Between one and two thirds of depressive disorders go undetected in primary care settings. Four hundred ninety-six Mexican American primary care patients from high-poverty areas were screened for depressive symptoms, and 41% endorsed depressive symptoms. Eighty percent of screened patients with depressive symptoms agreed to structured diagnostic interviews and 90% of those interviewed met diagnostic criteria for one or more depressive disorders. Cases of depression detected through this systematic process were compared with evidence of depression detected by providers in medical charts. Provider and study evaluation agreement was poor (kappa= 0.13); providers noted depression in 21% of patients with depressive disorders based on the systematic evaluation. More work is needed to enhance detection of depression in primary care, especially in minority populations. PMID- 16311499 TI - Managing anemia in low-income toddlers: barriers, challenges and context in primary care. AB - Iron-deficiency remains a concern among low-income toddlers in the U.S. This formative study describes how primary care providers serving high-risk 1- to 3 year-old children in an urban ambulatory care setting approach anemia. Data collection included a retrospective review of randomly selected medical records (n=264) and semi-structured interviews with clinicians (n=41). Thirty-eight percent of the children presented with anemia (Hgb < 11.0 g/dl) at least once between 12 and 36 months of age. Just under half of these children were treated for anemia. Follow-up laboratories for iron-treated children were completed within 35 days in 16% of cases (median: 3 months). Interviews identified four key themes (iron-deficiency, communication, poverty, system) running through the two major categories of prevention and treatment. Treatment cut-points were variable. While providers felt clinically comfortable with anemia, they felt burdened and challenged by follow-up. Communication and system barriers weighed most heavily on perceived treatment outcomes. PMID- 16311500 TI - Provider behaviors contributing to patient self-management of chronic illness among underserved populations. AB - Effective treatment of chronic illness requires active engagement of patients in managing their own conditions. This research explores the influence of provider support for self-management on patients' performance of self-care activities, a widely presumed but under-studied relationship, particularly among underserved populations. We surveyed an ethnically diverse, primarily low-income sample of 956 patients (or parents of pediatric patients) with diabetes or asthma in 17 outpatient teaching settings across the country. Multivariate analysis established that patients with strongly positive assessments of their providers were substantially more confident in self-care. Further, among patients with diabetes, high assessments of provider support, in comparison with low, were significantly associated with performance of self-management tasks, amounting to approximately one more day per week. These relationships were significant for patients with either illness and from varied socio-economic backgrounds. The results provide evidence of the validity of our measure of provider support, its relevance to underserved populations, and its usefulness for evaluating quality of care. PMID- 16311503 TI - Bilateral choanal atresia in premature monozygotic twins. AB - Bilateral congenital choanal atresia is a relatively rare anomaly of the upper airway, which may cause life-threatening respiratory emergency and require rapid diagnosis and treatment. This condition usually occurs sporadically, but has also been rarely described in siblings. We present monozygotic premature twin infants with identical findings of bilateral choanal atresia and no other associated anomalies. To our knowledge, this is the first report of such an occurrence. PMID- 16311504 TI - Management of severe hyperbilirubinemia in the newborn: adrenal hematoma revisited. AB - A 4-day-old male infant presented with complaints of jaundice on the third day of life. He was full-term and appropriate for gestational age and born to unrelated parents. All laboratory investigation tests were normal except total serum bilirubin of 27.4 mg/dl with a direct bilirubin 0.29 mg/dl. Abdominal and cranial ultrasonography (US) was performed on sixth day of life because of severe hyperbilirubinemia. Abdominal US revealed adrenal hematoma. Enclosed hematomas may cause significant unconjugated hyperbilirubinemia in absence of other high risk conditions. PMID- 16311505 TI - Prophylactic Phototherapy in ABO Incompatibility. PMID- 16311507 TI - The PML-nuclear body associated protein Daxx regulates the cellular response to CD40. PMID- 16311508 TI - Involvement of glutaredoxin-1 and thioredoxin-1 in beta-amyloid toxicity and Alzheimer's disease. AB - Strong evidence indicates oxidative stress in the pathogenesis of Alzheimer's disease (AD). Amyloid beta (Abeta) has been implicated in both oxidative stress mechanisms and in neuronal apoptosis. Glutaredoxin-1 (GRX1) and thioredoxin-1 (TRX1) are antioxidants that can inhibit apoptosis signal-regulating kinase (ASK1). We examined levels of GRX1 and TRX1 in AD brain as well as their effects on Abeta neurotoxicity. We show an increase in GRX1 and a decrease in neuronal TRX1 in AD brains. Using SH-SY5Y cells, we demonstrate that Abeta causes an oxidation of both GRX1 and TRX1, and nuclear export of Daxx, a protein downstream of ASK1. Abeta toxicity was inhibited by insulin-like growth factor-I (IGF-I) and by overexpressing GRX1 or TRX1. Thus, Abeta neurotoxicity might be mediated by oxidation of GRX1 or TRX1 and subsequent activation of the ASK1 cascade. Deregulation of GRX1 and TRX1 antioxidant systems could be important events in AD pathogenesis. PMID- 16311509 TI - Pharmacological induction of Hsp70 protects apoptosis-prone cells from doxorubicin: comparison with caspase-inhibitor- and cycle-arrest-mediated cytoprotection. AB - Selective modulation of cell death is important for rational chemotherapy. By depleting Hsp90-client oncoproteins, geldanamycin (GA) and 17-allylamino-17 demethoxy-GA (17-AAG) (heat-shock protein-90-active drugs) render certain oncoprotein-addictive cancer cells sensitive to chemotherapy. Here we investigated effects of GA and 17-AAG in apoptosis-prone cells such as HL60 and U937. In these cells, doxorubicin (DOX) caused rapid apoptosis, whereas GA induced heat-shock protein-70 (Hsp70) (a potent inhibitor of apoptosis) and G1 arrest without significant apoptosis. GA blocked caspase activation and apoptosis and delayed cell death caused by DOX. Inhibitors of translation and transcription and siRNA Hsp70 abrogated cytoprotective effects of GA. Also GA failed to protect HL60 cells from cytotoxicity of actinomycin D and flavopiridol (FL), inhibitors of transcription. We next compared cytoprotection by GA-induced Hsp70, caspase inhibitors (Z-VAD-fmk) and cell-cycle arrest. Whereas cell-cycle arrest protected HL60 cells from paclitaxel (PTX) but not from FL and DOX, Z-VAD-fmk prevented FL induced apoptosis but was less effective against DOX and PTX. Thus, by inducing Hsp70, GA protected apoptosis-prone cells in unique and cell-type selective manner. Since GA does not protect apoptosis-reluctant cancer cells, we envision a therapeutic strategy to decrease side effects of chemotherapy without affecting its therapeutic efficacy. PMID- 16311511 TI - Role of glucose-6-phosphate dehydrogenase for oxidative stress and apoptosis. PMID- 16311512 TI - Regulation of the antioncogenic Chk2 kinase by the oncogenic Wip1 phosphatase. AB - The antioncogenic Chk2 kinase plays a crucial role in DNA damage-induced cell cycle checkpoint regulation. Here we show that Chk2 associates with the oncogenic protein Wip1 (wild-type p53-inducible phosphatase 1) (PPM1D), a p53-inducible protein phosphatase. Phosphorylation of Chk2 at threonine68 (Thr68), a critical event for Chk2 activation, which is normally induced by DNA damage or overexpression of Chk2, is inhibited by expression of wild-type (WT), but not a phosphatase-deficient mutant (D314A) of Wip1 in cultured cells. Furthermore, an in vitro phosphatase assay revealed that Wip1 (WT), but not Wip1 (D314A), dephosphorylates Thr68 on phosphorylated Chk2 in vitro, resulting in the inhibition of Chk2 kinase activity toward glutathione S-transferase-Cdc25C. Moreover, inhibition of Wip1 expression by RNA interference results in abnormally sustained Thr68 phosphorylation of Chk2 and increased susceptibility of cells in response to DNA damage, indicating that Wip1 acts as a negative regulator of Chk2 in response to DNA damage. PMID- 16311513 TI - BMP-4 induces a Smad-dependent apoptotic cell death of mouse embryonic stem cell derived neural precursors. AB - Embryonic ectoderm is fated to become either neural or epidermal, depending on patterning processes that occur before and during gastrulation. It has been stated that epidermal commitment proceeds from a bone morphogenetic protein-4 (BMP-4)-dependent inhibition of dorsal ectoderm neuralization. We recently demonstrated that murine embryonic stem (ES) cells treated with BMP-4 undergo effective keratinocyte commitment and epidermogenesis. Focusing on the precise role of BMP-4 in the early choice between neural and epidermal commitment, we show here that BMP-4 treatment of ES cells leads to a dramatic apoptotic death of Sox-1+ neural precursors with concomitant epidermal engagement. In addition, neutralization of the Smad pathway prevents both the BMP-4 apoptotic process and the inhibition of neural differentiation. Our results suggest that, in mammals, BMP-4, as an active inducer of epidermal commitment, interferes with the survival of neural precursors through induction of their apoptotic cell death. PMID- 16311514 TI - CaM-kinaseII-dependent commitment to microcystin-induced apoptosis is coupled to cell budding, but not to shrinkage or chromatin hypercondensation. AB - The protein phosphatase inhibitor microcystin-LR (MC) induced hepatocyte apoptosis mediated by the calcium-calmodulin-dependent multifunctional protein kinase II (CaMKII). CaMKII antagonists were added at various times after MC to define for how long the cells depended on CaMKII activity to be committed to execute the various parameters of death. Shrinkage and nonpolarized budding were reversible and not coupled to commitment. A critical commitment step was observed 15-20 min after MC (0.5 microM) addition. After this, CaMKII inhibitors no longer protected against polarized budding, DNA fragmentation, lost protein synthesis capability, and cell disruption. Commitment to chromatin hypercondensation occurred 40 min after MC addition. In conclusion, irreversible death commitment was coupled to polarized budding, but not to shrinkage or chromatin condensation. Antioxidant prevented chromatin condensation when given after the CaMKII dependent commitment point, suggesting that CaMKII had mediated the accumulation of a second messenger of reactive oxygen species nature. PMID- 16311515 TI - Apoptosis and differentiation commitment: novel insights revealed by gene profiling studies in mouse embryonic stem cells. AB - Mouse embryonic stem (ES) cells remain pluripotent in vitro when grown in the presence of leukemia inhibitory factor (LIF). LIF starvation leads to apoptosis of some of the ES-derived differentiated cells, together with p38alpha mitogen activated protein kinase (MAPK) activation. Apoptosis, but not morphological cell differentiation, is blocked by a p38 inhibitor, PD169316. To further understand the mechanism of action of this compound, we have identified its specific targets by microarray studies. We report on the global expression profiles of genes expressed at 3 days upon LIF withdrawal (d3) compared to pluripotent cells and of genes whose expression is modulated at d3 under anti-apoptotic conditions. We showed that at d3 without LIF cells express, earlier than anticipated, specialized cell markers and that when the apoptotic process was impaired, expression of differentiation markers was altered. In addition, functional tests revealed properties of anti-apoptotic proteins not to alter cell pluripotency and a novel role for metallothionein 1 gene, which prevents apoptosis of early differentiated cells. PMID- 16311516 TI - The KSHV oncoprotein vFLIP contains a TRAF-interacting motif and requires TRAF2 and TRAF3 for signalling. AB - Primary effusion lymphomas (PELs) characterized by infection with the Kaposi's sarcoma herpesvirus (KSHV; also called human herpesvirus 8) depend on the expression of the viral FADD-like interleukin-1-beta-converting enzyme (FLICE)/caspase-8-inhibitory protein (vFLIP) for their survival. This effect is achieved by activation of the transcription factor nuclear factor-kappaB (NF kappaB). Tumour necrosis factor (TNF) receptor-associated factors (TRAFs) are direct mediators of NF-kappaB signalling by TNF family receptors and the Epstein Barr virus oncoprotein latent membrane protein 1 and so we assessed the role of TRAFs in signalling by vFLIP. Here, we report the identification of a TRAF interacting motif (PYQLT) in vFLIP, which is not present in other FLIP molecules. We show that vFLIP directly binds to TRAF2 in vitro and in PEL cells. TRAF2 and TRAF3 are required for induction of NF-kappaB and associated cell survival, as well as Jun amino-terminal kinase phosphorylation by vFLIP, whereas TRAF1, TRAF5 and TRAF6 are dispensable. Mutations in the P93 or Q95 amino acids within the TRAF-interacting motif of vFLIP abolish its ability to bind to TRAF2 and to signal to NF-kappaB. TRAF2, but not TRAF3, mediates the association of vFLIP with the IkappaB kinase complex. These data indicate that vFLIP uses TRAF2 and TRAF3 for signalling to NF-kappaB, which is crucial for KSHV-associated lymphomagenesis. PMID- 16311517 TI - The role of AtMSH2 in homologous recombination in Arabidopsis thaliana. AB - During homologous recombination (HR), a heteroduplex DNA is formed as a consequence of strand invasion. When the two homologous strands differ in sequence, a mismatch is generated. Earlier studies showed that mismatched heteroduplex often triggers abortion of recombination and that a pivotal component of this pathway is the mismatch repair Msh2 protein. In this study, we analysed the roles of AtMSH2 in suppression of recombination in Arabidopsis. We report that AtMSH2 has a broad range of anti-recombination effects: it suppresses recombination between divergent direct repeats in somatic cells or between homologues from different ecotypes during meiosis. This is the first example of a plant gene that affects HR as a function of sequence divergence and that has an anti-recombination meiotic effect. We discuss the implications of these results for plant improvement by gene transfer across species. PMID- 16311518 TI - Repression of polymerase I-mediated gene expression at Trypanosoma brucei telomeres. AB - The African trypanosome, Trypanosoma brucei, is a flagellated pathogenic protozoan that branched early from the eukaryotic lineage. Unusually, it uses RNA polymerase I (Pol I) for mono-telomeric expression of variant surface glycoprotein (VSG) genes in bloodstream-form cells. Many other subtelomeric VSG genes are reversibly repressed, but no repressive DNA sequence has been identified in any trypanosomatid. Here, we show that artificially seeded de novo telomeres repress Pol I-dependent gene expression in mammalian bloodstream and insect life-cycle stages of T. brucei. In a telomeric VSG expression site, repression spreads further along the chromosome and this effect is specific to the bloodstream stage. We also show that de novo telomere extension is telomerase dependent and that the rate of extension correlates with the expression level of the adjacent gene. Our results show constitutive telomeric repression in T. brucei and indicate that an enhanced, developmental stage-specific repression mechanism controls antigenic variation. PMID- 16311519 TI - Control of myelination in Schwann cells: a Krox20 cis-regulatory element integrates Oct6, Brn2 and Sox10 activities. AB - Myelination in Schwann cells is governed by several transcription factors, including the POU proteins Oct6 and Brn2, the high mobility group protein Sox10 and the zinc-finger protein Krox20. How the function of these factors is integrated in the control of myelination has not been established. Previously, we identified an enhancer element controlling Krox20 expression throughout myelination in Schwann cells. In this paper, cell culture experiments were combined with transgenesis to identify transcription factors acting directly upstream of Krox20. The results show that during the promyelin-myelin transition, Krox20 expression is directly activated by Oct6 and Brn2 acting on this enhancer. In addition, the enhancer-dependent synergism between these POU proteins and Sox10 suggests that Krox20 expression requires this combination of factors. These results resolve previous controversy concerning the mechanism of action of Oct6 and Brn2 during myelination and provide an explanation for myelin deficiencies in Waardenberg-Hirschsprung disease patients whereby Sox10 mutations could lead to a loss of Krox20 expression. PMID- 16311520 TI - Massive haemorrhagic retinal detachment after transpupillary thermotherapy for choroidal neovascularisation. PMID- 16311521 TI - Glaucoma management in pregnancy: a questionnaire survey. AB - BACKGROUND: The management of glaucoma in a pregnant woman can pose a challenge for the ophthalmologist. The risks and benefits to the mother and foetus must be weighed. When it is decided that treatment is necessary, there is no consensus as to the safest treatment. This study was designed to garner the experiences and views of UK ophthalmologists with regard to this scenario. METHODS: All consultant ophthalmologists in the UK were sent a questionnaire. They were asked if they had previous experience treating pregnant women with glaucoma, what management they had used, what management they would currently employ, and, if they were to use medical treatment, what would their first choice agent be. RESULTS: A total of 282 out of 605 questionnaires were returned. Of the respondents, 26% had previously treated pregnant women with glaucoma. Most (71%) had continued with the therapy that the women were already on. In all, 34% had observed the situation and had not needed to treat. When asked what they would currently do in this scenario, 31% were unsure. A total of 40% would simply continue the prepregnancy treatment. A total of 45% of respondents, if they were to use medical treatment, would use beta-blockers, compared to 33% who would use prostaglandins. CONCLUSION: Although it is thought to be a relatively rare clinical situation, over a quarter of our respondents had first hand experience of the management of glaucoma in pregnancy. Our results indicate some uncertainty about the optimal strategy. It is unlikely that trials will be performed and we suggest guidelines be produced using current knowledge and modified as evidence appears. PMID- 16311522 TI - Deep sclerectomy with mitomycin C in failed trabeculectomy. AB - AIM: To evaluate the success rate and complications associated with deep sclerectomy with mitomycin C (MMC) and a reticulated hyaluronic acid implant in previously failed trabeculectomy. METHODS: This prospective study included 20 eyes with a previously failed trabeculectomy, which were treated with deep sclerectomy with 0.2 mg/ml MMC application under the conjunctiva and superficial scleral flap. Intraocular pressure (IOP), glaucoma medications, visual acuity, and complications were recorded preoperatively and 1 day, 1 week, and 1, 3, 6, and 12 months postoperatively. RESULTS: The mean preoperative IOP was 25.8+/-7.3 mmHg; the IOP significantly decreased to 14.6+/-3.2 mmHg 1 year postoperative. At each interval, the mean IOP was significantly lower than preoperatively (P=0.000). At 1 year, the complete success rate (IOP0.05). However, the proportion of the distance between the porta hepatis and the upper margin to the distance between the porta hepatis and the lower margin decreased significantly with gestational age (p<0.05). Type 3 liver (square) was the most commonly observed type of fetal liver (53%). CONCLUSION: Our opinion is that the parameters obtained can be useful to diagnose pathologies of liver development and anomalies concerning several branches of medicine such as anatomy, pathologic anatomy (fetopathology), forensic medicine, medical imaging, obstetrics and pediatrics. PMID- 16311654 TI - Cell division and cellular morphology of the chick retinal pigmented epithelial cells in culture: a time-lapse analysis. AB - OBJECTIVE: To investigate the patterns of cell division, movement and shape during early stages of development of the chick embryo retinal pigmented epithelial (RPE) cells and to evaluate the morphology of dissociated embryonic cells with regard to their proliferation capacity. METHODS: We conducted this study at the Department of Histology and Embryology, Celal Bayar University, Manisa, Turkey, between 2002 and 2003. We isolated the cells from chick embryos. We analyzed the images of the embryonic cells originated from neuroepithelia using a computer-based time-lapse acquisition system attached to a differential interference contrast microscope. RESULTS: Retinal pigmented epithelial cells, despite being dissociated, depict a colony-type growth. Cells in the periphery of the colony and those outside the colony showed a tendency to proliferate and migrate and retained contact with the neighboring cells during division. Characteristics of cytokinesis were separation from the neighboring cell while retaining an attachment point, became rounded, moved up and started to shake and ascend to disseminate to the substrate to complete the division. The round-up stage was non-significantly shorter when the cell was closer to the center of the colony. Cells that were in the periphery of, or outside the colony had a round-up time of over one hour while cytokinesis-to-adhesion time was around 5 minutes. However, when we found the cells in the center of the colony, the times were half an-hour and 1.5 hours for the daughter cells, a 2-fold difference between daughter cells with regard to the duration of attachment. CONCLUSION: Cell division, migration and proliferation are complex procedures influenced by growth factors, cell adhesion, matrix molecules underneath and the signal mechanisms and can be studied in detail using time-lapse microscopy, immunohistochemistry and confocal microscopy. PMID- 16311655 TI - Construction of intrauterine growth curves in a high altitude area of Saudi Arabia. AB - OBJECTIVE: The aim of the present study was to construct intrauterine percentile growth curves for body weight, length and head circumference for local use in a high altitude area of Saudi Arabia. METHODS: This is a cross-sectional study of all Saudi births from Abha General Hospital over a 6-year period from 1999 to 2004. We included a total of 6,035 Saudi births in the present study after eliminating babies that were stillbirths, twins or those with major congenital anomalies. The gestational age of the infants ranged from 26-42 weeks. The anthropometric measurements included birth weight, crown-heel length and head circumference. We calculated the 10th, 25th, 50th, 75th, and 90th percentiles for weight, length and head circumference against the periods of gestation. We also calculated the mean weight and mean +/- 2SD. We determined the curves of best fit for weight, length and head circumference measurements at different weeks of gestation by polynomial regression of the following general form to construct the clinical curves: Y=a + bX + cX2. RESULTS: We derived intrauterine growth curves for weight, length and head circumference from measurements made on infants born at each week of gestation in the latter half of pregnancy. By calculating mean values and deviations around these (expressed either as centiles or standard deviations), we constructed distance (size attained) growth curves. The mean values for weight and length of births of the present study are lower than those of published charts in all gestational periods. However, for head circumference, evident appeared only before 32 weeks of gestation. The mean values of head circumference were nearly comparable. CONCLUSION: Constructed smoothed gestational curves are a useful tool for assessing the intrauterine growth of births in high altitude areas of Saudi Arabia. PMID- 16311656 TI - Lung function in type 1 Saudi diabetic patients. AB - OBJECTIVE: To determine the effects of type 1 diabetes mellitus on lung function and its gravity in relation to the duration of disease. METHODS: We carried out this study in the Department of Physiology, College of Medicine and Diabetic Center, King Abdul-Aziz University Hospital, King Saud University, Riyadh, Saudi Arabia during the period 2003-2004. We randomly selected a group of 27 apparently healthy volunteer Saudi male type 1 diabetic patients with age ranging from 19-70 years. We matched the diabetic patients with another group of 27 control healthy male subjects in terms of age, height, weight and socioeconomic status. Both groups met with exclusion criteria as per standard. We performed spirometry on an Electronic Spirometer (Schiller AT-2 Plus, Switzerland) and compared the results by a student t-test (2-tailed). RESULTS: Type 1 diabetic patients showed a significant reduction in the forced vital capacity and forced expiratory volume in one second (FEV1) relative to their matched controls. However, there was no significant difference in the forced expiratory ratio, forced expiratory flow; forced expiratory flow and peak expiratory flow (PEF) between the groups. CONCLUSION: It is concluded that lung function in type 1 diabetic patients is impaired by a decrease in FVC and FEV1 as compared to their matched controls. Additionally, the years of disease showed a dose-response effect on lung function. PMID- 16311657 TI - Significance of anti-glomerular basement membrane antibodies in type 2 diabetic patients. AB - OBJECTIVE: Diabetic glomerulosclerosis is a common cause of proteinuria in patients with type 2 diabetes mellitus (DM), however, primary glomerulonephritis can superimpose on it. The aim of this study is to evaluate the significance of anti-glomerular basement membrane (GBM) antibodies in type 2 DM. METHODS: We choose 60 well known Saudi patients with type 2 DM diagnosed according to American Diabetes Association criteria attending the medical outpatient clinic at Al-Noor Specialist Hospital in Makkah, Kingdom of Saudi Arabia between July 2004 and December 2004, after written consent and divided them into 3 groups according to the degree of albuminuria, either normal, micro albuminuric or with diabetic nephropathy. We choose 20 healthy subjects as the control. We checked anti-GBM serum level by indirect solid phase immunosorbent assay (ELISA). RESULTS: There was significant increase in anti-GBM serum levels in diabetic patients compared with the control subjects, mainly in the normal and micro albuminuric patients groups, but not in patients with diabetic nephropathy. CONCLUSION: Anti-GBM antibodies are a significant predictor of diabetic nephropathy especially in the early phases, however, we need further studies to document its clinical importance in such common, important diabetic complications. PMID- 16311658 TI - The use of mitomycin-C for respiratory papillomas: clinical, histologic and biochemical correlation. AB - OBJECTIVE: To assess the clinical effects of mitomycin-C (MMC) on human papilloma virus (HPV)-infected tissues of the airway. METHODS: We included 10 patients with previous histologic diagnosis of recurrent respiratory papillomas (RRP) in this prospective study, conducted at the Department of Otolaryngology-Head and Neck Surgery, University of Alexandria, Egypt, between January 2000 and December 2002. Under general anesthesia, each patient underwent laser excision of all visible papillomas, followed by topical application of 1 cc of 0.5 mg/ml MMC. The procedure was repeated weekly until no visible papillomas could be microscopically detected. We histopathologically studied the obtained specimens and tested for the presence of HPV DNA using polymerase chain reaction (PCR) technique. We collected blood samples from all patients and from another 10 healthy volunteers for determination of serum interleukin-2 (IL-2) level using enzyme-linked immunosorbent assay technique. RESULTS: We achieved clinical remission in 8 of the patients (80%), a fact that was confirmed histopathologically and by PCR data. The mean serum IL-2 levels+/- SD was significantly lower in papilloma patients (83.6+/- 28.83 pg/ml) than in control subjects (196.3+/- 42.46 pg/ml) (p<0.01). Among patients with RRP, serum IL-2 levels+/-SD was lower in initial samples (83.6+/- 28.83) than follow-up (95.7+/- 27.98 and 112.3+/- 33.8 and 129.4+/- 34.04) and remission samples (154+/- 37.84 pg/ml) CONCLUSION: Our result suggests that topical application of MMC may act adjunctively to laser surgery for RRP. PMID- 16311659 TI - Recurrent laryngeal nerve injury and preservation in thyroidectomy. AB - OBJECTIVE: We aim to evaluate the rates of recurrent laryngeal nerve (RLN) injury after thyroidectomy and to put forward the factors influencing the risk of RLN injury during thyroid surgery. METHODS: We retrospectively analyzed the records of 418 patients who underwent thyroid surgery for thyroid disease at the Turkish State Railway Hospital, Ankara and Gazi University Hospital between 1989-2003 for RLN injury and factors affecting this complication. Moreover, we evaluated 6 different types of operations used during surgical practice. Indirect laryngoscopy was performed preoperatively and was repeated postoperatively for all patients. RESULTS: Three hundred and thirty-nine (81.1%) were females and 79 (18.9%) were male. Indications for surgery were multinodular goiter in 253 cases, solitary nodule in 69, hyperthyroidism in 68, thyroid carcinoma in 5 and recurrent goiter in 23 cases. Bilateral subtotal thyroidectomy was performed in 286 cases (68.4%), unilateral subtotal thyroidectomy in 52 (12.4%), unilateral total thyroidectomy in 25 (5.9%), bilateral total thyroidectomy in 22 (5.3%), nodule excision in 10 (2.4%) and completion thyroidectomy for recurrent goiter in 23 (5.5%) cases. Unilateral vocal cord problems occurred, 16 (3.8%) cases and in 6 (1.2%) cases it became permanent. The distribution of permanent RLN paralysis over the cases was 0.04% multinodular goiter, 2.9% hyperthyroidism and 8.7% recurrent goiter (p<0.05). Transient RLN paralysis rate was 2%, while permanent RLN paralysis rate was 0.03% for bilateral subtotal thyroidectomies. In addition, in unilateral total thyroidectomies, transient RLN paralysis was 12% while permanent paralysis was 4%. For bilateral total thyroidectomies, 13.6% was transient and 9% was permanent RLN paralysis and 13% was transient and 8.7% was permanent paralysis for completion cases (p<0.05). CONCLUSION: We can avoid RLN injury during thyroid surgery by identifying the nerve and following its course carefully. Intraparenchymal dissection or subtotal excision can be performed if failure to identify RLN occur, and new operative techniques and medical management of benign thyroid diseases should be considered. PMID- 16311660 TI - Reconstruction techniques in renal injury. AB - OBJECTIVE: Kidney repair reconstruction techniques are controversial. The conventional technique is suturing, but this is usually with further loss of viable tissue as it promotes scaring. In this animal model, we investigated the parenchymal effect of different sutures and methods. METHODS: We carried out this study in the year 2000 in the Animal Laboratory of Dicle University, Diyarbakir, Turkey. We used 40 New Zealand white rabbits in this study, randomized into 5 groups. We separated the left kidney from Gerota's fascia, and performed standard lacerations with incisions 5 mm deep and 10 mm in length. We performed no reconstruction procedure in group 1, and used homeostatic collagen powder in group 2. We primarily sutured the lacerations with chromic gut (4/0) in group 3, and sutured the kidney in group 4 with polyglactin (4/0). We wrapped the kidney with a polyglactin mesh in group 5. We sacrificed 2 rabbits in each group postoperatively on day 2, 15, 45 and 90, and performed left nephrectomy for histological investigation, and assessed interstitial inflammation. RESULTS: While group 5 established the best results, the other 4 groups had similar intermediate results. The pseudocapsule was visible macroscopically in the polyglactin mesh group. We could see mononuclear cell infiltration, dilatation of tubules, atrophy of tubules, and interstitial fibrosis in all groups except group 5. CONCLUSION: In this animal model, we found that the most appropriate repair material for kidney surgery was polyglactin mesh. PMID- 16311661 TI - The virulence factors and antibiotic sensitivities of Escherichia coli isolated from recurrent urinary tract infections. AB - OBJECTIVE: We determined the antibiotic sensitivities of uropathogenic Escherichia coli (UPEC) strains isolated from the urine of patients who have recurrent urinary tract infections (UTIs). METHODS: Our study was carried out between November 2000 and January 2002 at the Infectious Diseases Clinic, Istanbul Haydarpasa Numune Hospital, Istanbul, Turkey. We compared the virulence factors (fimbrial adhesion, hemolysin production, motility property) of 50 strains of Escherichia coli (E. coli) isolated from urine with the same properties of 25 strains of E. coli isolated from stool specimens of healthy individuals. In addition, we detected the virulence factors of UPEC strains using a microbiological and biochemical methods and by using disk diffusion method, we were able to investigate the sensitivity of the strains to the antimicrobials. RESULTS: We found the level of mannose-resistant (MR) fimbriae bearing in the UPEC strains to be significantly higher than that in the controls (odds ratio=10.27, p<0.001). The difference in mannose-resistant hemoagglutination (MRHA) and mannose sensitive hemoagglutination (MSHA) bearing levels in UPEC strains were rather high. This difference was regarded as significant in terms of showing the virulence of fimbriae bearing strains (odds ratio=29.03, p<0.001). CONCLUSION: Our study demonstrates that strains with MR fimbriae have a rather high virulence (p<0.001), and that a combination of MR+MS fimbriae increased that virulence (p<0.001). As MR strains have a greater adhesive property, the determination of MR fimbriae bearing as high shows that fimbriae bearing plays an important role in widespread and resistant strains, especially in recurrent UTIs such as in our study. In addition, hemolysin capability was also a virulence factor in recurrent UTIs (p<0.01). In addition, the sensitivity of the strains to the antimicrobials appeared in the following order; imipenem 93%, norfloxacin 89%, ciprofloxacin 85%, netilmicin 80%, amikacin 78%, ceftriaxone 74%, gentamicin 72%, nitrofurantoin 71%, ampicillin-sulbactam 60%, amoxicillin-clavulanate 58%, Trimethoprim/sulfamethoxazole 45%, ampicillin 35%. PMID- 16311662 TI - Etiology of childhood diarrhea in Zliten, Libya. AB - OBJECTIVE: To determine the etiological agents of diarrhea in children from a small semi-urban city in Libya and the association of age, gender, seasonal variation, breast-feeding, source of water for drinking, or antibiotic use with the isolation of enteropathogens and whether such agents are community or hospital acquired. METHODS: Using standard microbiological techniques we examined stool samples from 169 children (70 females) aged a few days to 12 years with acute diarrhea for viral, bacterial and parasitological agents. We used the disc diffusion method to determine the susceptibility of bacterial pathogens to antimicrobial agents. We carried out the study between April 2000 to March 2001. RESULTS: We detected a single agent in 44.4%, rotavirus in 26.6%, Salmonella in 13.6%, and Cryptosporidium in 13% of patients and other enteric pathogens, Shigella in 3.6%, Aeromonas in 5.5%, Entamoeba histolytica/dispar in 11.8, and Giardia lamblia in 1.2%. Serotyping of isolated Salmonella resulted in 21 being Salmonella enteric serotype heidelberg and 3 Salmonella enteritidis. We detected both serotypes in one child. More than 75% of the isolated Salmonella were resistant to 6 different commonly used antimicrobial agents. CONCLUSION: We found rotavirus, non-typhoid Salmonella and Cryptosporidium to be the most important enteric agents associated with childhood diarrhea in Zliten. The isolated bacterial pathogens showed high resistant rates, particularly among the Salmonella, to the commonly used antimicrobial agents. The ease of which one can obtain these drugs in Zliten may play a role in such resistance. PMID- 16311663 TI - Clinical sources and prevalence of resistance to antimicrobials of Klebsiella pneumoniae strains in Trinidad. AB - OBJECTIVE: To assess the frequency of Klebsiella pneumoniae (K. pneumoniae) recovery from clinical specimens from hospital and community patients, their antibiotic profile, the extent of extended-spectrum beta lactamase (ESBL) production among such isolates, and the impact on patient management. METHODS: The study recovered 1,118 K. pneumoniae strains from various clinical specimens from hospital and community patients, from 1995 to 2004, at the Eric Williams Medical Sciences Complex. All 1,118 isolates underwent testing for multi antibiotic resistance, of which 480 that fulfilled the criteria for possible ESBL production underwent further examination, according to guideline recommendations of the National Committee for Clinical Laboratory Standards. RESULTS: From 68,537 specimens processed, 62.9% were from hospital patients and 37.1% from community patients. Approximately 21% of the specimens yielded positive bacterial cultures from which, 1,118 were K. pneumoniae strains. Ciprofloxacin, imipenem, aztreonam, nalidixic acid and gentamicin showed the greatest efficacy (>95% sensitivity) against isolates from both hospital and community sources. Tetracycline and ampicillin showed almost 100% resistances. The other antibiotics displayed varying degrees of resistance. The prevalence of ESBL production was approximately 8.5% and most ESBL producers (51.2%) were from urine, followed by wounds (22.0%), blood (19.5%), and lower respiratory tract specimens (4.9%). Five K. pneumoniae isolates were resistant to the carbapenem, and imipenem. CONCLUSION: The study isolated K. pneumoniae from 8% of patients. All K. pneumoniae isolates were resistant to more than 2 antibiotics. The prevalence of ESBL production was 8.5%. Five (12.2%) strains of ESBL producers were resistant to imipenem. Continued infection control measures and prudent use of antibiotic agents are essential in reducing the spread of multi-resistant ESBL producing K. pneumoniae. PMID- 16311664 TI - Prenatal diagnosis of beta-thalassemia in the West Bank and Gaza. AB - OBJECTIVE: This study focuses on the genetic aspect of beta-thalassemia among 88 at risk couples from the West Bank and Gaza, and the attitude of these couples toward prenatal diagnosis and its outcome as a preventive method. METHODS: We tested 130 prenatal samples for beta-thalassemia during the period from January 1999 to July 2005. We performed prenatal diagnosis in these cases using the amplification refractory mutation system, as well as beta-globin gene sequencing as a conformational method. We drew a chorionic villus sample (CVS) for 1st trimester pregnant women and amniotic fluid (AF) for those in the 2nd trimester depending on the stage the pregnant woman contacted our lab. RESULTS: The DNA analysis of 130 prenatal samples revealed 25 (19.2%) cases of beta-thalassemia major and 67 (51.5%) cases of beta-thalassemia carriers, while the remaining 38 (29.2%) were normal. The 25 affected fetuses were aborted according to the wishes of the parents. In the tested 88 couples, 14 mutations of beta-thalassemia were identified. These mutations and their frequencies were: IVSI-110 (22.2%), IVSI-6 (13.6%), Cd37 (12%), IVSI-I (9.7%), IVSII-1 (6.2%), Cd39 (9%), Cd6 (sickle cell mutation) (8.5%), Cd5 (8%), Cd8/9 (2.8%), Cd106/107 (2.8%), -30 promoter (1.1%), 88 promoter (1.1%), IVSI (-1) (2.3%) and IVSI-5 (0.6%). We found that in 77.3% of the couples, both the mother and the father carry the same type of mutation while 22.7% of them carry different mutations. We found 77.9% consanguinity among the couples CONCLUSION: We found very good acceptability for prenatal diagnosis in beta-thalassemia afflicted families. All couples with affected fetuses opted for abortion. The spectrum of mutations in the tested couples revealed several similarities to neighboring countries with -88 promoter mutation reported for the first time in our region. PMID- 16311665 TI - Zinc deficiency among a healthy population in Baghdad, Iraq. AB - OBJECTIVE: To determine the prevalence of zinc deficiency and the current zinc status among a sample selected from the healthy population in Baghdad, Iraq. METHODS: We carried out a community-based study in Baghdad City, Iraq from November through June 2002. We selected a sample of 2090 healthy subjects (aged 1 month to 85 years). We used a pre-tested questionnaire, designed to obtain information on gender, birth dates, height, weight, residence, habitual food consumption patterns, and social status. We performed laboratory assessment of serum zinc level, dietary assessment of food frequency and usual zinc intake. We considered subjects with serum zinc concentration of <7.7 umol/l zinc deficient and >/-7.7 to 12.3 umol/l mild to moderately zinc deficient. RESULTS: The prevalence of zinc deficiency among the studied sample was 2.7%. We found mild to moderate zinc deficiency among 55.7% of the study sample. Dietary zinc intake assessment showed that 74.8% of the studied sample consumed less than the recommended intake, and in 62.3%, the intakes were deficient and grossly deficient. Mean daily zinc ranged from 5.2 mg in children to 8.5 mg in adults. CONCLUSION: We observed a high prevalence of mild to moderate zinc deficiency, with inadequate dietary zinc intake among a considerable proportion of the studied sample. Zinc supplementation may be an effective public health intervention means to improve the zinc status of the population. PMID- 16311666 TI - Dietary intake and nutritional status of Turkish pregnant women during Ramadan. AB - OBJECTIVE: To determine the effects of Ramadan fasting on dietary intakes, nutritional status and biochemical parameters of Turkish pregnant women. METHODS: We carried out this study at Dr. Zekai Tahir Burak Women's Health Care and Research Hospital, Ankara, Turkey from October 15th to November 13th 2004. Forty nine fasting group and 49 non-fasting group (control group) voluntarily enrolled for the study. We analyzed the blood biochemical parameters of pregnant women and obtained the dietary intakes from the groups. RESULTS: Compared to the control group, weight gain and energy intake (p<0.05 for second and third trimesters) was less in the fasting group. The percentage of protein (p<0.05 for first and second trimesters) and carbohydrates (p<0.05 for all trimesters) from total energy was higher in the fasting group than in the control group. We noticed a slight increase in the fasting blood glucose, serum total cholesterol high-density lipoprotein-cholesterol and triglycerides (p<0.05 for first trimester) concentrations in the fasting group during Ramadan. However, we found decreased levels in the plasma urea, total cholesterol, triglycerides, low-density lipoprotein-cholesterol and total protein and albumin levels of the fasting group in comparison with the control group. Dietary mineral and vitamin intakes were lower than the recommended daily allowance, except vitamin A and vitamin C in both groups. CONCLUSION: Based on the results of the present study, Ramadan fasting had no significant adverse effect on the health of pregnant women. PMID- 16311667 TI - Impact of health education on knowledge and attitudes of Saudi paramedical students toward HIV/AIDS. AB - OBJECTIVE: To assess the impact of health education on the knowledge and attitudes of paramedical students in Saudi Arabia toward HIV/AIDS. METHODS: We carried out an interventional study on a sample selected from students of health institutes and health colleges in Saudi Arabia during the calendar year 2002 2003. RESULTS: The intervention shows a positive effect on students' knowledge regarding means of transmission of HIV and means of protection from HIV/AIDS. Furthermore, it has a positive impact on students' attitudes toward accepting discussion of AIDS topics with others, acceptance of home care for HIV infected family member and acceptance of HIV-infected individual's right at work. Preference of confidentiality was also significantly increased after intervention. CONCLUSION: Health education intervention has a positive impact on students' knowledge and attitudes towards HIV/AIDS and we recommend a nationwide health education program on HIV/AIDS. PMID- 16311668 TI - Clinical characteristics of Crohn s disease in Qatar. AB - OBJECTIVE: To assess the clinical characteristics of Crohn's disease (CD) in Qatar. METHODS: This is a retrospective descriptive study. We studied 50 CD patients admitted to Hamad General Hospital over a period of 8 years from January 1995 to December 2002. We retrospectively collected and analyzed the demographic and clinical data. We made the diagnostic classification of definite CD in accordance with the criteria based on the International Classification of Disease ninth revision [ICD-9]. We based the obtained information on the following parameters: the age at the time of admission, gender and Crohn's risk factor profiles. RESULTS: Of the studied population, 60% were males and 40% females. Sixty-two percent were Qataris and 38% expatriates. Most of the subjects were <40 years of age (80%). Fifty-two percent had ileo-colonic disease, 24% colonic and 24% ileal disease. The majority of the Crohn's patients had the symptom of abdominal pain (84%), followed by weight loss (80%) and diarrhea (70%). Pallor sign was most frequent in patients with inflammatory CD (76.9%) and obstructive CD (66.7%), whereas we found the anal signs in patients with fistulas (84.2%). CONCLUSION: The study showed a relatively high frequency of ileo-colonic Crohn's disease compared to colonic and ileal diseases. We observed that the most frequent symptom is abdominal pain. PMID- 16311669 TI - Thrombolytic therapy in acute pulmonary embolism due to heparin-induced thrombocytopenia type II. AB - Hospitalized patients are in danger of deep venous thrombosis either due to a genetic tendency, immobilization or the underlying medical condition. Paradoxically heparin, the substance used to prevent this complication, can lead to thrombo-embolic phenomena, which can be life threatening. We report a case of heparin-induced thrombocytopenia, which caused a massive pulmonary embolism, and its management by administering a thrombolytic agent in a situation where bleeding seems inevitable. PMID- 16311670 TI - Cardiac echinococcosis: echocardiographic diagnosis with a fatal clinical outcome. AB - Hydatid disease HD is an endemic disease found in various regions of the world. The organs mostly affected are the liver and lung. Cardiac involvement in HD is rare. The symptoms are mainly due to the mass effect of the calcific hydatid cyst obstructing the blood or the lymphatic vessels. Other manifestations are secondary infection or cyst rupture in the involved organ. Here, we report a middle aged female patient with no history of medical illness who presented to the emergency room with an unrecordable blood pressure. Echocardiogram showed multiple calcific cysts of echinococcosis granulosa in the left ventricle cavity. In the clinical setting, where there is detection of HD elsewhere in the body, an echocardiogram is mandatory as cardiac involvement is serious and may be fatal. PMID- 16311671 TI - Essential thrombocythemia in a child: diagnostic and therapeutic dilemma. AB - We report an 11-year-old child with essential thrombocythemia ET, a very rare myeloproliferative disorder among children. Essential thrombocythemia can be complicated by life-threatening thrombosis with a risk of converting into acute leukemia. Cytoreductive therapy may reduce the risk of thromboembolic complications. We usually recommend cytoreductive treatment for asymptomatic adult patients with platelet counts of more than 1.5 million/micro liter, but treatment remains obscure in children. Herein, we report the results of child with ET, treated successfully with hydroxuea. PMID- 16311672 TI - Rare variation of the celiac trunk and related review. AB - In this study, we report a rare variation of the branching of the celiac trunk. During a routine abdominal dissection on a female cadaver, we found the celiac trunk to emerge from the abdominal aorta as 2 roots named hepatogastric trunk and hepatosplenic trunk. The hepatogastric trunk arises from the anterior surface of the abdominal aorta and divides into an aberrant branch to the right lobe of the liver, a branch to the right hemi diaphragm, the left hepatic and the left gastric arteries. The hepatosplenic trunk, which arises 1.5 cm below the hepatogastric trunk, gave off the common hepatic and splenic arteries. The common hepatic artery divided into the gastroduodenal, the right branch to the hepatic and the cystic arteries. It is important to know the variations of hepatogastric trunk and hepatosplenic trunk for the success of surgical operations to the liver and radiological investigations of those regions. PMID- 16311673 TI - Hip arthroplasty for ochronosis. AB - Alkaptonuria is a metabolic disorder in which homogentisic acid oxidase is absent. Therefore, homogentisic acid accumulates in cartilage and connective tissues. We can diagnose ochronotic arthropathy, a manifestation of long standing alkaptonuria, through careful radiological, physical, and laboratory examination. In this report, we describe 4 cases of ochronotic arthropathy to which we applied cementless total hip prosthesis due to severe hip involvement. PMID- 16311674 TI - Recurrent maturing perineal lipoblastoma. AB - Lipoblastoma is a rare benign neoplasm of fetal adipose tissue that we see mostly in infants and young children less than 3 years of age. Most lipoblastomas occur on the extremities, trunk, head and neck, and various other organs have been described. We report a case of a recurrent perineal lipoblastoma in a 2.5-year old boy, which showed maturation of the lipoblasts as compared to the primary tumor. PMID- 16311675 TI - Hemolytic uremic syndrome associated with Streptococcus pneumoniae. PMID- 16311676 TI - Cardiotoxicity resembling myocardial infarction that occurs during chemotherapy with 5-fluorouracil. PMID- 16311677 TI - Albumin: its place in critical care practice. PMID- 16311678 TI - Use of adrenaline as an adjunct to local anesthetic agent: a cause of concern for the anesthesiologist. PMID- 16311679 TI - Spontaneous rupture of the ovarian artery following spontaneous vaginal birth. PMID- 16311680 TI - Two anatomic variations in the arm related to the median nerve. PMID- 16311681 TI - Sex determination using sex determining region Y primers by single conventional polymerase chain reaction. PMID- 16311682 TI - Serum calcium and phosphorus levels in patients with ischemic heart disease. PMID- 16311683 TI - Blood and saliva lactate levels during recovery from supramaximal exercise. PMID- 16311684 TI - Target level controlled sedation: an alternative to general anesthesia in endovascular treatment of intracranial aneurysms. PMID- 16311685 TI - The frequency of skin diseases in obese children and adult Iraqi population. PMID- 16311686 TI - Prescribing pattern of angiotensin- converting enzyme inhibitors and beta blockers after acute myocardial infarction. PMID- 16311687 TI - Metastatic malignancies of unknown primary site and malignancies of unknown origin whether primary or metastatic. PMID- 16311688 TI - Prevalence of factor V Leiden and prothrombin G20210A gene mutation. PMID- 16311689 TI - [Consensus: Rational approach towards the patient with cancer, fever and neutropenia]. AB - The severity and duration of post chemotherapy neutropenia were recognized during the 1960s as main predisposing factors for infections in cancer patients. At the beginning of the 70's a standard management approach for all febrile neutropenia (FN) episodes was proposed, based on hospitalization and intravenous empirical broad spectrum antibiotic therapy. Widespread use of this approach resulted in a significant reduction in mortality attributable to bacterial infections. During the last 10 to 15 years, reappraisal of this standard approach has been done by several research groups who question the benefit of treating all FN patients similarly without taking in to consideration differences in severity of the FN episodes. This reappraisal has led during the 1990s to the development of the concept of high and low risk FN episodes that has been the base for the adoption of selective therapies based on the risk categorization of the individual patient. The Chilean Infectious Diseases Society called upon two government National Programs responsible for the appropriate distribution of chemotherapeutic drugs to all pediatric and adults cancer patients within the public health system, and upon the Chilean Hematology Society for the development of a Consensus on Diagnosis, Treatment and Prevention of Infections during FN Episodes in Cancer patients. The need for this Consensus is based on two main aspects: the new approaches proposed during the past year for management of these episodes, and the increasing population of cancer patients receiving improved chemotherapeutic agents that has increased there survival possibilities as well as there possibility to suffer a FN episode. The topics discussed in this document are based on an updated systematic and analytic review of the medical literature including epidemiology, laboratory diagnostics, risk categorization, treatment and prophylaxis. National data was included when available in order to provide the healthcare personnel that take care of these patients with best evidence based recommendations adjusted to the Chilean reality. PMID- 16311690 TI - [Chronicle of an announced pandemic?]. AB - In the last years our country has been affected by several outbreaks of infectious diseases such as Cholera and Hanta virus and recently, by pathogens associated to red tide. Chile was able to manage those emergencies using the local health system. The new threat that may emerge and could eventually overcome that capacity, is the possible H5N1 influenza virus outbreak. Influenza is responsible for the most destructive pandemic, the Spanish influenza, that killed over 40 million individuals in 1918. The new influenza strain (H5N1) is at present endemic in poultry in Asia and has been associated to human fatal cases in Hong Kong and Vietnam. Even though this strain is not able yet to be transmitted among humans, evidence has accumulated that such ability could be reached by the new strain, since it was already detected in pigs. That particular evidence may indicate that the virus could adapt to infect humans, since a similar situation was observed in several of the influenza pandemics. The World Health Organization set a "task force" to develop a strategy that may help to control the virus spread. Several countries are already stocking anti-flu drugs and others are developing new vaccine that are currently been assayed in human volunteers. It is possible that we may have a vaccine before the outbreak; this development is even faster than for SARS. The mayor question to be addressed for developing countries is: what will be done if we do not have the vaccine on time? PMID- 16311691 TI - [Predictors of non fatal cardiovascular events in a Chilean cohort: results of the San Francisco Project]. AB - BACKGROUND: The San Francisco Project is a prospective study aiming to determine the most important cardiovascular risk factors in the Chilean population. AIM: To report the results of the San Francisco Project after a mean follow up of five years. MATERIAL AND METHODS: Between 1997 and 1999, 920 subjects (aged 39.5 +/- 16.3 years, 382 males), free of cardiovascular events, were examined to determine their cardiovascular risk factors. The prevalence of risk factors was compared with the Chilean National Health Survey. All non fatal cardiovascular events were registered until 2003. A logistic regression model including gender, obesity, family history, educational level and classical cardiovascular risk factors, was performed to identify independent predictors of cardiovascular events. RESULTS: The prevalence of cardiovascular risk factors in the studied population was similar to that of the Chilean National Health Survey, except for serum lipid levels, that were lower in the latter. The mean follow up was 5.3 +/- 0.3 years (range 4-6 years), and 26 cardiovascular events were registered. The independent predictors of these events were age, with a relative risk (RR) of 4.3 and 95% confidence intervals (CI) of 1.9 to 9.5 (p <0.001), high blood pressure with a RR of 5.2 (95% CI 2.3-11.7, p <0.001), diabetes mellitus with a RR of 4.5 (95% CI 1.9-10.7, p <0.01) and a low socioeconomic level with a RR of 3.5 (95% CI 1.5 7.9, p <0.01). The model did not accept serum lipid levels, smoking and alcohol intake as independent predictors. CONCLUSIONS: In this cohort of Chilean people, the independent predictors of non fatal cardiovascular events were high blood pressure, diabetes mellitus and a low socioeconomic level. PMID- 16311692 TI - [Evolution of the nutritional status of six years old Chilean children (1987 2003)]. AB - BACKGROUND: As a consequence of the epidemiological transition in Chile, the nutritional status of the population has changed notoriously. AIM: To study the changes in the nutritional status of six years-old Chilean children from 1987 to 2003. MATERIAL AND METHODS: The computer data base of a government institution in charge of the School Lunch Program (JUNAEB) was used. It contains data on weight, height, sex and date of birth of six years old children, in the years 1987, 1990, 1993, 1996, 2000, 2001, 2002 and 2003. RESULTS: The data of approximately 80,000 children was analyzed. Both weight and height increased over the study years. There was a significant reduction in the number of children with weight deficit but also an increase in the proportion of children with obesity. Stunting was also reduced. The proportion of children with HAZ over 2 SD increased from 0.76 in 1987 to 2.2% in 2003. The prevalence of obesity has not increased since the year 2000. CONCLUSION: The stabilization in the prevalence of obesity is a positive finding, considering its adverse consequences for health. PMID- 16311693 TI - [Depressive symptoms after an acute myocardial infarction]. AB - BACKGROUND: Depression after myocardial infarction (MI) is a frequent disorder and it increases the long-term risk of cardiac mortality. AIM: To assess the frequency of depressive symptoms and the history of depression in hospitalized post-MI patients. PATIENTS AND METHODS: During three months, depressive symptoms and history of depression were studied in 47 consecutive patients (mean age 59.8+/-9.5 years, 68% male), admitted for MI to the Barros Luco Trudeau Hospital. The Beck Depression Inventory (BDI) and the Composite International Diagnostic Interview (CIDI 2.1) were used with DSM-IV diagnosis criteria. RESULTS: According to the results obtained using the CIDI, 27,7% of the patients had a history of depression. This occurred in 53,3% of women and 15,6% of men (p <0.01). During the hospitalization, 38.3% of patients had depressive symptoms (BDI > or =17 points), affecting 60% of women and 28,1% of men (p <0.02). In women and patients with history of depression, depressive symptoms tended to be more common and more severe. CONCLUSIONS: Depressive symptoms in post-MI patients are frequent and attending physicians should actively detect them. PMID- 16311695 TI - [Indications and long term results of elective surgery for sigmoid diverticular disease]. AB - BACKGROUND: There are no clear guidelines for the indication of elective surgery in sigmoid diverticular disease. AIM: To analyze the indications and long term results of elective surgery in sigmoid diverticular disease. MATERIAL AND METHODS: Retrospective review of 100 patients (age range 25-86 years, 51 male) with sigmoid diverticular disease, operated in a lapse of 22 years. Sixty seven patients answered a survey about their disease at the end of follow up. RESULTS: Among patients aged more than 70 years, there was a higher proportion of women. The main indication for surgery was recurrent diverticulitis in 54 patients, followed by diverticular fistula in 19. A sigmoidectomy was performed in 91 patients. Stapled anastomosis was performed in half of these patients. No patient died or required reoperation in the immediate postoperative period. During a follow up ranging from 8 to 280 months, 28 patients died for causes not associated with diverticular disease and five were lost. Those patients that answered the survey were free of symptoms related to diverticular disease and did not require new operations. CONCLUSIONS: In patients with sigmoid diverticular disease and recurrent diverticulitis or with fistulae, the long term results of surgery are satisfactory. PMID- 16311694 TI - [Prophylactic thyroidectomy in children and young people with hereditary medullary thyroid carcinoma: a Chilean experience]. AB - BACKGROUND: With the availability of the RET proto-oncogene genetic testing, it is possible to perform prophylactic total thyroidectomy among carriers of RET mutation. AIM: To evaluate the histological findings and the effects of the prophylactic total thyroidectomy in first-degree relatives of Chilean patients with multiple endocrine neoplasia type 2 (MEN 2) based on the Ret proto-oncogene analysis. SUBJECTS AND METHODS: Nineteen patients belonging to 11 MEN 2 families underwent total thyroidectomy. Of these, 16 either with C cell hyperplasia (CCH) or microscopic medullary thyroid carcinoma (MTC) were selected for the final analysis. RESULTS: The age at the moment of thyroidectomy ranged from 3 to 24 years (median 9.5). The most common mutation was located in codon 634 (69%) followed by codon 620 (25%). Histopathology revealed MTC in 13 patients (81%, youngest 3 years, oldest ones 19 and 24 years) and CCH in 3. A significant correlation was observed between basal preoperative serum calcitonin/tumor size (r = 0.53, P < 0.05) and age/tumor size (r = 0.56, P < 0.03), but not between basal preoperative serum calcitonin and age. Stimulated preoperative calcitonin levels were confounding and not useful for differentiating CCH from MTC. None of patients in whom cervical dissection was done (9/16) presented lymph node metastases, including the oldest ones. All patients but the older ones were biochemically cured after a mean of 5 years of follow-up. CONCLUSION: Prophylactic total thyroidectomy should be done early in life because there is an age-dependent progression from HCC to MTC. MTC often precedes biochemical detection of the disease. PMID- 16311696 TI - [Treatment of Familial Adenomatous Polyposis and family screening]. AB - BACKGROUND: To reduce the mortality associated to Familial Adenomatous Polyposis (FAP), screening of close relatives of patients with the disease is crucial. AIM: To analyze the results of the surgical treatment of patients with FAP, and to evaluate the family screening. PATIENTS AND METHODS: Clinical records of patients operated in our institution since 1977, were reviewed analyzing surgical and pathological results, and follow up. In their family members, we evaluated and analyzed the performance of screening tests, former surgeries, history of disease related cancer and mortality, all due to FAP. RESULTS: Between January 1977 and August 2002, 15 patients were operated on. Of these, only 33% consulted on the setting of a familial screening. A proctocolectomy and terminal ileostomy was performed in 27% of patients; 20% had a proctocolectomy and ileal pouch, and 53% underwent a total colectomy with ileo-rectal anastomosis. Morbidity and mortality were 7% and 0%, respectively. Twenty percent had a colorectal cancer. During a median of 68 months follow-up, the disease-related survival was 92%; no cancer of the rectal stump was detected. Of the 122 family members identified, only 33% with clear indication of screening underwent a colonoscopy. Twenty-nine percent had a confirmed FAP and were operated: in 61% of them a colorectal cancer was found, and 91% of these died. CONCLUSIONS: The results of the surgical treatment of FAP are satisfactory. Nevertheless, family screening should be improved to reduce the high rates of mortality revealed in the study of other family members. PMID- 16311697 TI - [Possible association due to linkage disequilibrium of TGFA, RARA and BCL3 with nonsyndromic cleft lip with or without cleft palate in the Chilean population]. AB - BACKGROUND: Nonsyndromic cleft lip/palate (NSCLP) is a congenital malformation with the characteristics of a complex genetic trait. Based on experimental evidences as well as on association and linkage studies candidate genes TGFA, RARA and BCL3 have been postulated as being involved in the genetic etiology of this pathology. AIM: To test the possible association due to linkage disequilibrium between microsatellite markers located at less than 1cM from the three candidate genes and nonsyndromic cleft lip/palate using the case-parents trio design. PATIENTS AND METHODS: The sample consisted of 58 case-parents trios. Two microsatellite markers, flanking each one of the candidate genes were analyzed by means of the polymerase chain reaction (PCR) with fluorescent labeled microsatellite markers. Electrophoresis of the PCR products was performed on a laser-fluorescent automatic DNA sequencer. Nonparametric ETDT was used to analyze the genotype data. RESULTS: Significant linkage disequilibrium was detected between D2S443 (TGFA) and NSCLP. Significance was almost reached between D17S800 (RARA) and NSCLP. Alleles 239bp (D2S443) and 172bp (D17S800) showed significant preferential transmission from heterozygous parents to affected offspring. In the case of BCL3 both markers showed no significant results. CONCLUSIONS: The results of the present study do not show clear evidence that TGFA or RARA could be involved in the genetic etiology of NSCLP. Even though the importance of retinoic acid in the development of the embryo is well documented the results obtained for RARA are difficult to analyze. In relation to the possible role of BCL3 in NSCLP, recent information postulates that other genes located in the same chromosome region could be involved in NSCLP. PMID- 16311698 TI - [Detection of human metapneumovirus in children hospitalized for acute lower respiratory infection in Santiago, Chile]. AB - BACKGROUND: Human metapneumovirus (hMPV) has recently been described as a new causal agent of acute low respiratory infection (ALRI) in children. In South America, detection has been reported only in Brazil and Argentina. AIM: To detect hMPV in children hospitalized for ALRI in Santiago, Chile. MATERIAL AND METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect N gene of hMPV in 182 nasopharyngeal aspirates that were negative for common respiratory viruses, obtained from children hospitalized for ALRI during 2003. RESULTS: Ten samples (5.4%) were positive, most of them detected during spring months. CONCLUSIONS: Since hMPV was detected among infants with ALRI in Santiago, further studies on their prevalence should be done in South America. PMID- 16311699 TI - [Extracorporeal membrane oxygenation (ECMO): consolidation of a neonatal pediatric program in Chile and report of three cases]. AB - Extracorporeal membrane oxygenation (ECMO) improves survival in neonatal and pediatric patients with reversible severe respiratory or cardiac failure, in whom intensive treatment fails. Since 1999, a multidisciplinary team is trained to form the first neonatal-pediatric ECMO center in Chile, according to the norms of the Extracorporeal Life Support Organization (ELSO). During 2003 the first three patients were admitted to the program: a male newborn with pulmonary hypertension, a 38 days old female operated for a total anomalous pulmonary venous connection and a 3 months old male with a severe pneumonia caused by respiratory syncytial virus. They remained in ECMO for five, seven and nine days respectively and all survived to the procedure. No neurological complications were observed after one and a half year of follow up. This consolidates the first national neonatal-pediatric ECMO program, associated to ELSO. Up to date, twelve patients have been admitted to the program. PMID- 16311700 TI - [Recurrent Impetigo Herpetiformis: successfully managed with ciclosporine. Report of one case]. AB - Impetigo Herpetiformis is a high-risk gestational skin disease that represents a risk for both the mother and offspring. Its management is based on multisystemic support and maternal steroid therapy. When these measures are insufficient to control the disease, the association of ciclosporine to the treatment has been proposed. We report a 24 year-old woman with a 16 weeks pregnancy, that presented with Impetigo Herpetiformis. The disease was refractory to the use of steroids, the patient had a metabolic decompensation and a dehydration with electrolyte imbalance. Therefore, treatment with ciclosporine was initiated and a rapid regression of the lesions was observed. Gestation was maintained, with a good perinatal outcome. PMID- 16311701 TI - [The risk of congenital malformations and genomic imprinting defects in assisted reproductive technologies and nuclear transfer cloning]. AB - Recent studies show that assisted reproductive technologies (ART), whether in vitro fertilization (IVF) or intra-cytoplasmatic sperm injection (ICSI) or applied to cloning by somatic cell nuclear transfer (SCNT) are associated to a higher risk of congenital malformations and errors in deprogramming, maintenance or reprogramming genomic imprinting in humans and animals. IVF and ICSI are also associated to an increased admission to neonatal intensive care units and more need for health care resources in infancy. A mutagenic effect of a chemical used in SCNT has been reported and gene depression was found in bovine embryos obtained by IVF or SCNT. The causes of these anomalies could be pathological conditions for which ART is applied, a direct effect of technologies on the zygotes or embryos, avoidance for zygotes or embryos of the oviduct path that is needed to elicit necessary immunity or genomic programming processes, or adaptive selective steps acquired during thousands of millions of generations in evolution. The knowledge of evolution is emphasized as essential in the scientific ethical analysis. PMID- 16311702 TI - [Vibrio parahaemolyticus infections and algal intoxications as emergent public health problems in Chile]. AB - There is interest in the paradigm that relates environmental sea changes to the emergence of diseases that affect both aquatic organisms in the sea and human beings. The emergence of Vibrio parahaemolyticus as an important cause of epidemic summer diarrhea in 2004 and 2005, confined mainly to the tenth region in Chile, could be a manifestation of this trend. This and other areas of the country have also experienced several outbreaks of paralytic shellfish poisoning (PSP), diarrheal shellfish poisoning (DSP) and amnesic shellfish poisoning (ASP) caused by harmful algal blooms (HAB) of Alexandrium catenella, Dinophysis acuta and Pseudonitzchia species, respectively. The short historical record of these pathological phenomena in Chile suggests that they are increasing in frequency and expanding their geographical range. The V parahaemolyticus isolates responsible for the Chilean outbreaks correspond mainly to the pandemic strain O3:K6. HAB found in Chile and the intoxications caused by them have similar biological characteristics to those described in other areas of the world. The tenth region, the area where these problems are emerging, produces approximately 80-90% of the shellfish consumed in Chile and a large proportion of the shellfish that is exported. Prevention of these public health problems can be attained by developing policies that increase environmental surveillance for Vibrios and toxic algae, improve the epidemiological surveillance of acute diarrhea and algal intoxications after the ingestion of raw bivalves, and educate the population on the mode of transmission of these diseases. Scientific capacity and laboratories need to be developed to widen the limited knowledge of the biology of Vibrio and toxic algae and the environmental factors that favor their emergence as public health and economic problems in Chile. PMID- 16311703 TI - [The influence of -238 and -308 TNF alpha polymorphisms on the pathogenesis and response to treatment in rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a systemic autoimmune disease that affects 0.8% of the world population, it affects the synovial membrane of joints and the clinical presentation encompasses a wide spectrum, ranging from a mild to a severe and erosive disease that causes joint and cartilage destruction which finally provokes irreversible structural damage and patient disability. In the last years, there have been important advances in the pathogenesis of this disease, the efforts have been concentrated on pro-inflammatory cytokines such as tumor necrosis factor alpha (TNFalpha). This protein guides numerous events in the synovial and systemic inflammatory process and is encoded in the Major Histocompatibility Complex (MHC), one of the most polymorphic of the genome. Polymorphisms affecting the TNFalpha gene and its regulatory regions are associated with RA prevalence and course. There is a possible association between these polymorphisms and the clinical response to the use of monoclonal antibodies anti-TNFalpha. The possibility that the determination of genotypes -238 and -308 may have prognostic and therapeutic consequences is debated nowadays. PMID- 16311704 TI - [Geometry of living systems and its importance in Medicine]. AB - We analyze the most relevant concepts about complexity of sciences, showing examples and consequences in medical sciences. In the first section we introduce some basic concepts about fractals and its relationship with the geometry of nature and chaos dynamics. We also introduce notions of critic systems, the role of energy in them and we propose a conceptual model for living systems. In this model we identify the non-linear dynamics as the dynamics of the living forms and fractals as the form resulting from that dynamics. In the second section we review medical and biological examples: infectious diseases, cardiovascular and behavior dynamics, from which useful concepts and medical hypotheses emerge. One of the most important concepts, is the proposition that homeostasis would have an irregular dynamic. Then, we could expect that healthy individuals should be characterized by irregular dynamics, but old or sick individuals would change, probably decreasing the complexity of its dynamics. Therefore, we ask ourselves if the use of therapies to obtain a constant internal medium can be a mistake. PMID- 16311705 TI - [Genomic medicine: a privilege of developed countries?]. AB - Genomic medicine is defined as the application of the knowledge originating from the Human Genome Project in health sciences. It has a vertiginous advance in developed countries, but its use in developing regions is scanty. This contributes to global health inequalities, stressing one of the greatest ethical problems that mankind is facing. In a study leaded by the University of Toronto Center of Bioethics, 28 highly qualified scientists elaborated a list of the ten most important biotechnologies to improve public health in developing countries. Several of these technologies are successfully applied in countries such as China, Cuba and Mexico. The factors that are hindering the application of such technologies in Chile, are briefly discussed. PMID- 16311706 TI - [Confidence intervals]. PMID- 16311707 TI - [Critical appraisal: combined T3 and T4 replacement therapy is not better than replacement with T4 alone]. AB - Conflicting results have recently been published about the benefits of combined thyroxine (T4) and triiodothyronine (T3) in treating hypothyroid patients. However these studies may have been underpowered to detect differences in psychological well-being specifically related to thyroxine replacement. We conducted a large, double-blind, randomized controlled trial of partial substitution of 50 microg of T4 by 10 microg of T3 (T3) vs placebo (T4 alone - 50 microg of T4 replaced) in 697 hypothyroid patients. Thyroid function showed a rise in the TSH (132%), a fall in Free T4 (35%, P <0.001) and unchanged basal Free T3 levels (P=0.92). At 3 months there was a large (39%) <> improvement in <> defined by the General Health Questionnaire 12 score (GHQ 12) in the control group compared with baseline and this was sustained at 12 months. Differences vs the intervention (T3) group were more modest with improvements in GHQ caseness (OR - 0.61; 95% CI: 0.42, 0.90; P=0.01) and HADS anxiety scores at 3 months (P <0.03) but not GHQ Likert scores, HADS depression, thyroid symptoms or visual analog scales of mood and the initial differences were lost at 12 months. These results may be consistent with a subgroup of patients showing transient improvement following partial substitution with T3 but do not provide conclusive evidence of specific benefit from partial substitution of T4 by T3 in patients on thyroxine replacement. They also emphasize the large and sustained <> that can follow changes in thyroid hormone administration. PMID- 16311708 TI - [UNESCO universal statement on bioethics and human rights]. PMID- 16311709 TI - [Therapy of hormone-refractory prostate cancer]. AB - PSA-progression following primary ADT defines an androgen-refractory but still hormone sensitive PCA which might respond to secondary hormonal manipulations. Secondary hormonal manipulations will result in a PSA decline >50% in about 60 80% of the patients with a mean duration of 7-17 months depending on the type of treatment. PSA-progression following secondary endocrine treatment defines hormone-refractory prostate cancer (HRPCA) which might be treated by systemic chemotherapy. Based on the results of 2 prospective, randomized clinical phase III trials comparing docetaxel and mitoxantrone, docetaxel results in a statistically significant survival benefit of 2.5 months, a significantly higher PSA- and pain response and represents the treatment of choice in the management of HRPCA. Bisphosphonates such as zoledronate represent another cornerstone in the management of PSA-progressive PCA demonstrating a significant benefit with regard to the prevention of skeletal related events. Furthermore, bisphosphonates might be indicated in the treatment of symptomatic bone pain. The current article critically reflects the various therapeutic options in the management of PSA progression following primary androgen deprivation for advanced prostate cancer. PMID- 16311710 TI - High-resolution SNP map of ASPN, a susceptibility gene for osteoarthritis. AB - Osteoarthritis (OA) is a very common bone and joint disease characterized by breakdown of cartilage in the joint. We recently found that an aspartic-acid repeat polymorphism of the asporin gene (ASPN) on chromosome 9 is associated with susceptibility to OA in Japanese. We provide here a high-resolution single nucleotide polymorphism (SNP) map within a 33.4-kb genomic region containing ASPN. A total of 19 SNPs were isolated from the region by systematic screening using 48 Japanese patients with OA: 7 SNPs in the 5' flanking region, 8 in introns, and 4 in the 3' untranslated region. Nine SNPs were novel. This high resolution SNP map will be a useful resource for analyzing genes associated with OA and other bone and joint diseases. PMID- 16311711 TI - Recent advances in the molecular basis of Lafora's progressive myoclonus epilepsy. AB - Lafora's disease (LD) is an autosomal recessive and fatal form of progressive myoclonus epilepsy with onset in late childhood or adolescence. LD is characterised by the presence of intracellular polyglucosan inclusions, called Lafora bodies, in tissues including the brain, liver and skin. Patients have progressive neurologic deterioration, leading to death within 10 years of onset. No preventive or curative treatment is available for LD. At least three genes underlie LD, of which two have been isolated and mutations characterised: EPM2A and NHLRC1. The EPM2A gene product laforin is a protein phosphatase while the NHLRC1 gene product malin is an E3 ubiquitin ligase that ubiquitinates and promotes the degradation of laforin. Analyses of the structure and function of these gene products suggest defects in post-translational modification of proteins as the common mechanism that leads to the formation of Lafora inclusion bodies, neurodegeneration and the epileptic phenotype of LD. In this review, we summarise the available information on the genetic basis of LD, and correlate these advances with the rapidly expanding information about the mechanisms of LD gained from studies on both cell biological and animal models. Finally, we also discuss a possible mechanism to explain the locus heterogeneity observed in LD. PMID- 16311712 TI - A transvaginal ultrasound study comparing transobturator tape and tension-free vaginal tape after surgical treatment of female stress urinary incontinence. AB - This study was carried out to compare ultrasonographic findings on patients after transobturator tape (TOT) and tension-free vaginal tape (TVT) procedures to evaluate displacement of tapes up to a 2-year follow-up and to test the correlation between bladder outlet obstruction and the tape position. Forty-nine patients had a transvaginal ultrasonographic evaluation after TOT (n=31) or TVT (n=18) procedures. Twenty-one patients from the TOT group and 12 from the TVT group had ultrasonographic evaluation both at 1 and 2 years follow-up. Tape position was evaluated with a complete sagittal view, and the width of the tape and the distance between the middle of the tape and the bladder neck were measured. Statistical analysis was performed using a t test. After clinical evaluation, stress test, and uroflowmetry with residual measurement, the tapes were compared on ultrasound with regard to clinical and urodynamic results: success in stress incontinence vs failure, bladder outlet obstruction vs no obstruction, and de novo urgencies vs no urgencies. No difference was found between TOT and TVT in the midurethral tape placement. The distance between the middle of the tape and the bladder neck at rest was similar at both 1 and 2 years after both TOT and TVT and was unchanged after coughing or Valsalva. The width of the tape was similar after TOT and TVT at both 1 and 2 years after the procedure. There was no difference in the distance between the middle of the tape and the bladder neck between cured patients, failure, bladder outlet obstruction, and de novo urgencies after both TOT and TVT. Transvaginal ultrasound examination showed no significant difference in the tape position between TOT and TVT. No correlation was found between ultrasonographic findings and postoperative voiding troubles for both techniques. PMID- 16311713 TI - C1q complement component and -antibodies reflect SLE activity and kidney involvement. AB - The role of the complement system in the pathogenesis of systemic diseases is very ambivalent. In systemic lupus erythematosus (SLE), many abnormalities in the activation of the complement system have been reported. The most important antibodies formed against the complement system in SLE are the ones associated with the C1q component. The aim of this study was to assess separately the anti C1q antibodies and C1q component in the serum from 65 patients with SLE, then in individuals with (n=33) and without (n=32) lupus nephritis and with active (n=36) and nonactive (n=29) form of the disease (European Consensus Lupus Activity Measurement, ECLAM>3, ECLAM90%. Depletion of CD8+ T cells has no effect upon therapy in the active treatment model, whereas depletion of CD4+ T cells completely abrogates anti-tumor activity. In a prophylactic setting, depletion of CD4+ and CD8+ T cells after the induction of a h5T4 immune response has no deleterious effect on protection following challenge with CT26-h5T4. In light of these studies, the role of antibodies in protection against tumor challenge was investigated. 5T4 specific polyclonal serum decreased tumor burden by approximately 70%. Thus, we conclude that CD4+ T cells are essential for the induction of a protective immune response and that antibodies are the likely effector moiety in this xenogeneic murine tumor model. PMID- 16311731 TI - Lentiviral vector expression of tumour antigens in dendritic cells as an immunotherapeutic strategy. AB - Therapeutic cancer vaccines need to stimulate a refractory immune system to make an effective anti-tumour response. We have explored the use of lentiviral vectors to deliver tumour antigen genes to dendritic cells (DC) as a possible mechanism of immune stimulation. Direct injection of a lentiviral vector encoding the melanoma antigen NY-ESO-1 in HLA-A2 transgenic mice primed NY-ESO-1-specific CD8+ cells that could be expanded by boosting with an NY-ESO-1 vaccinia virus. The expanded cells could kill NY-ESO-1(157-165) peptide-pulsed targets in vivo. In order to examine the priming step directly, we constructed another lentiviral vector expressing the melanoma antigen Melan-A (MART-1). Here we show that Melan A protein is also efficiently expressed after transduction of human DC cultured from peripheral blood mononuclear cells. When these transduced DC are co-cultured with autologous naive T cells, they cause the expansion of cells that recognise the HLA-A2 restricted Melan-A(27-35) epitope. The expanded cells are functional in that they release IFN-gamma upon antigen stimulation. Melan-A lentiviral vector transduced DC caused a similar level of naive T-cell expansion to Melan A(27-35) peptide-pulsed DC in four experiments using different HLA-A2 positive donors. These data suggest that a vaccine based either on DC transduced with a lentiviral vector ex vivo, or on direct lentiviral vector injection, should be assessed in a phase I clinical trial. PMID- 16311732 TI - Insufficient ability of omental milky spots to prevent peritoneal tumor outgrowth supports omentectomy in minimal residual disease. AB - BACKGROUND: The greater omentum is frequently involved in the course of gastrointestinal and ovarian tumors. Therefore, common practice in surgical treatment for especially gastric and ovarian cancer includes removal of the greater omentum. Paradoxically, many immune cells, such as macrophages that accumulate in so-called milky spots, reside within the omentum and are cytotoxic against tumor cells ex vivo. Consequently, omental macrophages might play an important role in killing tumor cells, and may hereby prevent development into local peritoneal recurrences. In the present study, we therefore evaluated the role of the omentum and the clinical relevance of omentectomy in minimal residual disease (MRD). METHODS: Tumor cell dissemination patterns on the omentum in a rat model were examined using DiI-labelled CC531s tumor cells. Additionally, intra peritoneal (i.p.) tumor load was investigated in rats that underwent omentectomy or sham laparotomy followed by i.p. injection of CC531s cells on day 21, which represented MRD. RESULTS: At 4 h post injection, tumor cells predominantly adhered on milky spots. Number of cells thereafter declined rapidly suggesting initial tumor killing functions in these specific immune aggregates. Despite initial reduction observed in milky spots, numbers of tumor cells however increased at fatty tissue stripes that border the omentum. This indicated proliferation at these locations, which corresponded to macroscopic observations of the omenta on day 21 after tumor cell injection. Omentectomy resulted in reduced intra-abdominal tumor load, which was completely attributable to the absence of the omentum, as tumor development did not differ on other sites. Even in the MRD group microscopic clusters of tumor cells located in the omentum eventually developed into macroscopic nodules. CONCLUSION: Since the ability of omental milky spots is, even in MRD, insufficient to prevent intra abdominal tumor outgrowth, omentectomy, which reduces tumor load, is recommended in surgical treatment of intra abdominal tumors that are prone to disseminate intraperitoneally. PMID- 16311733 TI - Growth inhibition of breast cancer cell lines overexpressing Her2/neu by a novel internalized fully human Fab antibody fragment. AB - The Her2/neu oncogene is overexpressed in various human cancers of epithelial origin and is associated with increased metastatic potential and poor prognosis. Blocking the Her2/neu signalling has been the focus of most therapeutic approaches. In this paper, the Her2/neu extracellular domain expressed in soluble form in yeast Pichia pastoris was used in order to isolate a fully human Fab fragment from a combinatorial Fab phage display library, derived from invaded lymph nodes of a breast cancer patient. The isolated fully human Fab63 binds specifically the native Her2/neu receptor and competes with Herceptin for binding to soluble Her2/neu receptor. In Her2/neu overexpressing cancer cells, Fab63 is rapidly internalized and has significant antiproliferative effects, where ligand independent mechanisms dominate signal induction. Moreover, in the presence of the ligand heregulin, growth inhibition was also detected by Fab63. The human Fab63 is a non-immunogenic agent with unique properties that can be applied in diagnosis and cancer therapy, with great potential for further manipulation towards the generation of an effective anticancer molecule. PMID- 16311734 TI - Clinical relevance of NK, NKT, and dendritic cell dose in patients receiving G CSF-mobilized peripheral blood allogeneic stem cell transplantation. AB - To analyze the relationship between the cellular composition of peripheral blood allografts and clinical outcome, we performed a prospective study in 45 adult patients who underwent allogeneic peripheral blood hematopoietic stem cell transplantation (HSCT) from a histocompatibility leukocyte antigen identical sibling donor for different hematological malignancies. The dose of CD34+, CD3+, CD4+, CD8+, and CD19+ lymphocytes, natural killer (NK) cells, natural killer T (NKT) cells, type 1 and type 2 dendritic cells (DC1 and DC2), as well as regulatory T (Treg) lymphocytes was analyzed. All patients were conditioned with busulphan and cyclophosphamide (BuCy2) +/- VP-16 and received a short course of methotrexate and cyclosporin-A as graft-versus-host disease (GVHD) prophylaxis. Acute GVHD (aGVHD) was present in 9 of 43 (21%) patients, and chronic GVHD (cGVHD) developed in 18 of 39 (46%) patients. There was a significantly higher incidence of aGVHD in patients receiving more than 6x10(6)/kg CD34+ cells. In univariate analysis, variables associated with better survival were as follows: a dose of less than 1.5x10(7)/kg NKT cells and less than 1.7x10(6)/kg DC2 for disease-free survival (DFS), and a dose of less than 3x10(7)/kg NK cells, less than 1.5x10(7)/kg NKT cells, less than 3x10(6)/kg DC1, and less than 1.7x10(6)/kg DC2 for overall survival (OS). In the Cox regression analysis, the dose of NKT cells was the only variable associated with better DFS, while the doses of NK, NKT, and CD34+ cells (less than 8x10(6)/kg) were associated with better OS. In conclusion, different circulating cell populations, other than CD34+ cells, are also of relevance in predicting the clinical outcome after allogeneic peripheral blood HSCT. PMID- 16311735 TI - Spontaneous tumor lysis syndrome in a patient with diffuse large B cell lymphoma and Richter syndrome. PMID- 16311736 TI - Complete remission and successful stem cell mobilization after treatment of refractory plasma cell leukemia with bortezomib. PMID- 16311737 TI - Rare association of severe hypoplasia of the abdominal aorta with imperforate anus, colonic atresia, and choledochal cyst. AB - Hypoplasia of the abdominal aorta (HAA) is a rare condition that causes marked hypertension. Although multiple etiologies have been postulated for HAA, congenital structural anomalies are rarely observed except in cases associated with some hereditary syndromes. The authors describe a neonatal case with HAA complicated by multiple anomalies including colonic atresia (CA), imperforate anus, choledochal cyst, facial cleft, and brain defects. This patient showed CA in the descending colon and caliber change in the transverse colon mimicking Hirschsprung disease, both of which were thought to be caused by vascular insult to the mesentery due to HAA. Although multiple surgical corrections were successfully performed, the hypertension was uncontrollable. PMID- 16311738 TI - Identification of tissue-specific vasculogenic cells originating from murine uterus. AB - Endometrium is a highly regenerative adult tissue that undergoes repeated degeneration and regeneration following menarche. Therefore, it is believed that endometrium contains stem and/or progenitor cells in order to compensate for the regeneration of tissue components. We report here that stem-like cells having vasculogenic potential are present in the uterus. Enzymatically extracted cells from murine uteri were characterized and fractionated into four subpopulations by flowcytometry; CD34(+)/45(-) (Ut-34), CD34(-)/45(-) (Ut-DN) and the remaining CD45(+) cell fractions (CD34(+)/45(+) and CD34(-)/45(+) cells). The Ut-34 and Ut DN fractions were mostly negative for putative endothelial cell (EC) markers, such as CD31, Flk-1, c-kit and VE-cadherin, although the Ut-DN fraction contained 2.8% CD31(+) cells. Ut-DN cells were further divided into CD31(+) and CD31(-) fractions. Three cell populations were obtained from green fluorescence protein (GFP) transgenic mice and were transplanted into injured wild-type mouse skeletal muscle. At 4 weeks after cell transplantation, donor-derived vascular smooth muscle and ECs were observed in the injured recipient muscle. A similar trend was observed in the Ut-34 group, but differentiation into vascular smooth muscle was predominant. In contrast, the Ut-DN/31(+) cell-transplanted group showed preferential differentiation into vascular ECs, thus suggesting that they were relatively committed preexisting ECs. These characteristics were also seen in vitro, in clonal cell cultures. Interestingly, donor derived Ut-DN/31(+), Ut DN/31(-) and Ut-34 cells could not be identified after bone marrow (BM) transplantation, thus confirming that they are not derived from BM. It therefore appeared that tissue-specific vasculogenic cells are present in the murine uterus and that they exhibit vascular formation, even in different tissue microenvironments. PMID- 16311739 TI - Evaluation of pediatric head trauma in the emergency department. PMID- 16311740 TI - Predictors of intracranial injuries in children after blunt head trauma. AB - This study was conducted to determine if clinical features can predict the risk of intracranial injury (ICI) in pediatric closed head trauma. We enrolled 3,806 children under 16 years consecutively referred for acute closed head trauma to the paediatric emergency room of five Italian children's hospitals. Relevant outcomes were death and diagnosis of ICI. Clinical symptoms and signs were evaluated as possible outcome predictors. Children were also classified into five groups according to their clinical presentation. The association of ICI with signs and symptoms and the appropriateness of the five-group classification in predicting the likelihood of ICI were evaluated by logistic regression analyses. ICI was diagnosed in 22 children; 2 of them died. The risk of fatal and nonfatal ICI was 0.5 and 5.2 per 1,000 children with closed head trauma respectively. Significant associations were found between ICI and loss of consciousness, prolonged headache, persistent drowsiness, abnormal mental status, focal neurological signs, signs of skull fracture in non-frontal areas and signs of basal skull fracture. The five-group classification of children allowed an excellent prediction in terms of likelihood of ICI (ROC area 0.972). CONCLUSIONS: Selection of children with closed head trauma based on different combinations of signs and symptoms allows for early identification of subjects at different risk for ICI. In patients with minor head injuries, the absence of loss of consciousness, drowsiness, amnesia, prolonged headache, clinical evidence of basal or non-frontal skull fracture identified 100% of children without lesions. Validation of our results with a larger sample of patients with ICI would be highly desirable. PMID- 16311741 TI - Prevalence and associated factors for temporomandibular disorders in a group of Mexican adolescents and youth adults. AB - The objective of the study was to determine the prevalence and associated factors for temporomandibular disorders (TMD) in a university sample of Campeche, Mexico. A cross-sectional study was carried out in 506 subjects aged 14-25 years. The subjects were requested to answer questionnaires concerning sociodemographic variables, history of stress, lifestyle, and anxiety. The Research Diagnostic Criteria for TMD (RDC/TMD) was used as TMD diagnostic system by four examiners capacitated and standardized. Data were analyzed using binary logistic regression in STATA. The results showed that 46.1% of the subjects exhibited some grade of TMD. Logistic regression analysis with TMD as the dependent variable identified sex (women odds ratio [OR]=1.7), bruxism (OR=1.5), anxiety (OR=1.6), unilateral chewing (OR=1.5), and an interaction between number of tooth loss and stress as the most significant associated variables, thus (1) the effect of having high levels of stress in the group of subjects without tooth loss (OR=1.2; 95% confidence interval [CI]=0.7-1.8) and (2) the effect of having high levels of stress in the group of subjects with at least one tooth lost (OR=2.4; 95% CI=1.01 5.9). The variables associated with diagnosis of pain were principally psychosocial (stress and anxiety), whereas for the non-pain diagnosis group, the variables were clinical, such as bruxism, chewing site preference, and restorations in mouth. We found associations among variables that were similar to findings in other studies, such as bruxism, tooth loss, stress, and anxiety. The final model explains that the effect of stress on TMD depends of the tooth loss, controlling for sex, bruxism, unilateral chewing, and anxiety. Finally, it can be concluded that the variables associated with pain and non-pain diagnosis were of distinct nature. PMID- 16311742 TI - Miniplate osteosynthesis of fractures of the edentulous mandible. AB - This study was performed to analyze treatment of fractures of the edentulous mandible and to discuss this method in relation to the mandibular height at the fracture site. Fifteen fracture sites in 11 patients with an edentulous mandible were retrospectively examined. These fractures were located: nine fractures in the mandibular body, three in the paramedian region, and three in the mandibular angle. Fractures in a mandible measuring more than 10 mm in the vertical height were treated with one miniplate. Fractures in an extremely atrophic mandible with 10 mm or less were treated using one or two miniplates, also using a modified Champy plate with 1.3 mm in thickness. A mandibular fracture with a height of 5 mm was treated with a combination of a microplate on the buccal side and a miniplate on the inferior border of the mandible with additional direct circumferential wiring. Oblique or splitting fractures were treated with direct circumferential wiring or a Herbert screw, at one fracture site each, respectively. Complications, including infection, fibrous union, nonunion and trismus, were not seen. In one patient, hypesthesia of the lower lip was, however, persistent 1 month after surgery. Miniplate osteosynthesis is the less invasive treatment, and it is suitable for fractures of the atrophic edentulous mandible, except for comminuted or defect fractures. To obtain stable fixation in severely atrophic mandibles, we need to consider the use of two miniplates or a combination with microplates. PMID- 16311743 TI - End-of-life care: is euthanasia the answer? PMID- 16311744 TI - Life-threatening subcutaneous hemorrhage following minor blunt trauma in an elderly patient taking ticlopidine and aspirin: a case report. AB - We report a rare case in which life-threatening subcutaneous hemorrhage following minor blunt trauma developed in an elderly patient taking ticlopidine and aspirin. An 82-year-old woman taking ticlopidine and aspirin tripped on a road. She noted only mild pain at the left lower abdomen. However, the hematoma gradually enlarged, and she developed hemorrhagic shock. Emergently performed contrast-enhanced abdominal computed tomography showed a large hematoma (20 x 10 x 7 cm), including contrast extravasation, which was located beneath the skin of the left lower abdominal quadrant and within the external and internal oblique muscles. Emergency pelvic arteriography showed massive contrast extravasation from the left superficial iliac circumflex artery. Transcatheter arterial embolization (TAE) was successfully performed, using gelatin sponge pledgets and micro-size stainless steel coils. After TAE, hemodynamics stabilized. The patient was discharged uneventfully on day 12. PMID- 16311745 TI - SNP array-based homozygosity mapping reveals MCPH1 deletion in family with autosomal recessive mental retardation and mild microcephaly. AB - Very little is known about the molecular basis of autosomal recessive MR (ARMR) because in developed countries, small family sizes preclude mapping and identification of the relevant gene defects. We therefore chose to investigate genetic causes of ARMR in large consanguineous Iranian families. This study reports on a family with six mentally retarded members. Array-based homozygosity mapping and high-resolution microarray-based comparative genomic hybridization (array CGH) revealed a deletion of approximately 150-200 kb, encompassing the promoter and the first six exons of the MCPH1 gene, one out of four genes that have been previously implicated in ARMR with microcephaly. Reexamination of affected individuals revealed a high proportion of prematurely condensed chromosomes, which is a hallmark of this condition, but in spite of the severity of the mutation, all patients showed only borderline to mild microcephaly. Therefore the phenotypic spectrum of MCPH1 mutations may be wider than previously assumed, with ARMR being the only consistent clinical finding. PMID- 16311748 TI - Rationale for a diagnostic chain in gestational breast tumor diagnosis. AB - Due to the physiological tissue alterations of the breast during pregnancy and lactation the diagnosis of a breast tumor by palpation, imaging and fine-needle biopsy is difficult and often unreliable. The purpose of our study was to review the value of imaging (mammography and ultrasonography) and biopsy techniques in order to work out a reliable protocol for evaluating gestational tumors. We performed a retrospective analysis of 25 patients with clinical diagnosis of gestational breast tumors during a 4-year period from 1995 to 1999. Specimens were obtained by core biopsy for histological examination. Invasive breast cancer was diagnosed in five patients, malignant lymphoma of the skin in one patient. The tumors in the remaining 19 patients proved benign. The overall accuracy was 0.91 for conventional mammography, 0.76 for conventional and panoramic ultrasound and 0.48 for color-coded sonography. Mammography as the most reliable imaging method should be used to evaluate all patients in whom ultrasonographic findings are suspicious for tumor. Radiation phobia should not prevent the performance of mammography under these circumstances. Nevertheless, all pre-operative methods of diagnosing gestational breast tumors other than core biopsy are unreliable. PMID- 16311749 TI - Emergency obstetric hysterectomy in Kuwait: a clinico pathological analysis. AB - The major objective of this study is to analyze the histological findings in emergency obstetric hysterectomy specimens and correlate them with the clinical diagnosis, epidemiological factors and number of tissue blocks examined. The records of all peripartum hysterectomy specimens examined between 1995 and 2001 in the Department of Pathology, Al-Sabah Hospital Kuwait were analyzed. A minimum of ten blocks from the cervix or lower uterine segment was arbitrarily regarded as adequate. Fifty-eight patients had peripartum hysterectomies (incidence of 0.07% of all deliveries) out of which 65% were aged between 30 and 40 years. The number of hysterectomy increased with parity up to five and then declined. The number of blocks examined varied from 2 to 53. Satisfactory pathology was found in 40 and absent in 18 cases. There were 33 adherent placentas. Amniotic fluid embolism (AFE) was found in seven patients. Adequate tissue blocks (>or=10) were significantly associated with positive pathological findings. P value <0.05. PMID- 16311750 TI - The high-grade endometrial sarcoma: a rare entity. AB - INTRODUCTION: With an estimated incidence of one to two per one million women, the endometrial stromal sarcoma (ESS) is a rare disease. It is subclassified into a high-grade and a prognostically better low-grade type. Evidence-based data for a standardized therapy is lacking. CASE REPORT: A 32-year-old obese nulligravida presented with persistent vaginal bleeding after the operation of an acute adnextorsion at another hospital. The repeat gynecological ultrasound examination showed a 5.3x5.3x3.6 cm vascularized, partially inhomogeneous mass in the uterus. A fractioned curettage yielded a differential diagnosis of malignant muellerian mixed tumor or a non-differentiated endometrial sarcoma. For completion of the operative treatment, laparotomy with hysterectomy, adnexectomy, and pelvine lymphonodectomy were performed. The final histological report described a 7 cm non-differentiated endometrial sarcoma with infiltration of the left ovary and 25 tumor-free lymph nodes. DISCUSSION: Standard therapy for resectable sarcoma is abdominal hysterectomy and bilateral adnexectomy. So far, there is little data from studies reporting radio- or chemotherapy treatment of small patient numbers in an adjuvant setting. CONCLUSION: The ESS is a very rare disease of the uterus. Due to missing clinical data, it remains a multidisciplinary therapeutic challenge requiring individual decisions. To receive more information on this rare disease, treatment should be performed according to international protocols. PMID- 16311751 TI - Comparison of a biopsychosocial therapy (BT) with a conventional biomedical therapy (MT) of subacute low back pain in the first episode of sick leave: a randomized controlled trial. AB - This randomized controlled clinical trial compares the effectiveness of a biopsychosocial treatment with a solely conventional biomedical therapy in patients with subacute low back pain using parameters for pain intensity, functional status, depressive dysfunction and work performance. Sixty-four patients with a first-time sick leave between 3 and 12 weeks due to low back pain were randomly assigned to either a conventional biomedical therapy (MT; n=33) group, or a biopsychosocial therapy (BT; n=31) group including a psychotherapeutic module; both in accordance with a standardized 3 weeks inpatient treatment. Pain intensity, functional back capacity, clinical parameters and depressive dysfunction revealed significant improvement in both treatment groups at end of 3 weeks therapy (T1). However, at 6 months (T2), analysis revealed significant better results for nearly all parameters in the BT group that showed further improvement from T1 to T2, whereas the values in the MT group deteriorated from T1 back to the baseline values. During the 2-year period after therapy, 10% in MT and 59% in BT required no further sick leave due to low back pain. The results of the study indicate that a psychotherapeutic element in the treatment of low back pain appears to positively influence pain, functional status and work performance when conducted at an early stage of chronification and helps in the achievement of a better outcome. PMID- 16311752 TI - Spinal loads after osteoporotic vertebral fractures treated by vertebroplasty or kyphoplasty. AB - Vertebroplasty and kyphoplasty are routine treatments for compression fractures of vertebral bodies. A wedge-shaped compression fracture shifts the centre of gravity of the upper body anteriorly and generally, this shift can be compensated in the spine and in the hips. However, it is still unclear how a wedge-shaped compression fracture of a vertebra increases forces in the trunk muscle and the intradiscal pressure in the adjacent discs. A nonlinear finite element model of the lumbar spine was used to estimate the force in the trunk muscle, the intradiscal pressure and the stresses in the endplates in the intact spine, and after vertebroplasty and kyphoplasty treatment. In this study, kyphoplasty represents a treatment with nearly full fracture reduction and vertebroplasty one without restoration of kyphotic angle although in reality kyphoplasty does not guarantee fracture reduction. If no compensation of upper body shift is assumed, the force in the erector spine increases by about 200% for the vertebroplasty but by only 55% for the kyphoplasty compared to the intact spine. Intradiscal pressure increases by about 60 and 20% for the vertebroplasty and kyphoplasty, respectively. In contrast, with shift compensation of the upper body, the increase in muscle force is much lower and increase in intradiscal pressure is only about 20 and 7.5% for the vertebroplasty and kyphoplasty, respectively. Augmentation of the vertebral body with bone cement has a much smaller effect on intradiscal pressure. The increase in that case is only about 2.4% for the intact as well as for the fractured vertebra. Moreover, the effect of upper body shift after a wedge-shaped vertebral body fracture on intradiscal pressure and thus on spinal load is much more pronounced than that of stiffness increase due to cement infiltration. Maximum von Mises stress in the endplates of all lumbar vertebrae is also higher after kyphoplasty and vertebroplasty. Cement augmentation has only a minor effect on endplate stresses in the unfractured vertebrae. The advantages of kyphoplasty found in this study will be apparent only if nearly full fracture reduction is achieved. Otherwise, differences between kyphoplasty and vertebroplasty become small or vanish. Our results suggest that vertebral body fractures in the adjacent vertebrae after vertebroplasty or kyphoplasty are not induced by the elevated stiffness of the treated vertebra, but instead the anterior shift of the upper body is the dominating factor. PMID- 16311753 TI - Os odontoideum with bipartite atlas and segmental instability: a case report. AB - We report on the case of a 15-year-old adolescent who presented with a transient paraplegia and hyposensibility of the upper extremities after sustaining a minor hyperflexion trauma to the cervical spine. Neuroimaging studies revealed atlantoaxial dislocation and ventral compression of the rostral spinal cord with increased cord signal at C1/C2 levels caused by an os odontoideum, as well as anterior and posterior arch defects of the atlas. The patient underwent closed reduction and posterior atlantoaxial fusion. We describe the association of an acquired instability secondary to an os odontoideum with an anteroposterior spondyloschisis of the atlas and its functional result after 12 months. The rare coincidence of both lesions indicates a multiple malformation of the upper cervical spine and supports the theory of an embryologic genesis of os odontoideum. PMID- 16311754 TI - Sagittal spinopelvic balance in normal children and adolescents. AB - The sagittal spinopelvic balance is poorly documented in normal pediatric subjects. The purpose of this study is to characterize the sagittal spinopelvic balance in the pediatric population and to evaluate the correlations between spinopelvic parameters. Seven parameters were evaluated from the lateral standing radiographs of 341 normal subjects aged 3-18 years old: thoracic kyphosis (TK), thoracic tilt (TT), lumbar lordosis (LL), lumbar tilt (LT), sacral slope (SS), pelvic tilt (PT) and pelvic incidence (PI). The mean values for the pelvic parameters were 49.1+/-11.0, 7.7+/-8.0 and 41.4+/-8.2 degrees for PI, PT and SS, respectively. The mean values for the spinal parameters were 48.0+/-11.7, 44.0+/ 10.9, -7.3+/-5.2 and -3.1+/-5.2 degrees for LL, TK, LT and TT, respectively. The spinopelvic parameters were different from those reported in normal adults, but the correlations between the parameters were similar. PI was significantly related to SS and PT. Significant correlations were found between the parameters of adjacent anatomical regions. Pelvic morphology (PI) regulates sagittal sacro pelvic orientation (SS and PT). Sacral orientation (SS) is correlated with the shape (LL) and orientation (LT) of the lumbar spine. Adjacent anatomical regions of the spine and pelvis are interdependent, and their relationships result in a stable and compensated posture, presumably to minimize energy expenditure. Results from this study could be used as an aid for the planning of surgery in pediatric patients with spinal deformity in order to restore a relatively normal sagittal spinopelvic balance. PMID- 16311755 TI - Relationship between Northwick Park neck pain questionnaire and cervical spine MR imaging findings. AB - The study was aimed at determining the association between the self-report of pain and disability by means of Northwick neck pain questionnaire (NPQ) and cervical spine MR imaging findings. A random sample of 251 patients, 132 men and 119 women aged 43+/-13 years, submitted with neck pain were investigated. Patients with previous discitis, surgery, neoplasm or hospitalized for cervical spine trauma were excluded. All patients completed the NPQ and were studied with sagittal gradient-echo T1 and turbo spin-echo T2, axial gradient-echo T2* and heavily T2 weighted MR myelographic weighted images. MR images of the two most affected disc levels were read, offering an MR imaging score from 0 to 30. There was no statistically significant correlation between NPQ and MR imaging scores. From the NPQ items, only difficulty in sleeping and numbness were related to the MR imaging score. Disc extrusion was the only MR finding almost significantly associated with NPQ (P=0.054). Neck injury did not increase NPQ scores. In patients with neck pain, NPQ scores do not correlate with MR imaging findings. NPQ and cervical spine MR imaging show different facets of the multidimensional complex of neck pain. PMID- 16311756 TI - Ordering of p-n-alkoxybenzoic acids at phase transition temperatures: a comparative computational analysis. AB - A comparative analysis of molecular ordering of nematogenic p-n-alkoxybenzoic acids has been carried out with respect to translatory and orientational motions for the acids with seven (7OBAC), eight (8OBAC), nine (9OBAC) and 10 (10OBAC) carbon atoms in the alkyl chain. The CNDO/2 method has been used to compute the net atomic charge and dipole moment components at each atomic center. Modified Rayleigh-Schrodinger perturbation theory with multicentered-multipole expansion method has been used to evaluate long-range intermolecular interactions while a '6-exp' potential function has been assumed for short-range interactions. The total interaction-energy values obtained by these computations were used to calculate the probability of each configuration at the phase-transition temperature using the Maxwell-Boltzmann formula. The flexibility of various configurations has been studied in terms of variation of probability due to small departures from the most probable configuration. A comparative picture of molecular parameters like total energy, binding energy and total dipole moment has been given. An attempt has been made to explain the nematogenicity of these acids in terms of their relative order with the molecular parameter introduced in this paper. PMID- 16311757 TI - Visual rating of medial temporal lobe metabolism in mild cognitive impairment and Alzheimer's disease using FDG-PET. AB - PURPOSE: This study was designed to examine the utility of visual inspection of medial temporal lobe (MTL) metabolism in the diagnosis of mild cognitive impairment (MCI) and Alzheimer's disease (AD) using FDG-PET scans. METHODS: Seventy-five subjects [27 normal controls (NL), 26 MCI, and 22 AD] with FDG-PET and MRI scans were included in this study. We developed a four-point visual rating scale to evaluate the presence and severity of MTL hypometabolism on FDG PET scans. The visual MTL ratings were compared with quantitative glucose metabolic rate (MR(glc)) data extracted using regions of interest (ROIs) from the MRI-coregistered PET scans of all subjects. A standard rating evaluation of neocortical hypometabolism was also completed. Logistic regressions were used to determine and compare the diagnostic accuracy of the MTL and cortical ratings. RESULTS: For both MTL and cortical ratings, high intra- and inter-rater reliabilities were found (p values <0.001). The MTL rating was highly correlated with and yielded a diagnostic accuracy equivalent to the ROI MR(glc) measures (p values <0.001). The combination of MTL and cortical ratings significantly improved the diagnostic accuracy over the cortical rating alone, with 100% of AD, 77% of MCI, and 85% of NL cases being correctly identified. CONCLUSION: This study shows that the visual rating of MTL hypometabolism on PET is reliable, yields a diagnostic accuracy equal to the quantitative ROI measures, and is clinically useful and more sensitive than cortical ratings for patients with MCI. We suggest this method be further evaluated for its potential in the early diagnosis of AD. PMID- 16311758 TI - Analysis of a non-canonical NtcA-dependent promoter in Synechococcus elongatus and its regulation by NtcA and PII. AB - In this communication, we present a genetic analysis of the glnN promoter region of Synechococcus elongatus PCC 7942. luxAB reporter fusions were used to characterize the glnN promoter by deletion and site-directed mutational analysis. Reporter gene expression analysis was performed in S. elongatus wild-type and mutant strains to reveal the role of the global nitrogen responsive transcription factor NtcA and of the P(II) signalling protein on regulation of glnN gene expression. A non-canonical NtcA-binding motif is responsible for NtcA-dependent, nitrogen-responsive regulation of glnN. The P(II) signalling protein has opposing effects on NtcA-dependent glnN expression. Under conditions of nitrate-growth, it depresses expression, whereas under conditions of combined nitrogen starvation, it is required for full induction. Furthermore, sequences upstream of the NtcA binding site have repressive effect on glnN promoter activity. PMID- 16311759 TI - Analysing the outer membrane subproteome of Methylococcus capsulatus (Bath) using proteomics and novel biocomputing tools. AB - High-resolution two-dimensional gel electrophoresis and mass spectrometry has been used to identify the outer membrane (OM) subproteome of the Gram-negative bacterium Methylococcus capsulatus (Bath). Twenty-eight unique polypeptide sequences were identified from protein samples enriched in OMs. Only six of these polypeptides had previously been identified. The predictions from novel bioinformatic methods predicting beta-barrel outer membrane proteins (OMPs) and OM lipoproteins were compared to proteins identified experimentally. BOMP ( http://www.bioinfo.no/tools/bomp ) predicted 43 beta-barrel OMPs (1.45%) from the 2,959 annotated open reading frames. This was a lower percentage than predicted from other Gram-negative proteomes (1.8-3%). More than half of the predicted BOMPs in M. capsulatus were annotated as (conserved) hypothetical proteins with significant similarity to very few sequences in Swiss-Prot or TrEMBL. The experimental data and the computer predictions indicated that the protein composition of the M. capsulatus OM subproteome was different from that of other Gram-negative bacteria studied in a similar manner. A new program, Lipo, was developed that can analyse entire predicted proteomes and give a list of recognised lipoproteins categorised according to their lipo-box similarity to known Gram-negative lipoproteins ( http://www.bioinfo.no/tools/lipo ). This report is the first using a proteomics and bioinformatics approach to identify the OM subproteome of an obligate methanotroph. PMID- 16311760 TI - Ganglion cysts of the proximal tibiofibular joint review of literature with three case reports. AB - Proximal tibiofibular ganglion is a rare disorder. It may settle down in the subcutaneous tissue or may develop along the peroneal muscles and nerve. Common clinical findings are various sizes of mass, pain and hypoesthesis due to compression neuropathy. We report three cases of proximal tibiofibular ganglion and review the literature about the diagnostic tools, recurrence rates and treatment modalities. PMID- 16311761 TI - Trans-sacral fixation for failed posterior fixation of the pelvic ring. AB - INTRODUCTION: In the treatment of certain pelvic ring pathologies (non-unions and failure of ilio-sacral screw fixation) trans-sacral fixation (i.e. fixation from iliac wing to the other traversing the body of S1) may be necessary. The purpose of our study was to describe our early experience and describe the surgical technique. MATERIALS AND METHODS: Seven cases of trans-sacral fixation were identified with a mean patient age of 49-years-old with four females and three males. The diagnosis at the time of trans-sacral fixation was failure of posterior ring fixation in four (three of which had vertical sacral fractures), mal-position of iliosacral (IS) screw in one, failure of fusion of sacroiliac (SI) joint in two. All these cases were augmented with a 4.5 mm reconstruction plate placed in tension posteriorly. RESULTS: At average follow up of 39 months (range 24-75), all patients achieved union. There were no neurological or vascular complications. Two patients required reoperations prior to union. Our current use for this technique is with a 8.0 mm screw (16 mm thread). CONCLUSIONS: Initial experience with trans-sacral fixation has proven to be very effective technique to solve the most difficult problems in posterior pelvic ring fixation. We reserve its use to the following indications: nonunion/malunion of the pelvic ring, and sacral fractures. PMID- 16311763 TI - Gene expression in skeletal muscle of coronary artery disease patients after concentric and eccentric endurance training. AB - Low-intensity concentric (CET) and eccentric (EET) endurance-type training induce specific structural adaptations in skeletal muscle. We evaluated to which extent steady-state adaptations in transcript levels are involved in the compensatory alterations of muscle mitochondria and myofibrils with CET versus EET at a matched metabolic exercise intensity of medicated, stable coronary patients (CAD). Biopsies were obtained from vastus lateralis muscle before and after 8 weeks of CET (n=6) or EET (n=6). Transcript levels for factors involved in mitochondrial biogenesis (PGC-1alpha, Tfam), mitochondrial function (COX-1, COX 4), control of contractile phenotype (MyHC I, IIa, IIx) as well as mechanical stress marker (IGF-I) were quantified using an reverse-transcriptase polymerase chain reaction approach. After 8 weeks of EET, a reduction of the COX-4 mRNA level by 41% and a tendency for a drop in Tfam transcript concentration (-33%, P=0.06) was noted. This down-regulation corresponded to a drop in total mitochondrial volume density. MyHC-IIa transcript levels were specifically decreased after EET, and MyHC-I mRNA showed a trend towards a reduction (P=0.08). Total fiber cross-sectional area was not altered. After CET and EET, the IGF-I mRNA level was significantly increased. The PGC-1alpha significantly correlated with Tfam, and both PGC-1alpha and Tfam significantly correlated with COX-1 and COX-4 mRNAs. Post-hoc analysis identified significant interactions between the concurrent medication and muscular transcript levels as well as fiber size. Our findings support the concept that specific transcriptional adaptations mediate the divergent mitochondrial response of muscle cells to endurance training under different load condition and indicate a mismatch of processes related to muscle hypertrophy in medicated CAD patients. PMID- 16311764 TI - Linear decrease in .VO2max and performance with increasing altitude in endurance athletes. AB - It has been hypothesized that one reason for decreased .VO(2max) in hypoxia could be the lower maximal exercise intensity achieved in incremental, time or distance trial tests. We hypothesized that (1).VO(2max) would be decreased at altitude even when exercising at the same absolute maximal exercise intensity as at sea level and; (2) the decline in .VO(2max) in endurance-trained athletes (ETA) would be linear across the range from sea level through moderate altitudes. Eight ETA performed combined .VO(2max) and performance tests running to exhaustion at the same speed in a randomized double blind fashion at simulated altitudes of 300, 800, 1,300, 1,800, 2,300 and 2,800 m above sea level using a hypobaric chamber. Douglas bag system was used for respiratory measurements and pulse oximetry was used to estimate arterial O(2) saturation. .VO(2max) declined linearly from 66+/ 1.6 ml kg(-1) min(-1) at 300 m to 55+/-1.6 ml kg(-1) min(-1) at 2,800 m corresponding to a 6.3% decrease per 1,000 m increasing altitude (range 4.6 7.5%). Time to exhaustion (performance) at a constant velocity associated with 107% of sea level .VO(2max) decreased with 14.5% (P<0.001) per 1,000 m altitude between 300 and 2,800 m. Both .VO(2max) and performance decreased from 300 to 800 m (P<0.01; P<0.05). Arterial haemoglobin oxygen saturation at test cessation (SpO(2min)) declined from 89.0+/-2.9% at 300 m to 76.5+/-4.0% at 2,800 m (P=0.001). This study report that in ETA during acute exposure to altitude both performance and .VO(2max) decline from 300 to 800 m above sea level and continued to decrease linearly to 2,800 m. PMID- 16311765 TI - Kinematic cues in perceptual weight judgement and their origins in box lifting. AB - When accepting a parcel from another person, we are able to use information about that person's movement to estimate in advance the weight of the parcel, that is, to judge its weight from observed action. Perceptual weight judgment provides a powerful method to study our interpretation of other people's actions, but it is not known what sources of information are used in judging weight. We have manipulated full form videos to obtain precise control of the perceived kinematics of a box lifting action, and use this technique to explore the kinematic cues that affect weight judgment. We find that observers rely most on the duration of the lifting movement to judge weight, and make less use of the durations of the grasp phase, when the box is first gripped, or the place phase, when the box is put down. These findings can be compared to the kinematics of natural box lifting behaviour, where we find that the duration of the grasp component is the best predictor of true box weight. The lack of accord between the optimal cues predicted by the natural behaviour and the cues actually used in the perceptual task has implications for our understanding of action observation in terms of a motor simulation. The differences between perceptual and motor behaviour are evidence against a strong version of the motor simulation hypothesis. PMID- 16311766 TI - Effects of lateral retinacular release on the lateral stability of the patella. AB - The objective of this cadaveric study was to evaluate quantitatively the effects of lateral retinacular release on the lateral stability of the patella. A materials testing machine was used to displace the patella of seven cadaveric specimens 10 mm laterally while measuring the required force, with 175 N quadriceps tension. The patella was connected via a ball-bearing patellar mounting 10 mm deep to the anterior surface to allow rotations. Patellar force- displacement behaviour was tested from 0 degrees to 60 degrees knee flexion. At 0 degrees , 10 degrees and 20 degrees flexion the mean force required to displace the patella 10 mm laterally was reduced significantly due to lateral retinacular release, by 16-19%. The average force required to displace the patella was also reduced for larger flexion angles, although this was not statistically significant. These findings suggest that lateral retinacular release may not be appropriate in treatment of patellar lateral instability. PMID- 16311767 TI - MRI-negative bucket-handle tears of the lateral meniscus in athletes: a case series. AB - Magnetic resonance imaging (MRI) is the most widely used non-invasive test for assessing intra-articular injuries of the knee. It has been suggested that a negative MRI can be useful in avoiding the need for diagnostic arthroscopy in cases where clinical examination is equivocal. However, the sensitivity and specificity of MRI is not 100%, particularly for tears of the lateral meniscus. Furthermore, a false negative MRI scan may result in premature return to play in athletes, resulting in increased risk of further damage to a torn meniscus. To illustrate this issue, we present a case series of eight elite athletes who all presented with mechanical knee symptoms and where MRI scans revealed no significant intra-articular pathology. Five of the athletes were allowed to return to sport on the basis of a negative MRI. All patients were subsequently found to have a bucket-handle tear of their lateral meniscus at arthroscopy. Two independent, experienced musculoskeletal radiologists were asked to review the MRI films without being given any clinical history of the cases. Although injuries to the popliteus tendon were noted in two of the athletes who had sustained a recent acute injury to their knee, no meniscal tears were identified. Thus arthroscopy remains the gold standard for the assessment and management of high-demand patients with a clinical suspicion of meniscal pathology. PMID- 16311768 TI - Brainstem gliomas. PMID- 16311769 TI - Influence of estrus status on urinary chemical parameters related to urolithiasis. AB - The present study examines the urinary chemical parameters related to urolithiasis in healthy female volunteers during premenopause and menopause, and discusses the role of menopause in stone formation. We investigated 24-h urine parameters associated with urinary stones and focused upon estrus status. Participants comprised 30 healthy women, 15 childless, premenopausal women and 15 menopausal women without a history of urolithiasis. Our results showed that menopausal women have lower citrate and higher calcium excretion, which might enhance calcium stone crystallization. We propose that the estrus status of female patients should be considered when evaluating metabolic abnormalities. PMID- 16311770 TI - An animal model of calcium oxalate urolithiasis based on a cyclooxygenase 2 selective inhibitor. AB - Our aim was to develop a stone-forming animal model involving renal tubular injury using a cyclooxygenase 2 selective inhibitor. Male Sprague-Dawley rats fed chow containing 3% sodium oxalate with or without 100 mg/kg celecoxib were compared to animals fed normal chow. Rats were killed after 2 or 4 weeks and the kidneys were harvested for morphological examination. Collections of 24-h urine were made before kidney harvest. After 2 weeks only a few crystals were observed in rats that received oxalate and celecoxib, but after 4 weeks more crystals were observed at the renal papilla than in rats that received only oxalate. Few crystals were found in rats fed normal chow with or without celecoxib. The urinary activities of gamma-glutamyl transpeptidase (GGT) were increased by celecoxib administration whereas creatinine clearance rates were unchanged. In rats fed oxalate, urinary oxalate excretion increased, but calcium excretion decreased. This model using a cyclooxygenase 2 selective inhibitor is a useful stone forming animal model involving mild renal tubular injury together with mild hyperoxaluria. PMID- 16311771 TI - The WHO classification of pituitary tumours: a combined neurosurgical and neuropathological view. PMID- 16311772 TI - Transition of areas of eosinophilic neurons and reactive astrocytes to delayed cortical infarcts after transient unilateral forebrain ischemia in Mongolian gerbils. AB - The fate of postischemic tissues containing eosinophilic neurons (ENs), whether they remain viable or evolve into infarction, is largely unknown. We analyzed the time profile and distribution of ENs, reactive astrocytes (RAs), and infarction after transient cerebral ischemia. Unilateral forebrain ischemia was induced in Mongolian gerbils by two 10-min unilateral common carotid artery occlusions with a 5-h interval, and the brains at 24 h, 4 days, and 2, 4, and 16 weeks were prepared for morphometric analysis. Intra-ischemic laser Doppler flowmetry revealed significant ischemia, deeper in the anterior cortex, during carotid occlusion. Here, ENs appeared in the middle and deep layers at 24 h postischemia, and EN areas had extended to all cortical layers by 4 days. Large areas of high EN density turned into infarcts between 4 days and 4 weeks. In the posterior cortex, middle and deep cortical layers evolved low EN density areas without subsequent transformation into infarcts. RAs were consistently observed in areas with ENs, and RA areas with high EN density were largely transformed into infarcts between 4 days and 4 weeks postischemia. Areas of high, but not low, EN density were slowly transformed into infarcts after transient cerebral ischemia. Delayed astrocytic death took place in the RA areas with high EN density. In conclusion, density of ENs is an important indicator of delayed astrocytic death and infarction in postischemic tissue. PMID- 16311773 TI - Harmonic scalpel tonsillectomy: a prospective study. AB - The harmonic scalpel has been in use for tonsillectomy for the last 5-6 years in western Europe and North America. Although some studies have found this technique to be superior to other conventional methods, its use is still not very popular. In this single-blinded prospective study, the intraoperative events and postoperative morbidity after the use of harmonic scalpel in tonsillectomy (HST) was evaluated in 180 cases in two hospitals and compared with conventional steel tonsillectomy (CST) and hemostasis secured by bipolar diathermy or ligatures in 100 cases. Both bipolar diathermy and ligatures were used to control the intraoperative bleeding in all cases of CST and some cases (n =9) of HST. The study was done in two hospitals. Patients were randomized irrespectively of their age, sex, past history or indication for surgery. The total number of patients operated on was 180 for HST and 100 for CST. The surgical duration, intraoperative blood loss and postoperative pain were compared between the patients who only had tonsillectomy done in either group (n =120 in HST and n = 70 in CST). The mean operative time in the HST group was not longer than the CST group, but the intraoperative blood loss was significantly less in the HST group. Postoperative pain was present in all patients in the HST group, but to a lesser extent than in the CST group. There was no major postoperative hemorrhage in the HST group that required surgical attention. HST has the advantage over CST when secondary hemorrhage after tonsillectomy is considered. Thus, following the results of the National Prospective Tonsillectomy Audit (NPTA), it may be safe to say that HST is superior to most other conventional methods in reducing secondary hemorrhage. The use of disposable blades in CST certainly reduces the risk of the transmission of Cruetzfield-Jacob disease (CJD). PMID- 16311774 TI - Validation of laser Doppler fluxmetry as a method of assessing neo-angiogenesis in laryngeal tumours. AB - Our intention was to validate the hypothesis that laser Doppler fluxmetry would give a valid representation of underlying tissue blood flow and would provide a reliable tool for measuring blood flow in human laryngeal cancers. This was an experimental prospective study. We performed laser Doppler fluxmetry on 20 patients with laryngeal cancer and compared the values obtained through this method with manual counting of microvessel density from biopsies, the accepted gold standard. We demonstrated good inter- and intraobserver variability in our estimation of microvessel density. We then compared microvessel density with laser Doppler fluxmetry, and using Spearman's test to assess nonparametric correlation, we obtained a correlation coefficient of 0.4 (P value: 0.15). To our knowledge, this study is the first of its kind to demonstrate a positive trend between angiogenesis measured conventionally when compared with laser Doppler fluxmetry. PMID- 16311801 TI - Transfer hydrogenation using recyclable polymer-supported formate (PSF): efficient and chemoselective reduction of nitroarenes. AB - Nitroarenes can be reduced in high yields to the corresponding anilines by transfer hydrogenation using a stable H-donor, polymer-supported formate (PSF) in combination with palladium acetate (catalytic). The reactions occur at 100-120 degrees C in dimethyl-formamide and the PSF can be recycled for at least three runs. The procedure is chemoselective for nitro group; ester, ketone, aldehyde, and halide substituents on aromatic ring remain unaffected. PMID- 16311802 TI - Polymer resin library and the discovery of highly efficient polymer supported reagents and scavengers. AB - This paper is focused on the preparation of a library of highly efficient, tailor made, macroporous resins as supports for reagents and scavengers. From this library a macroporous resin, prepared with 40% cross-linking and with a porogen level of 300% in toluene gave a material superior to those presently available, and was shown to be highly efficient in the preparation of novel highly reactive polymer supported reagents, including scavengers and oxidants. PMID- 16311803 TI - Cinchona alkaloid-based polymer-bound phase-transfer catalysts: efficient enantioselective alkylation of benzophenone imine of glycine esters. AB - Cross-linked polystyrene-bound and poly(ethylene glycol)-bound phase-transfer catalysts as well as homopolymers of cinchona alkaloid derivatives have been synthesised. Both soluble and insoluble polymers have been investigated. The enantioselective alkylation of N-diphenyl methylene glycine t-butyl ester has been successfully carried out in heterogeneous and homogeneous systems. High enantioselectivities (up to 96%) have been obtained. The polymer-bound catalysts have been easily recovered and conditions for efficient recycling have been studied. PMID- 16311804 TI - Rapid deprotection of N-Boc amines by TFA combined with freebase generation using basic ion-exchange resins. AB - The deprotection of N-Boc amines was rapidly accomplished using 5 equivalents of TFA in methylene chloride in a focused microwave instrument with irradiation at 60 degrees C for 30 min. The freebase amines are then obtained by scavenging the crude reaction mixture with the basic Amberlyst A-21 ion-exchange resin. This procedure is suitable for the parallel deprotection of N-Boc amines. PMID- 16311805 TI - Alternative method of Boc-removal from sulfamide using silica-phenyl sulfonic acid in conjunction with microwave heating. AB - A general method was used to prepare an array of unsymmetric sulfamides. This was accomplished by the stepwise addition of CSI to tert-butanol followed by the addition of amines. To increase diversity, nitrogen group of Boc-sulfamides was alkylated with alcohols using Mitsunobu reaction and Boc-group was removed using Si-TsOH. Microwave heating was used in all the steps. The final sulfamides were released from Si-TsOH using NH3 in MeOH. PMID- 16311806 TI - A convenient method to remove ruthenium byproducts from olefin metathesis reactions using polymer-bound triphenylphosphine oxide (TPPO). AB - Ruthenium byproducts from ring-closing metathesis reactions can be removed by refluxing the crude reaction mixture with resin-bound triphenylphosphine oxide (TPPO) in toluene or by stirring with dimethyl sulfoxide (DMSO) and silica gel at room temperature. Residual levels of ruthenium can be achieved that are as low as 0.04 microg per 5 mg of product when a combination of TPPO, DMSO, and silica gel is used. The polymer-bound TPPO retained its efficiency after being recycled six times. PMID- 16311807 TI - High-loading polyglycerol supported reagents for Mitsunobu- and acylation reactions and other useful polyglycerol derivatives. AB - In this paper we present soluble dendritic polyglycerol (PG) supported reagents PG-DEAD, PG-PPh3, and PG-DCC as well as scavengers PG-carbonate, PG-carbazate, and PG-amine, which all have been synthesized in high overall conversions and yields using simple purification techniques. The supported reagents have been used simultaneously in Mitsunobu and acylation reactions. All polymeric reagents and scavengers can be removed by simple precipitation/filtration protocols to give chromatography-free products of high purity. In the course of the syntheses of the polymeric reagents three intermediates turned out to be precious polyglycerol derivatives: a mixed carbonate as an electrophilic derivative, polyglyceryl carbazate as a scavenger for carbonyl compounds, as well as polyglycerylamines as amino analogues of polyglycerol. PMID- 16311808 TI - Knoevenagel reaction in water catalyzed by amine supported on silica gel. AB - An environmentally benign and sustainable Knoevenagel reaction of aldehyde with ethyl cyanoacetate has been achieved at ambient temperature in water employing 3 aminopropylated silica gel (NAP) as a catalyst. Wide applicability of the reaction is illustrated by the results that not only arylaldehydes of both electronic characters but also aliphatic aldehydes afforded the products. The reaction condition was so mild that aldehydes having acid- or base-sensitive substituents provided substituted alpha-cyano-alpha, beta-unsaturated esters. The catalyst has been efficiently recycled more than five times without any pre treatment. Catalyst loading was successfully reduced to 0.0029 mmol% (TON = up to 9,226). This protocol was also applicable to the Knoevenagel reaction of malononitrile in good yields in water. PMID- 16311809 TI - Polymer-supported and polymeric chiral guanidines: preparation and application to the asymmetric Michael reaction of iminoacetate with methyl vinyl ketone. AB - Polymer-supported and polymeric chiral guanidines carrying an imidazolidine skeleton are designed as polymer-based base catalysts. Thus, (R)-2-[(S)-1 hydroxymethyl-2-phenylethylimino]-4-phenylimidazolidine was newly prepared from (R)-phenylglycine as the key functional unit of these guanidines. Application of the polymer-based chiral guanidines to the asymmetric Michael reaction of t-butyl diphenyliminoacetate with MVK led to expected asymmetric induction in the Michael adduct with moderate enantioselectivity in the use of the latter polymeric chiral guanidines. PMID- 16311810 TI - 2-Pyridinealdoxime, a new ligand for a Pd-precatalyst: application in solid-phase assisted Suzuki-Miyaura reaction. AB - The preparation of a new palladium(II) complex with a 2-pyridinealdoxime ligand and its use as a Pd(0) precatalyst in the cross-coupling Suzuki-Miyaura reaction is described. Several concepts for the immobilization of this catalytic system are presented and compared in order to develop an efficient catalytic tool for high-throughput synthesis. PMID- 16311811 TI - A practical synthesis of a high-loading solid-supported IBX amide for the oxidation of alcohols. AB - A straightforward three-step synthesis of a solid-supported IBX amide resin was achieved using inexpensive and commercially available 2-iodobenzoic acid chloride and Merrifield resin. A high apparent loading of 0.63 mmol g(-1) was obtained. Oxidation of a range of alcohols to the corresponding carbonyl compounds proved very straightforward using 1.2 equiv of the resin. Recycling of the resin was also possible with minimal loss of activity after two reoxidations. PMID- 16311812 TI - Fluorous reagents and scavengers versus solid-supported reagents and scavengers, a reaction rate and kinetic comparison. AB - Reactions using fluorous reagents and scavengers are compared side-by-side with their solid-supported counterparts. Fluorous triphenylphosphine is used in the bromination reaction of alcohols, fluorous thiol is used as an electrophile scavenger for alpha-bromoketones, fluorous isatoic anhydride is used as a nucleophile scavenger for primary and secondary amines. Reactions involving fluorous reagents and scavengers occur in homogeneous media with solution-phase reaction kinetics. Reactions with solid-supported reagents and scavengers occur in a heterogeneous media, and the reaction kinetics are greatly affected by the nature of the solid-support and reaction environment. Significantly larger amounts of reagents and more time are usually required to complete the solid supported reaction. PMID- 16311813 TI - Parallel and automated library synthesis of 2-long alkyl chain benzoazoles and azole[4,5-b]pyridines under microwave irradiation. AB - A versatile route to 40-membered library of 2-long alkyl chain substituted benzoazoles (1 and 2) and azole[4,5-b]pyridines (3 and 4) via microwave-assisted combinatorial synthesis was developed. The reactions were carried out in both monomode and multimode microwave oven. With the latter, all reactions were performed in high-throughput experimental settings consisting of an 8 x 5 combinatorial library designed to synthesize 40 compounds. Each step, from the addition of reagents to the recovery of final products, was automated. The microwave-assisted N-long chain alkylation reactions of 2-alkyl-1H-benzimidazole (1) and 2-alkyl-1H-benzimidazole[4,5-b] pyridines (3) were also studied. PMID- 16311814 TI - Multi-space classification for predicting GPCR-ligands. AB - A classification of molecules depends on the descriptor set which is used to represent the compounds, and each descriptor could be regarded as one perception of a molecule. In this study we show that a combination of several classifiers that are grounded on separate descriptor sets can be superior to a single classifier that was built using all available descriptors. The task of predicting ligands of G-protein coupled receptors (GPCR) served as an example application. The perceptron, multilayer neural networks, and radial basis function (RBF) networks were employed for prediction. We developed classifiers with and without descriptor selection. Prediction accuracy was assessed by the area under the receiver operating characteristic (ROC) curve. In the case with descriptor selection both the selection and the rank order of the descriptors depended on the type and topology of the neural networks. We demonstrate that the overall prediction accuracy of the system can be improved by joining neural network classifiers of different type and topology using a "jury network" that is trained to evaluate the predictions from the individual classifiers. Seventy-one percent correct prediction of GPCR ligands was obtained. PMID- 16311815 TI - Molecular diversity of rat brain proteins as revealed by proteomic analysis. AB - Multiple protein expression forms (MPEFs) presenting splicing forms or co- and posttranslation modifications, account for the vast diversity, the myriad of gene products and clearly indicate problems which proteomics research is facing. In the present study, we generated a rat brain map representing MPEFs by the use of an analytical method based on two-dimensional electrophoresis combined with mass spectrometry. Forty-nine individual proteins were selected that showed more than two spots, resulting altogether into a total number of 357 expression forms. Some proteins showed large MPEFs numbers as e.g. tubulin alpha-1 chain (24 spots), ATP synthase alpha-chain (28), beta chain (17) or septin 7 (13). The molecular diversity observed in this map clearly shows that immunochemical or even protein chemical results from expressional studies have to be interpreted with caution, in particular if one dimensional electrophoretic or western blot techniques are applied and MPEFs are poorly resolved. PMID- 16311819 TI - Methodological scheme for designing the monitoring of genetically modified crops at the regional scale. AB - According to EC regulations the deliberate release of genetically modified (GM) crops into the agro-environment needs to be accompanied by environmental monitoring to detect potential adverse effects, e.g. unacceptable levels of gene flow from GM to non-GM crops, or adverse effects on single species or species groups thus reducing biodiversity. There is, however, considerable scientific and public debate on how GM crops should be monitored with sufficient accuracy, discussing questions of potential adverse effects, agro-environmental variables or indicators to be monitored and respective detection methods; Another basic component, the appropriate number and location of monitoring sites, is hardly considered. Currently, no consistent GM crop monitoring approach combines these components systematically. This study focuses on and integrates spatial agro environmental aspects at a landscape level in order to design monitoring networks. Based on examples of environmental variables associated with the cropping of Bt-Maize (Zea maize L.), herbicide-tolerant (HT) winter oilseed rape (Brassica napus L.), HT sugar beet (Beta vulgaris L.), and starch-modified potato (Solanum tuberosum L.), we develop a transferable framework and assessment scheme that comprises anticipated adverse environmental effects, variables to be measured and monitoring methods. These we integrate with a rule-based GIS (geographic information system) analysis, applying widely available spatial area and point information from existing environmental networks. This is used to develop scenarios with optimised regional GM crop monitoring networks. PMID- 16311820 TI - The new Danish stream monitoring programme (NOVANA)--preparing monitoring activities for the Water Framework Directive era. AB - Denmark has a long tradition of monitoring the aquatic environment. Previous monitoring has mainly focused on loss of nutrients and subsequent impacts on the biological structure in lakes and coastal areas. However, as part of the third Action Plan for the Aquatic Environment more emphasis has been put on stream ecology. The present paper describes background, strategy and content of the new NOVANA stream programme, which will run for the period 2004-2009. The new programme will encompass more than 800 stations covering all stream types in Denmark and monitoring will include three biological quality elements (macrophytes, macroinvertebrates and fish) as well as physico-chemical features and hydromorphological elements. In addition, the new programme integrates monitoring of elements both in the stream itself and in the riparian zone. Compliance with important European Commission Directives such as the Water Framework Directive and the Habitat Directive is discussed. PMID- 16311821 TI - Source apportionment of groundwater pollution around landfill site in Nagpur, India. AB - The present work attempts statistical analysis of groundwater quality near a Landfill site in Nagpur, India. The objective of the present work is to figure out the impact of different factors on the quality of groundwater in the study area. Statistical analysis of the data has been attempted by applying Factor Analysis concept. The analysis brings out the effect of five different factors governing the groundwater quality in the study area. Based on the contribution of the different parameters present in the extracted factors, the latter are linked to the geological setting, the leaching from the host rock, leachate of heavy metals from the landfill as well as the bacterial contamination from landfill site and other anthropogenic activities. The analysis brings out the vulnerability of the unconfined aquifer to contamination. PMID- 16311822 TI - Environmental quality of urban parks and open spaces in Hong Kong. AB - It is widely purported that urban parks and open spaces can improve the urban environment by cleansing the air and reducing the noise. However, such environmental functions can be constrained by the size and the location of urban parks in very dense cities. This study examines the air and acoustic environment and heavy metal contamination in Hong Kong urban parks and open spaces. Using a hybrid field measurement and computer simulation approach, the study has found that while the air quality in urban parks is better than that of the roadside, it is not significantly different from that of the ambient conditions. Noise levels in the urban parks are not significantly lower than those at the typical home environment in Hong Kong. Heavy metals in urban park dust are slightly lower than that found at the roadside and are broadly comparable to those found at typical homes and nursery schools. These findings lend support to the postulation that the capability of urban parks and open spaces in dense cities to improve the urban environment is rather limited and call for a re-examination of the role of urban parks in enhancing urban livability. The findings also have implications on how urban parks in dense cities should be designed and managed. PMID- 16311823 TI - Higher plants as bioindicators of sulphur dioxide emissions in urban environments. AB - The evaluation of certain vascular plants that grow in the city of Madrid as biomonitors of SO(2) air pollution in urban environments has been carried out. Total concentration of sulphur in leaves of the chosen higher plants as well as other parameters in close relation to this contaminant (visible injury symptoms, chlorophyll a- and b-content and peroxidase activity) have been determined in order to study the spatial distribution and temporal changes in SO(2) deposition. Results obtained show that coniferous species such as Pinus pinea, were more sensitive to SO(2) atmospheric concentration than leafy species as Quercux ilex subspecies ballota and, in the same way, bush species, such as Pyracantha coccinea and Nerium oleander, were more sensitive than wooded species, such as Cedrus deodara and Pinus pinea, respectively. There is a higher accumulation of sulphur in vegetable species located near highways and dense traffic incidence roads and near areas with high density of population. The minimum values for accumulation of SO(2) were registered in winter and spring seasons (from January to April) due to the vegetative stop; while maximum values are obtained during the summer season (from June to September), due to the stoma opening. The highest increments in sulphur concentration, calculated as the difference between two consecutive months, are obtained in May and June for all considered species except for Cedrus deodara and Pyracantha coccinea, both species have few seasonal changes during the whole year. Some species are more sensitive to natural washing than others, showing a decrease in sulphur concentration after rainfall periods. PMID- 16311824 TI - Air quality assessment and control of emission rates. AB - Mathematical methods based on the adjoint model approach are given for the air pollution estimation and control in an urban region. A simple advection-diffusion reaction model and its adjoint are used to illustrate the application of the methods. Dual pollution concentration estimates in ecologically important zones are derived and used to develop two non-optimal strategies and one optimal strategy for controlling the emission rates of enterprises. A linear convex combination of these strategies represents a new sufficient strategy. A method for detecting the enterprises, which violate the emission rates prescribed by a control, is given. A method for determining an optimal position for a new enterprise in the region is also described. PMID- 16311825 TI - Assessment of a natural wetland for use in wastewater remediation. AB - An environmental study was conducted to assess various aspects of the water and sediment quality of a natural wetland to determine its feasibility for advanced wastewater treatment in Louisiana. Nitrate (NO(3)), nitrite (NO(2)), ammonia (NH(4)), total Kjeldahl nitrogen (TKN), total phosphorus, chloride, total organic carbon, pH, trace metals, fecal coliform, dissolved oxygen (DO), and biochemical oxygen demand (BOD) were monitored. Productivity of a dominant shrub, Iva frutescens, in the wetland was also assessed. Research results indicated that gradients of chloride and salinity concentrations showed a broad mixing of the discharged fresh water into the more saline natural brackish waters. This provided an ideal pattern for nutrient assimilation by the receiving marsh. NH(4) was reduced in the range of 50-100% when all combinations of sources and outflows were considered. For total phosphorus and TKN, reduction ranged from 0-95.1% and 11.2-89.7%, respectively. Some nutrient concentrations in the effluent outlet, NO(3) in particular, were lower than background concentrations found in the reference wetland. Sediment and water showed no significant deficiency or toxicity problems for the major nutrients and metals analyzed. The secondary effluent discharges had little demonstrable negative impact on the wetland during the study period from 1995 to 1996. PMID- 16311826 TI - Sensitive parameters in predicting exposure contaminants concentration in a risk assessment process. AB - A sensitivity analysis (SA) was conducted on the analytical models considered in the risk-based corrective-action (RBCA) methodology of risk analysis, as developed by the American Society for Testing of Materials (ASTM), to predict a contaminant's concentration in the affected medium at the point of human exposure. These models are of interest because evaluations regarding the best approach to contaminated site remediation are shifting toward increased use of risk-based decision, and the ASTM RBCA methodology represents the most effective and internationally widely used standardized guide for risk assessment process. This paper identifies key physical and chemical parameters that need additional precision and accuracy consideration in order to reduce uncertainty in models prediction, thereby saving time, money and engineering effort in the data collection process. SA was performed applying a variance-based method to organic contaminants migration models with reference to soil-to-groundwater leaching ingestion exposure scenario. Results indicate that model output strongly depends on the organic-carbon partition coefficient, organic-carbon content, net infiltration, Darcy velocity, source-receptor distance, and first-order decay constant. PMID- 16311827 TI - Sampling the soil in long-term forest plots: the implications of spatial variation. AB - Long-term monitoring of forest soils as part of a pan-European network to detect environmental change depends on an accurate determination of the mean of the soil properties at each monitoring event. Forest soil is known to be very variable spatially, however. A study was undertaken to explore and quantify this variability at three forest monitoring plots in Britain. Detailed soil sampling was carried out, and the data from the chemical analyses were analysed by classical statistics and geostatistics. An analysis of variance showed that there were no consistent effects from the sample sites in relation to the position of the trees. The variogram analysis showed that there was spatial dependence at each site for several variables and some varied in an apparently periodic way. An optimal sampling analysis based on the multivariate variogram for each site suggested that a bulked sample from 36 cores would reduce error to an acceptable level. Future sampling should be designed so that it neither targets nor avoids trees and disturbed ground. This can be achieved best by using a stratified random sampling design. PMID- 16311828 TI - Spatial extent of degraded sediment quality in Puget Sound (Washington State, U.S.A.) based upon measures of the sediment quality triad. AB - A survey was designed and conducted to determine the severity, spatial patterns, and spatial extent of degraded sediment quality in Puget Sound (Washington State, USA). A weight of evidence compiled from results of chemical analyses, toxicity tests, and benthic infaunal analyses was used to classify the quality of sediments. Sediment samples were collected from 300 locations within a 2363 km(2) area extending from the US/Canada border to the inlets of southern Puget Sound and Hood Canal. Degraded conditions, as indicated with a combination of high chemical concentrations, significant toxicity, and adversely altered benthos, occurred in samples that represented about 1% of the total area. These conditions invariably occurred in samples collected within urbanized bays and industrial waterways, especially near the urban centers of Everett, Seattle, Tacoma, and Bremerton. Sediments with high quality (as indicated by no toxicity, no contamination, and the presence of a relatively abundant and diverse infauna) occurred in samples that represented a majority (68%) of the total study area. Sediments in which results of the three kinds of analyses were not in agreement were classified as intermediate in quality and represented about 31% of the total area. Relative to many other estuaries and marine bays of the USA, Puget Sound sediments ranked among those with minimal evidence of toxicant-induced degradation. PMID- 16311829 TI - Application of artificial neural network models to analyse the relationships between Gammarus pulex L. (Crustacea, Amphipoda) and river characteristics. AB - This study aimed at analysing the relationship between river characteristics and abundance of Gammarus pulex. To this end, four methods which can identify the relative contribution and/or the contribution profile of the input variables in neural networks describing the habitat preferences of this species were compared: (i) the "PaD" ("Partial Derivatives") method consists of a calculation of the partial derivatives of the output in relation to the input variables; (ii) the "Weights" method is a computation using the connection weights of the backpropagation Artificial Neural Networks; (iii) the "Perturb" method analyses the effect of a perturbation of the input variables on the output variable; (iv) the "Profile" method is a successive variation of one input variable while the others are kept constant at a fixed set of values. The dataset consisted of 179 samples, collected over a three-year period in the Zwalm river basin in Flanders, Belgium. Twenty-four environmental variables as well as the log-transformed abundance of Gammarus pulex were used in this study. The different contribution methods gave similar results concerning the order of importance of the input variables. Moreover, the stability of the methods was confirmed by gradually removing variables. Only in a limited number of cases a shift in the relative importance of the remaining input variables could be observed. Nevertheless, differences in sensitivity and stability of the methods were detected, probably as a result of the different calculation procedures. In this respect, the "PaD" method made a more severe discrimination between minor and major contributing environmental variables in comparison to the "Weights", "Profile" and "Perturb" methods. From an ecological point of view, the input variables "Ammonium" and to a smaller extent "COD", were selected by these methods as dominant river characteristics for the prediction of the abundance of Gammarus pulex in this study area. PMID- 16311830 TI - Precision and accuracy of pest and pathogen damage assessment in young eucalypt plantations. AB - Fungal pathogens, browsing mammals, birds, insects, nutrient deficiencies, drought, frost and waterlogging are all damaging agents to plantation species. The subsequent loss in leaf tissue or reduced photosynthetic potential can reduce growth and potentially lead to tree death. The Crown Damage Index (CDI) was developed in Australia to quantify damage in young eucalypt plantations. The accuracy and precision of assessing damage at a tree level were determined to ensure the reliability, objectivity and repeatability of the CDI method. Nine assessors, with varying levels of experience, estimated damage on three plots of fifty trees each, to obtain an understanding of the subjectivity of assessing damage caused by insects (e.g. Chrysophtharta spp.) and fungal pathogens (e.g. Mycosphaerella spp.) on Eucalyptus globulus. Damage levels were measured by destructive sampling to enable direct comparisons between estimates and damage levels to be made. The most experienced assessors provided the most repeatable estimates and were generally the most accurate. The incidence of foliar necrosis was the least subjective measure while defoliation was the most subjective and the least accurate of the indices measured. All assessors, regardless of experience, were able to predict the Crown Damage Index (a combined index of all damage classes) to within 12% of measured damage levels. PMID- 16311831 TI - Environmental monitoring of remedial dredging at the New Bedford Harbor, MA, Superfund site. AB - New Bedford Harbor (NBH), MA, is a Superfund site because of high polychlorinated biphenyl (PCB) concentrations in the sediment. From April 1994 to September 1995, a remedial dredging operation (termed the "Hot Spot") removed the most contaminated sediments (PCB concentrations greater than 4000 microg/g) from the upper harbor. During remediation, a monitoring program assessed the potential environmental impacts to NBH and adjacent Buzzards Bay. The monitoring program was developed with input from federal, state, and local authorities. Site specific decision criteria were established to assess net PCB transport, water column toxicity, and PCB bioaccumulation in blue and ribbed mussels (Mytilus edulis and Geukensia demissa, respectively). The remediation was completed without exceeding PCB net transport or acute toxicity effects specified in the decision criteria. In addition, PCB bioaccumulation in mussels during this time period was not significantly greater than pre- or post-operational measurements. The results indicated that approximately 14000 cubic yards of highly PCB contaminated sediment were permanently removed with minimal environmental effects. The lessons learned during this operation, as well as previous pilot studies at the site, will be used to make full-scale remedial efforts in NBH more efficient and environmentally protective. PMID- 16311832 TI - Scatterscore: a reconnaissance method to evaluate changes in water quality. AB - Water quality data collected in periodic monitoring programs are often difficult to evaluate, especially if the number of parameters is large, the sampling schedule varies, and values are of different orders of magnitude. The Scatterscore Water Quality Evaluation was developed to yield a quantitative score, based on all measured variables in periodic water quality reports, indicating positive, negative or random change. This new methodology calculates a reconnaissance score based on the differences between up-gradient (control) versus down-gradient (treatment) water quality data sets. All parameters measured over a period of time at two or more sampling points are compared. The relationship between the ranges of measured values and the ratio of the medians for each parameter produces a data point that falls into one of four sections on a scattergram. The number and average values of positive, negative and random change points is used to calculate a Scatterscore that indicates the magnitude and direction of overall change in water quality. The Scatterscore Water Quality Evaluation, a reconnaissance method to track general changes, has been applied to 20 sites at which coal utilization by-products (CUB) were used to control acid mine drainage (AMD). PMID- 16311833 TI - The water quality of the Certima River basin (Central Portugal). AB - The aim of this study was to evaluate the water quality of the Certima River basin (Central Portugal). For that purpose, surface water samples were collected in March, May and July 2003, at 10 selected sampling sites, and were analysed for physicochemical parameters, namely temperature, conductivity, pH, total suspended solids, dissolved oxygen, biochemical oxygen demand (BOD(5)), Kjeldahl nitrogen and total phosphorus. Results revealed an acceptable water quality during the spring season. Maxima of 64 mg dm(-3) for BOD(5), 39 mg dm(-3) for Kjeldahl nitrogen, and 5.2 mg dm(-3) for total phosphorus, were recorded during summer, indicating a significant degradation of the water quality in a river stretch located downstream of the town of Mealhada. These values, which did not comply with the objectives of minimum quality for surface waters prescribed by the Portuguese legislation, were related to domestic wastewater discharges and runoff waters from a cattle farm. Besides their effects on the middle stretch of the river, these pollution sources were the most likely cause of the high nutrient load in downstream waters, and thus may have a major impact on the trophic status of Pateira de Fermentelos, a sensitive wetland area located in the lower Certima basin. PMID- 16311834 TI - Monitoring the dynamics of coastal vegetation in southwestern Taiwan. AB - This study analyzes the results of the first 5 years of long-term environmental monitoring of the dynamics of coastal vegetation communities in southwestern Taiwan. Seven permanent plots were established in major vegetation communities, including grassland, windbreak forest, and secondary succession forest. Results showed that species richness decreased yearly in grasslands but fluctuated moderately in the forest plots. A Jaccard similarity coefficient was used to evaluate the similarities of species composition between different monitoring years. Species composition changed rapidly in grassland sites, with the similarity coefficient dropping from 82 to 29% in 5 years. The similarity coefficient of vegetation in the composite hardwood forest dropped from 80 to 50%, indicating that at least half the species were the same as those in the beginning and that the composition of forest communities was more stable than that of grassland communities. Dominant species in the forest community changed gradually during the monitoring period. The original planting of Casuarina equisetifolia in windbreak forests decreased year by year in most of the plots, while Cerbera manghas and Ficus microcarpa became the dominant species. The trend of replacement of dominant species indicates that most of the vegetation communities are still in successional stages. PMID- 16311837 TI - Unstable burst fractures of the thoraco-lumbar junction: treatment by posterior bisegmental correction/fixation and staged anterior corpectomy and titanium cage implantation. AB - BACKGROUND: Controversy exists about the best treatment of unstable thoraco lumbar (TL) burst fractures. Kyphosis correction and canal decompression in case of a neurological deficit are recognized treatment objectives, and various conservative and surgical strategies have been proposed. This prospective observational study evaluates the benefits and risks of a posterior bisegmental transpedicular correction/fixation and staged anterior corpectomy and titanium cage implantation in unstable TL junction burst fractures. METHOD: 20 consecutive patients with a single-level traumatic unstable burst fracture at the TL junction were operated on by a bisegmental posterior correction/fixation, followed by anterior corpectomy and titanium cage implantation 7-10 days later. The radiological and clinical course is documented over a period of 24 months. FINDINGS: The mean posttraumatic loss of anterior vertebral body height was 58% (45-70%). The posttraumatic mean regional kyphosis was 16 degrees and could be corrected by the posterior approach to a mean lordosis of 2 degrees. Mean secondary loss of the kyphosis correction was 3 degrees over 24 months. No hardware failure occurred, and construct stability was observed in all 20 patients. One surgical complication occurred during the posterior approach, and three transient surgical complications by the anterior approach. 12 of the 14 patients with an initial neurological deficit recovered an average of 1.5 grades on the ASIA scale. At 24 months postoperatively, the mean regional TL back pain on a VAS (0-10) was 1.6, and the mean pain at the anterior approach site was 1.2. CONCLUSION: Posterior bisegmental transpedicular correction/fixation and staged anterior corpectomy and titanium cage implantation is a safe and reliable surgical treatment option in unstable TL junction burst fractures. The advantages of this technique are a complete kyphosis correction, immediate stability, maintenance of kyphosis correction, and complete spinal canal decompression in case of a neurological deficit. However, these advantages have to be carefully weighed against the double approach morbidity. PMID- 16311838 TI - Acute spinal cord compression in hereditary multiple exostoses. AB - Osteocartilaginous exostoses are benign bone tumors frequently found in the metaphysis of long bones but rarely in the spine. Four patients with acute spinal cord decompensation due to vertebral exostoses spinal cord compression have been previously described in the literature. We report an additional case of rapidly evolving spinal cord compression due to a cervical osteochondroma in a patient with hereditary multiple exostoses (HME), also known as Bessel Hagen disease. Careful analysis of the 5 cases suggested to us that patients with HME should have a systematic spinal imaging screening, in order to prevent rapid neurological decompensation. A minimal risk surgical procedure can be performed at a time of election. PMID- 16311839 TI - Epidermoid tumour of the lateral ventricle. AB - Epidermoids occurring within the lateral ventricles are rare. At one time, they were regarded as anatomical curiosities. The lesions are of developmental aetiology, due to migration of epiblast inclusion at the time of formation of the cerebral vesicle. They are slow growing, and presentation is non-specific in the form of deteriorating mental functions. Generally seen in the fifth decade, but they have also been observed in the paediatric age group. MRI is suggestive of a cystic lesion, and is confirmed to be a typical epidermoid within the lateral ventricle at operation, often having a connection to the midline through the choroidal fissure. The cysts should be excised with no additional morbidity. Histopathology reveals typical stratified squamous epithelium. PMID- 16311840 TI - Early surgery for brainstem cavernomas. AB - BACKGROUND: The purpose was to review our experience with the surgical management of brainstem cavernomas (BSCs) and especially the impact of the surgical timing on the clinical outcome. METHOD: We retrospectively reviewed 22 patients harboring a BSC, who underwent 23 procedures. FINDINGS: Surgery was carried out during the early stage after the last haemorrhage, with a mean delay of 21.6 days (range 4-90 days). Sixteen procedures were performed after a first bleeding event while seven after multiple bleedings. Complete resection was achieved in 19 patients (86.4%). Early after surgery, 12 patients (52.2%) improved neurologically, 5 (21.7%) were stable and 6 (26.1%) worsened. New postoperative deficits were noted after 9 procedures (39.1%). Statistically significant factors for postoperative aggravation were: late surgery (P = 0.046) and multiple bleedings (P = 0.043). No patient operated on within the first 19 days after bleeding did worsen (n = 11), as opposed to 6 out of 12 who did when operated on later. After a mean follow-up of 44.9 months, 20 patients (90.9%) were improved, 1 patient (4.6%) was worse and 1 patient was lost to follow-up (4.6%), after reoperation for rebleeding of a previously completely resected cavernoma. Late morbidity was reduced to 8.6%. The mean Glasgow Outcome Scale (GOS) at the end of the follow-up period was 4.24, compared to a mean preoperative GOS of 3.22 (P<0.001). Complete neurological recovery of motor deficits, sensory disturbances, cranial nerves (CNs), internuclear ophtalmoplegia and cerebellar dysfunction were respectively 41.7%, 38.5%, 52.6%, 60.0% and 58.3%. Among the most affected CNs: CN 3, CN 5 and CN 7 were more prone to completely recover, respectively in 60.0%, 70.0% and 69.2%. CONCLUSIONS: Surgical removal of BSCs is feasible in experienced hands with acceptable morbidity and good outcome. Early surgery and single bleeding were associated with better surgical results. PMID- 16311841 TI - The use of ultrasonic bone curettes in spinal surgery. AB - OBJECT: The author describes a clinical applications, procedure, and efficacy of ultrasonic bone curette (UBC) (bone-removal bar) in spinal surgery. METHODS: From July 2003 to March 2005, we operated on 98 consecutive spinal disorders by using UBC, Sonopet UST-2001; Chiari type-1 malformation (6 cases), cervical disorders (64 cases), thoracic disorders (5 cases), and lumbar disorders (23 cases). The instrument was devoid of any spinning parts, and the risk of grabbing cotton pledgets and damaging normal tissue was thereby avoided. Support from assistants for irrigation and suction was not required. FINDINGS: In this series, there were no iatrogenically induced injuries for example, direct or heat damage to surrounding soft tissues, including nerves, vessels, spinal cord, and dura matter. Considering potential complications of mechanical injuries by ultrasonic technique, intermittent usage and cotton protection during use under an operating microscope are recommended. We found that the ultrasonic device is easy to handle; however, it is time consuming for removing a large amount of bone and ossified lesions. Therefore, we recommend the combined use of this method with standard drilling. CONCLUSIONS: This system appears to be versatile, safe, and efficient in spinal surgery and could be improved by the development of a better handpiece. PMID- 16311842 TI - A systematic review of brain injury epidemiology in Europe. AB - BACKGROUND: The world's literature on traumatic brain injury (TBI) grows annually including new reports on epidemiologic findings from many regions. With the wide variety of reports emphasizing various factors it is useful to compile these findings, hence the objective of this report. Thus, we describe epidemiological factors from European studies largely published in the last 20 years. METHOD: The Medline was searched for TBI related articles from about 1980 to 2003 including terms such as "epidemiology", "head injury", "brain injury" and others. From the research reports identified, we checked references for additional relevant reports and from those reports we abstracted data on TBI incidence, severity, external cause, gender, mortality, prevalence, cost and related factors. RESULTS: Twenty three European reports met inclusion criteria and included findings from national studies from Denmark, Sweden, Finland, Portugal, Germany, and from regions within Norway, Sweden, Italy, Switzerland, Spain, Denmark, Ireland, the U.K. and France. An aggregate hospitalized plus fatal TBI incidence rate of about 235 per 100,000 was derived. Prevalence rate data were not reported from any European country. An average mortality rate of about 15 per 100,000 and case fatality rate of about 11 per 100 were derived. The TBI severity ratio of hospitalized patients was about 22:1.5:1 for mild vs. moderate vs. severe cases, respectively. The percentages of TBI from external causes varied considerably and several reports reported an association of alcohol use with TBI. Outcome or disability findings were mixed and inconsistent. INTERPRETATION: It was difficult to reach a consensus on all epidemiological findings across the 23 published European studies because of critical differences in methods employed across the reports. We highly recommend the development of research guidelines to standardize definitional, case finding, and data reporting parameters to help establish a more precise description and hence utility of the epidemiology of TBI in Europe. PMID- 16311843 TI - Topotecan can compensate for protracted radiation treatment time effects in high grade glioma xenografts. AB - PURPOSE: Several studies reported that prolongation of overall treatment time of fractionated radiotherapy reduces the chance of tumor control. In the present study, we hypothesize that combining topotecan with irradiation could compensate for this detrimental time effect on the radioresponse. Therefore, we investigated the efficiency of different schedules of topotecan (TPT), radiotherapy (RT) or concomitant combination TPT + RT. METHODS AND MATERIALS: Experiments were performed in two human high-grade glioma xenograft models (U87 and GBM Nan1). TPT and RT were delivered at a total dose of 3 mg/kg and 40 Gy, respectively. For the TPT + RT groups, TPT was injected 5 min before radiation. Total radiation doses were delivered in 5, 10, 20, or 30 fractions over 1, 2, 4, or 6 weeks, respectively. The efficiency of TPT, RT, and TPT + RT was evaluated by tumor growth delay (TGD). RESULTS: At this low total dose, and independent of the schedule, no efficacy was found in TPT-treated glioma xenografts. Conversely, radiotherapy-induced antitumor effect decreased with prolongation of treatment time. For TPT + RT combination, antitumor activity was not influenced by schedule, and tumor response was always comparable to those measured for the shortest and the most efficient irradiation schedule (i.e. 1 week). When treatment was delivered over 4 or 6 weeks in U87 glioma xenografts, therapeutic enhancement ratios reached 2.6 and 3.7, respectively. This indicated that the interaction between ionizing radiation and topotecan was synergistic. CONCLUSION: The present study demonstrated that concomitant topotecan can compensate for the detrimental effect of treatment time protraction on radiotherapy efficacy in two malignant glioma xenografts. PMID- 16311844 TI - Navigational maxillofacial surgery using virtual models. AB - Ablative tumor surgery and orbital and midface reconstruction, as much as orthognathic surgery, requires detailed planning using computed tomography (CT) or magnetic resonance imaging (MRI). These techniques also allow simulation of complex surgeries preoperatively. Proper reconstruction depends on reliable information to choose the correct type of grafts and to predict the outcome. This study evaluates the benefit and indications of computer-assisted surgery in the treatment of 107 patients who underwent craniomaxillofacial surgery. Based on a CT or MRI data set, an optical navigation system was used for preoperative planning, intraoperative navigation, and postoperative control. Surgery could be preoperatively planned and intraoperatively navigated. Preoperatively, it required that soft and hard tissues were measured using the mirrored data set of the unaffected side; the size and location of the graft were chosen virtually. Intraoperatively contours of transplanted tissues were navigated to the preoperatively simulated reconstructive result. Computer-assisted treatment was successfully completed in all 107 cases. Preoperatively outlined safety margins could be exactly controlled during tumor resection. Reconstruction was designed and performed precisely as virtually planned. Image-guided treatment improves preoperative planning by visualizing the individual anatomy, outlining the intended reconstructive outcome, and by objectifying the effect of adjuvant therapy. Intraoperative navigation makes tumor and reconstructive surgery more reliable by showing the safety margins, saving vital structures, and leading the reconstruction to preoperatively planned objectives. PMID- 16311845 TI - Needle tract implantation of papillary thyroid carcinoma after fine-needle aspiration biopsy. AB - Although fine-needle aspiration biopsy (FNAB) is a useful tool for diagnosing thyroid carcinoma, there are some risks of complications. In this study, we investigated tumor implantation by FNAB of papillary carcinoma. We compared the characteristics of the main tumors and implanted tumors of patients showing FNAB implantations. Between 1990 and 2002, 4912 patients underwent FNAB and were diagnosed as having papillary carcinoma. We encountered 7 cases of needle tract implantation, which account only for 0.14%. We reviewed these 7 cases as well as 3 other patients who underwent FNAB in other hospitals. The intervals between FNAB and detection of the implanted tumor ranged from 2 to 131 months. For these 10 patients, the main tumors in 6 were diagnosed as poorly differentiated carcinoma, and 7 showed extrathyroid extension. Five showed the development of implanted tumor after comparatively shorter intervals (2-68 months), and we classified these as the short interval group. The remaining 5 were classified as the long interval group, because tumor development occurred after 87-131 months. All 5 cases in the short interval group involved preoperatively detectable lymph node metastasis; those in the long interval group did not. The MIB-1 labeling index of the implanted tumor was high in 4 cases in the short interval group, but it was low in all cases in the long interval group. The implanted tumors could be surgically removed without recurrence at the focal sites. These findings indicate that, although high growth activity in the metastatic lesions may be a risk factor of FNAB, inducing the growth of implanted tumors along the needle tract within a short interval after the procedure, FNAB remains the most useful technique for diagnosing thyroid carcinoma. The incidence of implantation was low, and when it did occur, the tumors could be surgically removed without recurrence. PMID- 16311846 TI - Incisional hernia: early complication of abdominal surgery. AB - It has been suggested that early development of the incisional hernia is caused by perioperative factors, such as surgical technique and wound infection. Late development may implicate other factors, such as connective tissue disorders. Our objective was to establish whether incisional hernia develops early after abdominal surgery (i.e., during the first postoperative month). Patients who underwent a midline laparotomy between 1995 and 2001 and had had a computed tomography (CT) scan of the abdomen during the first postoperative month were identified retrospectively. The distance between the two rectus abdominis muscles was measured on these CT scans, after which several parameters were calculated to predict incisional hernia development. Hernia development was established clinically through chart review or, if the chart review was inconclusive, by an outpatient clinic visit. The average and maximum distances between the left and right rectus abdominis muscles were significantly larger in patients with subsequent incisional hernia development than in those without an incisional hernia (P < 0.0001). Altogether, 92% (23/25) of incisional hernia patients had a maximum distance of more than 25 mm compared to only 18% (5/28) of patients without an incisional hernia (P < 0.0001). Incisional hernia occurrence can thus be predicted by measuring the distance between the rectus abdominis muscles on a postoperative CT scan. Although an incisional hernia develops within weeks of surgery, its clinical manifestation may take years. Our results indicate perioperative factors as the main cause of incisional hernias. Therefore, incisional hernia prevention should focus on perioperative factors. PMID- 16311847 TI - Percutaneous versus Conventional Tracheostomy in Burned Patients with Inhalation Injury. AB - The aim of the study was to compare conventional tracheostomy with percutaneous dilatational tracheostomy in patients with inhalation burn injury. A total of 37 patients with severe burn injuries and associated inhalation injury, underwent percutaneous tracheostomy in our burn unit and were retrospectively compared with 22 patients who underwent conventional surgical tracheostomy. In the first group, 25 of 37 patients and in the second group 17 of 22 patients presented with partial or full-thickness burn injuries (or both) in the neck region. The cost of the procedure, operating time, complications, and incidence of pulmonary infection were recorded. There were no significant perioperative complications in the percutaneous tracheostomy group, and no patient required surgical revision or conversion to surgical tracheostomy. In the conventional tracheostomy group, 2 patients developed tracheal stenosis, 1 had a tracheoesophageal fistula, and 10 had stomal infections. The average procedure time in the first group was 9 minutes, and in the second group it was 22 minutes. The cost of the bedside percutaneous tracheostomy was one-fifth the cost of a conventional tracheostomy. The incidence of pulmonary sepsis was 45% after percutaneous tracheostomy compared to 68% after conventional tracheostomy. With the percutaneous technique, spontaneous closure of the stoma occurred within 1 to 3 days after removal of the tracheostomy tube, whereas with the conventional technique it was within 5 to 7 days. Percutaneous tracheostomy is associated with a lower complication rate and can be safely performed at the bedside. Moreover, it is faster and can be done at a lower cost than conventional open tracheostomy. PMID- 16311848 TI - Escin: inhibiting inflammation and promoting gastrointestinal transit to attenuate formation of postoperative adhesions. AB - Postoperative peritoneal adhesions are common, serious complications of general abdominal and gynecologic surgery that can lead to chronic abdominal pain, intestinal obstruction, and infertility. As yet, there are no ideal drugs that may be prescribed for patients to prevent adhesion formation effectively. In this study the effects of escin, a natural drug, on the various steps of adhesion formation were investigated. The effects of escin on increased vascular permeability induced by acetic acid in a mouse model of acute inflammation, granuloma formation in a subchronic inflammatory rat model, gastrointestinal transit in rats with intestinal paralysis, intestinal motility in postoperative patients, and postoperative adhesion formation in a rat model were observed. It was shown that escin could inhibit acute inflammation and granuloma formation, cause acceleration of gastrointestinal transit, help recover intestinal motility, and attenuate the formation of postoperative adhesions. The findings suggest that escin attenuates the formation of postoperative adhesions by inhibiting inflammation and promoting gastrointestinal transit. Thus it may be concluded that both inhibition of inflammation and increased gastrointestinal motility during the early postoperative period have a positive effect on decreasing the formation of adhesions. PMID- 16311849 TI - Proximal gastrectomy reconstructed by jejunal pouch interposition for upper third gastric cancer: prospective randomized study. AB - Proximal gastrectomy with jejunal pouch interposition (PGJP) has been advocated as an alternative operation for upper third gastric cancer. However, there has been no prospective randomized trial comparing PGJP with total gastrectomy with Roux-en-Y esophagojejunostomy (TGRY). The aim of this study was to compare the short- and medium-term results of PGJP and TGRY in a randomized clinical trial. Fifty-one patients with upper third gastric cancer were randomized to either PGJP (n = 25) or TGRY (n = 26). Outcome measures were postoperative complications, nutritional status assessed by serum nutritional parameters, and postgastrectomy symptoms. There were no significant differences in operating time, hospital stay, and postoperative complications. Blood loss was significantly less in the PGJP group (P = 0.036). Nineteen patients (73%) in the TGRY group had one or more postgastrectomy symptoms, which was significantly more frequent than in the PGJP group (32%; P = 0.012). There were also significant differences between the two groups with regard to food intake, weight recovery, hemoglobin, and serum vitamin B12 levels in favor of PGJP. In conclusion, proximal gastrectomy with jejunal pouch interposition for upper third gastric cancer is safe, and is associated with a greater reduction in postgastrectomy symptoms and better nutritional status compared with conventional total gastrectomy. PMID- 16311850 TI - Prognostic significance of vascular endothelial growth factor D in gastric carcinoma. AB - The angiogenic factor called vascular endothelial growth factor (VEGF)-D is a ligand for VEGF receptor-2 (VEGFR-2/KDR) and receptor-3 (VEGFR-3/Flt-4). It is implicated in the development of lymphatic vessels and promotion of lymphatic metastasis. The purpose of this study was to investigate the prognostic significance of VEGF-D expression in patients with gastric carcinoma. We assessed the expression of VEGF-D in gastric carcinoma by immunohistochemistry on 143 consecutive patients' stored sections and evaluated the lymphatic vessel count (LVC) in tumors using the novel selective lymphatic endothelium marker D2-40. VEGF-D expression was observed in 55 (39%) tumor sections. The expression of VEGF D correlated significantly with tumor size, T of the TNM classification, lymphatic and venous system invasion, LVC, lymph node metastasis, M of TNM, and pTNM stage. Multivariate analysis indicated that VEGF-D expression was an independent prognostic factor for both relapse-free survival (RFS) and overall survival (OS). Our data indicate the involvement of VEGF-D in tumor progression via lymphoangiogenic pathways. Practically, VEGF-D expression can be useful for predicting RFS and OS in patients with gastric carcinoma. PMID- 16311851 TI - Gender differences in quality of life of patients with rectal cancer. A five-year prospective study. AB - To determine how quality of life changes over time and to assess gender-related differences in quality of life of rectal cancer patients we conducted a 5-year study. Little is known about how quality of life (QoL) changes over time in patients after surgery for rectal cancer, and whether gender of the patients is associated with a different perception of QoL. The aim of this study was to assess prospectively, changes in quality of life after surgery for rectal cancer, with a focus on gender related differences. Over a 5-year period, the EORTC-QLQ-C 30 and a tumor-specific module were prospectively administered to patients before surgery, at discharge, 3, 6, 12, and 24 months postoperatively. Comparisons were made between female and male patients. A total of 519 patients participated in the study, 264 men and 255 women. The two groups were comparable in terms of surgical procedures, adjuvant treatment, tumor stage, and histology. Most QoL scores dropped significantly below baseline in the early postoperative period. From the third month onward, global health, emotional and physical functioning, improved. Female gender was associated with significantly worse global health and physical functioning and with higher scores on treatment strain and fatigue. Men reported difficulties with sexual enjoyment; furthermore, over time, sexual problems created high levels of strain in men, worse than baseline levels in the early postoperative period. These problems tended to continue over the course of time. The findings in this study confirm that QoL changes after surgery and differs between men and women. Women appear to be affected by impaired physical functioning and global health. Female gender is associated with significantly higher fatigue levels and increased strain values after surgery. Through impaired sexual enjoyment, men are put more under strain than woman. PMID- 16311852 TI - Results of surgical, laparoscopic, and percutaneous treatment for hydatid disease of the liver: 10 years experience with 355 patients. AB - Selection of the most appropriate treatment to obtain the best results with the lowest rate of recurrence and minimal morbidity and mortality is mandatory for the management of hepatic hydatid disease. The surgical approach is the mainstay of treatment, and there has been a tendency toward laparoscopic surgery and, more recently, percutaneous treatment (PT), which has become increasingly popular with revolutions in techniques. We aimed to evaluate the results of current therapeutic methods in the context of a 10-year single-institution experience. Between 1992 and 2003, 355 patients with 510 hydatid cysts of the liver were treated by open operation, laparoscopic surgery, or PT. The series included 128 females and 227 males ranging in age from 10 years to 73 years. Preferred treatment modalities, perioperative complications, interventions, recurrences, and length of hospital stay were retrospectively analyzed. There were two postoperative deaths (1.08%) in the open surgery group. Biliary leakage was observed in 28 patients treated with open surgery, in 10 patients after PT, and in 2 after laparoscopic treatment. Recurrence rates were 16.2%, 3.3%, and 3.5% after open surgery, laparoscopic surgery, and percutaneous treatment, respectively. Characteristics of the cyst, presence of cystobiliary communications, and the availability of a multidisciplinary team are the factors that we believe directly affect the results. Radical surgery can be done safely for suitable cases; conventional procedures are associated with greater morbidity. Laparoscopic surgery seems effective and safe, with low morbidity and recurrence rates for type I-III cysts in accessible localizations. Our experience with PAIR (puncture, aspiration, injection, and reaspiration) and catheterization provides evidence that the procedure is an effective and safe option. PMID- 16311853 TI - Outcome in right living related liver transplantation with branch-patch arterial reconstruction. AB - cRight lobe living liver transplantation is being performed worldwide with increased frequency. Difficult arterial reconstructions are often encountered because of small diameter or discrepancy between arterial stumps. The risk of arterial thrombosis is reported as high as 26%: microsurgical techniques have reduced this rate below 2%, increasing warm ischemia time. We have developed a new branch patch technique in living related liver transplantation using the donor cystic artery to create an enlarged patch anastomosis that enables increase in the vessel's diameter and therefore greater inflow to the liver. We have followed 8 patients treated with this technique. After more than 1 year (mean follow-up: 636 days) we did not observe any arterial thrombosis by Doppler ultrasound performed every 3 months. The mean resistance index was 0.68 (0.57 0.83-). Three patients died with functional graft without signs of thrombosis. We believe that the cystic artery branch patch technique is feasible in all cases. It is fast (mean time: 6.2 min), it allows a shorter warm ischemia time, and there is no increased risk of thrombosis. PMID- 16311856 TI - Comparison of resected and non-resected intraductal papillary mucinous neoplasms of the pancreas. AB - By comparing the clinicopathological features and survivals between the resected and non-resected intraductal papillary mucinous neoplasms (IPMNs) of the pancreas, this study tried to clarify the natural history of IPMNs, to provide a strategy for treatment, and to determine the justification of not performing resection for some patients. A total of 57 patients with IPMN, including 39 resected and 18 non-resected IPMNs, were recruited for study. Data on demographics, clinical presentations, diagnostic work-up, treatment modality, clinical course, and outcomes were evaluated and compared between the resected and non-resected IPMNs. The most common clinical presentation was abdominal pain (57% in total IPMNs, 67% in resected, 33% in non-resected), followed by body weight loss (32% in total IPMNs, 33% in resected, 28% in non-resected). The sensitivity in the diagnosis of IPMN was highest by magnetic resonance cholangiopancreatography (MRCP) (88%), followed by endoscopic retrograde cholangiopancreatography (ERCP) (68%), and computed tomography scan (CT scan) (42%) and sonography (10%). The median survival was 21.5 months for patients with resected IPMNs, ranging from 2 to 124 months, and 14 months in non-resected IPMN patients, ranging from 5.5 to 70 months. There is no significant survival difference between the resected and non-resected groups, with a 5-year survival of 69.8% in resected IPMNs and 59.8% in non-resected IPMNs, P = 0.347. The survival outcome of the unresectable non-resected IPMNs was much inferior to the resected IPMNs, P = 0.002 and resectable non-resected IPMNs, P = 0.001. Thus, the prime prognostic factor in predicting the survival outcome of IPMNs is resectability, instead of resection itself. Long-term survival could also be expected in resectable IPMNs without resection. No resection for the IPMN may be justified for patients with high surgical risks, especially for those who are asymptomatic and very aged. PMID- 16311857 TI - Incidental parathyroidectomy during thyroid surgery: an underappreciated complication of thyroidectomy. AB - The aim of this study was to investigate the incidence, risk factors, and clinical relevance of incidental parathyroidectomy during thyroid surgery. Prospective analysis of data in patients following thyroidectomy, specifically regarding the presence of parathyroid parenchyma in the thyroidectomy specimens, the underlying thyroid pathology, and the presence of postoperative hypocalcemia (biochemical/clinical). The clinical records of 158 patients who underwent thyroid surgery during a 2-year period were reviewed. Pathology reports were carefully reviewed for the nature of the underlying thyroid disease, the presence, number, and size of incidentally resected parathyroid gland(s), their location, and possible parathyroid pathology. Serum calcium levels were measured preoperatively, on the day of surgery, and on postoperative days 1, 2, and 7 or even later as needed. Two groups of patients were studied: a group with incidental parathyroidectomy following thyroidectomy (group A) and a group without incidental parathyroidectomy after thyroidectomy (group B). Total/near total thyroidectomy was the procedure of choice and was performed in 154 patients; total lobectomy and contralateral subtotal lobectomy was performed in the other 4 patients. Elective central neck lymph node dissection was performed in four patients with neck lymphadenopathy. Inadvertently removed parathyroid tissue was found in 28 cases (17.7 %); in 6 of these patients (21%) the parathyroid tissue was intrathyroidal. The percentage of women in group A was significantly higher than in group B (93% vs. 58.5%, P = 0.0002). There was no statistically significant difference between the two groups (A and B) regarding the preoperative (presumed) diagnosis, the histologic diagnosis of thyroid disease (benign versus malignant), the type/extent of surgery, or the presence of thyroiditis. Biochemical and clinical hypocalcemia was observed in 6 (21%) and 2 (7%) patients in group A, respectively, and in 30 (23%) and 8 (6%) patients of group B, respectively. There was no statistically significant difference regarding the occurrence of postoperative hypocalcemia (clinical/biochemical) between the two groups (P = 0.33). Incidental parathyroidectomy is not uncommon following thyroidectomy and in a significant percentage of cases it may be due to the intrathyroidal location of the parathyroid glands. Incidental parathyroidectomy was not found to be associated with postoperative hypocalcemia (biochemical/clinical). Incidental parathyroidectomy may be considered as a potentially preventable but clinically minor complication of thyroid surgery. PMID- 16311859 TI - I read with interest and compassion the article by Dr. M. Schein: "The rural surgeon: a new forum in the world journal of surgery". PMID- 16311860 TI - Laparoscopy-assisted gastrectomy for early gastric cancer in young and elderly patients. AB - Open gastric surgery in elderly patients is associated with higher morbidity and mortality rates than those reported among younger individuals. Therefore, minimally invasive surgery may have a larger impact on the elderly compared to the younger age group. The objective of this study was to evaluate the experience of laparoscopy-assisted distal gastrectomy (LADG) in patients with early gastric cancer and compare the results in patients 70 years of age and older to those in patients younger than 70 years of age. From January 1998 to October 2004, a total of 103 patients underwent LADG. Of these patients, 30 who were older than 70 years were compared with 73 who were younger. Preoperative co-morbidity, operative results, postoperative outcomes, and survival were analyzed. Furthermore, as a standard control of this study, we reviewed 54 distal gastrectomy cases with open surgery (open distal gastrectomy; ODG) in the same term with the same background factors, categorized into elder (n = 16) and younger (n = 38). The mean age of the elderly patients was 75 years in the LADG group. A significantly higher proportion of elderly patients had concurrent diseases in both groups. Blood loss was significantly less in the elderly than in younger patients undergoing LADG, and it was less in the LADG group than in the ODG group. The overall 5-year survival rates in the LADG group were not significantly different between elderly and younger patients. Laparoscopy assisted distal gastrectomy is a safe and effective treatment for early gastric cancer in the elderly. Therefore, chronological age alone should not be considered a contraindication in selecting patients for LADG. PMID- 16311865 TI - We read with interest the article by Brauckhoff et al., in the December 2004 issue of the World Journal of Surgery. PMID- 16311866 TI - Pancreatic anastomoses after pancreaticoduodenectomy: do we need further studies? AB - Pancreatic anastomotic leak is the single most important factor responsible for the considerable morbidity and mortality associated with pancreaticoduodenectomy. Management of the pancreatic remnant is controversially discussed, reflecting the complexity of anastomosing a pancreas of different textures to the digestive tract. A number of studies evaluating diverse options have often provided conflicting conclusions. This information is confusing particularly to those surgeons outside of large-volume centers with broad experience and to general surgeons who perform pancreatic surgery. A PubMed search with the key words pancreaticoduodenectomy, pancreatic anastomosis, pancreaticojejunostomy, pancreaticogastrostomy, and pancreatic fistula was performed. Major series of pancreatic anastomosis published between 1990 and 2002 were studied from diverse centers worldwide. Their results with regard to pancreatic fistula, morbidity, and mortality were documented. Nine series of pancreaticojejunostomy and seven series of pancreaticogastrostomy were evaluated. Eight comparative studies evaluating the two techniques were also analyzed. A single randomized controlled trial was identified among these comparative studies. Equally good results were observed with the two techniques. Other uncommon methods of management of the pancreatic remnant (duct occlusion and ligation) were also evaluated. Pancreaticojejunostomy followed by pancreaticogastrostomy are the most favored techniques. A duct-to-mucosa anastomosis is preferred over other methods. Fistula rates of less than 5%-10% should be the standard irrespective of the technique used. Unlike in the past, mortality can be reduced even in the event of an anastomotic dehiscence, and this aspect is primarily dependent on a meticulous anastomosis based on sound surgical principles rather than the method per se. Based on the information accumulated, adherence to these specific principles could ensure a safe and reliable pancreatic anastomosis with mimimal morbidity and mortality after pancreaticoduodenectomy, even in the hands of general surgeons operating outside high-volume centers. PMID- 16311867 TI - One-stage transthoracic operation for the treatment of right lung and liver hydatid cysts. AB - In this study we reviewed our experience of hydatid disease of the lung and the liver and discussed the safety and the follow-up results of the one-stage operation. Between 1990 and 2004, 142 patients with pulmonary hydatid disease underwent operation in our clinic. Of these, 27 (19%) patients had cysts located on the dome of the liver, treated with phrenotomy through a right thoracotomy. Hydatid cysts located in the lungs were managed by means of cystotomy. For liver cysts, cystotomy and the inversion of the cavity with sutures was the surgical method of choice, and a drain was left in place. The pulmonary cysts of 12 (8.4%) patients were bilateral and 5 (3.5%) patients had prior surgical treatment of hepatic (n = 1) or pulmonary (n = 4) hydatid cysts. The liver cysts were approached transdiaphragmatically after the lung cysts were excised in 27 (19%) patients. In patients with pulmonary cysts, cystotomy, with or without capitonnage was performed on 123 (86.6%) patients, and wedge resection was performed on 11 (7.7%), segmentectomy was performed on 6 patients (4.2%), and lobectomy was performed on 2 (1.4%) patients. There was no mortality, and only a small number of complications were encountered: empyema in 3, excessive biliary drainage in 2, and bronchopleural fistula in only 1. We suggest that the extraction of pulmonary and hepatic cysts simultaneously through the transthoracic route is a useful and safe surgical technique. This technique also prevents the need for a second operation. PMID- 16311869 TI - Liver regeneration and splenic enlargement in donors after living-donor liver transplantation. AB - Liver regeneration after donor hepactectomy offers a unique insight into the process of liver regeneration in normal livers. As the liver restores itself, concurrent splenic enlargement occurs. There are many theories about why this phenomenon takes place: some investigators have proposed a relative portal hypertension that leads to splenic congestion or, perhaps, the presence of a common growth factor that induces both the liver and spleen to enlarge. Between the months of June 2001 and May 2004, 112 live donor liver transplants (LDLTs) were performed in Chang Gung Memorial Hospital, Kaohsiung, Taiwan. The total number of donor hepatectomies performed during this period was 113, however, because one of the cases required dual donors. Of our 113 donors, we eventually analyzed the data of 109; 4 patients were lost to follow-up 6 months later and were excluded from our study. The average age of our donor population was 32.32 +/- 8.48 years. The mean liver volume at donation was noted to be 1207.72 +/- 219.95 cm3, and 6 months later, it was 1027.18 +/- 202.41 cm3. Expressed as a percentage of the original volume, the mean liver volume 6 months after hepatectomy was 90.70% +/- 12.47% in this series. For right graft donors, mean liver volume after 6 months was 89.68% +/- 12.37% of the original liver volume, whereas that for left graft donors was 91.99% +/- 12.6%. Only 26 of the 109 (23.85%) donors were able to achieve full regeneration 6 months post-donation. Notably, liver function profiles of all donors were normal when measured 6 months after operation. The average splenic volume at donation as measured by computed tomography (CT) volumetry was 159 +/- 58 cm3, and the splenic volume 6 months post-donation was 213 +/- 85 cm3. There was a mean increment in splenic volume of 35% +/- 28% 6 months after donation. The blood profiles of the donors were monitored; particular attention was given to platelet levels and liver function tests, and these were found to be within normal limits 6 months after operation. Of note, splenic enlargement was significantly greater among right-sided donors than their left-sided counterparts. Greater splenic enlargement was also observed in those donors who achieved full liver regeneration at their evaluation 6 months postoperatively than in those who did not. Although original liver volume was not re-established in most patients 6 months after liver donation, there seemed to have been no untoward effects to the donor. The factors that affect liver regeneration are complex and myriad. Although there is splenic enlargement at 6 months post-donation in donors of LDLT, there are no untoward effects of this enlargement. PMID- 16311870 TI - Mirizzi syndrome: ten years experience from a teaching hospital in Riyadh. AB - OBJECTIVE: Mirizzi syndrome (MS) is an uncommon presentation of cholelithiasis. This study aims to find the incidence and analyze the outcome of management of this condition at Riyadh Medical Complex (RMC) with particular reference to diagnostic methods and outcome of surgical treatment. METHODS: Retrospective study on 17 consecutive patients of MS diagnosed and managed at RMC over ten year period. The records were reviewed for demography, clinical presentation, diagnostic methods, operative procedures, postoperative complication and follow up. RESULTS: The incidence of MS syndrome was 0.7% of 2415 cholecystectomies. There was preponderance of Type I variety (58.8%). Ultrasonography was able to diagnose 82% cases. ERCP suggested the diagnosis in all cases and helped further in classifying and management of these patients. All Type I cases were managed with partial cholecystectomy, two underwent laparoscopic surgery. Three Type II patients were managed by partial cholecystectomy alone. Three patients with Type III variety had choledochoplasty whereas one remaining patient with Type IV variety underwent hepatico-jejunostomy. All patients had complete recovery with 17.6% procedure-related morbidity and no hospital mortality. All patients are doing well over a mean follow up 6.5 years. CONCLUSION: Preoperative diagnosis of Mirizzi syndrome by ultrasound and ERCP is essential to prevent serious complications during surgery. Partial cholecystectomy is an adequate procedure for Types I & II MS. Choledochoplasty provides an effective surgical repair in Type III cases. Although laparoscopic cholecystectomy in MS may be hazardous, it may still be tried in preoperatively diagnosed type I cases, provided the surgeon is experienced and keeps a low threshold for conversion open surgery. PMID- 16311873 TI - The relationship among negative affect, distress tolerance, and length of gambling abstinence attempt. AB - The present study tested the theory that negative affect and one's ability to tolerate distress is associated with failure to quit gambling during an abstinence attempt. Specifically, 16 current pathological gamblers who had at least one sustained period of gambling abstinence lasting a minimum of 3 months (i.e., delayed relapsers) and 16 current pathological gamblers who had never remained abstinent for a period longer than 2 weeks (i.e., immediate relapsers), were assessed for baseline levels of negative affect and stress reactivity, as well as faced with a psychological (mental arithmetic) and physical (breath holding) stressor. Compared to the delayed relapsers, the immediate relapsers displayed higher levels of negative affect and stress reactivity. Immediate relapsers also were less likely to persist on the psychological stressor, suggesting that one's ability to tolerate the initial discomfort of an abstinence attempt may play an important role in gambling treatment outcome. PMID- 16311874 TI - The "locals" market: an emerging gaming segment. AB - One of the newest customer segments to be identified by casino marketers is the "locals" market, or local resident gambler market. This study addresses the relative lack of empirical data on the "locals" market segment. The purposes of the study are to identify important reasons for visiting a particular casino; to identify which casinos respondents visit most often and why; to gain an understanding of such behavioral variables as gambling budget, time spent gambling, and favorite game; and to determine perceptions of different casinos in the Las Vegas area. The results may not generalize across all casino markets but they do provide insight into gamblers' activities and behavior patterns. A telephone survey administered to residents of the Las Vegas metropolitan area yielded a sample of 637 participants who gamble for a least 15 minutes once every 2 months in a legalized gambling establishment. A customer satisfaction index based on casino performance on importance variables is presented. PMID- 16311875 TI - Development of the Drake Beliefs about Chance inventory. AB - The present research describes the development and validation of a cognitive assessment instrument, the Drake Beliefs about Chance (DBC) inventory, designed to determine and quantify erroneous beliefs about games of chance. Principal components analyses showed that the DBC assesses two primary dimensions, Illusion of Control and Superstition. Correlation analyses showed that scores on these two dimensions are related to higher frequency of gambling behaviors in both adults from the general population and clients from gambling treatment centers. Of the two sets of erroneous beliefs, Illusion of Control was a better predictor of gambling than Superstition. This investigation provides additional evidence that participants in games of chance such as casino gamblers possess certain classes of erroneous beliefs regarding the games they play. PMID- 16311876 TI - Reliability and validity of the pathological gambling adaptation of the Yale Brown Obsessive-Compulsive Scale (PG-YBOCS). AB - The Yale Brown Obsessive Compulsive Scale adapted for Pathological Gambling (PG YBOCS) was developed to measure the severity and change in severity of pathological gambling symptoms. The PG-YBOCS is a 10-item clinician-administered questionnaire that measures the severity of PG over a recent time interval (usually within the past one/two week(s)). In order to assess and validate the scale, it was administered to 337 subjects: 188 pathological gamblers and 149 healthy controls. Internal consistency and correlations between individual items and total score were assessed for various permutations of the sample. Other scales were administered to assess convergent, discriminant and content validity. Sensitivity to change was evaluated in treatment studies with fluovoxamine, lithium, and valproate. Each item was frequently endorsed across a range of severity. Good inter-rater reliability and internal consistency were obtained. The PG-YBOCS showed high validity and reliability for total score, item-total correlations, and for each subscale (Thoughts/Urges and Behavior). PG-YBOCS scores correlated with global severity and South Oaks Gambling Screen (SOGS) scores. The scale was also sensitive to change in pathological gambling severity. PG-YBOCS thus appears to be a reliable and valid measure of pathological gambling severity, and can be regarded as an important tool for clinicians and researchers treating pathological gamblers. PMID- 16311877 TI - Gambling, exchange systems, and moralities. AB - History and ethnography show us that, across societies of the past and present, gambling varies considerably with respect to its organization, social meanings, and how it is regarded in moral terms. This paper presents a basic scheme for analyzing the relationship between gambling and society. A theoretical starting point is that reciprocity is fundamental to social and economic systems. An anthropological theory of exchange systems makes a broad distinction between a structural dimension (generalized versus balanced reciprocity) and a normative dimension (from voluntary to involuntary). A model of four basic forms of reciprocity, each having a characteristic exchange mode and morality, can thus be constructed. Gambling is here understood as an exchange system embedded in the reciprocal orders of society and having a necessary relationship to these; it can take on the characteristics of such an order or it can be regarded as conflicting with it. Much of the variation in the form and morality of gambling therefore emerges as systematic and explainable by a theory of forms of reciprocal exchange. PMID- 16311878 TI - The role of perceived control and overconfidence in pathological gambling. AB - Two studies sought to determine whether perceived control has different effects on confidence assessment and betting decisions among pathological and problem gamblers than among non-problem gamblers. In Study 1, 200 college students who were frequent gamblers (80 female and 120 male, median age 20) completed the South Oaks Gambling Screen (SOGS) and then engaged in a task in which they answered questions, assessed confidence in each answer, and considered bets on their answers that were fair if they were well-calibrated, but unfavorable if they were overconfident. Probable pathological and problem gamblers earned significantly fewer points than non-problem gamblers. This was due to greater overconfidence among pathological and problem gamblers, which led to systematically less favorable bets. In Study 2, using 384 participants (105 female and 279 male, median age 20), control was independently manipulated and bets were constructed to make point value independent of overconfidence. Problem and pathological gamblers showed both greater overconfidence and greater bet acceptance. They were less affected by control in their betting decisions than non-problem gamblers, but more affected in the slope of their betting function. It is concluded that pathological and problem gamblers process information about confidence and control differently from non-problem gamblers. PMID- 16311879 TI - Structural changes to electronic gaming machines as effective harm minimization strategies for non-problem and problem gamblers. AB - This study aimed to evaluate the effectiveness of three proposed modifications to the structural characteristics of electronic gaming machines as harm minimisation strategies for non-problem and probable problem gamblers. Structural changes included reducing the maximum bet size, reducing reel spin and removing large note acceptors. Behavioural patterns of play were observed in 779 participants attending clubs and hotels. Observations were conducted in the gaming venue during regular gaming sessions. Eight experimental machines were designed to represent every combination of the modifications. 210 participants played at least one modified and one unmodified machine. Following play, the South Oaks Gambling Screen (SOGS) was administered. More problem than non-problem gamblers used high denomination bill acceptors and bet over one-dollar per wager. Machines modified to accept the one-dollar maximum bet were played for less time and were associated with smaller losses, fewer individual wagers and lower levels of alcohol consumption and smoking. It was concluded that the reduction of maximum bet levels was the only modification likely to be effective as a harm minimization strategy for problem gamblers. PMID- 16311880 TI - Gambling screens: does shortening the time frame affect their psychometric properties? AB - Gambling screening tools such as the South Oaks Gambling Screen (SOGS) and a DSM IV Screen for Gambling Problems (NODS) developed by the National Opinion Research Council have psychometrically validated lifetime and past-year versions. As research questions often dictate shorter time intervals, researchers have adapted the time frames of these instruments to their specific purposes without examining whether changing the time frame affects the psychometric properties. In this study, 3-month versions of SOGS and NODS were administered to a sample of 80 pathological gamblers (59 men, 21 women, mean age 44) seeking treatment at a state-funded facility. The 3-months versions had good internal consistency, good convergent validity with each other, with gambling behaviors assessed via the timeline followback method, and with measures of impulsivity. The 3-month versions also showed good discriminant validity with demographic variables and a measure of verbal IQ. Together the data indicate that shortening the time frame to 3 months does not seem to have adverse effects on the psychometric properties of SOGS and NODS. Thus these adapted versions could profitably be used for shorter time intervals, including as pre/post-treatment and follow-up measures in treatment outcome studies. PMID- 16311881 TI - Gambling and problem gambling among recently sentenced male prisoners in four New Zealand prisons. AB - Recently sentenced inmates in four New Zealand male prisons (N = 357) were interviewed to assess their gambling involvement, problem gambling and criminal offending. Frequent participation in and high expenditure on continuous forms of gambling prior to imprisonment were reported. Nineteen percent said they had been in prison for a gambling-related offence and most of this offending was property related and non-violent. On the basis of their SOGS-R scores, 21% were lifetime probable pathological gamblers and 16% were probable pathological gamblers during the six months prior to imprisonment. Of the "current" problem gamblers, 51% reported gambling-related offending and 35% had been imprisoned for a crime of this type. Gambling-related offending increased with problem gambling severity. However, only five percent of problem gamblers said their early offending was gambling-related. The large majority reported other types of offending at this time. Few men had sought or received help for gambling problems prior to imprisonment or during their present incarceration. This highlights the potential for assessment and treatment programs in prison to reduce recidivism and adverse effects of problem gambling and gambling-related offending. PMID- 16311882 TI - Gambling and problem gambling among recently sentenced women in new zealand prisons. AB - Ninety-four recently sentenced women prisoners were interviewed to assess aspects of their gambling involvement, problem gambling and relationships between gambling and criminal offending. A third of the women, on the basis of their SOGS R scores, were assessed as lifetime probable pathological gamblers and just under a quarter were assessed as probable pathological gamblers during the 6 months prior to imprisonment. For women prisoners, a preference for non-casino gaming machines and housie were predictive of problem gambling. Relative to non-problem gamblers, problem gamblers experienced higher rates of childhood conduct disorder and current non-psychotic mental disorder. Just over a quarter of prisoners and a half of the problem gamblers had committed a crime to obtain money to gamble. Few women said their early offending or convictions related to gambling. It was concluded that most women were "criminals first and problem gamblers second" rather than people whose offending careers commenced as a consequence of problem gambling. However, the extent of problem gambling-related offending among the women prisoners highlights the potential for comprehensive assessment and treatment programs in prison to reduce recidivism and other adverse impacts of problem gambling and gambling-related offending. PMID- 16311883 TI - Serum hyaluronidase aberrations in metabolic and morphogenetic disorders. AB - Hyaluronidases are endo-glycosidases that degrade both hyaluronan (hyaluronic acid) (HA) and chondroitin sulfates. Deficiency of hyaluronidase activity has been predicted to result in a phenotype similar to that observed in mucopolysaccharidosis (MPS). In the present study, we surveyed a variety of patients with phenotypes similar to those observed in MPS, but without significant mucopolysacchariduria to determine if some are based on aberrations in serum hyaluronidase (Hyal-1) activity. The study included patients with well characterized dysmorphic disorders occurring on genetic basis, as well as those of unkown etiology. The purpose of the study was to establish how wide spread were abnormalities in levels of circulating Hyal-1 activity. A simple and sensitive semi-quantitative zymographic procedure was used for the determination of activity. Levels of both beta-N-acetylglucosaminidase and beta-glucuronidase whose activities contribute to the total breakdown of hyaluronan (HA) were also measured, as well as the concentration of circulating HA. Among 48 patients with bone or connective tissue abnormalities, low levels of Hyal-1 activity were found in six patients compared to levels in 100 healthy donors (2.0-3.2 units/microL vs 6(+/- 1 SE) units/microL). These six patients exhibited a wide spectrum of clinical abnormalities, in particular shortened extremities: they included three patients with unknown causes of clinical symptoms, one patient with Sanfilippo disease, one of the seven patients with achondroplasia, and one with hypophosphotemic rickets. Normal levels of serum Hyal-1 activities were found in patients with Morquio disease, GM1 gangliosidosis, I cell-disease, 6 of the 7 patients with achondroplasia, Marfan's-syndrome and Ehlers-Danlos syndrome. No patient totally lacked serum Hyal-1 activity. Serum HA concentration was elevated in patients with Sanfilippo A and I-cell disease. Determination of serum and leukocyte Hyal-1 and serum HA may be useful to evaluate patients with metabolic and morphogenetic disorders. PMID- 16311884 TI - Platelet derived growth factor recruits lactosylceramide to induce cell proliferation in UDP Gal:GlcCer: beta1 --> 4Galactosyltransferase (GalT-V) mutant Chinese hamster ovary cells. AB - Recent molecular cloning studies have suggested the presence of at least two beta4Gal transferase genes (beta4GalT-V and beta4GalT-VI) that may encode lactosylceramide synthase but whether they are functional in vivo and whether they mediate growth factor induced phenotypic change such as cell proliferation is not known. Our previous studies lead to the suggestion that various risk factors in atherosclerosis such as oxidized LDL, shear stress, nicotine, tumor necrosis factor-alpha converge upon LacCer synthase to induce critical phenotypic changes such as cell proliferation and cell adhesion. However, whether platelet derived growth factor also recruits LacCer synthase in mediating cell proliferation is not known. Here we have employed a Chinese hamster ovary mutant cell line Pro(-)5Lec20 to determine whether this enzyme physiologically functions to mediate cell proliferation. We show that PDGF stimulates the activity of UDP galactose:glucosylceramide, beta1,4galactosyltransferase. The activity of LacCer synthase increased about 2.5 fold within 2.5-5 min of incubation with PDGF in both wild type and Pro(-)5Lec20 cells. Concomitantly, there was an increase in the generation of superoxide radicals, p44MAPK phosphorylation and cell proliferation in CHO cells. D-phenyl-2-decanoylamino-3-morpholino-1-propanol (D PDMP), a potent inhibitor of GlcCer synthase/LacCer synthase impaired PDGF mediated induction of LacCer synthase activity, superoxide generation, p44 MAPK activation and cell proliferation in Pro(-)5Lec20 cells. PDGF-induced superoxide generation was also mitigated by the use of diphenylene iodonium; an inhibitor of NADPH oxidase activity that is required for superoxide generation. This inhibition was bypassed by the addition of lactosylceramide. Thus, beta4GalT-V gene produces a bona fide LacCer synthase that can function in vivo to generate LacCer. Moreover, this enzyme alone can mediate PDGF induced activation of a signal transduction cascade involving superoxide generation, p44MAPK activation, phosphorylation of Akt and cell proliferation. PMID- 16311885 TI - Identification of 9-O-acetyl-N-acetylneuraminic acid in normal canine pre-ocular tear film secreted mucins and its depletion in Keratoconjunctivitis sicca. AB - O-Acetylated sialic acids have been reported in many sialoglycoproteins where they mediate a variety of immune and other biological events. We have previously demonstrated that the protective mucus barrier on the surface of the canine eye contains sialoglycoproteins. We have also investigated the occurrence of O acetylated sialic acids in these ocular mucins. Mucus aspirated from the surface of normal dog eyes and those with keratoconjunctivitis sicca (KCS) was fractionated into three pools by density gradient centrifugation. Sialic acids comprised 0.6-0.9% of the dry weight of the mucins isolated. The sialic acid profile in these pools was examined using HPLC. O-Acetylated sialic acids, mainly Neu5,9Ac2, were detected in normal animals and made up 10-30% of the total sialic acids detected. A doubling of the sialic acid content was found in KCS mucins, but the level of 9-O-acetylated sialic acid was reduced below 4% of total. Histological analysis of conjunctival tissue from normal and KCS dogs showed the presence of sialic acids, detected with the alpha(2-6) sialic acid-specific lectin Sambucus nigra, in the goblet cells and corresponding to the staining pattern for MUC5AC, the major ocular-secreted mucin gene product. In KCS animals a disruption of the normal pattern of conjunctival goblet cells was seen with preservation of the pattern of lectin binding observed in normal animals. Thus the data demonstrate the presence of mono-O-Acetylated sialic acids in normal canine ocular mucins and a loss of this population of sialic acids in dry eye disease in spite of a significant increase in total sialic acids in KCS mucin. PMID- 16311886 TI - Establishment of cells exhibiting mutated glycolipid synthesis from mouse thymus by immortalization with SV-40 virus. AB - Immortalization with simian virus-40 and cloning of immortalized cells from mouse thymus were performed to establish cell lines for characterization of the mode of glycolipid expression in the thymic cells. Among the 25 cell lines obtained, three lines with different morphologies were established, that is, epithelial (IMTH-E), fibroblastic (IMTH-F), and asterisk-like (IMTH-I) cells, and their glycolipids, together with those in the thymus, were determined systematically. The major glycolipids in mouse thymus were the globo- and ganglio-series, both of which, were co-expressed in the three cell lines established. However, the mode of modification of the globo- and ganglio-series was distinct for each cell line. As to the globo-series, the structures with the longest carbohydrate chain for IMTH-E, -F, and -I cells were Gb3Cer, Gb4Cer, and Forssman antigen, respectively, having stepwise shorter carbohydrates at the nonreducing termini. Although the acidic glycolipids in IMTH-E cells comprised GM3 and GM2, and their sulfated isomers, IMTH-F and -I cells expressed GMlb and GDlc for the alpha-pathway, and up to GDI a for the a-pathway of ganglio-series glycolipids. GMlb-GalNAc present in the thymus was not detected in IMTH-F and -I cells, probably due to the lower synthetic activity for the metabolic intermediate Gg4Cer. The results indicate that the immortalization technique is useful for obtaining individual cells having unique glycolipid profiles for analysis of the functional significance and metabolism of glycolipids in the thymus. PMID- 16311887 TI - Convenient and rapid analysis of linkage isomers of fucose-containing oligosaccharides by matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (MALDI-QIT-TOF MS) was used to analyze three pyridylamino (PA) fucosyloligosaccharides isolated from human milk: lacto-N-fucopentaose (LNFP) I [Fucalpha1-2Galbeta1-3GlcNAcbeta1-3Galbeta1-4Glc-PA], LNFP II [Galbeta1 3(Fucalpha1-4)GlcNAcbeta1-3Galbeta1-4Glc-PA], and LNFP III [Galbeta1-4(Fucalpha1 3)GlcNAcbeta1-3Galbeta1-4Glc-PA]. These oligosaccharides are linkage isomers. MALDI-QIT-TOF MS provides MS(n) spectra, which we used to characterize these PA oligosaccharides. MS/MS/MS analysis of the non-reducing end tri-saccharide ions generated by MS/MS was able to distinguish these oligosaccharide isomers. The MALDI-QIT-TOF MS is a very convenient and rapid method, therefore, it would be useful for high throughput structural analyses of various types of pyridylaminated oligosaccharide isomers. PMID- 16311888 TI - Relative amounts of sialic acid and fucose of amniotic fluid glycoconjugates in relation to pregnancy age. AB - The present knowledge concerning the glycan structures and role of glycoconjugates derived from amniotic fluid is fragmentary and mainly focuses on the individual glycoproteins. The question has arisen as whether the general glycosylation pattern of amniotic fluid glycoconjugates can change with the progression of a normal pregnancy. In the present work we have described the dynamic, quantitative alterations in relative amounts of sialic acid and fucose linked by a variety of anomeric linkages to subterminal oligosaccharide structures of amniotic fluid glycoconjugates in relation to pregnancy age. The analysis was performed in the following groups of amniotic fluids derived from normal pregnancy by lectin dotting method: "2nd trimester" (14-19 weeks), "3rd trimester" (29-37 weeks), "perinatal period" (38-40 weeks) , "delivery at term" (39-41 weeks) and "post date pregnancy" (41-43 weeks). In the "3rd trimester" the amniotic fluid glycoconjugates contained higher relative amounts of glycans terminated by alpha2-6-linked sialic acid (p < 0.00002) and by alpha1-6 innermost fucose (p < 0.000001) than those in the 2nd trimester. In contrast, they showed the lower relative amount of fucose linked alpha1-3 (p < 0.02). At the perinatal period the relative amount of alpha2-6-linked sialic acid increased (p < 0.03), and it then decreased during delivery (p < 0.02) to the level found in the "3rd trimester" group. In the post date pregnancy all parameters studied increased. The sialyl- and fucosyl-glycotopes of the amniotic fluid glycoconjugates may play an critical role in growth and tissue remodeling of the foetus, as well as may might reflect maturation of a foetus. Additionally, a determination of the glycotope expressions might be helpful in prenatal diagnosis as predictor factors for well being of mother and child. PMID- 16311889 TI - Biological evaluation of a series of 2-acetamido-2-deoxy-D-glucose analogs towards cellular glycosaminoglycan and protein synthesis in vitro. AB - Using primary hepatocytes in culture, various 2-acetamido-2-deoxy-D-glucose (GlcNAc) analogs were examined for their effects on the incorporation of D [3H]glucosamine, [35S]sulfate, and L-[14C]leucine into cellular glycoconjugates. A series of acetylated GlcNAc analogs, namely methyl 2-acetamido-3,4,6-tri-O acetyl-2-deoxy-alpha-(3) and beta-D-glucopyranoside (4) and 2-acetamido-1,3,4,6 tetra-O-acetyl-2-deoxy-D-glucopyranose (5), exhibited a concentration-dependent reduction of D-[3H]glucosamine, but not of [35S]sulfate incorporation into isolated glycosaminoglycans (GAGs), without affecting L-[14C]leucine incorporation into total protein synthesis. These results suggest that analogs 3 5 exhibit an inhibitory effect on D-[3H]glucosamine incorporation into isolated GAGs by diluting the specific activity of cellular D-[3H]glucosamine and by competing for the same metabolic pathways. In the case of the corresponding series of 4-deoxy-GlcNAc analogs, namely methyl 2-acetamido-3,6-di-O-acetyl-2,4 dideoxy-alpha-(6) and beta-D-xylo-hexopyranoside (7) and 2-acetamido-1,3,6-tri-O acetyl-2,4-dideoxy-D-xylo-hexopyranose (8), compound 8 at 1.0 mM exhibited the greatest reduction of D-[3H]glucosamine and [35S]sulfate incorporation into isolated GAGs, namely to approximately 7% of controls, and a moderate inhibition of total protein synthesis, namely to 60% of controls. Exogenous uridine was able to restore the inhibition of total protein synthesis by compound 8 at 1.0 mM. Isolated GAGs from cultures treated with compound 8 were shown to be smaller in size (approximately 40 kDa) than for control cultures (approximately 77 kDa). These results suggest that the inhibitory effects of compound 8 on cellular GAG synthesis may be mediated by the incorporation of a 4-deoxy moiety into GAGs resulting in premature chain termination and/or by its serving as an enzymatic inhibitor of the normal sugar metabolites. The inhibition of total protein synthesis from cultures treated with compound 8 suggests a uridine trapping mechanism which would result in the depletion of UTP pools and cause the inhibition of total protein synthesis. A 1-deoxy-GlcNAc analog, namely 2 acetamido-3,4,6-tri-O-acetyl-1,5-anhydro-2-deoxy-D-glucitol (9), also exhibited a reduction in both D-[3H]glucosamine and [35S]sulfate incorporation into isolated GAGs by 19 and 57%, of the control cells, respectively, at 1.0 mM without affecting total protein synthesis. The inability of compound 9 to form a UDP sugar and, hence, be incorporated into GAGs presents another metabolic route for the inhibition of cellular GAG synthesis. Potential metabolic routes for each analog's effects are presented. PMID- 16311890 TI - Analysis of sugar chain-binding specificity of tomato lectin using lectin blot: recognition of high mannose-type N-glycans produced by plants and yeast. AB - The sugar chain-binding specificity of tomato lectin (LEA) against glycoproteins was investigated qualitatively using lectin blot analysis. Glycoproteins containing tri- and tetra-antennary complex-type N-glycans were stained with LEA. Unexpectedly, glycoproteins containing high mannose-type N-glycans and a horseradish peroxidase were stained with LEA. LEA blot analysis of the glycoproteins accompanied by treatment with exoglycosidase revealed that the binding site of LEA for the complex-type N-glycans was the N-acetyllactosaminyl side chains, whereas the proximal chitobiose core appeared to be the binding site of LEA for high mannose-type N-glycans. Despite these results, the glycoproteins did not inhibit the hemagglutinating activity of LEA. Among the chitin-binding lectins compared, potato tuber lectin showed specificity similar to LEA on lectin blot analysis, while Datura stramonium lectin and wheat germ agglutinin (WGA) did not interact with glycoproteins containing high mannose-type N-glycans, except that RNase B was stained by WGA. Based on these observations, LEA blot analysis was applied to sugar chain analysis of tomato glycoproteins. The most abundant LEA-reactive glycoprotein was purified from the exocarp of ripe tomato fruits, and was identified as the tomato anionic peroxidase1 (TAP1). These results suggest that LEA interacts with glycoproteins produced by tomatoes, which participate in biological activities in tomato plants. PMID- 16311891 TI - Unexpected clinicopathological phenotype linked to small elongation of CAG repeat in SCA1 gene. PMID- 16311892 TI - Genetic interaction between two apolipoprotein E receptors increases Alzheimer's disease risk. PMID- 16311893 TI - Recurrent neuropathy associated with Ehlers-Danlos syndrome. PMID- 16311894 TI - CNS demyelination during anti-tumor necrosis factor alpha therapy. PMID- 16311895 TI - MuSK-antibody positive pure ocular myasthenia gravis. PMID- 16311896 TI - Correlates of subjective and functional outcomes in outpatient clinic attendees with schizophrenia and schizoaffective disorder. AB - Outcome in schizophrenia is multidimensional and, thus, consists of clinical,humanitarian, rehabilitative and cost domains. Accordingly, recovery is conceptualized as the ability to function in the community, socially and vocationally, as well as being relatively free of disease-related psychopathology. The present cross-sectional study examined the relationship of premorbid functioning, psychopathology, insight, attitudes toward medication and side-effects, as well as sociodemographic factors with treatment outcomes in terms of quality of life (QOL) and psychosocial functioning among 60 regular attendees of a specialized schizophrenia outpatient clinic. Both insight into the illness as well as attitudes toward treatment indicated satisfactory compliance. Premorbid school and social functioning were positively correlated with actual employment status, and premorbid social functioning was further positively correlated with QOL and global functioning. Attitudes toward treatment were positively associated with global QOL, and with the patients' living situation. Both positive and negative symptoms as well as asthenia were negatively associated with QOL, while cognitive symptoms negatively influenced occupational functioning. Older patients lived independently and/or in a partnership more often, but had a lesser likelihood of competitive employment. Our observations suggest that subjective and functional outcomes in highly compliant patients are mainly predicted by psychopathological symptoms and unchangeable sociodemographic variables. PMID- 16311897 TI - Regional cerebral blood flow change in a case of Alzheimer's disease with musical hallucinations. AB - We examined alteration of regional cerebral blood flow (rCBF) in a case of Alzheimer's disease (AD) patient with musical hallucination. To detect regions related to musical hallucination, single-photon emission computed tomography (SPECT) imaging of the patient and nine sex, age, and cognitive function-matched AD patients without delusions and hallucinations were compared using statistical parametric mapping 99 (SPM99). In comparison with controls, the patient had increased rCBF in left temporal regions and left angular gyrus. This profile could be relevant to the neuroanatomical basis of musical hallucinations. PMID- 16311898 TI - The enduring effects of abuse and related adverse experiences in childhood. A convergence of evidence from neurobiology and epidemiology. AB - BACKGROUND: Childhood maltreatment has been linked to a variety of changes in brain structure and function and stress-responsive neurobiological systems. Epidemiological studies have documented the impact of childhood maltreatment on health and emotional well-being. METHODS: After a brief review of the neurobiology of childhood trauma, we use the Adverse Childhood Experiences (ACE) Study as an epidemiological "case example" of the convergence between epidemiologic and neurobiological evidence of the effects of childhood trauma. The ACE Study included 17,337 adult HMO members and assessed 8 adverse childhood experiences (ACEs) including abuse, witnessing domestic violence, and serious household dysfunction. We used the number of ACEs (ACE score) as a measure of cumulative childhood stress and hypothesized a "dose-response" relationship of the ACE score to 18 selected outcomes and to the total number of these outcomes (comorbidity). RESULTS: Based upon logistic regression analysis, the risk of every outcome in the affective, somatic, substance abuse, memory, sexual,and aggression-related domains increased in a graded fashion as the ACE score increased (P <0.001). The mean number of comorbid outcomes tripled across the range of the ACE score. CONCLUSIONS: The graded relationship of the ACE score to 18 different outcomes in multiple domains theoretically parallels the cumulative exposure of the developing brain to the stress response with resulting impairment in multiple brain structures and functions. PMID- 16311900 TI - Modulation of antioxidant enzyme activities, platelet aggregation and serum prostaglandins in rats fed spray-dried milk containing n-3 fatty acid. AB - Spray-dried milk enriched with n-3 fatty acids from linseed oil or fish oil were fed to rats to study its influence on liver lipid peroxides, hepatic antioxidant enzyme activities, serum prostaglandins and platelet aggregation. Significant level of alpha linolenic acid, eicosapentaenoic acid and docosahexaenoic acid were accumulated at the expense of arachidonic acid in the liver of rats fed n-3 fatty acid enriched formulation. The linseed oil and fish oil enriched formulation fed group had 44 and 112% higher level of lipid peroxides in liver homogenate compared to control rats fed groundnut oil enriched formulation. Catalase activity in liver homogenate was increased by 37 and 183% respectively in linseed oil and fish oil formulation fed rats. The glutathione peroxidase activity decreased to an extent of 25-36% and glutathione transferase activity increased to an extent of 34-39% in rats fed n-3 fatty acids enriched formulation. Feeding n-3 fatty acid enriched formulation significantly elevated the n-3 fatty acids in platelets and increased the lipid peroxide level to an extent of 4.2-4.5 fold compared to control. The serum thromboxane B2 level was decreased by 35 and 42% respectively in linseed oil and fish oil enriched formulation fed rats, whereas, 6-keto- prostaglandin F1alpha level was decreased by 17 and 23% respectively in linseed oil and fish oil enriched formulation fed rats. The extent and rate of platelet aggregation was decreased significantly in n-3 fatty acids enriched formulation fed rats. This indicated that n-3 fatty acids enriched formulation beneficially reduces platelet aggregation and also enhances the activities of hepatic antioxidant enzymes such as catalase and glutathione transferase. PMID- 16311901 TI - Pyridoxal phosphate inhibits the group I intron splicing. AB - The coenzyme pyridoxal phosphate and its analogs were tested for inhibition of the self-splicing of primary transcripts of the phage T4 thymidylate synthase gene (td). Of all compounds examined, the pyridoxal phosphate was the most potent inhibitor and the order of inhibitory efficiency for compounds tested was as follows: pyridoxal phosphate > pyridoxal > pyridoxine > pyridoxamine > pyridoxic acid. Increasing Mg2+ concentration up to 14 mM overcame the suppression of self splicing by pyridoxal phosphate up to 95% of the level of normal splicing, implying its interference with effective catalysis of Mg2+. The kinetic analysis demonstrated that pyridoxal phosphate acts as a mixed type noncompetitive inhibitor for the td intron RNA with a K(i) of 11.8 mM. The specificity of the splicing inhibition by pyridoxal phosphate is predominantly due to increases in K(m) and decreases in V(max) values. PMID- 16311902 TI - High dietary vitamin C affects growth, non-specific immune responses and disease resistance in Asian catfish, Clarias batrachus. AB - Studies were conducted to determine the immunomodulatory effects of high dietary ascorbic acid (vitamin C) on growth, serum concentration, non-specific immune response and disease resistance of a commercially important Asian catfish, Clarias batrachus. Four practical diets were formulated to contain 0, 500, 1000 and 2000 mg ascorbic acid (AA) equivalent/kg diet, supplied as L-ascorbyl-2 polyphosphate (LAPP) and were fed for 2, 4, 6 and 8 weeks. Each diet was fed to triplicate groups of catfish with initial body weight of 15.47+/- 0.59 g. After 2, 4, 6 and 8 weeks, growth, serum concentration of AA, oxidative respiratory burst, lysozyme and natural hemolytic complement activities, myeloperoxidase (MPO) content and natural haemagglutination titre were measured. Ten numbers of fish in duplicate were challenged with Aeromonas hydrophila to measure the level of protection against aeromoniasis at each one of the assayed times. The results showed that AA concentration in serum correlated positively with those in the diets and reached its saturation level after the time period directly proportional to the increase in dose level. Fish fed AA-supplemented diets showed significantly (p < 0.05) higher specific growth rate after 2 weeks of feeding. The superoxide production was enhanced after 8 weeks of feeding fish at a supplemented dose level of 2000 mg/kg. Similarly, MPO content, haemagglutination titre and alternative complement activity in serum enhanced with the increase of dietary AA levels at different duration of feeding. The lysozyme activity was not affected by the dietary AA treatment. On the other hand, feeding of AA at all concentrations significantly increased percent survival against A. hydrophila challenge after 4 weeks compared to control. The non-specific immune parameters as well as percent survival were enhanced as a result of high AA supply particularly at 500 mg/kg diet, although the increase was not maintained but returned to the initial levels after 4 weeks. These results support the possible use of AA as an immunostimulant at a dose of 500 mg/kg diet for a period of 4 weeks in catfish farming. PMID- 16311903 TI - Agonist-induced phosphorylation and desensitization of the P2Y2 nucleotide receptor. AB - Purification of HA-tagged P2Y2 receptors from transfected human 1321N1 astrocytoma cells yielded a protein with a molecular size determined by SDS-PAGE to be in the range of 57-76 kDa, which is typical of membrane glycoproteins with heterogeneous complex glycosylation. The protein phosphatase inhibitor, okadaic acid, attenuated the recovery of receptor activity from the agonist-induced desensitized state, suggesting a role for P2Y2 receptor phosphorylation in desensitization. Isolation of HA-tagged P2Y2 nucleotide receptors from metabolically [32P]-labelled cells indicated a (3.8 +/- 0.2)-fold increase in the [32P]-content of the receptor after 15 min of treatment with 100 microM UTP, as compared to immunoprecipitated receptors from untreated control cells. Receptor sequestration studies indicated that approximately 40% of the surface receptors were internalized after a 15-min stimulation with 100 microM UTP. Point mutation of three potential GRK and PKC phosphorylation sites in the third intracellular loop and C-terminal tail of the P2Y2 receptor (namely, S243A, T344A, and S356A) extinguished agonist-induced receptor phosphorylation, caused a marked reduction in the efficacy of UTP to desensitize P2Y2 receptor signalling to intracellular calcium mobilization, and impaired agonist-induced receptor internalization. Activation of PKC isoforms with phorbol 12-myristate 13-acetate that caused heterologous receptor desensitization did not increase the level of P2Y2 receptor phosphorylation. Our results indicate a role for receptor phosphorylation by phorbol-insensitive protein kinases in agonist-induced desensitization of the P2Y2 nucleotide receptor. PMID- 16311904 TI - Versican is induced in infiltrating monocytes in myocardial infarction. AB - Versican, a large chondroitin sulfate proteoglycan, plays a role in conditions such as wound healing and tissue remodelling. To test the hypothesis that versican expression is transiently upregulated and plays a role in the infarcted heart, we examined its expression in a rat model of myocardial infarction. Northern blot analysis demonstrated increased expression of versican mRNA. Quantitative real-time RT-PCR analysis revealed that versican mRNA began to increase as early as 6 h and reached its maximal level 2 days after coronary artery ligation. Versican mRNA then gradually decreased, while the mRNA of decorin, another small proteoglycan, increased thereafter. Versican mRNA was localized in monocytes, as indicated by CD68-positive staining, around the infarct tissue. The induction of versican mRNA was accelerated by ischemia/reperfusion (I/R), which was characterized by massive cell infiltration and enhanced inflammatory response. To examine the alteration of versican expression in monocytes/macrophages, we isolated human peripheral blood mononuclear cells and stimulated them with granulocyte/macrophage colony stimulating factor (GM-CSF). Stimulation of mononuclear cells with GM-CSF increased the expression of versican mRNA as well as cytokine induction. The production of versican by monocytes in the infarct area represents a novel finding of the expression of an extracellular matrix gene by monocytes in the infarcted heart. We suggest that upregulation of versican in the infarcted myocardium may have a role in the inflammatory reaction, which mediates subsequent chemotaxis in the infarcted heart. PMID- 16311905 TI - Protection of cellular DNA from gamma-radiation-induced damages and enhancement in DNA repair by troxerutin. AB - The effect of troxerutin on gamma-radiation-induced DNA strand breaks in different tissues of mice in vivo and formations of the micronuclei were studied in human peripheral blood lymphocytes ex vivo and mice blood reticulocytes in vivo. Treatments with 1 mM troxerutin significantly inhibited the micronuclei induction in the human lymphocytes. Troxerutin protected the human peripheral blood leucocytes from radiation-induced DNA strand breaks in a concentration dependent manner under ex vivo condition of irradiation (2 Gy). Intraperitoneal administration of troxerutin (175 mg/kg body weight) to mice before and after whole body radiation exposure inhibited micronuclei formation in blood reticulocytes significantly. The administration of different doses (75, 125 and 175 mg/kg body weight) of troxerutin 1 h prior to 4 Gy gamma-radiation exposure showed dose-dependent decrease in the yield of DNA strand breaks in murine blood leucocytes and bone marrow cells. The dose-dependent protection was more pronounced in bone marrow cells than in blood leucocytes. Administration of 175 mg/kg body weight of the drug (i.p.) 1 h prior or immediately after whole body irradiation of mice showed that the decrease in strand breaks depended on the post-irradiation interval at which the analysis was done. The observed time dependent decrease in the DNA strand breaks could be attributed to enhanced DNA repair in troxerutin administered animals. Thus in addition to anti-erythrocytic, anti-thrombic, fibrinolytic and oedema-protective rheological activity, troxerutin offers protection against gamma-radiation-induced micronuclei formation and DNA strand breaks and enhances repair of radiation-induced DNA strand breaks. PMID- 16311906 TI - Effect of chronic endothelin blockade on PKC isoform distribution in mesenteric arteries from diabetic rats. AB - Hemodynamic changes, including increased vasoconstriction and reduced blood flow have been detected in both human diabetic patients and in animal models of diabetes. We previously demonstrated that the endothelin (ET) system was upregulated and involved in mediating the exaggerated vasoconstrictor responses in superior mesenteric artery (SMA) from diabetic rats. Chronic treatment of diabetic rats with the dual endothelin receptor antagonist, bosentan abolished the enhanced contractile responses in diabetic SMA. The biological actions of ET 1 have been shown to be coupled to the hydrolysis of phosphotidylinositol 4,5 biphosphate and phosphotidylcholine and the subsequent production of diacylglycerol (DAG). DAG is an activator of the classical and novel isoforms of PKC. Increases in PKC activity, associated with translocation of specific PKC isoforms from the cytosol to the membrane, have been implicated in the vasoconstrictor effect of ET-1. The goal of the present study was to determine whether chronic treatment of diabetic rats with bosentan influences the activation of specific PKC isoforms in SMA from diabetic rats. Elevated levels of PKCbeta2 in both the cytosol and membrane fractions and PKCepsilon in the membrane fraction were detected in SMA from diabetic rats. However, neither the levels nor the distribution between the cytosol and membrane fractions of any of these PKC isoforms were affected by the treatment of the diabetic rats with bosentan. These observations indicate that bosentan improves vascular reactivity in STZ-diabetic rats by mechanisms other than correction of increased activities of PKC isoforms. PMID- 16311907 TI - Sensitivity of mitochondria isolated from liver and kidney of rat and bovine to lipid peroxidation: a comparative study of light emission and fatty acid profiles. AB - Much work has been carried out on non-enzymatic-induced lipid peroxidation of mitochondria obtained from different tissues of monogastric animals, but little information is available about this process in poligastric animals. Studies were carried out to determine the sensitivity of mitochondria isolated from liver and kidney of rat and bovine to lipid peroxidation (ascorbate-Fe2+ dependent) by comparison of light emission and fatty acid profiles. Mitochondria from both species were susceptible to lipid peroxidation. Measurements of chemiluminescence indicate that the lipid peroxidation process was more effective in mitochondria from rat liver than in the organelle obtained from bovine, whereas changes were not observed in mitochondria from rat and bovine kidney. The fatty acid composition of total lipids isolated from liver and kidney mitochondria of both species was substantially modified when subjected to non-enzymatic lipid peroxidation with a decrease of arachidonic and docosahexaenoic acids. The polyunsaturated fatty acid (PUFA) composition was higher in mitochondria obtained from rat liver (43.11+/- 4.16) than in bovine (15.78 +/- 0.76). As a consequence, the unsaturation index (UI), was higher in mitochondria of rat liver than in bovine. Nevertheless, the PUFA composition of kidney mitochondria from both species was similar; therefore, statistically significant differences in the UI were not observed. The results suggest that mainly the PUFAs present in hepatic and kidney mitochondria were sensitive to oxidative damage. The lipid peroxidation process was more effective in rat liver mitochondria than in bovine. PMID- 16311908 TI - Age-associated deficit of mitochondrial oxidative phosphorylation in skeletal muscle: role of carnitine and lipoic acid. AB - Mitochondrial damage has implicated a major contributor for ageing process. In the present study, we measured mitochondrial membrane swelling, mitochondrial respiration (state 3 and 4) by using oxygen electrode in skeletal muscle of young (3-4 months old) and aged rats (above 24 months old) with supplementation of L: carnitine and DL: -alpha-lipoic acid. Our results shows that the mitochondrial membrane swelling and state 4 respiration were increased more in skeletal muscle mitochondria of aged rats than in young control rats, whereas the state 3 respiration, respiratory control ratio (RCR) and ADP:O ratio decreased more in aged rats than in young rats. After supplementation of carnitine and lipoic acid to aged rats for 30 days, the state 3 respiration and RCR were increased, whereas the state 4 and mitochondrial membrane swelling were decreased to near normal rats. From our results, we conclude that combined supplementation of carnitine and lipoic acids to aged rats increases the skeletal muscle mitochondrial respiration, thereby increasing the level of ATP. PMID- 16311909 TI - A 96-well automated method to study inhibitors of human sodium-dependent D glucose transport. AB - The sodium-dependent D-glucose transporter (SGLT) family is involved in glucose uptake via intestinal absorption (SGLT1) or renal reabsorption (SGLT1 and SGLT2). Current methods for the screening of inhibitors of SGLT transporters are complex, expensive and very labor intensive, and have not been applied to human SGLT transporters. The purpose of the present study was to develop an alternative 96 well automated method to study the activity of human SGLT1 and SGLT2. Chinese hamster ovary (CHO) Flp-In cells were stably transfected with pcDNA5-SGLT1 or pcDNA5-SGLT2 plasmid and maintained in hygromycin-selection Ham's F12 culture medium until hygromycin-resistant clones were developed. SGLT1 and SGLT2 gene expression was evaluated by relative real-time reverse transcription-polymerase chain reaction (RT-PCR) quantification, Western blotting, and immunocytochemical analysis. The clones with higher expression of SGLT1 and SGLT2 were used for transport studies using [14C]-methyl-alpha-D-glucopyranoside ([14C]AMG). The advantage of using the 96-well format is the low amount of radioactive compounds and inhibitory substances required, and its ability to establish reproducibility because repetition into the assay. This method represents an initial approach in the development of transport-based high-throughput screening in the search for inhibitors of glucose transport. The proposed method can easily be performed to yield quantitative data regarding key aspects of glucose membrane transport and kinetic studies of potential inhibitors of human SGLT1 and SGLT2. PMID- 16311910 TI - HIF-1alpha involvement in low temperature and anoxia survival by a freeze tolerant insect. AB - Winter survival for many insect species relies on the ability to endure the freezing of extracellular body fluids. Because freezing impedes oxygen delivery to tissues, one component of natural freeze tolerance is a well-developed anoxia/ischemia resistance. The present study explores the responses of the hypoxia-inducible factor-1alpha (HIF-1alpha) to cold, freezing and anoxia exposures in the freeze tolerant goldenrod gall fly larva, Eurosta solidaginis. Reverse transcription-PCR was used to quantify hif-1alpha transcript levels; transcripts were significantly elevated by approximately 70% in chilled (3 ( composite function)C), frozen (-16 ( composite function)C) and thawed (returned to 3 ( composite function)C) insects, compared with 15 ( composite function)C controls. Transcripts also rose by approximately 3-fold in insects given anoxia exposure under a nitrogen gas atmosphere. Cold and freezing exposure also elevated HIF-1alpha protein content in the larvae and HIF-1alpha levels increased over the winter months in insects sampled from an outdoor population; levels peaked in February at 2.1-fold higher than in September. A partial sequence of HIF-1alpha that covers the bHLH and PAS domains of the protein was obtained from E. solidaginis and sequence analysis revealed that this segment shared 62% identity overall with Drosophila melanogaster HIF-1alpha and higher percent identities within specific domains: 76% within the bHLH domain and 70% within the PAS domain. The data provide the first documentation of a potential role for HIF 1 in regulating the expression of genes that can aid freezing survival in a cold hardy animal. PMID- 16311911 TI - Role of MMP-2 in PKCdelta-mediated inhibition of Na+ dependent Ca2+ uptake in microsomes of pulmonary smooth muscle: involvement of a pertussis toxin sensitive protein. AB - Treatment of bovine pulmonary artery smooth muscle with the O2 *- generating system hypoxanthine plus xanthine oxidase stimulated MMP-2 activity and PKC activity; and inhibited Na+ dependent Ca2+ uptake in the microsomes. Pretreatment of the smooth muscle with SOD (the O2 *- scavenger) and TIMP-2 (MMP-2 inhibitor) prevented the increase in MMP-2 activity and PKC activity, and reversed the inhibition of Na+ dependent Ca2+ uptake in the microsomes. Pretreatment with calphostin C (a general PKC inhibitor) and rottlerin (a PKCdelta inhibitor) prevented the increase in PKC activity and reversed O2 *- caused inhibition of Na+ dependent Ca2+ uptake without causing any change in MMP-2 activity in the microsomes of the smooth muscle. Treatment of the smooth muscle with the O2 *- generating system revealed, respectively, 36 kDa RACK-1 and 78 kDa PKCdelta immunoreactive protein profile along with an additional 38 kDa immunoreactive fragment in the microsomes. The 38 kDa band appeared to be the proteolytic fragment of the 78 kDa PKCdelta since pretreatment with TIMP-2 abolished the increase in the 38 kDa immunoreactive fragment. Co-immunoprecipitation of PKCdelta and RACK-1 demonstrated O2 *- dependent increase in PKCdelta-RACK-1 interaction in the microsomes. Immunoblot assay elicited an immunoreactive band of 41 kDa G(i)alpha in the microsomes. Treatment of the smooth muscle tissue with the O2 *- generating system causes phosphorylation of G(i)alpha in the microsomes and pretreatment with TIMP-2 and rottlerin prevented the phosphorylation. Pretreatment of the smooth muscle tissue with pertussis toxin reversed O2 *- caused inhibition of Na+ dependent Ca2+ uptake without affecting the protease activity and PKC activity in the microsomes. We suggest the existence of a pertussis toxin sensitive G protein mediated mechanism for inhibition of Na+ dependent Ca2+ uptake in microsomes of bovine pulmonary artery smooth muscle under O2 *- triggered condition, which is regulated by PKCdelta dependent phosphorylation and sensitive to TIMP-2 for its inhibition. PMID- 16311912 TI - The effect of time of day and exercise on platelet functions and platelet neutrophil aggregates in healthy male subjects. AB - Platelet activation state changes by exercise. The effect of exercise time on platelet activation state and formation of platelet-neutrophil aggregates are not known yet. In this study the effect of exercise and time of day were examined on platelet activity with platelet-neutrophil aggregates. Ten moderately active males aged 27+/- 1.63 (mean+/-S.D.) years completed sub-maximal (70% VO(2max)) exercise trials for 30 min. Blood pressure (BP) was recorded. Venous blood samples were obtained at rest, immediately post-exercise and after 30 min of recovery. Whole blood was analysed for haematocrit (Hct), haemoglobin (Hb), platelet count (PC), mean platelet count (MPV) and platelet aggregation (PA). Platelet-neutrophil aggregates and beta-thromboglobulin (beta-TG) levels were assayed. Platelet count showed significant increase after morning exercise ((236+/- 32)x10(9) l(-1) versus (202+/- 34)x10(9) l(-1) baseline, p < 0.05). Exercise resulted in significantly lower MPV after the evening exercise (9.16+/- 0.5 fl versus 9.65+/- 0.36 fl, p < 0.05). Platelet aggregation by adenosine diphosphate (ADP) decreased after morning exercise and the recovery aggregation levels were significantly different at two different times of the day (68+/- 20% a.m. versus 80+/- 12% p.m., p < 0.05). It was also showed that platelet neutrophil aggregates increased significantly from baseline after both exercises. Exercise-induced platelet-neutrophil aggregates were higher in the evening (10.7+/- 1.3% p.m. versus 6.4+/- 1.8% a.m., p < 0.0001). It is therefore concluded that besides platelet-platelet aggregation, exercise can cause platelet neutrophil aggregates. In addition, time of day has an effect on platelet activation related events. Circadian variations of physiological parameters may have an effect on thrombus formation by platelet activation. PMID- 16311913 TI - A herbal medicine for the treatment of lung cancer. AB - Lung cancer is the leading cause of cancer-related deaths throughout the world. Extracts of medicinal plants are believed to contain different chemopreventive or chemotherapeutic compounds. In this study, we determined the anti-cancer property of one of the traditional Indian medicine Rasagenthi Lehyam (RL) for the treatment of lung cancer. Two lung cancer cell lines (A-549 and H-460) and one normal bronchial epithelial (BEAS-2B) cell line were used to test the chemotherapeutic effect of RL. Out of five fractions of RL, chloroform fraction of RL (cRL) demonstrated a significant inhibition of cell proliferation and induction of apoptosis in A-549 and H-460 cells but not in normal BEAS-2B cells. The cRL fraction up-regulated the pro-apoptotic genes p53 and Bax and induced caspase-3 activation, and down-regulated the pro-survival gene Bcl-2 in both the lung cancer cell lines. Also, nuclear export of p53 was seen in cRL-treated lung cancer cells. In addition, cRL induced G2/M arrest of cell cycle and enhanced the radio-sensitivity of both the lung cancer cell lines. This study suggests that cRL may prove to be a potent anti-cancer agent that may be used for the treatment of lung cancer. However, further studies are required to bring cRL into the mainstream of medicine in the treatment of lung cancer. PMID- 16311914 TI - The effects of vitamin C supplementation on oxidative stress and antioxidant content in the brains of chronically exercised rats. AB - The purpose of this study was to ascertain whether vitamin C supplementation during chronic exercise training alters rat brain antioxidant content. Female Wistar albino rats were exercised on a treadmill for 30 min/day for 6.5 weeks and were administered daily intraperitoneal injections of vitamin C (20 mg/kg). After the training period, chronically exercised rats showed no significant changes in total brain thiobarbituric acid reactive substances (TBARS) levels. In contrast, rats supplemented with vitamin C during the training period showed significantly elevated brain TBARS levels. If such results were extrapolated to man, where vitamin supplementation is a common practice, this would indicate that vitamin C supplementation may not protect brain tissue against exercise-induced oxidative damage, in such circumstances, this water-soluble antioxidant behaves as a pro oxidant. PMID- 16311915 TI - Cadmium induced MTs synthesis via oxidative stress in yeast Saccharomyces cerevisiae. AB - Yeast, Saccharomyces cerevisiae, exposed to CdCl2 for 17 h was analysed with reference to survival, MTs and oxidative stress biomarkers. An enhanced accumulation of MDA and the increased activities of SOD and GPx in the Cd-treated yeasts under aerobic condition indicated CdCl2-caused oxidative stress in S. cerevisiae. MTs were significantly induced by CdCl2 under aerobic condition and the induced MTs contents were positively correlated with the accumulation of MDA in this study. However, MTs induction can be prominently inhibited by coincubation with NAC or anaerobic culture via eliminating ROS. This oxidative stress reduction was reflected by the decreases in MDA level and SOD and GPx activities. The results suggest that MTs inductive activity of cadmium in yeast cells was mediated by oxidative stress. In addition, increase of MTs contents was observed in cells untreated with CdCl2 under anaerobic conditions or coincubation with NAC, suggesting MTs are also susceptive to reductive stress. PMID- 16311916 TI - Expression of inducible nitric oxide synthase in human cutaneous leishmaniasis. AB - Cutaneous leishmaniasis (CL) is an infectious disease caused by Leishmania parasite. The expression of inducible nitric oxide synthase (iNOS) and generation of nitric oxide in response to IFN-gamma and TNF-alpha is important in control of infection. The aim of the study was to determine the expression of iNOS in the lesions of Leishmania tropica, and whether there was a correlation between the level of expression and the duration of the disease. Punch biopsy was performed from patients (n = 29) and iNOS immunohistochemical staining was applied. Expression of iNOS protein was detected 82.8% of patients. There was a strong expression with the duration of the disease less than 6 months (p < 0.002). These findings demonstrate that iNOS has a role in L. tropica especially during the early stages of the infection. PMID- 16311917 TI - Effects of dexfenfluramine on serotonin levels of mice ileum, contractility, glutathione and malondialdehyde level. AB - Dexfenfluramine is one of the anorectic drugs that suppresses food intake which acts via inhibition of reuptake of serotonin into brain terminal. Gastrointestinal tract is the main source of peripheral serotonin which is involved in the regulation of gastrointestinal motility. During the use of anorectic drugs, the antioxidant defence is affected especially by reactive oxygen species. The purpose of this study to search: The effect of dexfenfluramine on serotonin levels of ileum and the effect of dexfenfluramine on ileal contractility and oxidative stress. MATERIALS AND METHODS: Twenty-two adult male Swiss-albino mice were divided two groups (1) Control, (2) Dexfenfluramine treated (i.p. twice a day 0.2 mg kg(-1) in 0.2 ml saline solution for 7 days). Animal body weights were recorded at the beginning and at the end of the experimental period. Ileum tissues contractile responses to different concentrations of KCl and acethycholine were recorded on polygraph. In the meantime ileal tissue malondialdehyde, a product of lipid peroxidation, and glutathione, endogenous antioxidant levels were assessed by spectrophotometric methods. Ileal tissue serotonin level determined by immunohistochemical method. Body weights decrease and ileal contractile response of acethycholine increased significantly by dexfenfluramine treatment. Meanwhile, ileum glutathione levels decreased and malondialdehyde levels increased in dexfenfluramine treated group. Immunohistochemical detection showed that ileal serotonin levels increased by dexfenfluramine treatments. As a conclusion, there is a relationship between increased ileal contractility and oxidant status in dexfenfluramine treated animals. These effects can be related by increased serotonin levels which is induced by dexfenfluramine in ileum. PMID- 16311918 TI - Subchronic exposure to high-dose ACE-inhibitor moexipril induces catalase activity in rat liver. AB - The long-term clinical effects of ACE-inhibitors have similarities with those of both fibrates and glitazones, activators of peroxisome proliferator activator receptor (PPAR) alpha and gamma, respectively. The antioxidant enzyme catalase, a heme protein that degrades hydrogen peroxide, is found at high concentrations in peroxisomes. Catalase activity is one of the recognized surrogate markers indicative of PPAR activation in the rat liver. The purpose of the study was to establish the effect of moexipril on catalase activity and to compare it with the effect of both saline controls and that of the known PPAR agonist clofibrate (positive control). Three groups of seven rats were used. All substances were applied i.p. daily for 5 days, followed by a 2-day break. The cycle was repeated eight times. After the final cycle (day 56) the animals were sacrificed and liver tissue collected. The number of catalase positive cells in both moexipril group (95% CI 57-61) and clofibrate group (95% CI 72-80) is higher than in controls (95% CI 3-16) (p < or = 0.01). The number of catalase positive cells in the clofibrate group is higher than in the moexipril group (p < or = 0.01). High-dose subchronic exposure to the ACE-inhibitor moexipril induces catalase activity in the rat liver to an extent comparable to fibrates. We suggest that some of the long-term advantages of ACE inhibitor use - beyond mere BP lowering - might be due to a PPAR mediated effect. PMID- 16311919 TI - The inhibitory effect of sodium selenite on N-nitrosodiethylamine-induced and phenobarbital promoted liver tumourigenesis in rats based on the modulation of polyamine levels. AB - In the present study, we have evaluated the effects of dietary selenite (Se) on polyamine levels and its influence on N-nitrosodiethylamine (DEN) initiated and Phenobarbital (PB) promoted in rat liver carcinogenesis. Dietary selenite at a concentration of 4 ppm (through drinking water) was administered in rats either before initiation (4 weeks), or during promotion (16 weeks) and entire experimental period (20 weeks). Male Wistar strain of albino rats was treated with single intra peritoneal dose of DEN (200 mg kg(-1) body weight), after 2 weeks the carcinogenic effect was promoted by PB (0.05%; through diet). Alpha fetoprotein (AFP) was investigated after the 20th-week of experimental period. Selenite-treated animals markedly reduced the AFP during the time of pre-selenite [before initiation (4 weeks)] and entire experimental period (20 weeks), administration rather than the promotion period. This infers that anticancer property of selenite depends on the stage of carcinogenesis, rather than duration of treatment. Evaluation of polyamine levels in hepatoma and surrounding liver tissue showed significant difference in the selenite-treated groups compared with pair-fed control groups. Furthermore, histopathological examination showing remarkable difference between control and treated groups. These results demonstrate that selenite can modulate the development of DEN-induced and PB promoted rat liver carcinogenesis through a polyamine-dependent mechanism. PMID- 16311920 TI - Interaction of stannous chloride leads to alteration in DNA, triphosphate nucleotides and isolated bases. AB - Stannous chloride (SnCl2) is a reducing chemical agent used in several man-made products. SnCl2 can generate reactive oxygen species (ROS); therefore, studies have been carried out in order to better understand its damaging action in biological systems. In this work, calf thymus DNA, triphosphate nucleotides and isolated bases were incubated with SnCl2 and the results were analyzed through UV spectrophotometry. The presence of stannous ions altered the absorption spectra of all three isolates. The amount of stannous ions associated to DNA was measured by atomic absorption spectrophotometry. Data showed that more than 40% of the initial SnCl2 concentration was present in the samples. Our results are in accordance with the damaging potential of this salt and present evidence that stannous ions can complex with DNA, inducing ROS in its vicinity, which may be responsible for the observed lesions. PMID- 16311921 TI - Antagonism between olive oil phenolics and nitric oxide on lymphomonocyte cytosolic calcium. AB - Some biological actions of olive oil phenolics (inhibition of platelet aggregation, decrease of LDL-oxidation, inhibition of bacterial growth and hypertensive action) have been attributed to NOS stimulation in endothelial cells through an increase of cytosolic calcium, notwithstanding the scavenging activity of phenolics on NO and superoxide. In this paper, we determine the concentration of cytosolic calcium in human lymphomonocytes incubated with high concentrations of NO-donors (CysNO) and we evaluate the effects of olive oil phenolics on this parameter. CysNO induces a marked decrease of cytosolic calcium; both olive oil phenolics oppose this action of CysNO. The effects of phenolics and CysNO are independent and additive. PMID- 16311922 TI - Cloning of cDNAs with PDCD2(C) domain and their expressions during apoptosis of HEK293T cells. AB - There are only two isoforms of PDCD2 and MGC13096 containing PDCD2(C) domain in human genome. To study the role of PDCD2_C domain in apoptosis, the cDNAs of two isoforms of PDCD2 and MGC13096 were cloned. The RT-PCR products (AY948416, AY948417) of PDCD2 from RNA of human embryonic kidney 293T (HEK293T) and gastric cancer AGS cell line lost common 99 bp when compared with the sequences of NCBI database (NM_002598, NM_144781). The data of expression of PDCD2 and MGC13096 genes in HEK293T cells which induced to undergo apoptosis by various treatments suggested that there was no significant over-regulation of MGC13096 gene and the over-expression of PDCD2 gene did not occur universally. We searched GEO (Gene Expression Omnibus) about PDCD2 and MGC13096. PDCD2 (NM_002598) was over expressed when endothelial cells treated with leukotriene D4 or natural killer cells were activated by IL-2. Perhaps PDCD2_C domain is not universally associated with apoptosis, the function of PDCD2_C domain needs to be studied further. PMID- 16311923 TI - The protective role of erdosteine on testicular tissue after testicular torsion and detorsion. AB - Testicular torsion and detorsion are important clinical problems for infertile man and oxidative stress may have a role in this clinical situation. The aim of this study was to investigate the protective role of erdosteine, an antioxidant, on unilateral testicular reperfusion injury in rats. The rats were divided into four groups including seven rats in each group: control, torsion, torsion/detorsion and torsion/detorsion+erdosteine. Rats, except the sham operation group, were subjected to left unilateral torsion (720( composite function) rotation in the clockwise direction) without including the epididymis. The experiments were finished after sham operation time for control, 120 min torsion for torsion group and 120 min torsion and 240 min detorsion for torsion/detorsion groups. Bilateral orchiectomy was performed for all groups of rats. The ipsilateral and controlateral testis were divided into two pieces to analyse biochemical parameters and to investigate the light microscopic view. Malondialdehyde level of ipsilateral testis was increased in torsion and torsion/detorsion groups in comparison with the other groups (p < 0.05). Erdosteine treatment ameliorated lipid peroxidation after torsion/detorsion in ipsilateral testis (p < 0.05). Also, xanthine oxidase activity of ipsilateral testis was increased in torsion/detorsion group in comparison with the others (p < 0.05). Nitric oxide (NO) level of ipsilateral testis was higher in all experimental groups than sham operated control group (p < 0.05). Also, NO level of torsion group was increased in comparison with detorsion groups (p < 0.05). Erdosteine treatment caused increased glutathione peroxidase activity in comparison with torsion and torsion/detorsion groups and catalase activity in comparison with the other groups in ipsilateral testis (p < 0.05). Superoxide dismutase activity of ipsilateral testis was higher in torsion/detorsion and torsion/detorsion+erdosteine groups than control and torsion groups (p < 0.05). The biochemical parameters were not affected in controlateral testis in all groups. Torsion, torsion/detorsion and torsion/detorsion+erdosteine groups showed ipsilateral testicular damage in the histological examination, but the specimens from torsion/detorsion had a significantly greater histological injury than those from the other groups (p < 0.05). Control rats showed normal seminiferous tubule morphology. Rats in torsion group had slight-to-moderate disruption of the seminiferous epithelium. Rats in torsion/detorsion group displayed moderate-to severe disruption of the seminiferous epithelium. In all animals from torsion/detorsion+erdosteine group, the testicular tissues were affected with slight-to-moderate degenerative changes of the seminiferous epithelium. Administration of erdosteine resulted in a significantly reduced histological damage associated with torsion of the spermatic cord compared with torsion/detorsion. In all groups, the contralateral testes were histologically normal. In conclusion, the results clearly displayed that erdosteine treatment may have a protective role on testicular torsion/detorsion injury. PMID- 16311924 TI - Molecular cloning and expression of novel alternatively spliced cytochrome P450 2E1 mRNAs in humans. AB - Human cytochrome P450 2E1 (CYP2E1) is a phase I metabolizing enzyme. It is involved in the biotransformation of xenobiotics and endogenous substrates. Inter individual genetic polymorphisms of the CYP2E1 gene are associated with different cancer diseases as well as alcohol and nicotine dependence. We report here for the first time three novel alternative spliced mRNA transcripts which are more frequently present in lung carcinoma cell lines as in hepatocyte cell lines. They are unexpected detectable in blood leukocytes from healthy volunteers but not in normal and cancerous lung tissue. The full-length wildtype transcript of CYP2E1 is described to be concomitant to an alternatively spliced mRNA transcript. Stimulation with CYP2E1-inducing agents did not change the splicing transcript pattern. The three splicing variants should lead to truncated non-functional proteins. Thus the genetic diversity of CYP2E1 is additionally extended at the transcriptional level of gene expression. The physiological role of the splicing variants is not known, yet, but they seem to be related to the carcinogenic property of the cell lines. PMID- 16311925 TI - Radiation protection of DNA by ferulic acid under in vitro and in vivo conditions. AB - The effect of ferulic acid was studied on gamma-radiation-induced relaxation of plasmid pBR322 DNA and induction of DNA strand breaks in peripheral blood leukocytes and bone marrow cells of mice exposed to whole body gamma-radiation. Presence of 0.5 mM ferulic acid significantly inhibited the disappearance of supercoiled (ccc) plasmid pBR322 with a dose modifying factor (DMF) of 2.0. Intraperitoneal administration of different amounts (50, 75 and 100 mg/kg body weight) of ferulic acid 1 h prior to 4 Gy gamma-radiation exposure showed dose dependent decrease in the yield of DNA strands breaks in murine peripheral blood leukocytes and bone marrow cells as evidenced from comet assay. The dose dependent protection was more pronounced in bone marrow cells than in the blood leukocytes. It was observed that there was a time-dependent disappearance of radiation induced strand breaks in blood leukocytes (as evidenced from comet parameters) following whole body radiation exposure commensuration with DNA repair. Administration of 50 mg/kg body weight of ferulic acid after whole body irradiation of mice resulted disappearance of DNA strand breaks at a faster rate compared to irradiated controls, suggesting enhanced DNA repair in ferulic acid treated animals. PMID- 16311926 TI - Kinetic characteristics of folate binding to rat renal brush border membrane in chronic alcoholism. AB - The binding of folic acid to the plasma membrane is an important event for its reabsorption and conservation by renal epithelium. We studied [3H] folic acid binding to the renal cortical brush border membrane vesicles (BBMV) in rats after 12 weeks of chronic ethanol ingestion. Our results demonstrated that after chronic ethanol intake, the binding of folic acid to the membrane vesicles: (1) was decreased by a mechanism that decreased B(max) without affecting K(d), (2) was insensitive to Na+ ions in the medium (3) behaved differently to divalent cations in the medium in contrast to control group. However under such conditions there was no significant effect of ethanol on pH optimum of the process. Binding was reduced at pH less than 6 but there was no effect of ethanol on binding in pH range from 6 to 8. Increasing the osmolarity at pH 5.5 had no effect on the binding of folate to BBMV, thus confirming that the observed changes in B(max) values were due to site-specific binding in the two groups. Moreover, ethanol does not impart its effect on involvement of sulfhydryl group at the binding site of folate transport system. These findings highlight the possible mechanism of perturbed renal conservation of folate during chronic alcoholism. PMID- 16311927 TI - Splice isoform of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-4: expression and hypoxic regulation. AB - The 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFKFB) is responsible for maintaining the cellular levels of fructose-2,6-bisphosphate which is a key regulator of glycolysis. Here we have studied the expression of PFKFB-4 isozyme in the DB-1 melanoma cells. An additional isoform of PFKFB-4 mRNA with 148 bases insert in the amino-terminal region at high constitutive levels was identified in these cells. The expression of this splice isoform as well as main isoform of 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase was responsible to hypoxia and dimethyloxalylglycine, an inhibitor of HIF-1 alpha hydroxylase enzymes, suggesting that the hypoxia responsiveness of PFKFB-4 gene in these cells is regulated by HIF-1alpha protein. Hypoxic induction of PFKFB4 mRNA in the DB-1 melanoma cells correlates with the expression of PFKFB-3 and VEGF mRNA which are known as HIF-1 dependent genes. Thus, our results clearly demonstrated the existence of splice isoform of PFKFB-4 mRNA in the DB-1 melanoma cells and its overexpression under hypoxic conditions. PMID- 16311928 TI - Superoxide production and oxygen consumption in endothelium-intact and -denuded artery stimulated by angiotensin II. AB - Arteries stimulated by angiotensin II (AII) to contract do not display the expected augmentation of O2 consumption seen with other cardiovascular contractile agonists. We tested the hypothesis that superoxide (O2-) or other reactive oxidant species generated by AII played a role in the paradoxical O2 consumption response in porcine carotid artery, with or without an intact endothelium. Endothelium-denuded arteries were incubated with either 1 microM diphenylene iodonium (DPI), an inhibitor of NAD(P)H oxidase, 300 u/ml superoxide dismutase (SOD), a scavenger of O2-, or 20 U/ml catalase, an enzyme which promotes conversion of O2- (scavenged in the form of H2O2) to O2. DPI treatment resulted in the expected increase in O2 consumption upon contractile activation with AII challenge (1.05+/- 0.23 micromol/g/min; n = 6, p < .01), as did treatment with SOD (0.67+/- 0.20 micromol/g/min; n = 4, p < .05). Catalase incubation resulted in a burst of O2 generation upon AII challenge (1.30 +/- 0.21 micromol/g/min; n = 10, p < .001). In endothelium-intact arteries, O2 consumption was again not augmented with AII challenge; instead, a burst of O2 production was observed (0.66 +/- 0.22 micromol/g/min; n = 9, p < .05), which was not affected further by addition of catalase. Thus, the absence of apparent augmentation of O2 consumption during contractile activation of endothelium-denuded arteries was attributed to simultaneous NAD(P)H oxidase-dependent production of O2-, and attendant H2O2 and O2 generation which either and masked the detection of O2 consumed or suppressed mitochondrial uptake of O2, or both. An intact endothelium was required to manifest the burst of O2 generation with AII stimulation under normal conditions. PMID- 16311929 TI - Nicotine metabolism in healthy smokers and patients with cardiovascular diseases. AB - In this study, we measured the excretion rate of nicotine and its two major metabolites, cotinine and trans-3'-hydroxycotinine (THOC), in the urine of 25 healthy smokers and 15 smokers who underwent a coronary artery bypass surgery or coronary angioplasty. After 1 day of smoking cessation, urine samples were collected in the morning, before smoking two cigarettes, and then three times after smoking, approximately 4 h apart. The results show that (i) in healthy smokers, nicotine and its two major metabolites were present at high concentration in the first urine sample after smoking, (ii) in smokers with cardiovascular disease nicotine and cotinine were less excreted whereas THOC was more excreted, mainly in the second urine sample. We conclude that this shift in nicotine metabolism may contribute to smoking-induced cardiovascular disease. PMID- 16311932 TI - Spontaneous delivery following tension-free vaginal tape procedure. AB - There has been no report in the international literature concerning vaginal delivery following tension-free vaginal tape (TVT) procedure. Most gynecologists recommend cesarean section after TVT procedure. We present the case of a 37-year old (gravida 2, para 2) woman who had spontaneous delivery at 40 weeks' gestation after TVT procedure performed 10 months prior because of stress urinary incontinence. Five months after spontaneous delivery, the patient was shown to be continent, with no urinary leakage occurring following stress maneuver. Urodynamic evaluation showed normal urethral pressure profile and sufficient maximum urethral closure pressure. Introital ultrasound demonstrated the correct position of the Prolene tape. In cases of pregnancy following TVT procedure, a general recommendation of delivery by cesarean section may be questioned, since the function and correct suburethral position of the Prolene tape can also remain intact following vaginal delivery. PMID- 16311933 TI - Beyond heart rhythms: new directions for implantable devices. AB - Implantable cardiac devices have become firmly entrenched as important therapeutic tools for a variety of conditions. Pacemakers are the only available treatment for symptomatic bradycardia not due to reversible causes. Large randomized studies have demonstrated a small but statistically significant reduction in atrial fibrillation associated with pacing modes that maintain atrioventricular synchrony. In contrast, pacing mode appears to have a less dramatic effect in patients with atrioventricular block. Cardiac resynchronization with specialized left ventricular leads has been shown to reduce symptoms and improve survival in patients with symptomatic heart failure, systolic dysfunction, and widened QRS complexes. The implantable cardioverter defibrillator has become the standard therapy for protecting patients against sudden cardiac death. Two recent trials, Multicenter Automatic Defibrillator Trial II (MADIT II) and the Sudden Cardiac Death Heart Failure Trial (SCD-HEFT), demonstrated that the ICD is associated with a significant survival benefit for patients with reduced ejection fraction (< 0.30-0.35) particularly if heart failure symptoms are present. Finally the implantable loop recorder has become an important diagnostic tool for the patient with unexplained syncope. This brief overview summarizes the indications and follow-up of the wide array of implantable cardiac devices available to the clinical cardiologist. PMID- 16311934 TI - Post mortem analysis of a left atrial appendage occlusion device (PLAATO) in a patient with permanent atrial fibrillation. AB - Thromboemoblism is the most feared complication of atrial fibrillation. Percutaneous left atrial appendage occlusion is a new interventional procedure for reducing thromboembolic risk in patients with atrial fibrillation. The paper reports of a post mortem analysis of the device demonstrating that one year after implantation the atrial surface of the device is completely covered by neo endothelium and the device occludes the appendage completely. PMID- 16311935 TI - Incidence of pulmonary vein stenosis in patients submitted to atrial fibrillation ablation: a comparison of the Selective Segmental Ostial Ablation vs the Circumferential Pulmonary Veins Ablation. AB - INTRODUCTION: Pulmonary vein (PV) stenosis is an important complication of the AF ablation and could be underestimated if their assessment is not systematically done. Selective Segmental Ostial Ablation (SSOA) and Circunferential Pulmonary Veins Ablation (CPVA) have demonstrated efficacy in atrial fibrillation (AF) treatment. In this study the real incidence of PV stenosis in patients (pts) submitted to both SSOA and CPVA was compared. METHODS: Those pts with focal activity and normal left atrial size were submitted to SSOA, remaining pts were submitted to CPVA to treat refractory, symptomatic AF. Contrast enhanced magnetic resonance angiography (MRA) was routinely performed in all patients 4 months after the procedure. RESULTS: A series of 73 consecutive patients (mean age of 51 +/- 11 years; 75% male) were included. SSOA was performed in 32 patients, and the remaining 41 patients underwent to CPVA, obtaining similar efficacy rates (72% vs 76% arrythmia free probability at 12 months; log rank test p = NS). Six patients had a significant PV stenosis, all in SSOA group none in CPVA group (18.8% vs 0%; p = 0.005). All patients were asymptomatic and the stenosis was detected in routine MRA. No predictors of stenosis has been identified analysing patient procedure characteristics. CONCLUSION: PV stenosis is a potential complication of SSOA not seen in CPVA. The study confirms than MRA is useful for identifying patients with asymptomatic PV stenosis. PMID- 16311936 TI - Comparison of modern steroid-eluting epicardial and thin transvenous pacemaker leads in pediatric and congenital heart disease patients. AB - OBJECTIVE: Optimal pacemaker lead choice in pediatric patients eligible for either epicardial or transvenous leads remains unclear. We compared performances of modern thin transvenous (TTV) and steroid-eluting epicardial (SEE) leads in patients followed at one pediatric center. METHODS: Retrospective review of patients with qualifying leads implanted from August 1997 to March 2004. Threshold energy (TE) at implant and follow-up, sensing thresholds, lead complications, and repeat pacing-related procedures were analyzed. Lead performances were compared using t-tests, Wilcoxon rank-sum tests and Cox regression. Survival curves were plotted using Kaplan-Meier analysis. RESULTS: A total of 370 implant procedures, 521 leads, and 1549 visits were evaluated. In all, 256 leads were SEE (49%, 184 implants) and 265 were TTV (51%, 186 implants). Median follow-up was 29 months (range 1-80 months). Patients with SEE systems were younger at implant (6 vs. 17 yrs, p < 0.001), and more had congenital heart defects (82% vs. 57%, p < 0.001). At follow-up, ventricular TEs were higher for SEE leads at implant (p < 0.001), 1 month (p < 0.001), and up to 4 years (p = 0.019). When compared across all follow-up durations combined, TTV TEs were significantly lower than SEE TEs for both atrial and ventricular leads (p < 0.001). A total of 70 repeat procedures were performed in 60 patients during the study period, which comprised 18% of SEE and 14% of TTV system patients (p = NS). In all, 18 TTV and 19 SEE leads failed (p = NS). Estimated freedom from lead failure at 1, 3, and 5 years was 97%, 88%, 85% for TTV leads and 96%, 92%, and 58% for SEE leads (log rank P = NS). CONCLUSIONS: Both SEE and TTV leads showed good mid-term performance and survival in our cohort. Higher TEs seen for SEE leads, especially ventricular and unipolar leads, may result in higher current drain and thus more generator replacements than TTV systems. Lead failure rates were comparable across lead types. TTV leads offer a promising alternative to SEE systems in terms of performance for young patients without intracardiac shunting who do not require open-chest surgery for another indication. PMID- 16311937 TI - Fractally coated myocardial pacemaker leads in children. AB - We report our single centre experience with a new fractally coated myocardial unipolar lead (ELC35UP; Biotronik) in 96 pediatric patients (59% male, 41% female). Congenital heart disease (CHD) was associated in 89%. The age at implantation ranged between 2 days and 19 years, median for children with CHD 7.8 years, without CHD 4.7 years. Twenty percent of the children were younger than one year at implantation. Mean follow-up was 30 months (1-57 months). We compared our findings with a steroid eluting epicardial lead (CapSure EPI 4968; Medtronic) in 46 children with comparable age and sex-distribution. We found a lead survival of 87% after 57 months in the ELC35UP group (steroid lead: 87% after 129 months). Pacing energy thresholds were equal after 12 months (median 3.0 microjoules), but the sensing characteristics of the fractally coated lead was significantly superior to the steroid eluting lead with median R waves of 7.0 mV (steroid lead: 3.5 mV) after 12 months. Children with myocardial scar tissue requiring pacemaker therapy after surgery of CHD showed no differences in sensing and pacing thresholds in comparison to children with congenital rhythm disorders. The fractally coated screw-in lead offers technical advantages concerning the subxiphoidal implantation procedure. CONCLUSION: Fractally coated ventricular screw-in leads represent a feasible alternative to the common steroid eluting leads- especially in children requiring pacemaker therapy after surgery for CHD. PMID- 16311938 TI - Coronary sinus flow reversal in congestive heart failure detected during biventricular pacing. AB - This report describes a phenomenon observed during implantation of a left ventricular lead through the coronary sinus. During coronary sinus venography there was reflux of dye from the coronary sinus ostium towards the distal part of the sinus against the anticipated direction of flow. This was correlated to elevation of the right atrial pressure above the pressure within the coronary sinus. A review of data is provided and suggests several deleterious effects of this phenomenon on left ventricular function. PMID- 16311939 TI - Echocardiography induced tachycardia in a patient with a minute ventilation rate responsive pacemaker. AB - We report a case of a 57 year-old man with a minute ventilation rate-response pacemaker and an inappropriate increase in his heart rate during a transthoracic echocardiography study. The increase in heart rate was due to interference between echocardiography and minute ventilation sensor. This was confirmed by a repeat echocardiogram, which reproduced the same effect and turning off the minute ventilation sensor stopped the tachycardia. We suggest that deactivating the minute ventilation sensor would avoid this interaction and electrophysiologists should be aware of this unusual electromagnetic interference phenomenon. PMID- 16311940 TI - Septic shock induced from an implantable cardioverter-defibrillator lead associated Candida albicans vegetation. AB - In recent years, the use of implantable cardioverter-defibrillators has made a significant impact in the treatment of cardiac disease. With the advent of this therapy, infection has become a growing concern, with a reported incidence of 1 11%. Although device-related infections are most frequently caused by bacterial pathogens, fungal etiologies are uncommon and carry an exceedingly high mortality rate. To date, there is only one published report of an implantable cardioverter defibrillator lead-associated infection secondary to Candida albicans. This case report presents the second such case. A review of the literature regarding fungal endocarditis as well as diagnosis and therapy of this rare and dangerous disease is discussed. PMID- 16311941 TI - Cardiac glycosides: William Withering (1741-1799). PMID- 16311944 TI - Fetal origins of adult disease: a paediatric perspective. AB - Prepubertal children born SGA or VLBW premature exhibit marked insulin resistance. There are similarities between SGA and VLBW children in that both are exposed to sub-optimal environments that encompass undernutrition and/or malnutrition during the equivalent of the last trimester of pregnancy. Although both SGA and VLBW groups fail to reach genetic height potential and are recognized causes of short stature in childhood, there are differences between the groups with respect to the growth hormone and IGF-I axis.SGA children have elevated IGFI levels, possibly due to either hyperinsulinism or partial IGF-I resistance, whereas VLBW children have low IGF-I and IGFBP-3 levels suggestive of GH resistance. Thus the nature and timing of the early insult may lead to discordant changes to the metabolic and endocrine axes.IVF children are taller with increased IGF I, IGF II and IGFBP3 expression. These changes could be due to alterations in the environment of the periconceptual embryo resulting in changes in imprinting of genes involved in growth and development. The phenotypic, endocrine and metabolic consequences of alterations in the periconceptual, fetal and early neonatal periods is an area of intense investigation. Future research in this field is likely to focus on the mechanisms through which environmental changes lead to these programmed effects. PMID- 16311943 TI - Pediatric endocrine disorders of energy balance. PMID- 16311945 TI - New issues in the diagnosis and management of Turner syndrome. AB - This review has tried to update our view of TS, highlighting the less severe phenotype we are seeing today, aiming to motivate clinicians to scrutinize normal looking short girls more closely, and to provide more relevant information for those counseling prospective parents on the implications of a TS karyotype during prenatal screening. New approaches to cardiac evaluation, including imaging with MR and ECG analysis-were suggested to strengthen our ability to detect and prevent potentially life-threatening cardiac complications. The new emphasis on reproductive potential and concerns about the adequacy and safety of current HRT regimens certainly require further studies and adjustment of treatment strategies in light of new priorities and safety concerns. In the same vein, prospective studies are required to assess the outcome and safety of assisted pregnancy in TS, which, despite the warning ofa potential catastrophic increase in maternal morbidity is going to become a much more common occurrence in the near future. PMID- 16311946 TI - Bone mineral accrual and low bone mass: a pediatric perspective. PMID- 16311947 TI - New Insights into biological roles of relaxin and relaxin-related peptides. PMID- 16311948 TI - Hyperglycemic comas in children: new insights into pathophysiology and management. PMID- 16311949 TI - [Uveal melanoma: the challenge to save more lives]. PMID- 16311950 TI - [NSAIDs in ocular inflammation treatments]. PMID- 16311951 TI - [Blindness and other eye diseases in Galdos' novels]. PMID- 16311952 TI - [Chronic blepharitis and Demodex]. AB - PURPOSE: Demodex is a mite commonly found in eyelash hair follicles and sebaceous glands of healthy people. Due to the fact this mite has also been reported in many chronic cases of blepharitis, we aimed to investigate the incidence of infestation with this mite in healthy people and in patients with chronic blepharitis, in addition to evaluating the response to the different treatments used for blepharitis. METHODS: 105 subjects without blepharitis were selected as a control group and 20 subjects diagnosed as having chronic blepharitis were selected as patients. Epilated eyelashes were observed under microscope in both groups of patients. Those with overpopulation of mites were treated with ether lid and eyelash scrubs and 2% mercury oxide ointment. RESULTS: The incidence of Demodex infestation in the control group was 0.08 mites per eyelash, whereas in the patients with chronic blepharitis the incidence was 0.69 mites per eyelash; this difference was statistically significant (p=0.006). All the mites found were Demodex folliculorum except in one patient where the mite was identified as Demodex brevis. After 3-8 weeks of specific treatment the number of mites per eyelash decreased dramatically (0.03, with p=0.001). Two patients were intolerant of the therapy. CONCLUSIONS: In this study the incidence of Demodex in patients with blepharitis was very high, when compared with normal subjects, showing a clear association between blepharitis and Demodex infestation. Treatment with mercury oxide ointment was satisfactory in controlling the infection, despite difficulty in its application and occasional toxicity. PMID- 16311953 TI - [Comparative study between the non-contact pneumotonometer Canon TX10 and the Goldmann tonometer]. AB - PURPOSE: To study and contrast the measurement of intraocular pressure (IOP) with an air non-contact tonometer (Canon TX-10) with that obtained with the Goldmann tonometer. METHODS: Intraocular pressure was measured in 649 subjects with both the pneumotonometer and the Goldmann tonometer. Statistically significant differences between the results obtained by both tonometers and the influence of variables including age, corneal thickness, and corneal curvature were studied. RESULTS: The mean age of the study population was 48.5 years, standard deviation (SD) 7.6 years, with a range of 22-68 years. The mean IOP was 15.4, SD 2.7 mm Hg, (range 8-25) with the Goldmann tonometer and 15.6, SD 2.9 mm Hg (range 8-29) with the pneumotonometer. There was a statistically significant difference between the measurements using the air pneumotonometer and the Goldmann tonometer (p<0.001). The mean of the differences between both tonometers was 0.24, SD 2 mm Hg. Seventy five percent of measurements were in the range of +/-1, 85% in +/-2, and 95% in the Bland and Altman's interval of 4.42, -3.94. These differences were more significant for higher values of IOP, in thicker corneas and in corneas with greater curvature. A correlation between IOP and age was not found (p=0.247). CONCLUSIONS: The pneumotonometer is a useful and reliable instrument for measuring the IOP providing there is additional checking in cases with borderline values of IOP, or excessive corneal thickness and/or curvature. PMID- 16311954 TI - [Contrast sensitivity to intraocular lens TECNIS Z-9000]. AB - PURPOSE: To evaluate the increase in contrast sensitivity to intraocular lens TECNIS Z-9000 (Pfizer) compared with other intraocular lenses. METHOD: We implanted a TECNIS Z-9000 lens in 18 patients who had another type of intraocular lens implanted in their other eye. We then tested contrast sensitivity under both mesopic and photopic conditions using the VCTS (Vision Contrast Test System) provided by Vistech Consultants Inc. RESULTS: There was an improved contrast sensitivity at low and medium frequencies, following TECNIS-Z 9000 lens implantation, however this improvement did not reach statistical significance. There were significant differences at high frequencies, which are directly related to the quality of visual function. CONCLUSIONS: TECNIS Z-9000 lens implantation (with a modified anterior surface) achieves a significant improvement in contrast sensitivity, meaning in practice an increase in the quality of visual function. PMID- 16311955 TI - [Late endophthalmitis following Ahmed valve]. AB - CASE REPORT: A 71-year-old woman with a history of aphakic glaucoma underwent implantation of an Ahmed valve and scleral grafting in her right eye. Postoperative visual acuity was 0.5 and intraocular pressure was 12 mmHg during treatment with brimonidine tartrate (0.2%). Nine months after implantation she suffered a conjunctival infection which was treated with hygienic measures and topical antibiotic therapy. Four days later, she developed an endophthalmitis which was treated with topical, intravitreous and intravenous vancomycin and ceftazidime. The Ahmed drainage implant was replaced at 72 hours. Laboratory culture yielded Haemophilus influenzae. Four days later, the eye was enucleated. DISCUSSION: Endophthalmitis is an uncommon complication of glaucoma drainage implant surgery. Exposure of the drainage tube represents the greatest risk factor for this condition. Removal of the implant in the first 24 hours is recommended if a good visual prognosis is to be achieved. PMID- 16311956 TI - [Severe bilateral ocular affection caused by vitamin A deficiency]. AB - CASE REPORT: We report a patient with alcohol-induced chronic pancreatitis, with associated caloric-protein malnutrition and lipid soluble vitamin deficiencies, along with peripheral ulcerative keratitis in the right eye and necrotizing stromal ulceration with hyphema in the left eye. DISCUSSION: Although uncommon in developed countries, Vitamin A deficiency must be considered when there is a bilateral corneal affection in alcoholic patients or in patients with gastrointestinal, pancreatic or biliary pathology. PMID- 16311957 TI - [Malignant glaucoma following combined Ahmed valve implant and phacoemulsification surgery for chronic angle-closure glaucoma]. AB - CASE REPORT: Simultaneous Ahmed valve implant, combined with phacoemulsification cataract surgery, is a useful therapeutic option for patients with chronic angle closure glaucoma, when conventional filtering surgery fails. This combined approach permits both control of intraocular pressure and early recovery of visual function. We report the results in five patients successfully treated with this combined procedure, two of whom developed early postoperative malignant glaucoma. DISCUSSION: Predisposing anatomic features in patients with chronic angle-closure glaucoma, associated with sudden anterior chamber decompression and increased postoperative inflammation, may facilitate the development of malignant glaucoma following combined glaucoma implant and phacoemulsification surgery. PMID- 16311958 TI - [Corneal intra-epithelial neoplasia (CIN). Treatment by combined excision and antimetabolite therapy]. AB - CASE REPORT: We present two clinical cases of corneal intra-epithelial neoplasia defined histopathologically. DISCUSSION: We sought to establish the importance of treatment with Mitomycin-C in two cases of corneal intra-epithelial neoplasia. The differential diagnosis was established based on the evolutionary character of the process, as well as specific histopathological aspects of these entities. In both cases, treatment with Mitomycin-C before, during and after surgery contributed to the decrease in the size of the lesion and to their cure without relapse. PMID- 16311959 TI - [Asymptomatic vasoproliferative retinal tumor]. AB - PURPOSE/METHOD: A case of vasoproliferative retinal tumor is described. The main differential diagnoses must be excluded and the therapeutic possibilities defined. RESULTS/CONCLUSIONS: Vasoproliferative retinal tumors are infrequent benign retinal lesions of gliovascular composition. The differential diagnoses needing consideration are a retinal hemangioma associated with von Hippel-Lindau disease, an amelanotic choroidal melanoma and an ocular metastasis from a tumor elsewhere. Treatment must be performed in those cases where the sight is threatened. PMID- 16311960 TI - [SCA-7. Cone-rod dystrophy in the context of an hereditary ataxia]. AB - CLINICAL CASE: A 21-year-old male presented with bilateral loss of visual acuity within the last year, and cerebellar ataxia since childhood. Two members of his family had a similar disorder. Examination showed bilateral central scotomas, as well as an electroretinogram pattern and optic coherence tomography images consistent with cone-rod dystrophy. Molecular analysis by PCR amplification and genotyping of the SCA7 gene established the diagnosis of SCA-7. DISCUSSION: SCA-7 is a polyglutamine expansion disorder and the only spinocerebellar ataxia that shows a cone-rod dystrophy phenotype, which probably results from interference with the action of specific cone-rod genes. PMID- 16311961 TI - [Microscopic study of the living eye]. PMID- 16311962 TI - [Convex mirrors in Flemish painting (I)]. PMID- 16311968 TI - Low weight in congenital heart surgery: is it the right way? AB - OBJECTIVE: There is ample evidence that premature and low birth weight children have a poor outcome after congenital heart surgery. The aim of the present study was to characterize the perioperative factors which significantly influence the outcome of these babies following cardiac surgery, and to clarify whether the RACHS-1 and the Aristotle score are compatible for this complex kind of heart surgery. METHODS: During the past 10 years, 108 children with a body weight of less than 3000 g were operated, including 43 premature babies. Mean weight at operation was 2.5 +/- 0.5 kg, mean age was 36.8 +/- 55 days. Fifty percent of the treatments were categorized into risk groups 4 and 6 and 54 % belonged to the complexity levels 3 and 4. Eighty-one operations (75 %) were performed using extracorporeal circulation. RESULTS: The 30 day mortality rate was 18 %, and the overall mortality rate was 30 %. The correlation between mortality rates and risk groups was significant ( p < 0.001). Other significant factors were preoperative acidosis ( p = 0.026), preoperative catecholamine support ( p < 0.001), prolonged ICU stay (> 7 days) after operation ( p < 0.001), and postoperative infection ( p = 0.019). In addition, X-clamp time ( p = 0.029) and palliative procedures ( p < 0.001) were significant factors for poor outcome. CONCLUSION: The results demonstrate that the mortality for correction of congenital heart defects in children weighing less than 3000 g depends on several factors. The risk groups of the RACHS-1 study and the complexity levels of the Aristotle score are useful tools to assess preoperative risk. PMID- 16311969 TI - Selective cerebral perfusion via right axillary artery direct cannulation for aortic arch surgery. AB - BACKGROUND: The risk of neurological complications is still a life-threatening event for patients undergoing proximal aortic arch or total aortic arch surgery. To prevent these complications, axillary artery cannulation and antegrade selective cerebral perfusion were utilized. We compared the effects of using hypothermic circulatory arrest (HCA) alone or with selective cerebral perfusion (SCP/AX) via right side axillary artery direct cannulation. METHODS: 120 patients, mean age 61 +/- 12 years (range 26 - 80), underwent proximal aortic or total aortic arch replacement between 1999 and 2004; 46 were female. We retrospectively compared the results of the two patient groups comparable for preoperative risk factors: 71 pts were operated using HCA beginning in 1999 and 49 pts using HCA/SCP via axillary artery direct cannulation since 2002. The indication for surgery was an aortic aneurysm in 80 (67 %) patients and aortic dissection in 36 (30 %) patients. The groups were well matched with regard to median age (60 vs. 62 yrs), urgency (emergent/urgent 36 vs. 44 %; elective 64 vs. 65 %), and several other known risk factors ( p = ns). RESULTS: Overall in hospital mortality was 13 %: 10 % with HCA vs. 6 % with SCP/AX. Permanent neurological dysfunction occurred in 10 % with HCA vs. 6 % with SCP/AX. Transient neurological dysfunction (TND) in patients surviving without stroke was lower with SCP/AX (10 %) than with HCA (17 %) ( p = ns). Mean duration of HCA was 28 +/ 12 min when isolated HCA was used, and significantly shorter with 21 +/- 6 min when the combination of SCP/AX ( p = 0.03) was used. Mean duration of CPB was 202 +/- 55 min with HCA vs. 192 +/- 50 min with SCP/AX ( p = ns). Comparison of the groups who had comparable preoperative risk factors showed a trend towards lower in-hospital mortality, stroke and TND rates, a significant reduction in cardiac ( p = 0.034), infectious ( p = 0.025) and bleeding complications ( p = 0.04) in SCP/AX compared with HCA, as well as a significantly shorter duration of hospitalization ( p = 0.046) and shorter ICU stay ( p = ns). CONCLUSION: Our results suggest that HCA/SCP is superior to HCA alone for preventing cerebral injury during operations on the aortic arch. By reducing embolic risk, as well as the duration of HCA, SCP with axillary artery direct cannulation may be the optimal technique for averting cerebral events, reducing complications, and shortening hospital stays following aortic arch repair. PMID- 16311970 TI - Aspirin and clopidogrel taken until 2 days prior to coronary artery bypass graft surgery is associated with increased postoperative drainage loss. AB - OBJECTIVE: Platelet aggregation inhibitors, such as aspirin and clopidogrel, are associated with increased bleeding in patients undergoing cardiac surgery with cardiopulmonary bypass. We investigated the impact of time between the last intake of aspirin and clopidogrel before CABG surgery and drainage loss, transfusion requirements and rate of reoperation. PATIENTS AND METHODS: The records of patients who had coronary artery bypass graft surgery (CABG) were reviewed for intake of aspirin and clopidogrel within 7 days prior to surgery. Drainage loss, transfusion requirements and rate of reoperation for bleeding within 5 days after the operation, were recorded. RESULTS: Out of 261 analysed patients, 225 patients (86.2 %) had no anti-platelet medication and 36 patients (13.8 %) were on aspirin and clopidogrel. Aspirin and clopidogrel, taken all until 2 days prior to operation, were associated with a significantly higher postoperative blood loss (1840 mL [1230 - 3710] vs. 280 mL [185 - 765], p = 0.005 for one day and 850 mL [345 - 1725] vs. 277 mL [165 - 778], p = 0.026, for 2 days prior to surgery). The trend showed that patients in the study group received more platelet concentrates (PC: 5.3 % vs. 13.9 %, p = 0.067). The rate of reoperation for bleeding was not different ( p = 0.25). CONCLUSION: Aspirin and clopidogrel up to 2 days prior to CABG were associated with a significantly higher postoperative drainage loss. PMID- 16311971 TI - Contrast echocardiography: a valid and sensitive imaging method in the investigation of transmyocardial laser-revascularization in an acute ischemia model. AB - In an acute porcine myocardial ischemia model we examined the effect of transmyocardial laser-revascularization (TMLR) on the regional micro perfusion and oxygen supply. In clinical practice, contrast echocardiography is a reliable tool for the measurement of changes in regional blood flow in the ischemic myocardium. We compared contrast echocardiography with the laser Doppler measurement of micro perfusion and with the quantification of regional tissue oxygen tension using a Clark electrode. 22 pigs were randomised in the interventional group and 12 in the control group. Measurements with all three methods were performed before and after occlusion of the first diagonal branch of the left anterior descending coronary artery and, in the interventional group, after TMLR. We investigated not only the effects in the dependent ischemic myocardium, but also in two other myocardial areas, not involved in the coronary occlusion. There was a significant effect of TMLR on regional micro perfusion and local oxygen tension in the dependent ischemic myocardial area. Contrast echocardiography is a successful experimental tool to measure changes in regional myocardial perfusion which cannot be perceived using clinical imaging methods. PMID- 16311972 TI - Protecting the endothelial integrity of internal thoracic arteries. AB - BACKGROUND: Previous functional studies on human internal thoracic arteries, comparing the effect of the traditional harvesting method (occlusion with a clip) with a method leaving the artery perfused, revealed considerably impaired endothelial function associated with enhanced contractility after clipping. We have now investigated whether these observations could be correlated (1) with plasma markers of endothelial dysfunction, and (2) with structural changes in the endothelial layer. METHODS: 32 patients were randomly distributed into groups of clipped and perfused arteries. Arterial blood samples were obtained from both the artery and extracorporeal circulation to determine sP-selectin, sE-selectin, sL selectin, and thrombomodulin using enzyme-linked immunosorbent assay. Arteries from three patients were examined by scanning electron microscopy. RESULTS: Concentrations of sP-selectin and thrombomodulin were significantly higher in plasma from clipped arteries compared to perfused arteries, whereas sE-selectin and sL-selectin concentrations were similar within the groups. Scanning electron microscopy revealed significant structural changes and loss of endothelial cells in clipped arteries. CONCLUSION: Biochemical and structural results support our findings that leaving the internal thoracic artery perfused preserves endothelial function in the arterial graft. PMID- 16311973 TI - Characteristic differences between patients who have undergone surgical treatment for lung metastasis or hepatic metastasis from colorectal cancer. AB - OBJECTIVE: The characteristic differences between patients with lung or liver metastases from colorectal carcinoma (CRC) have not yet been clarified. A small group of these patients demonstrate a better prognosis, and the selection criteria for resection of liver and/or lung metastasis are not well defined. It is important to compare and analyze the most common metastatic sites, which include liver metastases and lung metastases. The objective of this study was to compare the characteristics of the two groups in order to identify patients who benefitted from surgical resection of CRC. METHODS: We retrospectively reviewed the medical charts of 80 patients who had undergone resection for liver or lung metastasis from CRC in Fukuoka University Hospital between June 1991 and December 2004. These patients were grouped according to surgical therapy received for the metastases, and separated into two groups, as follows: LUM, lung metastases resection; LIM, liver metastases resection. We evaluated these groups for a set of several factors. RESULTS: The characteristic factors between the two groups (LUM vs. LIM) demonstrated significant differences according to histological differentiation, venous invasion, and lymphatic permeation. There was a statistical difference in the disease-free interval (DFI) between the two groups (947.06 +/- 840.39 days in LUM vs. 246.03 +/- 229.26 days in LIM). Although serum CEA levels at resection of metastasis showed significant differences between the groups (LUM, 13.25 +/- 31.55 ng/ml; LIM, 55.21 +/- 99.52 ng/ml), the primary serum CEA levels were not significantly different. Overall survival rates at 5 years were 37.0 % for LUM and 42.8 % for LIM. There was no significant difference in the survival rate of the LUM vs. the LIM group after resection of metastasis. The Cox proportional hazards regression model was used to determine serum CEA status at the time of the metastases and showed a significant difference indicating poor prognosis for patients with LUM, but the results were not significant for LIM cases. CONCLUSIONS: Candidates for surgical treatment for lung or liver metastases from CRC may be an acceptable for the same valuable approach, even if characteristic differences were observed in each group. PMID- 16311974 TI - Gastropericardial fistula and Candida kruzei pericarditis following laparoscopic Nissen fundoplication (gastropericardial fistula). AB - We report a case of gastropericardial fistula and Candida kruzei pericarditis one year after laparoscopic Nissen fundoplication. Chest X-ray and CT revealed pneumopericardium. Barium swallow, UGI, endoscopy, and bronchoscopy were negative. Pericardial exploration was performed through a sternotomy. Intraoperative fistulogram revealed a tract into the stomach. A midline abdominal incision allowed identification of the slipped Nissen, resection of the fistula tract, and subsequent re-do fundoplication. The patient was treated with amphotericin and remains symptom-free 10 months after her operation. We recommend both sternotomy and midline abdominal incisions to explore and access the pericardium, stomach, esophagus, and diaphragm. PMID- 16311975 TI - Prediction of morbidity after lung resection in patients with lung cancer using fuzzy logic. AB - BACKGROUND: Preoperative evaluation of patients with potentially resectable non small cell lung cancer aims to estimate the risk of planned surgery. Evidence of several factors that identify patients at risk for complications from thoracotomy is controversial. The aim of this study was to introduce and implement in medical practice a fuzzy system used in risk assessment of pulmonary resection for lung cancer. METHODS: Ninety-one consecutive patients who underwent pulmonary resection for lung cancer were investigated. The overall complication rate was 39.6 % (a total of 63 complications were seen in 36 patients). A fuzzy logic model was created with 9 input (presence of chest pain, weight loss, clinical T stage of the tumor, FEV 1, serum protein, preoperative arterial partial oxygen pressure and cigarette smoking, erythrocyte sedimentation rate and peripheral blood leukocyte count) and two output classes (high-risk and low-risk groups). The fuzzy classifier's performance was tested. RESULTS: The model was able to predict correctly the occurrence of complications in 22 out of 29 patients in the high-risk group with a sensitivity of 76 %, while 9 out of the 52 patients from the low-risk group developed complications (17 %). CONCLUSION: The fuzzy classification system provides an accurate tool to predict complications of resections in patients with non-small cell lung cancer. PMID- 16311976 TI - Extended resection of T4 lung cancer with invasion of the aorta: is it justified? AB - BACKGROUND: We report our 10-year experience of performing surgical resection of T4 lung cancer invading the thoracic aorta. PATIENTS AND METHODS: From 1994 to 2004, sixteen patients with T4 primary lung cancer with local invasion of the thoracic aorta underwent tumor resection. Surgical resection included 8 pneumonectomies and 8 lobectomies. The histologic type was squamous cell carcinoma in 7 patients, adenocarcinoma in 7, large cell carcinoma in 1, and small cell carcinoma in 1. Complete resection of the tumor with mediastinal lymph node dissection was achieved in 8 patients (50 %), while the resection was incomplete in the other 8 cases. RESULTS: The overall cumulative survival of the 16 patients at 3 and 5 years was 34.7 % and 17.4 %, respectively. The survival of the patients in the complete resection group was found to be 36.5 % at 5 years, with 2 patients surviving more than 5 years without a recurrence, which was significantly better than that of the incomplete resection group ( p = 0.005). CONCLUSIONS: Extended aortic resection with primary lung cancer is complex and possibly high risk, but can achieve long-term survival in selected patients. Surgical resection should be considered as a treatment option for T4 lung cancer for this T4 subcategory. PMID- 16311977 TI - Delayed esophageal necrosis and perforation secondary to thoracic aortic rupture: a case report and review of the literature. AB - Delayed esophageal necrosis secondary to aortic rupture is extremely rare but potentially fatal. Although diagnostic techniques have improved, survival remains rare. The clinical and imaging features have not been characterized, as no large series have been reported since the advent of computed tomography (CT) and endoscopy. Moreover, as delayed esophageal necrosis secondary to aortic rupture is rarely anticipated, diagnosis is usually delayed. We recently encountered a case of this complication and present this along with a literature review, in order to facilitate early recognition and treatment. In many cases, hemodynamics remain relatively stable despite aortic rupture and the long interval between onset and operation. Dysphagia is therefore an important symptom that may indicate compression of the esophagus and subsequent esophageal necrosis. Preoperative total obstruction of the esophagus on barium swallow, endoscopy, or CT is also an important feature suggesting ischemic change of the esophagus. Endoscopy often reveals an annular ulcer suggestive of an ischemic process leading to necrosis. Death generally occurs from severe mediastinitis occurring after graft repair of the aorta. We recommend performing postoperative endoscopy after aortic surgery when preoperative obstruction of esophagus is found. If endoscopy reveals annular necrosis, surgical treatment involving esophagectomy and omental translocation should be performed expediently before perforation occurs. PMID- 16311978 TI - Neoadjuvant chemotherapy and pneumonectomy in a lung cancer patient on hemodialysis. AB - Major pulmonary resections are rarely performed in non-small cell lung cancer patients on hemodialysis. To date only two cases of pneumonectomy performed in such patients are reported in the literature. Moreover, chemotherapy, as a treatment for advanced non-small cell lung cancer, is not routinely administered to patients with end-stage renal disease requiring hemodialysis. We present the case of a stage IIIB non-small cell lung cancer patient on hemodialysis who successfully underwent neoadjuvant chemotherapy followed by pneumonectomy. To our knowledge, this is the first case of non-small cell lung cancer patient on hemodialysis reported in the literature who successfully underwent this type of combined therapy. PMID- 16311979 TI - Extended aortic bypass for atherosclerotic occlusive disease of high abdominal aorta. AB - We describe a case of acute renal failure caused by extension of an atherosclerotic occlusive process to the suprarenal aorta. A 68-year-old man underwent bypass grafting from the ascending aorta to the femoral arteries using straight and Y-grafts. Additional saphenous vein grafts were placed from the intra-abdominal portion of the graft to the left and right renal arteries and the splenic artery. As a consequence, relief of limb ischemia was achieved and dialysis could be discontinued. PMID- 16311980 TI - Primary chylopericardium. AB - Primary chylopericardium is a rare entity. Here we describe a 36-year-old, asymptomatic male in whom pericardial effusion was detected by chest X-ray and echocardiography on routine health control. After pericardiocentesis that revealed the chylous nature of the fluid, partial pericardiectomy without duct ligation was carried out. In the follow-up period, there was no evidence of pericardial fluid on chest X-ray and echocardiography, at three months after the procedure. PMID- 16311981 TI - Successful use of recombinant factor VIIa in a patient with intractable bleeding during extracorporeal membrane oxygenation. AB - Bleeding is still the most common complication during extracorporeal membrane oxygenation (ECMO) for temporary cardio-circulatory support. We present a case of a young man suffering from intractable hemorrhage during ECMO support, who was pre-treated with glycoprotein IIb/IIIa receptor antagonist Tirofiban due to a suspicion of myocardial ischemia. After failure of conventional hemostatic means, hemostasis was achieved by the donation of recombinant Factor VIIa (rFVIIa). Aspects of bleeding control during extracorporeal circulatory support, the use of Tirofiban and rFVIIa are discussed. PMID- 16311982 TI - Cardiac surgery in Germany during 2004: a report on behalf of the German Society for Thoracic and Cardiovascular Surgery. AB - All cardiac surgical procedures performed in 79 German cardiac surgical units in the year 2004 are presented in this report, which is based on a voluntary registry organized by the German Society for Thoracic and Cardiovascular Surgery. A total of 100 830 cardiac surgical procedures (ICD and pacemaker procedures excluded) were reported to the registry for the year 2004, an increase by 1.1 % compared to the year 2003. More than 7.8 % of the patients were older than 80 years. Hospital mortality in 58 144 isolated CABG procedures (7.1 % off pump) was 2.8 %, and 4.5 % in 18 617 isolated valve procedures. This registry is an important tool of the German Society for Thoracic and Cardiovascular Surgery to allow a continuous and voluntary monitoring of quality and to illustrate the development of cardiac surgery in Germany. PMID- 16311985 TI - Tamoxifen-treated breast carcinoma patients and the risk of acute myocardial infarction and newly diagnosed angina. PMID- 16311986 TI - Successes and failures of the teachable moment: smoking cessation in cancer patients. AB - BACKGROUND: Successful cancer treatment can be significantly compromised by continued tobacco use. Because motivation and interest in smoking cessation increase after cancer diagnosis, a window of opportunity exists during which healthcare providers can intervene and assist in the quitting process. METHODS: The authors conducted a comprehensive literature review to discuss 1) the benefits of smoking cessation in cancer patients, 2) current knowledge regarding smoking cessation interventions targeted to cancer patients, and 3) treatment models and state-of the-art guidelines for intervention with cancer patients who smoke. The authors present clinical cases to illustrate the challenging nature of smoking cessation treatment for cancer patients. RESULTS: Continued smoking after cancer diagnosis has substantial adverse effects on treatment effectiveness, overall survival, risk of second primary malignancies, and quality of life. Although some encouraging results have been demonstrated with smoking cessation interventions targeted to cancer patients, few empirical studies of such interventions have been conducted. A range of intervention components and state of-the-art cessation guidelines are available that can be readily applied to cancer patients. Case illustrations highlight the crucial role of healthcare providers in promoting smoking cessation, the harmful impact of nicotine addiction manifested in delayed and failed reconstructive procedures, and unique problems encountered in treating patients who have particular difficulty quitting. CONCLUSIONS: Despite the importance of stopping smoking for all cancer patients, the diagnosis of cancer is underused as a teachable moment for smoking cessation. More research is needed to empirically test cessation interventions for cancer patients, and attention must be given to complex and unique issues when tailoring cessation treatment to these individuals. PMID- 16311987 TI - Treatment of local breast carcinoma in Florida: the role of the distance to radiation therapy facilities. AB - BACKGROUND: Breast-conserving surgery combined with radiation (BCSR) is the recommended alternative treatment to mastectomy for local breast carcinoma. However, limited access to healthcare may result in more extensive surgical treatment. The effect of distance to radiation therapy facilities on the likelihood of receiving BCSR was examined in Florida. METHODS: Local breast carcinomas reported to Florida's statewide registry between July, 1997, and December, 2000 were linked to the Agency of Healthcare Administration inpatient and outpatient databases to supplement the registry's treatment data, resulting in 18,903 local breast carcinoma cases treated with BCSR or mastectomy. The odds of receiving BCSR were modeled as a function of distance to the closest radiation therapy facility, adjusting for health insurance, age, race/ethnicity, and marital status. RESULTS: Distance to the closest radiation therapy facility was negatively associated with BCSR, with the odds ratio (OR) decreasing by 3% per 5 mile increase in distance. Compared with the uninsured, privately insured women were 49% more likely to receive BCSR (OR of 1.49; 95% confidence interval [95% CI], 1.20-1.86) and Medicare patients were 37% more likely (OR of 1.37; 95% CI, 1.09-1.72). Age at diagnosis was negatively associated, reducing the odds of BCSR by 1% per year increase in age. Compared with white non-Hispanic, Hispanic women were 38% less likely to receive BCSR (OR of 0.62; 95% CI, 0.55-0.71). Married women were 23% more likely to receive BCSR compared with singles (OR of 1.23; 95% CI, 1.09-1.40); women who were separated, divorced, or widowed did not differ significantly from single women. CONCLUSIONS: Distance to radiation therapy facilities may negatively impact the likelihood of BCSR in Florida. Age at diagnosis, insurance type, race/ethnicity, and marital status were associated with BCSR. Future efforts should target the uninsured, Hispanics, the elderly, and the unmarried women to reduce disparities in the administration of BCSR for local breast carcinoma. PMID- 16311988 TI - Properties of excited ketyl radicals of benzophenone analogues affected by the size and electronic character of the aromatic ring systems. AB - The properties of benzophenone ketyl radical analogues with large aromatic ring systems, such as naphthylphenylketone (2), 4-benzoylbiphenyl (3), and bis(biphenyl-4-yl)methanone (4), were investigated in the excited state by using nanosecond-picosecond two-color two-laser flash photolysis. Fluorescence and transient absorption spectra of ketyl radicals of 2-4 in the excited state were observed for the first time. The fluorescence and properties of the excited ketyl radicals were significantly affected by the size and electronic properties of the aromatic ring systems. The reactivity of the ketyl radicals in the excited state with several quenchers was examined and they were found to show reactivity toward N,N-diethylaniline. In addition, for the benzophenone ketyl radical, a unique quenching process of the radical in the excited state by the ground-state parent molecule was found. The factors regulating the fluorescence lifetime of the ketyl radicals in the excited state are discussed quantitatively. PMID- 16311989 TI - Extraction of lanthanides from aqueous solution by using room-temperature ionic liquid and supercritical carbon dioxide in conjunction. AB - For the first time, the study of a three-step extraction system of water/ionic liquid/supercritical CO2 has been performed. Extraction of trivalent lanthanum and europium from an aqueous nitric acid solution to a supercritical CO2 phase via an imidazolium-based ionic liquid phase is demonstrated, and extraction efficiencies higher than 87 % were achieved. The quantitative extraction is obtained by using different fluorinated beta-diketones with and without the addition of tri(n-butyl)phosphate. The complexation phenomenon occurring in the room-temperature ionic-liquid (RTIL) phase was evidenced by using luminescence spectroscopy. PMID- 16311990 TI - A starburst-shaped heterometallic compound incorporating six densely packed gd(3+) ions. AB - The heterotritopic ligand [bpy(DTTA)2]8- has two diethylenediamine-tetraacetate units for selective lanthanide(III) coordination and one bipyridine function for selective Fe(II) coordination. In aqueous solution and in the presence of these metals, the ligand is capable of self-assembly to form a rigid supramolecular metallostar structure, [Fe[Gd2bpy(DTTA)2(H2O)4]3]4-. We report here the physicochemical characterization of the dinuclear complex [Gd2bpy(DTTA)2(H2O)4]2- and the metallostar [Fe[Gd2bpy(DTTA)2(H2O)4]3]4- with regard to potential MRI contrast agent applications. A combination of pH potentiometry and 1H NMR spectroscopy has been used to determine protonation constants for the ligand [bpy(DTTA)2]8- and for the complexes [Fe[bpy(DTTA)2]3]22- and [Y2bpy(DTTA)2]2-. In addition, stability constants have been measured for the dinuclear chelates [M2bpy(DTTA)2]n- formed with M = Gd3+ and Zn2+ (log K(GdL) = 18.2; log K(ZnL) = 18.0; log K(ZnHL) = 3.4). A multiple field, variable-temperature 17O NMR and proton relaxivity study on [Gd2bpy(DTTA)2(H2O)4]2- and [Fe[Gd2bpy(DTTA)2(H2O)4]3](4-) yielded the parameters for water exchange and the rotational dynamics. The 17O chemical shifts are indicative of bishydration of the lanthanide ion. The exchange rates of the two inner-sphere water molecules are very similar in the dinuclear [Gd2bpy(DTTA)2(H2O)(4)]2- and in the metallostar (k(ex)298 = 8.1 +/- 0.3 x 10(6) and 7.4 +/- 0.2 x 10(6) s(-1), respectively), and are comparable to k(ex)298 for similar Gd(III) poly(amino carboxylates). The rotational dynamics of the metallostar has been described by means of the Lipari-Szabo approach, which involves separating global and local motions. The difference between the local and global rotational correlation times, tau(lO)298 = 190 +/- 15 ps and tau(gO)298 = 930 +/- 50 ps, respectively, shows that the metallostar is not completely rigid. However, the relatively high value of S2 = 0.60 +/- 0.04, describing the restriction of the local motions with regard to the global one, points to a limited flexibility compared with previously reported macromolecules such as dendrimers. As a result of the two inner-sphere water molecules, with their near-optimal exchange rate, and the limited flexibility, the metallostar has a remarkable molar proton relaxivity, particularly at high magnetic fields (r1 = 33.2 and 16.4 mM(-1) s(-1) at 60 and 200 MHz, respectively, at 25 degrees C). It packs six efficiently relaxing Gd(III) ions into a small molecular space, which leads, to the best of our knowledge, to the highest relaxivity per molecular mass ever reported for a Gd(III) complex. The [bpy(DTTA)2]8- ligand is also a prime candidate as a terminal ligand for constructing larger sized, Fe(II) (or Ru(II))-based metallostars or metallodendrimers loaded with Gd(III) on the surface. PMID- 16311991 TI - A new entry to doubly N-confused [26]hexaphyrins(1.1.1.1.1.1) from normal [26]hexaphyrins(1.1.1.1.1.1) through an unprecedented double pyrrolic rearrangement. AB - Treatment of hexakis(pentafluorophenyl)-substituted [26]hexaphyrin(1.1.1.1.1.1) (1) with CuCl in the presence of pyridine and molecular oxygen led to formation of doubly N-confused [26]hexaphyrin(1.1.1.1.1.1) 2 in a moderate yield through an unprecedented double pyrrolic rearrangement. Macrocycle 2 has been shown to serve as an effective bis-metal-coordinating ligand and exhibits attractive optical properties such as a sharp Soret-like band at 566 nm and low-energy fluorescence at 1058 nm. PMID- 16311992 TI - Synthesis of semiconducting thin films with nanometer-scale periodicity by solution-phase coassembly of zintl clusters with surfactants. PMID- 16311993 TI - Complexation-induced translational isomerism: shuttling through stepwise competitive binding. PMID- 16311994 TI - Total synthesis, NMR solution structure, and binding model of the potent histone deacetylase inhibitor FR235222. PMID- 16311995 TI - High-pressure probing of a changeover in the charge-transfer mechanism for intact cytochrome c at gold/self-assembled monolayer junctions. PMID- 16311996 TI - Enantiomerically pure copper(II) Cubanes [Cu4L2(OMe)4] from chiral bis-1,3 diketones H2L through diastereoselective self-assembly. PMID- 16311997 TI - Sustained and therapeutic levels of human factor IX in hemophilia B mice implanted with microcapsules: key role of encapsulated cells. AB - BACKGROUND: A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein was explored in this study. In order to prevent immune rejection of the delivered cells, they were enclosed in non-antigenic biocompatible alginate microcapsules prior to being implanted intraperitoneally into mice. We have shown that encapsulated C2C12 myoblasts can temporarily deliver therapeutic levels of factor IX (FIX) in mice, but the C2C12 myoblasts elicited an immune response to FIX. In this study we report the use of mouse fetal G8 myoblasts secreting hFIX in hemophilia mice. METHODS: Mouse G8 myoblasts were transduced with MFG-FIX vector. A pool of recombinant G8 myoblasts secreting approximately 1500 ng hFIX/10(6) cells/24 h in vitro were enclosed in biocompatible alginate microcapsules and implanted intraperitoneally into immunocompetent C57BL/6 and hemophilic mice. RESULTS: Circulating levels of hFIX in treated mice reached approximately 400 ng/ml for at least 120 days (end of experiment). Interestingly, mice treated with encapsulated G8 myoblasts did not develop anti-hFIX antibodies. Activated partial thromboplastin time (APTT) of plasmas obtained from treated hemophilic mice was reduced from 107 to 82 sec on day 60 post-treatment, and whole blood clotting time (WBCT) was also corrected from 7-9 min before treatment to 3-5 min following microcapsule implantation. Further, mice were protected against bleeding following major trauma. Thus, the FIX delivery in vivo was biologically active. CONCLUSIONS: Our findings suggest that the type of cells encapsulated play a key role in the generation of immune responses against the transgene. Further, a judicious selection of encapsulated cells is critical for achieving sustained gene expression. Our findings support the feasibility of encapsulated G8 myoblasts as a gene therapy approach for hemophilia B. PMID- 16311998 TI - Ru nanoparticles immobilized on montmorillonite by ionic liquids: a highly efficient heterogeneous catalyst for the hydrogenation of benzene. PMID- 16311999 TI - Spectrally resolved magnetic resonance imaging of a xenon biosensor. PMID- 16312000 TI - Addressing the metal centers of [2x2] CoII4 grid-type complexes by STM/STS. PMID- 16312001 TI - Folding a de novo designed peptide into an alpha-helix through hydrophobic binding by a bowl-shaped host. PMID- 16312002 TI - Template-free synthesis and assembly of single-crystalline tungsten oxide nanowires and their gas-sensing properties. PMID- 16312003 TI - Lanthanide-transition-metal sandwich framework comprising {Cu3} cluster pillars and layered networks of {Er36} wheels. PMID- 16312004 TI - Ferrocene-mediated proton-coupled electron transfer in a series of ferrocifen type breast-cancer drug candidates. PMID- 16312006 TI - Fivefold coordination of a CuII-aqua ion: a supramolecular sandwich consisting of two crown ether molecules and a trigonal-bipyramidal [Cu(H2O)5]2+ Complex. PMID- 16312005 TI - Enantioselective PhSe-group-transfer tandem radical cyclization reactions catalyzed by a chiral Lewis acid. PMID- 16312007 TI - The C-H...pi hydrogen bond in the benzene-trifluoromethane adduct: a rotational study. PMID- 16312008 TI - Copper(I)-directed formation of a cyclic pseudorotaxane tetramer and its trimeric homologue. PMID- 16312009 TI - Gamma-silyl group effect in hydroalumination and carbolithiation of propargylic alcohols. PMID- 16312010 TI - Short and efficient approach towards macrocyclic lactones based on a Sonogashira reaction. AB - Polyketide-derived macrolactones like zearalenone (1), zearalane (2) or curvularin (3) display a wide range of interesting pharmacological activities. Here, we present a short and efficient approach towards this class of natural products by a combination of the Sonogashira and Mitsunobu reactions. The resulting lactone 9 was tested against human cancer cell lines at the NCI. PMID- 16312011 TI - Pharmacokinetics of diclofenac in rat model of diabetes mellitus induced by alloxan or steptozotocin. AB - The pharmacokinetics of diclofenac were compared after intravenous and oral administration at a dose of 5 mg/kg in a rat model of diabetes mellitus induced by alloxan (DMIA) or streptozotocin (DMIS), and their respective control rats. Diclofenac was reported to be metabolized via the hepatic microsomal cytochrome P450 (CYP) 2C11 in male rats. The expression and mRNA level of CYP2C11 decreased in rat models of DMIA and DMIS. Hence, the time-averaged nonrenal clearance (Clnr) of diclofenac was expected to be slower in a rat model of diabetes. As expected, after intravenous administration, the Clnr values of diclofenac were significantly slower in rat models of DMIA (11.3 versus 13.6 ml/min/kg) and DMIS (8.06 versus 15.2 ml/min/kg) than those in control rats. As a result, the total area under the plasma concentration-time curve from time zero to time infinity (AUC) values were significantly greater in rat models of DMIA (435 versus 367 microg min/ml) and DMIS (540 versus 329 microg min/ml). However, after oral administration, the AUC from time zero to the last measured time, 12 h, in plasma (AUC0-12 h) values were comparable between the rat models of DMIA and DMIS and their control rats, and this could be due to changes in the first-pass effect of diclofenac and was not due to a decrease in the absorption of diclofenac in the rat models of diabetes. PMID- 16312013 TI - A longitudinal study of ethanol and acetaldehyde in the exhaled breath of healthy volunteers using selected-ion flow-tube mass spectrometry. AB - Selected-ion flow-tube mass spectrometry (SIFT-MS) has been used to monitor the volatile compounds in the exhaled breath of 30 volunteers (19 male, 11 female) over a 6-month period. Volunteers provided breath samples each week between 8:45 and 13:00 (before lunch), and the concentrations of several trace compounds were obtained. In this paper the focus is on ethanol and acetaldehyde, which were simultaneously quantified by SIFT-MS using H3O+ precursor ions. The mean ethanol level for all samples was 196 parts-per-billion (ppb) with a standard deviation of 244 ppb, and the range of values for breath samples analysed is 0 to 1663 ppb. The mean acetaldehyde level for all samples was 24 ppb with a standard deviation of 17 ppb, and the range of values for breath samples analysed is 0 to 104 ppb. Background (ambient air) levels of ethanol were around 50 ppb, whereas any background acetaldehyde was usually undetectable. Increased ethanol levels were observed if sweet drink/food had been consumed within the 2 h prior to providing the breath samples, but no increase was apparent when alcohol had been consumed the previous evening. The measured endogenous breath ethanol and acetaldehyde levels were not correlated. These data relating to healthy individuals are a prelude to using breath analysis for clinical diagnosis, for example, the recognition of bacterial overload in the gut (ethanol) or the possibly of detecting tumours in the body (acetaldehyde). PMID- 16312014 TI - Seroprevalence study of Tick-borne encephalitis, Borrelia burgdorferi, Dengue and Toscana virus in Turin Province. AB - Tick borne encephalitis virus (TBEV) is present in some European countries and it is transmitted by a tick bite. Ixodes ricinus is the main vector of the infection in Italy, where fortunately clinical neurological manifestations, typical of the more serious phase of the disease, are very rarely observed. This behaviour is different from other endemic Euroasiatic areas where numerous cases of encephalitis are described. However TBE transmission has not been widely investigated in Italy and available epidemiological data have been obtained only by studies performed in Central and Northern Regions of the country. In addition seroepidemiological researches were made prevalently on subjects at high risk of tick bite, such as hunters or forest guards from Trentin and Central Italy. No precise information about TBE virus diffusion was available in the Piedmont before our investigations. We found that hunters and wild boar breeders seem to be particularly exposed to the risk of TBE virus infection in Turin Province and in particular in the Susa valley, although no neurological involvement was observed in our population. In particular a seroprevalence of about 5% was detected by the use of purified antigens ELISA test, amongst the subjects at high risk of tick bite. Moreover low risk individuals showed a seroprevalence of below 2%. In addition a parallel seroepidemiological study was performed in Turin Province for Borrelia burgdorferi, the aetiological agent of Lyme disease, also transmitted by tick bite (e.g. Ixodes ricinus), for Dengue and Toscana (TOS) arboviruses, respectively transmitted by Aedes mosquitoes and phlebotomes. Data reported here demonstrate only a sporadic presence in our population of antibodies against Borrelia and Dengue infection. Moreover using an ELISA test performed with viral purified nucleoprotein, we reported a total percentage of about 3% of subjects positive for TOSV. PMID- 16312015 TI - Base-pairing potential identified by in vitro selection predicts the kinked RNA backbone observed in the crystal structure of the alfalfa mosaic virus RNA-coat protein complex. AB - The three-dimensional structure of the 3' terminus of alfalfa mosaic virus RNA in complex with an amino-terminal coat protein peptide revealed an unusual RNA fold with inter-AUGC basepairing stabilized by key arginine residues (Guogas, et al., 2004). To probe viral RNA interactions with the full-length coat protein, we have used in vitro genetic selection to characterize potential folding patterns among RNAs isolated from a complex randomized pool. Nitrocellulose filter retention, electrophoretic mobility bandshift analysis, and hydroxyl radical footprinting techniques were used to define binding affinities and to localize the potential RNA-protein interaction sites. Minimized binding sites were identified that included both the randomized domain and a portion of the constant regions of the selected RNAs. The selected RNAs, identified by their ability to bind full-length coat protein, have the potential to form the same unusual inter-AUGC Watson-Crick base pairs observed in the crystal structure, although the primary sequences diverge from the wild-type RNA. A constant feature of both the wild-type RNA and the selected RNAs is a G ribonucleotide in the third position of an AUGC-like repeat. Competitive binding assays showed that substituting adenosine for the constant guanosine in either the wild-type or selected RNAs impaired coat protein binding. These data suggest that the interactions observed in the RNA-peptide structure are likely recapitulated when the full-length protein binds. Further, the results underscore the power of in vitro genetic selection for probing RNA protein structure and function. PMID- 16312016 TI - Prediction of spectroscopic constants for diatomic molecules in the ground and excited states using time-dependent density functional theory. AB - Spectroscopic constants of the ground and next seven low-lying excited states of diatomic molecules CO, N2, P2, and ScF were computed using the density functional theory SAOP/ATZP model, in conjunction with time-dependent density functional theory (TD-DFT) and a recently developed Slater type basis set, ATZP. Spectroscopic constants, including the equilibrium distances r(e), harmonic vibrational frequency omega(e), vibrational anharmonicity omega(e)x(e), rotational constant B(e), centrifugal distortion constant D(e), the vibration rotation interaction constant alpha(e), and the vibrational zero-point energy E(n)0 were generated in an effort to establish a reliable database for electron spectroscopy. By comparison with experimental values and a similar model with an established larger Slater-type basis set, et-QZ3P-xD, it was found that this model provides reliably accurate results at reduced computational costs, for both the ground and excited states of the molecules. The over all errors of all eight lowest lying electronic states of the molecules under study using the effective basis set are r(e)(+/-4%), omega(e)(+/-5% mostly without exceeding +/-20%), omega(e)x(e)(+/-5% mostly without exceeding 20%, much more accurate than a previous study on this constant of +/-30%), B(e)(+/-8%), D(e)(+/-10%), alpha(e)(+/-10%), and E(n)0(+/-10%). The accuracy obtained using the ATZP basis set is very competitive to the larger et-QZ3P-xD basis set in particular in the ground electronic states. The overall errors in r(e), omega(e)x(e), and alpha(e) in the ground states were given by +/-0.7, +/-10.1, and +/-8.4%, respectively, using the efficient ATZP basis set, which is competitive to the errors of +/-0.5, +/-9.2, and +/-9.1%, respectively for those constants using the larger et-QZ3P-xD basis set. The latter basis set, however, needs approximately four times of the CPU time on the National Supercomputing Facilities (Australia). Due to the efficiency of the model (TD-DFT, SAOP and ATZP), it will be readily applied to study larger molecular systems. PMID- 16312017 TI - Reaction of nitrones with silyl ketene acetals: a DFT study. AB - Theoretical calculations at the DFT level indicate that the reaction of a nitrone with a silyl ketene acetal proceeds, contrary to previous suggestions, through a classical 1,3-dipolar cycloaddition, followed by a silyl group transfer step to give the open-chain product. Introduction of a diffuse basis set is necessary to describe properly nitrones. The influence of a Lewis acid has been studied. PMID- 16312018 TI - Revisiting the geometry of nd10 (n+1)s0 [M(H2O)]p+ complexes using four-component relativistic DFT calculations and scalar relativistic correlated CSOV energy decompositions (M(p+) = Cu+, Zn2+, Ag+, Cd2+, Au+, Hg2+). AB - Hartree-Fock and DFT (B3LYP) nonrelativistic (scalar relativistic pseudopotentials for the metallic cation) and relativistic (molecular four component approach coupled to an all-electron basis set) calculations are performed on a series of six nd10 (n+1)s0 [M(H2O)]p+ complexes to investigate their geometry, either planar C2v or nonplanar C(s). These complexes are, formally, entities originating from the complexation of a water molecule to a metallic cation: in the present study, no internal reorganization has been found, which ensures that the complexes can be regarded as a water molecule interacting with a metallic cation. For [Au(H2O)]+ and [Hg(H2O)]2+, it is observed that both electronic correlation and relativistic effects are required to recover the C(s) structures predicted by the four-component relativistic all-electron DFT calculations. However, including the zero-point energy corrections makes these shallow C(s) minima vanish and the systems become floppy. In all other systems, namely [Cu(H2O)]+, [Zn(H2O)]2+, [Ag(H2O)]+, and [Cd(H2O)]2+, all calculations predict a C2v geometry arising from especially flat potential energy surfaces related to the out-of-plane wagging vibration mode. In all cases, our computations point to the quasi-perfect transferability of the atomic pseudopotentials considered toward the molecular species investigated. A rationalization of the shape of the wagging potential energy surfaces (i.e., single well vs. double well) is proposed based on the Constrained Space Orbital Variation decompositions of the complexation energies. Any way of stabilizing the lowest unoccupied orbital of the metallic cation is expected to favor charge transfer (from the highest occupied orbital(s) of the water ligand), covalence, and, consequently, C(s) structures. The CSOV complexation energy decompositions unambiguously reveal that such stabilizations are achieved by means of relativistic effects for [Au(H2O)]+, and, to a lesser extent, for [Hg(H2O)]2+. Such analyses allow to numerically quantify the rule of thumb known for Au+ which, once again, appears as a better archetype of a relativistic cation than Hg2+. This observation is reinforced due to the especially high contribution of the nonadditive correlation/relativity terms to the total complexation energy of [Au(H2O)]+. PMID- 16312019 TI - Monensin A methyl ester complexes with Li+, Na+, and K+ cations studied by ESI MS, 1H- and 13C-NMR, FTIR, as well as PM5 semiempirical method. AB - Monensin A methyl ester (MON1) was synthesized by a new method and its ability to form complexes with Li+, Na+, and K+ cations was studied by electrospray ionization-mass spectroscopy (ESI-MS), 1H and 13C nuclear magnetic resonance (NMR), Fourier transform infrared (FTIR), and PM5 semiempirical methods. It is shown that MON1 with monovalent metal cations forms stable complexes of 1:1 stoichiometry. The structures of the complexes are stabilized by intramolecular hydrogen bonds in which the OH groups are always involved. In the structure of MON1, the oxygen atom of the C=O ester group is involved in very weak bifurcated intramolecular hydrogen bonds with two hydroxyl groups, whereas in the complexes of MON1 with monovalent metal cations the C=O ester group is not engaged in any intramolecular hydrogen bonds. Furthermore, it is demonstrated that the strongest intramolecular hydrogen bonds are formed within the MON1-Li+ complex structure. The structures of the MON1 and its complexes with Li+, Na+, and K+ cations are visualized and discussed in detail. PMID- 16312020 TI - Interaction between complementary liposomes: a process leading to multicompartment systems formation. AB - Interaction of complementary liposomes induces a series of processes, involving reorganization of their membrane lipids, which lead to the formation of large aggregates. In several cases these aggregates exhibit multicompartment structures and only primitively mimic, in some aspects at least, the multicompartmental features of cells. Similar multicompartment structures were repeatedly obtained following the interaction of a diversity of complementary liposomal pairs. Thus, a working hypothesis is proposed, according to which, molecular recognition of liposomes induces the formation of multicompartment structures. PMID- 16312021 TI - Characterization of prostate-specific antigen binding peptides selected by phage display technology. AB - Prostate-specific antigen (PSA) is an important marker for the diagnosis and management of prostate cancer. Free PSA has been shown to be more extensively cleaved in sera from benign prostatic hyperplasia patients than in sera from prostate cancer patients. Moreover, the presence of enzymatically activatable PSA was characterized previously in sera from patients with prostate cancer by the use of the specific anti-free PSA monoclonal antibody (mAb) 5D3D11. As an attempt to obtain ligands for the specific recognition of different PSA forms including active PSA, phage-displayed linear and cyclic peptide libraries were screened with PSA coated directly into microplate wells or presented by two different anti total PSA mAbs. Four different phage clones were selected for their ability to recognize PSA and the inserted peptides were produced as synthetic peptides. These peptides were found to capture and to detect specifically free PSA, even in complex biological media such as sera or tumour cell culture supernatants. Alanine scanning of peptide sequences showed the involvement of aromatic and hydrophobic residues in the interaction of the peptides with PSA whereas Spotscan analysis of overlapping peptides covering the PSA sequence identified a peptide binding to the kallikrein loop at residues 82-87, suggesting that the peptides could recognize a non-clipped form of PSA. Moreover, the PSA-specific peptides enhance the enzymatic activity of PSA immobilized into microplate wells whereas the capture of PSA by the peptides inhibited totally its enzymatic activity while the peptide binding to PSA had no effect in solution. These PSA-specific peptides could be potential tools for the recognition of PSA forms more specifically associated to prostate cancer. PMID- 16312022 TI - Intercalation of daunomycin into d(CG)4 oligomer duplex containing G x T mismatches by vibrational circular dichroism and infrared absorption spectroscopy. AB - The vibrational circular dichroism (VCD) and infrared absorption (IR) spectra of the mismatched octamer oligonucleotides d(CGTGCGCG)(2) (CGT) and d(CGCGTGCG)(2) (CGC) and their complexes with the antitumor drug daunomycin were measured in D(2)O, interpreted, and compared to the octamer d(CGCGCGCG)(2) (CG). The IR spectra of the mismatched octamers in the carbonyl-stretching region are similar to those of the parent CG, whereas the VCD spectra differ in several respects between each other. The main VCD feature due to carbonyl stretching is informative for the mismatches and CG. Vibrational modes in the sugar-phosphate region remain essentially unchanged especially for PO(2) (-) symmetric stretching. Differences between the free and complexed mismatch octamers occurred mainly in the carbonyl-stretching region (1,700-1,600 cm(-1)). The absorption intensity of the C==O peak of G is more prominent for CGC than CGT and resembles CG in this respect. The detailed composition of this doublet is clearly visible, indicating the geometric rearrangement of the base pairs in the presence of the mismatch and upon forming the daunomycin complex. PMID- 16312023 TI - Post-consumer contamination in high-density polyethylene (HDPE) milk bottles and the design of a bottle-to-bottle recycling process. AB - Six hundred conventional recycled HDPE flake samples, which were recollected and sorted in the UK, were screened for post-consumer contamination levels. Each analysed sample consisted of 40-50 individual flakes so that the amount of analysed individual containers was in the range 24,000-30,000 post-consumer milk bottles. Predominant contaminants in hot-washed flake samples were unsaturated oligomers, which can be also be found in virgin high-density polyethylene (HDPE) pellet samples used for milk bottle production. In addition, the flavour compound limonene, the degradation product of antioxidant additives di-tert-butylphenol and low amounts of saturated oligomers were found in higher concentrations in the post-consumer samples in comparison with virgin HDPE. However, the overall concentrations in post-consumer recycled samples were similar to or lower than concentration ranges in comparison with virgin HDPE. Contamination with other HDPE untypical compounds was rare and was in most cases related to non-milk bottles, which are <2.1% of the input material of the recycling process. The maximum concentration found in one sample of 1 g was estimated as 130 mg kg(-1), which corresponds to a contamination of 5200-6500 mg kg(-1) in the individual bottle. The recycling process investigated was based on an efficient sorting process, a hot-washing of the ground bottles, and a further deep-cleaning of the flakes with high temperatures and vacuum. Based on the fact that the contamination levels of post-consumer flake samples are similar to virgin HDPE and on the high cleaning efficiency of the super-clean recycling process especially for highly volatile compounds, the recycling process investigated is suitable for recycled post-consumer HDPE bottles for direct food-contact applications. However, hand-picking after automatically sorting is recommended to decrease the amount of non-milk bottles. The conclusions for suitability are valid, provided that the migration testing of recyclate contains milk bottles up to 100% and that both shelf-life testing and sensorial testing of the products are successful, which are topics of further investigations. PMID- 16312024 TI - Surgical treatment of rectal prolapse ischemia. PMID- 16312025 TI - Abdominopelvic mass secondary to Actinomyces israelii mimicking cancer: report of two cases. PMID- 16312026 TI - Stomal plugs: a novel treatment option for stomal stenosis. PMID- 16312027 TI - Electrocoagulation: an alternative treatment for rectal cancer. PMID- 16312028 TI - Micronutrients and neutraceutics and functional foods. PMID- 16312029 TI - Medicare launches Physician Voluntary Reporting Program. PMID- 16312030 TI - Risk calculation of fetal chromosomal defects in the first trimester. PMID- 16312031 TI - [Comments on the article "The nasal bone in first trimester screenings" by P. Schmidt, I Staboulidou and C. Sohn in "Gynecology", August 2005]]. PMID- 16312033 TI - BEIR VII-2. PMID- 16312032 TI - Further comment on 'Personal photon dosemeter trial--Devonport Royal Dockyard'. PMID- 16312034 TI - Low doses of radiation linked to small increase in cancer risk. PMID- 16312035 TI - Low levels of ionising radiation may cause harm. PMID- 16312036 TI - COMARE 10th report: the incidence of childhood cancer around nuclear installations in Great Britain. PMID- 16312038 TI - The Article 31 Group of experts' Working Party on education and training. PMID- 16312037 TI - Childhood cancer in relation to distance from high-voltage power lines in England and Wales: a case-control study. PMID- 16312039 TI - Conquering schistosomiasis in China: the long march. AB - The last half-century of schistosomiasis control activities in China have brought down the overall prevalence of human infection with Schistosoma japonicum to less than 10% of the level initially documented in the mid 1950s. Importantly, this reduction is not only, or even mainly, due to the advent of praziquantel in the 1970s and its subsequent dramatic fall in price. Instead, it is the result of a sustained, multifaceted national strategy, adapted to different eco epidemiological settings, which has been versatile enough to permit subtle adjustments over time as the nature of the challenge changed. Consequently, prevalence has been falling relatively smoothly over the whole period rather than suddenly dropping when mass chemotherapy became feasible. Thus, early recognition of the huge public health and economic significance of the disease, and the corresponding political will to do something about it,underpinned this success. In addition, intersectoral collaboration and community participation played important roles in forming a sustained commitment to a working control strategy based on local resources. The unfolding story is presented from the early years' strong focus on snail control, by means of environmental management, to the last period of praziquantel-based morbidity control carried out under the 10-year World Bank Loan Project (WBLP). An important legacy of the WBLP is the understanding that a research component would sustain control measures and enable future progress. We are now witnessing the payoffs of this forward thinking in the form of a new promising class of drugs, improved diagnostics, and budding vaccine development in addition to novel ways of disease risk prediction and transmission control using satellite-based remote sensing. Different aspects of social and economic approaches are also covered and the importance of health promotion and education is emphasized. Issuing from the review is a set of recommendations, which might further consolidate current control activities, with the ultimate aim to eliminate schistosomiasis from the Chinese mainland. PMID- 16312041 TI - [Study on thinking of clinical scientific research of acupuncture and moxibustion]. AB - Under guiding of correct thinking of scientific research, development of clinical scientific research of acupuncture and moxibustion is of important significance for promoting academic progresses of acupuncture and moxibustion, increasing clinical therapeutic effect of acupuncture and moxibustion. This paper probes thinking of clinical scientific research of acupuncture and moxibustion and puts forward the guiding principles, i.e. testing and verifying therapeutic effect, searching for laws, expounding mechanisms and guiding clinical practice. PMID- 16312042 TI - Local delivery of doxorubicin for the treatment of malignant brain tumors in rats. AB - BACKGROUND: Local delivery of carmustine (BCNU) via biodegradable polymers has been shown to improve survival in patients with glioblastoma multiforme (GBM). In the current study, we hypothesized that local delivery of an anthracycline antibiotic, doxorubicin (DOX), might act to improve the survival of animals bearing experimental intracranial glioma. MATERIALS AND METHODS: Polyanhydride polymers (PCPP-SA), containing either 3% or 5% ADR by weight, were prepared using the mix-melt method. Forty male Fisher 344 rats received an intracranial challenge with a lethal dose of 9L gliosarcoma cells. Five days later, they received DOX or blank polymer. There were a total of four treatment groups: (1) blank polymer; (2) 3% DOX polymer; (3) 5% DOX polymer, and (4) control group with no polymer. RESULTS: Compared to control animals treated with no polymers or blank polymer, animals receiving DOX had significantly extended survival. The median survival for the control group was 21 days vs. 34 days (p < 0.01) for the 3% DOX group and 45 days (p < 0.0001) for the 5% DOX group. CONCLUSION: Doxorubicin, when delivered locally, is an effective monotherapeutic agent against experimental intracranial glioma. PMID- 16312043 TI - Hyperbaric oxygen inhibits benign and malignant human mammary epithelial cell proliferation. AB - BACKGROUND: Hyperbaric oxygenation (HBO) therapy is the administration of 100% inhaled oxygen to patients at increased atmospheric pressure. MATERIALS AND METHODS: We used an in vitro model to examine the effects of HBO on mammary cell proliferation. Normal mammary epithelia, primary tumor and metastatic tumor cells derived from the same patient and immortalized by transfection with the human papilloma virus E6 oncogene, as well as the MCF7 human mammary adenocarcinoma cell line, were studied. RESULTS: HBO (97.9% O2, 2.1% CO2, 2.4 atmospheres absolute) inhibited the proliferation of all 4 cell types as measured by light microscopy, [3H]thymidine uptake, a tetrazolium-based colorimetric assay and a clonogenicity assay. The anti-proliferative effect of HBO was time-dependent (p < 0.01 for all 4 cell types). Hyperoxia alone (95% O2, 5% CO2, 1 atmosphere absolute) and increased atmospheric pressure alone (8.75% O2, 2.1% CO2, 2.4 atmospheres absolute) also inhibited proliferation, but their effects were not as profound as HBO (p < 0.01 when either hyperoxia or increased pressure was compared to HBO for all 4 cell types). HBO enhanced the anti-proliferative effects of melphalan (p < 0.05), gemcitabine (p < 0.001) and paclitaxel (p < 0.001). The clonogenicity assay demonstrated that the effects of HBO were still evident 2 weeks after the exposure (p < 0.01 for all 4 cell types). Experiments using Hoechst-propidium iodide or annexin V-propidium iodide staining showed no HBO-induced increases in necrosis or apoptosis. CONCLUSION: HBO inhibits benign and malignant mammary epithelial cell proliferation, but does not enhance cell death. PMID- 16312044 TI - CIITA transformation rescues the apoptotic function of MHC class II in melanoma cells. AB - BACKGROUND: Cell-surface major histocompatibility (MHC) class II molecules contribute to a molecular, intercellular complex that stimulates T-cells. MHC class II molecules also activate signaling pathways leading to apoptosis. Lack of CIITA, a co-activator of the MHC class II gene promoter, is responsible for lack of MHC class II on most of the MHC class II-negative melanoma cell lines. MATERIALS AND METHODS: We rescued CIITA and MHC class II expression in melanoma cell lines by stable transformation with a CIITA expression vector and assayed for MHC class II-mediated apoptosis. RESULTS: Reconstitution of CIITA in multiple CIITA-negative melanoma lines restores the apoptotic function of melanoma MHC class II. CONCLUSION: Lack of MHC class II transcription, rather than defects in MHC class II protein function or defects in the other components of the MHC class II-stimulated apoptotic pathway, prevents MHC class II-mediated apoptosis in melanoma cells. PMID- 16312045 TI - Distant metastasis from subcutaneously grown E0771 medullary breast adenocarcinoma. AB - BACKGROUND: Breast cancer treatments are most effective when initiated early, with very poor efficacy against metastatic disease. In seeking a readily metastasizing mouse breast cancer model to facilitate the search for effective therapies, E0771 medullary adenocarcinomas implanted subcutaneously in syngeneic C57BL/6 mice were studied. MATERIALS AND METHODS: Standard pathological, histological and immunological methodologies were used. RESULTS: The aggressive estrogen receptor-positive tumor invaded locally into the peritoneal cavity in 56% of mice, as well as metastasizing to the lungs in 52% of mice. The metastasis was a spontaneous event and immunosuppression was seen (e.g. generation of lyphokine activated killer cells and allogeneic cytotoxic T lymphocytes cytolytic activities ex vivo were suppressed). Other pathological events noted as the tumor progressed were: bloody ascites (56%) and shock (72%), both attributed to local (peritoneal) tumor invasion. CONCLUSION: The E0771 metastatic breast cancer model, which mimics the human disease, should be useful in testing new treatments against this disease and/or in examining the metastatic process. PMID- 16312046 TI - Effects of human breast stromal cells on conjugated linoleic acid (CLA) modulated vascular endothelial growth factor-A (VEGF-A) expression in MCF-7 cells. AB - BACKGROUND: Conjugated linoleic acid (CLA), a naturally occurring compound found in ruminant dairy and beef products, has been shown to possess anti-cancer ability in vivo and in vitro. There are several CLA isomers in ruminant-produced food products, among which t10,c12-CLA and c9,t11-CLA are most potent. Vascular endothelial growth factor-A (VEGF-A) has been implicated as an angiogenesis activating cytokine. Our previous results indicated that CLA induced suppression of VEGF-A in MCF-7 cells, which may be one of CLA's anticancer mechanisms. MATERIALS AND METHODS: The effects of 10,c12-CLA and c9,t11-CLA on VEGF-A mRNA and protein expression in MCF-7 cells, which were co-cultured with human breast stromal cells isolated from breast tissues of surgical specimens of mammoplasty and breast cancer patients, were detected by RT-PCR and Western blot analysis. RESULTS: VEGF-A mRNA and protein expressions were significantly (p < 0.05) elevated in co-cultured MCF-7 cells in comparison with cultured MCF-7 cells alone. Normal human breast stromal cells contribute greater effects in increasing VEGF-A protein expression in MCF-7 cells. Both t10,c12-CLA and c9,t11-CLA significantly (p < 0.05) decreased VEGF-A mRNA and protein levels in co-cultured MCF-7 cells. t10,c12-CLA appeared to be the more active isomer than c9, t11-CLA. CONCLUSION: The results indicate that dietary CLA might serve as a chemo therapeutic agent in human breast cancers by down-regulating VEGF-A expression. PMID- 16312047 TI - Effects of age on growth and ERbeta mRNA expression of canine prostatic cells. AB - BACKGROUND: Benign prostatic hyperplasia (BPH) is an age-dependent prostatic disease in human males and dogs. The prostatic stromal estrogen level of health control and BPH patients increases significantly with age, while the dihydrotestosterone (DHT) level is not connected with age. Moreover, experimentally estrogens have induced BPH in the presence of androgens. Our aim was to investigate the effects of age on the proliferation and estrogen receptor beta (ER/3) mRNA of canine prostatic epithelial and stromal cells. MATERIALS AND METHODS: Epithelial and stromal cells were isolated from canine prostatic tissues. The proliferation of these cell types from dogs of different ages was assessed by thymidine incorporation assay, while the expression and identification of ERbeta mRNA were performed by RT-PCR and DNA sequence. RESULTS: Prostatic epithelial cells isolated from 1-year-old dogs exhibited a greater proliferative activity than those of 4-year-old dogs. In contrast, the prostatic stromal cells from 4-year-old dogs proliferated more rapidly than the cells from 1-year-old dogs. ERbeta mRNA expression was detected in the canine prostatic epithelial and stromal cells, decreasing with age. The partial DNA sequence showed that the canine ERbeta sequence shares 90.0%, 87.0% and 83.0% of its nucleotide homology with human, rat and mouse ERbeta, respectively. CONCLUSION: The decrease in the expression of ERbeta in prostatic cells with age reduces its negative control over the androgen receptor, associated with the overgrowth of canine prostatic stromal cells, which further induces the development of canine BPH. PMID- 16312048 TI - The immune epitope database and analysis resource: from vision to blueprint. PMID- 16312049 TI - Revealing the structure and dynamics of cis-regulation using heterogeneous, genome-wide, multi-species data. PMID- 16312050 TI - Complex systems biology: exploring universal statistical and dynamical features in cellular processes. PMID- 16312051 TI - Oligo- and poly-nucleotides: 50 years of chemical synthesis. AB - It is fifty years since the first chemical synthesis of a dinucleoside phosphate and a dinucleotide with natural 3'-->5'-internucleotide linkages was reported. The main developments in the methodology of oligo- and poly-nucleotide synthesis that have taken place since are described. PMID- 16312052 TI - Conversation with Shanthi Ranganathan. PMID- 16312053 TI - Federal enforcement in group and individual health insurance markets. Final rule. AB - This rule makes final an interim final rule that details procedures we use for enforcing title XXVII of the Public Health Service Act as added by the Health Insurance Portability and Accountability Act of 1996, and as amended by the Mental Health Parity Act of 1996, the Newborns' and Mothers' Health Protection Act of 1996, and the Women's Health and Cancer Rights Act of 1998. Specifically, we are responsible for enforcing title XXVII requirements in States that do not enact the legislation necessary to enforce those requirements, or otherwise fail to substantially enforce the requirements. We are also responsible for taking enforcement actions against non-Federal governmental plans. The regulation describes the process we use in both enforcement contexts. This final rule deletes an appendix to the interim rule that listed examples of violations of title XXVII and corrects the description of a cross-reference, but makes no substantive changes to the interim final rule. PMID- 16312054 TI - Comments on time-efficient technique for improved tissue contrast in neuroimaging. PMID- 16312055 TI - Nuclear medicine affected by Energy Policy Act of 2005. PMID- 16312056 TI - Lung cancer awareness month. PMID- 16312057 TI - Graduated pressure regimen for external counterpulsation (ECP) for patients with heart failure. PMID- 16312058 TI - Economy chased medicine: health under the provision of economy -- chance or threat for CAM? PMID- 16312059 TI - WHO launches global patient safety challenge; issues guidelines on hand hygiene in health care. PMID- 16312060 TI - "Stop the global epidemic of chronic disease"--new report, preventing chronic diseases: a vital investment estimates hundreds of billions of dollars at stake. PMID- 16312061 TI - Global efforts to prevent violence: World Health Organization reports significant progress. PMID- 16312062 TI - Hepatitis A associated with green onions. PMID- 16312063 TI - Laurie A. Kruzich, MS, RD, wins Huddleston Award. PMID- 16312064 TI - Nomogram for exercise capacity in women. PMID- 16312065 TI - Nomogram for exercise capacity in women. PMID- 16312066 TI - Penny Kris-Etherton, PhD, RD, wins Monsen Award. PMID- 16312067 TI - Sweat the small stuff: patients do! PMID- 16312069 TI - [Risk management--adverse effects of drugs. Outline of drug reactions and safety policy]. PMID- 16312068 TI - United Ostomy Association to cease operations. PMID- 16312071 TI - Action called for on infectious diseases. PMID- 16312070 TI - Care homes failing to address continence. PMID- 16312072 TI - Nurses left out of DoH consultation. PMID- 16312073 TI - Autumn flu clinics are worth all the hassle. PMID- 16312074 TI - Younger care. PMID- 16312075 TI - A winning idea for nutrition. PMID- 16312076 TI - Advances and challenges in the therapeutic use of stem cells. AB - In spite of the monumental challenges, supporters of stem cell research remain convinced of its benefits to future generations. This article outlines what stem cells are and discusses their current therapeutic applications, current research, new potential uses, and the scientific and ethical challenges that these present. PMID- 16312077 TI - Administration of oral medication. PMID- 16312078 TI - Symptoms and management options in cardiomyopathy. AB - Cardiomyopathy is the most common cause of sudden cardiac death in young people, including trained athletes (Maron et al, 1996). Often the tragedy of sudden death is the first indication of its presence within a family. This article describes the four types of cardiomyopathy, diagnosis, treatment and management options, and discusses ways in which the new National Service Framework for Coronary Heart Disease (Department of Health, 2005) will help in planning and implementing services. PMID- 16312079 TI - A project to modernise the role of the ward manager. AB - Fractured neck of femur accounted for more than 1.7 million bed days in 2003 (Department of Health, 2003). This article discusses how the role of the ward manager on an older persons' trauma ward was modernised to provide a specialist service for outlying patients while maintaining ward management responsibility. This project is well established and has been successfully applied to coronary care. PMID- 16312080 TI - Intrauterine devices. PMID- 16312081 TI - Experiences and perceptions of music therapy in critical illness. AB - AIM: To gain insight into patients' and nurses' perception of the benefits of music therapy during critical illness. METHOD: The patients received music therapy on two prescribed occasions for 30 minutes each day. Unstructured interviews were conducted with five patients and four nurses and analysed according to Burnard's framework (1991). RESULTS: Patients can and do recall listening to music, even when they have been sedated and ventilated. They related to the music that they were listening to and some experienced imagery concerning the music. The nurses found it useful as an aid to wean patients off the ventilator, helping to keep them calm and focused. CONCLUSION: The study concluded that music is useful not only for relaxing patients but also in some cases 'relocating' them from the critical care area to the place where they would normally be listening to the music. PMID- 16312082 TI - How to manage the staff duty roster. PMID- 16312083 TI - Helping to alleviate the family's burden. PMID- 16312084 TI - Quality of care and the reality of a patient's life. PMID- 16312085 TI - The effects of Islam and traditional practices on women's health and reproduction. AB - The aim of this study was to investigate the effects of Islam as a religion and culture on Turkish women's health. The study included 138 household members residing in the territory of three primary health care centers in Turkey: Guzelbahce, Fahrettin Altay and Esentepe. Data were collected by means of a questionnaire prepared by a multidisciplinary team that included specialists from the departments of public health, psychiatric nursing and sociology. We found that the women's health behavior changed from traditional to rational as education levels increased, and that religious and traditional attitudes and behaviors were predominant in the countryside, especially practices related to pregnancy, delivery, the postpartum period, induced abortion and family planning. One of the most important prerequisites for the improvement of women's health is that nurses should know the religious practices and culture of the society for which they provide care, so that their efforts to protect and improve women's health will be effective. PMID- 16312086 TI - One size doesn't fit all: ethics of international nurse recruitment from the conceptual framework of stakeholder interests. AB - This theoretical study examines the ethics of international nurse recruitment from the conceptual framework of stakeholder interests. It argues that there are stakeholders at individual, institutional, national and international levels, with overlapping but, more often, different or even conflicting interests. Depending on the interests of given stakeholders, different conclusions regarding the ethics of international nurse recruitment may be reached. There is no right or wrong with these varying ethical positions because they reflect different beliefs and philosophies that are not amenable to value judgment. To illustrate and support this line of argument, this article analyzes the underpinnings of two ethical standards published by the International Council of Nurses and the UK Department of Health. In addition, a case study on China augments the argument by demonstrating limitations of the one-size-fits-all approach to the issue. The most important question in understanding and evaluating the ethical standards of international nurse recruitment is to know whose interests they are designed to represent and protect. PMID- 16312087 TI - Moral competence in nursing practice. AB - This article presents the derivation of moral competence in nursing practice by identifying its attributes founded on Thai culture. In this process moral competence is formed and based on the Thai nursing value system, including personal, social and professional values. It is then defined and its three dimensions (moral perception, judgment and behavior) are also identified. Additionally, eight attributes as indicators of moral competence are identified and selected from three basic values. The eight attributes are loving kindness, compassion, sympathetic joy, equanimity, responsibility, discipline, honesty, and respect for human values, dignity and rights. All attributes are discussed by addressing the three moral dimensions in order to present how to deal with ethical issues in nursing practice. As a summary, a model of moral competence is presented to demonstrate moral competence in nursing practice in Thailand. PMID- 16312088 TI - Sensitivity to ethical issues confronted by Korean hospital staff nurses. AB - This descriptive study was undertaken to identify the degree of ethical sensitivity of staff nurses and to analyze the differences in ethical sensitivity in terms of both general and ethics-related characteristics. Participants were 236 staff nurses working in general hospitals in Korea. Ethical sensitivity was measured by means of an instrument developed by the researchers. The results showed that the mean score for the degree of ethical sensitivity was 0.71 out of a possible maximum score of 1 (range 0.30 to 0.97). For general characteristics, there was a significant difference in ethical sensitivity according to age (F (df 2233) = 3.99, P = 0.02). For characteristics related to ethics, there was a significant difference in ethical sensitivity according to attitude towards the nursing profession (F (df 4231) = 2.94, P = 0.03). It is therefore recommended that a training program reflecting these variables be developed to enhance staff nurses' ethical sensitivity. PMID- 16312089 TI - Ways of relating during childbirth: an ethical responsibility and challenge for midwives. AB - The way in which midwives relate to expectant parents during the process of childbirth greatly influences the parents' childbirth experiences for a long time. We believe that examining and describing ways of relating in naturally occurring interactions during childbirth should be considered as an ethical responsibility. This has been highlighted in relation to parents' experiences and in the light of the relational ethics of Logstrup. Four couples' and nine midwives' ways of relating were documented by 27 hours of observation, including 14.5 hours of video-recorded sessions. A qualitative content analysis was conducted. The midwives strongly influenced the different ways of relating and three aspects of professional competence were disclosed. The results can contribute to reflections about current praxis as an ethical demand for midwives. PMID- 16312090 TI - Practising virtue: a challenge to the view that a virtue centred approach to ethics lacks practical content. AB - A virtue centred approach to ethics has been criticized for being vague owing to the nature of its central concept, the paradigm person. From the perspective of the practitioner the most damaging charge is that virtue ethics fails to be action guiding and, in addition to this, it does not offer any means of act appraisal. These criticisms leave virtue ethics in a weak position vis-a-vis traditional approaches to ethics. The criticism is, however, challenged by Hursthouse in her analysis of the accounts of right action offered by deontology, utilitarianism and virtue ethics. It is possible to defend the action guiding nature of virtue ethics: there are virtue rules and exemplars to guide action. Insights from Aristotle's practical approach to ethics are considered alongside Hursthouse's analysis and it is suggested that virtue ethics is also capable of facilitating action appraisal. It is at the same time acknowledged that approaches to virtue ethics vary widely and that the challenges offered here would be rejected by those who embrace a radical replacement virtue approach. PMID- 16312091 TI - Is emotional labour ethical? PMID- 16312092 TI - Exploiting the dead? The use of human cadavers for research, teaching and spare body parts: a multidisciplinary enquiry. PMID- 16312093 TI - The orchestral 'A'...nursing dissonance in the Middle East. PMID- 16312094 TI - Amnesty International urges a stronger human rights role for nurses and midwives. PMID- 16312095 TI - A laminar cortical model of stereopsis and 3D surface perception: closure and da Vinci stereopsis. AB - A laminar cortical model of stereopsis and 3D surface perception is developed and simulated. The model describes how monocular and binocular oriented filtering interact with later stages of 3D boundary formation and surface filling-in in the LGN and cortical areas V1, V2, and V4. It proposes how interactions between layers 4, 3B, and 2/3 in V1 and V2 contribute to stereopsis, and how binocular and monocular information combine to form 3D boundary and surface representations. The model includes two main new developments: (1) It clarifies how surface-to-boundary feedback from V2 thin stripes to pale stripes helps to explain data about stereopsis. This feedback has previously been used to explain data about 3D figure-ground perception. (2) It proposes that the binocular false match problem is subsumed under the Gestalt grouping problem. In particular, the disparity filter, which helps to solve the correspondence problem by eliminating false matches, is realized using inhibitory interneurons as part of the perceptual grouping process by horizontal connections in layer 2/3 of cortical area V2. The enhanced model explains all the psychophysical data previously simulated by Grossberg and Howe (2003), such as contrast variations of dichoptic masking and the correspondence problem, the effect of interocular contrast differences on stereoacuity, Panum's limiting case, the Venetian blind illusion, stereopsis with polarity-reversed stereograms, and da Vinci stereopsis. It also explains psychophysical data about perceptual closure and variations of da Vinci stereopsis that previous models cannot yet explain. PMID- 16312096 TI - Differential roles of distracters in reflexive and memory-based localization. AB - We investigated the effects of spatial and temporal factors on manual localization of a visual target by measuring accuracy, precision, and bias. Spatial factors included manipulation of display as with or without distracters, with invariant or variant distracters, and with near or far distracters, respectively, in Experiments 1, 2, and 3. The target and distracters were of 1degrees dots differing only by luminance parameter; they were presented concurrently for 150 or 1000 ms while observers had to memorize the target location maintaining a fixed gaze. The observers' task was to reproduce the location of the target with a mouse cursor available 150 ms following stimuli offset. Results from all experiments showed that localization performance for a briefly exposed target was as accurate and precise as that for a long exposed target. Moreover, manipulation of spatial factors had no systematic effects on accuracy and precision except that near distracters yielded higher precision. Interestingly, localization performance was unbiased in 150 ms condition when there were distracters in the display, while being biased towards the fovea in 1000 ms condition regardless of their presence or absence. These results suggest a temporal dynamics in dominance-suppression between egocentric and exocentric cues in the construction of memory for location. PMID- 16312097 TI - The changes in the tensile properties of tendons after freeze storage in saline solution. AB - The objective of this study is to evaluate the influence of saline solution (0.9 per cent NaCl) on the tensile properties of freeze-stored tendons. Firstly, 170 pieces of chicken flexor digitorum profundus tendons were retrieved and wrapped in saline-soaked gauze before they were stored at -40degreesC. Then specimens were tensile tested at various time points over 360 days, scanning electron microscopy (SEM) was performed on fresh specimens, and specimens were freeze stored for 233 days to investigate microstructure change after freeze storage. The mean values of strain ultimate tensile strength (UTS) did not deviate significantly (analysis of variance; p = 0.249) following freeze storage while the UTS and elastic modulus increased gradually with the duration of freeze storage and the growth became significant (p < 0.01) for durations longer than 70 and 40 days respectively. The SEM study showed that the collagen fibre density of specimens stored for 233 days decreased because of porosity growth. These findings suggested that the saline increased the tensile strength and modulus of the collagen. PMID- 16312098 TI - Three-dimensional measurement of intervertebral kinematics in vitro using optical motion analysis. AB - Measurement of the stiffness of spinal motion segments is widely used for evaluating the stability of spinal implant constructs. A three-dimensional motion analysis technique has been developed that allows accurate measurement of the relative movement of the vertebral bodies about a well-defined anatomical axis system. The position of marker clusters on each vertebra is tracked using digital infrared cameras (Qualisys AB, Gothenburg). Landmarks are identified using a marked pointer, and an anatomical coordinate system is defined for each vertebra. The transformation relating the upper and lower vertebrae is calculated, using the joint coordinate system approach of Grood and Suntay to find the rotations and translations in each anatomical plane. The stiffness of vertebrectomy constructs was investigated using a Synex vertebral body replacement and an anterior rod with one or two screws in each vertebral body, with or without damage to the posterior longitudinal ligament (PLL). A moment of 2 N m was applied about each anatomical axis, and the range of motion about each axis was calculated. The range of motion in flexion-extension and lateral bending was significantly greater with only one screw. When the PLL was cut, there was no significant increase in the range of motion. PMID- 16312099 TI - Precision and accuracy in ceramic-on-ceramic wear analyses: influence of simulator test duration. AB - In this, the first report of precision and accuracy in simulator studies, ceramic ceramic implants with ultra-low wear trends represented a relevant wear model. The effect of test durations was examined in a standard simulator test mode on the quality of the linear regression trends, the average wear estimates, and the amount of noise in the data. Three sets of diametral tolerances were compared in 28 mm diameter alumina implants. The authors' hypothesis was that wear data would be significantly improved with increased test durations. The average wear rates varied little with test duration, the biggest change amounting to only 30 and 15 per cent decreases in the wear estimate by 10 and 14 million cycles respectively. The most satisfactory improvement in the study was the decrease in variance (noise) with increasing duration, +/- 200 per cent at 5 million cycles reduced to +/- 55 per cent at 14 million cycles. The quality of the linear regression coefficients improved 150 per cent by 10 million cycles and 250 per cent by 14 million cycles. Overall the ceramic implants with highest diametral tolerances showed the least wear (15 per cent less, but not statistically significant). However, given such low wear rates for alumina liners, it was unlikely that any differences owing to diametral tolerances would be clinically significant in the typical patient. PMID- 16312100 TI - An in-vitro investigation into the cement pressurization achieved during insertion of four different femoral stems. AB - Adequate cement pressurization during stem insertion improves the interdigitation of cement into bone. This increases the strength of the cement-bone interface, thus contributing to the reduction of the incidence of aseptic loosening, the commonest cause of revision surgery. This in-vitro study compared the cement pressurization achieved during insertion of four different stems of equivalent sizes: the Elite Plus (DePuy, UK), C-Stem (DePuy, UK), Exeter (Stryker, USA), and CPS-Plus (Plus Orthopedics, Switzerland). The maximum pressures attained at the time of stem insertion were recorded at proximal, mid and distal stem levels. The Elite Plus generated significantly higher distal pressures than the other stems. The CPS-Plus generated significantly greater proximal cement pressures than the Elite Plus, C-Stem, and Exeter prostheses. The triple taper of the C-Stem increased the cement pressurization medial to the stem. The stem shape and the presence or absence of a proximal stem centralizer affect cement pressurization. The presence of a proximal stem centralizer, a large stem volume, and a lateral medial taper are all factors associated with increased cement pressurization during stem insertion. PMID- 16312101 TI - A three-dimensional dynamic finite element model of the prosthetic knee joint: simulation of joint laxity and kinematics. AB - For testing purposes of prostheses at a preclinical stage, it is very valuable to have a generic modelling tool, which can be used to optimize implant features and to avoid poor designs being launched on to the market. The modelling tool should be fast, efficient, and multi-purpose in nature; a finite element model is well suited to the purpose. The question posed in this study was whether it was possible to develop a mathematically fast and stable dynamic finite element model of a knee joint after total knee arthroplasty that would predict data comparable with published data in terms of (a) laxities and ligament behaviour, and (b) joint kinematics. The soft tissue structures were modelled using a relatively simple, but very stable, composite model consisting of a band reinforced with fibres. Ligament recruitment and balancing was tested with laxity simulations. The tibial and patellar kinematics were simulated during flexion-extension. An implicit mathematical formulation was used. Joint kinematics, joint laxities, and ligament recruitment patterns were predicted realistically. The kinematics were very reproducible and stable during consecutive flexion-extension cycles. Hence, the model is suitable for the evaluation of prosthesis design, prosthesis alignment, ligament behaviour, and surgical parameters with respect to the biomechanical behaviour of the knee. PMID- 16312102 TI - Effect of interface conditions between ultrahigh molecular weight polyethylene and polymethyl methacrylate bone cement on the mechanical behaviour of total shoulder arthroplasty. AB - The aim of this study was to investigate the effect of the interface condition between polymethyl methacrylate (PMMA) bone cement and the ultrahigh molecular weight (UHMWPE) glenoid component on cement stresses and glenoid component tilting in a finite element (FE) model. The background of this research is that most FE models assume bonding between the PMMA bone cement and the UHMWPE component, although it is very doubtful that this bonding is present. An FE model of a cemented glenoid component was developed and a joint compression force and subluxation force of 725 and 350 N respectively were applied. The maximal principal stresses in the cement layer ranged between 21.30 and 32.18 MPa. Glenoid component tilting ranged between 0.943 degrees and 0.513 degrees. It was found that the interface condition has a large effect on the maximal principal stresses and glenoid component tilting. Whether adhesion between the UHMWPE component and PMMA bone cement occurs is unknown beforehand and, as a result, design validation using the FE technique should be carried out both by using contact elements in combination with a coefficient of friction as well as by a full bonding at this interface. PMID- 16312103 TI - A 12-station anatomic hip joint simulator. AB - A novel 12-station hip joint simulator with an anatomic position of the prosthesis was designed and built. The motion of the simulator consists of flexion-extension and abduction-adduction. The load is of the double-peak type. The validation test was done with three similar 28 mm CoCr-polyethylene joints in diluted calf serum lubricant for 3.3 x 10(6) cycles. The bearing surfaces of the polyethylene cups were burnished, the CoCr heads were undamaged, the wear particles were in the 0.1-1 microm size range, and the mean wear factor of the polyethylene cups was 5.7 x 10(-7) mm(3)/N m. These essential observations were in good agreement with clinical findings. In addition, three similar 50 mm CoCr/CoCr joints, representing the contemporary large-diameter metal-on-metal articulation were tested. The wear of the CoCr/CoCr joints was calculated from the Co and Cr concentrations of the used lubricant quantified with atomic absorption spectroscopy. The bearing surfaces of the CoCr/CoCr joints showed mild criss-cross scratching only. The average wear factor of polyethylene cups was 275 times that of the CoCr/CoCr joints. The tribological behaviour of the large-dia. CoCr/CoCr appeared to be dominated by fluid film lubrication, as indicated by very low frictional heating and wear, making it tribologically superior to the conventional CoCr/polyethylene, and therefore very interesting clinically. In conclusion, the simulator proved to be a valid, reliable, practical, economical, and easy-to-operate tool for wear studies of various hip replacement designs. PMID- 16312104 TI - A circumferentially flanged tibial tray minimizes bone-tray shear micromotion. AB - Aseptic loosening of the tibial component is the major complication of total knee arthroplasty. There is an association between early excessive shear micromotion between the bone and the tray of the tibial component and late aseptic loosening. Using non-linear finite element analysis, whether a tibial tray with a circumferentially flanged rim and a mating cut in the proximal tibia could minimize bone-tray shear micromotion was considered. Fifteen competing tray designs with various degrees of flange curvature were assessed with the aim of minimizing bone-tray shear micromotion. A trade-off was found between reducing micromotion and increasing peripheral cancellous bone stresses. It was found that, within the limitations of the study, there was a theoretical design that could virtually eliminate micromotion due to axial loads, with minimal bone removal and without the use of screws or pegs. PMID- 16312105 TI - Fluid composition impacts standardized testing protocols in ultrahigh molecular weight polyethylene knee wear testing. AB - Wear of total knee replacements is determined gravimetrically in simulator studies. A mix of bovine serum, distilled water, and additives is intended to replicate the lubrication conditions in vivo. Weight gain due to fluid absorption during testing is corrected using a load soak station. In this study, three sets of ultrahigh molecular weight polyethylene tibial plateau were tested against highly polished titanium condyles. Test 1 was performed in two different institutions on the same simulator according to the standard ISO 14243-1, using two testing lubricants. Test 2 and test 3 repeated both previous test sections. The wear and load soak rates changed significantly with the lubricant. The wear rate decreased from 16.9 to 7.9 mg weight loss per million cycles when switching from fluid A to fluid B. The weight gain of the load soak specimen submersed in fluid A was 6.1 mg after 5 x 10(6) cycles, compared with 31.6 mg for the implant in fluid B after the same time period. Both lubricants were mixed in accordance with ISO 14243 (Implants for surgery - wear of total knee-joint prostheses), suggesting that calf serum should be diluted to 25 +/- 2 per cent with deionized water and a protein mass concentration of not less than 17 g/l. The main differences were the type and amount of additives that chemically stabilize the lubricant throughout the test. The results suggest that wear rates can only be compared if exactly the same testing conditions are applied. An agreement on detailed lubricant specifications is desirable. PMID- 16312106 TI - Mass transport disturbances in the distal graft/artery junction of a peripheral bypass graft. AB - Intimal hyperplasia (IH) development is a primary cause of failure of reconstructive bypass surgery. While the exact mechanism by which IH initiates and proliferates has yet to be fully elucidated, it is clear that the abnormal haemodynamics present in the downstream graft/artery junction are intrinsic in its development. Mass transport disturbances owing to abnormal haemodynamics have been associated with atherogenesis and it is for this reason that an investigation into transport of platelet-derived growth factor (PDGF), a known promoter of the intimal hyperplastic response, at the downstream graft/artery junction was carried out. A steady flow analysis in a three-dimensional, idealized, downstream graft/artery junction was carried out using commercial computational fluid dynamics software. It was found that there is a two-and-half fold increase in the transport of PDGF to the artery wall at the bed of the junction when compared with an idealized, healthy artery. The presence of secondary flows in the downstream arterial section also leads to large disturbances in mass transport. It was concluded that PDGF transport in the downstream graft/artery junction tends to be highly disturbed and that there may be a role of this disturbance in the initiation and subsequent development of distal anastomotic intimal hyperplasia. PMID- 16312107 TI - A first approach to in vitro simulation of vascular calcification by the controlled crystallization of poorly crystalline calcium phosphates onto porous cholesterol. AB - A first approach to in vitro simulation of vascular calcification was elaborated. Vascular calcification was simulated by a slow crystallization of a non stoichiometric poorly crystallized carbonateapatite from Kokubo's revised simulated body fluid (rSBF) on the surface of a porous pellet made of pure cholesterol. To achieve this, the crystallization experiments were performed under strictly controlled conditions (similar to physiological ones) provided by a constant-composition double-diffusion (CCDD) device. To obtain an even closer match to in vivo conditions, rSBF was enriched by the addition of glucose and bovine serum albumin (BSA) in physiological amounts. Precipitation took place on the surface of cholesterol and the precipitates consisted of poorly crystalline non-stoichiometric, sodium- and magnesium-containing carbonateapatite. PMID- 16312108 TI - University students' knowledge of alcoholic drinks and their perception of alcohol-related harm. AB - A total of 371 university students were asked to estimate the amount of alcohol contained in a standard drink and to estimate the number of standard drinks contained in popular alcoholic beverages. In addition, students completed questionnaires assessing their perception of short and long term harm related to the consumption of beer, wine, spirits and pre-mixed alcopops. Results revealed that students were generally inaccurate in their estimate of alcoholic content of beverages, and national guidelines for low risk drinking. Students were also found to hold different perceptions regarding how harmful different alcoholic beverages were. While both male and female students considered spirits to be more harmful than beer, wine and pre-mixed drinks, males also believed beer to be more harmful than wine when considering the short term consequences. The pattern of beliefs reported by this sample suggest a high-risk population who are not aware of the risks they are exposing themselves to through their drinking behavior. PMID- 16312109 TI - Alcohol expectancies in relation to personality and aggression among juvenile delinquents in Northern Russia. AB - The relationships between alcohol expectancies, level of alcohol use, alcohol related problems, aggression, and personality factors in 198 Russian male juvenile delinquents were assessed. A clustering procedure was used in order to establish main patterns of alcohol expectancies, yielding three major clusters. Level of alcohol use, alcohol-related problems, aggression, and personality factors were compared across the identified clusters. It was established that juvenile delinquents with a high level of positive alcohol expectancies and aggression represented a risk-group for higher involvement in drinking behavior as well as problem drinking, which in turn are related to specific personality traits. Implications of these findings for alcohol prevention among the youth are discussed. PMID- 16312110 TI - Adolescents' beliefs about marijuana use: a comparison of regular users, past users and never/occasional users. AB - A questionnaire investigating adolescents' opinions and experiences regarding marijuana use was administered to 163 adolescents and young adults (96 boys and 67 girls) aged 13 to 20 (mean age = 16.8, s.d. = 1.5). Items referred to marijuana and other substances' dangerousness, representations regarding the positive and negative consequences of marijuana use. Responses were compared according to marijuana use status (classified into never/occasional use, current regular use and past regular use). Results show that adolescents' opinions differ according to their experience with marijuana use. Current regular users evaluate marijuana as less dangerous, but alcohol and heroin as more dangerous in comparison with never/occasional and past users. Current and past users are more likely to define marijuana as a medical drug and a plant used in agriculture, and less likely to define it as an illegal drug. Current and past users evaluate marijuana use as a way to cope with stress, to relax to a greater extent than do never/occasional users do. The latter attribute more negative consequences to marijuana use such as diminished driving ability and school performance and a pathway to hard drugs. PMID- 16312111 TI - Drinking motives, alcohol expectancies, self-efficacy, and drinking patterns. AB - The current study focused on the associations between drinking motives, alcohol expectancies, self-efficacy, and drinking behavior in a representative sample of 553 Dutch adolescents and adults. Data were gathered by means of self-report questionnaires and a 14-days drinking diary. A model was postulated in which negative expectancies and self-efficacy were directly associated with drinking, and in which drinking motives mediated the associations between positive expectancies, and drinking. The findings of multivariate analyses showed that drinking motives were related to general indicators of drinking and to drinking levels in specific situations. Furthermore, self-efficacy was moderately related to all drinking variables. Negative expectancies were related to general drinking variables but hardly to drinking in specific situations. Positive expectancies were hardly related to drinking in multivariate analyses and therefore mediation models could not be tested. No systematic moderator effects were apparent for age and gender on the associations between drinking motives, alcohol expectancies, self-efficacy, and drinking. PMID- 16312112 TI - Student nurse gives antibiotic i.v.--death results. Dennis v. Specialty Select Hospital-Flint, 2005 WL 2402454 N.W.2d -MI. PMID- 16312113 TI - Could this nurse midwife deliver for you or yours? O'Connor v. Washington DOH, Nursing Care Q.A. Com., 2005 WL 2338685, P.2d -WA. PMID- 16312114 TI - TX: nurses fail to provide bedpan or assistance: was their negligence cause of fall and fracture? Torres v. Memorial Hospital System, 2005 WL 2615312 S.W.3d TX. PMID- 16312115 TI - Nurse temporarily in ICU increases dosage without authorization. Woodward v. Research Medical Center, 2005 WL 2007878 S.W.3d -MO. PMID- 16312116 TI - Special care baby charter: an exercise in the obvious? PMID- 16312117 TI - Babies at risk from mothers' diabetes. PMID- 16312118 TI - Hospital Trust supports the RCM 'Campaign for normal birth'. PMID- 16312119 TI - Health libraries week 2005. PMID- 16312120 TI - Looking back to go forward: Tair Afon--the first birth centre in Wales. PMID- 16312121 TI - Retaining valued midwives. AB - In a Delphi study of 42 experienced midwives at an NHS Trust in south-east England, it was established that leadership issues were at the centre of what encouraged practitioners to continue in post. Central to midwives' interest was the equitable management of off-duty/leave arrangements, opportunities to influence practice and protocol decision-making, feeling stimulated, valued and challenged as a practitioner, receiving professional development support and working with senior midwives who proved effective role models. Beyond reward opportunities associated with Agenda for Change or other initiatives, it is argued that midwifery managers have a key opportunity to influence the employment experience of midwives and enhance retention of staff by actively engaging in the above activities. PMID- 16312122 TI - Is there such a thing as a free lunch? PMID- 16312123 TI - Members' chance to speak out: why not propose a motion for annual conference? PMID- 16312124 TI - Are you normal? Implementing normality in the maternity unit. PMID- 16312125 TI - International Confederation of Midwives congress in Brisbane. PMID- 16312126 TI - Criticism over bottle-feeding research. PMID- 16312127 TI - Criticism over bottle-feeding research. PMID- 16312128 TI - Cytokines: the magic is still there. PMID- 16312129 TI - Mitochondria-dependent apoptosis in T-cell homeostasis. AB - T-cell homeostasis is maintained by balancing the proliferation and destruction of lymphocytes at multiple steps during the life of an individual. Regulated mitochondria-dependent apoptosis is essential for both the development and the subsequent maintenance of the immune system, in that it keeps the total number of lymphocytes constant. Firstly, during thymic development, sequential stages of T cell maturation require strict control of T-cell selection, and secondly, apoptosis is essential in controlling the massive expansion of antigen-specific T cells after their activation. Failure in each of these steps can lead to pathologies, while drugs that target apoptosis could have therapeutic benefit. PMID- 16312130 TI - Adhesion molecules in human dendritic cells. AB - Dendritic cells are bone marrow-derived professional antigen-presenting cells that link innate and adaptive immunity, and they are essentially involved in the initiation of primary immune responses and in the establishment of peripheral tolerance. The existence of distinct functional states and subsets of dendritic cells is critical for the generation of pathogen-specific immune responses without the risk of autoimmunity or chronic inflammation. To fulfil their effector tasks in tissues and lymph nodes, dendritic cells must engage in multiple adhesive and migratory events. The molecular dissection of these adhesive interactions may provide new potential therapeutic targets to modulate immune responses and to improve current dendritic cell-based therapeutic cancer vaccines. PMID- 16312131 TI - Lipoxin analogs: novel anti-inflammatory mediators. AB - Most pharmacological anti-inflammatory agents are designed to inhibit the enzymes or receptors that are directly involved in mediating inflammation. In contrast, lipoxin analogs utilize an entirely different anti-inflammatory paradigm, namely that of activating the endogenous anti-inflammatory signaling pathways by which inflammation is normally resolved. Specifically, synthetic stable analogs of the endogenous anti-inflammatory eicosanoid lipoxin A4 (LXA4) have been shown to mimic the anti-inflammatory actions of LXA4 in a variety of inflammation models and display desirable pharmacological properties, especially their high oral bioavailability. Lipoxin analogs could be potentially useful for a wide array of disease states that have an inflammatory component, and look particularly promising for treatment of inflammatory bowel disease. PMID- 16312132 TI - Role of vasoactive intestinal peptide in inflammation and autoimmunity. AB - Vasoactive intestinal peptide (VIP), a peptide produced by immune cells, exerts a wide spectrum of immunological functions that control the homeostasis of the immune system. In the last decade, VIP has been clearly identified as a potent anti-inflammatory factor, both in innate and adaptive immunity. In innate immunity, this peptide inhibits the production of inflammatory cytokines and chemokines from macrophages, microglia and dendritic cells. In addition, VIP reduces the expression of co-stimulatory molecules on antigen-presenting cells, and therefore reduces stimulation of antigen-specific CD4 T-cells. In terms of adaptive immunity, VIP promotes T-helper (Th)2-type responses, and reduces inflammatory Th1-type responses. Several of the molecular mechanisms involved in the inhibition of cytokine and chemokine expression, and in the preferential development and/or survival of Th2 effectors are known. Therefore, VIP and its analogs have been proposed as promising alternative candidates to existing therapies for the treatment of acute and chronic inflammatory and autoimmune diseases. The aim of this review is to update knowledge of the cellular and molecular events that are relevant to VIP function in the immune system. The central functions that VIP plays in cellular processes is being recognized and attention is being focused on this important peptide with regard to exciting new candidates for therapeutic intervention and drug development. PMID- 16312133 TI - The involvement of VEGF in endothelial permeability: a target for anti inflammatory therapy. AB - Many inflammatory mediators are known to contribute to increased vascular permeability during various phases of inflammation. Among these mediators, vascular endothelial growth factor (VEGF), also known as vascular permeability factor, plays an important role in vascular hyper-permeability, and several studies have demonstrated that modulation of VEGF function might contribute to a successful therapeutic approach to the treatment of inflammatory diseases. VEGF receptor monoclonal antibodies, glucocorticoids, leukotriene modifiers, cyclooxygenase-2 inhibitors and non-steroidal anti-inflammatory drugs have been studied for many years as VEGF-blocking drugs in inflammatory diseases in vitro and/or in vivo. The molecular and biological understanding of VEGF and preclinical or clinical trials of VEGF-blocking therapies raise hope that they may be useful as adjuncts to existing anti-inflammatory approaches in the regulation of inflammatory diseases. PMID- 16312134 TI - Tick histamine-binding proteins and related lipocalins: potential as therapeutic agents. AB - Tick histamine-binding proteins bind histamine with high affinity and specificity. This is attained by a novel binding mechanism, whereby histamine is sequestered within a binding cavity of the lipocalin fold. The histamine binding proteins and related protein family members are currently under investigation as potential therapeutic agents for the treatment of various diseases, including conjunctivitis, allergic rhinitis, carcinoid syndrome and rheumatoid arthritis. While these proteins show great therapeutic potential, they are part of a diverse family of tick lipocalin proteins, some of which have been implicated in tick host rejection and host pathogenesis. As such, the therapeutic mining of tick lipocalins should be considered within the framework of the rest of the family. PMID- 16312135 TI - Selective phosphodiesterase 4 inhibitors in the treatment of allergy and inflammation. AB - Phosphodiesterase (PDE)4 inhibitors are a novel class of drugs in development for the treatment of inflammatory airways diseases, including asthma, allergic rhinitis and chronic obstructive pulmonary disease. PDE4 inhibitors are potent anti-inflammatory agents both in vitro and in vivo, but few have successfully proceeded to phase II and III clinical trials, as a result of insufficient clinical efficacy and unacceptable side effects, including nausea and emesis, which have hampered their progression. A greater understanding of the molecular biology of PDE4 has led to the development of efficacious compounds with fewer side effects. This review focuses on how selective PDE4 inhibitors can advance the treatment of airways diseases and deal with the challenges that lie ahead. PMID- 16312136 TI - Potential immunosuppressive therapeutics to target small vessel vasculitis of the kidney. AB - Small vessel vasculitis of the kidney is an inflammatory disorder that results in glomerulonephritis. Current treatment regimes have proved effective, with patient survival rates of 80% at five years. Of the 20% that die, a significant number of deaths are due to the side effects of therapies, and immunosuppression. A more tailored approach is required that targets the cells involved rather than generally restraining the immune system. This can only be achieved by increasing our knowledge of disease pathogenesis. PMID- 16312137 TI - AWD-12-281 (inhaled) (elbion/GlaxoSmithKline). AB - elbion (formerly ASTA Medica) and GlaxoSmithKline are developing an inhaled formulation of AWD-12-281 for the potential treatment of chronic obstructive pulmonary disease (COPD). By May 2005, phase II trials of this 5-hydroxyindole PDE4 inhibitor for COPD were ongoing. PMID- 16312138 TI - Retisert (Bausch & Lomb/Control Delivery Systems). AB - Retisert (Envision TD), a sustained release intraocular implant containing fluocinolone acetonide, has been developed and launched in the US by Bausch & Lomb and Control Delivery Systems for the treatment of chronic non-infectious uveitis affecting the posterior segment of the eye. PMID- 16312139 TI - Febuxostat (Teijin/Ipsen/TAP). AB - Febuxostat (TEI-6720; TMX-67), an oral non-purine, selective inhibitor of xanthine oxidase (NP-SIXO), is being developed by Teijin, with licensees Ipsen and TAP Holdings, for the potential treatment of gout. In February 2005, it was reported that launch in Japan was expected to be delayed for several years due to the need for additional clinical trials; in May 2005, an EU filing was expected by the end of 2005. PMID- 16312140 TI - Velafermin (CuraGen). AB - CuraGen is developing velafermin (CG-53135), human fibroblast growth factor-20 administered intravenously, for the potential treatment and prevention of oral mucositis (OM). By December 2004, phase II trials for the prevention of OM in cancer patients undergoing bone marrow transplant had been initiated, and in September 2005 CuraGen indicated that results from this study will be reported during the first quarter of 2006. PMID- 16312141 TI - Parkin expression in muscle from three patients with autosomal recessive Parkinson's disease carrying parkin mutation. AB - Absence of parkin has been shown to cause the downturned wing phenotype and severe disruption of myofibrils and mitochondrial abnormalities in parkin null mutant drosophila. The present report refers to studies on 3 patients with autosomal recessive Parkinson's disease with mild histopathological changes in muscle and a 'G' deletion at the exon9/intron9 junction or exon 3 deletion in the parkin gene. Using an antibody against a peptide corresponding to sequence number 305-323 of the human parkin protein it was demonstrated that parkin was expressed in skeletal muscle of these patients and that its distribution was similar to that in normal muscles. The mild nature of the histopathological changes, especially in the young patients, in the presence of parkin mutations may be due to residual E3-ubiquitin ligase activity of the mutant protein, to existence of muscle-specific splice-variants or to the relative functional insignificance of parkin in human muscle fibres. Further investigation of parkin expression in skeletal muscle is warranted. PMID- 16312142 TI - Long-term MRI and clinical follow-up of symptomatic and presymptomatic carriers of dysferlin gene mutations. AB - We report the results of a longitudinal study involving MRI and clinical follow up in nine siblings from four families with Miyoshi myopathy (MM). All individuals carried pathogenic dysferlin gene (DYSF) mutations with six of them suffering from symptomatic disease and three being presymptomatic. In presymptomatic subjects, MRI was sensitive to detect alterations in muscle tissue years before disease onset. The first MRI alteration to disclose was evidence for myoedema in dorsal compartment muscles of the legs followed by fatty degeneration. Moreover, MRI changes anticipated the topography of subsequent clinical muscle involvement and progressed from distal to proximal dorsal leg muscles. In symptomatic subjects, MRI changes reflected the pattern and severity of clinical muscle involvement. MRI evidence, however, suggests that muscle involvement is much more prominent in early disease stages than clinically seen. Clinical follow-up up to 8 years made evident that MM onset occurs at a mean age of 18.4 years. The most prominent initial deficit was impaired tiptoe gait due to muscle plantarflexor dysfunction followed by impaired dorsiflexor function. Dorsal compartments were predominantly affected not only in distal but also in proximal leg muscles, and a more rapid progression was noticed during the early phase of the disease. Our data suggest that MRI is a helpful diagnostic tool for an early diagnosis of MM and other distal myopathies since it provides sensitive and topographic information about initial and even preclinical muscle involvement. This is of particular relevance in Miyoshi myopathy because distinct CK elevation is present long before its clinical onset and often misdiagnosed as "idiopathic". PMID- 16312143 TI - Fourth round table conference in Monaco on 15 January 2005: regulation of muscle growth, a therapeutic issue for Duchenne muscular dystrophy? PMID- 16312144 TI - The challenges of process analytical technologies in chemical development. PMID- 16312145 TI - Developments in the design and synthesis of calpain inhibitors. AB - Calpains are Ca(2+)-dependent cysteine proteases that play an important role in cell differentiation and in apoptosis/necrosis. The overactivation of calpain is connected with a number of diseases, including cataracts and traumatic brain injury, making calpain an attractive drug target. The development of selective inhibitors of calpain has, however, proved difficult, due to a lack of detailed structural information on the protein. This difficulty has been somewhat alleviated with recent reports on the X-ray crystal structures of engineered calpains and improved biochemical characterization of the protein. This review describes properties and X-ray crystal structures of calpain, and the synthesis and binding affinities of novel calpain inhibitors. PMID- 16312146 TI - Applications of microwave-assisted organic synthesis on the multigram scale. AB - This review presents a summary of reactions performed using microwave irradiation on a multigram scale. Results are described in the context of existing equipment, and equipment currently in development, for the processing of multigram to kilogram quantities of materials including single- and multimode microwave reactors and batch, batch-flow and continuous-flow systems. Although limited in number, reports found in the literature to date suggest that with appropriate controls, reproducible scale-up of microwave reactions is feasible and requires minimal re-optimization of laboratory-scale reaction conditions. This feature, along with the dramatic reductions in reaction time generally observed for microwave reactions suggests that application of this technology in process research could be beneficial. PMID- 16312147 TI - Practical enantioselective process for a chiral phosphodiesterase-4 inhibitor. AB - L-869298 is a potent and selective phosphodiesterase-4 (PDE4) inhibitor, which is potentially useful in the treatment of asthma and chronic obstructive pulmonary disorder. A catalytic asymmetric synthesis that is suitable for the preparation of kilogram quantities of L-869298, and which does not require the use of chromatography, has been developed to support the on-going drug development program of L-869298 at Merck Research Laboratories. The catalytic asymmetric hydrogenation of an aromatic heteroaromatic ketone afforded the corresponding alcohol in almost perfect enantioselectivity. Activation of the alcohol via formation of the 4-toluenesulfonate, followed by an unprecedented displacement of the tosylate via the lithium enolate of ethyl-3-pyridyl acetate N-oxide, generated the chiral tetra-substituted ethane. The displacement reaction proceeded with inversion of configuration and without loss of optical purity. Deprotection of the displacement adduct followed by decarboxylation, afforded L 869298 in excellent overall yield. The methodology developed could be readily extended to the synthesis of several other chiral PDE4 inhibitors. PMID- 16312148 TI - Furans, thiophenes and related heterocycles in drug discovery. AB - The five-membered aromatic heterocycles containing oxygen and sulfur are key building blocks used in medicinal chemistry applications. Furans, thiophenes and related azole analogs constitute an important class of compounds that are readily available, stable and easily functionalized. In this review, the authors survey the recent literature and highlight the application of such compounds in the development of therapeutic agents. This review focuses on two major strategies involving these compounds. The first is the use of these rings as intact scaffolds within the final target compound, and the second is the use of these compounds as convenient and flexible synthons for non-aromatic structural moieties. PMID- 16312149 TI - Approaches to the synthesis of immunolides: selective immunomodulatory macrolides for cystic fibrosis. AB - The discovery of the clinical effectiveness of erythromycin and azithromycin in inflammatory airway diseases has inspired the discovery and development of macrolides with selective immunomodulatory activity. Erythromycin degradation continues to be a source of novel macrolides with a variety of selective biological activities. New technologies for drug discovery based in the emerging field of combinatorial biosynthesis provide the medicinal chemist with novel approaches toward the discovery of novel macrolides. Recent efforts to integrate synthetic organic medicinal chemistry with combinatorial biosynthesis have expanded the number of techniques available for macrolide synthesis. PMID- 16312150 TI - Biosynthetic engineering of natural products for lead optimization and development. AB - It is now possible to rapidly and rationally modify, at a genetic level, the machinery responsible for natural product biosynthesis. This provides the opportunity to design new structures and to optimize natural product lead compounds in a way that would be extremely difficult through synthetic chemistry means alone. The technology can also be used to overcome limitations of compound supply, which might otherwise preclude natural products from progressing into clinical trials. Described herein are some recent examples which highlight how biosynthetic engineering has been applied to drug discovery and development, and which attempt, in particular, to demonstrate how the technology functions most effectively when combined with synthetic organic and medicinal chemistry. PMID- 16312151 TI - Polymer-assisted solution-phase chemical library synthesis. AB - Polymer-assisted solution-phase (PASP) synthesis has become a prevalent method for the parallel synthesis of chemical libraries. PASP methodology enables the chemist to prepare libraries of compounds more quickly and efficiently than previously. PASP technology encompasses a number of purification techniques for intermediate and final product purification. This methodology continues to progress, providing the practitioner with a broad range of ingenious purification methods, enabling multi-step syntheses of complex compounds. This review covers significant new advances in the use of tagged-reagents/substrates, resin capture release techniques and PASP multi-step synthesis, some of which incorporate microwave heating and automation. PMID- 16312152 TI - Recent advances in heterocycle generation using the efficient Ugi multiple component condensation reaction. AB - The current trend of rising research spending and falling numbers of novel chemical entities continues to drive efforts aimed at increasing efficiency in the drug discovery process. Strategic issues, such as assigning resources to poorly validated targets have been implicated in the declining productivity of recent years. Tactical approaches employed to improve this situation include attempts to speed the discovery process toward decision points in a timely manner. Accelerating the optimization of high-throughput screening hits is a goal in streamlining the discovery process, and the use of multiple-component condensation (MCC) reactions have proved useful toward this end. MCC reactions are powerful and efficient tools for the generation of diverse compound sets. Collections of compounds can be synthesized with all of the required diversity elements included in a single synthetic step. One of the most widely investigated MCC reactions is the Ugi four-component condensation. This review highlights disclosures of the Ugi reaction published over the past two years (2003 to 2005) in three areas: (i) Ugi reaction in conjunction with post-condensation cyclization; (ii) bifunctional condensations leading to heterocyclic cores; and (iii) general findings relating to linear products or interesting improvements in the basic Ugi reaction. PMID- 16312153 TI - Building process knowledge using inline spectroscopy, reaction calorimetry and reaction modeling--the integrated approach. AB - For over two decades, reaction engineering tools and techniques such as reaction calorimetry, inline spectroscopy and, to a more limited extent, reaction modeling, have been employed within the pharmaceutical industry to ensure safe and robust scale-up of organic reactions. Although each of these techniques has had a significant impact on the landscape of process development, an effective integrated approach is now being realized that combines calorimetry and spectroscopy with predictive modeling tools. This paper reviews some recent advances in the use of these reaction engineering tools in process development within the pharmaceutical industry and discusses their potential impact on the effective application of the integrated approach. PMID- 16312154 TI - Man, machine and visual imagery in strategic synthesis planning: computer perceived precursors for drug candidates. AB - This article provides a brief overview of methods for computer-perceived generation of precursors and starting materials to organic molecules. Emphasis is placed on software with validated performance in two main areas dealing with retrosynthetic analysis and heuristic perception of structural, functional and stereochemical similarities between target molecules and potential precursors. Applications to strategic syntheses of drug candidates are demonstrated. PMID- 16312155 TI - Asthma care in the new millennium. PMID- 16312156 TI - Rethinking asthma from business to bedside. AB - OBJECTIVE: To explore the possible application of concepts, methods, and tools validated in the worlds of statistics, business, engineering, and system dynamics to improve insight and outcomes in asthma care. DATA SOURCES: Peer-reviewed articles, books, and other publications on the topic of asthma care, dating back to 1931, were retrieved using MEDLINE searches and other external sources. Searches were restricted to include only studies with human subjects and the English language as descriptors, and the terms used included the following: system dynamics, statistical process control charts, continual improvement, mental models, personal mastery, systems thinking, team learning, and shared vision. Personal author observations and experiences were also included. STUDY SELECTION: Review and original research studies that reported on these concepts were evaluated. RESULTS: Results show that the concepts behind these models may be transferred to the system of asthma care. Indeed, case studies have shown that control charts can be used to detect signals of improved or declined function in asymptomatic patients, monitor medication adverse effects and the effects of adding a new medication, and evaluate the effectiveness of different medication delivery systems. CONCLUSIONS: These concepts and tools can provide immeasurable value to the process of rethinking and redesigning asthma care. They have the potential to help physicians and patients alike in designing and executing systems of care that are capable of not only delivering better outcomes but also inviting commitment to the long-term care many require. PMID- 16312157 TI - Asthma disease management: a critical analysis. AB - OBJECTIVE: To examine how effective a number of asthma disease management (DM) programs have been and whether they have been shown to improve patient outcomes cost-effectively. DATA SOURCES: Articles and meta-analyses from peer-reviewed journals or conference abstracts from 1991 to November 2003 were reviewed by searching MEDLINE and the external World Wide Web to provide examples of data. Terms used for searches included outcomes, intervention, specialist, self management, asthma, pediatric, and cost-effectiveness. STUDY SELECTION: Observational studies that reported data on DM programs, comparisons of general and specialty care, or asthma self-management outcomes were selected for review. RESULTS: Recently, several studies have demonstrated the short-term benefit of asthma DM programs. These studies found that educational intervention programs produce measurable clinical and economic benefits in the intervention group; however, only a small percentage of patients remain compliant. In addition, the benefit of specialty care in asthma has been shown in other studies. CONCLUSIONS: Although long-term studies are needed to evaluate DM-mediated clinical and cost benefits compared with specialty care, any program that helps improve patient compliance will result in clinical and economic benefits. PMID- 16312158 TI - Asthma in rural America. AB - OBJECTIVES: To examine current trends of asthma prevalence, diagnosis, and utilization of care in rural areas, increase awareness about asthma disease burden, and provide general recommendations for improving care in this setting. DATA SOURCES: MEDLINE searches were conducted for articles published from 1966 onward, with human subjects and the English language as descriptors. Other terms used included undiagnosed frequent wheezing, hygiene hypothesis, atopic sensitization, endotoxin, telemedicine, and asthma prevalence and care in rural or urban areas. STUDY SELECTION: Review and original research studies that reported on asthma care in rural areas were selected for evaluation. RESULTS: Contrary to common belief, evidence suggests that asthma prevalence of rural and urban youth is comparable in the United States and may indeed be higher in rural areas. Data are not available to determine prevalence rates for rural adults in the United States. This review presents data suggesting that the utilization of asthma care among rural and urban residents may not be comparable. Rural residents are confronted with certain barriers to care that are not as common in the urban setting. These include lack of health care insurance, geographic obstacles to obtaining care, inability of getting suitable appointments, scarcity of health care professionals, and poor access to information about asthma. CONCLUSIONS: Current literature suggests that asthma in rural areas may be underdiagnosed and deserves additional study. Improving formal and continuing education of health care professionals, compensating for health care shortages by using existing resources, and facilitating patient support and education are some of the ways to address the special needs faced by rural residents. PMID- 16312159 TI - Sedges of the family Cyperaceae are primarily in the genera Carex and Cyperus. PMID- 16312160 TI - Just say formicidae. PMID- 16312161 TI - DNA-based immunotherapy to treat atopic disease. AB - OBJECTIVE: To review the current literature regarding DNA-based immunotherapy with respect to signaling mechanisms, cytokine profiles, and the applicability and success of this strategy to treat allergic disease. DATA SOURCES: English language articles were identified from the PubMed database using both standard and clinical queries. Search terms included CpG, allergy, atopic disease, immunotherapy, DNA vaccination, immunomodulation, and immunostimulatory DNA. Other sources included bibliographies from relevant articles. STUDY SELECTION: Recent studies that provide information about the mechanisms or applications of DNA-based immunotherapy with respect to atopic disease are included in this review. RESULTS: DNA-based immunotherapy composed of unmethylated CpG repeats is capable of inducing a shift in the cytokine profile and immune response that favors the T(H)1 arm. This observation makes DNA-based immunotherapy a promising candidate for the treatment of atopic diseases, which are known to be mediated by T(H)2-based responses. Early animal and human trials of DNA-based immunotherapy have shown the strategy to be both safe and effective. CONCLUSIONS: DNA-based immunotherapy, although still in the early stages of development, has thus far been shown to be both safe and effective for a variety of atopic diseases and offers the potential for significant improvements over current immunotherapy protocols. PMID- 16312162 TI - A newborn with complete heart block and structural cardiac defects. PMID- 16312163 TI - Adverse reactions to ants other than imported fire ants. AB - OBJECTIVE: To identify ants other than Solenopsis invicta and Solenopsis richteri reported to cause adverse reactions in humans. DATA SOURCES: We conducted a literature review to identify reports of medical reactions to ants other than S. invicta and S. richteri. Our review of medical and entomological literature on stinging ants was generated from MEDLINE and FORMIS, respectively, using the key words stinging ants and ant stings. The search was limited to articles in English published from 1966 to 2004 on MEDLINE and all years on FORMIS. We also present 3 new case reports of severe reactions to stings by 2 different species of ants, Pseudomyrmex ejectus and Hypoponera punctatissima. STUDY SELECTION: Articles that concerned anaphylactic (IgE-mediated) or anaphylactic-like (resembling anaphylaxis but mechanism unknown) immediate reactions to ant stings or bites were included in this review. RESULTS: Taken together, our data demonstrate that S. invicta and S. richteri are not alone in their capability to cause serious allergic or adverse reactions. A diverse array of ant species belonging to 6 different subfamilies (Formicinae, Myrmeciinae, Ponerinae, Ectatomminae, Myrmicinae, and Pseudomyrmecinae) and 10 genera (Solenopsis, Formica, Myrmecia, Tetramorium, Pogonomyrmex, Pachycondyla, Odontomachus, Rhytidoponera, Pseudomyrmex, and Hypoponera) have now been shown to have this capability. CONCLUSION: Awareness that species other than imported fire ants may cause severe reactions should lead to more rapid evaluation and treatment and further investigation of the medical entomology of these ants. PMID- 16312164 TI - Impact of ingredient labeling practices on food allergic consumers. AB - BACKGROUND: Food allergic consumers depend on ingredient labels for allergen avoidance, and the modality of labeling is changing. OBJECTIVE: To investigate current responses to food labels so that the impact of future label changes can be anticipated. METHODS: Adults who attended Food Allergy & Anaphylaxis Network conferences completed a survey regarding their experiences with food labels for their family's most severely affected food allergic individual (FAI). RESULTS: There were 489 completed surveys (84% participation). Most FAIs were young (41% <4 years of age and 56% 4-18 years of age) and highly atopic (51% had asthma and 69% had atopic dermatitis). Food allergies included the following: peanut, 81%; tree nuts, 53%; milk, 51%; egg, 51%; and soy, 17%. All chocolate products were avoided by 37% of FAIs who were avoiding peanut and 40% who were avoiding tree nuts; 91% of tree nut allergic FAIs avoided all tree nuts. Of FAIs who avoided soy, 41% avoided soybean oil and 38% avoided soy lecithin. Of those who avoided milk, 82% avoided lactose. Allergic reactions were attributed to misunderstanding label terms (16%) and to nonspecific terms (spice, flavor) (22%). Ingredient labels were "always" or "frequently" read before purchase by 99%. Product brand choice was "very much influenced" by the manner of labeling for 86%, and manufacturers were contacted for more information by 86%. CONCLUSIONS: Our results suggest that improved product allergen labeling will reduce allergic reactions and simplify allergy management. However, the new labeling may not indicate the form or source of the allergen, and individuals who do not currently avoid foods with minimal or irrelevant protein content, such as soy oil or soy lecithin, may face additional ambiguity and unnecessary dietary restrictions. PMID- 16312165 TI - Coincidence of atopy and its profile (monosensitization/polysensitization) between sibling pairs. AB - BACKGROUND: Results of epidemiologic studies have shown that childhood atopy is probably a hereditary disorder. In the atopic population, some individuals are sensitized to only 1 class of allergens (monosensitized), whereas others are sensitized to more than 1 class of allergens (polysensitized). OBJECTIVE: To investigate whether atopy and its profile (monosensitization/polysensitization) tend to coincide in sibling pairs. METHODS: We evaluated sensitization to 5 classes of aeroallergens (house dust mites, animal danders, pollens, molds, and cockroach) using skin prick testing in 564 children with symptoms suggestive of allergic diseases (index children) and their paired siblings. RESULTS: The frequency of sibling atopy was highest (56.8%) for polysensitized index children (n=222), intermediate (45.4%) for monosensitized index children (n=196), and lowest (30.8%) for nonsensitized index children (n=146). The proportion of polysensitization among atopic siblings was significantly higher for polysensitized (47.6%) than for monosensitized (32.6%) index children. Polysensitized index children were found to more frequently have polysensitized siblings (27.0%) than were monosensitized index children (14.8%), with an odds ratio of 2.13 (95% confidence interval, 1.30-3.49), whereas the likelihood of having a monosensitized sibling was similar for monosensitized and polysensitized index children. CONCLUSION: These data suggest a coincidence of atopy and its profile in terms of monosensitization and polysensitization in sibling pairs, although the relative importance of genetic and environmental influences requires further study. PMID- 16312166 TI - Short-term tolerability of etoricoxib in patients with cutaneous hypersensitivity reactions to nonsteroidal anti-inflammatory drugs. AB - BACKGROUND: Etoricoxib is a novel cyclooxygenase 2 selective inhibitor. Until now, there has not been information in the literature about its tolerability in patients with a history of hypersensitivity reactions to nonsteroidal anti inflammatory drugs (NSAIDs). OBJECTIVE: To determine the short-term tolerability of etoricoxib in patients with a history of cutaneous adverse reactions to NSAIDs. METHODS: Single-blind challenge testing was performed on 2 different days using placebo (talc) and etoricoxib. On the first day, 2 placebo capsules were administered 1 hour apart; 7 days later, each patient received divided doses of the total therapeutic dose of 90 mg of etoricoxib: 22.5 mg initially and 67.5 mg 1 hour later if no reactive symptoms were noted. RESULTS: Of 141 patients who underwent challenge testing with etoricoxib, only 2 (1.4%) had positive test results; both developed wheals on the extremities. These 2 patients were treated with chlorpheniramine maleate (10 mg intravenously), and the symptoms completely resolved within 2 hours. None of the patients experienced adverse reactions to the placebo challenge. CONCLUSION: The low rate of adverse reactions to etoricoxib, tested by oral challenge, suggests that patients with previous cutaneous hypersensitivity reactions to NSAIDs (primarily urticaria and angioedema) may tolerate this drug. PMID- 16312167 TI - Effects of omalizumab and budesonide on markers of inflammation in human bronchial epithelial cells. AB - BACKGROUND: Many patients with asthma have an IgE-mediated allergic component to the disease. Omalizumab, a monoclonal anti-IgE antibody, has demonstrated clinical efficacy in patients with allergic asthma. The effects of omalizumab on inflammation in asthma are not completely understood. OBJECTIVES: To evaluate the effects of omalizumab on allergen- and growth factor-stimulated proinflammatory cytokine and nitric oxide (NO) production in human bronchial epithelial cells (BECs) and to compare them to the effects of budesonide, a corticosteroid with known anti-inflammatory properties. METHODS: Human BECs were stimulated in duplicate with interleukin 1beta (IL-1beta), 100 U/mL; ragweed, 10 microg/mL; dust mite, 1000 AU; and epithelial growth factor, 40 ng/mL; and either 10(-7) M budesonide or 0.1 microg/mL of omalizumab in a 4% dust mite atopic serum medium for 6 and 24 hours in 5% carbon dioxide at 37 degrees C. Tumor necrosis factor alpha and transforming growth factor betaexpression and production and IL-4, IL 13, and NO production were assayed using gene-specific messenger RNA or sensitive enzyme-linked immunosorbent assays. RESULTS: Omalizumab inhibited the expression and of production proinflammatory cytokines and growth factor in antigen stimulated BECs at 6 and 24 hours. Production of NO was inhibited at 6 hours and increased at 24 hours by omalizumab and budesonide. CONCLUSIONS: The effects of omalizumab were similar to those of budesonide. These results, consistent with previously reported evidence of anti-inflammatory effects of omalizumab, demonstrate that omalizumab may reduce airway inflammation and probably contributes to decreased airway remodeling in patients with asthma. PMID- 16312168 TI - Effect of fluticasone propionate-salmeterol therapy on seasonal changes in airway responsiveness and exhaled nitric oxide levels in patients with pollen-induced asthma. AB - BACKGROUND: There has been concern that in allergic asthmatic patients there might be an interactive effect on inflammation between regular salmeterol use and exposure to allergens, resulting in increased airway responsiveness. OBJECTIVE: To determine the effects of salmeterol on allergen-induced changes in airway responsiveness and exhaled nitric oxide (ENO) levels in allergic asthmatic patients concomitantly taking inhaled corticosteroids. METHODS: Forty-two asthmatic patients sensitized to pollen allergens were randomly allocated to treatment with fluticasone propionate-salmeterol (n=21) or fluticasone propionate alone (n=21). Spirometry, the methacholine provocation concentration causing a 20% decline in forced expiratory volume in 1 second (PC20), the adenosine 5' monophosphate (AMP) PC20, and ENO levels were measured before and at the height of the pollen season after 6 weeks of treatment. RESULTS: Changes in the methacholine PC20, the AMP PC20, and ENO levels were not significantly different between treatment groups. No significant changes in the AMP PC20 were observed among the fluticasone propionate-salmeterol and fluticasone propionate groups during natural pollen exposure. However, a significant increase in the methacholine PC20 was observed in the fluticasone propionate-salmeterol group (P = .03) and in the fluticasone propionate group (P = .04); ENO concentrations decreased significantly in both groups during natural allergen exposure (P = .009 and .005). CONCLUSIONS: In patients with pollen-induced asthma, treatment with either fluticasone propionate or fluticasone propionate-salmeterol is associated with significant reductions in methacholine responsiveness and ENO concentrations, even during natural pollen exposure. Furthermore, at least in patients with mild asthma, natural allergen exposure and the regular use of fluticasone propionate-salmeterol are not associated with a greater increase in ENO levels and airway responsiveness than natural allergen exposure and fluticasone propionate use alone. PMID- 16312169 TI - Characterization and improvement of apple extracts for the diagnosis of apple IgE mediated allergy. AB - BACKGROUND: The production process of reliable fruit extracts is not well established. OBJECTIVES: To improve the overall quality of apple extracts by reducing protein loss during the manufacturing process and to evaluate the improved extracts using in vivo and in vitro experiments. METHODS: Two types of extracts were prepared from peels of Golden Delicious apples (Malus domesticus). Extract A was extracted, 1:2 wt/vol, for 30 minutes at 40 degrees C in 0.01 M phosphate-buffered saline, and extract B was extracted, 1:2 wt/vol, in phosphate buffered saline with 20% polyvinylpolypyrrolidone and 2-mmol/L EDTA. Both extracts were filtered, dialyzed in 3.5-kDa dialysis membranes, and lyophilized. The antigenic and allergenic profiles were analyzed using immunoblot and enzyme linked immunosorbent assay. Nine patients clinically sensitive to apples and 12 controls underwent skin testing with both extracts. RESULTS: Extracts A and B had dry weight yields of 0.71% and 1.86% and protein contents of 104.6 and 257 microg/mg of freeze-dried material, respectively. A steady and progressive loss of protein, greater in extract A than in extract B, was detected at different intervals during the manufacturing process of both extracts. Extract B produced larger wheal sizes than extract A (P = .008). Enzyme-linked immunosorbent assay inhibition results confirmed that extract B had a greater inhibition capacity than extract A. CONCLUSIONS: A progressive loss of protein content occurs during the manufacturing of apple extracts. Wheal sizes induced by extract B were significantly larger than those induced by extract A and prick-by-prick solutions. Extract B was also more potent in vitro than extract A. PMID- 16312170 TI - An inverse relationship between peroxisome proliferator-activated receptor gamma and allergic airway inflammation in an allergen challenge model. AB - BACKGROUND: Peroxisome proliferator-activated receptor gamma (PPAR-gamma) expression has not been evaluated in bronchoalveolar lavage (BAL) cells from allergic asthmatic patients. OBJECTIVE: To determine whether inappropriate down regulation of PPAR-gamma in alveolar macrophages may contribute to persistent airway inflammation in allergic asthma. METHODS: We used segmental allergen challenge as a model of in vivo experimental allergic asthmatic exacerbation and airway inflammation. PPAR-y gene expression was evaluated at baseline and 24 hours later in asthmatic patients and controls using real-time polymerase chain reaction. Immunofluorescence was used to determine cellular location of the PPAR gamma protein. RESULTS: We demonstrate for the first time to our knowledge that PPAR-gamma messenger RNA and protein, which are highly expressed in alveolar macrophages of healthy individuals, are significantly reduced in asthmatic patients after segmental allergen challenge. In allergic asthmatic patients (n=9), PPAR-gamma gene expression decreased significantly from baseline to postchallenge BAL (median decrease, 45%; P = .008). Furthermore, immunofluorescence staining demonstrated that PPAR-gamma protein was associated with alveolar macrophages and not with inflammatory eosinophils and neutrophils. CONCLUSION: Results implicate down-regulation of PPAR-gamma in BAL cells as a potential factor in dysregulation of lung homeostasis in asthmatic patients. The present findings suggest that PPAR-gamma agonists could have a future role in asthma therapy and warrant further study. PMID- 16312171 TI - Safety and efficacy of olopatadine hydrochloride nasal spray for the treatment of seasonal allergic rhinitis to mountain cedar. AB - BACKGROUND: A nasal spray containing the antiallergy agent olopatadine hydrochloride is being developed for the treatment of seasonal allergic rhinitis (SAR) to mountain cedar. OBJECTIVE: To evaluate the safety and efficacy of 2 concentrations of olopatadine nasal spray vs placebo nasal spray in patients with SAR to mountain cedar. METHODS: This was a multicenter, randomized, double-blind, placebo-controlled study. After a 3- to 21-day placebo run-in, 677 patients aged 12 to 81 years were randomized to receive 0.4% or 0.6% olopatadine or placebo, 2 sprays per nostril twice daily for 2 weeks. Patients evaluated morning and evening reflective and instantaneous nasal symptoms (sneezing, stuffy nose, runny nose, and itchy nose, which compose the total nasal symptom score [TNSS]) and ocular symptoms. RESULTS: Olopatadine spray (0.4% and 0.6%) was statistically significantly superior to placebo for percentage change from baseline in overall reflective and instantaneous TNSSs. Also, 0.6% olopatadine was statistically significantly superior to placebo for reducing the reflective and instantaneous assessments of sneezing, runny nose, itchy nose, stuffy nose, itchy eyes, and watery eyes. Olopatadine spray exhibited a safety profile comparable with that of placebo. CONCLUSIONS: Olopatadine nasal spray (0.4% and 0.6%) provided statistically significant improvements in allergic rhinitis symptoms compared with placebo regarding TNSSs and individual symptoms, including congestion, itchy and runny nose, sneezing, and itchy and watery eyes, in patients with SAR to mountain cedar. Olopatadine nasal spray administered twice daily was safe and well tolerated in adolescents and adults. PMID- 16312172 TI - Cord blood house dust mite allergen in newborns: relationship to maternal blood levels of allergen and allergen specific IgG and IgE. AB - BACKGROUND: House dust mite allergen has previously been detected in the cord blood of some newborns but not others. The factors that affect the presence and levels in cord blood of this and possibly other allergens are unknown. OBJECTIVE: To test the hypothesis that the levels of maternal allergen and allergen specific IgG affect the presence and levels of newborn allergen. METHODS: Enzyme-linked immunosorbent assays were used to measure the levels of house dust mite allergen Der p 1 and Der p 1 specific IgG and IgE in paired blood samples from 98 mothers and full-term newborns. RESULTS: Der p 1 was detected in 27 mothers and 12 newborns. None of the 71 mothers who lacked Der p 1 had Der p 1-positive newborns. When present, cord blood Der p 1 levels correlated with and were approximately one third of Der p 1 levels in maternal blood. Cord blood Der p 1 levels also tended to correlate with maternal blood levels of Der p 1 specific IgG but showed little, if any, correlation with maternal blood levels of Der p 1 specific IgE. CONCLUSIONS: When detectable, the levels of an allergen in the blood of newborns correlate with the levels of that allergen in the blood of their mothers and tend to be related to maternal levels of allergen specific IgG. PMID- 16312173 TI - Fatal hypersensitivity pneumonitis. AB - BACKGROUND: Hypersensitivity pneumonitis (HP) is an uncommon, non-IgE-mediated interstitial lung disease caused by the inhalation of a variety of organic dusts, most commonly from exposure at work or in the pursuit of hobbies. Typically, after the disease is recognized, the causative allergen or environment is identified and treatment initiated through avoidance measures and corticosteroids. Progression of the disease is then usually halted and even reversed. Fatal cases of HP are unusual. OBJECTIVE: To report a case of progressive and deadly HP in a 40-year-old printer who developed subacute bird fancier's disease with its clinical characteristics and positive precipitins to pigeon proteins. METHODS: Chest x-ray examinations and tests of lung function were performed in the patient. Two months after initial consultation, when the diagnosis was still elusive, an open lung biopsy was performed and the patient was treated with prednisone for 3 months. A subsequent chest x-ray examination was performed 4 months after the biopsy. RESULTS: With avoidance of birds and treatment with corticosteroids, the patient's symptoms resolved and lung function normalized. He was subsequently diagnosed as having asthma followed by bronchitis and 2 episodes of pneumonia. He did not fully recover from these but developed progressive dyspnea. After linking his symptoms to work by history, he underwent lung biopsy with findings consistent with chronic HP. Serum antibody titers were positive for Aspergillus but not pigeon proteins. Based on exposure to water based coolants, he was suspected of having chronic occupational HP, although this could not be confirmed. Despite aggressive treatment, he developed a progressive course that was ultimately fatal. CONCLUSIONS: This report details the progressive disease course in an individual who presented initially with subacute HP. Unfortunately, even after appropriate diagnosis and management, the course of the disease can be fatal. PMID- 16312174 TI - Allergic bronchopulmonary aspergillosis in a patient with cystic fibrosis: diagnostic criteria when the IgE level is less than 500 IU/mL. AB - BACKGROUND: Recently, the Cystic Fibrosis Foundation developed a consensus report recommending diagnostic criteria for allergic bronchopulmonary aspergillosis (ABPA) in patients with cystic fibrosis that includes a serum IgE level greater than 500 IU/mL as the "minimal diagnostic criterion." OBJECTIVE: To describe a 7 year-old girl with ABPA whose serum IgE level increased to only 398 IU/mL. METHODS: Total IgE and anti-Aspergillus serologic measurements were performed using enzyme-linked immunosorbent assay and standard laboratory techniques; HLA analysis was performed; interleukin 4 receptor alpha single nucleotide polymorphisms were performed using polymerase chain reaction and DNA sequencing; CD23+ B cells were measured using flow cytometry; and cytokine synthesis to Aspergillus purified antigens was assessed using flow cytometry. RESULTS: A 7 year-old girl with cystic fibrosis who had mild pulmonary disease and well controlled asthma developed pulmonary infiltrates, increased wheezing, and decreased pulmonary function. Additional studies demonstrated peripheral blood eosinophilia (eosinophil count, 1807 cells/mm3 [19%]) and an increase in IgE and IgG anti-Aspergillus serology; bronchoalveolar lavage revealed septate hyphae with 45 degrees branching subsequently identified as A fumigatus and pulmonary eosinophilia. Previous HLA typing revealed that the patient was HLA-DR2+, DRB*1501, HLA-DQ2-, a pattern associated with increased risk of ABPA. In addition, there was increased up-regulation of CD23 molecules by interleukin 4 stimulation on the patient's B cells, as observed in ABPA. The patient was treated with corticosteroids and itraconazole with resolution of symptoms and pulmonary infiltrates. CONCLUSIONS: Examination of the pulmonary inflammatory response using bronchoalveolar lavage, genetic risk with HLA-DR2+DQ2- typing, and increased interleukin 4 sensitivity are useful adjunctive studies in the diagnosis of ABPA. PMID- 16312175 TI - Are specific antiretrovirals associated with an increased risk of discontinuation due to toxicities or patient/physician choice in patients with hepatitis C virus coinfection? AB - BACKGROUND: Liver damage associated with hepatitis C (HCV) may influence the likelihood of experiencing discontinuation due to toxicities or patient/physician choice (TOXPC) in patients taking combination antiretroviral therapy (cART). Little information to address this concern is available from clinical trials as patients with HCV are often excluded. AIMS: To compare incidence rates of discontinuation due to TOXPC associated with specific antiretrovial drugs in patients with or without HCV. PATIENTS/METHODS: A total of 4929 patients from EuroSIDA under follow-up from January 1999 on a specific nucleoside pair (zidovudine/lamivudine, didanosine/stavudine, stavudine/lamivudine, or other) with a third drug (abacavir, nelfinavir, indinavir, nevirapine, efavirenz, lopinavir/ritonavir or other boosted-protease inhibitor (PI)-containing regimen) and with known HCV serostatus were studied for the incidence of discontinuation of any nucleoside pair or third drug due to TOXPC. Incidence rate ratios were derived from Poisson regression models. RESULTS: In total 1358 patients had HCV (27.5%). During 12 799 person-years of follow-up there were 2141 discontinuations due to TOXPC for nucleoside pairs and 2501 for third drugs. The incidence of discontinuation due to TOXPC was consistently higher in patients with HCV after stratification by nucleoside pair or third drug. After adjustment for CD4+ count, gender, exposure group, time on HAART, region and treatment regimen, there were few differences in the rate of discontinuation due to TOXPC in those with HCV compared with those without for any nucleoside pairs or third drugs. Similar results were seen when concentrating on discontinuation due to toxicities alone. CONCLUSIONS: Although patients with HCV generally had higher rates of discontinuation due to TOXPC compared with patients without HCV, there was little evidence to suggest that this was associated with any specific nucleoside pair or third drug used as part of cART. Our results do not suggest that any specific component of cART is more poorly tolerated in patients with HCV or that the presence of HCV should influence the choice between antiretrovirals used as part of a cART regimen. PMID- 16312176 TI - Thymidine analogue mutation profiles: factors associated with acquiring specific profiles and their impact on the virological response to therapy. AB - BACKGROUND: Studies have suggested that HIV-1 may develop thymidine analogue mutations (TAMs) by one of two distinct pathways - the TAM1 pathway (including mutations 41L, 210W and 215Y) or the TAM2 pathway (including mutations 67N, 70R and 219E/Q) - under the pressure of a not fully suppressive thymidine-analogue containing regimen. METHODS: Frozen plasma samples stored in the EuroSIDA repository were selected and sent to two central laboratories for genotypic analysis. We considered 733 patients with at least one genotypic test showing > or =1 TAMs (the first of these tests in chronological order was used). TAM1 and TAM2 genotypic profiles were defined in accordance with previous literature. Statistical modelling involved logistic regression and linear regression analysis for censored data. RESULTS: The observed frequencies of patterns classifiable as TAM1 or TAM2 profiles were markedly higher than the probabilities of falling into these classifications by chance alone. The chance of detecting a TAM2 profile increased by 25% per additional year of exposure to zidovudine. We found that mutations 67N and 184V were not associated with a particular TAM profile. In the presence of TAM2 profiles, the adjusted mean difference in the 6-month viral reduction was 0.96 log10 copies/ml (95% confidence interval: 0.20; 1.73) higher in patients who started stavudine-containing regimens instead of zidovudine containing regimens. CONCLUSIONS: This study provides evidence that the suggested TAM clustering is a real phenomenon and that it may be driven by which thymidine analogue the patients has used. In patients with TAM2-resistant viruses, stavudine appears to retain greater viral activity than zidovudine. PMID- 16312177 TI - Saquinavir 500 mg film-coated tablets demonstrate bioequivalence to saquinavir 200 mg hard capsules when boosted with twice-daily ritonavir in healthy volunteers. AB - OBJECTIVE: To establish the bioequivalence of a 500 mg film-coated tablet of saquinavir mesylate (FCT SQV) to the 200 mg hard-capsule saquinavir mesylate (HC SQV), both boosted with ritonavir and administered under fed conditions. METHODS: We carried out a multi-centre, open-label, randomized, two-sequence, four-period, two-treatment, replicated crossover study in 93 healthy men and 7 healthy women. Individuals were randomly assigned to receive sequential single doses of saquinavir in one of two treatment sequences: ABAB or BABA. Individuals received 100 mg ritonavir twice daily for 24 days. On days 14,17, 20 and 23, study participants took 1000 mg of HC SQV (five 200 mg capsules, treatment A) or FCT SQV (two 500 mg tablets, treatment B) with a high-fat, high-calorie breakfast, and pharmacokinetic analyses were carried out over the next 24 hours. Area under the saquinavir concentration-time curve (AUC0-alpha), maximum saquinavir plasma concentration (Cmax), time to Cmax and terminal half-life were calculated. The relative bioavailability of FCT SOV versus HC SQV was calculated as the ratio of the respective estimated mean saquinavir AUC0-alpha and Cmax. The calculation was based on an ANOVA including the factors site, sex, sequence, period, treatment and study participant to the log-transformed parameters log(AUC0-alpha) and log(Cmax); the relative bioavailability and the 90% confidence intervals (CIs) were estimated using the treatment contrasts of the ANOVA. Bioequivalence was concluded as for both parameters, AUC0-alpha and Cmax, the 90% CIs for the relative bioavailability were entirely included in the reference region [0.80 1.25]. RESULTS: Saquinavir plasma concentration-time profiles for the two formulations were similar. Geometric mean AUC0-alpha and Cmax values were clearly increased for FCT SQV (26 826 versus 24 430 h*ng/ml; and 3644 versus 3064 ng/ml, respectively); ratios of mean exposures were estimated to be 1.10 for AUC0-alpha and 1.19 for Cmax of saquinavir. However, the corresponding two-sided 90% CIs (1.04-1.16 and 1.14-1.25, respectively) all fell within the limits set for equivalence (0.80, 1.25). The adverse event profile for FCT SQV was similar to that for HC SQV. CONCLUSION: The new 500 mg FCT SQV formulation is bioequivalent to the 200 mg HC SQV formulation, at the dose of 1000 mg, in combination with 100 mg ritonavir under fed conditions. The 500 mg FCT SQV formulation reduces pill count for boosted saquinavir (SQV/r) from six capsules to three tablets twice daily. This may increase patient acceptability of SQV/r, particularly in less treatment-experienced patients. PMID- 16312178 TI - Simplification therapy with once-daily didanosine, tenofovir and efavirenz in HIV 1-infected adults with viral suppression receiving a more complex antiretroviral regimen: final results of the EFADITE trial. AB - BACKGROUND: High pill burden and side effects often impact on the long-term success of highly active antiretroviral therapy (HAART), which has led clinicians to search for more convenient regimens. PATIENTS AND METHODS: A prospective, multicentre, open, comparative study in which HIV-1-infected patients on HAART and with plasma HIV-1 RNA <50 copies/ml for longer than 6 months were switched to tenofovir, didanosine and efavirenz (QD arm) or remained on the same treatment regimen (control arm). Patients with grade 4 toxicities or plasma HIV-1 RNA values repeatedly >1000 copies/ml discontinued the study. RESULTS: A total of 390 patients were included in the trial (309 in the QD arm and 81 in the control arm). The main baseline characteristics were well balanced between groups. In the QD arm, 41% of patients received high (standard) didanosine doses and 59% received reduced doses. At 12 months, plasma HIV-1 RNA <400 copies/ml was attained in 66% of QD patients and 73% of controls in the intent-to-treat (ITT) analysis (P=NS). However, the number of individuals with HIV-1 RNA <400 copies/ml in the QD arm was 56% versus 71% when comparing the use of high versus low didanosine doses (P=0.007). Treatment discontinuation occurred in 87 QD cases (28%) and 17 controls (21%). Twenty QD individuals (6.5%) and 2 controls (2.5%) discontinued because of virological failure (P=NS). The median CD4+ cell count change at 12 months was -26 and +27 cells/microl in QD patients and controls, respectively (P=0.001). In individuals who attained HIV-1 RNA <400 copies/ml, CD4+ cell changes were -25 and +15 cells/microl in QD patients and controls, respectively (P=0.001). Moreover, CD4+ cell declines in the QD arm were significantly greater in patients taking high versus low didanosine doses (-59 versus -15 cells/microl; P=0.04). The lipid profile improved significantly in the QD arm, particularly in patients who were on protease inhibitors prior to simplification. CONCLUSIONS: Simplification to didanosine-tenofovir-efavirenz provides a virological suppression rate at 12 months similar to that seen in patients who do not change therapy, as long as low didanosine doses are administered. Decreases in CD4+ cell levels in patients in the QD arm (especially decreases seen with high didanosine doses) and dyslipidaemias along with less convenient pill burden and schedules in controls were the main long-term concerns for each option. PMID- 16312179 TI - Design and characterization of human respiratory syncytial virus entry inhibitors. AB - Human respiratory syncytial virus (hRSV) is a pathogen of worldwide health concern. The crucial membrane fusion event during viral entry into host cells involves a 'trimer-of-hairpins' structure that brings the amino (N)- and carboxy (C)-terminal regions of the viral fusion glycoprotein (F protein) into close proximity. Two heptad repeat regions that are highly conserved in the F protein - HR1 (N-terminal) and HR2 (C-terminal) - have an important role in this process. It has been shown that both HR1-and HR2-based peptides can inhibit viral entry. However, these proteins, and the HR1 peptides in particular, are liable to aggregation. We designed three peptides containing multiple copies of alternating HR1 and HR2 sequences denoted 5-Helix, HR121 and HR212, respectively. The 5 Helix, HR121 and HR212 proteins were functionally analogous to single HR1, HR1 and HR2 sequences, respectively. All three proteins were expressed in soluble form and biophysical analysis showed that they exhibited alpha-helical secondary structures. The three proteins were potent fusion inhibitors in vitro, at the micromolar scale, with the HR1 analogues being approximately two times more effective than the HR2 analogue. Our results suggest that these rationally designed protein inhibitors could serve as a new class of anti-hRSV agents. PMID- 16312180 TI - Short communication. Baseline factors associated with haematological toxicity that leads to a dosage reduction of pegylated interferon-alpha2a and ribavirin in HIV- and HCV-coinfected patients on HCV antiviral therapy. AB - OBJECTIVE: To assess the baseline factors associated with haematological toxicity that lead to ribavirin or pegylated interferon (peginterferon) dosage reductions in hepatitis C and human immunodeficiency virus (HCV/HIV)-coinfected patients. DESIGN: Multicentre, prospective, observational study. SETTING: Eleven hospitals in Spain during the period 2002-2003. SUBJECTS AND METHODS: One-hundred and forty two HIV/HCV-coinfected patients received peginterferon-alpha2a plus ribavirin. Baseline characteristics and haematological parameters were recorded at baseline, week 4, 8, 12, 24 and 48. Cox's regression model was used to study the factors associated with the appearance of a haemoglobin level below 10g/dl (haemoglobin endpoint), a neutrophil count below 750/mm(3) (neutrophil-endpoint) and a platelet count below 50,000/mm(3) (platelet-endpoint). RESULTS: Nineteen patients (13.4%) reached the haemoglobin-endpoint, 22.5% the neutrophil-endpoint and 7% the platelet-endpoint. Mean time of follow-up was 8 months (+/-3.5). A baseline haemoglobin level below 14g/dl [hazard ratio (HR): 3.65; 95% confidence interval (CI): 1.46-9.06] and treatment with zidovudine (HR: 3.25; 95% CI: 1.31-8.11) were the independent factors associated with the appearance of the haemoglobin endpoint. A baseline neutrophil below 2050/mm(3) (HR: 3.59; 95% CI: 1.77-7.28) and baseline weight <60 kg (HR: 2.21; 95% Cl: 1.04-4.56) were independently associated with the appearance of the neutrophil-endpoint. Baseline platelet count (x1000/mm(3) decrease) (HR: 1.074; 95% CI: 1.04-1.11) was independently associated with the appearance of the platelet-endpoint. CONCLUSIONS: Baseline factors allow the identification of a subset of HIV/HCV-coinfected patients who are prone to experience haematological toxicity during HCV antiviral therapy. PMID- 16312181 TI - Short communication. Association of the CCR5delta32 mutation with clinical response and >5-year survival following initiation of first triple antiretroviral regimen. AB - OBJECTIVE: The CCR5delta32 mutation is associated with slower HIV disease progression in untreated infection. However, it remains controversial as to whether CCR5delta32 is a relevant prognostic marker in the context of highly active antiretroviral therapy (HAART). Here we investigate associations between CCR5delta32 and HAART outcomes in a large, population-based cohort of >1000 antiretroviral-naive individuals initiating triple therapy over a median >5 year follow-up. METHODS: CCR5delta32 genotypes were determined using PCR and DNA sequencing in a cohort of 1188 antiretroviral-naive individuals initiating triple therapy in British Columbia. Associations between CCR5delta32 and baseline (pre therapy) sociodemographic and clinical parameters were investigated in a cross sectional analysis. Cox proportional hazards regression was used to investigate the effect of CCR5delta32 on clinical outcomes following therapy initiation. The endpoints that were evaluated included time to plasma viral load (pVL) suppression <400 copies HIV RNA/ml, subsequent time to pVL rebound > or =400 copies/ml, time to CD4+ cell decline below baseline, and time to non-accidental death over a median >5 year follow-up. RESULTS: CCR5delta32 genotypes were available for 1174 of 1188 individuals (98.8%): 171 (14.6%) CCR5wt/delta32 and 1003 (85.4%) CCR5wt/wt. At baseline, CCR5wt/delta32 individuals had higher CD4+ cell counts (P=0.04), lower plasma viral loads (P=0.06) and were slightly older than CCR5wt/wt individuals (P=0.04). In multivariate analyses controlling for baseline parameters and adherence estimates, we observed a significant association between CCR5wt/delta32 and a shorter time to initial pVL suppression <400 copies/ml (multivariate hazard ratio [HR]: 1.20; 95% confidence interval [CI]: 1.01-1.44). No associations were observed between CCR5delta32 and other clinical outcomes including subsequent time to pVL rebound or time to CD4+ cell decline below baseline. In univariate analyses, we observed a significant association between CCR5wt/delta32 genotype and improved survival over the median >5 year period following initiation of HAART (P=0.03). However, this association did not remain significant in multivariate analyses after adjusting for baseline factors including adherence (multivariate HR: 0.64; 95% CI: 0.38-1.07; P=0.09) CONCLUSION: Results indicate that, after controlling for adherence, the CCR5delta32 mutation is likely not a clinically significant predictor of longer term clinical responses or survival in the context of HAART. PMID- 16312182 TI - Short communication. Antiretroviral drug resistance among drug-naive HIV-1 infected individuals in Djibouti (Horn of Africa). AB - OBJECTIVE: To survey the frequency of genotypic antiretroviral resistance in drug naive HIV-1-infected Djiboutians. METHODS: A national study was conducted in the general population of Djibouti in March 2002 to determine HIV-1 seroprevalence. Blood samples were collected anonymously and plasma samples scoring positive for HIV-1 antibodies were tested for viral load. Genotypic studies were performed with viral RNA from plasma using the consensus technique of the Agence Nationale de Recherche sur le SIDA (www.hivfrenchresistance.org). Mutations were identified using the International AIDS Society-USA resistance panel and resistant virus was defined according to the ANRS algorithm. RESULTS: A panel of 2423 individuals representing the general population of Djibouti was included. Antibodies were detected in 53 of 2423 samples tested. The HIV-1 seroprevalence in the general population was 2.2%. Genotype C was the most prevalent, and the other isolates were CRF_02 AG, or subtype A or D. Forty-seven of the 53 samples were tested for genotypic resistance, and mutations concerning all three classes of antiretrovirals were found. The most frequent were secondary mutations associated with protease inhibitors (PIs): M36I, R41K and K20I/R. A few strains displayed primary mutations (the non-nucleoside reverse transcriptase inhibitor [NNRTI] associated mutations K101E, K103T, L100I and G190V; the PI-associated mutation N88D; and the NRTI-associated mutation K65R). The presence of these mutations may be due to the transmission of strains from treated patients. CONCLUSION: Substantial polymorphism and a few primary mutations are found in HIV-1 non-B subtype isolates from Djiboutian antiretroviral-drug-naive individuals. This needs to be taken into account to adapt antiretroviral regimens and prophylactic schedules locally. PMID- 16312183 TI - Letter. In vitro phenotypic susceptibility to nucleoside reverse transcriptase inhibitors of HIV-2 isolates with the Q151M mutation in the reverse transcriptase gene. AB - In HIV-2 infection, many studies have reported a high frequency of selection of the Q151M mutation, but its impact on phenotypic susceptibility of HIV-2 isolates remains unclear. Four HIV-2 infected patients from the French ANRS HIV-2 cohort, with evidence of Q151M mutation in both plasma and available peripheral blood mononuclear cells (PBMCs) co-cultivated supernatants, were selected. In vitro phenotypic susceptibilities to different nucleoside reverse transcriptase inhibitors (NRTIs) were determined using a PBMC assay. In HIV-2 isolates, the Q151M mutation alone impacts only the phenotypic susceptibility to stavudine and abacavir. A decrease in susceptibility to all NRTIs was observed when Q151M was selected with V111I, a mutation of unknown impact on HIV-1 resistance. Clinical relevance of these phenotypic susceptibility results needs to be evaluated in HIV 2 treated patients. PMID- 16312184 TI - Opinion paper. Resistance to new anti-HIV agents: problems in the pathway of drug registration. AB - Resistance data are now requested by the regulatory agencies as an integral part of the approval process of new antiretroviral drugs. We examined the means by which resistance data was gathered during pre-clinical and clinical Phases I, II and III of drug development, and how the public and academic experts access these proprietary data. The analysis identified various opportunities for improvement of the current process, in particular the need for standards in generating and reporting resistance data on new antiretroviral drugs, and the need to enforce warnings in the product labelling on the drug combinations that can potentially lead to resistance and treatment failure. PMID- 16312185 TI - Paediatric critical care: beyond the walls. PMID- 16312186 TI - Transfer of head-injured patients in Jamaica: is there a problem? AB - Head-injured patients are often transferred to the University Hospital of the West Indies (UHWI) for tertiary care. There is no standardized, agreed protocol governing their transfer. During the three-year period January 1998 to December 2000, 144 head injured patients were transferred to the UHWI from other institutions. They were 70% male, had a mean age of 34 years and spent a mean of 13 days in hospital. Eighteen per cent were admitted to the Intensive Care Unit, where they spent a mean of nine days. On arrival, mean pulse rate was 92 +/- 22 beats/minute, mean systolic blood pressure was 130 +/- 27 mmHg and mean diastolic was 76 +/- 19mmHg. Twenty-eight per cent of patients had a pulse rate above 100/min on arrival and 13.8% had systolic blood pressure below 60 mmHg. The Glasgow Coma Scale score was unrecorded at the referring institution in 70% of cases and by the receiving officers at the UHWI in 23% of cases. Intubation was done on only half of those who were eligible. Junior staff members initiated and carried out transfers whenever this was documented. The types of vehicles and monitoring equipment used could not be determined in most instances. Fifty-eight per cent of patients had minor head injuries, 12%, severe injury and 33%, associated injuries requiring a variety of surgical procedures by multiple specialties. Most patients (80.6%) were discharged home but 11.8% died in hospital. Transfer of head-injured patients, many with multiple injuries is not being performed in a manner consistent with modern medical practice. There is urgent need for implementation of a standardized protocol for the transfer of such patients in Jamaica. PMID- 16312187 TI - Microbial isolates from patients in an intensive care unit, and associated risk factors. AB - A retrospective one-year analysis of blood, sputum and urine samples taken from all patients admitted for more than 48 hours to the Intensive Care Unit at the University Hospital of the West Indies (UHWI) was undertaken. Positive trapped sputum cultures were found in 50% of patients, positive blood cultures in 32.7% and positive urine cultures in 23.1%. Gram-negative organisms predominated especially Pseudomonas aeruginosa (41.3%) and Acinetobacter spp (33.5%). Coagulase-negative staphylococcus (20%) and streptococcus group D (18.7%) were the most common gram-positive organisms. The Acinetobacter spp showed marked resistance to most antibiotics except for meropenem (82.7% susceptibility) while P. aeruginosa was most susceptible to ceftazidime (84.4%) and amikacin (89.1%). Both the coagulase-negative staphylococcus and streptococcus group D were relatively sensitive to amoxycillin/clavulanate (80.6% and 79.3% respectively). There was a high incidence of yeast found in sputum (27.1%) and urine (16.8%). Mechanical ventilation was a significant risk factor for developing a positive sputum culture (p = 0.01), this effect being particularly prominent in those ventilated for > 5 days. Central venous pressure lines significantly increased the risk of a positive blood culture (p = 0.005). This increase was seen particularly in those with CVP lines for > 7 days. Other risk factors for developing positive cultures included preadmission infection, antibiotic use just prior to ICU admission, increasing APACHE II score and increasing age. PMID- 16312188 TI - The effect of Hurricane Ivan on Emergency Department operations at the University Hospital of the West Indies. AB - The objective of this study was to determine the impact of Hurricane Ivan on the operations of the Emergency Department (ED) at the University Hospital of the West Indies (UHWI). Hurricane Ivan,--a category four Hurricane--traversed the south coast of the island of Jamaica on September 10, 2004, causing damage to several parishes. The study design is a descriptive retrospective record-based incidence study of all admissions to the ED commencing at 8:00 am September 10, 2004 and ending at 12 midnight September 11, 2004, covering the period of the entire duration of the hurricane as well as immediately after. Injuries that took place during the hurricane, inclusive of gun-shot wounds (GSWs) represented 40% of the total patients seen. The types of admission are listed categorically as well as specific items to be considered in planning for potential disasters of higher magnitude. Overall, the coping mechanism of the ED at UHWI was adequate for this magnitude of disaster. PMID- 16312189 TI - The sero-epidemiology of measles in children from Eastern Turkey. PMID- 16312190 TI - Judet quadricepsplasty for extension contracture of the knee. AB - Extension contracture of the knee is a well known complication of severe femoral fractures, especially in the supracondylar region. Traditional management by the Thompson quadricepsplasty may result in a variable return of knee flexion and the possibility of significant extension lag. The Judet technique of quadricepsplasty offers the advantages of a controlled, sequential release of the intrinsic and then the extrinsic components limiting knee flexion and a reduced potential for iatrogenic quadriceps rupture or extension lag. The modified Judet quadricepsplasty has definite advantages over the Judet technique since it usually involves less soft tissue dissection and consequently less blood loss. PMID- 16312191 TI - Carcinoma of the jejunum with multideposit peritoneal seeding, resection and intraperitoneal chemotherapy. AB - Jejunal adenocarcinoma is rare, often presenting late with widespread intraperitoneal disease. Intraperitoneal chemotherapy (IPC) has been shown in non randomized studies to improve the survival of patients presenting with intraperitoneal metastases from carcinoma of the colon, appendix and stomach and in primary peritoneal malignancies including mesothelioma and pseudomyxoma peritonei, providing that adequate operative cytoreduction can be performed. A case is presented of obstructive jejunal adenocarcinoma in which 19 intraperitoneal deposits were excised. The patient was treated successfully with immediate postoperative IPC followed by systemic chemotherapy. This condition is reviewed along with the rationale for IPC. PMID- 16312192 TI - Penetrating type intraosseous ganglion cyst of the lunate bone. AB - Intraosseous ganglion cysts of the carpal bones located in the lunate are one of the rarely seen pathologic conditions. Here we present a case of the penetrating type of intraosseous ganglion cyst located in the lunate as an uncommon cause of wrist pain. The patient was successfully treated by surgical intervention. Intraosseous ganglion cyst should be considered in the differential diagnosis of chronic dull wrist pain. PMID- 16312193 TI - An in vitro comparison of implant materials cell attachment, cytokine and osteocalcin production. AB - Bone deposition, for any implant system, is the deciding factor for the success. The biochemical signals at the cellular level will help elucidate the direction of host response. In this report, intercellular messenger, cytokines, that are regulatory for osteoblast and osteoclast function, were measured. Production of osteocalcin, a marker for osteoblast maturation was also estimated. Human osteoblast-like cells from osteosarcoma cell line MG 63 were grown in wells in the presence of titanium (Ti), titanium alloy (Ti6A14V) and stainless steel implant materials incubated at 37 degrees C. Interleukin-1alpha (IL-1alpha), IL 6, IL-8, IL-11 and osteocalcin were quantitated using standard enzyme linked immunosorbant assay (ELISA) kits from the growth media extracted at specific intervals over the critical ten day period. In all dishes, cells were seen adhering to the base after 24 hours and to confluence at 96 hours. Both IL-1alpha and IL-11 were not produced in sufficient quantities to be measured in the assay (< pg/ml). Interleukin-6 production was significantly higher for stainless steel than for titanium and the alloy. There was a progressive rise in osteocalcin production for titanium contrasted to a basal rate for stainless steel and alloy. Interleukin-8 levels for all metals and controls increased markedly after two days implicating inherent cellular characteristics. A relatively high constant range for macrophage colony stimulating factor from the first day was seen for all metals, including the controls. In conclusion, it appears that titanium implants activate osteocalcin production while stainless steel activates IL-6. PMID- 16312194 TI - The comparative effectiveness of two digit-sucking deterrent methods. AB - A study was conducted to compare the effectiveness of the crib and positive reinforcement in eliminating anterior open bites and increased overjets caused by digit-sucking. The overjet and overbite were measured using an overjet ruler at the start and end of the seventeen-week observation period. Forty patients consented to participate but measurements were only obtained for 11 subjects. The trend in this study is that the crib is more effective than positive reinforcement in preventing digit-sucking. PMID- 16312195 TI - A new surgical management for oro-antral communication: the resorbable guided tissue regeneration membrane--bone substitute sandwich technique. AB - This paper describes a new technique for the closure of oro-antral fistula/communication, in which both hard tissue (bone) and soft tissue closure is achieved. The sandwich technique utilizes a suitable bone grafting material sandwiched between two sheaths of Biogide (a resorbable membrane) for the hard tissue closure of oro-antral communication post traumatic exodontia. The bone grafting material utilized for this case was Bio-oss. The result obtained was excellent with regeneration of sufficient bony tissue to allow placement of an endosseous implant. This sandwich technique is a simple and excellent technique for the closure of oro-antral communication, especially when subsequent placement of endosseous implant is considered without the need of donor site surgery for bone grafting. The otorhinolaryngologists and oral and maxillofacial surgeons should find this technique very useful in the closure of oro-antral fistulae. PMID- 16312196 TI - Cemento-osseous dysplasia in Jamaica: review of six cases. AB - Six cases of cemento-osseous dysplasia (COD) of the jaw bone in Jamaicans are reviewed. Five were documented over a 15-year period (1980-1995). These include a case of florid cemento-osseous dysplasia (previously called gigantiform cementoma). Three of the initial cases were histologically diagnosed as gigantiform cementoma. There was no indication in the patient's case file whether these were familial or non-familial. The other two cases were diagnosed histologically as periapical cemento-osseous dysplasia and cementoblastoma respectively. Based on the current understanding of the nature of florid-cemento osseous dysplasia (FLCOD), a new case was diagnosed as such solely on radiological findings. This single case of FLCOD is reported and discussed against the background of other cemento-osseous lesions. Special emphasis is placed on the radiology of COD in this paper. The confirmative role of radiology without the need for histophathology and treatment for asymptomatic FLCOD is emphasized. PMID- 16312197 TI - Ceftazidime-resistant Salmonella enteritidis in Jamaica. PMID- 16312198 TI - Increased copper level in oral mucosal tissue of patients with submucous fibrosis and who chew areca nut products. PMID- 16312199 TI - Image and diagnosis. Prolapsed nasal polyp. PMID- 16312200 TI - Muscles and lungs: fatal attraction, but time for intervention. PMID- 16312201 TI - Autofluorescence bronchoscopy to identify pre-cancerous bronchial lesions. AB - BACKGROUND: This study aimed to assess the diagnostic yield of autofluorescence bronchoscopy (AFB) in the detection of pre-cancerous bronchial lesions in a non selected sample of patients. METHODS: Both fiberoptic white-light bronchoscopy (WLB) and AFB using the Storz D-light system were performed on 166 consecutive patients. Biopsy specimens were taken in areas of the tracheobronchial tree judged as abnormal or suspicious at WLB and/or AFB. The bronchoscopic procedures were randomly performed by two operators. RESULTS: A total of 93 patients had a positive biopsy specimen: 80 for cancer and 13 for dysplasia. AFB was abnormal or suspicious in 85 of the 93 patients with a sensitivity of 91.4%. Specificity was 50.7%. In 16 patients with normal WLB examination, AFB identified abnormal or suspicious areas which had a positive biopsy. Thus AFB significantly improved sensitivity of WLB (100% vs 82.8%, respectively, p<0.001) in the entire sample of patients studied. Data was further analysed separately for patients with dysplasia and those with cancer. Indeed, 13 of 16 patients recognized only by AFB had a histological diagnosis of dysplasia. The remaining three patients had a diagnosis of cancer (small intraepithelial neoplastic lesions). Since no other patient with dysplasia was found, AFB had a sensitivity of 100% in diagnosing dysplasia. On the other hand, excluding the 13 patients with dysplasia, WLB had a high sensitivity in diagnosing cancer (93.7%). CONCLUSIONS: The AFB Storz system showed a high sensitivity. The increase in diagnostic yield of AFB in comparison with WLB was related to the power of AFB to identify pre-cancerous bronchial lesions so showing its usefulness in the early diagnosis of lung cancer. PMID- 16312202 TI - Different outcomes of pulmonary rehabilitation in patients with COPD with or without exacerbations. AB - BACKGROUND: Pulmonary rehabilitation is recognised as an effective treatment in reducing disability and improving the quality of life in patients with COPD. We evaluated the effects of a course of pulmonary rehabilitation in improving the physical performance and lung function in patients with or without COPD exacerbations. METHODS: 74 patients with COPD were enrolled, 37 (24 males and 13 females, mean age 74.6 years) without exacerbations (group A), and 37 (23 males, 14 females, mean age 73.9 years) with exacerbations (group B). The latter must have had the latest exacerbation at least one month before the inclusion. All patients underwent to a rehabilitation programme of 8 visits in 4 weeks in a day hospital setting, with exercise training, respiratory muscle training and education on COPD. The changes in physical performance and lung function in respect to baseline were measured by a 6-minute walking test, using phethysmography, and by an analogic manometer measuring maximal inspiratory and expiratory pressures (MIP, MEP). RESULTS: Patients of group A showed a mean increase in timed walk distance of 58.38 +/- 57.46 m, compared to a mean increase of 31.38 +/- 44.78 m in group B patients (p = 0.028). As to lung function, a mean increase of 178.92 +/- 132.28 ml in FEV1 in group A versus 67.84 +/- 102.04 ml in group B (p < 0.0001) and a mean increase of 22.36 +/- 25.06 cm H2O in MEP in group A versus 7.70 +/- 12.28 cm H2O in group B (p = 0.002) was found. CONCLUSIONS: These findings indicate that patients with COPD with exacerbations achieve a less favourable outcome of pulmonary rehabilitation, with a significantly lower improvement of physical performance, respiratory muscle strength and lung function in respect to subjects without exacerbations. PMID- 16312203 TI - Alpha-1 antitrypsin deficiency in Italy: regional differences of the PIS and PIZ deficiency alleles. AB - BACKGROUND: Critical to the effective diagnosis and management of disease is information on its prevalence in a particular geographic area such as Italy. Alpha-1 antitrypsin deficiency (AAT Deficiency) is one of the most common serious hereditary diseases in the world, but its prevalence varies markedly from one country to another. AAT Deficiency affects at least 120.5 million carriers and deficient subjects worldwide for the two most prevalent deficiency alleles PIS and PIZ. This genetic disease is known to exist in Italy and is related to a high risk for development of jaundice in infants, liver disease in children and adults, and pulmonary emphysema in adults. METHODS: Studies on the genetic epidemiology of AAT Deficiency has resulted in the development of a unique database that permits a unique analysis of the geographic distribution in 14 different regions located at random from Piemonte to Sicilia. RESULTS: The use of Hardy-Weinberg statistical analysis to evaluate the distribution of these two deficiency alleles has demonstrated striking differences in the frequencies of these two deficiency alleles in these 14 different regions with 23/84 pair wise combinations significantly different (P=0.05) for PIS, and 5/84 combinations for PIZ. CONCLUSIONS: These findings demonstrate differences that impact the standards of care and diagnosis of AAT Deficiency in Italy since the prevalence of these deficiency alleles is not uniform throughout the country. PMID- 16312204 TI - Lung and respiratory muscle function at discharge from a respiratory intensive care unit. AB - BACKGROUND: The purpose of this prospective observational study was to describe lung and respiratory muscle function at Respiratory Intensive Care Unit (RICU) discharge after a severe exacerbation of Chronic Obstructive Pulmonary Disease (COPD). METHODS: The study was conducted in 42 consecutive COPD patients in whom arterial blood gases, dynamic and static lung volumes, maximal inspiratory pressure (MIP) were assessed at discharge from the RICU and compared with values measured 6 months previously when they were in a stable state. The same measurements were performed at 6-month interval in 42 comparable stable COPD patients not requiring any hospitalisation for at least 6 months used as controls. RESULTS: 24% of patients in the study group were discharged with hypercapnia whereas they were normocapnic before the acute episode. Compared to prior to exacerbation, patients of study group showed a significant worsening in mean values of PaCO2 (p=0.005), MIP (p=0.005) and FEV1 (p=0.041). Predefined criteria of worsening in PaCO2, MIP and FEV1 were observed in 47%, 33% and 28% of patients in study respectively. Neither lung nor respiratory muscle function in last stable state did predict post RICU functional worsening. In a period of 6 months controls showed no change in the studied parameters. CONCLUSIONS: After a severe acute exacerbation requiring admission to a RICU and immediately before discharge 1) a large proportion of COPD patients still show preserved lung and respiratory muscle function 2) more than one third of them would require further care and rehabilitative attempts to restore functional derangements. PMID- 16312205 TI - An overview on severe acute respiratory syndrome (SARS). AB - Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease that has caught the medical profession by surprise in 2003. The major clinical features include persistent fever, chills/rigor, myalgia, malaise, dry cough, headache and dyspnoea but diarrhea occurs in 40-70% of patients after hospital admission. Respiratory failure is the major complication of SARS; at least half of the patients require supplemental oxygen during the acute phase whereas about 20% of patients progress to acute respiratory distress syndrome requiring invasive mechanical ventilatory support. In contrast, the severity is generally mild in infected young children. Due to our limited understanding of this new disease, treatment of SARS was empirical in 2003. Protease inhibitor (Lopinavir/ritonavir) in combination with ribavirin may play a role as antiviral therapy in the early phase whereas nelfinavir is a promising alternative. The role of interferon and systemic steroid in preventing immune-mediated lung injury deserves further investigation. In addition, other anti-viral treatment, RNA interference, monoclonal antibody, synthetic peptides, and vaccines are being developed. Rapid diagnosis, early isolation, and good infection control measures are important in preventing spread of the infection. PMID- 16312206 TI - Comparison of tuberculin skin test and Quantiferon immunological assay for latent tuberculosis infection. AB - BACKGROUND: Correct identification of individuals with latent tuberculosis infection (LTBI) is a crucial element of the elimination strategy, allowing their adequate treatment. In addition to tuberculin skin test (TST), the Quantiferon test (QFT, based on whole blood gamma-interferon release) had been recently proposed. Aim of the study is to compare this test to TST for identification of LTBI in a non-selected population, in order to verify their value in identifying truly infected individuals (entitled to receive preventive chemotherapy), and to exclude from treatment those having a positive TST for other reasons (e.g. after BCG vaccination). METHODS: 136 consecutive persons (78 males, mean age 34 +/- 9 years) referred to the clinic for TST were recruited (78 born in low--or middle- income countries). Based on their history, the cases were divided into 4 groups: 1) recently traced contacts of whom 18 TST negative and 28 TST positive; 2) 22 screening subjects, all TST negative; 3) BCG vaccinated subjects (14); and 4) 54 subjects already undergoing treatment of LTBI for exposure to TB. RESULTS: The overall agreement between TST and QFT was 72% (64% in TST positive and 88.4% in TST negative subjects). The proportion of TST positive/QFT negative BCG vaccinated individuals was 23.1%. The K coefficient was 0.474 in recently traced contacts, 0.366 in BCG vaccinated individuals and 0.451 overall. CONCLUSIONS: The study results suggest that agreement between TST and QFT is lower in TST positive than in negative subjects, being lower in individuals treated for LTBI. Quantiferon does not seem to have brought significant improvement in the diagnosis of LTBI. PMID- 16312207 TI - Successful management of ARDS with bronchopleural fistula secondary to miliary tuberculosis using a conventional ventilator. AB - Most institutions in India and other developing countries do not have facilities for high frequency ventilation in adults. We report the successful management of a case of ARDS with bronchopleural fistula secondary to miliary tuberculosis using a conventional ventilator and early empiric anti-tubercular therapy. PMID- 16312208 TI - Extrapleural hematoma: when is surgery indicated? AB - Extrapleural haematoma (EH) is a rare clinical condition of which four cases are presented. Different etiologies were encountered. Identical findings were found on the chest X-ray. Two cases were secondary to spontaneous rib fracture and the other two were iatrogenic. Surgical exploration was necessary in one patient following pleural biopsy. Medical treatment and clinical observation were the treatment of choice in the remaining three stable patients. PMID- 16312209 TI - Liposarcoma of the pleural cavity. An exceptional tumour. AB - Primitive liposarcomas of the pleura are exceptional tumours. We report a new case of primitive liposarcoma of the pleura revealed by chest pains in a 50 year old man. Computed tomography showed a large fat density mass in the left pleural cavity. Surgical resection was performed, completed with adjuvant radiotherapy. Few reports are available in the literary world. We present our case, review previously reported cases and discuss treatment. PMID- 16312210 TI - Primary pulmonary nodular amyloidosis. AB - Primary nodular amyloidosis of the lung is an uncommon manifestation. The disease runs a benign course, but offers diagnostic problems due to non-specific radiological features entering the big field of the solitary nodule. We describe the case of a 60 year old man with multiple nodules on the left lung operated on diagnostic and therapeutic video-assisted thoracoscopy and discuss the possibilities, if any, of suspecting such a disease through radiologic characteristics along with findings from the patient's history, physical examination and laboratory tests. PMID- 16312211 TI - Single-cell imaging: shedding new light. PMID- 16312212 TI - Modification of LongSAGE for obtaining and cloning long concatemers. PMID- 16312213 TI - Unlabeled oligonucleotide probes modified with locked nucleic acids for improved mismatch discrimination in genotyping by melting analysis. PMID- 16312214 TI - Clearing and photography of whole mount X-gal stained mouse embryos. PMID- 16312215 TI - Isolation of vaccinia MVA recombinants using the viral F13L gene as the selective marker. AB - Modified vaccinia Ankara (MVA) is a highly attenuated vaccine vector that has an excellent vaccine safety record. Also, as a eukaryotic gene expression vector, MVA can be used in a biosafety level 1 setup, in contrast to more virulent vaccinia virus strains. Isolation of recombinant MVA involves repeated plaquing of the virus and is burdensome because virus plaques are slow to develop and difficult to recognize. To facilitate the generation of MVA recombinants, we have developed a cloning system for MVA based on the selection of the viral F13L gene. Deletion of F13L in MVA produced a small plaque phenotype and a reduction in extracellular virus formation, indicating a severe block in cell-to-cell spread. When using the F13L knockout virus as the parental virus, reintroduction of the F13L gene in the original locus was used as an efficient selection for the isolation of virus recombinants. The selection procedure can be done entirely in the permissive baby hamster kidney (BHK)-21 cell line, does not require plaque isolation, and rendered close to 100% recombinant virus. PMID- 16312216 TI - insilico.mutagenesis: a primer selection tool designed for sequence scanning applications used in directed evolution experiments. AB - Several primer prediction programs have been developed for a variety of applications. However none of these tools allows the prediction of a large set of primers for whole gene site-directed mutagenesis experiments using the megaprimer method. We report a novel primer prediction tool (insilico.mutagenesis), accessible at www.insilico.uni-duesseldorf.de, developed for the application to high-throughput mutagenesis used in directed evolution or structure-function dependency projects, which involve the subsequent mutagenesis of a large number of amino acid positions (e.g., in whole gene saturation or gene scanning mutagenesis experiments). Furthermore, the program is suitable for all site directed (saturation) mutagenesis approaches, such as saturation mutagenesis of promoter sequences and other types of untranslated intergenic regions. In anticipation of downstream cloning steps, the primer design tool also includes a restriction site control feature alerting the user if unwanted restriction sites have been introduced within the mutagenesis primer. The use of our tool promises to speed up the process of site-directed mutagenesis, as it instantly allows predicting a large set of primers. PMID- 16312217 TI - High-throughput aggregate culture system to assess the chondrogenic potential of mesenchymal stem cells. AB - We have developed an improved method for preparing cell aggregates for in vitro chondrogenesis studies. This method is a modification of a previously developed conical tube-based culture system that replaces the original 15-mL polypropylene tubes with 96-well plates. These modifications allow a high-throughput approach to chondrogenic cultures, which reduces both the cost and time to produce chondrogenic aggregates, with no detrimental effects on the histological and histochemical qualities of the aggregates. We prepared aggregates in both systems with human bone marrow-derived mesenchymal stem cells (hMSC). The aggregates were harvested after 2 and 3 weeks in chondrogenic culture and analyzed for their ability to differentiate along the chondrogenic pathway in a defined in vitro environment. Chondrogenic differentiation was assessed biochemically by DNA and glycosaminoglycan (GAG) quantification assays and by histological and immunohistologic assessment. The chondrogenic cultures produced in the 96-well plates appear to be slightly larger in size and contain more DNA and GAG than the aggregates made in tubes. When analyzed histologically, both systems demonstrate morphological characteristics that are consistent with chondrogenic differentiation and cartilaginous extracellular matrix production. PMID- 16312218 TI - Concentration and isolation of DNA from biological fluids by agarose gel isotachophoresis. AB - Isotachophoresis is an electrophoretic method of separation of charged substances. The method is characterized by a discontinuous buffer system, constant velocity of separated molecules, and the distribution of separated components in the form of narrow concentrated bands located one right after another. As a rule, isotachophoresis is not used for the separation of nucleic acids because the mobility of polynucleotides in this system does not depend on their size. However, this circumstance proved to be very useful for the quantitative isolation of heterogeneous DNA fragments from biological fluids, for gene diagnostics of cancer in particular. The proposed method of agarose gel isotachophoresis of DNA has been used for the isolation of blood DNA and its successful PCR analysis. PMID- 16312219 TI - Four-color, 4-D time-lapse confocal imaging of chick embryos. AB - Our understanding of the molecular mechanisms that direct cell motility, cell division, and cell shaping has benefited from innovations in cell labeling and the ability to resolve intracellular dynamics with multispectral, high-resolution imaging. However, due to difficulties with in vivo cell marking and monitoring, most studies have been restricted to fixed tissue or cells in culture. Here, we report the delivery of multiple (up to four), multicolor fluorescent protein (FP) constructs and four-dimensional (4-D), multispectral time-lapse confocal imaging of cell movements in living chick embryos. Cell cytoskeletal components are fluorescently tagged after microinjection and electroporation of a cocktail of FP constructs into specific regions of chick embryos. We tested 11 different FP constructs in various two-, three-, and four-color combinations using multispectral imaging and linear unmixing to limit the crosstalk between different emission spectra. We monitored intracellular dynamics in individual multicolored migrating cells in vivo and developed a set of advantageous imaging parameters for 4-D time-lapse confocal microscopy. We find that the number of four-color labeled cells in a typical embryo is approximately 10% of the total number of fluorescently labeled cells; this value consistently increases showing that approximately 50% of the total labeled cells have only one-color. We find that multicolored cells are photostable for time-lapses of approximately 2-3 h. Thus, cell labeling with up to four FP color schemes combined with multispectral, 4-D confocal time-lapse imaging offers a powerful tool to simultaneously analyze cellular and molecular dynamics during chick embryogenesis. PMID- 16312220 TI - Method for discovering novel DNA viruses in blood using viral particle selection and shotgun sequencing. AB - Rapid identification of viruses is needed to monitor the blood supply for emerging threats. Here we present a method that meets these criteria and allows for the shotgun sequencing of novel, uncultured DNA viruses directly from human blood. This method employs selection based on the physical properties of viruses combined with sequence-independent amplification and cloning. We show that both single- and double-stranded DNA viruses can be recovered from blood samples using this approach. In addition, we report the discovery of novel anellovirus sequences in the blood of healthy donors. PCR primers designed to amplify these novel anellovirus sequences were then used to verify the presence of these viruses in the general donor population. PMID- 16312221 TI - Infectious pathogen detection arrays: viral detection in cell lines and postmortem brain tissue. AB - A unique array-based pathogen chip has been developed for the detection of viral RNA or DNA relevant to pathologies of the central nervous system. A total of 715 unique oligonucleotides (60-mer) representing approximately 100 pathogens were designed based on open reading frames (ORFs) from highly conserved and heterogenic regions within viral families. In addition, viral genes reflecting different stages of pathogen infection were also included to potentially define the stage of the viral infection. Viruses (double-stranded DNA, double- or single stranded RNA, delta, retroid), parasites, and bacteria were included. Test samples labeled with Cy5 were examined by cohybridization with a reference RNA, labeled with Cy3, to the pathogen microarray chip. Good reproducibility of experiments was observed, based on data generated from duplicate hybridizations and duplicate spots on the microarray platform. A viral transcript detection sensitivity of 1 x 10(3) plaque-forming units (pfus) was achieved using selected cell lines and viruses. These findings suggest that the array-based platform described here is capable of detecting a broad spectrum of viruses in a single assay with relatively high sensitivity, specificity, and reproducibility. This method may be used to provide evidence of viral infection in postmortem tissue from psychiatric patients as well as a wide range of other diagnostic categories. PMID- 16312222 TI - Evaluation of arrayed primer extension for TP53 mutation detection in breast and ovarian carcinomas. AB - Mutations in the tumor suppressor gene TP53 are associated with a wide range of different cancers and may have prognostic and therapeutic implications. Methods for rapid and sensitive detection of mutations in this gene are therefore required. In order to make screening more effective, a commercially available TP53 genotyping microarray from Asper Biotech has been constructed by arrayed primer extension (APEX). The present study is the first report that blindly evaluates the efficiency of the second generation APEX TP53 genotype chip outside the Asper laboratory and compares it to temporal temperature gradient electrophoresis (TTGE) and sequencing of TP53 for mutation detection in ovarian and breast cancer samples. All nucleotides in the TP53 gene from exon 2-9 are included on the chip by synthesis and application of sequence-specific oligonucleotides. The chip was validated by screening 48 breast and 11 ovarian cancer cases, all of which had previously been analyzed by TTGE and sequencing. APEX scored 17 of 20 sequence variants, missing one deletion, one insertion, and a missense mutation. Resequencing efficiency using APEX was 92% for both DNA strands and 99.5% for sense and/or antisense strand. We conclude that the APEX TP53 microarray is a robust, rapid, and comprehensive screening tool for sequence alterations in tumors. PMID- 16312223 TI - From one generation to the next: a legacy of holistic nursing education. PMID- 16312224 TI - One nurse shares her quest for knowledge. PMID- 16312225 TI - Caring cultures. PMID- 16312226 TI - Some tips for choosing programs on alternative modalities. PMID- 16312227 TI - Evidence-based practice in the holistic nursing process. PMID- 16312228 TI - New educational opportunities to improve competencies in geriatric care. PMID- 16312229 TI - Ten certifiable steps down the road to quality care. PMID- 16312230 TI - Canine cutaneous clear cell adnexal carcinoma: histopathology, immunohistochemistry, and biologic behavior of 26 cases. AB - Thirty tumors including 27 distinctive cutaneous neoplasms and 3 metastatic tumors from 26 dogs were collected from diagnostic submissions to 3 laboratories. Characteristic histopathologic features included location in the subcutis or dermis (or both); lobular, nodular, and nest-like architecture; and a component of epithelioid cells with clear cytoplasm. Additional features present in most cases included follicular dermal papilla-like structures, low mitotic index, nuclear pleomorphism, necrosis, and mineralization. Cytoplasmic periodic acid Schiff-positivity, which was abolished by pretreatment with diastase, indicated the presence of glycogen in all cases. The oil red O stain did not demonstrate cytoplasmic lipid. Melanin granules, accentuated by the Fontana-Masson method, were observed infrequently. A sparsely cellular mucinous stroma and stromal cartilaginous differentiation were uncommon. By immunohistochemistry, neoplastic cells stained positively for cytokeratin (29 of 29), vimentin (28 of 28), S-100 protein (24 of 29), and melan A (8 of 12); results were negative for smooth muscle actin and calponin in all cases. Clinical follow-up information was obtained on all 26 dogs. One tumor recurred, 1 metastasized to a regional lymph node, and 1 metastasized to regional lymph nodes twice. In another case, possible pulmonary metastasis was noted radiographically. The findings are consistent with a poorly differentiated, low-grade, adnexal carcinoma of the skin. Similar canine cutaneous neoplasms have been reported as "clear-cell hidradenocarcinoma" and "follicular stem cell carcinoma." The authors propose the designation "cutaneous clear cell adnexal carcinoma." PMID- 16312231 TI - Development of an enzyme-linked immunosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood. AB - Equine inflammatory disease is accompanied by a neutrophil activation resulting in the release of granulocytic enzyme myeloperoxidase (MPO). To measure MPO in horse plasma as marker of neutrophil activation, the authors purified equine neutrophil MPO and developed a specific enzyme immunoassay using 2 specific polyclonal antibodies obtained from rabbit (primary antibody) and guinea pig (secondary antibody). The sandwich complex "primary antibody-MPO-secondary antibody" was detected using a goat anti-guinea pig immunoglobulin antibody conjugated to alkaline phosphatase. The enzyme-linked immunosorbent assay (ELISA) showed good precision and accuracy, with intra- and interassay coefficients of variation below 10% for MPO concentrations ranging from 0.78 to 50 ng/ml. A stable plasma MPO value, unaffected by time elapsed between blood collection and centrifugation, was obtained with plasma from EDTA anticoagulated blood. The mean MPO value measured in 38 healthy horses was 181.80 +/- 64.74 ng/ml. In 20 horses suffering from obstruction of the large or small intestine, MPO concentrations measured at the time of arrival at the intensive care unit were significantly higher than mean normal value, ranging from 477.88 to 2,748.13 ng/ml. Work is in progress to apply this MPO ELISA technique to other biological fluids and other equine diseases. PMID- 16312232 TI - Genetic analysis of feline caliciviruses associated with a hemorrhagic-like disease. AB - Feline calicivirus (FCV) is 1 of the most common causes of upper respiratory tract disease in cats. Other disease syndromes associated with FCV infection have been reported. Recently, calicivirus infection associated with a hemorrhagic-like disease leading to significant mortality in cats has been reported. The clinical signs are similar to those observed with the calicivirus of rabbit hemorrhagic disease. This study characterized 2 FCV isolates associated with hemorrhagic-like disease. Nucleotide sequencing of the complete genome has been done for these 2 isolates as well as for 4 additional isolates representing other disease syndromes. Previously reported sequence data for the entire genome of classical FCV (6 isolates) and a portion of the capsid gene for hemorrhagic-like FCV (3 isolates), isolated in different regions of United States were used in the genetic analysis. Sequence data were used to determine relationships among the isolates and any correlation with phenotype. Nucleotide sequence comparisons of the entire genome and individual open reading frames revealed high homology among all isolates. Data suggest that the virulence may have genetic determinants on the basis of phylogenetic clustering of the isolates associated with hemorrhagic like disease. PMID- 16312233 TI - The adequacy of sample type/weight and incubation period on detection of Salmonella spp. in slaughter cattle. AB - The purpose of this study was to examine the adequacy of different sample types (fecal and rumen content), rumen-content sample weight (1, 10, and 25 g), and incubation period on the detection of Salmonella spp. in grass-fed beef cattle at slaughter. The culture technique was the same for all samples and followed the Australian Standard (AS 1766.2.5-1991). Sample adequacy was defined as the ratio between the overall prevalence, as obtained from samples identified as positive by any sample type/weight, and the estimated prevalence, as obtained from samples identified as positive by a particular sample type/weight. Sample adequacy reflects the likelihood of a sample of a particular type and weight to contain the organism of interest and hence is related to the sensitivity of the diagnostic test. It was found that sample adequacy differed between sample types and weights: 37.5% for both a 10-g fecal sample and a 1-g rumen sample, 77.1% for a 10-g rumen sample, and 79.2% for a 25-g rumen sample. On this basis, it is strongly recommended that sample type and weight be considered in the design of studies that aim to quantify Salmonella contamination in cattle. PMID- 16312234 TI - In vitro antimicrobial inhibition profiles of Mycoplasma bovis isolates recovered from various regions of the United States from 2002 to 2003. AB - Antimicrobial therapy continues to be important in reducing losses due to pneumonic forms of Mycoplasma bovis disease in beef and dairy calves. Although M. bovis diseases have been documented as frequent and economically important in the United States, there are no published reports on the antimicrobial activity of approved compounds against US strains. In this study, the authors report on the activity of 9 different antimicrobials against 223 recently recovered isolates of M. bovis. These isolates represent accessions from 5 geographic regions of the United States and were grouped by 4 tissues of origin (milk, respiratory, joint, or ear and eye). A broth microdilution test was used to determine minimum inhibitory concentration (MIC) values by reading redox changes detected in broth with alamarBlue (resazurin) indicator. For each antimicrobial, the median, MIC50, MIC90, mode, and range were calculated, and the values used for comparisons. In the absence of accepted breakpoint values, published MIC cutoff values for animal mycoplasmas as well as Clinical Laboratory Standards Institute interpretive criteria were used as a reference to define in vitro activity. The MIC values from active antimicrobials were found to distribute independently of region of origin of the isolates or of tissue of origin. Enrofloxacin, florfenicol, and spectinomycin were found to be active compounds in vitro. Oxytetracycline and chlortetracycline were active against more than half of the isolates. Very few isolates were inhibited by tilmicosin and none by erythromycin, ampicillin, or ceftiofur. The antimicrobial profiles determined for these US strains were remarkably similar to those reported for European isolates. However, unlike in Europe, there appears to be no diversity of profiles when US isolates are grouped by region or tissue of origin. PMID- 16312235 TI - Use of avidity enzyme-linked immunosorbent assay and avidity Western blot to discriminate between acute and chronic Neospora caninum infection in cattle. AB - Avidity serological tests (avidity enzyme-linked immunosorbent assay [ELISA] and avidity Western blot) were developed and used to differentiate between acute (primary infection, reinfection, and recrudescence) and chronic Neospora caninum infection in cattle. In addition, the pattern of immunoglobulin G (IgG) avidity maturation against different specific antigens of N. caninum tachyzoites was studied. Sequential serum samples were collected from cattle naturally and experimentally infected with N. caninum. Four groups of experimentally infected cattle were included in the study and were representative of primary infection, reinfection, chronic infection, and noninfection. Serum samples were also collected from naturally infected cattle classified into nonaborting and aborting cows on the basis of clinical findings and serological profiles, and a third group composed of seronegative cows that seroconverted during the course of the experiment. All samples were tested by avidity ELISA and avidity Western blot. The IgG avidity ELISA allowed the discrimination between primary and chronic infection because all experimentally primary-infection cows showed low avidity indexes at week 4 postinfection (p.i.) compared with the high avidity values found at week 20 postinfection. However, this test did not allow the discrimination of reinfection or recrudescence from chronic infection. Regarding IgG avidity Western blot results, no antigenic markers correlating with acute (primary infection, recrudescence, and reinfection) or chronic infection were recognized. However, the 17-kD immunodominant antigen was mostly responsible for high avidity values obtained by avidity ELISA because it was intensively recognized by high-avidity antibodies in all chronically infected animals after urea treatment. PMID- 16312236 TI - Evaluation of a new antibody-based enzyme-linked immunosorbent assay for the detection of bovine leukemia virus infection in dairy cattle. AB - The objective of this study was to validate a new blocking enzyme-linked immunosorbent assay (ELISA) (designated M108 for milk and S108 for serum samples) for detecting bovine leukemia virus (BLV) infection in dairy cattle. Milk, serum, and ethylenediaminetetraacetic acid-blood samples were collected from 524 adult Holstein cows originating from 6 dairy herds in Central Argentina. The M108 and S108 were compared with agar gel immunodiffusion (AGID), polymerase chain reaction and a commercial ELISA. Because there is currently no reference test capable of serving as a gold standard, the test sensitivity (SE) and specificity (SP) were evaluated by the use of a latent class model. Statistical inference was performed by classical maximum likelihood and by Bayesian techniques. The maximum likelihood analysis was performed assuming conditional independence of tests, whereas the Bayesian approach allowed for conditional dependence. No clear conclusion could be drawn about conditional dependence of tests. Results with maximum likelihood (under conditional independence) and posterior Bayes (under conditional dependence) were practically the same. Conservative estimates of SE and SP (with 95% confidence intervals) for M108 were 98.6 (96.7; 99.6) and 96.7 (92.9; 98.8) and for S108 99.5 (98.2; 99.9) and 95.4 (90.9; 98.1), respectively. The ELISA 108 using either milk or serum to detect BLV-infected animals had comparable SE and SP with the official AGID and a commercial ELISA test, which are currently the most widely accepted tests for the serological diagnosis of BLV infection. Therefore, ELISA 108 can be used as an alternative test in monitoring and control programs. PMID- 16312237 TI - Persistence of Neospora caninum-specific immunoglobulin G antibodies in bovine blood and lung tissue stored at room temperature. AB - Blood and tissue samples obtained at necropsy from carcasses showing varying degrees of postmortem decomposition are sometimes used as sources of antibodies for serological tests, primarily in studies involving wild animals or in examination of aborted fetuses. To test the persistence of antibodies in blood and lung tissue kept at room temperature (approximately 20degreesC) for up to 48 and 118 days, respectively, serum and lung fluid from Neospora caninum-infected and uninfected cattle were tested using an indirect N. caninum immunostimulating complex enzyme-linked immunosorbent assay (ELISA). Testing of selected samples was also performed using a commercial Neospora competitive ELISA. Despite severe changes in the gross appearance of the samples, the blood collected through venipuncture from live animals remained essentially sterile, whereas the pieces of lung tissue showed a variety of bacterial and fungal growth. Nevertheless, there was no obvious reduction in antibody levels in either the blood or the tissue samples during the 2 observation periods. This study indicates that specific immunoglobulin G antibodies may be remarkably stable in sterile blood and decomposed lung tissue. PMID- 16312238 TI - Effects of delayed or prolonged fixation on immunohistochemical detection of bovine viral diarrhea virus type I in skin of two persistently infected calves. AB - The effects of delayed or prolonged fixation on immunohistochemical detection of bovine viral diarrhea virus (BVDV) antigen were evaluated in skin. Ear-notch specimens from 2 calves persistently infected with BVDV type 1 were handled in 1 of 3 ways: 1) fixed in formalin promptly and processed for immunohistochemistry (IHC) after 3-176 days; 2) held at 3-4degreesC in plastic bags up to 10 days, then fixed in formalin for 2-5 days before processing; or 3) exposed to room air and temperature for 1-5 days before formalin fixation. Immunohistochemical staining intensity was evaluated without the knowledge of specimen handling. Staining of specimens that had been promptly fixed in formalin was moderate to strong at all fixation periods through 36 days, weak or no staining was evident in specimens fixed for 176 days. Refrigerated specimens typically had moderate to strong immunohistochemical staining. Even after 10 days of refrigeration before fixation, all immunohistochemical reactions were positive. However, no immunohistochemical staining was detected in any specimen that was exposed to room air. Results indicate that prompt formalin fixation is optimal for BVDV IHC. Samples can be held in formalin at least 36 days, without loss of reactivity. A 1 day delay in fixation caused no loss of reactivity, provided the specimen was refrigerated and protected from desiccation. PMID- 16312239 TI - Comparison of two enzyme-linked immunosorbent assays for diagnosis of Mycobacterium avium subsp. paratuberculosis. AB - Enzyme-linked immunosorbent assays (ELISAs) are used in Johne's disease (JD) control programs as a first screening for presence of the disease in a herd. A high sensitivity of the ELISA is therefore important, yet the commonly used ELISAs have relatively low sensitivity. The inclusion of an absorption phase, although improving specificity, potentially decreases sensitivity. Sera and feces of 383 adult dairy cows in 8 herds were used to compare the test characteristics of an absorbed and a nonabsorbed indirect ELISA for the detection of JD. The absorbed ELISA is based on a protoplasmic antigen, whereas the nonabsorbed uses a lipoarabinomannan-based antigen. The potential advantage of the nonabsorbed ELISA is that it may be less specific and more sensitive. Two herds certified free of JD were used to compare the specificity of the ELISAs. The other herds used to compare sensitivity were either infected with Mycobacterium avium subsp. paratuberculosis or had unknown status. Using fecal culture as a gold standard, the diagnostic specificity for the absorbed and nonabsorbed ELISAs were 98.4% and 87.9%, respectively. The diagnostic sensitivity was 72.4% and 65.5% for the absorbed and the nonabsorbed ELISA, respectively. Furthermore, a comparison using a fecal DNA probe as the comparison standard resulted in both ELISAs having a sensitivity of 61.9%. Agreement between the 2 ELISAs was moderate, with a kappa statistic of 0.58. The nonabsorbed ELISA did not have a higher sensitivity and had a lower specificity than the absorbed ELISA. Therefore, in this population, there was no advantage gained with using the nonabsorbed ELISA. PMID- 16312240 TI - Comparison of immunohistochemistry and two rapid tests for detection of abnormal prion protein in different brain regions of sheep with typical scrapie. AB - One of the "gold standard" techniques for postmortem confirmation of scrapie diagnosis in sheep and goats is immunohistochemical examination of brain tissue. Active surveillance for scrapie is mainly performed by rapid diagnostic tests on the basis of postmortem immunochemical detection of prion protein (PrP) in the obex tissue. The aim of this study was to determine the performance of 2 rapid tests, Prionics-Check LIA (a chemiluminescence sandwich enzyme-linked immunosorbent assay) and Prionics-Check Western blot for scrapie diagnosis when applied to brain areas other than the obex, in comparison with the recognized immunohistochemistry. Prion protein was detected in the obex, cervical spinal cord, and thalamus from all the scrapie-positive sheep by the 3 tests. Western blot and LIA were negative in other areas of the brain, although weak immunohistochemical staining was detected. The results show that the 2 rapid tests studied may detect PrP in brain areas other than the obex, although with a lower sensitivity than immunohistochemistry when there is minimal PrP deposition. PMID- 16312241 TI - A simple and rapid chromatographic strip test for detection of antibody to porcine reproductive and respiratory syndrome virus. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be a major economic problem for swine industries worldwide despite several disease reduction strategies such as age-segregated early weaning and all-in-all-out pig movement. Routine diagnosis of PRRSV is carried out by the combined use of an antibody-detecting enzyme-linked immunosorbent assay (ELISA), immunofluorescence, reverse transcription-polymerase chain reaction, and virus isolation. These assays require specialized laboratory equipment in addition to multistep sample handling and sample preparation. The objective of this study was to evaluate a simple pen-side assay (BioSign PRRSV) for rapid detection of PRRSV antibody based on a lateral flow chromatographic strip immunoassay system. This assay uses Escherichia coli-expressed viral nucleocapsid protein antigen for detecting antibodies against PRRSV in swine sera. In this report, the authors describe the evaluation of this assay using sera from both clinical samples and experimentally infected piglets. The results were compared with those of a standard, commercially available antibody ELISA (HerdChek PRRS ELISA) and an indirect immunofluorescence assay using the same serum samples. The BioSign PRRSV assay was capable of detecting antibodies in sera known to contain antibodies to PRRSV, resulting in 93.2% sensitivity for samples from experimentally infected pigs and 98.7% sensitivity for clinical serum samples. For sera that did not contain antibodies to PRRSV, the specificity was found to be 98.5% and 99.2% for clinical and experimental serum samples, respectively. PMID- 16312242 TI - Lymphoid follicular cloacal inflammation associated with a novel herpesvirus in juvenile alligators (Alligator mississippiensis). AB - Multifocal hyperemic nodules and plaques associated with the cloacal mucosa of juvenile alligators (Alligator mississippiensis) at a public aquarium were investigated. Grossly, pale pink to dark red multifocal, circular lesions of varying degrees of severity were identified on the cloacal and, in males, phallus mucosa. Cloacal mucosa biopsies were obtained from 2 of the alligators. These samples were examined histologically and by polymerase chain reaction (PCR) using consensus primers targeting a conserved region of the herpesvirus polymerase gene. Microscopically, the lesions were characterized as submucosal lymphoid follicles with hyperemia and hemorrhage. No inclusion bodies were observed. Minimal to no anisokaryosis was present, and no etiologic agents were identified. Through PCR, a band consistent in size with herpesvirus was observed. Tissues showing similar clinical, histopathologic, and PCR findings were collected from animals at an alligator farm several months later. Sequencing of the PCR amplicon resulted in a 180-base pair sequence that shared 85% sequence identity with tortoise herpesvirus-1. PMID- 16312243 TI - Serotyping of US isolates of Chlamydophila psittaci from domestic and wild birds. AB - The identities of chlamydial strains, which can infect a given host, are important to know for disease prognosis, disease control, and epidemiology. The microimmunofluorescence test (MIFT) was used with a panel of 14 serovar-specific monoclonal antibodies (MAbs) to serotype 150 chlamydial isolates from domestic and wild birds. The isolates were obtained from birds submitted to diagnostic laboratories or during investigation of outbreaks. The 150 US isolates included 96 from the order Psittaciformes, 14 isolates from the order Columbiformes, 2 from the order Passeriformes, 16 from the order Galliformes, 12 from the order Struthioniformes, and 3 from the order Falconiformes. A total of 93, or 97%, of the Psittaciformes isolates were of serovar A; 11, or 79%, of the Columbiformes isolates were of serovar B; 64% of the Galliformes isolates were of serovar D, and all the Struthioniformes isolates were of serovar E. The 3 Falconiformes isolates did not react with any of the MAbs to the avian and mammalian isolates and are presumed to represent a new strain. The results show that specific chlamydial strains are usually associated with certain types of birds and that some serovars may be unusually virulent for certain species of birds. The MIFT using serovar-specific MAbs provides a rapid method to serotype new isolates, making it a useful system for epidemiological studies. PMID- 16312244 TI - Investigation of falsely reported resistance of Streptococcus equi subsp. zooepidemicus isolates from horses to trimethoprim-sulfamethoxazole. AB - The objective of this study was to investigate the perceived increase in resistance of Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) isolated from the lower respiratory tract of horses to trimethoprim-sulfamethoxazole (SXT). The recorded SXT-susceptibility results of 50 S. zooepidemicus isolates from the tracheal wash fluid of equine patients examined at Colorado State University Veterinary Teaching Hospital from each of 2 time periods (1987-1990 and 1997-2001) were compared and statistically analyzed using a cross-sectional study design. There was a statistically significant difference between the documented resistance of S. zooepidemicus isolated in the 1987-1990 time period (8%), using quantitative microbroth dilution, and the resistance reported for isolates from the 1997-2001 time period (42%), using Kirby-Bauer agar disk diffusion. Laboratory investigation revealed inadequate quality control of media and subsequent falsely reported resistance of S. zooepidemicus from 1997 to 2001 time period. This study demonstrates how minor deviations from prescribed laboratory-testing guidelines can have a major effect on antimicrobial susceptibility test results. The study also underscores the need for regular surveillance and monitoring of trends in antimicrobial susceptibility to detect and correct such problems. In addition, epidemiologists and others collecting data from laboratories should be cautioned to interact with the laboratory regarding interpretation of results of various testing methods to ensure accurate analysis and conclusions. PMID- 16312245 TI - Distribution of Crenosoma vulpis and Eucoleus aerophilus in the lung of free ranging red foxes (Vulpes vulpes). AB - Crenosoma vulpis and Eucoleus aerophilus are nematode parasites that can cause verminous pneumonia in wild carnivores. There is a paucity of information regarding the distribution of parasites in the lungs and the relationship between histopathological and parasitological diagnoses in naturally infected foxes. The objectives of this study were: first, to study the lobar and airway distribution of C. vulpis and E. aerophilus in wild red foxes and second, to investigate the relationship between fecal and histopathological diagnoses. Samples from 6 sites of the lung and fecal contents were obtained from 51 wild foxes in Prince Edward Island. By fecal examination, 78.4% of wild foxes tested positive for C. vulpis and 68.6% for E. aerophilus. In contrast, 66.6% and 49% of foxes had histopathological evidence of C. vulpis and E. aerophilus in the lungs, respectively. Anatomically, C. vulpis was observed in the small bronchi and bronchioles of all pulmonary lobes whereas E. aerophilus was restricted to the large bronchi and the caudal lobes. Affected airways exhibited severe epithelial glandular hyperplasia and bronchiolar mucous metaplasia. It was concluded that C. vulpis is widely distributed in airways of all pulmonary lobes, whereas E. aerophilus is mainly restricted to the bronchi of caudal lobes. Also, this study showed that histological examination of lung underestimates the infection with E. aerophilus. PMID- 16312246 TI - Segmental aplasia of uterine body in an adult mixed breed dog. AB - Segmental aplasia of the uterine body was diagnosed in a 5-year-old, mixed breed bitch. Abdominal radiography and transabdominal ultrasonography revealed marked dilation of fluid-filled uterine horns with no evidence of a uterine body. Sex hormone assays did not detect the presence of estradiol-17 beta; however, progesterone (2 ng/ml) was found in the serum, indicating anestrus. On gross examination of the reproductive tract, the uterine body was absent, apparently never formed. In its place, a cord-like piece of tissue was identified as an aplastic/dysplastic remnant, connecting the cervix and right uterine horn. The tip of the cord-like piece branched into 5 string-like pieces of tissue, 1 of which was connected to the region dividing the left and right uterine horns. Both the uterine horns were dilated markedly revealing hydrometra. Histologically, uterine body remnant tissues from the endometrium, myometrium, and perimetrium were detected in proximal and distal parts of the uterine body. The string-like piece of tissue connecting the uterine body remnant and the uterine horn consisted of a round cluster of smooth muscle cells surrounding a central core of adipose tissue with blood vessels. It was concluded that the hydrometra observed in both uterine horns was induced by an obstruction resulting from segmental aplasia in the uterine body. This is the first known report of segmental aplasia in the uterine body of a bitch. PMID- 16312247 TI - Spontaneous aortic dissecting hematoma in two dogs. AB - This report describes 2 cases of spontaneous aortic dissecting hematoma in young Border Collie and Border Collie crossbred dogs. Histology was performed in one of the cases involving an unusual splitting of the elastin present within the wall of the aorta, consistent with elastin dysplasia as described in Marfan syndrome in humans. The first case involved a young purebred Border Collie that died suddenly and the second case involved a Border Collie crossbred dog that died after a 1-month history of seizures. Gross lesions included pericardial tamponade with dissection of the ascending aorta in the former case and thoracic cavity hemorrhage, mediastinal hematoma, and aortic dissection in the latter. Histologic lesions in the case of the Border Collie crossbred dog included a dissecting hematoma of the ascending aorta with elastin dysplasia and right axillary arterial intimal proliferation. PMID- 16312248 TI - Erysipelothrix rhusiopathiae septicemia in a Laughing kookaburra (Dacelo novaeguineae). AB - Erysipelothrix rhusiopathiae (E. rhusiopathiae) septicemia was demonstrated in a captive Laughing kookaburra (Dacelo novaeguineae). The bird died after a 2-week period of weakness and weight loss. At necropsy, the bird was emaciated and had reddened and wet lungs. Microscopic lesions were limited to hepatic and pulmonary congestion with focal thrombosis. Erysipelothrix rhusiopathiae was isolated by routine bacterial culture from several organs. Further characterization of the isolate by pulsed-field gel electrophoresis indicated that the isolate has a new genotype pattern 3A(III), which is 91.7% homologous to an E. rhusiopathiae that was isolated from a pig in 2001 and 88% homologous to an isolate recovered in 2000 from a turkey with septicemia. This is the first report of E. rhusiopathiae induced septicemia in a kookaburra. PMID- 16312249 TI - Isolation of equine herpesvirus-2 from the lung of an aborted fetus. AB - This study describes the isolation of equine herpesvirus-2 (EHV-2) from the lung of an aborted equine fetus in Argentina. The isolated virus was confirmed as EHV 2 by indirect immunofluorescence using a rabbit anti-EHV-2 polyclonal antiserum and by virus-neutralization test using an equine polyclonal antibody against EHV 2. Restriction endonuclease DNA fingerprinting with BamHI also confirmed the identity of the virus as EHV-2. Furthermore, viral nucleic acid was detected by polymerase chain reaction from the original lung sample and from the DNA obtained from cells infected with the virus isolate. This work constitutes the first reported isolation of EHV-2 from an aborted equine fetus. The presence of EHV-2 in the lung of the aborted fetus would indicate that this virus is capable of crossing the placental barrier. However, no cause-effect relationship was established between the EHV-2 isolate and the abortion. PMID- 16312250 TI - Begin with giving. PMID- 16312251 TI - Water availability and usage on the New Mexico/Mexico border. AB - New Mexico, one of four states on the U.S./Mexico border, is faced with a pressing concern--lack of water. Since the region is either arid or semiarid, it is chronically short of continually available surface-water resources. Groundwater resources are used beyond their capacity to be recharged, and most surface-water resources are used to the maximum. The quality of groundwater varies widely. As a result of nonpoint- and point-source contamination, as well as natural occurrence, water in some areas is too salty or has high levels of natural uranium, fluoride, or arsenic. To date, the New Mexico Environment Department (NMED) has recognized 1,400 cases of groundwater contamination, and 1,907 water supply wells have been affected (NMED, 2001a). Of approximate 4,000 miles of coninously flowing rivers and streams in New Mexico, 92 perent are affected by nonpoint sources of pollution (NMED, 2001b). Numerous critical water issues exist along the New Mexico/Mexico border as a result of the impending critical issue of water availability, usage, and quality, as well as the fast growing population. Related public health problems along the New Mexico/Mexico border are indicative of the need for a holistic, concrete, and sustainable solution to meet water demands in New Mexico. In order to accomplish the goals an objectives of Border XXI, Healthy People 2010, and Heathy Border 2010, a comprehensive statewide water management plan is needed. Solutions to the water demands of the region will be addressed in a subsequent manuscript. PMID- 16312252 TI - Assessing the effectiveness of food worker training in Florida: opportunities and challenges. AB - The task of measuring the effectiveness of food worker training has historically met with many challenges. This paper considers various approaches and utilizes trends in foodborne-outbreak contributing factors to evaluate a recent change in Florida's food worker training. Results show that subsequent to training, the relative incidence of many factors that contribute to foodborne outbreaks actuay increased, while the relative incidence of other factors decreased. The overall rate of foodborne outbreaks associated with the contributing factors that the authors studied decreased subsequent to training. Results of this analysis must be interpreted with caution because of multiple confounding factors; however, it became apparent that both increases and decreases in the occurrence of contributing factors could be used to focus future training material on areas of food handler practices in which it is needed. Further work needs to be done to estabish the most useful methods and approaches for assessing effectiveness and hence the public health impact of food worker training. PMID- 16312253 TI - Food safety issues and training methods for ready-to-eat foods in the grocery industry. AB - As Americans have become more pressed for time, the use of convenient, simplified meals become a way of life. One aspect of this trend, known as Home Meal Replacement (IIMR), has increased in sales since its inception. Between 1999 and 2001, the average annual expenditure per consumer rose 5.6 pereent, and $958 per person per year was spent in 2002. Along with this growth, food safety risks may have increased. The study reported here examined efforts being undertaken by grocery and convenience stores to control the wholesomeness of INR food items. After a convenience sample of 500 grocery store executives was identified, a 32 item questionnaire was developed and mailed to the executives. The results indicate that the industry has taken food safety seriously with only 10 pereent reporting that they have no food safety training. The executives cited employee turnover as a major concern in food safety today, along with lack of food safety knowledge of the consumer and improper holding temperatures. PMID- 16312254 TI - A new alternative for sizing septic systems. PMID- 16312255 TI - Avian influenza and the significance of its transmission to humans--information from WHO. PMID- 16312257 TI - NEHA's 2005 annual educational conference & exibition: advancing the environmental health profession. PMID- 16312256 TI - Impact of restaurant hygiene grade cards on foodborne-disease hospitalizations in Los Angeles County. PMID- 16312258 TI - My AEC awards! PMID- 16312259 TI - Quantitative immunoelectron-microscopic analysis of the type IV collagen alpha1-6 chains in the glomerular basement membrane in childhood thin basement membrane disease. AB - AIM: Thin basement membrane disease (TBMD) is characterized histologically by diffuse thinning of glomerular basement membrane (GBM). Although recent genetic analysis has shown that TBMD might be included within type IV collagen disorders, conventional immunohistochemical studies demonstrated normal labeling of type IV collagen alpha chains in the GBM. We have, however, successfully used confocal laser scanning microscopy to demonstrate a significantly reduced signal of type IV collagen alpha5 chain (alpha5(IV)) along capillary walls in TBMD. In order to further understand the association of type IV collagen with TBMD, we used immunoelectron microscopy to examine renal biopsies from 6 children with TBMD and six control children with minimal change nephrotic syndrome. METHODS: Ultrathin sections of LR gold resin were incubated with a rat monoclonal antibody against human alpha1(IV), alpha2(IV), alpha3(IV), alpha4(IV) alpha5(IV) or alpha6(IV) followed by colloidal gold conjugated goat anti-rat IgG. After taking electron micrographs, the labeling was quantitatively evaluated in the area occupied by the segments of basement membrane. The basement membrane was divided into three equal segments viz. subepithelial side, central portion and subendothelial side. RESULTS: In control subjects, the number of gold particles for alpha1(IV) or alpha2(IV) was significantly greater in the subendothelial side and central portion than in the subepithelial side of the GBM, whilst alpha3(IV), alpha4(IV) or alpha5(IV) labeling was significantly more prominent in the central portion compared to the subepithelial and subendothelial side of the GBM. TBMD samples showed a similar distribution pattern except that the subepithelial side and central portion of the GBM had a significantly reduced amount of alpha5(IV) antigen compared to control subjects. CONCLUSION: This is the first report demonstrating a diminished labeling intensity of alpha5(IV) in the central portion and subepithelial side of the GBM in renal biopsy specimens from patients with TBMD. These findings suggest that an abnormality of alpha5(IV) might possibly be associated with the pathogenesis of TBMD. PMID- 16312260 TI - Chromosomal telomere shortening of kidney cells in IgA nephropathy by the measurement of DNA in urinary sediment. AB - BACKGROUND: The histology and function of the kidney deteriorates with age and progressive renal failure, but the mechanisms involved in renal ageing are not known. In vitro studies suggest that telomere shortening is important in replicative senescence, and is accelerated by stress factors that increase replication. We investigated whether IgA nephropathy, a prototype chronic kidney disease, is associated with localized intrarenal cellular ageing. METHODS: We studied the mean length of terminal restriction fragments (TRF), a measure of average telomere size, in the DNA of peripheral blood mononuclear cells and urinary sediment of 15 patients with IgA nephropathy. RESULTS: The mean TRF lengths in peripheral blood is 7043.8 +/- 1 182.8 base pairs, and in urinary sediment is 6 749.7 +/- 636.5 base pairs. The mean TRF lengths of urinary DNA significantly correlate with the serum creatinine (r = -0.525, p = 0.044) and estimated glomerular filtration rate (GFR) (r = 0.651, p = 0.009). The mean TRF lengths of urinary DNA had an insignificant inverse correlation with patient age (r = -0.364, p = 0.2), and do not correlate with the degree of glomerulosclerosis (r = 0.004, p = 0.9) or tubulointerstitial scarring in renal biopsy (r =-0.032, p = 0.9). After 30 months of follow-up, the rate of decline of estimated GFR has an inverse correlation with the mean TRF lengths of urinary DNA (r = -0.699, p = 0.004). The TRF lengths of peripheral blood DNA do not correlate with any clinical or histological parameter or the rate of renal function decline. CONCLUSIONS: Although this is a pilot study, our observation indicates that the TRF lengths of genomic DNA extracted from urinary sediment is related to the degree of renal impairment. However, a long telomere length of genomic DNA in urinary sediment is associated with a more rapid decline of renal function. Our findings might be relevant to the pathogenesis of progressive renal failure. PMID- 16312261 TI - Oxidative stress and non-enzymatic glycation in IgA nephropathy. AB - AIM: Approximately 20-50% of IgA nephropathy patients develop end-stage renal disease. We have previously found enhanced oxidative stress and decreased antioxidant capacity in red blood cells of IgA nephropathy patients. In this study we assess oxidative stress, non-enzymatic glycation, oxidative resistance of low-density lipoprotein and its alpha-tocopherol content in these patients. PATIENTS AND METHODS: Non-enzymatic glycation and oxidative stress were assessed in 88 IgA nephropathy patients by measuring advanced glycation end products, Nepsilon-carboxymethyl-lysine, thiobarbituric acid reactive substances, oxidative resistance of low-density lipoprotein and its alpha-tocopherol content. RESULTS: Advanced glycation end products (2659 +/- 958 a.u.) and Nepsilon-carboxymethyl lysine (563 +/- 215 ng/ml) were significantly higher in IgA nephropathy patients with decreased renal function compared to those with normal renal function (p < 0.002) or controls (p < 0.001). Thiobarbituric acid-reactive substances in plasma and associated with low-density lipoprotein were significantly elevated and oxidative resistance of low-density lipoprotein was significantly reduced in all groups of IgA nephropathy patients. There was no significant difference in circulating fluorescent advanced glycation end products, Nepsilon-carboxymethyl lysine, thiobarbituric acid-reactive substances levels, oxidative resistance of low-density lipoprotein and its alpha-tocopherol content between patients with normal vs. impaired glucose metabolism. Low alpha-tocopherol content of low density lipoprotein was accompanied with decreased oxidative resistance, depletion in polyunsaturated fatty acids, elevated saturated fatty acids and thiobarbituric acid-reactive substances within low-density lipoprotein suggesting enhanced lipid peroxidation. CONCLUSIONS: Decreased oxidative resistance of low density lipoprotein and enhanced oxidative stress are common features in IgA nephropathy, while increased non-enzymatic glycation occurs as renal function declines. PMID- 16312262 TI - Evaluation of intrarenal hemodynamics by Doppler ultrasonography for renoprotective effect of angiotensin receptor blockade. AB - AIMS: It has been shown that both angiotensin-converting enzyme inhibitors (ACE I) and angiotensin II type 1 receptor blockers (ARB) have renoprotective effects via mechanisms that are independent of blood pressure reduction. The aim of this study was to evaluate the intrarenal hemodynamic change with ARB by renal Doppler ultrasonography (RDU) and to assess the mechanism of ARB in patients with hypertension. METHODS: Thirty hypertensive patients with renal insufficiency caused by glomerular diseases, diabetes and hypertensive nephrosclerosis were included in this study. RDU was performed before and one week after taking ARB. Resistance index (RI) (peak systolic velocity - end diastolic velocity/peak systolic velocity) in the intrarenal segmental artery were calculated, and the amounts of urinary protein or albumin were determined. RESULTS: We defined patients whose microalbuminuria or proteinuria was reduced by greater than 30% by ARB as responders (n = 22) and defined other patients as non-responders (n = 8). There were no significant differences between the responder and non-responder groups in baseline characteristics. RI was significantly improved by ARB in the responder group, but not in the non-responder group. The reduction of RI after ARB treatment was most prominent in patients with hypertensive nephrosclerosis. CONCLUSIONS: Improvement in intrarenal hemodynamics might play an important role in the mechanisms of the renoprotective effect of ARB in patients with hypertension. PMID- 16312263 TI - The effect of angiotensin receptor blockade ARB on the regression of left ventricular hypertrophy in hemodialysis patients: comparison between patients with D allele and non-D allele ACE gene polymorphism. AB - OBJECTIVE: It is revealed that LVH is one of risk factors for the development of cardiac complications in long-term HD patients. Therefore, maneuvers to reduce hypertrophy of cardium are very important for improving life prognosis. Angiotensin II receptor blockade (ARB) could reduce LVH in general populations without renal failure. However, no conclusive data has been available regarding the clinical consequences of ARB administration on the regression of LVH in HD patients. Furthermore, it has not clearly determined if ACE gene polymorphism has a possible influential effect on it. This study is conducted to clarify these issues. SUBJECTS AND METHOD: 32 hypertensive patients on regular HD (male/female: 21/11, mean age: 60.5 years, mean duration of HD: 52.8 months) were studied. Patients were classified into two groups according to the different type of ACE gene polymorphism: cases with D allele (DD/ID; D group: n = 13) and those without (II; non-D group: n = 19). All patients were administered ARB (losartan 50 - 100 mg/day) and echocardiography (UCG) was performed at 6-month-interval regularly until the end of observation (24 months). RESULTS: Before the commencement of ARB, no differences were found between the two groups, neither in mean blood pressure (MBP: D group/non-D group: 120 +/- 13 vs. 115 +/- 14 mmHg) nor in left ventricular mass index (LVMI: D/non-D: 172 +/- 41 vs. 165 +/- 41 g/m2). During the 24r-month follow-up, there were significant and similar reductions in MBP in both groups. In respect to LVMI, a significant reduction of LVMI was found in the D group after six months (p < 0.01 vs. basal) with a final reduction rate (FRR) 26 +/- 13%, whereas in the non-D group it was found at 24 months (p < 0.01 vs. basal) with FRR -11 +/- 16% (p < 0.01 vs. D group). There were significant differences between the two groups at all points (p < 0.05 at 6, 18 and 24 months, p < 0.005 at 12 months, respectively). CONCLUSION: It is indicated that ARB could insert a regression effect on LVH predominantly in patients with D allele ACE polymorphism, due partly to factor (s) independent of its anti hypertensive effect. PMID- 16312264 TI - Improvements in phosphate control with short daily in-center hemodialysis. AB - BACKGROUND: Hyperphosphatemia is an independent risk factor for mortality in hemodialysis (HD) patients. The relative importance of HD frequency and duration for phosphate removal is not clear. Short daily hemodialysis (SDHD) is a form of HD which offers increased treatment frequency. SDHD studies have not been shown to normalize serum phosphate. METHODS: Twenty-one patients were converted from conventional thrice weekly HD (CHD, 4 h/session) to SDHD (2 - 3.75 h/session, 5 - 6 sessions per week). The primary endpoint was the change in predialysis serum phosphate levels after conversion from CHD to SDHD. Changes in serum calcium levels, phosphate binder and vitamin D analogue usage, and serum parathyroid hormone (PTH) levels were measured as secondary endpoints. RESULTS: Mean duration of SDHD was 17.7 +/- 1.9 months. Mean treatment time was 2.63 +/- 0.10 h, and mean frequency was 5.3 +/- 0.1 sessions per week. Predialysis serum phosphate decreased from 1.99 +/- 0.12 mM at three months pre conversion to 1.27 +/- 0.10 mM at six months post conversion to SDHD (p = 0.002). Serum phosphate remained stable between six and 12 months post conversion (1.27 +/- 0.10 mM to 1.38 +/- 0.14 mM, p = 0.8). When patients were grouped according to SDHD sessional frequency (five sessions/week versus six sessions/week) and compared, no significant differences were found in predialysis serum phosphate levels at six or 12 months post conversion. There were no changes in serum calcium. Overall phosphate binder usage did not change pre and post conversion to SDHD. Serum PTH tended to decrease after one year of SDHD (44.2 +/- 13.4 pM to 21.4 +/- 5.9 pM, p = 0.07). CONCLUSION: Conversion to SDHD significantly decreased serum phosphate. There may be a minimum hemodialysis duration below which increases in frequency are not able to compensate to achieve normal phosphate levels. Future studies are necessary to better characterize the relationship between HD duration and frequency with respect to phosphate removal. PMID- 16312265 TI - Underutilization of aspirin in hemodialysis patients for primary and secondary prevention of cardiovascular disease. AB - BACKGROUND: Patients on hemodialysis are at high risk for cardiovascular disease (CVD). Aspirin is an established therapy for primary and secondary prevention of CVD that may be underutilized in hemodialysis patients. To better understand the use of aspirin in hemodialysis patients, we examined the experience of an urban hemodialysis center. Guidelines for use as well as associated risks and benefits are reviewed. METHODS: Medical records for patients receiving hemodialysis treatment at our center (New York City, USA) in May 2004 were reviewed for aspirin use, presence of CVD, and potential contraindications to aspirin therapy. CVD was defined as a history of coronary artery disease, ischemic stroke, transient ischemic attack, or peripheral vascular disease. Potential contraindications to aspirin therapy included history of clinically significant bleeding or increased risk of bleeding, aspirin allergy and routine treatment with other anticoagulants. RESULTS: 176 patients were eligible for the study and 172 (98%) were included. Although 74 patients had a history of CVD, only 38 (51 %) of these were treated with aspirin. Among patients with a history of CVD who were not treated with aspirin, 19 (53%) had no identifiable contraindications to aspirin therapy for secondary prevention of CVD. Ninetyeight patients had no history of CVD, and 18 (18%) of these were treated with aspirin. Of patients without a history of CVD who were not treated with aspirin, 57 (71%) had no identifiable contraindications to aspirin therapy for primary prevention of CVD. CONCLUSIONS: Aspirin is underutilized in hemodialysis patients for the primary and secondary prevention of CVD. Given the high risk of CVD in hemodialysis patients, therapy with aspirin may be of significant benefit and prospective studies of aspirin therapy are needed. PMID- 16312266 TI - Predictive factors of outcome following staphylococcal peritonitis in continuous ambulatory peritoneal dialysis. AB - BACKGROUND AND AIMS: Staphylococcus epidermidis and other coagulase-negative staphylococci (CoNS) are the most common agents of continuous ambulatory peritoneal dialysis (CAPD) peritonitis. Episodes caused by Staphylococcus aureus evolve with a high method failure rate while CoNS peritonitis is generally benign. The purpose of this study was to compare episodes of peritonitis caused by CoNS species and S. aureus to evaluate the microbiological and host factors that affect outcome. MATERIAL AND METHODS: Microbiological and clinical data were retrospectively studied from 86 new episodes of peritonitis caused by staphylococci species between January 1996 and December 2000 in a university dialysis center. The influence of microbiological and host factors (age, sex, diabetes, use of vancomycin, exchange system and treatment time on CAPD) was analyzed by logistic regression model. The clinical outcome was classified into two results (resolution and non-resolution). RESULTS: The odds of peritonitis resolution were not influenced by host factors. Oxacillin susceptibility was present in 30 of 35 S. aureus lineages and 22 of 51 CoNS (p = 0.001). There were 32 of 52 (61.5%) episodes caused by oxacillin-susceptible and 20 of 34 (58.8%) by oxacillin-resistant lineages resolved (p = 0.9713). Of the 35 cases caused by S. aureus, 17 (48.6%) resolved and among 51 CoNS episodes 40 (78.4%) resolved. Resolution odds were 7.1 times higher for S. epidermidis than S. aureus (p = 0.0278), while other CoNS had 7.6 times higher odds resolution than S. epidermidis cases (p = 0.052). Episodes caused by S. haemolyticus had similar resolution odds to S. epidermidis (p = 0.859). CONCLUSIONS: S. aureus etiology is an independent factor associated with peritonitis non-resolution in CAPD, while S. epidermidis and S. haemolyticus have a lower resolution rate than other CoNS. Possibly the aggressive nature of these agents, particularly S. aureus, can be explained by their recognized pathogenic factors, more than antibiotic resistance. PMID- 16312267 TI - Interstitial nephritis and high titers of PR3-ANCA: an unusual manifestation of ANCA-associated disease. AB - We present the case of a 75-year-old female with weight loss, anemia, systemic signs of inflammation, mild renal insufficiency, microscopic hematuria, mixed glomerular and tubular proteinuria, and high titers of PR3-ANCA. Renal biopsy demonstrated interstitial nephritis with some sclerosed but otherwise normal glomeruli. Extensive work-up showed no signs of granulomatous inflammation or other vasculitic organ involvement. We presumed this to be a rare renal manifestation of ANCA-associated disease with the presence of sclerosed glomeruli suggesting a previous history of glomerular involvement. In view of the absence of active vasculitic or granulomatous disease, treatment was limited to low-dose corticosteroids with good response. PMID- 16312268 TI - Tubulointerstitial macrophage infiltration in a patient with hypokalemic nephropathy and primary Sjogren's syndrome. AB - We report a case of hypokalemic nephropathy associated with primary Sjogren's syndrome (SS). The patient presented with profound and persistent hypokalemia secondary to distal renal tubular acidosis (RTA). A renal biopsy exhibited tubular degeneration, marked interstitial fibrosis and intense macrophage infiltration. Hypokalemia has been reported to induce macrophage infiltration in experimental animal models but not in humans. This is the first report of intense tubulointerstitial macrophage infiltration in a patient with hypokalemic nephropathy associated with SS. PMID- 16312269 TI - A case of rhabdomyolysis induced acute renal failure secondary to statin-fibrate derivative combination and occult hypothyroidism. AB - Rhabdomyolysis is a syndrome characterized by muscle necrosis and the release of intracellular muscle contents into the systemic circulation. We hereby report a patient proved to be a case of unrecognized hypothyroidism presenting with rhabdomyolytic acute renal failure precipitated by the combined use of statin and fenofibrate. A 63-year-old woman was referred to our department because of fatigue, diffuse muscle pain and oliguria. On the basis of pathogenesis, clinical and laboratory examination the diagnoses of acute renal failure secondary to the statin-fibrate-derivative combination induced rhabdomyolysis and auto-immune thyroiditis induced hypothyroidism were made. Although saline, furosemide and sodium bicarbonate infusions enabled diuresis and have led to a rapid recovery of renal function and normalization of blood pressure in five days (creatinine level decreased from 4.5 mg/dl to 1.2 mg/dl), only thyroid replacement therapy (0,1 mg thyroxine) that begun after the exclusion of adrenal insufficiency resulted in complete resolution of rhabdomyolysis. This prompted the diagnosis of background, clinically silent rhabdomyolysis aggrevated by the statin-fibrate-derivative combination. To our knowledge this case illustrates the first example of rhabdomyolytic acute renal failure induced by a statin-fibrate-derivative combination with underlying hypothyroidism which was responsible for the basal clinically unobservable rhabdomyolysis. PMID- 16312270 TI - High serum estradiol concentrations in postmenopausal women with end-stage renal disease. PMID- 16312271 TI - Rapidly progressive glomerulonephritis in a patient with Waldenstrom's macroglobulinemia. PMID- 16312272 TI - [Incidence of endometriosis in infertile women: clinical and laparoscopic characteristics]. AB - BACKGROUND: The incidence and symptomatology of endometriosis vary according to the different populations. OBJECTIVES: To determine the incidence of endometriosis as well as the clinical and laparoscopic characteristics in infertile women with endometriosis. PATIENTS AND METHODS: This study included 68 infertile patients with endometriosis diagnosed by laparoscopy; in all of them we analyzed the demographic and clinical characteristics. RESULTS: In 68 out of 197 medical files that were analyzed, endometriosis was reported (34.5%). The average age was 30.3 +/- 3.9 years. Forty patients had primary infertility (58.8%) and 28 (41.29%) secondary infertility. In 34 patients endometriosis (50%) was mild, moderate in 16 (23.5%) and severe in 18 (26.5%). Only one endometriosic foci was found in 25% of the patients and multiple foci in 75% of them. The most common sites in which it was found were the uterosacral ligaments, Douglas pouch and the ovaries. In patients with primary infertility it predominated the medium-high socioeconomic level and in those with secondary infertility the low one. In severe endometriosis the predominant symptom was the moderate or severe dysmenorrhea, while in mild and moderate endometriosis was mild dysmenorrhea (p < 0.007). Six out of 16 patients with severe endometriosis had endometrioma. CONCLUSIONS: The endometriosis incidence in infertile women was similar to that reported in literature, as well as age of presentation. It was also observed a direct relationship between endometriosis severity and dysmenorrhea intensity. PMID- 16312273 TI - [Analysis of trends in maternal mortality during a 10 year-follow up in a urban region]. AB - BACKGROUND: Maternal mortality is considered a worldwide health problem, since the WHO has estimated that there are 500,000 maternal deaths annually and that the highest percentage of these deaths occurs in developed countries. It was not until 1970 that this problem became important, by revising the birth's registrations and the maternal causes of morbidity and mortality, and that the mortality committees study began. OBJECTIVE: To determine the epidemiological profile and the trend of the maternal mortality in a urban hospital. MATERIAL AND METHODS: This retrospective and descriptive study was carried out at the Reproductive Health Coordination of the Centro Medico Nacional La Raza, through the registrations of the maternal deaths reported from 1993 to 2002 by the local committees of maternal mortality. Maternal mortality was defined as an event occurred during pregnancy or within 42 days after delivery, by any associated cause and aggravated by the same pregnancy. We used analytic and descriptive statistics, with central trend measurements, as well as chi square test. RESULTS: During the 10 years of analysis, 244 deaths were shown, with mean per year of 24.4 +/- 5.2 deaths. Preeclampsia-eclampsia, hypovolemia and sepsis represented 71.7% of the cases. The highest number of deaths ranged from 30 to 34 years, with 64 deaths. Women with the first pregnancy had the highest number of deaths, with incidence of one maternal death for each 664.7 +/- 135.6 new born in each year. DISCUSSION: The main maternal deaths per year corresponded to direct obstetric causes, with percentages from 53.3 to 86.9% annually, where preeclampsia eclampsia syndrome maintains these high percentages, with a death average within the international standard parameters considered adequate. PMID- 16312274 TI - [Gestational diabetes mellitus. Experience at a third level hospital]. AB - BACKGROUND: The prevalence of type 2 diabetes mellitus and of gestational diabetes mellitus is high in the Mexican population; thus, strategies to improve its detection and prevent obstetric and perinatal complications are essential. PATIENTS AND METHODS: During the period 2000-2004 a total of 8,074 pregnant women were studied from the day of performance, 50-g, 1 hour glucose screening test for gestational diabetes mellitus (ST-GDM) until the end of pregnancy using the same protocol: ST-GDM was performed immediately after 14 weeks of gestation in high risk women and between 24-28 weeks in those with regular risk. Two weeks later 100-g, 3 hour oral glucose tolerance test (3h-OGTT) was performed in women with ST-GDM [symbol: see text] 130 but < 180 mg/dL, and patients were classified according to Freinkel's criteria in class A1, A2 and B1 gestational diabetes mellitus. All women received a diet according to their ideal weight for gestational age and, when necessary, insulin was added to achieve an adequate glycemic control. RESULTS: From 8,074 glucose screening tests for gestational diabetes mellitus, 37.2% (n = 2,997) were positive and 17.2% (n = 514) diagnostic for gestational diabetes mellitus (serum glucose [symbol: see text] 180 mg/dL). In 2,483 patients, ST-GDM was positive but negative for gestational diabetes (serum glucose >130 but <180 mg/dL); in 1,070 of them (43.0%) gestational diabetes mellitus was diagnosed by means of 3h-OGTT. The diagnosis of class A1 gestational diabetes mellitus was done in 908 patients (84.9%), A2 in 188 (11.8%) and B1 in 51 (3.2%). Patients with gestational diabetes diagnosed only by an abnormal ST-GDM were classified as class A1 gestational diabetes mellitus. Gestational age at diagnosis by ST-GDM was 12.4 +/- 4.7 weeks and 27.4 +/- 5.8 weeks when diagnosed by a 3h-OGTT. The total daily dose of insulin required was related to Freinkel's classification: 16.0% (n = 215/1,345) with class A1 gestational diabetes mellitus required 0.48 +/- 0.33 U/day/kg and 85.0% (n = 43/51) with class B1 needed 1.0 +/- 0.32 U/day/kg at the end of pregnancy, which occurred at 38.2 +/- 2.0 weeks in all 3 groups. In decreasing order of occurrence perinatal complications were: pregnancy-induced hypertension (specially in class B1 gestational diabetes mellitus), urinary tract infections and premature rupture of membranes. Birth weights either < 2,500 g or > 4,000 g were more frequent among women with class B1 gestational diabetes, than in the other 2 groups. After pregnancy ended (6-8 weeks), 52.2% of women with gestational diabetes (mostly class B1) was diagnosed with permanent type 2 diabetes mellitus. DISCUSSION: In this study the percentage of pregnant women diagnosed as having gestational diabetes mellitus exclusively by ST-GDM was high (17.2%). Freinkel's classification of such disease predicted higher risk in women with class B1 gestational diabetes mellitus for: 1) obstetric complications, 2) higher doses of insulin to achieve an adequate glycemic control; 3) newborns' birth weight < 2,500 g or > 4,000 g for gestational age, 4) having type 2 diabetes mellitus in late puerperium. PMID- 16312275 TI - [Endometriosis. Is it a problem of the immunological signs?]. AB - Endometriosis syndrome affects 5% to 35% of women in reproductive age. Many theories have been suggested, but the true importance is the survival mechanism of endometriotic tissue, which seems to be related to a defect of the immunological vigilance from macrophages, which are responsible for secretion of a variety of biochemical substances such as cytokines, prostaglandins and growth factors, which through different ways stimulates the growth of the endometrial cells. This has been observed for the clear interrelation between the homeostasis of the body and the evident association of the hormonal factors with the processes of the immunological system, in whose field many steps are unknown. PMID- 16312276 TI - [Spermatogenesis arrest]. AB - Spermatogenesis arrest is a complex process of interruption in the differentiation of germinal cells of specific cellular type, which elicits an altered spermatozoa formation. In contrast, hypospermatogenesis is defined as a decrease in number of germ cells and its proportion. Factors identified intervening upon spermatogenesis arrest are: genetic, hormonal, growth factors, interaction between Sertoli and germ cells and ectoplasmic specialization integrity of spermatozoa. In addition, environmental toxic effects have shown to exert subletal and letal cellular damage with gene disruption. Hence in this work we review sperm physiology along with etiologic elements associated to spermatogenesis arrest delineating the most appropriate conduct for diagnosis and treatment. PMID- 16312277 TI - [Effects of splenic function and splenic diseases on female physiology, with a brief analysis of 182 gestations after splenectomy. 1950]. PMID- 16312278 TI - Evaluation of bacterial sensitivity of chemoantibiotics by means of two semiautomatic systems. Cobas-bact and TTS. PMID- 16312279 TI - A simple, totally implantable infusion system for long-term drug treatment in the laboratory rat. PMID- 16312280 TI - Influence of antibiotics on adhesion of bacteria. PMID- 16312281 TI - The influence of foreign bodies on the in vitro activity of various antibiotics against S. aureus, S. epidermidis and gram-negative strains. PMID- 16312282 TI - Effect of antimicrobial agents on the oral bacteria coaggregation. PMID- 16312283 TI - Evaluation of rapid ATB: a system for 4h antimicrobial susceptibility testing. PMID- 16312284 TI - Variations of autochthonous skin flora in hospitalized patients. PMID- 16312285 TI - Staphylococci in Spain. PMID- 16312286 TI - In vitro antibiotic susceptibilities of Streptococcus viridans blood isolates from neutropenic patients. PMID- 16312287 TI - Group A streptococcal tonsillitis: comparative evaluation of kits for rapid diagnosis. PMID- 16312288 TI - A murine model of systemic infection with serotype IV group B streptococcus. PMID- 16312289 TI - Blood isolates in a general hospital: species and susceptibility analysis. PMID- 16312290 TI - Virulence factors of Haemophilus influenzae. PMID- 16312291 TI - Improved susceptibilities to cinoxacin and other antimicrobial agents of some Providencia species re-identified on the basis of fermentation of polyhydric alcohols. PMID- 16312292 TI - The microbial spectrum and the importance of Gram's stain in external otitis and in otitis media. PMID- 16312293 TI - Isolation of Campylobacter pylori in patients with non-ulcer dyspepsia. PMID- 16312294 TI - Italian Haemophilus influenzae surveillance results. PMID- 16312295 TI - Non-influence of topical chloramphenicol treatment on the diagnosis of chlamydial conjunctivitis by culture and direct specimen antigen detection tests. PMID- 16312296 TI - Endotoxin and coagulation disorders: an experimental approach. PMID- 16312297 TI - A new mouse model of Chlamydia trachomatis MoPn genital infection. PMID- 16312298 TI - Identification of a trypanocidal factor in normal human sera. PMID- 16312299 TI - Assessments of drug interactions in vitro. PMID- 16312300 TI - Characterization of commercial lots of ampicillin sodium raw material. PMID- 16312301 TI - In vitro antibacterial activity of lenampicllin. PMID- 16312302 TI - Bactericidal activity and morphological effects of lenampicillin on Staphylococcus aureus. PMID- 16312303 TI - Comparative in vitro activity of imipenem and nine other antimicrobials against recently isolated pathogens. PMID- 16312304 TI - Pharmacodynamic determinants of imipenem's efficacy in a man-adapted animal model. PMID- 16312305 TI - Antimicrobial activity of imipenem against human anaerobic bacteria isolated in Rio de Janeiro, Brazil. PMID- 16312306 TI - The postantibiotic effect (PAE) of the new penem FCE 22101 on gram-positive and negative bacteria in vitro and in vivo. PMID- 16312307 TI - Comparative activities in vitro of amoxycillin/clavulanic acid and ampicillin/sulbactam. PMID- 16312308 TI - Comparative efficacy of amoxycillin + clavulanic acid (augmentin), flucloxacillin and vancomycin against S. aureus endocarditis in the rat. PMID- 16312309 TI - Comparative activities in vitro of augmentin (amoxycillin/clavulanic acid) and oral cephalosporins. PMID- 16312310 TI - Bacteriological studies of amoxycillin-clavulanic acid against anaerobic bacteria. PMID- 16312311 TI - Kinetics of antimicrobial activity of amoxycillin/clavulanic acid against Bacteroides fragilis group. PMID- 16312312 TI - Effect of amoxycillin/clavulanic acid (augmentin) against polymicrobial Bacteroides fragilis/Escherichia coli infections in the mouse. PMID- 16312313 TI - Distribution and efficacy studies with ticarcillin-clavulanic acid (timentin) in experimental Klebsiella pneumoniae meningitis in rabbits. PMID- 16312314 TI - Synergy between ticarcillin/clavulanic acid and aminoglycosides against Pseudomonas aeruginosa infections in neutropenic mice. PMID- 16312315 TI - Bactericidal effects of amoxycillin/clavulanic acid (augmentin) against Legionella pneumophila in vitro and in tissue culture studies. PMID- 16312316 TI - Piperacillin + tazobactam: fixed ratio versus fixed concentration synergism studies. PMID- 16312317 TI - YTR-830H: its origin, the present and the future. PMID- 16312318 TI - In vitro evaluation of new beta-lactamase inhibitor: YTR830H and derivatives. PMID- 16312319 TI - In vitro activity of YTR-830H in combination with beta-lactamase unstable beta lactams. PMID- 16312320 TI - Comparison of the antimicrobial activity of cefuroxime with other oral antibiotics. PMID- 16312321 TI - Worldwide assessment of the activity of cefotetan against clinical isolates. PMID- 16312322 TI - Cefotetan: antibacterial activity against Staphylococcus aureus in the presence of human serum. PMID- 16312323 TI - Serum bactericidal activity against aerobes and anaerobes of volunteers receiving cefoxitin or cefotetan. PMID- 16312324 TI - In vitro study of the antibacterial activity of cefotetan compared with piperacillin and ceftriaxone against bacterial strains isolated from pathological material of hospital patients. PMID- 16312325 TI - Synergistic activity of piperacillin: YTR-830H combinations in vitro and in vivo. PMID- 16312326 TI - The in-vitro activity of three long-acting cephalosporins against Bacteroides fragilis, Peptostreptococcus species and Clostridium perfringens. PMID- 16312327 TI - New trends in oral cephalosporin research. PMID- 16312328 TI - Worldwide review: microbiological profile of cefixime. PMID- 16312329 TI - Cefixime versus cefotaxime against gram-negative bacteria causing urinary tract infections. PMID- 16312330 TI - An epidemiological study of three cephalosporins: cefotetan, cefixime and cefpirome. PMID- 16312331 TI - The versatile monobactam family: manipulation of intrinsic activity and in vivo efficacy via N-1 activating group modification. PMID- 16312332 TI - Comparative in vitro antibacterial activities of cefetamet (Ro 15-8074/005), a new oral cephalosporin, and eight other oral antimicrobials. PMID- 16312333 TI - In vitro evaluation of Ly 163892, a new oral cephalosporin. PMID- 16312334 TI - In vitro and in vivo experience with carumonam: a preliminary report. PMID- 16312335 TI - Comparative antibacterial activity of the new oral monobactam tigemonam (SQ 30836) against urinary isolates. PMID- 16312336 TI - Combination of monobactams and other antibiotics. PMID- 16312337 TI - Polarography of 4-quinolones, their determination and metal complex formation study. PMID- 16312338 TI - The versatile monobactam family: new agents addressing specific needs in infectious disease. PMID- 16312339 TI - Interactions between 4-quinolone antibacterials and multivalent metal ions. PMID- 16312340 TI - In vitro activity of the new quinolones: an overview. PMID- 16312341 TI - Effect of fluorinated quinolones on heat-labile toxin production by Escherichia coli. PMID- 16312342 TI - Enhancement of antibacterial activity of norfloxacin and ofloxacin in vitro by serum and experimental pleural exudates. PMID- 16312343 TI - Fluoroquinolones: possible additive effects in combinations against problem bacteria. PMID- 16312344 TI - Antimicrobial activity of enoxacin. PMID- 16312345 TI - Evaluation of ofloxacin activity against recent clinical isolates. PMID- 16312346 TI - Effects of sublethal concentrations of pefloxacin and ofloxacin on the adherence of Escherichia coli strains to human uroepithelial cells in vitro. PMID- 16312347 TI - Determination of the postantibiotic effect of ofloxacin on gram-negative bacteria utilizing a new method. PMID- 16312348 TI - The bactericidal activity of ofloxacin against staphylococci. PMID- 16312349 TI - In vitro comparison of ciprofloxacin to parenteral antibiotics. PMID- 16312350 TI - Ciprofloxacin in vitro activity alone and in combination against clinical isolates of glucose nonfermenting bacteria. PMID- 16312351 TI - Antimicrobial activity of a new difluorinated quinolone: lomefloxacin (SC-47111). PMID- 16312352 TI - In vitro activity and synergy studies with fleroxacin. PMID- 16312353 TI - In vitro evaluation of lomefloxacin (NY-198 or SC-47111), a new difluoro quinolone. PMID- 16312354 TI - Microbiological studies on lomefloxacin--a new quinolone. PMID- 16312355 TI - Preliminary results on the antibacterial and killing activity of lomefloxacin. PMID- 16312356 TI - In vitro antimicrobial activity of lomefloxacin against staphylococci, enterococci, and Enterobacteriaceae. PMID- 16312357 TI - In vitro activity of lomefloxacin, a new difluorinated quinolone, against urinary bacterial isolates: comparison with enoxacin and ofloxacin. PMID- 16312358 TI - Comparison of the antibacterial activity of lomefloxacin with that of other antibiotic/chemotherapeutic agents. PMID- 16312359 TI - Influence of pH on the postantibiotic effect (PAE) of netilmicin against Staphylococcus aureus and Escherichia coli. PMID- 16312360 TI - Routine susceptibility tests with ticarcillin and tobramycin in combination should be more appropriate than with each antibiotic alone in cystic fibrosis Pseudomonas aeruginosa. PMID- 16312361 TI - Bactericidal activity and post-antibiotic effect in vitro of dactimicin alone and in combination with other drugs. PMID- 16312362 TI - Determination of tetracycline and related compounds by HPLC. PMID- 16312363 TI - Preliminary results on the antibacterial activity of sagamicin towards microorganisms responsible for ocular pathology. PMID- 16312364 TI - In vitro comparative study of seven macrolides against bacterial strains isolated from bronchial aspirates of hospitalized patients. PMID- 16312365 TI - In vitro antibacterial activity of macrolides and other antibiotics against anaerobic bacteria. PMID- 16312366 TI - Effects of erythromycin, flurithromycin and teicoplanin on rat microflora. PMID- 16312367 TI - Bactericidal activity of flucloxacillin and vancomycin in broth, human serum, in the presence of polymorphonuclear leukocytes and on the influence of phagocytosis and intracellular killing against staphylococci. PMID- 16312368 TI - Evaluation of in vitro activity of teicoplanin against gram-positive anaerobic bacteria. PMID- 16312369 TI - Antimicrobial activity of the new macrolide flurithromycin against Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes, Branhamella catarrhalis and Staphylococcus aureus. PMID- 16312370 TI - Activity of teicoplanin alone and in combination. PMID- 16312371 TI - The in vitro activity of the combination of daptomycin and metronidazole against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus spp., and Bacteroides spp. PMID- 16312372 TI - Efficacy of daptomycin compared to teicoplanin in the treatment of Clostridium difficile colitis in hamsters. PMID- 16312373 TI - In vitro bactericidal activity of the glycopeptide compounds vancomycin, teicoplanin and ramoplanin (A-16686/MDL 62,198). PMID- 16312374 TI - A-16686, a new glycopeptide: in vitro evaluation in comparison with other antibiotics. PMID- 16312375 TI - Antibacterial activity on Staphylococcus aureus of original trimethoprim analogues tested alone or in association with sulfamethoxazole. PMID- 16312376 TI - Rifabutine minimum inhibitory concentrations (MICs) for non-tuberculous mycobacteria in liquid medium by a radiometric method. PMID- 16312377 TI - Antibacterial activity of rifaximin against Clostridium difficile, Campylobacter jejunii and Yersinia spp. PMID- 16312378 TI - In vitro antibacterial activity of rifaximin against Campylobacter pylori (Campylobacter pyloridis). PMID- 16312379 TI - Growth and survival of Klebsiella pneumoniae in the presence of pyrithione. PMID- 16312380 TI - Activity in vitro of urine samples from patients treated by nitroxoline against mycoplasmas. PMID- 16312381 TI - A di-thiocarbamido derivative of 2,2'-biphenylenyl-iodonium salt as a potent antibacterial agent. PMID- 16312382 TI - Anti-adherence properties of taurolidine and noxythiolin. PMID- 16312383 TI - The action of imidazole against bacteria. PMID- 16312384 TI - The evolution of azole compounds in antifungal chemotherapy. PMID- 16312385 TI - Action of miconazole at high concentrations against Candida albicans. PMID- 16312386 TI - N, N, N-trimethylamphotericin B methyl ester chloride (DMS-AME): in vitro and in vivo antifungal activity studies in comparison with fungizone. PMID- 16312387 TI - D-mannose in serum as indicator of systemic candidiasis in an experimental model. PMID- 16312388 TI - The effect of growth in the presence of cholic acid on certain cellular constituents of Candida albicans. PMID- 16312389 TI - Experimental vaginal candidosis... and much more! PMID- 16312390 TI - Experimental aspergillosis. PMID- 16312391 TI - Morphobiology of Echinococcus granulosus and therapeutic trials in sheep. PMID- 16312392 TI - Outer membrane alteration and resistance to antimicrobial agents. PMID- 16312393 TI - In vitro studies on the interactions of bacteria with prosthetic devices. PMID- 16312394 TI - Antibiotic susceptibility of slime-producing isolates from orthopedic patients. PMID- 16312395 TI - Influence of gene dosing on antibiotic resistance mediated by inactivating enzymes. PMID- 16312396 TI - Comparative activities in vitro of timentin (ticarcillin/clavulanic acid) and ureidopenicillins against ampicillin-resistant clinical isolates. PMID- 16312397 TI - The effect of subinhibitory concentrations of antibiotics on adherence of slime producing Staphylococcus epidermidis to prosthetic surfaces. PMID- 16312398 TI - Comparative activities of the beta-lactamase inhibitors YTR 830, clavulanic acid and sulbactam combined with ampicillin and piperacillin against beta-lactamase producing anaerobic bacteria. PMID- 16312399 TI - Variations in sensitivity to antibiotics of staphylococci strains isolated from alimentary workers between 1985-1987. PMID- 16312400 TI - Distribution and antibiotic resistance of coagulase-negative staphylococci isolated from human nosocomial infections. PMID- 16312401 TI - Methicillin-resistant staphylococci in surgical and gynecological patients. PMID- 16312402 TI - Incidence and susceptibility of methicillin-resistant staphylococci to 18 antimicrobial agents in Riyadh, Saudi Arabia. PMID- 16312403 TI - Antibiogram of lagos strains of beta-haemolytic streptococci. PMID- 16312404 TI - European Cooperative Study of the prevalence of antibiotic resistance in Haemophilus. PMID- 16312405 TI - Prevalence of antibiotic resistance of Haemophilus influenzae in the F.R.G. PMID- 16312406 TI - Comparative activity of cefotetan against plasmid-encoded aminoglycoside resistant enterobacteria strains. PMID- 16312407 TI - A nationwide survey of Haemophilus influenzae resistance in Poland. PMID- 16312408 TI - Cefuroxime usage and resistance, the Swedish experience. PMID- 16312409 TI - Antimicrobial susceptibilities of Haemophilus influenzae in Hong Kong. PMID- 16312410 TI - Antibiotic resistance, beta-lactamase activity, and plasmid content in strains of Haemophilus and Branhamella isolated from children. PMID- 16312411 TI - Stability of ceftizoxime and its analogues to beta-lactamases. PMID- 16312412 TI - Sensitivity of cefatrizine on clinical isolates of gram-positive, gram-negative cocci, Haemophilus, and Enterobacteriaceae. PMID- 16312413 TI - Characterization of pANM 1, a plasmid coding for sulfonamide resistance in Neisseria meningitidis. PMID- 16312414 TI - Antibiotic sensitivity of urinary isolates in Great Britain during 1983-1985--a multicenter study. PMID- 16312415 TI - Chloramphenicol resistance in clinical isolates of enterobacteria: characterization of chloramphenicol acetyltransferases. PMID- 16312416 TI - Plasmid prevalence in antibiotic resistant Serratia marcescens. PMID- 16312417 TI - Antibiotic resistance transposons on a virulence plasmid from Salmonella wien. PMID- 16312418 TI - Association experiments in antimicrobial susceptibility testing of aerobes and anaerobes to ampicillin and ampicillin plus sulbactam. PMID- 16312419 TI - Mechanism of chloramphenicol and beta-lactam resistance in clinical isolates of Yersinia enterocolitica. PMID- 16312420 TI - Susceptibility profiles of Yersinia spp strains to beta-lactams and other major classes of antimicrobials. PMID- 16312421 TI - Resistance of six frequently isolated bacteria in intensive care unit patients to some cephalosporins and aminoglycosides. PMID- 16312422 TI - Optimal disk susceptibility testing of Staphylococcus epidermidis and aureus against penicillinase-resistant semisynthetic penicillins and cephalosporins. PMID- 16312423 TI - Obtainment of SHV-2 beta-lactamase overproducing mutants from Escherichia coli and Klebsiella pneumoniae clinical isolates. PMID- 16312424 TI - A comparison of the primary sequences of the beta-lactamases SHV-2 and SHV-1. PMID- 16312425 TI - Comparison of hydrolytic properties of SHV-2 and SHV-1 beta-lactamases, particularly concerning third-generation cephalosporins. PMID- 16312426 TI - The effects of clavulanic acid sulbactam on induction of class 1 beta-lactamase. PMID- 16312427 TI - A multifactorial analysis of beta-lactam resistance of 730 Klebsiella spp strains isolated in October, 1985. PMID- 16312428 TI - Prevalence of resistance phenotypes to beta-lactam antibiotics among 4800 isolates of Enterobacteriaceae and distribution of beta-lactamases. PMID- 16312429 TI - In vitro activity of some beta-lactams in combination with dicloxacillin against Mycobacterium fortuitum. PMID- 16312430 TI - Inducing activity of cephalosporins for beta-lactamases in two characterized staphylococcal strains. PMID- 16312431 TI - Effects of beta-lactamase inhibitors on Staphylococcus aureus susceptibility to penicillin G and penicillinase-resistant penicillins. PMID- 16312432 TI - Comparative activity of beta-lactamase inhibitors combined with beta-lactams against antibiotic-resistant enterobacter. PMID- 16312433 TI - Epidemiology of beta-lactamase production and possibilities for the use of augmentin. PMID- 16312434 TI - Activity of amoxycillin/clavulanic acid (augmentin) against beta-lactamase positive S. aureus, H. influenzae and B. catarrhalis in an in vitro kinetic model. PMID- 16312435 TI - Antibiotic susceptibility of Sicilian strains of rickettsia. PMID- 16312436 TI - Timentin (ticarcillin/clavulanic acid): lack of antagonism in an in vitro kinetic model, against bacteria producing inducible class-1 beta-lactamase. PMID- 16312437 TI - Antimicrobial activity of cefixime against antibiotic-resistant Haemophilus influenzae strains. PMID- 16312438 TI - Activity of CM-40874 against Enterobacter resistant to third generation cephalosporins. PMID- 16312439 TI - Comparative in-vitro antibacterial activity of aztreonam against clinical isolates of gram-negative bacteria. PMID- 16312440 TI - The sensitivity to aztreonam and 10 other antibiotics of multiresistant enterobacteria. PMID- 16312441 TI - Differential resistance of hospital Pseudomonas aeruginosa to ceftazidime, imipenem or aztreonam. PMID- 16312442 TI - Sensitivity to beta-lactams and aminoglycosides of clinically important pathogens isolated from children in Poland. PMID- 16312443 TI - Sensitivity of gentamicin-resistant gram-negative bacilli to tobramycin, netilmicin and amikacin as well as synergism with cephalosporins. PMID- 16312444 TI - Aminoglycoside resistance due to alterations of transport mechanisms. PMID- 16312445 TI - In vitro selections of resistant strains by quinolones. PMID- 16312446 TI - New quinolones against coagulase-negative Staphylococcus blood isolates: in vitro sensitivity and slime production. PMID- 16312447 TI - In vitro susceptibility of clinical isolates of staphylococci and Haemophilus influenzae to newer quinolones. PMID- 16312448 TI - In vitro resistance of a 4-quinolone after two years of clinical use. PMID- 16312449 TI - Susceptibility of clinical isolates of Haemophilus to new 4-quinolones. PMID- 16312450 TI - Resistance studies with ofloxacin. PMID- 16312451 TI - Ofloxacin--a possible breakthrough in the treatment of typhoid fever caused by chloramphenicol resistant Salmonella typhi. PMID- 16312452 TI - Biotyping and susceptibility behaviour of pathogens in patients treated with ciprofloxacin. PMID- 16312453 TI - Evaluation of spontaneous resistance to glycopeptide antibiotics in staphylococcal populations. PMID- 16312454 TI - Efficacy of fosfomycin + vancomycin or gentamicin in experimental endocarditis due to methicillin-resistant Staphylococcus aureus (MRSA). PMID- 16312455 TI - Clinical and microbiological evaluation of miocamycin activity against group A beta-haemolytic streptococci in pediatric patients. Three years' incidence of erythromycin-resistant group A streptococci. PMID- 16312456 TI - In vitro susceptibility of Staphylococcus aureus isolated from burn wounds to mupirocin and other antibiotics. PMID- 16312457 TI - Efficacy of mupirocin calcium ointment in the treatment of experimental wound infections caused by methicillin-resistant strains of Staphylococcus aureus. PMID- 16312458 TI - In vitro activity of isepamicin against Pseudomomas aeruginosa and Enterobacter cloacae strains resistant to amikacin: a preliminary report. PMID- 16312459 TI - Plasmid profile as epidemiological marker for Pseudomonas aeruginosa in an intensive care unit. PMID- 16312460 TI - Analysis of cell wall proteins of Candida albicans. PMID- 16312461 TI - Current status and evolution of antimalarial drug resistance (with special reference to chloroquine and related blood schizontocides). PMID- 16312462 TI - Resistance of Mycoplasma pneumoniae to erythromycin: a clinical case with severe pulmonary, cerebral and systemic complications. PMID- 16312463 TI - Host resistance to intracellular parasites and its influence on the efficacy of antibiotic therapy. PMID- 16312464 TI - Evaluation of some immunological parameters during antibiotic therapy. PMID- 16312465 TI - Interaction between antibiotics and non-specific humoral immunity. PMID- 16312466 TI - Effect of quinolones on the production of colony stimulating factors by macrophages. PMID- 16312467 TI - Imipenem influence on the Salmonella typhimurium relationship with BALB/C and CBA mouse macrophages. PMID- 16312468 TI - The effect of clindamycin on human monocytes. PMID- 16312469 TI - Effect of ceftizoxime on interactions between bacteria and professional phagocytic cells. PMID- 16312470 TI - The problem of antibiotic cooperation with the host defence during bacterial infections. PMID- 16312471 TI - Influence of midecamycin acetate, a new macrolide antibiotic, on bactericidal action of human polymorphonuclear leukocytes. PMID- 16312472 TI - Influence of doxycycline, streptomycin, cotrimoxazole and rifamycin on the functions of normal human neutrophils against Brucella melitensis. PMID- 16312473 TI - Increase of phagocytosis and leukocyte intraphagocytic activity of Staphylococcus aureus treated with subinhibitory concentrations of fosfomycin. PMID- 16312474 TI - Polymorphonuclear leukocyte function in children with recurrent infections. PMID- 16312475 TI - P40 modulation of macrophages from different anatomical sites. PMID- 16312476 TI - Endotoxin and macrophage functions. PMID- 16312477 TI - Method for the evaluation of specific bacterial antibodies by Elisa in urine. PMID- 16312478 TI - Specific and nonspecific immunomodulation by wall components of Candida albicans. PMID- 16312479 TI - Lymphokine-activated killer (LAK)-like cells induced in mice after repeated intraperitoneal injections of inactivated Candida albicans. PMID- 16312480 TI - A comparison between interleukin-2- and mannoprotein-activated MHC-unrestricted, killer cells in human peripheral blood mononuclear cells. PMID- 16312481 TI - Cyclosporin A modulates K+ fluxes across the plasma membrane of resting lymphocytes without affecting the intracellular pH and [Ca2+]. PMID- 16312482 TI - Modifications in spleen lymphocyte subsets after cyclophosphamide and 16,16 dimethyl prostaglandin E2 administration in mice. PMID- 16312483 TI - Antimicrobial activity of a macrophage cell line generated in vitro with v-raf/v myc oncogenes. PMID- 16312484 TI - Flow cytometric method for the evaluation of the biological activity of thymic hormones. PMID- 16312485 TI - Differential control of T cell functions by IFN-alpha/beta and IFN-gamma. PMID- 16312486 TI - Association of thymostimulin to antimycobacterial polychemotherapy in pulmonary tuberculosis. PMID- 16312487 TI - Thymopentin (TP 5) in the management of patients with uremia. PMID- 16312488 TI - Improvement of immune response in intravenous drug abusers anti-HIV positive on heroin withdrawal program. PMID- 16312489 TI - Pharmacological modulation of myocardial depressant factor as a mediator of circulatory shock. PMID- 16312490 TI - AD6, a coumarin derivative, and other pharmacological approaches to splanchnic artery occlusion (SAO) shock therapy. PMID- 16312491 TI - Influence of simulated human pharmacokinetics on the efficacy of temocillin against a Klebsiella pneumoniae meningitis infection in the rabbit. PMID- 16312492 TI - Penetration of augmentin and timentin into lymph after simulation of human serum pharmacokinetics in the rabbit. PMID- 16312493 TI - Synergistic antibacterial activity of cefotaxime and the penem HRE 664 in an improved in vitro model simulating serum and tissue pharmacokinetics. PMID- 16312494 TI - Pharmacokinetics of newer antibiotics (enoxacin) in an in vitro placental perfusion model. PMID- 16312495 TI - Pharmacokinetics of newer antibiotics (roxithromycin) in an in vitro placental perfusion model. PMID- 16312496 TI - The importance of pharmacokinetics in respiratory tract infection therapy. PMID- 16312497 TI - Antibacterial activity of piperacillin and gentamicin using a dynamic system simulating in vivo pharmacokinetics. PMID- 16312498 TI - Correlation between pharmacokinetics and therapeutic response in elderly patients. PMID- 16312499 TI - The influence of paracetamol on the gastrointestinal absorption of seven antimicrobials. PMID- 16312500 TI - A comparative pharmacokinetic study between lenampicillin, bacampicillin and amoxycillin. PMID- 16312501 TI - Amoxycillin/clavulanic acid levels in respiratory secretions. PMID- 16312502 TI - The pharmacokinetics of oral beta-lactam prodrugs. PMID- 16312503 TI - Pharmacokinetic and clinical studies on timentin in ear, nose and throat infections. PMID- 16312504 TI - Experimental and clinical evaluation of the biliary pharmacokinetics of ticarcillin plus clavulanic acid (claventin). PMID- 16312505 TI - Pharmacokinetics of YTR830H in healthy human subjects. PMID- 16312506 TI - Pharmacokinetics of YTR830H and piperacillin after intravenous coadministration. PMID- 16312507 TI - Direct evaluation of mezlocillin concentrations in bronchial tissue. PMID- 16312508 TI - Pharmacokinetics and metabolism of FCE 22101, a new penem antibiotic. PMID- 16312509 TI - Pharmacokinetics and clinical tolerability of FCE 22101, a new penem antibiotic, in healthy volunteers after a single i.m. administration. PMID- 16312510 TI - Tissue penetration following orally administered cefuroxime axetil. PMID- 16312511 TI - Pharmacokinetics in the elderly with reference to the oral prodrug cefuroxime axetil. PMID- 16312512 TI - Clinical and pharmacokinetic study of cefotetan in biliary tract infections: preliminary report. PMID- 16312513 TI - Concentrations of ceftriaxone in plasma and in pus in patients with soft tissue abscesses. PMID- 16312514 TI - Comparative pharmacokinetics of ceftriaxone, cefmetazole and moxalactam during abdominal surgery. PMID- 16312515 TI - Prostatic concentrations of beta-lactam antibiotics. PMID- 16312516 TI - Cefotaxime and ceftriaxone levels in pancreas tissue and pancreatic juice. PMID- 16312517 TI - Concentrations of ceftriaxone in expressed prostatic fluid of males with chronic prostatitis. PMID- 16312518 TI - Pharmacokinetic and clinical studies of cefixime: a worldwide review. PMID- 16312519 TI - Ceftazidime in patients with impaired renal function. Studies on pharmacokinetics and nephrotoxicity. PMID- 16312520 TI - The pharmacokinetics of cefixime in patients with renal insufficiency undergoing hemofiltration. PMID- 16312521 TI - Antibiotic levels in the fluid in pancreatic pseudocysts aspirated by ultrasound- guided percutaneous puncture. PMID- 16312522 TI - Effect of method of antibiotic administration on pleural exudate penetration: a study of aztreonam given by rapid injection and constant infusion. PMID- 16312523 TI - Aztreonam concentrations in colonic tissue in patients undergoing abdominal surgery. PMID- 16312524 TI - Bioavailability and pharmacokinetics of enoxacin. PMID- 16312525 TI - Bone and soft tissue concentrations of aztreonam after intravenous administration. PMID- 16312526 TI - Clinical pharmacokinetics of enoxacin in subjects with renal or hepatic impairment. PMID- 16312527 TI - Penetration of ciprofloxacin into human lung tissue after oral and i.v. dosing. PMID- 16312528 TI - Ciprofloxacin concentration in bone tissue. PMID- 16312529 TI - Antimicrobial impregnation of cartilage implants with ciprofloxacin. PMID- 16312530 TI - Ciprofloxacin--clincal and pharmacokinetic studies in ear, nose and throat infections. PMID- 16312531 TI - Fleroxacin pharmacokinetics in healthy volunteers and impaired renal function. PMID- 16312532 TI - Bronchopulmonary penetration of ciprofloxacin. PMID- 16312533 TI - Concentration of ciprofloxacin and metronidazole in human tissues during abdominal surgery. PMID- 16312534 TI - Ciprofloxacin and oxociprofloxacin pharmacokinetics in patients on haemodialysis. PMID- 16312535 TI - Ciprofloxacin skin titers in old age. PMID- 16312536 TI - Penetration of ciprofloxacin into the mucosa of the human maxillary sinus. PMID- 16312537 TI - Pharmacokinetics of fleroxacin determined by biological method. PMID- 16312538 TI - Pharmacokinetics of rufloxacin (MF-934) in normal human volunteers after single oral administration. PMID- 16312539 TI - A study of ciprofloxacin kinetics in human muscle and gut tissues. PMID- 16312540 TI - Clinical and pharmacokinetic evaluation of josamycin in the treatment of inflammatory acne. PMID- 16312541 TI - Effect of dose on macrolide distribution in saliva and periodontal tissues of young patients. PMID- 16312542 TI - Pharmacokinetic data and therapeutic efficacy of miocamycin: serum, urinary and prostatic levels. Therapeutic aspects in acute urethral syndrome and non gonococcal urethritis. PMID- 16312543 TI - Miocamycin in chlamydia trachomatis infections in neonatology: clinical results, pharmacokinetics and microbiological evaluation. PMID- 16312544 TI - Some features of miocamycin in the gynecological field: pharmacokinetic data using preoperative administration and therapeutic aspects against chlamydial infections. PMID- 16312545 TI - Roxithromycin penetration into alveolar bone and gum. PMID- 16312546 TI - Concentrations of vancomycin in human bone. PMID- 16312547 TI - Pharmacological profile of teicoplanin. PMID- 16312548 TI - Pharmacokinetics and tolerance of a new film-coated tablet of sodium fusidate administered as a single oral dose to healthy volunteers. PMID- 16312549 TI - The pharmacokinetics of sulfapyridine and its N4-acetyl-, and hydroxy metabolites in man. PMID- 16312550 TI - Impact of lenampicillin on the fecal flora of healthy volunteers. PMID- 16312551 TI - Antimicrobial agents and neuromuscular blockade. PMID- 16312552 TI - Effect of cefotaxime on antibody-producing cells of mouse spleen. PMID- 16312553 TI - Impact of ciprofloxacin and cotrimoxazole on bone marrow growth as measured by CFU-GM and BFU-E assays. PMID- 16312554 TI - No influence of intravenous ciprofloxacin on histamine release, complement activation, and blood clotting system. PMID- 16312555 TI - Comparative epileptogenic activity of various cephalosporins in rats. PMID- 16312556 TI - Effect of ofloxacin on human colonic microflora. PMID- 16312557 TI - Inhibition of vas deferens contractility caused by aminoglycoside antibiotics. PMID- 16312558 TI - Effect of oral ciprofloxacin on fecal bacteria in elderly patients. PMID- 16312559 TI - Interaction of aminoglycoside antibiotics with calcium channel blockers at the neuromuscular junctions. PMID- 16312560 TI - Interaction of aminoglycoside antibiotics with H2-blockers and calcium channel blockers in isolated smooth muscles of rat aorta. PMID- 16312561 TI - Interaction of aminoglycoside antibiotics with steroidnon-depolarizing muscle relaxant agents. PMID- 16312562 TI - Evaluation of the therapeutic efficacy and tolerability of teicoplanin: European multicenter study and personal experience. PMID- 16312563 TI - Photo-sensitization by sulfamethoxazole-trimethoprim through the skin: a case report. PMID- 16312564 TI - Side effects of treatment with antimony salts. PMID- 16312565 TI - Impact of Lactobacillus acidophilus administration on the intestinal microflora after clindamycin treatment. PMID- 16312566 TI - Acute abdominal pain during therapy with glucantim: splenic infarct or excessive splenic enlargement? PMID- 16312567 TI - Etiology, epidemiology and chemotherapy of nosocomial infections: medico-legal aspects. PMID- 16312568 TI - Imipenem in the treatment of infections at different sites. PMID- 16312569 TI - A clinical study of imipenem/cilastatin in the treatment of generalized and localized infections. PMID- 16312570 TI - A review of clinical studies on lenampicillin in Japan. PMID- 16312571 TI - Clinical evaluation of imipenemcilastatin in the therapy of severe infections in high risk patients. PMID- 16312572 TI - A world-wide summary of clinical experience with augmentin. PMID- 16312573 TI - Efficacy and tolerance of cefuroxime axetil: an overview of the Italian experience. PMID- 16312574 TI - Clinical efficacy of cefotaxime in two different periods. PMID- 16312575 TI - Clinical experience with lower doses of cefotaxime in hospitalized patients with serious infections. PMID- 16312576 TI - A study of cefotaxime versus netilmicin versus netilmicin and cefotaxime in serious infections. PMID- 16312577 TI - Ambulatory treatment with intramuscular ceftriaxone. PMID- 16312578 TI - Further observations of the role of timentin in clinical practice. PMID- 16312579 TI - A clinical trial of ceftriaxone in different bacterial infections. PMID- 16312580 TI - Lack of correlation between in vitro susceptibility data of anaerobes to ceftizoxime and clinical response. PMID- 16312581 TI - Clinical impact of in vitro data with newer quinolones. PMID- 16312582 TI - Are the quinolones realising their clinical potential? PMID- 16312583 TI - Post-marketing surveillance with ceftizoxime in 5081 elderly patients. PMID- 16312584 TI - Pefloxacin in the treatment of acute severe infections in hospitalized patients. Clinical and bacteriological evaluation. PMID- 16312585 TI - A review of the therapeutic uses of ofloxacin. PMID- 16312586 TI - Serious, life-threatening infections: results with pefloxacin. PMID- 16312587 TI - Ciprofloxacin in the treatment of infections at different sites. PMID- 16312588 TI - Follow-up therapy with ciprofloxacin in severe infection, an alternative to parenteral treatment. PMID- 16312589 TI - Treatment of pediatric bacterial meningitis with a 7-day regimen of once-daily ceftriaxone injections. Multicentre study carried out in non-university pediatric departments in the French and Italian-speaking regions of Switzerland. PMID- 16312590 TI - Efficacy of ofloxacin taken orally in 19 severe infections. PMID- 16312591 TI - Ceftriaxone vs. piperacillin in patients with bacterial meningitis. PMID- 16312592 TI - First data about imipenem in the treatment of pediatric infections. PMID- 16312593 TI - Aztreonam treatment of neonatal sepsis. PMID- 16312594 TI - The usefulness of erythromycin in paediatrics. PMID- 16312595 TI - Cefixime versus amoxicillin in the treatment of acute otitis media in infants and children. PMID- 16312596 TI - Study on causative microbes and antibiotics sensitivity in infants suffering from urinary tract infections with congenital malformations. PMID- 16312597 TI - The epidemiology of Haemophilus influenzae type b disease. PMID- 16312598 TI - The control of hospital infections in a local health unit. PMID- 16312599 TI - Contribution to the monitoring of antibiotic therapy by the bactericidal serum activity test. PMID- 16312600 TI - Staphylococcus aureus isolates from blood cultures. PMID- 16312601 TI - Revival of gram-positive bacteria as superficial infectious etiological agents. PMID- 16312602 TI - Antimicrobial combinations in the treatment of gram-positive infections. PMID- 16312603 TI - The international experience with teicoplanin in the treatment of skin and soft tissue infections. PMID- 16312604 TI - Teicoplanin treatment of Corynebacterium JK infection and colonization in an immunocompromised patient before and after marrow transplantation. PMID- 16312605 TI - Evaluation of teicoplanin's therapeutic efficacy in immunocompromised patients with staphylococcal sepsis. PMID- 16312606 TI - Teicoplanin in the therapy of gram-positive infections: an open non-randomized study. PMID- 16312607 TI - Ciprofloxacin treatment of acute infections in immunocompromised host. PMID- 16312608 TI - Anaerobic bacteremia in bone marrow recipients. PMID- 16312609 TI - Effects of norfloxacin or pefloxacin on the oral and intestinal microflora of bone marrow transplant patients. PMID- 16312610 TI - Microbial and non-microbial infections in renal transplant recipients: ten-year experience. PMID- 16312611 TI - Microbial contamination and wound infections in open fractures. PMID- 16312612 TI - Spectrum and sensitivity determination of causative pathogens in post-traumatic osteomyelitis. PMID- 16312613 TI - The treatment of orthopedic infections with ceftriaxone. PMID- 16312614 TI - Oral enoxacin versus conventional intravenous antimicrobial therapy for chronic osteomyelitis. PMID- 16312615 TI - Ofloxacin in open fractures. PMID- 16312616 TI - Experience in infective endocarditis. PMID- 16312617 TI - Occurrence of prosthetic valve endocarditis due to oxacillin-sensitive and oxacillin- resistant. Staphylococci, and the possibility 5 of therapy. PMID- 16312618 TI - Pathogenesis and laboratory diagnosis of oral cavity infections. PMID- 16312619 TI - Treatment of severe odontogenic infections with amoxicillin/clavulanic acid. PMID- 16312620 TI - The activity of ofloxacin compared to spiramycin in oral surgery. PMID- 16312621 TI - Comparison of two bacampicillin therapeutic regimens in infections of middle ear, pharynx and paranasal sinuses. PMID- 16312622 TI - Study of the efficacy of amoxicillin plus clavulanic acid in the treatment of ear, nose and throat infections. PMID- 16312623 TI - The local application of azlocillin in otitis externa. PMID- 16312624 TI - Cefotiam in ear, nose, and throat infectious pathology. PMID- 16312625 TI - Low-dose ciprofloxacin in the treatment of bacterial pharyngo-tonsillitis. PMID- 16312626 TI - Therapeutic considerations for Haemophilus influenzae infections. PMID- 16312627 TI - Imipenem/cilastatin as monotherapy in the treatment of acute or chronic lung infections: clinical efficacy and safety. PMID- 16312628 TI - Cefuroxime axetil: management of adult sinusitis in the clinic. PMID- 16312629 TI - A comparison of cefuroxime axetil and amoxycillin in the treatment of lower respiratory tract infections. PMID- 16312630 TI - Treatment of respiratory tract infections with augmentin. PMID- 16312631 TI - A comparison of the safety and efficacy of cefuroxime axetil and cefaclor in outpatient management of community-acquired lower respiratory tract infections. PMID- 16312632 TI - Cefuroxime axetil in the treatment of acute bronchitis in UK general practice. PMID- 16312633 TI - A multicentre trial with cefotiam in respiratory tract infections. PMID- 16312634 TI - Cefixime once-a-day in the treatment of lower respiratory tract infections in adults. PMID- 16312635 TI - Cefetamet pivoxil (Ro 15-8075) and amoxicillin in the treatment of bacterial exacerbations of chronic bronchial disease--preliminary results. PMID- 16312636 TI - Cefixime: comparative clinical trial on the treatment of upper respiratory tract infection and otitis in adults. PMID- 16312637 TI - Efficacy and safety in the oral treatment of purulent chest disease and pneumonia with cefixime, ofloxacin, and ciprofloxacin. PMID- 16312638 TI - Ofloxacin versus amoxicillin in the treatment of lower respiratory tract infections. A multicenter prospective clinical trial. PMID- 16312639 TI - Cefixime: comparative clinical trial on the treatment of lower respiratory tract infection in adults. PMID- 16312640 TI - Effect of combination therapy with ciprofloxacin and azlocillin in intensive care patients with pneumonia and/or sepsis. PMID- 16312641 TI - Enoxacin: clinical results in lower respiratory tract infections. PMID- 16312642 TI - Comparative clinical study of clofoctol versus josamycin in respiratory tract infections. PMID- 16312643 TI - Treatment of bronchopulmonary infections by ciprofloxacin in an intensive care unit. Correlation between serum and bronchial concentrations and bacteriological results. PMID- 16312644 TI - Clinical efficacy of roxithromycin in respiratory tract infections. PMID- 16312645 TI - An open, comparative study on trimethoprim and doxycycline in the treatment of acute bronchitis. PMID- 16312646 TI - The biology of human mycoplasmas: a brief review. PMID- 16312647 TI - Some methodological problems in the control of human mycoplasma infections. PMID- 16312648 TI - In vitro activity of eleven newer antibiotics against bacterial enteropathogens associated with acute fevers and diarrheal disease in Egypt. PMID- 16312649 TI - The use of quinolones in the treatment of acute bacterial diarrhea: a comparative therapeutic trial. PMID- 16312650 TI - Intensive care unit environmental contamination and positive bronchial cultures in critically ill patients. Is there a relationship? PMID- 16312651 TI - Critical role of bacterial mixtures in severe surgical infections. PMID- 16312652 TI - Our management of acute complicated appendicitis in children. PMID- 16312653 TI - Comparative study of mezlocillin in combination with metronidazole in patients with perforation of the inflamed appendix. PMID- 16312654 TI - Piperacillin vs gentamicin-clindamycin combination in intra-abdominal infections. PMID- 16312655 TI - Cephalosporin therapy in intraabdominal infection: comparative studies of cefotetan, latamoxef and cefoxitin. PMID- 16312656 TI - The antibiotic regimen for patients with suppurative peritonitis during peritoneal lavage program. PMID- 16312657 TI - Cefotetan versus cefoxitin in the treatment of patients with biliary sepsis assessed by a biliary sepsis score. PMID- 16312658 TI - Combined cephalosporin aminoglycoside therapy in gram-negative sepsis. The choice of cephalosporin. PMID- 16312659 TI - IV/PO ciprofloxacin in treatment of surgical infections. PMID- 16312660 TI - Clinical trial of norfloxacin in the treatment of bacterial enteritis. PMID- 16312661 TI - Septicaemia in chronic renal failure. PMID- 16312662 TI - Colloidal bismuth subcitrate (DE-NOL) and tinidazole healed duodenal ulceration with a low relapse rate due to elimination of Campylobacter pylori. PMID- 16312663 TI - New possibilities in the treatment of urinary tract infections. PMID- 16312664 TI - Treatment and prevention of bacteriuria in patients on median-term transurethral catheterization. PMID- 16312665 TI - Evaluation of the efficacy and safety of imipenem/cilastatin in the treatment of complicated urinary tract infections. PMID- 16312666 TI - Cefotaxime in a single daily dose in lower urinary tract infections. PMID- 16312667 TI - Treatment of uncomplicated urinary infections with a single dose of Ro 15-8075 cefetamet pivoxyl. PMID- 16312668 TI - The use of enoxacin in the treatment of urinary disease: a critical review of the Italian and international literature. PMID- 16312669 TI - In vitro and clinical evaluation of ofloxacin in urinary tract infection and enteric fever. PMID- 16312670 TI - Ofloxacin in the treatment of genito-urinary tract infections. PMID- 16312671 TI - Experience with ofloxacin for short and long-term treatment of urinary tract infections in renal transplant recipients. PMID- 16312672 TI - Ciprofloxacin versus amoxycillin/clavulanic acid in the treatment of urinary tract infections in general practice. PMID- 16312673 TI - Tobramycin once-a-day vs cefoperazone twice-a-day in the therapy of urinary tract infections. PMID- 16312674 TI - Amikacin--single dose treatment for uncomplicated urinary tract infections in children. PMID- 16312675 TI - Acute, uncomplicated, lower urinary tract infections: the role of single-dose fleroxacin. PMID- 16312676 TI - Single dose of netilmicin in urinary tract infection. PMID- 16312677 TI - Efficacy of long-term therapy with norfloxacin in chronic bacterial prostatitis. PMID- 16312678 TI - Clinical efficacy of ofloxacin (tarivid) in patients with chronic bacterial prostatitis: preliminary results. PMID- 16312679 TI - Evaluation of mezlocillin in treatment of acute and chronic prostatitis. PMID- 16312680 TI - Clinical experience in gynecology on the enhancement of specific therapy using alkyl amino-ethylglycine, an amphoteric surfactant with amino acid structure. PMID- 16312681 TI - Imipenem/cilastatin in a group of obstetrics/gynecological surgical patients. PMID- 16312682 TI - Puerperal infections: experiences with imipenem/cilastatin. PMID- 16312683 TI - Cefotetan administration in the treatment of post-cesarean endometritis. PMID- 16312684 TI - The treatment of gynaecological and intra-abdominal infections: a comparative study of cefotetan versus netilmicin plus clindamycin. PMID- 16312685 TI - Comparison of ceftizoxime versus combination therapy in the treatment of pelvic inflammatory disease (PID). PMID- 16312686 TI - A comparison of ceftriaxone (1 g single dose) versus cefotaxime (three 1 g doses) for gynecologic and obstetric infections. PMID- 16312687 TI - Enoxacin in urinary tract infections associated with gynecological diseases. PMID- 16312688 TI - Efficacy and safety of ciprofloxacin in gynaecological infections. PMID- 16312689 TI - Treatment of soft tissue infections in gynecology by comparison 3 vs 7 day therapy with ciprofloxacin. PMID- 16312690 TI - A review of augmentin treatment regimes in gonorrhoea due to penicillinase producing Neisseria gonorrhoeae: 1981-1987. PMID- 16312691 TI - A comparative trial of cefuroxime axetil and ampicillin plus probenecid in the treatment of acute gonococcal infection. PMID- 16312692 TI - Outpatient treatment of gonorrhea with ceftriaxone. PMID- 16312693 TI - Clinical efficacy and safety of oral ciprofloxacin in pelvic inflammatory disease. PMID- 16312694 TI - Evaluation of an IgM antibody capture enzyme immunoassay Captia Syphilis-M in treated and untreated syphilis. PMID- 16312695 TI - Treatment of Chlamydia trachomatis infections. PMID- 16312696 TI - Chlamydial infections in children over 1 year of age: experience with josamycin. PMID- 16312697 TI - Frequency of Chlamydia trachomatis, Mycoplasma hominis and Ureaplasma urealyticum infections in pregnant women. PMID- 16312698 TI - Josamycin in the treatment of chlamydial genital infections in infertile women. PMID- 16312699 TI - Josamycin in the treatment of Chlamydia trachomatis cervicitis. PMID- 16312700 TI - Treatment of Chlamydia trachomatis endometritis with josamycin. PMID- 16312701 TI - Role of Chlamydia trachomatis in chronic prostatitis: diagnostic investigation and therapeutic experience with miocamycin. PMID- 16312702 TI - Therapeutic considerations about psittacosis: remarks on a case of family psittacosis. PMID- 16312703 TI - Ceftriaxone--An effective treatment of late Lyme borreliosis. PMID- 16312704 TI - Ofloxacin in the treatment of boutonneuse fever. A preliminary report. PMID- 16312705 TI - Josamycin versus minocycline in the treatment of papulopustular acne. PMID- 16312706 TI - Nosography and therapy of opportunistic infections of the central nervous system in AIDS patients. PMID- 16312707 TI - Treatment of invasive fungal infections in immunocompromised patients with fluconazole. PMID- 16312708 TI - Duration of parenteral hyperalimentation and candidemia in severely ill hospitalized patients. PMID- 16312709 TI - Cryptococcal antigen survey among racing pigeon subjects, patients with cryptococcosis versus pythiosis, histoplasmosis and penicilliosis: serotyping of Cryptococcus. PMID- 16312710 TI - Zygomycetes and Pythium spp., an aquatic phycomycetes isolated from patients and their host defense mechanism. PMID- 16312711 TI - Efficacy and tolerance of local treatment of acute vulvovaginal candidiasis with 4 different single doses (10, 25, 50 and 100 mg) of Ro 14-4767 versus a 7-day treatment with miconazole. PMID- 16312712 TI - New perspectives in the chemotherapy of toxoplasmosis. PMID- 16312713 TI - Epidemiology of human hydatidosis from E. granulosus. PMID- 16312714 TI - Role of ultrasound in the diagnosis and treatment of hydatid liver cysts. PMID- 16312715 TI - Ultrasound examination in patients with hydatid cysts during albendazole treatment. PMID- 16312716 TI - Albendazole in the treatment of hydatid disease. PMID- 16312717 TI - The treatment of giardiasis with albendazole. PMID- 16312718 TI - Chemotherapy of toxoplasmic encephalitis in AIDS patients. PMID- 16312719 TI - Biotechnological applications for the diagnosis and prevention of parasitic diseases. PMID- 16312720 TI - Up-date on the treatment of ocular toxoplasmosis. PMID- 16312721 TI - Visceral leishmaniasis therapy. PMID- 16312722 TI - Our experiences in visceral leishmaniasis therapy. PMID- 16312723 TI - Chemotherapy of human trypanosomiases. PMID- 16312724 TI - Endogenous risk factors of nosocomial infections and degree of prevention. PMID- 16312725 TI - Anti-infective prophylaxis with single i.v. infusion of ceftizoxime in various surgical procedures. A multicentre study. PMID- 16312726 TI - Incidence of oral bacteremia and antimicrobial prophylaxis. PMID- 16312727 TI - Practical regimens for prophylaxis and early treatment of infections in high risk operations. PMID- 16312728 TI - Prophylaxis of dental bacteremia. PMID- 16312729 TI - Antibiotic prophylaxis in gastric, biliary and colonic surgery. PMID- 16312730 TI - Cefotetan short-term prophylaxis in abdominal surgery. Preliminary report. PMID- 16312731 TI - A trial of intra-incisional versus intra-venous timentin as prophylaxis in abdominal surgery. PMID- 16312732 TI - Antibiotic prophylaxis in high-risk biliary tract surgery: a comparison of mezlocillin and cefotetan. Preliminary results. PMID- 16312733 TI - Single-dose prophylaxis with cefotetan in elective abdominal surgery. A controlled trial. PMID- 16312734 TI - A comparison of cefotetan and cefoxitin as prophylaxis in high risk biliary surgery. PMID- 16312735 TI - Experiences with cefotaxime in the prophylaxis of biliary tract surgery. PMID- 16312736 TI - Microbiological intraoperative results in short-term prophylaxis (aztreonam or gentamicin) in abdominal surgery. PMID- 16312737 TI - Has enterococcus to be regarded as pathogenic in biliary surgery? Comments on antibiotic prophylaxis using only one preoperative dose of ceftriaxone. PMID- 16312738 TI - Aztreonam versus gentamicin in short-term prophylaxis in biliary and gastric surgery. PMID- 16312739 TI - Clinical and antibacterial efficacy of ciprofloxacin after single-shot prophylaxis during cholecystectomy. PMID- 16312740 TI - Antibiotic prophylaxis in colorectal surgery. PMID- 16312741 TI - Single-dose prophylaxis in elective colorectal surgery: a prospective, randomized trial with piperacillin or latamoxef. PMID- 16312742 TI - A prospective comparative study of cefotetan versus cefoxitin against infection in elective colorectal surgery. PMID- 16312743 TI - The pharmacokinetic properties of cefotetan and its relevance for prophylaxis in elective colorectal surgery. PMID- 16312744 TI - Experiences with ceftriaxone (rocephin) in perioperative antibiotic prophylaxis for elective colon surgery. PMID- 16312745 TI - Pre-operative single-dose broad spectrum prophylaxis with ceftriaxone and ornidazole in elective colon surgery. PMID- 16312746 TI - Colorectal surgery: short-term prophylaxis with aztreonam plus clindamycin versus gentamicin plus clindamycin. PMID- 16312747 TI - Cefoperazone or cefoperazone-sulbactam as perioperative antibiotic prophylaxis in transurethral surgery. PMID- 16312748 TI - Perioperative prophylaxis in colorectal surgery: is a single shot with 200 mg ciprofloxacin sufficient to maintain therapeutic levels in gut tissues? PMID- 16312749 TI - Prevention of urinary tract infections in elderly diabetic patients with cefotetan. PMID- 16312750 TI - Ceftriaxone as prophylaxis in endourology and extracorporal shockwave lithotripsy (ESWL). AB - The high risk of infection in endourological surgery is caused by manipulation of the occluded and frequently infected upper urinary tract with instruments. The results show a significant decrease of infection by antibiotic prophylasix with ceftriaxone. However prophylaxis is not necessary in ESWL. PMID- 16312751 TI - Antibiotic prophylaxis with aztreonam in patients with kidney stone disease submitted to extracorporeal shock wave lithotripsy. PMID- 16312752 TI - Single-dose and short-course netilmicin prophylaxis in urologic surgery: pharmacokinetic and clinical evaluation. PMID- 16312753 TI - Antibiotic prophylaxis in emergency and elective cesarean section. PMID- 16312754 TI - Piperacillin in antibiotic prophylaxis for gynecological surgery. PMID- 16312755 TI - Piperacillin in antibiotic prophylaxis: a single-dose administration for cesarean section. PMID- 16312756 TI - Piperacillin: as a single-dose prophylactic agent in hysterectomies. PMID- 16312757 TI - Antimicrobial prophylaxis in non-infected patients undergoing abdominal or vaginal hysterectomy or cesarean section. Comparative efficacy of a single preoperative dose of ceftriaxone and of multiple doses of combined amoxicillin plus metronidazole and of amoxicillin alone. PMID- 16312758 TI - A comparison of mezlocillin and cefotetan in cesarean section prophylaxis: a prospective, randomized study. Preliminary results. PMID- 16312759 TI - Prophylactic mezlocillin in vaginal hysterectomy: a prospective, randomized study. PMID- 16312760 TI - Mezlocillin versus cefotetan in abdominal hysterectomy prophylaxis: a prospective randomized study. Preliminary results. PMID- 16312761 TI - Single shot augmentin as prophylaxis in hysterectomy: preliminary results of a prospective randomised comparative study. PMID- 16312762 TI - The efficacy of ceftriaxone administered for prophylaxis of postoperative infection and infectious diseases in obstetrics and gynecology. PMID- 16312763 TI - Cefotetan and ceftriaxone for single-dose prophylaxis in cesarean section. PMID- 16312764 TI - Ciprofloxacin prophylaxis for vaginal and abdominal hysterectomy--preliminary results. PMID- 16312765 TI - Perioperative antibacterial prophylaxis in bone surgery: sufficiently high tissue levels with ceftriaxone (rocephin) 24 hours after a single dose of 2 g. PMID- 16312766 TI - Comparative study of ceftriaxone versus cefamandole for pre-operative prophylaxis of infections in patients undergoing cesarean section or vaginal hysterectomy. PMID- 16312767 TI - Ceftriaxone versus cefazolin for prophylaxis in orthopedic surgery. PMID- 16312768 TI - Comparative clinical and pharmacokinetic aspects of cefotetan versus cefoxitin plus metronidazole in vaginal hysterectomy. PMID- 16312769 TI - Comparative study of ceftriaxone (rocephin) and cefazolin (kefzol) in perioperative prophylaxis in operations of open fractures. PMID- 16312770 TI - Prophylactic activity of amoxicillin/clavulanic acid and cefoxitin in vascular surgery--a randomized clinical study. PMID- 16312771 TI - Prophylactic mezlocillin-netilmicin combination in pacemaker surgery: a prospective, randomized study. PMID- 16312772 TI - Perioperative prophylaxis with ciprofloxacin in vascular surgery. PMID- 16312773 TI - Perioperative antibiotic prophylaxis in patients undergoing graft vascular surgery. PMID- 16312774 TI - Prophylaxis and therapy by mezlocillin in pediatric surgery. PMID- 16312775 TI - Biotechnology for virus vaccine design, production, and control. PMID- 16312776 TI - Triterpenes potentiate antiviral activity of prostaglandin A. PMID- 16312777 TI - Inhibition of the replication of DNA viruses by phosphonylmethoxyalkyl-purines and -pyrimidines. PMID- 16312778 TI - High beta2 microglobulin serum levels in human immunodeficiency virus infected intravenous drug addicts. PMID- 16312779 TI - Influenza virus prophylaxis. PMID- 16312780 TI - Antiviral activity of natural 3-methoxyflavones. PMID- 16312781 TI - Antiviral activity of synthetic 3-methoxyflavones. PMID- 16312782 TI - The role of thymopentin in hemodialyzed hepatitis B vaccine non-responders. PMID- 16312783 TI - Interferon in chronic active hepatitis B: preliminary results. PMID- 16312784 TI - Interferon treatment of anti-HBe positive and HBV-DNA positive chronic hepatitis type B. PMID- 16312785 TI - Response to interferon treatment with or without steroids in 20 children with chronic hepatitis B. PMID- 16312786 TI - Treatment of chronic hepatitis B with human lymphoblastoid interferon: results of a controlled trial. PMID- 16312787 TI - Clinical experience with a yeast-derived hepatitis B vaccine. PMID- 16312788 TI - Delta infection in Portuguese drug addicts. PMID- 16312789 TI - Evaluation of a solid-phase fluorescence immunoassay (FIAX) for detection of human IgG antibodies to cytomegalovirus. PMID- 16312790 TI - Cytomegalovirus acute hepatitis: considerations about acyclovir treatment. PMID- 16312791 TI - Rapid diagnosis of herpes virus infections: a short review. PMID- 16312792 TI - A novel pharmacological approach to herpes virus infections. PMID- 16312793 TI - Three severe relapsing herpes labialis treated with thymopentine. PMID- 16312794 TI - In vitro sensitivity of clinical isolates and drug-resistant mutants of herpes simplex virus against seven antiviral drugs. PMID- 16312795 TI - Treatment of chronic hepatitis B with a combination of acyclovir and alpha lymphoblastoid interferon. PMID- 16312796 TI - Immunohistochemical diagnosis of herpetic gingivostomatitis and its treatment with acyclovir. PMID- 16312797 TI - Efficacy of acyclovir in the treatment of recurring herpes labialis, genitalis and acute herpes zoster. PMID- 16312798 TI - Treatment of papillomavirus infections in teenagers. PMID- 16312799 TI - Papillomavirus infections in teenagers: a clinical and epidemiological study. PMID- 16312800 TI - Verruca plana juvenilis (plane wart) treated with topical application of acyclovir. PMID- 16312801 TI - Estrogen receptor binding and alkylating properties of A-ring aromatic 17,20- and 20,21-epoxy-norpregnanediols: potential target selective antitumour agents. PMID- 16312802 TI - Linkage analysis of the multidrug-resistant gene family by somatic cell genetics study. PMID- 16312803 TI - Pharmacokinetic and cell-biological investigation of prednimustine compared to its components chlorambucil and prednisolone. PMID- 16312804 TI - Multiply drug-resistant human CCRF-CEM T-lymphoblastoid cells have decreased amount of a 90 KDa cell surface protein. PMID- 16312805 TI - M-VAC regimen in murine bladder tumors. PMID- 16312806 TI - Amiodarone cytotoxicity on tumour cell cultures. PMID- 16312807 TI - DNA ligases are specifically inhibited by anthracyclines with a free 3'-amino group. PMID- 16312808 TI - Biology and mode of action of pure antioestrogens. PMID- 16312809 TI - Primary screening of cytotoxic activity of 6-purinyl-N-(2-chlordethyl) thiocarbamate. PMID- 16312810 TI - Tumoristatic action of tamoxifen: experimental evidence for clinical application in stage I and II breast cancer. PMID- 16312811 TI - Chemistry and biological activity of hematoporphyrin derivative (HPD) and photofrin II on human cancerous cells. PMID- 16312812 TI - Possible role of mitochondrial components in adriamycin-induced cytotoxicity of human leukemia cells. PMID- 16312813 TI - Modifications in morphology and size distribution induced by adriamycin in cultured leukemia cells. PMID- 16312814 TI - Effect of hemin-induced differentiation on chemosensitivity of K 562 leukemia cells. PMID- 16312815 TI - Chemosensitivity of drug-sensitive and drug-resistant P388 leukemia cell lines after hyperthermic treatment. PMID- 16312816 TI - The multipotential hematopoietic growth factors, bryostatins, inhibit human leukemia cell lines in vitro. PMID- 16312817 TI - Increase of NK activity of irradiated mononuclear cells induced by interferon in vitro. PMID- 16312818 TI - Combined treatment with thymosin alpha 1 and interferon enhances NK activity in immunosuppressed tumor bearing mice. PMID- 16312819 TI - Tumor necrosis factor-alpha secretion by splenocytes of Candida albicans infected mice. PMID- 16312820 TI - Timunox increases resistance to the challenge with a low number of cells from Yoshida ascites tumor. PMID- 16312821 TI - Regulation of cell proliferation by inducible-secreted proteins in growth factor treated EL2 rat fibroblasts. PMID- 16312822 TI - Tumor lysis of a glioblastoma by Clostridium oncolyticum. PMID- 16312823 TI - Resistance of human malignant intracranial tumors in vitro: potential guidelines to clinical therapy? PMID- 16312824 TI - Brain tissue concentration of mitoxantrone in the rat in relation to in vitro chemosensitivity of malignant human intracranial tumors--preliminary results. PMID- 16312825 TI - Plasma levels of ACNU in humans and tissue concentrations in the rat brain. PMID- 16312826 TI - Increased dosage of epirubicin and mitomycin-C for the treatment of metastatic adenocarcinoma of unknown primary site. PMID- 16312827 TI - Mastectomy followed by cytoreductive chemotherapy in locally advanced breast cancer. PMID- 16312828 TI - Second and third line chemotherapy in advanced breast cancer. PMID- 16312829 TI - Mitoxantrone and prednimustine combination chemotherapy in breast cancer. PMID- 16312830 TI - Arterial regional chemotherapy of advanced breast cancer. PMID- 16312831 TI - Tamoxifen versus tamoxifen + mitoxantrone treatment in metastatic breast cancer. PMID- 16312832 TI - Interleukin-2 receptors in patients with lung carcinoma metastasis undergoing treatment with disodium-clodronate. Preliminary research. PMID- 16312833 TI - Mitoxantrone-cyclophosphamide-fluorouracil-combination chemotherapy in metastatic breast cancer. 3-Year-follow-up. PMID- 16312834 TI - A randomized trial on adjuvant treatment of stage I lung cancer with retinol palmitate: preliminary data at three years. PMID- 16312835 TI - Cis-platin, vindesine, doxorubicin and cyclophosphamide in the treatment of non small cell bronchogenic carcinoma. PMID- 16312836 TI - Surgery for cure of small cell bronchial carcinoma (SCLC) at early stages and postoperative combined modality treatment. PMID- 16312837 TI - Treatment of gastrointestinal tumors with MAb17-1A. PMID- 16312838 TI - Treatment of advanced colorectal and gastric cancer with 5-fluorouracil and calcium n-methyltetrahydrofolate. PMID- 16312839 TI - Treatment of unresectable nonsmall-cell lung cancer (NSCLC). PMID- 16312840 TI - Perfusion chemotherapy for colorectal liver metastases: a randomized study comparing FUDR against 5-FU/BCNU. PMID- 16312841 TI - Clinical pharmacology of cisplatin after intraperitoneal administration in patients with malignant ascites. PMID- 16312842 TI - Intraperitoneal chemotherapy in ovarian carcinoma with the combination of carboplatin and 5-fluorouracil. PMID- 16312843 TI - Adjuvant immunotherapy for cervical cancer. Two randomized controlled studies using OK-432 or sizofiran. PMID- 16312844 TI - 4-year experience in conservative management of muscle-infiltrating bladder cancer. PMID- 16312845 TI - Endocrine manipulation of human prostatic cancer: state of the art. PMID- 16312846 TI - Use of the LHRH analogue zoladex depot alone and in combination with cyproterone acetate in the treatment of advanced prostatic cancer. PMID- 16312847 TI - Leuprolide (TAP-144)--a general review. PMID- 16312848 TI - Treatment of prostatic cancer with TAP-144-SR, a depot preparation of LH-RH agonist (leuprolide). PMID- 16312849 TI - LH-RH analogues: review of international clinical experience. PMID- 16312850 TI - Free plasma amino acids in patients with multiple myeloma. PMID- 16312851 TI - Melphalan kinetics in hyperthermic isolation perfusion of the extremities. PMID- 16312852 TI - Neoadjuvant chemotherapy for localized Ewing's sarcoma. Preliminary results of a new protocol which uses surgery alone or followed by radiotherapy for local control. PMID- 16312853 TI - Kinetics of sex hormones after administration of 7.5 or 3.75 mg depot leuprolide including noncompliant patients and escape kinetics after a single-dose treatment. PMID- 16312854 TI - Spindle-cell sarcoma of the female breast. PMID- 16312855 TI - Comparison of MOPP/MOPP +/- RT vs. MOPp/ABVD +/- RT in poor prognosis Hodgkin's disease. Long-term results. PMID- 16312856 TI - Cyclophosphamide, epirubicin, vincristine, and prednisone (CEOP) in the treatment of advanced stage non-Hodgkin's lymphomas with poor histology. PMID- 16312857 TI - Results of three polychemotherapy programs in non-Hodgkin's lymphomas. PMID- 16312858 TI - Salvage combination chemotherapy with cisplatinum, etoposide and bleomycin in refractory non-Hodgkin's lymphomas. PMID- 16312859 TI - Alpha interferon in the treatment of hairy cell leukemia. PMID- 16312860 TI - Recombinant alpha 2 interferon in the treatment of chronic myeloproliferative disorders. PMID- 16312861 TI - Mitoxantrone and cytosine arabinoside (ARA-C) in adult acute leukemia: an interim report. PMID- 16312862 TI - VAD for remission induction as a first line and in relapsed adult acute lymphoblastic leukemia (ALL). PMID- 16312863 TI - Statistical evaluation of prognostic factors in childhood acute lymphoblastic leukemia. PMID- 16312864 TI - Low dose cytosine arabinoside in the treatment of acute non-lymphocytic leukemias and myelodysplastic syndromes. PMID- 16312865 TI - Treatment of polycythemia vera with hydroxyurea. PMID- 16312866 TI - Antiemetic therapy in patients treated with cisplatin chemotherapy. PMID- 16312867 TI - Early impairment of kidney function after cisplatin administration. PMID- 16312868 TI - Predicting toxicity during 5-fluorocytosine (5-FC) therapy by plasma level monitoring: a lesson to learn from cytostatic 5-fluorouracil (5 FU) pro-drugs. PMID- 16312869 TI - A 99mTc-DTPA study on platinum chronic nephrotoxicity: also glomerular lesions? PMID- 16312870 TI - Septicemias in cancer patients: incidence and outcome in relation to the underlying disease and infectious agent. PMID- 16312871 TI - Bactericidal activity of subinhibitory concentrations of mezlocillin and azlocillin in blood of patients with chronic lymphatic leukaemia. PMID- 16312872 TI - Imipenem/cilastatin as initial single-agent therapy for infections in cancer patients with neutropenia. PMID- 16312873 TI - Prophylactic administration of oral norfloxacin for prevention of gram-negative bacterial infections in patients with acute leukemias. PMID- 16312874 TI - Characteristics of coagulase-negative staphylococci isolated from septic leukemic patients. PMID- 16312875 TI - Antimycotic combination therapy in patients with acute myeloblastic leukemia. PMID- 16312876 TI - Thoracic herpes zoster treated with intravenous Acyclovir in three cancer patients. PMID- 16312877 TI - Preventive antibiotic therapy in surgical oncologic practice: cefotetan versus cefotaxime + imidazole. PMID- 16312878 TI - Immunoprophylaxis with thymostimulin in oncological surgery. PMID- 16312879 TI - Immunity during monochemotherapy with mitoxantrone: preliminary results. PMID- 16312880 TI - Pimafucin (natamycin) oral drops in the treatment of fungal infections of the oral cavity in children with chronic blood diseases. PMID- 16312881 TI - Human immunoglobulins in the prophylaxis of infections in granulocytopenic neoplastic patients. PMID- 16312882 TI - [Talking about acupuncture characteristics]. PMID- 16312883 TI - [Optimum selection of acupuncture treatment programs of cerebral infarction and its effects on insulin resistance]. AB - OBJECTIVE: To study the optimum acupuncture treatment program and the mechanism for treatment of cerebral infarction. METHODS: Sixty-three cases were randomly divided into 9 groups. The four factors, times of manipulation, the retaining time of the needle, acupuncture instrument and acupoints,and their corresponding three levels were adopted respectively in treatment of each group. Then nerve function defect score and insulin resistance were observed before and after treatment. RESULTS: The acupoints, the times of manipulation and the retaining time of the needle have significant effects on nerve function defect and insulin resistance (P < 0.01 or P < 0.05), and the acupuncture instrument has a significant effect on insulin resistance (P < 0.01). The choice of acupoints was the most important factor for acupuncture treatment of cerebral infarction. CONCLUSION: Acupuncture for regulating The Governor Vessel with twice manipulations and retaining the needle for 60 min is optimum treatment program for cerebral infarction. The good regulating effect of acupuncture on insulin resistance is one of the mechanisms of achieving the therapeutic results. PMID- 16312884 TI - [Acupuncture treatment for persistent vegetative state after operation of acute subdural hematoma]. AB - OBJECTIVE: To explore the therapeutic effect of acupuncture on the basis of western medicine treatment on persistent vegetative state. METHODS: The patients of treatment group were treated by acupuncture at "Liming", Shuigou (GV 26), Yintang (EX-HN 3), Shixuan (EX-UE 11), Yamen (GV 15), Yongquan (KI 1) on the basis of routine treatment of western medicine, more using the method for activating brain and restoring consciousness, and firstly using moxibustion until the manifestation of deficiency improving and then using above methods for prostration syndrome. And control group were treated with routine treatment of western medicine. RESULTS: The recovery rate and cured rate for mind after 3 therapeutic courses were 90.0% and 5.0%, and after 6 courses were 95.0% and 25.0% in the treatment group, respectively, which were significantly better than 47.5%, 0, and 65.0%, 5.0% in the control group. CONCLUSION: Acupuncture can obviously increase the recovery rate and cured rate for mind of persistent vegetative state. PMID- 16312885 TI - [Treatment of 58 cases of vitiligo by electro-plum-blossom needle therapy combined with catgut implantation at acupoints under TDP radiation]. AB - OBJECTIVE: To search for a method for increasing therapeutic effect on vitiligo. METHODS: One hundred and sixteen cases were randomly divided into a treatment group and a control group, 58 cases in each group. The treatment group were treated with electro-plum-blossom needle therapy plus catgut implantation at local acupoints under TDP radiation, and the control group with external application of sicorten. Their short-term and long-term therapeutic effects were observed. RESULTS: The short-term total effective rate was 98.3% in the treatment group and 74.1% in the control group with a significant difference between the two groups (P < 0.05). CONCLUSION: Electro-plum-blossom needle therapy plus catgut implantation at acupoints has a better therapeutic effect on vitiligo with no adverse effect. PMID- 16312886 TI - [Observation on therapeutic effect of acupuncture and moxibustion combined with sacral injection on prolapse of lumbar intervertebral disc]. AB - OBJECTIVE: To search for an effective method for treatment of prolapse of lumbar intervertebral disc. METHODS: Ninety-four cases were randomly divided into a treatment group of 48 cases and a control group of 46 cases. The treatment group were treated by acupuncture and moxibustion combined with sacral injection and the control group with simple sacral injection. Their therapeutic effects were compared. RESULTS: The cured and total effective rates were 64.6% and 97.9% in the treatment group, and 37.0% and 89.1% in the control group, with statistically significant differences between the two groups (both P < 0.05). CONCLUSION: Acupuncture and moxibustion combined with sacral injection has a significant therapeutic effect on prolapse of lumbar intervertebral disc. PMID- 16312887 TI - [Clinical observation on acupuncture combined with medication for intractable angina pectoris]. AB - OBJECTIVE: To compare clinical effects of acupuncture combined with medication and simple medication on intractable angina pectoris (IAP). METHODS: Seventy cases of IAP were randomly divided into two groups, a control group and a treatment group, 35 cases in each group. The control group were treated with nitric acid ester, beta-receptor blocking agent, calcium antagonist, aspirin and hypo-molecular heparin, and the treatment group were treated with acupuncture at Danzhong (CV 17) and Neiguan (PC 6) and others, once daily for 10 consecutive days on the basis of the above treatment. RESULTS: The total effective rates for symptoms and electrocardiogram were 88.6% and 62.9% in the treatment group and 60.0% and 31.4% in the control group, respectively, with a significant difference between the two groups; the incidence of acute heart attack and sudden death of 5.7% in the treatment group was lower than 20.0% in the control group, with no serious adverse effect. CONCLUSION: Acupuncture combined with medication is safe and effective for intractable angina pectoris and it can improve short-term prognosis. PMID- 16312888 TI - [Comparison and analysis on therapeutic effects of acupuncture plus massage therapy and drug on infantile diarrhea]. AB - OBJECTIVE: To compare therapeutic effects of acupuncture plus massage therapy and western medicine on infantile diarrhea. METHODS: A total of 120 cases of infantile diarrhea were randomly divided into a treatment group of 80 cases and a control group of 40 cases. The treatment group were treated by acupuncture and massage therapy, and the control group by smecta. RESULTS: The cured rate of 55.0% in the treatment group was better than 35.0% in the control group (P < 0.05). CONCLUSION: Acupuncture plus massage therapy has obvious therapeutic effect on infantile diarrhea. PMID- 16312889 TI - [Clinical study on acupuncture for treatment of 80 cases of simple obesity]. AB - OBJECTIVE: To search for an effective method for treatment of obesity and to study the mechanism of acupuncture slimming treatment. METHODS: The cases of obesity were randomly divided into a diet group of 40 cases and an acupuncture group of 80 cases. Their therapeutic effects and changes of TC, TG, LDL-C, HDL-C contents before and after treatment were observed. RESULTS: The effective rate of 76.3% in the acupuncture group was better than 27.5% in the diet group, and acupuncture could decrease serum TC, TG, LDL-C contents and increase HDL-C content. CONCLUSION: Acupuncture combined with diet therapy is an effective method for reducing weight at present, and metabolism of lipids is involved in acupuncture slimming mechanism. PMID- 16312890 TI - [Clinical study on treatment of fibromyalgia syndrome with penetration needling at the back]. AB - OBJECTIVE: To compare therapeutic effects of penetration needling at the back and western medicine on fibromyalgia syndrome (FS). METHODS: Thirty-eight cases were randomly divided into the observation group treated with penetration needling at the first and second lines of The Bladder Meridian of Foot-Taiyang and Governor Vessel, and the control group treated mainly with oral administration of amitriptyline, 19 cases in each group. RESULTS: There were significant differences in the cured-markedly effective rate and the results of follow-up for 6 months for the cured-markedly effective cases between the observation group and the control group (all P < 0.01). CONCLUSION: Penetration needling therapy in the therapeutic effect on fibromyalgia syndrome is better than the medical treatment. PMID- 16312891 TI - [Observation on therapeutic effect of acupuncture and moxibustion in 60 cases of Sjogren syndrome]. AB - OBJECTIVE: To observe clinical therapeutic effect of acupuncture on Sjogren syndrome. METHODS: Treatment group (n=60) were treated with acupuncture of clearing away dryness and toxic substance and removing obstruction in collaterals at Quze (PC 3), Taichong (LR 3), Xuehai (SP 10), Sanyinjiao (SP 6) and Taixi (KI 3), and the control group (n=60) with prednisone. Their therapeutic effects were compared. RESULTS: The total effective rate was 73.3% in the treatment group and 56.7% in the control group with a significant difference between the two groups (P < 0.05); there was no reverse effect in the treatment group. CONCLUSION: The needling method for clearing dryness and removing obstruction in collaterals is effective and safe for treatment of Sjogren syndrome. PMID- 16312892 TI - [Acupunture and moxibustion evidence-based treatment method for periarthritis of shoulder]. AB - OBJECTIVE: To introduce to establishment method of evidence-based acupuncture and moxibustion treatment program for periarthritis of shoulder from the point of view of evidence-based medicine. METHODS: One patient of periarthritis of shoulder was used as an example, and it was introduced how to raise clinical questions, to retrieval relative literature, to evaluate the retrieval literature, and at last, how to use the retrieval evidences to stipulate treatment program in combination with concrete condition of this case and the doctor's experience. RESULTS: The acupuncture and moxibustion treatment program for periarthritis of shoulder established by evidence-based medicine was adopted to treat the patient for 15 sessions, as a result the life quality of the patient improved obviously. CONCLUSION: Through clinically raising questions, searching for evidences, analyzing evidences, making a strategic decision, practising evidence-based course, the patient can attain satisfactory therapeutic effect, and the physician can raise theoretical level and clinical ability. PMID- 16312893 TI - [Clinical observation on acupuncture for treatment of depression]. AB - OBJECTIVE: To search for the best therapy for depression. METHODS: The acupuncture group of 34 cases were treated by acupuncture at acupoints on the head as main and acupoints of the limbs as adjuvant, two treatments a week, and the control group of 30 cases by antidepressant deanxit for 5 weeks. RESULTS: There was no significant difference between the two groups in the therapeutic effect (P > 0.05). CONCLUSION: Acupuncture and western medicine deanxit have a similar therapeutic effect on depression. PMID- 16312894 TI - [Experimental study on conduction of Gong tonality vibromusic sound wave in the healthy human body]. AB - OBJECTIVE: To study the conduction of Gong tonality vibromusic sound wave along meridians in healthy human body, and investigate differences of the sensitivity of different meridians and genders to this vibromusic message. METHODS: Emit the Gong tonality music signal under the water and then investigate the responses of different acupoints and control points at the tissue of the same level to the vibromusic sound wave. RESULTS: There were differences of sensitivity to music waves at source acupoints on the foot, sensitivity of Zusanli (ST 36) was significantly higher than its control point (P < 0.05), and there was difference between genders in the sensitivity of Sanyinjiao (SP 6) and Yinlingquan (SP 9) to music sound wave. CONCLUSION: Gong tonality vibromusic sound wave can conduct along meridians in healthy human body, and there are differences between different meridians and different genders in the sensitivity to the music sound wave. PMID- 16312895 TI - [Relationship of meridians with human material and energy systems study on theory and clinical application of meridians (IV)]. AB - OBJECTIVE: To expound the relationship between meridians and the human two information systems, material system and energy system. METHODS: Summarize and analyze the results of experimental and other studies. CONCLUSION: The carrier of the meridian is human material system. The nerve, body fluid, microcirculation and chemical factors take part in meridian circulation, and it must depend on the energy information system which works mainly at the form of electromagnetic and chemical oscillation with functions of physiological regulation and pathological repair. Oscillation parameters include various signals of regulating development and physiologic activities, and coordinate functions of organs and tissues. The meridians have the "quantum" characteristics, material particles coexist and exchange each other with electromagnetic and chemical oscillation characteristic. PMID- 16312896 TI - [Experimental study on influence of acupuncture and moxibustion therapy on apoptosis of colonic epithelial cells in rats of ulcerative colitis]. AB - OBJECTIVE: To study the mechanism of acupuncture and moxibustion in adjusting apoptosis of colonic epithelial cells of the rat of ulcerative colitis (UC). METHODS: UC rats were randomly divided into four groups including normal control group (NC), model control group (MC), herbs-partitioned moxibustion group (HPM) and electroacupuncture group (EA). The HPM and EA groups were respectively treated with herbs-partitioned moxibustion and electroacupuncture at "Qihai" (CV 6) and bilateral "Tianshu" (ST 25). Then the colon of the rat was taken and the changes of the tissue structure and the cell apoptosis were observed by electron microscope and flow cytometer respectively. RESULTS: The colon tissue showed histopathological changes, and apoptosis of epithelial cells increased in the model mice of UC as compared with the normal rat. Electroacupuncture and drug separated moxibustion could significantly decrease the abnormal apoptosis of epithelial cells. CONCLUSION: The adjustment of the abnormal apoptosis of epithelial cells is one of the mechanisms of drug-separated moxibustion and electroacupuncture treating UC. PMID- 16312897 TI - [Change rate in bone cells of mice]. AB - OBJECTIVE: To study the anti-mutation action of acupuncture and moxibustion. METHODS: Mice were randomly divided into 6 groups, group 1 (normal control group), group 2 (positive control group), group 3 (prevention group I), group 4 (prevention group II ), group 5 (treatment group I) and group 6 (treatment group II). The mice in the group 2-6 were treated by cyclophosphamide (ip, 50 mg/kg body weight), and in the 3'-6 groups were given acupuncture at "Zusanli" (ST 36) and moxibustion at "Guanyuan" (CV 4). At the end of experiment, all the mice were decapitated and chromosome aberration rate and sister chromatid exchange rate of bone marrow cells were investigated. RESULTS: The chromosome aberration rate and the sister chromatid exchange rate of bone marrow cells in the positive control group increased significantly as compared with the normal control group, while they decreased significantly in the group 3, 4, 5, 6 as compared with the positive control group (P < 0.01). CONCLUSION: Acupuncture and moxibustion have anti-mutation action, inhibiting the increase of chromosome aberrations and sister chromatid exchange of bone marrow cells in mice induced by cyclophosphamide. PMID- 16312898 TI - [Clinical observation on acupuncture at Neimadian for analgesia postoperation of extremities]. AB - OBJECTIVE: To observe therapeutic effect of acupuncture at Neimadian on pain after operation of four limbs. METHODS: Sixty-two patients were randomly divided into an observation group and a control group, 31 cases in each group. The observation group were treated with electroacupuncture at Neimadian for 30 min, and the control group with oral administration of tramadoli hydrochloridum. Changes of pain within 24 hours were observed. RESULTS: The analgesic effect in the observation group was better than that in the control group (P < 0.05). CONCLUSION: Analgesic effect of acupuncture at Neimadian on pain after operation of four limbs is superior to that of oral administration of tramadoli. PMID- 16312899 TI - [Application of transcutaneous electric nerve stimulation in operation of thyroid tumors]. AB - OBJECTIVE: To study the effect of transcutaneous electric nerve stimulation for decreasing stress response and strengthening the anesthetic effect in the operation of thyroid tumors. METHODS: Sixty patients with thyroid tumors were randomly divided into an observation group and a control group, 30 cases in each group. All of them were given modified one-point cervical plexus nerve obstruction. The stimulation at bilateral Hegu (LI 4) and Quchi (LI 11) with HANS was added to the observation group from the beginning to end of operation. RESULTS: The blood pressure, heart rate and jerking reflection were decreased significantly as compared with the control group. The oxygen saturation and respiratory frequency did not change. CONCLUSION: Cardiovascular stress response can be decreased with HANS in the operation of thyroid tumors and anesthetic effect can be strengthened. PMID- 16312900 TI - [Study on the theory of acupoint functions]. AB - OBJECTIVE: To explore the basic concept of functions of acupoints. METHODS: The principles of Chinese medicine therapy and acupuncture therapy are analyzed and compared and it is pointed out that there is an essential difference between the functions of traditional Chinese medicine and acupoints. Chinese medicine correct the deviation using their deviation property, but acupoints correct the deviation using their bi-directional regulative actions. RESULTS: Acupuncture and moxibustion therapy is different from Chinese medicine in treating syndromes of cold, heat, deficiency and excess. That the descriptive way of traditional Chinese medicine functions is applied to those of acupoints is somewhat one sideness. CONCLUSION: Acupoints have their own characteristics and principles of application, and acupoints have the effects of close therapy, distal therapy and special effects, and action law of special acupoints. Therefore, induction for functions of acupoints should supplement and perfect these functions. PMID- 16312901 TI - [Analysis on characteristics of ancient acupuncture and moxibustion treatment of tuberculosis consumptive diseases]. AB - Systematical and comprehensive statistics, induction and analysis on literature and data of acupuncture and moxibustion treatment of tuberculosis in incunabula are carried out by computer. The results indicate that in treatment of tuberculosis, most select acupoints of the upper back, chest-epigastrium and The Yangming Channel on the leg to promote dispersing function of the lung, strength the spleen and anti-tuberculosis; select acupoints of The Conception Vessel on the lower abdomen and Shenshu (BL 23) and others on the lower back to tonify the kidney and anti-tuberculosis. Clinically, most select moxibustion and acupuncture with reinforcing-reducing manipulation and pricking blood therapy. And referential recipes for clinical treatment are raised. PMID- 16312902 TI - [Theoretical basis of regulating mind and relieving the depressed liver needling method for treatment of depression]. AB - The mechanism of depression attack was studied from the point of view of TCM theory, literature and clinical practice. It is held that the brain and liver are the key zang- and fu-organs for attack of depression, and it is first put forward that dysfunction of mind and the liver failing in soothing and regulating the circulation of qi are the TCM pathogenesis of this disease. Needling method for regulating mind and soothing the liver is established with the acupoints of The Governor Vessel and The Liver Channel selected mainly. PMID- 16312903 TI - [Advances of clinical studies of acupuncture and moxibustion for treatment of rheumatoid arthritis]. AB - OBJECTIVE: To summarize recent development of studies on acupuncture and moxibustion for prevention and treatment of rheumatoid arthritis, so as to provide a basis and thinking for clinical scientific studies. METHODS: More than 60 papers about clinical and experimental studies were reviewed, choosing clinically commonly-used acupoints and introducing mainly different therapeutic methods. RESULTS: Acupuncture and moxibustion can prevent and cure rheumatoid arthritis with outstanding results. CONCLUSION: Acupuncture and moxibustion have vast vistas for treatment of rheumatoid arthritis. PMID- 16312904 TI - [Factors influencing antibiotics prescribing of primary health physicians in acute upper respiratory infections]. AB - OBJECTIVES: To explore the factors influencing antibiotics prescription by primary health physicians for acute upper respiratory infections (URI). METHODS: We performed a survey of 370 primary health physicians randomly sampled in April, 2003. The questionnaire consisted of a prescription on the scenario of acute bronchitis case, along with opinions and reasons for prescribing antibiotics on URI. RESULTS: We found that 54.7% of the physicians prescribed antibiotics on the example case of acute bronchitis which is known as not needing antibiotics. Female physicians and ENT physicians had a greater tendency to prescribe antibiotics. The factors influencing antibiotics prescription on URI were the belief about the effectiveness of antibiotics, preference for their own experiences rather than clinical guidelines, perception of patients' expectations, and perception of competitive environment. The prescription of antibiotics in the example case was affected by how much they usually prescribe antibiotics (OR = 2.400, 95% CI = 1.470-3.917) and the physicians who thought that antibiotics were helpful for their income prescribed antibiotics more than others (OR = 6.773, 95% CI = 1.816-25.254). CONCLUSION: These findings demonstrated that the false belief on the effectiveness of antibiotics, patient's expectation of medication and fast relief of symptoms, and perception of competitive environment all affected the physicians' prescription of antibiotics on URI. It may help to find barriers to accommodate scientific evidence and clinical guidelines among physicians and to specify subgroups for education about appropriate prescription behaviors. PMID- 16312905 TI - [Medical expenses by site of cancer and survival time among cancer patients in the last one year of life]. AB - OBJECTIVES: To analyze medical expenses by cancer site and survival time among cancer patients in their last year of life. METHOD: The study subjects were 45,394 people that had died of cancers in 2002, were registered by the Korea Central Cancer Registry and received National Health Insurance benefit in the last year (360 days) of life. Personal identification data, general characteristics, dates of death and cancer incidence, and site of cancer were collected from the National Statistical Office and the Korea Central Cancer Registry, and merged with the data of the individual medical expenses of the Health Insurance Review Agency. RESULTS: Average monthly cost curves were U shaped with high costs near the time of diagnosis and death, and lower costs in between. Medical expenses in the last year of life were around 30.3, 16.7, 13.0, and 12.1 million won among leukemia, lymphoma, ovarian cancer, and breast cancer patients, respectively. Digestive organ cancers including stomach, esophagus, liver, pancreas, and colorectal cancers had relatively low medical expenses. Medical expenses in the last year of life were inverse Ushaped with high expenses near one year of survival. Average monthly cost in the 12 months before death among the patients who had survived 10-15 years were more than two-fold greater than the cost before diagnosis among those who had survived for less than one year. CONCLUSIONS: Leukemia was the most expensive cancer. It is possible that once diagnosed as cancer, medical expenses do not return to the level before diagnosis. Further research will be needed to understand the magnitude and change of the medical expenses among cancer patients with long term follow up data. PMID- 16312906 TI - [Determining factors of intention to actual use of charged long-term care services for the aged]. AB - OBJECTIVES: To help develop strategies to cope with the changes arising from the rapid aging process by predicting the determining factors of intention to actual use of the charged long-term care services for elderly as perceived by the middle aged who play the major role of supports. METHODS: Subjects were the parents (men 177, women 507) in their 40s of the students selected from a university of Busan city. A questionnaire survey was conducted for 4 weeks in October 2003 about the knowledge for long-term care service, the intention of actual use, and the preferences about the type of service suppliers. Data analysis was performed with frequency, chi-square test, and t-test using SPSS program (ver 10.0K), along with data mining using decision tree of Enterprise Miner V8.2 by SAS. RESULTS: About half of the subjects (53.7%) had the actual experiences of elderly supports. Intentions to use the charged services were relatively high in home visiting nursing care service (40.1%) and long-term care facilities service (40.4%), and were influenced by previous knowledge about the services. The intentions were stronger in women, those with higher education, and those with greater income levels. Actual elderly supports were mc (80%) done by women, and the perceived burdens for supports were bigger in women and those of lower s economic level. Desired charges were about 10,000 for the bath service, 20,000 won for the rests services day, and about 500,000 won for the long-term care facil service per month. From the result of decision analysis, the job professionalism was the most impol determining factor of intention to actual use of the serv with validation as 63-71%. Health and welfare mixed facilities were preferred, and the most impor consideration was the level of professionalism. CONCLUSIONS: Intention to actual use of the chad services was largely determined by the aspects of time cost. Polices to increase the number of service supp and to decrease the burdens perceived by ac supporters were strongly recommended. PMID- 16312907 TI - [Epidemiology of psychosocial distress in Korean employees]. AB - OBJECTIVE: To estimate the magnitude of psychosocial distress and examine eligible factors associated with the development of psychosocial distress in Korean employees, using a nationwide sample. METHODS: A total of 6977 workers were recruited from 245 companies. A structured questionnaire was used to assess sociodemographics, health-related behaviors, job characteristics, social support at work, personality traits (locus of control, type A behavior pattern), self esteem, and psychosocial distress. RESULTS: The results showed that 23% of workers were categorized as high stress, 73% as moderate, and 5% as normal. Hierarchical multiple regression analysis showed that psychosocial distress was more common in younger workers, both male and female. Regular exercise was negatively associated with increase of psychosocial distress. In job characteristics, as expected, low decision latitude, high job insecurity, and low social support at work were related to high psychosocial distress. Personality traits such as locus of control and type A behavior pattern, and self-esteem were more powerful predictors of psychosocial distress than general characteristics, health-related behavior, and job characteristics. There were some gender differences. While men who are less educated and single (unmarried, divorced, and separated) experienced higher levels of psychosocial distress than those who are educated and married, women who feel high job demand experienced higher levels of psychosocial distress than those who feel low job demand. CONCLUSIONS: The proportion of the high stress group was higher than expected, and psychosocial factors like social support and personality characteristics (e.g. locus of control, type A behavior pattern and self-esteem) were more significant factors for psychosocial distress than other variables. This finding suggests that some psychosocial factors, especially inadequate social support, low self-esteem and lack of internal locus of control for the development of psychosocial distress, will also operate as an intervention strategy in the worksite stress reduction program. It is strongly required that worksite stress reduction programs should be established in at both occupational and level as well as in individual levels. PMID- 16312908 TI - [Mortality among medical doctors based on the registered cause of death in Korea 1992-2002]. AB - OBJECTIVE: To compare the mortality rate of Korean medical doctors to that of the general Korean population for the period 1992-2002. METHODS: The membership records of the Korean Medical Association were linked to the 1992-2002 death certificate data of Korea's National Statistical Office using 13-digit unique personal identification numbers. The study population consisted of 61,164 medical doctors with a follow-up period of 473,932 person-years. Standardized mortality ratios(SMRs) were calculated to compare cause-specific mortality rates of medical doctors to those of the general population. RESULTS: We confirmed 1150 deaths at ages from 30 to 75 years from 1 January 1992 to 31 December 2002. The SMR for all cause of death was 0.47(95% CI: 0.44-0.50). The SMRs for smoking-related diseases such as cerebrovascular accidents and chronic obstructive pulmonary disease were smaller than the SMR of all-cause of death. However, the SMRs for colorectal and pancreatic cancers were not significantly lower than those of the general population. Transport accidents and suicides accounted for 72% (94 of 131) of external causes of death. The SMR for suicide was 0.51 (95% CI: 0.38-0.68). CONCLUSIONS: The mortality rate of South Korean medical doctors was less than 50% that of the general population of South Korea. Cause-specific analysis showed that mortality rates in leading causes of death were lower among medical doctors although differences in mortality rates between medical doctors and the general population varied with the causes of death. These health benefits found among medical doctors may be attributable to the lower level of health damaging behaviors (e.g., lower smoking rates) and better working conditions. PMID- 16312909 TI - [Fatigue subjective symptoms and risk factors in bank workers with VDT]. AB - OBJECTIVES: [corrected] To investigate the fatigue subjective symptoms and VDT related risk factors in bank workers. METHODS: A total of 2178 workers (62.8% male, 37.2% female) were surveyed with a self-administered and structured questionnaire consisting of 30 fatigue symptoms and other VDT related factors regarding work characteristics, work postures, work environments, personal medical histories, etc. Fatigue subjective symptoms were divided into 3 groups of 10 questions each: a dull, drowsy and exhausted feeling (Group I), a mental decline of working motivation (Group II), or a feeling of incongruity in the body and dysfunction of autonomic nervous system (Group III). Each question was weighted as 0 for 'none', 1 for 'sometimes', and 2 for 'always'. RESULTS: Mean age was 35.8 +/- 7.2 years (38.9 +/- 6.0 in males, 30.6 +/- 5.8 in females), Mean work duration was 7.7 +/- 7.1 years (8.4+7.4 in males, 6.5 +/- 6.4 in females). Mean symptom score in males was 11.0 +/- 8.1 (4.9 +/- 3.1 in Group I, 3.0 +/- 3.1 in Group II, 3.1 +/- 2.8 in Group IIl) and in females was 17.5 +/- 9.5 (7.0 +/- 3.8 in Group I, 4.6 +/- 3.5 in Group II, 5.9 +/- 3.4 in Group III). Mean scores were higher in the order of Group I > III > II. Females had significanntly higher scores than males in all three groups (p < 0.001). Most common symptoms complained of as 'always' or sometimes' were 'feel strained in the eyes' (85.4%) among Group I, 'find difficulty in thinking' (54.2%) amongGroup II, and 'feel stiff in the shoulders' (72.3%) among Group III. In multiple regression analysis, female, non-straight spine at workstation, more VDT work hours, history off physical therapy, glare of screen, overtime work, youngage, and non-horizontal elbow position were significantly related with high score of symptoms. CONCLUSIONS: More supportive VDT work environment and education for correct work postures for VDT workers are recommended. PMID- 16312910 TI - [Association of Internet addiction with health promotion lifestyle profile and perceived health status in adolescents]. AB - OBJECTIVES: To identify the relationship between the internet addiction of adolescents and their Health Promotion Lifestyle Profile and Perceived Health Status, and thereby to detect the impact of internet addiction on the health of adolescents, produce the basic information necessary to develop a prevention program for internet addiction and to plan for a health promotion program. METHODS: This study was designed as a cross-sectional study, and the subjects were the second-grade students of three junior-high and three high schools located in the city of K in Kyung Gi Province. Out of 769 subjects, 764 completed the questionnaires (99.3%); 369 (48.3%) junior-high school students and 395 (51.7%) high school students. The questionnaires were composed of Young's Internet Addiction, Health Promotion Lifestyle Profile, Perceived Health Status, and general characteristics. We used t-test, ANOVA in means comparison between groups, chi2-test in frequency analysis, and multiple regression analysis in multivariate analysis, using the SAS 8.1 program. RESULTS: There was a statistically significant difference in Health Promotion Lifestyle Profile according to internet addiction status (severe addiction vs. other status, P < 0.0001). The Perceived Health Status scores was lowest in the severe addiction group (P < 0.001). There was also a significant negative correlation between internet addiction and Health Promotion Lifestyle Profile (P < 0.0001). The results of multiple regression showed that Young's Addiction Score was significant for the subjects' Health Promotion Lifestyle Profile after controlling for other variables (P < 0.0001). CONCLUSIONS: This study showed that the severe internet addiction group had the lowest score in Health Promotion Lifestyle Profile and Perceived Health Status, which suggests that the addiction could have a negative effect on the health status of adolescents. PMID- 16312911 TI - [Development of measurement scale for the quality of life in hypertensive patients]. AB - OBJECTIVES: To develop a tool for multidimensional measurement of the quality of life, which was psychometrically sound, short, and easy to administer for patients with hypertension. METHODS: A sample of 1115 hypertensive patients aged 20 or above in Cheong-Song County was studied from June 1997 to October 1998. In the development of the instrumental stage, the authors first conceptualized the quality of life. Item generation, item reduction, and questionnaire formatting were followed. Item-level (item descriptive, missing%, item internal consistency, item discriminant validity) analysis, scale-level (scale descriptive, floor and ceiling effect) analysis, and other tests(Cronbach's alpha, inter-dimension correlations, factor analysis, clinical validity) were performed to evaluate the validity and reliability of the new measurement scale. After 1 year, responsiveness and confirmatory factor analysis were performed. RESULTS: The results of both item-level and scale-level analyses were acceptable. An acceptable degree of internal consistency was observed for each of the dimensions (Cronbach's alpha was 0.60 or higher). Inter-dimension correlations were below 0.50 and the factor analysis result was the same as the intended dimension structure. Correlation coefficients between perceived health status, stress and dimensions were proven to be acceptable. The result of comparing dimensional score means among ADL and MMSE-K groups above 60 years was statisticalIly significant (p < 0.05). The result of confirmatory factor analysis concluded that the dimensional structure model was well fitted. However, the result of responsiveness test using sensitivity and specificity was unsatisfactory. CONCLUSIONS: The newly developed measurement scale is psychometrically reliable and valid instrument for measuring quality of life in hypertensive patients. PMID- 16312912 TI - [Correlates of self-rated fatigue in Korean employees]. AB - OBJECTIVE: To elucidate the correlates of self-rated fatigue in Korean employees. METHODS: The data for 10,176 (men, 7984; women, 2192; mean age, 34.2; SD: 8.8) employees recruited from a nation-wide sample were examined. A structured questionnaire was used to measure the participants' fatigue, sociodemographics (sex, age, education, and marital status), job-related characteristics (work duration, grade at work, work hours, shiftwork, employment type, and magnitude of workplace), and health-related habits (smoking, drinking, coffee intake, and exercise). Two types of measurement for fatigue were used to evaluate the magnitude of fatigue: self-rated question and a standardized measurement tool (Multidimensional Fatigue Scale: MFS). RESULTS: According to the self-rated fatigue, 32% of employees reported that they felt fatigue for the past two weeks, and 9.6% of males and 8.7% of females had experienced excessive fatigue (6 months or more). Hierarchical multiple regression analysis showed that fatigue measured by MFS was more common in women, younger, college or more graduated, single, and employees who do not regularly exercise. Fatigue was also associated with long work hours, and the size of the workplace (< 1000 employees). CONCLUSIONS: These results suggest that fatigue has been considered as a common complaint, and that it is affected by job-related factors like work hours and the workplace size as well as sociodemographics or health-related behaviors. Further research is needed to clarify the effects of fatigue on adverse health,outcomes, work performance, work disability, sick absence and medical utilization, and to examine the relationship of job characteristics (e.g.: work demand, decision latitude) to fatigue. PMID- 16312913 TI - [Analysis of behavioral stage in pap testing by using transtheoretical model]. AB - OBJECTIVE: To evaluate the relationships among sociodemographic characteristics, health behaviors, levels of pros and cons and stages of change in Pap testing for uterine cervical cancer. METHODS: A questionnaire survey was performed on 560 randomly sampled people who were assigned to participate in a Pap testing program by the 'National Cancer Screening Project in 2003' between 25 September and 10 October in Gyeonggi, Korea. Data about the behaviors and intentions of Pap testing, sociodemographic characteristics, health behaviors, and levels of acknowledged benefit (pros) and barrier (cons) for Pap testing was collected. The stages of change were grouped according to behaviors and intentions of Pap testing as passive, active, and relapse. RESULTS: Logistic analysis between the passive and active groups showed that city dwellers, 'high' and 'middle' groups in terms of the individual's health belief, those who had undergone a health examination within the past 2 years, and those who had undergone hormone replacement therapy had a higher odds ratios to be in the active group. As the 'benefit' scores increased and the 'Unnecessity' scores decreased, the probabilities to be in the active group increased. According to the logistic analysis results between the active and relapse groups, those who were 60 years or older, members of the National Heath Insurance, and those who had not undergone a health examination within the past 2 years had a higher odds ratio to be in the relapse group. The 'Benefit' scores were not significant in this relationship. The probabilities of being in the relapse group increased as the 'Unnecessity' and 'Shamefulness' scores increased. CONCLUSIONS: In conclusion, health planners should inform women in the passive group of the benefits and necessity of Pap testing. It would be better to reduce the barriers to the active group of undergoing Pap smear. This study might be a useful guide for future planning of Pap testing program. PMID- 16312914 TI - [Participation rate and related socio-demographic factors in the national cancer screening program]. AB - BACKGROUND: Cancer is the leading cause of death and one of the largest burdens of disease in Korea. In 1996, the 'Ten year Plan for Cancer Control' was formulated and the government then adopted the plan as a national policy. As part of this plan, the National Cancer Screening Program (NCSP) for Medicaid recipients was formulated, and the government adapted this in 1999. For low income beneficiaries of the National Health Insurance Corporation (NHIC), the screening program has been in place since 2002. In 2002, the target cancers of NCSP were stomach, breast and cervical cancer. This study was conducted to examine the relationships between the participation rate, the abnormal screening rate and the socio-demographic factors associated with participation in the screening program. METHODS: To analyze the participation rate and abnormal rate for the NCSP, we used the 2002 NCSP records. The information on the socio demographic factors was available from the database of the beneficiaries in the NHIC and Medicaid. RESULTS: The participation rate of the Medicaid beneficiaries for the stomach, breast and cervical cancer screening were 9.2%, 15.5% and 15.0%, respectively, and 11.3% and 12.5%, except cervical cancer which wasn't be included in the NCSP, for the beneficiaries of the NHIC. The abnormal rate of stomach, breast and cervical cancer screening were 25.7%, 11.2% and 21.0%, respectively, for the beneficiaries of Medicaid and 42.6% and 19.4% for the beneficiaries of the NHIC. On the multiple logistic regression analysis, gender, age and place of residence were significantly associated with participation rates of the NCSP. For stomach cancer, women participated in the NCSP more than men. The participation rate was higher among people in their fifties and sixties than for those people in their forties and those people over seventy years in age. For the breast and cervical cancer, people in their fifties were more likely to participate in the NCSP than people in their forties and people over sixty. For the place of residence, people in the rural areas participated more than those people in any other places. CONCLUSIONS: The above results show that the participation rate and abnormal rate were significantly associated with the socio demographic factors. To improve the participation rate for the NCSP, more attention should be given to the underserved groups. PMID- 16312915 TI - [The association between serum GGT level within normal range and risk factors of cardiovascular diseases]. AB - OBJECTIVES: We conducted this study to examine the association between serum GGT levels within the normal range and the risk factors of cardiovascular diseases METHODS: We examined the cross-sectional association between serum GGT and the systolic and diastolic blood pressure, fasting blood glucose, total cholesterol, triglyceride, high density lipoprotein cholesterol (HDL-cholesterol), and uric acid among 975 subjects that participated in the health examination of a university hospital located in Daegu city. All the patients' GGT levels were within the normal range. RESULTS: After adjustment were made for age, body mass index (BMI), smoking status, drinking frequency, exercise frequency and coffee intake, the serum GGT level was positively associated with fasting blood glucose (p < 0.01), total cholesterol (p < 0.01), and triglyceride (p < 0.01) in men, and it was positively associated with fasting blood glucose (p < 0.01), total cholesterol (p < 0.05), triglyceride (p < 0.01), and uric acid (p < 0.01) in women. The associations were not significantly different depending on the status of alcohol drinking or obesity, except for the associations of serum GGT with diastolic blood pressure (P for interaction = 0.04) and uric acid (P for interaction = 0.04) between the lean and obese subjects. CONCLUSIONS: Serum GGT levels within the normal range were positively associated with fasting blood glucose, triglyceride and uric acid in most subgroups irrespective of the drinking or obesity status. These results suggest that GGT has important clinical implications as being more than just a marker of alcohol consumption and hepatobiliary disease. PMID- 16312916 TI - [Clinical study on needling method for regulating mental activities and soothing liver for treatment of melancholia]. AB - OBJECTIVE: To observe clinical therapeutic effect of needling method for regulating mental activities and soothing liver on melancholia. METHODS: One hundred and ten cases of depression were selected according to the diagnostic criteria of depression in mood disturbance (emotional and mental disturbance) and randomly divided into a regulating mental activities and soothing liver needling group (n=50), a traditional acupuncture group (n=30) and a prozac treatment group (n=30). Their therapeutic effects were evaluated by Hamilton depression scale, Self-rating depression scale (SDS), psychiatric self-rating scale (SCL-90). RESULTS: In the regulating mental activities and soothing liver needling group, 34 cases were cured, 10 cases were markedly effective, 4 cases were effective and 1 cases were ineffective, with the cured rate of 69.4% and the total effective rate of 98.0%, which were significantly better than those in the traditional acupuncture group (the cured rate was 48.3% and the total effective rate was 89.7%), and was not significantly different with the prozac treatment group (the cured rate was 64.0% and the total effective rate was 96.0%). CONCLUSION: The needling method for regulating mental activities and soothing liver has a better therapeutic effect on depression. PMID- 16312917 TI - [Effect of acupuncture combined with medicine on neuropeptide Y in the patient of hypertension]. AB - OBJECTIVE: To search for a method for increasing therapeutic effect on hypertension and study on the mechanism. METHODS: Seventy-five cases were randomly divided into the treatment group (n=45) treated by acupuncture plus medicine, and the control group (n=30) treated by medicine. Their blood pressure and plasma neuropeptide Y (NPY) before and after treatment were investigated. RESULTS: Blood pressure and NPY content in both the two groups decreased significantly (P < 0.01), and the treatment group in decreasing blood pressure and NPY content was superior to the control group (P < 0.05). CONCLUSION: Acupuncture and medicine have cooperation in treatment of hypertension, which is performed possibly through decreasing NPY. PMID- 16312918 TI - [Clinical observation on acupuncture for treatment of bronchial asthma at acute stage]. AB - OBJECTIVE: To observe effect of acupuncture on asthma at acute stage. METHODS: Ninety cases of asthma at acute stage were randomly divided into an observation group of 60 cases and a control group of 30 cases. Clinical therapeutic effect and the mechanisms were studied from symptoms and signs, pulmonary function and serum interleukin -4 (IL-4), gamma-interferon (INF-gamma). RESULTS: The pulmonary function was improved, the vital capacity was increased and attack of asthma was reduced in both the observation group and the control group, but the observation group was better than the control group in the pulmonary function improving degree (P < 0.05). CONCLUSION: Acupuncture can strengthen the relieving action of medicine on asthma and it is one of effective methods for bronchial asthma. PMID- 16312919 TI - [Observation on clinical therapeutic effect of acupuncture on apoplectic pseudobulbar palsy]. AB - OBJECTIVE: To find a clinically effective therapy for apoplectic pseudobulbar palsy. METHODS: Seventy-five cases of apoplectic pseudobulbar palsy were randomly divided into 3 groups, acupuncture group, western medicine group and acupuncture plus medicine group. They were treated for one month. RESULTS: The scores for clinical symptoms and signs, and the clinical scores for water provocative test in the acupuncture plus medicine group and the acupuncture group were superior to that in the western medicine group with significant difference (P < 0.01), and the therapeutic effects of the 3 groups were related with age, course of disease and condition of illness. CONCLUSION: Acupuncture has some predominances in treatment of apoplectic pseudobulbar palsy. PMID- 16312920 TI - [Clinical observation on 47 cases of intractable hiccup treated by acupuncture at Tianding (LI 17)]. AB - OBJECTIVE: To search for a convenient and effective needling method for treatment of hiccup. METHODS: Ninety-four cases were randomly divided into treatment group treated by acupuncture at Tianding (LI 17) and a control group treated by selected routine acupoints, 47 cases in each group. They were respectively treated based on differentiation of symptoms and signs for less than 10 sessions, and their therapeutic effects were compared. RESULTS: The therapeutic effects on primary excess syndrome of intractable hiccup in the two groups were superior to that secondary deficiency syndrome (both P < 0.05); there was no significant difference between the two groups in the therapeutic effect (P > 0.05). CONCLUSION: Acupuncture at Tianding (LI 17) is a convenient method for hiccup with a definite therapeutic effect. PMID- 16312921 TI - [Acupuncture combined with catgut embedding therapy for treatment of 158 cases of facial paralysis]. AB - OBJECTIVE: To observe therapeutic effect of acupuncture combined with catgut embedding therapy on peripheral facial paralysis. METHODS: Treatment group of 158 cases were treated by acupuncture combined with catgut embedding therapy and control group of 54 cases were treated by acupuncture, and their therapeutic effects were compared. RESULTS: There was no significant difference in the total effective rate (P > 0.05), but the cured rate in the treatment group was significantly higher than that in the control group (P < 0.01). CONCLUSION: Acupuncture combined with catgut embedding therapy is a better therapy for facial paralysis. PMID- 16312922 TI - [Clinical observation on yintang (Ex-HN 3) point-injection for treatment of obstinate hiccup]. AB - OBJECTIVE: To search for an effective method for treatment of obstinate hiccup. METHODS: Ninety cases of obstinate hiccup secondary to cerebral diseases were randomly divided into 3 groups, 30 cases in each group. Group A were treated by injection of compound chlorpromazine into Yintang (Ex-HN 3), group B by intramuscular injection of compound chlorpromazine and group C by acupuncture at Yintang (Ex-HN 3). Their therapeutic effects were compared. RESULTS: The effective rate was 93.3% in the group A, 10.0% in the group B and 30.0% in the group C, the group A being better than the group B and the group C (P < 0.01). CONCLUSION: Injection of compound chlorpromazine into Yintang (Ex-HN 3) has a significant therapeutic effect on obstinate hiccup. PMID- 16312923 TI - [Clinical observation on 136 cases of chilblains treated by acupuncture combined with massage]. AB - OBJECTIVE: To compare therapeutic effects of acupuncture plus massage and medicine on chilblains. METHODS: Two hundred and sixty-four cases of chilblains were randomly divided into an acupuncture group of 136 cases and a medicine group of 128 cases. The acupuncture group were treated by acupuncture at 9 acupoints such as Yamen (GV 15), Laogong (PC 8), Sanyinjiao (SP 6), etc. plus massage, and the medicine group by dong chuang Plaster. RESULTS: In the acupuncture group, 111 cases were cured and 136 cases were effective, the effective rate being 100.0%; and in the medicine group, 55 cases were cured and 98 cases were effective, the effective rate being 76.6%, with a very significant difference between the two groups in the therapeutic effect (P < 0.01). CONCLUSION: Acupuncture plus massage is superior to dong chuang plaster in therapeutic effect on chilblains. PMID- 16312924 TI - [Observation on therapeutic effect of acupuncture at lateral cervical Jiaji points on cervical spondylosis of vertebroarterial type]. AB - OBJECTIVE: To compare therapeutic effects of acupuncture at lateral cervical Jiaji and at traditional Jiaji points on cervical spondylosis of vertebroarterial type. METHODS: Seventy-two cases of vertebroarterial type cervical spondylosis were randomly divided into a needling lateral cervical Jiaji group (n=40) and a traditional Jiaji group (n=32). After 2 courses, therapeutic effects were evaluated and ultrasound Doppler's detection of skull was conducted. RESULTS: The therapeutic effect in the lateral cervical Jiaji group was better than that in the traditional Jiaji group (P < 0.05); And there was a significant or very significant difference between the two groups in the velocity of blood flow and the index of vascular pulsation (P < 0.05 or P < 0.01). CONCLUSION: The results of acupuncture at lateral cervical Jiaji points is better than that of acupuncture at traditional Jiaji points for treatment of cervical spondylosis of vertebroarterial type. PMID- 16312925 TI - [Clinical observation on electroacupuncture combined with massage for treatment of knee osteoarthritis complicated by gonarthromeningitis]. AB - OBJECTIVE: To observe therapeutic effect of acupuncture and moxibustion combined with massage on gonarthromeningitis complicated with knee osteoarthritis of Bengalese. METHODS: Acupuncture combined with electroacupuncture was given at Heding (Ex-LE2), Nei- and Wai-Xiyan (Ex-LE4, Ex-LE5) and others. and massage was given around the knee joints to treat 51 Bengalese patients of gonarthromeningitis complicated by knee osteoarthritis for one course (10 sessions of treatment). RESULTS: Seventeen cases were markedly-effective, 25 were effective, and 9 were ineffective, with a total effective rate of 82.4%. CONCLUSION: Chinese traditional acupuncture, moxibustion and massage have satisfactory therapeutic effect on gonarthromeningitis complicated by knee osteoarthritis of Bengalese. PMID- 16312926 TI - [Study on the needling depth of lumbar Jiaji (Ex-B2) points with CT imaging location]. AB - OBJECTIVE: To study on safe depth and angle of needling lumbar Jiaji (Ex-B2) for treatment of prolapse of lumbar intervertebral disc. METHODS: CT technique was used for scanning investigation on the depth and angle of needling lumbar Jiaji (Ex-B2). RESULTS: When the acupuncture needle or puncture needle was inserted at an angle of 20-30 degrees to the sagittal plane of the human body, the tip of needle could reached to extradural posterior space of the depth of lumbar Jiaji points (being the best inserting depth), in which catgut or medicine could be placed. CONCLUSION: Acupuncture or catgut stimulating the extradural posterior space at the depth of lumbar Jiaji is superior to the traditional needling method in treatment of prolapse of lumbar intervertebral disc. PMID- 16312927 TI - [Therapeutic effect of Zhuang medicine medicated thread moxibustion on asthma of lung deficiency type]. AB - OBJECTIVE: To observe effects of zhuang medicine medicated thread moxibustion on asthma of lung deficiency type. METHODS: Seventy-two asthma of lung deficiency type were randomly divided into Zhuang medicine medicated thread moxibustion group and an acupuncture-moxibustion control group. They were treated by moxibustion 1-2 times each day, for 21 days. The therapeutic effects, serum immunoglobulin (Ig) and complement-3 (C3) contents, pulmonary function, and symptoms and signs were investigated. RESULTS: There was no significant difference between the two groups in the total therapeutic effect, but the clinical control rate in the treatment group was higher than that in the control group. The Zhuang medicine medicated thread moxibustion could obviously improve cough, dyspnea, and respiratory function, increase levels of IgG and C3, and decrease level of IgE. CONCLUSION: Zhuang medicine medicated thread moxibustion has a better therapeutic effect on asthma of lung deficiency type, with safety. PMID- 16312928 TI - [Parabiosis theory and effective stimulating amount, reinforcing-reducing, therapeutic time and course of acupuncture]. AB - Parabiosis theory clearly expounds the change course of stimulatiom, namely, parabiotic equalization phase, parabiotic deregulation phase and parabiotic inhibition phase, and it can clearly explain that the effective stimulating amount is directly related to the reinforcing or reducing, and establishment of the therapeutic course. That is, the large effective stimulating amount is reducing method and the small effective stimulating amount is reinforcing method. In clinical application, for deficiency syndrome and chronic diseases, the effective intensity of acuti puncture should be smaller and the needle should be retained for 20-30 min, and for excess syndrome and acute disd eases, the effective stimulating intensity of acupuncture should be larger and the needle should be retained for longer time. In acupuncture treatment, acupoints should be grouped and used alternately, and the interval between therapeutic courses should be established. PMID- 16312929 TI - [Meridian regulation is a course of communicating and implicating of the informational system with the material system]. AB - OBJECTIVE: To approach to the intrinsic mechanism of meridian regulation. METHODS: Summarize and analyze the results of the author's studies and the development of other studies about life regulating and meridian at home and abroad. CONCLUSION: The regulating mechanism of meridians has two levels. The macroscopic regulation center is the brain, and the microscopic regulation base is the energy and informational current that depends on electromagnetic and chemical oscillation mainly. The meridian regulation is the process of the energy and informational system communicating and implicating with the material system. Neuroendocrine is the link of communicating and implicating, and Ca2+ oscillation frequencies exert intermediary function between the two systems. PMID- 16312930 TI - [Study on the law of circulation of meridians]. AB - OBJECTIVE: To study the basic law of circulation of channels and collaterals. METHODS: Inherit and develop ripe experiences of predecessors based on The Yellow Emperor's Internal Classic and other classic medical books. CONCLUSION: Circulation of channels and collaterals has the eight laws, including naming law, distribution law, converging law, exterior-interior association law, beginning ending running law, meridian-qi bidirectional circulation law, zang- and fu-organ pathway liaison law, and liaison law of connecting with trunks and sense organs. PMID- 16312931 TI - [Study on correlativity of different syndrome types with acupoint resistance in the patient of epigastralgia]. AB - OBJECTIVE: To study on correlativity of different syndrome types with acupoint resistance in the patient of epigastric pain. METHODS: Changes of electric resistance of acupoints in 60 cases of epigastric pain with different syndrome types were detected by TZ-01 model acupoint resistance determination instrument, and the correlativity was analyzed. RESULTS: Imbalance of resistance occurred at special acupoints of the spleen and stomach channels in all of the 60 patients of epigastric pain. The imbalance of resistance at Zusanli (ST 36) was the most obvious, and the response of the imbalance of resistance of acupoints was different among different syndrome types. CONCLUSION: There is correlativity of different syndrome types with changes of acupoint resistance in the patient of epigastric pain. PMID- 16312932 TI - [Thinking of application of gene chip technique in the study of channel-viscera correlativity]. AB - OBJECTIVE: To probe into thinking and prospect of application of gene chip technique to research of the channel-viscera correlativity. METHODS: Adopt literature analytic method, review and analyze current state of application of gene chip in biology and medicine, and achievements in studies of channel-viscera correlativity. RESULTS: In biological and medical fields, gene chip can be applied to high flux expression parallel analysis, large-scale gene discovery and gene analysis, gene polymorphous analysis, genome study, etc, especially, it has important application values in gene expression map; channel-viscera correlativity need stereo-crossing studies by multiple subjects, multiple systems, multiple directions and multiple levels, being more systematic and sequential, and gene chip technique is applicable to this requirement. CONCLUSION: Wide application of gene chip technique will comprehensively promote deep study of channel-viscera correlativity. PMID- 16312933 TI - [On the needling depth of filiform needle at acupoints]. AB - OBJECTIVE: To promote the study on standardization of needling depth of acupoints, ensure safety of needling, and promote internationally academic exchanges of acupuncture and moxibustion. METHODS: Study, analyze ancient and present literature of acupuncture and moxibustion, and compare the differences in needling depth, measure unit, description methods, and the causes and trend, and indicate confusion at present, and put forward a proposition for establishing standardization of needling depth. CONCLUSION: There are a lot of confusions in needling depth of acupoints from the ancient times to the present, which produces severe unfavorable influence on standardization and international exchanges of acupuncture and moxibustion science, and it remains to be studied for the standardization as early as possibly. PMID- 16312934 TI - [Study and clinical application of the book "A Study on The Eight Extra Channels"]. AB - Li Shi-zhen's contribution for eight extra-channel theory and effects of this theory on medicine, and the author's experiences on clinical application of this theory are introduced. It is held that the eight extra-channel theory, one of the essence of TCM, has wide application values in resolving clinical practical problems. PMID- 16312935 TI - [Effects of pricking blood therapy on ACTH contents in the hypothalamus-pituitary adrenal axis in the rat of experimental allergic rhinitis]. AB - OBJECTIVE: To explore the mechanism of pricking blood therapy (PBT) for allergic rhinitis (AR). METHODS: The model rat of experimental allergic rhinitis (EAR) was established by administration of ovoglohulin and they were divided into five groups randomly: a model group, a PBT group, a TCM point application group, a beclomethasone dipropionate (BDP) treatment group, and a health control group. Adrenocorticotrophic hormone (ACTH) contents in hypothalamus, pituitary gland, adrenal gland and plasma were determined by radioimmunoassay before and after treatment. RESULTS: The ACTH levels in the hypothalamus, pituitary gland, adrenal gland and plasma in the rat of EAR were lower than those in the health control group (P < 0.01). After treatment of pricking blood ther, ACTH levels in the above tissues increased significantly (P < 0.01), with very significant differences (P < 0.01) as compared with the model group, similar to those in the BDP group (P > 0.05). CONCLUSION: Pricking blood therapy has a regulatory action on ACTH in HPA axis of rats with experimental allergic rhinitis. PMID- 16312936 TI - [Effects of different point association needling methods on IL-1, sICAM-1 and sVCAM-1 in the patient of cerebral infarction]. AB - OBJECTIVE: To observe effects of acupuncture on serum interleukin-1 (IL-1) and soluble intercellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) in the patient of cerebral infarction. METHODS: Eighty-two cases of cerebral infarction were randomly divided into a treatment group treated by balanced muscular tension needling method, and a control group by traditional needling method. The levels of serum IL-1, sICAM-1 and sVCAM-1 were determined with enzyme-linked immunosorbent assay (ELISA). RESULTS: The levels of serum IL 1, sICAM-1 and sVCAM-1 in the patient of cerebral infarction were significantly higher than those in the healthy persons (P < 0.01); there were positive correlation between sICAM-1 or sVCAM-1 level and IL-1 level (r = 0.743, P < 0.001; r = 0.862, P < 0.001). The three indexes improved to a certain extent in the two groups, with the treatment group superior to the control group (P < 0.01). CONCLUSION: The improvement of nervous function by the balanced muscular tension needling method in the patient of cerebral infarction is possibly related with decrease of IL-1, sICAM-1 and sVCAM-1 levels. PMID- 16312937 TI - [Effects of electroacupuncture at different frequencies on morphological changes of nervous tissues and electromyogram of skeletal muscles in the rat with injury of sciatic nerve]. AB - OBJECTIVE: To observe effects of electroacupuncture of different frequencies on morphological changes of nervous tissues and electromyogram (EMG) of skeletal muscles in regeneration,so as to find more proper parameters of electroacupuncture. METHODS: The nerve regeneration chamber was built after the sciatic nerve was transected, and acupuncture was given at "Huantiao" (GB 30) , "Zusanli" (ST 36) and "Sanyinjiao" (SP 6) at the affected side with different frequencies (5 Hz, 100 Hz), 30 min each time,once every other day, 3 time each week, for 20 weeks and a control group was set up for natural recovery. The nerve fibers were stained by silver dying and ultrastructures of nerve tissues were observed. EMG of gastrocnemius was determined by Biopac physiologic apparatus. RESULTS: Electroacupuncture could promote the recovery of form of nerve tissues after the transection of sciatic nerve and functions of de-innervated muscle tissues after lesion of sciatic nerve, and the therapeutic effect of electroacupuncture at 5 Hz was the best. CONCLUSION: Electroacupuncture is an important way of promoting the regeneration of the lessoned peripheral nerve. PMID- 16312938 TI - [Advances in clinical and mechanism studies of acupuncture and moxibustion for treatment of depression]. AB - OBJECTIVE: To introduce clinical and mechanism studies about treatment of depression by acupuncture and moxibustion in recent years. METHODS: Review was made from clinical controls, mechanism studies and animal experiments. CONCLUSION: Acupuncture and moxibustion have definite therapeutic effect on depression, with less adverse effects, but some problems in clinical and mechanism studies remain to be resolved. PMID- 16312939 TI - A profile of chronic mental and physical conditions among African-American and Latino children in urban public housing. AB - OBJECTIVE: This study provides a profile of chronic mental and physical conditions among African-American and Latino children in urban public housing communities in Los Angeles, California. METHODS: The study focused on 187 African American and Latino households with children, 65% of a random sample of 287 households in three urban public housing communities. RESULTS: The findings suggest that minority children residing in public housing are one of the more severely health-compromised groups among under-served communities. Children of Latino and African-American families in our sample are two to four times more likely to suffer from chronic physical and mental conditions than the general population. The top five childhood chronic conditions reported by parents for one or more children in their households were asthma (32%), eye/vision problems (24%), dental problems (16%), Attention Deficit Hyperactivity Disorder (17%), and depression (8%). CONCLUSION: This study documents significant health disparities in this population and strongly suggests the need for future investigations in similar settings nationwide. PMID- 16312940 TI - Multi-cultural surveillance for ectopic pregnancy: California 1991-2000. AB - BACKGROUND: National surveillance for ectopic pregnancy (EP), the primary cause of maternal death in the first trimester, began in 1970. EP rates peeked during the late 1980s and have been highest for African Americans. However, limited reports on EP rates exist for other racial/ethnic groups. OBJECTIVES: To report state-level, multicultural trends in EP rates from 1991 to 2000. METHODS: Secondary data analysis of the California hospital discharge database collected by the Office of Statewide Health Planning and Development by using codes 633.0 633.9, from the International Classification of Diseases, Ninth Revision. EP rates are reported per 1000 pregnancies. RESULTS: From 1991 to 2000, 62,839 EP were reported in California. Mean EP rate was 11.2/1000, decreased from 15/1000 to 9.3/1000, and varied significantly by race/ethnicity and age. EP rates were highest among African Americans (25/1000) and lowest among Hispanics (7.7/1000); African Americans had higher odds of having EP relative to non-Hispanic Whites (odds ratio [OR] 2.14, 95% confidence interval [CI] 2.09-2.19). Women 35-44 years of age had the highest EP rates (17.6/1000) and higher odds of having EP compared to other age groups (OR 2.45, 95% CI 2.39-2.50). The highest rate of EP was found among African Americans 35-44 years of age (43.1/1000). CONCLUSIONS: The study showed declining EP rates in California for all groups but highlights disparities in EP rates and the attenuated rate of decline for African Americans, which places them at highest risk of death in the first trimester. This study demonstrates the usefulness of analyzing state-level data that may differ from aggregated national data when studying culturally diverse populations. PMID- 16312941 TI - Mortality trends of HIV-infected patients after the introduction of highly active antiretroviral therapy: analysis of a cohort of 3,322 HIV-infected persons. AB - INTRODUCTION: The implementation of highly active antiretroviral therapies (HAART) has reduced the mortality attributed to the human immunodeficiency virus (HIV) infection. Variation in the specific causes of death has also changed since the implementation of these therapies. METHODS: A prospective study was performed in 3322 HIV-infected persons enrolled in Puerto Rico between 1992 and 2003. We measured the mortality rates and the causes of death as listed in the death certificate and analyzed the variation as a function of the antiretroviral therapy (ART) use. Statistical analyses were performed to evaluate differences. RESULTS: The study found that persons treated with HAART had significantly lower mortality risk than ART-naive persons, regardless of gender and the use of injecting drugs. AIDS-defining conditions as a cause of demise were less frequently reported in patients with HAART. Gastrointestinal dysfunction, sepsis, metabolic abnormalities, and non-Kaposi neoplasms were more frequently reported as causes of death in patients treated with HAART. Hepatic failure as cause of death was also more frequent in these patients. The variation in the mortality trends was similar in both genders and according to the presence or absence of intravenous drug use. CONCLUSIONS: Highly active antiretroviral therapies (HAART) is associated with significant reduction in mortality and an increment in gastrointestinal dysfunction, sepsis, non-Kaposi neoplasms, and metabolic disorders as listed causes of death. Adverse and toxic profile of ART, along with the potential synergy of concomitant conditions, may accelerate these trends. Continued mortality surveillance of HIV/AIDS is imperative to follow the epidemic changes. PMID- 16312942 TI - Invasive group A streptococcal disease: race, hypotension, and immunogenetics. AB - OBJECTIVE: To determine if there was an association between race and the incidence of hypotension among patients hospitalized for invasive group A streptococcal disease (IGASD). DESIGN: Data were obtained from a retrospective cohort study of IGASD patients. The subjects were hospitalized throughout Florida for IGASD between 1996 and 2000 (N=151). The exposure variable for the current study was race (African Americans, Caucasians). The binary outcome was the presence of hypotension within 48 hours of hospital admission. A sensitivity analysis was performed to determine if a particular human leukocyte antigen (HLA) class II haplotype (DRB1*1501/DQB1*0602) confounded the association between race and the risk of hypotension. IGASD patients possessing this haplotype are less likely to experience hypotension with multiple organ failure. RESULTS: We found that Caucasians had three times the odds of developing hypotension than African Americans (adjusted odds ratio=3.02, 95% confidence interval: 1.10-8.29, P=.03). The sensitivity analysis revealed that the HLA class II haplotype DRB1*1501/DQB1*0602 was most likely not a confounder. CONCLUSIONS: Caucasians were significantly more likely than African Americans to develop the adverse outcome of hypotension within 48 hours of being hospitalized for IGASD. This excess risk is most likely not due to a particular host genetic factor, the HLA class II haplotype DRB1*1501/DQB1*0602. PMID- 16312943 TI - HIV infection and risk behavior of Hispanic farm workers at the west Texas-Mexico border. AB - This study investigated the risk behaviors and HIV infection rate in a sample of 210 migrant and seasonal farm workers (MSFW) working in the border city of El Paso, Texas and nearby communities. Surveys and structured interviews collected data on sexual behavior, drug use, condom use, and other potential risk behaviors associated with HIV infection. In addition, all subjects were tested for HIV exposure by using commercial kits. The MSFW participants were all Hispanic and comprised 156 males and 54 females. Only a small minority of the subjects reported engaging in same-sex (1.4%) or bisexual relations (2.8%). Most reported vaginal intercourse (94.7%), while 9% of males and 7.4% of females also reported anal intercourse. Forty-eight percent of the sample reported having sexual activity under the influence of alcohol (44%) and/or other drugs (14%). In this study, only 3.8% admitted to intravenous drug use. Furthermore, most reported that they never used any barrier method during vaginal (71.7%), anal (72.0%), or oral intercourse (87.5%). Only one subject, a male with multiple sex and needle sharing partners was HIV positive. Although a low level (.47%) of HIV infection was detected in the MSFW population tested, this rate is much higher than that reported for the rest of the county (.0099%) and indicates that this population is at a higher risk of HIV infection. PMID- 16312944 TI - Improving Hawaiian and Filipino involvement in clinical research opportunities: qualitative findings from Hawai'i. AB - OBJECTIVE: Investigate the barriers to participation in medical research that involves Asian and Pacific Islander (API) populations in Hawai'i. PARTICIPANTS: Fifty people (27 Filipinos, 23 Hawaiian/Pacific Islanders) in five different communities on Oahu. DESIGN: Nine focus groups with an ethnically matched moderator were held to explore people's feelings, problems, and recommendations regarding medical research. Sessions were audiotaped, transcribed, and qualitatively analyzed with the constant comparison method. RESULTS: Only 12% of study participants said that they absolutely would not participate in a clinical study. Most agreed that research is vital. Filipino participants were more optimistic about the safety and value of joining in medical research. Hawaiian groups were more hesitant and fearful. Reasons for nonparticipation included negative feelings about the purpose and intent of clinical trials and language and cultural barriers. Suggestions on how to encourage API populations to participate in research investigations included improving peoples' understanding of the benefits to family and community. Hawaiian and Filipino groups differed only slightly in their assessments of the type of research needed in their communities. CONCLUSIONS: Recruitment campaigns must improve people's awareness of the process of informed consent, research safeguards, and benefits to family and community. Attention should focus on K-12 health education to use members of the younger generations to access and educate elders, involving persons with medical research experience as a recruitment resource, returning results to study participants, and increasing the number of healthcare professionals and researchers that are culturally and linguistically matched to the community. PMID- 16312945 TI - A survey of endocrine disrupting chemicals in sewage and a preliminary treatment trial. AB - This paper demonstrates that synthetic endocrine disrupting chemicals (EDCs) (e.g. bisphenol-A) and natural EDCs (e.g. estrone and 17beta-estradiol) were found in the crude sewage from two sewage treatment works. Conventional biological processes can lower EDC concentrations to several hundred nanograms per litre. Because natural EDCs (e.g. estrone and 17beta1-estradiol) have biological activity and an adverse impact on the environment at extremely low concentrations (several tens of nanograms per litre), and because the existing wastewater/sewage treatment processes are not adequate to reduce natural EDC concentrations lower than 100 ng L(-1), further treatment after conventional biological processes is required. Preliminary trials with ferrate (VI) and electrochemical oxidation processes demonstrated that the former is more effective than the latter in reducing EDC concentrations, and that both the processes can effectively reduce EDCs to very low levels, ranging between 20 and 100 ng L(-1). PMID- 16312946 TI - Removal of pharmaceutically active compounds in nitrifying-denitrifying plants. AB - The behaviour of nine pharmaceutically active compounds (PhACs) of different diagnostic groups is studied during a nitrifying-denitrifying process in an activated sludge system. The compounds selected cover a wide range of frequently used substances such as anti-epileptics (carbamazepine), tranquillisers (diazepam), anti-depressants (fluoxetine and citalopram), anti-inflammatories (ibuprofen, naproxen and diclofenac) and estrogens (estradiol and ethinylestradiol). The main objective of this research is to investigate the effect of acclimation of biomass on the removal rates of these compounds, either by maintaining a high sludge retention time or at long-term operation. The removal rates achieved for nitrogen and carbon in the experimental unit exceed 90% and were not affected by the addition of PhACs. Carbamazepine, diazepam and diclofenac were only removed to a small extent. On the other hand, higher removal rates have been observed for naproxen and ibuprofen (68% and 82%), respectively. PMID- 16312947 TI - Resistance of Legionella to disinfection in hot water distribution systems. AB - The efficiency of various disinfection treatments against Legionella was tested on a hot water distribution system (HWDS) pilot unit. The results demonstrated clearly that most Legionella in the networks were fixed in the biofilm at the surface of the pipe (more than 98% for each loop). Chemical treatments (continuous chlorination, hyperchlorination, hydrogen peroxide and peracetic acid mixing) commonly used for the eradication of Legionella in hot water distribution networks appeared to be inadequate for eradicating the bacteria in the biofilm. Unfortunately, the biofilm contained most of the pathogens in an HWDS whereas legislation is only restricted to the Legionella concentration in the water phase. Thermal treatment appeared to be efficient to disinfect most of the biofilm but seemed to promote the biofilm re-growth as well. It was then concluded that the best solution to prevent Legionella contamination in hot water distribution systems would be to have perfect control of the temperature in the networks (temperature > 55 degrees C at all points). Nevertheless, in many cases it is difficult to have such control, so during the time necessary to modify networks, the best solution to control Legionella proliferation appears to be to apply a treatment shock (thermal or chlorination as a function of pipe characteristics). These treatments must be followed by a continuous chlorination that is totally controlled and equipped with alarm systems. This study demonstrates that biofilm sampling devices must be installed in hot water distribution systems to anticipate Legionella contamination and correctly determine the efficiency of the treatments. PMID- 16312948 TI - Behaviour of pharmaceuticals and personal care products in a sewage treatment plant of northwest Spain. AB - Thirteen pharmaceutical and cosmetic compounds have been surveyed along the different units of a municipal sewage treatment plant (STP) to study their fate across each step and the overall removal efficiency. The STP studied corresponds to a population of approximately 100,000 inhabitants located in Galicia (northwest Spain), including three main sections: pre-treatment (coarse and fine screening, grit and fat removal); primary treatment (sedimentation tanks); and secondary treatment (conventional activated sludge). Among all the substances considered (galaxolide, tonalide, carbamazepine, diazepam, diclofenac, ibuprofen, naproxen, estrone, estradiol, ethinylestradiol, roxitromycin, sulfamethoxazole and iopromide), only significant concentrations were found for two musks (galaxolide and tonalide), two antiphlogistics (ibuprofen and naproxen), two natural estrogens (estrone, estradiol), one antibiotic (sulfamethoxazole) and the X-ray contrast media (iopromide), being the other compounds below the quantification level. In the primary treatment, only the fragrances were partly removed, with efficiencies of 20-50% for galaxolide and tonalide. However, the aerobic treatment caused an important reduction in all compounds detected, between 35 and 75%, with the exception of iopromide. The overall removal efficiency of the STP ranged between 70 and 90% for the fragrances, 45 and 70% for the acidic compounds, around 67% for estradiol and 57% for the antibiotic sulfamethoxazole. PMID- 16312949 TI - Elution and adsorptive concentration of Japanese cedar (Cryptomeria japonica) pollen allergen in environmental water. AB - Elution of Japanese cedar pollen allergens (Cry j I and others) from pollen grains and its adsorptive concentration onto hydrophobic and hydrophilic surfaces were investigated using the surface plasmon resonance technique. Results showed that the allergen elution was obviously enhanced when the ion concentration was higher than that within the human body, indicating that the pollen tend to release its allergen in environmental water having a high ion concentration. However, higher adsorption capacity was observed on hydrophobic surface than hydrophilic surface. These results indicate that water puddles on roadsides beside heavy traffic including large amounts of ion compounds and hydrophobic diesel exhaust particles (DEPs) are a pollen allergen-DEP complex generator. DEPs are easily absorbed into the living body; therefore these mechanisms may be responsible for causing the highest incidence of pollinosis among residents living alongside roads with heavy automobile traffic. PMID- 16312950 TI - Degradation of parathion and the reduction of acute toxicity in TiO2 photocatalysis. AB - Photocatalytic degradation of methyl parathion was done using a circulating TiO2/UV and TiO2/solar reactor. Indoor experimental results showed that, under the photocatalysis conditions, parathion was more effectively degraded than under the photolysis and TiO2 only conditions. Parathion (38 microM) was completely degraded under photocatalysis within 90 min, and more than 80% TOC decrease after 150 minutes. The main ionic byproducts during the photocatalysis were measured, and almost complete nitrogen recovery was achieved as mainly NO3- NO2-, and NH4+, and 80% of sulfur as recovered as SO4(2)-. Organic intermediates such as nitrophenol and methyl paraoxon were also identified during the photocatalysis of parathion, and these were further degraded after 90 minutes. Microtox bioassay using Vibrio fischeri was used in evaluating the toxicity of solutions treated by photocatalysis and photolysis of parathion. The results showed that the acute toxicity expressed as EC50 almost reduced after 90 min under the photocatalysis condition whereas only 40% reduction of toxicity as EC50 was achieved in photolysis condition. The outdoor results using a TiO2/solar system were similar to the TiO2 indoor system, indicating the possibility of applying TiO2/solar system for the treatment of parathion-contaminated water. PMID- 16312951 TI - Analysis of drugs and personal care products in French source and drinking waters: the analytical challenge and examples of application. AB - This paper illustrates the challenge faced by analytical chemists when trying to measure selected compounds representative of various classes of prescription and hospital drugs. Because hundreds of drugs belonging to a wide variety of chemical groups are allowed for use, an array of analytical methods has to be implemented. As an example, as part of the European Poseidon Project, five different methods were required to measure eight drugs and personal care products. These methods are discussed in detail. Examples of application to surface and ground waters from the Paris area are also reported. The antibiotics roxithromycin and sulfamethoxazole were detected for the first time in the Seine River downstream of Paris. The behaviour of the eight target compounds during aquifer recharge and drinking water treatment is described. An incident involving the detection of micrograms per litre levels of the personal care product Galaxolide in a drinking water distribution system is reported. The value of the pharmaceuticals and personal care products selected as potential indicators is also discussed. PMID- 16312952 TI - Role of hydrophilic organic matter on developing toxicity in decay process of activated sludge. AB - It is known that the toxicity of effluent is more intensive than that of influent in the activated sludge process. In this study, we applied bioassay using cultured human cell lines to the decay process of activated sludge to evaluate the toxicity of organic matter generated and/or released from activated sludge bacteria. We also applied this bioassay to hydrophilic fraction of samples. The bioassay results showed that: (1) the variation in the dose-response relation obtained from assay with original samples was observed during decay; (2) on the other hand, the response curves of only hydrophilic fraction at each time show the same relationship between TOC and viability of MCF7 cells; (3) this trend was confirmed by plotting the time course of EC50. These results imply that: (1) the hydrophilic organic matter controlled for developing toxicity during decay process of activated sludge; and (2) the character of hydrophilic organic matter is not changed during the experimental period. PMID- 16312953 TI - Disinfection by-products and microbial contamination in the treatment of pool water with granular activated carbon. AB - For swimming pools, it is generally agreed that free chlorine levels have to be maintained to guarantee adequate disinfection. Recommended free chlorine levels can vary between 0.3 and 0.6 mg/L in Germany and up to 3 mg/L in other countries. Bathers introduce considerable amounts of organic matter, mainly in the form of such as urine and sweat, into the pool water. As a consequence, disinfection byproducts (DBPs) are formed. Regulations in Germany recommend levels of combined chlorine of less than 0.2 mg/L and levels of trihalomethanes (THMs) of less than 20 microg/L. Haloacetic acids (HAAs), haloacetonitriles (HANs), chloropicrin and chloral hydrate are also detected in considerable amounts. However, these compounds are not regulated yet. Swimming pool staff and swimmers, especially athletes, are primarily exposed to these byproducts by inhalation and/or dermal uptake. In Germany, new regulations for swimming pool water treatment generally require the use of activated carbon. In this project, three different types of granular activated carbon (GAC) (one standard GAC, two catalytic GACs) are compared for their long time behaviour in pool water treatment. In a pilot plant operated with real swimming pool water, production and removal of disinfection byproducts (THMs, HAAs, AOXs), of biodegradable substances (AOC), of bacteria (Pseudomonas aeruginosa, Legionella, coliforms, HPC) as well as the removal of chlorine and chloramines are monitored as function of GAC bed depth. Combined chlorine penetrates deeper in the filter bed than free chlorine does. However, both, free and combined chlorine removal efficiencies decrease over the time of filter operation. The decreases of removal efficiencies are also observed for parameters such as dissolved organic carbon, spectral absorption coefficient, adsorbable organic carbon and most of the disinfection byproducts. However, THMs, especially chloroform are produced in the filter bed. The GAC beds were contaminated microbially, especially with P. aeruginosa. The contamination was not removable by backwashing with chlorine concentrations up to 2 mg/l free chlorine. PMID- 16312954 TI - A single well push-pull test method for in situ determination of denitrification rates in a nitrate-contaminated groundwater aquifer. AB - In this study a single-well, "push- pull" test method is adapted for determination of in situ denitrification rates in groundwater aquifers. The rates of stepwise reduction of nitrate to nitrite, nitrous oxide, and molecular nitrogen were determined by performing a series of push-pull tests. The method consists of the controlled injection of a prepared test solution ("push") into an aquifer followed by the extraction of the test solution/ground water mixture ("pull") from the same location. The injected test solution consists of ground water containing a nonreactive tracer and one or more biologically reactive solutes. Reaction rate coefficients are computed from the mass of reactant consumed and/or product formed. A single Transport Test, one Biostimulation Test, and four Activity Tests were conducted for this study. Transport tests are conducted to evaluate the mobility of solutes used in subsequent tests. These included bromide (a conservative tracer), fumarate (a carbon and/or source), and nitrate (an electron acceptor). Extraction phase breakthrough curves for all solutes were similar, indicating apparent conservative transport of the solutes prior to biostimulation. Biostimulation tests were conducted to stimulate the activity of indigenous heterotrophic denitrifying microorganisms and consisted of injection of site ground water containing fumarate and nitrate. Biostimulation was detected by the simultaneous production of carbon dioxide and nitrite after each injection. Activity tests were conducted to quantify rates of nitrate, nitrite, and nitrous oxide reduction. Estimated zero-order degradation rates decreased in the order nitrate > nitrite > nitrous oxide. The series of push-pull tests developed and field tested in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ denitrification in nitrate contaminated aquifers. PMID- 16312955 TI - Arsenic contamination of groundwater in Nawabganj, Bangladesh, focusing on the relationship with other metals and ions. AB - Serious arsenic contamination of groundwater in Bangladesh has been frequently reported and is of great concern. In this research, repeated water sampling from the same 10 tubewells in Nawabganj municipality, Bangladesh, was conducted and analysed, focusing on the seasonal variation of water quality and the relationship among arsenic and other metals and ions. For the seasonal variation of water quality, arsenic and iron concentrations were higher in the rainy season in general although the tendency was not consistent and it depended on the tubewell and the time. Correlation between arsenic and iron could not be observed in this study (r = -0.01) when using all cases. This was because no correlation was observed in the higher arsenic concentration range. Arsenic removal by co precipitation with coexisting iron is known as one of the locally applicable techniques in Bangladesh, but the result from this study suggests that some additional treatments such as the extra injection of iron should be performed in some cases, especially where the arsenic concentration is high. The correlation between arsenic and other substances was also analysed. As a result, manganese (r = 0.37), molybdenum (r = 0.33) and sulfate ion (r = -0.33) significantly correlated with arsenic (p < 0.05). The negative correlation between arsenic and sulfate ion implies the dissolution of arsenic into groundwater under reductive conditions although there are some exceptional cases. PMID- 16312956 TI - Effects of aging on the bioavailability and sorption/desorption behavior of biphenyl in soils. AB - The effects of aging (soil-chemical contact time) on bioavailability, one of the main variables for determining the persistence of organic chemicals in the environment, is poorly understood. There are few studies that have tested the effects of long-term aging on chemical dynamics in soils and have related these effects to bioavailability tests of these aged compounds. In this study, sorption/desorption behavior of biphenyl (BP) on two soil types (Capac A and Capac B) was evaluated for aging times of 24 hours and 8 months. Then bioavailability experiments of BP on the same soils were performed after aging times of 24 hours to 6 months. Sorption isotherms and desorption kinetic profiles were prepared to analyze changes in uptake and release, respectively, due to aging of BP. Mineralization kinetics of BP to 14CO2 by a strain of Pseudomonas putida was used to assess changes in bioavailability due to aging. Data indicates that there was an increase in sorption with aging time for BP on both soil types. The rate of BP desorption did not much change with increased aging time. The extent of BP mineralization was found to decrease with aging time. PMID- 16312957 TI - Effect of soil and a nonionic surfactant on BTE-oX and MTBE biodegradation kinetics. AB - The biodegradation kinetics of BTE-oX and MTBE, mixed all together, in the presence of 905 mg/L VSS of BTEX-acclimated biomass was evaluated. Effects of soil and Tergitol NP-10 in aqueous samples on substrate biodegradation rates were also evaluated. Biodegradation kinetics was evaluated for 36 hours, every 6 hours. MTBE biodegradation followed a first-order one-phase kinetic model in all samples, whereas benzene, toluene and ethylbenzene biodegradation followed a first-order two-phase kinetic model in all samples. O-xylene biodegradation followed a first-order two-phase kinetic model in the presence of biomass only. Interestingly, o-xylene biodegradation was able to switch to a first-order one phase kinetic model when either soil or soil and Tergitol NP-10 were added. The presence of soil in aqueous samples retarded benzene, toluene and ethylbenzene removal rates. O-xylene and MTBE removal rates were enhanced by soil. The addition of Tergitol NP-10 to aqueous samples containing soil had a positive effect on substrate removal rate in all samples. Substrate percent removals ranged 77-99.8% for benzene, toluene and ethylbenzene. O-xylene and MTBE percent removals ranged 50.1-65.3% and 9.9-43.0%, respectively. PMID- 16312958 TI - The rapid detection of methyl tert-butyl ether (MtBE) in water using a prototype gas sensor system. AB - The gasoline additive Methyl-tertiary-Butyl Ether (MtBE) is the second most common contaminant of groundwater in the USA and represents an important soil contaminant. This compound has been detected in the groundwater in at least 27 states as a result of leaking underground storage facilities (gasoline storage tanks and pipelines). Since the health effects of MtBE are unclear the potential threat to drinking water supplies is serious. Therefore, the ability to detect MtBE at low levels (ppb) and on-line at high-risk groundwater sites would be highly desirable. This paper reports the use of 'commercial' and metal oxide sensor arrays for the detection of MtBE in drinking and surface waters at low ppb level (microg.L(-1) range). The output responses from some of the sensors were found to correlate well with MtBE concentrations under laboratory conditions. PMID- 16312959 TI - Cometabolic transformation of cis-1,2-dichloroethylene and cis-1,2 dichloroethylene epoxide by a butane-grown mixed culture. AB - Aerobic cometabolism of cis-1,2-dichloroethylene (c-DCE) by a butane-grown mixed culture was evaluated in batch kinetic tests. The transformation of c-DCE resulted in the coincident generation of c-DCE epoxide. Chloride release studies showed approximately 75% oxidative dechlorination of c-DCE. Mass spectrometry confirmed the presence of a compound with mass-to-charge-fragment ratios of 112, 83, 48, and 35. These values are in agreement with the spectra of chemically synthesized c-DCE epoxide. The transformation of c-DCE required O2, was inhibited by butane and was inactivated by acetylene (a known monooxygenase inactivator), indicating that a butane monooxygenase enzyme was likely involved in the transformation of c-DCE. This study showed c-DCE epoxide was biologically transformed, likely by a butane monooxygenase enzyme. c-DCE epoxide transformation was inhibited by both acetylene and c-DCE indicating a monooxygenase enzyme was involved. The epoxide transformation was also stopped when mercuric chloride (HgCl2) was added as a biological inhibitor, further support a biological transformation. To our knowledge this is the first report of the biological transform c-DCE epoxide by a butane-grown culture. PMID- 16312960 TI - Water, sanitation and hygiene in developing countries: interventions and diarrhoea--a review. AB - We conducted a search to identify all English language papers (published between 1 January 1985 and 26 June 2003) with evidence on the effectiveness of water, sanitation and hygiene interventions in developing countries, in which diarrhoea morbidity in non-outbreak conditions was reported. A total of 39 studies were identified as relevant after an initial review of over 2000 titles. Data were extracted and, where possible, combined using meta-analysis to provide a summary estimate of the effectiveness of specific interventions, including water supply and water treatment. Most of the interventions (including multiple interventions, hygiene and water quality) were found to significantly reduce the levels of diarrhoeal illness, with the greatest impact being seen for hygiene and household treatment interventions (after removal of studies classed as poor quality). Sanitation interventions could not be assessed as only a single study suitable for meta-analysis was identified. PMID- 16312961 TI - Perception of tap water risks and quality: a structural equation model approach. AB - The present study aims to test a model of tap water risk and quality perception. A questionnaire was designed and applied to a convenience sample of 499 people in Portugal. The model includes aesthetic variables (colour, odour, and flavour), contextual indicators and risk perception. Other variables were also considered, including external information, trust in water companies, familiarity with tap water, and past water-related health problems. A behavioural outcome--drinking tap water--was also studied. Due to multivariate non-normality (Mardia's Coefficient (G2,P) = 460.3), the AGLS estimator was used for the structural equation model (SEM). A triangulation approach with multivariate regression analysis was used to explain the use of tap water to drink and to propose a more holistic model that could not be tested using SEM. Results show that perceived water quality is largely influenced by flavour. Other factors, such as perceived risk and contextual indicators, also have a role but their relevance is relatively weak. On the other hand, risk perception seems to be mainly a result of external information, past health problems, and water colour. Finally, the use of tap water to drink at home can be moderately explained and depends mostly on the ability to use alternatives (i.e. bottled water), tap water taste, and perceived risk. PMID- 16312962 TI - Excess propagation and biological control of zooplanktonic Cyclops in drinking water sources. AB - Starting with the occurrence of Cyclops in drinking water sources and its danger to drinking water safety, we extensively investigated the nature of its excess propagation by analyzing the effect of living space and predator pressure in the food chain on its survival. Then we developed an ecological project using fish biomanipulation, different to the conventional fishery culture technique, to control excess propagation of Cyclops. The control effects on Cyclops of four species of fish were investigated experimentally at a stocking density of 30 g/m3 of water. Results showed that the food habit of the fish had a significant influence on the biological control of Cyclops. The propagation of Cyclops could be controlled effectively and also the water quality improved simultaneously by stocking filter-feeding fish, such as silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis). However, herbivorous Ctenopharyngodon idellus and omnivorous Cyprinus carpio had no obvious biological effects on controlling the growth of Cyclops and restoring water quality. The results further proved that under conditions of proper poly-culture densities of silver carp and bighead carp, the number of Cyclops might be controlled at very low levels and eutrophication might be abated by removing nutrients from the water body. PMID- 16312963 TI - Environmental impact and health risks associated with greywater irrigation: a case study. AB - There is an increasing trend to use greywater for irrigation in households. This is partly due to the notion that greywater is of better quality than wastewater and therefore does not need extensive treatment beyond addressing public health issues. The aim of the study was to evaluate the environmental impact and health risks associated with the use of greywater for irrigation on a small private farm. Over a three-year period, each of three plots on a farm was irrigated with either freshwater, fertilized water, or greywater. Irrigation water and soil from the plots were analyzed for a wide range of chemical and microbial variables. Results suggest that greywater may be of similar quality to wastewater in several parameters such as BOD and faecal coliforms. For some other variables such as boron and surfactants, greywater may even be of worse quality than wastewater. Long-term irrigation of arid loess soil with greywater may result in accumulation of salts, surfactants and boron in the soil, causing changes in soil properties and toxicity to plants. Faecal coliforms did not survive in the soil. Treating greywater before using it for irrigation is recommended, even in places where this is not a requirement. PMID- 16312964 TI - Significance of bacteria associated with invertebrates in drinking water distribution networks. AB - The implication of invertebrates found in drinking water distribution networks to public health is of concern to water utilities. Previous studies have shown that the bacteria associated with the invertebrates could be potentially pathogenic to humans. This study investigated the level and identity of bacteria commonly associated with invertebrates collected from the drinking water treatment systems as well as from the main pipelines leaving the treatment works. On all sampling occasions bacteria were isolated from the invertebrate samples collected. The highest bacterial counts were observed for the samples taken before filtration as was expected. There were, however, indications that optimal removal of invertebrates from water did not always occur. During the investigation, 116 colonies were sampled for further identification. The isolates represent several bacterial genera and species that are pathogenic or opportunistic pathogens of humans. Diarrhoea, meningitis, septicaemia and skin infections are among the diseases associated with these organisms. The estimated number of bacteria that could be associated with a single invertebrate (as based on average invertebrate numbers) could range from 10 to 4000 bacteria per organism. It can, therefore, be concluded that bacteria associated with invertebrates might under the worst case scenario pose a potential health risk to water users. In the light of the above findings it is clear that invertebrates in drinking water should be controlled at levels as low as technically and economically feasible. PMID- 16312965 TI - Detection and identification of mycobacteria in the Lisbon water distribution system. AB - Mycobacteria have emerged as a major cause of opportunistic infections. Until the present, only a few studies have characterized mycobacteria present in the water distribution system of urban areas. In this study, we characterize these microorganisms in the Lisbon water distribution system. Our results indicate a high rate of positivities (90.5%) with mainly saprophytic mycobacteria. Around 63% of these results belong to strains of Mycobacterium gordonae indicating a generalized proliferation of this species in the Lisbon water distribution system. A total of 21.05% of the isolates are from M. kansasii, M. intracellulare and M. chelonae. PMID- 16312966 TI - Biofilms in an urban water distribution system: measurement of biofilm biomass, pathogens and pathogen persistence within the Greater Stockholm Area, Sweden. AB - Distribution pipe biofilms can provide sites for the concentration of a wide range of microbial pathogens, thereby acting as a potential source of continual microbial exposure and furthermore can affect the aesthetic quality of water. In a joint project between Stockholm Water, the MISTRA "Sustainable Urban Water" program, the Swedish Institute for Infectious Disease Control and the Royal Technical University, Stockholm, the aim of the current study was to investigate biofilms formed in an urban water distribution system, and quantify the impact of such biofilms on potential pathogen accumulation and persistence within the Greater Stockholm Area, Sweden. When used for primary disinfection, ultra-violet (UV) treatment had no measurable influence on biofilm formation within the distribution system when compared to conventional chlorination. Biofilms produced within a model pilot-plant were found to be representative to those that had formed within the larger municipal water distribution system, demonstrating the applicability of the novel pilot-plant for future studies. Polystyrene microspheres (1.0 microm) and Salmonella bacteriophages demonstrated their ability to accumulate and persist within the model pilot-plant system, where the means of primary disinfection (UV-treatment, chlorination) had no influence on such phenomena. With the exception of aeromonads, potential pathogens and faecal indicators could not be detected within biofilms from the Stockholm water distribution system. Results from this investigation may provide information for water treatment and distribution management strategies, and fill key data gaps that presently hinder the refinement of microbial risk models. PMID- 16312967 TI - A deterministic model to quantify pathogen loads in drinking water catchments: pathogen budget for the Wingecarribee. AB - This paper describes the development and testing of a mathematical model as a tool to quantify pathogen loads in Sydney's drinking water catchments. It has been used to identify, quantify and prioritise sources of Cryptosporidium, Giardia and E. coli in the Wingecarribee catchment. The pathogen model promotes understanding of the relative significance of different sources of pathogen risks as well as their fate and transport as they move through the subcatchments. This pathogen model not only enables water utility managers to identify those catchment segments that may contribute the highest load of pathogens, but also where management options will be most effective. PMID- 16312968 TI - Evidence for the existence of Cryptosporidium oocysts as single entities in surface runoff. AB - There is uncertainty whether Cryptosporidium oocysts attach to particles or to each other under ambient water conditions. Particle size distributions of Cryptosporidium oocyst suspensions were determined over a range of ionic strengths and pHs to determine under those environmental conditions that may promote oocyst aggregation. Cryptosporidium oocysts were shown to only aggregate in high ionic strength solutions (>0.45 M) and remain largely as single entities at ionic strengths and pHs that were likely to be encountered in surface runoff. Similarly, in loam soil suspensions, rather than attaching to the soil particles the majority of oocysts also remained as single entities. Overall, oocysts are expected to remain largely unattached to either themselves or soil particles in overland runoff. This has implications for pathogen transport and modelling since oocysts that are freely suspended are more likely to be transported in runoff to surface waters than if attached to more dense soil/faecal particles. PMID- 16312969 TI - Application of a virological tracer method for the assessment of pathogen removal by physicochemical treatment and chemical disinfection. AB - Coxsackie B3 (CoxB3) virus was used as a virological tracer for an assessment of the efficiency of pathogen removal by several typical physicochemical treatment and chemical disinfection processes, such as coagulation-filtration, ultra filtration, and disinfection using chlorine and ozone, with regard to the pathogenic quality of the treated domestic wastewater for reuse purposes. The CoxB3 virus was seeded to sterilized secondary effluent to make a raw water of known pathogenic level. After applying the raw water to each treatment or disinfection process, the residual virus in the finished water was concentrated, and virus assay was carried out by the Tissue Culture Infectious Dose technique. TCID50 was used as an indicative parameter of CoxB3 virus in the raw and treated water. Parallel experiments were also conducted to evaluate the effectiveness of each process for the removal of coliform bacteria. It was noticed from the experiment that both coagulation-filtration and ultrafiltration could achieve substantial removal of TCID50 at about the same level (2-log removal). However, the effect of the two processes on the removal of coliform bacteria was much different: 2-log removal by coagulation-filtration and 4 to 5-log removal by ultrafiltration. The TCID50 removal correlates more closely with the removal of turbidity than that of coliform bacteria. Chlorine was found to be effective in coliform removal but almost had no effect on TCID50. As ozone was applied, a high removal of both coliform bacteria and TCID50 could be obtained. PMID- 16312970 TI - Concentration and detection of SARS coronavirus in sewage from Xiao Tang Shan Hospital and the 309th Hospital of the Chinese People's Liberation Army. AB - A worldwide outbreak of severe acute respiratory syndrome (SARS) had been reported. Over 8439 SARS cases and 812 SARS-related deaths were reported to the World Health Organization from 32 countries around the world up to 5 July 2003. The mechanism of transmission of SARS-CoV has been limited only to close contacts with patients. Attention was focused on possible transmission by the sewage system because laboratory studies showed that patients excreted coronavirus RNA in their stools in Amoy Gardens in Hong Kong. To explore whether the stool of SARS patients or the sewage containing the stool of patients would transmit SARS CoV or not, we used a style of electropositive filter media particle to concentrate the SARS-CoV from the sewage of two hospitals receiving SARS patients in Beijing, as well as cell culture, semi-nested RT-PCR and sequencing of genes to detect and identify the viruses from sewage. There was no live SARS-CoV detected in the sewage in these assays. The nucleic acid of SARS-CoV was found in the sewage before disinfection from both hospitals by PCR. After disinfection, SARS-CoV RNA could be detected from some samples from the 309th Hospital of the Chinese People's Liberation Army, but not from Xiao Tang Shan Hospital after disinfection. In this study, we found that the virus can survive for 14 days in sewage at 4 degrees C, 2 days at 20 degrees C, and its RNA can be detected for 8 days though the virus had been inactivated. In conclusion, this study demonstrates that the RNA of SARS-CoV could be detected from the concentrates of sewage of both hospitals receiving SARS patients before disinfection and occasionally after disinfection though there was no live SARS-CoV; thus much attention should be paid to the treatment of stools of patients and the sewage of hospitals receiving SARS patients. PMID- 16312971 TI - Assessment of solar photocatalysis to purify on-site rinse waters from tractor cisterns used in grapevine pest control: field experimentation. AB - The aim of this study was to assess in a vineyard the effect of purifying by solar photocatalysis the title rinse waters (currently most often rejected) in terms of efficacy and on-site practicality for the wine grower. The on-site, self functioning, solar purifying unit included a corrugated-steel inclined plate of area S = 1 m2 onto which a TiO2-coated thin material had been slightly pressed, a tank, and an aquarium-type pump powered by a photovoltaic panel (appropriate for isolated locations). For a vineyard of area A = 0.15 km2, the rinse water (about 90 L) corresponding to each of four typical vine treatments in summer was analysed (major pesticides for each treatment, TOC, Microtox test and, in one case, BOD5) by independent laboratories, before and after purification for 8 days. The S/A ratio tested was found insufficient even if the photocatalytic treatment markedly improved the quality of the rinse waters. From the relatively low final organic content reached in one case, it is calculated that a three-time higher S/A ratio might suffice, but new trials are necessary to determine whether it is valid for other typical cases. Inferred contribution of inorganic ions to the post-photocatalytic treatment toxicity points to the need for an additional detoxification. These field experiments have also demonstrated that the purifying prototype is robust, and easy to install and use on site by the wine grower. PMID- 16312972 TI - Evaluation of heavy metal inhibition of activated sludge by TTC and INT-electron transport system activity tests. AB - TTC and INT-electron transport system activity tests were compared for assessing heavy metal inhibition of activated sludge. The median inhibitory concentrations (IC50s) of Cu2+, Zn2+, Cd2+, Hg2+, Ni2+, Pb2+ and Ag+ measured via TTC test were lower than those measured via INT test, which indicates that the INT-electron transport system activity test was less sensitive to heavy metal toxicity than the TTC test. Tested heavy metals brought about similar decrease in TTC-electron transport system activity and COD removal rate, but less decrease in INT-electron transport system activity than COD removal rate, which suggests that the TTC electron transport system activity is a better parameter for reflecting heavy metal inhibition of activated sludge than INT-electron transport system activity. The ranking of tested heavy metals in order of decreasing toxicity based on TTC test was Hg2+, Cd2+, Cu2+, Ag+, Zn2+, Ni2+ and Pb2+, and the ranking based on INT test was Hg2+, Ag+, Cu2+, Cd2+, Zn2+, Ni2+ and Pb2+. PMID- 16312973 TI - Biochemical pathway and degradation of phthalate ester isomers by bacteria. AB - Degradation of dimethyl isophthalate (DMI) and dimethyl phthalate ester (DMPE) was investigated using microorganisms isolated from mangrove sediment of Hong Kong Mai Po Nature Reserve. One enrichment culture was capable of utilizing DMI as the sole source of carbon and energy, but none of the bacteria in the enrichment culture was capable of degrading DMI alone. In co-culture of two bacteria, degradation was observed proceeding through monomethyl isophthalate (MMI) ester and isophthalic acid (IPA) before the aromatic ring opening. Using DMI as the sole carbon and energy source, Klebsiella oxytoca Sc and Methylobacterium mesophilicum Sr degraded DMI through the biochemical cooperation. The initial hydrolytic reaction of the ester bond was by K. oxytoca Sc and the next step of transformation was by M. mesophilicum Sr, and IPA was degraded by both of them. In another investigation, a novel bacterium, strain MPsc, was isolated for degradation of dimethyl phthalate ester (DMPE) also from the mangrove sediment. On the basis of phenotypic, biochemical and 16S rDNA gene sequence analyses, the strain MPsc should be considered as a new bacterium at the genus level (8% differences). This strain, together with a Rhodococcus zopfii isolated from the same mangrove sediment, was able to degrade DMPE aerobically. The consortium consisting of the two species degraded 450 mg/l DMPE within 3 days as the sole source of carbon and energy, but none of the individual species alone was able to transform DMPE. Furthermore, the biochemical degradation pathway proceeded through monomethyl phthalate (MMP), phthalic acid (PA) and then protocatechuate before aromatic ring cleavage. Our results suggest that degradation of complex organic compounds including DMI and DMPE may be carried out by several members of microorganisms working together in the natural environments. PMID- 16312974 TI - Dinitrogen oxide detection for nitrification failure early warning systems. AB - Experiments were conducted in order to establish whether N2O could be used to predict nitrification failure (through non-invasive means). Previous research had shown a strong correlation between N2O gas and NH3 in the effluent, giving rise to the possibility N2O can be used as an indicator for failure in the nitrification process. Two pilot-scale activated sludge plants were used, each with two lanes. The smaller consisted of a 601 aeration tank and a 201 clarifier; the larger pilot plant had an aeration tank of 3151 and a clarifier of 1001. The small pilot plant experiments showed that N2O gas was given off almost immediately from O2 deprivation/NH3 shock loads, but did not follow the expected trend of the time lag of NH3 in the effluent. This led to further investigation in the hydrodynamics and mixing characteristics of aeration basins, where a second larger pilot plant was used. Further experiments were conducted of high NH3 loadings and O2 deprivation, showed that work of was reproducible. However, it was also shown that with partial nitrification failure, a different N2O response of a continual rise was observed. PMID- 16312975 TI - Estimation of biodegradation kinetic constants for aromatic derivates through aerobic batch experiments. AB - The kinetics of aerobic biodegradation were studied for 20 aromatic species by using sludges taken from a municipal sewage treatment plant. The reproducibility of the results is tested with respect to the period of collection of the sludges and the wastewater treatment plant where they were taken. The comparison of kinetic constants estimated for investigated chemicals allows evaluation of the effect on the reactivity due to the presence of single groups (i.e. -OH, -CH3, Cl, -NO2) into the aromatic structures. The search for easy structure-reactivity relations is also attempted by using some group contributing methods. PMID- 16312976 TI - Aerobic phenanthrene biodegradation in a two-phase partitioning bioreactor. AB - The aerobic degradation of phenanthrene by a Pseudomonas migulae strain under classical mechanical aeration and under photosynthetic oxygenation (using a Chlorella sorokiniana strain) in a two-phase partitioning bioreactor (TPPB) constructed with silicone oil as organic phase was investigated. When traditional mechanical aeration was used, an increase in the aeration and/or in the agitation rate enhanced phenanthrene biodegradation. Thus, phenanthrene removal rates (based on the total liquid volume of cultivation) ranged from 22 +/- 1 to 36 +/- 2 mg/l h at 100 rpm and 1 vvm and 400 rpm and 3 vvm, respectively. On the other hand, during phenanthrene biodegradation using the algal-bacterial microcosm a maximum rate of 8.1 +/- 1.2 mg/l h at 200 rpm and 8000 lux of illuminance was achieved. PMID- 16312977 TI - Steroid estrogens in primary and tertiary wastewater treatment plants. AB - The concentrations of two natural estrogens (estrone (E1) and Estradiol (E2)) and one synthetic progestin (Ethinylestradiol (EE2)) were measured for different unit operations in an advanced sewage treatment plant and in a large coastal enhanced primary sewage treatment plant. The average influent concentration to both plants was similar: 55 and 53 ng/L for E1 and 22 and 12 ng/L for E2 for the advanced and enhanced primary STPs, respectively. The activated sludge process at the advanced STP removed up to 85% and 96% of E1 and E2, respectively. The enhanced primary sewage treatment plant was mostly ineffective at removing the steroids with only 14% of E1 and 5% of E2 being removed during the treatment process. EE2 was not been detected during the study period in the influent or effluent of either STP. The difference in the observed removal between the two plants is primarily linked to plant performance but the extent to which removal of steroid estrogens is due to bacterial metabolism (i.e. the advanced STP) rather than adsorption to the bacterial biomass remains unclear. The poor removal observed for the coastal enhanced primary STP may have implications for the receiving environment in terms of a greater potential for abnormal reproductive systems in marine animals, particularly if discharges are into large bays or harbours where flushing is limited. PMID- 16312978 TI - Predicting fate of the contraceptive pill in wastewater treatment and discharge. AB - The risk of endocrine disrupters to humans and wildlife is to date poorly understood, although evidence of effects is now widespread. In understanding the risk, an important step is the determination of the partitioning, as well as chemical and biochemical transformation, of compounds in the environment, the water cycle and the food chain. This is a complex task and this paper is a first step towards estimating some of these factors from a largely theoretical approach. A chemical fate model is used to predict the fate of the contraceptive drug 17alpha-ethinylestradiol (EE2). The example of the contraceptive pill is chosen to follow the journey of the drug from human ingestion and excretion to treatment in a sewage treatment plant (STP) using fugacity-based fate models, followed by discharge into a receiving river and eventually into the estuary/sea. The model predicts how EE2 will partition into the different compartments during each stage of this journey and thereby infiltrate into the food chain. The results suggest that a person would have to ingest more than 30,000 portions of fish to consume an equivalent to a single average dose of the contraceptive pill. While this scenario is highly unlikely, the biochemical consequence of the contraceptive pill is greatly significant. Furthermore, there are many identified similarly estrogenic compounds in the environment while this study only considers one. Cumulative effects of such compounds as well as degradation into other potent compounds may be anticipated. An important message in this paper is the interrelation of wastewater effluent discharge and eventual human exposure of marginally degradable and lipophilic chemicals. While at present the main concerns regarding endocrine disrupters appear to be the fear of their occurrence in drinking water sources, it is clear that the domains of wastewater treatment and discharge, water supply and contamination of food should not be treated as separate issues. The model suggests that exposure from food (contaminated by effluent) may be much more significant than from drinking water. PMID- 16312979 TI - The future of SC's malpractice system. PMID- 16312980 TI - A century of otolarngyology-head and neck surgery as revealed in The Journal of the South Carolina Medical Association. PMID- 16312981 TI - 100 years of psychiatry and neurology in The Journal of the South Carolina Medical Association. PMID- 16312982 TI - General surgery as reported in The Journal of the South Carolina Medical Association: a review from 1904-1954. PMID- 16312983 TI - General surgery as reported in The Journal of the South Carolina Medical Association: a review from 1955 to 2005. PMID- 16312984 TI - One hundred years of infectious diseases as reported in The Journal of the South Carolina Medical Association. PMID- 16312985 TI - Medical education in South Carolina, 1905-2004, as reflected in The Journal of the South Carolina Medical Association. AB - Medical education has from meager beginnings prospered in South Carolina through the past century, with most of the key advances being captured in The Journal of the South Carolina Medical Association. PMID- 16312986 TI - The Journal of the South Carolina Medical Association through the past century (part II). PMID- 16312987 TI - Small steps to take on obesity. PMID- 16312988 TI - Soil resource availability impacts microbial response to organic carbon and inorganic nitrogen inputs. AB - Impacts of newly added organic carbon(C) and inorganic nitrogen (N) on the microbial utilization of soil organic matter are important in determining the future C balance of terrestrial ecosystems. We examined microbial responses to cellulose and ammonium nitrate additions in three soils with very different C and N availability. These soils included an organic soil (14.2% total organic C, with extremely high extractable N and low labile C), a forest soil (4.7% total organic C, with high labile C and extremely low extractable N), and a grassland soil (1.6% total organic C, with low extractable N and labile C). While cellulose addition alone significantly enhanced microbial respiration and biomass C and N in the organic and grassland soils, it accelerated only the microbial respiration in the highly-N limited forest soil. These results indicated that when N was not limited, C addition enhanced soil respiration by stimulating both microbial growth and their metabolic activity. New C inputs lead to elevated C release in all three soils, and the magnitude of the enhancement was higher in the organic and grassland soils than the forest soil. The addition of cellulose plus N to the forest and grassland soils initially increased the microbial biomass and respiration rates, but decreased the rates as time progressed. Compared to cellulose addition alone, cellulose plus N additions increased the total C released in the grassland soil, but not in the forest soil. The enhancement of total C-released induced by C and N addition was less than 50% of the added-C in the forest soil after 96 d of incubation, in contrast to 87.5% and 89.0% in the organic and grassland soils. These results indicate that indigenous soil C and N availability substantially impacts the allocation of organic C for microbial biomass growth and/or respiration, potentially regulating the turnover rates of the new organic C inputs. PMID- 16312989 TI - Effects of phosphate on the adsorption of glyphosate on three different types of Chinese soils. AB - Glyphosate (GPS) is a non-selective, post-mergence herbicide that is widely used throughout the world. Due to the similar molecular structures of glyphosate and phosphate, adsorption of glyphosate on soil is easily affected by coexisting phosphate, especially when phosphate is applied at a significant rate in farmland. This paper studied the effects of phosphate on the adsorption of glyphosate on three different types of Chinese soils including two variable charge soils and one permanent charge soil. The results indicated that Freundlich equations used to simulate glyphosate adsorption isotherms gave high correlation coefficients (0.990-0.998) with K values of 2751, 2451 and 166 for the zhuanhong soil(ZH soil, Laterite), red soil(RS, Udic Ferrisol) and Wushan paddy soil (WS soil, Anthrosol), respectively. The more the soil iron and aluminum oxides and clay contained, the more glyphosate adsorbed. The presence of phosphate significantly decreased the adsorption of glyphosate to the soils by competing with glyphosate for adsorption sites of soils. Meanwhile, the effects of phosphate on adsorption of glyphosate on the two variable charge soils were more significant than that on the permanent charge soil. When phosphate and glyphosate were added in the soils in different orders, the adsorption quantities of glyphosate on the soils were different, which followed GPS-soil > GPS-P-soil = GPS-soil-P > P-soil-GPS, meaning a complex interaction occurred among glyphosate, phosphate and the soils. PMID- 16312990 TI - Degradation of nitrobenzene in aqueous solution by ozone-ceramic honeycomb. AB - The degradation of nitrobenzene by ceramic-honeycomb catalyzed ozonation was investigated. The results showed that the presence of ceramic honeycombs significantly increased the oxidation rate of nitrobenzene by ozone compared to the case of ozone oxidation alone. In this paper, the effects of various factors on the catalytic oxidation were investigated, such as the amount of catalysts, the ozone dosage, the temperature, the pH value and the presence of tert-butanol. With the addition of tert-butanol the removal of nitrobenzene decreased sharply, which appeared to support that, the degradation of nitrobenzene by ozonation followed a radical type mechanism. The EPR experiments verified that higher nitrobenzene removal rate was attributed to more OH* radicals generated in the catalyzed ozonation than ozonation alone. PMID- 16312991 TI - Distribution of persistent organochlorine pesticides in tissue/organ of silver carp (Hypophthalmichthys molitrix) from Guanting Reservoir, China. AB - The concentration of organochlorine pesticides (OCPs) in tissues and organs of silver carp (Hypophthalmichthys molitrix) from Guanting Reservoir were investigated to evaluate the pollution potential and distribution of OCPs. A total of 16 OCPs were measured and the concentrations were in the range of 1.61 69.01 ng/g wet weight (ww) for total OCPs, 0.16-0.75 ng/g ww for. HCB, 0.75-26.80 ng/g ww for sigmaHCH (sum of alpha-, beta-, gamma- and delta-HCH) and 0.68-35.94 ng/g ww for sigmaDDT( sum of p, p'-DDE, p, p'-DDD, o, p'-DDT and p, p'-DDT). The mean concentrations of total OCPs, HCB, sigmaHCH, and sigmaDDT were 18.04, 0.96, 7.14 and 9.28 ng/g ww, respectively. Among the organochlorine pesticides, beta HCH and p, p'-DDE were the most dominant compounds in tissue and organ with the average concentrations of 4.42 and 8.14 ng/g, respectively. The results obtained in this study show that the levels of 16 OCP residues found in silver carps are low and pose no threat to human health and wildlife fed upon them on the basis of existing related quality guidelines. However, recent input of lindane and DDT might still exist in the area investigated and further investigation should be carried on. PMID- 16312992 TI - Degradation of dyestuff wastewater using visible light in the presence of a novel nano TiO2 catalyst doped with upconversion luminescence agent. AB - A new upconversion luminescence agent, 40CdF2 x 60BaF2 x 0.8Er2O3, was synthesized and its fluorescent spectra were determined. This upconversion luminescence agent can emit five upconversion fluorescent peaks shown in the fluorescent spectra whose wavelengths are all below 387 nm under the excitation of 488 nm visible light. This upconversion luminescence agent was mixed into nano rutile TiO2 powder by ultrasonic and boiling dispersion and the novel doped nano TiO2 photocatalyst utilizing visible light was firstly prepared. The doped TiO2 powder was charactered by XRD and TEM and its photocatalytic activity was tested through the photocatalytic degradation of methyl orange as a model compound under the visible light irradiation emitted by six three basic color lamps. In order to compare the photocatalytic activities, the same experiment was carried out for undoped TiO2 powder. The degradation ratio of methyl orange in the presence of doped nano TiO2 powder reached 32.5% under visible light irradiation at 20 h which was obviously higher than the corresponding 1.64% in the presence of undoped nano TiO2 powder, which indicate the upconversion luminescence agent prepared as dopant can effectively turn visible lights to ultraviolet lights that are absorbed by nano TiO2 particles to produce the electron-cavity pairs. All the results show that the nano rutile TiO2 powder doped with upconversion luminescence agent is a promising photocatalyst using sunlight for treating the industry dye wastewater in great force. PMID- 16312993 TI - Persistent organochlorine residues in sediments of Haihe River and Dagu Drainage River in Tianjin, China. AB - Persistent organochlorine compounds were analyzed by means of GC-ECD in surface sediment samples from two selected rivers in Tianjin, Haihe River and Dagu Drainage River. A total of 16 surface sediment sites were selected along the both rivers. The frequency of detection of T-HCH and T-DDT in sediment samples both was up to 100%, which illustrated that the contamination of HCH and DDT was widespread in Haihe and Dagu Drainage Rivers. Results indicated that the concentrations of various pesticides in sediments from Haihe River were in the range of 3.30-75.96 ng/g dw for T-HCH and 1.57-211.57 ng/g dw for T-DDT. Compared with Haihe River, Dagu Drainage River was contaminated by HCHs and DDTs along the all locations and the values of T-HCH and T-DDT residues in sediments ranged from 2.30 to 124.61 ng/g dw and from 11.28 to 237.30 ng/g dw, respectively. The possible pollution sources were analyzed through monitoring results of organochlorine pesticides(OCPs) residues in sediments from the two rivers. The investigation also indicated that HCH was still used as pesticide in Tianjin partial area. PMID- 16312994 TI - Applying real-time control to enhance the performance of nitrogen removal in CAST system. AB - A bench-scale reactor(72 L) red with domestic sewage, was operated more than 3 months with three operation modes: traditional mode, modified mode and real-time control mode, so as to evaluate effects of the operation mode on the system performance and to develop a feasible control strategy. Results obtained from fixed-time control study indicate that the variations of the pH and oxidation reduction potential(ORP) profiles can represent dynamic characteristics of system and the cycle sequences can be controlled and optimized by the control points on the pH and ORP profiles. A control strategy was, therefore, developed and applied to real-time control mode. Compared with traditional mode, the total nitrogen(TN) removal can be increased by approximately 16% in modified mode and a mean TN removal of 92% was achieved in real-time control mode. Moreover, approximately 12.5% aeration energy was saved in real-time control mode. The result of this study shows that the performance of nitrogen removal was enhanced in modified operation mode. Moreover, the real-time control made it possible to optimize process operation and save aeration energy. PMID- 16312995 TI - Degradation of Populus euphratica community in the lower reaches of the Tarim River, Xinjiang, China. AB - To investigate the relationships between the degradation of plant community and groundwater level in the lower reaches of the Tarim River, nine monitored sections were set along the main stream, where there had been no runoff for nearly 30 years. The characteristics of plant communities were analyzed. It was found that the coverage of trees gradually decreased along the groundwater depth gradient, while the coverage of shrubs slightly increased rather than decreased at first and then gradually decreased, and the coverage of herbs steadily decreased at the beginning and then quickly decreased. The species diversity and species richness of both herbs and woody plants showed obvious degrading trends, while the variations in species evenness were slight. The degrading sequences of species were related to their physiological and ecological characteristics, especially their sensitivity to changes of groundwater table. The herbs with shallow roots first degenerated or disappeared when the groundwater table fell, and then did the deep-rooted herbs, and finally the trees and shrubs with strong tolerance to drought degenerated. The Populus euphratica communities showed typical degrading characteristics, namely the dominant species Populus euphratica remained its dominant status during the degradation. Overall, the existence of strongly tolerant-drought species was the obvious indication of plant species degradation; while simplification of community structure and the decrease of species richness were the obvious indication of plant community degradation. PMID- 16312996 TI - PAHs in aquatic sediment in Hangzhou, China: analytical methods, pollution pattern, risk assessment and sources. AB - Eleven surface sediment samples, from Hangzhou section of Qiantang River and Jinghang Canal, west Lake the inland river were collected to investigate 17 polycyclic aromatic hydrocarbons (PAHs) pollution in aquatic sediments of Hangzhou. Accelerated solvent extraction(ASE) was used to extract PAHs from sediments with satisfactory recoveries. It was found that the total PAHs in the sediments ranged from 308.4 to 3037 ng/g dw, and PAHs pollution in sediments from Jinghang Canal were the heaviest. Lowest effect level (LEL) and severe effect level (SEL) sediment quality guidelines were introduced to perform risk assessment for PAHs pollution in aquatic sediments. Only one sample in Jinghang Canal had adverse impact on benthic organism. 2-3 ring PAHs had a noticeable contribution to total PAHs, especially NA, PHEN. A quantity method was used to determine the major source, the results showed petroleum origin was the chief source to PAHs pollution in all sediments with the exception of sediments from Jinghang Canal where combustion sources had a larger contribution. PMID- 16312997 TI - A novel biosorbent: characterization of the spent mushroom compost and its application for removal of heavy metals. AB - The spent mushroom compost of Lentinus edodes was used as a biosorbent for adsorbing cadmium, lead and chromium from solutions under batch conditions for the first time. Titration of the biomass revealed that it contained at least three types of functional groups. The Fourier transform infrared spectrometry showed that the carboxyl, phosphoryl, phenolic groups were the main groups. The simulated values of pKH and molar quantity were 5.00 and 0.44 mmol/g, 7.32 and 1.38 mmol/g, 10.45 and 1.44 mmol/g, respectively. The biosorption ability increased with pH in acid condition. When 10 mg/L biomass dosage was added in, there was no significant increment of metal uptake. The maximum uptake estimated with the Langmiur isotherm model were 833.33 mg/g for Cd(II), 1000.00 mg/g for Pb(II) and 44.44 mg/g for Cr(III), respectively. All the results showed that vast potential sorption capacity was existed in the biomass for adsorbing these three kinds of metals studied. PMID- 16312998 TI - Involvement of chloride anion in photocatalytic process. AB - The effect of Cl- on photocatalytic degradation (PCD) of pollutants is an important factor since it is ubiquitous in nature. In general, Cl- showed an inhibition on photodegradation due to its scavenging HO* radicals. In this paper, experiments were carried out to examine the effects of Cl- on the PCD of Methylene Blue (MB) and Orange II (OII) in aqueous TiO2 suspensions under UV light illumination. It was found that low concentration of Cl- (< 0.01 mol/L) showed little influence on both dyes, however, high concentration of Cl (> 0.10 mol/L) had a very different influence on the decolorization of dyes: a significant inhibition for MB but a great promotion for OII. In the presence of 0.50 mol/L Cl-, the rate decreased by 70% for MB while increased 7.5-fold for OII. Furthermore, two bands in the ultraviolet region of OII were rapidly broken down. The proposed mechanism was discussed in detail. PMID- 16312999 TI - Effects of adsorption interferents on removal of Reactive Red 195 dye in wastewater by chitosan. AB - Reactive Red 195, which is an azoic anionic dye characterized by the presence of five sulfonic groups and one azoic group, is efficiently removed using chitosan. The increasing chitosan dose had a dramatic positive impact on the achieved color removal, there was approximately a linear relationship between chitosan dose and color removal of dye before color removal reach maximum. Also, the increase of dye concentration led to the increase of chitosan dosage in order to get the same color removal. 92 mg/L of chitosan dosage was sufficient to achieve complete remove of dye at initial concentration of dye at 200 mg/L. For the higher concentrations of dye, high dosages were necessary to reach complete color removal. On the other hand, the use of adsorption interferents (Fe2+, Na+, HCO3- and others) can be interesting, addition of ions had effect on the color removal of Reactive Red 195. Comparing with blank, addition of chemical species approximately decreased the color removal except Na+ and combination of Fe2+ + HCO3-. However, comparing with Fe2+ alone and HCO3- alone, combination of Fe2+ + HCO3- increased the color removal. PMID- 16313000 TI - Survival of pathogenic bacteria in compost with special reference to Escherichia coli. AB - Application of compost in agricultural practice could potentially cause contamination of foodstuffs with pathogenic bacteria such as Escherichia coli O157:H7 (E. Coli O157). We investigated pathogenic bacteria in compost collected from the compost facilities, and evaluated the survival of E. coli K12 and O157 in laboratory experiments. Out of 19 compost product samples, coliform bacteria and salmonella were detected in 7 and 3 samples respectively. The number of coliform bacteria was 1.8 x 10(2) to 2.5 x 10(6) CFU/g dw and that of salmonella was 4.2 x 10(1) to 6.0 x 10(3) CFU/g dw. Moreover, coliform bacteria, fecal coliform, E. coli and salmonella were detected during composting at 54 degrees C to 67 degrees C. The results indicated that moisture content was a very important factor to the heat sensitivity of pathogenic bacteria in compost, E. coil in compost of high moisture content was more sensitive than that in compost of low moisture content, cells harvested in logarithmic phase was more sensitive than these in stationary phase, and E. coli K12 was more sensitive than E. coli O157. Based on the D values, the lethal time of E. coli K12 and O157 from l0(8) to 10(0) CFU/g dw were 16.3 and 28.8 min, respectively, at 60 degrees C in compost with 40% moisture content. However, some E. coil cells survived in composting process at 54 degrees C to 67 degrees C. Water potential (low moisture content) and physiological aspects of bacteria (stationary phase) could explain only in part of the prolonged survival of E. coil in compost, and there should be some other factors that are conducive to bacterial survival in compost. PMID- 16313001 TI - Negative effects of Microcystis blooms on the crustacean plankton in an enclosure experiment in the subtropical China. AB - Effects of Microcystis blooms on the crustacean plankton were studied using enclosure experiments during July-September, 2000. Eight enclosures were set in the hypereutrophic Donghu Lake. Different nutrient concentrations through additional nutrient and sediment in enclosures were expected to result in different abundance of Microcystis. From July to early August, the phytoplankton community was dominated by Chlorophyta, Cryptophyta, Bacillariophyta and Cyanophyta other than Microcystis aeruginosa. M. aeruginosa showed a rapid increase during early August in all enclosures and predominated. Crustacean plankton was dominated by the herbivorous Moina micrura, Diaphanosoma brachyurum and Ceriodaphnia cornuta, and the predaceous Mesocyclops sp. and Thermocyclops taihokuensis. During the pre-bloom period, the dynamics of M. micrura population appeared to be mainly affected by the predaceous cyclopoids. With the development of Microcystis blooms, such interaction between M. micrura and cyclopoids seemed weakened, especially when the Microcystis biomass was high. But there was no apparent influence on the interaction between Leptodora kindti and its zooplanktonic prey. The density of two cyclopoids decreased with the enhancement of Microcystis. The density decline of M. micrura was caused by both predation and inhibition by Microcystis. The low food availability of other edible phytoplankton during the blooms led to low densities of both C. cornuta and D. brachyurum by late August. It appears that dense Microcystis blooms exert strong negative effects on the herbivorous cladocerans and the predaceous cyclopoids. PMID- 16313002 TI - Effects of the surface chemistry of macroreticular adsorbents on the adsorption of 1-naphthol/1-naphthylamine mixtures from water. AB - The adsorption behaviors of 1-naphthol, 1-naphthylamine and 1-naphthol/1 naphthylamine mixtures in water over two macroreticular adsorbents were investigated in single or binary batch systems at 293 K, 303 K and 313 K respectively. All the adsorption isotherms in the studied systems can be adequately fitted by Langmuir model. In the case of aminated macroreticular adsorbent NDA103, 1-naphthol is adsorbed to a larger extent than 1-naphthylamine; while, the opposite trend is found for nonpolar macroreticular adsorbent NDA100. It is noteworthy that at higher temperature (303 K and 313 K), the total uptake amounts of 1-naphthol and 1-naphthylamine in all binary-component systems are obvious larger than the pure uptake amounts in single-component systems, which is presumably due to the cooperative effect primarily arisen from the hydrogen bonding interaction between the loaded 1-naphthol and 1-naphthylamine molecules. The simultaneous adsorption systems were confirmed to be helpful to the selective adsorption towards 1-naphthol according to the larger selective index. PMID- 16313003 TI - Reactions between the SO4*(-) radical and some common anions in atmospheric aqueous droplets. AB - The rate constants of reactions between the SO4*(-) radical and some common anions in atmospheric aqueous droplets e.g. Cl-, NO3-, HSO3- and HCO3- were determined using the laser flash photolysis technique. Absorption spectra of SO4*(-) and the product radicals were also reported. The chloride ion was evaluated among all the anions to be the most efficient scavenger of SO4*(-). The results may supply useful information for a better understanding of the vigorous radical-initiated reactions in atmospheric aqueous droplets such as clouds, rains or fogs. PMID- 16313004 TI - Preliminary studies on methane flux from the ornithogenic soils on Xi-sha atoll, South China Sea. AB - Methane flux from the ornithogenic soils was preliminarily measured by closed chamber method on Xi-sha atoll, South China Sea during March 10 to April 11, 2003 for the first time. The CH4 flux ranged from -4226.7 microg/(m2 x h) to 226.3 microg/(m2 x h) at the observation sites on Dong Island. High atmospheric CH4 consumption was observed from the ornithogenic soils on sunny days. CH4 uptake rates showed the highest value after the midday and they had a strong positive correlation with soil temperatures. Under the same weather conditions, the CH4 fluxes were also observed from the intact and disturbed soils on Yongxing Island. Results showed that the intact soils with natural vegetation also showed high atmospheric CH4 consumption and the average flux was -141.8 microg/(m2 x h). However, disturbed soils via anthropogenic reclamation showed CH4 emissions and the average flux was 441.7 microg/(m2 x h). Therefore land use changes may have an important effect on the CH4 fluxes from the tropical ornithogenic soils. In addition, different observation sites show a high spatial variation in CH4 fluxes. The wetland in salt marsh showed the CH4 emission on Dong Island, and the dry soil sites all showed high atmospheric CH4 consumption, suggesting that CH4 fluxes were predominantly controlled by soil water regime. The effects of soil chemical properties on CH4 fluxes were also analyzed and discussed in this paper. PMID- 16313005 TI - Achieving and maintaining biological nitrogen removal via nitrite under normal conditions. AB - The principal aim of this paper is to develop an approach to realize stable biological nitrogen removal via nitrite under normal conditions. Validation of the new method was established on laboratory-scale experiments applying the sequencing batch reactor (SBR) activated sludge process to domestic wastewater with low C/N ratio. The addition of sodium chloride (NaCl) to influent was established to achieve nitrite build-up. The high nitrite accumulation, depending on the salinity in influent and the application duration of salt, was obtained in SBRs treating saline wastewater. The maintenance results indicated that the real time SBRs can maintain stable nitrite accumulation, but conversion from shorter nitrification-denitrification to full nitrification-denitrification was observed after some operation cycles in the other SBR with fixed-time control. The presented method is valuable to offer a solution to realize and to maintain nitrogen removal via nitrite under normal conditions. PMID- 16313006 TI - A cleaner two-step synthesis of high purity diallyldimethylammonium chloride monomers for flocculant preparation. AB - In order to improve the flocculation efficiency of polydiallyldimethylammonium chloride (PDADMAC), high molecular weight PDADMAC should be prepared from high purity diallyldimethylammonium chloride(DADMAC) monomers. In this paper, a cleaner method with microwave irradiation and alkali solidification was proposed for preparing high pure DADMAC by selective heating under low temperature, and the prepared high purity DADMAC is characterized using FTIR and atomic absorption spectrometry. The new method provides a solution to the key technical problem of PDADMAC synthesis. Comparing with the conventional methods, the results showed that the advantages of the novel synthesis include: (a) high purity DADMAC is improved from 57% to 71%; (b) reaction time of tertiary amine preparation is shortened from 6 h to 7 min; (c) water instead of acetone was used as reaction medium; (d) toxic by-products, wastewater and waste gas are eliminated. Flocculant made from the synthesized high purity DADMAC monomers was proved more efficient in flocculation tests. PMID- 16313007 TI - Changes in enzymes activity, substrate utilization pattern and diversity of soil microbial communities under cadmium pollution. AB - Heavy metal pollution has received increasing attention in recent years mainly because of the public awareness of environmental issues. In this study we have evaluated the effect of cadmium (Cd) on enzymes activity, substrate utilization pattern and diversity of microbial communities in soil spiked with 0, 20, 40, 60, 80, and 100 mg/kg Cd, during 60 d of incubation at 25 degrees C. Enzyme activities determined at 0, 15, 30, 45, and 60 d after heavy metal application (DAA) showed marked declines for various Cd treatments, and up to 60 DAA, 100 mg/kg Cd resulted in 50.1%, 47.4%, and 39.8% decreases in soil urease, acid phosphatase and dehydrogenase activities, respectively to control. At 60 DAA, substrate utilization pattern of soil microbial communities determined by inoculating Biolog ECO plates indicated that Cd addition had markedly inhibited the functional activity of soil microbial communities and multivariate analysis of sole carbon source utilization showed significantly different utilization patterns for 80 and 100 mg/kg Cd treatments. The structural diversity of soil microbial communities assessed by PCR-DGGE method at 60 DAA, illustrated that DGGE patterns in soil simplified with increasing Cd concentration, and clustering of DGGE profiles for various Cd treatments revealed that they had more than 50% difference with that of control. PMID- 16313008 TI - Numerical simulation of a combined oxidation ditch flow using 3D k-epsilon turbulence model. AB - The standard three dimensional(3D) k-epsilon turbulence model was applied to simulate the flow field of a small scale combined oxidation ditch. The moving mesh approach was used to model the rotor of the ditch. Comparison of the computed and the measured data is acceptable. A vertical reverse flow zone in the ditch was found, and it played a very important role in the ditch flow behavior. The flow pattern in the ditch is discussed in detail, and approaches are suggested to improve the hydrodynamic performance in the ditch. PMID- 16313009 TI - Heavy metal accumulation of urban domestic rubbish compost in turfgrass by EDTA chelating. AB - Seven kinds of heavy metal accumulation of rubbish compost in turfgrass by EDTA chelating were investigated. With EDTA application, heavy metal accumulation by two species of turfgrass was increased significantly. The enrichment coefficients of Lolium perenne L(L) and Festuca arundinacea L(F) to Cr reached 9.45 and 6.15 respectively. In the range of EDTA dosages given, heavy metal accumulation in turfgrass increased with increasing EDTA level. There were significant differences in remediation of different metals by applying EDTA. L had high ability to accumulate Cr, Cd, Ni and Zn, showing better remediation to heavy metals of rubbish compost. In contrast, F showed high ability to accumulate Cr, Ni, Cu and Zn. Low EDTA level increased aboveground net primary production (ANP) of turfgrass, but EDTA would considerably inhibit it when EDTA was higher than 20 mmol/kg. The results demonstrated that the optimum dosage of EDTA for remediating heavy metals in rubbish compost by turfgrass was between 10 mmol/kg and 20 mmol/kg. PMID- 16313010 TI - Uptake and accumulation of phenanthrene and pyrene in spiked soils by Ryegrass (Lolium perenne L.). AB - Phytoremediation has long been recognized as a cost-effective method for the removal of polycyclic aromatic hydrocarbons (PAHs) from soil. A study was conducted to investigate the uptake and accumulation of PAHs in root and shoot of Lolium perenne L. Pot experiments were conducted with series of concentrations of 3.31-378.37 mg/kg for phenanthrene and those of 4.22-365.38 mg/kg for pyrene in a greenhouse. The results showed that both ryegrass roots and shoots did take up PAHs from spiked soils, and generally increased with increasing concentrations of PAH in soil. Bioconcentration factors (BCFs) of phenanthrene by shoots and roots were 0.24--4.25 and 0.17-2.12 for the same treatment. BCFs of pyrene by shoots were 0.20--1.5, except for 4.06 in 4.32 mg/kg treatment, much lower than BCFs of pyrene by roots (0.58--2.28). BCFs of phenanthrene and pyrene tended to decrease with increasing concentrations of phenanthrene and pyrene in soil. Direct uptake and accumulation of these compounds by Lolium perenne L. was very low compared with the other loss pathways, which meant that plant-promoted microbial biodegradation might be the main contribution to plant-enhanced removal of phenanthrene and pyrene in soil. However, the presence of Lolium perenne L. significantly enhanced the removal of phenanthrene and pyrene in spiked soil. At the end of 60 d experiment, the extractable concentrations of phenanthrene and pyrene were lower in planted soil than in non-planted soil, about 83.24%--91.98% of phenanthrene and 68.53%-84.10% of pyrene were removed from soils, respectively. The results indicated that the removal of PAHs in contaminated soils was a feasible approach by using Lolium perenne L. PMID- 16313011 TI - Oxidative stress related enzymes in response to chromium (VI) toxicity in Oxya chinensis (Orthoptera: Acridoidae). AB - The toxic effects of Cr(VI) on antioxidant enzymes of Oxya chinensis (Orthoptera: Acridoidae) were determined. Changes in the activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and guaiacol peroxidase (GPx) were measured in O. chinensis insects injected with Cr(VI). Fifth-nymphs of O. chinensis insects were injected with Cr(VI) with different concentrations (0, 75, 150, 225, 300, 375, 450 mg/kg of body weight). The results showed that Cr(VI) led to the change of SOD, CAT, and GPx activities at different concentrations, which revealed that: (1) The oxidative stress of SOD increased with the increase of Cr(VI) concentration. (2) With the increase of Cr(VI) concentrations, CAT activities for females increased at lower concentrations, but decreased at higher concentration range, which indicated that antioxidant system of O. chinensis was not influenced by the presence of Cr(VI). A very similar response to Cr(VI) effect for males indicated that Cr(VI) concentrations were not high enough to damage O. chinensis in terms of CAT. (3) The GPx activity for females increased in all treatments, which revealed that the damage power of Cr(VI) was increased with the increase of Cr(VI) concentrations in terms of GPx, but the effect was not so remarkable. There was not a consistent trend of GPx activities for males in all treatments of Cr(VI). Cr (VI)-induced changes in antioxidant enzymes were different for SOD, CAT and GPx, of which the tendency was that activities generally changed with increase of concentrations of Cr(VI) suggesting SOD, CAT, and GPx could serve as indices of oxidative stress to some extent. PMID- 16313012 TI - A pilot-scale jet bubbling reactor for wet flue gas desulfurization with pyrolusite. AB - MnO2 in pyrolusite can react with SO2 in flue gas and obtain by-product MnSO4 x H2O. A pilot scale jet bubbling reactor was applied in this work. Different factors affecting both SO2 absorption efficiency and Mn2+ extraction rate have been investigated, these factors include temperature of inlet gas flue, ration of liquid/solid mass flow rate (L/S), pyrolusite grade, and SO2 concentration in the inlet flue gas. In the meantime, the procedure of purification of absorption liquid was also discussed. Experiment results indicated that the increase of temperature from 30 to 70 K caused the increase of SO2 absorption efficiency from 81.4% to 91.2%. And when SO2 concentration in the inlet flue gas increased from 500 to 3000 ppm, SO2 absorption efficiency and Mn2+ extraction rate decreased from 98.1% to 82.2% and from 82.8% to 61.7%, respectively. The content of MnO2 in pyrolusite had a neglectable effect on SO2 absorption efficiency. Low L/S was good for both removal of SO2 and Mn2+ extraction. The absorption liquid was filtrated and purified to remove Si, Mg, Ca, Fe, Al and heavy metals, last product MnSO4 x H2O was obtained which quality could reach China GB1622-86, the industry grade standards. PMID- 16313013 TI - Effect of heavy metals on soil microbial activity and diversity in a reclaimed mining wasteland of red soil area. AB - The microbial biomass, basal respiration and substrate utilization pattern in copper mining wasteland of red soil area, southern China, were investigated. The results indicated that soil microflora were obviously different compared with that of the non-mine soil. Microbial biomass and basal respiration were negatively affected by the elevated heavy metal levels. Two important microbial ecophysiological parameters, namely, the ratio of microbial biomass C(Cmic)/organic C(Corg) and metabolic quotient(qCO2) were closely correlated to heavy metal stress. There was a significant decrease in the Cmic/Corg ratio and an increase in the metabolic quotient with increasing metal concentration. Multivariate analysis of Biolog data for sole carbon source utilization pattern demonstrated that heavy metal pollution had a significant impact on microbial community structure and functional diversity. All the results showed that soil microbiological parameters had great potential to become the early sensitive, effective and liable indicators of the stresses or perturbations in soils of mining ecosystems. PMID- 16313014 TI - A new design of groundwater sampling device and its application. AB - Compounds in the atmosphere contaminate samples of groundwater. An inexpensive and simple method for collecting groundwater samples is developed to prevent contamination when the background concentration of contaminants is high. This new design of groundwater sampling device involves a glass sampling bottle with a Teflon-lined valve at each end. A cleaned and dried sampling bottle was connected to a low flow-rate peristaltic pump with Teflon tubing and was filled with water. No headspace volume was remained in the sampling bottle. The sample bottle was then packed in a PVC bag to prevent the target component from infiltrating into the water sample through the valves. In this study, groundwater was sampled at six wells using both the conventional method and the improved method. The analysis of trichlorofluoromethane (CFC-11) concentrations at these six wells indicates that all the groundwater samples obtained by the conventional sampling method were contaminated by CFC-11 from the atmosphere. The improved sampling method greatly eliminated the problems of contamination, preservation and quantitative analysis of natural water. PMID- 16313015 TI - Reuse rate of treated wastewater in water reuse system. AB - A water quality model for water reuse was made by mathematics induction. The relationship among the reuse rate of treated wastewater (R), pollutant concentration of reused water (Cs), pollutant concentration of influent (C0), removal efficiency of pollutant in wastewater (E), and the standard of reuse water were discussed in this study. According to the experiment result of a toilet wastewater treatment and reuse with membrane bioreactors, R would be set at less than 40%, on which all the concemed parameters could meet with the reuse water standards. To raise R of reuse water in the toilet, an important way was to improve color removal of the wastewater. PMID- 16313016 TI - Synergistic removal of nitrogen monoxide by non-thermal plasma and catalyst simultaneously. AB - An experimental system of De-NO with plasma-catalyst(Cu zeolite) was established to investigate the differences between De-NO with plasma-catalyst and De-NO only with plasma, to provide the instruction for selecting appropriate catalyst and operating condition. The characteristics of De-NO with plasma and De-NO with plasma-catalyst were investigated comparatively by experiments. The experimental results show that De-NO with plasma-catalyst has high NO removal rate; Cu zeolite is an effective catalyst which can promote NO removal rate in plasma remarkably; De-NO with plasma-catalyst should be operated at low temperature and the temperature has opposite effects on the function of catalyst and plasma; water vapor and O2 can increase the NO removal rate. PMID- 16313017 TI - Enhanced catalytic degradation process of o-nitrochlorobenzene by palladium catalyzed fe0 particles. AB - Over Pd/Fe bimetallic catalyst, o-nitrochlorobenzene (o-NCB), at a concentration of 20 mg/L in aqueous solutions, is rapidly converted to o-chloroaniline (o-CAN) first, and then quickly dechlorinated to aniline(AN) and Cl-, without other intermediate reaction products. The aminated and dechlorinated reactions are believed to take place on the surface site of the Pd/Fe. The o-NCB removal efficiency and the next dechlorination rate increase with an increase of bulk loading of palladium and catalysts addition due to the increase of both the surface loading of palladium and the total surface area. These results indicate that reduction, amination and dechlorination of o-NCB by palladium-catalyzed Fe0 particles, can be designed for remediation of contaminated groundwater. PMID- 16313018 TI - Qualitative analysis and quantitative simulation on Yin-Huang water salinization mechanism in Bei-Da-Gang Reservoir. AB - Yellow River water transfer for Tianjin is important in solving the water shortage in Tianjin, which facilitate economic development and social progress for many years. Fresh water drawn from Yellow River (i.e., Yin-Huang water) becomes saltier and saltier when being stored in the Bei-Da-Gang reservoir. We qualitatively analyze the water salinization mechanism based on mass transfer theory. The main factors are salinity transfer of saline soil, evaporation concentrating, and the agitation of wind. A simulative experimental pond and an evaporation pond were built beside the Bei-Da-Gang reservoir to quantitatively investigate the water salinization based on water and solute balance in the simulative pond. 80% of increased [Cl-] is due to the salinity transfer of the saline soil and the other 20% is due to evaporation concentrating, so the former is the most important factor. We found that the salinization of Yin-Huang water can be described with a zero-dimension linear model. PMID- 16313019 TI - Environmental characteristics of sandstorm of Minqin Oasis in China for recent 50 years. AB - The purpose of this present study is to investigate the frequency and variation of sandstorm in Minqin Oasis. Using daily observational data of sandstorm and other meteorologic data from 1954 to 2000, we have illuminated the relationship between sandstorm, meteorological parameters and human activities. The results of the analysis show that the highest frequency of sandstorms occurrence and their duration mainly focus on March, April and May, especially in April. Most of sandstorms occur from midday to nightfall, but relative few appear from midnight to forenoon, which apparently correlates to the daily variations of atmospheric thermal stability within atmospheric boundary layer. Monthly mean and annual mean duration of sandstorms coincide well with the frequencies of sandstorm occurrence. PMID- 16313020 TI - A model supported by GIS for locating and quantifying PM2.5 emission originated from residential wood burning. AB - A research method was presented for spatially quantifying and allocating the potential activity of a fine particle matter emission (PM2.5), which originated from residential wood burning (RWB) in this study. Demographic, hypsographic, climatic and topographic data were compiled and processed within a geographic information system (GIS), and as independent variables put into a linear regression model for describing spatial distribution of the potential activity of residential wood burning as primary heating source. In order to improve the estimation, the classifications of urban, suburban and rural were redefined to meet the specifications of this application. Also, several definitions of forest accessibility were tested for estimation. The results suggested that the potential activity of RWB was mostly determined by elevation of a location, forest accessibility, urban/non-urban position, climatic conditions and several demographic variables. The linear regression model could explain approximately 86% of the variation of surveyed potential activity of RWB. The analysis results were validated by employing survey data collected mainly from a WebGIS based phone interview over the study area in central California. Based on lots free public GIS data, the model provided an easy and ideal tool for geographic researchers, environmental planners and administrators to understand where and how much PM2.5 emission from RWB was contributed to air quality. With this knowledge they could identify regions of concern, and better plan mitigation strategies to improve air quality. Furthermore, it allows for future adjustment on some parameters as the spatial analysis method is implemented in the different regions or various eco-social models. PMID- 16313021 TI - No difference in activity of Sod-1 genotypes of Poa annua to short-term treatment of ambient gaseous organic pollution. AB - Our previous studies indicated that genotypes at locus Sod-1 of Poa annua changed clinally along a gradient of gaseous organic pollution. In the present study, we aimed to know whether there were differential responses of activities of different superoxide dismutase (SOD) genotypes to short-term treatment of ambient gaseous organic pollution. Significant bias from Hardy-Weinberg equibrium was observed on locus Sod-1, and no genotype Sod-1-BB was found. Significantly increased activities were observed in most treatments for genotype Sod-1-AA and in one treatment for genotype Sod-1-AB. However, no significant difference in SOD activities was found between the two genotypes. It was interpreted that fitness difference between the two genotypes was small and treatment duration was too short or extent of ambient organic pollution was too low to lead to differential responses. Other environmental factor effects on activities of superoxide dismutases can also explain the results. PMID- 16313022 TI - Photocatalytic degradation of Chicago Sky Blue 6B and Benzopurpurin 4B using titanium dioxide thin film. AB - Aqueous solutions of azo dyes undergo degradation to form harmless intermediates and colorless products following irradiation by visible light in the presence of titanium dioxide thin films. The dyes that were studied in this work are: Chicago Sky Blue 6B and Benzopurpurin 4B. Results obtained indicated that complete mineralization of the dyes took place under the experimental conditions. There was an increase in conductivity after the complete mineralization experiments possibly indicating the formation of ions such as NO3- and SO4(2-). Chemical oxygen demand (COD) measurements show a decrease in organic matter for both dyes following complete degradation. The effect of how changing experimental conditions such as pH, temperature and starting concentrations of dyes affected the rate of dye degradation was measured. There was an increase in the rate of disappearance of the dye color at lower pH. High concentrations of dye solutions required long degradation time. PMID- 16313023 TI - Simultaneous anaerobic-aerobic biodegradation of halogenated phenolic compound under oxygen-limited conditions. AB - The successful application of co-immobilized aerobic-anaerobic biomass under limited aeration in wastewater treatment systems would eliminate the problems associated with the intermediates mono-chlorophenol (MCP) and di chlorophenol(DCP) accumulations. With low initial pentachlorophenol (PCP) concentration, all PCP could be completely removed under oxygen-limited strict anaerobic conditions, and the removal efficiencies with different initial headspace oxygen percentage (IHOP) were not obviously different from each other. While at high initial PCP concentration, under strictly anaerobic conditions PCP and their intermediates were clearly higher than that under other conditions, and produced obvious accumulation, the highest PCP reduction was achieved by the system receiving 30 IHOP, oxygen-limited system also exhibited lower residual TOC concentration and lower concentration of metabolic intermediates MCP and DCP. These results suggested that under strictly anaerobic condition the reductive dechlorination of low chlorinated compounds became rate limiting in the reductive dechlorination pathway, less chlorinated compounds be more amenable to aerobic degradation, and the aerobes of outer layers could function under limited oxygen. The co-immobilized aerobic-anaerobic biomass for methanogenesis under limited aeration for chlorophenol degradation might be an attractive and efficient alternative for the sequential anaerobic/aerobic system to achieve mineralization of a broad range of recalcitrance highly chlorinated organics and low final TOC concentrations. PMID- 16313024 TI - Purified terephthalic acid wastewater biodegradation and toxicity. AB - The biodegradation and toxicity of the purified terephthalic acid (PTA) processing wastewater was researched at NJYZ pilot with the fusant strain Fhhh in the carrier activated sludge process (CASP). Sludge loading rate (SLR) for Fhhh to COD of the wastewater was 1.09 d(-1) and to PTA in the wastewater was 0.29 d( 1). The results of bioassay at the pilot and calculation with software Ebis3 showed that the 48h-LC50 (median lethal concentration) to Daphnia magna for the PTA concentration in the wastewater was only 1/10 of that for the chemical PTA. There were 5 kinds of benzoate pollutants and their toxicities existing in the wastewater at least. The toxicity parameter value of the pure chemical PTA cannot be used to predicate the PTA wastewater toxicity. The toxicity of the NJYZ PTA wastewater will be discussed in detail in this paper. PMID- 16313025 TI - Federal veterinary team deployed post-Rita raises questions. PMID- 16313026 TI - AVMA GHLIT responds to health care fraud. PMID- 16313027 TI - Standards taking shape for health, welfare of fish. PMID- 16313028 TI - Animals, people still recovering from hurricane. PMID- 16313029 TI - Finds interesting slant on rising educational debt. PMID- 16313030 TI - The case for canine-exclusive practice. PMID- 16313031 TI - What is your diagnosis? Benign bone cysts. PMID- 16313032 TI - Theriogenology question of the month. Cystic remnants of the mesonephric (wolffian) ducts (ie, Gartner cyst). PMID- 16313033 TI - Survey of wildlife rehabilitators regarding rabies vector species. AB - OBJECTIVE: To evaluate the risks associated with wildlife rehabilitation and the reemergence of wildlife rabies in North Carolina through assessment of the status of knowledge and attitudes of licensed in-state wildlife rehabilitators about rabies and rabies vector species (RVS). DESIGN: Questionnaire survey. SAMPLE POPULATION: 672 North Carolina licensed wildlife rehabilitators registered in 1999. PROCEDURE: Wildlife rehabilitators were contacted by mail to determine their status of knowledge and attitudes regarding rabies and RVS. The questionnaire was designed to determine rehabilitators' recent experiences with RVS, attitudes toward regulations, and knowledge of rabies virus transmission. Results were analyzed by use of the chi2 test. RESULTS: Questionnaire responses were provided by 210 of the 672 (31.3%) wildlife rehabilitators. Among rehabilitators, there were some inconsistencies in their knowledge base regarding rabies (eg, 25% reported that they did not know at what age animals were capable of transmitting rabies virus). Most respondents were amenable to all proposed licensing prerequisites for handling RVS (ie, record keeping, additional training, and veterinarian support). Respondents reported > 580 calls annually about rehabilitating RVS, and 80% believed at least some of their peers were rehabilitating RVS illegally. CONCLUSIONS AND CLINICAL RELEVANCE: With the establishment of rabies as a disease that is endemic among wildlife species in North Carolina, educational efforts directed at wildlife rehabilitators (a subpopulation of residents potentially at high risk of rabies virus infection) would have direct and indirect public health benefits; similar efforts may be useful to public health communities elsewhere in the United States. PMID- 16313034 TI - Effect of formulation and route of administration on tissue residues and withdrawal times. PMID- 16313035 TI - A comprehensive review of housing for pregnant sows. PMID- 16313036 TI - National survey of owner-directed aggression in English Springer Spaniels. AB - OBJECTIVE: To determine prevalence of owner-directed aggression and identify associated environmental and genetic factors in English Springer Spaniels. DESIGN: Prevalence survey. ANIMALS: 1,053 adult English Springer Spaniels. PROCEDURE: A mail survey was sent to 2,400 randomly selected owners of adult American Kennel Club-registered English Springer Spaniels. Dogs with a history of aggression to family members and familiar humans were compared with dogs without such a history. RESULTS: 1,053 questionnaires (56.1% of the 1,877 delivered) were completed. A history of owner-directed growling or more intense aggression was reported in 510 (48.4%) dogs. Two hundred seventy-seven (26.3%) dogs had bitten a human in the past; 65.2% of bites were directed at familiar (owner or nonowner) adults and children. Variables associated with owner-directed aggression included sex of dog (male), neuter status (neutered, regardless of sex), show or bench lines, age > 4 years, aggression to unfamiliar adults and children, acquisition from a hobby breeder, less responsiveness to obedience cues, and a specific kennel and 1 popular sire from that kennel in a 4-generation pedigree. CONCLUSIONS AND CLINICAL RELEVANCE: Owner-directed aggression in adult English Springer Spaniels was associated with a number of environmental, sex-related, and inherited factors. To reduce the risk of aggression, prospective owners might seek a female, hunting-type English Springer Spaniel from an experienced breeder. However, because risk factors are broad and varied, there are limitations to the extent to which behavior can be predicted and further study is needed of the inheritance of aggression in this breed. PMID- 16313037 TI - Estimate of the annual economic impact of treatment of cranial cruciate ligament injury in dogs in the United States. AB - OBJECTIVE: To estimate the economic impact to veterinary clients for the medical and surgical treatment of rupture of the cranial cruciate ligament (RCCL) in dogs for the year 2003. DESIGN: Economic impact survey. SAMPLE POPULATION: 501 diplomates of the American College of Veterinary Surgeons (ACVS) indicating that their area of surgical emphasis was small animal orthopedic surgery or small animal general and orthopedic surgery and 4,000 veterinarians indicating to the AVMA that their professional area was small animal practice exclusive or mixed animal practice (at least 80% small animal). PROCEDURE: Veterinarians were surveyed concerning the cost for medical and surgical treatment of RCCL for 2003. The economic impact was calculated by multiplying the number of RCCL surgeries performed by the mean cost of surgery. This was added to the number of RCCL cases managed medically multiplied by the mean cost of medical management. This estimate for survey responders was extrapolated to the total number of veterinarians in the study population for the ACVS or AVMA. RESULTS: Estimates for the total cost of surgery were $171,730,134.72 and $1,020,167,907 for veterinarians in the ACVS and AVMA populations, respectively. The cost of medical management was $2,885,687.86 and $126,558,155.16 for veterinarians in the ACVS and AVMA populations, respectively. After combining the ACVS and AVMA populations, we estimated that owners spent $1.32 billion for the treatment of RCCL in the United States in 2003. CONCLUSIONS AND CLINICAL RELEVANCE: RCCL is a prevalent, costly injury. Results may motivate veterinary and consumer agencies to prioritize funding for a better understanding of the injury. PMID- 16313038 TI - Myokymia and neuromyotonia in a cat. AB - A 6-year-old spayed female domestic shorthair cat was examined because of a 2 week history of rhythmic muscle movements. Physical examination revealed thoracic limb rigidity, contracture of the carpi, generalized muscle atrophy, and rhythmic rippling of the muscles of all 4 limbs. Results of a CBC and serum biochemistry profile were unremarkable other than high creatine kinase activity. Electromyography revealed unique high-frequency discharges, including rhythmic bursts of single motor unit potentials appearing as doublets (myokymia) and more prolonged bursts of nonrhythmic motor unit potentials with characteristic waning amplitudes (neuromyotonia). Histologic examination of muscle biopsy specimens revealed noninflammatory necrotizing myopathy with regeneration. The cat did not respond to treatment with carbamazepine or prednisone but improved rapidly after treatment with phenytoin was initiated. Six months after initial examination, electromyography revealed a substantial decrease in the amount of spontaneous activity in previously affected muscles. However, the myokymic and neuromyotonic discharges were still present, albeit with a substantial decrease in frequency. PMID- 16313039 TI - Sarcoma of the proximal portion of the tibia in a dog 5.5 years after tibial plateau leveling osteotomy. AB - Osseous neoplasia was identified in the proximal portion of the tibia and distal portion of the femur in an 11.75-year-old spayed female German Shepherd Dog. A tibial plateau leveling osteotomy, followed by application of a metal plate, had been performed on the affected limb 5.5 years earlier. Areas of osteolysis and periosteal proliferation were seen radiographically, with an intense area of osteolysis directly beneath the metal plate. Histologically, the tumor was identified as a poorly differentiated sarcoma. Extracellular and intracellular debris was seen histologically, and energy-dispersive x-ray analysis confirmed that this debris was metallic. On visual examination, areas of the underside of the metal plate that had been in contact with bone had a dull, roughened appearance, and scanning electron microscopy of these areas revealed multiple corrosion pits. The plate was strongly magnetic, suggesting that it contained ferrite, and metallographic examination of the plate revealed substantial differences in the chemical makeup of various parts of the plate. Microstructure analysis revealed that the plate consisted of an austenite matrix with a large fraction of ferrite. The plate was determined to be a cast 316L stainless steel implant, but it did not meet American Society for Testing Materials standards for implant-grade materials. The possibility that implant corrosion might have played a role in tumor development is of concern; however, a definitive association was not established. PMID- 16313040 TI - Correlation between hepatobiliary scintigraphy and surgery or postmortem examination findings in dogs and cats with extrahepatic biliary obstruction, partial obstruction, or patency of the biliary system: 18 cases (1995-2004). AB - OBJECTIVE: To evaluate the usefulness of serum biochemical variables and scintigraphic study results for differentiating between dogs and cats with complete extrahepatic biliary obstruction (EHO) and those with partial EHO or patent bile ducts. STUDY DESIGN: Retrospective case series. ANIMALS: 17 dogs and 1 cat. PROCEDURE: Animals that underwent hepatobiliary scintigraphy and had either surgical or postmortem confirmation of the degree of bile duct patency were included. Scintigraphic images were evaluated and biliary tracts were classified as patent, partially obstructed but patent, or obstructed. Surgery or postmortem examination was considered the gold standard for diagnosis, and compared with those findings, sensitivity and specificity of scintigraphy were calculated. RESULTS: With absence of radioactivity in the intestinal tract as the diagnostic criterion for EHO, the sensitivity and specificity of scintigraphic diagnosis were both 83% when final images were acquired at 19 to 24 hours, compared with 100% and 33%, respectively, when 180 minutes was used as the cutoff time. Animals with partial biliary obstruction had less intestinal radioactivity that arrived later than that observed in animals with patent biliary tracts. CONCLUSIONS AND CLINICAL RELEVANCE: Animals in which intestinal radioactivity has not been observed after the standard 3 to 4 hours should undergo additional scintigraphic imaging. Findings in animals with partial biliary obstruction include delayed arrival of radioactivity and less radioactivity in the intestine. Distinguishing between complete and partial biliary tract obstruction is important because animals with partial obstruction may respond favorably to medical management and should not be given an erroneous diagnosis of complete obstruction. PMID- 16313041 TI - Prognostic factors associated with survival two years after surgery in dogs with malignant mammary tumors: 79 cases (1998-2002). AB - OBJECTIVE: To identify prognostic factors for female dogs that have undergone surgical removal of malignant mammary tumors. DESIGN: Retrospective case series. ANIMALS: 79 female dogs with malignant mammary tumors. PROCEDURE: Information obtained from the medical records included breed, age, sex, tumor size (maximum diameter), number and location of affected mammary glands, time between tumor identification and surgical removal, radiographic evidence of distant metastasis, surgical procedure, ovariohysterectomy (OHE) status, histologic classification of the tumor, and survival time. RESULTS: Results of univariate analyses indicated that clinical stage, tumor size, OHE status, metastasis to adjacent lymph nodes or distant sites, and histologic classification of the tumor were significantly associated with survival 2 years after surgery. Tumors > or = 5 cm in diameter and tumors that had been identified > 6 months before surgery were more likely to metastasize to adjacent lymph nodes. Ovariohysterectomy was more beneficial in dogs with complex carcinomas than in dogs with simple carcinomas. In multivariate analyses, clinical stage, tumor size, and OHE status were significantly associated with survival 2 years after surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that tumor stage, tumor size, and OHE status were significant prognostic factors associated with survival 2 years after surgery in dogs with malignant mammary tumors. Further, either dogs with tumors > or = 5 cm in diameter or dogs with tumors present for > 6 months prior to surgery had a higher risk of having lymph node metastases. PMID- 16313042 TI - Use of color Doppler echocardiography to assess the development of valvular regurgitation in Standardbred trotters. AB - OBJECTIVE: To evaluate the incidence and development of valvular regurgitation and its effect on racing performance in Standardbred trotters from 2 to 3.5 years of age. DESIGN: Longitudinal observational study. ANIMALS: 103 horses. PROCEDURE: Horses were examined via echocardiographic imaging 4 times at 6-month intervals. Two-dimensional and M-mode echocardiographic examinations were performed in each horse, and color Doppler imaging was used to detect and evaluate regurgitant blood flow at each valve. RESULTS: For all cardiac valves, the frequency of valvular regurgitation increased significantly during the 18-month study period. Regurgitation was mild in all valves in which it was detected, and no progression in severity was observed at any valve. Tricuspid regurgitation was more prevalent in horses that raced regularly and was diagnosed more frequently in males and horses with larger left ventricular mass and left ventricular internal diameter during diastole. No relationship between racing performance and valvular regurgitation was observed. CONCLUSIONS AND CLINICAL RELEVANCE: The incidence of valvular regurgitation increased during the study period, and it was common at 3.5 years of age. It was not possible to determine whether the increase was a result of maturity or training. The study revealed that there is a high prevalence of mild valvular regurgitation at this age, but apparently, the regurgitation had no effect on racing performance in this age group. Whether regurgitation will progress or influence future performance is not known. PMID- 16313043 TI - Effects of treatment with omeprazole or ranitidine on gastric squamous ulceration in racing Thoroughbreds. AB - OBJECTIVE: To compare the effects of oral administration of omeprazole and ranitidine on gastric squamous ulceration in Thoroughbreds in race training. DESIGN: Modified crossover study. ANIMALS: 60 Thoroughbreds in race training with gastric squamous mucosal ulceration. PROCEDURE: Horses were randomly allocated into 3 groups. Group 1 received no treatment for 28 days followed by administration of omeprazole (4 mg/kg [1.8 mg/lb], PO, once daily) for 28 days; group 2 received omeprazole (4 mg/kg, PO, once daily) for 28 days followed by no treatment for 28 days; and group 3 received ranitidine (6.6 mg/kg [3.0 mg/lb], PO, q 8 h) for 28 days followed by administration of omeprazole (4 mg/kg, PO, once daily) for 28 days. Ulceration was assessed endoscopically at days 0, 28, 42, and 56. Lesions were scored from 0 (no ulceration) to 3 (severe ulceration). RESULTS: After the initial 28 days of treatment, the decrease in ulcer severity was significantly greater after omeprazole treatment than after ranitidine treatment. Ulcer severity decreased significantly in group 3 horses after 14 days of treatment with omeprazole. Discontinuation of omeprazole resulted in worsening of ulcer scores; however, ulcer scores at completion of the study were less than at day 0. Horses that received omeprazole after 28 days of ranitidine treatment had a further reduction in ulcer severity. CONCLUSIONS AND CLINICAL RELEVANCE: Omeprazole was more effective than ranitidine in healing gastric squamous ulcers in Thoroughbreds in race training. Improvement was detected by 14 days and persisted in most of the group 2 horses for at least 28 days after omeprazole treatment was discontinued. PMID- 16313044 TI - Evaluation of five commercially available assays and measurement of serum total protein concentration via refractometry for the diagnosis of failure of passive transfer of immunity in foals. AB - OBJECTIVE: To determine and compare sensitivity, specificity, accuracy, and predictive values of measurement of serum total protein concentration by refractometry as well as 5 commercially available kits for the diagnosis of failure of passive transfer (FPT) of immunity in foals. DESIGN: Prospective study. ANIMALS: 65 foals with various medical problems and 35 clinically normal foals. PROCEDURE: IgG concentration in serum was assessed by use of zinc sulfate turbidity (assay C), glutaraldehyde coagulation (assay D), 2 semiquantitative immunoassays (assays F and G), and a quantitative immunoassay (assay H). Serum total protein concentration was assessed by refractometry. Radial immunodiffusion (assays A and B) was used as the reference method. RESULTS: For detection of IgG < 400 mg/dL, sensitivity of assay H (100%) was not significantly different from that of assays C, E, and G (88.9%). Specificity of assays H (96.0%) and G (95.8%) was significantly higher than that of assays C (79.4%) and E (78.1 %). For detection of IgG < 800 mg/dL, sensitivities of assays H (976%), D (92.9%), C (81.0%), and G (81.0%) were significantly higher than that of assay F (52.4%). Specificity of assays F (100%), G (94.7%), and H (82.8%) was significantly higher than that of assays C (56.9%) and D (58.6%). Serum total protein concentration < or = 4.5 g/dL was suggestive of FPT, whereas values > or = 6.0 g/dL indicated adequate IgG concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Most assays were adequate as initial screening tests. However, their use as a definitive test would result in unnecessary treatment of foals with adequate IgG concentrations. PMID- 16313045 TI - Musculoskeletal problems associated with lameness and poor performance among horses used for barrel racing: 118 cases (2000-2003). AB - OBJECTIVE: To identify types of musculoskeletal problems associated with lameness or poor performance in horses used for barrel racing. DESIGN: Retrospective case series. ANIMALS: 118 horses. PROCEDURE: Medical records were reviewed for information on signalment, history, physical and lameness examination findings, diagnostic tests performed, diagnosis, and treatment. RESULTS: Most horses were examined because of lameness (n = 72 [61%]) rather than poor performance (46 [39%]), but owner complaint was not significantly associated with age or body weight of the horse. The most common performance change was refusal or failure to turn properly around the first barrel (19/46 [41%]). The right forelimb (n = 57 [48%]) was most commonly affected, followed by the left forelimb (51 [43%]), the left hind limb (31 [26%]), and the right hind limb (25 [21%]). In 31 horses (26%), both forelimbs were affected, and in 6 (5%), both hind limbs were affected. The most common musculoskeletal problems were forelimb foot pain only (n = 39 [33%]), osteoarthritis of the distal tarsal joints (17 [14%]), suspensory ligament desmitis (15 [13%]), forelimb foot pain with distal tarsal joint osteoarthritis (11 [9%]), and bruised feet (10 [8.5%]). In 81 (69%) horses, the affected joint was treated with intra-articular medications. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that in horses used for barrel racing that are examined because of lameness or poor performance, the forelimbs are more likely to be affected than the hind limbs, with forelimb foot pain and osteoarthritis of the distal tarsal joints being the most common underlying abnormalities. PMID- 16313046 TI - [First trimester nuchal translucency thickness in normal fetuses in the Bulgarian population]. AB - Nuchal translucency (NT) measurement between 11-14 weeks of gestation is an effective method of ultrasound screening for chromosomal fetal anomalies. The aim of the present study is to construct normal reference ranges of NT thickness for the Bulgarian population. The choice of cut-off NT thickness above which an invasive procedure should be offered is discussed. PMID- 16313047 TI - [Evaluation of level of expression of p16 (INK4a) in endocervical lesions of uterine cervix]. AB - The incidence of malignant and premalignant endocervical glandular lesions is increasing. Several research groups have recently used the protein p16 (INK4a) as a possible diagnostic biomarker of cervical cancer. The objects of study were to evaluate whether the specificity of p16 (INK4a) expression in is sufficient for distinguishing normal, dysplastic and neoplastic endocervical epithelium. MATERIALS AND METHODS: Paraffin-embedded samples of diagnostic biopsies and surgical materials were used. Control group included biopsy samples from 10 patients with normal findings. We examined 20 samples with different pathologic endocervical lesions. Monoclonal antibody against p16 was used (DAKO). RESULTS: In control samples we did not find any p16 (INK4a)-positive cells. Overexpression of p16 (INK4a) was detected in samples of cervical dysplasia and invasive carcinomas. CONCLUSIONS: Overexpression of the protein p16 (INK4a) is typical for dysplastic and neoplastic epithelium of cervix uteri. p16 (INK4a) may be regarded as a supplementary test for routine diagnostics of cervical lesions. PMID- 16313048 TI - [Prognostic significance of Bishop's pelvic score for the continuity and the final outlet of induction of labor]. AB - Aim of the present mixed prospective and retrospective study is to determine the prognostic significance of the pelvic score of Bishop for the continuity and the final outlet of induction of labor and to determine the optimal score in the beginning of the induction related with a successful outlet. Based on the given results the authors conclude that the pelvic score of Bishop has a good prognostic value for the continuity and the final outlet of induction of labor with a various values of the separate parameters. Pelvic score > or = 5 is related with a shorter and more successful inductions of labor. PMID- 16313049 TI - [Impact of inherited thrombophilia on the development of some pregnancy complications]. AB - AIM: To assess the clinical significance of inherited thrombophilia [IT] for the development of some pregnancy complications. MATERIAL AND METHODS: The incidence of the following factors was studied in 97 pregnant women with pregnancy complications and in 103 controls: R506Q mutation encoding Factor V Leiden [FVL] synthesis, Prothrombin G20210A mutation, T677 methylenetetrahydropholate reductase mutation [MTHFR], 4G/4G polymorphism of the plasminogen activator inhibitor [PAI 4G/4G]. Among 97 patients in the group studied 39 had early onset severe preeclampsia [PE], 14--placental abruption [AP] without PE, 18- intrauterine growth restriction [IUGR] without PE, 12--stillbirth [SB] without PE, 14--habitual spontaneous abortions [HSA]. The control group included 103 clinically healthy pregnant women with at least one previous uneventful pregnancy, without history of thromboembolic disorders. In addition, patients with severe PE with and without IT were compared regarding g. a. and birthweight at delivery and intrauterine fetal loss rate. DNA analysis was performed according to internationally accepted standards. Pregnancy outcomes were ascertained from hospital records. Statistical significance (p < 0.05) was assessed by means of Student's t-test. RESULTS: FVL mutation was found in 23.7% (23/97) of the patients from the studied group and in 5.8% (6/103) of the controls. Prothrombin G20210A carriers were 11% (11/97) of the studied and 3.8% (4/103) of the controls, while with PAI 4G/4G polymorphism they were 30.9% (30/97) and 14.5% (15/103) respectively. MTHFR T677 was not more frequent in the studied group (8.2%) compared to the control one (29%). Eight of the patients (9.6%) were carriers of more than one mutation. In 22 cases with early onset severe PE and IT gestational age and birthweight at delivery were lower than in the cases with severe PE without IT while intrauterine fetal loss rate did not differ significantly between the two groups. CONCLUSIONS: Inherited thrombophilia is found more frequently in women with pregnancy complications like PE, IUGR, AP, SB, HSA. The incidence of homozygous MTHFR T677 is not higher in these cases. IT worsens the prognosis of severe PE. The diagnosis of IT is important since anithrombotic therapy has to be considered to protect the mother and the fetus. PMID- 16313050 TI - [Misoprostol and induction of labour]. AB - Misoprostol (Cytotec) is a synthetic analogue of PGE1, not yet registered for Obstetric practice. Vowing to its low cost and high efficacy there is marked interest towards this medicament. MATERIALS AND METHODS: 47 patients with induction of labour are distributed into two groups: Gr.A (n = 25), receiving Misoprostol and Gr.B (n = 22) with Oxytocin in perfusion. RESULTS: Protocol of treatment with PGE, includes 50 microg per os every 2 hrs. and in Gr. B standard solution of Oxytocin 5 IU. Before the beginning of induction Bishop Score and NST for 20 mins. is evaluated. The interval induction--delivery is much shorter in Gr.A, with a median of 479.00 +/- 105.44 mins., in comparison with 727.00 +/- 88.87 mins. in Gr.B. Uterine tachysystole is registered more frequently in Gr.A- 8%, with 4.54% in Gr.B. DISCUSSION: With receivers of Misoprostol, 76% deliver within 24 hrs. of induction. And there is no significant difference in the recorded monitor data for FDS and Apgar score of the neonate as well as the method of delivery. CONCLUSION: Oral Misoprostol is an effective method of induction of delivery with low Bishop Score. But there is a lack of sufficient data regarding the optimal regime and safety and with incorrect dosing may lead to unacceptable high incidence of complications. PMID- 16313051 TI - [Use of low molecular weight heparin (Clexane) together with selective COX-2 inhibitor (Dynastat--once or twice per day)]. AB - A follow-up of forty women was carried out. The patients having endured a gynecological operation (laparohysterectomy) were divided into two groups: group A--treated with Dynastat (40 mg/24h) and group B--treated with Dynastat (80mg/24h) which was combined in both groups with Clexane. The hematological and hemostatic parameters were evaluated during the early postoperative period--the first 24 hours. The women were followed until they were discharged from hospital. No significant changes of these laboratory parameters were noticed and the clinical stability of the patients in both groups was unaffected. The results showed that Dynastat/Clexane combination could be applied in the clinical practice as it proved great extend of safety for usage after gynecological operations. PMID- 16313052 TI - [Suppression of hyperprolactinemia in pituitary microadenoma with cabergoline (Dostinex)]. AB - First choice therapy of microprolactinoma is drug treatment with dopamine agonists. Cabergoline is widely accepted in clinical practice as a first line therapy for tumor-induced pituitary hyperprolactinemia. This study assessed serum prolactin levels, tumor size and adverse events in 22 women treated with cabergoline for one year (mean age 43.5 +/- 6.6 years). Serum prolactin levels changed from a baseline mean of 1417 +/- 347 UI/L to 489 +/- 102 UI/L at study end (in the normal range). Tumor size reduction to tumor disappearance was found in 18 women (from a mean of 7.2 mm to 5.5 mm), and was absent in 4 participants. Adverse events were reported in 2 women. In conclusion, the excellent therapeutic efficacy and the lack of adverse events with Cabergoline promotes its use as a first line therapy of hyperprolactinemia due to pituitary microadenoma. PMID- 16313053 TI - [Treatment of life-threatening bleeding in obstetrics and gynaecology --NovoSeven (recombinant factor VIIa)]. AB - The study offers the results from the application of recombinant factor VIIa (NovoSeven) in 19 patients with obstetric or gynaecological problems showing severe haemorrhage of non-surgical origin, which could not be controlled by standard therapy (surgery and resuscitation). Haematological and haemostaseological values as well as final outcome are being monitored. The role of the drug and its special indications in complex urgent treatment of life threatening bleeding are being discussed. Conclusions. The use of NovoSeven (recombinant factor Vlla) on time and when indicated guarantees rapid and effective haemostasis. Thus laparotomy or relaparotomy can be avoided. If no effect is observed after the application of NovoSeven indicates bleeding from a major blood vessel, demanding laparotomy and surgery. The use of the drug leads to reducing the incidence of blood derivatives transfusions and (being a recombinant product)--no risk of transmissive infections. PMID- 16313054 TI - [Ovarian and endometrial neoplasms and the use of hormone replacement therapy]. AB - Our research work summarize our and foreign Experience in the field of hormone replacement therapy and its use in endometrial and ovarian cancers. We look at the effects of the treatment on coronary heart diseases, venous thromboembolism, breast cancer and colorectal cancer. Results and examinations from other centers and clinics were included in our study treating the endometrial and ovarian cancers and the hormonal replacement therapy. Our aim was to work out guidelines and rules on prescribing hormone replacement therapy in clinical practice especially in oncogynecological malignancies. PMID- 16313055 TI - [Contraception in women with diabetes mellitus]. AB - Contraception in women suffering of diabetes mellitus is an important question mainly due to it has been proven that pregnancy outcome both for the fetus and for the mother depends on glycemic control before conception right to delivery. That's made planning of future pregnancy mandatory for patients with diabetes mellitus. Patients are advised for contraception until optimization of metabolic control has been achieved or until complete and contemporary treatment of the diabetes complications has been fulfilled. Incorrect contraceptive method could extend metabolic disorders and to reinforce vascular complications of the diabetes. Choice of contraception depends on the aim: short lasting contraception aiming on future pregnancy planning or long lasting contraception aiming on family planning. In women with diabetes mellitus it is extremely important to take into consideration such factors as type of the diabetes, its lasting, degree of metabolic compensation, presence of diabetic complications, body-mass index of the patient, presence of risk factors for cardiovascular diseases and future pregnancy planning. In cases when pregnancy is planning it has been preferred local contraception, such as condoms, diaphragms. In cases of nullipara or in women with plenty of partners, condoms are method of choice. In women who gave birth intrauterine device is such a method. In women with diabetes mellitus type I (insulin dependent diabetes mellitus) it is possible to use hormonal contraception depending on patient's request or on medical indications only in cases when diabetes has less than 15 years duration and microangiopathic complications and other vascular risk factors lack. Combined hormonal contraceptive preparations have to contain less than 30 microg ethinylestradiol and gestagen of "third" or "fourth" generation. This contraception has to be prescribed together with insulin dosage correction and demands on strict metabolic control of the diabetes and body weight. When the combined preparations are contraindicated it could be suggested progestagenic oral hormonal contraception if gynecologic contraindications lack and if this not leads to menstrual disturbances. In women with diabetes mellitus type II (non insulin dependent diabetes mellitus) combined hormonal contraceptives has not to be used, because they could provoke clinical manifestation and deteriorate progress of the diabetes mellitus. PMID- 16313056 TI - [Herpes simplex virus infection in the newborn]. AB - Herpes simplex virus (HSV) infection is a rare disease, but with serious sequelae in the newborn. Despite active treatment with high dose Acyclovir, prognosis on psychomotor development remains unfavorable. A short review of the recent literature about clinical forms, diagnosis, treatment, and disease prophylaxis is presented. PMID- 16313057 TI - [Cancer of the cervix uteri--exophytic form, prominating through the vaginal introitus of a 52-year old patient]. AB - The authors describe a rare case of carcinoma of the cervix uteri with exophytic accretion, prominating through the vaginal introitus of a 52-year old patient. They make the conclusion that accurate, precise and early diagnostics is necessary as well as an experienced operative team because during surgery there might be surprises, disturbing its normal course and outcome. PMID- 16313058 TI - [Magnetic resonance imaging for evaluation of neural chord defects of the fetus]. PMID- 16313059 TI - Technical report # 39: cold chain guidance for medicinal products: maintaining the quality of temperature-sensitive medicinal products through the transportation environment. Parenteral Drug Association. PMID- 16313060 TI - Leydig cells, thyroid hormones and steroidogenesis. AB - Leydig cells are the primary source of androgens in the mammalian testis. It is established that the luteinizing hormone (LH) produced by the anterior pituitary is required to maintain the structure and function of the Leydig cells in the postnatal testis. Until recent years, a role by the thyroid hormones on Leydig cells was not documented. It is evident now that thyroid hormones perform many functions in Leydig cells. For the process of postnatal Leydig cell differentiation, thyroid hormones are crucial. Thyroid hormones acutely stimulate Leydig cell steroidogenesis. Thyroid hormones cause proliferation of the cytoplasmic organelle peroxisome and stimulate the production of steroidogenic acute regulatory protein (StAR) and StAR mRNA in Leydig cells; both peroxisomes and StAR are linked with the transport of cholesterol, the obligatory intermediate in steroid hormone biosynthesis, into mitochondria. The presence of thyroid hormone receptors in Leydig cells and other cell types of the Leydig lineage is an issue that needs to be fully addressed in future studies. As thyroid hormones regulate many functions of Sertoli cells and the Sertoli cells regulate certain functions of Leydig cells, effects of thyroid hormones on Leydig cells mediated via the Sertoli cells are also reviewed in this paper. Additionally, out of all cell types in the testis, the thyrotropin releasing hormone (TRH), TRH mRNA and TRH receptor are present exclusively in Leydig cells. However, whether Leydig cells have a regulatory role on the hypothalamo-pituitary thyroid axis is currently unknown. PMID- 16313061 TI - A treatise on hazards of endocrine disruptors and tool to evaluate them. AB - Hormones mediate a major part of our essential physiological functions. Both endogenous and exogenous compounds and their metabolites are known to act through hormone receptors leading to regulation of endocrine function. The endogenous ligands that control reproductive functions are generally steroids such as 17beta estradiol, androgens, progesterone, pregnenolone and glucocorticoids. However, exogenous compounds that are structurally and functionally similar gain entry into animals including humans through the diet or by occupational exposures, causing endocrine disruption. In the recent decade, there is a lot of apprehension about the so-called "endocrine disruptors" which are wide spread in the environment, mainly due to unrestricted human activity. These compounds of anthropogenic or natural origin mimic the action of sex hormones and can interfere with the endocrine system. It has been hypothesized that environmental exposure to synthetic estrogenic chemicals and related endocrine active compounds may be responsible for malformations in the male reproductive tract, crytorchidism, hypospadias, decrease in sperm counts, decreased male reproductive capacity and even testicular cancers. The increasing concern in both public and scientific communities about these abnormalities have prompted the initiation of epidemiological studies to not only identify, but to also analyze the short and long term effects of endocrine disruptors. As a result, a number of assays have been developed and are undergoing validation aimed at high throughput screening of chemical agents that disrupt endocrine activity. This review consolidates the findings of epidemiological studies, particularly in relation to male reproductive disorders and brings to light the various types of in vitro and in vivo models that are available for tiered testing of suspected compounds. PMID- 16313062 TI - FSH, the neglected sibling: evidence for its role in regulation of spermatogenesis and Leydig cell function. AB - The role of follicle stimulating harmone(FSH) in male reproductive function remains a matter of debate although recent evidences strongly suggest a role despite the controversies that arose following the results obtained with FSH-beta null mice and observations from human FSH receptor mutations. This review summarizes the recent developments of our understanding on the role of FSH in male reproduction. Specifically the results obtained with FSH-beta and FORKO null mice are be discussed in light of our observations employing active and passive neutralization of endogenous FSH in rodents and primates along with other studies. On the basis of results obtained employing a variety of models it can be conclude unequivocally that FSH regulates Leydig cell function and is essential for normal quantitative spermatogenesis. PMID- 16313063 TI - Proteins implicated in sperm capacitation. AB - Spermatozoa after being deposited in the female reproductive tract spend a considerable time in this foreign environment prior to fertilization of the oocyte. Chang and Austin independently observed1,2 that this time spent by the spermatozoa in the female tract is not consequential but a necessary event in the life cycle of the male gamete, and Austin2 first called this maturation period of spermatozoa as 'Capacitation'. Ever since, attempts have been made to understand and unravel the molecular mechanism of capacitation. Based on the results obtained so far, it is clear that capacitation is guided by novel signal transduction pathways influencing varied aspects of spermatozoa. Capacitation could be, thus, defined as the cumulative molecular, cellular and physiological changes that occur in spermatozoa in the female reproductive tract to achieve the final competence to fertilize the oocyte. This review is structured so as to first understand the key features of capacitation and then to survey the players which bring about these changes during capacitation. PMID- 16313064 TI - Does human sperm nuclear DNA integrity affect embryo quality? AB - Abnormalities in the male genome are a clear potential reason for post fertilization failure. Male infertility may arise due to high levels of loosely packaged chromatin and damaged DNA. The achievement of a correct chromatin packaging level is essential for successful fertilization. The chromatin contained in the nuclei of mammalian spermatozoa is an extremely compact and stable structure. The reports on mammalian spermatozoa indicate that available volume is insufficient to contain sperm chromatin packed in nucleosome like structure and thus is organized in a special way. Different unique properties of sperm DNA like high degree of inertness and stability, absence of transcription, replacement of somatic histone by protamine etc have made the study of sperm chromatin more interesting. Increased levels of sperm nuclear DNA damage exist in infertile men with abnormal sperm parameters (i.e. concentration, motility and morphology), and various assay techniques have been developed to evaluate sperm chromatin maturity/DNA integrity. These assays are based on the facts that defects in chromatin structure have been shown to lead to increased DNA instability and sensitivity to denaturing stress. DNA integrity in the sperm is essential for the accurate and successful transmission of genetic information. Importance of sperm DNA has also become more obvious in the context of assisted reproductive techniques. While recent advances in assisted reproductive technologies have made possible and practical for many infertile men to become father, the risk of transmission of genetic mutation to the offspring, however, still remains. Further research is necessary to devise techniques for identification and selection of sperm with undamaged DNA for ICSI or to remove sperm with damaged DNA from the semen sample to improve the pregnancy outcome in ICSI. PMID- 16313065 TI - Current trends in evaluation of sperm function: in vitro selection and manipulation of male gametes for assisted conception. AB - With increasing medical utilization of assisted reproductive technology (ART), scientists and clinicians have been able to study extensively multiple cell functions operating synchronously and flawlessly during the events preceding, before and after fertilization. Critical evaluation of the functional status of spermatozoa for in vitro techniques such as sperm-mucus interaction, acrosome reaction status, sperm-zona pellucida binding and penetration tests, hyaluronic acid binding assay, and computer assisted semen analysis etc. can direct a male partner of an infertile couple to more aggressive forms of treatments. In vitro selection of functionally competent sperm cells is a pre-requisite for successful outcome in in vitro fertilization or in intracytoplasmic sperm injection (ICSI). Direct injections of acrosome-intact spermatozoa into oocyte during ICSI bypassing the normal events of sperm oocyte interaction and fusion events have raised concerns with regard to fertilization abnormalities and genetic issues. The present communication briefly reviews the sperm function tests with emphasis on its correlation with fertility outcome, and the currently employed sperm selection and manipulation procedures which may have implications in assisted conception programs. PMID- 16313066 TI - Male contraception: expanding reproductive choice. AB - The development of steroid-based oral contraceptives had revolutionized the availability of contraceptive choice for women. In order to expand the contraceptive options for couples by developing an acceptable, safe and effective male contraceptive, scientists have been experimenting with various steroidal/non steroidal regimens to suppress testicular sperm production. The non-availability of a long-acting androgen was a limiting factor in the development of a male contraceptive regimen since all currently tested anti-spermatogenic agents also concurrently decrease circulating testosterone levels. A combination regimen of long-acting progestogen and androgen would have advantage over an androgen-alone modality since the dose of androgen required would be much smaller in the combination regimen, thereby decreasing the adverse effects of high steroid load. The progestogen in the combination regimen would act as the primary anti spermatogenic agent. Currently, a number of combination regimens using progestogen or GnRH analogues combined with androgen are undergoing trials. The side effects of long-term use of androgens and progestogens have also undergone evaluation in primate models and the results of these studies need to be kept in view, while considering steroidal regimens for contraceptive use in men. Efforts are also being made to popularize non-scalpel vasectomy and to develop condoms of greater acceptability. The development of contraceptive vaccines for men, using sperm surface epitopes not expressed in female reproductive tract as source, still requires considerable research efforts. PMID- 16313067 TI - Perspectives of contraceptive choices for men. AB - Apart from condoms and vasectomy, which have several limitations of their own, no other methods of contraception are available to men. Various chemical, hormonal, vas based and herbal contraceptives have been examined and few of them have reached the stage of clinical testing. Promising leads have been obtained from testosterone buciclate/undecanoate, alone or in combination with levonorgestrel butanoate or cyproterone acetate, RISUG, an injectable intravasal contraceptive and a few herbal products, particularly the seed products of Carica papaya. It is feasible that an ideal male contraceptive, that meets out all the essential criteria will be made available to the community in the near future. PMID- 16313068 TI - Experimental hyperthyroidism-induced oxidative stress and impairment of antioxidant defence system in rat testis. AB - Short-term hyperthyroidism, induced by daily administration of T3 (20 microg/100g body weight) for one, three, and five days consecutively, modulates oxidative stress and antioxidant defence parameters in mitochondrial and postmitochondrial fractions of testis in adult rats. Alteration in antioxidant defences along with oxidative stress parameters in testis by thyroid hormone was found to be associated with a decline in the number of sperms and disturbances in histoarchitecture of seminiferous tubules in the testes; the results indicated that induced hyperthyroid state altered testicular physiology by influencing antioxidant defence system of testes. PMID- 16313069 TI - Effect of estradiol on expression of cytoskeletal proteins during spermatogenesis in testis of sexually mature rats. AB - Earlier studies had shown that long term treatment with estradiol arrests spermatogenesis in adult male rats, at a dose of 0.1 mg/kg/day. The present study was therefore undertaken to ascertain the causes underlying the reduction in sperm counts by administering estradiol for a short term, at the dose of 0.1 mg/kg/day. Estradiol valerate was injected at a dose of 0.1 mg/kg/day, for a period of 10 days to one group of adult male rats, which were administered saline for 12 days prior to estradiol injection, and sacrificed after 22 days. The control group was administered saline for 22 days. The sera were analyzed for testosterone and FSH levels. One testis of each male was immersion fixed for histology, and for immunohistochemistry of two testicular cytoskeletal proteins, vimentin and vinculin. The contralateral testes were used for analysis of vimentin and vinculin gene expression by reverse transcriptase polymerase chain reaction (RTPCR) and western blotting. Another group exposed to estradiol for 10 days was injected with bromodeoxyuridine (BrdU), at a dose of 100 mg/kg/day, to ascertain the effect on germ cell proliferation, and sacrificed 12 days later, while estradiol treatment was continued till sacrifice. BrdU, at a dose of 100 mg/kg/day was injected i.p. to a group of control rats treated with saline for 10 days, and sacrificed 12 days later. The testes from both groups were immersion fixed for immunohistochemical detection of BrdU. Histology of estradiol treated testis showed predominance of tubules with round spermatids with accumulation of lipid droplets in Sertoli cell cytoplasm and decreased cell height, whereas controls showed elongating spermatids. BrdU immunolocalization in the testis, irrespective of treatment, indicated its incorporation in deoxyribonucleic acid (DNA) suggesting that estradiol sustained germ cell proliferation. Both vimentin and vinculin could be immunolocalized to the testis. The testicular levels of vimentin and vinculin, quantified after western blotting, were unaffected. The testicular expression of vimentin and vinculin seen by RTPCR was also unaffected. The study suggested that estradiol induced reduction in sperm counts was not due to adverse effects on proliferation. The observed predominance of seminiferous tubules showing round spermatids, accumulation of lipid droplets as compared to controls suggested that reduction in elongated spermatids occurred through reduced spermiation and phagocytosis. The study also suggested that reduction in Sertoli cell height after short-term estradiol treatment was not due to reduced expression of vimentin and vinculin, which could be maintained by estradiol. However, reduction in Sertoli cell height could have been due to suppression of FSH and testosterone, implicated in the polymerization of vimentin and organization of vinculin, two cytoskeletal proteins involved in inter-Sertoli or Sertoli-germ cell junctions. The study suggested that disorganization of Sertoli cell cytoskeleton and reduction in the volume of Sertoli cells could be an important factor for reduced efficiency of spermatogenesis after exposure to estrogenic molecules. PMID- 16313070 TI - Germ cell death and their removal during initial stages of testicular ischemia and cryptorchidism: a comparative analysis. AB - Germ cell death and their removal from the seminiferous epithelium are common in the affected testis in conditions of unilateral ischemia or cryptorchidism; the similarities and differences, however, have not been studied between these two conditions. The present study was designed to examine the severity of the effect on testicular germ cells during the initial stages of both ischemia and cryptorchidism, which have significant implications on the restoration of fertility following surgical repair. Complete absence of spermatids was observed following 12 hr of ischemia as compared to 7 days of cryptorchidism. Germ cell removal in either case was in the direction of lumen to basement membrane leaving only a single layer of cells by 24 hr of unilateral ischemia as compared to 15 days of cryptorchidism. Levels of intratesticular testosterone was found lower in cryptorchidism (7 days) but not in ischemia till 24 hrs. Giant cells frequently observed in cryptorchid testis were absent in the ischemic seminiferous epithelium. There was a gradual increase in the number of apoptotic and non viable cells; the latter was more than 95% by 24 hr of ischemia. In contrast, approximately 85% testicular cells were nonviable till 15 days of cryptorchidism. The 1c peak representing the population of haploid cells was significantly reduced in cryptorchidism (7 days), while the peak was completely abolished by 24 hr of ischemia. Rise in the levels of oxidative stress in the affected testis was observed identically during the initial stages. These findings indicate that coupled with the rise in tissue oxidative stress, the number of apoptotic/nonviable germ cells was alarmingly high (> 80%) by 15 days of cryptochidism or 24 hr of ischemia. Restoration of complete spermatogenesis following surgical repair may not be possible in such cases because of these acute adverse effects. PMID- 16313071 TI - Male infertility: Y-chromosome deletion and testicular aetiology in cases of azoo /oligospermia. AB - The spermatogenesis locus azoospermia factor (AZF) in Yq11 has been delineated into three microdeletion intervals designated as AZFa, AZFb andAZFc. AZFc is the most frequently deleted region. We have studied 270 male infertile patients for various genetic disorders associated with infertile phenotype. In this study, we have presented results of our studies on Y-chromosome deletions, chromosomal abnormalities (Klinefelter syndrome) and histology of testis with the objective of seeing whether there were cases of gonosomic mosaicism and a causal correlation between the genetic disorder; and testicular aetiology could be drawn. In all the 13 cases of Y-chromosome microdeletion, AZFc region and DAZ gene were deleted, while no case of AZFa deletion was detected. This result was at variance with other reports from India, where a considerable fraction of cases showed deletion in AZFa region of the Y-chromosome. Both Y-deleted and non-Y deleted cases revealed heterogeneous testicular phenotype with comparable severity. This disparity among testicular phenotype in cases with known genetic aetiology and even in cases of unknown aetiology can be attributed to different genetic backgrounds and effect of modifiers. Since male infertility is a multifactorial disorder, the contributions of environmental and occupational insults may not be underestimated. PMID- 16313072 TI - Effect of aqueous leaf extract of Azadirachta indica on the reproductive organs in male mice. AB - Effect of oral administration (50, 100, and 200 mg/kg body weight/day, for 28 days) of aqucous leaf extract of neem (Azadirachta indica) on the male reproductive organs of the Parkes (P) strain mice was investigated. The treatment had no effect on body weight and the reproductive organs weight. In treated mice, testes showed both normal and affected seminiferous tubules in the same sections; the affected seminiferous tubules showed intraepithelial vacuolation, loosening of germinal epithelium, marginal condensation of chromatin in round spermatids, occurrence of giant cells, mixing of germ cell types in stages of spermatogenesis and degenerated appearance of germ cells. In severe cases, the tubules were lined with Sertoli cells only, Sertoli cells and rare germ cells, or with Sertoli cells and several germ cells but without cellular association patterns. Also, the frequency of affected seminiferous tubules in testes of the extract-treated mice was significantly higher than the controls, though this remained unaffected in mice treated at 50 mg/kg body weight of the extract. Doses at 50 or 100 mg/kg body weight of neem leaf extract did not cause appreciable alterations in histological appearance of the epididymis, while a dose of 200 mg/kg body weight caused marked alterations both in histological appearance and the level of sialic acid in the duct. The treatment also had adverse effects on motility, morphology, and number of spermatozoa in the cauda epididymidis, level of fructose in the seminal vesicle, and on litter size. After 42 days of withdrawal of the treatment, the alterations induced in the reproductive organs recovered to control levels. Our results suggested that treatment with neem leaf extract caused reversible alterations in the male reproductive organs of P mice. PMID- 16313073 TI - Sperm motility inhibiting activity of a phytosterol from Alstonia macrophylla Wall ex A. DC. leaf extract: a tribal medicine. AB - The role of methanolic extract and n-butanol fraction of A. macrophylla leaves was investigated on the forward motility of goat spermatozoa. The methanol extract (600 micro/g/ml) and one n-butanol fraction (Fraction A; 100 microg/ml) showed marked inhibition of sperm forward motility, tested by microscopic and spectrophotometric methods. Approximately, 50-60% of the spermatozoa lost their motility when treated with 600 microg/ml of methanol extract or 100 microg/ml of Fraction A. The Fraction A at 400 microg/ml concentration showed complete inhibition of sperm forward motility at 0 min. The inhibitory activity increased with the increasing concentrations of the fraction. The motility inhibitory activity of the Fraction A was stable to heat treatment at 100 degrees C for 2 min. The compound showed high inhibitory effect in the pH range 6.7-7.6. Fraction A also showed high efficacy for inhibiting human sperm motility, assessed by the microscopic method. The phytochemical analysis of methanolic extract of A. macrophylla leaves revealed the presence of sterols, triterpene, flavonoid, alkaloid, tannin and reducing sugar, while the Fraction A contains beta sitosterol, a common phytosterol. The results demonstrate that Fraction A (beta sitosterol) is a potent inhibitor of sperm motility and thus it has the potential to serve as a vaginal contraceptive. PMID- 16313074 TI - [XXIV National Obstetrics Conference and National Conference on Obstetrics, Kiten, 9-12, June, 2005. Part I]. PMID- 16313075 TI - [When is the time for a cesarean according to the desire of the pregnant woman?]. PMID- 16313079 TI - [Frequency and indicatrions for cesarean section in the Obstetrics Clinic in the University Hospital of Obstetrics and Gynecology "Maichin Dom" in Sofia]. PMID- 16313080 TI - [Incidence and indications for cesarean section in the Obstetrics Clinic in the University Hospital of Obstetrics and Gynecology "Maichin Dom" in Sofia]. AB - The incidence and the indications for Cesarean section in the Obstetric Clinic of the University Hospital of Obstetrics and Gynecology were analysed and compared to a study carried out 20 years ago In 2003 - 2004 there were 2700 pregnant women treated in the clinic 558 had Cesarean section (incidence 20.7%). For comparison the incidence of Cesarean section in 1985 was 6.67% (160 Cesarean sections for 2096 patients) and for 1986 the incidence was 7.63% (131 Cesarean Sections for 1865 patients. The leading indication for the both studies was "previous Cesarean section" the indication "contracted pelvis" was the second and infertility and breech presentation were on the third and forth place. There was no change in the indications for Cesqarean section in the present study and the study carried out 20 years ago. There was a relative increase of the indications 'contracted pelvis' and 'increased age in a nulliparous patiet' compared to the previous years. PMID- 16313081 TI - ["Pro" and "Con" cesarean section on demand]. PMID- 16313082 TI - [Cesarean section for fetal distress--is the decision interval for delivery significant?]. AB - The recommended Interval between the decision to perform emergency cesarean section and the delivery itself is 30 minutes. There is not enough clinical evidence in maintenance of this opinion. AIM: to study the interval from the decision of the cesarean section for fetal distress to delivery itself, as well as to analyze fetal and mother's outcome. METHODS: Prospective 13-month-study for the period 2003-2004 in Delivery room "Majchin Dom" included observation of 125 women. Cesarean section is classified in 3 Categories--1st--deemed highly hazardous for mother or/and fetus (crush); 2nd--not directly hazardous for mother or/and fetus (emergency); and 3dt--not direct mother or/and fetal distress, but the delivery is necessary in a short period of time (urgency). The Interval and the Categories are comparable with the presence or absence of fetal distress (Apgar < 7, pH < 7.20). RESULTS AND DISCUSSION: Positive results are found in the 1st category when the C-section is performed in 30 minutes. There are significant differences between the three Categories--the percentage of fetal distress is growing up comparable to prolongation of the time interval. Prolongation of the time interval for more than 60 minutes lead to poor fetal outcome. PMID- 16313083 TI - [Sectio caesarea in extremely low birth weight newborn babies--modern tendency or objective necessity]. AB - Obstetrician-gynecologists in a team with neonatologists contributed to saving of many children with extremely low birth weight /ELBW/, increasing many times SC frequency, considering it to be a sparing method for successful delivery, favorably influencing the outcome for the newborn. Many authors put emphasis on the better results however others point out reasons for a certain level of skepticism, assuming that survivability should not be the only criteria. Thus our objective was to find out the role of the abdominal delivery on the favorable outcome for ELBW newborn babies. MATERIAL AND METHODS: The study is retrospective, including 40 deliveries of children weighing from 500 to 999 g, at the Clinic in Obstetrics at Dr. Georgi Stranski University Hospital--Pleven, for a period of 5 years /2000-2004/. The newborn babies were divided into two groups depending on the method of delivery: I group--per vias naturales /PN/--31 cases- 77.5% II group--per SC--9 cases--22.5%. RESULTS AND DISCUSSION: Attention was paid in our study to the most frequently asked question about the survivability of the newborn babies until their discharge, neonatal diseases and mortality rate, respectively. Survivability rate in the first group was 32.2% compared to 66.6% for the newborn babies per SC. The main reasons for mortality in the newborn babies are massive brain hemorrhages--70% of the cases, respiratory insufficiency--25% and extreme immaturity--5%. CONCLUSIONS: 1. Operative delivery is an objective necessity as a method improving the final favorable outcome for ELBW newborn babies. 2. The intensive neonatal cares accompanied by precise application of surfactant sharply increase the survivability rate and reduce diseases and mortal rate of ELBW newborn babies. 3. The preventive cares in the maternity consultation rooms could become a landmark for reducing the number of pre-term deliveries. PMID- 16313084 TI - [CS by request--a reasonable alternative?]. AB - The doctor-patient relation has been changing as a whole together with the evolution of society. The right of choice should play a main role in female public health as it represents a fundamental social and constitutional law. The growing rate of caesarean sections in Stara Zagora, Bulgaria and in the world are analyzed and the organizational factors having an impact on it are shown. On-line polls among doctors and patients were used. A conclusion was drawn that it is better to have the indication "caesarean section by patient's request" rather than performing abdominal deliveries under fabricated indications. PMID- 16313085 TI - [Are we prepared for expectant mothers alone to choose the manner of delivery?]. PMID- 16313086 TI - [Application of hypnosis in obstetrics and gynecology]. PMID- 16313087 TI - [Nutrition of the newborn infant with very low birth weight with "non-natal" adapted milk and "nutrilon-premium" ]. PMID- 16313088 TI - Efficacy of ultrasound in removal of intraradicular posts using different techniques. AB - The efficacy of ultrasound in removing cast metal posts was evaluated in this in vitro study using one or two ultrasound units and ultrasonic vibration for various lengths of time. The crowns of 30 healthy maxillary canines were removed, the roots were embedded in acrylic resin blocks, and the root canals were treated endodontically. The canals were prepared and their impressions were taken with self-curing acrylic resin. After casting with copper-aluminum alloy, the posts were blasted with aluminum oxide and cemented with Panavia F resin cement. The specimens were divided into five groups. In groups I and II, only one ultrasound unit was used for 30 and 60 s, respectively; in groups III and IV, two ultrasound units were used simultaneously for 30 and 60 s, respectively; in group V, ultrasound was not used (control). Ultrasonic vibrations were applied with an Enac OE-5 ultrasound unit and an ST-09 tip. All samples were subjected to traction on an Instron machine (model 4444) at a cross-head speed of 1 mm/min. Analysis of the results revealed a statistically significant difference between the groups (ANOVA, P < 0.01); however the difference between groups II and IV was not statistically significant. The efficacy of ultrasound in removing intraradicular posts was confirmed, and the most effective technique was the use of two ultrasound units, independent of the length of time ultrasound was applied. PMID- 16313089 TI - A radiographic study of the relationship between technical quality of coronoradicular posts and periapical status in a Jordanian population. AB - A radiographic study was conducted to investigate the relationship between the technical quality of coronoradicular posts and periapical status. A total of 400 periapical radiographs, including 560 posts, of patients attending the Dental Department at Jordan University Hospital were scanned and studied. It was found that maxillary teeth were more frequently restored with posts (65.36%) than mandibular teeth (34.64%). Tapered posts accounted for 73.93% of the posts used. The ratio of the mean post length to crown length was 0.8, and that to root length was 0.45. The mean length of the remaining gutta percha apical to the end of the post was 6.22 mm. In addition, 25% of the posts deviated from the line of the root canal. Periapical radiolucency was evident in 53.93% of the assessed teeth. It is concluded that inadequate root canal treatment and coronoradicular posts are associated with increased prevalence of periapical radiolucency, and that general dental practitioners should be better trained in performing endodontic treatment and restoring endodontically treated teeth. PMID- 16313090 TI - Quantitative evaluation of axial wall taper in prepared artificial teeth. AB - The purpose of this study was to quantitatively evaluate the axial wall taper of prepared artificial teeth using a non-contact three-dimensional shape measuring system. A total of 54 artificial teeth prepared by pre-clinical dental students for complete cast restorations were evaluated. For quantitative analysis, five cross sections were computer-graphically placed perpendicularly to the z-axis. The surface coordinate values (x, y, z) of each cross section were converted into polar coordinate values (r, theta), which were then graphically rendered to a two dimensional plane. At four points, each 90 degrees from the distal center point of the cross section, the axial wall taper was quantitatively calculated using a formula based on the differences in radius between the highest and lowest positions of the cross sections of the prepared tooth. The average calculated taper was 5.8 degrees in the distal region, 21.7 degrees in the buccal region, 14.9 degrees in the mesial region and 12.5 degrees in the lingual region. These results suggest that the axial wall taper of prepared teeth can be quantitatively evaluated using this measuring system. PMID- 16313091 TI - Histological analysis of rat dental pulp tissue capped with propolis. AB - The aim of the present study was to assess the response of rat dental pulp to direct pulp capping with propolis. Flavonoid and non-flavonoid materials were purified from an ethanol extract of propolis obtained from South Sulawesi, Indonesia. A Class I cavity was prepared on the occlusal surface of the right maxillary first molar in Sprague Dawley rats. The dental pulp was exposed and then capped with a zinc oxide-based filler as a control (group I), or with propolis flavonoids (group II) or non-flavonoids (group III). The animals were sacrificed at week 1, 2 or 4, biopsy samples were obtained, and these were stained and viewed by light microscopy. The results showed that pulp inflammation occurred in groups I and III as early as week 1. No dentin bridge formation was seen in these groups. In contrast, there was no evident inflammatory response in group II at week 1. Mild and moderate pulp inflammation in this group occurred at 2 and 4 weeks after treatment, respectively. Partial dentinal bridge formation was seen in group II at week 4. Therefore, the present results suggest that direct pulp capping with propolis flavonoids in rats may delay dental pulp inflammation and stimulate reparative dentin. PMID- 16313092 TI - A single-retainer zirconium dioxide ceramic resin-bonded fixed partial denture for single tooth replacement: a clinical report. AB - This clinical report describes a treatment for the replacement of a missing mandibular anterior tooth using a cantilever single-retainer resin-bonded fixed partial denture (RBFPD), fabricated from zirconium dioxide (ZrO2) ceramic. No clinical complications were observed at the 2-year 6-month follow-up examination after placement of the ZrO2 ceramic RBFPD, and satisfactory functional and esthetic results were achieved. A treatment modality using a cantilever ZrO2 ceramic RBFPD is an alternative for single anterior tooth replacement. Further clinical studies are required to evaluate the long-term potential of cantilever single-retainer ZrO2 ceramic RBFPDs. PMID- 16313093 TI - Resolution of furcation bone loss after non-surgical root canal treatment: application of a peptidase-detection kit for treatment of type I endoperiodontal lesion. AB - Here, we report the management of a type I endoperiodontal lesion with furcation bone loss. A 59-year-old female attended our hospital with the chief complaint of mobility of tooth 46 and recurrent gingival swelling around the tooth. She previously received dental treatment from two dentists, but her condition did not improve. The tooth manifested the symptoms of typical periodontitis, such as gingival swelling, tooth mobility, pus discharge from the periodontal pocket and furcation bone loss. The tooth had no caries and the pulp reacted to an electric pulp test. Careful examination of the gingiva revealed traces of dental fistula. X-ray examination via a gutta percha inserted into the fistula revealed that furcation bone loss was associated with the periapical lesion. We diagnosed a type I endoperiodontal lesion, and applied Periocheck, a detection kit for peptidase-producing bacteria, to check for decreases in bacteria in the furcation and root canals. Soon after non-surgical root canal treatment, the condition of tooth 46 improved without periodontal treatment. After confirming a negative score with Periocheck, the root canal was filled. After 3 months, the furcation bone loss was on the way to recovery. These results indicate that proper diagnosis and confirmation of a decrease in root canal bacteria are important for treating endoperiodontal lesions. PMID- 16313094 TI - Case report of intra-osseous fibroma: a study on odontogenic and desmoplastic fibromas with a review of the literature. AB - Intra-osseous fibromas of the jaw are classified by origin. Intra-osseous odontogenic fibromas have odontogenic epithelia, while desmoplastic fibromas do not. However, it is often difficult to determine the odontogenic origin for central fibromas. Three subjects with a diagnosis of intra-osseous fibroma were examined. Case 1 was a 35-year-old man found to have a panoramic radiograph from the right premolar to the mandibular ramus in the mandible that exhibited multilocular radiolucency. Within the radiolucency, small-radioopaque bodies were observed. Case 2 was a 13-year-old female, in whom a panoramic radiograph from the left premolar to the molar in the mandible showed multilocular radiolucency. Case 3 was a 51-year-old female who exhibited a heart-shaped radiolucency in the panoramic radiograph of the left first molar area in the mandible. We also reviewed the literature for previously reported cases of intra-osseous odontogenic and desmoplastic fibroma. In 64 cases of intra-osseous odontogenic fibroma and 68 cases of desmoplastic fibroma we extracted data on age, sex, location, and radiographic findings. Based on the analysis of the reported literature cases, re-evaluation of the patients in our study revealed that case 1 could be classified as desmoplastic fibroma, while cases 2 and 3 were intra osseous odontogenic fibromas. PMID- 16313095 TI - Periodic exacerbation of gingival inflammation during the menstrual cycle. AB - Sex hormones are believed to be a risk factor for periodontitis because of their ability to proliferate specific periodontal microorganisms and affect host immunologic response. In this case report, gingival redness and swelling occurred during the menstrual cycle, although the patient maintained good oral hygiene during periodontal treatment. Medical history revealed that exacerbation of gingival inflammation corresponded to the menstrual cycle and occurred during the ovulation period, when estrogen levels are high. Mean bleeding index of the ovulation period (18.9%) showed higher levels than that during the menstrual phase (5.3%). This case indicates that frequent and effective maintenance should be provided while considering the influence of the menstrual cycle, as sex hormones may be involved in exacerbating gingival inflammation. PMID- 16313096 TI - [A role of nociceptin in the entrainment mechanism of the biological clock]. AB - The ORL1 receptor (nociceptin receptor), the fourth member of the opioid receptor family, is widely distributed throughout the central nervous system and mediates diverse physiological functions including pain, emotion, learning and memory. Nociceptin receptors are densely expressed in the suprachiasmatic nucleus, the principal pacemaker of circadian rhythms, and nociceptin inhibits light-induced phase advances (Allen et al, 1999); however, nociceptin's role in nonphotic entrainment has not been described. We used a small-molecule nociceptin receptor agonist capable of crossing the brain blood-brain barrier, and demonstrated that nociceptin acts as a nonphotic zeitgeiber. In this review, we describe a role for nociceptin in the biological clock and discuss the possibility of developing nociceptin receptor agonists for treatment of circadian rhythm disorders. PMID- 16313097 TI - [Clinical features of nicotine dependence]. AB - Nicotine dependence is characterized by weak dependence potential and less ability to produce psychotoxicity and social disturbance. A two-compartment model consisting of "dependence" and "dependence syndrome" was used to clarify clinical features of nicotine dependence. "Dependence" was defined by drug liking. "Dependence syndrome" was defined by a compulsion to take a drug, and drug induced pathological symptoms (withdrawal syndrome and acute disorders) and social disturbance. Nicotine produced a mild or the least degree of drug liking and withdrawal syndrome, without any significant social disturbance, or acute disorders. Thus, nicotine dependence differed from other forms of drug dependence in that nicotine was not associated with "dependence syndrome". This review also introduced other current topics of nicotine dependence. First, adolescence is regarded as a risk factor for the development of nicotine dependence, whereas the involvement of gender difference (female) in this respect is controversial. Secondly, many smokers feel difficulties in quitting smoking in spite of the weak dependence potential of nicotine, which is known as the "nicotine paradox". Several working hypotheses have been presented to explain this phenomenon. For example, nicotine has relatively strong conditioning effects and/or dependence liability compared with other drugs of abuse. However, further studies should be carried out to clarify clinical characteristics of the "nicotine paradox". PMID- 16313098 TI - Involvement of delta1-opioid receptors in the spatial learning impairment in streptozotocin-induced diabetic mice. AB - It is well accepted that diabetes leads to learning and memory impairment in humans and rodents. Because central delta-opioid receptors have important roles in learning processes, we investigated the involvement of delta-opioid receptors in the spatial learning impairment in streptozotocin (STZ)-induced diabetic mice by the Morris water maze test. The escape latencies to the platform were significantly increased in diabetic mice without changes in the ability to swim. The delta1/delta2-opioid receptor antagonist naltrindole (1 mg/kg/day, s.c.) slightly, but not significantly, reduced the escape latencies in diabetic mice. The selective delta1-opioid receptor antagonist 7-benzylidenenaltrexone (0.3 and 1 mg/kg/day, s.c.), but not the selective delta2-opioid receptor antagonist naltriben (0.3 and 1 mg/kg/day, s.c.), significantly reduced the escape latencies in diabetic mice. These antagonists had no effect on the escape latencies in non diabetic mice. The selective delta1-opioid receptor agonist [D-Pen2, D-Pen5] enkephalin (10 nmol/mouse/day, i.c.v.) significantly increased the escape latencies in both non-diabetic and diabetic mice. Based on these results, we suggest that the enhanced response to central delta1-opioid receptors in diabetic mice is involved, at least in part, in the spatial learning impairment in the Morris water maze test. PMID- 16313099 TI - [Control of craving for methamphetamine: development of scales for dependence and search for medicines for treatment]. AB - Methamphetamine dependence presents a serious problem not only for patients but also for society. Medical treatment has mainly targeted psychotic symptoms such as hallucination and delusion, and ignored the symptoms of craving, which are the major cause of dependence. Therefore, the risk of lapse into methamphetamine reuse remains very high. Although development of both medicines and programs for treatment of craving is needed, progress has been hampered by the lack of appropriate scales for assessing the severity of dependence and craving. On the other hand, recent breakthroughs in genomic sciences and molecular medicine have made it possible to investigate the molecular mechanisms underlying craving in animals. This paper reviews studies on the development of scales for assessing the severity of methamphetamine dependence and craving, together with recent data on candidate medicines for craving treatment in animals. The reliability and validity of the revised Addiction Severity Index -Japanese version (ASI-J) was confirmed after its administration to 100 drug abuse patients. The Craving Index was also newly developed, and its validity for prediction of relapse was confirmed. In animal experiments, fluoxetine, a selective serotonin reuptake inhibitor, was recognized as a candidate medicine for treatment of methamphetamine dependence. PMID- 16313100 TI - [Brain edema and aquaporin]. AB - Aquaporin is a membrane-bound water channel and highly related to the development and resolution of brain edema. We have detected mRNA and protein expression of aquaporin-4 in ischemic and traumatic brain injury. The aquaporin-4 expression is related to the resolution of brain edema rather than the evolution of the edema. We also detected the relation of aquaporin-1 expression and the evolution of hydrocephalus. Regulation of aquaporin function may be a new venue of treatment in brain edema. PMID- 16313101 TI - [Functions of microglia in immune mechanisms in the central nervous system]. AB - Microglia are only one type of professional immune cells in the central nervous system, functioning as scavenger cells that remove pathogens, and as antigen presenting cells that present brain-specific antigens to invading T cells. They also play a critical role in the pathophysiology of various CNS disorders as effector cells to destroy neural cells by secreting inflammatory cytokines, radicals, and glutamate. At the same time, they can protect neural functions by producing neurotrophic factors. Thus, to study the mechanisms for regulation of microglial functions may be useful for understanding the etiology and future therapeutic strategy for intractable neurological disorders. PMID- 16313102 TI - [Endogenous factors regulating neuronal apoptosis]. AB - Apoptosis and necrosis of neurons induced by glutamate and nitric oxide (NO) are associated with various disorders including hypoxic-ischemic brain injury, Alzheimer's disease and Parkinson's disease. In search of endogenous protective factors that inhibit NO-mediated glutamate neurotoxicity, we found that excitotoxicity is suppressed by certain neurotransmitters such as nicotinic acetylcholine and dopamine and growth factors such as NGF and BDNF. We recently purified and isolated a novel neuroprotective substance, which has been named 'serofendic acid', from a lipophilic fraction of fetal calf serum. Mass spectrometry and NMR spectroscopy revealed the chemical structure of serofendic acid (15-hydroxy-17-methylsulfinylatisan-19-oic acid) as a sulfur-containing atisane-type diterpenoid. Serofendic acid exhibited potent protective actions on cortical neurons against neurotoxicity of a NO donor as well as of glutamate, although it did not show appreciable influences on glutamate receptor-mediated responses in these neurons. Electron spin resonance analysis demonstrated that serofendic acid had no direct scavenging activity on NO radicals but was capable of inhibiting the generation of hydroxyl radicals. These findings suggest that serofendic acid is a low-molecular-weight bioactive factor that promotes survival of CNS neurons, probably through the attenuation of free radical-mediated insults. PMID- 16313103 TI - [Circadian rhythm study from anticipatory behavior to drug treatment]. AB - Precise, rhythmic, daily change of the internal milieu is a conspicuous feature of all living organisms. It affects temporal patterns of all kinds of behaviors during a day and deeply influences both the social structure and daily life of individual human beings. These daily variations arise from the internal circadian mechanisms. Three functions of the endogenous clock are discriminated as rhythm generation, entrainment to light-dark cycle and output from the clock. The endogenous clock is localized in the suprachiasmatic nucleus (SCN) in mammals. Recent papers demonstrated strong expression of clock genes such as Per1, Per2 and Per3 in the SCN. Circadian oscillation is basically regulated by the transcription/translation feedback system of the Per gene in mammals. As serotonin/antidepressant and GABA/benzodiazepine drugs affect the light and non light-induced entrainment, these drugs can regulate the circadian oscillation of clock genes and environmental stimuli-induced change of Per gene expression in the SCN. There are two main stimuli that entrain circadian rhythm, the light-dark cycle (LD) and restricted feeding. Light resets the circadian clock with induction of Per1 and Per2 gene in the SCN, the locus of a main oscillator. Mice were allowed access to food for 4 h during daytime (7 h in advance of feeding time) under LD or constant darkness. The peaks of mPer1 and mPer2 mRNA in the cerebral cortex and liver were advanced 6-12 h after 6 days of RF, whereas those in SCN were unaffected. The increase of mPer expression by RF treatment was observed in SCN-lesioned mice. The present results suggest that RF strongly entrained the expression of mPer and clock-controlled genes in the cerebral cortex and liver without affecting light-dependent SCN clock function. PMID- 16313104 TI - [Present status and research train of thoughts in integrative medicine on unstable plaque]. AB - The changing of atherosclerosis (AS) plaque from stable to unstable is closely related to the incidence of cardio-cerebrovascular events. So, to stabilize AS plaque become the hot point of the modern study on prevention and treatment of cardio-cerebrovascular diseases. The formation of AS plaque is a very long and complicated process with the participation of many factors. However, inflammation reaction plays an important role in the formation and rupture of unstable plaque. Traditional Chinese medicine (TCM) shows latent therapeutic advantages in stabilizing AS plaque with its characteristics of multi-way, multi-link, and multi-target point and all-sided treatment as well as its rather milder adverse reactions. The paper gives a comparatively systematic retrospection of study on unstable plaque treated by integrated Chinese and Western medicine conducted in recent years. Combined with the related topics conducted by the authors, they raise the theory of "unstable plaque is induced by toxinduced by toxin and stagnation", and introduce the study on the intervention of unstable plaque by removing toxic substances and activating blood flow. They believe that the explorative study can provide objective experimental and clinical basis for TCM therapeutic method in stabilizing AS plaque. PMID- 16313105 TI - [Effects of large dose of Astragalus membranaceus on the dendritic cell induction of peripheral mononuclear cell and antigen presenting ability of dendritic cells in children with acute leukemia]. AB - OBJECTIVE: To explore the effects of large dose of Astragalus membranaceus (Astragalus) on the dentritic cell (DC) induction in vitro and augumentation by peripheral mononuclear cell (MNC) and on antigen presenting ability of DC in children with acute leukemia. METHODS: Forty-four children with acute leukemia in complete remission stage were divided into two groups. Twenty patients in the Astragalus (90 g daily) group were treated with large dose of Astragalus (90 g daily) based on conventional chemotherapy for one month, while 24 patients in the control group received chemotherapy alone. MNC were extracted from peripheral blood by wall-sticking method and cultured with such cell factors as interleukin 4, gramulocyte macrophage colony stimulating factor, tumor necrosis factor-alpha for 7-8 days. Phenotype of DC was assayed by flow cytometry and antigen presenting ability of them was assayed by mixed lymphocyte reaction. RESULTS: There was no morphological difference in MNC induced DC between the two groups. The average number of DC in Astragalus group and control group was 4.4 x 10(6) / 2.5 x 10(6) MNC and 2.6 x 10(6) / 2.5 x 10(6) MNC, respectively, showing significant difference (P < 0.001). DC in Astragalus group could stimulate the proliferation of allogeneic lymphocytes strongly, showing significant difference when compared with that in the control group (P < 0.001). Conclusion Large dose of Astragalus could increase the DC induction of MNC and enhance the antigen presenting ability of DC in acute leukemia patients. PMID- 16313106 TI - [Clinical observation on jiangni hewei decoction in treatment of 45 patients with reflux esophagitis]. AB - OBJECTIVE: To observe the therapeutic efficacy of Jiangni Hewei Decoction (JHD) in treating reflux esophagitis (RE). METHODS: Seventy-five RE patients conformed with the inclusive criteria were randomly divided into the treated group (45 patients) and the control group (30 patients). Patients in the treated group received JHD orally, one dose per day, while those in the control group received Omeprazole, 20 mg once per day. The therapeutic course was 8 weeks. The efficacy on clinical symptoms, RE grades and scores determined by gastroscope were evaluated. RESULTS: Patients' symptoms were obviously improved after treatment of JHD and Omeprazole. The cured rate and the total effective rate in the treatedgroup were 37.8% (17 cases) and 84.5%, respectively, while in the control group were 36.7% (11 cases) and 83.3% respectively. The symptomatic score and the total score in both groups after treatment were obviously lowered (P < 0.01). Gastroscopic examination showed that the patients in the two groups with RE of 0 grade were significantly increased and no patients with RE of III grade. The healing rate and the total effective rate were 48.9% (22 cases) and 91.1% respectively in the treated group, and the corresponding numbers in the control group were 46.7% (14 cases) and 93.3% respectively. Significant difference was found in comparison of the gastroscopic figures in the same group before and after treatment (P < 0.01). The therapeutic effect of JHD on RE was similar to that of Omeprazole in aspects of improving clinical symptoms such as heartburn, acid regurgitation, chest pain, etc., and in curing oesophageal mucous inflammation (under gastroscope) and esophagitis. But the recurrence in the JHD group was obviously lower than that in the control group in 12 weeks after withdrawal of treatment (P < 0.05). Besides, no adverse reaction happened in the JHD group. CONCLUSION: JHD showed favorable therapeutic efficacy in treating RE. PMID- 16313107 TI - [Effects of acupuncture on the gastric motivity in patients with functional dyspepsia]. AB - OBJECTIVE: To observe the effects of the acupuncture on the gastric motivity in patients with functional dyspepsia (FD). METHODS: Ninety FD patients were randomly divided into 3 groups: the treated group, the control groups, 1 and 2, 30 patients in each group. Patients in the treated group received acupuncture treatment, those in the control group 1 and group 2 orally took Cisapride and Marzulene-s granule respectively. Changes in scores of symptoms, electrogastrographic figure, gastric emptying time estimated by B-ultrasonic examination and plasma level of motilin in the 3 groups before and after treatment were observed. RESULTS: Symptoms were obviously improved in the treated and control group 1 after treatment, which were better than those in the control group 2 (P < 0.01). Gastro-electric frequency and rhythm in the treated group and control group 1 were improved more significantly than those in before treatment (P < 0.01). Gastric emptying time and plasma motilin improved in the treated group and the control group 1 after treatment, when compared with before treatment, the difference was significant respectively (P < 0.05). CONCLUSION: Acupuncture could significantly improve the gastric motivity of FD patients. PMID- 16313108 TI - [Changes of Ag-NORs of T lymphocytes and hemorheological indexes in old patients with blood stasis syndrome]. AB - OBJECTIVE: To observe the changes of Ag-NORs of T lymphocytes and hemorheological indexes in old patients with blood stasis (BS) syndrome. METHODS: Ag-NORs of T lymphocytes and hemorheological indexes in 54 old patients with blood stasis syndrome in the BS group were determined and compared with those in the control group consisted of 36 healthy persons. RESULTS: Contents of Ag-NORs of T lymphocytes in the BS group and the control group were 5.857 +/- 0.396 and 6.111 +/- 0.541, respectively, showing significant difference (P < 0.05). Some abnormal hemorheological indexes were seen in the BS group, which were significant different to those in the control group (P < 0.05 ). Correlation analysis showed that RVP, ESR, K1 and K2 were negatively correlated with Ag-NORs of T lymphocytes (r = -0.273, -0.335, -0.301, -0.276 respectively; P < 0.01 or P < 0.05), while aggregation index was positively correlated with Ag-NORs of T lymphocytes (r = 0.389, P < 0.05). CONCLUSION: The lowered Ag-NORs of T lymphocytes and abnormal hemorrheological changes might be one of the important reasons for the lowered immunity of aged patients with BS syndrome. PMID- 16313109 TI - [Relationship between alcohol intake and TCM syndrome type in 1013 patients with cardio-cerebral diseases]. AB - OBJECTIVE: To explore the relationship between alcohol intake and traditional Chinese medicine Syndrome type in patients with cardio-cerebral diseases. METHODS: Study was conducted in 1013 selected inpatients with myocardial infarction (MI) or/and cerebral infarction (CI) by logistic multivariate regression analysis and comparison among groups. RESULTS: The scores of blood stasis syndrome showed a decreasing tendency along the increasing of alcohol intake, but it cannot prove that taking alcohol could prevent blood stasis. The scores of dampness-heat syndrome increased in patients who had chronically taken alcohol in large dose. CONCLUSION: Dampness-heat syndrome induced by alcohol is one of the basic mechanisms of cardio-cerebral diseases. Alcohol intake should be restricted. PMID- 16313110 TI - [Effect of extract of ginkgo bilboa leaf on early diabetic nephropathy]. AB - OBJECTIVE: To study the effect of extract of Ginkgo bilboa leaf (EGB) on renal lesions of early diabetic nephropathy (DN). METHODS: Sixty-eight patients with early DN were randomly divided into two groups, the control group (34 patients) and the treated group (34 patients). Patients in both groups received conventional therapy, while additional 9.6 mg EGB was orally taken by patients in the treated group, three times per day. The therapeutic course for both groups was three months. Indexes such as urinary microalbumin (mALB), alpha1 microglobulin (alpha1-MG), immunoglobulin (IgG), transferrin (TF), retinal binding protein (RBP) and N-acety-beta-D-glucosaminidase (NAG) before and after treatment between the two groups were compared respectively. RESULTS: Compared with before treatment, mALB, alpha1-MG, IgG, TF, RBP and NAG obviously decreased with significant difference in the treated group after treatment (P < 0.05). However, no significant decrease in the above-mentioned indexes in the control group (P > 0.05). CONCLUSION: EGB has the protective action on early DN. PMID- 16313111 TI - [Effect of arnebia root oil in promoting the tissue recovery of surface of wound and basic fibroblast growth factor (bFGF) mRNA expression]. AB - OBJECTIVE: To explore the molecular biological mechanism of Arnebia Root oil (AO) in promoting the recovery of surface of wound by observing basic fibroblast growth factor (bFGF) mRNA expression in the wound tissue and healing rate of the wound. METHODS: Patients in the observed group with their wound treated by AO and those in the control group treated by petrolatum gauze. The wound surface healing rate was estimated and bFGF mRNA expression was observed by RT-PCR. RESULTS: Endogenous bFGF mRNA expression existed in the wound surface of both groups, but its level in the observed group at any time point was obviously higher than that in the control group respectively, with significant difference in comparison of the gray density between the two groups (P < 0.05). The wound surface healing rate kept abreast with bFGF mRNA expression in wound tissues, so it was significantly higher in the observed group than that in the control group (P < 0.05). GAPDH gene, which was taken as a parameter for internal reference, expressed with a certain amount unchanged in different periods of healing (P > 0.05 ). CONCLUSION: AO shows obviously promotive action on bFGF, an important regulatory factor on wound healing, it might complete the recovery process by stimulating the increase of bFGF. PMID- 16313112 TI - [Clinical study of feiyanqing rectum condensed liquid in treating 36 cases of children syncytial viral pneumonia]. AB - OBJECTIVE: To evaluate the clinical efficacy and mechanism of Feiyangqin Rectum Condensed Liquid (FRCL) in treating children syncytial viral pneumonia. Methods Seventy-two patients were randomly divided into two groups, the 36 patients in the treated group were treated with FRCL, and the other 36 patients in the control group simply treated with Western medicine. Efficacy of treatment on clinical condition and some immune function (IgA, IgG, CD3, CD4) were observed. RESULTS: In the treated group, 28 patients were cured (77.8%), treatment was markedly effective in 4 patients (11.1%), effective in 2 (5.5%) and ineffective in 2 (5.6%), with the total effective rate of 94.4%. The corresponding number in the control group was 20 (55.6%), 7 (19.4%), 6 (16.7%), 3 (8.3%) and 91.7%, respectively. The cure rate in the treated group was obviously superior to that in the control group (P < 0.05). FRCL could improve serum IgA, IgG, CD3, CD4, and CD4/CD8, lower serum IgE, these indexes in the treated group were significantly different to those in the control group (P < 0.05). CONCLUSION: FRCL had the action in treating children syncytial viral pneumonia without any adverse reaction, one of its mechanisms might be related to its regulation on immune function. PMID- 16313113 TI - [Clinical study of SS syrup in treating xerostomia]. AB - OBJECTIVE: To compare the clinical efficacy of SS syrup, a Chinese medicine, and pilocarpine in treating patients with xerostomia. METHODS: Thirty-eight patients conformed to the inclusive criteria were randomly divided into two groups, they were treated by SS syrup (SS group) and pilocarpine (control group) respectively. Three indexes, i.e. questionnaire of dryness in mouth, total static salivary flow and dynamic salivary flow, before treatment, 1, 2 and 4 weeks after treatment were recorded and statistically analyzed. RESULTS: Significant difference was shown in the 3 indexes in the SS group between before treatment and 1, 2 and 4 weeks after treatment (P < 0.05), while in the control group significant difference was shown between before treatment and 1, 2, and 4 weeks after treatment except for total dynamic salivary flow after I weck treatment (P < 0.05). CONCLUSION: SS syrup, which has no adverse reaction, no contraindication, could be taken chronically, and shows good efficacy in improving the symptoms of xerostomia. PMID- 16313114 TI - [Effect of tongxinluo on endothelin-1 in the mini-swine model of acute myocardial infarction and reperfusion]. AB - OBJECTIVE: To evaluate the change of endothelin-1 (ET-1) in the mini-swine model of acute myocardial infarction (AMI) and reperfusion and the effect of Tongxinluo (TXL) on it, and to explore the possible mechanism of no-reflow. METHODS: Forty mini-swines were randomized into 5 groups: the model group, the small,middle and large dose of TXL groups and the sham-operated group, 8 in each group. The AMI reperfusion model was established by coronary ligation for 3 hrs followed with relaxation for 1 hr. Plasma ET-1 content before and after AMI, and after reperfusion was determined respectively by radioimmunoassay. The ET-1 mRNA expression in myocardial tissue of normal, ischemic and no-reflow area were respectively quantified by reverse transcription-polymerase chain reaction. RESULTS: (1) Compared with before AMI, levels of plasma ET-1 at the time points of 5 min and 3 hrs after AMI, 5 min and 1 hrs after reperfusion in the model group were significantly raised, showing an increasing tendency (all P < 0.01). But the increment in the middle and large dose of TXL groups were all lower than that in the model group (P < 0.05). (2) In the model and the TXL groups, levels of ET-1 in myocardial tissue of ischemic and no-reflow area were significantly higher than those in the normal area, and the increment in no-reflow area was higher than that in ischemic area (all P < 0.01). Compared with the model group, significant lowering of ET-1 in ischemic area was only shown in the middle and large dose of TXL groups (P < 0.01). (3) In the model and the TXL groups, ET-1 mRNA expression in ischemic area was significantly higher (all P < 0.01), while it in no-reflow area was significantly lower than that in the normal area respectively (all P < 0.01). The raised ET-1 mRNA expression in the middle and large dose TXL groups was significantly lowered when compared with that in the model group (P < 0.01). CONCLUSION: The endothelium injury might be one of the important mechanisms for no-reflow phenomenon. TXL might reduce the no-reflow by protecting endothelium cells. was significantly higher (all P < 0.01), while it in no-reflow area was significantly lower than that in the normal area respectively (all P < 0.01). The raised ET-1 mRNA expression in the middle and large dose TXL groups was significantly lowered when compared with that in the model group (P < 0.01). Conclusion The endothelium injury might be one of the important mechanisms for no-reflow phenomenon. TXL might reduce the no-reflow by protecting endothelium cells. PMID- 16313115 TI - [Study on the effect of combined therapy of Chinese and Western medicines and gene therapy (GT) on cartilage end-plate calcification in rabbits with cervical intervertebral disc regression]. AB - OBJECTIVE: To study the relationship between cartilage end-plate calcification (CEC) and cervical intervertebral discs regression (CIDR) in rabbits, and to study the inhibitory effect of combined therapy of Kanggu Zengsheng Capsule (KZC) ansforming growth factor-apsule (TGF-PLC) and igene therapy (GT) on CEC by measuring the thickness of CEC layer. METHODS: Thirty-five New Zeland rabbits of 4 months old were selected to establish cervical dynamic imbalance rabbit model for inducing CIDR (No disposal was given to rabbits in the normal control group). Seven months after operation, combined therapy of KZC and PLC were given, in doses calculated by body weight, to the modeled rabbits in the drug treated group with CEC of either superficial layer or full layer, twice a dantly by gavage for 30 successive days. While to those in the gene therapy group, the recombinant plasmid DNA with transforming growth factor-beta1 (TGF-beta1) was injected once their intervertebral discs (ID) of C(2-3) C(3-4) and C(4-5), 20 microl for each injection. One month later, all rabbits were sacrificed with periotic venous gas embolic method and their ID of C(4-5) (including partial body of the upper and lower vertebrae) was resected. The degree of CIDR was evaluated morphologically, and the thickness of CEC in rabbits was measured and compared between groups. RESULTS: Thickness of CEC in the model group, either of superficial layer or of full layer, was significantly more than that in the normal control group with significant difference. Both combined KZC and PLC therapy and gene therapy showed significant inhibitory effects on CEC in treating CIDR (P < 0.05). CONCLUSION: CEC is the initial factor of CIDR with highly positive correlation. Both combined therapy of KZC and PLC and gene therapy can significantly inhibit CEC. PMID- 16313116 TI - [Angiogenesis promoting effects of Chinese herbal medicine for activating blood circulation to remove stasis on chick embryo chorio-allantoic membrane]. AB - OBJECTIVE: To observe the angiogenesis promoting effects of clinical common used Chinese herbal medicines (CHM) for activating blood circulation to remove stasis on chick embryo chorio-allantoic membrane (CAM). METHODS: Chicken CAM model was established and mice blood serum containing different kinds of medicines, including Radix Peaoniae rubra, Radix Angelicae sinensis, Flos Carthami, Rhizoma Chuanxiong, Radix Salviae miltiorrhizae, Astragalus membranaceus, and their complex prescriptions, Danggui Buxue Decoction, Xuefu Zhuyu Decoction, Xiongshao Capsule, was applied on it respectively to observe the condition of angiogenesis 72 hrs after incubation. Besides, the normal saline group, blank serum group, blank group and basic fibroblast growth factor (bFGF) group were set up for control. RESULTS: All the CHM applied and bFGF had the CAM angiogenetic promoting effect, among them, Radix Salviae Miltiorrhizae and the three complex prescriptions showed better effects than the three negative control groups in capillary formation and count, with the efficacy similar to that of bFGF. The effect of complex prescriptions was superior to that of single herb except Radix Salviae miltiorrhizae. CONCLUSION: Radix Salviae miltiorrhizae, Danggui Buxue Decoction, Xuefu Zhuyu Decoction and Xiongshao Capsule have good angiogenesis promoting effect on CAM. This study elucidated, from a certain aspect, the mechanism of action of CHM on ischemic diseases, and unfolded the scientific evidence of applying complex prescription. PMID- 16313117 TI - [Study on effect of ginsenoside Rg1 in promoting myocardiac vascular endothelial cell regeneration through induction on bone marrow stem cell's migration and differentiation in rabbits of myocardial infarction]. AB - OBJECTIVE: To observe whether ginsenoside Rg1 could reduce the infarcted area and improve the heart function by path of promoting bone marrow stem cells differentiated to vascular endothelial cells (VECs). METHODS: Bone marrow was drawn from rabbit's ilium and labelled with red fluorochrome DiI, then it was transferred again into the rabbit's body. The rabbits was then made into myocardiac infarction model. The model rabbits were divided into the control group and the ginsenoside Rgl treated group (treated group). The infracted area at two weeks, and the left ventricular function at one and two weeks after infarction were determined respectively. The DiI positive cell rate of myelogenetic cells in ischmia area and CD31 positive cell rate of VECs were determined by confocal microscopy. Myocardial interstitial granulocyte colony stimulating factor(GCSF) levels during ischemia and reperfusion period were determined also. RESULTS: DiI positive rate of CD31 staining positive cells in the treated group was obviously increased, and the concentration of G-CSF in myocardium interstitial obviously increased, accompanied with obviously improving of heart function and obviously reducing of infarcted area. CONCLUSION: Ginsenoside Rgl could stimulate the G-CSF secretion in local myocardiac tissues, thus to induce bone marrow mononuclear cells migrate to myocadial tissue and further differentiate to VECs. The regeneration of endothelium cells show certain direct action in promoting capillary regeneration of infarcted myocardium tissue and maintaining the blood supply. PMID- 16313118 TI - [Protective effect of zuogui pill on ovarian autoimmune injury]. AB - OBJECTIVE: To probe the effect of Zuogui pill (ZGP), a Chinese compound recipe for tonifying Shen, on ovarian function in mice with premature ovarian failure (POF). METHODS: BALB/C female mice model of POF was established by multiple sites subcutaneous injection of ovarian antigen elicited with ovarian tissue of SD female rats, and treated with ZGP at different time points in the modeling, with prednisone as positive control. The levels of follicle-stimulating hormone (FSH) and estradiol (E2) in peripheral blood were measured with radioimmunoassay, and ovarian antibody (AoAb) was determined by enzyme linked immunosorbent assay. The mRNA expression of ovarian growth and differentiation factor-9 was detected with in situ hybridization. RESULTS: POF model mice manifested such abnormalities as increased FSH, decreased E2, and positive AoAb in peripheral blood, with lymphocytes infiltration in ovarian mesanchyma, reduction of GDF-9 mRNA positive oocytes, and decrease of growing and mature follicles. ZGP could reduce the increase of FSH, increase the level of E2, inhibit the production of AoAb, raise the GDF-9 mRNA positive cells of oocytes, increase the number of growing and mature follicles. The clinical efficacy was more significant in early stage than in advanced stage. CONCLUSION: ZGP can improve immune inflammatory injury of ovary, and shows therapeutic effect on POF. PMID- 16313119 TI - [Effect of radix bupleuri extract on cardiomyocyte apoptosis related genes expression in mouse infected with coxsackie virus B3m]. AB - OBJECTIVE: To explore the mechanism of Radix Bupleuri (RB) extract in regulating cardiomyocyte apoptosis. Methods The BALB/c mouse inoculated with the cardiomyocytophilic strain Coxsackie virus B3m (CVB3m) by peritoneal injection was adopted as viral myocarditis model. The experimental mice were divided into the normal control (A), the model (B) and the RB (C) groups. Mice in Group C were given 0.2 ml / 10 g of RB extract, 2 mg/ml in concentration by gastrogavage once daily, while equal volume of normal saline was given by gastrogavage to mice in Group A and B. All the mice were sacrificed on the 10th day after modeling. The cardiomyocyte apoptosis was detected by TDT/Dutp nick end labelling (TUNEL). The expression of Fas Bcl-2, and Caspase3 protein in myocardium was determined by immunohistochemistry and reverse-transcription polymerse chain reaction (RT-PCR). RESULTS: The apoptotic cell was not found in Group A, but Fas, Bcl-2 and Caspase3 were expressed in different degree. In Group B, a lot of apoptotic cells could be seen, with expression of Fas, Bcl-2 and Caspase3 stronger than those in Group A (P < 0.01). Compared with Group B, the apoptotic cardiomyocyte in Group C was significantly reduced (P < 0.01), with the expression of Fas and Caspase3 significantly lowered but expression of Bcl-2 increased slightly. CONCLUSION: RB extract could inhibit the cardiomyocyte apoptosis in animal with viral myocarditis, obviously down-regulate the expression of Fas and Caspase3, and slightly up-regulate Bcl-2 expression. PMID- 16313120 TI - [Therapeutic efficacy of lixue xiaoyin decoction in treating psoriasis and its effect on plasma edothelin]. AB - OBJECTIVE: To study the therapeutic efficacy of Lixue Xiaoyin Decoction (LXD) in treating psoriasis and its effect on plasma endothelin (ET). METHODS: Two hundred and twenty patients were divided into two groups. LXD was taken orally by the 118 patients in the treated group, while Compound Qingdai Capsule (CQC) taken orally by the 102 patients in the control group. The therapeutic efficacy was evaluated after 8 weeks of treatment. The ET content was determined by radioimmunoassay before and after treatment. RESULTS: The total effective rate was 73.72% in the treated group and 54.90% in the control group with significant difference (chi2 = 8.52, P < 0.01). The plasm ET level in patients was significantly higher than that in the healthy subjects. ET in both groups was all lowered after treatment, but the decrement was more obvious in the treated group, showing significant difference when compared with that in the control group (P < 0.01). CONCLUSION: LXD shows good therapeutic efficacy in the treatment of psoriasis. It can improve the microcirculation, inhibit the division of epithelial cells, promote the epidermic cell differentiation, and shows regulative effect on plasma ET. PMID- 16313121 TI - [Observation on short-term therapeutic effect of zhenggan recipe in treating liver cirrhosis with alpha-feto-protein]. AB - OBJECTIVE: To observe the therapeutic effect of Zhenggan Recipe (ZGR) in treating liver cirrhosis with abnormal Alpha-fetoprotein (AFP), and to explore its mechanism. METHODS: Sixty-four patients of liver cirrhosis with abnormal AFP were randomly divided into the treated group and the control group. All the patients received conventional combined Western medicinal treatment. Besides, additional ZGR was given to the patients in the treated group, one dose every day for orally taken in two times, 6 months as a therapeutic course. Changes in liver function (ALT, GGT, TBIL and ALB), AFP, T-lymphocyte subsets (CD4, CD4/CD8) and iconological examination were observed in the two groups before and after treatment to compared the efficacy. RESULTS: ZGR could effectively lower the AFP level (P < 0.01), and improve the levels of CD4 and CD4/CD8 (P < 0.01 or P < 0.05). CONCLUSION: ZGR can effectively lower the level of AFP, its mechanism might be related to the improving of cellular immune function of the patients. PMID- 16313122 TI - [Clinical observation of qingxue granules in treating post-nephrotransplantation erythrocytosis]. AB - OBJECTIVE: To observe the therapeutic efficacy of Qingxue Granules (QX) in treating postnephrotransplant erythrocytosis (PNTE). METHODS: Twenty patients were randomly divided into two groups according to the randomized table. QX was given to patients in the TCM treated group (QX group) and Enalapril given to patients in the Western medicine treated group (WM group), and the clinical efficacy in the two groups was observed. Results In the QX group, 3 patients got markedly effective, 2 effective, 2 improved, 1 ineffective, 1 dropped from the treatment, 1 absconded, with the total effective rate of 77.78%. The corresponding numbers in the WM group were 4, 2, 2, 1, 1 and 66.67%. There was no significant difference in comparison of the efficacies between the two groups (P > 0.05). There was no difference between the Intent-to-Treat population and Per protocol Pouplation after statistical management of lost cases. CONCLUSION: It has proved that QX has the same therapeutic effects as classic Western medical treatment in treating PNTE. The reliability and scientificity of QX was proved by Intent-to-Treat analysis. PMID- 16313123 TI - [Comments on Nigel Wiseman's "A Practical Sictionary of Chinese Medicine"--on Wiseman' s literal translation]. AB - Comments were made on the word-for-word literal translation method used by Mr. Nigel Wiseman in A Practical Dictionary of Chinese Medicine. He believes that only literal translation can reflect Chinese medical concepts accurately. The so called "word-for-word" translation is actually "English-word-for-Chinese character" translation. First, he made a list of Single Characters with English Equivalents, and then he replaced each character of Chinese medical terms with the assigned English equivalent. Many English terms thus produced are confusing. The defect of the word-for-word literal translation stems from the erroneous idea that the single character constitutes the basic element of meaning corresponding to the notion of "word" in English, and the meaning of a disyllabic or polysyllabic Chinese word is simply the addition of the meanings of the two or more characters. Another big mistake is the negligence of the polysemy of Chinese characters. One or two English equivalents can by no means cover all the various meanings of a polysemous character as a monosyllabic word. Various examples were cited from this dictionary to illustrate the mistakes. PMID- 16313124 TI - [Progress of treatment of mammary cancer]. PMID- 16313125 TI - [Progress of study on inhibitory effects of traditional Chinese herbs on growth factor induced proliferation of vascular smooth muscle cells]. AB - This paper sums up some studies in the last decade regarding the inhibitory effects of traditional Chinese herbs on growth factor induced proliferation of vascular smooth muscle cell (VSMC) via directly measuring the mRNA expression of its growth factors and the related receptors by electron microscope, immunohistochemistry, blot and hybridization in situ. PMID- 16313126 TI - [Progress of study on brain protective effect and mechanism of Polygonum multiflorum]. AB - This paper reviews the brain protective effect and mechanism of Polygonum multiflorum (PM), its extracts and active component, tetrahydroxystilbene glucoside (2,3,5,4'-tetrahydroxystilbene-2-O-beta-D-glucoside) published in recent decade. They have major effects as calcium channel antagonists, antioxidant, cholinomimetic drugs and cholinesterase inhibitors, as well as actions in regulating cell apoptosis and prolonging the ageing. The brain protective mechanism of PM is multi-target, multi-link and multi-way. Therefore, PM has great applicative value in prevention and treatment of senile neuropathies, such as Alzheimer's disease, Parkinson's disease and vascular dementia, etc. PMID- 16313127 TI - [Principle stages of scientific activity of the clinics for dietary nutrition affiliated to the Institute of Nutrition within 1930-2005 years]. AB - The article gives a brief account of activities of clinics for dietary nutrition affiliated to the Institute of Nutrition within 1930-2005 years. PMID- 16313128 TI - [Development of research in physiology and biochemistry of the digestion of the Institute of Nutrition of the Russian Academy of Medical Science for 1930-2005 years]. AB - This is a account of the main stages of formation and development of streamline of investigation of physiology and biochemistry of the digestion of the Institute of Nutrition the Russian Academy of Medical Science setting in 1930 years present time. PMID- 16313129 TI - [Principle stages of development and some scientific achievements of the laboratory for hygienic evaluation of food additives]. AB - In this paper is presented briefly the principle stages of researches, conducted in that laboratory during 1930 - 2005 years. PMID- 16313130 TI - [The study on consumption of calcium with food by the population of the Russian Federation]. AB - The analysis of actual nutrition of different groups of population of The Russian Federation, based on the data of The Russia Longitudinal Monitoring Survey 1994 2003, showed that more that 80% of the population consumes insufficient quantity of calcium. Taking into account low level of calcium consumption, bioavailability and its excretion, one can say, that calcium supply of the population of The Russian Federation is poor now. PMID- 16313131 TI - [Increase of nutritive value of poultry egg by of vitamin fortification of laid hens diets]. AB - Real content of vitamins A, E, B1 and B2 in hen's eggs from different poultry farms and personal farm varies in wide range and differs from the data of national food chemical tables. Elevated nutritive value eggs with maximal vitamin content may be obtained from hens fed diets with optimal vitamin level. Such way fortified one egg contributes 30% of recommended daily intake of vitamin E, about 10% of vitamins A and B2, 3% of vitamin B1. Intensification of egg yolk pigmentation by means of carotenoids additives to hens ration leads to their 10 fold increase comparing with the data of food chemical tables. Coloured egg provides for 5-10% of carotenoids adequate daily intake. PMID- 16313132 TI - [The influence of antioxidants on morphological structure of internal organs of rats in acute hypoxia]. AB - The results of pathomorphological research of internal organs of rats in hypoxical hypoxia and in the action of the antioxidant dihydroguersitine in acute hypoxia. Lt shows that DHQ causes favourable morphological changes in internal organs in acute hypoxia. PMID- 16313133 TI - [Effect of dietotherapy with food-stuffs for enteral nutrition on the dynamic of clinical and biochemic parameters in biliary tract diseases]. AB - It was investigated the influence of dietary therapy with enteral formula "Nutricomp AND Braum fiber" on clinical and biochemic parameters in biliary tract diseases. Also it was carried out the comparative estimation of the gallbladder motility in this contingent of patients in process of the standard food loading and the loading with the tested enteral formula (25 g carbohydrates). It was indicated that the including of enteral formula "Nutricomp AND Braum fiber" (209 g/day) in traditional hypocaloric diet promotes the decrease of cholesterol level and activity of alkaline phosphatase. The gallbladder motility was some smaller after the consuming of enteral formula "Nutricomp AND Braum Diabetes" compared with the standard food loading (25 g carbohydrates). PMID- 16313134 TI - [Influence of phospholipids on efficiency of dietary therapy and parameters of lipids metabolism in patients with hypertension]. AB - The estimation of efficiency complex dietary therapy with inclusion of phospholipids at patients with essential hypertension I and II stage. It was carried out(spent), that inclusion of 2 grams of lecithin in a diet for 4 weeks rendered positive influence on the clinical presentation of disease, condition antioxidant status in cellular membranes of erythrocytes and some biochemical indexes of lipid metabolism. PMID- 16313135 TI - [Theoretical and practical aspects of modern vitaminology]. AB - General subjects and achievement of modern vitaminology are discussed. The most impressive success in fundamental vitaminology during last three-four decades was achieved in following areas: elucidation of vitamins metabolic roles and molecular mechanisms in their action; development and improvement of high sensitive modern methods for analytical vitamins determination in biological objects and food; establishment of reliable criteria and methods for human vitamin status assessment; examination of physiological human vitamin requirements and establishment of Recommended Dietary Allowances (RDA) or Adequate Levels (AL) of their intake; elaboration of scientifical grounds for reasonable vitamins application in diseases prevention and treatment. In the area of applied vitaminology the most attention during last decades was paid to next subjects: regular broad vitamin status examination of representative groups of population; reasoning, development, realization and evaluation of broad scale measures for prevention of vitamin insufficiency and improvement of vitamin status of population; development and industrial production of vitamins, multivitamin-mineral preparations and various vitamin enriched food stuffs for the same purpose. Decrease of state financial support for fundamental studies and cessation of such a support for applied investigation make it necessary to organize nonstate commercial scientific-technologic center functioning on the requests of vitamin producers and consumers. PMID- 16313136 TI - Ground crew injuries and fatalities in U.S. commercial aviation, 1983-2004. AB - INTRODUCTION: Ground crew services are an essential part of airport operations. Injury hazards to ground crewmembers who are in close proximity to aircraft have not been well studied. The objective of this paper was to examine airport ground crew injuries and fatalities involving aircraft of commuter air carriers and major airlines. METHODS: Investigation reports for all ground crew injuries involving commuter and major airline aircraft that occurred at United States airports between 1983 and 2004 were obtained from the National Transportation Safety Board (NTSB) and analyzed to describe the immediate cause and pattern of injury occurrence. RESULTS: During the 22-yr study period, the NTSB recorded a total of 80 ground crew accidents involving landing, taxiing, or standing commercial airline aircraft, yielding an overall rate of 0.47 ground crew related accidents per 1 million aircraft departures. These accidents resulted in injuries to 98 ground crewmembers, including 21 fatalities. Two-thirds of the accidents took place as the aircraft was departing. Vehicular collisions with an aircraft made up 43% of accidents, 34% were caused by moving aircraft equipment such as propellers or nose gear, and 11% resulted from jet blasts or fires. CONCLUSIONS: intervention programs for airport ground personnel should emphasize the safe operation of the aircraft equipment and ground vehicles. Some of the injuries to ground crewmembers might be avoided through improved design of commonly used equipment. PMID- 16313137 TI - Metabolic responses to the seated calf press exercise performed against inertial resistance. AB - INTRODUCTION: Future in-flight strength training devices may use inertial resistance to abate mass and strength losses to muscle groups such as the triceps surae, which incurs pronounced deficits from space travel. Yet little data exist regarding physiological outcomes to triceps surae exercise performed against inertial resistance. Two sets of subjects were employed to note either blood lactate (La-) or net caloric cost responses to seated calf presses done on an inertial resistance ergometer. METHODS: Both sets of subjects performed 3 identical 3-set 10-repetition workouts. Blood La- measurements were made pre- and 5 min post-exercise. During workouts, breath-by-breath O2 uptake values were also recorded to help determine the net caloric cost of exercise. RESULTS: Compared to pre-exercise (mean +/- SEM) blood La- (2.01 +/- 0.08 mmol x L(-1)) values, post exercise (4.73 +/- 0.24 mmol x L(-1)) measurements showed a significant increase. Delta (post/pre differences) La- correlated significantly (r = 0.31-0.34) to several workout performance measures. Net caloric cost averaged 52.82 +/- 3.26 kcals for workouts; multivariate regression showed a subject's height, body mass, and body surface area described the variance associated with energy expenditure. CONCLUSIONS: Workouts evoked minimal energy expenditure, though anaerobic glycolysis likely played a major role in ATP resynthesis. Metabolic and exercise performance measures were likely influenced by series elastic element involvement of the triceps surae-Achilles tendon complex. Ergometer calf presses provided a high-intensity workout stimulus with a minimal metabolic cost. PMID- 16313138 TI - Muscle temperature and EMG amplitude and frequency during isometric exercise. AB - INTRODUCTION: While muscle temperature is known to vary with environmental temperature and the insulation provided by clothing, little has been published examining the interrelationships between the amplitude and frequency of the electromyogram (EMG), muscle tension, muscle fatigue, and muscle temperature. METHODS: Seven male subjects immersed their arms and legs in water at 24, 27, 34, and 37 degrees C for 20 min. Muscle temperature, strength (maximal voluntary contraction; MVC), endurance for a fatiguing contraction at 40% MVC, and EMG were assessed in the handgrip, biceps brachii, quadriceps, and gastrocnemius muscles. RESULTS: MVC was 44.8% lower for all muscles examined at the coldest muscle temperature. For all temperatures, the relationship between EMG amplitude and tension for brief isometric contractions was nearly linear; however, the increase in the amplitude of the EMG with muscle fatigue was reduced for the coldest muscle temperatures. The frequency components of the EMG and motor unit conduction velocity were largely unaffected by muscle tension but were inversely related to muscle temperature, with a 10 degrees C reduction in temperature resulting in a 32 Hz reduction in the center frequency. During fatiguing contractions at a tension of 40% MVC, the percent reduction in frequency was similar for all muscle temperatures, being reduced by about 20% from the beginning to the end of the contractions. DISCUSSION: EMG amplitude can be used to assess muscle use in most physiological conditions, but the frequency components of the EMG are so related to temperature as to make its use more restricted. PMID- 16313139 TI - EEG patterns associated with nitrogen narcosis (breathing air at 9 ATA). AB - INTRODUCTION: The narcotic effect of nitrogen impairs diver performance and limits dive profiles, especially for deep dives using compressed air. It would be helpful to establish measurable correlates of nitrogen narcosis. METHODS: The authors observed the electroencephalogram (EEG) of 10 subjects, ages 22-27 yr, who breathed air during a 3-min compression to a simulated depth of 80 msw (9 ATA). The EEG from a 19-electrode cap was recorded for 20 min while the subject reclined on a cot with eyes closed, first at 1 ATA before the dive and again at 9 ATA. Signals were analyzed using Fast Fourier Transform and brain mapping for frequency domains 0-4 Hz, 4-7 Hz, 7-12 Hz, and 12-15 Hz. Student's paired t-test and correlation tests were used to compare results for the two conditions. RESULTS: Two EEG patterns were observed. The first was an increase in delta and theta activity in all cortical regions that appeared in the first 2 min at depth and was related to exposure time. The second was an increase in delta and theta activity and shifting of alpha activity to the frontal regions at minute 6 of breathing air at 9 ATA and was related to the narcotic effects of nitrogen. DISCUSSION: If confirmed by studies with larger case series, this EEG pattern could be used to identify nitrogen narcosis for various gas mixtures and prevent the dangerous impact of nitrogen on diver performance. PMID- 16313140 TI - Multiple caffeine doses maintain vigilance during early morning operations. AB - INTRODUCTION: When used to counteract the effects of sleep deprivation, multiple doses of caffeine are typically ingested across an extended period of time. The goal of this study was to determine the optimal dose of caffeine for sustaining performance during sleep loss with administration of multiple doses. METHODS: There were 48 subjects (28 men, 20 women) who were randomly assigned to 1 of 4 groups (placebo, 50, 100, or 200 mg caffeine). After an overnight 8-h sleep period, subjects were required to remain awake for the ensuing 29 h. Control data were collected until 03:00 (Day 3), followed by three 2-h test blocks. At 03:00, 05:00, and 07:00 subjects chewed two sticks of gum (Stay Alerts chewing gum) containing caffeine or placebo. Six 10-min sessions on a version of the Psychomotor Vigilance Test (PVT) were completed during each 2-h test block. The Stanford Sleepiness Scale (SSS) was administered after each PVT. RESULTS: Lapses on the PVT were categorized as response times greater than 1, 3, or 5 s. Lapses in all categories significantly increased in the placebo group. Caffeine significantly reduced the number of lapses in a dose-related manner; and performance was maintained at baseline levels for the entire sleep loss period with multiple doses of 200 mg caffeine. There was a significant main effect for session on the SSS, the score increasing over time, but no significant differences between groups. DISCUSSION: These results indicate that a bi-hourly administration of 200 mg of caffeine maintains vigilance performance across a single night without sleep. PMID- 16313141 TI - Effects of ethnicity and gender on motion sickness susceptibility. AB - INTRODUCTION: Susceptibility to motion sickness (MS) is known to be affected by gender and ethnic origin, but whether gender and ethnicity are interacting is unknown. METHODS: We investigated MS development in healthy Caucasian subjects (n = 227), and in subjects of Chinese origin (n = 82). All subjects were exposed to nausea-inducing body rotations in a rotation chair, and rotated around the yaw axis for 5 x 1 min, while they were instructed to move their heads. Prior to rotation, subjects had to fill out a motion-sickness susceptibility questionnaire (MSSQ). Total rotation tolerance time (RT) was noted. Symptom ratings (SR) were performed at the beginning, and immediately after the end of each rotation, and 15 and 30 min later. RESULTS: The average RT was significantly higher in Caucasian (163 +/- 6 s) than in Chinese subjects (111 +/- 7 s) (F = 24.84, p < 0.0001). The adult MSSQ score was significantly lower in Caucasians (17.8 +/- 1.1) than in Chinese volunteers (24.2 +/- 2.1) (F = 6.05, p = 0.014). Maximal SR post rotation was similar in Chinese and Caucasian subjects. RT was highly predictable from the MSSQ scores, but separate for both genders. CONCLUSION: Susceptibility to MS is affected by both ethnic origin and by gender in a rather complex fashion. The most reliable prediction of RT can be based on the individual's history as assessed by the MSSQ. PMID- 16313142 TI - Sleep measurement in flight crew: comparing actigraphic and subjective estimates to polysomnography. AB - INTRODUCTION: Flight crew sleep is recognized as critical to alertness and safety, but few studies have compared the reliability of the different types of sleep monitoring that are commonly used. The present study compared actigraphic and subjective estimates of sleep to the gold standard, polysomnography (PSG). METHODS: In-flight (25 episodes) and layover sleep (21 episodes) of 21 flight crew were measured with PSG and actigraphy. Subjective reports were made 30 min after in-flight sleep episodes. Actigraphy data were analyzed at low, medium, and high activity thresholds. RESULTS: Actigraphic and subjective estimates of sleep duration correlated highly with PSG (range 0.84-0.95), regardless of sleep location or activity threshold. Mean differences were relatively small (-36-20 min), but the 95% confidence intervals of the differences were wide (+/- 71- +/- 103 min), particularly for subjective estimates (+/- 112 min). Actigraphic estimates of sleep efficiency and latency showed moderate to poor correlation with PSG values. Epoch-by-epoch comparisons showed the actigraph was sensitive to sleep (83-95%), but not very specific (34-62%). Kappa values indicated only weak agreement (< 0.4). DISCUSSION: For estimating mean sleep duration, both actigraphic and subjective estimates are sufficiently close to PSG values, but the amount of random error must be considered. Any single estimate may vary by more than 1 h from the mean difference. Neither actigraphy nor subjective estimates are suitable for estimating sleep efficiency and latency. Findings indicate that the performance of the actigraph is not altered in flight, other than the predictable effects associated with shorter, more disturbed sleep. PMID- 16313143 TI - Smoking cessation therapy and the return of aviators to flying duty. AB - Smoking cessation is an important part of every primary care physician's work. The importance of smoking cessation in the reduction of cardiovascular morbidity and mortality and the reduction of cancer incidence cannot be overstated. Various treatments have been established to encourage smoking cessation; these include group and individual psychological therapy, nicotine replacement in various forms, and drug therapy. The best-known drug used for smoking cessation is bupropion SR (Zyban). Smoking in aviators is not different than in the general population in terms of prevalence. Thus it is important for flight surgeons worldwide to be familiar with the magnitude of the problem and the available treatment options. Yet, it is also important for this community to become familiar with the relevance of this treatment to aviation and to recognize the limitations pertinent to flying personnel who are attempting to quit smoking. We present treatment options for smoking cessations and their limitations on flying personnel. PMID- 16313144 TI - Neutrophil degranulation response to 2 hours of exercise in a 30 degrees C environment. AB - INTRODUCTION: Evidence supports an interaction between neuro-endocrine responses to exercise and immune responses to exercise. We hypothesized that prolonged exercise in the heat would evoke a greater stress hormone response and a greater decrease in neutrophil degranulation [lipopolysaccharide (LPS)-stimulated elastase release] than when the same exercise was performed in thermoneutral conditions. METHODS: In counterbalanced order and separated by 7 d, 13 male cyclists cycled for 2 h at 62 +/- 3% VO2max (mean +/- SEM), with ad libitum water intake, on one occasion with heat (HOT: 30.3 degrees C, 76% RH) and on another occasion without ( CONTROL: 20.4 degrees C, 60% RH). Venous blood samples were collected at pre-, post-, and 2 h post-exercise. RESULTS: Exercising HR, rating of perceived exertion, rectal temperature, corrected body mass loss, and plasma cortisol at post- and 2 h post-exercise were greater during HOT. A marked neutrophilia was evident at post- and 2 h post-exercise with no difference between trials. LPS-stimulated elastase release per neutrophil decreased post exercise with no difference between trials (pre-exercise: HOT 189 +/- 20 and CONTROL 210 +/- 32; post-exercise: HOT 127 +/- 18 and CONTROL 136 +/- 29 fg x cell(-1)). There was no effect of exercise or trial on neutrophil CD11b expression (neutrophil activation index) or band cell percentage (neutrophil maturity index). CONCLUSIONS: Prolonged exercise results in a decrease in neutrophil degranulation that is unaffected by performing the exercise in hot conditions despite the increase in physiological stress. Additionally, these data suggest that the decrease in neutrophil degranulation after prolonged exercise is not associated with a change in neutrophil activation or maturity as previously suggested. PMID- 16313145 TI - Headache and altitude decompression sickness: joint pain or neurological pain? AB - INTRODUCTION: Exposure to reduced ambient pressure may result in decompression sickness (DCS). Headache is among the DCS symptoms encountered and is usually regarded as neurological DCS, which is traditionally classified as serious DCS. Since cranial sutures may be considered joints, it is possible that some headaches are actually joint pain and when associated with decompression sickness need not be neurological DCS. METHODS: Records were individually recovered from the Davis Hyperbaric Laboratory at Brooks City-Base, TX. Information was extracted using a detailed survey instrument. Possible joint pain headache cases were identified using three criteria: headache localized at a suture, normal neurologic exam, and resolution within 30 min of hyperbaric oxygen treatment. RESULTS: A total of 729 records documenting treatment for DCS were scrutinized. Of these, 70 cases of altitude DCS with headache were examined. Analysis, using the three criteria, showed 23% (16 cases) of altitude headache DCS symptoms could potentially be re-classified as joint pain. CONCLUSION: Generally, headache DCS is considered neurological DCS. However, since cranial sutures are joints, both histologically and functionally, and since DCS most commonly affects joints, headache DCS may, at times, be joint pain DCS. Indeed, retrospective data analysis suggests that this possibility exists. Such a reclassification from neurological to joint pain DCS would lessen the aeromedical impact of a DCS headache. PMID- 16313146 TI - Pressure-equalizing earplugs do not prevent barotrauma on descent from 8000 ft cabin altitude. AB - INTRODUCTION: The aim of this study was to evaluate the effect of pressure equalizing earplugs available in major airports and drugstores. No previous study has focused on preventing barotrauma using these earplugs. METHODS: Blinded and double-blinded, one type of pressure-equalizing earplugs (JetEars) was studied in 27 volunteers disposed to ear barotrauma. They acted as their own controls with an active earplug in one ear and a placebo earplug in the other ear at random. All were exposed to the same well-defined pressure profile for 1 h at 8000 ft, comparable to the environment in civil commercial air travel in a pressurized cabin. Satisfaction was assessed by questionnaire and objective results were evaluated prior to and after the pressure exposure by tympanometry and otoscopy using the Teed classification. RESULTS: The majority of the volunteers (78%) reported a pleasant noise-reducing feeling using the earplugs. However, 75% also experienced ear pain during descent. In comparing the middle ear pressure before and after pressurization, a decrease was found in ears with both active earplugs and placebo earplugs. No difference between the active and the placebo earplugs were found. Furthermore, after evaluation of the two groups of ears using otoscopy, no prevention of barotrauma was found. In fact, the ears using an active pressure-equalizing earplug scored significantly worse (p = 0.033). CONCLUSIONS: Feelings of noise reduction were reported, but no prevention of barotrauma could be demonstrated with the use of pressure-equalizing earplugs. Pressure-equalizing earplugs cannot be recommended in air travel for preventing ear barotrauma. PMID- 16313147 TI - Ethological indicators of isolated and confined teams in the perspective of missions to Mars. AB - INTRODUCTION: This study proposes an ethological overview of the social behavior of confined and isolated teams within three experimental chambers and at a polar base during long-duration campaigns. These were undertaken as analogous conditions of the mobility restraint and social constraint to which Mars teams would have to adapt. METHOD: The ethological method consisted of weekly observations and descriptions of the subjects' motor actions and spatial positions at dinnertime, then evaluating their levels of frequency in terms of distances, orientations, and dispersions, as well as facial expressions and body movements. RESULTS: Changes in social behavior indicate different adaptive strategies over time and according to the situation. In large or open areas, interindividual distances are constant. In reduced habitats, the frequency of personal distances decreases and the frequency of public distances increases with high levels of social distance and body mobility from the initial period to the final period. Increasing spatial dispersion and decreasing social orientations indicate low levels of sociality within the teams at the mid-period. Frequent collateral activities during long-duration confinement campaigns indicate high levels of stress in the team members. DISCUSSION: Such ethological indicators could be used in future interplanetary mission scenarios. PMID- 16313148 TI - You're a flight surgeon: aeromedical decisions in a case of optic disc drusen. PMID- 16313149 TI - You're the flight surgeon: mesangiocapillary glomerulonephritis (MPGN) with secondary polycythemia. PMID- 16313151 TI - Hand in glove? PMID- 16313152 TI - Here we make friends. PMID- 16313153 TI - On side. PMID- 16313154 TI - Travelling hopefully. PMID- 16313155 TI - The bigger picture. How do we contemplate tragedy and suffering? Our secular culture provides no answers. PMID- 16313156 TI - Community treatment. PMID- 16313157 TI - Lost in the system. PMID- 16313158 TI - Inclusive and empowering. PMID- 16313159 TI - Award caps Bastille's career. PMID- 16313160 TI - Ariel: articles from the ether (if you take it out of the box!). PMID- 16313161 TI - Document delivery made easy. PMID- 16313162 TI - Beware of roadblocks on the way to Ariel. PMID- 16313163 TI - Ariel--not just for academic libraries. PMID- 16313164 TI - Ariel cometh and Ariel goeth. PMID- 16313165 TI - DOCLINE update. PMID- 16313166 TI - Enzyme function of the globin dehaloperoxidase from Amphitrite ornata is activated by substrate binding. AB - Amphitrite ornata dehaloperoxidase (DHP) is a heme enzyme with a globin structure, which is capable of oxidizing para-halogenated phenols to the corresponding quinones. Cloning, high-level expression, and purification of recombinant DHP are described. Recombinant DHP was assayed by stopped-flow experiments for its ability to oxidatively debrominate 2,4,6-tribromophenol (TBP). The enzymatic activity of the ferric form of recombinant DHP is intermediate between that of a typical peroxidase (horseradish peroxidase) and a typical globin (horse heart myoglobin). The present study shows that, unlike other known peroxidases, DHP activity requires the addition of substrate, TBP, prior to the cosubstrate, peroxide. The presence of a substrate-binding site in DHP is consistent with a two-electron oxidation mechanism and an obligatory order for activation of the enzyme by addition of the substrate prior to the cosubstrate. PMID- 16313167 TI - Roles of dimerization domain residues in binding and catalysis by aminoacylase-1. AB - The aminoacylase-1/metallopeptidase 20 (Acy1/M20) family is the largest metallopeptidase family. Several crystal structures feature a metal-binding and a dimerization-mediating domain, both arranged in an extended open conformation. We have recently shown [Lindner et al. (2003) J. Biol. Chem. 278, 44496-44504] that in human Acy1 the invariant residues Glu147 and His206 from the metal-binding and the dimerization domain, respectively, are recruited to the active site from opposite dimer subunits. We hypothesized that, to facilitate this, formation of the binary complex is associated with domain closure, which would also position additional residues in the functional active site of Acy1. These would include two partially conserved dimerization domain residues: an asparagine (Asn263) and an arginine (Arg276) from the same subunit as His206 and Glu147, respectively. In this paper, we investigate the significance of the three dimerization domain residues of human Acy1 His206, Asn263, and Arg276 and, additionally, the nearby Asp274 for catalysis using site-directed mutagenesis. Enzyme complementation assays confirm the putative subunit allocations of these residues, and steady state kinetics support roles for all of them in catalysis but only involve the Arg276 in substrate-binding. The results are consistent with a model of the closed conformation for the structure of the related enzyme carboxypeptidase G2. This study demonstrates experimentally for the first time for a member of the Acy1/M20 family that several residues outside of the metal-binding domain are involved in binding and catalysis. PMID- 16313168 TI - Enzymatic generation of the chromopyrrolic acid scaffold of rebeccamycin by the tandem action of RebO and RebD. AB - During the biosynthesis of the fused six-ring indolocarbazole scaffolds of rebeccamycin and staurosporine, two molecules of L-tryptophan are processed to a pyrrole-containing five-ring intermediate known as chromopyrrolic acid. We report here the heterologous expression of RebO and RebD from the rebeccamycin biosynthetic pathway in Escherichia coli, and tandem action of these two enzymes to construct the dicarboxypyrrole ring of chromopyrrolic acid. Chromopyrrolic acid is oxidized by six electrons compared to the starting pair of L-tryptophan molecules. RebO is an L-tryptophan oxidase flavoprotein and RebD a heme protein dimer with both catalase and chromopyrrolic acid synthase activity. Both enzymes require dioxygen as a cosubstrate. RebD on its own is incompetent with L tryptophan but will convert the imine of indole-3-pyruvate to chromopyrrolic acid. It displays a substrate preference for two molecules of indole-3-pyruvic acid imine, necessitating a net two-electron oxidation to give chromopyrrolic acid. PMID- 16313169 TI - Prostate-cancer-associated I260M variant of DNA polymerase beta is a sequence specific mutator. AB - Studies show that 30% of 189 tumors sequenced to date express variants of the polymerase beta (pol beta) protein that are not present in normal tissue. This raises the possibility that variants of pol beta might be linked to the etiology of cancer. Here, we characterize the I260M prostate-cancer-associated variant of pol beta. Ile260 is a key residue of the hydrophobic hinge that is important for the closing of the polymerase. In this study, we demonstrate that the I260M variant is a sequence context-dependent mutator polymerase. Specifically, I260M is a mutator for misalignment-mediated errors in dipyrimidine sequences. I260M is also a low-fidelity polymerase with regard to the induction of transversions within specific sequence contexts. Our results suggest that the hinge influences the geometry of the DNA within the polymerase active site that is important for accurate DNA synthesis. Importantly, characterization of the I260M variant shows that it has a functional phenotype that could be linked to the etiology or malignant progression of human cancer. PMID- 16313170 TI - Using 2-aminopurine fluorescence to detect bacteriophage T4 DNA polymerase-DNA complexes that are important for primer extension and proofreading reactions. AB - The fluorescence of the base analogue 2-aminopurine (2AP) was used to probe bacteriophage T4 DNA polymerase-induced conformational changes in the template strand produced during the nucleotide incorporation and proofreading reactions. 2AP fluorescence in DNA is quenched by 2AP interactions with neighboring bases, but T4 DNA polymerase binding to DNA substrates labeled with 2AP in the templating position produces large increases in fluorescence intensity. Fluorescence lifetime studies were performed to characterize the fluorescent complexes. Three fluorescence lifetime components were observed for unbound DNA substrates as reported previously, but T4 DNA polymerase binding modulated the amplitudes of these components and created a new, highly fluorescent 10.5 ns component. Experimental evidence for correlation of fluorescence lifetimes with functionally distinct complexes was obtained by forming complexes under different reaction conditions. T4 DNA polymerase complexes were formed with DNA substrates with matched and mismatched primer ends and with A+T- or G+C-rich primer-terminal regions. dTTP was added to binary complexes to form ternary DNA polymerase-DNA nucleotide complexes. The effect of temperature on complex formation was studied, and complexes were formed with proofreading-defective T4 DNA polymerases. Complexes characterized by the 10.5 ns lifetime were demonstrated to be formed at the crossroads of the primer-extension and proofreading pathways. PMID- 16313171 TI - A synthetic peptide modeled on PDNF, Chagas' disease parasite neurotrophic factor, promotes survival and differentiation of neuronal cells through TrkA receptor. AB - The human parasite Trypanosoma cruzi, the agent of Chagas' disease, expresses a membrane-bound neuraminidase/trans-sialidase, also known as parasite-derived neurotrophic factor, PDNF, because it binds and activates nerve growth factor (NGF) receptor TrkA in neuronal cells. Here, we identify a 21 amino acid region (425GNASQNVWEDAYRCVNASATAN445) of PDNF that reproduces its neurotrophic activities. Synthetic peptide Y21, modeled on this sequence, induces survival and neurite outgrowth in primary dorsal root ganglion neurons. Y21 but not other PDNF based peptides promotes survival and neurite extension in TrkA-expressing but not in TrkA-deficient PC12 cells. Y21 also enhances phosphorylation of TrkA in PC12 cells and activation of Erk1/2 and Akt kinases with kinetics distinct from that of PDNF. In addition, Y21 stimulates phosphorylation of cAMP response element binding protein, CREB. Peptide Y21, therefore, reproduces several TrkA-dependent activities of PDNF and NGF. However, Y21 inhibits the binding of PDNF but not NGF to TrkA. Similarly, Y21 blocks PDNF- but not NGF-dependent phosphorylation of Erk1/2. These findings raise the possibility that Y21 reacts with a TrkA site required for the binding of PDNF but not NGF. The functioning of Y21 as TrkA agonist reproducing TrkA-dependent biological activities of PDNF should help elucidate the mechanism of PDNF activation of TrkA-expressing cells and the design of small drugs for the treatment of Chagas' and other neurodegenerative diseases. PMID- 16313172 TI - The Rv0805 gene from Mycobacterium tuberculosis encodes a 3',5'-cyclic nucleotide phosphodiesterase: biochemical and mutational analysis. AB - Mycobacterium tuberculosis is an important human pathogen and has developed sophisticated mechanisms to evade the host immune system. These could involve the use of cyclic nucleotide-dependent signaling systems, since the M. tuberculosis genome encodes a large number of functional adenylyl cyclases. Using bioinformatic approaches, we identify, clone, and biochemically characterize the Rv0805 gene product, the first cyclic nucleotide phosphodiesterase identified in M. tuberculosis and a homologue of the cAMP phosphodiesterase present in Escherichia coli (cpdA). The Rv0805 gene product, a class III phosphodiesterase, is a member of the metallophosphoesterase family, and computational modeling and mutational analyses indicate that the protein possesses interesting properties not reported earlier in this class of enzymes. Mutational analysis of critical histidine and aspartate residues predicted to be essential for metal coordination reduced catalytic activity by 90-50%, and several mutant proteins showed sigmoidal kinetics with respect to Mn in contrast to the wild-type enzyme. Mutation of an asparagine residue in the GNHD motif that is conserved throughout the metallophosphoesterase enzymes almost completely abolished catalytic activity, and these studies therefore represent the first mutational analysis of this class of phosphodiesterases. The Rv0805 protein hydrolyzes cAMP and cGMP in vitro, and overexpression in Mycobacterium smegmatis and E. coli reduces intracellular cAMP levels. The presence of an orthologue of Rv0805 in Mycobacterium leprae suggests that the Rv0805 protein could have an important role to play in regulating cAMP levels in these bacteria and adds an additional level of complexity to cyclic nucleotide signaling in this organism. PMID- 16313173 TI - Purification and characterization of a recombinant G-protein-coupled receptor, Saccharomyces cerevisiae Ste2p, transiently expressed in HEK293 EBNA1 cells. AB - The production of milligram quantities of purified, active, folded membrane protein from heterologous expression systems remains a general challenge due to intrinsically low expression levels, misfolding, and instability. Here we report the overexpression and purification of milligram quantities of functional Saccharomyces cerevisiae G-protein-coupled receptor, Ste2p, from transiently transfected human embryonic kidney 293 EBNA1 cells. Fluorescent microscopy indicates localization of Ste2p-GFP and Fc-Ste2p-GFP fusion receptors to the cell membrane. Up to 2 mg (approximately 10 pmol/million cells) of the Fc-Ste2p-GFP fusion and 1 mg of a Ste2p-Strep-TagII/(His)8-tagged version were purified per liter of culture following protein A-Sepharose and Talon metal affinity chromatography, respectively. Two distinct fluorescent labels, the hydrophobic 7 (diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM) and the more hydrophilic fluorescein-5-maleimide (FM), were individually attached to the C terminus of the alpha-mating factor ligand by addition of a reactive cysteine residue to produce active fluorescent pheromones. In vitro fluorescent ligand binding assays demonstrated that a high percentage of the recombinant purified receptor is correctly folded and able to bind ligand. KD values of 34 +/- 3 and 300 +/- 20 nM were observed respectively for the CPM- and FM-labeled ligands. These results combined with blue-shifted emission peaks and loss of fluorescent quenching observed for both fluorescent-labeled Cys alpha-factors when bound to receptor support a model in which the C-terminus of the ligand is packed in a hydrophobic pocket at the interface between the transmembrane and extracellular loop domains. Overall, we present an efficient system for recombinant production of milligram quantities of purified Ste2p in a biologically active form with applications to future structure and functional studies. PMID- 16313174 TI - Liprin phosphorylation regulates binding to LAR: evidence for liprin autophosphorylation. AB - The LAR transmembrane tyrosine phosphatase associates with liprin-alpha proteins and colocalizes with liprin-alpha1 at focal adhesions. LAR has been implicated in axon guidance, and liprins are involved in synapse formation and synapse protein trafficking. Several liprin mutants have weaker binding to LAR as assessed by yeast interaction trap assays, and the extents of in vitro and in vivo phosphorylation of these mutants were reduced relative to that of wild-type liprin-alpha1. Treatment of liprin-alpha1 with calf intestinal phosphatase weakened its interaction with the recombinant GST-LAR protein. A liprin LH region mutant that inhibited liprin phosphorylation did not bind to LAR as assessed by coprecipitation studies. Endogenous LAR was shown to bind phosphorylated liprin alpha1 from MDA-486 cells labeled in vivo with [32P]orthophosphate. In further characterizing the phosphorylation of liprin, we found immunoprecipitates of liprin-alpha1 expressed in COS-7 cells to incorporate phosphate after washes of up to 4 M NaCl. Additionally, purified liprin-alpha1 derived from Sf-9 insect cells retained the ability to incorporate phosphate in in vitro phosphorylation assays, and a liprin-alpha1 truncation mutant incorporated phosphate after denaturation and/or renaturation in SDS gels. Finally, binding assays showed that liprin binds to ATP-agarose and that the interaction is challenged by free ATP, but not by free GTP. Moreover, liprin LH region mutations that inhibit liprin phosphorylation stabilized the association of liprin with ATP-agarose. Taken together, our results suggest that liprin autophosphorylation regulates its association with LAR. PMID- 16313175 TI - Electrostatic switches that mediate the pH-dependent conformational change of "short" recombinant human pseudocathepsin D. AB - Human cathepsin D (hCatD) is an aspartic peptidase with a low pH optimum. X-ray crystal structures have been solved for an active, low pH (pH 5.1) form (CatD(lo)) [Baldwin, E. T., Bhat, T. N., Gulnik, S., Hosur, M. V., Sowder, R. C., Cachau, R. E., Collins, J., Silva, A. M., and Erickson, J. W. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 6796-6800] and an inactive, high pH (pH 7.5) form (CatD(hi)) [Lee, A. Y., Gulnik, S. V., and Erickson, J. W. (1998) Nat. Struct. Biol. 5, 866-871]. It has been suggested that ionizable switches involving the carboxylate side chains of E5, E180, and D187 may mediate the reversible interconversion between CatD(hi) and CatD(lo) and that Y10 stabilizes CatD(hi) [Lee, A. Y., Gulnik, S. V., and Erickson, J. W. (1998) Nat. Struct. Biol. 5, 866 871]. To test these hypotheses, we generated single point mutants in "short" recombinant human pseudocathepsin D (srCatD), a model kinetically similar to hCatD [Beyer, B. M., and Dunn, B. M. (1996) J. Biol. Chem. 271, 15590-15596]. E180Q, Y10F, and D187N exhibit significantly higher kcat/Km values (2-, 3-, and 6 fold, respectively) at pH 3.7 and 4.75 compared to srCatD, indicating that these residues are important in stabilizing the CatD(hi). E5Q exhibits a 2-fold lower kcat/Km compared to srCatD at both pH values, indicating the importance of E5 in stabilizing the CatD(lo). Accordingly, full time-course "pH-jump" (pH 5.5-4.75) studies of substrate hydrolysis indicate that E180Q, D187N, and Y10F have shorter kinetic lag phases that represent the change from CatD(hi) to CatD(lo) compared to srCatD and E5Q. Intrinsic tryptophan fluorescence reveals that the variants have a native-like structure over the pH range of our assays. The results indicate that E180 and D187 participate as an electrostatic switch that initiates the conformational change of CatD(lo) to CatD(hi) and Y10 stabilizes CatD(hi) by hydrogen bonding to the catalytic Asp 33. E5 appears to play a less significant role as an ionic switch that stabilizes CatD(lo). PMID- 16313176 TI - Early events in apomyoglobin unfolding probed by laser T-jump/UV resonance Raman spectroscopy. AB - Early events in the unfolding of apomyoglobin are studied with time-resolved ultraviolet resonance Raman (UVRR) spectroscopy coupled to a laser-induced temperature jump (T-jump). The UVRR spectra provide simultaneous probes of the aromatic side-chain environment and the amide backbone conformation. The amide bands reveal helix melting, with relaxation times of 70 and 16 micros at pH 5.5 and 4, respectively, in reasonable agreement with previously reported amide I' FTIR/T-jump relaxations (132 and 14 micros at pD 5.5 and 3). The acceleration at pH 4 is consistent with destabilization of the hydrophobic AGH core of the protein via protonation of a pair of buried histidines. The same relaxation times are found for intensity loss by the phenylalanine F12 band, signaling solvent exposure of the phenyl rings. There are seven Phe residues, distributed throughout the protein; they produce a global response, parallel to helix melting. Relaxation of the tryptophan W16 intensity also parallels helix melting at pH 5.5 but is twice as fast, 7 micros, at pH 4. The pH 5.5 signal arises from Trp 7, which is partially solvent-exposed, while the pH 4 signal arises from the buried Trp 14. Thus, Trp 14 is exposed to the solvent prior to helix melting of the AGH core, suggesting initial displacement of the A helix, upon which Trp 14 resides. All of the UVRR signals show a prompt response, within the instrument resolution (approximately 60 ns), which accounts for half of the total relaxation amplitude. This response is attributed to solvent penetration into the protein, possibly convoluted with melting of hydrated helix segments. PMID- 16313177 TI - A chemical strategy to manipulate the intracellular localization of drugs in resistant cancer cells. AB - A number of multidrug-resistant (MDR) cancer cells have been shown to have acquired an increased capacity to sequester weakly basic anticancer drugs in their lysosomes relative to drug-sensitive counterparts. In this report we have comparatively evaluated the concentrations of the anticancer agent daunorubicin (DNR) in intracellular compartments of drug-sensitive and MDR HL-60 cell lines, both of which do not express common efflux transporters such as P-glycoprotein at the plasma membrane. Our results suggest that lysosomal sequestration plays a significant role in the emergence of MDR since it effectively limits the drug's ability to interact with target molecules located in the nucleus. Using a series of weakly basic structural isomers with variable basicity, we illustrate that the magnitude of the pKa value correlates with the degree of lysosomal sequestration. Accordingly, a series of structurally modified forms of DNR with reduced basicity were synthesized, and their intracellular distribution was evaluated. Consistent with model compounds, derivatives of DNR with lowered pKa values showed visibly reduced lysosomal sequestration in two separate MDR cell lines. Collectively, this work highlights the importance of understanding the intracellular localization of drugs and proposes a rational strategy to manipulate it. PMID- 16313178 TI - The binding of W7, an inhibitor of striated muscle contraction, to cardiac troponin C. AB - W7 is a well-characterized calmodulin antagonist. It decreases the maximal tension and rate of ATP hydrolysis in cardiac muscle fibers. Cardiac troponin C (cTnC) has been previously implicated as the mechanistically significant target for W7 in the myofilament. Two-dimensional NMR spectra ({1H,15N}- and {1H,13C} HSQCs) were used to monitor the Ca2+-dependent binding of W7 to cTnC. Titration of cTnC x 3Ca2+ with W7 indicated binding to both domains of the protein. We examined the binding of W7 to the separated domains of cTnC to simplify the spectral analysis. In the titration of the C-terminal domain (cCTnC x 2Ca2+), the spectral peaks originating from a subset of residues changed nonuniformly, and could not be well-described as single-site binding. A global fit of the cCTnC x 2Ca2+ titration data to a two-site, sequential binding model (47 residues simultaneously fit) yielded a dissociation constant (Kd1) of 0.85-0.91 mM for the singly bound state, with the second dissociation constant fit to 3.40-3.65 mM (> or = 4 x Kd1). The titration data for the N-terminal domain (cNTnC x Ca2+) was globally fit to single-site binding model with a Kd of 0.15-0.30 mM (41 residues fit). The data are consistent with W7 binding to each domain's major hydrophobic pocket, coordinating side chains responsible for liganding cTnI. When in muscle fibers, W7 may compete with cTnI for target sites on cTnC. PMID- 16313179 TI - Cytotoxicity of bovine seminal ribonuclease: monomer versus dimer. AB - Bovine seminal ribonuclease (BS-RNase) is a homologue of bovine pancreatic ribonuclease (RNase A). Unlike RNase A, BS-RNase has notable toxicity for human tumor cells. Wild-type BS-RNase is a homodimer linked by two intermolecular disulfide bonds. This quaternary structure endows BS-RNase with resistance to inhibition by the cytosolic ribonuclease inhibitor protein (RI), which binds tightly to RNase A and monomeric BS-RNase. Here, we report on the creation and analysis of monomeric variants of BS-RNase that evade RI but retain full enzymatic activity. The cytotoxic activity of these monomeric variants exceeds that of the wild-type dimer by up to 30-fold, indicating that the dimeric structure of BS-RNase is not required for cytotoxicity. Dimers of these monomeric variants are more cytotoxic than wild-type BS-RNase, suggesting that the cytotoxicity of the wild-type enzyme is limited by RI inhibition following dissociation of the dimer in the reducing environment of the cytosol. Finally, the cytotoxic activity of these dimers is less than that of the constituent monomers, indicating that their quaternary structure is a liability. These data provide new insight into structure-function relationships of BS-RNase. Moreover, BS-RNase monomers described herein are more toxic to human tumor cells than is any known variant or homologue of RNase A including Onconase, an amphibian homologue in phase III clinical trials for the treatment of unresectable malignant mesothelioma. PMID- 16313180 TI - Crystal structure of homoserine transacetylase from Haemophilus influenzae reveals a new family of alpha/beta-hydrolases. AB - Homoserine transacetylase catalyzes one of the required steps in the biosynthesis of methionine in fungi and several bacteria. We have determined the crystal structure of homoserine transacetylase from Haemophilus influenzae to a resolution of 1.65 A. The structure identifies this enzyme to be a member of the alpha/beta-hydrolase structural superfamily. The active site of the enzyme is located near the end of a deep tunnel formed by the juxtaposition of two domains and incorporates a catalytic triad involving Ser143, His337, and Asp304. A structural basis is given for the observed double displacement kinetic mechanism of homoserine transacetylase. Furthermore, the properties of the tunnel provide a rationale for how homoserine transacetylase catalyzes a transferase reaction vs hydrolysis, despite extensive similarity in active site architecture to hydrolytic enzymes. PMID- 16313181 TI - Characterization of the 3' --> 5' exonuclease activity found in human nucleoside diphosphate kinase 1 (NDK1) and several of its homologues. AB - Nucleoside diphosphate kinases (NDKs), an evolutionarily conserved family of proteins, synthesize nucleoside triphosphates from nucleoside diphosphates and ATP. Here, we have characterized the kinase activity and DNA processing functions of eight human proteins that contain at least one domain homologous to Escherichia coli NDK. Not all human proteins with NDK-like domains exhibited NDK activity when expressed as recombinant proteins in E. coli. Human NDK1 (NM23-H1) has been reported to have 3' --> 5' exonuclease activity. In addition to human NDK1, we also find that human NDK5, NDK7, and NDK8 contain 3' --> 5' exonuclease activity. Site-directed mutagenesis, competition assays between wild-type and mutant NDK proteins, and NMR studies confirmed that the DNA-binding and 3' --> 5' exonuclease activity of human NDK1 is an intrinsic activity of the protein. Using double-stranded DNA substrates containing modified bases, human NDK1 efficiently excised nucleotides from the single-strand break produced by APE1 or Nth1. When human cells were treated with various DNA-damaging agents, human NDK1 translocated from the cytoplasm to the nucleus. These results suggest that, in addition to maintenance of nucleotide pool balance, the human NDK-like proteins may have previously unrecognized roles in DNA nucleolytic processing. PMID- 16313182 TI - Solution structure of the ligand binding domain of the fibroblast growth factor receptor: role of heparin in the activation of the receptor. AB - The three-dimensional solution structure of the ligand binding D2 domain of the fibroblast growth factor receptor (FGFR) is determined using multidimensional NMR techniques. The atomic root-mean-square distribution for the backbone atoms in the structured region is 0.64 A. Secondary structural elements in the D2 domain include 11 beta-strands arranged antiparallely into two layers of beta-sheets. The structure of the D2 domain is characterized by the presence of a short flexible helix that protrudes out of the layers of beta-sheets. Results of size exclusion chromatography and sedimentation velocity experiments show that the D2 domain exists in a monomeric state both in the presence and in the absence of bound sucrose octasulfate (SOS), a structural analogue of heparin. Comparison of the solution structure of the D2 domain with the crystal structure of the protein (D2 domain) in the FGF signaling complex reveals significant differences, suggesting that ligand (FGF) binding may induce significant conformational changes in the receptor. SOS binding sites in the D2 domain have been mapped on the basis of the 1H-15N chemical shift perturbation data. SOS binds to the positively charged residues located in beta-strand III and the flexible helix. Isothermal titration calorimetry data indicate that the ligand (hFGF-1) binds strongly (Kd approximately 10(-9) M) to the D2 domain even in the absence of SOS. Binding of SOS to either the D2 domain or hFGF-1 does not seem to be the driving force for the formation of the D2-hFGF-1 binary complex. The function of SOS binding appears to stabilize the preformed D2-FGF binary complex. PMID- 16313183 TI - Biophysical and biological studies of end-group-modified derivatives of Pep-1. AB - Pep-1 is a tryptophane-rich cell-penetrating peptide (CPP) that has been previously proposed to bind protein cargoes by hydrophobic assembly and translocate them across cellular membranes. To date, however, the molecular mechanisms responsible for cargo binding and translocation have not been clearly identified. This study was conducted to gain insight into the interaction between Pep-1 with its cargo and the biological membrane to identify the thereby involved structural elements crucial for translocation. We studied three peptides differing in their N- and C-termini: (i) Pep-1, carrying an acetylated N-terminus and a C-terminal cysteamine elongation, (ii) AcPepWAmide, with an acetylated N terminus and an amidated C-terminus, and (iii) PepW, with two free termini. Thioredoxin (TRX) and beta-galactosidase were used as protein cargoes. To study CPP-membrane interactions, we performed biophysical as well as biological assays. To mimic biological membranes, we used phospholipid liposomes in a dye leakage assay and surfactant micelles for high-resolution NMR studies. In addition, membrane integrity, cell viability, and translocation efficiency were analyzed in HeLa cells. An alpha-helical structure was found for all peptides in the hydrophobic N-terminal region encompassing residues 4-13, whereas the hydrophilic region remained unstructured in the presence of micelles. Our results show that the investigated peptides interacted with the micelles as well as with the protein cargo via their tryptophan-rich domain. All peptides displayed an orientation parallel to the micelle surface. The C-terminal cysteamine group formed an additional membrane anchor, leading to more efficient translocation properties in cells. No membrane permeabilization was observed, and our data were largely compatible with an endocytic pathway for cellular uptake. PMID- 16313184 TI - Effect of hydration on the stability of the collagen-like triple-helical structure of [4(R)-hydroxyprolyl-4(R)-hydroxyprolylglycine]10. AB - X-ray analysis has been carried out on a crystal of the collagen model peptide (Hyp(R)-Hyp(R)-Gly)10 [where Hyp(R) is 4(R)-hydroxyproline] with 1.5 A resolution. The triple-helical structure of (Hyp(R)-Hyp(R)-Gly)10 has the same helical parameters and Rich and Crick II hydrogen bond patterns as those of other collagen model peptides. However, our full-length crystal structure revealed that almost all consecutive Hyp(R) residues take the up-up pucker in contrast to putative down-up puckering propensities of other collagen model peptides. The unique feature of thermodynamic parameters associated with the conformational transition of this peptide from triple helix to single coil is that both enthalpy and entropy changes of the transition are much smaller than those of other model peptides such as (Pro-Pro-Gly)10 and (Pro-Hyp(R)-Gly)10. To corroborate the precise structural information including main- and side-chain dihedral angles and intra- and interwater bridge networks, we estimated the degrees of hydration by comparing molecular volumes observed experimentally in solution to those calculated ones from the crystal structure. The results showed that the degree of hydration of (Hyp(R)-Hyp(R)-Gly)10 is comparable to that of (Pro-Hyp(R)-Gly)10 in the triple-helical state, but the former was more highly hydrated than (Pro Hyp(R)-Gly)10 in the single-coil state. Because hydration reduces the enthalpy due to the formation of a hydrogen bond with a water molecule and diminishes the entropy due to the restriction of water molecules surrounding a peptide molecule, we concluded that the high thermal stability of (Hyp(R)-Hyp(R)-Gly)10 is able to be described by its high hydration in the single-coil state. PMID- 16313185 TI - Single giant unilamellar vesicle method reveals effect of antimicrobial peptide magainin 2 on membrane permeability. AB - It is thought that magainin 2, an antimicrobial peptide, acts by binding to lipid membranes. Recent studies using a suspension of large unilamellar vesicles (LUVs) indicate that magainin 2 causes gradual leakage from LUVs containing negatively charged lipids. However, the details of the characteristics of the membrane permeability and the mechanism of pore formation remain unclear. In this report, we investigated the interaction of magainin 2 with single giant unilamellar vesicles (GUVs) composed of a dioleoylphosphatidylcholine and dioleoylphosphatidylglycerol mixture (50% DOPG/50% DOPC GUVs) containing the fluorescent dye, calcein, by phase contrast, fluorescence microscopy using the single GUV method. Low concentrations (3-10 microM) of magainin 2 caused the rapid leakage of calcein from single GUVs but did not disrupt the liposomes or change the membrane structure, showing directly that magainin 2 forms membrane pores through which calcein leaked. The rapid leakage of calcein from a GUV started stochastically, and once it began, the complete leakage occurred rapidly (6-60 s). The fraction of completely leaked GUV, P(L), increased with time and also with an increase in magainin 2 concentration. Shape changes in these GUVs occurred prior to the pore formation and also at lower concentrations of magainin 2, which could not induce the pore formation. Their analysis indicates that binding of magainin 2 to the external monolayer of the GUV increases its membrane area, thereby raising its surface pressure. The addition of lysophosphatidylcholine into the external monolayer of GUVs increased P(L). On the basis of these results, we propose the two-state transition model for the pore formation. PMID- 16313186 TI - Nuclear magnetic resonance structural studies of a potassium channel charybdotoxin complex. AB - Ion channels play critical roles in signaling processes and are attractive targets for treating various diseases. Here we describe an NMR-based strategy for structural analyses of potassium channel-ligand complexes using KcsA (residues 1 132, with six mutations to impart toxin binding and to mimic the eukaryotic hERG channel). Using this approach, we determined the solution structure of KcsA in complex with the high-affinity peptide antagonist charybdotoxin. The structural data reveal how charybdotoxin binds to the closed form of KcsA and makes specific contacts with the extracellular surface of the ion channel, resulting in pore blockage. This represents the first direct structural information about an ion channel complexed to a peptide antagonist and provides an experimental framework for understanding and interpreting earlier mutational analyses. The strategy presented here overcomes many of the limitations of conventional NMR approaches to helical membrane protein structure determination and can be applied in the study of the binding of druglike molecules to this important class of proteins. PMID- 16313187 TI - The extracellular domains of ErbB3 retain high ligand binding affinity at endosome pH and in the locked conformation. AB - The extracellular, ligand binding regions of ErbB receptors consist of four domains that can assume at least two alternative conformations, extended and locked. The locked conformation, observed in several crystal structures, is held together by a noncovalent intramolecular tether and is incompatible with current models for receptor dimerization and ligand activation. Based on structures of ligand-receptor complexes in the extended conformation, the high affinity ligand binding pocket between domains I and III is disrupted in the locked conformation. Therefore the biological role of the locked conformation is not clear. To address the impact of the locked conformation on ligand binding, we compared extracellular domains of wild-type ErbB3, mutant domains in a constitutively locked or extended conformation and partial extracellular domain constructs. We found that the constitutively locked receptor domains and truncated constructs carrying only domains I-II or III-IV strongly bind ligand, albeit with reduced affinity compared to wild-type receptor. This suggests that the locked conformation cannot be discounted for ligand binding. The significant binding by both partial interfaces in domains I and III also suggests that "partial bivalency" may be the reason for the low nanomolar and high picomolar binding observed for ErbB3 in the respective "low" and high affinity states. In contrast to EGFR (ErbB1), ErbB3 retains high ligand binding affinity at an endosome comparable pH in both the extended and locked conformations. Ligand affinity for the locked conformation even improves at low pH. For ErbB3, the contribution of domain I to ligand binding is strong and increases at low pH while its contribution is thought to be minimal for EGFR, regardless of pH. This shift in domain contribution and pH dependency provides a mechanistic explanation for some of the divergent properties of EGFR and ErbB3. PMID- 16313188 TI - Detection of human betaV-tubulin expression in epithelial cancer cell lines by tubulin proteomics. AB - Tubulin, the constitutive protein of microtubules, is a heterodimeric protein with an alpha and beta subunit, encoded in vertebrates by six and seven different genes, respectively. Each tubulin isotype can be identified by its divergent C terminal sequence. Nevertheless, two groups of beta-tubulin isotypes can be distinguished by sequence alignment; one includes betaI-, betaII-, betaIVa-, and betaIVb-tubulin, and the other includes betaIII-, betaV-, and betaVI-tubulin. betaIII-tubulin overexpression has been associated with microtubule destabilization and resistance to Taxol. Recent data indicate that mouse betaV tubulin overexpression in CHO cells results in profound microtubule disorganization and dependence of cells on Taxol for growth. Mouse and human betaV-tubulin sequences display several differences, such as their respective extreme C-terminus, suggesting that they may have different effects on microtubule stability and different affinities for drugs. When high-resolution isoelectric focusing, in-gel CNBr cleavage, and mass spectrometry were combined, we detected for the first time the betaV-tubulin protein in human cell lines and found that it was highly expressed in Hey, an epithelial ovarian cancer cell line. Our data confirm that human and rodent betaV-tubulins are distinct and indicate that, regardless of species, betaIII- and betaV-tubulin may be expressed in a complementary pattern at the protein level. Therefore, both betaIII- and betaV-tubulin expression levels should be systematically determined to assess the role of differential tubulin isotype expression in the response of tumors to drugs targeting microtubules. PMID- 16313189 TI - Structure-based identification of the binding site for the hemiasterlin analogue HTI-286 on tubulin. AB - A binding mode of HTI-286, a synthetic analogue of the peptidic antimitotic agent hemiasterlin, to tubulin is proposed. The binding mode was derived from iterative docking experiments directed at regions of the tubulin interdimer interface that are believed to be consistent with all current experimental data regarding the HTI-286/tubulin interaction. These data include (1) competitive inhibition of the tubulin binding of the Vinca alkaloids and other antimitotic agents, (2) proximity to stretches of amino acid residues identified in two separate photoaffinity-labeling experiments, (3) structure-activity relationships for HTI 286 and its analogues, (4) saturation transfer difference nuclear magnetic resonance (NMR) experiments, and (5) NMR transfer nuclear Overhauser effect spectroscopy (NOESY) experiments that potentially identify the bioactive conformation. The predicted binding mode thus affords a means to understand the mode of action of hemiasterlin, HTI-286, and other closely related molecules. PMID- 16313190 TI - Polymorphism at residue 129 modulates the conformational conversion of the D178N variant of human prion protein 90-231. AB - One of the arguments in favor of the protein-only hypothesis of transmissible spongiform encephalopathies is the link between inherited prion diseases and specific mutations in the PRNP gene. One such mutation (Asp178 --> Asn) is associated with two distinct disorders: fatal familial insomnia or familial Creutzfeldt-Jakob disease, depending upon the presence of Met or Val at position 129, respectively. In this study, we have characterized the biophysical properties of recombinant human prion proteins (huPrP90-231) corresponding to the polymorphic variants D178N/M129 and D178N/V129. In comparison to the wild-type protein, both polymorphic forms of D178N huPrP show a greatly increased propensity for a conversion to beta-sheet-rich oligomers (at acidic pH) and thioflavine T-positive amyloid fibrils (at neutral pH). Importantly, the conversion propensity for the D178N variant is strongly dependent upon the M/V polymorphism at position 129, whereas under identical experimental conditions, no such dependence is observed for the wild-type protein. Amyloid fibrils formed by wild-type huPrP90-231 and the D178N variant are characterized by different secondary structures, and these structures are further modulated by residue 129 polymorphism. Although on the basis of only in vitro data, this study strongly suggests that polymorphism-dependent phenotypic variability of familial prion diseases may be linked to differences in biophysical properties of prion protein variants. PMID- 16313191 TI - The activity of the lactose transporter from Streptococcus thermophilus is increased by phosphorylated IIA and the action of beta-galactosidase. AB - The metabolism of lactose by Streptococcus thermophilus is highly regulated, allowing the bacterium to prefer lactose over glucose as main source of carbon and energy. In vitro analysis of the enzymes involved in transport and hydrolysis of lactose showed that the transport reaction benefits from the hydrolysis of lactose at the trans side of the membrane. Furthermore, the activity of LacS is modulated by PEP-dependent phosphorylation of the IIA domain via the general energy coupling proteins of the PTS, Enzyme I and HPr. To determine whether unphosphorylated LacS-IIA inhibited, or the phosphorylated form stimulated lactose counterflow, a LacS-IIA truncation mutant of LacS was constructed. Detailed analyses of transport in whole cells and in proteoliposomes indicated that unphosphorylated LacS-IIA does not functionally interact with the carrier domain. Instead, interaction of the phosphorylated form of LacS-IIA with the carrier stimulates lactose counterflow transport. The proposed mode of regulation thus proceeds via a mechanism opposite to the inducer exclusion type of regulation in gram-negative bacteria, where transporters are inhibited by binding of the unphosphorylated form of IIA(Glc). PMID- 16313192 TI - Conformational studies of the tetramerization site of human erythroid spectrin by cysteine-scanning spin-labeling EPR methods. AB - We used cysteine-scanning and spin-labeling methods to prepare singly spin labeled recombinant peptides for electron paramagnetic resonance studies of the partial domain regions at the tetramerization site (N-terminal end of alpha and C terminal end of beta) of erythroid spectrin. The values of the inverse line width parameter (deltaH0(-1)) from a family of Sp alphaI-1-368delta peptides scanning residues 21-30 exhibited a periodicity of approximately 3.5-4. We used molecular dynamics calculations to show that the asymmetric mobility of this helix is not necessarily due to tertiary contacts, but is likely due to intrinsic properties of helix C', a helix with a heptad pattern sequence. The residues with low deltaH0(-1) values (residues at positions 21, 25, and 28/29) were those on the hydrophobic side of this amphipathic helix. Native gel electrophoresis results showed that these residues were functionally important and are involved in the tetramerization process. Thus, EPR results readily identified functionally important residues in the alpha spectrin partial domain region. Mutations at these positions may lead to clinical symptoms. Similarly, the deltaH0(-1) values from a family of spin-labeled Sp betaI-1898-2083delta peptides also exhibited a periodicity of approximately 3.5-4, indicating a helical conformation in the two scanned regions (residues 2008-2018 and residues 2060-2070). However, the region consisting of residues 2071-2076 was in a disordered conformation. Both helical regions include a hydrophilic side with high deltaH0(-1) values and a hydrophobic side with low deltaH0(-1) values, demonstrating the amphipathic nature of the helical regions. Residues 2008, 2011, 2014, and 2018 in the first scanned region and residues 2061, 2065, and 2068 in the second scanned region were on the hydrophobic side. These residues were critical in alphabeta spectrin association at the tetramerization site. Mutations at some of these positions have been reported to be detrimental in clinical studies. PMID- 16313193 TI - Peripheral stator of the yeast V-ATPase: stoichiometry and specificity of interaction between the EG complex and subunits C and H. AB - V-ATPases are multisubunit membrane protein complexes that use the energy provided by ATP hydrolysis to generate a proton gradient across various intracellular and plasma membranes. In doing so, they maintain an acidic pH in the lumen of intracellular organelles and acidify extracellular milieu to support specific cellular functions. V-ATPases are structurally similar to the F1F0-ATP synthase, with an intrinsic membrane domain (V0) and an extrinsic peripheral domain (V1) joined by several connecting elements. To gain a clear functional understanding of the catalytic mechanism, and of the stability requirements for regulatory processes in the enzyme, a clear topology of the enzyme has to be established. In particular, the composition and arrangement of the peripheral stator subunits must be firmly settled, as these play specific roles in catalysis and regulation. We have designed a strategy allowing us to coexpress different combinations of these subunits to delineate specific interactions. In this study, we report the interaction between the peripheral stator EG complex and subunits C and H of the V-ATPase from the yeast Saccharomyces cerevisae. A combination of analytical gel filtration, native gel electrophoresis, and ultracentrifugation analysis allowed us to ascertain the homogeneity and molar mass of the purified EGC complex as well as of the EG complex, supporting the formation of 1:1(:1) stoichiometric complexes. The EGC complex can be formed in vitro by combining equimolar amounts of subunit C and the EG subcomplex and results most likely from the initial interaction between subunits E and C. PMID- 16313194 TI - Coating proteins: structure and cross-linking in fp-1 from the green shell mussel Perna canaliculus. AB - The protein family known as fp-1 provides mussel byssus with a protective outer coating and has drawn much attention for its water resistant bioadhesive properties in vitro. A new fp-l isolated from the green shell mussel Perna canaliculus (pcfp-1) reveals a composition dominated by only four amino acids: 3,4-dihydroxyphenyl-L-alanine (dopa), lysine, proline, and valine at approximately 20 mol % each. SDS-PAGE and MALDI-TOF mass spectrometry detected size variants at 48 and 52 kDa in preparations of purified Pcfp-1. The N-terminal sequence enabled construction of oligonucleotide primers for PCR and RACE-derived cDNAs from which the complete sequence of four variants was deduced. pcfp-1 deviates from all known homologues in other mussels in several notable respects: its mass is half, most of its sequence is represented by 75 tandem repeats of a tetrapeptide, i.e., PY*VK, in which Y* is dopa, prolines are not hydroxylated, and thiolate cysteines are clustered in homologous sequences at both the amino and carboxy termini. Amino acids in the repeat sequence show a striking resemblance to proline-rich cell wall proteins with tandemly repeated PPVYK pentapeptides [Hong, J. C., Nagao, R. T., and Key, J. L. (1987) J. Biol. Chem. 262, 8367-8376]. Cysteine plays a key role in cross-linking pcfp-1 by forming adducts with dopaquinone. Significant 5-S-cysteinyldopa and smaller amounts of 2 S-cysteinyldopa were detected in hydrolysates of the byssal threads of P. canaliculus. The cross-links could also be formed by oxidation of pcfp-1 in vitro using mushroom tyrosinase. Cysteinyldopa cross-links were present in trace amounts only in the byssus of other mussel species. PMID- 16313195 TI - 1H NMR and optical spectroscopic investigation of the sequence-dependent dimerization of a symmetrical cyanine dye in the DNA minor groove. AB - A symmetrical cyanine dye was previously shown to bind as a cofacial dimer to alternating A-T sequences of duplex DNA. Indirect evidence suggested that dimerization of the dye occurred in the minor groove. 1H NMR experiments reported here verify this model based on broadening and shifting of signals due to protons on carbon 2 of adenine and imino protons at the central five A-T pairs of the 11 base pair duplex: 5'-GCGTATATGCG-3'/3'-CGCATATACGC-5'. This binding mode is similar to that of distamycin A, even though the dye lacks the hydrogen-bonding groups used by distamycin for sequence-specific recognition. Surprisingly, the third base pair (G-C) was also implicated in the binding site. UV-vis experiments were used to compare the extent of dimerization of the dye for 11 different sequence variants. These experiments verified the importance of a G-C pair at the third position: replacing this pair with A-T suppressed dimerization. These results indicate that the dye binding site spans six base pairs: 5'-GTATAT-3'. The initial G-C pair seems to be important for widening the minor groove rather than for making important contacts with the dye molecules since inverting its orientation to C-G or replacing it with I-C still led to favorable dimerization of the dye. PMID- 16313196 TI - Reaction of serine-glyoxylate aminotransferase with the alternative substrate ketomalonate indicates rate-limiting protonation of a quinonoid intermediate. AB - Serine-glyoxylate aminotransferase (SGAT) from Hyphomicrobium methylovorum is a pyridoxal 5'-phosphate (PLP) enzyme that catalyzes the interconversion of L serine and glyoxylate to hydroxypyruvate and glycine. The primary deuterium isotope effect using L-serine 2-D is one on (V/K)serine and V in the steady state. Pre-steady-state experiments also indicate that there is no primary deuterium isotope effect with L-serine 2-D. The results suggest there is no rate limitation by abstraction of the alpha proton of L-serine in the SGAT reaction. In the steady-state a solvent deuterium isotope effect of about 2 was measured on (V/K)L-serine and (V/K)ketomalonate and about 5.5 on V. Similar solvent isotope effects were observed in the pre-steady-state for the natural substrates and the alternative substrate ketomalonate. In the pre-steady-state, no reaction intermediates typical of PLP enzymes were observed with the substrates L-serine, glyoxylate, and hydroxypyruvate. The data suggest that breakdown and formation of the ketimine intermediate is the primary rate-limiting step with the natural substrates. In contrast, using the alternative substrate ketomalonate, pre-steady state experiments display the transient formation of a 490 nm absorbing species typical of a quinonoid intermediate. The solvent isotope effect results also suggest that with ketomalonate as substrate protonation at C(alpha) is the slowest step in the SGAT reaction. This is the first report of a rate-limiting protonation of a quinonoid at C(alpha) of the external Schiff base in an aminotransferase reaction. PMID- 16313197 TI - Mechanism of A beta(1-40) fibril-induced fluorescence of (trans,trans)-1-bromo 2,5-bis(4-hydroxystyryl)benzene (K114). AB - K114, (trans,trans)-1-bromo-2,5-bis(4-hydroxystyryl)benzene, is a fluorescent Congo Red analogue that binds tightly to amyloid fibrils, but not the monomeric proteins, with a concomitant enhancement in fluorescence. The mechanism for the low aqueous fluorescence and the subsequent enhancement by A beta(1-40) fibrils was investigated by fluorescence spectroscopy and binding analysis. K114's unusually low buffer fluorescence is due to self-quenching in sedimentable aggregates or micelles which upon interacting with amyloid fibrils undergo an enhancement in fluorescence intensity and shifts in the excitation and emission spectra. These spectral changes are suggestive of a stabilization of the phenolate anion, perhaps by hydrogen bonding, rather than an increase in the microenvironment dielectric constant or dye immobilization. 1,4-Bis(4 aminophenylethenyl)-2-methoxybenzene, which lacks the phenol moiety, and X-34, which contains a stabilized phenol (pK approximately 13.4), do not display the phenolate anion fluorescence in the presence of fibrils. The apparent affinity of K114 for fibril binding is 20-30 nM with a stoichiometry of 2.2 mol of K114/mol of A beta(1-40) monomer. Competition studies indicate that K114 and Congo Red share a site, but K114 does not bind to sites on A beta(1-40) fibrils for neutral benzothiazole (BTA-1), cationic thioflavin T, or the hydrophobic (S)-naproxen and (R)-ibuprofen molecules. Comparison of benzothiazole binding stoichiometry which has been suggested to reflect disease-relevant amyloid structures to that of Congo Red analogues which reflect total fibril content may be useful in defining biologically pertinent conformational forms of amyloid. PMID- 16313198 TI - Novel aromatic ester from Piper longum and its analogues inhibit expression of cell adhesion molecules on endothelial cells. AB - We report here the isolation and characterization of two active principles, ethyl 3',4',5'-trimethoxycinnamate (1) and piperine (2), from the combined hexane and chloroform extracts of Piper longum. Using primary human umbilical vein endothelial cells, we evaluated the activities of compound 1 on TNF-alpha-induced expression of cell adhesion molecules, viz., ICAM-1, VCAM-1, and E-selectin, which play key roles in controlling various inflammatory diseases. Both compounds 1 and 2 inhibited the TNF-alpha-induced expression of ICAM-1 in a dose- and time dependent manner; however, the activity of ethyl 3',4',5'-trimethoxycinnamate (1) was approximately 1.3 times higher than that of piperine (2). As ethyl 3',4',5' trimethoxycinnamate (1) has been isolated for the first time from a natural source, Piper longum, and it exhibited higher activity, we carried out further studies on it. To correlate its cell adhesion molecule inhibitory activity with its functional consequences, we showed that it significantly blocked the adhesion of neutrophils to endothelium in a time- and concentration-dependent manner. Importantly, the inhibitory effect of cinnamate 1 was found to be reversible. To elucidate its structure-function-activity relationship, we synthesized nine different analogues of ethyl 3',4',5'-trimethoxycinnamate, i.e., compounds 3-11, and compared the ICAM-1 inhibitory activity of compound 1 with those of its synthetic analogues as well as the corresponding acids 12-15. The structure activity studies indicate that the chain length of the alcohol moiety, substituents in the aromatic ring, and alpha, beta-double bond of the cinnamic acid ester have significant effects on the inhibition of TNF-alpha-induced expression of ICAM-1 on endothelial cells. These findings have implications in developing compounds with a better therapeutic index against various inflammatory diseases. PMID- 16313199 TI - Optical spectra of lactoperoxidase as a function of solvent. AB - The iron of lactoperoxidase is predominantly high-spin at ambient temperature. Optical spectra of lactoperoxidase indicate that the iron changes from high-spin to low-spin in the temperature range from room temperature to 20 K. The transformation is independent of whether the enzyme is in glycerol/water or solid sugar glass. Addition of the inhibitor benzohydroxamic acid increases the amount of the low-spin form, and again the transformation is independent of whether the protein is in an aqueous solution or a nearly anhydrous sugar. In contrast to lactoperoxidase, horseradish peroxidase remains high-spin over the temperature excursion in both solvents and with addition of benzohydroxamic acid. We conclude that details of the heme pocket of lactoperoxidase allow ligation changes with temperature that are dependent upon the apoprotein but independent of solvent fluctuations. At low pH, lactoperoxidase shows a solvent-dependent transition; the high-spin form is predominant in anhydrous sugar glass, but in the presence of water, the low-spin form is also present in abundance. The active site of lactoperoxidase is not as tightly constrained at low pH as at neutrality, though the enzyme is active over a wide pH range. PMID- 16313201 TI - Ultraviolet absorption spectra of substituted phenols: a computational study. AB - Vertical excitation energies for electronic transitions from the ground state to the first two excited states of phenol, mono- and disubstituted methoxyphenols and methyl-substituted phenols have been characterized with the Time-Dependent Density Functional Theory (TD-DFT), the Complete Active Space Self-Consistent Field method (CASSCF) and the Coupled Cluster with Single and Double Excitations Equation-of-Motion approach (CCSD-EOM) to simulate and interpret experimental ultraviolet absorption spectra. While CASSCF excitation energies for the first two transitions either are grossly overestimated or exhibit a weak correlation with experimental data, both TD-DFT and CCSD-EOM perform very well, reproducing the spectral shifts of both the primary band and secondary band observed upon substitution. The conformational dependence of the calculated excitation energies is generally smaller than the shifts caused by substitution. PMID- 16313202 TI - A compulsory experiential and inter-professional rural health subject for undergraduate students. AB - CONTEXT: The Australian University Departments of Rural Health (UDRH) program aims to improve the rural workforce through improving rural experiences of undergraduates, graduates and health professionals. The 10 UDRHs vary in the way they implement the UDRH Program. This article describes the development of a compulsory, experiential and inter-professional rural health subject in a medical curriculum by the UDRH at the University of Melbourne, Victoria, Australia. The key concepts, underpinning theory, processes, logistics and resources required to develop and implement the subject among a network of small towns and Indigenous communities are examined. METHODS: METHODS used included formative evaluation using a participatory research approach, including the examination of internal discussion papers, minutes of meetings and planning workshops, financial records and feedback from students, staff and preceptors. ISSUES: Students, staff and preceptors generally accept the value of the rural health module in professional training. However, they stress the need for explicit learning objectives, relevant content and engaging delivery. Students expressed some concern about the compulsory nature of the subject. Significant issues are capacity, quality, sustainability and managing expectations of government and communities. Capacity includes the availability of appropriate placements and preceptors who have the capacity and capability to teach or to be supported to teach. The availability of appropriate and affordable accommodation is important, as are the cost and safety of travel to and from placements. Sustainability of 'placements' and 'placement fatigue' must be managed with a combination of resources and preceptor training and support. Quality of placements is important and highlights a paradox in that, while academically rigorous teaching, research and clinical programs for students and preceptors are important for quality, they are mostly perceived as additional burdens by health service providers. Finally, the expectations and commitment of stakeholders, in particular governments and communities, for student placements must be realistic and backed with resources. LESSONS LEARNED: A compulsory experiential inter-professional subject for undergraduate students is possible but requires significant resources and commitment over the long term. The resources must include funding for: preceptor time, training and support; recruitment and retention of appropriately qualified academic clinicians; intersectoral strategies like improved public transport and safe roads; community and other stakeholder engagement to develop and maintain educational facilities. Last but not least, funding is essential to support the long-term evaluation and monitoring of the outcomes in terms of the quality and quantity of the rural workforce over time. PMID- 16313205 TI - Starting early: promoting the mental health of women and girls throughout the life span. AB - The importance of mental health in the promotion of lifelong health among men and women alike cannot be overstated. However, mental health remains under-addressed within general public health and community health programs. In this report, we focus primarily on the mental health of women and discuss risk factors that can affect the well-being of women throughout the life span. The literature reviewed demonstrates a strong relationship between social and environmental risk factors, such as abuse and family dysfunction in childhood, to health risk behaviors and poor mental health in adulthood. We concluded that adverse childhood experiences (ACEs) and poor adult mental health could contribute to cycles of intergenerational transmission of risks leading to poor mental and physical health in children of ACEexposed parents. Also, we argue that public health communities can make a difference in women's lifelong health by improving early recognition and treatment of mental health concerns, seeking opportunities to prevent exposures to known risk factors in childhood, and developing targeted parenting interventions. Promoting healthy psychological states and coping mechanisms before, during, and after exposure to adverse events throughout life is also critical. Perhaps such efforts will help to reduce or even break cycles of risk exposure specifically for women and their children. Finally, existing prevention activities and opportunities for promoting the mental health of girls and women are discussed. Ultimately, this report challenges the women's health and public health communities to take action because mental health can have a serious impact on lifelong well-being. PMID- 16313206 TI - Biomedical research on health and performance of military women: accomplishments of the Defense Women's Health Research Program (DWHRP). AB - In 1994, Congress provided dollar 40 M for biomedical research on issues of importance for military women. This supported 104 intramural and 30 extramural studies and launched an era of research to narrow the knowledge gap on protection and enhancement of health and performance of military women. Projects addressed issues specific to female physiology (e.g., gynecological health in the field, maternal malaria), problems with higher prevalence for women (e.g., marginal iron deficiency, stress fracture), and issues of drug and materiel safety that had only been extrapolated from studies of men (e.g., chemical agent prophylaxis, fatigue countermeasures). Several important assumptions about female physiology and occupational risks were found to be astoundingly wrong. Hormonal changes through the menstrual cycle were less important to acute health risks and performance than predicted, exercise did not increase risk for amenorrhea and consequent bone mineral loss, and women tolerated G-forces and could be as safe as men in the cockpit if their equipment was designed for normal size and strength ranges. Data on personal readiness issues, such as body fat, physical fitness, nutrition, and postpartum return to duty, allowed reconsideration of standards that were gender appropriate and not simply disconnected adjustments to existing male standards. Other discoveries directly benefited men as well as women, including development of medical surveillance databases, identification of task strength demands jeopardizing safety and performance, and greater understanding of the effects of psychosocial stress on health and performance. This surge of research has translated into advances for the welfare of service women and the readiness of the entire force; relevant gender issues are now routine considerations for researchers and equipment developers, and some key remaining research gaps of special importance to military women continue to be investigated. PMID- 16313207 TI - Toward optimal health: Donna E. Stewart, M.D. discusses perinatal depression. Interview by Jodi R. Godfrey. PMID- 16313208 TI - Menstrual and reproductive factors and fracture risk: the Leisure World Cohort Study. AB - BACKGROUND: Because menopausal estrogen is related to the development of osteoporosis, we investigated the potential associations of the estrogen-related events of menarche, pregnancy, and menopause with fracture risk in a population based, prospective study of older women. METHODS: The Leisure World Cohort was established in the early 1980s when residents of a California retirement community, including 8877 women, completed a health survey. Incident fractures of the hip (n = 1220), wrist (n = 463), and spine (n = 613) incurred over two decades were identified from four follow-up questionnaires, hospital discharge records, and (for hip fracture) death certificates. Hazard ratios (HR) adjusted for age and other potential confounders were calculated using proportional hazards regression. RESULTS: Late age at menarche was associated with decreased hip fracture risk (HR = 0.84, 95% CI 0.72-0.98, for age > or = 14 vs. < or = 12 years) but was unrelated to fractures at other sites. Hip fracture risk was also reduced in women who had been pregnant (HR = 0.83, 95% CI 0.72- 0.95). Women who reported menopause at age 45+ had a lower risk of wrist fracture compared with those with menopause at age < or = 44 (HR = 0.74, 95% CI 0.58-0.95 for ages 45 54; HR = 0.71, 95% CI 0.49-1.04 for ages 55+). Although fracture risks did not differ between ever and never users of menopausal estrogen, recency since last use was related to wrist fractures (HR = 1.09, 95% CI 1.03-1.16 for each 5 years since last years). CONCLUSIONS: The estrogen-related events of menarche, pregnancy, and menopause were not associated with osteoporotic fracture risk in a consistent manner. Other factors related to these events may be influencing development of osteoporosis and the likelihood of sustaining a fracture in older women. PMID- 16313209 TI - Does sex matter in the associations between classic risk factors and fatal coronary heart disease in populations from the Asia-Pacific region? AB - BACKGROUND: There is much interest in promoting healthy heart awareness among women. However, little is known about the reasons behind the lower rates of heart disease among women compared with men, and why this risk difference diminishes with age. Previous comparative studies have generally had insufficient numbers of women to quantify such differences reliably. METHODS: We carried out an individual participant data meta-analysis of 39 cohort studies (32 from Asian countries and 7 from Australia and New Zealand). Cox models were used to estimate hazard ratios (HR) for coronary death, comparing men to women. Further adjustments were made for several proven coronary risk factors to quantify their contributions to the sex differential. Sex interactions were tested for the same risk factors. RESULTS: During 4 million person-years of follow-up, there were 1989 (926 female) deaths from coronary heart disease (CHD). The age-adjusted and study-adjusted male/female HR (95% confidence interval [95% CI]) was 2.05 (1.89 2.22). At baseline, 54% of men vs. 7% of women were current smokers; hence, adjustment for smoking explained the largest component (20%) of this HR. A significant sex interaction was observed between systolic blood pressure (SBP) and CHD mortality such that a 10 mm Hg increase was associated with a 15% greater increase in the relative risk (RR) of coronary death in women compared with men (p = 0.002). CONCLUSIONS: Only a small amount of the sex differential in coronary death could be explained by differences in the prevalence of classic risk factors. Alternative explanations are required to explain the age-related attenuation of the sex difference in CHD risk. PMID- 16313210 TI - Intimate partner violence and disabilities among women attending family practice clinics. AB - PURPOSE: To estimate the frequency and type of disabilities preventing work among those experiencing intimate partner violence (IPV) compared with those never experiencing IPV. METHODS: We used a large cross-sectional survey of women, ages 18-65, attending family practice clinics from 1997 through 1998. Participation included a 5-10-minute in-clinic survey assessing IPV experience and a longer telephone survey assessing health status and chronic disabilities that prevented work outside the home or housework. RESULTS: Of 1,152 eligible women surveyed, 54% experienced some type of IPV, and 24% were currently in a violent relationship. Women who had ever experienced IPV were more than twice as likely to report a disability (adjusted odds ratio [aOR] = 2.2, 95% confidence interval [CI] 1.6, 3.0). The most commonly reported disabilities were those associated with heart or circulatory disease (4.9%), followed by back problems (3.5%), chronic pain (3.4%), arthritis (3.0%), nerve system damage (2.4%), asthma or another respiratory problem including emphysema (1.7%), and either depression (1.6%) or another mental illness (1.0%). Women ever experiencing IPV were more likely to report a disability due to generalized chronic pain (aOR = 2.5, 95% CI 1.5, 4.3) and mental illness (aOR = 4.5, 95% CI 1.5, 13.1). IPV-related injuries were associated in a dose-dependent manner with having any disability and with disability from chronic pain, asthma and other respiratory diseases, mental illness, and chronic diseases. CONCLUSIONS: Primary care-based efforts to screen for IPV and effectively intervene to reduce the impact of IPV on women's lives must be a public health priority to reduce the shortterm and long-term health effects, including disabilities. PMID- 16313211 TI - Surgical management of complex obstetric fistula in Eritrea. AB - OBJECTIVES: To evaluate the incidence of and demographic characteristics associated with obstetric fistula in Eritrea. To determine the outcomes of surgical repair of complex fistula in Eritrea by a visiting surgical team. METHODS: A surgical team comprising expert gynecologic surgeons traveled to Eritrea in September 2004. We evaluated 50 patients with genitourinary fistula and performed surgical repairs of these fistulas on 37 women via both vaginal and abdominal approaches. Demographic and basic medical data were obtained at the time of evaluation, and follow-up questionnaires were completed at 4 weeks postoperative. RESULTS: The majority of the women had fistulas related to obstructed labor at their first pregnancy unattended by any healthcare professional. The average duration of labor was 3 days, and more than half had resulted in stillbirths. The rate of successful repair in women with primary vesicovaginal fistulas (VVF) was 63%, and that in women with recurrent vesicovaginal fistulas was 61%. Two women required urinary diversion procedures because of the severity of the damage to the genital tract. Urethral reconstruction in women with urethrovaginal fistulas (UVFs) was successfully accomplished in 77% of patients. The rate of successful repair of rectovaginal fistulas (RVFs) was 87%. CONCLUSIONS: We have demonstrated that a team of specialized surgeons can successfully accomplish surgical procedures and repairs of very complex urinary tract fistulas in a very short mission to a resource-poor nation. PMID- 16313212 TI - Perceived risk of breast cancer among women at average and increased risk. AB - BACKGROUND: The accuracy of a woman's perception of her risk of developing breast cancer has gained importance as more options for primary prevention have become available for those at increased risk. Conversely, women at average risk who perceive themselves as at increased risk may suffer from avoidable anxiety or unnecessary treatment. This study examined characteristics associated with perception of breast cancer risk among women at average and increased risk. METHODS: We included 1700 women 40-74 years old without a history of breast cancer. The outcome variable was a woman's perceived lifetime risk of developing breast cancer. The Gail model was used to categorize a woman's actual risk as average or high. Multivariate logistic regression models were used to model a woman's perception that her risk was (1) higher than average for those whose Gail score indicated average risk (<1.67% 5-year risk) and (2) accurate for those whose Gail score indicated increased risk (> or = 1.67% 5-year risk). RESULTS: Of women at average risk, 72%, but only 43% of those at high risk, accurately perceived their risk. Among women at average risk, those who were younger, had a family history of breast cancer, had no history of childbirth, or had more frequent exposure to lay media information about breast health were more likely than women without these characteristics to overestimate their future risk. Among women at increased risk, younger women and those with a family history of breast cancer were more likely than women without these characteristics to accurately perceive their increased risk. African American women were less likely than white women to accurately perceive their risk. CONCLUSIONS: A majority of women at high risk of developing breast cancer underestimate their risk, and a substantial proportion of women at average risk perceive they are at increased risk. PMID- 16313213 TI - Educational attainment but not literacy is associated with HIV risk behavior among incarcerated women. AB - PURPOSE: To identify the educational factors associated with HIV risk behaviors among incarcerated women. METHODS: We evaluated a cohort of female detainees at The Rhode Island Adult Correctional Institute between July and September 2004. Among the 423 women who participated in the study, 55% did not have a high school diploma, 29% had < or = 8th grade reading capacity, 32% had a learning disability, 37% had problem drinking, and 61% (257/423) reported HIV risk behavior. RESULTS: In multiple logistic regression, participants who had completed high school had the lowest likelihood of HIV risk behavior (adjusted odds ratio [OR] 0.35, 95% confidence interval [CI] 0.12-1.00). There was no association between participants? literacy level and HIV risk behavior (adjusted OR 2.02, 95% CI 0.83-4.92). CONCLUSIONS: Correctional education programs to reduce HIV risk behavior should focus on those with low educational attainment irrespective of literacy skills. PMID- 16313214 TI - FDA's decision on Plan B: a surrogate for lack of commitment to women's health. PMID- 16313215 TI - Women in the military: new perspectives, new science. PMID- 16313216 TI - Hard questions concerning fistula surgery in Third World countries. PMID- 16313217 TI - Shared medical appointments: facilitating interdisciplinary care for midlife women. AB - Shared medical appointments (SMAs) are a new way to deliver woman-focused interdisciplinary care for midlife women. SMAs are a series of one physician to one patient encounters with other patient observers. The women's health physician addresses each woman's unique medical needs individually but in the context of a shared setting. The major focus is on delivering individual medical care with the benefits of additional time spent educating women patients and answering questions. PMID- 16313219 TI - The SENS challenge: 20,000 US dollars says the foreseeable defeat of aging is not laughable. PMID- 16313220 TI - Two hands make light work of gene modification. AB - The full power of gene modification in higher eukaryotes, including its potential to correct disease-causing mutations, has so far been limited by its low efficiency. Now that power looks set to be released through the use of customized endonuclease "hands," DNA-binding zinc fingers linked to nuclease "thumbs," capable of grasping and cutting any chosen genomic target locus. Once cleaved, the target locus is efficiently repaired by endogenous mechanisms, using an exogenous DNA template that carries the chosen modification. These helping hands are likely to touch many areas of biological and clinical endeavor. PMID- 16313221 TI - Mitochondrial DNA mutations, apoptosis, and the misfolded protein response. AB - Studies of transgenic mice with accelerated accumulation of mtDNA mutations specifically in the heart lead us to propose that apoptotic signaling and cell death is central to the pathogenesis of mtDNA mutations in aging. It is the cellular response to that apoptotic signaling and the organ?s compensatory response to a loss of cells that specify the phenotype of an accumulation of mtDNA mutations. In the heart, cardiomyocytes induce a vigorous anti-apoptotic, pro-survival response to counteract mitochondrial apoptotic signaling. The heart up-regulates contractility of remaining myocytes in order to maintain cardiac output. We hypothesize that mutant mitochondrial proteins originate apoptotic signaling by interacting with proteins already in place in the mitochondrial outer membrane that regulate apoptosis, for example the pro-apoptotic protein Bak. Since it is unlikely that all mutant mitochondrial proteins have the necessary structure and localization within the inner membrane to activate Bak appropriately, only a small fraction of an age-associated burden of mtDNA mutations may be pathogenic. In this model, reactive oxygen species generated by mitochondrial respiration drive the formation of mtDNA mutations, but are not the primary mechanism for their pathogenicity. PMID- 16313222 TI - Cellular responses to protein accumulation involve autophagy and lysosomal enzyme activation. AB - Protein oligomerization and aggregation are key events in age-related neurodegenerative disorders, causing neuronal disturbances including microtubule destabilization, transport failure and loss of synaptic integrity that precede cell death. The abnormal buildup of proteins can overload digestive systems and this, in turn, activates lysosomes in different disease states and stimulates the inducible class of lysosomal protein degradation, macroautophagy. These responses were studied in a hippocampal slice model well known for amyloidogenic species, tau aggregates, and ubiquitinated proteins in response to chloroquine-mediated disruption of degradative processes. Chloroquine was found to cause a pronounced appearance of prelysosomal autophagic vacuoles in pyramidal neurons. The vacuoles and dense bodies were concentrated in the basal pole of neurons and in dystrophic neurites. In hippocampal slice cultures treated with Abeta(142), ultrastructural changes were also induced. Autophagic responses may be an attempt to compensate for protein accumulation, however, they were not sufficient to prevent axonopathy indicated by swellings, transport deficits, and reduced expression of synaptic components. Additional chloroquine effects included activation of cathepsin D and other lysosomal hydrolases. Abeta(142) produced similar lysosomal activation, and the effects of Abeta(142) and chloroquine were not additive, suggesting a common mechanism. Activated levels of cathepsin D were enhanced with the lysosomal modulator Z-Phe-Ala-diazomethylketone (PADK). PADK-mediated lysosomal enhancement corresponded with the restoration of synaptic markers, in association with stabilization of microtubules and transport capability. To show that PADK can modulate the lysosomal system in vivo, IP injections were administered over a 5 day period, resulting in a dose-dependent increase in lysosomal hydrolases. The findings indicate that degradative responses can be modulated to promote synaptic maintenance. PMID- 16313223 TI - A yang-invigorating Chinese herbal formula enhances mitochondrial functional ability and antioxidant capacity in various tissues of male and female rats. AB - The practice of traditional Chinese medicine (TCM) always emphasizes the prevention of diseases and delaying the onset of senility. In this regard, the maintenance of a balance of Yin and Yang-two opposing components involved in life activities as exemplified by the antagonistic action of the sympathetic and parasympathetic nervous systems-is essential in achieving a healthy condition. Previous studies have shown that long-term treatment with a Yang-invigorating Chinese herbal formula (VI-28) could increase red cell CuZn-superoxide dismutase (SOD) activity in male human subjects. In the present study, we examined the effects of chronic VI-28 treatment (80 and 240 mg/kg/day for 30 days) on red cell CuZn-SOD activity as well as mitochondrial functional ability and antioxidant components in various tissues of male and female rats. The results indicated that VI-28 treatment increased red cell CuZn-SOD activity as well as mitochondrial ATP generation capacity, reduced glutathione and alpha-tocopherol levels, and Mn-SOD activity in brain, heart, liver, and skeletal muscle tissues in both male and female rats to varying extents. The VI-28?induced increase in mitochondrial antioxidant capacity in various tissues was evidenced by the significant reduction in the extent of reactive oxygen species generation assessed by in vitro measurement. The red cell CuZn-SOD activities correlated positively with tissue mitochondrial antioxidant component levels/activity. The beneficial effect of VI-28 treatment on mitochondrial functional ability and antioxidant capacity may have clinical implications in the prevention of age-related diseases. PMID- 16313224 TI - NANOG changes in mouse kidneys with age. AB - NANOG is essential for mouse and human embryonic stem cell (ESC) pluripotency and selfrenewal. It is also expressed in several adult murine tissues as shown by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. However, human NANOG transcripts have been isolated from adult bone marrow (EST; GenBank accession no. BF893620). Here, we study the NANOG gene expression profile in isolated mouse renal papillary cells by Northern blot and RT-PCR. The whole RNA of mouse renal cells was obtained from fresh renal tissues, renal tissues infused by phosphate-buffered saline (PBS), and isolated renal papillary cells of mouse, respectively, as well as the renal papillary tissue from 18.5 days postcoitum (d.p.c.; fetal), 1-2-week-old (young), 1-8-month-old (adult), and 24-month-old (aging) mice. Our analysis shows that a very low expression level was detected in mouse renal tissues, and the renal papillary cells express more than other tissues as determined with Northern blot and RT-PCR. These data suggest that the kidney has its own cells expressing NANOG, and loss of NANOG expression occurs in an age-dependent manner in the kidney, either due to developmental factors or aging, particularly in renal papillary tissue. PMID- 16313227 TI - International Association of Biomedical Gerontology, 11th Congress (Aarhus, Denmark, August 13-16, 2005). PMID- 16313228 TI - Alliance for aging research. PMID- 16313229 TI - Validated supercentenarian cases aged 114 and above. PMID- 16313233 TI - Polymeric nanoparticles for oral delivery of drugs and vaccines: a critical evaluation of in vivo studies. AB - Oral drug delivery is the preferred route of administration of drugs. Because of their versatility, nanoparticles often have been investigated for the delivery of a wide number of drugs by this route. This article first examines the physicochemical, pharmaceutical and technological aspects that make nanoparticles a potential oral delivery system for drugs and active biomolecules. Next, upon consideration of in vivo studies, the pharmacokinetic, pharmacological and therapeutic aspects of orally administered nanoparticles are described. Special emphasis is placed on improvement of oral bioavailability of drugs incorporated into nanoparticles. Two main mechanisms involved in enhancing drug absorption are discussed: the protection of drug by nanoparticles against harsh conditions in the gut and the prolongation of gastrointestinal transit of nanoparticles by using bioadhesive polymers. Furthermore, nanoparticle uptake by intestinal cells and oral vaccination by these colloidal carriers are also covered. In this context, the immune responses elicited as well as the protection against pathogens induced by antigen-loaded nanoparticles administered by the oral route are presented. Finally, the main limitations and perspectives of these colloidal carriers as oral drug delivery systems are discussed. PMID- 16313234 TI - Pharmacogenomics and its potential impact on drug and formulation development. AB - Recent advances in genomic research have provided the basis for new insights into the importance of genetic and genomic markers during the different stages of drug development. A new field of research, pharmacogenomics, which studies the relationship between drug effects and the genome, has emerged. Structural pharmacogenomics maps the complete DNA sequences of whole genomes (genotypes) including individual variations, and functional pharmacogenomics assesses the expression levels of thousands of genes in one single experiment. Together, these two areas of pharmacogenomics have generated massive databases, which have become a challenge for the research field of informatics and have fostered a new branch of research, bioinformatics. If skillfully used, the databases generated by pharmacogenomics together with data mining on the Web promise to improve the drug development process in a variety of areas: identification of drug targets, evaluation of toxicity, classification of diseases, evaluation of formulations, assessment of drug response and treatment, post-marketing applications, and development of personalized medicines. PMID- 16313235 TI - A novel subfamily of monomeric inorganic pyrophosphatases in photosynthetic eukaryotes. AB - Two sPPases (soluble inorganic pyrophosphatases, EC 3.6.1.1) have been isolated from the microalga Chlamydomonas reinhardtii. Both are monomeric proteins of organellar localization, the chloroplastic sPPase I [Cr (Ch. reinhardtii)-sPPase I, 30 kDa] is a major isoform and slightly larger protein than the mitochondrial sPPase II (Cr-sPPase II, 24 kDa). They are members of sPPase family I and are encoded by two different cDNAs, as demonstrated by peptide mass fingerprint analysis. Molecular phylogenetic analyses indicated that Cr-sPPase I is closely related to other eukaryotic sPPases, whereas Cr-sPPase II resembles its prokaryotic counterparts. Chloroplastic sPPase I may have replaced a cyanobacterial ancestor very early during plastid evolution. Cr-sPPase II orthologues are found in members of the green photosynthetic lineage, but not in animals or fungi. These two sPPases from photosynthetic eukaryotes are novel monomeric family I sPPases with different molecular phylogenies and cellular localizations. PMID- 16313236 TI - Incorporation of partial polyhedrin homology sequences (PPHS) enhances the production of cloned foreign genes in a baculovirus expression system. AB - Baculovirus expression vector systems (BEVSs) have been used extensively for high level expression of cloned foreign genes. In many instances, the levels of recombinant protein(s) produced in insect cells and larvae are insufficient for experimental purposes. Thus new techniques and methods are needed to increase significantly the protein expression levels in BEVS. In the present paper, we describe the incorporation of a 15 bp element derived from the 5'-end partial sequence of the polyhedrin gene, which contains the non-coding sequence ATAAAT and the coding sequence ATGCCGAAT, into the 5'-end of the CTB (cholera toxin B subunit)-INS (insulin) fusion gene. With the addition of the PPHS (partial polyhedrin homology sequences), two extra amino acids (Pro-Asn) were added to the N-terminus of the mCTB-INS (modified CTB-INS) fusion protein. This new fusion protein was expressed in both insect cells and larvae using BEVSs. We found that the addition of PPHS enhanced 4-fold the expression of CTB-INS in both insect cells and larvae. Further analysis revealed that the additional two amino acids in mCTB-INS did not significantly affect binding affinity for G(M1) ganglioside. Therefore the PPHS can be used as a constitutive element immediately downstream of the polyhedrin promoter to induce significant increases in the expression levels of cloned foreign genes. PMID- 16313237 TI - Development and evaluation of a modified colorimetric solid-phase microassay for measuring the activity of cellular and plasma (Factor XIII) transglutaminases. AB - Plasma TG (transglutaminase) [FXIII (Factor XIII)] stabilizes fibrin and plays an essential role in haemostasis. In the present paper, we report a simple colorimetric assay for measuring FXIII activity. The advantage of this approach, compared with all the other solid-phase assays described so far for measuring TG activity, is that the first substrate, namely the synthetic dipeptide, N benzyloxycarbonyl-L-Gln-L-Gly, is coupled covalently by its C-terminus with amine substituted polystyrene plates. Covalent coupling eliminates the problem of desorption of proteins such as casein and N,N'-dimethylcasein (first substrates) when 'blocking' buffers containing proteins, e.g. BSA, are employed, or when samples of plasma or cell homogenates are used. The principle of the assay itself is based on the incorporation of the well-known second substrate, 5 (biotinamido)pentylamine, into the gamma-carboxamide of glutamine in the immobilized dipeptide. The amount of biotinylated amine bound to the plate, as measured by the phosphatase activity of Extravidin phosphatase attached to the biotin moiety, is directly proportional to the TG activity. The method shows strong correlation (r = 0.96) with the radiometric assay for FXIII activity. For plasma samples, a linear response was obtained in the range 0-1.33 i.u. (international unit)/ml, versus the Behring standard. In a preliminary clinical investigation, the method was applied to normal and pathological plasma samples from patients with liver failure and disseminated intravascular coagulation. In the isolation of TG from guinea-pig liver, it was used to measure enzyme activity after each purification step. This method is rapid, sensitive and can easily be applied to routine clinical analyses and to specific research problems. PMID- 16313238 TI - Narrow band UVB (311 nm), psoralen UVB (311 nm) and PUVA therapy in the treatment of early-stage mycosis fungoides: a right-left comparative study. AB - BACKGROUND: Psoralen ultraviolet A (PUVA) is a widely used first-line therapy for treatment of early cutaneous T-cell lymphoma. Narrow band UVB (UVB-NB) (311 nm) has been recently introduced as another effective line of treatment. It is postulated that the efficacy of UVB-NB could be enhanced by addition of psoralen. AIM: The aim of the present work was to compare the clinical and histopathologic efficacy of PUVA and UVB-NB in the treatment of early-stage MF (stages IA, IB and IIA), and to evaluate whether psoralen adds to the efficacy of UVB-NB or not. PATIENTS AND METHODS: Twenty patients (stage IA, IB or IIA) were divided into two equal groups: group I received UVB-NB on the right body half vs. PUVA on the left side of the body for 48 sessions, and group II received PUVB-NB on the right side of the body vs. PUVA on the left side for 36 sessions. The sessions were administered three times weekly. RESULTS: In group I, almost equal results were obtained on both sides, i.e., UVB-NB and PUVA were equally effective in the treatment of early stages of MF, both clinically and histopathologically. In group II, PUVB-NB was found to be as effective as conventional PUVA in the treatment of early-stage mycosis fungoides, also on both clinical and histopathological grounds. CONCLUSION: UVB-NB phototherapy should be included among the initial therapeutic options of mycosis fungoides in view of its efficacy, convenience, and likelihood of fewer long-term adverse effects. Addition of psoralen does not seem to enhance its therapeutic efficacy. PMID- 16313239 TI - Determination of action spectrum for sparfloxacin-photosensitized single-strand breaks in plasmid pBR322 DNA. AB - BACKGROUND: Various drugs have been reported to induce photosensitivity as a side effect. Sparfloxacin (SPFX) is well known to trigger dermatological phototoxicity upon solar radiation exposure. PURPOSE: To prevent SPFX-induced phototoxicity, we determined the wavelength range responsible for SPFX phototoxicity. METHODS: The action spectrum for SPFX photosensitization was assessed by the formation of single-strand breaks in plasmid pBR322 DNA. RESULTS: The wavelengths of light leading to the formation of single-strand breaks were in the ultraviolet A (UVA) and visible ranges. In comparison with the absorption spectrum, we found that SPFX absorption primarily contributed to the action spectrum of single-strand break formation, but it even expanded to the visible range (between 320 and 480 nm) beyond the absorption wavelengths. CONCLUSION: The findings suggest that protection of skin from short wavelengths of visible light beyond the absorption wavelengths as well as UVA light is of primary importance in prevention against induction of SPFX phototoxicity. PMID- 16313240 TI - Lymphocytes treated by extracorporeal photopheresis can down-regulate cytokine production in untreated monocytes. AB - BACKGROUND: Pro-inflammatory cytokines are actively involved in graft-versus host disease (GvHD) aetiology. Treatment of GvHD, using extracorporeal photopheresis (ECP), has demonstrated clinical efficacy. ECP rapidly reduces the number of T cells that produce tumour necrosis factor alpha (TNFalpha), interferon gamma (IFNgamma) and interleukin (IL)2. ECP-treated cells are re-infused immediately after completion of treatment. This study attempted to determine the influence that ECP-treated cells would have on untreated cells following re-infusion. METHODS: Heparinized samples were taken from 10 chronic GvHD patients, pre-ECP and immediately prior to re-infusion (post-ECP). Lymphocytes and monocytes were isolated using magnetic separation. The post-ECP lymphocytes were mixed with pre ECP monocytes, while the post-ECP monocytes and pre-ECP lymphocytes were combined. After suitable stimulation, the T cells were tested for intracellular TNFalpha, IFNgamma and IL2, while the monocytes were evaluated for TNFalpha, IL1alpha, IL1beta, IL6 and IL8. RESULTS: Although cytokine secretion is decreased in T cells exposed to ECP, pre-ECP T cells were unaffected by post-ECP monocytes. Post-ECP monocytes demonstrated a reduction in cytokine secretion. Furthermore, untreated monocytes down-regulated cytokine production following exposure to ECP treated lymphocytes. CONCLUSION: ECP has both a direct and indirect immunosuppressive action, both of which may be beneficial in the treatment of GvHD. PMID- 16313241 TI - Pre-vitamin D effective ultraviolet transmission through clothing during simulated wear. AB - BACKGROUND/PURPOSE: Clothing is an important protective layer used to reduce ultraviolet (UV) exposures to the skin surface. However, not all UV exposure is linked to detrimental health effects with some exposure to UVB wavelengths below 316 nm required for the synthesis of pre-vitamin D(3). The aim of the current research was to investigate the effect of fabric type, color, fit, and wetness on the transmission of pre-vitamin D(3) effective UV through garments during simulated wear, in a high UV exposure environment. METHODS: Dosimeters fabricated from polysulfone film were positioned at eight selected body sites on the skin surface and clothing surface of identically designed, loose and fitted, black and white T-shirts made up in two knitted fabric types and tested when both dry and when drying after initial wetting (n=3 replicates). The T-shirts were placed on manikins set to simulate humans in the sun between 09:30 and 12:30 Eastern Standard Time during the Southern Hemisphere summer period. The post-exposure absorbance was measured and the dosimeters were calibrated for biologically effective UV for pre-vitamin D(3) synthesis with a UV spectroradiometer. The effect of fit, fabric type, color, and wetness on pre-vitamin D(3) effective UV transmission during simulated wear was assessed. RESULTS: Irradiances varied among body sites with the highest erythemal exposures to a horizontal plane over the 3 h period reaching approximately 14.5 minimal erythema dose (MED) while the highest exposure under the garment was 0.22 MED which may not be above the threshold for pre-vitamin D(3) synthesis for the time period investigated. Fabric and fit were the main variables affecting transmission of pre-vitamin D(3) effective UV. Some interactions were identified between the fabric color and wetness and between fabric type and color; however, while significantly modifying transmission these effects were small. CONCLUSION: Transmission of pre-vitamin D(3) effective UV occurred through the high UPF knitted fabrics investigated. However, the length of exposure will influence whether the irradiances are sufficient to be above the threshold for pre-vitamin D(3) synthesis. The main effect on transmission of pre-vitamin D(3) effective UV was the fit of the T shirt and its fabric type (probably structure) rather than color or degree of wetness. PMID- 16313242 TI - Skin damage and mitochondrial dysfunction after acute ultraviolet B irradiation: relationship with nitric oxide production. AB - BACKGROUND: Ultraviolet (UV) radiation is the main environmental carcinogen. It is able to induce injury in the keratinocytes, which triggers mechanisms in order to protect the skin against molecular alterations that may lead to the development of skin cancer. UVB is capable of producing genotoxic damage, directly or indirectly through reactive oxygen species, inducing DNA alterations and mutations. UVB radiation has also been associated with the generation of nitric oxide (NO), which is able to induce many physiological and physiopathological processes. The aim of the current study was to investigate the effect of UVB irradiation in hairless mice skin. METHODS: We evaluated the effect of an acute dose (200 mJ/cm(2)) of UVB irradiation correlating with histological alterations, nitric oxide synthase expression and activity, mitochondrial respiratory function, superoxide anion production and lipid peroxidation, 0, 6, 17 and 24 h post-irradiation treatment. RESULTS: Morphological analysis showed disruption of the epidermal stratum corneum and basale after UVB irradiation. The results indicated that skin UVB irradiation was associated with an increased cytosolic inducible nitric oxide synthase (iNOS) expression, inversely related to lipid peroxidation processes. An increase in mitochondrial superoxide anion (O(2) (*-)) and NO production 17 h post-irradiation was correlated with a mitochondrial dysfunction, all of them integrating the skin response to acute UVB irradiation. CONCLUSIONS: UVB irradiation of the skin produces morphological alterations as a consequence of the induction of molecular mechanisms associated with mitochondrial respiratory dysfunction and O(2) (*-) production, probably mediated by the increased mitochondrial NO production. On the other hand lipid peroxidation decrease inversely correlates with cytosolic iNOS expression, suggesting a protective role for the inflammatory response. PMID- 16313243 TI - Experimental phytophotodermatitis. AB - BACKGROUND/AIMS: Phytophotodermatitis (PPD) is defined as a phototoxic reaction of the skin after contact with substances derived from plants and subsequent exposure to sunlight. It is a frequent disease in our outpatient clinics during summer because of contact with Tahitian lemon. Our objectives were to experimentally reproduce PPD in rats, to identify whether PPD is induced by minimal exposure periods to sunlight, to find what kinds of lemons and which parts of the lemon (the fruit juice or the peel juice) may trigger the disease; to know whether the use of sunblock prevents the reaction; and to perform light microscopy of the lesions to describe their histology. METHODS: Adult rats (Rattus norwegicus), three in each experiment, were used. After painting the rats with the fruit juice or the peel juice they were exposed to sunlight for 2.5, 5, 7.5, and 10 min. Tahitian and Sicilian lemons were used in the experiments. Biopsies with 3-mm punches of different times of exposure were performed. RESULTS: The peel juice of both lemons reproduced PPD, which was clinically evident after 48 h. When the peel juice was alone applied there was no reaction; moreover, exposure to sunlight alone triggered no reaction. Two and a half minutes of exposure time was sufficient to induce phototoxic reaction, which was time dependent (the longer the exposure the more intense the reaction). Histopathological studies showed epithelial time-dependent vacuolar degeneration. The use of sunblock diminished the intensity of the reaction but did not prevent it. CONCLUSION: PPD can be reproduced in an animal model. It may be caused by the peel juice of Tahitian and Sicilian lemon. Because of an extremely short time of exposure (2.5 min) is sufficient to induce PPD it is necessary to alert the population, of the need for caution when handling lemons, especially outdoors despite using sunblock. PMID- 16313244 TI - Topical aminolevulinic acid-photodynamic therapy for the treatment of acne vulgaris. AB - BACKGROUND: Photodynamic therapy (PDT) is useful for the treatment of epidermal neoplasia but may also have use in the treatment of inflammatory dermatoses. The purpose of this study was to evaluate the safety and efficacy of PDT in the treatment of acne on the face. METHODS: Eight patients with mild-to-moderate acne vulgaris on the face were treated with one session of topical aminolevulinic acid (ALA)-PDT using a red light. Each patient's face was divided into two areas (right side and left side) for ALA-PDT and untreated control. Twenty percent topical ALA was applied with 4 h occlusion, and 18 J/cm(2) red light (630+/-63 nm) was administered. Each patient's acne was visually assessed by a spot count of both inflamed and non-inflamed lesions at baseline and at 1, 3, and 6 months following PDT. RESULTS: At 1, 3, and 6 months, the mean percentage reduction in inflamed lesions count was 27.6%, 37.9%, and 41.9%, respectively, in the ALA-PDT treated area, whereas in the control area, it was 8.0%, 14.7%, and 15.4%, respectively, compared with the baseline. This showed a higher reduction of a spot count of inflamed lesions in the ALA-PDT-treated area, which was statistically significant (P<0.05). There was a slightly higher reduction of a spot count of non-inflamed lesions in the ALA-PDT-treated area, which was statistically not significant (P>0.05). The adverse effects were minimal. CONCLUSION: ALA-PDT with red light was found to be effective in the reduction of the number of inflamed lesions in patients with mild-to-moderate acne vulgaris, with no significant short-term adverse effects. PMID- 16313245 TI - Clinical and immunohistochemical evaluation of psoriatic plaques treated with topical 5-aminolaevulinic acid photodynamic therapy. AB - BACKGROUND/PURPOSE: The aims of this study were to investigate the clinical and immunohistochemical events of psoriatic plaques during photodynamic therapy (PDT) using topical application of 5-aminolaevulinic acid (ALA). METHODS: Twelve psoriatic patients were recruited for this study. Four of them dropped out because of pain during treatment. The effect of PDT was evaluated in the remaining eight. One plaque was selected in each patient and treated once weekly with PDT 10-30 J/cm(2) two to five times. It was evaluated by using the scale, erythema and induration (SEI) index (maximal score per patient=9). Pain during treatment was assessed by a visual analogue scale (VAS), ranging from 0 to 10. Skin biopsies were taken before treatment, after two treatments and after completion of treatment, and were evaluated by immunohistochemistry. RESULTS: Median SEI scores were significantly reduced from 7 (range 5-9) before to 1.5 (range 0-3) following treatment (P<0.0001). The median pain during PDT was 7. The number of vessels in the subpapillary dermis, identified by antibodies against Factor VIII and endoglin, increased during and/or after treatment in six of eight patients. Before treatment, the epidermal growth factor (EGF) receptor was displayed throughout the epidermis, keratin 16 suprabasally, involucrin from the stratum granulosum to the lower spinous layers and filaggrin in stratum granulosum with focal absence. There was a moderate dermal infiltrate of CD4(+) cells and a sparse one of CD8(+). Following treatment, the EGF receptor was still displayed throughout the epidermis in seven of eight specimens. Cytokeratin 16 expression decreased markedly. Involucrin was not seen as deep in the spinous layers as before PDT. Filaggrin was expressed throughout the stratum granulosum and often weakly in the upper stratum spinosum. The number of CD4(+) and CD8(+) dermal cells decreased. CONCLUSION: PDT improved psoriasis and induced dermal neovascularization. Although a good clinical response was seen in most of our patients, the high frequency of discomfort during treatment limits the usefulness of ALA-PDT for psoriasis. The mechanism of the neovascularization is unknown. It may be owing to an indirect effect of PDT on the microvasculature and immune system or recovery phenomena. PMID- 16313246 TI - Does narrow-band ultraviolet B phototherapy work in atopic dermatitis through a local or a systemic effect? AB - The likely mechanisms of action of narrow-band ultraviolet B (NB-UVB) in atopic dermatitis are several. We attempted, in a 12 patient prospective intraindividually controlled study, to determine whether the effect of NB-UVB in atopic dermatitis is primarily through systemic or local effects. Change in observer-assessed severity of patches of dermatitis covered during each whole body NB-UVB treatment was compared with change in uncovered neighbouring patches. We found great variation between patients in responses. Only in two (of 12) patients was there a large difference between directly exposed and covered patch dermatitis severity. We suspect that the balance of local and systemic effects important in determining response to NB-UVB in atopic dermatitis varies from patient to patient. This study did not conclusively answer our original question, but did show that in some patients localized effects of NB-UVB are important. PMID- 16313247 TI - Frailty predicts long-term mortality in elderly subjects with chronic heart failure. AB - BACKGROUND: The elderly are characterized by a high prevalence of chronic heart failure (CHF) and frailty, which is a complex interaction of physical, psychological and social impairment. This study aimed to examine the predictive role of frailty on long-term mortality in elderly subjects with CHF. MATERIALS AND METHODS: The study assessed long-term mortality after 12-year follow up in 120 subjects with CHF and 1139 subjects without CHF, selected in 1992, from a random sample of the elderly population in the Campania region of Italy. Frailty was assessed according to a 'Frailty Staging System'. RESULTS: Subjects with CHF were prevalently female (60%) and older than 75 years (mean 75.9 +/- 6.7); subjects without CHF were prevalently female (56.4%) and younger than 75 years (mean 74.0 +/- 6.3). In subjects with and without CHF stratified into classes of frailty there was a statistically significant increase in age, comorbidity, disability and low social support, and a decrease in MMSE score. Moreover, death progressively increased more with frailty in subjects (70.0% to 94.4%, P < 0.03) than in those without (43.8.% to 88.3%, P < 0.0001) CHF. The Kaplan-Meier analysis shows that at 9 years the probability of survival progressively decreased as frailty increased (45.5% to 0%) in subjects with CHF and from 62.8% to 25.9% in subjects without CHF. The Cox regression analysis indicated that frailty is predictive of mortality in the multivariate model adjusted for several variables including sex and age in subjects with and without CHF. Moreover, the analysis showed that frailty is more predictive of mortality in elderly subjects with CHF when it was analyzed either as continuous (1.48 vs. 1.36) or as a dummy (3 vs. 1 = 1.62 vs. 1.24) variable. CONCLUSIONS: Thus mortality among elderly subjects with or without CHF increases with frailty. Moreover, frailty is more predictive of long-term mortality in elderly subjects with than in those without CHF. Hence, frailty represents a new independent variable for predicting long term mortality in elderly subjects with CHF. PMID- 16313248 TI - Haem oxygenase-1 genotype and cardiovascular adverse events in patients with peripheral artery disease. AB - BACKGROUND: A functional GT dinucleotide length polymorphism in the haem oxygenase-1 (HO-1) gene promoter is thought to be involved in the pathogenesis of cardiovascular disease. Short (< 25) (GT)n repeats are suggested to facilitate enhanced HO-1 up-regulation in response to injury and confer potent anti inflammatory and antioxidative effects. MATERIALS AND METHODS: We investigated the association between the HO-1 GT-polymorphism and cardiovascular outcome in 472 patients with advanced peripheral artery disease. Cardiovascular risk profile and DNA samples for determination of the HO-1 genotype (carrier vs. noncarrier of a short (GT)n repeat allele) were obtained at baseline, and patients were followed for median 21 months for the occurrence of coronary events (myocardial infarction, percutaneous coronary interventions and coronary artery bypass graft), cerebrovascular events (stroke or carotid revascularization) and all cause mortality. RESULTS: Coronary events occurred in 48 patients (9%), cerebrovascular events in 40 patients (9%) and 59 patients (13%) died. In total, 173 major adverse cardiovascular events (MACE) occurred in 133 patients (28%). Carriers of the short (GT)n repeat allele had a 0.46-fold reduced adjusted hazard ratio for coronary events (P = 0.016) as compared to noncarriers. No significant difference was found for cerebrovascular events, mortality and overall MACE. CONCLUSION: Apparently, the HO-1 genotype exerts potentially protective effects against coronary adverse events in patients with peripheral artery disease. Homozygous and heterozygous carriers of < 25 (GT)n repeats had lower rates of myocardial infarction, percutaneous coronary interventions and coronary bypass operations compared to patients with longer (GT)n repeats. PMID- 16313249 TI - The effect of exercise on large artery haemodynamics in healthy young men. AB - BACKGROUND: Brachial blood pressure predicts cardiovascular outcome at rest and during exercise. However, because of pulse pressure amplification, there is a marked difference between brachial pressure and central (aortic) pressure. Although central pressure is likely to have greater clinical importance, very little data exist regarding the central haemodynamic response to exercise. The aim of the present study was to determine the central and peripheral haemodynamic response to incremental aerobic exercise. MATERIALS AND METHODS: Twelve healthy men aged 31 +/- 1 years (mean +/- SEM) exercised at 50%, 60%, 70% and 80% of their maximal heart rate (HRmax) on a bicycle ergometer. Central blood pressure and estimated aortic pulse wave velocity, assessed by timing of the reflected wave (T(R)), were obtained noninvasively using pulse wave analysis. Pulse pressure amplification was defined as the ratio of peripheral to central pulse pressure and, to assess the influence of wave reflection on amplification, the ratio of peripheral pulse pressure to nonaugmented central pulse pressure (PPP : CDBP-P1) was also calculated. RESULTS: During exercise, there was a significant, intensity-related, increase in mean arterial pressure and heart rate (P < 0.001). There was also a significant increase in pulse pressure amplification and in PPP : CDBP-P(1) (P < 0.001), but both were independent of exercise intensity. Estimated aortic pulse wave velocity increased during exercise (P < 0.001), indicating increased aortic stiffness. There was also a positive association between aortic pulse wave velocity and mean arterial pressure (r = 0.54; P < 0.001). CONCLUSIONS: Exercise significantly increases pulse pressure amplification and estimated aortic stiffness. PMID- 16313250 TI - Strength and endurance training lead to different post exercise glucose profiles in diabetic participants using a continuous subcutaneous glucose monitoring system. AB - BACKGROUND: Although both strength training (ST) and endurance training (ET) seem to be beneficial in type 2 diabetes mellitus (T2D), little is known about post exercise glucose profiles. The objective of the study was to report changes in blood glucose (BG) values after a 4-month ET and ST programme now that a device for continuous glucose monitoring has become available. MATERIALS AND METHODS: Fifteen participants, comprising four men age 56.5 +/- 0.9 years and 11 women age 57.4 +/- 0.9 years with T2D, were monitored with the MiniMed (Northridge, CA, USA) continuous glucose monitoring system (CGMS) for 48 h before and after 4 months of ET or ST. The ST consisted of three sets at the beginning, increasing to six sets per week at the end of the training period, including all major muscle groups and ET performed with an intensity of maximal oxygen uptake of 60% and a volume beginning at 15 min and advancing to a maximum of 30 min three times a week. RESULTS: A total of 17,549 single BG measurements pretraining (619.7 +/- 39.8) and post-training (550.3 +/- 30.1) were recorded, correlating to an average of 585 +/- 25.3 potential measurements per participant at the beginning and at the end of the study. The change in BG-value between the beginning (132 mg dL( 1)) and the end (118 mg dL(-1)) for all participants was significant (P = 0.028). The improvement in BG-value for the ST programme was significant (P = 0.02) but for the ET no significant change was measured (P = 0.48). Glycaemic control improved in the ST group and the mean BG was reduced by 15.6% (Cl 3-25%). CONCLUSION: In conclusion, the CGMS may be a useful tool in monitoring improvements in glycaemic control after different exercise programmes. Additionally, the CGMS may help to identify asymptomatic hypoglycaemia or hyperglycaemia after training programmes. PMID- 16313251 TI - Gestational diabetes mellitus is associated with increased C-reactive protein concentrations in the third but not second trimester. AB - OBJECTIVE: Serum C-reactive protein (CRP) concentrations were measured longitudinally throughout pregnancy to test the hypothesis that CRP could relate more closely to glucose tolerance than to adiposity. METHODS: The CRP concentrations in pregnant women with normal glucose tolerance (NGT) and those with gestational diabetes mellitus (GDM) were measured at the same time as the oral glucose tolerance test (OGTT), at the 24th and 28th weeks of gestation and between the 37th and 38th weeks of gestation. RESULTS: At the end of the third trimester, women with GDM had significantly higher CRP levels than women with NGT [median (interquartile range), 9.7 mg L(-1) (5.4-16.0) and 5.7 mg L(-1) (5.1 7.2); P < 0.001, respectively], but at the time of the diagnostic OGTT no significant difference between the two groups was observed. This was owing to a significant increase of CRP in women with GDM between the time of the OGTT and the 37th-38th gestational weeks [median (interquartile range), 1.9 mg L(-1) ( 2.2, 6.7); P = 0.01]; whereas, no change in CRP was found in women with NGT [median (interquartile range), -0.1 mg L(-1) (-2.4, 3.1); P = 0.76]. Multiple linear regression analysis showed only a significant independent influence of GDM (P < 0.001) on maternal CRP concentrations in the 37th-38th gestational weeks and a significant influence of body mass index (P < 0.007), but no influence of GDM at the time of the OGTT. CONCLUSION: These data suggest that in women with gestational diabetes the CRP concentration is primarily related to the degree of adiposity until the second trimester and that thereafter impaired glucose metabolism appears to be the predominant predictor of changes in CRP. PMID- 16313252 TI - Chorionic plate artery function and Doppler indices in normal pregnancy and intrauterine growth restriction. AB - BACKGROUND: In fetal growth restriction (FGR) abnormal umbilical artery (UA) Doppler waveform indices suggest increased vascular resistance and impaired placental blood flow. This study aimed to determine whether UA Doppler waveform indices were related to the vasoreactivity of placental chorionic plate small arteries in normal and FGR pregnancies. MATERIALS AND METHODS: UA Doppler waveform analysis was performed 24 h before delivery in 23 normal term and 15 FGR pregnancies. Post-delivery responses of chorionic plate arteries to vasoactive agents were examined using the technique of wire myography. RESULTS: Altered vascular reactivity to agonists was demonstrated in chorionic plate arteries in FGR pregnancy. Constriction to U46619 (thromboxane mimetic) and relaxation to sodium nitroprusside (nitric oxide donor) were significantly increased in the arteries of FGR pregnancies compared with normal pregnancies. No relationship existed between Doppler indices and chorionic plate responses in normal or FGR pregnancies. CONCLUSIONS: Fetoplacental vascular reactivity is altered in FGR pregnancy independently of UA Doppler waveform indices. Altered function may be additive to the pathophysiology underlying abnormal Doppler waveforms and could contribute to the inappropriate control of vascular tone in FGR. PMID- 16313253 TI - Modulatory effect of mannose-binding lectin on cytokine responses: possible roles in HIV infection. AB - BACKGROUND: Mannose-binding lectin (MBL) is a soluble receptor of the innate immune system, probably contributing to antimicrobial defence. The possible role of MBL in HIV infection is unclear. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) from 28 HIV-infected patients and 13 healthy controls were stimulated with MBL and costimulated with HIV-1 gp120 or mannan from Saccharomyces cerevisiae before inflammatory responses in PBMC cultures were examined by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. HIV-1 RNA replication in vitro was assessed by quantitative reverse transcription polymerase chain reaction (RT-PCR) in supernatants from patients with measurable HIV-1 RNA levels. RESULTS: (i) Enhanced TNF-alpha responses were observed when PBMCs from healthy controls and HIV-infected patients were stimulated with MBL and costimulated with HIV-1 gp120 or mannan. (ii) MBL stimulation induced increased HIV RNA replication in culture supernatants when costimulated with mannan. CONCLUSIONS: The present study suggests a modulatory role of MBL on cytokine responses, and HIV replication after stimulation with microbial products. These effects of MBL on inflammatory responses and viral replication may be clinically relevant for HIV infection. PMID- 16313255 TI - Acquired C1-inhibitor deficiency in a patient with systemic lupus erythematosus: a case report and review of the literature. AB - The C1-inhibitor (C1-INH) is an important member of the serpin family which inhibits the first component of the human complement system and controls contact activation of the coagulation and kinin system. An acquired form of C1-INH deficiency was recognized and classified as type I, which is characterized by accelerated catabolism of C1-INH, whereas type II is defined by the presence of an autoantibody directed against the C1 inhibitor molecule. This study reports the case of a 32-year-old woman with systemic lupus erythematosus (SLE) who experienced recurrent angioedema because of an acquired C1-INH deficiency. The relevant literature is reviewed. PMID- 16313254 TI - Tumour necrosis factor alpha in fat redistribution syndromes associated with combination antiretroviral therapy in HIV-1-infected patients: potential role in subcutaneous adipocyte apoptosis. AB - BACKGROUND: The pathogenesis of fat redistribution syndromes (FRS) observed in the setting of highly active antiretroviral therapy (HAART) for the treatment of HIV-1-infection remains elusive. A dysregulation of the tumour necrosis factor alpha (TNF-alpha) system occurs in HIV-infected patients with FRS. MATERIALS AND METHODS: The study looked at both the in vivo and in vitro relationship between TNF-alpha and the degree of subcutaneous adipocyte apoptosis in 60 HIV-1-infected patients on HAART with FRS, another 60 HIV-1-infected patients on HAART without FRS and 60 uninfected control patients. Apoptosis was assessed by the terminal deoxynucleotidyl transferase dUTP (deoxyuridine 5'-triphosphate)-digoxigenin Nick End Labelling (TUNEL) method. Soluble receptors of TNF-alpha were determined by the sandwich enzyme immunoassay technique. The in vitro viability was assessed by staining with 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) and apoptosis by TUNEL. RESULTS: HIV-1-infected patients with FRS had significantly higher degrees of subcutaneous adipocyte apoptosis than those without FRS (P = 0.0001) and uninfected controls (P < 0.0001). There was a statistically significant association between serum levels of soluble TNF-alpha receptors #1 and #2 and the degree of subcutaneous adipocyte apoptosis in patients with and without FRS (P < 0.0001 for both receptors). In vitro, the addition of TNF-alpha (10 ng mL(-1)) to an adipocyte culture embedded with indinavir, either alone or in clinically relevant combinations with stavudine (d4T) and lamivudine (3TC), significantly decreased adipocyte viability (P = 0.0001) and increased adipocyte apoptosis (P < 0.0001) with respect to that observed with the addition of antiretrovirals alone. CONCLUSIONS: TNF-alpha plays a significant role in subcutaneous adipocyte apoptosis, which occurs in the setting of FRS in HIV-1-infected patients on highly active antiretroviral therapy. PMID- 16313256 TI - Alterations in electrolyte equilibrium in patients with acute leukemia. AB - BACKGROUND AND AIM: A wide array of disturbances in electrolyte equilibrium is commonly seen in patients with acute leukemia (AL). These abnormalities present a potential hazard in these patients, as that of enhancing the cardiotoxic effects of certain chemotherapeutic regimens. The literature dealing with AL-related electrolyte abnormalities and their interactions in leukemic patients was reviewed. DATA SYNTHESIS: Sources included MEDLINE and EMBASE. The search strategy was based on the combination of 'acute leukemia', 'electrolyte abnormalities', 'acid-base disorders', 'potassium', 'sodium', 'magnesium', 'calcium', and 'phosphorus'. References of retrieved articles were also screened. A decrease in serum potassium, mainly owing to lysozyme-induced tubular damage, appears to be one of the most frequent and potentially hazardous abnormalities. Other clinically significant metabolic perturbations include hyponatremia and hypercalcemia. CONCLUSION: A broad spectrum of electrolyte abnormalities is encountered in the clinical setting of AL, which are related to the disease process per se and/or to the therapeutic interventions. Clinicians should be vigilant for early detection and appropriate management of these disorders before the initiation of chemotherapy regimens as well as during treatment. PMID- 16313257 TI - Human hematopoietic stem/progenitor-enriched CD34(+) cells are mobilized into peripheral blood during stress related to ischemic stroke or acute myocardial infarction. AB - The hematopoietic and non-hematopoietic stem/progenitor cells harvested directly from the bone marrow (BM) or G-CSF mobilized peripheral blood were demonstrated to play an important role in regeneration of damaged organs (1, 2). Here, we asked if the stroke- or acute heart infarct-related stress triggers mobilization of stem/progenitor-enriched CD34(+)cells from the BM into the peripheral blood, which subsequently could contribute to regeneration of damaged tissues. To address this question the peripheral blood samples were harvested from patients with ischemic stroke during the first 24 h of manifestation of symptoms and on the second and sixth day afterwards or during the first 24 h of acute cardiac pain as well as on the second and sixth day of infarct. We measured in these patients (i) percentage of circulating hematopoietic stem/progenitor-enriched CD34(+) cells in peripheral blood by employing fluorescence activated cell sorter (FACS) and (ii) number of hematopoietic progenitor cells for the granulocyte monocytic colony-forming unit (CFU-GM) and erythoid burst-forming unit (BFU-E) lineages circulating in peripheral blood. We concluded that stress related to ischemic stroke or acute myocardial infarction triggers the mobilization of hematopoietic stem/progenitor-enriched CD34(+) cells from the BM into peripheral blood. These circulating stem/progenitor-enriched CD34(+) cells may contribute to the regeneration of ischemic tissues, however, this possibility requires further studies. PMID- 16313258 TI - Increased population of CD4(+)CD25(high), regulatory T cells with their higher apoptotic and proliferating status in peripheral blood of acute myeloid leukemia patients. AB - PURPOSE: Regulatory T cells (T-reg) that control harmful autoimmune T cells in the periphery may also suppress the immune response against cancer. In this study we investigated the possible involvement of CD4(+)CD25(high) T-reg in the immune impairment of patients with acute myeloid leukemia (AML). EXPERIMENTAL DESIGN: The frequencies and phenotypes of CD4(+)CD25(high) T cells in the peripheral blood of AML patients were determined by flow cytometry. To assess the functional activity of CD4(+)CD25(high) T cells, CD4(+)CD25(high), and CD4(+)CD25(-) T cells were sorted from peripheral blood mononuclear cells with FACS Vantage. The immunoregulatory properties of CD4(+)CD25(high) and CD4(+)CD25(-) T cells were characterized by proliferation assays and cytokine production assays. In addition, the frequency of apoptotic and proliferating cells in CD4(+)CD25(high) T cells were respectively evaluated by 7AAD and ki67 binding cells using flow cytometry. RESULTS: Compared with healthy controls, AML patients had a higher proportion of CD4(+)CD25(high) T cells in peripheral blood. These cells were CD45 RA(-), CD69(-), CD45-RO(+), CD95(+), and intercellular CTLA-4(+), and secreted low levels of TNF-alpha and IL-10, but no IL-2, IL-4, IL-5, and IFN-gamma. They inhibited the proliferation and cytokine production (IL-2, IFN-gamma) of CD4(+)CD25(-) T cells, but improved IL-10 production under the co-culture of both subsets with stimulation, thus behaving as T-reg. Notably, CD4(+)CD25(high) T cells in AML patients presented significantly higher apoptosis and proliferation than that of healthy individuals. CONCLUSIONS: The frequency of CD4(+)CD25(high) T-reg in peripheral blood in AML patients is significantly higher when compared with healthy individuals, likely due to the increasing proliferation of CD4(+)CD25(high) T cells. PMID- 16313259 TI - Expression of poliovirus receptor-related proteins PRR1 and PRR2 in acute myeloid leukemia: first report of surface marker analysis, contribution to diagnosis, prognosis and implications for future therapeutical strategies. AB - Poliovirus receptor-related (PRR) proteins belong to the Nectin-adhesion molecules' group, are expressed on endothelial cells and on CD34(+) stem cells and mediate the organization of endothelial and epithelial junctions. There is evidence to suggest, that those receptors could have a role in leukemia. We have studied the expression of PRR molecules PRR1 and PRR2 on mononuclear bone marrow (BM) cells of 55 patients with acute myeloid leukemia (AML) at first diagnosis by FACS-analysis using directly Phycoerythrin-labeled markers (PRR1 clone R1.302.12; PRR2 clone R2.477.1) in combination with other Fluorescein conjugated antibodies to evaluate the blast phenotype in AML. The leukemic gate included blasts and residual monocytes and lymphocytes. A case was defined as positive, if more than 20% of the gated cells expressed the regarding receptor. We could demonstrate, that on average 35% PRR1(+) or 45% PRR2(+) cells in AML were found. Within FAB types we observed a high PRR1 expression in cases with M3 and M4 and lowest expressions in M0 and M5; a high PRR2 expression was found in cases with M3, M4, M5 and M1 and lowest expressions in M0 and M2. Separating our patients' cohorts in cytogenetic risk groups we could detect a significant higher proportion of PRR1(+) cases (73% vs. 25% of cases, P = 0.009) or PRR1(+) cells (57% vs. 18% of cases, P = 0.001) in the cytogenetic favorable risk vs. poor risk group (75% vs. 32% PRR2(+) cases). Moreover cut-off-values with a maximum probability for a significant differentiation between cases with higher or lower levels of these markers could be found: cases with >78% PRR1(+) and cases with >77% PRR2(+) cells were characterized by a tendency for longer relapse free survival times. Qui square analyses showed, that 3 of 4 cases with FAB-type M3 (P = 0.03) or a favorable karyotype (P = 0.04) were found in the group with >7% PRR1(+) cells, due to only few cases available a similar correlation, however, could not be found in cases with >78% PRR2(+) cells. We can conclude, that blasts in AML regularly express PRR1 and PRR2. Cases with a high expression of PRR1 or PRR2 are characterized by a more favorable prognosis. With respect to the individual PRR status the benefit of biological response modifiers as priming agents, differentiation mediators or factors influencing cellular metabolisms inducing factors can be discussed under a new point of view. PMID- 16313260 TI - Over-expression of tumor necrosis factor-alpha in bone marrow biopsies from patients with myelodysplastic syndromes: relationship to anemia and prognosis. AB - OBJECTIVES: An excessive intramedullar progenitor cell apoptosis, to which elevated expression of tumor necrosis factor-alpha (TNF-alpha) might contribute, is considered the main cause of inefficient hematopoiesis in myelodysplastic syndromes (MDS). Enhanced bone marrow (BM) angiogenesis is regarded as an essential cofactor in the progression of MDS to acute myelogenous leukemia (AML) and microvessel formation may be induced by TNF-alpha as well. To investigate TNF alpha signaling and neoangiogenesis as potential molecular pathways for therapeutic intervention in MDS with respect to the various MDS subtypes, we performed a morphological and clinico-pathological study on a large series of paraffin-embedded trephine BM biopsies. METHODS: TNF-alpha expression and BM vessels were immunohistochemically analyzed on 89 paraffin-embedded BM biopsies from patients with MDS and secondary AML, including 12 control samples. Data were correlated with clinico-pathological and laboratory parameters and analyzed for their prognostic significance considering overall survival. RESULTS: TNF-alpha was over-expressed in MDS patients, especially in those with refractory anemia and its expression correlated with BM cellularity and with magnitude of anemia as well as with microvessel density (MVD). TNF-alpha over-expression was associated with premature deaths. MVD was increased in MDS and secondary AML and correlated with marrow cellularity and expression of TNF-alpha, but was not of prognostic significance. CONCLUSIONS: TNF-alpha expression and MVD are elevated in MDS and secondary AML. TNF-alpha expression in BM progenitor cells appears to negatively impact erythropoiesis and overall survival in MDS, and may serve as a potential therapeutic target in patients with hypercellular MDS with marked anemia. PMID- 16313261 TI - Circulating endothelial cells in patients with acute myeloid leukemia. AB - OBJECTIVES: The circulating endothelial cells (CEC) are proposed to be a non invasive marker of angiogenesis. The level of CEC in peripheral blood (PB) of acute myeloid leukemia (AML) patients has not been investigated prior to this study. We evaluated the count of resting (rCEC), activated (aCEC) and endothelial progenitor cells (CEPC) in the PB of AML and healthy subjects. In addition we correlated the levels of CEC with disease status, known prognostic factors and response to treatment. METHODS: CEC were quantified by utilizing four-color flow cytometry procedures in 48 AML patients at the time of diagnosis and 29 healthy controls. Additionally, measurements were again taken after the first course of induction treatment in 12 of the patients. RESULTS: The numbers of aCEC, rCEC and CEPC were significantly higher in the AML patients than in the controls (P < 0.0001, P < 0.0001 and P < 0.001, respectively). The CEC count was significantly higher in the AML patients with white blood cell count (WBC) >15 G/L, elevated lactic dehydrogenase (LDH) levels and a higher (over median) absolute blasts count (ABC) in PB than in the group with WBC <15 G/L (P < 0.03), a normal LDH level (P < 0.03) and a lower (10%) and very low (0-1%) repp86 expression (P < 0.0001) in the tumor cells. The multivariate analysis revealed repp86 expression to be superior to other clinical characteristics as a prognostic factor (P = 0.0016). CONCLUSION: Based on these findings, repp86 expression is a new important prognostic factor in MCL. PMID- 16313263 TI - Methylation status of fragile histidine triad (FHIT) gene and its clinical impact on prognosis of patients with multiple myeloma. AB - Aberrant methylation of tumor suppressor genes (TSG) has been studied in multiple myeloma (MM). We determined the methylation status of the FHIT (fragile histidine triad) gene, a putative TSG, in 48 patients with MM. Clinical association with its methylation status was then analyzed. The FHIT gene methylation was observed in 21 of the 48 patients (44%). No association between FHIT gene methylation and clinical variables such as age, gender and clinical stage was found. However, the estimated 50% survival time of the methylated group was significantly shorter than that of the unmethylated group (18.2 vs. 45.1 months, P < 0.05). Univariate analysis revealed adverse prognostic factors: FHIT gene methylation (P = 0.028), poor performance status (I to IV, P = 0.002), anemia (< or =8.5 g/dL, P = 0.007), hypoalbuminemia (< or =3.5 g/dL, P < 0.002), high serum C-reactive protein levels (>0.5 mg/dL, P = 0.002), elevated beta-2-microglobulin serum levels (>6.5 mg/L, P < 0.001), and treatments not including autologous peripheral blood stem cell transplantation (auto-PBSCT) (P = 0.007). Multivariate analysis identified FHIT gene methylation [hazard ratio (HR) 1.722, 95% confidence interval (CI) 1.150 2.603, P = 0.009], elevated beta-2-microglobulin serum levels (>6.5 mg/L, HR 2.005, 95% CI 1.035-3.937, P = 0.004), and treatments not including auto-PBSCT are independent predictive variables. These findings indicate that aberrant methylation of the FHIT gene is an independent adverse prognostic factor in MM. PMID- 16313264 TI - Cardiac presentation of ALK positive anaplastic large cell lymphoma. AB - Cardiac involvement as an initial presentation of malignant lymphoma is a rare occurrence. We report the case of an immunocompetent 29-year-old male who presented with syncope and arrythmias secondary to a ventricular cardiac mass. Transcutaneous cardiac biopsy was non-diagnostic, therefore an open cardiac biopsy was performed from which a provisional diagnosis of a cardiac inflammatory pseudotumour was made. Six months after presentation, he developed several subcutaneous lesions with systemic symptoms. Histological and immunophenotypic review of the initial cardiac biopsy revealed features consistent with a diagnosis of CD30, ALK1 positive anaplastic large cell lymphoma (ALCL). Despite intensive treatment with combination chemotherapy, there was significant progression of disease, and he died 11 months after diagnosis. The overall prognosis of cardiac lymphoma remains poor, which may be due to the often late presentation of the tumour. To our knowledge, this is the first reported case of a cardiac ALK positive ALCL. Although rare, cardiac presentation of ALCL should be added to the list of differential diagnoses of cardiac lymphomas. PMID- 16313265 TI - A case of advanced Hodgkin lymphoma with breast involvement treated with a combination of gemcitabine and pegylated liposomal doxorubicin. AB - Gemcitabine in combination with pegylated liposomal doxorubicin has been recognized as an effective treatment in patients with refractory solid tumors. To our knowledge, this is the first case of relapsed Hodgkin lymphoma with breast involvement successfully treated with this association. PMID- 16313266 TI - Recurrent chemotherapy-induced tumor lysis syndrome (TLS) with renal failure in a patient with chronic lymphocytic leukemia - successful treatment and prevention of TLS with low-dose rasburicase. AB - INTRODUCTION: Rasburicase is a recombinant urate oxidase that is produced by a genetically modified Saccharomyces cerevisiae and has been approved for prophylaxis and treatment of tumor lysis syndrome in 2001. In several studies, rasburicase, given at a dose of 0.15-0.2 mg/kg for up to 7 d, proved to be highly effective in lowering urate levels. CASE REPORT: We report the case of a patient with chronic lymphatic leukemia (CLL) who experienced tumor lysis syndrome (TLS) with acute renal failure after fludarabine/cyclophosphamide chemotherapy and after bendamustine treatment. During the first episode of TLS, after fludarabine/cyclophosphamide (creatinine 3.3 mg/dL, urate 24.6 mg/dL), the patient received rasburicase 0.2 mg/kg for 3 d. Urate levels decreased below the lower limit of normal and renal function recovered. After bendamustine therapy, given for disease progression 8 months later, TLS with acute oliguric renal failure re-occurred (creatinine 3.1 mg/dL, urate 20.8 mg/dL). The patient was treated with hyperhydration and two doses of rasburicase (0.056 mg/kg), resulting in a prompt decrease of the urate level and recovery of renal function. Both episodes of TLS were successfully treated with rasburicase in a lower dose than recommended by the manufacturer. During a second bendamustine course, TLS was successfully treated by low doses of rasburicase (0.056 mg/kg for 2 d). CONCLUSION: This is the first report of TLS in CLL after bendamustine chemotherapy reported in the literature. Treatment and prevention of TLS by low doses of rasburicase is possible and cost-effective. PMID- 16313267 TI - Hemosiderin deposits in chronic graft-vs.-host disease related myopathy. AB - Chronic graft-vs.-host disease (cGVHD) occurs in 20-50% of patients who survive for at least 100 d after allogeneic stem cell transplantation (SCT). cGVHD includes scleroderma-like skin changes, chronic cholangitis, obstructive lung disease and general wasting syndrome. Polymyositis or myopathy are rare manifestations of cGVHD with approximately 40 reported cases. Polymyositis accompanied by hemosiderin deposits in cGVHD has been reported only once, and there are no reports on lipofuscin deposits in skeletal muscle cells in cGVHD. We report here on a 56-yr-old male who underwent allogeneic SCT in 1999 for osteomyelofibrosis and progressive hematopoietic insufficiency. In February 2004, the patient was hospitalized for progressive muscular weakness with loss of the ability to walk. Laboratory tests demonstrated normal values for serum creatine kinase, aldolase and lactic dehydrogenase; the ferritin level was highly elevated. The femoral muscle biopsy showed mostly perifascicular atrophy as well as numerous subsarcolemmal hemosiderin and lipofuscin deposits. Intravenous administration of the chelating agent deferoxamine was ineffective. Three weeks later the patient died of aspiration pneumonia. Interestingly, autopsy disclosed moderate hemosiderin deposits in the liver, the organ usually involved in hemosiderosis. PMID- 16313268 TI - Persistent panhypogammaglobulinemia with selected loss of memory B cells and impaired isotype expression after rituximab therapy for post-transplant EBV associated autoimmune hemolytic anemia. PMID- 16313269 TI - Polymorphisms of glutathione S-transferase mu1 (GSTM1) and theta 1 (GSTT1) genes in chronic myeloid leukaemia. PMID- 16313271 TI - Quality-adjusted life years: how useful in medico economic studies. AB - Cost-effectiveness analysis has evolved as a practical response to the need to allocate limited resources for health care. It can be used to compare interventions whose effects on health are different if the measure of effectiveness captures all the important health dimensions of the effects of the interventions. Using the quality-adjusted life year (QALY) as the unit of effectiveness attempts to approach this ideal and is currently the approach recommended by many consensus groups. Conventional QALYs represent time spend in a series of "quality-weighted" health states, where the quality weights reflect the desirability of living in the state. Many challenges arise when preferences are incorporated into an economic analysis. The purpose of this paper is to highlight some of the issues surrounding the use of QALYs and to encourage researchers to present their methodology in a clear and transparent way. PMID- 16313272 TI - Some issues with composite endpoints in clinical trials. AB - This article discusses some important issues that may arise in the current usage of composite endpoints as primary endpoints for demonstrating the efficacy of new drugs in clinical trials. The discussion focuses on time-to-event composite endpoints. Issues discussed include validity of a composite endpoint, the often lack of follow-up of patients beyond first event, the analysis of a composite endpoint, its sub-composite and individual component endpoints and their interpretation. Actual published examples in the literature are used to illustrate some of these problems. It is recommended that a clinical trial using a composite endpoint as the primary endpoint should be designed to include patient follow-up beyond the first event if possible. For data collected from such trials, basic formats for tabular presentation of trial data and for results of analysis of the composite endpoint, its sub-composite and individual component endpoints are proposed for transparency and ease of interpretation. PMID- 16313273 TI - Counterfeiting of medicinal drugs: issues and threats. AB - The increasing counterfeiting of medicinal drugs is a very worrying phenomenon, especially in developing countries. The risk for public health is quite real, particularly as these products are often sold out of any controlled distribution channel. PMID- 16313274 TI - The influence of weightlessness on pharmacokinetics. AB - The primary hostile factor during a spaceflight is the lack of gravity, which can induce space motion sickness and act on bones, muscles and the cardiovascular system. These physiological effects may modify the pharmacokinetics of the drugs administered during the flight producing reduced pharmacological activity or appearance of adverse effects. Given the small number of spaceflights and the difficulties of conducting experiments during missions, pharmacokinetic data obtained in flight are insufficient to determine if drug monitoring is necessary for the drugs present in the onboard medical kit. Therefore, validated earthbound models like tail-suspension performed with animals and long-term bedrest performed with human volunteers are used to simulate weightlessness and to study the pharmacokinetic variations of either absorption, distribution, or elimination of drugs. As a result of these studies, it is possible to make some dosing recommendations but more information is necessary to predict with precision all of the pharmacokinetic variations occurring in spaceflight. To collect more pharmacokinetic information, head-down bedrest studies are still the best solution and as saliva is an appropriate substitution for plasma for some drugs, salivary sampling can be planned during flights. PMID- 16313275 TI - An overview of the pharmacology and clinical efficacy of indapamide sustained release. AB - The relationship between blood pressure (BP) and cardiovascular risk is clearly established; hypertension increases the rate of cardiovascular. High systolic blood pressure (SBP) may be the main parameter involved in cardiovascular morbidity and mortality. The benefit of lowering BP, particularly with diuretics has been proven in many outcome studies. Indapamide, a thiazide-type diuretic, was available for many years at a dosage of 2.5 mg in an immediate release formulation. A new sustained release (SR) formulation has been developed in order to allow the same antihypertensive efficacy with a better acceptability profile. This paper reviews the pharmacology of indapamide 1.5 mg SR from the bench to the bedside. Indapamide has a dual mechanism of action: diuretic effect at the level of the distal tubule in the kidney and a direct vascular effect, both of which contribute to the antihypertensive efficacy of the drug. The SR formulation contains a hydrophilic matrix, which delivers a smoother pharmacokinetic profile. This avoids unnecessary plasma peak concentrations, which may be associated with side effects. Indapamide SR has now been extensively used in hypertensive patients, including those at increased risk, for example elderly or diabetic patients. It has been shown to decrease BP, particularly SBP, with 24-h efficacy, allowing a once-daily dosage. Studies have demonstrated BP lowering to be at least as effective as all major therapeutic classes including the more recent antihypertensive drugs. Beyond BP decrease, indapamide SR has also been shown to protect against hypertensive target-organ damage in the heart and the kidney and to have a favorable metabolic profile. A broad evidence-base has accumulated to support the benefit of indapamide 1.5 mg SR in hypertensive patients, alone or as part of combination therapy, as recommended by the majority of guidelines. PMID- 16313276 TI - Pharmacological interventions that directly stimulate or modulate insulin secretion from pancreatic beta-cell: implications for the treatment of type 2 diabetes. AB - Blood glucose concentration is controlled by a number of hormone and neurotransmitter signals, either increasing or reducing glucose levels in the case of hypoglycemia or hyperglycemia, respectively. The pancreatic beta-cell responds to an increase in circulating glucose levels by a cascade of metabolic and electrophysiological events leading to the secretion of insulin. Type 2 diabetes is a metabolic disorder characterized by chronic hyperglycemia; the progressive pancreatic beta-cell dysfunction, with altered insulin production and secretion, is a major pathophysiological determinant of the disease together with the resistance of insulin-sensitive tissues to the action of the hormone. Hence, drugs which stimulate or enhance insulin secretion will reduce plasma glucose concentrations; this lowering of hyperglycemia will, in turn, reduce the occurrence of long-term complications. K(ATP) channels play a critical role in insulin secretion and can be considered as transducers of glucose-induced metabolic changes into biophysical events leading to the exocytosis of insulin granules. All currently marketed insulin secretagogues, sulfonylureas and glinides, target the beta-cell K(ATP) channels and reduce their opening probability. They induce insulin release regardless of the plasma glucose concentration, thus favoring the occurrence of hypoglycemia in the fasting state. Despite the intensive use of current drugs, many patients suffering from type 2 diabetes still exhibit poor glycemic control, others fail to respond to the treatment, and some develop serious complications. Therefore, there is a real need for innovative compounds, either enhancing insulin secretion from the pancreas or improving insulin action on the hormone-sensitive tissues. Here, we overview the existing and novel approaches targeting the beta-cell to enhance the release of insulin, with special emphasis on new ways of amplifying insulin secretion in a glucose-dependent manner. PMID- 16313277 TI - Lack of hypo/hyperglycemic effects of reboxetine in diabetic and non-diabetic rats. AB - Depression occurs frequently with diabetes affecting the quality of life. All major classes of antidepressants have been shown to have a direct pharmacologic effect on metabolic function, which further worsens glycemic control. There were no reports of effects of reboxetine on glucose and insulin levels. We observed that 21 days administration of reboxetine (2.5 mg/kg, i.p.) in streptozotocin (55 mg/kg, i.p.) induced diabetic and non-diabetic rats produced no significant alteration in glucose levels in diabetic and non-diabetic rats. Serum insulin levels were also not significantly changed by reboxetine (2.5 mg/kg, i.p.). Selective inhibition of noradrenaline reuptake by reboxetine has been suggested to be responsible for lack of effects on glucose and insulin levels. PMID- 16313278 TI - Oxidative stress activates MMP-2 in cultured human coronary smooth muscle cells. AB - Oxidative stress is a cardinal feature of the inflammatory process and is involved in various pathologies including atherosclerosis. One of the important mechanisms in which oxidative stress may play a role is activation of matrix metalloproteinases such as MMP-2, which are involved in plaque destabilization. We investigated the mechanisms by which oxidative stress induces MMP-2 activation in cultured human coronary artery smooth muscle cells. Using zymography and Western blot analysis, we showed that oxidized low-density lipoproteins activate MMP-2 through up-regulation of the expression and activation of a membrane-type 1 matrix metalloproteinase (MT1-MMP). A second mechanism of MMP-2 activation involves oxidative radicals generated by the xanthine/xanthine oxidase complex (X/Xo). Research on these two mechanisms of MMP activation could lead to the elaboration of new vascular therapies for the treatment of atheroma based on interruption of a specific oxidative stress pathway. PMID- 16313279 TI - Lamotrigine pharmacokinetic/pharmacodynamic modelling in rats. AB - The aim of this study was to perform a pharmacokinetic/pharmacodynamic (PK/PD) modelling of lamotrigine following its acute administration to rats. Adult male Wistar rats were given 10 mg/kg of lamotrigine intraperitoneally. Plasma and brain samples were obtained at predetermined times over 120 h post-dose and analysed by liquid chromatography. The anticonvulsant profile against maximal electroshock seizure stimulation was determined over 48 h after dosing. As a linear relationship between lamotrigine plasma and brain profiles was observed, only the plasma data set was used to establish the PK/PD relationship. To fit the effect-time course of lamotrigine, the PK/PD simultaneous fitting link model was used: the pharmacokinetic parameters and dosing information were used in the one compartment first-order model to predict concentrations, which were then used to model the pharmacodynamic data with the sigmoid Emax model, in order to estimate all the parameters simultaneously. The following parameters were obtained: Vd = 2.00 L/kg, k(abs) = 8.50 h(-1), k(el) = 0.025 h(-1), k(e0) = 3.75 h(-1), Emax = 100.0% (fixed), EC50 = 3.44 mg/L and gamma = 8.64. From these results, it can be stated that lamotrigine is extensively distributed through the body, its plasma elimination half-life is around 28 h and a lamotrigine plasma concentration of 3.44 mg/L is enough to protect 50% of the animals. When compared with humans, the plasma concentrations achieved with this dose were within the therapeutic concentration range that had been proposed for epileptic patients. With the present PK/PD modelling it was possible to fit simultaneously the time-courses of the plasma levels and the anticonvulsant effect of lamotrigine, providing information not only about the pharmacokinetics of lamotrigine in the rat but also about its anticonvulsant response over time. As this approach can be easily applied to other drugs, it becomes a useful tool for an explanatory comparison between lamotrigine and other antiepileptic drugs. PMID- 16313280 TI - Inhibition of angiotensin II- and endothelin-1-stimulated proliferation by selective MEK inhibitor in cultured rabbit gingival fibroblastsdagger. AB - We investigated the implication of extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) in the proliferation stimulated by angiotensin II (Ang II) and endothelin-1 (ET-1) in cultured rabbit gingival fibroblasts (CRGF). Ang II stimulated activation of ERK1/2 and the activation was inhibited by CV-11974, an AT1 antagonist, and saralasin, an AT1/AT2 antagonist, but not by PD123,319, an AT2 antagonist in the CRGF. Ang II-stimulated proliferation was inhibited by PD98059 or U0126, selective MEK inhibitors. Furthermore, ET-1 stimulated proliferation via G-protein-coupled ETA receptors, which were identified by Western blot analysis of membrane protein from the CRGF. ET-1 also stimulated activation of ERK1/2 and the activation was inhibited by BQ-123, an ETA inhibitor, and TAK044, an ETA/ETB inhibitor, but not by BQ-788, an ETB inhibitor. ET-1-stimulated proliferation was inhibited by PD98059 or U0126. These findings suggest that ERK1/2 play a role in the signaling process leading to proliferation stimulated by Ang II and ET-1 via G-protein-coupled receptors, AT1 and ETA in CRGF. PMID- 16313281 TI - Chemopreventive efficacy and anti-lipid peroxidative potential of Jasminum grandiflorum Linn. on 7,12-dimethylbenz(a)anthracene-induced rat mammary carcinogenesis. AB - The aim of this study was to investigate the chemopreventive efficacy and anti lipid peroxidative potential of Jasminum grandiflorum Linn. on 7,12 dimethylbenz(a)anthracene (DMBA)-induced rat mammary carcinogenesis. Mammary tumors were developed by a single subcutaneous injection of 25 mg DMBA in 1 mL emulsion of sunflower oil and physiological saline. The tumor incidence and tumor volume that formed in the breast were determined. Oral administration of ethanolic extract of J. grandiflorum flowers (JgEt) at a dose of 300 mg/kg body weight for 14 weeks to DMBA-injected animals completely prevented the formation of tumors in the pre-initiation period. JgEt also exerted significant anti-lipid peroxidative effect and improved the antioxidant defense system in DMBA-treated rats. The results of this study clearly indicate that JgEt has potent chemopreventive efficacy in experimental mammary carcinogenesis and further studies are warranted to isolate and characterize the bioactive principle from JgEt. PMID- 16313282 TI - Antispasmodic and bronchodilator activities of St John's wort are putatively mediated through dual inhibition of calcium influx and phosphodiesterase. AB - The crude extract of aerial parts of St John's wort (Hypericum perforatum) (Hp.Cr) and its fractions were studied in vitro for its possible spasmolytic and bronchodilator activities to rationalize some of its medicinal uses. In rabbit jejunum preparations, Hp.Cr caused a concentration-dependent relaxation of both spontaneous and K+ (80 mm)-induced contractions at a similar concentration range (0.1-1.0 mg/mL), similar to that produced by papaverine, whereas verapamil was relatively potent against K+-induced contractions. Hp.Cr shifted the Ca2+ concentration-response curves (CRCs) to the right, similar to that caused by papaverine or verapamil and also caused leftward shift of isoprenaline-induced inhibitory CRCs, similar to papaverine. In guinea-pig tracheal preparations, Hp.Cr caused relaxation of carbachol and K+-induced contractions at similar concentrations (0.01-0.3 mg/mL) and also shifted the isoprenaline-induced inhibitory CRCs to the left, similar to that caused by papaverine. In rabbit aorta preparations at rest, Hp.Cr produced a moderate vasoconstriction, while exhibited vasodilator effect against phenylephrine and K+-induced contractions. Papaverine and verapamil also produced similar non-specific vasodilation, but were devoid of any vasoconstrictor effect. Hp.Cr caused suppression of atrial force of contractions at concentrations about 20 times higher than those that produced inhibitory effect in smooth muscle preparations, similar to papaverine. These results suggest that the spasmolytic effects of Hp.Cr are mediated through dual inhibition of calcium influx and phosphodiesterase (PDE)-like mechanisms, which might explain the medicinal use of St John's wort in the disorders of gastrointestinal and respiratory tracts. Furthermore, the presence of Ca2+ antagonist and PDE inhibitory-like constituents might also be contributing to some extent in the well established use of plant in depression. PMID- 16313283 TI - Comparative trial of tramadol/paracetamol and codeine/paracetamol combination tablets on the vigilance of healthy volunteers. AB - Combination of tramadol 37.5 mg/paracetamol 325 mg (a), or codeine 30 mg/paracetamol 500 mg (b) or 300 mg have similar pain efficacy but a difference has been suggested concerning their adverse events on vigilance. In clinical practice, combinations are usually given at the above-mentioned dosage three to four times a day. The aim of this study was to compare a single dose of these two combinations (a) and (b) in 24 healthy young volunteers on visual choice reaction time (CRT, ms). Results show a longer CRT (P < 0.05) (up to 4% of the control value 3 h post-dosing) and a higher report of somnolence in the codeine/paracetamol group compared with tramadol/paracetamol group (50% vs. 4% of the subjects). This observation is important and proves that even a single dosage of these largely used drugs may have a significant effect. This finding should be further investigated in elderly subjects who consume largely these drugs for chronic pain alleviation and who are more prone to this kind of adverse event. PMID- 16313284 TI - Long-term complications of sulphur mustard poisoning in severely intoxicated Iranian veterans. AB - Sulphur mustard (SM) is an alkylating chemical warfare agent that was widely used during the World War I and in the Iran-Iraq conflict. Delayed complications of SM in different organs and their severity correlations have not previously been reported. Dermatological, ophthalmological, neurological and respiratory examinations, as well as spirometry, gasometry, high resolution computed tomography scanning of the chest, electromyography, nerve conduction velocity, cell blood counts, flow-cytometric analyses, and measurement of serum immunoglobulins and complement factors were performed on all severely SM poisoned veterans in the province of Khorasan, Iran. Haematological and immunological studies were also performed on a control group consisting of 35 healthy male subjects. The severity of dermal, ocular and respiratory complications were classified into four grades and their correlations with each other as well as with the haematological and immunological parameters were determined, using Spearman's rank correlation test. Forty male patients (aged 43.8 +/- 9.8 years) with confirmed SM poisoning 16-20 years after the exposure were studied. The most common complications were found in the lungs (95%), peripheral nerves (77.5%), skin (75%), and eyes (65%). WBC, RBC, haematocrit (HCT), IgM, C3, and the percentages of monocytes and CD3+ lymphocytes were significantly (P < 0.042) higher and the percentage of CD16 + 56 positive cells was significantly (P = 0.006) lower in the patients than in the control group. The severity of respiratory complications revealed a significant correlation with the severity of ocular complications (r = 0.322, P = 0.043), as well as with the haemoglobin (r = 0.369, P = 0.024) and HCT (r = 0.470, P =0.003). Although late complications of SM poisoning in the skin, eyes, and respiratory system are mainly due to its direct toxic effects, the neuromuscular, haematological and immunological complications are probably the result of systemic toxicity. PMID- 16313286 TI - Determinants of response to first HAART regimen in antiretroviral-naive patients with an estimated time since HIV seroconversion. AB - OBJECTIVE: To study the determinants of immunological and virological response to highly active antiretroviral therapy (HAART) in naive patients, adjusting for time since HIV-1 seroconversion. DESIGN: Data from HIV-cohort studies where dates of seroconversion have been reliably estimated. Methods In previously untreated patients, short- and long-term marker responses from HAART initiation (three or more antiretroviral drugs) to the end of follow-up or any treatment modification were considered using mixed effects models accounting for undetectable HIV viral load and informative dropout. RESULTS: In total, 943 patients were treated with a first HAART regimen for a median of 29 months. In adjusted analyses, compared with a reference group of homosexual men without AIDS initiating treatment 4 years after seroconversion, injecting drug users (IDUs) were treated at similar CD4 and HIV RNA levels but had poorer short-term virological response (2.54 vs 2.13 log(10) HIV-1 RNA copies/mL at 1.5 months, P=0.03) and poorer long-term immunological response (522 vs 631 cells/microL at 24 months, P<0.0001). Although individuals with AIDS at HAART initiation had lower CD4 counts (206 vs 382 cells/microL, P<0.0001), their immunological responses were similar to those of individuals without AIDS. Similarly, individuals further from seroconversion started HAART at lower CD4 counts (e.g. 311 vs 382 cells/microL at vs before 9 years from seroconversion, P<0.0001), but had similar CD4 responses. However, they experienced poorer long-term virological response (0.67 log(10) copies/mL/year smaller decline, P<0.0001) compared to those treated before 9 years from seroconversion. CONCLUSION: Taking into account the time elapsed since seroconversion, this study suggests that careful choices of initial treatment should be made and intensive follow-up carried out in high-risk subgroups such as IDUs who have poorer responses. PMID- 16313287 TI - Impact of highly active antiretroviral therapy on blood pressure in HIV-infected patients. A prospective study in a cohort of naive patients. AB - OBJECTIVES: To assess the impact of highly active antiretroviral therapy (HAART) on the blood pressure (BP) of naive patients after 1 year of treatment. METHODS: A prospective, observational study of 95 HIV-positive patients in our Unit starting HAART between January 2001 and October 2002 and maintaining the same regimen for 48 weeks of follow-up was carried out. Data on blood pressure (BP) and demographic, epidemiological, clinical, immunovirological and therapeutic characteristics related to HIV infection were collected prior to HAART and at week 48. High blood pressure (HBP) [systolic BP (SBP) > or =140 mm Hg and/or diastolic BP (DBP) > or =90 mm Hg] was defined according to international criteria. RESULTS: Of the 95 patients, 78 were men, 44% had AIDS and 68% were smokers, and their mean age was 40 years. At week 48 the prevalence of HBP was 26% and SBP, DBP and pulse pressure (PP) increased (121.8 versus 116.6 mm Hg, P=0.0001; 76.3 versus 69.7 mm Hg, P=0.004; 46.9 versus 43.8 mm Hg, P=0.001, respectively). Univariate analysis showed that HBP was associated with older age, higher body mass index (BMI), higher baseline lipids, and higher baseline BP. A linear regression model adjusting for age and sex suggested a significant impact of older age, higher baseline SBP, higher baseline hypercholesterolaemia and lower baseline CD4-cell count on SBP increase. CONCLUSIONS: Blood pressure increased after 48 weeks of HAART, leading to an important prevalence of hypertension. The increase in SBP depended on age and baseline lipid profile and immunological status. BP should be periodically measured and treated when necessary in HIV-infected patients on HAART. PMID- 16313288 TI - Effects of highly active antiretroviral therapy on paediatric metabolite levels. AB - OBJECTIVES: Highly active antiretroviral therapy (HAART) has extended survival of HIV-infected children into adulthood, raising concerns about long-term metabolic changes in childhood. METHODS: A longitudinal study of metabolite levels in paediatric HIV-infected patients before and after starting HAART (January 2000 to June 2003). The effects of HAART on nonfasting blood levels of total (TC), high density lipoprotein (HDL), and low-density lipoprotein (LDL) cholesterol, cholesterol ratio and lactate were analysed using mixed-effects regression. RESULTS: A total of 146 children attended 1208 appointments (median 6.7/child). Of these, 99 (68%) were African. At baseline, 75 (51%) were on HAART and had higher TC (4.19 vs 3.49 mmol/L, P<0.0001), HDL (1.03 vs 0.82 mmol/L, P<0.0001), and LDL (2.54 vs 2.11 mmol/L, P=0.0003) than those not on HAART. Metabolites increased with time on HAART exposure and then stabilized. At 2 years, TC had increased by 0.93 mmol/L (P<0.0001), with 29 children (20%) having repeated TC levels above the 95th centile. LDL and HDL had increased by 0.69 and 0.31 mmol/L at 2 years, respectively (both P<0.0001). Lactates declined with increasing age ( 0.06 mmol/L/year, P=0.0001). CONCLUSIONS: This is the first cohort study to demonstrate significant elevations of HDL as well as LDL in children on HAART. This rise in cardio-protective HDL may represent a positive effect of treatment. PMID- 16313289 TI - Effect of GB virus C on response to antiretroviral therapy in HIV-infected Brazilians. AB - OBJECTIVES: GB virus C (GBV-C) infection is associated with delayed mortality in HIV-infected people in most, but not all, studies. Previous investigations of the effect of GBV-C viraemia on response to antiretroviral therapy (ART) were inconclusive. To determine the effect of GBV-C on ART, we retrospectively analysed plasma samples taken from patients in a prospective randomized clinical trial of ART in HIV-positive Brazilians. METHODS: GBV-C viraemia was characterized by testing stored serum samples from 175 participants by reverse transcriptase-polymerase chain reaction (RT-PCR). Subjects were randomized to receive indinavir (n=59), zidovudine and lamivudine (n=58), or zidovudine, lamivudine and indinavir (n=58). The effect of GBV-C viraemia on the average change in HIV viral load and CD4 count following initiation of therapy was evaluated in a multiple regression analysis. RESULTS: The prevalence of GBV-C viraemia was similar to that observed in previous studies (24%). HIV viral load decreased following ART to a significantly greater extent in patients with GBV-C viraemia (by 0.48 log(10) HIV-1 RNA copies/mL, P=0.009, adjusting for age, ART group, and baseline CD4 count). Although there was no significant difference in change in CD4 count between individuals with and without GBV-C viraemia overall, CD4 counts were higher following 48 weeks of therapy in GBV-C viraemic individuals receiving the least potent ART regimen (zidovudine and lamivudine) compared with those without GBV-C infection. CONCLUSIONS: GBV-C viraemia is associated with an enhanced reduction of HIV viral load in response to ART. In this study of treatment-naive individuals during 48 weeks of follow up, patients with GBV-C viraemia had reductions in HIV viral load that were approximately 0.5 log copies/mL greater than those found in patients without GBV-C viraemia. This is similar to reductions observed with nucleoside reverse transcriptase inhibitors. PMID- 16313290 TI - Clinical outcomes of first antiretroviral regimen in HIV/hepatitis C virus co infection. AB - OBJECTIVES: Despite the benefits of HAART, initiation of antiretroviral therapy in HIV-HCV co-infected patients is often delayed as a consequence of patient and physician concern pertaining to liver toxicity. It is unclear whether this is justified. METHODS: We retrospectively evaluated treatment duration and outcome in 186 patients initiating a first HAART regimen. RESULTS: Despite frequent HIV RNA suppression and CD4 T-cell increase following initiation of HAART, the median duration of therapy was only 8 months. Therapy was discontinued primarily for gastrointestinal intolerance (26%), poor adherence (19%), neurocognitive side effects (13%), and substance abuse (6%). Regimes were changed to reduce pill burden and/or frequency of dosing as well (11%). Only six (4%) subjects interrupted therapy as a result of clinically apparent liver toxicity. None were on low dose ritonavir-containing therapy. In those subjects remaining on HAART for at least 12 months, the median ALT level increased marginally from a baseline of 44 IU/mL to 56 IU/mL. The median AST was 44 IU/mL at baseline and at month 12. CONCLUSIONS: These results support our contention that regimen potency, durability, and extrahepatic side effect profile should remain the paramount considerations related to the selection of HAART regimen in HIV-HCV co-infection. PMID- 16313291 TI - Pronounced lipoatrophy in HIV-infected men receiving HAART for more than 6 years compared with the background population. AB - OBJECTIVES: To establish the prevalence and quantify the severity of body fat redistribution and dyslipidaemia in HIV-infected men after long-term highly active antiretroviral therapy (HAART) compared with the background population. METHODS: In a cross-sectional study, we included 87 HIV-infected men who had received HAART for at least 6 years and 34 HIV-negative men. Regional body composition was assessed using dual-energy X-ray absorptiometry. Fasting metabolic parameters were obtained. Associations between regional body fat distribution and metabolic parameters were evaluated. RESULTS: HIV-infected patients and controls did not differ with regard to height and lean body mass. Compared with controls, HIV-infected men had reduced total fat mass (median 12.3 versus 19.2 kg, P<0.001), limb fat mass (4.3 versus 7.9 kg, P<0.001), and trunk fat mass (6.7 versus 10.8 kg, P<0.001) and higher trunk/limb fat ratio (1.7 versus 1.2, P<0.001). Also, patients without clinical lipodystrophy had reduced amounts of limb and trunk fat. In HIV-infected men, triglyceride levels were higher (2.0 versus 1.2 mmol/L, P<0.001), high-density lipoprotein (HDL) cholesterol levels were lower (1.2 versus 1.3 mmol/L, P<0.05) and insulin levels were higher (40.8 versus 29.9 pmol/L, P<0.01) than in controls. All adverse metabolic parameters correlated with increased trunk/limb fat ratio, and insulin levels correlated positively with trunk fat mass (P<0.01). CONCLUSION: Peripheral as well as central fat loss is a general characteristic of HIV-infected men after long-term HAART. Although lipoatrophy was the dominant morphological presentation, the adverse metabolic parameters were mainly associated with the increased ratio of trunk/limb fat. PMID- 16313292 TI - Cervical screening in HIV-positive women: characteristics of those who default and attitudes towards screening. AB - OBJECTIVES: To determine the characteristics of HIV-positive women who undergo cervical screening and to identify negative attitudes and experiences of these women to screening and the factors associated with these. METHODS: We compared the Royal Free Cohort data from 59 newly diagnosed HIV-infected women, 31 of whom did and 28 of whom did not attend for cervical screening in 2001, and from 227 women under active cervical screening follow-up (at least one cervical screen since June 2001) and 88 HIV-infected women lost to follow-up (not screened since January 2001). Attitudes to screening were investigated with the aid of a questionnaire given to all women attending clinic who had had a previous colposcopy. RESULTS: Of the 59 newly HIV diagnosed women, 31 (53%) underwent cervical screening. These 31 women were more likely to be heterosexual (100 vs 89%, P=0.05), to have lower median viral loads (< 50 vs 3210 HIV-1 RNA copies/mL) and to be receiving antiretrovirals (ARVs) (74 vs 54%, P=0.1) than those not screened. Of the 315 women who had at least one screen, 72% returned for further follow-up. There were no differences in age or ethnicity between these groups. Those under active follow-up had a higher CD4 count (P=0.04) and lower viral load (P=0.0001) at their last visit. They were also more likely to be on highly active antiretroviral therapy (HAART) (68 vs 52%, P=0.006). A total of 78 of 104 questionnaires (75%) were returned. Women participating in the questionnaire study were mainly of back ethnicity (68%), did not speak English as their first language (59%) and were taking ARVs (76%). Most agreed that regular smears and colposcopy were valuable. Women of white ethnicity, and those speaking English as a first language, were more likely to dislike colposcopy compared with those of nonwhite ethnicity (87 vs 25%, respectively, P=0.0007) and not speaking English as a first language (74 vs 26%, respectively, P=0.002). Those of white ethnicity were more likely to find smears and colposcopy painful (60 and 73%) compared with those of black ethnicity (46 and 51%, P=0.47 and 0.28, respectively). CONCLUSIONS: Our results suggest that women on HAART with better disease control, older women, and those of black African ethnicity are more likely to take up cervical screening. Cervical screening experience varies by ethnicity and language. PMID- 16313293 TI - Depletion of mitochondrial DNA copies/cell in peripheral blood mononuclear cells in HIV-1-infected treatment-naive patients. AB - OBJECTIVES: Mitochondrial toxicity is believed to be the main reason for adverse effects related to nucleoside reverse transcriptase inhibitors (NRTIs). The aim of the present study was to compare mitochondrial toxicity in NRTI-treated HIV positive patients, HIV-positive treatment-naive patients and HIV-negative controls by comparing mitochondrial DNA (mtDNA) copies/cell in peripheral blood mononuclear cells (PBMCs) and lactate/pyruvate (L/P) ratios in the different groups. METHODS: We enrolled 60 participants in the study: 31 patients on combined antiretroviral therapy (CART), 14 HIV-positive treatment-naive patients and 15 HIV-negative controls. mtDNA (copies/cell) in peripheral blood was analysed using quantitative real-time polymerase chain reaction (PCR). Standard curves and serial dilutions of plasmid-cloned mitochondrion and retinoblastoma (RB1) PCR products with known concentrations were generated to estimate the mtDNA and nuclear DNA (nDNA) copy numbers in each sample. The L/P ratio was enzymatically and spectrophotometrically analysed in samples from individuals in a fasted, non-exercise state. Results The median mtDNA copy number was 63 copies/cell (interquartile range 33-94) in HIV-positive patients and 153 (132 283) in HIV-negative controls (P<0.001). No significant difference was seen between the HIV-positive NRTI-exposed patients and the HIV-positive treatment naive patients. Current use of didanosine was negatively correlated with depletion of mtDNA (r=-0.36, P=0.046). HIV-positive patients also had a higher L/P ratio compared with HIV-negative controls (P=0.004). CONCLUSIONS: The number of mtDNA copies/cell in PBMCs was depleted in HIV-positive treatment-naive patients as well as in HIV-positive NRTI-exposed patients. HIV-positive patients also had a higher L/P ratio compared with HIV-negative controls, which supports this conclusion. The study suggests that neither mtDNA in PBMCs nor L/P ratio is a good marker of NRTI-associated mitochondrial toxicity. PMID- 16313294 TI - A recent increase in AIDS at Lyon University Hospitals: patient characteristics and comparisons with previous years. AB - BACKGROUND: A 36% increase in the incidence of AIDS was observed in 2002/2003 compared with 2000/2001 at Lyon University Hospitals. OBJECTIVES: We compared the characteristics of these patients with the characteristics of those diagnosed previously with AIDS. METHODS: Data for all patients with AIDS diagnosed at Lyon University Hospitals were analyzed. The data were collected prospectively. Multiple logistic regression was used for analysis. RESULTS: The variables independently associated with an AIDS diagnosis in 2002/2003 compared with the 1985-1989 period were: homosexual exposure [odds ratio (OR) 0.4; 95% confidence interval (CI) 0.2-0.8]; heterosexual exposure in an endemic area (OR 22.5; 95% CI 6.8-74.8), compared with other exposure to HIV; lymphoma as initial AIDS event (OR 10.3; 95% CI 2.7-39.1) compared with Pneumocystis carinii pneumonia; and age at first AIDS event aged 34-38 years (OR 2.5; 95% CI 1.0-6.4), aged 39-46 years (OR 5.1; 95% CI 2.2-11.8), and aged 47-84 years (OR 10.6; 95% CI 4.5-25.1) compared with aged <30 years. The variables independently associated with an AIDS diagnosis in 2002/2003 compared with the 1997/2001 period were age at first AIDS event aged 34-38 years (OR 0.4; 95% CI 0.2-0.9) compared with aged <30 years. CONCLUSION: Recently diagnosed AIDS patients differed from those diagnosed previously, showing an epidemic switch in different populations. The characteristics of the AIDS population in 2002/2003 might reflect public health messages disseminated around 10 years ago or more for the prevention of HIV transmission. Anticipation of populations affected by the AIDS epidemic is difficult. PMID- 16313295 TI - Antiviral efficacy and resistance in patients on antiretroviral therapy in Kigali, Rwanda: the real-life situation in 2002. AB - Our study aimed to complete the published data on ARV therapy in Africa by describing the baseline situation in Rwanda before the launch of a large ARV programme (ESTHER). Prescription habits, frequency and reasons for treatment interruptions but also antiviral efficay, resistance to ARVs and genotypic variability of the viruses present in Rwanda were analysed. Among the 233 patients included in the study, it appeared that a vast majority (91%) were under triple therapy and that half of them had experienced at least one treatment interruption caused mainly by drug shortage or financial difficulties. Among 60 blood samples analysed, 26 were in virological failure with a viral load above 1000 RNA copies/ml and 11 presented major drug resistance mutations. Finally, virological failure could mainly be explained by the high frequency of treatment interruptions but also by the emergence of drug resistance mutations. Consequently the major objective for the ESTHER programme to improve the situation in Rwanda will be to reduce the drug shortage and facilitate the financial accessibility of the treatments. PMID- 16313296 TI - HLA and eye disease: a synopsis. AB - Human leukocyte antigen (HLA) gene products have been implicated in the pathogenesis of an increasing number of eye diseases, mainly inflammatory in nature. This perspective reviews the current hypotheses for why HLA polymorphisms are associated with specific eye diseases. Statistical problems in studies involving HLA associations are discussed, and possible solutions outlined. The relevance of HLA testing in routine ophthalmic practice, its practical and cost implications is also assessed. PMID- 16313297 TI - Lack of association between pro-inflammatory cytokine (IL-6, IL-8 and TNF-alpha) gene polymorphisms and Graves' disease. AB - Graves' disease (GD) is a common, autoimmune disease involving the thyroid gland, and it has been previously suggested that pro-inflammatory cytokines are involved in the disease's pathogenesis. The aim of this study was to test whether the interleukin (IL)-6 gene promoter region, or tumour necrosis factor (TNF)-alpha or IL-8 gene 3'-untranslated region (3'-UTR) polymorphisms could provide useful genetic markers for an individual's susceptibility to GD. A normal control group of 60 healthy people and 95 patients featuring GD were examined. Polymerase chain reaction (PCR)-based restriction analysis was performed for the three gene polymorphisms using endonucleases BsrBI, NcoI and ApaLI, respectively. We found no significant difference between the frequencies of genotype and allelic variants for the IL-6 gene promoter (-572 G/C), the TNF-alpha gene promoter (-308 A/G) and the IL-8 gene 3'-UTR (2767 A/G) for GD patients and for normal controls. Cytokines are a large group of proteins that may elicit multiple effects upon immunological reactions. It still appears to be very worthwhile to continue to aggressively search for cytokine gene polymorphisms in order to predict the development of such disease. PMID- 16313298 TI - Interleukin-6 gene polymorphism is an age-dependent risk factor for myocardial infarction in men. AB - Several studies show that inflammatory components may contribute to atherosclerosis and increase the risk for myocardial infarction (MI). Interleukin 6 (IL-6) is a key pro-inflammatory and immune-modulatory cytokine of relevance for cardiovascular diseases. In this case-control study, 200 patients with MI and 257 healthy controls were genotyped for the polymorphism present in -174 promoter region of the IL-6 gene. Plasma concentrations of IL-6 and C-reactive protein (CRP) in a group of patients and controls were measured. The -174 C allele was associated with an increased risk of developing MI (OR = 2.886, c.i. = 1.801 4.624, P = 0.0001) in older patients, while no association was found in younger ones. The IL-6 plasma levels were higher in patients with MI carrying the CC genotype than in GG patients (CC carriers, IL-6 = 2.97 pg mL(-1) vs. GG carriers = 1.81 pg mL(-1), P = 0.016). A positive correlation of IL-6 levels with those of CRP in serum from patients with MI was also found. Data from this study suggest that the C allele of the promoter polymorphism in the IL-6 gene is a risk factor for MI in the elderly, and the production of the IL-6 is differentially affected by different genotypes of the IL-6 -174 promoter polymorphism. PMID- 16313299 TI - Duck (Anas platyrhynchos), Japanese quail (Coturnix coturnix japonica) and other avian interleukin-2 reveals significant conservation of gene organization, promoter elements and functional residues. AB - We compared the gene, promoter and cDNA sequences, and the predicted protein structure of duck and quail interleukin-2 (IL-2), a major immunomodulatory cytokine, with the known sequences of other avian and human IL-2. Analysis of the gene organization showed significant similarity with the overall organization of mammalian IL-2 genes, with four exons and three introns and a very short 5' untranslated regions. The second intron was the biggest in all the IL-2 sequences. The third intron was of similar size in chicken and quail, whereas in duck it was found to be slightly longer. Promoter sequence analysis of the IL-2 gene revealed remarkable conservation of the functionally important residues. The transcription factor binding sites such as those for AP-1, NF-AT, CD 28 RE and OCT, the TATA box and the predicted transcription start site with respect to chicken IL-2 sequence showed total conservation in duck, quail and turkey IL-2 promoters. Comparative analysis of the avian IL-2 cDNAs such as those of chicken, turkey, duck, quail, goose and Muscovy duck, revealed significant conservation of the nucleotide and predicted amino acid sequences. They showed nucleotide identity levels varying from 75% to 85%, amino acid level identity from 58% to 72% and amino acid similarity from 71% to 80% with each other. In the predicted protein secondary structure, the four essential alpha-helices and the hydrophobic amino acids in the heptad repeats forming the core structure of IL-2 molecules were conserved in all the avian and the human IL-2. The present study reveals high-level conservation of the gene; cDNA structure and regulatory elements of avian IL-2. This indicates highly conserved functions and probable functional cross-reactivity of this major immunomodulatory cytokine among birds. PMID- 16313300 TI - ICAM-1 polymorphisms and development of cutaneous malignant melanoma. AB - Tumour growth in cutaneous malignant melanoma (CMM) is mediated by cell adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1). ICAM-1 expression is associated with increasing Breslow thickness of vertical growth phase tumours and, in patients with stage 1 disease, may be associated with disease free and patient survival. In this study we have investigated whether two single nucleotide polymorphisms (SNPs) in the ICAM-1 gene encoding amino acid substitutions in codons 241 and 469 of the expressed ICAM-1 molecule are associated with susceptibility to and markers of prognosis (including tumour Breslow thickness) in CMM. A total of 164 CMM patients and 264 cancer-free controls were genotyped for these SNPs by the 5' nuclease assay for allelic discrimination (TaqMan). No genotypes showed any significant associations with CMM susceptibility, although there was a non-significant increase in frequency of the ICAM-1 469 AA genotype among CMM patients vs. controls (38.4% vs. 29.9%; P = 0.11). However, the ICAM-1 241 GG genotype was significantly decreased in frequency among patients with primary invasive tumours of greater Breslow thickness (72.5% vs. 91.2%; P = 0.013; OR = 0.25 (0.072-0.85)). These results provide no evidence for a role for the ICAM-1 codon 241 and 469 SNPs in determining susceptibility to CMM, but provide preliminary evidence that the role of ICAM-1 polymorphism in modulating tumour growth in CMM requires further investigation in a larger study group. PMID- 16313301 TI - Alternative splicing of IL-24 in melanocytes by deletion of exons 3 and 5. AB - Two novel interleukin-24 (IL-24) splice variants were identified in normal human melanocytes by sequencing cloned polymerase chain reaction (PCR) products that are not expressed in metastatic melanoma. These gene products have been generated by differential skipping of exons 3 (IL-24 delE3) and 5 (IL-24 delE5). IL-24 delE3 has limited sequence identity to the IL-24-interacting protein mda-7s, and IL-24 delE5 is homologous to IL-24. PMID- 16313302 TI - Novel polymorphism in the promoter region of the human nerve growth-factor gene. AB - We describe a novel T to C transition at position -198 from the transcription start of the human nerve growth-factor (NGF) gene. In British Caucasoid healthy control group that we have genotyped, T and C allele frequencies are 0.633 and 0.367, respectively. This polymorphism affects vitamin D receptor (VDR) binding to its motif in the NGF promoter. PMID- 16313303 TI - Polymorphisms in the interleukin-4 and IL-4 receptor genes and multiple sclerosis: a study in Spanish-Basque, Northern Irish and Belgian populations. AB - Cytokine gene polymorphisms are known to influence susceptibility and disease course of many autoimmune diseases. Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system white matter characterized by inflammation, demyelination and axonal damage. We analysed both the well-known intronic variable number of tandem repeat (VNTR) and +33 C/T single-nucleotide polymorphisms (SNP) in the IL-4 gene, as well as the functional Q551R SNP in the IL4-R gene in a cohort of three distinct populations comprising sporadic cases and controls from the northern Spanish Basque Country and Northern Ireland, as well as family trios from Belgium. The IL-4 +33 TT genotype was decreased in primary progressive (PP) versus relapsing-remitting (RR) patients in the Northern Irish population (OR = 0.14; 95% CI = 0.018-1.09). Two-marker haplotype distribution of the VNTR and +33 C/T SNP in PP patients differed from that seen in RR patients in Northern Ireland (P = 0.03). The R allele of the Q551R SNP was significantly under-transmitted in the Belgian trio families (P = 0.003), although this effect was not seen in the Northern Irish and Basque data sets. We did not identify IL-4-IL4-R gene-gene interaction in determining susceptibility or clinical parameters of MS. Disease or genetic heterogeneity or both may be responsible for the observed lack of reproduction in different populations. Our data reinforce recent findings for a role of IL4-R in susceptibility to MS. PMID- 16313304 TI - HLA-C locus alleles distribution in patients from northern Poland with psoriatic arthritis--preliminary report. AB - The aim of the study was to compare the frequency of human leucocyte antigen-C (HLA-C) locus alleles in patients with psoriatic arthritis and in healthy controls in the same ethnic group in Poland, and to correlate them with age of onset of psoriatic skin changes and joints symptoms. HLA-C locus alleles of 41 patients and 80 controls were determined by a polymerase chain reaction (PCR) low resolution method. The Cw*06 allele occurred more frequently (P adjusted for multiple comparison = 0.004) in patients with psoriatic arthritis than in controls. Patients who carried the HLA-Cw*06 allele had a significantly earlier mean age of onset of both psoriasis (P = 0.01) and arthritis (P = 0.008) compared with Cw*06-negative patients. Our results confirm the association between Cw*06 allele and psoriatic arthritis in the northern Poland population and suggest that the HLA-Cw*06 may determine not only the disease susceptibility, but also the age of onset of psoriatic arthritis. PMID- 16313305 TI - Polymorphisms at +49A/G and CT60 sites in the 3' UTR of the CTLA-4 gene and APECED-related AIRE gene mutations analysis in sporadic idiopathic hypoparathyroidism. AB - Autoimmune diseases such as Graves' disease and type 1 diabetes have been linked with +49A/G and CT60 single nucleotide polymorphisms (SNPs) in the 3' UTR of the cytotoxic T-lymphocyte antigen-4 (CTLA-4) gene. Both these SNPs are functionally relevant and linked with T-lymphocyte activation. Hypoparathyroidism is seen in 70% of patients with autoimmune polyendocrinopathy candidiasis ectodermal dystrophy syndrome (APECED). Although calcium sensing receptor autoantibodies (CaSRAb) and generalized activation of T lymphocytes are reported among patients with sporadic idiopathic hypoparathyroidism (SIH), CTLA-4 gene SNPs and APECED related autoimmune regulator (AIRE) gene mutations have not been assessed in them. We studied lead CTLA-4 gene SNPs and APECED-related AIRE gene mutations in 73 patients with SIH and 114 healthy subjects. The CTLA-4 gene SNPs +49A/G in exon 1, CT60A/G in 3' UTR and -318C/T in the promoter region were genotyped by polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) using BstEII, NcoI and MseI endonucleases, respectively. The APECED-related AIRE gene mutations, which is R257X (Finn-major) in exon 6, 4-bp insertion and 13-bp deletion in exon 8, and Iranian Jews population 'Y85C' mutation in exon 2, were studied by PCR-RFLP (Taq-I), PCR and nucleotide sequencing, respectively. CaSRAb were studied by immunoblotting. The frequencies of CTLA-4 A/A(49), A/G(49) and G/G(49) genotypes in the patients (47.9%, 38.4% and 13.7%) and controls (45.6%, 39.5% and 14.9%, respectively) and the frequencies of CT60 A/A, A/G, and G/G genotypes in the patient (42.4%, 37.0% and 20.6%) and the control (38.6%, 40.4% and 21.0%, respectively) groups were not significantly different. The frequencies of various haplotypes including genetic loci +49A/G and CT60 and frequencies of G alleles at these positions were comparable between patient and the control groups and its presence did not correlate with clinical and biochemical indices of the disease. None of the patients had APECED-related AIRE gene mutations. Lack of significant difference in the pattern of CTLA-4 A/G(49) and/or CT60A/G genotypes and absence of common APECED syndrome-related AIRE gene mutations among patients and controls suggest that these sites do not play a role in the development of the SIH. PMID- 16313306 TI - A NFKB1 promoter polymorphism is involved in susceptibility to ulcerative colitis. AB - Nuclear factor kappaB (NF-kappaB) designates a group of critical transcription factors involved in a variety of immunologic and/or inflammatory processes. Conceivably, genes involved in the NF-kappaB pathway make interesting candidate genes for chronic inflammatory disorders, including the inflammatory bowel diseases (IBD), Crohn's disease (CD) and ulcerative colitis (UC). In two mouse models of colitis, strong linkage has been observed with a locus on chromosome 3 that harbours the Nfkb1 gene. In addition, a polymorphism in the promoter region of the human NFKB1 gene was found to be associated with susceptibility to UC. In this study, we searched to confirm this previously found association in IBD in a different population. Allele and genotype frequencies of the -94 ins/delATTG polymorphism were determined in 266 unrelated Dutch Caucasian IBD patients (127 UC, 139 CD), and 155 matched healthy controls. The allele frequency of the deletion was significantly higher in UC patients (P = 0.019), but not in CD patients, compared to healthy controls, and the UC patients homozygous for the 94 ATTG deletion had a younger age of onset. Our findings confirm the previously found association between this polymorphism and susceptibility to UC in an independent study population and adds further evidence for the role of this gene in disease susceptibility. PMID- 16313307 TI - Haplotyping of the canine MHC without the need for DLA typing. AB - The genomic matching technique has proven useful in MHC haplotyping in humans. We have adopted a similar approach in Australian cattle dogs and report that genotyping can be achieved with a single assay. PMID- 16313308 TI - Variability in TRBV haplotype frequency and composition in Caucasian, African American, Western African and Chinese populations. AB - The polymorphic T-cell receptor Vbeta (TRBV) genes encode much of the variable region of the T-cell receptor beta chain. Analysis of allele frequencies of three closely linked polymorphic TRBV genes, TRBV7-3, TRBV9 and TRBV6-4, was undertaken in several populations. The frequencies of these alleles are not significantly different in populations of Caucasians, African Americans and Western Africans. However, Chinese population is extremely homogenous at all three loci. The current study identifies the existence of haplotypic relationships between alleles of these genes in the Caucasian population. The ORF allele TRBV7-3*A3 is found exclusively on chromosomes bearing TRBV9*A2 and TRBV6-4*A2 in this cohort. In contrast, TRBV7-3*A1 and the null allele TRBV7-3*A2 are associated only with TRBV9*A1 and TRBV6-4*A1. This pattern of linkage disequilibrium (LD) is altered in the African American and Western African populations. In these cohorts, there is a marked reduction in LD between alleles of TRBV7-3 and TRBV9. This study is consistent with previous population genetic studies wherein African-derived samples have a greater level of genetic diversity compared to Caucasians. These data also demonstrate that patterns of LD are not consistent across the entire TRBV locus. PMID- 16313309 TI - Polymorphism of the DPB1 locus in Hani ethnic group of south-western China. AB - Polymorphism of HLA-DPB1 was revealed with a sequencing-based typing (SBT) method in 47 unrelated healthy individuals from Yunnan Hani ethnic minority. The alleles DPB1*5901 and DPB1*7001 were detected for the first time in Chinese populations. A dendrogram showed that the Hani ethnic group belongs to the southern group of Chinese. PMID- 16313310 TI - Nomenclature for factors of the HLA system, updated July 2005. PMID- 16313311 TI - Nomenclature for factors of the HLA system, update August 2005. PMID- 16313312 TI - Nomenclature for factors of the HLA system, update September 2005. PMID- 16313313 TI - Use of IL-2 receptor antagonists to reduce delayed graft function following renal transplantation: a review. AB - Delayed graft function (DGF) occurs in approximately 30% of renal transplant patients, and significantly increases risk of long-term graft loss. This article reviews the potential for use of interleukin-2 receptor (IL-2R) antagonists to reduce the burden of DGF. IL-2R antagonists decrease incidence of acute rejection without increasing risk of cytomegalovirus infection or malignancy, and show equivalent efficacy to lymphocyte-depleting antibody agents in standard risk patients with immediate graft function. The nephrotoxicity associated with calcineurin inhibitors (CNIs) has led to use of delayed or low-dose CNI regimens with induction therapy in patients with DGF. In this setting, use of an IL-2R antagonist with mycophenolate mofetil and steroids with delayed cyclosporine appears to be associated with a low incidence of biopsy-proven rejection and comparable renal function to patients with immediate function. Additionally, there is intriguing evidence to suggests that IL-2R antagonists may reduce risk of DGF occurring. A number of large-scale and smaller studies have reported a trend to reduced incidence of DGF or improved early renal function using IL-2R antagonists compared with placebo, although data are not entirely consistent. In conclusion, the ability of IL-2R antagonists to reduce acute rejection with no additional safety concerns makes them an attractive option for patients with DGF. PMID- 16313315 TI - Successful combined liver and kidney transplant for COACH syndrome and 5-yr follow-up. AB - The COACH syndrome is a very rare disorder with cerebellar vermis hypoplasia, oligophrenia, ataxia, coloboma, and hepatic fibrosis. Nineteen cases with COACH diagnosis have been reported. Neurologic abnormalities are the first symptoms in most cases. Complications of the hepatopathy [portal hypertension, esophageal varices, and gastrointestinal (GI) bleeding] contribute extensively to the morbidity and lethality in the course of the disease. We describe a 28-yr-old female with COACH syndrome resulting in chronic renal and hepatic insufficiency. The patient was found to have significant mental retardation, truncal ataxia, motor abnormality and occulomotor abnormality. She began to develop GI bleeding and encephalopathy because of biopsy-confirmed cirrhosis. We performed combined liver and kidney transplant after challenging discussion. Her postoperative course was uneventful, and she was discharged on the ninth postoperative day (POD). She has not had any problems at 1, 3 and 5-yr follow-up with excellent liver and renal function. This is the first description of successful combined liver and kidney transplant with long-term follow-up. The decision for transplant is challenging because COACH syndrome is rare with only descriptive characterization and patients have non-progressive ataxia and mental retardation. However, our case shows that liver and kidney transplant can be medically successful, and the individuals achieve long-term success if they have a stable neurological condition and an excellent support system. PMID- 16313314 TI - Consequences of vancomycin-resistant Enterococcus in liver transplant recipients: a matched control study. AB - BACKGROUND: Liver transplant recipients are at high risk for multi-drug resistant infections because of broad-spectrum antibiotic and immunosuppression. This study evaluates the clinical and financial impact of vancomycin resistant Enterococcus (VRE) in liver transplant recipients. METHODS: Liver transplant recipients with VRE from 1995 to 2002 were identified and matched (age, gender, UNOS status, liver disease and transplant date) to controls. Demographics, clinical factors, co-infections, antibiotic use, length of stay, abdominal surgeries, biliary complications, survival and resource utilization were compared with matched controls. RESULTS: Nineteen patients were found to have 28 VRE infections via evaluation of microbiologic culture results of all liver transplant patients in the transplant registry. Thirty-eight non-VRE patients served as matched controls. The four most common sites VRE was cultured from included blood (35%), peritoneal fluid (35%), bile (20%), and urine (12%). Median time from transplant to infection was 48 d (range of 4-348). No significant differences in demographics were observed. The VRE group had a higher incidence of prior antibiotic use than the non-VRE group (95% vs. 34%; p < 0.05). The VRE group also experienced more abdominal surgery (20/19 vs. 3/38; p = 0.029), biliary complications (9/19 vs. 9/38; p = 0.018) and a longer length of stay (42.5 vs. 21.7 d; p = .005). Survival in the VRE group was lower (52% vs. 82%; p = 0.048). Six of the 19 VRE patients were treated with linezolid for eight infection episodes, and four of six patients survived. Eight patients were treated with quinupristin/dalfopristin for nine infections, and two of eight survived. Increased cost of care was observed in the VRE group. Laboratory costs were higher in the VRE group (6500 dollars vs. 1750; p = 0.02) as well. CONCLUSION: VRE was associated with prior antibiotic use, multiple abdominal surgeries, biliary complications and resulted in decreased survival compared to non-VRE control patients. VRE patients also utilized more hospital resources. Linezolid showed a trend toward improved survival. PMID- 16313316 TI - Living and deceased donor liver transplantation for unresectable hepatoblastoma at a single center. AB - Hepatoblastoma (HB) is the most common malignant liver tumor in children. The application of living donor liver transplantation (LDLT) in the management of unresectable HB may add new therapeutic opportunities. We evaluated the outcomes of patients who underwent liver transplantation for treatment of unresectable HB in the period between August 1985 and June 2003. Ten children had a diagnosis of unresectable HB. Mean age at transplantation was 5.8 yr. Eight patients were transplanted with deceased donor grafts. Two patients underwent LDLT. Pre transplant chemotherapy was used in 90% of cases. Post-transplant survival ranges from 3.7 to 18.6 yr. Three patients died of recurrent disease at 4, 14 and 38 months. The two LDLT recipients were able to get pre-transplant chemotherapy with a rapid decision towards transplantation; both are alive and well at 5.5 and 11 yr post-transplant. Our experience supports the role of LDLT and deceased donor liver transplantation in the management of unresectable HB when waiting times can be detrimental to the patient's survival. PMID- 16313317 TI - Systemic strategies for chemoprevention of skin cancers in transplant recipients. AB - BACKGROUND: Solid organ transplant recipients (OTRs) are a growing population at high risk for cutaneous neoplasms, resulting in significant post-transplant morbidity and mortality. Management of malignant and pre-malignant cutaneous lesions in transplant recipients is challenging, making prevention of such neoplasms paramount. The objectives of the present study are to review and analyze systemic strategies for chemoprevention of malignant and pre-malignant cutaneous neoplasms in OTRs. METHODS: MEDLINE and PubMed searches were performed to identify studies with original data quantifying the effects of systemic agents on the development of malignant cutaneous neoplasms in patients with solid organ transplants. RESULTS: We identified nine studies describing 111 transplant recipients that quantified the effects of oral retinoids on cutaneous neoplasms. A majority of the studies found a decrease in the number of malignant and pre malignant cutaneous lesions in patients treated with systemic retinoids, with several studies noting increased benefit in those patients with multiple previous skin cancers. Multiple studies described a rebound effect, with increased numbers of neoplasms occurring following discontinuation of retinoids. Side effects often limited dosing, but required discontinuation of retinoids in a minority of patients. No studies were identified that adequately quantified the effects of other systemic agents on skin cancer incidence in this population. CONCLUSIONS: Although systemic retinoids are frequently used for chemoprevention of cutaneous malignancies in OTRs, the data supporting their use are composed largely of small uncontrolled case reports and case series. However, the available data suggest that retinoids have chemopreventative effects in this population. Although optimal dosing and indications for initiation of systemic retinoid therapy are not conclusive from the data, it suggests that retinoids are most effective in patients with multiple previous non-melanoma skin cancers. Side effects and beneficial effects were noted across a wide range of doses, suggesting that retinoids should be initiated at a low dose and increased as tolerated to a minimally effective dose. Further investigation through randomized controlled trials is needed to further clarify the tolerability and efficacy of multiple dosing regimens on the incidence of pre-malignant and malignant lesions in transplant recipients. The therapeutic role of other systemic agents in the transplant population has not been established. PMID- 16313318 TI - Skin cancer in renal allograft recipients--experience in different ethnic groups residing in the same geographical region. AB - Cancer is a recognized long-term complication of kidney transplantation. Skin cancer is the most common post-transplant malignancy in developed countries but information is limited on the nature of skin cancer in allograft recipients from developing countries followed up over an extended period. The records of all patients (n = 542) who had received kidney transplants (n = 623) at our institution over a 23-yr period were reviewed and those with skin cancer were identified. Demographic, clinical, and pathologic details were collected. After a mean follow up of 6.3 yr 11 (5.9%) white patients had skin cancer of whom nine (82%) were male. No non-whites had skin cancer. In white patients skin cancer accounted for 68% of all post-transplant malignancies. Squamous cell carcinoma was the most common malignant skin lesion and 84% of all lesions occurred in sun exposed areas. Specific immunosuppression did not appear to influence the number of lesions or the interval from transplantation to cancer development. Patients responded well to treatment with no mortality related to the skin cancer. Skin cancer is relatively unique to patients of European origin. PMID- 16313319 TI - Chronologically different impacts of immunologic and non-immunologic risk factors on renal allograft function. AB - INTRODUCTION: Upon analysis of the risk factors affecting renal graft survival and function, the time-dependent effects of each risk factor should be differentiated from their net effects. To evaluate the chronologically different impacts of risk factors on graft renal function, we reviewed 390 recipients who received a kidney from 1-haplotype-matched living-related donors. MATERIALS AND METHODS: Until 5-yr post-transplantation (TX), yearly serum creatinine (Scr), 24 h urinary excretion of protein, and their yearly changes were compared by the episodes of acute rejection within 1 yr, the kidney weight to recipient body weight (KW/BW) ratio, the donor/recipient (D/R) age ratio, and the D/R gender pairing. The Kaplan-Meier method, Cox proportional hazard model, ANOVA, and repeated measures ANOVA were each applied for different purposes. RESULTS: Only the episodes of acute rejection were a significant risk factor affecting graft survival. The episodes of acute rejection, KW/BW ratio, D/R age ratio, and D/R gender pairing consistently and independently had significant influences on Scr. Recipients having the lowest KW/BW ratio (first quartile) or the highest D/R age ratio (fourth quartile) had rapid increments of Scr after 4-yr post-TX. After 3 yr post-TX, there were significant correlations between the number of non immunologic risk factors present and the yearly changes in Scr. CONCLUSIONS: Non immunologic factors had a detrimental effect on renal graft function, especially after 3-yr post-TX. If immunologic risks seem to be similar, size matching, age, and gender pairing should be considered for better long-term graft function in renal TX recipients. PMID- 16313320 TI - Incidence and clinical significance of human parvovirus B19 infection in kidney transplant recipients. AB - Human parvovirus B19 (B19) infection has been known to cause chronic anemia, pure red cell aplasia (PRCA), glomerulopathy, and allograft dysfunction in kidney transplant (KT) recipients. The aim of this study was to evaluate the incidence and clinical significance of B19 infection in KT recipients. A total of 537 serum samples from 167 KT recipients were included in the present study. The incidence of B19 infection was based on either qualitative polymerase chain reaction (PCR) or quantitative PCR with LightCycler Parvovirus B19 Quantitation kit. Clinical significance of B19 infection was investigated by a retrospective review of hemoglobin (Hb) levels and the results of kidney and bone marrow biopsies. The overall PCR positive rate was 18.3% (98/537), and 52 of 167 (31.1%) KT recipients showed at least one positive PCR. In addition, 20 of 167 subjects (12.0%) showed PCR-positivity more than two consecutive times, and they had significantly lower Hb levels than those with negative or one positive PCR (p < 0.0001). Furthermore, two patients suffered from PRCA, which was confirmed by bone marrow biopsy. However, B19 infection did not seem to affect the graft outcome. In conclusion, the B19 infection in KT recipients was not uncommon and was associated with low Hb levels and PRCA after KT. PMID- 16313321 TI - Predictors of health-related quality of life in hypertensive recipients following renal transplantation. AB - BACKGROUND: Health-related quality of life (HRQoL) improves after renal transplantation. However, it is unclear which variables are the strongest determinants of HRQoL following renal transplantation. In this study, we wanted to assess whether antihypertensive medication, donor type, human leukocyte antigen (HLA)-compatibility or other variables could predict HRQoL 6-12 months after transplantation. METHODS: The study was a follow up of 124 patients recruited to a single center, randomized, double-blind clinical trial, comparing the effects of lisinopril and nifedipine in hypertensive renal transplant recipients. HRQoL was assessed with the Short Form 36 (SF-36) questionnaire. Bivariate and multiple linear regression analysis were used to assess the relationship between potential predictors and the physical component summary (PCS) and mental component summary (MCS) scales of the SF-36. RESULTS: Average scores 6-12 months after transplantation did not differ between patients randomized to lisinopril or nifedipine, or between cadaveric and living donor recipients on any of the eight SF-36 scales, or the two summary scales. In multivariate analyses, recipient age (p = 0.01) and cold ischemia time >14.5 h (p = 0.04) were independent predictors of the PCS score. Recipient age (p = 0.05), 2 4 HLA-AB mismatches (p = 0.05) and donor age (p = 0.03) were independent predictors of the MCS score. CONCLUSIONS: There was no evidence of differences in HRQoL according to lisinopril or nifedipine, or living vs. cadaveric donor transplantation. HRQoL was significantly reduced with longer cold ischemia time and more than one HLA-AB mismatches, after adjusting for age. These donor kidneys related issues need confirmation. PMID- 16313322 TI - Hepatitis C virus infection in renal transplant patients: a comparative study with immunocompetent patients. AB - The behavior of hepatitis C in states of immunodeficiency is poorly understood and it is still unclear whether the characteristics of hepatitis C virus (HCV) infection in renal transplant patients differ from those observed in immunocompetent subjects. The aim of this study was to compare the biochemical and histologic characteristics of chronic HCV infection between renal transplant and immunocompetent patients. Forty-one HCV-RNA-positive renal transplant patients and 41 immunocompetent controls matched for gender, age at infection and time of infection were included in the study. The groups were compared regarding laboratory and histologic variables. Renal transplant patients showed lower alanine aminotransferase (ALT) levels (p = 0.005) and higher levels of gamma glutamyltransferase (p = 0.003), alkaline phosphatase (p < 0.001), and direct bilirubin (p < 0.001) when compared with controls. Histologic analysis revealed less intense portal (p < 0.001) and periportal (p = 0.046) inflammatory infiltrate in renal transplant patients but a larger proportion of cases with confluent necrosis (p = 0.043). No difference in the presence of septal fibrosis, hepatic steatosis, bile duct injury and siderosis was observed. However, there was a difference in the presence of lymphoid aggregates, which were less frequent in the renal transplant group (p < 0.001). In conclusion, the characteristics of hepatitis C in renal transplant patients differ from that observed in immunocompetent patients. In renal transplant patients, HCV infection is biochemically characterized by lower ALT levels and higher frequency of cholestasis. Regarding histology, despite lower frequency of lymphoid aggregates and less intense portal/periportal inflammatory infiltrate, a greater lobular damage was observed. The impact of these differences on the progression of fibrosis remains to be established. PMID- 16313323 TI - Splenectomy and preemptive interferon therapy for hepatitis C patients after living-donor liver transplantation. AB - Recurrent hepatitis C after liver transplantation is a major cause of graft failure. We routinely perform preemptive interferon and ribavirin therapy in patients after living-donor liver transplantation indicated for hepatitis C related cirrhosis. One of the obstacles for the therapy includes blood cytopenia. To overcome this problem, we recently performed splenectomy concurrently with liver transplantation. Thirty-five patients underwent liver transplantation and received preemptive therapy for hepatitis C. They were divided into two groups: those with splenectomy (group A, n = 21) and those without (group B, n = 14). There was no significant difference in the frequency of morbidity between the groups. Platelet counts were well maintained in group A patients during the therapy, and cytopenia led to the discontinuation of the therapy in one group B patient. The results of the preliminary study warrant a randomized control trial to examine the feasibility of splenectomy and preemptive viral therapy during liver transplantation for hepatitis C. PMID- 16313324 TI - Incidence and prevalence of diabetes mellitus in patients with cystic fibrosis undergoing lung transplantation before and after lung transplantation. AB - Cystic fibrosis (CF) related diabetes mellitus (DM) occurs in 15% of adult pancreatic insufficient CF patients. Lung transplantation is a treatment option for end-stage CF. We hypothesized that the prevalence of DM increases after lung transplantation. The study population included adult patients undergoing lung transplantation from March 1988 to March 2002 for end-stage CF at the University of Toronto. Demographic data, exocrine pancreatic function, presence of DM before and after transplant, as well as timing of its development after transplant were collected. Eighty-six patients met the study criteria; 77 of 86 (89.5%) of patients were pancreatic insufficient and were further analyzed. Median follow-up post-transplant was 3.3 yr (interquartile range: 1.2-7.2). Their mean age was 29.7 +/- 8.1 yr and 46 of 77 (59.7%) were male. The prevalence of DM increased from 22 of 77 (28.6%) before transplant to 38 of 77 (49.4%) after transplant (p = 0.008). The median time of DM development after transplant was 80 d (range: 13 4352). Sixteen of 55 (29.1%) of pancreatic insufficient patients who were non diabetic prior to transplant, developed DM after transplant. DM is common in CF patients undergoing lung transplantation and the prevalence increases after transplant. PMID- 16313325 TI - Impact of mycophenolate mofetil (MMF)-related gastrointestinal complications and MMF dose alterations on transplant outcomes and healthcare costs in renal transplant recipients. AB - Mycophenolate mofetil (MMF), a mycophenolic acid prodrug, is a highly effective adjunct immunosuppressive agent in transplant therapy. Although MMF is generally well tolerated, optimal therapy may be limited by adverse effects, in particular gastrointestinal (GI) toxicity, which has been reported to occur in up to 45% of MMF-treated patients. MMF dose changes resulting from these adverse events may lead to sub-therapeutic dosing and impaired clinical outcomes. This retrospective study analyzed clinical records from 772 renal transplant patients from 10 US transplant centers who were initiated on MMF. The analysis revealed that 49.7% (n = 382) of patients experienced at least one GI complication within the first 6 months post-transplant, with 66.8% (n = 255) of these having multiple GI complications. Of the patients with GI complications, 39.0% experienced MMF dose adjustments or discontinuation of MMF therapy. Patients with GI complications who experienced MMF dose adjustments/discontinuation had a significantly increased incidence of acute rejections compared with patients without GI complications (30.2% vs. 19.4%; p = 0.005). Mean treatment costs were higher in patients with GI complications than in those with no GI complications, particularly in those who experienced MMF dose adjustments/discontinuation (p = 0.0001). The mean incremental cost for patients experiencing GI complications was US$3700 per patient during the 6 months post-transplant (p < 0.001), which was mainly attributable to hospitalization costs. In summary, GI complications and MMF dose adjustments/discontinuations are associated with a significant negative impact on transplant outcomes and markedly increase short-term treatment costs. PMID- 16313326 TI - C4d as a significant predictor for humoral rejection in renal allografts. AB - OBJECTIVE: To determine the diagnostic and clinical significance of C4d accumulation in renal allografts followed by acute rejection. METHODS: A total of 158 graft biopsies performed from December 1997 to December 2002 were classified, according to the Banff-97 criteria, into hyperacute rejection (HAR, three cases), acute vascular rejection (AVR, 27), acute cellular rejection (ACR, 24), borderline rejection (BR, 38), acute tubular necrosis (ATN, five), stable graft function (SGF, 30) and baseline kidney (31). Immunohistochemical technique was used to determine the C4d deposition level. RESULTS: The percentages of C4d positive in HAR, AVR, ACR, BR, ATN, SGF and baseline kidney groups were 100% (3/3), 77.8% (21/27), 37.5% (9/24), 23.7% (9/38), 0% (0/5), 3.3% (1/30), 0% (0/31), respectively. In acute rejection patients, the peak serum creatinine (sCr) level in C4d(ptc)-positive group (41 cases) was 334.82 +/- 238.37 micromol/L, with that of C4d(ptc)-negative group (47 cases) being 220.20 +/- 176.94 micromol/L (p < 0.01). After treatment, the trough sCr level in C4d(ptc) positive group and C4d(ptc)-negative group were 176.87 +/- 111.80 and 121.75 +/- 34.59 micromol/L (p < 0.01), respectively. In each AVR, ACR and BR subgroups, the peak sCr level, the trough sCr level, after 3 or 6 months of AR, the sCr level in C4d(ptc)-positive subgroup was higher than that of C4d(ptc)-negative subgroup. There were more resistance against steroid therapy [65.9% (27/41) vs. 36.2% (17/47), p = 0.005] and a higher rate of graft loss [29.3% (12/41) vs. 6.4% (3/47), p = 0.001] in C4d(ptc)-positive group than those of C4d(ptc)-negative group. In each C4d(ptc)-positive subgroup of AVR, ACR and BR the complete reversion was 57.1, 56 and 66.7%, respectively, it is almost same. CONCLUSION: The C4d deposition level is of great value in diagnosis of acute rejection caused by humoral immune components. It is a significant predictor of graft survival and will be of great help when treating acute rejection. PMID- 16313327 TI - Outcomes of pancreas transplants for patients with type 2 diabetes mellitus. AB - BACKGROUND: The objective of this study was to examine how effectively pancreas transplants provide long-term glucose control in patients with type 2 diabetes mellitus (DM). We used guidelines from the American Diabetes Association (ADA) and the World Health Organization (WHO) to appropriately classify recipients with type 2 DM (vs. type 1 DM). RESULTS: From 1994 through 2002, a total of 17 patients with type 2 DM underwent a pancreas transplant at our center. Mean recipient age was 52.5 yr. The mean age at diabetes onset was 35.7 yr; mean duration, 16.8 yr. Most recipients had one or more secondary complications related to their diabetes: retinopathy (94%), neuropathy (76%), or nephropathy (65%). At the time of their transplant, three (18%) were on oral hypoglycemic agents alone and 14 (82%) were on insulin therapy. Of the 17 transplants, seven (41%) were a simultaneous pancreas-kidney transplant (SPK); four (24%), pancreas after kidney transplant (PAK); and six (35%), pancreas transplant alone (PTA). One recipient died during the perioperative period because of aspiration. The other 16 recipients became euglycemic post-transplant and had a functional graft at 1 yr post-transplant (patient and graft survival rates, 94%). Now, with a mean follow-up of 4.3 yr post-transplant, the patient survival rate is 71%. The four additional deaths were because of sepsis (n = 2), suicide (n = 1), and unknown cause (n = 1). All four of these recipients were insulin-independent at the time of death, although one was on an oral hypoglycemic agent. Of the 12 recipients currently alive, 11 remain euglycemic without requiring insulin therapy or oral hypoglycemic agents; one began insulin therapy 1.2 yr post-transplant (current daily dose, 60 units). CONCLUSION: These findings suggest that pancreas transplants can provide excellent glucose control in recipients with type 2 DM. All 16 (94%) of our recipients whose transplant was technically successful were rendered euglycemic. Long-term results were comparable with those seen in transplant recipients with type 1 DM. PMID- 16313328 TI - Ursodeoxycholic acid treatment in isolated chronic graft-vs.-host disease of the liver. AB - OBJECTIVES: Data regarding the long-term treatment of ursodeoxycholic acid (UDCA) in individuals of chronic graft-vs.-host disease (cGVHD) of the liver are limited. The aims of this prospective study were to determine whether, (i) UDCA treatment is useful as a long-term treatment for individuals with limited cGVHD of the liver following allogeneic hematopoietic cell transplantation, and (ii) the tolerability of UDCA treatment in such individuals. METHODS: Fifteen consecutive patients with de novo isolated cGVHD of the liver were included. All individuals were treated with UDCA at a dose of 13 mg/kg/d for 1 yr. Clinical evaluation and laboratory testing were assessed at 30-d intervals during UDCA therapy and every 30 d after discontinuation of UDCA for a total of 3 months. RESULTS: At the end of the treatment, 60% of patients with cGVHD of the liver had normal liver tests, the remaining 40% of patients demonstrated improvement in their abnormal liver tests (partial responders), whereas none of the patients had worsening of the liver tests. When compared with baseline, there was a significant decrease in the serum aminotransferases, alkaline phosphatase and gamma-glutamyl transpeptidase levels after completion of the UDCA treatment at 12 months (p < 0.01). No significant increase in serum liver enzyme tests was observed at the third month after the completion of therapy. Pruritus in seven of nine patients resolved after UDCA treatment. All patients completed their assigned treatment with no major adverse event. CONCLUSION: Long-term UDCA therapy appears to be effective, safe and tolerable in individuals with cGVHD presenting with isolated liver involvement. PMID- 16313329 TI - Coagulation and fibrinolytic profiles and appropriate use of heparin after living donor liver transplantation. AB - Heparin is widely used to reduce the incidence of vascular thrombosis after liver transplantation. Appropriate use of heparin based on changes in coagulation and fibrinolytic profiles, however, has not yet been discussed in detail. We performed living-donor liver transplantation for 128 adult patients. In this series, dalteparin (25 IU/kg/d) was administered until post-operative day (POD) 2. On POD 3, the anticoagulant drug was changed to heparin (unfractionated heparin sodium, 5000 U/d), the dose of which was changed according to the level of activated clotting time (ACT) targeted between 130 and 160 s. The plasma level of plasmin-alpha2 plasmin inhibitor complex, thrombin-antithrombin III complex (TAT), and fibrin degradation product D-dimer (FDP-DD) were monitored in the 21 patients. Predictors for heparin doses were analyzed among clinical parameters (n = 128). Four patients (3%) were complicated with thrombosis despite the above mentioned anticoagulation protocol. Transfusion and/or relaparotomy for hemostasis were necessary for bleeding in 19 patients (15%). The TAT level markedly elevated until POD 3 and FDP-DD peaked later. The required heparin dose to maintain adequate ACT levels increased linearly until POD 8, and kept constant thereafter, which correlated with the weight of the liver graft (p = 0.01). Thus, frequent monitoring of the heparin dosage is necessary to keep the ACT level in the target range in the first post-operative week. High hemorrhage complications in our series indicate that the lower target ACT range may be preferable in the second post-operative week. PMID- 16313330 TI - Interstitial expression of heat-shock protein 47 correlates with capillary deposition of complement split product C4d in chronic allograft nephropathy. AB - BACKGROUND: Chronic allograft nephropathy (CAN), associated with late-allograft dysfunction is caused by alloantigen-dependent and -independent mechanisms, and eventually leads to interstitial fibrosis (ci). Activation of complement cascade is considered to be a poor prognostic marker of graft survival. This study was designed to examine the relationship between the expression of C4d and heat-shock protein 47 (HSP47, a collagen-specific chaperone) in the development of interstitial fibroproliferative lesions in CAN. METHODS: Sixty-three renal allograft biopsy specimens, obtained from 48 patients, were examined for the expression of C4d, HSP47, CD68 and alpha-smooth muscle actin (alpha-SMA) by immunohistochemistry. Double-staining was performed to determine the colocalization of C4d and HSP47. The relationship of between the expression of C4d, HSP47, CD68 and alpha-SMA and the clinical and histopathological parameters were statistically analysed. RESULTS: No expression of C4d was noted in the tubulointerstitium including peritubular capillary (PTC) of the control kidney. C4d was expressed in PTC in one-third of allograft renal tissues with morphological evidences of CAN. The interstitial cells around the fibrotic areas of the PTC of CAN were positive for the expression of HSP47. The deposition of C4d in PTC correlated with interstitial expression of HSP47 around the PTC. Most HSP47 expressing cells were phenotypically altered myofibroblasts, as determined by the dual staining of alpha-SMA. CONCLUSIONS: The increased expression of HSP47 positively correlated with the expression of C4d in PTC, and might contribute to the progression of interstitial ci in CAN. PMID- 16313331 TI - Mast cell numbers and protease expression patterns in biopsy specimens following renal transplantation from living-related donors predict long-term graft function. AB - In human kidney transplantation the main cause of declining long-term graft function is chronic allograft nephropathy (CAN). Recent studies have implicated human mast cells (MC) in chronic inflammation and fibrosis, MC can be subtyped according to protease content: MC(T) containing tryptase only and MC(TC) containing both tryptase and chymase. We investigated immunohistochemically whether numbers and subtypes of MC in biopsy specimens 100 d after transplantation could predict subsequent fibrosis and graft dysfunction. The total number of MC/high-power field at 100 d after transplantation correlated significantly with change in creatinine clearance (DeltaCcr), defined as (Ccr at 100 d) - (Ccr at 3 yr) (R = 0.597, p = 0.0021); fibrosis index (FI) at 100 d (R = 0.583, p = 0.0066); and DeltaFI, defined as (FI at 3 yr) - (FI at 100 d) (R = 0.406, p < 0.05). The ratio of MC(TC) to total MC at 100 d also correlated with DeltaCcr (R = 0.491, p = 0.0148), FI at 100 d (R = 0.527, p = 0.0081), and DeltaFI (R = 0.417, p < 0.05). Thus, increases in number of total MC and the ratio of MC(TC) to total MC in early biopsy specimens were related to decline of long-term graft function and fibrosis. PMID- 16313332 TI - Anemia in kidney transplanted patients. AB - BACKGROUND: Although a known cardiovascular risk factor, anemia in the renal transplant recipients has only recently been receiving an increasing attention. METHODS: In a cross-sectional study, data was obtained from 959 patients followed at a single outpatient transplant clinic. Based on the guideline of the American Society of Transplantation, anemia was defined as hemoglobin (Hb) < or =130 g/L in males and < or =120 g/L in females. RESULTS: About one-third (34%) of the patients were anemic. The prevalence of anemia was comparable in males and females. Serum Hb concentration was significantly correlated with the estimated glomerular filtration rate (eGFR) (abbreviated modification of diet in renal disease formula) (r = 0.266, p < 0.001), serum transferrin (r = 0.268, p < 0.001) and serum albumin (r = 0.196, p < 0.001). None of the immunosuppressive medications or the use of angiotensin converting enzyme inhibitors was associated with a higher likelihood of anemia. In multivariate analysis the eGFR, serum albumin and serum transferrin, potential markers of nutritional status and/or chronic inflammation, and also iron deficiency were independently and significantly associated with anemia. Erythropoietin was administered only to 63 (19%) anemic patients. CONCLUSIONS: Post-transplant anemia is a prevalent and under-treated condition. Based on our results we suggest that, besides other factors, protein/energy malnutrition and/or chronic inflammation may be independently associated with anemia. Further studies are needed to determine whether the presence of anemia and its treatment will have an impact on long-term outcomes of this population. PMID- 16313333 TI - Establishing pediatric immune response zones using the Cylex ImmuKnow assay. AB - For all transplant patients, the transplant physician must balance the risk of rejection caused by under-immunosuppression against the risk of drug toxicity, secondary infections and post-transplant lymphoproliferative disorder with over immunosuppression. A Food and Drug Administration (FDA)-approved in vitro assay, the Cylex ImmuKnow assay, provides a global assessment of cellular immune function to help monitor the immune status of immunosuppressed patients. This assay uses the plant lectin phytohemagglutinin to stimulate lymphocytes; an ATP assay is then used to measure the degree of activation of CD4 T cells. However, the normal values for this assay were developed with healthy adult patients. In this study, we determined the normal ranges for the ImmuKnow assay in healthy children and compared those values to levels obtained in healthy adults and in stable pediatric renal transplant patients. We found that healthy children 12 yr of age and older showed immune function levels indistinguishable from adults, while healthy children under 12 had significantly lower immune function levels than adults. For adults, the ImmuKnow assay zones (in ng/mL ATP) of strong, moderate and low immune function correspond to >525, 225 to 525, and <225. In children under 12, we found the corresponding zones to be >395, 175-395 and <175 ng/mL. The median value for normal adults is 415, whereas it is only 295 for children <12 yr of age and this value decreases to 165 in stable renal transplant patients <12 yr of age (compared with 258 for stable adult renal transplant patients). Thus, this study provides critical information necessary to utilize the ImmuKnow assay with pediatric patients. In adults, the degree of immune function as assessed by the ImmuKnow assay helps to predict patients at risk for infection or rejection. If further studies in pediatric patients document the same and is true for children, then the ImmuKnow assay will provide a useful adjunct tool to prevent over- or under-immunosuppression as newly developed drugs are utilized or drug treatment is altered because of drug side effects, toxicity, concurrent illnesses or rejection. PMID- 16313334 TI - Treatment of X-linked childhood cerebral adrenoleukodystrophy by the use of an allogeneic stem cell transplantation with reduced intensity conditioning regimen. AB - Childhood cerebral form of X-linked adrenoleukodystrophy (X-ALD) is a rapidly progressive demyelinating condition affecting the cerebral white matter, which rapidly leads to total disability and death. The only known curative treatment for this condition is allogeneic hematopoietic stem cell transplantation (HSCT). Procedure-related toxicity is assumed to be the cause of death of patients with X ALD. Three cases of ALD successfully transplanted with the use of non myeloablative fludarabine based conditioning are described. Patients showed smooth peri-bone marrow transplantation course with fast and stable engraftment. In the 3- to 5 yr follow-up period, patients showed no deterioration in their clinical and neurological condition. Levels of very long chain fatty acids were very variable and had a tendency to decrease in at least one of the three patients. In another patient, an improvement of magnetic resonance imaging changes was found. Non-myeloablative HSCT should be considered as an early treatment for X-ALD. PMID- 16313336 TI - Osteoimmunology. PMID- 16313337 TI - Skeletal development, bone remodeling, and hematopoiesis. AB - In adult mammals, the bone marrow microenvironment is defined by close interactions between cells derived from mesenchymal progenitors and cells derived from hematopoietic progenitors. The influence that one population of cells has over the other has been a matter of intense study since it was established that hematopoietic stem cells (HSCs) require support of stromal elements to engraft, self-renew, and progress towards lineage commitment. Within the stromal components, cells of the osteoblastic lineage have the ability to interact with HSCs, and it has been proposed that they could be one of the main cell types responsible for the generation and maintenance of hematopoietic niches. Possible molecular mechanisms involved in the interaction between osteoblastic and hematopoietic cells have been described. However, understanding the relative importance of each one of them, their production by defined cells, and their kinetics of appearance have been limited by the lack of in vivo models allowing the physical and/or temporal dissection of the components of the osteoblastic lineage. Here, we provide a summary of the evidence that have established the importance of osteoblasts in hematopoiesis, and we propose new experimental strategies that could help to define the nature of these interactions. PMID- 16313338 TI - Osteoclast precursors, RANKL/RANK, and immunology. AB - Rapid progress has been made in recent years in our understanding of the mechanisms regulating the formation, activation, and survival of osteoclasts, which are derived from precursor cells in the myeloid lineage. In contrast, study of the regulation of osteoclast precursors (OCPs) has been relatively slow, in part because it has been hard to accurately identify them. However, following the discovery of cell-surface markers that facilitated purification of OCPs, recent studies have demonstrated that peripheral blood OCP numbers are increased in tumor necrosis factor (TNF)-mediated arthritis, both in animals and humans, and these numbers correlate with serum TNF levels. The increase can be reversed by anti-TNF therapy. Furthermore, the precursor cells that give rise to osteoclasts can also differentiate into other cell types, including dendritic cells. Receptor activator nuclear factor-kappaB ligand (RANKL) stimulates OCPs to produce pro inflammatory cytokines and chemokines, and RANKL blockade prevents joint inflammation in a murine model of inflammatory arthritis. These findings suggest that OCPs may serve as a source for both osteoclasts and other effector cells and participate actively in the pathogenesis of diseases. Here, we review our current understanding of the regulation of OCP formation and differentiation and provide a model of a vicious cycle in which pro-inflammatory cytokines produced in inflamed joints feedback on the bone marrow to promote the generation and release of OCPs. The OCPs then home to the inflamed joints to differentiate into mature osteoclasts or to produce more inflammatory factors in the presence of RANKL. Disruption of this cycle could provide a new strategy for the development of drugs to treat inflammatory arthritis and other disorders associated with elevated OCP/myeloid progenitors. PMID- 16313339 TI - Role of RANKL in physiological and pathological bone resorption and therapeutics targeting the RANKL-RANK signaling system. AB - Osteoclasts are primary cells for physiological and pathological bone resorption, and receptor activator of nuclear factor-kappaB ligand (RANKL) is critically involved in the differentiation, activation, and survival of these cells. Recently, therapeutics for pathological bone destruction targeting RANKL pathways has attracted a great deal of attention. Herein, we review the recent advances in the research on osteoclast biology and discuss the advantages and disadvantages of anti-RANKL therapies. PMID- 16313340 TI - Role of ITAM-containing adapter proteins and their receptors in the immune system and bone. AB - The immunoreceptor tyrosine-based activation motif (ITAM) is a highly conserved region in the cytoplasmic domain of signaling chains and receptors and is a critical mediator of intracellular signals. ITAM-mediated signals depend on the Syk or zeta-associated protein of 70 kDa tyrosine kinases, and ITAM signaling is required for the differentiation and function of B and T cells in adaptive immunity. ITAM-dependent receptors also regulate the function of innate immune cells, including natural killer cells, and myeloid-derived cells such as macrophages, neutrophils, dendritic cells, and mast cells. Myeloid lineage cells also include osteoclasts (OCLs), the cells required for bone resorption, and recent studies show a critical role for the ITAM-containing adapter proteins DAP12 and the FcRgamma chain (Fcepsilon receptor I gamma chain) in OCL differentiation. Mice deficient in both the DAP12 and FcRgamma ITAM-bearing adapters are significantly osteopetrotic with a severe defect in OCL differentiation, demonstrating the requirement for ITAM signals in bone and further implicating this pathway in the development of highly specialized cell functions in hematopoietic cells. Regulation of osteoclastogenesis by ITAM dependent receptors suggests that OCLs, similar to related myeloid cells, are tightly controlled by arrays of receptors that allow them to sense and respond to their local microenvironment like other innate immune cells. PMID- 16313341 TI - Turning down the system: counter-regulatory mechanisms in bone and adaptive immunity. AB - Major advances have been made in recent years toward the identification of transcription factors that control cell-type-specific gene expression in the skeletal and adaptive immune systems. However, the identification of factors necessary and sufficient to drive production of effector cell proteins such as matrix components and cytokines represents the first step toward understanding how cells in bone and the adaptive system achieve their highly specialized functions. Here, we provide selected examples of counter-regulatory mechanisms that serve to turn down cells involved in extracellular matrix biosynthesis and adaptive immunity at the level of the transcription factors Runx2 and nuclear factor for the activation of T cells. PMID- 16313342 TI - Role of nuclear factor-kappaB in the immune system and bone. AB - Bone metabolism is regulated by hormonal or local factors in the bone microenvironment, and recent studies have revealed that bone homeostasis is also influenced by immune system. The term 'osteoimmunology' has been proposed to explain the cross-talk between bone and the immune system. A critical element in this cross-talk is the inducible transcription factor nuclear factor-kappaB (NF kappaB), which regulates gene expression during inflammatory and immune responses. However, NF-kappaB-signaling pathways are also important for bone homeostasis, in particular for osteoclast differentiation. By bridging inflammation and bone homeostasis, NF-kappaB also contributes to the onset and progression of arthritis. Several natural compounds, synthetic drugs, and gene transfer technologies that lead to inhibition of the inhibitor of NF-kappaB kinase (IKK)/NF-kappaB activation pathway can prevent arthritis in animal models. In this review, we discuss the signaling pathway that leads to NF-kappaB activation and the role of NF-kappaB on osteoclast differentiation. Furthermore, we discuss the possibility that inhibition of NF-kappaB might provide novel therapeutic approach for inhibiting bone destruction in rheumatoid arthritis. PMID- 16313343 TI - alphavbeta3 and macrophage colony-stimulating factor: partners in osteoclast biology. AB - Osteoclasts, the sole bone-resorbing cells, arise by fusion and differentiation of monocyte/macrophage precursors. Matrix degradation requires adhesion of the osteoclast to bone, an integrin alphavbeta3-mediated event that also stimulates signals which polarize the cell and secrete resorptive molecules such as hydrochloric acid and acidic proteases. Two cytokines are necessary and sufficient for osteoclastogenesis, receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF), both produced by mesenchymal cells in the bone marrow environment. M-CSF promotes survival and proliferation of osteoclast precursors. It also contributes to their differentiation and regulates the cytoskeletal changes that accompany bone resorption. Binding of M-CSF to c-Fms, its receptor, recruits adapter proteins and cytosolic kinases, thereby activating a variety of intracellular signals. We herein review how alphavbeta3 and M-CSF, alone and in concert, impact production, survival, and function of the osteoclast, thereby controlling skeletal mass. Signals from alphavbeta3 and/or c-Fms activate Syk and Vav3, originally defined by their function in lymphoid cells. Genetic depletion of either protein generates a strong bone phenotype, underscoring the promise of osteoimmunobiology. PMID- 16313344 TI - The role(s) of Src kinase and Cbl proteins in the regulation of osteoclast differentiation and function. AB - The osteoclast resorbs mineralized bone during bone development, homeostasis, and repair. The deletion of the gene encoding the nonreceptor tyrosine kinase c-Src produces an osteopetrotic skeletal phenotype that is the consequence of the inability of the mature osteoclast to efficiently resorb bone. Src-/- osteoclasts exhibit reduced motility and abnormal organization of the apical secretory domain (the ruffled border) and attachment-related cytoskeletal elements that are necessary for bone resorption. A key function of Src in osteoclasts is to promote the rapid assembly and disassembly of the podosomes, the specialized integrin based attachment structures of osteoclasts and other highly motile cells. Once recruited to the activated integrins, especially alphavbeta3), by the adhesion tyrosine kinase Pyk2, Src binds and phosphorylates Cbl and Cbl-b, homologous multisite adapter proteins with ubiquitin ligase activity. The Cbl proteins in turn recruit and activate additional signaling effectors, including phosphatidylinositol 3-kinase and dynamin, which play key roles in the development of cell polarity and the regulation of cell attachment and motility. In addition, Src and the Cbl proteins contribute to signaling cascades that are activated by several important receptors, including receptor activator of nuclear factor kappaB and the macrophage colony-stimulating factor receptor, and also downregulate the signaling from many of these receptors. PMID- 16313345 TI - Fos/AP-1 proteins in bone and the immune system. AB - The skeleton and the immune system share a variety of different cytokines and transcription factors, thereby mutually influencing each other. These interactions are not confined to the bone marrow cavity where bone cells and hematopoietic cells exist in proximity but also occur at locations that are target sites for inflammatory bone diseases. The newly established research area termed 'osteoimmunology' attempts to unravel these skeletal/immunological relationships. Studies towards a molecular understanding of inflammatory bone diseases from an immunological as well as a bone-centered perspective have been very successful and led to the identification of several signaling pathways that are causally involved in inflammatory bone loss. Induction of receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) signals by activated T cells and subsequent activation of the key transcription factors Fos/activator protein-1 (AP-1), NF-kappaB, and NF for activation of T cells c1 (NFATc1) are in the center of the signaling networks leading to osteoclast-mediated bone loss. Conversely, nature has employed the interferon system to antagonize excessive osteoclast differentiation, although this counteracting activity appears to be overruled under pathological conditions. Here, we focus on Fos/AP-1 functions in osteoimmunology, because this osteoclastogenic transcription factor plays a central role in inflammatory bone loss by regulating genes like NFATc1 as well as the interferon system. We also attempt to put potential therapeutic strategies for inflammatory bone diseases in perspective. PMID- 16313346 TI - B cells and osteoblast and osteoclast development. AB - The molecules that regulate bone cell development, particularly at the early stages of development, are only partially known. Data are accumulating that indicate a complex relationship exists between B cells and bone cell differentiation. Although the exact nature of this relationship is still evolving, it takes at least two forms. First, factors that regulate B-cell growth and development have striking effects on osteoclast and osteoblast lineage cells. Similarly, factors that regulate bone cell development influence B-cell maturation. Second, a series of transcription factors required for B-cell differentiation have been identified, and these factors function in a developmentally ordered circuit. These transcription factors have unpredicted, pronounced, and non-overlapping effects on osteoblast and/or osteoclast development. These data indicate that at least a regulatory relationship exists between B lymphopoiesis, osteoclastogenesis, and osteoblastogenesis. PMID- 16313347 TI - The role of T lymphocytes in bone metabolism. AB - Recent findings from animal models suggest that the bone loss induced by estrogen deficiency may stem in large measure from a pathological upregulation of the adaptive immune response. While the role of activated T cells in the osteoporosis driven by inflammatory conditions and infection has been well documented, only recently has the role of T cells in the bone destruction associated with estrogen deficiency begun to be appreciated. In vivo and in vitro models of postmenopausal osteoporosis demonstrate that estrogen deficiency leads to an increase in the adaptive immune function that culminates in an increased production of tumor necrosis factor alpha (TNF) by activated T cells. TNF increases osteoclast (OC) formation and bone resorption both directly and by augmenting the sensitivity of maturing OCs to the essential osteoclastogenic factor receptor activator of nuclear factor kappaB ligand. The activation and expansion of TNF-producing T cells are key steps in estrogen deficiency-driven bone loss and are regulated by multiple interacting cytokines including transforming growth factor-beta, interleukin-7, and interferon-gamma, as well as by the process of antigen presentation. Herein, we review the experimental evidence that suggests estrogen prevents bone loss by regulating T-cell function and immune cell bone interactions. PMID- 16313348 TI - Role of interleukin-7 in bone and T-cell homeostasis. AB - Initially defined as a B-cell growth factor, the pleiotropic nature of interleukin-7 (IL-7) has increasingly become appreciated. Besides its well-known roles in B- and T-cell lymphopoiesis, IL-7 is now known to regulate the homeostasis of both mature T cells and bone cells. In bone, the precise nature of how IL-7 affects osteoclasts and osteoblasts is controversial, since it has a variety of actions in different target cells. These activities are gender specific and are dependent on whether IL-7 is delivered systemically or locally. In mature T cells, IL-7 is essential for the survival of nearly all subsets. Naive T cells are also dependent on IL-7 for survival and homeostatic proliferation in response to lymphopenia. In addition, IL-7 plays a role in the survival of memory CD8+ cells, and at high concentrations, it can compensate for the absence of IL-15. The role of IL-7 on memory CD4+ cells remains controversial and has yet to be firmly established. PMID- 16313349 TI - Interplay between interferon and other cytokine systems in bone metabolism. AB - Interferons (IFNs) play crucial roles in the regulation of a wide variety of innate and adaptive immune responses. Type I interferons (IFN-alpha/beta) are central to the host defense against pathogens such as viruses, whereas type II interferon (IFN-gamma) mainly contributes to the T-cell-mediated regulation of the immune responses. Studies of bone destruction associated with rheumatoid arthritis have highlighted the importance of the interaction between the immune and skeletal systems. Recently, a new research area, termed osteoimmunology, has been spawned by a series of studies focusing on the signaling networks between IFN and other cytokines in bone metabolisms. It has been revealed that IFN-gamma interferes with the osteoclast differentiation induced by receptor activator of nuclear factor-kappaB ligand (RANKL), and this mechanism is critical for the suppression of pathological bone resorption associated with inflammation. In addition, RANKL induces the IFN-beta gene in osteoclast precursor cells, and this induction constitutes a critical aspect of the negative feedback regulation mechanisms of RANKL signaling to suppress excessive osteoclastogenesis. Furthermore, a novel function of signal transducer and activator of transcription 1 (Stat1), the essential transcription factor for both type I and type II IFN responses, was revealed in the regulation of osteoblast differentiation. Collectively, these studies unveil novel aspects of the IFN system and indicate the operation of the intricate signaling network among IFN and other cytokine systems in bone remodeling, which might offer a molecular basis for the treatment of bone diseases. PMID- 16313350 TI - Immune responses and bone loss: the estrogen connection. AB - In addition to its effects on sexual differentiation and reproduction, estrogen has important impact on the immune system and on bone. It has also been evident that the effects of estrogen on bone to a large extent are mediated via its action on immune cells. Estrogen has a dichotomous impact on the immune system by downregulation of inflammatory immune responses but simultaneous upregulation of immunoglobulin production. Consequently, immune-mediated diseases in humans and in animal models are modulated by estrogen. Estrogen deficiency after ovariectomy in mice and after menopause in women is associated with significant bone loss. In rheumatic diseases such as rheumatoid arthritis (RA), osteoporosis is frequent, and in patients with postmenopausal RA, the degree of bone loss is dramatically increased. Hormone replacement therapy (HRT) in murine and human arthritis has beneficial effects on bone loss, as expected, but it also ameliorates inflammation and inflammation-triggered joint destruction. Long-term use of HRT has been associated with increased risk of breast cancer, thrombosis, and possibly also stroke. Accordingly, there is great need for new activators of estrogen receptors (ERs) selectively reproducing only the beneficial effects of estrogen. To achieve this aim, better knowledge of the mechanisms of how activation of ER-alpha and ER-beta modulates the immune system and bone at the cellular and molecular levels is necessary. PMID- 16313351 TI - The role of the immune system in the pathophysiology of osteoporosis. AB - The role of the immune system in the development of senile osteoporosis, which arises primarily through the effects of estrogen deficiency and secondary hyperparathyroidism, is slowly being unraveled. This review focuses on our current understanding of how the components of this complex-interlinked system are regulated and how these fit with previous models of senile and postmenopausal osteoporosis. There is certainly substantial evidence that bone remodeling is a tightly regulated, finely balanced process influenced by subtle changes in proinflammatory and inhibitory cytokines as well as hormones and cellular components that act primarily but not exclusively through the receptor activator of nuclear factor-kappaB (RANK)/RANK ligand/osteoprotegerin system. In addition, an acute or chronic imbalance in the system due to infection or inflammation could contribute to systemic (or local) bone loss and increase the risk of fracture. Although significant progress has been made, there remains much to be done in unraveling this complex interaction between the immune system and bone. PMID- 16313352 TI - Rheumatic diseases: the effects of inflammation on bone. AB - Rheumatoid arthritis, juvenile idiopathic arthritis, the seronegative spondyloarthropathies including psoriatic arthritis, and systemic lupus erythematosus are all examples of rheumatic diseases in which inflammation is associated with skeletal pathology. Although some of the mechanisms of skeletal remodeling are shared among these diseases, each disease has a unique impact on articular bone or on the axial or appendicular skeleton. Studies in human disease and in animal models of arthritis have identified the osteoclast as the predominant cell type mediating bone loss in arthritis. Many of the cytokines and growth factors implicated in the inflammatory processes in rheumatic diseases have also been demonstrated to impact osteoclast differentiation and function either directly, by acting on cells of the osteoclast-lineage, or indirectly, by acting on other cell types to modulate expression of the key osteoclastogenic factor receptor activator of nuclear factor (NF) kappaB ligand (RANKL) and/or its inhibitor osteoprotegerin (OPG). Further elucidation of the mechanisms responsible for inflammation-induced bone loss will potentially lead to the identification of novel therapeutic strategies for the prevention of bone loss in these diseases. In this review, we provide an overview of the cell types, inflammatory mediators, and mechanisms that are implicated in bone loss and new bone formation in inflammatory joint diseases. PMID- 16313353 TI - The role of immune cells and inflammatory cytokines in Paget's disease and multiple myeloma. AB - The osteoclast (OCL) is the primary cell involved in the pathogenesis of Paget's disease (PD) and the destructive bone process in multiple myeloma (MM). Both of these diseases are characterized by increased numbers of OCLs actively resorbing bone, but they differ in that bone formation is greatly increased in PD and is suppressed in MM. The marrow microenvironment plays a critical role in both disease processes, through the increased expression of inflammatory cytokines that enhance osteoclastogenesis and, in the case of MM, also suppress osteoblast (OBL) activity. In addition, the OCLs in PD are intrinsically abnormal, are markedly increased in number and size, and are hyper-responsive to inflammatory cytokines and 1,25-(OH)2D3. This article discusses the role of immune cells and inflammatory cytokines and chemokines in the increased OCL activity in PD and MM bone disease, as well as the potential role of interleukin-3 in the suppression of OBL activity in MM. PMID- 16313354 TI - Microgravity: the immune response and bone. AB - Exposure to microgravity during space flight affects almost all human physiological systems. The affected systems that are of key importance to human space exploration are the musculoskeletal, neurovestibular, and cardiovascular systems. However, alterations in the immune and endocrine functions have also been described. Bone loss has been shown to be site specific, predominantly in the weight-bearing regions of the legs and lumbar spine. This phenomenon has been attributed to a reduction in bone formation resulting from a decrease in osteoblastic function and an increase in osteoclastic resorption. In order to examine the effects of microgravity on cellular function here on earth, several ground-based studies have been performed using different systems to model microgravity. Our studies have shown that modeled microgravity (MMG) inhibits the osteoblastic differentiation of human mesenchymal stem cells (hMSCs) while increasing their adipogenic differentiation. Here, we discuss the potential molecular mechanisms that could be altered in microgravity. In particular, we examine the role of RhoA kinase in maintaining the formation of actin stress fibers and the expression of nitric oxide synthase under MMG conditions. These proposed mechanisms, although only examined in hMSCs, could be part of a global response to microgravity that ultimately alters human physiology. PMID- 16313355 TI - Relationship between SPRY and B30.2 protein domains. Evolution of a component of immune defence? AB - SPRY and B30.2 are homologous domains which can be identified in 11 protein families encoded in the human genome. These include cell surface receptors of the immunoglobulin super-family (BTNs), negative regulators of the JAK/STAT pathway (SOCS-box SSB1-4) and proteins encoded by the numerous TRIM genes. Collectively, proteins containing SPRY and B30.2 domains cover a wide range of functions, including regulation of cytokine signalling (SOCS), RNA metabolism (DDX1, hnRNPs), intracellular calcium release (RyR receptors), immunity to retroviruses (TRIM5alpha) as well as regulatory and developmental processes (HERC1, Ash2L). In order to clarify the evolutionary relationship between the two domains, we compiled a curated database of SPRY and B30.2-domain sequences. We show that while SPRY domains are evolutionarily ancient, B30.2 domains, found in BTN and TRIM proteins, are a more recent evolutionary adaptation, comprising the combination of SPRY with an additional domain, PRY. The combination of SPRY and PRY to produce B30.2 domains may have been selected and maintained as a component of immune defence. PMID- 16313356 TI - Leukotrienes modulate cytokine release from dendritic cells. AB - Leukotriene B(4) (LTB(4)) and cysteinyl leukotrienes (CysLTs) are known as potent mediators of inflammation, whereas their role in the regulation of adaptive immunity remains poorly characterized. Dendritic cells (DCs) are specialized antigen-presenting cells, uniquely capable to initiate primary immune responses. We have found that zymosan, but not lipopolysaccharide (LPS) stimulates murine bone marrow-derived dendritic cells (BM-DCs) to produce large amounts of CysLTs and LTB(4) from endogenous substrates. A selective inhibitor of leukotriene synthesis MK886 as well as an antagonist of the high affinity LTB(4) receptor (BLT(1)) U-75302 slightly inhibited zymosan-, but not LPS-stimulated interleukin (IL)-10 release from BM-DCs. In contrast, U-75302 increased zymosan-stimulated release of IL-12 p40 by approximately 23%. Pre-treatment with transforming growth factor-beta1 enhanced both stimulated leukotriene synthesis and the inhibitory effect of U-75302 and MK886 on IL-10 release from DCs. Consistent with the effects of leukotriene antagonists, exogenous LTB(4) enhanced LPS-stimulated IL 10 release by approximately 39% and inhibited IL-12 p40 release by approximately 22%. Both effects were mediated by the BLT(1) receptor. Ligands of the high affinity CysLTs receptor (CysLT(1)), MK-571 and LTD(4) had little or no effect on cytokine release. Agonists of the nuclear LTB(4) receptor peroxisome proliferator activated receptor-alpha, 8(S)-hydroxyeicosatetraenoic acid and 5,8,11,14 eicosatetraynoic acid, inhibited release of both IL-12 p40 and IL-10. Our results indicate that both autocrine and paracrine leukotrienes may modulate cytokine release from DCs, in a manner that is consistent with previously reported T helper 2-polarizing effects of leukotrienes. PMID- 16313357 TI - CD27+ B cells in human lymphatic organs: re-evaluating the splenic marginal zone. AB - The marginal zone of human spleens is regarded as an organ-specific region harbouring sessile memory B cells. This opinion has arisen by extrapolating from results obtained in mice and rats. Detection of CD27(+) B cells in situ now revealed similarities among the most superficial region of B-cell follicles in human spleens, reactive lymph nodes, inflamed appendices, tonsils and terminal ilea. The follicular surface in these organs consists of small naive immunoglobulin D (IgD)(+) CD27(-) B cells predominating in an inner area and larger IgD(+/-) CD27(+) B cells prevailing in a more superficial position. CD27(+) B cells may, however, also occupy the entire follicular periphery around the germinal centre. Together with additional peculiarities this distribution indicates a fundamental microanatomical difference among the human and rodent splenic white pulp. We hypothesize that the follicular periphery represents a recirculation compartment both for naive and memory/natural reactive B cells in all human secondary lymphatic organs. This assumption implies a difference in recirculation behaviour among human and rodent B memory cells. PMID- 16313358 TI - Evaluation of CD62L expression as a marker for vaccine-elicited memory cytotoxic T lymphocytes. AB - The development of successful vaccination strategies for eliciting cytotoxic T lymphocytes (CTLs) will be facilitated by the definition of strategies for subdividing CTLs into functionally distinct subpopulations. We assessed whether surface expression of a number of cell-surface proteins could be used to define functionally distinct subpopulations of memory CTLs in mice immunized with a recombinant vaccinia virus expressing human immunodeficiency virus (HIV)-1 envelope (Env). We found changes in cell-surface expression of CD11a, CD44, CD45RB, CD49d, CD54 and CD62L on Env-specific CD8(+) T cells that appeared to differentiate them from other CD8(+) T cells within 1 week to 1 month following immunization. Further, we saw an up-regulation of CD62L surface expression on Env specific CD8(+) memory T cells several months after immunization. However, CD62L expression did not correlate with differences in the abilities of CTLs to proliferate or produce interferon gamma (IFN-gamma) and tumour necrosis factor alpha (TNF-alpha) in vitro in response to Env peptide stimulation. Moreover, the expression of CD62L did not allow differentiation of CTLs into subpopulations with distinct expansion kinetics in vivo after adoptive transfer into naive mice and subsequent boosting of these mice with a recombinant adenovirus expressing HIV-1 Env. Therefore, the definition of memory CD8(+) T-cell subpopulations on the basis of CD62L expression in mice does not allow the delineation of functionally distinct CTL subpopulations. PMID- 16313359 TI - Differential responses of human B-lymphocyte subpopulations to graded levels of CD40-CD154 interaction. AB - Naive and memory B-lymphocyte populations are activated by CD154 interaction through cell-surface CD40. This interaction plays an important role in the regulation of the humoral immune response, and increasing evidence indicates that fine variation in CD40 binding influences B lymphocytes, macrophages and dendritic cells in murine models. Here we have investigated whether and how variations in the intensity of the CD40-CD154 interaction could contribute to differential regulation of human B-lymphocyte populations. Proliferation and differentiation of B lymphocytes were monitored in response to graded levels of CD40 stimulation in the presence of interleukin (IL)-2, IL-4 and IL-10. Our results show that the level of CD154 binding to CD40 on B lymphocytes can directly influence the evolution of CD19(+) CD27(-) and CD19(+) CD27(+) cell populations. Furthermore, proliferation, global expansion of CD19(+) cells and emergence of CD38(++) CD138(+) cells, as well as immunoglobulin G (IgG) and IgM secretion, were affected by the level of exposure of B lymphocytes to CD154. These results suggest that the CD40-CD154 interaction is more like a rheostat than an on/off switch, and its variation of intensity may play a role in the regulation of B-lymphocyte activation following the primary and/or secondary humoral immune response. PMID- 16313360 TI - Tolerosome-induced oral tolerance is MHC dependent. AB - Oral administration of a protein antigen generates a serum factor that induces tolerance when transferred into naive recipients. This serum factor has been described in rats as consisting of exosome-like structures or tolerosomes, which express major histocompatibility complex class II molecules (MHCII) and mediate antigen-specific tolerance. In this study, we investigated the functions of serum derived tolerosomes both in vivo and in vitro. Tolerosomes were purified from the 100,000 g pellet fraction of serum from ovalbumin (OVA)-fed mice. When transferred into naive recipient mice, the tolerosomes mediated OVA-specific tolerance. We also found that tolerosomes from OVA-fed mice induced the activation of OVA-specific T cells both in vivo and in vitro. The inoculation of severe combined immunodeficiency (SCID) mice with an interferon-gamma-producing cell line normalized the expression of MHCII in the intestinal epithelial cells and restored their ability to generate tolerosomes. Syngeneic but not allogeneic transfer of tolerosomes from OVA-fed donors induced tolerance in the recipients. Our results show that tolerosomes can be isolated from mouse serum, that tolerosome-induced oral tolerance requires MHCII expression in intestinal epithelial cells, and that tolerosomes are functional only in syngeneic recipients. PMID- 16313361 TI - Induction and recall of immune memory by mucosal immunization with a non-toxic recombinant enterotoxin-based chimeric protein. AB - Previous reports have suggested that peroral delivery of antigens chemically coupled to non-toxic recombinant enterotoxin B subunits, such as the cholera toxin B subunit (CTB), induces tolerance to the antigen that may be abrogated by the toxic enzyme activity of intact enterotoxins, such as cholera toxin (CT). The aim of this study was to examine the immunogenicity of a genetically coupled protein composed of the saliva-binding region (SBR) of the Streptococcus mutans surface antigen AgI/II and the non-toxic A2 and B subunits of CT (SBR-CTA2/B) compared with that of recombinant SBR admixed with CT (SBR + CT) and SBR chemically coupled to recombinant CTB (SBR-CTB) following peroral delivery by intragastric (i.g.) immunization. The results showed that i.g. immunization with SBR-CTA2/B, like SBR + CT, induced antigen-specific serum immunoglobulin G (IgG) and salivary IgA antibodies, and sensitized splenic T cells. Comparison studies with SBR-CTB produced serum IgG but not salivary IgA titres and failed to sensitize splenic cells. Immunization with SBR-CTA2/B via the intranasal route also primed for the recall of antigen-specific memory antibody responses 6 months later. These findings show that SBR-CTA2/B is an immunogenic, not tolerogenic, chimeric protein that can induce and recall antigen-specific memory responses upon mucosal immunization. PMID- 16313362 TI - Construction and characterization of a humanized anti-human CD3 monoclonal antibody 12F6 with effective immunoregulation functions. AB - 12F6 is a murine anti-human CD3 monoclonal antibody, which competes with OKT3 for binding to human T cells and possesses more effective T-cell suppression and activation properties compared to OKT3. It thus exhibits the potential to be developed as an immunoregulation agent for manipulating T-cell functions and preventing acute allograft rejection. In an attempt to minimize the immunogenicity of murine 12F6 (m12F6) for potential clinical application, a humanized version of 12F6, denoted as hu12F6, was successfully constructed by complementary determining region (CDR) grafting and shown to maintain both T-cell activation and suppression activities similar to m12F6. Furthermore, in order to reduce the first dose reaction syndrome caused by T-cell activation following the first administration of anti-CD3 antibodies, two amino acid mutations were introduced into the Fc region of hu12F6, resulting in the Fc-mutated 12F6 humanized antibody (hu12F6mu). This Fc-mutated version displayed a similar antigen-binding affinity and specificity compared with hu12F6 and m12F6 but with much weaker FcR binding activity. hu12F6mu was shown to be much less potent in the induction of T-cell proliferation, cytokine release (tumour necrosis factor alpha, interferon-gamma and interleukin-10) and early activation marker expression on the cell surface (CD69 and CD25) than parental 12F6 and OKT3 did. In contrast, hu12F6mu was effective in modulating T-cell receptor/CD3 and inhibiting mixed lymphocyte reaction with a similarity as compared to m12F6 and OKT3. In conclusion, the resultant hu12F6mu was much less mitogenic to T cells but retained potent immunosuppression, suggesting it might be an alternative to OKT3 as an immunosuppressive drug with less immunogenicity and toxicity for clinical application. PMID- 16313363 TI - Resistance to chlamydial lung infection is dependent on major histocompatibility complex as well as non-major histocompatibility complex determinants. AB - Our previous work has shown that C3H/HeN and C57BL/6 mice have differential susceptibility to Chlamydia trachomatis mouse pneumonitis (C. muridarum) lung infection. C3H/HeN (H-2(k)) mice were found to be highly susceptible to C. muridarum infection with higher mortality and more severe morbidity compared to C57BL/6 (H-2(b)) mice. To examine the role of major histocompatibility complex (MHC) genes on host resistance to chlamydial lung infection, we compared MHC congenic mice, B6.H2k [C57BL/6 background, C3H MHC (H-2(k))] and C3H.H2b [C3H/HeN background, C57BL/6 MHC (H-2(b))] and their corresponding wild type C57BL/6 mice and C3H/HeN mice, respectively, in susceptibility to C. muridarum infection. We found that B6.H2k, C3H.H2b and C3H/HeN mice are more susceptible to chlamydial lung infection compared to the wild type C57BL/6 mice by showing more serious body weight loss, higher in vivo chlamydial growth and more severe pathologic changes. Congenic B6.H2k mice showed significantly lower levels of IL-12 and IFN gamma production compared to C57BL/6 as well as C3H/HeN and C3H.H2b mice. One the other hand, although congenic C3H.H2b mice displayed similar cytokine response to C57BL/6 mice, they were highly susceptible to C. muridarum infection. Overall, the results suggest that protection against chlamydial lung infection is both MHC and non-MHC gene dependent, and that the interaction between MHC and non-MHC elements may contribute to host resistance to chlamydial infection. PMID- 16313364 TI - Cbl-b differentially regulates activation-induced apoptosis in T helper 1 and T helper 2 cells. AB - We previously reported that ligation of CD3 induces antiapoptotic signals in T helper 2 (Th2) cells, and in contrast causes Th1 cells to undergo apoptosis. Here we show that Cbl-b is accountable for the unequal response, revealing a previously unknown cell-specific regulatory function for the molecule. Absence of Cbl-b resulted in resistance to activation-induced apoptosis in murine Th1 cells following CD3 ligation, akin to what is observed in Th2 cells containing Cbl-b. Concurrent with the apoptosis profile, CD3 ligation in the absence of Cbl-b induced raft mobilization and cytoskeletal rearrangement in Th1 cells. Despite their ability to signal from CD3, Th2 cells did not aggregate their rafts, providing an explanation for cell-specific activity of Cbl-b. PMID- 16313365 TI - Selective inhibition by rottlerin of macropinocytosis in monocyte-derived dendritic cells. AB - We present here the analysis of fluid-phase endocytosis (FPE) in human blood monocytes and monocyte-derived dendritic cells (MDDC) facilitated by our serendipitous identification of rottlerin as an efficient inhibitor of dendritic cell FPE (IC(50) of 0.4 microM). Rottlerin was found to be an excellent tool for FPE analysis: rapid-acting, irreversible and selective for FPE (as opposed to receptor-mediated endocytosis) at concentrations of 3 microM and below. The inhibitory effect was not due to toxicity or visible change in membrane ruffles, but affects on cytoskeletal reorganization were evident in MDDC treated with relevant rottlerin concentrations during adhesion. A marked increase in FPE was observed in 1 hr interleukin (IL)-4 and granulocyte macrophage-colony stimulating factor (GM-CSF)-stimulated monocytes. Moreover, rottlerin inhibited the augmented FPE of 1-day cytokine treated monocytes and their augmented ability to induce T cell proliferative responses to tetanus toxoid. We conclude that rottlerin is a useful tool for investigating FPE and its functional importance. PMID- 16313366 TI - MAdCAM-1 is needed for diabetes development mediated by the T cell clone, BDC 2.5. AB - The NOD-derived islet-reactive CD4(+) T cell clone, BDC-2.5, is able to transfer diabetes to neonatal non-obese diabetic (NOD) mice but is unable to transfer disease to either adult NOD or NOD scid recipients. Transfer of diabetes to adult recipients by BDC-2.5 is only accomplished by cotransfer of CD8(+) T cells from a diabetic donor. To understand why this CD4(+) T cell clone is able to mediate diabetes in neonatal but not the adult recipients we examined the ability of the clone to traffic in the different recipients. Our studies showed that MAdCAM-1 has a very different expression pattern in the neonatal and adult pancreas. Blockade of this addressin prevents the clone from transferring diabetes to neonatal mice, suggesting that the differential pancreatic expression of MAdCAM-1 in neonatal and adult pancreas provides an explanation of the differences in diabetes development. PMID- 16313367 TI - Cathepsin S is not crucial to TSHR processing and presentation in a murine model of Graves' disease. AB - By regulating invariant (Ii) chain processing and MHC class II peptide loading, the endosomal protease cathepsin S (Cat S) has a potential role in autoimmune susceptibility. Indeed, Cat S null mice are resistant to I-Ab-restricted experimental myasthenia gravis due to inadequate peptide presentation. To explore the role of Cat S in a Graves' disease model, I-Ad-restricted wild-type (WT) and Cat S(-/-) mice were immunized with adenovirus encoding the A subunit of thyroid stimulating hormone receptor (TSHR). TSHR adenovirus immunized mice develop Th1 T cells, TSHR antibodies, and a proportion become overtly hyperthyroid. Although TSHR presentation in vitro was initially impaired in Cat S(-/-) mice, subsequent TSHR presentation in vitro and disease development were similar in both groups but with higher antibody responses in Cat S null mice. WT and Cat S(-/-) mice recognized similar T cell epitopes from a panel of overlapping TSHR peptides. TSHR responses were found to be I-Ad-restricted and Cat S(-/-) I-Ad B cells had marked defects in Ii processing. These data imply that loading of TSHR peptides critical to TSHR antibody responses becomes Ii-independent. Contrasting findings among organ-specific murine autoimmune models imply that potential uses of Cat S inhibitors to ameliorate autoimmunity must be determined empirically. PMID- 16313368 TI - NK cell activity in tuberculosis is associated with impaired CD11a and ICAM-1 expression: a regulatory role of monocytes in NK activation. AB - Although the role of natural killer (NK) cells in mycobacterial infections is unclear, it has been postulated that they contribute to protective immunity through the production of interferon (IFN)-gamma. In this study, we evaluate the effect of interleukin (IL)-10, IL-15 and IL-18 on NK lytic activity through the expression of CD16, CD11a and CD69 molecules and the induction of IFN-gamma production in patients with tuberculosis (TB) and healthy individuals (N). Our results showed an impairment of NK lytic activity and a gradual down-regulation of costimulatory and adhesion molecules on NK cells which were dependent on the severity of the disease. NK lytic activity was increased by exogenous IL-15 and IL-18 in both TB and N, and by neutralization of endogenous IL-10 only in TB; IL 15 and IL-18 increased CD69 receptor expression, while anti-IL-10 up-regulated CD16 and CD11a expression in TB. Mycobacterium tuberculosis reduced the number of intracellular adhesion molecule (ICAM)-1(+) CD14(+) cells, but in the presence of IL-15, IL-18 and anti-IL-10 its expression was up-regulated. In cells from TB patients, the observed effects of IL-15 and IL-18 on NK function were not dependent on IL-10 modulation of the surface expression of activator/adhesion molecules. In the absence of monocytes, IL-10 activated NK cells, suggesting an indirect effect on their function. Furthermore, in TB patients the depletion of monocytes increased the production of IFN-gamma by NK cells. Therefore, monocytes from TB patients regulated the NK function involving IL-10 which, through an indirect mechanism, led to the down-regulation of costimulatory/adhesion molecules and/or IFN-gamma production. PMID- 16313369 TI - Valuing professional self-regulation. PMID- 16313370 TI - Exploration of the decision-making process for inpatient hospice admissions. AB - AIM: This paper discusses some of the findings of a qualitative study which described the decision-making processes that occurred during multidisciplinary meetings when prioritizing hospice inpatient admissions. BACKGROUND: Healthcare rationing and resource allocation have been identified as important but under represented issues in the British nursing literature generally, and specifically within the field of palliative care. Little evidence currently exists about the rationing decisions made at a clinical level in hospices or palliative care units. METHOD: Adopting an ethnographic approach, data were collected at three hospice sites from three meetings at each site by observation, tape recording and transcribing of the meeting dialogue and examination of the available documentation from admissions meetings. The data were collected in 2002. RESULTS: Factors identified as potentially influential in these decisions included patient diagnosis, symptoms, current location and the stated reason for admission. The person who had assessed the patient's need for admission and whether or not a patient was personally known to a clinician present at the meeting also appeared important. The process seemed complex and incorporated different decision-making methods. Analysis of the group interactions suggested that these meetings were predominantly medically-led. CONCLUSION: Palliative care nurses need to examine their beliefs and practices in relation to how hospice inpatient care is prioritized, and develop sound evidence-based arguments in order to strengthen their role and influence in these important decisions for the benefit of patients. Further research is needed to achieve a greater understanding of these types of decision-making processes. PMID- 16313371 TI - Prostatectomy patients' postoperative pain assessment in the recovery room. AB - AIM: This paper reports a study to assess the usability and use of different pain assessment tools and to compare patients' and nurses' pain assessments in the recovery room after prostatectomy. BACKGROUND: Pain assessment is the first step towards providing adequate pain relief but poses problems because of the subjective nature of the pain experience and the lack of quantifiable measurements. Pain tools have been tested in several clinical settings, but not in the recovery room. METHODS: Data were collected in the recovery room from 45 consecutive patients who had undergone prostatectomy by asking them to evaluate their pain intensity using visual analogue scale, numeric rating scale and verbal expressions. One of two research nurses measured patients' pain at regular intervals and at the same time as the patients. Physiological parameters were also evaluated. Data were analysed as frequencies and percentages. Sum variables were formed and results were analysed using Spearman's rank correlation, Pearson's correlation and with multiple regression analysis. RESULTS: Patients varied in their ability to assess the intensity of their pain using different tools, but assessments were correlated with each other and with nurses' estimations. Nurses and patients obtained similar assessments, but nurses both underestimated and overestimated patients' pain. Patients' verbal assessments varied widely. Patients' and nurses' pain assessments showed no association with patients' pulse or mean arterial blood pressure. CONCLUSIONS: According to our results, it is not totally clear whether pain tools are usable in the recovery room. This issue calls for further research. PMID- 16313372 TI - Determining older people's needs for care by Registered Nurses: the Nursing Needs Assessment Tool. AB - AIMS: This paper reports a study to determine an appropriate framework for assessing older people's needs for nursing within the policy of 'free nursing', and to attempt to identify valid, reliable and usable assessment tool for determining this eligibility. BACKGROUND: In 2001, the United Kingdom government introduced 'free nursing care' legislation, eligibility for which was subject to a nursing needs assessment. This paper outlines the search for and subsequent development of an appropriate tool for assessing older people's eligibility for free nursing care in Northern Ireland. METHODS: Following a systematic search of the literature, existing tools used in ascertaining older people's nursing needs were identified. Each tool was systematically assessed in accordance with criteria of validity, reliability, usability, comprehensiveness of assessment and ability to quantify nursing care needs. From the findings, an assessment instrument, Nursing Needs Assessment Tool was subsequently developed and tested. Paired assessments of older people were undertaken independently by assessor dyads and evaluated statistically. Assessors' opinions on the usability of the instrument were sought through a focus group. RESULTS: One hundred and ten paired assessments were returned (63%). Overall there was 65% agreement between assessors. Kappa scores indicated good levels of inter-rater reliability. Correlation co-efficient measures reinforced these results. Findings from the focus group confirmed the validity, usability and comprehensiveness of the tool. CONCLUSIONS: The Nursing Needs Assessment Tool is a reliable, valid and usable instrument. This has major implications for the standardization of assessment for older people. PMID- 16313373 TI - Dimensions of quality in long-term care facilities in Taiwan. AB - AIM: This article presents the results of a qualitative study designed to explore the perceptions of Taiwanese elders about the quality of their care while in residence at a long-term care facility. BACKGROUND: As the population of elders in Taiwan is increasing rapidly, quality long-term care has become both a necessity and social responsibility. Research related to quality of care has focused on structure, process and outcomes as well as perceptions of quality of care and life satisfaction. Few authors have considered residents' perspectives within a cultural context. METHOD: A convenience sample of 22 residents, aged between 61 and 86 years and living in four Taiwanese care facilities, participated in semi-structured interviews. The data were generated during 2001/2002 and were analysed using content analysis. RESULTS: Six key dimensions of quality care were elicited. The dimensions were: a caring attitude, respect for individual differences, emotional support, social interaction, a supportive environment, and accessible care. These dimensions are discussed within a cultural context supportive of family connectedness and filial piety, values underpinning care expectations of elders. CONCLUSIONS: The findings provide a foundation for increased understanding of the perceived gaps between residents' aspirations about quality care and priorities identified in other studies. Additional studies based on these data are planned in order to generate a culturally relevant, psychometrically sound resident assessment tool to evaluate the quality of care from a resident perspective in long-term care facilities in Taiwan. PMID- 16313374 TI - Quality long-term care for older people: a commentary. PMID- 16313375 TI - Dynamics of hope in adults living with HIV/AIDS: a substantive theory. AB - AIM: The aim of this paper was to discuss a substantive theory on the dynamics of hope in adults living with the human immunodeficiency virus/acquired immune deficiency syndrome. BACKGROUND: Existing literature describes how the dynamics of hope have profound connections with human essence and life, wellbeing and health, suffering and illness, and nursing care, in general as well as in the context of human immunodeficiency virus/acquired immune deficiency syndrome. METHOD: The data consisted of five original articles describing the dynamics of hope in people living with human immunodeficiency virus/acquired immune deficiency syndrome and their significant others in Finland. Meta-synthesis was adapted to synthesize the original studies into a substantive theory. FINDINGS: The core category that emerged was the dynamically alternating balance between interconnected hope, despair and hopelessness based on folding (closing down of) and unfolding (opening up of) possibilities with regard to the dynamics of hope in dealing with the changing self and life with human immunodeficiency virus/acquired immune deficiency syndrome. The dynamics of hope include the subprocesses of hope, despair and hopelessness. CONCLUSION: The dynamics of hope in adults living with human immunodeficiency virus/acquired immune deficiency syndrome is an important phenomenon in nursing practice and nursing science. In taking care of people living with acquired immune deficiency syndrome, factors contributing to folding possibilities should be minimized, if possible, and factors contributing to unfolding possibilities should be nourished. The present study adds to the knowledge-base of nursing science about the dynamics of hope in people living with acquired immune deficiency syndrome. Future research should focus on testing this theory in people living with acquired immune deficiency syndrome. PMID- 16313376 TI - Small-group hypertension health education programme: a process and outcome evaluation. AB - AIM: This paper reports a study to explore the expectations, attainments and opinions of the participants, the objectives and experiences of nurse facilitators, and the changes in nurse-client relationship after a small-group hypertension health education programme. BACKGROUND: Effective control of blood pressure among hypertensive patients is essential in the reduction of cerebrovascular-related morbidity and mortality. A 'small-group hypertension education programme' has been implemented in all general outpatient clinics in Hong Kong since 2000. In order to identify what aspects of the programme were beneficial and how it benefited clients, a process evaluation study was conducted. METHODS: Following implementation of a small-group hypertension health education programme, two focus groups were run to collect data from eight nurse facilitators, and individual semi-structured interviews were conducted with 16 programme participants. The data were collected in 2001. FINDINGS: The findings indicate that both clients and nurses gained a great deal from and enjoyed the process of this health education programme, and their relationships were improved after the programme. About half of the clients reported having adopted health enhancing behaviour. The study also revealed discrepancies between the nurses and clients in their perceptions of the objectives of the programme. This indicates that there is a need to clarify the objectives with clients to minimize disappointments from unrealistic expectations. CONCLUSION: There is a need for the nurse facilitators to clarify the goal of the programme as an education group rather than as a support group. Also, needs assessment is suggested before group member recruitment in order to make the group more homogeneous. PMID- 16313377 TI - Expanding the role of the stroke nurse: a pragmatic clinical trial. AB - AIMS: This paper reports a study evaluating whether expanding a specialist nursing role to provide outreach education and support to stroke patients and carers after discharge from hospital is effective in promoting recovery. BACKGROUND: Building therapeutic relationships with patients and carers is a key component of the nursing role in stroke rehabilitation, although this is limited by the constraints of service organization. METHODS: A pragmatic randomized controlled trial was undertaken. Patients with a diagnosis of stroke were randomized to receive continued support from a stroke nurse (n = 87) or usual care and follow-up (n = 89) after discharge from hospital. Patients were recruited from two hospitals in the north-west of England from November 1999 to April 2001. Patient dependence (Barthel Index), general health (Nottingham Health Profile), activities of living (Frenchay Activity of Living Index), depression (Beck Depression Inventory) and carer strain (Carer Strain Index) were assessed at 3 and 12 months after stroke. RESULTS: The continued intervention of a stroke nurse after discharge was associated with improved patient perceptions of general health at 12 months (median difference 42.6, P = 0.012), and in particular reduced negative emotional reaction (P = 0.037) and perceived social isolation (P = 0.002). In addition, the intervention reduced carer strain at 3 months (P = 0.045), and reduced deterioration in physical dependence from 3 to 12 months (P = 0.049). CONCLUSION: The provision of continued intervention from a stroke nurse after discharge from hospital, focusing on education and support, has tangible benefits for patients and carers. PMID- 16313378 TI - Complex adaptive systems: concept analysis. AB - AIM: The aim of this paper is to explicate the concept of complex adaptive systems through an analysis that provides a description, antecedents, consequences, and a model case from the nursing and health care literature. BACKGROUND: Life is more than atoms and molecules--it is patterns of organization. Complexity science is the latest generation of systems thinking that investigates patterns and has emerged from the exploration of the subatomic world and quantum physics. A key component of complexity science is the concept of complex adaptive systems, and active research is found in many disciplines- from biology to economics to health care. However, the research and literature related to these appealing topics have generated confusion. A thorough explication of complex adaptive systems is needed. METHODS: A modified application of the methods recommended by Walker and Avant for concept analysis was used. FINDINGS: A complex adaptive system is a collection of individual agents with freedom to act in ways that are not always totally predictable and whose actions are interconnected. Examples include a colony of termites, the financial market, and a surgical team. It is often referred to as chaos theory, but the two are not the same. Chaos theory is actually a subset of complexity science. Complexity science offers a powerful new approach--beyond merely looking at clinical processes and the skills of healthcare professionals. CONCLUSION: The use of complex adaptive systems as a framework is increasing for a wide range of scientific applications, including nursing and healthcare management research. When nursing and other healthcare managers focus on increasing connections, diversity, and interactions they increase information flow and promote creative adaptation referred to as self-organization. Complexity science builds on the rich tradition in nursing that views patients and nursing care from a systems perspective. PMID- 16313379 TI - Meleis's theory of nursing transitions and relatives' experiences of nursing home entry. AB - AIM: This paper explores the extent to which Meleis's mid-range theory of nursing transitions is supported by the findings of a study exploring relatives' experiences of the move to a nursing home. BACKGROUND: Mid-range nursing theories are useful tools in helping to understand the scope of nursing practice in a range of contexts and situations. However, as yet, many formal mid-range theories have not been adequately tested. METHODS: Findings from a constructivist study of relatives' experiences of nursing home entry were re-analysed in relation to the extent to which they reflected the domains of the theory of nursing transitions. Data for the original study were generated during 37 qualitative interviews involving 48 close family members of older people who had recently moved to a nursing home, and in observational case studies in three nursing homes. FINDINGS: All domains of the theory of nursing transition were supported by the data generated within the study. However, the model failed to represent adequately the interactive and dynamic nature of relationships between formal and informal caregivers in the nursing home context. CONCLUSIONS: The theory of nursing transitions has the potential to assist nurses in identifying appropriate strategies for supporting relatives throughout the period of an older person's relocation to a nursing home. However, in order to reflect fully the experiences of relatives at this time, the theory requires adjustment to recognize the contribution made by relatives themselves to positive outcomes. This therefore raises questions as to whether the relative absence of this reciprocal and interactive dimension is an element of Meleis's theory that requires further exploration in relation to other forms of transition. PMID- 16313380 TI - The Newcastle Satisfaction with Nursing Scales: a valid measure of maternal satisfaction with inpatient postpartum nursing care. AB - AIM: To test the validity of The Newcastle Satisfaction with Nursing Scales as measures of patient satisfaction with nursing care in an inpatient postpartum unit. BACKGROUND: The Newcastle Satisfaction with Nursing Scales (Experience of Nursing Care Scale and Satisfaction with Nursing Care Scale) were developed to measure the satisfaction of medical-surgical inpatients with nursing care. METHODS: The Newcastle Satisfaction with Nursing Scales were administered by interviewers to 189 postpartum women prior to hospital discharge. We tested the construct validity of the scales by making five a priori predictions: mothers who were more satisfied would be more likely to have one nurse caring for them and to recommend the postpartum unit to a friend. We also predicted that the Experience of Nursing Care and Satisfaction with Nursing Care Scales would be positively correlated with each other, with a global question about satisfaction with nursing care, and with a global question about satisfaction with overall postpartum stay. RESULTS: Four of the five a priori predictions were supported by the data. The mean Newcastle Satisfaction with Nursing Scale scores of mothers who would recommend the unit to a friend were higher (more satisfied) than those who would not (P < 0.001). The Experience of Nursing Care Scale and Satisfaction with Nursing Care Scale were positively and significantly correlated with each other (r = 0.9, P < 0.001). There was a positive and significant correlation between the scales and global ratings of nursing care (Experience Scale r = 0.79, P < 0.001; Satisfaction Scale r = 0.82, P < 0.001) and overall postpartum stay (Experience Scale r = 0.64, P < 0.001; Satisfaction Scale r = 0.68, P < 0.001). CONCLUSION: The Newcastle Satisfaction with Nursing Scales are valid measures of maternal satisfaction with inpatient postpartum nursing care. PMID- 16313381 TI - Development of the Expectations of Filial Piety Scale--Spanish version. AB - AIMS: This paper reports the development of the Expectations of Filial Piety Scale for use with Hispanic/Latino parents regarding their expectations of care and support from their adult children. BACKGROUND: Filial piety, a term used to describe a set of family values in relation to parental care and respect, has strong cultural underpinnings. To date, there have been no measures of this construct for use with Hispanic/Latino populations. METHODS: A convenience sample of 318 Hispanic/Latino adults in North Carolina and Texas in the United States of America completed a brief biographical survey and field-tested a new Spanish version of the Expectations of Filial Piety Scale in 2003. RESULTS: Principal component factor analysis with orthogonal rotation (varimax method) was used to extract four factors, which accounted for 60% of the variance in scale scores. These factors were: I, respect for parents (23.6%); II, honouring parents (12.5%); III, supporting parents (11.9%); and IV, family unity (11.7%). Overall scale reliability was 0.82, with individual factor reliability coefficients ranging from 0.66 to 0.84. CONCLUSIONS: Increasingly, large numbers of Hispanic/Latino elders will look to their families for support as they reach advanced ages. Culturally relevant translations of family caregiving constructs, such as filial piety, will be essential nursing tools for clinical practice and in developing contemporary family caregiver research agendas. PMID- 16313382 TI - Community psychiatric nurses and the care co-ordinator role: squeezed to provide 'limited nursing'. AB - AIM: This paper reports a study illuminating the factors that either facilitate or constrain the ability of community psychiatric nurses, in their role as care co-ordinators, to meet service users' and carers' needs. BACKGROUND: The Care Programme Approach is the key policy underpinning community-focused mental health services in England, but has been unevenly implemented and is associated with increased inpatient bed use. The care co-ordinator role is central to the Care Programme Approach and is most often held by community psychiatric nurses, but there has been little research into how this role is performed or how it affects the work of community psychiatric nurses and their ability to meet the needs of service users. METHODS: A multiple case study of seven sectorised community mental health teams was employed over 2 years using predominantly qualitative methods including participant observation, semi-structured interviews and document review. The data were collected in one National Health Service trust in south England between 1999 and 2001. FINDINGS: Additional duties and responsibilities specifically associated with the care co-ordinator role and multidisciplinary working, combined with heavy workloads, produced 'limited nursing', whereby community psychiatric nurses were unable to provide evidence based psychosocial interventions that are recognized to reduce relapse amongst people with severe mental illness. CONCLUSIONS: The role of the Care Programme Approach care co-ordinator was not designed to support the provision of psychosocial interventions. Consequently, community psychiatric nurses in the co ordinator role are faced with competing demands and are unable to provide the range of structured, evidence-based interventions required. This may partially account for the increased inpatient bed use associated with the Care Programme Approach. PMID- 16313385 TI - Craniofacial development: making faces. PMID- 16313386 TI - Evolution of cranial development and the role of neural crest: insights from amphibians. AB - Contemporary studies of vertebrate cranial development document the essential role played by the embryonic neural crest as both a source of adult tissues and a locus of cranial form and patterning. Yet corresponding and basic features of cranial evolution, such as the extent of conservation vs. variation among species in the contribution of the neural crest to specific structures, remain to be adequately resolved. Investigation of these features requires comparable data from species that are both phylogenetically appropriate and taxonomically diverse. One key group are amphibians, which are uniquely able to inform our understanding of the ancestral patterns of ontogeny in fishes and tetrapods as well as the evolution of presumably derived patterns reported for amniotes. Recent data support the hypothesis that a prominent contribution of the neural crest to cranial skeletal and muscular connective tissues is a fundamental property that evolved early in vertebrate history and is retained in living forms. The contribution of the neural crest to skull bones appears to be more evolutionarily labile than that of cartilages, although significance of the limited comparative data is difficult to establish at present. Results underline the importance of accurate and reliable homology assessments for evaluating the contrasting patterns of derivation reported for the three principal tetrapod models: mouse, chicken and frog. PMID- 16313387 TI - Patterning the neural crest derivatives during development of the vertebrate head: insights from avian studies. AB - Studies carried out in the avian embryo and based on the construction of quail chick chimeras have shown that most of the skull and all the facial and visceral skeleton are derived from the cephalic neural crest (NC). Contribution of the mesoderm is limited to its occipital and (partly) to its otic domains. NC cells (NCCs) participating in membrane bones and cartilages of the vertebrate head arise from the diencephalon (posterior half only), the mesencephalon and the rhombencephalon. They can be divided into an anterior domain (extending down to r2 included) in which genes of the Hox clusters are not expressed (Hox-negative skeletogenic NC) and a posterior domain including r4 to r8 in which Hox genes of the four first paraloguous groups are expressed. The NCCs that form the facial skeleton belong exclusively to the anterior Hox-negative domain and develop from the first branchial arch (BA1). This rostral domain of the crest is designated as FSNC for facial skeletogenic neural crest. Rhombomere 3 (r3) participates modestly to both BA1 and BA2. Forced expression of Hox genes (Hoxa2, Hoxa3 and Hoxb4) in the neural fold of the anterior domain inhibits facial skeleton development. Similarly, surgical excision of these anterior Hox-negative NCCs results in the absence of facial skeleton, showing that Hox-positive NCCs cannot replace the Hox-negative domain for facial skeletogenesis. We also show that excision of the FSNC results in dramatic down-regulation of Fgf8 expression in the head, namely in ventral forebrain and in BA1 ectoderm. We have further demonstrated that exogenous FGF8 applied to the presumptive BA1 territory at the 5-6-somite stage (5-6ss) restores to a large extent facial skeleton development. The source of the cells responsible for this regeneration was shown to be r3, which is at the limit between the Hox-positive and Hox-negative domain. NCCs that respond to FGF8 by survival and proliferation are in turn necessary for the expression/maintenance of Fgf8 expression in the ectoderm. These results strongly support the emerging picture according to which the processes underlying morphogenesis of the craniofacial skeleton are regulated by epithelial mesenchymal bidirectional crosstalk. PMID- 16313388 TI - It's all in your head: new insights into craniofacial development and deformation. PMID- 16313389 TI - The role of the endoderm in the development and evolution of the pharyngeal arches. AB - The oro-pharyngeal apparatus has its origin in a series of bulges found on the lateral surface of the embryonic head, the pharyngeal arches. Significantly, the development of these structures is extremely complex, involving interactions between a number of disparate embryonic cell types: ectoderm, endoderm, mesoderm and neural crest, each of which generates particular components of the arches, and whose development must be co-ordinated to generate the functional adult oro pharyngeal apparatus. In the past most studies have emphasized the role played by the neural crest, which generates the skeletal elements of the arches, in directing pharyngeal arch development. However, it is now apparent that the pharyngeal endoderm plays an important role in directing arch development. Here we discuss the role of the pharyngeal endoderm in organizing the development of the pharyngeal arches, and the mechanisms that act to pattern the endoderm itself and those which direct its morphogenesis. Finally, we discuss the importance of modification to the pharyngeal endoderm during vertebrate evolution. In particular, we focus on the emergence of the parathyroid gland, which we have recently shown to be the result of the internalization of the gills. PMID- 16313390 TI - Developmental studies of the lamprey and hierarchical evolutionary steps towards the acquisition of the jaw. AB - The evolution of animal morphology can be understood as a series of changes in developmental programs. Among vertebrates, some developmental stages are conserved across species, representing particular developmental constraints. One of the most conserved stages is the vertebrate pharyngula, in which similar embryonic morphology is observed and the Hox code is clearly expressed. The oral developmental program also appears to be constrained to some extent, as both its morphology and the the Hox-code-default state of the oropharyngeal region are well conserved between the lamprey and gnathostome embryos. These features do not by themselves explain the evolution of jaws, but should be regarded as a prerequisite for evolutionary diversification of the mandibular arch. By comparing the pharyngula morphology of the lamprey and gnathostomes, it has become clear that the oral pattern is not entirely identical; in particular, the positional differentiation of the rostral ectomesenchyme is shifted between these animals. Therefore, the jaw seems to have arisen as an evolutionary novelty by overriding ancestral constraints, a process in which morphological homologies are partially lost. This change involves the heterotopic shift of tissue interaction, which appears to have been preceded by the transition from monorhiny to diplorhiny, as well as separation of the hypophysis. When gene expression patterns are compared between the lamprey and gnathostomes, cell-autonomously functioning genes tend to be associated with identical cell types or equivalent anatomical domains, whereas growth-factor-encoding genes have changed their expression domains during evolution. Thus, the heterotopic evolution may be based on changes in the regulation of signalling-molecule-encoding genes. PMID- 16313391 TI - Reassessing the Dlx code: the genetic regulation of branchial arch skeletal pattern and development. AB - The branchial arches are meristic vertebrate structures, being metameric both between each other within the rostrocaudal series along the ventrocephalic surface of the embryonic head and within each individual arch: thus, just as each branchial arch must acquire a unique identity along the rostrocaudal axis, each structure within the proximodistal axis of an arch must also acquire a unique identity. It is believed that regional specification of metameric structures is controlled by the nested expression of related genes resulting in a regional code, a principal that is though to be demonstrated by the regulation of rostrocaudal axis development in animals exerted by the nested HOM-C/Hox homeobox genes. The nested expression pattern of the Dlx genes within the murine branchial arch ectomesenchyme has more recently led to the proposal of a Dlx code for the regional specification along the proximodistal axis of the branchial arches (i.e. it establishes intra-arch identity). This review re-examines this hypothesis, and presents new work on an allelic series of Dlx loss-of-function mouse mutants that includes various combinations of Dlx1, Dlx2, Dlx3, Dlx5 and Dlx6. Although we confirm fundamental aspects of the hypothesis, we further report a number of novel findings. First, contrary to initial reports, Dlx1, Dlx2 and Dlx1/2 heterozygotes exhibit alterations of branchial arch structures and Dlx2-/- and Dlx1/2-/- mutants have slight alterations of structures derived from the distal portions of their branchial arches. Second, we present evidence for a role for murine Dlx3 in the development of the branchial arches. Third, analysis of compound Dlx mutants reveals four grades of mandibular arch transformations and that the genetic interactions of cis first-order (e.g. Dlx5 and Dlx6), trans second-order (e.g. Dlx5 and Dlx2) and trans third-order paralogues (e.g. Dlx5 and Dlx1) result in significant and distinct morphological differences in mandibular arch development. We conclude by integrating functions of the Dlx genes within the context of a hypothesized general mechanism for the establishment of pattern and polarity in the first branchial arch of gnathostomes that includes regionally secreted growth factors such as Fgf8 and Bmp and other transcription factors such as Msx1, and is consistent both with the structure of the conserved gnathostome jaw bauplan and the elaboration of this bauplan to meet organismal end-point designs. PMID- 16313392 TI - Developmental mechanisms facilitating the evolution of bills and quills. AB - Beaks and feathers epitomize inimitable avian traits. Within individuals and across species there exists astounding diversity in the size, shape, arrangement, and colour of beaks and feathers in association with various functional adaptations. What has enabled the concomitantly divergent evolution of beaks and feathers? The common denominator may lie in their developmental programmes. As revealed through recent transplant experiments using quail and duck embryos, the developmental programme for each structure utilizes mesenchyme as a dominant source of species-specific patterning information, acts as a module of closely coupled molecular and histogenic events, and operates with a high degree of spatial and temporal plasticity. By synergizing these three features, the developmental programmes underlying beaks and feathers likely have the essential potential to react spontaneously to novel conditions and new gene functions, and as a consequence are well equipped to generate and accommodate innovative phenotypes during the course of evolution. PMID- 16313394 TI - Deer antlers: a zoological curiosity or the key to understanding organ regeneration in mammals? AB - Many organisms are able to regenerate lost or damaged body parts that are structural and functional replicates of the original. Eventually these become fully integrated into pre-existing tissues. However, with the exception of deer, mammals have lost this ability. Each spring deer shed antlers that were used for fighting and display during the previous mating season. Their loss is triggered by a fall in circulating testosterone levels, a hormonal change that is linked to an increase in day length. A complex 'blastema-like' structure or 'antler-bud' then forms; however, unlike the regenerative process in the newt, most evidence (albeit indirect) suggests that this does not involve reversal of the differentiated state but is stem cell based. The subsequent re-growth of antlers during the spring and summer months is spectacular and represents one of the fastest rates of organogenesis in the animal kingdom. Longitudinal growth involves endochondral ossification in the tip of each antler branch and bone growth around the antler shaft is by intramembranous ossification. As androgen concentrations rise in late summer, longitudinal growth stops, the skin (velvet) covering the antler is lost and antlers are 'polished' in preparation for the mating season. Although the timing of the antler growth cycle is clearly closely linked to circulating testosterone, oestrogen may be a key cellular regulator, as it is in the skeleton of other male mammals. We still know very little about the molecular machinery required for antler regeneration, although there is evidence that developmental signalling pathways with pleiotropic functions are important and that novel 'antler-specific' molecules may not exist. Identifying these pathways and factors, deciphering their interactions and how they are regulated by environmental cues could have an important impact on human health if this knowledge is applied to the engineering of new human tissues and organs. PMID- 16313395 TI - Barx1 and evolutionary changes in feeding. AB - During mouse embryonic development, the Barx1 homeobox gene is expressed in the mesenchymal cells of molar teeth and stomach. During early stages of molar development, Barx1 has an instructive role, directing the as yet undetermined ectomesenchymal cells in the proximal region of the jaws to follow a multicuspid tooth developmental pathway. We review here recent results showing an absence of stomach tissue in Barx1 mutant mice. The data strongly suggest that in the presumptive stomach mesenchyme Barx1 acts to attenuate Wnt signalling allowing digestive tract endoderm to differentiate into a highly specialized stomach epithelium. In the light of these new data, we discuss the possibility that evolutionary changes in the Barx1 gene could have simultaneously altered the dentition and the digestive system, therefore positioning Barx1 as a key gene in the evolution of mammals. PMID- 16313393 TI - Relations and interactions between cranial mesoderm and neural crest populations. AB - The embryonic head is populated by two robust mesenchymal populations, paraxial mesoderm and neural crest cells. Although the developmental histories of each are distinct and separate, they quickly establish intimate relations that are variably important for the histogenesis and morphogenesis of musculoskeletal components of the calvaria, midface and branchial regions. This review will focus first on the genesis and organization within nascent mesodermal and crest populations, emphasizing interactions that probably initiate or augment the establishment of lineages within each. The principal goal is an analysis of the interactions between crest and mesoderm populations, from their first contacts through their concerted movements into peripheral domains, particularly the branchial arches, and continuing to stages at which both the differentiation and the integrated three-dimensional assembly of vascular, connective and muscular tissues is evident. Current views on unresolved or contentious issues, including the relevance of head somitomeres, the processes by which crest cells change locations and constancy of cell-cell relations at the crest-mesoderm interface, are addressed. PMID- 16313396 TI - Neurulation in the cranial region--normal and abnormal. AB - Cranial neurulation is the embryonic process responsible for formation of the brain primordium. In the mouse embryo, cranial neurulation is a piecemeal process with several initiation sites and two neuropores. Variation in the pattern of cranial neurulation occurs in different mouse strains, and a simpler version of this morphogenetic scheme has been described in human embryos. Exencephaly is more common in females than in males, an unexplained phenomenon seen in both mice and humans. As the cranial neural tube closes, a critical morphogenetic event is the formation of dorsolateral bending points near the neural fold tips, which enables subsequent midline fusion of the neural folds. Many mutant and gene targeted mouse strains develop cranial neural tube defects, and analysis of the underlying molecular defects identifies several requirements for normal dorsolateral bending. These include a functional actin cytoskeleton, emigration of the cranial neural crest, spatio-temporally regulated apoptosis, and a balance between cell proliferation and the onset of neuronal differentiation. A small number of mouse mutants exhibit craniorachischisis, a combined brain and spine neurulation defect. Recent studies show that disturbance of a single molecular signalling cascade, the planar cell polarity pathway, is implicated in mutants with this defect. PMID- 16313397 TI - Growth of the normal skull vault and its alteration in craniosynostosis: insights from human genetics and experimental studies. AB - The mammalian skull vault is constructed principally from five bones: the paired frontals and parietals, and the unpaired interparietal. These bones abut at sutures, where most growth of the skull vault takes place. Sutural growth involves maintenance of a population of proliferating osteoprogenitor cells which differentiate into bone matrix-secreting osteoblasts. Sustained function of the sutures as growth centres is essential for continuous expansion of the skull vault to accommodate the growing brain. Craniosynostosis, the premature fusion of the cranial sutures, occurs in 1 in 2500 children and often presents challenging clinical problems. Until a dozen years ago, little was known about the causes of craniosynostosis but the discovery of mutations in the MSX2, FGFR1, FGFR2, FGFR3, TWIST1 and EFNB1 genes in both syndromic and non-syndromic cases has led to considerable insights into the aetiology, classification and developmental pathology of these disorders. Investigations of the biological roles of these genes in cranial development and growth have been carried out in normal and mutant mice, elucidating their individual and interdependent roles in normal sutures and in sutures undergoing synostosis. Mouse studies have also revealed a significant correspondence between the neural crest-mesoderm boundary in the early embryonic head and the position of cranial sutures, suggesting roles for tissue interaction in suture formation, including initiation of the signalling system that characterizes the functionally active suture. PMID- 16313398 TI - Regional regulation of palatal growth and patterning along the anterior-posterior axis in mice. AB - Cleft palate is a congenital disorder arising from a failure in the multistep process of palate development. In its mildest form the cleft affects only the posterior soft palate. In more severe cases the cleft includes the soft (posterior) and hard (anterior) palate. In mice a number of genes show differential expression along the anterior-posterior axis of the palate. Mesenchymal heterogeneity is established early, as evident from Bmp4-mediated induction of Msx1 and cell proliferation exclusively in the anterior and Fgf8 specific induction of Pax9 in the posterior palate alone. In addition, the anterior palatal epithelium has the unique ability to induce Shox2 expression in the anterior mesenchyme in vivo and the posterior mesenchyme in vitro. Therefore, the induction and competence potentials of the epithelium and mesenchyme in the anterior are clearly distinct from those in the posterior. Defective growth in the anterior palate of Msx1-/- and Fgf10-/- mice leads to a complete cleft palate and supports the anterior-to-posterior direction of palatal closure. By contrast, the Shox2-/- mice exhibit incomplete clefts in the anterior presumptive hard palate with an intact posterior palate. This phenotype cannot be explained by the prevailing model of palatal closure. The ability of the posterior palate to fuse independent of the anterior palate in Shox2-/- mice underscores the intrinsic differences along the anterior-posterior axis of the palate. We must hitherto consider the heterogeneity of gene expression and function in the palate to understand better the aetiology and pathogenesis of non-syndromic cleft palate and the mechanics of normal palatogenesis. PMID- 16313399 TI - Three-dimensional analysis of mandibular growth and tooth eruption. AB - Normal and abnormal jaw growth and tooth eruption are topics of great importance for several dental and medical disciplines. Thus far, clinical studies on these topics have used two-dimensional (2D) radiographic techniques. The purpose of the present study was to analyse normal mandibular growth and tooth eruption in three dimensions based on computer tomography (CT) scans, extending the principles of mandibular growth analysis proposed by Bjork in 1969 from two to three dimensions. As longitudinal CT data from normal children are not available (for ethical reasons), CT data from children with Apert syndrome were employed, because it has been shown that the mandible in Apert syndrome is unaffected by the malformation, and these children often have several craniofacial CT scans performed during childhood for planning of cranial and midface surgery and for follow-up after surgery. A total of 49 datasets from ten children with Apert syndrome were available for study. The number of datasets from each individual ranged from three to seven. The first CT scan in each of the ten series was carried out before 1 year of age, and the ages for the 49 scans ranged from 1 week to 14.5 years. The mandible and the teeth were segmented and iso-surfaces generated. Landmarks were placed on the surface of the mandible, along the mandibular canals, the inner contour of the cortical plate at the lower border of the symphysis menti, and on the teeth. Superimposition of the mandibles in the longitudinal series was performed using the symphysis menti and the mandibular canals as suggested by Bjork. The study supported the findings of stability of the symphysis menti and the mandibular canals as seen in profile view previously reported by Bjork & Skieller in 1983. However, the mandibular canals were, actually, relocated laterally during growth. Furthermore, the position of tooth buds remained relatively stable inside the jaw until root formation started. Eruption paths of canines and premolars were vertical, whereas molars erupted in a lingual direction. The 3D method would seem to offer new insight into jaw growth and tooth eruption, but further studies are needed. PMID- 16313400 TI - Antibiotic resistance in staphylococci associated with cats and dogs. PMID- 16313401 TI - Suitability of bifidobacteria and thymol as biopreservatives in extending the shelf life of fresh packed plaice fillets. AB - AIMS: The present study was aimed to evaluate the microbiological and sensory characteristics of biopreserved packed fresh plaice (Pleuronectes platessa), during storage. METHODS AND RESULTS: Fish samples were treated with a Bifidobacterium bifidum strain and thymol as preservatives, stored at different temperatures (4 and 12 degrees C) and package atmospheres (in air, under vacuum, modified atmosphere). Data about the total viable count (TVC) were used to predict the shelf life of the fish fillets as affected by the preservatives used and the storage conditions. Data obtained revealed that B. bifidum could perform an efficient synergy with thymol, in controlling the hygiene biological markers. Besides, the tested bifidobacterial species, combined with low storage temperature and anoxia/hypoxia, showed a great efficacy against the main fresh packaged fish spoilage species (Pseudomonas spp., Photobacterium phosphoreum). CONCLUSIONS: The studied technique could be considered effective in fresh fish spoilage control. SIGNIFICANCE AND IMPACT OF THE STUDY: Great interest is developing on food biopreservation, because of the ever increasing needs to protect consumers' and environment's health, and to revalorize some important food characteristics, such as naturalness and authenticity, without leaving the safety aside. PMID- 16313402 TI - Effect of Lactobacillus-protective cultures with bacteriocin-like inhibitory substances' producing ability on microbiological, chemical and sensory changes during storage of refrigerated vacuum-packaged sliced beef. AB - AIMS: To investigate the effect of applying two different Lactobacillus protective cultures, with bacteriocin-like inhibitory substances' (BLIS) producing ability, individually or in combination, on microbiological, chemical and sensory changes during storage of refrigerated vacuum-packaged sliced beef meat. METHODS AND RESULTS: Lactobacillus sakei CECT 4808 and Lactobacillus curvatus CECT 904(T), which were shown to be producers of BLIS, were inoculated individually or in combination on slices of beef M. semitendinosus. The samples were vacuum packaged and stored at 4 +/- 1 degrees C and were assessed during a 28-day storage period for microbiological [Enterobacteriaceae, Pseudomonas spp., lactic acid bacteria (LAB), Brochothrix thermosphacta and yeasts and moulds], chemical (pH, protein hydrolysis degree, lipid oxidation), sensory (abnormal odour) parameters and instrumental colour. Samples inoculated with the Lact. sakei strain and samples inoculated with the combination of the two strains had significantly (P < 0.05) lower spoilage microbial counts than those inoculated with the Lact. curvatus strain alone or the controls, while both chemical parameters (including lipid oxidation) and abnormal odour scores were also significantly (P < 0.05) improved by the former. Moreover, Lact. sakei alone showed a better preserving effect (P < 0.05) than the combination of both strains in the majority of the parameters tested. Instrumental colour measurements changed with storage time, but no treatment effects (P >or= 0.05) were observed during the whole 28-day storage period. CONCLUSIONS: The BLIS producer Lact. sakei CECT 4808 strain may be used for improving preservation of vacuum-packaged beef slices, as regards spoilage microbial counts and the chemical parameters tested in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: Inoculation with the BLIS producer Lact. sakei CECT 4808 strain would provide an additional hurdle to improve storage life of refrigerated vacuum-packaged sliced beef. Furthermore, this strain demonstrated limited antioxidative ability, which could make a contribution to the prevention of lipid oxidation in meat and meat products. PMID- 16313403 TI - Growth and bacteriocin production kinetics of Leuconostoc mesenteroides E131. AB - AIMS: The aim of this study was to investigate the effect of pH, temperature, sodium chloride, type and level of sugar used in fermented sausages, on the occurrence and the concentration of the maximum bacteriocin activity, in order to optimize the bacteriocin synthesis during the growth cycle of Leuconostoc mesenteroides E131. METHODS AND RESULTS: In order to study the effect of the environmental factors on growth and bacteriocin production of Leuc. mesenteroides E131 fermentations were carried out in fermentor as well as in flasks. Mathematical equations were used to describe the kinetic parameters of the strain. When the micro-organism was grown in lower pH value (5.5) than optimum for growth (pH 6.5) the bacteriocin production was enhanced. On the contrary, bacteriocin production was favoured when the micro-organism was grown at temperatures close to the optimum for growth (25 degrees C). Finally, the level and the type of the sugar used as carbon source affected both growth and bacteriocin production with glucose being better source for biomass production and fructose more suitable for bacteriocin production. CONCLUSIONS: Leuconostoc mesenteroides E131 has a potential use as protective culture or its bacteriocin as protective agent in combination with another starter culture in fermented meats. SIGNIFICANCE AND IMPACT OF THE STUDY: Better understanding of the influence of environmental factors, such as pH, temperature and carbon source on the kinetic behaviour of Leuc. mesenteroides E131. PMID- 16313404 TI - Interaction of air ions and bactericidal vapours to control micro-organisms. AB - AIMS: The aim of this study was to investigate the antibacterial activity of candles containing specific-antibacterial compounds, such as essential oils and their constituent compounds. The importance of the ionization products from the flame and the aerial concentration of the volatile compounds were investigated. METHODS AND RESULTS: Agar plates inoculated with Escherichia coli (DH5alpha) or Staphylococcus aureus (NCTC strain number 8532) were exposed in a large air-tight chamber to candle flames combined with the volatile bactericidal compounds beta pinene and orange oil. A steady decline in E. coli numbers was observed over time because of the effect of a candle flame. This was significantly increased by the addition of volatile oils. The number of S. aureus colonies was not reduced by a plain candle, but significant reductions were caused following exposure to beta pinene and orange oil candles. As aerial concentration of the volatiles was increased the viability of E. coli and S. aureus declined. Ionization products from the flame made a significant contribution to the observed effects, as intercepting the ions on a grounded grid over the agar plates allowed at least 20% more cells to survive. CONCLUSIONS: This study demonstrates the antibacterial properties of ionization products from a candle flame, and that this effect can be significantly increased by the addition of specific-antibacterial compounds, such as orange oil and beta-pinene. The role of both the ionization products from the candle flame and the concentration of volatile compounds released are important to the effect. SIGNIFICANCE AND IMPACT OF THE STUDY: The technique described here offers a new and novel technique for reducing the concentration of bacteria on surfaces. PMID- 16313405 TI - Influence of fermentation time, cryoprotectant and neutralization of cell concentrate on freeze-drying survival, storage stability, and acid and bile exposure of Bifidobacterium animalis ssp. lactis cells produced without milk based ingredients. AB - AIMS: To investigate the stability of Bifidobacterium animalis ssp. lactis VTT E 012010 (=Bb-12) during freeze-drying, storage and acid and bile exposure. The effect of harvesting time and composition and pH of the cryoprotectant on the survival was evaluated. The procedure was performed by using a milk-free culture medium and cryoprotectants to produce cells for nonmilk-based applications. METHODS AND RESULTS: Bifidobacterial cells were grown in fermenters in general edible medium for 15 or 22 h. The cell mass was freeze-dried either as non neutralized or neutralized using sucrose, betaine or reconstituted skim milk (control) as cryoprotectants. For stability studies freeze-dried powders were stored at 37, 5 and -20 degrees C for 2-6 months. In addition, acid and bile tolerance of the powders was tested. Sucrose-formulated B. animalis ssp. lactis preparations had an excellent stability during storage at refrigerated and frozen temperatures for 5-6 months. They also had a good survival during storage at 37 degrees C for 2 months as well as during exposure to pH 3 and 1% bile acids. No difference was observed between 15 and 22 h grown cells or between non neutralized and neutralized cells. Betaine proved to be a poor cryoprotectant compared with sucrose. CONCLUSIONS: Fermentation time and neutralization of cell concentrate before freeze-drying had no impact on the storage stability and bile and acid tolerance of freeze-dried bifidobacterial cells. The nonmilk-based production protocol using sucrose as a cryoprotectant yielded powdery preparations with excellent stability in adverse conditions (storage at elevated temperatures and during acid and bile exposure). SIGNIFICANCE AND IMPACT OF THE STUDY: The results indicate that it is feasible to develop nonmilk-based production technologies for probiotic cultures. This provides new possibilities for the development of nondairy-based probiotic products. PMID- 16313406 TI - Evaluation of chromogenic media for the detection of Listeria species in food. AB - AIMS: The aim of this study was to evaluate the performance of chromogenic agars, Agar Listeria according to Ottaviani and Agosti (ALOA) and Rapid L. mono agar, compared with Oxford agar for the enumeration and detection of Listeria species in food. METHODS AND RESULTS: A total of 170 food samples were examined using the three plating media. Listeria species were isolated from 63 samples. In contrast to Oxford agar, detection of Listeria colonies on chromogenic media was as good after 24 h of incubation of plates as after 48 h. While there was no significant difference in recovery of Listeria monocytogenes on the three media, recovery of other Listeria species was significantly poorer on Rapid L. mono agar compared with Oxford and ALOA agars. Recovery of species other than L. monocytogenes was significantly improved by including a secondary enrichment stage in the detection method. CONCLUSIONS: Using chromogenic agars, presumptive identification of L. monocytogenes is possible after 24 h, compared with 3-4 days using Oxford agar. However, the poor detection of species other than L. monocytogenes on Rapid L. mono agar is a disadvantage of this medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new information regarding the isolation of Listeria species other than L. monocytogenes from food using chromogenic plating media. This is important, as non-pathogenic Listeria species act as markers for the likelihood of presence of L. monocytogenes and allow preventive action to be taken to avoid its presence. PMID- 16313407 TI - In vitro pharmacodynamic evaluation of antiviral medicinal plants using a vector based assay technique. AB - AIMS: Medicinal plants are increasingly being projected as suitable alternative sources of antiviral agents. The development of a suitable in vitro pharmacodynamic screening technique could contribute to rapid identification of potential bioactive plants and also to the standardization and/or pharmacokinetic pharmacodynamic profiling of the bioactive components. METHODS AND RESULTS: Recombinant viral vectors (lentiviral, retroviral and adenoviral) transferring the firefly luciferase gene were constructed and the inhibition of viral vector infectivity by various concentrations of plant extracts was evaluated in HeLa or Hep2 cells by measuring the changes in luciferase activity. Cytotoxicity of the extracts was evaluated in parallel on HeLa or Hep2 cells stably expressing luciferase. Amongst the 15 extracts screened, only the methanol (ME) and the ethyl acetate (ET) fractions of the lichen, Ramalina farinacea specifically reduced lentiviral and adenoviral infectivity in a dose-dependent manner. Further, chromatographic fractionation of ET into four fractions (ET1-ET4) revealed only ET4 to be selectively antiviral with an IC50 in the 20 microg ml( 1) range. Preliminary mechanistic studies based on the addition of the extracts at different time points in the viral infection cycle (kinetic studies) revealed that the inhibitory activity was highest if extract and vectors were preincubated prior to infection, suggesting that early steps in the lentiviral or adenoviral replication cycle could be the major target of ET4. Inhibition of wild-type HIV-1 was also observed at a 10-fold lower concentration of the extract. CONCLUSIONS: The vector-based assay is a suitable in vitro pharmacodynamic evaluation technique for antiviral medicinal plants. The technique has successfully demonstrated the presence of antiviral principles in R. farinacea. SIGNIFICANCE AND IMPACT OF STUDY: Potential anti-HIV medicinal plants could rapidly be evaluated with the reported vector-based technique. The lichen, R. farinacea could represent a lead source of antiviral substances and is thus worthy of further studies. PMID- 16313408 TI - A survey of ammonia-assimilating micro-organisms in cattle manure composting. AB - AIMS: To evaluate the ammonia-assimilating abilities of micro-organisms isolated from cattle manure composting processes and to determine the distribution of cultivable species of ammonia-assimilating micro-organisms in microbial communities during the composting processes. METHODS AND RESULTS: Compost samples were collected from four stages of treatment. Trypto soya agar was used for the isolation of ammonia-assimilating aerobes. Many of the isolates showed high ammonia-assimilating ability in a medium containing basal components and a compost extract. Partial 16S ribosomal DNA sequencing showed that the cultivable species of highly efficient ammonia-assimilating isolates changed during the composting process. The community structure of micro-organisms and actinomycetes was analysed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Two species of actinomycetes identified by PCR-DGGE coincided with those found among the cultured isolates. CONCLUSIONS: Ammonia-assimilating micro organisms obtained by the cultivation method were not predominant in the microbial community during the composting process: however certain cultured actinomycetes were members of predominant species in the actinomycetes community. SIGNIFICANCE AND IMPACT OF THE STUDY: Ammonia assimilation by micro-organisms is one of the important mechanisms for ammonia retention in the composting process. Cultivable actinomycetes are a means for preventing ammonia emission from the composting process. PMID- 16313409 TI - Synergistic effect of enterocin AS-48 in combination with outer membrane permeabilizing treatments against Escherichia coli O157:H7. AB - AIMS: To determine the effects of outer membrane (OM) permeabilizing agents on the antimicrobial activity of enterocin AS-48 against Escherichia coli O157:H7 CECT 4783 strain in buffer and apple juice. METHODS AND RESULTS: We determined the influence of pH, EDTA, sodium tripolyphosphate (STPP) and heat on E. coli O157:H7 CECT 4783 sensitivity to enterocin AS-48 in buffer and in apple juice. Enterocin AS-48 was not active against intact cells of E. coli O157:H7 CECT 4783 at neutral pH. However, cells sublethally injured by OM permeabilizing agents (EDTA, STPP, pH 5, pH 8.6 and heat) became sensitive to AS-48, decreasing the amount of bacteriocin required for inhibition of E. coli O157:H7 CECT 4783. CONCLUSIONS: The results presented indicate that enterocin AS-48 could potentially be applied with a considerably wider range of protective agents, such as OM permeabilizing agents, with increased efficacy in inhibiting E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: Results from this study support the potential use of enterocin AS-48 to control E. coli O157:H7 in combination with other hurdles. PMID- 16313410 TI - Cytotoxicity and apoptosis induction of some selected marine bacteria metabolites. AB - AIMS: To study the potential apoptosis effects of cytotoxic marine bacterial metabolites on human HeLa cell line. METHODS AND RESULTS: After HeLa cells were routinely cultured, tetrazolium-based colorimetric assay for cytotoxicity was performed to screen the marine bacteria extracts showing 12 strains active. To find the potential active strain with apoptosis mechanism, a battery of apoptosis assays, including AO/EB staining, TUNEL assay (terminal-deoxynucleotidyl transferase mediated nick end labelling), gel electrophoresis and flow cytometry, were used to determine whether apoptosis was involved in HeLa cell cytotoxicity of marine bacterial extracts. The results indicated that four strains could induce cell shrinkage, cell membrane blebbing, formation of apoptotic body and DNA fragmentation. CONCLUSIONS: Crude extracts of 12 of 153 strains of marine bacteria showed cytotoxic effects with ID50 ranged from 77.20 to 199.84 microg ml(-1), in which eight strains of bacteria were associated bacteria. The metabolites in the strains of QD1-2, NJ6-3-1, NJ1-1-1 and SS6-4 were able to induce HeLa cells apoptosis. Furthermore, the assessment by flow cytometry indicated that the hypodiploid apoptotic cells increased in a time-dependent manner, suggesting that induced apoptosis occurred from 24 h to 48 h after the extracts treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggested that the compounds from fermentation in these four marine bacterial strains could be candidates for developing apoptosis specific anti-tumour agents with lower toxicity. This study indicated that associated marine bacteria could be good source to find cytotoxic metabolites, and some cytotoxic marine bacterial metabolites could have apoptosis mechanisms. PMID- 16313411 TI - Effects of culture conditions on the growth and auto-aggregation ability of vaginal Lactobacillus johnsonii CRL 1294. AB - AIMS: To evaluate the effects of different physico-chemical factors on the growth and auto-aggregating ability of vaginal Lactobacillus johnsonii CRL 1294. METHODS AND RESULTS: L. johnsonii CRL 1294 was cultivated in different culture media, initial pH and temperature of incubation. The growth parameters were estimated by the Gompertz model, being optimal (higher final biomass and growth rate, and shorter lag phase) at an initial pH of 6.5 and at a temperature of 37 degrees C, both in LAPTg and MRS. The auto-aggregation ability, which was assessed by a model of exponential association, was evidenced in all the growth phases, being higher at pH 5 or 6.5. CONCLUSIONS: The growth of L. johnsonii CRL 1294 was affected in different way by all the physico-chemical factors tested. However, the auto-aggregation ability increased mainly at low initial pH of growth media. SIGNIFICANCE AND IMPACT OF THE STUDY: The auto-aggregation ability under different culture conditions of a vaginal Lactobacillus strain was systematically and statistically evaluated for the first time. The higher cellular aggregation evidenced at low pH could be a fundamental characteristic in the acidic vaginal environment to promote the protective role of lactobacilli. PMID- 16313412 TI - Assessing the role of alkane hydroxylase genotypes in environmental samples by competitive PCR. AB - AIMS: A molecular tool for extensive detection of prokaryotic alkane hydroxylase genes (alkB) was developed. AlkB genotypes involved in the degradation of short chain alkanes were quantified in environmental samples in order to assess their occurrence and ecological importance. METHODS AND RESULTS: Four primer pairs specific for distinct clusters of alkane hydroxylase genes were designed, allowing amplification of alkB-related genes from all tested alkane-degrading strains and from six of seven microcosms. For the primer pair detecting alkB genes related to the Pseudomonas putida GPo1 alkB gene and the one targeting alkB genes of Gram-positive strains, both involved in short-chain alkane degradation (90%) throughout the experiment. Specific methanogenic activity measurements of biomass from both reactors indicated that the main route of methane production was hydrogenotrophic methanogenesis. Terminal restriction fragment length polymorphism (TRFLP) analysis was applied to the characterization of microbial community dynamics within the system during the trial. The principal differences between the two consortia analysed included an increased abundance of Thiovulum- and Methanococcus-like organisms and uncultured Crenarchaeota in R1. CONCLUSIONS: The results indicated that there was a microbiological basis for the deviation, in terms of operational performance, of R1 and R2. SIGNIFICANCE AND IMPACT OF THE STUDY: High-throughput fingerprinting techniques, such as TRFLP, have been demonstrated as practically relevant for biomonitoring of anaerobic reactor communities. PMID- 16313420 TI - Pulsed-field gel electrophoresis, plasmid profiles and phage types for the human isolates of Salmonella enterica serovar Enteritidis obtained over 13 years in Taiwan. AB - AIMS: Plasmid profile, phage typing, and pulsed-field gel electrophoresis (PFGE) patterns of 124 Salmonella Enteritidis strains isolated in 1998-2002 in Taiwan were analysed and the results were compared with those of the 63 strains obtained in 1991-1997, so that molecular subtypes and epidemic strains for Salmonella Enteritidis over a 13-year period (1991-2002) could be elucidated. METHODS AND RESULTS: A total of 124 strains of Salmonella Enteritidis isolated from human in Taiwan between 1998 and 2002 were analysed by PFGE, plasmid analysis and phage typing. The results obtained were compared with those of the 63 strains obtained in 1991-1997, so that the clonal relationships for a total of 187 strains obtained over 13 years could be elucidated. For PFGE, restriction enzymes XbaI, SpeI and NotI were used for chromosomal DNA digestion. Results showed 28 PFGE pattern combinations for the 187 Salmonella strains. Of them, pattern X3S3N3 was the major subtype as 130 strains isolated from different locations during 1991 2002 showed this PFGE pattern. For all these 187 strains, the genetic similarity was higher than 80%. Plasmid analysis showed 17 distinct types, which consist of one to four plasmids and the predominant phage type of those strains was PT4 (71.6%) and PT6a (13.4%). The three methods identified different degrees of polymorphism in the following order: plasmid profile (18 types, D = 0.659) > PFGE (28 types, D = 0.512) > phage typing (13 types, D = 0.438). As PFGE patterns, phage type and plasmid profile were combined for subtyping, the 187 strains could be grouped into 46 subtypes and the discriminatory index was raised to 0.795. For these 46 subtypes, the predominant one was X3S3N3/P1/PT4, which contained 77 (41%) isolates. CONCLUSIONS: Most of the Salmonella Enteritidis strains from sporadic cases were with pattern X3S3N3. They were the prevalent and may be the epidemic strains found in Taiwan during 1991-2002. The present study suggested that the several variants were derived from a single clonal line and the genome for strains of Salmonella Enteritidis are highly conserved over a 13-year period (1991-2002). SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained here are useful for epidemiolgical study of salmonellosis caused by Salmonella Enteritidis in Taiwan. Comparing the data of the present study with those obtained for strains from other countries, the major subtypes for Salmonella Enteritidis infection in the world can be elucidated. PMID- 16313421 TI - Effect of mechanical abrasion on the viability, disruption and germination of spores of Bacillus subtilis. AB - AIMS: To elucidate the factors influencing the sensitivity of Bacillus subtilis spores in killing and disrupting by mechanical abrasion, and the mechanism of stimulation of spore germination by abrasion. METHODS AND RESULTS: Spores of B. subtilis strains were abraded by shaking with glass beads in liquid or the dry state, and spore killing, disruption and germination were determined. Dormant spores were more resistant to killing and disruption by abrasion than were growing cells or germinated spores. However, dormant spores of the wild-type strain with or without most coat proteins removed, spores of strains with mutations causing spore coat defects, spores lacking their large depot of dipicolinic acid (DPA) and spores with defects in the germination process exhibited essentially identical rates of killing and disruption by abrasion. When spores lacking all nutrient germinant receptors were enumerated by plating directly on nutrient medium, abrasion increased the plating efficiency of these spores before killing them. Spores lacking all nutrient receptors and either of the two redundant cortex-lytic enzymes behaved similarly in this regard, but the plating efficiency of spores lacking both cortex-lytic enzymes was not stimulated by abrasion. CONCLUSIONS: Dormant spores are more resistant to killing and disruption by abrasion than are growing cells or germinated spores, and neither the complete coats nor DPA are important in spore resistance to such treatments. Germination is not essential for spore killing by abrasion, although abrasion can trigger spore germination by activation of either of the spore's cortex-lytic enzymes. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides new insight into the mechanisms of the killing, disruption and germination of spores by abrasion and makes the surprising finding that at least much of the spore coat is not important in spore resistance to abrasion. PMID- 16313422 TI - Proteolytic activity and reduction of gliadin-like fractions by sourdough lactobacilli. AB - AIMS: To characterize the peptide hydrolase system of Lactobacillus plantarum CRL 759 and CRL 778 and evaluate their proteolytic activity in reducing gliadin-like fractions. METHODS AND RESULTS: The intracellular peptide hydrolase system of Lact. plantarum CRL 759 and CRL 778 involves amino-, di- (DP), tri- (TP) and endopeptidase activities. These peptidases are metalloenzymes inhibited by EDTA and 1,10-phenanthroline and stimulated by Co2+. DP and TP activities of Lact. plantarum CRL 759 and CRL 778, respectively, were completely inhibited by Cu2+. Lactobacillus plantarum CRL 778 showed the highest proteolytic activity and amino acids release in fermented dough. The synthetic 31-43 alpha-gliadin fragment was hydrolysed to 36% and 73% by Lact. plantarum CRL 778 and CRL 759 respectively. CONCLUSIONS: Lactobacillus plantarum CRL 759 and CRL 778 have an active proteolytic system, which is responsible for the high amino acid release during sourdough fermentation and the hydrolysis of the 31-43 alpha-gliadin-like fragment. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides new information of use when obtaining sourdough starters for bread making. Moreover, knowledge regarding lactobacilli capable of reducing the level of gliadin-like fractions, a toxic peptide for coeliac patients, has a beneficial health impact. PMID- 16313423 TI - Environmental factors influencing the relationship between optical density and cell count for Listeria monocytogenes. AB - AIMS: The effect of temperature (2-30 degrees C), pH (4.8-7.4) and water activity (0.946-0.995) on the relationship between optical density (OD) at 600 nm and the plate count (CFU ml(-1)) was investigated for Listeria monocytogenes. METHODS AND RESULTS: Calibration curves, relating OD with plate counts, were collected by measuring the OD of consecutive one-half dilution series, before determining the cell density by classic plate count methods. The calibration curves were observed to be shifting in a parallel way, with increasing stress levels. Especially pH influenced the curve in a great extent, while the other variables were showing more synergetic effects. The reason for the shift was investigated by a microscopic viability test, showing a viability decrease with increasing stress levels, causing the shift of the calibration curve. In a last step a model was made describing the effect of environmental factors on the calibration curve, with different data transformations being tested. A polynomial equation was fitted to the data, taking into account a set of constraints to incorporate microbiological knowledge in the black box model. Hence, illogical interpolation results and overfitting of the data could be avoided. CONCLUSIONS: Different stress factors are affecting the relationship between the OD and the cell count of L. monocytogenes by lowering the cell viability. These effects could be modelled using a constrained polynomial model. SIGNIFICANCE AND IMPACT OF THE STUDY: The observed phenomena are important when calculating growth parameters, like growth rate and lag phase, based on OD data. PMID- 16313424 TI - Anti-bacterial activity of Lactobacillus plantarum strain SK1 against Listeria monocytogenes is due to lactic acid production. AB - AIMS: The aim of this research was to investigate the potential of Lactobacillus plantarum strain SK1 for use as a biological control agent against Listeria monocytogenes and determine its mechanism of anti-listerial activity. METHODS AND RESULTS: Co-growth of Lact. plantarum SK1 and L. monocytogenes UMCC98 in MRS broth showed that anti-listerial activity of Lact. plantarum SK1 occurred during late log/early stationary phase of growth. This coincided with a reduction in broth pH to 4.26. Evidence obtained from the analysis of cell-free culture filtrates of strain SK1 grown in MRS broth using thin-layer chromatography and growth of L. monocytogenes in pH-adjusted culture filtrates suggested that the anti-listerial activity was due to lactic acid production alone. Trials of Lact. plantarum SK1 on radishes stored at 5 degrees C showed that it had statistically significant (P < 0.05) anti-listerial activity. CONCLUSIONS: The anti-listerial activity of Lact. plantarum SK1 was due to lactic acid production alone. A small scale trial on radishes stored at 5 degrees C showed it to have significant anti listerial activity in planta. SIGNIFICANCE AND IMPACT OF THE STUDY: This organism has potential as a biological control agent for L. monocytogenes. PMID- 16313426 TI - Annotation: what do we know about sensory dysfunction in autism? A critical review of the empirical evidence. AB - BACKGROUND: Unusual responses to sensory stimuli are seen in many children with autism. Their presence was highlighted both in early accounts of autism and in more recent first-person descriptions. There is a widespread belief that sensory symptoms characterize autism and differentiate it from other disorders. This paper examines the empirical evidence for this assumption. METHOD: All controlled experimental laboratory investigations published since 1960 were identified through systematic searches using Medline/PubMed and PsycInfo search engines. A total of 48 empirical papers and 27 theoretical or conceptual papers were reviewed. RESULTS: Sensory symptoms are more frequent and prominent in children with autism than in typically developing children, but there is not good evidence that these symptoms differentiate autism from other developmental disorders. Certain groups, including children with fragile X syndrome and those who are deaf blind, appear to demonstrate higher rates of sensory symptoms than children with autism. In reviewing the evidence relevant to two theories of sensory dysfunction in autism, over- and under-arousal theory, we find that there is very little support for hyper-arousal and failure of habituation in autism. There is more evidence that children with autism, as a group, are hypo-responsive to sensory stimuli, but there are also multiple failures to replicate findings and studies that demonstrate lack of group differences. CONCLUSIONS: The use of different methods, the study of different sensory modalities, and the changing scientific standards across decades complicate interpretation of this body of work. We close with suggestions for future research in this area. PMID- 16313427 TI - Parental imprisonment: effects on boys' antisocial behaviour and delinquency through the life-course. AB - BACKGROUND: Prisoners' children appear to suffer profound psychosocial difficulties during their parents' imprisonment. However, no previous study has examined later-life outcomes for prisoners' children compared to children separated from parents for other reasons. We hypothesise that parental imprisonment predicts boys' antisocial and delinquent behaviour partly because of the trauma of separation, partly because parental imprisonment is a marker for parental criminality, and partly because of childhood risks associated with parental imprisonment. METHOD: This study uses prospective longitudinal data from the Cambridge Study in Delinquent Development (CSDD). The CSDD includes data on 411 Inner London males and their parents. We compare boys separated by parental imprisonment during their first 10 years of life with four control groups: boys who did not experience separation, boys separated by hospital or death, boys separated for other reasons (usually disharmony), and boys whose parents were only imprisoned before their birth. Individual, parenting, and family risk factors for delinquency were measured when boys were aged 8-11. Eleven antisocial and delinquent outcomes were assessed between ages 14 and 40. RESULTS: Separation because of parental imprisonment predicted all antisocial-delinquent outcomes compared to the four control conditions. Separation caused by parental imprisonment was also strongly associated with many other childhood risk factors for delinquency. After controlling for parental convictions and other childhood risk factors, separation caused by parental imprisonment still predicted several antisocial-delinquent outcomes, even up to age 32, compared with other types of separation. CONCLUSIONS: Prisoners' children are a highly vulnerable group with multiple risk factors for adverse outcomes. Parental imprisonment appears to affect children over and above separation experiences and associated risks. Further research on possible moderating and mediating factors such as stigma, reduction in family income and reduced quality of care is required to identify the mechanisms by which parental imprisonment affects children. PMID- 16313428 TI - The Goteborg discotheque fire: posttraumatic stress, and school adjustment as reported by the primary victims 18 months later. AB - BACKGROUND: On October 29, 1998, around 400 young people were gathered in an old warehouse in Goteborg, Sweden, for a discotheque party. A fire erupted and spread explosively. Adolescents were exposed to dreadful scenes inside and outside the building. In all, 63 young people were killed and 213 physically injured. An 18 month follow-up with 275 adolescents (126 girls) who survived the fire, regarding the effects of the fire on symptoms of posttraumatic stress, school adjustment and performance, is reported. METHODS: Impact of Events Scale (IES), Clinician Administered Posttraumatic Stress Scale (CAPS) and an interview concerning background factors and issues of public and personal support. RESULTS: The level of posttraumatic stress was generally high, and highest among adolescents with an immigrant background. In all, 25% of the participants met DSM-IV criteria for PTSD. Twenty-three percent of the participants reported having either dropped out of school or repeated a class because of the fire. Students' ratings of how their own school handled the situation, and school absenteeism, were related to ratings of their own performance in school as well as to the level of posttraumatic stress. CONCLUSIONS: Traditional talking cures were more sought out by girls than boys. Broad-scale interventions must be complemented with special treatment opportunities for the most severely afflicted. Victims who suffer from high levels of posttraumatic stress need special support for a long period of time to minimise the effects on scholastic achievement and adjustment. Studies of the effects of traumatic events on child and adolescent development should measure school-related effects better than has hitherto been the case. PMID- 16313429 TI - Longitudinal improvements in communication and socialization of deaf children with cochlear implants and hearing aids: evidence from parental reports. AB - BACKGROUND: Research has shown that the cochlear implant may improve deaf children's speech and communication skills. However, little is known about its effect on children's ability to socialize with hearing peers. METHODS: Using a standardized psychological measure completed by parents and a longitudinal design, this study examined the development of communication, socialization, and daily living skills of children who used hearing aids or cochlear implants for an average of 11 and 6 years, respectively. RESULTS: Results show that children with cochlear implants, who were more delayed than children with hearing aids at the outset, made significant progress over time. Children with both devices achieved age-appropriate development after years of hearing aid or cochlear implant use. CONCLUSIONS: The pattern of results suggests that cochlear implants may be effective in improving deaf children's communication and social skills. PMID- 16313430 TI - Phoneme manipulation not onset-rime manipulation ability is a unique predictor of early reading. AB - BACKGROUND: Phonological awareness is known to be an excellent predictor of later reading acquisition. It remains unclear, however, whether phoneme manipulation alone best explains this association or whether an additional direct contribution of onset-rime awareness is predictive. This issue is explored here. METHOD: A longitudinal study is reported predicting national test and teacher-assessed performance of 351 children aged 7 from phonological awareness measures, pupil baseline attainment and background measures at age 5. RESULTS: Explicit phoneme manipulation skills at age 5 correlated most strongly with literacy skills at age 5. Phoneme manipulation at age 5 predicted all four reading measures taken at age 7 after pupil background, baseline data and onset-rime awareness were controlled in regression analyses. Onset-rime manipulation did not predict reading at 7 in parallel analyses. After controlling for initial reading at age 5, phoneme manipulation still predicted reading comprehension, and teacher-assessed reading and writing at age 7. CONCLUSIONS: Results support the existence of a route from phoneme manipulation, but not an additional direct route from explicit onset-rime manipulation at 5, to reading at 7. Practically, findings show that professionals can augment baseline and pupil background data with phoneme manipulation screening in the early identification of learning needs. PMID- 16313431 TI - Defining oppositional defiant disorder. AB - BACKGROUND: ICD-10 and DSM-IV include similar criterial symptom lists for conduct disorder (CD) and oppositional defiant disorder (ODD), but while DSM-IV treats each list separately, ICD-10 considers them jointly. One consequence is that ICD 10 identifies a group of children with ODD subtype who do not receive a diagnosis under DSM-IV. METHODS: We examined the characteristics of this group of children using the Great Smoky Mountains Study of children in the community aged 9-16. This study provided child and parent reports of symptoms and psychosocial impairment assessed with standardised diagnostic interviews. RESULTS: Children who received an ICD-10 diagnosis but not a DSM-IV diagnosis showed broadly similar levels of psychiatric comorbidity, delinquent activity and psychosocial impairment to those who met DSM-IV criteria in both cross-sectional and longitudinal analyses. CONCLUSIONS: These results indicate that DSM-IV excludes from diagnosis children who receive an ICD-10 diagnosis of CD (ODD sub-type), and who are substantially disturbed. Methods of redressing this situation are considered. PMID- 16313432 TI - The developmental association between eating disorders symptoms and symptoms of depression and anxiety in juvenile twin girls. AB - OBJECTIVE: We investigated the role of genetic and environmental factors in the developmental association among symptoms of eating disorders, depression, and anxiety syndromes in 8-13-year-old and 14-17-year-old twin girls. METHODS: Multivariate genetic models were fitted to child-reported longitudinal symptom data gathered from clinical interview on 408 MZ and 198 DZ female twin pairs from the Virginia Twin Study of Adolescent Behavioural Development (VTSABD). RESULTS: Model-fitting revealed distinct etiological patterns underlying the association among symptoms of eating disorders, depression, overanxious disorder (OAD), and separation anxiety disorder (SAD) during the course of development: 1) a common genetic factor influencing liability to all symptoms - of early and later OAD, depression, SAD, and eating symptoms; 2) a distinct genetic factor specifically indexing liability to early eating disorders symptoms; 3) a shared environmental factor specifically influencing early depression and early eating disorders symptoms; and 4) a common environmental factor affecting liability to symptoms of later eating disorders and both early and later separation anxiety. CONCLUSIONS: These results suggest a pervasive genetic effect that influences liability to symptoms of over-anxiety, separation anxiety, depression, and eating disorder throughout development, a shared environmental influence on later adolescent eating problems and persistent separation anxiety, genetic influences specific to early eating disorders symptoms, and a shared environmental factor influencing symptoms of early eating and depression. PMID- 16313433 TI - Tourette's syndrome: performance on tests of behavioural inhibition, working memory and gambling. AB - BACKGROUND: Tourette's syndrome (TS) is a neurodevelopmental disorder associated with fronto-striatal dysfunction. There is debate as to the extent to which TS is associated with cognitive impairment. Some authors argue that any impairments seen are attributable to comorbid psychiatric symptomatology, whilst others have suggested that uncomplicated TS is associated with mild deficits limited primarily to inhibitory processes. The present study was designed to examine this issue using carefully screened participants with TS and experimental measures thought to involve different areas of the frontal lobes. METHODS: Adolescents with TS who were screened for comorbid psychiatric disorder were compared with a healthy control group on a set of executive measures. Two tasks involving behavioural inhibition were used: a Sentence Completion task in which sentences had to be completed first with straightforward and then with nonsensical endings, and a Flanker task in which a central stimulus was surrounded by either compatible or incompatible flankers. Working memory was assessed using an N-back task, and reward learning was assessed using a Gambling task. Both accuracy and reaction times were measured for each task. RESULTS: The TS group differed significantly from the control group on both the Sentence Completion task and on the Flanker task. On the Sentence Completion task, they were slower to make both sensible and nonsensical completions, and they had higher error scores on the nonsensical completions. On the Flanker task, the TS participants were less accurate than the control group, since they were poorer on the incompatible but not the compatible trials. A similar interaction with trial type was found for speed, where the TS participants were slowed more by the incompatible versus compatible trials, although overall their performance tended to be faster than the controls. The TS group did not differ significantly from the control group on measures of working memory or reward learning. CONCLUSIONS: The findings provide further evidence that uncomplicated TS is not associated with widespread executive impairments. However, it was not clear that any differences between the groups could be attributed solely to selective inhibitory impairment. PMID- 16313434 TI - Examining the relationship between rightward visuo-spatial bias and poor attention within the normal child population using a brief screening task. AB - BACKGROUND: Some previous studies have linked Attention Deficit Hyperactivity Disorder (ADHD) with a bias in spatial awareness away from the left. As genetic research suggests that ADHD may be better viewed as an extreme on a continuum rather than a distinct entity, here we examined this issue in boys from the normal population. METHOD: From an initial sample of 1811, two groups of boys characterised by very high (n = 58) or very low (n = 68) levels of ADHD-type behaviours were formed. The groups completed the spatially sensitive Line Bisection test and more general measures of (non-spatial) attention and intellectual function. RESULTS: Boys whose bisections were consistent with relative inattention to the left indeed had higher ratings of ADHD-type behaviours and performed significantly more poorly on tests of sustained attention and executive function than boys whose bisections were in the normal range. In contrast, boys who showed extreme bisections in the opposite direction were not unusual either in ratings or test performance. CONCLUSIONS: The results support an association between poor attention and a relative rightward bias in visual awareness that may stem from right hemisphere inefficiency. PMID- 16313435 TI - Insights into the control of attentional set in ADHD using the attentional blink paradigm. AB - BACKGROUND: Previous work on visual selective attention in Attention Deficit Hyperactivity Disorder (ADHD) has utilised spatial search paradigms. This study compared ADHD to control children on a temporal search task using Rapid Serial Visual Presentation (RSVP). In addition, the effects of irrelevant singleton distractors on search performance were examined. METHOD: In each condition, subjects reported the identity of a red letter 'probe' in a sequence of white letters which appeared one after the other at a central fixation point. The temporal position of the probe varied from an initial target, which was distinguished by surrounding asterisks. The target was reported in addition to the probe in condition 2, but not in the baseline condition 1. In a third condition, the initial target was not reported, but one of the asterisks appeared as a colour singleton on some trials. RESULTS: All children displayed an 'attentional blink' with probe detection reduced when it appeared at close temporal relations relative to the target. This 'blink' reduced over time, and there were no group differences in the recovery of performance, although ADHD children made more errors overall. The ADHD group were also more vulnerable than controls to distraction from irrelevant singletons in condition 3. CONCLUSION: Although the basic mechanisms of selective attention were not impaired in children with ADHD, these children appeared to require more resources to execute the task and were more vulnerable to distraction by irrelevant singletons, indicating deficits in the maintenance of attentional control. PMID- 16313436 TI - Saccadic eye movement task identifies cognitive deficits in children with schizophrenia, but not in unaffected child relatives. AB - BACKGROUND: The delayed oculomotor response (DOR) task requires response inhibition followed by movement of gaze towards a known spatial location without a current stimulus. Abnormalities in response inhibition and in the spatial accuracy of the eye movement are found in individuals with schizophrenia and in many of their relatives, supporting the use of these saccadic abnormalities as endophenotypes in genetic studies. It is unknown whether school-age children, either with psychosis or as relatives of a schizophrenic proband, can be included. METHOD: One hundred eighty-seven children, ages 5.8-16.0 years - 45 children with childhood-onset schizophrenia, 64 children with a first-degree relative with schizophrenia, and 84 typically developing children - completed DOR tasks with 1 and 3 second delays. RESULTS: Children with childhood-onset schizophrenia demonstrated impaired response inhibition and impaired spatial accuracy compared to both relatives and typicals; however, relatives and typicals did not differ from each other. CONCLUSIONS: Children with childhood-onset schizophrenia have saccadic abnormalities similar to those found in adults with schizophrenia, supporting the continuity of executive function deficits in childhood-onset with adolescent and adult-onset schizophrenia. However, saccadic tasks are not sensitive to genetic risk in non-psychotic children and 6-15-year old children should not be included in genetic studies utilizing this endophenotype. PMID- 16313437 TI - Actin-based motility in the net slime mould Labyrinthula: evidence for the role of myosin in gliding movement. AB - In contrast to crawling movement (e.g. in amoebae and tissue cells) the other major class of substratum-associated motility in eukaryotes, gliding, has received relatively little attention. The net slime mold Labyrinthula provides a useful laboratory model for studying this process since it exhibits a particular kind of gliding in its plasmodial stage. Here nucleated spindle cells glide along self-established cytoplasmic trackways in a predominantly unidirectional manner, at 1-2 microm/s. These trackways, upon which gliding is dependent, are held by filopodial tethers some distance off the well-developed reticulopodial mesh anchoring the plasmodium onto the substratum. Reflection interference microscopy resolves this matrix in live plasmodia. The axially disposed cytoskeletal elements of the trackways are revealed by rhodamine-labelled phalloidin to be rich in F-actin. A weft of peripheral, rapidly extending filopodia (50 microm/min) typifies the expanding regions of the plasmodium. Here spindle cells are recruited before emigrating into newly differentiated trackways. Immunoblotting whole plasmodia or a sucrose-soluble cytoplasmic extract reveals a single actin-positive band of Mr 48 kDa. Polyclonal antibodies to two distinct myosin peptide sequences identify a single myosin HC (Mr 96 kDa) in immunoblots. Gliding was reversibly blocked by 10 mM 2,3-butanedione-2-monoxime, a myosin ATPase inhibitor, but it was insensitive to the actin-binding drugs cytochalasin D and phalloidin. We suggest that the force (>50 pN) for gliding motility results from interaction of myosin molecules, associated with the spindle cells, with trackway F-actin via the bothrosomes. PMID- 16313438 TI - Myosporidium merluccius n. g., n. sp. infecting muscle of commercial hake (Merluccius sp.) from fisheries near Namibia. AB - A new species of Microsporidia classified to a new genus was observed in the trunk muscle of commercial hake (Merluccius capensis/paradoxus complex) from Namibian fisheries. Macroscopic examination revealed thin and dark filaments inserted among muscle fibers. Inside the filaments were many sporophorous vesicles with about 30-50 spores per vesicle. The shape of the spore was pyriform and the extruded polar filament was of moderate length (up to 4.29 microm, n=12). This new species of Microsporidia is described using macrophotography, microphotography, staining, and transmission electron microscopy (TEM), as well as molecular methods. Its 16S rRNA was found to be similar to that of Microsporidium prosopium Kent et al., 1999, while both sequences were quite different from 16S rRNA sequences known for other Microsporidia. Nevertheless, this new species is separated morphologically from M. prosopium by the presence of 11-12 anisofilar coils and the formation of the xenoma at the site of infection. Type species. PMID- 16313439 TI - The effects of E-beam irradiation and microwave energy on Eastern Oysters (Crassostrea virginica) experimentally infected with Cryptosporidium parvum. AB - Shellfish have been identified as a potential source of Cryptosporidium infection for humans. The inactivation of C. parvum and other pathogens in raw molluscan shellfish would provide increased food safety for normal and at-risk consumers. The present study examined the efficacy of two alternative food-processing treatments, e-beam irradiation and microwave energy, on the viability of C. parvum oocysts in Eastern Oysters (Crassostrea virginica), which were artificially infected with the Beltsville strain of C. parvum. The effects of the treatments were evaluated by oral feeding of the processed oyster tissues to neonatal mice. Significant reductions (P<0.05) in infectivity were observed for in-shell and shucked oysters treated with e-beam irradiation at doses of 1.0, 1.5, or 2 kGy vs. untreated controls. A dose of 2 kGy completely eliminated C. parvum infectivity and did not adversely affect the visual appearance of the oysters. Oyster tissue treated with microwave exposures of 1 s (43.2 degrees C), 2 s (54.0 degrees C), and 3 s (62.5 degrees C) showed a reduction in C. parvum mouse infectivity, but the effects were not significantly different (P>0.05) from controls. Microwave energy treatments at 2 and 3 s showed extensive changes in oyster meat texture and color. Thus, because of lack of efficacy and unacceptable tissue changes, microwave treatment of oysters is not considered a viable food processing method. PMID- 16313440 TI - In vitro and ex vivo permissivity of hepatocytes for Leishmania donovani. AB - Using models of ex vivo infection of murine, rat, and human primary hepatocytes by Leishmania donovani, we showed that hepatocytes are permissive for Leishmania at a low level. We then modeled the in vitro infection of a human hepatoma derived cell line to examine the parasite's capability to proliferate and to cause direct damage to hepatocytes. Results showed that L. donovani can infect hepatocytes, but do not massively proliferate. This slight infection under our experimental conditions resulted in limited damage to hepatocytes. These results bring into question a possible role for hepatocytes as a parasite reservoir during latent infection. PMID- 16313441 TI - Assessment of the cell viability of cultured Perkinsus marinus (Perkinsea), a parasitic protozoan of the Eastern oyster, Crassostrea virginica, using SYBRgreen propidium iodide double staining and flow cytometry. AB - A flow cytometry (FCM) assay using SYBRgreen and propidium iodide double staining was tested to assess viability and morphological parameters of Perkinsus marinus under different cold- and heat-shock treatments and at different growth phases. P. marinus meront cells, cultivated at 28 degrees C, were incubated in triplicate for 30 min at -80 degrees C, -20 degrees C, 5 degrees C, and 20 degrees C for cold-shock treatments and at 32 degrees C, 36 degrees C, 40 degrees C, 44 degrees C, 48 degrees C, 52 degrees C, and 60 degrees C for heat-shock treatments. A slight and significant decrease in percentage of viable cells (PVC), from 93.6% to 92.7%, was observed at -20 degrees C and the lowest PVC was obtained at -80 degrees C (54.0%). After 30 min of heat shocks at 40 degrees C and 44 degrees C, PVC decreased slightly but significantly compared to cells maintained at 28 degrees C. When cells were heat shocked at 48 degrees C, 52 degrees C, and 60 degrees C heavy mortality occurred and PVC decreased to 33.8%, 8.0%, and 3.4%, respectively. No change in cell complexity and size was noted until cells were heat shocked at >or=44 degrees C. High cell mortality was detected at stationary phase of P. marinus cell culture. Cell viability dropped below 40% in 28-day-old cultures and ranged 11-25% in 38 to 47-day-old cultures. Results suggest that FCM could be a useful tool for determining viability of cultured P. marinus cells. PMID- 16313442 TI - The effect of high-pressure processing on infectivity of Cryptosporidium parvum oocysts recovered from experimentally exposed Eastern oysters (Crassostrea virginica). AB - Shellfish have been identified as a potential source of Cryptosporidium infection for humans. The inactivation of Cryptosporidium parvum and other pathogens in raw molluscan shellfish would provide increased food safety for normal and at-risk consumers. The present study identified the efficacy of a non-thermal alternative food-processing treatment, high hydrostatic pressure processing (HPP), on the viability of C. parvum oocysts in the Eastern oysters Crassostrea virginica. Oysters were artificially exposed to 2 x 10(7) oocysts of the Beltsville strain of C. parvum in seawater and subjected to HPP treatments. The effects of the treatments were evaluated by inoculation of the processed oyster tissues into neonatal mice. High-pressure processing of shucked Eastern oysters at all pressures tested (305, 370, 400, 480, and 550 MPa) was significantly effective (P<0.05) in reducing the numbers of positive mouse pups fed treated oyster tissues exposed to C. parvum oocysts. A dose of 550 MPa at 180 s (s) of holding time produced the maximum decrease in numbers of C. parvum positive mouse pups (93.3%). Measurement of tristimulus color values of HPP-treated raw oysters at extended processing times from 120 s to 360 s at 550 MPa showed a small increase in whiteness of oyster meat. This non-thermal processing treatment shows promise for commercial applications to improve safety of seafood and reduce public health risks from cryptosporidiosis. PMID- 16313443 TI - Complete rRNA gene sequences reveal that the microsporidium Nosema bombi infects diverse bumblebee (Bombus spp.) hosts and contains multiple polymorphic sites. AB - Characterisation of microsporidian species and differentiation among genetic variants of the same species has typically relied on ribosomal RNA (rRNA) gene sequences. We characterised the entire rRNA gene of a microsporidium from 11 isolates representing eight different European bumblebee (Bombus) species. We demonstrate that the microsporidium Nosema bombi infected all hosts that originated from a wide geographic area. A total of 16 variable sites (all single nucleotid polymorphisms (SNPs)) was detected in the small subunit (SSU) rRNA gene and 42 (39 SNPs and 3 indels) in the large subunit (LSU) rRNA sequence. Direct sequencing of PCR-amplified DNA products of the internal transcribed spacer (ITS) region revealed identical sequences in all isolates. In contrast, ITS fragment length determined by PAGE and sequencing of cloned amplicons gave better resolution of sequences and revealed multiple SNPs across isolates and two fragment sizes in each isolate (six short and seven long amplicon variants). Genetic variants were not unique to individual host species. Moreover, two or more sequence variants were obtained from individual bumblebee hosts, suggesting the existence of multiple, variable copies of rRNA in the same microsporidium, and contrary to that expected for a class of multi-gene family under concerted evolution theory. Our data on within-genome rRNA variability call into question the usefulness of rRNA sequences to characterise intraspecific genetic variants in the Microsporidia and other groups of unicellular organisms. PMID- 16313444 TI - Parastrombidinopsis shimi n. gen., n. sp. (Ciliophora: Choreotrichia) from the coastal waters of Korea: morphology and small subunit ribosomal DNA sequence. AB - The planktonic ciliate Parastrombidinopsis shimi n. gen., n. sp. is described from both living cells and quantitative protargol-stained (QPS) preparations and the sequence of the small subunit rDNA (SSU rDNA) is reported. This species is almost oval when the cells are alive; when stained, it is cylindrical for the upper two-fifths, half-bowl shaped for the middle two-fifths, and narrow rodshaped for the lower one-fifth. The ranges (and mean +/- standard deviation, n = 20) of cell length, cell width, and oral diameter of living cells were 112-221 microm (168 +/- 39), 88-176 microm (121 +/- 30), and 53-110 microm (80 +/- 14), respectively, while those of the QPS-stained specimens (n = 54) were 88-225 microm (162 +/- 29), 55-163 microm (102 +/- 19), and 53-98 microm (69 +/- 9), respectively. Thirty-six to 48 external oral polykinetids had cilia 25-40 microm long. However, unlike Strombidinopsis species sensu stricto, P. shimi has an external oral polykinetid zone that is an open circle. This species has two shorter polykinetids associated with the end of the oral polykinetid zone, deep in the oral cavity. Like Strombidinopsis species in the subclass Choreotrichia, 36-50 somatic kineties were equally spaced around the cell body and extended from the oral to the posterior regions with 68-105 dikinetids per kinety. Both kinetosomes of each kinetid bore cilia 3-10 microm long. Parastrombidinopsis shimi had 2 (1-4) ovoid macronuclei of 20-82 x 15-32 microm. When properly aligned, the sequence of the SSU rDNA of P. shimi (GenBank Accession No. AJ786648) was approximately 5% different from that of Strobilidium caudatum and 6% different from that of two Strombidinopsis species. Based both on morphology and gene sequence divergence, we establish this is as a new species in a new genus belonging to the family Strombidinopsidae. PMID- 16313445 TI - Morphologic and molecular identification of Naegleria dunnebackei n. sp. isolated from a water sample. AB - Naegleria dunnebackei n. sp., a new species of the free-living amoeboflagellate Naegleria, is described in this report. The organism was isolated from a water sample taken from drinking troughs associated with cases of primary amoebic meningoencephalitis in cattle at a ranch in southern California. The isolate grew at, but not above 37 degrees C, and did not kill young mice upon intranasal inoculation suggesting that it was not pathogenic. The new species combines morphological features of non-pathogenic Naegleria gruberi and pathogenic Naegleria fowleri. The trophic amoeba resembled other members of the genus, with a prominent vesicular nucleus and mitochondria with discoidal cristae; a Golgi apparatus was not observed by electron microscopy. The cyst stage had pores in the wall typical of those seen in pathogenic N. fowleri. Upon suspension in distilled water, amoebae transformed into temporary, non-feeding flagellates, mostly with two anterior flagella but occasionally with four. The rationale for its description as a new species was based upon sequencing of the 5.8S rDNA and internal transcribed spacers of the amoeba, which is similar to but not identical to that of Naegleria gallica, differing from that organism's DNA by six base pairs. Virus-like elements were found in the cytoplasm of trophic amoebae, often in association with crystalloids, and may be the cause of lysis of amoebae in culture. PMID- 16313446 TI - Ultrastructural studies of Henneguya rhamdia n. sp. (Myxozoa) a parasite from the Amazon teleost fish, Rhamdia quelen (Pimelodidae). AB - Henneguya rhamdia n. sp. is described in the gill filaments of the teleost fish Rhamdia quelen, collected from the Peixe Boi River, State of Para, Brazil. This myxosporean produced spherical to ellipsoidal plasmodia, up to 300 microm in diameter, which contained developmental stages, including spores. Several dense bodies up to 2 microm in diameter were observed among the spores. The spore body was ellipsoidal (13.1 microm in length, 5.2 microm in width, and 2.5 microm in thickness) and each of the two valves presented a tapering tail (36.9 microm in length). These valves surrounded the binucleated sporoplasm cell and two equal ellipsoidal polar capsules (4.7 x 1.1 microm), which contained 10-11 (rarely 12) polar filament coils. The sporoplasm contained sporoplasmosomes with a laterally eccentric dense structure with a half-crescent section. Based on the data obtained by electron microscopy and on the host specificity, the spores differed from previously described Henneguya species, mainly in their shape and size, number and arrangement of the polar filament coils, and sporoplasmosome morphology. PMID- 16313447 TI - Mitochondrial cytochrome b mRNA editing in dinoflagellates: possible ecological and evolutionary associations? AB - To verify the hypothesis that mt mRNA editing is widespread in dinoflagellates, we analyzed cytochrome b (cob) mRNA editing for six species representing distinct ecotypes and taxonomic classes of Dinophyceae. Editing is detected in all, which is similar to the three other species studied previously in that edited sites appear to aggregate in four clusters and occur predominantly at first and second positions of codons (93%), overwhelmingly involving A --> G, U --> C, or C --> U substitutions with a smaller number of G --> C, G --> A changes. Comparative analyses on editing characteristics reveal interesting trends related to phylogenetic relatedness and ecological features. Editing density (percentage of nucleotide that is affected by editing) increases from early to derived lineages. Higher editing densities also map to red tide-forming lineages. Furthermore, similarity of location of edited codons (LOE) and the type of nucleotide changes (TOE) in different lineages mirror the taxonomic affinity of the lineages. Phylogenetic trees constructed from LOE and TOE resemble those inferred from cob sequences. The results bolster our earlier hypothesis that cob editing is widespread in dinoflagellates and suggest that density, location, and type of editing may bear yet-to-be-defined evolutionary and ecological significance. PMID- 16313450 TI - Adaptive radiation in microbial microcosms. AB - It has often been argued that evolutionary diversification is the result of divergent natural selection for specialization on alternative resources. I provide a comprehensive review of experiments that examine the ecology and genetics of resource specialization and adaptive radiation in microbial microcosms. In these experiments, resource heterogeneity generates divergent selection for specialization on alternative resources. At a molecular level, the evolution of specialization is generally attributable to mutations that de regulate the expression of existing biosynthetic and catabolic pathways. Trade offs are associated with the evolution of resource specialization, but these trade-offs are often not the result of antagonistic pleiotropy. Replicate adaptive radiations result in the evolution of a similar assemblage of specialists, but the genetic basis of specialization differs in replicate radiations. The implications of microbial selection experiments for evolutionary theory are discussed and future directions of research are proposed. PMID- 16313451 TI - Molecular evolution of animal antimicrobial peptides: widespread moderate positive selection. AB - An increasing number of studies in both vertebrates and invertebrates show that the evolution of antimicrobial peptides is driven by positive selection. Because these diverse molecules show potential for therapeutic applications, they are currently the targets of much structural and functional research, providing extensive background data for evolutionary studies. In this paper, patterns of molecular evolution in antimicrobial peptide genes are reviewed. Evidence for positive selection on antimicrobial peptides includes an excess of nonsynonymous nucleotide substitutions, an excess of charge-changing amino acid substitutions, nonneutral patterns of allelic variation, and functional assays in vivo and in vitro that show improved antimicrobial effects for derived sequence variants. Positive selection on antimicrobial peptides may be as common as, but perhaps weaker than, selection on the best-known example of adaptively evolving immunity genes, the major histocompatibility complex. Thus, antimicrobial peptides present a useful and underutilized model for the study of adaptive molecular evolution. PMID- 16313452 TI - The effect of female polyandry and sperm precedence on the evolution of sexual difference in dispersal timing. AB - Predispersal copulation and unpredictable environment facilitate the evolution of female-biased dispersal in species, where females are functionally monandrous. Females should migrate and reproduce over different habitats to spread their risks due to environmental fluctuation. On the other hand, males do not have to disperse because their risks are spread by their mating partners who produce their offspring in different habitats. However, when females are functionally polyandrous, it is expected that they will not contribute to spreading the male's risk extensively. Therefore, by simulation with the individual based model, the present study evaluated how female polyandry influences the sexual difference in dispersal timing. This model revealed that when females are polyandrous, the timing of female remating and sperm priority patterns have an important influence on the evolution of sex-biased dispersal. Particularly when female remating is not synchronized with dispersal or when last-male sperm precedence does not exist, female-biased dispersal is evolved. PMID- 16313453 TI - Indirect consequences of artificial selection on plasticity to light quality in Arabidopsis thaliana. AB - Covariation between light quality- and photoperiod-mediated phenotypic plasticity was investigated using Arabidopsis thaliana. Three episodes of artificial selection were imposed on an index that quantified the plastic response to reduced red to far-red ratios (R:FR), with higher index values indicating greater plasticity. Relative to control lines, two high plasticity (HP) lines showed 1.6- and 2.4-fold increases in the index; low plasticity (LP) lines showed 1.4- and 1.1-fold decreases. A factorial experiment combining high and low R:FR conditions with long and short photoperiods assessed indirect consequences of selection on plasticity. Despite divergent R:FR-mediated plasticities in HP vs. LP lines, all four lines showed increases in photoperiod-mediated responses and decreases in mean leaf number. Complex relationships among trait means, plasticities and underlying mechanisms caution against generalizing about the genetic architecture of plastic traits. Partially independent developmental and evolutionary responses to R:FR and photoperiod are somewhat unsurprising, given this species' cosmopolitan nature. PMID- 16313454 TI - Amount of introgression in flycatcher hybrid zones reflects regional differences in pre and post-zygotic barriers to gene exchange. AB - Introgression is the incorporation of alleles from one species or semispecies into the gene pool of another through hybridization and backcrossing. The rate at which this occurs depends on the frequency of hybridization and the fitness of hybrids and backcrosses compared to 'pure' individuals. The collared flycatcher (Ficedula albicollis) and the pied flycatcher (F. hypoleuca) co-exist and hybridize at low to moderate frequencies in a clinal hybrid zone in Central Europe and on the islands of Gotland and Oland off the Swedish east coast. Data on hatching success suggest that hybrids are less fertile in Central Europe compared to on the islands. Direct fitness estimates using molecular markers to infer paternity are consistent with the demographic data. Applying a tag-array based minisequencing assay to genotype interspecific substitutions and single nucleotide polymorphisms we demonstrate that the amount of introgression from the pied to the collared flycatcher is higher in the two island populations (Gotland and Oland) than in two geographically distinct areas from the Central European hybrid zone (Czech Republic and Hungary). In all areas the amount of introgression from collared to pied flycatchers is very low or seemingly absent. The different patterns of introgression are consistent with regional differences in rates of hybridization and fitness of hybrids. We suggest that barriers to gene exchange may have been partly broken down on the islands due to asymmetric gene flow from allopatry. Alternatively, or in addition, more pronounced reinforcement of prezygotic isolation in Central Europe might have increased post zygotic isolation through hitchhiking, since genes affecting pre and post-zygotic isolation are both sex-linked in these birds. One of our genetic markers appears to introgress from pied to collared flycatchers at a much higher rate than the other markers. We discuss the possibility that the introgressed marker may be linked to a gene which is under positive selection in the novel genetic background. PMID- 16313455 TI - The evolution of fledging age in songbirds. AB - In birds with altricial young an important stage in the life history is the age at fledging. In this paper we use an approach proven successful in the prediction of the optimal age at maturity in fish and reptiles to predict the optimal age of fledging in passerines. Integrating the effects of growth on future fecundity and survival leads to the prediction that the optimal age at fledging is given by a function that comprises survival to maturity, the exponent of the fecundity-body size relationship and nestling growth. Growth is described by the logistic equation with parameters, A, K and t(i). Assuming that the transitional mortality curve can be approximated by the nestling mortality, M(n), the optimal fledging age, t(f), is given by a simple formula involving the three growth parameters, nestling mortality (M(n)) and the exponent (d) of the fecundity-body size relationship. Predictions of this equation underestimate the true values by 11 16%, which is expected as a consequence of the transitional mortality function approximation. A transitional mortality function in which mortality is approximately 0.3-0.4 of nesting mortality (i.e. mortality declines rapidly after fledging) produces predictions which, on average, equal the observed values. Data are presented showing that mortality does indeed decline rapidly upon fledging. PMID- 16313456 TI - Inbreeding depression and heterosis in populations of Schiedea viscosa, a highly selfing species. AB - Progeny from self-pollinations and intrapopulation crosses were examined in Schiedea viscosa to determine the extent of inbreeding depression in this highly selfing species. Progeny of interpopulation crosses were also investigated to determine whether deleterious alleles have been fixed in populations of S. viscosa. There was no inbreeding depression at early life history stages, including seeds per capsule, seed mass, or germination. Inbreeding depression was detected for the later life history stage of fruit biomass, although not for survival or life span. Heterosis for vegetative biomass and fruit biomass was detected in progeny from crosses between populations. Levels of inbreeding depression in S. viscosa are low relative to out-crossing species of Schiedea, especially when early life history stages are compared. PMID- 16313457 TI - Local adaptation and population structure at a micro-geographical scale of a fungal parasite on its host plant. AB - Local adaptation, which has been detected for several wild pathosystems is influenced by gene flow and recombination. In this study, we investigate local adaptation and population structure at a fine scale in wild populations of a plant-pathogen fungus. We sampled hierarchically strains of Colletotrichum lindemuthianum in a wild population of its host. The analysis of AFLP patterns obtained for 86 strains indicated that: (i) many different haplotypes can be discriminated, although occurrence of recombination could not be shown; (ii) migration between adjacent plants seemed rare during the season; and (iii) neutral diversity is structured according to groups of plants and individual host plants. Furthermore, we tested for the occurrence of local adaptation using a cross-inoculation experiment. Our results showed local adaptation at the scale of the individual host plant. These results indicate that fine-scale dynamics has evolutionary consequences in this pathosystem. PMID- 16313458 TI - Female common lizards (Lacerta vivipara) do not adjust their sex-biased investment in relation to the adult sex ratio. AB - Sex allocation theory predicts that facultative maternal investment in the rare sex should be favoured by natural selection when breeders experience predictable variation in adult sex ratios (ASRs). We found significant spatial and predictable interannual changes in local ASRs within a natural population of the common lizard where the mean ASR is female-biased, thus validating the key assumptions of adaptive sex ratio models. We tested for facultative maternal investment in the rare sex during and after an experimental perturbation of the ASR by creating populations with female-biased or male-biased ASR. Mothers did not adjust their clutch sex ratio during or after the ASR perturbation, but produced sons with a higher body condition in male-biased populations. However, this differential sex allocation did not result in growth or survival differences in offspring. Our results thus contradict the predictions of adaptive models and challenge the idea that facultative investment in the rare sex might be a mechanism regulating the population sex ratio. PMID- 16313459 TI - Phenotypic and genetic variation in emergence and development time of a trimorphic damselfly. AB - Although colour polymorphisms in adult organisms of many taxa are often adaptive in the context of sexual selection or predation, genetic correlations between colour and other phenotypic traits expressed early in ontogeny could also play an important role in polymorphic systems. We studied phenotypic and genetic variation in development time among female colour morphs in the polymorphic damselfly Ischnura elegans in the field and by raising larvae in a common laboratory environment. In the field, the three different female morphs emerged at different times. Among laboratory-raised families, we found evidence of a significant correlation between maternal morph and larval development time in both sexes. This suggests that the phenotypic correlation between morph and emergence time in the field has a parallel in a genetic correlation between maternal colour and offspring development time. Maternal colour morph frequencies could thus potentially change as correlated responses to selection on larval emergence dates. The similar genetic correlation in male offspring suggests that sex-limitation in this system is incomplete, which may lead to an ontogenetic sexual conflict between selection for early male emergence (protandry) and emergence times associated with maternal morph. PMID- 16313460 TI - On the worldwide spread of an insecticide resistance gene: a role for local selection. AB - Adaptation occurs by gene replacement (or transient balanced polymorphism). Replacement may be caused by selection (local or global) and/or genetic drift among alleles. In addition, historical events may blur the respective effects of selection and drift during the course of replacement. We address the relative importance of these processes in the evolution of insecticide resistance genes in the mosquito Culex pipiens. The resistance allele, Ester2, has a broad geographic distribution compared to the other resistance alleles. To distinguish between the different processes explaining this distribution, we reviewed the literature and analysed updated data from the Montpellier area of southern France. Overall, our data indicate that Ester2 prevails over other Ester resistance alleles in moderately treated areas. Such conditions are common and favour the hypothesis of selection acting at a local level. This places an emphasis on the importance of ecological conditions during the evolution of resistance. Finally, we highlight that historical events have contributed to its spread in some areas. PMID- 16313461 TI - Asymmetrical reproductive character displacement in the house mouse. AB - Our study addressed reproductive character displacement between two subspecies of the house mouse, Mus musculus musculus and Mus musculus domesticus that hybridize in Europe along a zone where selection against hybridization is known to occur. Based on a multi-population approach, we investigated spatial patterns of divergence of mate preference in the two taxa. Mate preference was significantly higher in the contact zone than in allopatry in both subspecies, suggesting that reproductive character displacement occurs. Moreover, patterns of preference were stronger in M. m. musculus than in M. m. domesticus, indicating an asymmetrical divergence between the two. In the context of selection against hybridization, our results may provide empirical support for the hypothesis of reinforcement in a parapatric hybrid zone. We discuss factors that could explain the asymmetrical pattern of divergence and the possible impact of a unimodal structure on the maintenance of premating divergence between the two subspecies. PMID- 16313462 TI - Influence of genotype-temperature interaction on pollen performance. AB - Pollen competition and selection have significant evolutionary consequences, but very little is known about how they can be modulated. We have examined in cherry (Prunus avium L.) how pollen performance is affected by the genotype of the pollen and by the environmental conditions under which it grows, namely the pistilar tissue and temperature. The different pollen donor genotypes tested in this work differed in their behaviour both in vitro and in vivo and this behaviour was modulated depending on the female recipient they grew on. Furthermore, there was a significant temperature-genotype interaction that affected the pollen tube population census that succeeded in reaching the base of the style. The combination of these three factors, while enabling a capacity of response to variations in environmental pressures, could maintain variability in pollen performance avoiding the fixation of the genes that control pollen tube growth rate. PMID- 16313463 TI - Evolutionary dynamics and population biology of a polymorphic insect. AB - Conspicuous heritable polymorphisms are useful to address the question if morph frequencies are stable or whether they fluctuate between generations. Ecological geneticists have studied colour polymorphisms in the past, but there are few long term studies of genetic dynamics across multiple generations. We studied morph frequency dynamics and female fecundity in the trimorphic blue-tailed damselfly (Ischnura elegans). The morphs include a male-coloured (androchrome) type of female, which is thought to be maintained by frequency-dependent sexual conflict. Morph frequencies changed significantly between years across all populations. There was evidence for directional frequency change since androchrome females increased in 9 of 10 populations across a 4-year period. There was heterogeneity between populations in their evolutionary trajectories, partly caused by population age: androchrome frequencies were initially high in young populations but gradually decreased and approached the level of old populations. We discuss the possible causes of morph-frequency fluctuations, and the role of morph specific fecundity, dispersal and other forces influencing evolutionary dynamics in this system. PMID- 16313464 TI - Sexual conflict and female immune suppression in the cricket, Allonemobious socius. AB - In many animal systems, females exhibit a localized immune response to insemination that helps defend against sexually transmitted disease. However, this response may also kill sperm, reducing a male's reproductive potential. If males could suppress this response, they may be able to increase their sperm's representation in the female's reproductive tract, thereby increasing their fitness. Here we address the hypothesis that, under conditions of sperm competition, males interfere with female immunity. To test our hypothesis, we manipulated levels of female mating frequency (single vs. multiply mated) and seminal diversity (monandrous vs. polyandrous) in the cricket, Allonemobius socius and measured female immune response. As mating frequency increased, female hemocyte load decreased, indicating a general reproductive cost. As seminal diversity increased, phenoloxidase (PO) activity (in vitro measure of 'potential' macroparasitic defense) increased and encapsulation ability (in vivo measure of 'realized' macroparasitic defense) decreased in polyandrous females. These results suggest that males may manipulate female immunity by interrupting the pro PO cascade, which begins with the activation of PO and ends in the encapsulation of invading foreign bodies. In other words, female immune function may serve as a battleground over which a sexual conflict is fought. PMID- 16313465 TI - Games among cannibals: competition to cannibalize and parent-offspring conflict lead to increased sibling cannibalism. AB - Sibling cannibalism occurs in many species, yet understanding of sibling cannibalism as an adaptation currently lags behind understanding of other antagonistic interactions among siblings. Observed sibling cannibalism phenotypes likely reflect the interaction between competitive games among siblings and parent-offspring conflict. Using a game-theoretic approach, we derive optimal offspring cannibalism behaviour and parental modifiers that limit or facilitate cannibalism. The results are compared to contemporary frequency-independent analysis. With the addition of game interactions among siblings or parent offspring co-evolution, our model predicts increased cannibalism (compared to the frequency-independent prediction), as offspring compete to eat siblings. When infertile eggs are present--strengthening competition--offspring risk eating viable siblings in order to gain access to infertile eggs, intensifying parent offspring conflict. We use the results to make new predictions about the occurrence of sibling cannibalism. Additionally, we demonstrate the utility of trophic egg laying as a maternal mechanism to promote egg eating. PMID- 16313466 TI - Interspecific variation in the use of carotenoid-based coloration in birds: diet, life history and phylogeny. AB - Birds show striking interspecific variation in their use of carotenoid-based coloration. Theory predicts that the use of carotenoids for coloration is closely associated with the availability of carotenoids in the diet but, although this prediction has been supported in single-species studies and those using small numbers of closely related species, there have been no broad-scale quantitative tests of the link between carotenoid coloration and diet. Here we test for such a link using modern comparative methods, a database on 140 families of birds and two alternative avian phylogenies. We show that carotenoid pigmentation is more common in the bare parts (legs, bill and skin) than in plumage, and that yellow coloration is more common than red. We also show that there is no simple, general association between the availability of carotenoids in the diet and the overall use of carotenoid-based coloration. However, when we look at plumage coloration separately from bare part coloration, we find there is a robust and significant association between diet and plumage coloration, but not between diet and bare part coloration. Similarly, when we look at yellow and red plumage colours separately, we find that the association between diet and coloration is typically stronger for red coloration than it is for yellow coloration. Finally, when we build multivariate models to explain variation in each type of carotenoid-based coloration we find that a variety of life history and ecological factors are associated with different aspects of coloration, with dietary carotenoids only being a consistent significant factor in the case of variation in plumage. All of these results remain qualitatively unchanged irrespective of the phylogeny used in the analyses, although in some cases the precise life history and ecological variables included in the multivariate models do vary. Taken together, these results indicate that the predicted link between carotenoid coloration and diet is idiosyncratic rather than general, being strongest with respect to plumage colours and weakest for bare part coloration. We therefore suggest that, although the carotenoid-based bird plumage may a good model for diet-mediated signalling, the use of carotenoids in bare part pigmentation may have a very different functional basis and may be more strongly influenced by genetic and physiological mechanisms, which currently remain relatively understudied. PMID- 16313467 TI - Medicago-Sinorhizobium symbiotic specificity evolution and the geographic expansion of Medicago. AB - The legume genus Medicago interacts with soil bacteria commonly referred to as rhizobia, in a nitrogen fixing symbiosis. We analysed the diversity of symbiotic association specificity among the two organisms, and its evolution in the plant genus. Nitrogen fixation tests and molecular phylogenetic reconstructions revealed that the genus Medicago includes more symbiotic specificity groups than previously suggested and that plant specificity is highly unstable and has repeatedly switched along the diversification of this genus. A phylogenetic analysis including geographical data shows that bacterial geographical diversity distribution has a strong influence on the geographic distribution of plant species and their ability to colonize new areas. Multiple other modifications of specificity occurred along the diversification of the genus, presumably due to selection for specialization to a single bacterial biovar. Codivergence between plants and bacteria may also have taken place. PMID- 16313468 TI - The importance of phylogenetic scale in tests of Bergmann's and Rapoport's rules: lessons from a clade of South American lizards. AB - We tested for the occurrence of Bergmann's rule, the pattern of increasing body size with latitude, and Rapoport's rule, the positive relationship between geographical range size and latitude, in 34 lineages of Liolaemus lizards that occupy arid regions of the Andean foothills. We tested the climatic-variability hypothesis (CVH) by examining the relationship between thermal tolerance breadth and distribution. Each of these analyses was performed varying the level of phylogenetic inclusiveness. Bergmann's rule and the CVH were supported, but Rapoport's rule was not. More variance in the data for Bergmann's rule and the CVH was explained using species belonging to the L. boulengeri series rather than all species, and inclusion of multiple outgroups tended to obscure these macroecological patterns. Evidence for Bergmann's rule and the predicted patterns from the CVH remained after application of phylogenetic comparative methods, indicating a greater role of ecological processes rather than phylogeny in shaping the current species distributions of these lizards. PMID- 16313469 TI - Are hybridogenetic complexes structured by habitat in water frogs? AB - The success and the evolutionary fate of hybridogenetic lineages are explained by both a generalistic heterosis hypothesis and an alternative hypothesis, the habitat segregation hypothesis. Because such hypotheses have rarely been tested at the level of whole habitats, our aim was to compare performances of two taxa within a hybridogenetic complex across diverse natural habitats. We took advantage of the waterfrog hybridogenetic complex (Rana esculenta and R. lessonae) by rearing tadpoles in natural contrasted habitats by means of enclosure experiments. We also monitored the frequency of each taxon in the waterfrog assemblages that naturally breed in the studied ponds. The hybridogenenetic taxon showed no evidence of broader tolerance as growth, development and physiology strongly varied in response to environmental heterogeneity. Our study reveals a differential success of the hybridogenetic taxon and its sexual host among environments. Moreover, hybridogenetic taxa rarely dominated the sexual species in natural assemblages. Consequently, our results show that the generalistic model does not explain the success of hybridogenetic lineages, but rather support the habitat segregation, among other alternative concepts. PMID- 16313470 TI - Adaptive speciation when assortative mating is based on female preference for male marker traits. AB - Adaptive speciation occurs when frequency-dependent ecological interactions generate conditions of disruptive selection to which lineage splitting is an adaptive response. Under such selective conditions, evolution of assortative mating mechanisms enables the break-up of the ancestral lineage into diverging and reproductively isolated descendent species. Extending previous studies, I investigate models of adaptive speciation due to the evolution of indirect assortative mating that is based on three different mating traits: the degree of assortativity, a female preference trait and a male marker trait. For speciation to occur, linkage disequilibria between different mating traits, e.g. between female preference and male marker traits, as well as between mating traits and the ecological trait, must evolve. This can lead to novel speciation scenarios, e.g. when reproductive isolation is generated by a splitting in the degree of assortativeness, with one of the emerging lineages mating assortatively, and the other one disassortatively. I investigate the effects of variation in various model parameters on the likelihood of speciation, as well as robustness of speciation to introducing costs of assortative mating. Even though in the models presented speciation requires the genetic potential for strong assortment as well as rather restrictive ecological conditions, the results show that adaptive speciation due to the evolution of assortative mating when mate choice is based on separate female preference and male marker traits is a theoretically plausible evolutionary scenario. PMID- 16313471 TI - Stress avoidance in a common annual: reproductive timing is important for local adaptation and geographic distribution. AB - Adaptation to local environments may be an important determinant of species' geographic range. However, little is known about which traits contribute to adaptation or whether their further evolution would facilitate range expansion. In this study, we assessed the adaptive value of stress avoidance traits in the common annual Cocklebur (Xanthium strumarium) by performing a reciprocal transplant across a broad latitudinal gradient extending to the species' northern border. Populations were locally adapted and stress avoidance traits accounted for most fitness differences between populations. At the northern border where growing seasons are cooler and shorter, native populations had evolved to reproduce earlier than native populations in the lower latitude gardens. This clinal pattern in reproductive timing corresponded to a shift in selection from favouring later to earlier reproduction. Thus, earlier reproduction is an important adaptation to northern latitudes and constraint on the further evolution of this trait in marginal populations could potentially limit distribution. PMID- 16313472 TI - Life-history invariants with bounded variables cannot be distinguish from data generated by random processes using standard analyses. AB - A dimensionless approach to the study of life-history evolution has been applied to a wide variety of variables in the search for life-history invariants. This approach usually employs ordinary least squares (OLS) regressions of log transformed data. In several well-studied combinations of variables the range of values of one parameter is bounded or limited by the value of the other. In this situation, the null hypothesis normally applied to regression analysis is not appropriate. We generate the null expectations and confidence intervals (CI) for OLS and reduced major axis (RMA) regressions using random variables that are bounded in this way. Comparisons of these CI show that, for log-transformed data, the patterns generated by random data and those predicted by life history invariant theory often could not be distinguished because both predict a slope of 1. We recommend that tests based on the putative invariant ratios and not the correlations between the two variables be used in the exploration of life-history invariants using bounded data. Because empirical data are often not normally distributed randomization test may be more appropriate than standard statistical tests. PMID- 16313473 TI - A genetic map of Cottus gobio (Pisces, Teleostei) based on microsatellites can be linked to the physical map of Tetraodon nigroviridis. AB - To initiate QTL studies in the nonmodel fish Cottus gobio we constructed a genetic map based on 171 microsatellite markers. The mapping panel consisted of F1 intercrosses between two divergent Cottus lineages from the River Rhine System. Basic local alignment search tool (BLAST) searches with the flanking sequences of the microsatellite markers yielded a significant (e < 10(-5)) hit with the Tetraodon nigroviridis genomic sequence for 45% of the Cottus loci. Remarkably, most of these hits were due to short highly conserved noncoding stretches. These have an average length of 40 bp and are on average 92% conserved. Comparison of the map locations between the two genomes revealed extensive conserved synteny, suggesting that the Tetraodon genomic sequence will serve as an excellent genomic reference for at least the Acanthopterygii, which include evolutionarily interesting fish groups such as guppies (Poecilia), cichlids (Tilapia) or Xiphophorus (Platy). The apparent high density of short conserved noncoding stretches in these fish genomes will highly facilitate the identification of genes that have been identified in QTL mapping strategies of evolutionary relevant traits. PMID- 16313474 TI - Apoptosis: a basic biological phenomenon with wide-ranging implications in human disease. AB - Apoptosis is a highly regulated process of cell deletion and plays a fundamental role in the maintenance of tissue homeostasis in the adult organism. Numerous studies in recent years have revealed that apoptosis is a constitutive suicide programme expressed in most, if not all cells, and can be triggered by a variety of extrinsic and intrinsic signals. Many human diseases can be attributed directly or indirectly to a derangement of apoptosis, resulting in either cell accumulation, in which cell eradication or cell turnover is impaired, or cell loss, in which the apoptotic programme is inadvertently triggered. In addition, defective macrophage engulfment and degradation of cell corpses may also contribute to a dysregulation of tissue homeostasis. An increased understanding of the signalling pathways that govern the execution of apoptosis and the subsequent clearance of dying cells may thus yield novel targets for therapeutic intervention in a wide range of human maladies. PMID- 16313475 TI - Resistin gene 3'-untranslated region +62G-->A polymorphism is associated with hypertension but not diabetes mellitus type 2 in a German population. AB - OBJECTIVES: Resistin, a peptide hormone produced by adipocytes, has been associated with diabetes mellitus type 2 (DM-2) in some rodent models. In humans the exact function of resistin remains unknown. Some, but not all studies have found associations between polymorphisms in the resistin gene with DM-2. Recently a 3'-untranslated region +62G-->A polymorphism of the resistin gene has been associated with decreased risk for DM-2 and for hypertension in diabetics in a Chinese population. Purpose of the present study was to examine for the first time in a German Caucasian population the possible association between this polymorphism and DM-2, hypertension, lipoprotein levels, resistin levels as well as atherosclerosis. DESIGN, SETTING AND SUBJECTS: A total of 818 subjects participated in the study. The presence of the +62G-->A polymorphism of the resistin gene was investigated using polymerase chain reaction-restriction fragment length polymorphism in 384 subjects with DM-2 [224 men, 160 women, age 63.4 +/- 10.6 years, body mass index (BMI) 28.7 +/- 5.1 kg m(-2)] and in 434 nondiabetic age- and sex-matched control subjects (248 men, 186 women, age 64.4 +/- 6.5 years, BMI 26.5 +/- 3.7 kg m(-2)). RESULTS: Thirty-four subjects were found to be carrying the +62G-->A polymorphism in the control and 24 in the diabetic group (allelic frequencies 4% and 3.2% respectively). Subjects with DM-2 were not found to have a different frequency of the genotypes (93.75% and 6.258%, for GG:GA/AA respectively) than the control subjects (92.2% and 7.8% for GG:GA/AA respectively) (OR 0.75, 95% CI 0.44-1.3, P = 0.31). In the total cohort, carriers of the A allele had a higher prevalence of hypertension (OR 1.82, 95% CI 1.03 3.21, P = 0.039). When analysed separately, the control group showed a strong association between the presence of the A allele and hypertension (OR 2.92, 95% CI 1.38-6.15, P = 0.005), whilst no such association could be established in the diabetic group (OR 1.05, 95% CI 0.43-2.54, P = 0.92). Multiple regression analysis confirmed that the presence of the A variant is associated with hypertension in control but not in diabetic subjects, independent of age and BMI. The polymorphism had no significant influence on the presence of atherosclerotic disease, BMI, and on triglyceride, HDL and LDL cholesterol levels, both, in the control and the diabetic groups. There was no difference in the serum resistin levels between the 62G-->A variant carriers and noncarriers. CONCLUSIONS: In conclusion, the present data suggest that in a German Caucasian population the +62G-->A polymorphism of the resistin gene is associated with hypertension but not with DM-2. PMID- 16313476 TI - Serum gamma-glutamyltransferase and risk of type 2 diabetes mellitus in men and women from the general population. AB - OBJECTIVES: To examine gender-specific associations between gamma glutamyltransferase (GGT) and incident type 2 diabetes mellitus in a representative population-based sample in Germany. DESIGN: Prospective population based study. METHODS: The study was based on 1851 men and 1836 women (aged 25-64 years) who participated in the first Monitoring Trends and Determinants on Cardiovascular Diseases (MONICA) Augsburg Survey 1984/1985, and who were free of diabetes at baseline. Incident cases of type 2 diabetes were assessed using follow-up questionnaires in 1987/1988, 1997/1998 and 2002/2003 and were validated with medical records. Gender-specific hazard ratios (HRs) were estimated from Cox proportional hazard models. RESULTS: A total of 172 cases of incident type 2 diabetes amongst men and 109 amongst women were registered during a mean follow up period of 14.7 years. In both sexes the risk of type 2 diabetes increased with increasing levels of serum GGT. After multivariable adjustment HRs for incident type 2 diabetes across GGT categories (<25th, <50th, <75th, <87.5th and > or =87.5th percentiles) were 1.0, 1.81, 2.37, 3.41 and 4.24 (P-value for trend <0.0001) in men and 1.0, 1.42, 1.48, 1.95 and 2.41 (P-value for trend 0.0179) in women. Obesity appeared to be more strongly associated with type 2 diabetes in women with GGT equal or greater than the median compared to women with GGT below the median. However, in men the association between obesity and type 2 diabetes was almost identical in the two groups. CONCLUSIONS: The GGT is an important predictor for incident type 2 diabetes in men and women from the general population. PMID- 16313477 TI - Homocysteine levels are not associated with cardiovascular autonomic function in elderly Caucasian subjects without or with type 2 diabetes mellitus: the Hoorn Study. AB - OBJECTIVE: Homocysteine and cardiovascular autonomic function are both predictors of cardiovascular disease and death, particularly in patients with diabetes. The mechanism by which homocysteine causes disease is unknown. The objective of our study was to determine whether hyperhomocysteinaemia is associated with impaired cardiovascular autonomic function in an age-, sex-, and glucose tolerance stratified sample of an elderly Caucasian population. METHODS: We studied 609 subjects, 252 with normal glucose metabolism, 173 with impaired glucose metabolism, and 184 with type 2 diabetes. Cardiac cycle duration (RR interval) and continuous finger arterial pressure were measured under three conditions: during (i) spontaneous breathing, (ii) six deep breaths over 1 min, and (iii) an active change in position from lying to standing. From these readings, 10 parameters of autonomic function were assessed (three Ewing tests, six heart rate variability tests and one test of baroreflex sensitivity). These 10 measurements were summarized in a single cardiovascular autonomic dysfunction score (CADS). RESULTS: Comparing values of autonomic function measures in the lowest versus the highest quartile of homocysteine revealed no significant association between homocysteine level and autonomic function in the whole study group, nor in the individual glucose tolerance groups. Multiple adjustment for age, sex, waist-to hip ratio, serum creatinine, use of antihypertensives and fasting insulin, confirmed this result. We found no evidence of effect modification of glucose tolerance status on the association between homocysteine and autonomic dysfunction (P for interaction for CADS = 0.79). CONCLUSIONS: There is no evidence for an association between homocysteine levels and cardiovascular autonomic function in either diabetic or nondiabetic subjects. Cardiovascular autonomic dysfunction does not help explain why hyperhomocysteinaemia is related to cardiovascular mortality. PMID- 16313478 TI - Effects of 17beta-oestradiol plus different doses of drospirenone on adipose tissue, adiponectin and atherogenic metabolites in postmenopausal women. AB - OBJECTIVE: To investigate how variation in the dose of the progestogen influence the impact of 17beta-oestradiol plus drospirenone (DRSP) treatment on adipose tissue and its secretor function with direct implications for atherogenic metabolites. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Primary care, single study site. SUBJECTS: A total of 240 healthy postmenopausal women 53-65 years old, 178 completer. INTERVENTION: Daily treatment with 1 mg 17beta-oestradiol plus 1, 2, or 3 mg DRSP, or placebo for 2 years. MAIN OUTCOME MEASURES: Absolute changes in central (CFM) and peripheral fat mass (PFM; dual energy X-ray absorptiometry, DEXA), adipokines [interleukin (IL)-6 and adiponectin], atherogenic metabolites [triglycerides, high-density lipoprotein cholesterol (HDL-C), glucose] and blood pressure. RESULTS: Oestradiol plus 1 mg DRSP evoked significant decreases in CFM and the CFM/PFM ratio from baseline. These benefits virtually decreased with increasing dose of DRSP confounded by dose-dependent increases in CFM and PFM in smokers (P-value for trends <0.001), in whom the increases in bioavailable oestradiol were half of that in nonsmokers (P < 0.001). Treatment with 3 mg DRSP induced decreases in serum adiponectin by month 6 (P < 0.05), which persisted in nonsmokers only and led to significant increases in glucose and triglycerides and decreases in HDL-C (P < 0.05). Adiponectin in smokers normalized by the end of the study parallel with the increases in body fat mass. CONCLUSIONS: Interactions of the sex steroids with adipose tissue and its secretor function are important determinants of the overall impact of hormone therapy on cardiovascular risk. A DRSP dose up to 2 mg does not seem to exert adverse effects when combined with 1 mg 17beta-oestradiol. PMID- 16313479 TI - Bone mass in young adults: relationship with gender, weight and genetic factors. AB - OBJECTIVES: To determine the relationship of the bone mass attained in young adults with anthropometric and genetic factors. DESIGN: Cross-sectional study of normal individuals. METHODS: We studied 341 healthy subjects between 22 and 45 years of age. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DXA) and correlated with body weight, height and nine polymorphisms in six genes involved in sex steroid metabolism (17-hydroxylase, aromatase and 5-reductase) and activity (oestrogen receptors (ER)-alpha and beta, and androgen receptor). RESULTS: The BMD was higher in men than in women (spine: 1.048 +/- 0.120 vs. 1.034 +/- 0.112; hip: 0.907 +/- 0.131 vs. 0.822 +/- 0.104 g cm(-2), P < 0.001). However, the difference was due, at least in part, to the larger body size in men and diminished markedly after height adjustment. There was a negative correlation between age and hip BMD. Body weight was the single most influential factor on spine and hip BMD in both sexes, explaining 8 9% of BMD variance. Amongst the genetic factors studied, a common CA repeat polymorphism in ER-beta showed a significant association with BMD in women (P = 0.03 at the spine, and 0.008 at the hip). The relationship between ER-beta genotype and BMD persisted after adjustment by body weight and age, explaining a further 2-3% of BMD variance. Allelic variants of other genes studied were not related with BMD. CONCLUSIONS: Body weight and allelic variants of ER-beta are associated with BMD in young adults. PMID- 16313480 TI - Heparin-induced thrombocytopenia from venous thromboembolism treatment. AB - OBJECTIVES: We aimed to characterize the clinical experiences of patients in whom heparin-induced thrombocytopenia (HIT) complicated heparin therapy for venous thromboembolism (VTE) and who switched to argatroban. DESIGN: A retrospective analysis of previously reported prospective, multicentre, historical-controlled Argatroban-911 and Argatroban-915 studies of argatroban therapy in HIT. SETTING: Inpatient. SUBJECTS: Patients (n = 145) administered heparin for VTE and who developed HIT were identified. INTERVENTIONS: Patients were treated with argatroban 2 mcg kg(-1) min(-1) for up to 14 days, adjusted to maintain activated partial thromboplastin times 1.5 to three times baseline. Patient characteristics, anticoagulation and outcomes were summarized. The primary end point was a composite of death, amputation, or new thrombosis within 37 days of argatroban initiation. RESULTS: During heparin therapy, platelet counts decreased (mean +/- SD nadir: 78 +/- 67 x 10(9) L(-1)), and 75 (52%) patients developed thrombosis. After heparin was discontinued, patients received argatroban (mean dose 2.1 +/- 1.2 mcg kg(-1) min(-1)) for 6.8 +/- 4.3 days. By day 6 of argatroban therapy, the mean platelet count rose to >150 x 10(9) L(-1). The primary end point occurred in 41 (28.3%) patients (values of 26-44% are reported for argatroban therapy of HIT from any heparin indication). Seventeen (11.7%) patients, including 12 who had also experienced thrombosis whilst on heparin, developed new thrombosis after argatroban initiation, typically on the day argatroban was discontinued or later (n = 10). Seven (4.8%) patients experienced major bleeding. CONCLUSIONS: For VTE patients with HIT, argatroban provides effective anticoagulation, with outcomes comparable with those reported for other argatroban-treated HIT patients. New thrombosis in this setting occurred most often in patients with existing HIT-associated thrombosis, before HIT recognition or either at/after argatroban discontinuation. PMID- 16313481 TI - HHV-6-related acute liver failure in two immunocompetent adults: favourable outcome after liver transplantation and/or ganciclovir therapy. AB - Fulminant hepatitis of unknown origin remain a significant cause of mortality, for which liver transplantation is often considered as the only therapeutic option. In retrospective studies, human herpesvirus 6 (HHV-6) infections have been associated with such diseases, but the diagnosis of HHV-6 infection of the liver is rarely established during the acute phase of liver failure. Using real time polymerase chain reaction (PCR), we diagnosed two cases of severe acute liver failure (ALF) related to HHV-6 occurring in immunocompetent young adults. Both cases had a favourable outcome, one after valganciclovir therapy, one after liver transplantation associated with ganciclovir. Viral origin was evidenced in each case by the detection of high amounts of HHV-6 DNA in liver tissue by the PCR assay. The decrease of intrahepatic viral load after therapeutic intervention was also monitored by quantitative PCR and paralleled in the two cases the clinical improvement. Diagnosis of HHV-6 infection must be systematically evoked in case of unexplained ALF, since it might lead to specific therapeutic interventions, in addition of liver transplantation. PMID- 16313482 TI - A HIPS-mimic. PMID- 16313483 TI - TB or TB plus vasculitis: Occam versus Hickam. PMID- 16313485 TI - High iron stores and cardiovascular disease in systemic lupus erythematosus. PMID- 16313487 TI - Atomic force microscopic analysis of hydrogen peroxide bleached kraft northern black spruce fibres. AB - Hydrogen peroxide is a potent, relatively inexpensive oxidant that chemically degrades chromophoric components in pulps and textiles. Oxidation of cellulose is a byproduct of this process step that decreases the tensile strength of individual fibres. The residence time of pulp in the bleaching reactor must be optimized to achieve the desired brightness and minimizing fibre degradation. To evaluate the impact of peroxide bleaching at the microfibrillar level, a single black spruce tree was chosen and kraft pulped. Peroxide bleaching was conducted via benchtop polyethylene bag bleaching in a temperature-controlled waterbath. Atomic force microscopy (AFM) topographical images acquired before and after the bleaching step show dramatic changes in fibre structure consistent with delignification and defects in the surface topography. This was further verified by X-ray work at Brookhaven National Laboratory, NY, U.S.A. PMID- 16313488 TI - Quantitative analysis of keratin filament networks in scanning electron microscopy images of cancer cells. AB - The keratin filament network is an important part of the cytoskeleton. It is involved in the regulation of shape and viscoelasticity of epithelial cells. The morphology of keratin networks depends on post-translational modifications of keratin monomers. In-vitro studies indicated that network characteristics, such as filament crosslink density, determines the biophysical properties of the filament network. This report presents a quantitative method for the morphological analysis of keratin filament networks. Visualization of filaments was based on prefixation extraction of epithelial cells and scanning electron microscopy (SEM). SEM images were processed by a skeletonization algorithm to obtain a graph structure that represents individual filaments as well as their connections. This method was applied to investigate the effects of transforming growth factor alpha (TGFalpha) on the morphology of keratin networks in pancreatic cancer cells. TGFalpha contributes to pancreatic cancer progression and activates signalling pathways phosphorylating keratin monomers. Using this new method, a significant alteration to the keratin network morphology could be detected in response to TGFalpha. PMID- 16313489 TI - Morphological, nanomechanical and cellular structural characterization of human hair and conditioner distribution using torsional resonance mode with an atomic force microscope. AB - Characterization of the cellular structure and chemical and physical properties of hair are essential to develop better cosmetic products and advance the biological and cosmetic sciences. Although the morphology of the fine cellular structure of human hair has traditionally been investigated using scanning electron microscopy and transmission electron microscopy, atomic force microscopy can be used for characterization in ambient conditions without requiring specific sample preparations and surface treatment. In this study, the tapping and torsional resonance modes in an atomic force microscope are compared for measurements of stiffness and viscoelastic properties. The materials were mapped using amplitude and phase angle imaging. The torsional resonance mode showed advantages in resolving the in-plane (lateral) heterogeneity of materials. This mode was used for investigating and characterizing the fine cellular structure of human hair. Various cellular structures (such as the cortex and the cuticle) of human hair and fine sublamellar structures of the cuticle, such as the A-layer, the exocuticle, the endocuticle and the cell membrane complex were easily identified. The distribution and thickness of conditioner on the treated hair surface affects the tribological properties of hair. The thickness of the conditioner was estimated using force distance measurements with an atomic force microscope. PMID- 16313490 TI - Transpiration-assisted perfusion fixation provides in situ preservation of developing ray parenchyma cells in Eucalyptus nitens. AB - The technique of animal vascular perfusion fixation was adapted for in situ fixation of the fragile and difficult to access cells of the ray parenchyma system in stems of 10-year-old Eucalyptus nitens trees. In situ fixation enabled tissue to be safely dissected for histological processing without risk of damage to microstructure or initiation of wound response. Fixative was perfused through the active vascular system under hydrostatic and transpiration pressure directly to vessel-associated ray cells. Diffusion from vessels allowed fixative to access nonvessel-associated ray cells. Acrolein was included to aid diffusion fixation and safranin dye was included to define fixed regions within the xylem. Sections prepared for light and electron microscopy from samples cut from regions showing intense safranin staining showed good microstructural preservation and were free of artefacts caused by mechanical injury or wound response. PMID- 16313491 TI - Destruction of polymer growth substrates for cell cultures in two-photon microscopy. AB - The choice of the growth substrate for cell cultures used in fluorescence microscopy is guided by several factors including the type of cells studied and the type of microscopy used. Usually, cells can be cultured on either polymer or glass substrates. One type of polymer, termed Aclar, presents several attractive features: the adhesive properties are better than those of glass, the optical properties are comparable to those of glass, it is biochemically inert, unbreakable, flexible and has a high surface tension, convenient for seeding cells on the cover slip. However, here we show that when imaging with two-photon microscopy, which is based on a femtosecond pulsed laser source, local damage of the Aclar substrate occurs, starting at an average intensity of 10(5) W cm(-2) at the focal point and for exposure times insufficient to cause cell damage. This leads to the appearance of gas bubbles on cultures plated on Aclar cover slips, which perturb the imaging. By contrast, this phenomenon does not occur on borosilicate cover slips, probably because of their different physical (thermal conductivity, absorbance, melting point) and material homogeneity properties. Thus, for cell culture applications using pulsed lasers with high intensities, the use of glass is preferable to Aclar. The results also reveal that substrates can be more susceptible to thermal damage than the cells themselves. PMID- 16313492 TI - Melatonin as a cytoskeletal modulator: implications for cell physiology and disease. AB - The cytoskeleton is a phylogenetically well-preserved structure that plays a key role in cell physiology. Dynamic and differential changes in cytoskeletal organization occur in cellular processes according to the cell type and the specific function. In neurons, microtubules, microfilaments and intermediate filament (IF) rearrangements occur during axogenesis, and neurite formation which eventually differentiate into axons and dendrites to constitute synaptic patterns of connectivity. In epithelial cells, dynamic modifications occur in the three main cytoskeletal components and phosphorylation of cytoskeletal associated proteins takes place during the formation of the epithelial cell monolayer that eventually will transport water. In pathological processes such as neurodegenerative and psychiatric diseases an abnormal cytoskeletal organization occurs. Melatonin, the main product secreted by pineal gland during dark phase of the photoperiod, is capable of influencing microfilament, microtubule and IF organization by acting as a cytoskeletal modulator. In this paper we will summarize the evidence which provides the data that melatonin regulates cytoskeletal organization and we describe recent findings, which indicate that melatonin effects on microfilament rearrangements in stress fibers are involved in the mechanism by which the indole synchronizes water transport in kidney derived epithelial cells. In addition, we review recent data, which indicates that melatonin protects the neuro-cytoskeletal organization from damage caused by free radicals contributing to cell survival, in addition to the already described mechanism elicited by the indole to prevent apoptosis and to scavenge free radicals. Moreover, we discuss the implications of an altered cytoskeletal organization for neurodegenerative and psychiatric illnesses and its re establishment by melatonin. PMID- 16313493 TI - Detection of N-nitrosomelatonin and other N-nitrosotryptophan derivatives by transnitrosation of APF and DAF-2. AB - S-nitrosothiols can be analyzed with some simple detection procedures and this fact strongly accelerated the understanding of the biological impact of S nitrosothiols. Unfortunately, such simple analytic methods are presently missing for low molecular weight N-nitrosotryptophan derivatives like N-nitrosomelatonin (NOMela). Here we demonstrate that commercially available primary aromatic amines, i.e. aminophenylfluorescein (APF) and 4,5-diaminofluorescin (DAF-2), can be used for a quantitative determination of NOMela. Under optimized conditions (e.g. pH 11) of the assays, the lifetime of N-nitrosotryptophan derivatives is largely prolonged and the reactivity of S-nitrosothiols with aromatic amines can be safely ignored. The influence of reactive nitrogen oxide species like N2O3 is additionally limited at the alkaline pH and may be further decreased by working under hypoxic conditions. As a result of these optimal conditions, the APF assay has a detection limit for NOMela of about 25 nm but this assay fails to detect protein-bound N-nitrosotryptophan residues. The DAF-2 assay, however, might be used for a qualitative analysis of such residues. Due to the high efficacy of the APF assay it is safely demonstrated that in regard to peroxynitrite, N2O3 is about 50-fold more effective in nitrosating melatonin at physiological pH. PMID- 16313494 TI - Melatonin increases survival of HaCaT keratinocytes by suppressing UV-induced apoptosis. AB - Melatonin is a potent antioxidant and direct radical scavenger. As keratinocytes represent the major population in the skin and UV light causes damage to these cells, the possible protective effects of melatonin against UV-induced cell damage in HaCaT keratinocytes were investigated in vitro. Cells were preincubated with melatonin at graded concentrations from 10(-9) to 10(-3) m for 30 min prior to UV irradiation at doses of 25 and 50 mJ/cm2. Biological markers of cellular viability such as DNA synthesis and colony-forming efficiency as well as molecular markers of apoptosis were measured. DNA synthesis was determined by [3H]-thymidine incorporation into insoluble cellular fraction, clonogenicity through plating efficiency experiments and apoptosis by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. DNA synthesis experiments showed a strong protective effect by preincubation with melatonin at concentrations of 10(-4) m (P < 0.01) and 10(-3) m (P < 0.001). Additional postirradiation treatment with melatonin showed no increase in the pre UV incubation protective effect. These results indicate that preincubation is a requirement for melatonin to exert its protective effects. The mechanism of melatonin's protective effect (10(-6) to 10(-3) m) includes inhibition of apoptosis as measured by TUNEL assay. Moreover, the biological significance of these effects is supported by clonogenic studies showing a significantly higher number of colonies in cultures treated with melatonin compared to controls. Thus, pretreatment with melatonin led to strong protection against UVB-induced damage in keratinocytes. PMID- 16313495 TI - Preventive effect of melatonin on bleomycin-induced lung fibrosis in rats. AB - Oxidative stress has an important role in the pathogenesis of idiopathic pulmonary fibrosis. Melatonin has direct and indirect free radical-detoxifying activity. The present study investigated whether melatonin treatment attenuates bleomycin-induced lung fibrosis in rats. A group of rats was given one dose of bleomycin while the control animals were given saline. The first dose of melatonin (4 mg/kg/day) was given 2 days before the bleomycin injection. At day 14, fibrotic changes were evaluated using Aschoft's criteria and lung hydroxyproline content. Bleomycin produced a 2.7-fold rise in the fibrosis score that was decreased 65% by melatonin (P < 0.05) and a 1.4-fold increase in hydroxyproline content which was completely prevented by melatonin. Protein carbonyl and thiobarbituric acid reactive substances levels, which were significantly elevated in the bleomycin treated rats, were significantly attenuated by melatonin. Bleomycin administration significantly reduced the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in lung tissue. The reduction in CAT activity was prevented by melatonin but SOD and GSH-Px were not influenced. These results revealed that melatonin may prevent the development of bleomycin-induced lung fibrosis via the repression of protein and lipid peroxidation. PMID- 16313496 TI - Melatonin reduces pancreatic prostaglandins production and protects against caerulein-induced pancreatitis in rats. AB - Melatonin has been used to treat experimental pancreatitis, although not all the drug's therapeutic mechanisms of melatonin have been defined. Prostaglandins (PGs) are proinflammatory mediators that exert their effects mainly locally during inflammatory diseases. The present study was undertaken to examine whether treatment with melatonin influences local PG production. An acute pancreatitis model in male Sprague-Dawley rats (225-275 g) was established by continuously infusing caerulein (15 mg/kg/hr). Mean arterial pressure and pancreatic perfusion were monitored continuously. Melatonin was delivered via the intraperitoneal route at doses of either 2 or 10 mg/kg, 30 min after caerulein injection. Malondialdehyde and glutathione levels of the pancreas and liver and the trypsinogen activation peptide levels in the serum were measured at the end of the experiment (8 hr after infusion of caerulein). Intraperitoneal injection of melatonin (2 and 10 mg/kg) reduced the reduction in systemic arterial pressure and decreased pancreatic perfusion in the rat model of caerulein pancreatitis. Moreover, melatonin treatment changed local PG production toward control level. Higher dose of melatonin was somewhat more effective in preventing the caerulein induced alterations than was the lower dose. PMID- 16313497 TI - Melatonin protects against lipopolysaccharide-induced intra-uterine fetal death and growth retardation in mice. AB - Lipopolysaccharide (LPS) has been associated with adverse developmental outcomes, including intra-uterine fetal death (IUFD) and intra-uterine growth retardation (IUGR). However, the exact mechanism for LPS-induced IUFD and IURD remains unclear. LPS stimulates macrophages to generate reactive oxygen species (ROS). Therefore, we hypothesize that ROS may be involved in LPS-induced IUFD and IURD. Melatonin is a powerful endogenous antioxidant. In this study, we investigated the protective effects of melatonin on LPS-induced IUFD and IURD in ICR mice. All pregnant mice except controls received an intraperitoneal (75 microg/kg, i.p.) injection of LPS on gestational day (gd) 15-17. The experiment was carried out in two different modes. In mode A, the pregnant mice received two doses of melatonin within 24 hr, one (5 or 10 mg/kg) injected immediately after LPS and the other (5 or 10 mg/kg) injected at 3 hr after LPS. In mode B, the pregnant mice were pretreated with 10 mg/kg of melatonin 18 hr before LPS and then received two doses of melatonin in 24 hr, one (10 mg/kg) injected immediately after LPS and the other (10 mg/kg) injected 3 hr after LPS. The number of live fetuses, dead fetuses and resorption sites were counted on gd 18. Live fetuses in each litter were weighed. Crown-rump and tail lengths were examined and skeletal development was evaluated. Results showed that post-treatments with melatonin significantly attenuated LPS-induced IUFD in a dose-dependent manner. Surprisingly, pre- plus post-treatments with melatonin almost blocked LPS-induced IUFD. In addition, both post-treatments and pre- plus post-treatments with melatonin significantly alleviated LPS-induced decreases in crown-rump and tail lengths and reversed LPS induced skeletal developmental retardation. However, melatonin had little effect on LPS-induced decrease in fetal weight. Furthermore, pre- plus post-treatments with melatonin significantly attenuated LPS-induced lipid peroxidation in maternal liver. These results indicate that melatonin protects against LPS induced IURD and IUGR via counteracting LPS-induced oxidative stress. PMID- 16313498 TI - Acutely administered melatonin is beneficial while chronic melatonin treatment aggravates the evolution of TNBS-induced colitis. AB - The aim of this study was to evaluate the effects of melatonin on the inflammatory response and hydroxyproline production in an experimental acute and chronic model of trinitrobenzene sulfonic (TNBS) acid-induced colitis in Wistar rats. In the acute model, melatonin (0.5, 1, and 2 mg/kg, i.p.) was applied 48, 24, and 1 hr prior to the induction of colitis and 24 and 48 hr after; the severity of colitis was less evident in melatonin-treated animals with significant response in the group treated with 2 mg/kg. All doses investigated significantly reduced the myeloperoxidase activity (MPO). In the chronic studies, melatonin (1 and 2 mg/kg, i.p.) was administered daily 24 hr before hapten instillation and for 7 or 21 days after TNBS; melatonin (2 mg/kg) worsened colitis evolution in the 21-day study with a significant increase in MPO activity and tumor necrosis factor-alpha production with respect to TNBS group. Histological slides were in concordance with macroscopic data where areas of extensive necrosis and edema, fibrosis, and absence of regenerated epithelium were observed. Moreover, the hydroxyproline determination, used as indicator of collagen production and fibrosis, also showed a marker increase. The results obtained in this experimental model showed that short-term administration is protective while in the long term it negatively influences evolution of inflammatory colitis; therefore, the immunostimulatory effect of melatonin in some situations when given chronically, such as during inflammatory bowel disease, might lead to negative consequences. PMID- 16313499 TI - Short- and long-term effects of melatonin on myocardial post-ischemic recovery. AB - Melatonin, the chief secretory product of the pineal gland, has been shown to protect the heart against ischemia-reperfusion injury. This was attributed to its free radical scavenging and broad-spectrum antioxidant properties. The possibility that melatonin may act via its receptor and intracellular signaling, has not yet been addressed in this regard. In all previous studies, only the acute effects of melatonin on the heart, were evaluated. The aims of the present study were to: (i) compare the acute and long-term effects of melatonin on infarct size and functional recovery of the ischemic heart, and (ii) evaluate the role of the melatonin receptor in cardioprotection. For evaluation of the short term effects of melatonin on contractile recovery and infarct size, the isolated perfused working rat heart was subjected to 20 min global ischemia or 35 min regional ischemia respectively, and melatonin (25-50 microm) administered either before and during reperfusion, or before ischemia or during reperfusion after ischemia. The melatonin receptor was manipulated using luzindole and N acetyltryptamine. The long-term effects of melatonin were evaluated 24 hr after melatonin administration (2.5 or 5.0 mg/kg, i.p.) or after oral administration for 7 days (20 or 40 microg/mL). Infarct size and mechanical recovery during reperfusion of the working heart were used as endpoints. Melatonin (50 microm), when administered either before and during reperfusion after ischemia or during reperfusion only, significantly improved cardiac output and work performance and reduced infarct size compared with untreated controls. Luzindole (5 microm), a melatonin receptor antagonist, abolished these cardioprotective effects. Long term administration of melatonin (i.p. or orally for 7 days) caused a significant reduction in infarct size of hearts subjected to 35 min regional ischemia. The cardioprotection persisted for 2-4 days after discontinuation of treatment. In summary, the results obtained suggest that melatonin induces short- as well as long-term protection and that the melatonin receptor is also involved in its cardioprotective actions. PMID- 16313500 TI - Immunocytochemical demonstration of day/night changes of clock gene protein levels in the murine adrenal gland: differences between melatonin-proficient (C3H) and melatonin-deficient (C57BL) mice. AB - The circadian system comprises several peripheral oscillators and a central rhythm generator that, in mammals, is located in the suprachiasmatic nucleus of the hypothalamus. Expression of clock genes is a characteristic feature of the central rhythm generator and the peripheral oscillators. With regard to the rhythmic production of glucocorticoids, the adrenal gland can be considered as peripheral oscillator, but little is known about clock gene expression in this tissue. Therefore, the present study investigates the levels of three clock gene proteins PER1, BMAL1 and CRY2 in the murine adrenal cortex and medulla at seven different time points of a 12-hr light/12-hr dark cycle. To determine a potential role of melatonin we compared the patterns of clock gene proteins in the adrenal gland of melatonin-proficient mice (C3H) with those of melatonin-deficient mice (C57BL). In C3H mice, both, the adrenal cortex and medulla displayed day/night variation in PER1-, CRY2- and BMAL1-protein levels. PER1 and CRY2 peaked in the middle of the light phase, whereas BMAL1 peaked in the dark phase. This pattern was also observed in the adrenal medulla of C57BL, but in the adrenal cortex of C57BL clock gene protein levels did not change with time and were consistently lower than in C3H mice. These results support the hypothesis that the adrenal gland is a peripheral oscillator and raise the possibility that melatonin may be involved in the control of clock gene protein levels in the adrenal cortex of mice. PMID- 16313501 TI - Melatonin counteracts inducible mitochondrial nitric oxide synthase-dependent mitochondrial dysfunction in skeletal muscle of septic mice. AB - Mitochondrial nitric oxide synthase (mtNOS) produces nitric oxide (NO) to modulate mitochondrial respiration. Besides a constitutive mtNOS isoform it was recently suggested that mitochondria express an inducible isoform of the enzyme during sepsis. Thus, the mitochondrial respiratory inhibition and energy failure underlying skeletal muscle contractility failure observed in sepsis may reflect the high levels of NO produced by inducible mtNOS. The fact that mtNOS is induced during sepsis suggests its relation to inducible nitric oxide synthase (iNOS). Thus, we examined the changes in mtNOS activity and mitochondrial function in skeletal muscle of wild-type (iNOS(+/+)) and iNOS knockout (iNOS(-/-)) mice after sepsis. We also studied the effects of melatonin administration on mitochondrial damage in this experimental paradigm. After sepsis, iNOS(+/+) but no iNOS(-/-) mice showed an increase in mtNOS activity and NO production and a reduction in electron transport chain activity. These changes were accompanied by a pronounced oxidative stress reflected in changes in lipid peroxidation levels, oxidized glutathione/reduced glutathione ratio, and glutathione peroxidase and reductase activities. Melatonin treatment counteracted both the changes in mtNOS activity and rises in oxidative stress; the indole also restored mitochondrial respiratory chain in septic iNOS(+/+) mice. Mitochondria from iNOS(-/-) mice were unaffected by either sepsis or melatonin treatment. The data suggest that inducible mtNOS, which is coded by the same gene as that for iNOS, is responsible for mitochondrial dysfunction during sepsis. The results also suggest the use of melatonin for the protection against mtNOS-mediated mitochondrial failure. PMID- 16313502 TI - Sustained activation of Akt by melatonin contributes to the protection against kainic acid-induced neuronal death in hippocampus. AB - In the present study, the underlying protective mechanism of melatonin on kainic acid (KA)-induced excitotoxicity was examined in the hippocampus of mice. KA, administered intracerebroventricularly (i.c.v.), induced marked neuronal cell death with concurrent microglial activation and subsequent induction of inducible nitric oxide synthase (iNOS) in the hippocampus. Histopathological analysis demonstrated that melatonin (10 mg/kg), administered 1 hr prior to KA, attenuated KA-induced death of pyramidal neurons in the CA3 region. Melatonin obviously suppressed KA-induced microglial activation and consequent iNOS expression that were determined by increased immunoreactivities of microglial marker OX-6 and iNOS, respectively. Increased phosphorylation of Akt in pyramidal neurons was observed as early as 2 hr after administration of melatonin. Further, melatonin resulted in increased expression of astroglial glial cell line-derived neurotrophic factor (GDNF), which started to appear approximately 6 hr after administration of melatonin. The results of the present study demonstrate that melatonin exerts its neuroprotective action against KA-induced excitotoxicity both through the activation of neuronal Akt and via the direct action on hippocampal neurons and through the increased expression of astroglial GDNF, which subsequently activates neuronal PI3K/Akt pathway. Therefore, the present study suggests that melatonin, pineal secretory product, is potentially useful in the treatment of acute brain pathologies associated with excitotoxic neuronal damage such as epilepsy, stroke, and traumatic brain injury. PMID- 16313503 TI - 1800 MHz electromagnetic field effects on melatonin release from isolated pineal glands. AB - Isolated pineal glands of Djungarian hamsters (Phodopus sungorus) were continuously perifused by Krebs-Ringer buffer, stimulated with the beta adrenergic receptor agonist isoproterenol to induce melatonin synthesis, and exposed for 7 hr to a 1800 MHz continuous wave (CW) or pulsed GSM (Global System for Mobile Communications)-modulated electromagnetic signal at specific absorption rate (SAR) rates of 8, 80, 800, and 2700 mW/kg. Experiments were performed in a blind fashion. Perifusate samples were collected every hour, and melatonin concentrations were measured by a specific radioimmunoassay. Both types of signal significantly enhanced melatonin release at 800 mW/kg SAR, while at 2700 mW/kg SAR, melatonin levels were elevated in the CW, but suppressed in the GSM-exposed pineal glands. As a temperature rise of approximately 1.2 degrees C was measured at 2700 mW/kg SAR, effects at this level are thermal. With regard to radiofrequency electromagnetic fields, the data do not support the 'melatonin hypothesis,' according to which nonthermal exposure suppresses melatonin synthesis. PMID- 16313504 TI - Ultrastructural clues for the protective effect of melatonin against oxidative damage in cerulein-induced pancreatitis. AB - The role of oxidative stress has been evaluated in experimental models of acute pancreatitis (AP). The aim of this study is to investigate the effect of melatonin on the ultrastructural changes in cerulein-induced AP in rats. Acute pancreatitis was induced by two i.p. injections of cerulein at 2-hr intervals (50 microg/kg BW). One group received additionally melatonin (20 mg/kg BW) i.p. before each injection of cerulein. The rats were sacrificed 12 hr after the last injection. Pancreatic oxidative stress markers were evaluated by changes in the amount of lipid peroxides and changes in the antioxidant enzyme levels, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and total glutathione (GSH) levels. Ultrastructural examination was performed using a transmission electron microscope. Formation of numerous, large autophagosomes, mitochondrial damage, dilatation of rough endoplasmic reticulum (RER) and Golgi apparatus, margination and clumping of nuclear chromatin were the major ultrastructural alterations observed in the AP group. Melatonin administration prevented mitochondrial and nuclear changes and dilatation of RER and Golgi apparatus. Rare, small autophagosomes were present within the cytoplasm of some of the acinar cells. Pancreatic damage was accompanied by a significant increase in tissue MDA levels (P < 0.05) and a significant decrease in CAT, SOD, GPx activities and GSH levels (P < 0.005). Melatonin administration significantly reduced MDA levels but increased CAT, SOD, GPx activities and GSH levels (P < 0.005). Melatonin also reduced serum amylase and lipase activities, which were significantly elevated in AP (P < 0.05 and P < 0.005 respectively). These results suggest that oxidative injury is important in the pathogenesis of AP. Melatonin is potentially capable of limiting pancreatic damage produced during AP by protecting the fine structure of acinar cells and tissue antioxidant enzyme activities. PMID- 16313505 TI - Melatonin therapy in fibromyalgia. PMID- 16313506 TI - Building a partnership to evaluate school-linked health services: the Cincinnati School Health Demonstration Project. AB - The Cincinnati School Health Demonstration Project was a 3-year collaboration that evaluated school-linked health services in 6 urban elementary (kindergarten to eighth grade) schools. Partners from the Cincinnati Health Department, Cincinnati Public Schools, Cincinnati Children's Hospital Medical Center, and The Health Foundation of Greater Cincinnati wanted to determine if levels of school linked care made a difference in student quality of life, school connectedness, attendance, emergency department use, and volume of referrals to health care specialists. School nurses, principals and school staff, parents and students, upper-level managers, and health service researchers worked together over a 2.5 year period to learn about and use new technology to collect information on student health, well-being, and outcome measures. Varying levels of school health care intervention models were instituted and evaluated. A standard model of care was compared with 2 models of enhanced care and service. The information collected from students, parents, nurses, and the school system provided a rich database on the health of urban children. School facilities, staffing, and computer technology, relationship building among stakeholders, extensive communication, and high student mobility were factors that influenced success and findings of the project. Funding for district-wide computerization and addition of school health staff was not secured by the end of the demonstration project; however, relationships among the partners endured and paved the way for future collaborations designed to better serve urban school children in Cincinnati. PMID- 16313507 TI - Competitive foods and beverages available for purchase in secondary schools- selected sites, United States, 2004. AB - School Health Profiles is conducted biennially to assess characteristics of school health programs. State and local departments of education and health select either all public secondary schools within their jurisdictions or a systematic, equal-probability sample of public secondary schools to participate in School Health Profiles. At each school, the principal and lead health education teacher were sent questionnaires to be self-administered and returned to the state or local agency conducting the survey. In 2004, a total of 27 states and 11 large urban school districts obtained weighted data from their survey of principals. The findings in this report indicate that the majority of secondary schools in 27 states and 11 large urban school districts allow students to purchase snack foods or beverages from vending machines or at the school store, canteen, or snack bar. The types of competitive foods and beverages available for purchase varied across states and large urban school districts. Overall, fruits or vegetables were less likely to be available for purchase than the other types of foods or beverages. Bottled water and soft drinks, sports drinks, or fruit drinks that are not 100% juice were most likely to be available for purchase. PMID- 16313508 TI - Associations among risk factors, individual resources, and indices of school related asthma morbidity in urban, school-aged children: a pilot study. AB - This paper presents a conceptual model including examples of risk and resource factors associated with indices of school-related asthma morbidity (eg, missed sleep, participation in activities, school absences) in a group of urban, school aged children with asthma from ethnic minority backgrounds. Specifically, the current longitudinal study examines relations between a contextual risk factor (ie, family life stressors), an asthma-related risk factor (ie, asthma symptoms), individual resources (ie, attention, children's problem-solving beliefs, and self esteem), and aspects of asthma morbidity that have been shown to have an impact on children's academic performance. Participants of the study included 31 mother child dyads from low-income, inner-city neighborhoods. Results of hierarchical regression analyses revealed that after controlling for risk factors (ie, asthma symptoms and family life stressors) at baseline, children's individual characteristics (ie, children's problem-solving beliefs and self-esteem) functioned as resource factors for some indices of asthma-related functioning (school absences, participation in activities, and missed sleep) at follow-up (1 year later). Results suggest that contextual and individual risk and resource factors should be further explored in studies including larger samples of urban children with asthma in order to help guide the development of preventive interventions in school-based and health care settings. PMID- 16313509 TI - Bullying perspectives: experiences, attitudes, and recommendations of 9- to 13 year-olds attending health education centers in the United States. AB - Because 1 in 5 elementary school and 1 in 10 middle school students in the United States report being bullied, understanding student perspectives is crucial to prevention. This study solicited opinions of 9- to 13-year-olds regarding the magnitude of, causes of, and remedies for bullying. Data were obtained from 1229 students visiting 11 health education centers in seven states. Students responded via anonymous, electronic keypads. Half the respondents said they have been bullied at least once in a while. When bullied, almost half said they fight back, about a fourth tell an adult, and 20% do nothing; only 8% try to talk to the bully. Nearly two thirds claimed they tell or try to stop bullying when they see it, but 16% do nothing, and 20% join in. Almost three fourths believed bullying is "uncool," yet 42% said they bully at least occasionally. Bullies and victims were more likely to be middle school age. Frequent bullies were more likely to think it is cool, to fight back when bullied, and to join in when others are bullied. Two classes of victims were apparent. Victims who also bully often said that bullying occurs because others are not friendly to bullies or because bullies want to get their way; many also admitted that they do not know how it can be stopped. Victims who do not bully were most likely to do nothing when bullied but to try to stop the bullying of others. Recommendations are given for using these findings to customize curricula or programs. PMID- 16313510 TI - HIV/AIDS knowledge, attitudes, related behaviors, and sources of information among Korean adolescents. AB - To examine HIV/AIDS knowledge, attitudes, related behaviors, and sources of HIV/AIDS information among high school-aged students in South Korea. One thousand and seventy-seven students (586 females and 491 males) from 5 high schools from 5 representative school districts participated in the survey. A self-administered questionnaire measuring knowledge (19 true-false items), attitudes (4 items, 5 point Likert-type scale), sources of information (6 items, yes/no), and sexual behaviors (8 items, yes/no) was utilized. The level of HIV/AIDS knowledge among Korean adolescents was moderate, with the mean scores of 13.93 out of 19 for males and 13.35 for females (p < .01). Attitudes toward persons with HIV/AIDS were negative, with the mean scores of 3.06 for males and 3.09 for females. Of 42 respondents (4.4%) who had engaged in sexual intercourse, 18 (40%) had used condoms. Almost half of the total respondents reported they were not concerned about HIV/AIDS (46.0%), and 94.4% indicated the need for receiving HIV prevention education in the future. The respondents identified TV (52.5%) and school classes (32.1%) as the two major sources of information on HIV/AIDS. Only a few pointed to their parents (1.3%) as a source of information. This preliminary study summarizes the current status of prevention education on HIV/AIDS available in Korea and can provide implications for developing more differentiated intervention strategies specific to culture, age, and gender. PMID- 16313511 TI - Kids who cut: a protocol for public schools. PMID- 16313512 TI - Novel targets for valproic acid: up-regulation of melatonin receptors and neurotrophic factors in C6 glioma cells. AB - Valproic acid (VPA) is a potent anti-epileptic and effective mood stabilizer. It is known that VPA enhances central GABAergic activity and activates the mitogen activated protein kinase-extracellular signal-regulated kinase (MAPK-ERK) pathway. It can also inhibit various isoforms of the enzyme, histone deacetylase (HDAC), which is associated with modulation of gene transcription. Recent in vivo studies indicate a neuroprotective role for VPA, which has been found to up regulate the expression of brain-derived neurotrophic factor (BDNF) in the rat brain. Given the interaction between the pineal hormone, melatonin, and GABAergic systems in the central nervous system, the effects of VPA on the expression of the mammalian melatonin receptor subtypes, MT1 and MT2, were examined in rat C6 glioma cells. The effects of VPA on the expression of glial cell line-derived neurotrophic factor (GDNF) and BDNF were also examined. RT-PCR studies revealed a significant induction of melatonin MT1 receptor mRNA in C6 cells following treatment with 3 or 5 mm VPA for 24 h or 5 mm VPA for 48 h. Western analysis and immunocytochemical detection confirmed that the VPA-induced increase in MT1 mRNA results in up-regulation of MT1 protein expression. Blockade of the MAPK-ERK pathway by PD98059 enhanced the effect of VPA on MT1 expression, suggesting a negative role for this pathway in MT1 receptor regulation. In addition, significant increases in BDNF, GDNF and HDAC mRNA expression were observed after treatment with VPA for 24 or 48 h. Taken together, the present findings suggest that the neuroprotective properties of VPA involve modulation of neurotrophic factors and receptors for melatonin, which is also thought to play a role in neuroprotection. Moreover, the foregoing suggests that combinations of VPA and melatonin could provide novel therapeutic strategies in neurological and psychiatric disorders. PMID- 16313513 TI - Novel role of sphingosine kinase 1 as a mediator of neurotrophin-3 action in oligodendrocyte progenitors. AB - We had found previously that neurotrophin-3 (NT-3) is a potent stimulator of cAMP response element binding protein (CREB) phosphorylation in cultured oligodendrocyte progenitors. Here, we show that CREB phosphorylation in these cells is also highly stimulated by sphingosine-1-phosphate (S1P), a sphingolipid metabolite that is known to be a potent mediator of numerous biological processes. Moreover, CREB phosphorylation in response to NT-3 involves sphingosine kinase 1 (SphK1), the enzyme that synthesizes S1P. Immunocytochemistry and confocal microscopy indicated that NT-3 induces translocation of SphK1 from the cytoplasm to the plasma membrane of oligodendrocytes, a process accompanied by increased SphK1 activity in the membrane fraction where its substrate sphingosine resides. To examine the involvement of SphK1 in NT-3 function, SphK1 expression was down-regulated by treatment with SphK1 sequence-specific small interfering RNA. Remarkably, the capacity of NT-3 to protect oligodendrocyte progenitors from apoptotic cell death induced by growth factor deprivation was abolished by down-regulating the expression of SphK1, as assessed by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Altogether, these results suggest that SphK1 plays a crucial role in the stimulation of oligodendrocyte progenitor survival by NT-3, and demonstrate a functional link between NT-3 and S1P signaling, adding to the complexity of mechanisms that modulate neurotrophin function and oligodendrocyte development. PMID- 16313514 TI - Role of AP-1 in ethanol-induced N-methyl-D-aspartate receptor 2B subunit gene up regulation in mouse cortical neurons. AB - Activator protein 1 (AP-1) has been reported to regulate the gene expression in a wide variety of cellular processes in response to stimuli. In this study, we investigated the DNA-protein binding activities and promoter activity in the N methyl-D-aspartate R2B (NR2B) gene AP-1 site in normal and ethanol-treated cultured neurons. The identity of the AP-1 site as the functional binding factor is suggested by the specific binding of nuclear extract derived from cultured cortical neurons to the labeled probes and the specific antibody-induced supershift. Mutations in the core sequence resulted in a significantly reduced promoter activity and the ability to compete for the binding. Moreover, treatment of the cultured neuron with 75 mm ethanol for 5 days caused a significant increase in the AP-1 binding activity and promoter activity. The AP-1 DNA-binding complex in control and ethanol-treated nuclear extract was composed of c-Fos, FosB, c-Jun, JunD, and phosphorylated CREB (p-CREB). Western blot analysis showed that p-CREB and FosB significantly increased, whereas c-Jun decreased. The DNA affinity precipitation assay indicated that FosB, p-CREB, and c-Jun increased in the AP-1 complex following ethanol treatment. These results suggest that AP-1 is an active regulator of the NR2B transcription and ethanol-induced changes may result at multiple levels in the regulation including AP-1 proteins expression, CREB phosphorylation and perhaps reorganization of dimmers. PMID- 16313515 TI - Dopamine neurons express multiple isoforms of the nuclear receptor nurr1 with diminished transcriptional activity. AB - Nurr1 (NR4A2) is an orphan nuclear receptor required for the development and maintenance of the dopaminergic phenotype in neurons of the ventral midbrain. This study demonstrates that multiple splice variants of nurr1 are produced in rat and human dopamine neurons. Formed by alternative RNA splicing in exon 7, nurr1a has a truncated carboxy-terminus, nurr1b has an internal deletion in the ligand-binding domain and nurr1c, newly identified in this study, has a novel carboxy-terminus produced by a frame shift downstream of the splice junction. Alternative RNA splicing in exon 3 produces the isoform known as the transcriptionally-inducible nuclear receptor (TINUR), lacking the amino-terminus. Nurr2 and the newly identified nurr2c are produced by utilization of both exon 3 and exon 7 alternative splice sites. In rat midbrain, variants other than full length nurr1 constitute 20-35% of NR4A2 transcripts. Transfection studies in dopaminergic SK-N-AS cells demonstrate that nurr1a, nurr1b, nurr1c and TINUR have significantly reduced transcriptional activities compared with full-length nurr1, while nurr2 and nurr2c are inactive. Furthermore, in these experiments, nurr2 and nurr2c both act as dominant negatives. Production of these nurr1 variants in vivo as demonstrated here could represent a novel regulatory mechanism of nurr1 transcriptional activity and therefore, dopaminergic phenotype. PMID- 16313516 TI - Recombinant prion protein induces rapid polarization and development of synapses in embryonic rat hippocampal neurons in vitro. AB - While a beta-sheet-rich form of the prion protein (PrPSc) causes neurodegeneration, the biological activity of its precursor, the cellular prion protein (PrPC), has been elusive. We have studied the effect of purified recombinant prion protein (recPrP) on rat fetal hippocampal neurons in culture. Overnight exposure to Syrian hamster or mouse recPrP, folded into an alpha helical-rich conformation similar to that of PrPC, resulted in a 1.9-fold increase in neurons with a differentiated axon, a 13.5-fold increase in neurons with differentiated dendrites, a fivefold increase in axon length, and the formation of extensive neuronal circuitry. Formation of synaptic-like contacts was increased by a factor of 4.6 after exposure to recPrP for 7 days. Neither the N-terminal nor C-terminal domains of recPrP nor the PrP paralogue doppel (Dpl) enhanced the polarization of neurons. Inhibitors of protein kinase C (PKC) and of Src kinases, including p59Fyn, blocked the effect of recPrP on axon elongation, while inhibitors of phosphatidylinositol 3-kinase showed a partial inhibition, suggesting that signaling cascades involving these kinases are candidates for transduction of recPrP-mediated signals. The results predict that full-length PrPC functions as a growth factor involved in development of neuronal polarity. PMID- 16313517 TI - Opposing effects of phorbol-12-myristate-13-acetate, an activator of protein kinase C, on the signaling of structurally related human dopamine D1 and D5 receptors. AB - The 'cross-talk' between different types of neurotransmitters through second messenger pathways represents a major regulatory mechanism in neuronal function. We investigated the effects of activation of protein kinase C (PKC) on cAMP dependent signaling by structurally related human D1-like dopaminergic receptors. Human embryonic kidney 293 (HEK293) cells expressing D1 or D5 receptors were pretreated with phorbol-12-myristate-13-acetate (PMA), a potent activator of PKC, followed by analysis of dopamine-mediated receptor activation using whole cell cAMP assays. Unpredictably, PKC activation had completely opposite effects on D1 and D5 receptor signaling. PMA dramatically augmented agonist-evoked D1 receptor signaling, whereas constitutive and dopamine-mediated D5 receptor activation were rapidly blunted. RT-PCR and immunoblotting analyses showed that phorbol ester regulated PKC isozymes (conventional: alpha, betaI, betaII, gamma; novel: delta, epsilon, eta, theta) and protein kinase D (PKCmicro) are expressed in HEK293 cells. PMA appears to mediate these contrasting effects through the activation of Ca2+-independent novel PKC isoforms as revealed by specific inhibitors, bisindolylmaleimide I, Go6976, and Go6983. The finding that cross-talk between PKC and cAMP pathways can produce such opposite outcomes following the activation of structurally similar D1-like receptor subtypes is novel and further strengthens the view that D1 and D5 receptors serve distinct functions in the mammalian nervous and endocrine systems. PMID- 16313518 TI - Supersensitivity of P2X receptors in cerebrocortical cell cultures after in vitro ischemia. AB - Neuronally enriched primary cerebrocortical cultures were exposed to glucose-free medium saturated with argon (in vitro ischemia) instead of oxygen (normoxia). Ischemia did not alter P2X7 receptor mRNA, although serum deprivation clearly increased it. Accordingly, P2X7 receptor immunoreactivity (IR) of microtubuline associated protein 2 (MAP2)-IR neurons or of glial fibrillary acidic protein (GFAP)-IR astrocytes was not affected; serum deprivation augmented the P2X7 receptor IR only in the astrocytic, but not the neuronal cell population. However, ischemia markedly increased the ATP- and 2'-3'-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP)-induced release of previously incorporated [3H]GABA. Both Brilliant Blue G and oxidized ATP inhibited the release of [3H]GABA caused by ATP application; the Brilliant Blue G-sensitive, P2X7 receptor mediated fraction, was much larger after ischemia than after normoxia. Whereas ischemic stimulation failed to alter the amplitude of ATP- and BzATP-induced small inward currents recorded from a subset of non-pyramidal neurons, BzATP caused a more pronounced increase in the frequency of miniature inhibitory postsynaptic currents (mIPSCs) after ischemia than after normoxia. Brilliant Blue G almost abolished the effect of BzATP in normoxic neurons. Since neither the amplitude of mIPSCs nor that of the muscimol-induced inward currents was affected by BzATP, it is assumed that BzATP acts at presynaptic P2X7 receptors. Finally, P2X7 receptors did not enhance the intracellular free Ca2+ concentration either in proximal dendrites or in astrocytes, irrespective of the normoxic or ischemic pre-incubation conditions. Hence, facilitatory P2X7 receptors may be situated at the axon terminals of GABAergic non-pyramidal neurons. When compared with normoxia, ischemia appears to markedly increase P2X7 receptor-mediated GABA release, which may limit the severity of the ischemic damage. At the same time we did not find an accompanying enhancement of P2X7 mRNA or protein expression, suggesting that receptors may become hypersensitive because of an increased efficiency of their transduction pathways. PMID- 16313520 TI - Abstracts for The Royal Australian and New Zealand College of Psychiatrists Joint CINP/ASPR Scientific Meeting, Brisbane, Australia, 7-9 December 2005. PMID- 16313519 TI - Proteomic profiling of cerebrospinal fluid identifies biomarkers for amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is characterized by degeneration of motor neurons. We tested the hypothesis that proteomic analysis will identify protein biomarkers that provide insight into disease pathogenesis and are diagnostically useful. To identify ALS specific biomarkers, we compared the proteomic profile of cerebrospinal fluid (CSF) from ALS and control subjects using surface-enhanced laser desorption/ionization-time of flight mass spectrometry (SELDI-TOF-MS). We identified 30 mass ion peaks with statistically significant (p < 0.01) differences between control and ALS subjects. Initial analysis with a rule learning algorithm yielded biomarker panels with diagnostic predictive value as subsequently assessed using an independent set of coded test subjects. Three biomarkers were identified that are either decreased (transthyretin, cystatin C) or increased (carboxy-terminal fragment of neuroendocrine protein 7B2) in ALS CSF. We validated the SELDI-TOF-MS results for transthyretin and cystatin C by immunoblot and immunohistochemistry using commercially available antibodies. These findings identify a panel of CSF protein biomarkers for ALS. PMID- 16313521 TI - Understanding genetic disease in a socio-historical context: a case study of cystic fibrosis. AB - In this article I present a socio-historical analysis of the definition and diagnosis of one particular genetic disease--cystic fibrosis (CF)--in an effort the better to understand its social context both before and after the advent of molecular genetics. I begin my analysis with some background on the history of CF, before moving on to consider the emergence of the notion of classic CF, the development of the sweat test, early approaches to mild or variant forms of CF, the concept of CF as a genetic disease, the concept of CF as a collection of related disorders, and developments in the understanding and diagnosis of CF which came about in the wake of molecular genetics. I highlight a range of technological, professional and patient developments and how these stimulated new research, typologies and clinical tools. I also consider how different notions of CF were mobilised, either to support or undermine a particular approach to diagnosis or treatment, and consider how the dynamic and contextual characteristics of CF were accounted for by scientists and clinicians with an interest in CF. I end by discussing the implications of my analysis for the contemporary sociology of genetics, and related studies in the sociology of medicine more generally. PMID- 16313522 TI - Dynamic professional boundaries in the healthcare workforce. AB - The healthcare professions have never been static in terms of their own disciplinary boundaries, nor in their role or status in society. Healthcare provision has been defined by changing societal expectations and beliefs, new ways of perceiving health and illness, the introduction of a range of technologies and, more recently, the formal recognition of particular groups through the introduction of education and regulation. It has also been shaped by both inter-professional and profession-state relationships forged over time. A number of factors have converged that place new pressures on workforce boundaries, including an unmet demand for some healthcare services; neo-liberal management philosophies and a greater emphasis on consumer preferences than professional-led services. To date, however, there has been little analysis of the evolution of the workforce as a whole. The discussion of workforce change that has taken place has largely been from the perspective of individual disciplines. Yet the dynamic boundaries of each discipline mean that there is an interrelationship between the components of the workforce that cannot be ignored. The purpose of this paper is to describe four directions in which the existing workforce can change: diversification; specialisation and vertical and horizontal substitution, and to discuss the implications of these changes for the workforce. PMID- 16313523 TI - Social capital and social support on the web: the case of an internet mother site. AB - Do virtual communities in cyberspace foster social capital and social support? Using participant observation and discourse analysis, we examine a mothering board on a parent's website and investigate whether social capital was present, and if so, how it was developed and used. We find three main types of communication emerge from our analysis: emotional support, instrumental support- both formal and informal, and community building/protection, all of which contribute to the creation and maintenance of social capital. Additionally, using sampling with replacement, we created a final data set of 180 mothers and report descriptive statistics to identify characteristics of those on the board. PMID- 16313524 TI - Pro-anorexia, weight-loss drugs and the internet: an "anti-recovery" explanatory model of anorexia. AB - This paper explores the online "pro-anorexia" underground, a movement that supports those with anorexia and adopts an "anti-recovery" perspective on the disease. While encouraging a "healthy" diet to sustain an anorexic way-of-life, the movement also recommends the radical use of weight-loss pharmaceuticals to pursue and maintain low body weight, in contrast to their conventional use to treat obesity. Using ethnographic and interview data collected from participants in the "Anagrrl" website and online forum, we analyse the pro-anorexia (or "pro ana") movement in terms of its underlying "explanatory model" of the disease, and contrast it with medical, psychosocial, sociocultural and feminist models that encourage a "normalisation" of body shape and weight. We suggest that for participants in pro-ana, anorexia represents stability and control, and Anagrrl offers support and guidance for those who wish to remain in this "sanctuary". We discuss the pro-anorexia movement's use of the internet to facilitate resistance to medical and social theories of disease, and its subversion of pharmaceutical technologies. PMID- 16313525 TI - The mundane realities of the everyday lay use of the internet for health, and their consequences for media convergence. AB - The internet is now a major source of health information for lay people. Within the medical, sociological and popular literatures there have been three main responses to this development. We classify these as "celebratory", "concerned" and "contingent". This paper falls into the third category and, drawing on techniques of discourse analysis, examines people's accounts of their use of online health resources. It identifies six implicit rules--which we call "rhetorics of reliability"--that people readily draw upon when articulating why they trust some online sources and not others. In addition participants locate their accounts within broader discursive frameworks in order to present themselves as "sensible" users. The article concludes by suggesting that there is an emerging concordance between the lay use of the internet for health and illness and dominant (generally) biomedical conceptions of what constitutes "good quality" health information. PMID- 16313526 TI - Death brokering: constructing culturally appropriate deaths. AB - Death brokering refers to the activities of medical authorities to render individual deaths culturally meaningful. Social scientists and others agree that mortality provokes existential ambiguity in modern life requiring cultural coping mechanisms. In contemporary Western societies, medical professionals have sequestered the dying in institutions, and have classified the causes of death to explain suspicious death. Over the last decades, the institutionalisation of the dying process has been challenged by social movements and the sudden onset of some deaths while forensic medicine has struggled for professional legitimacy in the borderland between mainstream medicine and the legal system. I argue that medical death brokering persists in spite of challenges because medical experts offer increasingly flexible cultural scripts to render the end-of-life socially meaningful while accentuating death's existential ambiguity. Medical professionals help create the ambiguity they promise to resolve, reinforcing the cultural need for more expert death brokering. PMID- 16313527 TI - Experiences of female general practice registrars: are rural attachments encouraging them to stay? AB - OBJECTIVE: To establish whether a six-month rural attachment influenced female GP registrars' future plans to work in a rural area. Secondary aims include establishing the adequacy of postgraduate training in preparing the registrars for the attachment, opinions regarding rural practice and suggestions to improve the attachment. DESIGN: A six-page questionnaire was sent to eligible participants via state branches of General Practice Education Australia. SUBJECTS: Female GP registrars who undertook a six-month rural attachment during 2002. MAIN OUTCOME MEASURE: Whether the six-month rural attachment influenced female registrars to practise in the country. RESULTS: The rural attachment was a positive experience for 82% of participants. One-third were more likely, as a result of the attachment, to practise rurally in the future. In total, 14% were influenced against working rurally as a result of the attachment. Those who had previously resided or studied in a rural area were more likely to plan to work rurally. Level of vocational preparation was adequate for the majority with notable deficits in obstetrics and gynaecological procedural skills. Main negatives associated with the attachment included working long hours and social isolation. Recommendations for change focused on amending these issues along with improved child-care facilities and improved remuneration. CONCLUSION: The rural attachment is a predominantly positive experience for female registrars with the exception of professional and personal hardships associated with relocating to rural practice. The attachment dissuades only a small proportion of its female counterparts, which is promising considering the increasing role of female practitioners in the workforce. PMID- 16313528 TI - Do the learning needs of rural and urban general practitioners differ? AB - INTRODUCTION: The challenges of rural general practice have given rise to a separate rural training stream and a separate rural professional body. The differences are characterised by the nature of the work undertaken by rural GPs and reflected in the continuing medical education topic choices made when surveyed. METHODS: In 2001 a survey was designed and distributed by the Royal Australian College of General Practitioners and Divisions of General Practice in South Australia and Northern Territory. The survey utilised a list of 104 topics. The topic choices of rural and urban GPs were compared. RESULTS: The survey was distributed to approximately 1762 GPs and yielded 578 responses (33%). Rural GPs were more likely to select the following topics: Anaesthetics, Aboriginal Torres Strait Islander health, Population Health, Renal medicine, Cardiology, Teaching skills, Obstetrics, Neonates, Arrhythmias, Fracture management, Tropical medicine and Therapeutics. Urban GPs were more likely to select Menopause, Travel medicine and Palliative care (P < 0.05). DISCUSSION: Many of the areas of difference reflected aspects of rural general practice. There were also many similarities in topic choices between these two groups. PMID- 16313529 TI - Palliative care in the hinterlands: a description of existing services and doctors' attitudes. AB - OBJECTIVE: To describe palliative care services as they exist in the hinterlands (towns away from regional centres) of Midwest New South Wales, including an estimation of the numbers of cases treated by local doctors, and the service they provide to their patients. Generalist doctors' attitudes to palliative care are also explored, as this information is lacking in the literature but is important for service provision. DESIGN: Descriptive survey. SETTING: Rural primary care and district hospitals. PARTICIPANTS: Generalist doctors in hinterland areas. RESULTS: In total, 38% (19/50) of surveys were returned. "Visiting rights" to the district hospital were held by 78.9% of local doctors, and patients are admitted under the care of their own doctor for symptom control and terminal care, 94.1% and 76.5% of the time, respectively. All doctors surveyed perform home visits for terminally ill patients, and 68.4% make themselves routinely available after hours. Doctors surveyed estimated that they managed a mean of 8.4 deaths due to chronic illness in the past 12 months. Most (78.9%) said that they would continue to manage dying patients even if they had a choice, and they feel between moderately and very satisfied that they provide quality care. However, only 21.1% considered their undergraduate training in palliative medicine to be adequate, and all said that they would refer to a specialist service if it were available. CONCLUSIONS: Generalist rural doctors not only treat many dying patients, but also provide a continuity of care that is rarely seen in other settings. Proposals of models for "rural" palliative care should, therefore, take this unique setting into account. One such suggestion is given in this article. PMID- 16313530 TI - Patterns of home and community care service delivery to culturally and linguistically diverse residents of rural Victoria. AB - OBJECTIVE: To describe and compare patterns of Home and Community Care (HACC) utilisation among culturally and linguistically diverse (CALD) people and Australian-born residents of rural Victoria. DESIGN: The HACC Minimum Data Set provides information regarding levels of service provision and coverage in Victoria. Data from January to June 2002 were analysed to provide a profile of client characteristics and service usage in rural Victoria. Patterns of service utilisation were compared with the profile of the CALD population in the 2001 Census. RESULTS: The proportion of CALD residents who are HACC clients is consistent with demographic profiles. However, their extent of service usage is not consistent with patterns of use by Australian-born residents. HACC clients born in non-English-speaking countries, receive 35% less hours of HACC service than their Australian-born counterparts. HACC clients born overseas in English speaking countries receive nine per cent less hours of HACC service than the Australian-born group (F = 8.9, P = 0.00). Both groups of overseas-born clients use a smaller range of HACC services (F = 1.9, P = 0.16). CONCLUSION: Planners and service providers need to monitor levels of HACC service delivery among population groups to ensure that CALD population groups receive equitable levels of HACC services. The HACC Minimum Data Set is one source of data that can assist in this process. PMID- 16313531 TI - Do health and medical workforce shortages explain the lower rate of rural consumers' complaints to Victoria's Health Services Commissioner? AB - OBJECTIVE: To identify which explanations account for lower rural rates of complaint about health services--(i) fear of consequences where there is little choice of alternative provider; (ii) a higher complaint threshold for rural consumers; (iii) lack of access to complaint mechanisms; or (iv) reduced access to services about which to complain. DESIGN: Ecological study incorporating consumer complaint, population and workforce distribution data sources. SETTING: All health care providers practising in Victoria. PARTICIPANTS: De-identified records of all closed consumer complaints made to the Health Services Commissioner, Victoria, between March 1988 and April 2001 by Victorian residents (13 856 records). MAIN OUTCOME MEASURES: Differences in the percentage of under representation in complaint rates in total and for each of four categories of health services providers for different size communities. RESULTS: No consistent relationship was observed between community size and either degree of under representation of complaints against any category of provider, or the proportion of serious or substantial complaints. Rural under-representation was highest (41%) for dentists, the provider category with the lowest proportion working in rural areas (17%), and lowest (18%) for hospitals, with the highest representation in rural areas (28% of beds). More rural complaints were about access issues (10.7% rural and 8.4% metropolitan). CONCLUSIONS: Reduced opportunity to use health services due to rural health and medical workforce shortages was the best-supported explanation for the lower rural complaint rate. Workforce shortages impact on the quality of rural health services and on residents' opportunities to improve their health status. PMID- 16313532 TI - Health needs in rural areas and the efficacy and cost-effectiveness of doctors and nurses. AB - OBJECTIVE: Because of a lack of GPs in rural areas of Greece it is mandatory for junior doctors to offer medical service in those areas for a year. The aim of this study is to determine the possibility of replacement of internships with nurses and to suggest the most cost-effective way of covering health needs in remote areas. DESIGN: Regional survey. SETTING AND PARTICIPANTS: Patients of primary care offices in two remote areas of Crete, Greece within a year. MAIN OUTCOME MEASURES: Comparative analysis of the level of preventive medicine (estimated by questionnaires) and health needs in the two areas. The reasons for visiting medical offices, references rates, percentages of glucose and blood pressure regulation are also studied. RESULTS: Prescription of drugs for chronic diseases and blood pressure counting were the main reasons for office visits (2868/4594). Respiratory track infections (364/4594) follow. Apart from the high percentages of uncontrolled patients with blood pressure (34%) and diabetes mellitus (14%) there is a high percentage of ignorance or wrong opinions concerning preventive medicine, for example only 63% knew the value of a pap test. CONCLUSIONS: More than two-thirds of "medical" visits in rural areas were for acts that nurses could easily do. The easy access to a junior doctor did not promote preventive medicine. Replacement of junior doctors with properly trained nurses cooperating with GPs responsible for greater regions would be more cost effective than junior doctors improving health in rural areas. Legislation should change, mainly with regard to repeat prescriptions, in order to reduce house visits. PMID- 16313533 TI - Retention of allied health professionals in the south-west of Victoria. PMID- 16313534 TI - Establishing successful distributed clinical teaching. PMID- 16313535 TI - Innovative model of maternity care supporting vulnerable rural populations. PMID- 16313536 TI - From undergraduate student to graduate registered nurse: tips for surviving the transition. PMID- 16313540 TI - ACRRM: Committed to the support of rural and remote women doctors. PMID- 16313541 TI - Double or nothing? Treating exacerbations of asthma. PMID- 16313543 TI - Doubling daily inhaled corticosteroid dose is ineffective in mild to moderately severe attacks of asthma in adults. AB - BACKGROUND: Asthma guidelines recommend increasing or doubling inhaled corticosteroid (ICS) dose to treat mild and moderate exacerbations of asthma in adults. AIM: To: (i) compare the effectiveness of doubling existing daily ICS dose (fluticasone) with maintaining usual ICS dose and usual daily ICS dose accompanied by oral steroids (OS) (dexamethasone) during mild and moderately severe exacerbations of asthma in adults; (ii) examine determinants of success and failure; and (iii) compare side-effect profiles. METHODS: A randomized, double-blind, placebo-controlled (double-dummy), triple crossover trial. Participants acted as their own control. Outcome measures included treatment success/failure, peak expiratory flow (PEF) after 7 days therapy or at treatment failure, and side-effects. RESULTS: From 22 participants (nine males and 13 females), 18 pairs of data were available for maintaining usual ICS versus doubling ICS and doubling ICS versus OS, and 19 for maintaining usual ICS versus OS. Median (fifth-95th percentile) age was 46.5 (32-64) years and forced expiratory volume in one second (FEV(1)) 73% (29-97%) predicted. The outcome after doubling ICS was not superior to maintaining usual ICS, with 11 (61%) failures in both arms (P = 0.66). OS, with only 5 (26%) failures, was superior to maintaining usual ICS with 12 (63%) failures (P = 0.04), and to doubling ICS with 5 (28%) versus 11 (61%) failures (P = 0.07). Median PEF (as percentage of run-in best) at end-points were 90.5% (57.1-177.1) for OS, 78.3% (39.5-103.1) for maintaining usual ICS and 77.9 (27.7-110.3) for doubling ICS. Neither gender nor PEF at exacerbation were predictive of failure. Although doubling ICS was not an effective therapy overall, ICS dose at exacerbation were predictive of success in the doubling ICS arm (P = 0.04). Treatment failures when doubling daily ICS dose were more common if achieved fluticasone dose was less than 2000 microg (three of 11, 73%) compared to 2000 microg or greater (eight of eight, 37.5%). Increasing age and the presence of an upper respiratory tract infection (URTI) were predictive of failure with OS. Side-effects were more commonly reported with OS (52.6%) than doubling ICS (42.1%) or maintaining usual ICS (19.1%) with the most common being mood changes (36.8%), sleep disturbance (31.6%) and changes in appetite (26.3%). CONCLUSIONS: Doubling daily ICS dose per se is not effective for the treatment of mild to moderately severe exacerbations of asthma in adults. Success may depend on achieved ICS dose. Oral steroids are effective, but side effects are common. A review of current guidelines may be warranted. PMID- 16313544 TI - Severe community-acquired pneumonia: an Australian perspective. AB - BACKGROUND: Severe community-acquired pneumonia (CAP) is a common disease with a relatively high mortality. The initial treatment is empirical, based on a broad range of potential pathogens. There are minimal published data describing microbiological causes of pneumonia in Australia. AIMS: To describe the aetiology and characteristics of severe CAP in patients requiring intensive care unit (ICU) admission, to identify factors predicting mortality and to audit current practices of investigation and antibiotic management of these patients from an Australian perspective. METHODS: A retrospective analysis of patient case notes was performed for 96 consecutive patients admitted to two ICU with severe CAP. Data recorded included patient demographics, comorbidities, antimicrobial treatment, investigations and outcome (mortality, length of stay). RESULTS: Overall, mortality was 32%. A microbiological diagnosis was made in 46% of patients. The most frequent causative organisms were Streptococcus pneumoniae (13 cases), influenza A (9), Haemophilus influenzae (5) and Staphylococcus aureus (4); aerobic Gram-negative bacilli collectively accounted for five cases. Blood cultures were positive in 20% of patients. Seventy patients (73%) required mechanical ventilation and 61 patients (63%) required inotropic support. Laboratory abnormalities including acute renal failure, metabolic acidosis and coagulopathy were frequent. Factors associated with mortality on multivariate analysis were age, antibiotic administration prior to hospital presentation, delay in hospital antibiotic administration of more than 4 h, and presence of multilobar or bilateral consolidation on chest X-ray. CONCLUSIONS: Severe CAP requiring ICU admission was associated with a mortality rate of 32%, despite appropriate antimicrobial therapy including a beta-lactam and a macrolide antibiotic in most cases. Causative organisms identified were similar to those found in previous studies. High rates of viral causes (28% of identified pathogens) were noted. Low rates of legionellosis and other atypical causes were found, most probably due to a lack of systematic testing for these agents. PMID- 16313545 TI - Percutaneous closure of patent foramen ovale. AB - AIM: To review the safety and efficacy of percutaneous closure of patent foramen ovale (PFO) in symptomatic patients without the use of general anaesthesia or echocardiographic guidance. METHOD: All patients accepted for percutaneous PFO closure by our service from August 2002 to August 2004 were included. Patient demographics and clinical information were obtained from clinical records and by telephone interview. Follow-up outcomes recorded were recurrence of systemic thromboembolism, residual shunt, and change in pattern of migraines. RESULTS: Forty consecutive adult patients, of whom 19 (48%) had an atrial septal aneurysm (ASA), underwent PFO closure using the Amplatzer occluder device with fluoroscopic guidance alone. Their mean age was 45 +/- 10 years (range 23-63 years) and 24 (60%) were male. The indications for closure were ischaemic stroke (n = 26), transient ischaemic attack (TIA) (n = 8), both stroke and TIA (n = 2), refractory hypoxia (n = 2), platypnoea-orthodeoxia (n = 1) and severe migraine with seizures (n = 1). Twenty patients suffered a single neurological event and 16 suffered >1 event, including six with only radiographic evidence of >1 event. Mean procedure time was 17.7 +/- 9.6 min and fluoroscopy time was 6.7 +/- 7.3 min. Implanted device sizes were 18 mm (n = 1), 25 mm (n = 37) and 35 mm (n = 2). In 33 patients, the procedure was performed as a day-case. No complications were encountered. No further neurological events occurred in 39 patients at a mean follow-up time of 11 +/- 7 months (3-25 months) nor was a significant shunt detected in the 34 who underwent follow-up echocardiography. CONCLUSIONS: Percutaneous closure of PFO with or without ASA, under local anaesthesia and without echocardiographic guidance, is a safe and effective procedure which can be performed as a day-case. This has now become our standard clinical practice. PMID- 16313546 TI - Printed patient education interventions to facilitate shared management of chronic disease: a literature review. AB - BACKGROUND: Printed materials for patient self-education are a comparatively low cost, flexible and potentially useful aid for dealing with the growing burden of chronic disease in our society. A comprehensive review of existing trials is required. AIMS: We conducted a comprehensive review which aimed to (i) evaluate the evidence for effectiveness of print-only interventions in increasing patient participation in chronic disease management and (ii) identify disease or intervention characteristics associated with success. METHODS: Medline, the Cochrane Central Register of Controlled Trials, EMBASE and CINAHL were searched for the period 1992 to January 2005 for reports of controlled trials of printed patient materials in chronic disease. The quality of both studies and interventions was assessed. RESULTS: Seven studies were found. They focussed on inflammatory bowel disease, arthritis, asthma and diabetes. Few studies named primary outcomes, continued beyond 6 months or included process evaluations. Few reports gave the behaviour-change rationale of the intervention. Few interventions were reported as having been designed using testing and improvement, psychosocial theory, checks of patient understanding or other patient input. A variety of outcomes was measured. Statistically significant change was found for few. Knowledge improved in three studies, adherence improved in two and quality of life declined in one. CONCLUSIONS: We found that printed interventions as currently designed appear to produce, at best, modest benefits. The design of interventions, studies and study reports all need improvement before decisions can be made about effectiveness and characteristics associated with success. PMID- 16313547 TI - Warfarin induction at 5 mg daily is safe with a low risk of anticoagulant overdose: results of an audit of patients with deep vein thrombosis commencing warfarin. AB - BACKGROUND: Conventionally warfarin therapy is initiated using a loading dose given over several days. Daily international normalised ratio (INR) monitoring is recommended to prevent overdose; however, with a large proportion of patients with deep vein thrombosis now receiving treatment out of hospital daily blood tests are inconvenient. We introduced a low-dose protocol for starting anticoagulant therapy that only required INR testing on days 4 and 6 and audited the results to assess safety and efficacy. METHODS: Two-hundred and forty-eight patients with confirmed deep vein thrombosis were started on warfarin therapy at 5 mg daily for 3 days. INR measurements were taken at day 4 and day 6. RESULTS: Of these patients, 21% had an INR within the therapeutic range on day 4 and 52% had a therapeutic INR on day 6. The risk of overdose was small with only one case with an INR above 4.0 on day 4 and nine cases on day 6. There were no reported cases of bleeding. CONCLUSION: The low-dose protocol with infrequent testing is safe and convenient for outpatient management. However, our results suggest that patients on this protocol take between 6-10 days to achieve a stable INR. PMID- 16313548 TI - Attention deficit disorder: not just for children. AB - Abstract Attention deficit/hyperactivity disorder (ADHD) has long been described in children who demonstrate developmentally inappropriate symptoms of inattention, impulsivity and motor restlessness. In adults, symptoms are known to persist and the validity of adult ADHD as an entity is now recognized. There is an associated high proportion of other serious psychiatric comorbidities, especially substance abuse, mood and anxiety disorders. Advances have been made into the aetiology and management of ADHD. Many of these focus on the dopamine and noradrenaline pathways. PMID- 16313549 TI - Symptomatic hypocalcaemia and renal impairment associated with bisphosphonate treatment in patients with multiple myeloma. AB - We report three cases of severe hypocalcaemia associated with i.v. bisphosphonate treatment in patients with multiple myeloma. All patients had symptomatic hypocalcaemia, including a tonic-clonic seizure and tachyarrhythmia in one case. Two cases were associated with the development of acute renal failure, whereas the third patient had pre-existing renal impairment. We recommend that bisphosphonates be used with caution in patients with myeloma and renal impairment, that vitamin D deficiency be corrected prior to treatment (to reduce the risk of hypocalcaemia) and that serum calcium and renal function be monitored during treatment. PMID- 16313550 TI - Massive thoracic arterial malformation. PMID- 16313551 TI - Early progression to high-grade dysplasia in a patient with ulcerative colitis and primary sclerosing cholangitis. PMID- 16313552 TI - Subacute respiratory failure due to Arnold-Chiari I malformation. PMID- 16313553 TI - Alpha-1 antitrypsin deficiency, emphysema and chronic obstructive pulmonary disease. PMID- 16313554 TI - Ethical issues in the relationships involving medicine and industry. PMID- 16313555 TI - Mortality in mental illness. PMID- 16313556 TI - Is there a place for placebos in clinical practice? PMID- 16313558 TI - Microarray-based survey of a subset of putative olfactory genes in the mosquito Anopheles gambiae. AB - Female Anopheles gambiae mosquitoes respond to odours emitted from humans in order to find a blood meal, while males are nectar feeders. This complex behaviour is controlled at several levels, but is probably initiated by the interaction of various molecules in the antennal sensilla. Important molecules in the early odour recognition events include odourant binding proteins (OBPs), which may be involved in odour molecule transport, odourant receptors (ORs) that are expressed in the chemosensory neurones and odour degrading enzymes (ODEs). To obtain a better understanding of the expression patterns of genes that may be involved in host odour reception in females, we generated a custom microarray to study their steady state mRNA levels in chemosensory tissues, antennae and palps. These results were supported by quantitative RT PCR. Our study detected several OBPs that are expressed at significantly higher levels in antennae and palps of females vs. males, while others showed the opposite expression pattern. Most OBPs are slightly down-regulated 24 h after blood feeding, but some, especially those with higher expression levels in males, are up-regulated in blood-fed females, suggesting a shift in blood-fed females from human host seeking to nectar feeding. PMID- 16313559 TI - AnoBase: a genetic and biological database of anophelines. AB - AnoBase (http://www.anobase.org) is an integrated, relational database of basic biological and genetic data on anopheline species, with a particular emphasis on Anopheles gambiae. It has been designed as an information source and research support tool for the broad vector biology community. Although AnoBase is not a primary genomic database that develops and provides tools to access the genome of the malaria mosquito, it nevertheless contains several sections that offer data of genomic interest such as in situ hybridization images, an integrated gene tool and direct online access to AnoXcel, the proteomic database of An. gambiae. Moreover, AnoBase also contains information on non-gambiae mosquito species and a novel section on studies related to insecticide resistance. PMID- 16313560 TI - The evolution of immune-related genes from disease carrying mosquitoes: diversity in a peptidoglycan- and a thioester-recognizing protein. AB - Adaptive polymorphism may be common in immune system genes as co-evolutionary interactions foster diversity; either through ongoing positive selection (arms races), or balancing selection. DNA sequence diversity in two putative immune system genes was examined in species of the genus Anopheles and from Aedes aegypti. For one gene, encoding the peptidoglycan recognizing protein PGRPLB, there was evidence of purifying selection, suggesting that selection acts to eliminate sequence variation. For another gene, encoding the thioester-containing protein TEP3, higher levels of amino acid replacement were found than would be expected under neutral models of evolution - an indication that this gene has been subject to repeated bouts of positive selection. PMID- 16313561 TI - Cold hardening and transcriptional change in Drosophila melanogaster. AB - Cold hardening treatment - a brief exposure to low temperatures - can protect certain insects against subsequent exposure to temperatures sufficiently low to cause damage or lethality. Microarray analysis to examine the changes in transcript abundance associated with cold hardening treatment (0 degrees C for 2 h followed by 30 min recovery at 25 degrees C) was undertaken in Drosophila melanogaster in order to gain insight into this phenomenon. Transcripts associated with 36 genes were identified, a subset of which appeared to be also differentially expressed after heat shock treatment. Quantitative RT-PCR was used to independently determine transcript abundance of a subset of these sequences. Taken together, these assays suggest that stress proteins, including Hsp23, Hsp26, Hsp83 and Frost as well as membrane-associated proteins may contribute to the cold hardening response. PMID- 16313562 TI - Aedes aegypti phosphohexomutases and uridine diphosphate-hexose pyrophosphorylases: comparison of primary sequences, substrate specificities and temporal transcription. AB - Phosphohexomutases reversibly catalyse the transfer of the phosphate group of a glycosyl phosphate between the C6 and C1 positions, and uridine diphosphate (UDP) hexose pyrophosphorylases catalyse the synthesis of UDP-hexose from uridine triphosphate (UTP) and hexose-1-phosphate. Both enzyme families are essential for nucleoside diphosphate hexose biosynthesis and are therefore critical for various physiological functions in the midgut of mosquitoes after a blood meal. We cloned and sequenced three phosphohexomutase and two UDP-hexose pyrophosphorylase cDNAs from Aedes aegypti. The products of the cDNAs were expressed and substrate specificities were examined. Herein we describe Ae. aegypti phosphoglucomutase 1, phosphoglucomutase 2, phosphoacetylglucosamine mutase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase. Transcripts of the genes expressing the enzymes are constitutively present in all life stages and blood-feeding does not seem to influence transcript abundance. PMID- 16313563 TI - Assessing the odd secondary structural properties of nuclear small subunit ribosomal RNA sequences (18S) of the twisted-wing parasites (Insecta: Strepsiptera). AB - We report the entire sequence (2864 nts) and secondary structure of the nuclear small subunit ribosomal RNA (SSU rRNA) gene (18S) from the twisted-wing parasite Caenocholax fenyesi texensis Kathirithamby & Johnston (Strepsiptera: Myrmecolacidae). The majority of the base pairings in this structural model map on to the SSU rRNA secondary and tertiary helices that were previously predicted with comparative analysis. These regions of the core rRNA were unambiguously aligned across all Arthropoda. In contrast, many of the variable regions, as previously characterized in other insect taxa, had very large insertions in C. f. texensis. The helical base pairs in these regions were predicted with a comparative analysis of a multiple sequence alignment (that contains C. f. texensis and 174 published arthropod 18S rRNA sequences, including eleven strepsipterans) and thermodynamic-based algorithms. Analysis of our structural alignment revealed four unusual insertions in the core rRNA structure that are unique to animal 18S rRNA and in general agreement with previously proposed insertion sites for strepsipterans. One curious result is the presence of a large insertion within a hairpin loop of a highly conserved pseudoknot helix in variable region 4. Despite the extraordinary variability in sequence length and composition, this insertion contains the conserved sequences 5'-AUUGGCUUAAA-3' and 5'-GAC-3' that immediately flank a putative helix at the 5'- and 3'-ends, respectively. The longer sequence has the potential to form a nine base pair helix with a sequence in the variable region 2, consistent with a recent study proposing this tertiary interaction. Our analysis of a larger set of arthropod 18S rRNA sequences has revealed possible errors in some of the previously published strepsipteran 18S rRNA sequences. Thus we find no support for the previously recovered heterogeneity in the 18S molecules of strepsipterans. Our findings lend insight to the evolution of RNA structure and function and the impact large insertions pose on genome size. We also provide a novel alignment template that will improve the phylogenetic placement of the Strepsiptera among other insect taxa. PMID- 16313564 TI - 20-Hydroxyecdysone indirectly regulates Hemolin gene expression in Hyalophora cecropia. AB - Development and innate immune defence are two vital processes that have been demonstrated to use the same or similar molecules and signalling pathways in insects. Hemolin is a moth haemolymph protein belonging to the immunoglobulin superfamily. It is strongly induced upon bacterial infection. However, recent studies indicate a developmental regulation of hemolin. We show that the steroid hormone 20-hydroxyecdysone (20E) can activate the expression of Hyalophora cecropia Hemolin (HcHemolin) in the fat body of diapausing pupae. Using the protein synthesis inhibitor cycloheximide we demonstrate that Hemolin up regulation by 20E requires ongoing protein synthesis. Moreover, 20E enhances transcription of the Hemolin gene in response to bacteria. Comparing the upstream regions of Manduca sexta Hemolin (MsHemolin) and HcHemolin, we identified four putative regulatory sites. Two are putative hormone response elements (HREs), one with an imperfect inverted repeat (HRE-IR) and one with a monomeric site (HRE-M). An additional monomeric hormone receptor site (MRE) is present only in HcHemolin. The third conserved motif is similar to the interferon (IFN) regulatory factor binding element (IRF-E) and IFN-stimulated response element (ISRE). The fourth conserved element is a kappaB motif situated between the Cap-site and the TATA box. Finally, by electrophoresis mobility shift assay we demonstrate that the HRE IR forms specific complexes with nuclear extract proteins of normal pupae that increase after 20E stimulation. PMID- 16313565 TI - Genetic variability in the three genomes of Nasonia: nuclear, mitochondrial and Wolbachia. AB - Nasonia consists of three closely related species of parasitoid wasps that are all infected with the endosymbiotic bacteria Wolbachia, a reproductive parasite common in arthropods. This situation presents the opportunity to compare patterns of variation in three associated genomes, Wolbachia and the nuclear and mitochondrial genomes of its host. Furthermore, although Nasonia wasps are emerging as a model for evolutionary and genetic studies, little is known about their genetic variability. Using amplified fragment length polymorphisms (AFLPs), all three species present a relatively high level of nuclear polymorphism and have different patterns of variation, with one of the species, Nasonia giraulti, being divided into two divergent subgroups. In each species, the mitochondrial pattern of variation is different from the nuclear pattern, possibly due to genetic hitchhiking of the mitochondria during (cytoplasmically inherited) Wolbachia sweeps. Mitochondria in Nasonia show a synonymous substitution rate approximately 10-15-fold higher than nuclear genes, probably reflecting an elevated mitochondrial mutation rate that is among the highest found in insects. Finally, all three species are doubly infected with their own strains of Wolbachia, one each from the two major supergroups (A and B). Sequence analysis reveals that each of the three Nasonia species acquired their A and B bacteria independently by horizontal transfer events from other insects with the exception of B type Wolbachia in N. longicornis and N. giraulti, which were acquired prior to speciation and then codiverged with the host. This represents one of the few clear-cut examples of codivergence of Wolbachia during host speciation. PMID- 16313566 TI - Radiation-induced changes in gene expression in the silkworm revealed by serial analysis of gene expression (SAGE). AB - Serial analysis of gene expression (SAGE) was used to examine the profile of expressed genes during embryonic development in the domesticated silkworm, Bombyx mori, after irradiation with Cobalt-60. A comparison of the SAGE sequence tags derived from irradiated embryos with those from normal embryos revealed 673 differentially expressed genes (P < 0.01 and at least three folds change). Of these, 292 genes were highly expressed in normal embryos and 381 genes were highly expressed in irradiated embryos. These results provide valuable information for understanding the mechanisms of radiation-induced changes in gene expression. In addition, it was noted that the generation of longer cDNA fragments from SAGE tags is an efficient way to identify genes, thereby facilitating the analysis of large numbers of unknown genes. PMID- 16313567 TI - Role of dopachrome conversion enzyme in the melanization of filarial worms in mosquitoes. AB - Melanization is an effective defence reaction of mosquito hosts against invading parasites. In mosquitoes, the biosynthesis of melanin is initiated by the hydroxylation of tyrosine to DOPA by phenoloxidase (PO). DOPA is a branch point of the melanization reaction; it may be oxidized to dopaquinone by PO or be decarboxylated to dopamine by dopa decarboxylase. Further oxidation of dopaquinone by PO produces dopachrome. Dopachrome is then converted to 5, 6 dihydroxyindole by dopachrome conversion enzyme (DCE) to produce melanin. The conversion of dopachrome is a rate-limiting step of the melanization reaction, and the presence of PO and DCE significantly accelerates melanization reactions. In this study, a cDNA encoding DCE was cloned from the mosquito Armigeres subalbatus. Real-time PCR analysis revealed increased transcripts from haemocytes in microfilariae (mf)-inoculated mosquitoes. Gene silencing using double-stranded RNA was used to elucidate the role of DCE in the melanization reaction of parasites in Ar. subalbatus. The levels of both DCE transcripts and protein in gene knockdown mosquitoes were dramatically reduced. Compared with controls, the degree of melanization of mf in DCE-knockdown mosquitoes was significantly decreased. These results suggest that DCE is a critical enzyme that is required for effective melanization immune responses. PMID- 16313568 TI - An unusual distribution of the kdr gene among populations of Anopheles gambiae on the island of Bioko, Equatorial Guinea. AB - In West Africa, Anopheles gambiae exists in discrete subpopulations known as the M and S molecular forms. Although these forms occur in sympatry, pyrethroid knock down resistance (kdr) is strongly associated with the S molecular form. On the island of Bioko, Equatorial Guinea we found high frequencies of the kdr mutation in M form individuals (55.8%) and a complete absence of kdr in the S form. We also report the absence of the kdr allele in M and S specimens from the harbour town of Tiko in Cameroon, representing the nearest continental population to Bioko. The kdr allele had previously been reported as absent in populations of An. gambiae on Bioko. Contrary to earlier reports, sequencing of intron-1 of this sodium channel gene revealed no fixed differences between M form resistant and susceptible individuals. The mutation may have recently arisen independently in the M form on Bioko due to recent and intensive pyrethroid application. PMID- 16313569 TI - Expressed sequence tags from the midgut and an epithelial cell line of Chironomus tentans: annotation, bioinformatic classification of unknown transcripts and analysis of expression levels. AB - Expressed sequence tags (ESTs) were generated from two Chironomus tentans cDNA libraries, constructed from an embryo epithelial cell line and from larva midgut tissue. 8584 5'-end ESTs were generated and assembled into 3110 tentative unique transcripts, providing the largest contribution of C. tentans sequences to public databases to date. Annotation using Blast gave 1975 (63.5%) transcripts with a significant match in the major gene/protein databases, 1170 with a best match to Anopheles gambiae and 480 to Drosophila melanogaster. 1091 transcripts (35.1%) had no match to any database. Studies of open reading frames suggest that at least 323 of these contain a coding sequence, indicating that a large proportion of the genes in C. tentans belong to previously unknown gene families. PMID- 16313570 TI - The expression of the HSP90 gene in response to winter and summer diapauses and thermal-stress in the onion maggot, Delia antiqua. AB - The full-length Hsp90 cDNA in Delia antiqua was cloned and sequenced. The deduced polypeptide comprised 717 amino acid residues, with a molecular mass of 82 140 Da. Summer- and winter-diapauses both elevated HSP90 transcript levels in D. antiqua pupae. Levels gradually increased with time in summer diapausing pupae whereas levels fluctuated in winter diapausing pupae. Cold- and heat-stressing summer- and winter-diapausing individuals further elevated HSP90 expression. mRNA levels gradually increased with time in summer diapausing pupae whereas levels decreased with time after an initial increase in winter diapausing pupae. HSP90 expression was also up-regulated following cold- and heat-stresses in non diapausing pupae. Heat-stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase. These results suggest that the development and physiology of summer- and winter diapauses, as monitored via variation in HSP90 transcript levels, can be substantial different. PMID- 16313571 TI - Characterization of a blood activated chitinolytic system in the midgut of the sand fly vectors Lutzomyia longipalpis and Phlebotomus papatasi. AB - We characterized a cDNA from Phlebotomus papatasi, PpChit1, which encodes a midgut specific chitinase and show the presence of a functional, blood-induced chitinolytic system in sand flies. PpChit1 is detected only in the midgut and is regulated by blood feeding. A recombinant protein (rPpChit1) produced in HEK 293 F cells exhibited a similar activity profile to that found in the native protein against several specific substrates, including an oligomeric glycol chitin and synthetic 4-methyl-umbelliferone labelled substrates. Western blotting showed that the native protein is recognized by mouse polyclonal antibodies against rPpChit1. Additionally, the rPpChit1 and the native chitinase displayed similar retention times in a HPLC size fractionation column. When added to rPpChit1 or to midgut lysates, PpChit1 sera reduced chitinolytic activity by 65-70%. PMID- 16313572 TI - Molecular characterization and evolution of the repeating units of histone genes in Drosophila americana: coexistence of quartet and quintet units in a genome. AB - Quintet and quartet repeating units of the histone genes in Drosophila americana were cloned and characterized. Nucleotide sequence analysis of the units showed that a 3175 bp unit contained the core histone genes but lacked the H1 gene ('quartet unit') while a 5025 bp unit contained all five histone genes ('quintet unit'). Comparative analysis suggested that these repeating units diverged before the separation of D. americana and D. virilis. Multiple forms of H1 genes, differing by 5.8% of amino acids, were found in D. americana. The genomic organization of the histone gene family in D. americana was found to be very similar to that of D. virilis. PMID- 16313574 TI - Entrustability of professional activities and competency-based training. PMID- 16313575 TI - Beyond the 'four Ds of Multiculturalism': taking difference into account in medical education. PMID- 16313576 TI - Conducting ethnography in medical settings. AB - BACKGROUND: Ethnographic research methods have a considerable history of use in medical settings. This paper provides a personal account of conducting ethnography in medical environments and focuses on some of the practical and ethical issues encountered in such settings. DISCUSSION AND CONCLUSION: The paper describes particular issues that arise when undertaking ethnographic work, such as gaining access, recording data, research roles and researching elite groups, and recounts some of the dilemmas posed by doing this type of research in a medical setting. The paper concludes by reflecting on what ethnography might offer to medicine in return for the rich data medicine provides. PMID- 16313577 TI - A practical guide to assessing clinical decision-making skills using the key features approach. AB - AIM: This paper in the series on professional assessment provides a practical guide to writing key features problems (KFPs). Key features problems test clinical decision-making skills in written or computer-based formats. They are based on the concept of critical steps or 'key features' in decision making and represent an advance on the older, less reliable patient management problem (PMP) formats. METHOD: The practical steps in writing these problems are discussed and illustrated by examples. Steps include assembling problem-writing groups, selecting a suitable clinical scenario or problem and defining its key features, writing the questions, selecting question response formats, preparing scoring keys, reviewing item quality and item banking. CONCLUSION: The KFP format provides educators with a flexible approach to testing clinical decision-making skills with demonstrated validity and reliability when constructed according to the guidelines provided. PMID- 16313578 TI - A comprehensive collaborative patient safety residency curriculum to address the ACGME core competencies. AB - CONTEXT: Patient safety currently receives only scant attention in most residency curricula. Safety is a subject that transcends the US Accreditation Council for Graduate Medical Education's 6 core competencies. OBJECTIVE: To design and implement a new patient safety curriculum in collaboration with the Schools of Nursing and Pharmacy, in such a way as to address all 6 competencies. SETTING AND PARTICIPANTS: The curriculum applies to a university-based family medicine residency programme with 45 residents at 5 sites, including urban, suburban and rural sites. CURRICULUM DESIGN: The curriculum includes introductory workshops for faculty and residents, a series of didactic courses, individual portfolios and a series of small group exercises including chart reviews, case presentations and a longitudinal quality improvement project. The activities are run by a multidisciplinary team. OUTCOME MEASURES: Main outcome measures include assessment of resident performance in curriculum activities and in an annual objective structured clinical examination (OSCE) that includes standardised patient interviews, simulations and a written examination. Programme evaluation will include comparison of OSCE performance with that at a neighbouring residency. RESULTS: Residents identified safety problems and system-based solutions using a safety journal. Cases of polypharmacy were identified using journals and chart reviews, and medication changes proposed and discussed. At resident practice sites, residents identified safety priorities based on a staff survey and proposed system-based solutions. Results of the OSCE will be presented elsewhere. CONCLUSIONS: A new patient safety curriculum was successfully introduced into a family medicine residency. The curriculum integrates patient safety into residents' daily activities and incorporates input from the disciplines of nursing and pharmacy so as to help build more effective clinical teams and inculcate a culture of safety. PMID- 16313579 TI - Educating doctors in France and Canada: are the differences based on evidence or history? AB - BACKGROUND: Despite many economic and political similarities between France and Canada, particularly in their health care systems, there are very significant differences in their systems of medical education. AIM: This work aims to highlight the sociohistorical values of each country that explain these differences by comparing the medical education systems of the 2 countries, including medical schools (teachers, funding), key processes (curriculum, student selection) and quality assurance methods. DISCUSSION: In France, means and processes are standardised and defined at a national level. France has almost no national system of assessment of medical schools nor of students. By contrast, Canada leaves medical schools free to design their medical curricula, select students and appoint teachers using their own criteria. In order to guarantee the homogeneity and quality of graduates, the medical profession in Canada has created independent national organisations that are responsible for accreditation and certification processes. Each country has a set of founding values that partly explain the choices that have been made. In France these include equality and the right to receive free education. In Canada, these include equity, affirmative action and market-driven tuition. CONCLUSION: Many of the differences are more easily explained by history and national values than by a robust base of evidence. There is a constant tension between a vision of education promoted by medical educators, based on contextually non-specific ideas such as those found in the medical education literature, and the sociopolitical foundations and forces that are unique to each country. If we fail to consider such variables, we are likely to encounter significant resistance when implementing reforms. PMID- 16313580 TI - The development of medical teachers: an enquiry into the learning histories of 10 experienced medical teachers. AB - AIM: The aim of the study was to explore the different ways in which doctors have learned to teach and train. INTRODUCTION: There is no coherent theory of medical teacher development. Doctors are experts in what they teach; most have had little or no training in how they teach. Research has mostly concentrated on the acquisition and improvement of pedagogical skills by attendance at formal, generally short courses. These may have limited impact. METHODS: We carried out semistructured interviews with 10 experienced medical teachers. A review of the literature had suggested areas to explore. Interviews were transcribed and coded and thematic analysis and grounded theory used as the framework for qualitative analysis. RESULTS: Four areas were identified as important in teacher development: acquisition of educational knowledge and skills; modelling and practice of teaching skills; encouragement and motivation of teachers, and constraints on teaching and learning. DISCUSSION: The results suggest a model for teacher development that begins with doctors as learners, learning to learn and watching teachers teach. They then start to teach, acquiring and practising skills, and subsequently move on to reflect on their teaching. They can be encouraged to teach but may also be prevented from teaching. CONCLUSIONS: This inductive study proposes a model for medical teacher development that attempts to explain how doctors learn to teach and train. More research is needed to clarify the findings. There are implications for faculty development. PMID- 16313581 TI - The interpersonal, cognitive and efficiency domains of clinical teaching: construct validity of a multi-dimensional scale. AB - BACKGROUND: We are unaware of any hypothesis-driven studies showing that teaching assessments are comprised solely of interpersonal and cognitive domains. Moreover, previous teaching assessments have been biased by heterogeneous samples of evaluators. Consequently, we investigated the construct validity of faculty assessments comprised of interpersonal and cognitive domains, utilising evaluations obtained from resident doctors on an internal medicine hospital service. METHODS: A total of 1000 inpatient evaluations were completed on 60 general internal medicine faculty members. Education theory supported a 2 dimensional, 14-item scale. Principal factor analysis was used to explore the scale's dimensionality. Internal reliability and interobserver agreement were determined. Relationships between domains and instructor characteristics were also examined. RESULTS: Principal factor analysis revealed interpersonal, clinical teaching and efficiency domains. Internal reliabilities of all domains are high (alpha > 0.90). Interobserver agreement is good (range 0.64-0.83). In the interpersonal domain there is a trend towards higher scores for lower ranking faculty. Significant findings are higher overall scores in the interpersonal domain (P < 0.001), higher scores for assistant professors in the interpersonal domain (P = 0.008) and higher scores for male than female faculty in the interpersonal (P = 0.041) and clinical teaching (P = 0.008) domains. CONCLUSIONS: Clinical teaching evaluations are reducible to interpersonal, clinical teaching and efficiency domains. Evidence for construct validity includes predicted domains and high internal and interobserver reliabilities. Utilising a homogenous sample of evaluators minimised variance. Interestingly, lower ranking faculty scored higher in the interpersonal domain, suggesting that lower ranking faculty may focus more attention on teaching activities than full professors do. PMID- 16313582 TI - Conditions for successful reflective use of portfolios in undergraduate medical education. AB - AIM: Portfolios are often used as an instrument with which to stimulate students to reflect on their experiences. Research has shown that working with portfolios does not automatically stimulate reflection. In this study we addressed the question: What are the conditions for successful reflective use of portfolios in undergraduate medical education? METHODOLOGY/RESEARCH DESIGN: We designed a portfolio that was aimed at stimulating reflection in early undergraduate medical education, using experiences described in the medical education literature and elsewhere. Conditions for reflective portfolio use were identified through interviews with 13 teachers (mentors), who were experienced in mentoring students in the process of developing their portfolios. The interviews were analysed according to the principles of grounded theory. RESULTS: The conditions for successful reflective use of portfolios that emerged from the interviews fell into 4 categories: coaching; portfolio structure and guidelines; relevant experiences and materials, and summative assessment. According to the mentors, working with a portfolio designed to meet these conditions will stimulate students' reflective abilities. CONCLUSION: This study shows that portfolios are a potentially valuable method of assessing and developing students' reflective skills in undergraduate medical training, provided certain conditions for effective portfolios are recognised and met. Portfolios have a strong potential for enhancing learning and assessment but they are very vulnerable and may easily lead to disappointment. Before implementing portfolios in education, one should first consider whether the necessary conditions can be fulfilled, including an appropriate portfolio structure, an appropriate assessment procedure, the provision of enough new experiences and materials, and sufficient teacher capacity for adequate coaching and assessment. PMID- 16313583 TI - An explorative study into learning on international traineeships: experiential learning processes dominate. AB - OBJECTIVE: To explore the learning processes of undergraduate medical students undertaking international traineeships. METHODS: Semi-structured, in-depth interviews were conducted with 24 undergraduate medical students from Maastricht University Medical School, the Netherlands. The 24 subjects were selected by purposeful sampling. Research methods resembled the grounded theory method. Although the data were initially gathered for an earlier study, the richness of the data enabled renewed scrutiny. RESULTS: The data yielded the following categories concerning the description of the learning processes: motivation, preparation, internal supervisor, external supervisor, assessment, and positive and negative incidents. The majority of supervisors' influences on the learning process were facilitative or negligible. Socio-cultural differences between students and supervisors sometimes appeared to blur productive learning. The greater proportion of the students seemed to learn by 'experiential learning', a smaller proportion learned by 'active learning', and the learning of a very small minority of the students appeared to be 'guided'. DISCUSSION: Although experiential learning seemed to be the dominant learning process, it is not clear whether this is also the most fruitful approach to learning. It could be argued that structuring learning outcomes by a more active learning process might be more effective. To guide the learning process and learning outcomes, it might be advisable to develop guidelines for writing international traineeship reports. Supervision of these reports should be more co-ordinated to prevent arbitrariness and to scaffold active learning. PMID- 16313584 TI - Analysing the concept of context in medical education. AB - BACKGROUND: There is increasing interest in the role of context in medical education, with the conjecture that learning in a clinical context may be helpful for later recall of knowledge. Although this may be true in a general sense, at a closer look it appears that the notion of context is not well substantiated in the medical education literature and that the concept is not clearly defined. Effects of context on learning appear to depend on type of learning task, the relationship or interaction between the context and the learning material, and motivational features of the context. Context is often implicitly regarded as a uniform concept but conceptual analysis shows that a distinction can be made in several dimensions. RESULTS: In this paper, we identify 3 different dimensions of context: a physical dimension, representing the environmental characteristics; a semantic dimension, reflecting how well the context contributes to the learning task, and a commitment dimension, representing the amount of commitment (in terms of motivation and responsibility) that is generated by the context. On these dimensions, context can be ordered from reduced (providing few cues, little meaning, little commitment) to enriched (many cues, much meaning, high commitment). CONCLUSION: This model can serve a dual purpose: first, to disentangle several aspects of educational contexts (e.g. as high in meaning but low in commitment), and second, to provide a theoretical framework to generate research on the influence of different contexts in education on students' learning. PMID- 16313585 TI - A further example of paired-teacher lecturing to link theory to practice. PMID- 16313586 TI - Attitudes of medical students and residents toward multidisciplinary team approach. PMID- 16313587 TI - The voyage of an invasive species across continents: genetic diversity of North American and European Colorado potato beetle populations. AB - The paradox of successful invading species is that they are likely to be genetically depauperate compared to their source population. This study on Colorado potato beetles is one of the few studies of the genetic consequences of continent-scale invasion in an insect pest. Understanding gene flow, population structure and the potential for rapid evolution in native and invasive populations offers insights both into the dynamics of small populations that become successful invaders and for their management as pests. We used this approach to investigate the invasion of the Colorado potato beetle (Leptinotarsa decemlineata) from North America to Europe. The beetles invaded Europe at the beginning of the 20th century and expanded almost throughout the continent in about 30 years. From the analysis of mitochondrial DNA (mtDNA) and amplified fragment length polymorphism (AFLP) markers, we found the highest genetic diversity in beetle populations from the central United States. The European populations clearly contained only a fraction of the genetic variability observed in North American populations. European populations show a significant reduction at nuclear markers (AFLPs) and are fixed for one mitochondrial haplotype, suggesting a single successful founder event. Despite the high vagility of the species and the reduction of genetic diversity in Europe, we found a similar, high level of population structure and low gene flow among populations on both continents. Founder events during range expansion, agricultural management with crop rotation, and selection due to insecticide applications are most likely the causes partitioning genetic diversity in this species. PMID- 16313588 TI - Bayesian inference of evolutionary history from chloroplast microsatellites in the cosmopolitan weed Capsella bursa-pastoris (Brassicaceae). AB - Besides showing an extraordinary degree of phenotypic variability, Capsella bursa pastoris (Brassicaceae) is also one of the world's most common plant species and a serious weed in many countries. We have employed a coalescent-based Bayesian analysis of chloroplast microsatellite data to infer demographic and evolutionary parameters of this species. Two different demographic models applied to data from seven chloroplast microsatellite loci among 59 accessions show that the effective population size of C. bursa-pastoris is very small indicating a rapid expansion of the species, a result that is in accordance with fossil and historical data. Against this background, analysis of flowering time variation among accessions suggests that ecotypic differentiation in flowering time has occurred recently in the species' history. Finally, our results also indicate that mononucleotide repeat loci in the chloroplast genome can deteriorate in relatively short periods of evolutionary time. PMID- 16313589 TI - Rangewide population genetic structure of the African malaria vector Anopheles funestus. AB - Anopheles funestus is a primary vector of malaria in Africa south of the Sahara. We assessed its rangewide population genetic structure based on samples from 11 countries, using 10 physically mapped microsatellite loci, two per autosome arm and the X (N = 548), and 834 bp of the mitochondrial ND5 gene (N = 470). On the basis of microsatellite allele frequencies, we found three subdivisions: eastern (coastal Tanzania, Malawi, Mozambique and Madagascar), western (Burkina Faso, Mali, Nigeria and western Kenya), and central (Gabon, coastal Angola). A. funestus from the southwest of Uganda had affinities to all three subdivisions. Mitochondrial DNA (mtDNA) corroborated this structure, although mtDNA gene trees showed less resolution. The eastern subdivision had significantly lower diversity, similar to the pattern found in the codistributed malaria vector Anopheles gambiae. This suggests that both species have responded to common geographic and/or climatic constraints. The western division showed signatures of population expansion encompassing Kenya west of the Rift Valley through Burkina Faso and Mali. This pattern also bears similarity to A. gambiae, and may reflect a common response to expanding human populations following the development of agriculture. Due to the presumed recent population expansion, the correlation between genetic and geographic distance was weak. Mitochondrial DNA revealed further cryptic subdivision in A. funestus, not detected in the nuclear genome. Mozambique and Madagascar samples contained two mtDNA lineages, designated clade I and clade II, that were separated by two fixed differences and an average of 2% divergence, which implies that they have evolved independently for approximately 1 million years. Clade I was found in all 11 locations, whereas clade II was sampled only on Madagascar and Mozambique. We suggest that the latter clade may represent mtDNA capture by A. funestus, resulting from historical gene flow either among previously isolated and divergent populations or with a related species. PMID- 16313590 TI - Sequence diversity of the MHC DRB gene in the Eurasian beaver (Castor fiber). AB - Major histocompatibility complex (MHC) genes, coding molecules which play an important role in immune response, are the most polymorphic genes known in vertebrates. However, MHC polymorphism in some species is limited. MHC monomorphism at several MHC class I and II loci was previously reported for two neighbouring northern European populations of the Eurasian beaver (Castor fiber) and reduced selection for polymorphism has been hypothesized. Here, we analysed a partial sequence of the second exon of the MHC II DRB locus from seven relict European and Asian beaver populations. We detected 10 unique alleles among 76 beavers analysed. Only a western Siberian population was polymorphic, with four alleles detected in 10 individuals. Each of the remaining populations was fixed for a different allele. Sequences showed considerable divergence, suggesting the long persistence of allelic lineages. A significant excess of nonsynonymous substitutions was detected at the antigen binding sites, indicating that sequence evolution of beaver DRB was driven by positive selection. Current MHC monomorphism in the majority of populations may be the result of the superimposition of the recent bottleneck on pre-existing genetic structure resulting from population subdivision and differential pathogen pressure. PMID- 16313591 TI - Subterranean phylogeography of freshwater crayfishes shows extensive gene flow and surprisingly large population sizes. AB - Subterranean animals are currently viewed as highly imperiled, precariously avoiding extinction in an extreme environment of darkness. This assumption is based on a hypothesis that the reduction in visual systems and morphology common in cave faunas reflects a genetic inability to adapt and persist coupled with the perception of a habitat that is limited, disconnected, and fragile. Accordingly, 95% of cave fauna in the United States are presumed endangered due to surface environmental degradation and limited geographic distributions. Our study explores the subterranean phylogeography of stygobitic crayfishes in the southeastern United States, a global hotspot of groundwater biodiversity, using extensive geographic sampling and molecular data. Despite their endangered status, our results show that subterranean crayfish species have attained moderate to high levels of genetic diversity over their evolutionary histories with large population sizes and extensive gene flow among karst systems. We then compare the subterranean population histories to those of common surface stream dwelling crayfishes. Our results show recent drastic declines in genetic variability in the surface crayfish and suggest that these species also warrant conservation attention. PMID- 16313592 TI - High genetic diversity in French invasive populations of common ragweed, Ambrosia artemisiifolia, as a result of multiple sources of introduction. AB - Ambrosia artemisiifolia is an aggressive North American annual weed, found particularly in sunflower and corn fields. Besides its economic impact on crop yield, it represents a major health problem because of its strongly allergenic pollen. Ragweed was imported inadvertently to Europe in the 18th century and has become invasive in several countries, notably in the Rhone Valley of France. It has recently expanded in both the Provence-Alpes-Cote-d'Azur and Bourgogne regions. As first steps towards understanding the causes and mechanisms of ragweed invasion, genetic variability of French and North American populations was analysed using microsatellites. Overall genetic variability was similar in North America and in the Rhone-Alpes region, but within-population levels of genetic variability were surprisingly lower in native than in invasive French populations. French populations also exhibited lower among-population differentiation. A significant pattern of isolation by distance was detected among North American populations but not among French populations. Assignment tests and distribution of rare alleles did not point to a single origin for all French populations, nor for all individuals within populations and private alleles from different North American populations were found in the same French populations. Indeed, within all French populations, individual plants were roughly equally assigned to the different North American populations. Altogether, these results suggest that the French invasive populations include plants from a mixture of sources. Reduced diversity in populations distant from the original area of introduction indicated that ragweed range expansion probably occurred through sequential bottlenecks from the original populations, and not from subsequent new introductions. PMID- 16313593 TI - Microsatellite variation within and among populations of Oryza officinalis (Poaceae), an endangered wild rice from China. AB - Oryza officinalis Wall. ex Watt. is an agriculturally important but seriously endangered species of wild rice. To obtain more accurate estimates of population structure for improved conservation planning of the species, genetic variability at 14 microsatellite DNA loci was examined in population samples covering most of the species' range in China. Considerable genetic variability (overall Na = 1.886, P = 62%, HO = 0.056, HE = 0.216, and HS = 0.277) was detected at the 14 loci in 442 individuals of the 18 natural populations. The evaluation of partitioning of genetic variability (FST = 0.442) suggested high genetic differentiation among the Chinese O. officinalis populations. An overall value of Nm = 0.316 suggested limited gene flow occurred among the sampled populations. Most of the populations showed heterozygote deficits in tests of Hardy-Weinberg equilibrium and significantly positive FIS values. This could be due to some inbreeding occurring in this predominantly outcrossing species. For effective in situ conservation and restoration genetics, maintenance of significant historical processes is particularly important, including high outbreeding, considerable gene flow, and large population effective sizes. The high FST values detected among populations in this study are instructive for adopting a conservation plan that includes representative populations with the greatest genetic variation for either in situ conservation management or germplasm collection expeditions. PMID- 16313594 TI - Genetic population structure of Natterer's bats explained by mating at swarming sites and philopatry. AB - During autumn 'swarming', large numbers of temperate bats chase each other in and around underground sites. Swarming has been proposed to be a mating event, allowing interbreeding between bats from otherwise isolated summer colonies. We studied the population structure of the Natterer's bat (Myotis nattereri), a swarming species in northern England, by sampling bats at seven sites in two swarming areas and at 11 summer colonies. Analysis of molecular variance (amova) and genetic assignment analyses showed that the swarming areas (60 km apart) support significantly different populations. A negative correlation was found between the distance of a summer colony from a swarming area and the assignment of bats to that area. High gene diversity was found at all sites (HE = 0.79) suggesting high gene flow. This was supported by a low FST (0.017) among summer colonies and the absence of isolation by distance or substructure among colonies which visit one swarming area. The FST, although low, was significantly different from zero, which could be explained by a combination of female philopatry and male-mediated gene flow through mating at swarming sites with bats from other colonies. Modelling suggested that if effective size of the summer colonies (Ne) was low to moderate (10-30), all mating must occur at the swarming sites to account for the observed FST. If the Ne was higher (50), in addition to random mating during swarming, there may be nonrandom mating at swarming sites or some within-colony mating. Conservation of swarming sites that support potentially large populations is discussed. PMID- 16313595 TI - Direct and indirect estimates of gene flow among wild and managed populations of Polaskia chichipe, an endemic columnar cactus in Central Mexico. AB - Microsatellite markers were used to obtain direct and indirect estimates of gene flow in populations of Polaskia chichipe under different management regimes, in order to understand the genetic consequences of gene flow in the evolutionary process of domestication. P. chichipe is a columnar cactus endemic to the Tehuacan Valley, Central Mexico, and has come under domestication for its edible fruit. Morphological, phenological, physiological, and reproductive differences, apparently attributable to artificial selection, exist between wild and managed populations, which grow sympatrically. However, strong gene flow may counteract the effects of this selection. In this study, we used paternity analysis to demonstrate that although most of the pollinations occur among individuals within the same population at distances < 40 m, pollen flow from other populations is considerable (27 +/- 5%). Heterogeneity in pollen clouds sampled by mother plants (FST = 0.12) indicated nonrandom mating, which is probably due to temporal heterogeneity in pollen movement. Spatial structure on local and regional scales is consistent with an isolation-by-distance model. The similarity of indirect, direct and demographic estimates of neighbourhood size (74-250 individuals) suggests that this genetic structure is representative of an equilibrium state. These results suggest that traditional management practices have conserved the genetic resources of this species in situ, but also that gene flow is counteracting the effect of domestication to some degree. We discuss our results in the general context of genetic exchange between cultivated and wild populations during the domestication process. PMID- 16313596 TI - Pan-European phylogeography of the aquatic snail Theodoxus fluviatilis (Gastropoda: Neritidae). AB - Investigating the geographical distribution of genetic lineages within species is critical to our understanding of how species evolve. As many species inhabit large and complex ranges, it is important that phylogeographical research take into account the entire range of widespread species to clarify how myriad extrinsic variables have affected their evolutionary history. Using phylogenetic, nested clade, and mismatch distribution analyses on a portion of the mitochondrial COI gene, I demonstrate that the wide-ranging freshwater snail Theodoxus fluviatilis possesses in parallel many of the phylogeographical patterns seen in less widespread freshwater species of Europe. Fragmentary forces play a major part in structuring the range of this species, with 12 of 14 geographically structured nested clades displaying a distribution consistent with fragmentation or restricted dispersal. Certain regions of southern Europe harbour the majority of genetic diversity (total haplotype diversity, H = 0.87), particularly Italy (H = 0.87) and areas surrounding the Black Sea (H = 0.81). Post-Pleistocene range expansion is pronounced, with the majority of northern European populations (95% of sample sites) having arisen from northern Italian individuals that initially colonized northern Germany. Additionally, two highly divergent haplotype lineages present in northern Germany imply that there were at least two postglacial recolonization routes. Estuaries may also provide a means of dispersal given that no genetic differentiation was found between estuarine populations and neighbouring freshwater populations. Taken together, these data reveal a species with a complex genetic history resulting from the fragmentary effects of European geology as well as continuous and discrete range expansion related to their aquatic biology. PMID- 16313597 TI - Colour vision and speciation in Lake Victoria cichlids of the genus Pundamilia. AB - Lake Victoria cichlids are one of the most speciose groups of vertebrates. Selection on coloration is likely playing an important role in their rapid speciation. To test the hypothesis that sensory biases could explain species differences in mating preferences and nuptial coloration, we studied seven populations of four closely related species of the genus Pundamilia that differ in visual environment and male nuptial colour. Microspectrophotometry determined that the wavelength of maximum absorption (lambdamax) of the rod pigment and three cone pigments were similar in all four species. Only the long wavelength sensitive (LWS) pigment varied among species, with 3-4 nm shifts in lambdamax that correlated with differences in the LWS opsin sequence. These subtle shifts in lambdamax coincided with large shifts in male body colour, with red species having longer LWS pigments than blue species. Furthermore, we observed within and between species a correlation between water transparency and the proportion of red/red vs. red/green double cones. Individuals from turbid water had more red/red double cones than individuals from clear water. The variation in LWS lambdamax and in the proportion of red/red double cones could lead to differences in perceived brightness that may explain the evolution of variation in male coloration. However, other factors, such as chromophore shifts and higher order neural processing, should also be investigated to fully understand the physiological basis of differential responses to male mating hues in cichlid fish. PMID- 16313598 TI - The colonization of Europe by the freshwater crustacean Asellus aquaticus (Crustacea: Isopoda) proceeded from ancient refugia and was directed by habitat connectivity. AB - Recent continental-scale phylogeographic studies have demonstrated that not all freshwater fauna colonized Europe from the classic Mediterranean peninsular refugia, and that northern or central parts of the continent were occupied before, and remained inhabited throughout the Pleistocene. The colonization history of the ubiquitous aquatic isopod crustacean Asellus aquaticus was assessed using mitochondrial COI and a variable part of nuclear 28S rDNA sequences. Phylogeographic analysis of the former suggested that dispersion proceeded possibly during late Miocene from the western part of the Pannonian basin. Several areas colonized from here have served as secondary refugia and/or origins of dispersion, well before the beginning of the Pleistocene. Postglacial large-scale range expansion was coupled with numerous separate local dispersions from different refugial areas. Connectivity of the freshwater habitat has played an important role in shaping the current distribution of genetic diversity, which was highest in large rivers. The importance of hydrographic connections for the maintenance of genetic contact was underscored by a discordant pattern of mtDNA and nuclear rDNA differentiation. Individuals from all over Europe, differing in their mtDNA to a level normally found between species or even genera (maximal within population nucleotide divergence reached 0.16 +/- 0.018), shared the same 28S rRNA gene sequence. Only populations from hydrographically isolated karst water systems in the northwestern Dinaric Karst had distinct 28S sequences. Here isolation seemed to be strong enough to prevent homogenization of the rRNA gene family, whereas across the rest of Europe genetic contact was sufficient for concerted evolution to act. PMID- 16313599 TI - Evolutionary history of the European whitefish Coregonus lavaretus (L.) species complex as inferred from mtDNA phylogeography and gill-raker numbers. AB - We compared mitochondrial DNA and gill-raker number variation in populations of the European whitefish Coregonus lavaretus (L.) species complex to illuminate their evolutionary history, and discuss mechanisms behind diversification. Using single-strand conformation polymorphism (SSCP) and sequencing 528 bp of combined parts of the cytochrome oxidase b (cyt b) and NADH dehydrogenase subunit 3 (ND3) mithochondrial DNA (mtDNA) regions, we documented phylogeographic relationships among populations and phylogeny of mtDNA haplotypes. Demographic events behind geographical distribution of haplotypes were inferred using nested clade analysis (NCA) and mismatch distribution. Concordance between operational taxonomical groups, based on gill-raker numbers, and mtDNA patterns was tested. Three major mtDNA clades were resolved in Europe: a North European clade from northwest Russia to Denmark, a Siberian clade from the Arctic Sea to southwest Norway, and a South European clade from Denmark to the European Alps, reflecting occupation in different glacial refugia. Demographic events inferred from NCA were isolation by distance, range expansion, and fragmentation. Mismatch analysis suggested that clades which colonized Fennoscandia and the Alps expanded in population size 24 500-5800 years before present, with minute female effective population sizes, implying small founder populations during colonization. Gill-raker counts did not commensurate with hierarchical mtDNA clades, and poorly with haplotypes, suggesting recent origin of gill-raker variation. Whitefish designations based on gill-raker numbers were not associated with ancient clades. Lack of congruence in morphology and evolutionary lineages implies that the taxonomy of this species complex should be reconsidered. PMID- 16313600 TI - Patterns of hybridization and population genetic structure in the terrestrial orchids Liparis kumokiri and Liparis makinoana (Orchidaceae) in sympatric populations. AB - We investigated the potential for gene flow and genetic assimilation via hybridization between common and rare species of the terrestrial orchid genus Liparis, focusing specifically on sympatric and allopatric populations of the common Liparis kumokiri and the rare Liparis makinoana. We utilized analyses of genetic diversity, morphology, and the spatial distributions of individuals and genotypes to quantify the dynamics of interspecific gene flow at within- and among-population scales. High levels of allozyme genetic diversity (HE) were found in populations of the rare L. makinoana (0.317), whereas the common L. kumokiri (N = 1744 from 14 populations) revealed a complete lack of variation. This contrast may reflect different breeding systems and associated rates of genetic drift (L. makinoana is self-incompatible, whereas L. kumokiri is self compatible). At the two known sympatric sites, individuals were found that recombined parental phenotypes, possessing floral characteristics of L. kumokiri and vegetative characteristics of L. makinoana. These putative hybrids were the only individuals found segregating alleles diagnostic of both parental species. Analysis of these individuals indicated that hybrid genotypes were skewed towards L. kumokiri and later generation recombinants of L. kumokiri at both sympatric sites. Furthermore, Ripley's bivariate L(r) statistics revealed that at one site these hybrids are strongly spatially clustered with L. kumokiri. Nonetheless, the relatively low frequency of hybrids, absence of ongoing hybridization (no F1s or first generation backcrossess), and strong genetic differentiation between morphologically 'pure' parental populations at sympatric sites (FST = 0.708 0.816) indicates that hybridization was not an important bridge for gene flow. The results from these two species suggest that natural hybridization has not played an important role in the diversification of Liparis, but instead support the view that genetic drift and limited gene flow are primarily responsible for speciation in Liparis. Based on genetic data and current status of the species, implications of the research for conservation are considered to provide guidelines for appropriate conservation and management strategies. PMID- 16313601 TI - Multiple origin of metallicolous populations of the pseudometallophyte Arabidopsis halleri (Brassicaceae) in central Europe: the cpDNA testimony. AB - The population structure of the pseudo-metallophyte herb, Arabidopsis halleri, was studied using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) on chloroplast DNA (cpDNA). The history of metallicolous (M) populations showing increased zinc tolerance was investigated. Eight primer enzyme combinations out of 72 tested were applied to a total of 625 individuals from 28 widespread populations, 14 of them being M. Eleven distinct chlorotypes were found: five were common to nonmetallicolous (NM) and M populations, whereas six were only observed in one edaphic type (five in NM and one in M). No difference in chlorotype diversity between edaphic types was detected. Computed on the basis of chlorotype frequencies, the level of population differentiation was high but remained the same when taking into account levels of molecular divergence between chlorotypes. Isolation by distance was largely responsible for population differentiation. Geographically isolated groups of M populations were more genetically related to their closest NM populations than to each other. Our results suggest that M populations have been founded separately from distinct NM populations without suffering founding events and that the evolution towards increased tolerance observed in the distinct M population groups occurred independently. PMID- 16313602 TI - Refugia, dispersal and divergence in a forest archipelago: a study of Streptocarpus in eastern South Africa. AB - We describe a scenario of plant speciation across a relict forest archipelago in South Africa involving Pleistocene habitat expansion-contraction cycles, dispersal and adaptation to lower temperatures. This is the first population level study using molecular data in South African forests and has significant implications for conservation efforts in this area. Populations of the mesophytic forest floor herbs Streptocarpus primulifolius sensu lato and Streptocarpus rexii were sampled throughout their range in the naturally fragmented forests of eastern South Africa in order to investigate population genetic and phylogenetic patterns within the species complex, using nuclear microsatellites, nuclear ribosomal ITS (internal transcribed spacer) sequences and chloroplast genome sequences. S. primulifolius harbours high levels of genetic diversity at both the nuclear (mean HE = 0.50) and the chloroplast level (each population fixed for a unique haplotype). This is consistent with populations of these coastal species being Pleistocene relicts. In contrast, populations of S. rexii in cooler habitats at higher altitudes and lower latitudes harbour little or no nuclear genetic diversity (mean HE = 0.09) and most share a common chloroplast haplotype. The split of S. rexii from populations intermediate between the two species (S. cf. primulifolius) occurred between 0 and 0.44 million years ago according to the calibrated ITS phylogeny of the taxa. The low genetic diversity and homogeneity of S. rexii is congruent with this species having reached its current range during the Holocene. We found no evidence of monophyly of any of the taxa in this study, which we consider a consequence of recent evolution in a fragmented habitat. PMID- 16313603 TI - Hidden diversity and host specificity in cycliophorans: a phylogeographic analysis along the North Atlantic and Mediterranean Sea. AB - In order to elucidate the evolutionary history and the population structure of the members of the phylum Cycliophora, which live commensally on three species of lobsters, we studied sequence variation in the mitochondrial gene cyctochrome c oxidase subunit I. Overall 242 sequences from 16 locations on both coasts of the North Atlantic, including the North Sea and the Mediterranean, were analysed, revealing 28 haplotypes, with a maximum sequence divergence of 16.6%. Total genetic diversity was high (h = 0.8322, pi = 0.0898), as it was for the commensals on Homarus americanus (17 haplotypes, h = 0.7506, pi = 0.0504). However, it was low for commensals on Nephrops norvegicus (6 haplotypes, h = 0.3899, pi = 0.0035), and intermediate for cycliophorans on Homarus gammarus (5 haplotypes, h = 0.3020, pi = 0.0140). Although two of the host lobsters co inhabit the coastal waters of Europe, a strong genetic structure (78.45% of the observed genetic variation) was detected among populations on all host species, indicating the existence of a reproductively isolated species on each lobster. In addition, genetic structure over long distances exists among populations on each host species. Such patterns can be explained by the limited dispersal ability of the cycliophoran chordoid larva. Demographic and phylogenetic analyses suggest old and possibly cryptic populations present on H. americanus and H. gammarus, while the latter may have experienced recent bottlenecks, perhaps during Pleistocene glaciations. Populations on N. norvegicus appear to be of recent origin. PMID- 16313604 TI - Pollen flow in the wildservice tree, Sorbus torminalis (L.) Crantz. II. Pollen dispersal and heterogeneity in mating success inferred from parent-offspring analysis. AB - Knowing the extent of gene movements from parents to offspring is essential to understand the potential of a species to adapt rapidly to a changing environment, and to design appropriate conservation strategies. In this study, we develop a nonlinear statistical model to jointly estimate the pollen dispersal kernel and the heterogeneity in fecundity among phenotypically or environmentally defined groups of males. This model uses genotype data from a sample of fruiting plants, a sample of seeds harvested on each of these plants, and all males within a circumscribed area. We apply this model to a scattered, entomophilous woody species, Sorbus torminalis (L.) Crantz, within a natural population covering more than 470 ha. We estimate a high heterogeneity in male fecundity among ecological groups, both due to phenotype (size of the trees and flowering intensity) and landscape factors (stand density within the neighbourhood). We also show that fat tailed kernels are the most appropriate to depict the important abilities of long distance pollen dispersal for this species. Finally, our results reveal that the spatial position of a male with respect to females affects as much its mating success as ecological determinants of male fecundity. Our study thus stresses the interest to account for the dispersal kernel when estimating heterogeneity in male fecundity, and reciprocally. PMID- 16313605 TI - Microsatellite DNA analysis shows that greater sage grouse leks are not kin groups. AB - The spectacular social courtship displays of lekking birds are thought to evolve via sexual selection, but this view does not easily explain the participation of many males that apparently fail to mate. One of several proposed solutions to this 'lek skew paradox' is that kin selection favours low-ranking males joining leks to increase the fitness of closely related breeders. We investigated the potential for kin selection to operate in leks of the greater sage grouse, Centrocercus urophasianus, by estimating relatedness between lekking males using microsatellite DNA markers. We also calibrated these estimates using data from known families. Mean relatedness within leks was statistically indistinguishable from zero. We also found no evidence for local clustering of kin during lek display, although males tended to range closer to kin when off the lek. These results make kin selection an unlikely solution to the lek skew paradox in sage grouse. Together with other recent studies, they also raise the question of why kin selection apparently promotes social courtship in some lekking species, but not in others. PMID- 16313606 TI - Detection of secondary predation by PCR analyses of the gut contents of invertebrate generalist predators. AB - Predation by generalist predators is difficult to study in the field because of the complex effects of positive and negative interactions within and between predator species and guilds. Predation can be monitored by molecular means, through identification of prey DNA within predators. However, polymerase chain reaction (PCR) amplification of prey DNA from predators cannot discriminate between primary and secondary predation (hyperpredation), in which one predator feeds on another that has recently eaten the target prey. Here we quantify, for the first time, the potential error caused by detection of prey DNA following secondary predation, using an aphid-spider-carabid model. First, the aphid Sitobion avenae was fed to the spider Tenuiphantes tenuis and the carabid Pterostichus melanarius, and the postconsumption detection periods, for prey DNA within predators, were calculated. Aphids were then fed to spiders and the spiders to carabids. Aphid DNA was detected in the predators using primers that amplified 245- and 110-bp fragments of the mitochondrial cytochrome oxidase I gene. Fragment size and predator sex had no significant effect on detection periods. Secondary predation could be detected for up to 8 h, when carabids fed on spiders immediately after the latter had consumed aphids. Beetles tested positive up to 4 h after eating spiders that had digested their aphid prey for 4 h. Clearly, the extreme sensitivity of PCR makes detection of secondary predation more likely, and the only reliable answer in future may be to use PCR to identify, in parallel, instances of intraguild predation. PMID- 16313607 TI - Interannual genetic heterogeneity of pollen pools accepted by Quercus salicina individuals. AB - Since flowering often varies among years in wind-pollinated woody species, the genetic composition of pollen pools accepted by seed parents can differ between years. The interannual heterogeneity of pollen flow may be important for maintaining genetic diversity within populations because it can increase genetic variation within populations and the effective sizes of the populations. In this study we examined heterogeneity, using paternity analysis and analysis of molecular variance, in the genetic composition of pollen pools among different reproductive years for six Quercus salicina seed parents in an 11.56-ha plot in a temperate old-growth evergreen broadleaved forest. The genotypes at seven microsatellite loci were determined for 111 adult trees and 777 offspring of the six seed parents in 2-5 reproductive years. Genetic differentiation of pollen pools among different reproductive years for each seed parent was significant over all seed parents and for each of four seed parents that were analysed for more than 2 years, but not for either of the other two seed parents (analysed for 2 years). For both the pollen pools originating from inside the plot and those originating from outside it, genetic differentiation among different reproductive years for each seed parent was significant over all seed parents. However, among year genetic differentiation in the pollen pools originating from within the plot was detected for all four of the seed parents that were analysed for more than 2 years, but for only one of the four in the pools originating from outside the plot. Genetic diversity (estimated as allelic richness and gene diversity) was higher for pollen pools over all reproductive years than for pollen pools in single years. These results indicate that the year-to-year genetic variation of pollen pools increases genetic diversity in offspring and is strongly affected by the variation in pollen parents within the plot because of their high pollination contributions. The high year-to-year variation in pollen parents within the plot and overall supports the hypothesis that the offspring produced across years represent a larger genetic neighbourhood. PMID- 16313608 TI - Local forest environment largely affects below-ground growth, clonal diversity and fine-scale spatial genetic structure in the temperate deciduous forest herb Paris quadrifolia. AB - Paris quadrifolia (herb Paris) is a long-lived, clonal woodland herb that shows strong differences in local population size and shoot density along an environmental gradient of soil and light conditions. This environmentally based structuring may be mediated by differences in clonal growth and seedling recruitment through sexual reproduction. To study the interrelationship between environmental conditions and spatial patterns of clonal growth, the spatial genetic structure of four P. quadrifolia populations growing in strongly contrasting sites was determined. In the first place, plant excavations were performed in order to (i) determine differences in below-ground growth of genets, (ii) investigate connectedness of ramets and (iii) determine total genet size. Although no differences in internode length were found among sites, clones in moist sites were much smaller (genets usually consisted of 1-3 interconnected shoots, most of them flowering) than genets in dry sites, which consisted of up to 15 interconnected shoots, the majority of which were vegetative. Further, amplified fragment length polymorphism (AFLP) markers were used. Clonal diversity was higher in populations located in moist and productive ash-poplar forests compared to those found in drier and less productive mixed forest sites (G/N: 0.27 and 0.14 and Simpson's D: 0.84 and 0.75, respectively). Patterns of spatial population genetic structure under dry conditions revealed several large clones dominating the entire population, whereas in moist sites many small genets were observed. Nevertheless, strong spatial genetic structure of the genet population was observed. Our results clearly demonstrate that patterns of clonal diversity and growth form of P. quadrifolia differ among environments. Limited seedling recruitment and large clone sizes due to higher connectedness of ramets explain the low clonal diversity in dry sites. In moist sites, higher levels of clonal diversity and small clone sizes indicate repeated seedling recruitment, whereas strong spatial genetic structure suggests limited seed dispersal within populations. PMID- 16313609 TI - Bacterial metal detectors. AB - Gram negative bacteria can detect environmental iron using outer membrane transporters (OMTs), and then regulate certain transport genes to take advantage of a readily available iron source. This process begins with an iron complex being bound by an OMT, and results in a signal being sent across the outer membrane, the periplasmic space, and the inner membrane, to a sigma factor that interacts with RNA polymerase and initiates transcription of relevant genes. Many of the interactions contributing to signalling have been observed by genetic and biochemical studies, but structural studies, which potentially show these interactions in molecular detail, have been limited. In this issue, Garcia Herrero and Vogel describe an NMR structure of the periplasmic domain of an OMT, which had not been seen in previous X-ray crystal structures. This domain transmits the 'iron availability' signal to the next protein in the signal transduction cascade, which sits in the inner membrane and extends into the periplasm. The new structure extends our knowledge of transporter architecture and suggests how signalling may occur across the outer membrane. PMID- 16313610 TI - Iron-reducing bacteria unravel novel strategies for the anaerobic catabolism of aromatic compounds. AB - Although the aerobic degradation of aromatic compounds has been extensively studied in many microorganisms, the anaerobic mineralization of the aromatic ring is a more recently discovered microbial capacity on which very little information is available from facultative anaerobic bacteria. In this issue of Molecular Microbiology, Wischgoll and colleagues use proteomic and reverse-transcription polymerase chain reaction (PCR) approaches to identify for the first time the gene clusters involved in the central pathway for the catabolism of aromatic compounds in Geobacter metallireducens, a strictly anaerobic iron-reducing bacterium. This work highlights that the major difference in anaerobic benzoate metabolism of facultative and strictly anaerobic bacteria is the reductive process for dearomatization of benzoyl-CoA. The authors propose that a new type of benzoyl-CoA reductase, comprising molybdenum- and selenocysteine-containing proteins, is present in strictly anaerobic bacteria. This work paves the way to fundamental studies on the biochemistry and regulation of this new reductive process and provides the first genetic clues on the anaerobic catabolism of benzoate by strict anaerobes. PMID- 16313611 TI - Molecular architecture and function of the Omp85 family of proteins. AB - Omp85 is a protein found in Gram-negative bacteria where it serves to integrate proteins into the bacterial outer membrane. Members of the Omp85 family of proteins are defined by the presence of two domains: an N-terminal, periplasmic domain rich in POTRA repeats and a C-terminal beta-barrel domain embedded in the outer membrane. The widespread distribution of Omp85 family members together with their fundamental role in outer membrane assembly suggests the ancestral Omp85 arose early in the evolution of prokaryotic cells. Mitochondria, derived from an ancestral bacterial endosymbiont, also use a member of the Omp85 family to assemble proteins in their outer membranes. More distant relationships are seen between the Omp85 family and both the core proteins in two-partner secretion systems and the Toc75 family of protein translocases found in plastid outer envelopes. Aspects of the ancestry and molecular architecture of the Omp85 family of proteins is providing insight into the mechanism by which proteins might be integrated and assembled into bacterial outer membranes. PMID- 16313612 TI - Nuclear magnetic resonance solution structure of the periplasmic signalling domain of the TonB-dependent outer membrane transporter FecA from Escherichia coli. AB - Gram-negative bacteria possess outer membrane receptors that utilize energy provided by the TonB system to take up iron. Several of these receptors participate in extracytoplasmic factor (ECF) signalling through an N-terminal signalling domain that interacts with a periplasmic transmembrane anti-sigma factor protein and a cytoplasmic sigma factor protein. The structures of the intact TonB-dependent outer membrane receptor FecA from Escherichia coli and FpvA from Pseudomonas aeruginosa have recently been solved by protein crystallography; however, no electron density was detected for their periplasmic signalling domains, suggesting that it was either unfolded or flexible with respect to the remainder of the protein. Here we describe the well-defined solution structure of this domain solved by multidimensional nuclear magnetic resonance (NMR) spectroscopy. The monomeric protein construct contains the 79-residue N-terminal domain as well as the next 17 residues that are part of the receptor's plug domain. These form two clearly distinct regions: a highly structured domain at the N-terminal end followed by an extended flexible tail at the C-terminal end, which includes the 'TonB-box' region, and connects it to the plug domain of the receptor. The structured region consists of two alpha-helices that are positioned side by side and are sandwiched in between two small beta-sheets. This is a novel protein fold which appears to be preserved in all the periplasmic signalling domains of bacterial TonB-dependent outer membrane receptors that are involved in ECF signalling, because the hydrophobic residues that make up the core of the protein domain are highly conserved. PMID- 16313613 TI - Gene clusters involved in anaerobic benzoate degradation of Geobacter metallireducens. AB - The degradation of aromatic compounds follows different biochemical principles in aerobic and anaerobic microorganisms. While aerobes dearomatize and cleave the aromatic ring by oxygenases, facultative anaerobes utilize an ATP-dependent ring reductase for the dearomatization of the activated key intermediate benzoyl coenzyme A (CoA). In this work, the aromatic metabolism was studied in the obligately anaerobic model organism Geobacter metallireducens. The gene coding for a putative carboxylic acid-CoA ligase was heterologously overexpressed and the gene product was characterized as a highly specific benzoate-CoA ligase catalysing the initial step of benzoate metabolism. However, no evidence for the presence of an ATP-dependent benzoyl-CoA reductase as observed in facultative anaerobes was obtained. In a proteomic approach benzoate-induced proteins were identified; the corresponding genes are organized in two clusters comprising 44 genes. Induction of representative genes during growth on benzoate was confirmed by reverse transcription polymerase chain reaction. The results obtained suggest that benzoate is activated to benzoyl-CoA, which is then reductively dearomatized to cyclohexa-1,5-diene-1-carbonyl-CoA, followed by beta-oxidation reactions to acetyl-CoA units, as in facultatively anaerobic bacteria. However, in G. metallireducens the process of reductive benzene ring dearomatization appears to be catalysed by a set of completely different protein components comprising putative molybdenum and selenocysteine containing enzymes. PMID- 16313614 TI - An atypical mitogen-activated protein kinase controls cytokinesis and flagellar motility during male gamete formation in a malaria parasite. AB - The transmission of malaria parasites to the mosquito depends critically on the rapid initiation of sexual reproduction in response to triggers from the mosquito midgut environment. We here identify an essential function for an atypical mitogen-activated protein kinase of the rodent malaria parasite Plasmodium berghei, Pbmap-2, in male sexual differentiation and parasite transmission to the mosquito. A deletion mutant no longer expressing the Pbmap-2 protein develops as wild type throughout the asexual erythrocytic phase of the life cycle. Gametocytes, the sexual transmission stages, form normally and respond in vitro to the appropriate environmental cues by rounding up and emerging from their host cells. However, microgametocytes fail to release flagellated microgametes. Female development is not affected, as judged by the ability of macrogametes to become cross-fertilized by microgametes from a donor strain. Cellular differentiation of Pbmap-2 KO microgametocytes is blocked at a late stage of male gamete formation, after replication and mitoses have been completed and axonemes have been assembled. These data demonstrate a function for Pbmap-2 in initiating cytokinesis and axoneme motility, possibly downstream of a cell cycle checkpoint for the completion of replication and/or mitosis, which are extraordinarily rapid in the male gametocyte. PMID- 16313615 TI - Two proteins with 6-cys motifs are required for malarial parasites to commit to infection of the hepatocyte. AB - Many intracellular pathogens have host cells suitable for their proliferation, and selectively invade them using specific host-parasite interactions. Malarial sporozoites, the liver-invasive forms, are effectively targeted to hepatocytes and proliferate in them. So far, however, sporozoite molecules that mediate the specific infection of hepatocytes remain unknown. Here we report that two proteins, Pbs36p and Pbs36, belonging to the plasmodium 6-cys domain protein family, carry out this function. We found that these molecules are specifically produced in liver-infective sporozoites. Target disruption of the respective genes nearly abolished sporozoite infectivity in the mammalian host. Invasion assays revealed that the mutant parasites could not commit to infection, even when they encounter with hepatocytes, resulting in continuous traversal of hepatocytes. These results suggest that these proteins are necessary for sporozoites to recognize hepatocytes and commit to infection. This finding might lead to novel anti-malarial strategies that prevent sporozoite infection of the hepatocyte. PMID- 16313616 TI - Cross-regulation among disparate antibiotic biosynthetic pathways of Streptomyces coelicolor. AB - A complex programme of regulation governs gene expression during development of the morphologically and biochemically complex eubacterial genus Streptomyces. Earlier work has suggested a model in which 'higher level' pleiotropic regulators activate 'pathway-specific' regulators located within chromosomal gene clusters encoding biosynthesis of individual antibiotics. We used mutational analysis and adventitious overexpression of key Streptomyces coelicolor regulators to investigate functional interactions among them. We report here that cluster situated regulators (CSRs) thought to be pathway-specific can also control other antibiotic biosynthetic gene clusters, and thus have pleiotropic actions. Surprisingly, we also find that CSRs exhibit growth-phase-dependent control over afsR2/afsS, a 'higher level' pleiotropic regulatory locus not located within any of the chromosomal gene clusters it targets, and further demonstrate that cross regulation by CSRs is modulated globally and differentially during the S. coelicolor growth cycle by the RNaseIII homologue AbsB. Our results, which reveal a network of functional interactions among regulators that govern production of antibiotics and other secondary metabolites in S. coelicolor, suggest that revision of the currently prevalent view of higher-level versus pathway-specific regulation of secondary metabolism in Streptomyces species is warranted. PMID- 16313617 TI - Candida albicans MTLalpha tup1Delta mutants can reversibly switch to mating competent, filamentous growth forms. AB - Candida albicans strains that are homozygous at the mating type locus (MTLa or MTLalpha) can spontaneously switch from the normal round-to-oval yeast cell morphology to an elongated, so-called opaque cell form that can mate with opaque cells of the opposite mating type. In response to environmental signals, C. albicans also undergoes a transition from yeast to filamentous growth, which is negatively regulated by the general repressor Tup1p. Therefore, C. albicans mutants in which the TUP1 gene is inactivated grow constitutively in the filamentous form. We found that tup1Delta mutants of the MTLalpha strain WO-1 are still able to undergo phenotypic switching. Although the mutants had lost the capacity to grow in the normal yeast (white) or opaque forms, they could still reversibly switch between four different cell and colony phenotypes (designated as fuzzy, frizzy, wrinkled and smooth) at a frequency of about 10(-3) to 10(-4). Deletion of TUP1 resulted in deregulated expression of phase-specific genes. While the white-specific WH11 gene was constitutively expressed in all four cell types, the opaque-specific SAP1 gene remained repressed and the opaque-specific OP4 gene was weakly induced in all phase variants. In spite of the loss of white- and opaque-specific cell morphology and gene expression, the tup1Delta mutants retained an important characteristic of their wild-type parent, the ability to switch to a mating-competent form. The three filamentous phase variants (fuzzy, frizzy and wrinkled) all were able to mate and produce recombinant progeny with opaque cells of an MTLa strain at frequencies that were somewhat lower than those of normal opaque cells, whereas the smooth phase variant was unable to do so. Therefore, although deletion of TUP1 in C. albicans MTLalpha cells affects cellular morphology and gene expression patterns, the mutants can still reversibly switch between mating-competent and -incompetent cell types and mate with a partner of the opposite mating type. PMID- 16313618 TI - Characterization of point mutations in the cdtA gene of the cytolethal distending toxin of Actinobacillus actinomycetemcomitans. AB - The Cdt is a family of gram-negative bacterial toxins that typically arrest eukaryotic cells in the G0/G1 or G2/M phase of the cell cycle. The toxin is a heterotrimer composed of the cdtA, cdtB and cdtC gene products. Although it has been shown that the CdtA protein subunit binds to cells in culture and in an enzyme-linked immunosorbent assay (CELISA) the precise mechanisms by which CdtA interacts with CdtB and CdtC has not yet been clarified. In this study we employed a random mutagenesis strategy to construct a library of point mutations in cdtA to assess the contribution of individual amino acids to binding activity and to the ability of the subunit to form biologically active holotoxin. Single unique amino acid substitutions in seven CdtA mutants resulted in reduced binding of the purified recombinant protein to Chinese hamster ovary cells and loss of binding to the fucose-containing glycoprotein, thyroglobulin. These mutations clustered at the 5'- and 3'-ends of the cdtA gene resulting in amino acid substitutions that resided outside of the aromatic patch region and a conserved region in CdtA homologues. Three of the amino acid substitutions, at positions S165N (mutA81), T41A (mutA121) and C178W (mutA221) resulted in gene products that formed holotoxin complexes that exhibited a 60% reduction (mutA81) or loss (mutA121, mutA221) of proliferation inhibition. A similar pattern was observed when these mutant holotoxins were tested for their ability to induce cell cycle arrest and to convert supercoiled DNA to relaxed and linear forms in vitro. The mutations in mutA81 and mutA221 disrupted holotoxin formation. The positions of the amino acid substitutions were mapped in the Haemophilus ducreyi Cdt crystal structure providing some insight into structure and function. PMID- 16313619 TI - BapA, a large secreted protein required for biofilm formation and host colonization of Salmonella enterica serovar Enteritidis. AB - In environmental settings, biofilms represent the common way of life of microorganisms. Salmonella enterica serovar Enteritidis, the most frequent cause of gastroenteritis in developed countries, produces a biofilm whose matrix is mainly composed of curli fimbriae and cellulose. In contrast to other bacterial biofilms, no proteinaceous compound has been reported to participate in the formation of this matrix. Here, we report the discovery of BapA, a large cell surface protein required for biofilm formation by S. Enteritidis. Deletion of bapA caused the loss of the capacity to form a biofilm whereas the overexpression of a chromosomal copy of bapA increased the biofilm biomass formation. We provide evidence that overproduction of curli fimbriae and not cellulose can compensate for the biofilm deficiency of a bapA mutant strain. BapA is secreted through a type I protein secretion system (BapBCD) situated downstream of the bapA gene and was found to be loosely associated with the cell surface. Experiments with mixed bacterial populations positive or negative for BapA showed that BapA minus cells are not recruited into the biofilm matrix. The expression of bapA is coordinated with that of genes encoding curli fimbriae and cellulose, through the action of csgD. Studies on the contribution of BapA to S. Enteritidis pathogenesis revealed that orally inoculated animals with a bapA-deficient strain survived longer than those inoculated with the wild-type strain. Also, a bapA mutant strain showed a significantly lower colonization rate at the intestinal cell barrier and consequently a decreased efficiency for organ invasion compared with the wild type strain. Taken together, these data demonstrate that BapA contributes both to biofilm formation and invasion through the regular Salmonella infection route. PMID- 16313620 TI - DnaA coordinates replication initiation and cell cycle transcription in Caulobacter crescentus. AB - The level of DnaA, a key bacterial DNA replication initiation factor, increases during the Caulobacter swarmer-to-stalked transition just before the G1/S transition. We show that DnaA coordinates DNA replication initiation with cell cycle progression by acting as a global transcription factor. Using DnaA depletion and induction in synchronized cell populations, we have analysed global transcription patterns to identify the differential regulation of normally co expressed genes. The DnaA regulon includes genes encoding several replisome components, the GcrA global cell cycle regulator, the PodJ polar localization protein, the FtsZ cell division protein, and nucleotide biosynthesis enzymes. In cells depleted of DnaA, the G1/S transition is temporally separated from the swarmer-to-stalked cell differentiation, which is normally coincident. In the absence of DnaA, the CtrA master regulator is cleared by proteolysis during the swarmer-to-stalked cell transition as usual, but DNA replication initiation is blocked. In this case, expression of gcrA, which is directly repressed by CtrA, does not increase in conjunction with the disappearance of CtrA until DnaA is subsequently induced, showing that gcrA expression requires DnaA. DnaA boxes are present upstream of many genes whose expression requires DnaA, and His6-DnaA binds to the promoters of gcrA, ftsZ and podJ in vitro. This redundant control of gcrA transcription by DnaA (activation) and CtrA (repression) forms a robust switch controlling the decision to proceed through the cell cycle or to remain in the G1 stage. PMID- 16313621 TI - Fur regulates acid resistance in Shigella flexneri via RyhB and ydeP. AB - Shigella flexneri requires iron for survival, and the genes for iron uptake and homeostasis are regulated by the Fur protein. Microarrays were used to identify genes regulated by Fur and to study the physiological effects of iron availability in S. flexneri. These assays showed that the expression of genes involved in iron acquisition and acid response was induced by low-iron availability and by inactivation of fur. A fur null mutant was acid sensitive in media at pH 2.5, and acid sensitivity was also observed in the wild-type strain grown under iron-limiting conditions. Acid resistance of the fur mutant in minimal medium was restored by addition of glutamate during acid challenge, indicating that the glutamate-dependent acid resistance system was not defective. Inactivation of ryhB, a small regulatory RNA whose expression is repressed by Fur, restored acid resistance in the fur mutant, while overexpressing ryhB increased acid sensitivity in the wild-type strain. RyhB-regulated genes were identified by microarray analysis. The expression of one of the RyhB-repressed genes, ydeP, which encodes a putative oxidoreductase, suppressed acid sensitivity in the fur mutant. Furthermore, an S. flexneri ydeP mutant was defective for both glutamate-independent and glutamate-dependent acid resistance. The repression of ydeP by RyhB may be indirect, as real time polymerase chain reaction (PCR) experiments indicated that RyhB negatively regulates evgA, which encodes an activator of ydeP. These results demonstrate that the acid sensitivity defect of the S. flexneri fur mutant is due to repression of ydeP by RyhB, most likely via repression of evgA. PMID- 16313622 TI - SapT, a lanthionine-containing peptide involved in aerial hyphae formation in the streptomycetes. AB - The developmentally complex soil microbe Streptomyces tendae secretes a hydrophobic peptide that restored to developmental mutants of S. coelicolor the ability to raise aerial hyphae. The S. tendae peptide, SapT, has a lantibiotic structure and molecular modelling predicts that it is amphiphilic, making it structurally and functionally similar to the SapB peptide produced by S. coelicolor. However, SapT, which bears three beta-methyl lanthionine bridges and one lanthionine bridge and demonstrated limited antibiotic activity, is distinct from SapB. The amphiphilic nature of both SapT and SapB is required for their ability to serve as biosurfactants facilitating the emergence of newly formed aerial hyphae. Remarkably, SapB and SapT, and the fungal hydrophobin SC3 were shown to restore to a SapB-deficient S. coelicolor mutant the capacity to undergo complete morphogenesis, such that the extracellular addition of protein resulted in sporulation. This suggests that the initiation of aerial growth may also indirectly trigger the signal transduction events needed for differentiation. These data imply that the production of morphogenetic peptides may be common among the streptomycetes, but that while their ability to function as biosurfactants is conserved, their specific lantibiotic structure is not. Finally, the identification of a second lanthionine-containing morphogenetic peptide suggests that lantibiotic structure and function may be more diverse than previously thought. PMID- 16313623 TI - Mrr instigates the SOS response after high pressure stress in Escherichia coli. AB - The bacterial SOS response is not only a vital reply to DNA damage but also constitutes an essential mechanism for the generation of genetic variability that in turn fuels adaptation and resistance development in bacterial populations. Despite the extensive depiction of the SOS regulon itself, its activation by stresses different from typical DNA damaging treatments remains poorly characterized. Recently, we reported the RecA- and LexA-dependent induction of the SOS response in Escherichia coli MG1655 after exposure to high hydrostatic pressure (HP, approximately 100 MPa), a physical stress of which the cellular effects are not well known. We now found this HP mediated SOS response to depend on RecB and not on RecF, which is a strong indication for the involvement of double strand breaks. As the pressures used in this work are thermodynamically unable to break covalent bonds in DNA, we hypothesized the involvement of a cellular function or pathway in the formation of this lesion. A specialized screening allowed us to identify the cryptic type IV restriction endonuclease Mrr as the final effector of this pathway. The HP SOS response and its corresponding phenotypes could be entirely attributed to the HP triggered activation of Mrr restriction activity. Several spontaneously occurring alleles of mrr, incapable of triggering the HP-induced SOS response, were isolated and characterized. These results provide evidence for a specific pathway that transmits the perception of HP stress to induction of the SOS response and support a role for Mrr in bacterial stress physiology. PMID- 16313624 TI - An atypical type II topoisomerase from Mycobacterium smegmatis with positive supercoiling activity. AB - Topoisomerases are essential ubiquitous enzymes, falling into two distinct classes. A number of eubacteria including Escherichia coli, typically contain four topoisomerases, two type I topoisomerases and two type II topoisomerases viz. DNA gyrase and topoisomerase IV. In contrast several other bacterial genomes including mycobacteria, encode for one type I topoisomerase and a DNA gyrase. Here we describe a new type II topoisomerase from Mycobacterium smegmatis which is different from DNA gyrase or topoisomerase IV in its characteristics and origin. The topoisomerase is distinct with respect to domain organization, properties and drug sensitivity. The enzyme catalyses relaxation of negatively supercoiled DNA in an ATP-dependent manner and also introduces positive supercoils to both relaxed and negatively supercoiled substrates. The genes for this additional topoisomerase are not found in other sequenced mycobacterial genomes and may represent a distant lineage. PMID- 16313625 TI - Ca2+-dependent lipid binding and membrane integration of PopA, a harpin-like elicitor of the hypersensitive response in tobacco. AB - PopA is released by type III secretion from the bacterial plant pathogen Ralstonia solanacearum and triggers the hypersensitive response (HR) in tobacco. The function of PopA remains obscure, mainly because mutants lacking this protein are not altered in their ability to interact with plants. In an attempt to identify the site of PopA activity in plant cells, we generated transgenic tobacco plants expressing the popA gene under the control of an inducible promoter. Immunocytologic analysis revealed that the HR phenotype of these plants correlated with the presence of PopA at the plant plasma membrane. Membrane localization was observed irrespective of whether the protein was designed to accumulate in the cytoplasm or to be secreted by the plant cell, suggesting a general lipid-binding ability. We found that the protein had a high affinity for sterols and sphingolipids in vitro and that it required Ca2+ for both lipid binding and oligomerization. In addition, the protein was integrated into liposomes and membranes from Xenopus laevis oocytes where it formed ion conducting pores. These characteristics suggest that PopA is part of a system that aims to attach the host cell plasma membrane and to allow molecules cross this barrier. PMID- 16313626 TI - Regulation of ompA mRNA stability: the role of a small regulatory RNA in growth phase-dependent control. AB - The Escherichia coli ompA mRNA, encoding a highly abundant outer membrane protein, has served as a model for regulated mRNA decay in bacteria. The half life of this transcript correlates inversely with the bacterial growth rate and is growth stage-dependent. The stability of the messenger is determined by the 5' untranslated region which possesses cleavage sites for RNase E. Hfq binds to this region, is essential for controlling the stability and has been suggested to directly regulate ompA mRNA decay. Here we report that the 78 nucleotide SraD RNA, which is highly conserved among Enterobacteriaceae, acts in destabilizing the ompA transcript when rapidly grown cells enter the stationary phase of growth. During this growth-stage the expression of SraD RNA becomes strongly increased. The SraD-mediated decay of ompA mRNA depends on Hfq and in vitro studies revealed that Hfq facilitates binding of the regulatory RNA to the translational initiation region of the messenger. Deletion of sraD, however, does not significantly affect the stability of the ompA mRNA in slowly growing cells. Our results indicate that distinct regulatory circuits are responsible for growth phase- and growth rate-dependent control of the ompA mRNA stability. PMID- 16313627 TI - A proteomic analysis of penicillin resistance in Streptococcus pneumoniae reveals a novel role for PstS, a subunit of the phosphate ABC transporter. AB - Resistance to penicillin is widespread in the Gram-positive bacterium Streptococcus pneumoniae, and while several mutations are known to be implicated in resistance other mechanisms are likely to occur. We used a proteomic screen of two independent mutants in which resistance was selected in vitro. We found a number of differentially expressed proteins including PstS, a subunit of the phosphate ABC transporter of S. pneumoniae. This protein was increased in both mutants, a phenotype correlated to increased RNA expression of the entire phosphate ABC transporter operon. Inactivation of the pstS gene led to increased susceptibility to penicillin in the wild-type strain. To further link the expression of the ABC phosphate transporter with penicillin resistance, we looked at pstS mRNA levels in 12 independent clinical isolates sensitive and resistant to penicillin and found an excellent correlation between resistance and increased expression of pstS. Inactivation of pstS in one of the clinical isolates significantly reduced penicillin resistance. Global approaches are ideally suited for the discovery of novel factors in the biology of resistance. PMID- 16313628 TI - Evidence for novel processing of the anaerobically inducible dicistronic focA-pfl mRNA transcript in Escherichia coli. AB - The anaerobically inducible dicistronic focA-pfl operon is transcribed from three co-ordinately regulated promoters that are located 5' of the operon. Remarkably, the 5' ends of four further highly abundant operon-internal transcripts are located within the focA gene, with a fifth transcript mapping in the intergenic region between focA and pfl. The findings of this study demonstrate that the bulk of these five operon-internal transcripts are the result of processing. Processing was independent of the broad-spectrum endoribonucleases associated with mRNA turnover and still occurred when the upstream regulatory region of the operon was replaced with two different heterologous promoters recognized by Escherichia coli core RNA polymerase, including the tetP promoter. However, when the T7Phi10 promoter was introduced upstream of the focA-pfl operon, mainly full length transcript and a minor amount of two processing products were observed. T7 RNA polymerase mutants that exhibit reduced elongation speed did not restore the wild-type transcript-processing pattern. Moreover, processing was independent of focA translation. Taken together, these data suggest that processing of the focA pfl transcripts occurs by a novel mechanism that might require the action of E. coli core RNA polymerase. PMID- 16313629 TI - Identification of novel Yap1p and Skn7p binding sites involved in the oxidative stress response of Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae Yap1p and Skn7p transcription factors collaborate in the activation of oxidative stress response (OSR) genes. Although Yap1p and Skn7p oxidative stress response elements (YRE, OSRE) have been characterized and identified in some OSR genes, many OSR genes lack such elements. In this study, the complex, oxidative responsive, CCP1 promoter was used as a model to investigate the cis-acting elements responsible for activation by oxidative stress. In addition to consensus YRE and OSRE sequences, novel Yap1p and Skn7p binding sites were identified in the CCP1 promoter. These new sites were found to mediate Yap1p- and Skn7p-dependent activation of OSR genes including TSA1 and CTT1 previously thought to lack Yap1p and Skn7p binding sites. The novel YREs and OSREs were found to be enriched in the promoter regions of a set of 179 OSR genes. The widespread existence of novel Yap1p and Skn7p binding sites strongly suggest that direct binding of Yap1p and Skn7p is responsible for activation of many more OSR genes than previously believed. PMID- 16313630 TI - Determinants of the streptococcal surface glycoprotein GspB that facilitate export by the accessory Sec system. AB - GspB is a large cell-surface glycoprotein expressed by Streptococcus gordonii M99 that mediates binding of this organism to human platelets. This adhesin is glycosylated in the cytoplasm, and is then transported to the cell surface via an accessory Sec system. To assess the structural features of GspB that are needed for export, we examined the effects of altering the carbohydrate moieties or the polypeptide backbone of GspB. Truncated, glycosylated variants of GspB were exported exclusively via the accessory Sec pathway. When glycosylation was abolished, the GspB variants were still exported by this pathway, but minor amounts could also be transported by the canonical Sec system. GspB variants with in-frame insertions or deletions in the N-terminus were not secreted, indicating that this domain is necessary for export. However, the N-terminus is not sufficient for the transport of heterologous proteins, because C-terminal fusion of passenger proteins to this domain hindered export. In contrast, fusion of GspB to a canonical signal peptide resulted in the efficient export of non glycosylated forms of the fusion protein via the canonical Sec pathway, whereas glycosylated forms could not be exported. Thus, the carbohydrate moieties and the atypical signal sequence of GspB interfere with export via the canonical pathway, and direct GspB towards the accessory Sec system. PMID- 16313631 TI - The cdc25 phosphatase is essential for the G2/M phase transition in the basidiomycete yeast Ustilago maydis. AB - Cdc25-related phosphatases reverse the inhibitory phosphorylation of mitotic Cyclin-dependent kinases mediated by Wee1-related kinases, thereby promoting entry into mitosis. In the fission yeast, Schizosaccharomyces pombe, Cdc25 is required for entry into mitosis, while in the budding yeast Saccharomyces cerevisiae, Mih1 (the homologue of Cdc25) is not required for entry into mitosis or for viability. As these differences were linked to the different cell division and growth mechanism of these species, we sought to analyse the roles of Cdc25 in Ustilago maydis, which as S. cerevisiae divides by budding, but relies in a polar growth. This basidiomycete yeast is perfectly suited to analyse the relationships between cell cycle and morphogenesis. We show that U. maydis contains a single Cdc25-related protein, which is essential for growth. Loss of Cdc25 function results in a specific G2 arrest that correlated with high level of Tyr15 phosphorylation of Cdk1. Moreover, we show genetic interactions of cdc25 with wee1 and clb2 that support the notion that in U. maydis Cdc25 counteracts the Wee1-mediated inhibitory phosphorylation of Cdk1-Clb2 complex. Our results supports a model in which inhibitory phosphorylation of Cdk1 is a primary mechanism operating at G2/M transition in this fungus. PMID- 16313632 TI - Plant formins come of age: something special about cross-walls. PMID- 16313633 TI - Arsenic metabolism in plants: an inside story. PMID- 16313634 TI - Phenotypic plasticity and a functional vs genetic perspective of plant gender. PMID- 16313635 TI - DNA repair mechanisms in plants: crucial sensors and effectors for the maintenance of genome integrity. AB - As obligate phototrophs, plants harness energy from sunlight to split water, producing oxygen and reducing power. This lifestyle exposes plants to particularly high levels of genotoxic stress that threatens genomic integrity, leading to mutation, developmental arrest and cell death. Plants, which with algae are the only photosynthetic eukaryotes, have evolved very effective pathways for DNA damage signalling and repair, and this review summarises our current understanding of these processes in the responses of plants to genotoxic stress. We also identify how the use of new and emerging technologies can complement established physiological and ecological studies to progress the application of this knowledge in biotechnology. PMID- 16313636 TI - Arabidopsis group Ie formins localize to specific cell membrane domains, interact with actin-binding proteins and cause defects in cell expansion upon aberrant expression. AB - The closely related proteins AtFH4 and AtFH8 represent the group Ie clade of Arabidopsis formin homologues. The subcellular localization of these proteins and their ability to affect the actin cytoskeleton were examined. AtFH4 protein activity was identified using fluorimetric techniques. Interactions between Arabidopsis profilin isoforms and AtFH4 were assayed in vitro and in vivo using pull-down assays and yeast-2-hybrid. The subcellular localization of group Ie formins was observed with indirect immunofluorescence (AtFH4) and an ethanol inducible green fluorescent protein (GFP) fusion construct (AtFH8). AtFH4 protein affected actin dynamics in vitro, and yeast-2-hybrid assays suggested isoform specific interactions with the actin-binding protein profilin in vivo. Indirect immunofluorescence showed that AtFH4 localized specifically to the cell membrane at borders between adjoining cells. Expression of an AtFH8 fusion protein resulted in GFP localization to cell membrane zones, similar to AtFH4. Furthermore, aberrant expression of AtFH8 resulted in the inhibition of root hair elongation. Taken together, these data suggest that the group Ie formins act with profilin to regulate actin polymerization at specific sites associated with the cell membrane. PMID- 16313637 TI - Regulation of cell length in the Arabidopsis thaliana root by the ethylene precursor 1-aminocyclopropane- 1-carboxylic acid: a matter of apoplastic reactions. AB - Treatment of the Arabidopsis thaliana root with the ethylene precursor 1 aminocyclopropane-1-carboxylic acid (ACC) immediately imposes a reduced maximal cell length beyond which further elongation is blocked. Here, we investigated possible apoplastic reactions involved in the inhibition of cell elongation. Five day-old Arabidopsis seedlings were transferred to a growth medium supplemented with ACC and the effect on root cell length was recorded after 3 h of treatment. Altered characteristics in the apoplast of the nonelongating cells in the ACC treated root, such as 'reactive oxygen species' (ROS) production and callose deposition, were detected using specific fluorochromes. The presence of functional hydroxyproline-rich glycoproteins (HRGPs) and the crosslinking of these cell-wall proteins are essential in limiting cell elongation. The ROS that drive the oxidative crosslinking of HRGPs, accumulate in the apoplast of cells in the zone where cell elongation stops. In the same cells, callose is deposited in the cell wall. The final cell length in the Arabidopsis root treated for a short period with ACC is determined in the zone of fast elongation. Both HRGPs crosslinking by ROS and callose deposition in the cell wall of this zone are suggested as causes for the reduced cell elongation. PMID- 16313638 TI - Uptake, translocation and transformation of arsenate and arsenite in sunflower (Helianthus annuus): formation of arsenic-phytochelatin complexes during exposure to high arsenic concentrations. AB - The aim of the study was to determine the time-dependent formation of arsenic phytochelatin (As-PC) complexes in the roots, stems and leaves of an arsenic nontolerant plant (Helianthus annuus) during exposure to 66 mol l(-1) arsenite (As(III)) or arsenate (As(V)). We used our previously developed method of simultaneous element-specific (inductively coupled plasma mass spectrometry, ICP MS) and molecular-specific (electrospray-ionization mass spectrometry, ES-MS) detection systems interfaced with a suitable chromatographic column and eluent conditions, which enabled us to identify and quantify As-PC complexes directly. Roots of As-exposed H. annuus contained up to 14 different arsenic species, including the complex of arsenite with two (gamma-Glu-Cys)(2)-Gly molecules [As((III))-(PC(2))(2)], the newly identified monomethylarsonic phytochelatin-2 or (gamma-Glu-Cys)(2)-Gly CH(3)As (MA((III))-PC(2)) and at least eight not yet identified species. The complex of arsenite with (gamma-Glu-Cys)(3)-Gly (As((III))-PC(3)) and the complex of arsenite with glutathione (GSH) and (gamma Glu-Cys)(2)-Gly (GS-As((III))-PC(2)) were present in all samples (roots, stems and leaves) taken from plants exposed to As. The GS-As((III))-PC(2) complex was the dominant complex after 1 h of exposure. As((III))-PC(3) became the predominant As-PC complex after 3 h, binding up to 40% of the As present in the exposed plants. No As-PC complexes were found in sap (mainly xylem sap from the root system), in contrast to roots, stems and leaves, which is unequivocal evidence that As-PC complexes are not involved in the translocation of As from root to leaves of H. annuus. PMID- 16313639 TI - Low frequency of transgene flow from Bt/CpTI rice to its non-transgenic counterparts planted at close spacing. AB - Crop-to-crop transgene flow will affect seed purity of non-GM rice varieties, leading to unwanted consequences. To assess the maximum probability of transgene outflow in rice (Oryza sativa), gene flow experiments were conducted with three cultivation patterns with different mixed-planting proportions of adjacent GM and non-GM rice at two sites in Fujian and Hainan Provinces of China. Three GM rice lines containing two insect-resistance genes (Bt/CpTI) and their non-GM counterparts were used in the experiments to allow natural hybridization to occur. A hygromycin resistance gene was used as a selective marker for identifying hybrids. Based on the examination of > 645 700 geminated seeds, the result showed low frequencies (0.05-0.79%) of transgene flow from GM to non-GM rice at close spacing, although with significant variation among mixed-planting proportions. It is concluded that rice transgene flow will occur at a very low frequency (< 1.0%), even if the GM rice is planted at close spacing with non-GM rice, and high densities of GM rice cultivated in the neighborhood of non-GM rice will increase the probability of outcrossing with the non-GM rice. PMID- 16313640 TI - Fructan synthesis is inhibited by phosphate in warm-grown, but not in cold treated, excised barley leaves. AB - The inhibition of fructan accumulation by phosphate was investigated in warm grown and cold-treated barley (Hordeum vulgare) plants. Detached leaves were incubated in water or phosphate for 24 h under lighting or in darkness. Fructosyltransferase, sucrose phosphate synthase (SPS) and cytosolic fructose-1,6 bisphosphatase (FBPase) activities were subsequently analysed, as well as the content of carbohydrates, hexose-phosphates, phosphate, amino acids and protein. In warm-grown leaves, phosphate decreased fructan accumulation and total carbon in carbohydrates and did not affect protein content. Phosphate increased hexose phosphates, phosphate and amino acids. Fructosyltransferase and FBPase activities were not affected by phosphate feeding, while SPS activity was inhibited by phosphate in incubations in both light and darkness. In cold-treated leaves, which before incubation had higher SPS activities than warm-grown leaves, phosphate had no inhibitory effect on fructan accumulation, carbohydrate content or total C in carbohydrates. The activities of SPS and FBPase were unaffected by phosphate. The results indicate that phosphate decreases fructan accumulation through an inhibition of SPS whenever this activity is not high before a rise in phosphate content. PMID- 16313641 TI - Functional role of anthocyanins in high-light winter leaves of the evergreen herb Galax urceolata. AB - High-light leaves of the evergreen herb Galax urceolata exhibit a striking color change from green to red during winter months due to anthocyanin synthesis in outermost mesophyll cells. Here we investigate three possible functions of this color change. To test the hypothesis that anthocyanins function as light attenuators, maximum photosystem II efficiency (F(v)/F(m)) of red and green leaves was measured during and after exposure to wavelengths either strongly or poorly absorbed by anthocyanin. To determine whether anthocyanins elevate radical scavenging capacity, antioxidant activity of red and green leaves was assessed using the alpha,alpha-diphenyl-beta-picrylhydrazyl assay. Nonstructural carbohydrate levels were analyzed to test the hypothesis that anthocyanins function as a carbon sink. Declines in F(v)/F(m) under white and green light were significantly greater for green than red leaves, but were comparable under red light. Anthocyanin content positively correlated with antioxidant activity. Although levels of anthocyanins did not appear to be related to nonstructural carbohydrate concentration, high levels of sugars may be necessary for their photo-induction. Results suggest that anthocyanins function as light attenuators and may also contribute to the antioxidant pool in winter leaves. PMID- 16313642 TI - Daylength and temperature during seed production interactively affect adaptive performance of Picea abies progenies. AB - Adaptive traits in Picea abies (Norway spruce) progenies are influenced by the maternal temperatures during seed production. Here, we have extended these studies by testing the effects of maternal photoperiod and temperature on phenology and frost hardiness on progenies. Using eight phytotron rooms, seeds from three unrelated crosses were made in an environmental 2 x 2 factorial combination of long and short days and high and low temperatures. The progenies were then forced to cease growth rapidly at the end of the first growing season. An interactive memory effect was expressed the second growth season. Progenies from high temperature and short days, and from low temperatures and long days, started growth later in spring, ceased shoot growth later in summer, grew taller and were less frost hardy in the autumn than their full siblings from low temperatures and short days, and from high temperatures and long days. Norway spruce has developed a memory mechanism, regulating adaptive plasticity by photoperiod and temperature, which could counteract harmful effects of a rapidly changing climate. PMID- 16313643 TI - Summer and winter sensitivity of leaves and xylem to minimum freezing temperatures: a comparison of co-occurring Mediterranean oaks that differ in leaf lifespan. AB - Freezing sensitivity of leaves and xylem was examined in four co-occurring Mediterranean oaks (Quercus spp.) grown in a common garden to determine whether freezing responses of leaves and xylem were coordinated and could be predicted by leaf lifespan. Freezing-induced embolism and loss of photosynthetic function were measured after overnight exposure to a range of subzero temperatures in both summer and winter. Both measures were found to be dependent on minimum freezing temperature and were correlated with leaf lifespan and vessel diameter. The dependence of xylem embolism on minimum freezing temperature may result from the decline in water potential with ice temperature that influences the redistribution of water during freezing and leads to an increase in xylem tension. Winter acclimatization had a relatively small effect on the vulnerability to freezing-induced embolism, although leaf photosynthetic function showed a strong acclimatization response, particularly in the two evergreen species. Quercus ilex, the species with the longest leaf lifespan and narrowest vessel diameters, showed the highest freezing tolerance. This helps explain its ability to inhabit a broad range throughout the Mediterranean region. By contrast, the inability of the deciduous oaks to maintain photosynthetic and vascular function throughout the winter indicates a competitive disadvantage that may prevent them from expanding their ranges. PMID- 16313644 TI - Allocation of reserve-derived and currently assimilated carbon and nitrogen in seedlings of Helianthus annuus under sub-ambient and elevated CO growth conditions. AB - Here, we analysed the transition from heterotrophic to autotrophic growth of the epigeal species sunflower (Helianthus annuus), and how transition is affected by CO(2). Growth analysis and steady-state (13)CO(2)/(12)CO(2) and (15)NO(3) ( )/(14)NO(3) (-) labelling were used to quantify reserve- and current assimilation derived carbon (C) and nitrogen (N) allocation to shoots and roots in the presence of 200 and 1,000 micromol CO(2) mol(-1) air. Growth was not influenced by CO(2) until cotyledons unfolded. Then, C accumulation at elevated CO(2) increased to a rate 2-2.5 times higher than in sub-ambient CO(2) due to increased unit leaf rate (+120%) and leaf expansion (+60%). CO(2) had no effect on mobilization and allocation of reserve-derived C and N, even during the transition period. Export of autotrophic C from cotyledons began immediately following the onset of photosynthetic activity, serving roots and shoots near simultaneously. Allocation of autotrophic C to shoots was increased at sub ambient CO(2). The synchrony in transition from heterotrophic to autotrophic supply for different sinks in sunflower contrasts with the sequential transition reported for species with hypogeal germination. PMID- 16313645 TI - Tropospheric O(3) compromises net primary production in young stands of trembling aspen, paper birch and sugar maple in response to elevated atmospheric CO(2). AB - Concentrations of atmospheric CO(2) and tropospheric ozone (O(3)) are rising concurrently in the atmosphere, with potentially antagonistic effects on forest net primary production (NPP) and implications for terrestrial carbon sequestration. Using free-air CO(2) enrichment (FACE) technology, we exposed north-temperate forest communities to concentrations of CO(2) and O(3) predicted for the year 2050 for the first 7 yr of stand development. Site-specific allometric equations were applied to annual nondestructive growth measurements to estimate above- and below-ground biomass and NPP for each year of the experiment. Relative to the control, elevated CO(2) increased total biomass 25, 45 and 60% in the aspen, aspen-birch and aspen-maple communities, respectively. Tropospheric O(3) caused 23, 13 and 14% reductions in total biomass relative to the control in the respective communities. Combined fumigation resulted in total biomass response of -7.8, +8.4 and +24.3% relative to the control in the aspen, aspen birch and aspen-sugar maple communities, respectively. These results indicate that exposure to even moderate levels of O(3) significantly reduce the capacity of NPP to respond to elevated CO(2) in some forests. PMID- 16313646 TI - Soil nutrient heterogeneity interacts with elevated CO2 and nutrient availability to determine species and assemblage responses in a model grassland community. AB - Interactive effects of atmospheric CO(2) concentration ([CO(2)]), soil nutrient availability and soil nutrient spatial distribution on the structure and function of plant assemblages remain largely unexplored. Here we conducted a microcosm experiment to evaluate these interactions using a grassland assemblage formed by Lolium perenne, Plantago lanceolata, Trifolium repens, Anthoxanthum odoratum and Holcus lanatus. Assemblages exhibited precise root foraging patterns, had higher total and below-ground biomass, and captured more nitrogen when nutrients were supplied heterogeneously. Root foraging responses were modified by nutrient availability, and the patterns of N capture by interactions between nutrient distribution, availability and [CO(2)]. Greater above-ground biomass was observed under elevated CO(2) only under homogeneous conditions of nutrient supply and at the highest availability level. CO(2) interacted with nutrient distribution and availability to determine foliar percentage N and below : above-ground biomass ratios, respectively. Interactions between nutrient distribution and CO(2) determined the relative contribution to above-ground biomass of four of the species. The responses of dominant and subordinate species to [CO(2)] were dependent on the availability and distribution of nutrients. Our results suggest that soil nutrient distribution has the potential to influence the response of plant species and assemblages to changes in [CO(2)] and nutrient availability. PMID- 16313647 TI - Monitoring the spatial and temporal dynamics of a community of the tree-root endophyte Phialocephala fortinii s.l. AB - Phialocephala fortinii sensu lato was isolated from Picea abies roots that had been collected from the same 3 x 3-m forest plot in 2001 and 2004, to examine the spatial and temporal dynamics of this fungal community. RFLP analysis was used to define the multilocus haplotype (MLH) of each isolate. Pielou's measure of association and chi(2) tests of independence were employed to examine the randomness of patterns of spatial distribution of MLH observed in 2001 and 2004. Population differentiation between the two samplings was tested using the G(ST) statistic. In 2001, 144 strains of 28 MLH were isolated; in 2004, 139 strains of 29 MLH. Abundant MLH in 2001 also prevailed in 2004, and the same two cryptic species were dominant in both collections. The probability of being isolated in both years increased with increasing frequency of an MLH. The patterns of spatial distribution of most MLH did not differ between years. The G(ST) values indicated identity of the two collections. Communities of P. fortinii sensu lato remain spatially and genetically stable for at least 3 yr. PMID- 16313648 TI - Beta-rhizobia from Mimosa pigra, a newly discovered invasive plant in Taiwan. AB - A total of 191 rhizobial isolates from the root nodules of three geographically separate populations of the invasive plant Mimosa pigra in Taiwan were examined using amplified rDNA restriction analysis, 16S rDNA sequences, protein profiles and ELISA. Of these, 96% were identified as Burkholderia and 4% as Cupriavidus taiwanensis. The symbiosis-essential genes nodA and nifH were present in two strains of Burkholderia (PAS44 and PTK47), and in one of C. taiwanensis (PAS15). All three could nodulate M. pigra. Light and electron microscopy studies with a green fluorescent protein transconjugant variant of strain PAS44 showed the presence of fluorescent bacteroids in M. pigra nodules. These bacteroids expressed the nifH protein, hence this is the first confirmation that Burkholderia is a genuine symbiont of legume nodules. The predominance of Burkholderia in Taiwanese M. pigra suggests that this species may have brought its symbionts from its native South America, rather than entering into association with the Taiwanese Mimosa symbiont C. taiwanensis which so successfully nodulates Mimosa pudica and Mimosa diplotricha. PMID- 16313649 TI - The influence of external nitrogen on carbon allocation to Glomus intraradices in monoxenic arbuscular mycorrhiza. AB - The influence of external nitrogen (N) on carbon (C) allocation and processes related to phosphorus (P) metabolism were studied in monoxenic arbuscular mycorrhiza (AM) cultures of Daucus carota. Fungal hyphae of Glomus intraradices proliferated from colonized roots growing on solid medium into C-free liquid minimal medium with two different N and P levels. Furthermore, we exposed the colonized roots to high or low N availability and then studied the mycelial development. Roots were provided with (13)C-glucose in order to follow the C allocation. The mycelium was analysed for phosphatase activity and transcription levels of two nutrient regulated genes. High N availability to the monoxenic AM root reduced the C allocation to the AM fungus while N availability to the mycelium was important for the upregulation of the fungal inorganic phosphorus (Pi)-transporter GiPT. We found that N availability can regulate nutritional processes in arbuscular mycorrhiza. We conclude that negative impacts of N on AM abundance are caused by reduced C allocation from the plant. Upregulation of the fungal Pi-transporter GiPT indicated that increased N availability might induce P limitation in the mycelium. PMID- 16313650 TI - The uptake, metabolism, transport and transfer of nitrogen in an arbuscular mycorrhizal symbiosis. AB - Nitrogen (N) is known to be transferred from fungus to plant in the arbuscular mycorrhizal (AM) symbiosis, yet its metabolism, storage and transport are poorly understood. In vitro mycorrhizas of Glomus intra-radices and Ri T-DNA-transformed carrot roots were grown in two-compartment Petri dishes. (15)N- and/or (13)C labeled substrates were supplied to either the fungal compartment or to separate dishes containing uncolonized roots. The levels and labeling of free amino acids (AAs) in the extra-radical mycelium (ERM) in mycorrhizal roots and in uncolonized roots were measured by gas chromatography/mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC). Arginine (Arg) was the predominant free AA in the ERM, and almost all Arg molecules became labeled within 3 wk of supplying (15)NH(4) (+) to the fungal compartment. Labeling in Arg represented > 90% of the total (15)N in the free AAs of the ERM. [Guanido-2-(15)N]Arg taken up by the ERM and transported to the intra-radical mycelium (IRM) gave rise to (15)N labeled AAs. [U-(13)C]Arg added to the fungal compartment did not produce any (13)C labeling of other AAs in the mycorrhizal root. Arg is the major form of N synthesized and stored in the ERM and transported to the IRM. However, NH(4) (+) is the most likely form of N transferred to host cells following its generation from Arg breakdown. PMID- 16313651 TI - The high-affinity poplar ammonium importer PttAMT1.2 and its role in ectomycorrhizal symbiosis. AB - One way to elucidate whether ammonium could act as a nitrogen (N) source delivered by the fungus in ectomycorrhizal symbiosis is to investigate plant ammonium importers. Expression analysis of a high-affinity ammonium importer from Populus tremulax tremuloides (PttAMT1.2) and of known members of the AMT1 gene family from Populus trichocarpa was performed. In addition, PttAMT1.2 function was studied in detail by heterologous expression in yeast. PttAMT1.2 expression proved to be root-specific, affected by N nutrition, and strongly increased in a N-independent manner upon ectomycorrhiza formation. The corresponding protein had a K(M) value for ammonium of c. 52 microm. From the seven members of the AMT1 gene family, one gene was exclusively expressed in roots while four genes were detectable in all poplar organs but with varying degrees of expression. Ectomycorrhiza formation resulted in a strong upregulation of three of these genes. Our results indicate an increased ammonium uptake capacity of mycorrhized poplar roots and suggest, together with the expression of putative ammonium exporter genes in the ectomycorrhizal fungus Amanita muscaria, that ammonium could be a major N source delivered from the fungus towards the plant in symbiosis. PMID- 16313653 TI - Evidence for a gene-environment interaction in predicting behavioral inhibition in middle childhood. AB - Gene-environment interactions are presumed to shape human behavior during early development. However, no human research has demonstrated that such interactions lead to stable individual differences in fear responses. We tested this possibility by focusing on a polymorphism in the promoter region of the gene for the serotonin transporter (5-HTT). This polymorphism has been linked to many indices of serotonin activity. Specifically, we tested the hypothesis that an interaction between children's 5-HTT status and maternal reports of social support predicts inhibited behavior with unfamiliar peers in middle childhood. Results were consistent with this hypothesis: Children with the combination of the short 5-HTT allele and low social support had increased risk for behavioral inhibition in middle childhood. PMID- 16313654 TI - Children (but not adults) can inhibit false memories. AB - The role of inhibition in children's (5-, 7-, and 11-year-olds') false memory illusions in the Deese-Roediger-McDermott (DRM) paradigm was examined using a list-wise directed-forgetting procedure. Children studied either a single DRM list (control) or two DRM lists in succession with a directed-remembering instruction or a directed-forgetting instruction between list presentations. The findings indicated that, like adults, children effectively suppressed the output of true memories when given a directed-forgetting instruction. Unlike adults, whose false memories are not attenuated in directed-forgetting conditions, children suppressed false memories at recall in the directed-forgetting condition. Because recognition data indicated that the children did generate false memories regardless of instruction, it appears that although adults' false memories are generated automatically and do not become part of their conscious experience, children's false memories are produced with greater effort and conscious processing, and as a result are easier to suppress at output. PMID- 16313655 TI - Pronouns in marital interaction. AB - Recent studies in social psychology have found that the frequency of certain words in people's speech and writing is related to psychological aspects of their personal health. We investigated whether counts of "self" and "other" pronouns used by 59 couples engaged in a problem-solving discussion were related to indices of marital health. One spouse in each couple had a diagnosis of obsessive compulsive disorder or panic disorder with agoraphobia; 50% of the patients and 40% of their spouses reported marital dissatisfaction. Regardless of patients' diagnostic status, spouses who used more second-person pronouns were more negative during interactions, whereas those who used more first-person plural pronouns produced more positive problem solutions, even when negative behavior was statistically controlled. Moreover, use of first-person singular pronouns was positively associated with marital satisfaction. These findings suggest that pronouns used by spouses during conflict-resolution discussions provide insight into the quality of their interactions and marriages. PMID- 16313656 TI - See the ball, hit the ball. PMID- 16313657 TI - Self-discipline outdoes IQ in predicting academic performance of adolescents. AB - In a longitudinal study of 140 eighth-grade students, self-discipline measured by self-report, parent report, teacher report, and monetary choice questionnaires in the fall predicted final grades, school attendance, standardized achievement-test scores, and selection into a competitive high school program the following spring. In a replication with 164 eighth graders, a behavioral delay-of gratification task, a questionnaire on study habits, and a group-administered IQ test were added. Self-discipline measured in the fall accounted for more than twice as much variance as IQ in final grades, high school selection, school attendance, hours spent doing homework, hours spent watching television (inversely), and the time of day students began their homework. The effect of self-discipline on final grades held even when controlling for first-marking period grades, achievement-test scores, and measured IQ. These findings suggest a major reason for students falling short of their intellectual potential: their failure to exercise self-discipline. PMID- 16313658 TI - Time does not heal all wounds. AB - Cross-sectional studies show that divorced people report lower levels of life satisfaction than do married people. However, such studies cannot determine whether satisfaction actually changes following divorce. In the current study, data from an 18-year panel study of more than 30,000 Germans were used to examine reaction and adaptation to divorce. Results show that satisfaction drops as one approaches divorce and then gradually rebounds over time. However, the return to baseline is not complete. In addition, prospective analyses show that people who will divorce are less happy than those who stay married, even before either group gets married. Thus, the association between divorce and life satisfaction is due to both preexisting differences and lasting changes following the event. PMID- 16313659 TI - Relationships between intergroup contact and prejudice among minority and majority status groups. AB - Considerable research has shown that greater intergroup contact corresponds with lower intergroup prejudice, yet little is known regarding how the relationships between contact and prejudice may vary for members of minority and majority status groups. The present research examined differences in contact-prejudice relationships among members of minority and majority status groups, using data from a larger meta-analytic study of the effects of intergroup contact. Results indicate that the relationships between contact and prejudice tend to be weaker among members of minority status groups than among members of majority status groups. Moreover, establishing Allport's (1954) proposed conditions for optimal intergroup contact significantly predicts stronger contact-prejudice relationships among members of majority status groups, but not among members of minority status groups. Implications of these findings for future research on contact between minority and majority status groups are discussed. PMID- 16313660 TI - Detecting silent pauses in speech: a new tool for measuring on-line lexical and semantic processing. AB - In this study, we introduce pause detection (PD) as a new tool for studying the on-line integration of lexical and semantic information during speech comprehension. When listeners were asked to detect 200-ms pauses inserted into the last words of spoken sentences, their detection latencies were influenced by the lexical-semantic information provided by the sentences. Listeners took longer to detect a pause when it was inserted within a word that had multiple potential endings, rather than a unique ending, in the context of the sentence. An event related potential (ERP) variant of the PD procedure revealed brain correlates of pauses as early as 101 to 125 ms following pause onset and patterns of lexical semantic integration that mirrored those obtained with PD within 160 ms of pause onset. Thus, both the behavioral and the electrophysiological responses to pauses suggest that lexical and semantic processes are highly interactive and that their integration occurs rapidly during speech comprehension. PMID- 16313661 TI - Visual short-term memory load suppresses temporo-parietal junction activity and induces inattentional blindness. AB - The right temporo-parietal junction (TPJ) is critical for stimulus-driven attention and visual awareness. Here we show that as the visual short-term memory (VSTM) load of a task increases, activity in this region is increasingly suppressed. Correspondingly, increasing VSTM load impairs the ability of subjects to consciously detect the presence of a novel, unexpected object in the visual field. These results not only demonstrate that VSTM load suppresses TPJ activity and induces inattentional blindness, but also offer a plausible neural mechanism for this perceptual deficit: suppression of the stimulus-driven attentional network. PMID- 16313662 TI - Using cognitive models to map relations between neuropsychological disorders and human decision-making deficits. AB - Findings from a complex decision-making task (the Iowa gambling task) show that individuals with neuropsychological disorders are characterized by decision making deficits that lead to maladaptive risk-taking behavior. This article describes a cognitive model that distills performance in this task into three different underlying psychological components: the relative impact of rewards and punishments on evaluations of options, the rate that the contingent payoffs are learned, and the consistency between learning and responding. Findings from 10 studies are organized by distilling the observed decision deficits into the three basic components and locating the neuropsychological disorders in this component space. The results reveal a cluster of populations characterized by making risky choices despite high attention to losses, perhaps because of difficulties in creating emotive representations. These findings demonstrate the potential contribution of cognitive models in building bridges between neuroscience and behavior. PMID- 16313663 TI - Unique temporal change is the key to attentional capture. AB - Attentional capture refers to the observation that some events break through and attract one's attention even when one is engaged in a task for which these events are irrelevant. Previous research, focusing primarily on spatial factors, has shown that a new object is more salient in this regard than an abrupt change in an object's features. Here we show that feature changes can be as effective as new objects in capturing attention, provided that they occur during a period of temporal calm. Conversely, both feature changes and new objects are far less effective in capturing attention when they occur simultaneously with other display changes, such as coincident with the initial onset of the display or with small visual transients that occur during a display transition. These results highlight the importance of considering both space and time in studies of attentional capture; the most effective stimulus is unique in both dimensions. PMID- 16313664 TI - Top-down attentional modulation in autistic spectrum disorders is stimulus specific. AB - This study investigated top-down modulation of bottom-up attentional capture in children with autistic spectrum disorders (ASDs) and in a typically developing (TD) comparison group using a spatial-cuing task (Experiment 1) and a series of visual search tasks (Experiment 2) employing task-irrelevant distractors. The effect of color and onset singleton distractors was investigated. The TD group showed similar top-down modulation of color and onset stimuli. The ASD group showed typical top-down modulation of color stimuli, but impaired top-down modulation of onset stimuli. The results suggest that children with ASDs may be impaired at prioritizing dynamic stimuli; this has implications for social processing impairments in ASDs. PMID- 16313665 TI - Secret agents: inferences about hidden causes by 10- and 12-month-old infants. AB - Considerable evidence indicates that preverbal infants expect that only physical contact can cause an inanimate object to move. However, very few studies have investigated infants' expectations about the source of causal power. In three experiments, we found that (a) 10- and 12-month-old infants expect a human hand, and not an inanimate object, to be the primary cause of an inanimate object's motion; (b) infants' expectations can lead them to infer a hidden causal agent without any direct perceptual evidence; and (c) infants do not infer a hidden causal agent if the moving object was previously shown to be capable of self generated motion. PMID- 16313666 TI - Understanding the average probability of replication: comment on Killeen (2005). PMID- 16313667 TI - Probability of replication revisited: comment on "An alternative to Null Hypothesis Significance Tests.". PMID- 16313668 TI - Why replication probabilities depend on prior probability distributions: a rejoinder to Killeen (2005). PMID- 16313669 TI - Replicability, confidence, and priors. PMID- 16313670 TI - Exploring ethical considerations for the use of biological and physiological markers in population-based surveys in less developed countries. AB - BACKGROUND: The health information needs of developing countries increasingly include population-based estimates determined by biological and physiological measures. Collection of data on these biomarkers requires careful reassessment of ethical standards and procedures related to issues of safety, informed consent, reporting, and referral policies. This paper reviews the survey practices of health examination surveys that have been conducted in developed nations and discusses their application to similar types of surveys proposed for developing countries. DISCUSSION: The paper contends that a unitary set of ethical principles should be followed for surveys around the world that precludes the danger of creating double standards (and implicitly lowers standards for work done in developing countries). Global ethical standards must, however, be interpreted in the context of the unique historical and cultural context of the country in which the work is being done. Factors that influence ethical considerations, such as the relationship between investigators in developed and developing countries are also discussed. SUMMARY: The paper provides a set of conclusions reached through this discussion and recommendations for the ethical use of biomarkers in populations-based surveys in developing countries. PMID- 16313671 TI - Comparative levels and time trends in blood pressure, total cholesterol, body mass index and smoking among Caucasian and South-Asian participants of a UK primary-care based cardiovascular risk factor screening programme. AB - BACKGROUND: Individuals of South-Asian origin have a comparatively higher cardiovascular disease burden, but there is uncertainty about whether this is due to differences in risk factor levels and trends. We therefore studied comparative levels and time trends in blood pressure (BP), total cholesterol, body mass index (BMI) and current smoking among UK Caucasian and South-Asian individuals. METHODS: Repeatable cross-sectional survey of men and women aged 35-60 attending for first screening as part of a primary-care based cardiovascular risk factor screening programme 1989 and 1999. RESULTS: Of 34,122 men and 37,294 women participants, 499 men (1.5%)and 381 women (1%) were of South-Asian origin. South Asian men had lower systolic [(-4.91 mmHg (95% Confidence Iterval (CI): -3.58 to 6.23)] and diastolic BP [-2.87 mmHg (-2.02 to -3.72)], with no significant differences in cholesterol and BMI. South-Asian women had lower systolic BP [ 1.77 mmHg, 95% (-0.21 to -3.33)], diastolic BP [-1.87 mmHg (-0.92 to -2.82)], cholesterol [-0.24 mmol/l (-0.08 to -0.39)]; and higher BMI [+0.78 kg/m2 (0.25 to 1.3)]. South-Asian men and women had significantly lower prevalence of self reported current smoking (29.0% and 1.8% respectively). With the exception of self-reported current smoking, between ethnic group risk factor trends were not converging. CONCLUSION: With the exception of women's BMI, South-Asian individuals had either lower or similar levels of the examined cardiovascular risk factor levels, compared with Caucasian individuals. Although time trends in smoking were converging, other risk factors trends were similar between the two ethnic groups. Overall the findings do not support the hypothesis that the relatively high cardiovascular disease burden in UK South-Asians is due to higher levels exposure to the examined risk factors. Other hypotheses, such as higher frequency of diabetes and increased genetic predisposition, require further exploration. PMID- 16313672 TI - The peroxisomal multifunctional protein interacts with cortical microtubules in plant cells. AB - BACKGROUND: The plant peroxisomal multifunctional protein (MFP) possesses up to four enzymatic activities that are involved in catalyzing different reactions of fatty acid beta-oxidation in the peroxisome matrix. In addition to these peroxisomal activities, in vitro assays revealed that rice MFP possesses microtubule- and RNA-binding activities suggesting that this protein also has important functions in the cytosol. RESULTS: We demonstrate that MFP is an authentic microtubule-binding protein, as it localized to the cortical microtubule array in vivo, in addition to its expected targeting to the peroxisome matrix. MFP does not, however, interact with the three mitotic microtubule arrays. Microtubule co-sedimentation assays of truncated versions of MFP revealed that multiple microtubule-binding domains are present on the MFP polypeptide. This indicates that these regions function together to achieve high affinity binding of the full-length protein. Real-time imaging of a transiently expressed green fluorescent protein-MFP chimera in living plant cells illustrated that a dynamic, spatial interaction exits between peroxisomes and cortical microtubules as peroxisomes move along actin filaments or oscillate at fixed locations. CONCLUSION: Plant MFP is associated with the cortical microtubule array, in addition to its expected localization in the peroxisome. This observation, coupled with apparent interactions that frequently occur between microtubules and peroxisomes in the cell cortex, supports the hypothesis that MFP is concentrated on microtubules in order to facilitate the regulated import of MFP into peroxisomes. PMID- 16313673 TI - Impact of the introduction of pneumococcal conjugate vaccine on immunization coverage among infants. AB - BACKGROUND: The introduction of pneumococcal conjugate vaccine (PCV) to the U.S. recommended childhood immunization schedule in the year 2000 added three injections to the number of vaccinations a child is expected to receive during the first year of life. Surveys have suggested that the addition of PCV has led some immunization providers to move other routine childhood vaccinations to later ages, which could increase the possibility of missing these vaccines. The purpose of this study was to evaluate whether introduction of PCV affected immunization coverage for recommended childhood vaccinations among 13-month olds in four large provider groups. METHODS: In this retrospective cohort study, we analyzed computerized data on vaccinations for 33,319 children in four large provider groups before and after the introduction of PCV. The primary outcome was whether the child was up to date for all non-PCV recommended vaccinations at 13 months of age. Logistic regression was used to evaluate the association between PCV introduction and the primary outcome. The secondary outcome was the number of days spent underimmunized by 13 months. The association between PCV introduction and the secondary outcome was evaluated using a two-part modelling approach using logistic and negative binomial regression. RESULTS: Overall, 93% of children were up-to-date at 13 months, and 70% received all non-PCV vaccinations without any delay. Among the entire study population, immunization coverage was maintained or slightly increased from the pre-PCV to post-PCV periods. After multivariate adjustment, children born after PCV entered routine use were less likely to be up to-date at 13 months in one provider group (Group C: OR = 0.5; 95% CI: 0.3-0.8) and were less likely to have received all vaccine doses without any delay in two Groups (Group B: OR = 0.4, 95% CI: 0.3 - 0.6; Group C: OR = 0.5, 95% CI: 0.4 0.7). This represented 3% fewer children in Group C who were up-to-date and 14% (Group C) to 16% (Group B) fewer children who spent no time underimmunized at 13 months after PCV entered routine use compared to the pre-PCV baseline. Some disruptions in immunization delivery were also observed concurrent with temporary recommendations to suspend the birth dose of hepatitis B vaccine, preceding the introduction of PCV. CONCLUSION: These findings suggest that the introduction of PCV did not harm overall immunization coverage rates in populations with good access to primary care. However, we did observe some disruptions in the timely delivery of other vaccines coincident with the introduction of PCV and the suspension of the birth dose of hepatitis B vaccine. This study highlights the need for continued vigilance in coming years as the U.S. introduces new childhood vaccines and policies that may change the timing of existing vaccines. PMID- 16313674 TI - Skin and fat necrosis of the breast following methylene blue dye injection for sentinel node biopsy in a patient with breast cancer. AB - Sentinel lymph node biopsy (SLNB) is a simple technique that uses subdermal or peri-tumoral injection of vital blue dye and/or radioactive isotope to identify the first lymph node(s) draining the primary tumor. It has been shown to accurately predict axillary node status in patients with clinically node negative breast cancer. The SLNB is emerging as a new standard of care in patients with early breast cancer. However, the use of methylene blue (MB) dye can be associated with a number of local complications due to its tissue reactive properties. We report a rare case of skin and fat necrosis followed by a dry gangrene of the skin in a female patient with breast cancer who underwent SLNB localization using peri-tumoral injection of MB dye in another institution. This case and literature review suggest that the use of MB dye for SLNB identification should be avoided and replaced with alternative types of blue dye such as Patent Blue V preferably in conjunction with a radioactive isotope tracer. PMID- 16313675 TI - A preliminary study of neuroSPECT evaluation of patients with post-traumatic smell impairment. AB - BACKGROUND: Most olfactory testings are subjective and since they depend upon the patients' response, they are prone to false positive results. The aim of this study was to use quantitative brain perfusion SPECT in order to detect possible areas of brain activation in response to odorant stimulation in patients with post-traumatic impaired smell in comparison to a group of normal subjects. METHODS: Fourteen patients with post-traumatic impaired smell and ten healthy controls were entered in this prospective study. All subjects underwent brain SPECT after intravenous injection of 740-MBq 99mTc-ECD and 48 hours later, the same procedure was repeated following olfactory stimulus (vanilla powder). RESULTS: In most of seven regions of interest (Orbital Frontal Cortex, Inferior Frontal Pole, Superior Frontal Pole, Posterior Superior Frontal Lobe, Parasagittal Area, Occipital Pole, and Cerebellar area) the post-stimulation quantitative values show increased cortical perfusion being more pronounced in normal volunteers than the anosmic patients (except cerebellar areas and the right occipital pole). Maximal activation was observed in orbitofrontal regions (right+ 25.45% and left +25.47%). CONCLUSION: Brain SPECT is a valuable imaging technique in the assessment of post-traumatic anosmia and could be competitive as an alternative to other imaging techniques, especially when functional MRI is unavailable or unsuitable. However, this procedure may benefit from complementary MRI or CT anatomical imaging. PMID- 16313676 TI - Development and initial testing of a computer-based patient decision aid to promote colorectal cancer screening for primary care practice. AB - BACKGROUND: Although colorectal cancer screening is recommended by major policy making organizations, rates of screening remain low. Our aim was to develop a patient-directed, computer-based decision aid about colorectal cancer screening and investigate whether it could increase patient interest in screening. METHODS: We used content from evidence-based literature reviews and our previous decision aid research to develop a prototype. We performed two rounds of usability testing with representative patients to revise the content and format. The final decision aid consisted of an introductory segment, four test-specific segments, and information to allow comparison of the tests across several key parameters. We then conducted a before-after uncontrolled trial of 80 patients 50-75 years old recruited from an academic internal medicine practice. RESULTS: Mean viewing time was 19 minutes. The decision aid improved patients' intent to ask providers for screening from a mean score of 2.8 (1 = not at all likely to ask, 4 = very likely to ask) before viewing the decision aid to 3.2 afterwards (difference, 0.4; p < 0.0001, paired t-test). Most found the aid useful and reported that it improved their knowledge about screening. Sixty percent said they were ready to be tested, 18% needed more information, and 22% were not ready to be screened. Within 6 months of viewing, 43% of patients had completed screening tests. CONCLUSION: We conclude that a computer-based decision aid can increase patient intent to be screened and increase interest in screening. PRACTICE IMPLICATIONS: This decision aid can be viewed by patients prior to provider appointments to increase motivation to be screened and to help them decide about which modality to use for screening. Further work is required to integrate the decision aid with other practice change strategies to raise screening rates to target levels. PMID- 16313677 TI - Women and postfertilization effects of birth control: consistency of beliefs, intentions and reported use. AB - BACKGROUND: This study assesses the consistency of responses among women regarding their beliefs about the mechanisms of actions of birth control methods, beliefs about when human life begins, the intention to use or not use birth control methods that they believe may act after fertilization or implantation, and their reported use of specific methods. METHODS: A questionnaire was administered in family practice and obstetrics and gynecology clinics in Salt Lake City, Utah, and Tulsa, Oklahoma. Participants included women ages 18-50 presenting for any reason and women under age 18 presenting for family planning or pregnancy care. Analyses were based on key questions addressing beliefs about whether specific birth control methods may act after fertilization, beliefs about when human life begins, intention to use a method that may act after fertilization, and reported use of specific methods. The questionnaire contained no information about the mechanism of action of any method of birth control. Responses were considered inconsistent if actual use contradicted intentions, if one intention contradicted another, or if intentions contradicted beliefs. RESULTS: Of all respondents, 38% gave consistent responses about intention to not use or to stop use of any birth control method that acted after fertilization, while 4% gave inconsistent responses. The corresponding percentages for birth control methods that work after implantation were 64% consistent and 2% inconsistent. Of all respondents, 34% reported they believed that life begins at fertilization and would not use any birth control method that acts after fertilization (a consistent response), while 3% reported they believed that life begins at fertilization but would use a birth control method that acts after fertilization (inconsistent). For specific methods of birth control, less than 1% of women gave inconsistent responses. A majority of women (68% or greater) responded accurately about the mechanism of action of condoms, abstinence, sterilization, and abortion, but a substantial percentage of women (between 19% and 57%) were uncertain about the mechanisms of action of oral contraceptives, intrauterine devices (IUDs), Depo-Provera, or natural family planning. CONCLUSION: Women who believe that life begins at fertilization may not intend to use a birth control method that could have postfertilization effects. More research is needed to understand the relative importance of postfertilization effects for women in other populations, and in relation to other properties of and priorities for birth control methods. However, many women were uncertain about the mechanisms of action of specific methods. To respect the principles of informed consent, some women may need more education about what is known and not known about the mechanisms of action of birth control methods. PMID- 16313678 TI - A review of the psychometric properties of the Health of the Nation Outcome Scales (HoNOS) family of measures. AB - BACKGROUND: The Health of the Nation Outcome Scales was developed to routinely measure outcomes for adults with mental illness. Comparable instruments were also developed for children and adolescents (the Health of the Nation Outcome Scales for Children and Adolescents) and older people (the Health of the Nation Outcome Scales 65+). All three are being widely used as outcome measures in the United Kingdom, Australia and New Zealand. There is, however, no comprehensive review of these instruments. This paper fills this gap by reviewing the psychometric properties of each. METHOD: Articles and reports relating to the instruments were retrieved, and their findings synthesised to assess the instruments' validity (content, construct, concurrent, predictive), reliability (test-retest, inter rater), sensitivity to change, and feasibility/utility. RESULTS: Mostly, the instruments perform adequately or better on most dimensions, although some of their psychometric properties warrant closer examination. CONCLUSION: Collectively, the Health of the Nation Outcome Scales family of measures can assess outcomes for different groups on a range of mental health-related constructs, and can be regarded as appropriate for routinely monitoring outcomes. PMID- 16313679 TI - Sentinel node biopsy should be supplemented by axillary sampling in patients with small breast cancers. AB - Axillary clearance provides important prognostic information but is associated with significant morbidity. Sentinel node biopsy can provide staging .141 patients with node negative early breast cancers-tumour size less than 1.5 cm measured clinically or by imaging had guided axillary sampling (sentinel lymph node biopsy in combination with axillary sampling). Four node axillary sampling improved the detection rate of axillary node metastases by 13.6% as compared to blue dye sentinel node biopsy alone. Positive sampled nodes strongly indicated the likelihood of further metastatic being revealed by axillary dissection (67%). Negative sampled nodes in combination with a positive sentinel node biopsy were associated with a much lower rate of further nodal involvement in the axillary clearance (8%). PMID- 16313680 TI - Reactive oxygen species in spermatozoa: methods for monitoring and significance for the origins of genetic disease and infertility. AB - Human spermatozoa generate low levels of reactive oxygen species in order to stimulate key events, such as tyrosine phosphorylation, associated with sperm capacitation. However, if the generation of these potentially pernicious oxygen metabolites becomes elevated for any reason, spermatozoa possess a limited capacity to protect themselves from oxidative stress. As a consequence, exposure of human spermatozoa to intrinsically- or extrinsically- generated reactive oxygen intermediates can result in a state of oxidative stress characterized by peroxidative damage to the sperm plasma membrane and DNA damage to the mitochondrial and nuclear genomes. Oxidative stress in the male germ line is associated with poor fertilization rates, impaired embryonic development, high levels of abortion and increased morbidity in the offspring, including childhood cancer. In this review, we consider the possible origins of oxidative damage to human spermatozoa and reflect on the important contribution such stress might make to the origins of genetic disease in our species. PMID- 16313681 TI - Interleukin-4 (IL4) and Interleukin-4 receptor (IL4RA) polymorphisms in asthma: a case control study. AB - BACKGROUND: IL4/IL4RA pathway plays an important role in atopy and asthma. Different polymorphisms in IL4 and IL4RA genes have been described. Particularly, -33C>TIL4 and 576Q>RIL4RA SNPs have been independently associated to atopy and asthma. The purpose of this study was to analyse these polymorphisms in a population of patients with a well-characterized asthma phenotype. METHODS: A total of 212 unrelated Caucasian individuals, 133 patients with asthma and 79 healthy subjects without symptoms or history of asthma or atopy and with negative skin prick tests were recruited. Lung function was measured by spirometry and asthma was specialist physician-diagnosed according to the ATS (American Thoracic Society) criteria and classified following the GINA (Global Initiative for Asthma) guidelines. Skin prick tests were performed according to EAACI recommendations. -33C>TIL4 was studied with TaqMan assay and 576Q>RIL4RA by PCR RFLP technique. Hardy-Weinberg equilibrium was analysed in all groups. Dichotomous variables were analysed using chi2, Fisher exact test, Monte Carlo simulation test and odds ratio test. To model the effects of multiple covariates logistic regression was used. RESULTS: No statistically significant differences between the group of patients with asthma and the controls were found when the allele and genotype distribution of -33C>TIL4 and 576Q>RIL4RA polymorphisms were compared. However, the T allele of the -33C>TIL4 SNP was more frequent in patients with persistent asthma. Multivariate analysis adjusted for age and sex confirmed that carriers of allele T had an increased risk of persistent asthma (OR: 2.77, 95%CI: 1.18-6.49; p = 0.019). Analysis of combination of polymorphisms showed that patients carrying both the T allele of -33C>TIL4 and the A allele of 576Q>RIL4RA had an increased risk of asthma. This association was particularly observed in persistent asthma [Fisher's p value = 0.0021, Monte Carlo p value (after 10(4) simulations) = 0.0016, OR:3.39; 95% CI:1.50-7.66]. CONCLUSION: Our results show a trend of association between the genetic combination of the T allele of -33C>TIL4 and the A allele of 576Q>RIL4RA with asthma. This genetic variant was more frequently observed in patients with persistent asthma. As long as this study was performed in a small population, further studies in other populations are needed to confirm these results. PMID- 16313682 TI - Prevalence of food allergy: an overview. AB - At present the only cure for food allergy is to avoid eating the food responsible for the allergy. Thus, food allergy or food hypersensitivity is a disease that is not only of concern to the individual who is affected but also to those involved directly and indirectly in supplying and preparing food for the food-allergic individual, and its impact on society should be evaluated on this basis. It is generally assumed that questionnaire-based studies vastly overestimate the prevalence of food hypersensitivity. The reported perceived prevalence of food hypersensitivity varies from 3.24% to 34.9%, which may be explained partly by the difference in reporting lifetime prevalence compared with point prevalence. However, of more importance is the apparent inverse correlation between response rate and prevalence (the higher the response rate, the lower the perceived prevalence). The three most-recent prevalence studies on food hypersensitivity (one on perceived food hypersensitivity and two on confirmed food hypersensitivity) all report estimates for prevalence of approximately 3%, but their criteria for including subjects as being positive are not identical, although they do overlap. Furthermore, because of differences in methodology there is no definitive information to indicate whether the prevalence of food allergy is increasing. However, the high prevalence of pollen-related food allergy in younger adults in the population suggests that the increase in pollen allergy is also being accompanied by an increase in pollen-related food allergy. PMID- 16313683 TI - Dietary exposure to chemicals within the process of risk assessment: possible applications to substances that may cause allergic reactions. AB - Exposure assessment is one of the key parts of the risk assessment process. This task is crucial when evaluating substances for which only intake of toxicologically-important amounts can lead to adverse health effects. Ideally, dietary exposure to hazardous substances can be assessed by combining data on concentration in all food products with data on their consumption. However, it is considered to be neither cost-effective nor necessary to collect detailed data for every substance, and a stepwise procedure is commonly used to focus resources on the most important issues. Screening methods, designed to look for 'worst case' situations, are first used to target chemicals that might be of health concern for the general population or for certain at-risk groups. The quality of the dietary exposure assessments not only depends on the quality of the data collected, but also on the integration tools used for initial screening or for the eventual more precise estimations. A particular challenge is the evaluation of food allergens and components causing other forms of intolerances, since no reliable data seem to be currently available on the type of exposure (amounts and duration) required to induce a food allergy. A different approach from that used for dietary exposure to other hazardous substances has to be adopted. However, the methodologies (such as those used to collect food consumption data) and databases (in particular, information about food labels) developed in such a context could be useful to investigate the exposure conditions leading to the development of food allergies. PMID- 16313684 TI - Characterisation of immune responses to food allergens in mice. AB - There is considerable interest in the development and evaluation of approaches for the safety assessment of novel foods, and in particular in methods for characterisation of allergenic potential. One strategy that has found favour is a tiered approach in which the potential of novel proteins to induce allergic sensitisation is assessed based on considerations of stability of the protein in a simulated gastric juice and homology with, or structural similarity to, known allergens. Linked to such an approach may be evaluation of serological identity with proteins known to cause allergic disease. With the aim of supplementing such approaches with a more direct measurement of potential allergenic activity, attempts have been made to characterise the quality of immune responses elicited in BALB/c strain mice. Such evaluations comprise measurement of IgG and IgE antibody production and (to a lesser extent) of induced cytokine expression patterns. Investigations to date suggest that in mice proteins provoke variable immune responses, those with the potential to cause allergic sensitisation stimulating IgE (and IgG) antibody production. In contrast, non-allergenic, but nevertheless immunogenic, proteins are associated with IgG antibody responses in the absence of marked IgE production. Consistent with the selective activation of selective type 2 T lymphocyte responses, exposure of mice to allergenic protein is associated with preferential expression of IL-4, -5, -10 and -13. Collectively these data suggest that characterisation of the nature of immune response induced in mice by proteins may provide a useful adjunct or alternative to current strategies for the assessment of allergenic potential. PMID- 16313685 TI - Coeliac disease: a diverse clinical syndrome caused by intolerance of wheat, barley and rye. AB - Coeliac disease is a lifelong intolerance to the gluten found in wheat, barley and rye, and some patients are also sensitive to oats. The disease is genetically determined, with 10% of the first-degree relatives affected and 75% of monozygotic twins being concordant. Of the patients with coeliac disease 95% are human leucocyte antigen (HLA)-DQ2 or HLA-DQ8 positive. Characteristically, the jejunal mucosa becomes damaged by a T-cell-mediated autoimmune response that is thought to be initiated by a 33-mer peptide fragment in A2 gliadin, and patients with this disorder have raised levels of anti-endomysium and tissue transglutaminase antibodies in their blood. Coeliac disease is the major diagnosable food intolerance and, with the advent of a simple blood test for case finding, prevalence rates are thought to be approximately 1:100. Classically, the condition presented with malabsorption and failure to thrive in infancy, but this picture has now been overtaken by the much more common presentation in adults, usually with non-specific symptoms such as tiredness and anaemia, disturbance in bowel habit or following low-impact bone fractures. Small intestinal biopsy is necessary for diagnosis and shows a characteristically flat appearance with crypt hypoplasia and infiltration of the epithelium with lymphocytes. Diet is the key to management and a gluten-free diet effectively cures the condition. However, this commitment is lifelong and many aisles in the supermarket are effectively closed to individuals with coeliac disease. Compliance can be monitored by measuring antibodies in blood, which revert to negative after 6-9 months. Patients with minor symptoms, who are found incidentally to have coeliac disease, often ask whether it is necessary to adhere to the diet. Current advice is that dietary adherence is necessary to avoid the long-term complications, which are, principally, osteoporosis and small bowel lymphoma. However, risk of these complications diminishes very considerably in patients who are on a gluten-free diet. PMID- 16313686 TI - The development of the mucosal immune system pre- and post-weaning: balancing regulatory and effector function. AB - The mucosal immune system fulfils the primary function of defence against potential pathogens that may enter across vulnerable surface epithelia. However, a secondary function of the intestinal immune system is to discriminate between pathogen-associated and 'harmless' antigens, expressing active responses against the former and tolerance to the latter. Control of immune responses appears to be an active process, involving local generation of IgA and of regulatory and/or regulated T lymphocytes. Two important periods of maximum exposure to novel antigens occur in the young animal, immediately after birth and at weaning. In both cases the antigenic composition of the intestinal contents can shift suddenly, as a result of a novel diet and of colonisation by novel strains and species of bacteria. Changes in lifestyles of man, and husbandry of animals, have resulted in weaning becoming much more abrupt than previously in evolution, increasing the number of antigens that must be simultaneously evaluated by neonates. Thus, birth and weaning are likely to represent hazard and critical control points in the development of appropriate responses to pathogens and harmless dietary and commensal antigens. Neonates are born with relatively undeveloped mucosal immune systems. At birth this factor may prevent both expression of active immune responses and development of tolerance. However, colonisation by intestinal flora expands the mucosal immune system in antigen specific and non-specific ways. At weaning antibody to fed proteins can be detected, indicating active immune responses to fed proteins. It is proposed that under normal conditions the ability of the mucosal immune system to mount active responses to foreign antigens develops simultaneously with the ability to control and regulate such responses. Problems arise when one or other arm of the immune system develops inappropriately, resulting in inappropriate effector responses to harmless food proteins (allergy) or inadequate responses to pathogens (disease susceptibility). PMID- 16313687 TI - The canine model of dietary hypersensitivity. AB - IgE-mediated dietary hypersensitivity affects approximately 1% of the canine population. There are no breed associations and < or =50% of the patients are aged <1 year at presentation. The most common causative allergens are beef, chicken, milk, eggs, maize, wheat and soyabean. Affected dogs generally display cutaneous disease and 10-15% of the patients may have concurrent alimentary involvement. Diagnosis is currently based on dietary restriction followed by provocation. Procedures for the detection of serum allergen-specific IgE and IgG antibodies are widely available, but these tests correlate poorly with clinical presentation and dietary testing. Recent studies have demonstrated the allergen specificity of IgE antibodies by immunoblotting and have described blood lymphocyte proliferative responses to food allergens. In addition to investigations of spontaneously-arising dietary hypersensitivity, it has also proved possible to study this disorder experimentally. Small colonies of dogs sensitive to particular dietary proteins have been used to study clinical and serological responses to allergen challenge. Hypersensitivity has been experimentally induced in dogs of an atopic phenotype by repeated subcutaneous injection of alum-adjuvanted dietary allergen during neonatal life. These models have been used to trial a range of modified protein or hydrolysate diets. The dog provides a unique large-animal model for investigation of the immunopathogenesis of human dietary hypersensitivity. The dog is closely related genetically to man and shares environmental disease triggers with man. Spontaneously arising canine dietary hypersensitivity is a good clinical mimic of the human disease, and ability to therapeutically manipulate this adverse response in the dog might lead to benefits for human patients. PMID- 16313688 TI - Probiotics and allergy. AB - Allergy is caused by an immune reaction that is out of all proportion to the antigenic stimuli. Classical allergy is a type I hypersensitivity reaction mediated by the interaction of mast cells (and eosinophils) coated with allergen specific IgE and a cross-linking allergen. The physiological outcome is inflammation commonly displayed by urticaria, rhinitis, vomiting and diarrhoea, depending on the route of allergen entry. In extreme reactions anaphylactic shock can result that may lead to death. Chronic allergic responses most commonly present themselves as asthma and eczema. All these symptoms are the consequence of an imbalanced immune system making an unsuitable response to an environmental or food antigen. On bacterial colonisation of the colon after birth the appropriate microbiological stimuli is essential to redress the balance of the skewed T-helper 2 immune response present in the newborn. This normal interaction between baby and microbes is thought to be compromised in the Western world, with a reduction in bifidobacteria and an increase in clostridial species, particularly in bottle-fed infants. The use of probiotic therapy to prevent allergic disease has been demonstrated in two studies using a probiotic Lactobacillus rhamnosus GG in neonates. A long-term reduction in allergy has been shown in the test group, with lactobacillus reducing the incidence of atopic eczema. Management of allergy through probiotics has also been demonstrated in infants, using lactobacilli to control atopic eczema and cow's milk allergy. Unfortunately, these positive results have not been repeated in studies with older children and young adults. PMID- 16313689 TI - Industrial dimensions of food allergy. AB - Serious attempts to estimate the impact of allergic reactions to foods on public health did not begin until the 1980s. Until about 15 years ago food allergy was considered a minor aspect of food safety. Two developments probably prompted a radical re-appraisal of that situation. The first was the apparently inexorable rise in the prevalence of atopic diseases, of which food allergy forms a part, with its possible consequences highlighted by some well-publicised severe reactions. The second was the growth of genetic modification technology, manifested by the commercialisation of transgenic crops. Each of these developments impacted on the food industry in distinct ways. On the one hand, consumers with food allergies had to be enabled to avoid specific allergens in products formulated with existing ingredients. Food manufacturers therefore had to identify those specific allergens down to trace amounts in all the ingredients forming the product and label or remove them. On the other hand, the introduction of products using ingredients from novel sources required an assessment of the allergenicity of these ingredients as an integral part of safety assurance. The approaches used by the food industry to protect existing consumers who have food allergies and those at potential risk of sensitisation from novel proteins will be illustrated, emphasising how they need to be built into every stage of the life cycle of a product. PMID- 16313690 TI - Review of statutory and voluntary labelling of food allergens. AB - Food allergy represents an increasingly important health problem, with prevalence in Western Europe continuing to rise. While some reactions are mild, others can include life-threatening anaphylactic shock. It is estimated that food allergies affect 1-2% of the adult population and < or =8% of children. Relatively few foods are to blame for a large majority of allergic reactions to food in the UK, with most reactions being to milk, eggs, peanuts (Arachis hypogea), nuts, fish, shellfish, soyabean, sesame (Sesamum indicum L.) and wheat. There is currently no cure for food allergy and the few available treatments are focused on relieving the specific symptoms. Consumers with food allergies and food intolerances rely on food labelling to enable them to make informed choices about the foods they eat. Whilst there have recently been important advances in the labelling of food allergens, these advances relate only to requirements for the labelling of the deliberate use of specified food allergens in foods sold pre-packed. In other areas the development of guidance for food manufacturers and retailers on how to assess the risks of possible allergen cross-contamination during food production and manufacture, and then to determine appropriate advisory labelling, is well advanced. Work to address the issue of how to provide appropriate allergen information for foods sold loose, or in catering establishments, is also in progress. PMID- 16313691 TI - GM organisms and the EU regulatory environment: allergenicity as a risk component. AB - The European Food Safety Authority, following a request from the European Commission, has published a guidance document for the risk assessment of GM plants and derived food and feed to assist in the implementation of provisions of Regulation (EC) 1829/2003 of the European Parliament and Council on GM food and feed. This regulation has applied since 18 April 2004. In principle, hazard identification and characterisation of GM crops is conducted in four steps: characterisation of the parent crop and any hazards associated with it; characterisation of the transformation process and of the inserted recombinant DNA, including an assessment of the possible production of new fusion proteins or allergens; assessment of the introduced proteins (toxicity, allergenicity) and metabolites; identification of any other targetted and unexpected alterations in the GM crop, including changes in the plant metabolism resulting in compositional changes and assessment of their toxicological, allergenic or nutritional impact. In relation to allergenicity specifically, it is clear that this property of a given protein is not intrinsic and fully predictable but is a biological activity requiring an interaction with individuals with a predisposed genetic background. Allergenicity, therefore, depends on the genetic diversity and variability in atopic human subjects. Given this lack of complete predictability it is necessary to obtain, from several steps in the risk-assessment process, a cumulative body of evidence that minimises any uncertainty about the protein(s) in question. PMID- 16313692 TI - Allergenic potential of novel foods. AB - Concerns have been expressed that the introduction of novel foods into the diet might lead to the development of new food allergies in consumers. Novel foods can be conveniently divided into GM and non-GM categories. Decision-tree approaches (e.g. International Life Sciences Institute-International Food Biotechnology Council and WHO/FAO) to assess the allergenic potential of GM foods were developed following the discovery, during product development, of the allergenic potential of GM soyabean expressing a gene encoding a storage protein from Brazil nut (Bertolletia excelsa). Within these decision trees considerations include: the source of the transgene; amino acid homology with known allergens; cross reactivity with IgE from food-allergic individuals; resistance to proteolysis; prediction using animal models of food allergy. Such decision trees are under constant review as new knowledge and improved models emerge, but they provide a useful framework for the assessment of the allergenic potential of GM foods. For novel non-GM foods the assessment of allergenic potential is more subjective; some foods or food ingredients will need no assessment other than a robust protein assay to demonstrate the absence of protein. Where protein is present in the novel non-GM food, hazard and risk assessments need to be made in terms of the quantity of protein that might be consumed, the identity of individual protein components and their relationships to known food allergens. Where necessary, this assessment would extend to serum screening for potential cross reactivities, skin-prick tests in previously-sensitised individuals and double blind placebo-controlled food challenges. PMID- 16313693 TI - Could antioxidant supplementation prevent pre-eclampsia? AB - Pre-eclampsia is a disorder characterised by pregnancy-induced hypertension and new-onset proteinuria occurring in the second half of pregnancy. Worldwide, approximately 2-3% of all pregnant women develop pre-eclampsia. The condition is a major cause of maternal and fetal morbidity and mortality. Abnormal placentation is an important predisposing factor for pre-eclampsia, while endothelial activation appears to be central to the pathophysiological changes, possibly indicative of a two-stage disorder characterised by reduced placental perfusion and a maternal syndrome. There is increasing evidence that pre eclampsia is associated with both increased oxidative stress and reduced antioxidant defences, which has led to the hypothesis that oxidative stress may play an important role in the pathogenesis of pre-eclampsia, perhaps acting as the link in a two-stage model of pre-eclampsia. In support of this hypothesis a small, but important, preliminary study has shown a highly significant (P=0.02) reduction in the incidence of pre-eclampsia in women at risk who were taking a supplement of vitamins C and E from mid-pregnancy. Furthermore, these findings support the hypothesis that oxidative stress is at least partly responsible for the endothelial dysfunction of pre-eclampsia. Several larger multicentre trials are currently underway to evaluate the efficacy, safety and cost benefits of antioxidant supplementation during pregnancy for the prevention of pre-eclampsia in both low- and high-risk women, including women with diabetes. The results of these trials are awaited with interest. PMID- 16313694 TI - Micronutrients in childhood and the influence of subclinical inflammation. AB - In the present paper biomarkers of micronutrient status in childhood and some of the factors influencing them, mainly dietary intake, requirements and inflammation will be examined. On a body-weight basis the micronutrient requirements of children are mostly higher than those of an adult, but most biomarkers of status are not age-related. A major factor that is often overlooked in assessing status is the influence of subclinical inflammation on micronutrient biomarkers. In younger children particularly the immune system is still developing and there is a higher frequency of sickness than in adults. The inflammatory response rapidly influences the concentration in the blood of several important micronutrients such as vitamin A, Fe and Zn, even in the first 24 h, whereas dietary deficiencies can be envisaged as having a more gradual effect on biomarkers of nutritional status. The rapid response to infection may be for protective reasons, i.e. conservation of reserves, or by placing demands on those reserves to mount an effective immune response. However, because there is a high prevalence of disease in many developing countries, an apparently healthy child may well be at the incubation stage or convalescing when blood is taken for nutritional assessment and the concentration of certain micronutrient biomarkers will not give a true indication of status. Most biomarkers influenced by inflammation are known, but often they are used because they are convenient or cheap and the influence of subclinical inflammation is either ignored or overlooked. The objective of the present paper is to discuss: (1) some of the important micronutrient deficiencies in childhood influenced by inflammation; (2) ways of correcting the interference from inflammation. PMID- 16313695 TI - Vitamins and respiratory disease: antioxidant micronutrients in pulmonary health and disease. AB - The lungs are continually exposed to relatively-high O(2) tensions, and as such, in comparison with other organs, they represent a unique tissue for the damaging effects of oxidant attack. At particular times during a lifetime this every day challenge may increase exponentially. The first oxidative insult occurs at birth, when cells are exposed to a sudden 5-fold increase in O(2) concentration. Thereafter, the human lung, from infancy through to old age, can be subjected to deleterious oxidative events as a consequence of inhaling environmental pollutants or irritants, succumbing to several pulmonary diseases (including infant and adult respiratory distress syndromes, asthma, chronic obstructive pulmonary disease, cystic fibrosis and cancer) and receiving treatment for these diseases. The present paper will review the concept that consumption of a healthy diet and the consequent ability to establish and then maintain adequate micronutrient antioxidant concentrations in the lung throughout life, and following various oxidative insults, could prevent or reduce the incidence of oxidant-mediated respiratory diseases. Furthermore, the rationale, practicalities and complexities of boosting the antioxidant pool of the respiratory-tract lining fluid in diseases in which oxidative stress is actively involved, by direct application to the lung v. dietary modification, in order to achieve a therapeutic effect will be discussed. PMID- 16313696 TI - Selenium in cancer prevention: a review of the evidence and mechanism of action. AB - Se is an unusual trace element in having its own codon in mRNA that specifies its insertion into selenoproteins as selenocysteine (SeCys), by means of a mechanism requiring a large SeCys-insertion complex. This exacting insertion machinery for selenoprotein production has implications for the Se requirements for cancer prevention. If Se may protect against cancer, an adequate intake of Se is desirable. However, the level of intake in Europe and some parts of the world is not adequate for full expression of protective selenoproteins. The evidence for Se as a cancer preventive agent includes that from geographic, animal, prospective and intervention studies. Newly-published prospective studies on oesophageal, gastric-cardia and lung cancer have reinforced previous evidence, which is particularly strong for prostate cancer. Interventions with Se have shown benefit in reducing the risk of cancer incidence and mortality in all cancers combined, and specifically in liver, prostate, colo-rectal and lung cancers. The effect seems to be strongest in those individuals with the lowest Se status. As the level of Se that appears to be required for optimal effect is higher than that previously understood to be required to maximise the activity of selenoenzymes, the question has been raised as to whether selenoproteins are involved in the anti-cancer process. However, recent evidence showing an association between Se, reduction of DNA damage and oxidative stress together with data showing an effect of selenoprotein genotype on cancer risk implies that selenoproteins are indeed implicated. The likelihood of simultaneous and consecutive effects at different cancer stages still allows an important role for anti-cancer Se metabolites such as methyl selenol formed from gamma-glutamyl selenomethyl-SeCys and selenomethyl-SeCys, components identified in certain plants and Se-enriched yeast that have anti-cancer effects. There is some evidence that Se may affect not only cancer risk but also progression and metastasis. Current primary and secondary prevention trials of Se are underway in the USA, including the Selenium and Vitamin E Cancer Prevention Trial (SELECT) relating to prostate cancer, although a large European trial is still desirable given the likelihood of a stronger effect in populations of lower Se status. PMID- 16313697 TI - Micronutrients: dietary intake v. supplement use. AB - Whilst clinical deficiency of micronutrients is uncommon in the developed world, a suboptimal intake of certain micronutrients has been linked with an increased risk of chronic diseases such as CVD and cancer. Attention has therefore focused on increasing micronutrient status in order to theoretically reduce chronic disease risk. Increasing micronutrient status can involve a number of approaches: increasing dietary intake of micronutrient-rich foods; food fortification; use of supplements. Observational cohort studies have demonstrated an association between high intakes of micronutrients such as vitamin E, vitamin C, folic acid and beta-carotene, and lower risk of CHD, stroke and cancer at various sites. However, randomised intervention trials of micronutrient supplements have, to date, largely failed to show an improvement in clinical end points. The discordance between data from cohort studies and the results so far available from clinical trials remains to be explained. One reason may be that the complex mixture of micronutrients found, for example, in a diet high in fruit and vegetables may be more effective than large doses of a small number of micronutrients, and therefore that intervention studies that use single micronutrient supplements are unlikely to produce a lowering of disease risk. Studies concentrating on whole foods (e.g. fruit and vegetables) or diet pattern (e.g. Mediterranean diet pattern) may be more effective in demonstrating an effect on clinical end points. The present review will consider the clinical trial evidence for a beneficial effect of micronutrient supplements on health, and review the alternative approaches to the study of dietary intake of micronutrients. PMID- 16313698 TI - Fatty acids and CHD. AB - During the last century much evidence has accumulated to suggest that from a public health perspective the type of fat is more important than the amount of fat. Saturated and trans-fatty acids increase and both n-6 and n-3 PUFA decrease the risk of CHD. Most of the knowledge about the effects of dietary fatty acids on CHD risk is based on observational studies and controlled dietary experiments with intermediate end points (e.g. blood lipoprotein fractions). Information from high-quality randomised controlled trials on fatty acids and CHD is lacking. The Netherlands Institute for Public Health has calculated the potential health gain that can be achieved if the fatty acid composition of the current Dutch diet is replaced by the recommended fatty acid composition. The recommendations of The Netherlands Health Council are: saturated fatty acids <10% energy intake; trans fatty acids <1% energy intake; fish consumption (an indicator of n-3 PUFA) once or twice weekly. Implementation of this recommendation could reduce the incidence of CHD in The Netherlands by about 25,000/year and the number of CHD-related deaths by about 6000/year and increase life expectancy from age 40 years onwards by 0.5 year. These projections indicate the public health potential of interventions that modify the fatty acid composition of the diet. PMID- 16313699 TI - Micronutrients and Alzheimer's disease. AB - The current high life expectancy is overshadowed by neurodegenerative illnesses that lead to dementia and dependence. Alzheimer's disease (AD) is the most common of these conditions, and is considered to be a proteinopathy, with amyloid-beta42 as a key factor, leading via a cascade of events to neurodegeneration. Major factors involved are oxidative stress, perturbed Ca homeostasis and impaired energy metabolism. Protection against oxidative stress by micronutrients (including secondary bioactive substances) has been shown in transgenic Alzheimer model systems to delay AD. Epidemiological evidence is less conclusive, but the vast majority of the evidence supports a protective effect on cognitive functions in old age and AD. Thus, a diet rich in fruits and vegetables but also containing meat and fish is the most suitable to provide adequate micronutrients. The strong link between cardiovascular risk and AD may be explained by common pathogenetic mechanisms mediated, for example, by homocysteine and thus dependant on B vitamins (folate and vitamins B(12) and B(6)). However, micronutrients may also be harmful. The high affinity of amyloid for metals (Fe, Al and Zn) favours the generation of reactive oxygen species and triggers an inflammatory response. Micronutrients in a balanced diet have a long-lasting, albeit low, protective impact on brain aging, hence prevention should be life long. PMID- 16313700 TI - The interrelationship between diet and oral health. AB - Diet and nutrition impact on many oral diseases, in particular dental caries. Consumption of fluoridated water coupled with a reduction in non-milk extrinsic sugar intake is an effective means of caries prevention. However, studies on the fluoride concentration of bottled waters suggest increased consumption of these waters, in preference to fluoridated tap water, would lead to a marked decrease in caries protection. Concerns have been raised about the bioavailability of fluoride from artificially-fluoridated water compared with naturally-fluoridated water. This issue has been addressed in a human experimental study that has indicated that any differences in fluoride bioavailability are small compared with the naturally-occurring variability in fluoride absorption. Research has unequivocally shown sugars to be the main aetiological factor for dental caries, and information on intakes guides health promotion. Repeat dietary surveys of English children over three decades indicate that levels of sugars intake have remained stable, while sources of sugars have changed considerably, with the contribution from soft drinks more than doubling since 1980. Dental caries eventually leads to tooth loss, which in turn impairs chewing ability causing avoidance of hard and fibrous foods including fruits, vegetables and whole grains. A very low intake (<12 g/d) of NSP and fruit and vegetables has been found in edentulous subjects. Provision of prostheses alone fails to improve diet. However, initial studies indicate that customised dietary advice at the time of denture provision results in increased consumption of fruits and vegetables, and positive movement through the stages of change. Feasible means of integrating dietary counselling into the dental setting warrants further investigation. PMID- 16313702 TI - Effect of lithium and lithium withdrawal on potassium-evoked dopamine release and tyrosine hydroxylase expression in the rat. AB - The mood stabilizer lithium is used successfully in the treatment of bipolar affective disorder. However, treatment compliance is frequently poor and sudden withdrawal from lithium therapy is associated with a significantly increased risk of rebound mania. In this study we have used rodents to identify neurobiological changes in dopamine function occurring during chronic lithium treatment and withdrawal from chronic lithium treatment. Rats were maintained for 28 d on a lithium diet or a control diet. A subgroup had their lithium diet substituted with a control diet from day 25 of the treatment period. In-vivo microdialysis was used to study both basal dopamine release and potassium-evoked dopamine released in the shell of the nucleus accumbens. In-situ hybridization histochemistry was used to study the abundance of mRNA coding for dopamine's synthetic enzyme, tyrosine hydroxylase in the ventral tegmental area. Basal dopamine levels did not differ across any of the three treatment groups. However, the potassium-evoked dopamine release was significantly attenuated in lithium and lithium-withdrawn rats compared to control rats. Tyrosine hydroxylase mRNA abundance in the ventral tegmental area did not differ between any of the three treatment groups. These data suggest that decreased dopamine release may mediate the mood stabilizing action of lithium. However, in this paradigm a rebound increase in dopamine release was not found after withdrawal from lithium treatment. PMID- 16313703 TI - Psychosis associated with elevated trough tacrolimus blood concentrations after combined kidney-pancreas transplant. PMID- 16313704 TI - Neuroleptic malignant syndrome induced by perospirone. PMID- 16313705 TI - Atypical antipsychotic drugs and organization of long-term semantic memory: multidimensional scaling and cluster analyses of category fluency performance in schizophrenia. AB - Organization of semantic memory, one of the domains of cognitive function, is impaired in patients with schizophrenia, and is predictive of functional outcomes. The Category Fluency Task (CFT) has been used to evaluate organization of long-term semantic memory by means of visualizing semantic associations in the form of 'cognitive map' and cluster structures. While atypical antipsychotic drugs (AAPDs) have been shown to ameliorate overall cognitive deficits, little is known about the efficacy of AAPDs for improving higher cognitive functions, such as semantic memory organization. The purpose of the present study was to determine if treatment with olanzapine or ziprasidone has beneficial influence on organization of semantic memory, as revealed by analysis of data from the CFT, in patients with schizophrenia. A retrospective analysis of an open-label trial was conducted for 33 patients with schizophrenia who were treated with either olanzapine or ziprasidone. Nineteen subjects were unmedicated at baseline. The CFT and Letter Fluency Task, as well as the Brief Psychiatric Rating Scale (BPRS) and Quality of Life Scale (QLS), were administered at baseline and 6 wk of the treatment. Semantic structures were obtained by multidimensional scaling analysis and hierarchical cluster analysis of verbal outputs from the CFT. At baseline, no meaningful dimension or cluster was observed in the semantic structure; however, knowledge-based dimensions (wild vs. domestic) appeared after treatment with olanzapine or ziprasidone. Cluster structures also became organized, especially after treatment with olanzapine. Scores of QLS, but not those of BPRS, improved during treatment with the AAPDs. These results suggest a facilitative influence of AAPDs on higher cognitive functions, such as organization of semantic memory, in patients with schizophrenia. PMID- 16313706 TI - 360-degree retinectomy for severe ocular rupture. AB - OBJECTIVE: To investigate the effect of 360-degree retinectomy for severe ocular rupture and evaluate the related factors associated with prognosis. METHODS: A retrospective analysis was performed on 28 consecutive patients, 18 men and 10 women, with severe ocular rupture admitted to the Chinese PLA General Hospital between January 1999 and January 2002. The patients' ages ranged from 16 to 58 years, mean 29.5 years. All operations were performed by the first author. All patients underwent standard 3-port pars plana vitrectomy with scleral encircling buckle, endophotocoagulation, and tamponaded with silicone oil. Lensectomy were performed on 15 eyes. 360-degree retinotomy and retinectomy were performed at anterior equator on 16 eyes, and posterior equator on 12 eyes. RESULTS: During operation, all patients were confirmed to have vitreous hemorrhage and retinal detachment, in which hemorrhagic ciliary body detachment was found in 9 eyes, suprachoroidal hemorrhage in 12 eyes, retinal twisting like flower stalk in 11 eyes, and retinal incarceration in 17 eyes. Postoperative follow-up was conducted in 26 cases, ranged from 6 months to 46 months. Retinal reattachment achieved in 20 eyes (76.9%), in which three patients' retina was reattached after revitrectomy. The silicone oil was removed at the fourth month in 8 eyes postoperatively. Corrected visual acuity of 0.02 or more was in 14 eyes (70.0%) among the 20 patients with reattached retina. Visual acuity of 0.05 or more was in 7 eyes (35.0a), and 0.1 or more was in 3 eyes. The best visual acuity was 0.3 in 1 eye. CONCLUSIONS: Retinectomy may improve the prognosis of severe ocular rupture and save the visual function of some patients. PMID- 16313707 TI - Surgical treatment of infected arterial injuries of the major limb. AB - OBJECTIVE: To analyze retrospectively the surgical treatment of 21 cases of infected arterial injuries of the major limb hospitalized from 1989 to 2003 in our department. METHODS: After a radical debridement and drainage, an autologous vessel bypass reconstruction, simple vessel ligation or amputation were carried out respectively according to each patient's condition. In order to make a better milieu for the grafts, local musculo-cutaneous flaps were translocated to cover the wounds in 9 cases. RESULTS: 1 case (4.8%) died during the treatment. 5 cases (5/21, 23.8%) underwent amputations. Blood supply reconstruction was successful in 15 cases (15/21, 71.4%). 93.3% (14/15) of the cases were successful in getting an unobstructed vessel at early stage. Thrombosis occurred in 1 case (1/15, 6.7%) and a sound result was obtained after a second operation. CONCLUSIONS: Radical debridement combined with bypass vessel reconstruction with autologous vein and with musculo-cutaneous flap translocation is an effective treatment for infective limb vascular injuries. PMID- 16313708 TI - Ontogeny of expression of basic fibroblast growth factor and its receptors in human fetal skin. AB - OBJECTIVE: To investigate the expression characteristics of basic fibroblast growth factor (bFGF) and its receptors, flg (FGFR1) and bek (FGFR2), in fetal skin at different gestational ages underlying the relevance of these 3 proteins to skin development and the mechanisms underlying the phenotypic transition from scarless to scar-forming healing. METHODS: Eighteen specimens of fetal skin biopsies of human embryo were obtained from spontaneous abortions at different gestational ages of 13-32 weeks. Gene expression of bFGF, bek and flg was examined with reverse transcription-polymerase chain reaction (RT-PCR). The dynamic expression and distribution of these 3 proteins were detected with streptavidin peroxidase (SP) immunohistochemical staining method. RESULTS: In the early gestational fetal skin, genes of bFGF and flg were strongly expressed and more protein contents of these 2 proteins were found as compared with the genes at late gestation fetal skin (2.446+/-0.116 and 2.066+/-0.152 versus 2.157+/ 0.101 and 1.818+/-0.086, respectively, P<0.05). On the contrary, the levels of gene expression and protein content of bek were not differently expressed in the early gestational fetal skin versus the late ones. Protein particles of bFGF were mainly distributed in the epidermal cells and some fibroblasts. Bek was mainly located in the cell membrane and cytoplasm of epidermal cells while flg protein was principally located in the epidermal cells, endothelial cells and some fibroblasts. CONCLUSIONS: The endogenous bFGF and their receptors might be involved in the cutaneous development at fetal stage. The differently expressing levels of bFGF and flg during gestation may be related to scarless or scar forming repair during gestation. PMID- 16313709 TI - Effect of human neural progenitor cells on injured spinal cord. AB - OBJECTIVE: To study whether human neural progenitor cells can differentiate into neural cells in vivo and improve the recovery of injured spinal cord in rats. METHODS: Human neural progenitor cells were transplanted into the injured spinal cord and the functional recovery of the rats with spinal cord contusion injury was evaluated with Basso-Beattie-Bresnahan (BBB) locomotor scale and motor evoked potentials. Additionally, the differentiation of human neural progenitor cells was shown by immunocytochemistry. RESULTS: Human neural progenitor cells developed into functional cells in the injured spinal cord and improved the recovery of injured spinal cord in both locomotor scores and electrophysiological parameters in rats. CONCLUSIONS: Human neural progenitor cells can treat injured spinal cord, which may provide a new cell source for research of clinical application. PMID- 16313710 TI - Diagnosis and treatment of diffuse axonal injury in 169 patients. AB - OBJECTIVE: To evaluate current diagnosis and therapeutic effect and outcome of diffuse axonal injury (DAI) in 169 patients. METHODS: The data of 169 DAI patients treated in the Second, Sixth, Eighth and Ninth Hospitals of Shenzhen and Shekou Hospital from January 2001 to January 2005 were collected. The imaging features, classification, GCS (Glasgow coma scale), treatment and outcome of the 169 patients were retrospectively analyzed. RESULTS: The simpler the imaging features, the closer the focus of DAI to the periphery of hemisphere and the higher the GCS score, the better the prognoses of DAI patients will be. CONCLUSIONS: The prognoses of DAI patients are closely related to the imaging features and classification, GCS and clinical treatment. PMID- 16313711 TI - The expression of N-cadherin, fibronectin during chondrogenic differentiation of MSC induced by TGF-beta(1). AB - OBJECTIVE: To investigate the role of N-cadherin and fibronectin during chondrogenesis. METHODS: Immunohistochemical method and autibody induced changes of aggregation of cells were used to assay the expressions of N-cadherin and fibronectin during cell differentiation. RESULTS: The N-cadherin was present in the area of the cell nodular area in the 24 hours group after adding chondrogenic revulsant, then there was a down-regulating trend. Fibronectin was expressed in 48 and 72 hours groups after adding chondrogenic revulsant, and showed to be negative afterward. The antibody against fibronectin or N-cadherin could inhibit the formation of cellular nodule markedly. CONCLUSIONS: Cell adhesion factors play an important role during cell differentiation. TGF-beta(1) stimulates chondrogenesis via transition from an initial N-cadherin-contributing stage to a succedent fibronectin-contributing stage during the process of chondrogenesis in MSCs. Further study is needed to evaluate whether or not it can promote chondrogenesis by transfecting cDNA of CAMs to MSCs. PMID- 16313712 TI - Silencing effect of recombinant plasmids PPARgamma-pSUPER-EGFP on the expression of peroxisome proliferator-activated receptor gamma. AB - OBJECTIVE: To investigate the silencing effect of gene encoding peroxisome proliferator-activated receptor gamma (PPARgamma) on the expression of tumor necrosis factor alpha (TNFalpha) by constructing vectors for RNA interference in RAW264.7 cells. METHODS: The pSUPER-EGFP vectors were used to transcribe functional small interfering RNA (siRNA). Four pairs of oligonucleotides (64 nt) targeting PPARgamma gene were inserted into the downstream of the H1 promotor, with their veracity confirmed by double digestion and sequencing. Western blotting and immunofluorescence assay were used to examine the silencing effect of PPARgamma gene in RAW264.7 cells. Meanwhile, the TNFalphalevel was determined by Sandwich ELISA. RESULTS: Compared with other recombinant pSUPER-EGFP vectors (R-pSUPER.EGFP), R-pSUPER.EGFP2 induced the best silencing effect on the expression of PPARgamma in RAW264.7 cells, which played an obvious inhibitory role in down-regulating the TNFalphaexpression after the curcumin and lipopolysaccharide (LPS) stimulation. CONCLUSIONS: PPARgamma-pSUPER-EGFP inducing a silencing effect on the expression of PPARgamma can efficiently play a negative role in controlling the inflammatory responses of RAW264.7 cells. PMID- 16313713 TI - Femoral component revision using extensively porous-coated cementless stem. AB - OBJECTIVE: To evaluate the clinical and radiographic results of extensively porous-coated femoral stem in revision of total hip arthroplasty (THA). METHODS: From January 1999 to December 2003, fifteen hips of fifteen cases received revision of THA with extensively porous-coated femoral stem. There were six males and nine females. The average age was 66 years (ranging 58-82 years). The reason for the revision was aseptic loosening in 10 cases, septic loosening in 2, femoral shaft fracture around loose implant in 2, and femoral revision for malposition of the femoral component in 1. All the patients were clinically evaluated using Harris hip score and radiographically evaluated both preoperatively and postoperatively at regular follow-up intervals. RESULTS: No patients were lost for follow-up. The average length of follow-up was 2.3 years (range, 1-5 years). The average preoperative Harris hip score was 42 points, which was improved to 89 points at latest follow-up. The latest follow-up showed that bone in-growth occurred in fourteen stems and solid fibrous fixation in one. Complications consisted of femoral shaft fracture in two cases (1 undisplaced distal femur fracture and 1 cortical perforation at the tip of the prosthesis), and postoperative dislocation in one. There was no mechanical failure of the stem in this study. CONCLUSIONS: Satisfactory results of short-term clinical and radiographic follow-up have been achieved in using extensively porous-coated femoral stem for revision of THA. It should be noticed that the straight, 203 mm stem should be used with caution in short people. PMID- 16313714 TI - Epidemiology of assaultive injuries in areas of Sichuan province, China. AB - OBJECTIVE: To scrutinize the epidemiological characteristics of assaultive injuries in Sichuan province, China. METHODS: A survey of all cases of assaultive injuries reported by police was performed during 8 years in eight counties of Sichuan province, China. A total of 2862 victims and 2856 offenders were registered. RESULTS: The majority of victims and offenders were young men at the age of 20-39 and only received an education at secondary school or primary school. The largest fraction of these cases took place at farm or by-place during 10.00-11.00 o'clock, 16.00-17.00 o'clock and 20.00-21.00 o'clock. The tangles caused by trifles were the most common factors inducing assaultive injuries and accounted for 42.1 percent of the causes of assaults. Blunt injuries were mainly caused by punching (40%) and kicking (17.2%). About 37.3% of the lesions seriously happened in the regions of face and head. Open wounds accounted for 40.3% of these different injuries. CONCLUSIONS: It is valuable to take some specific measures to prevent and control assaultive injuries according to their territorial characteristics. PMID- 16313715 TI - Effects of chitosan on cell proliferation and collagen production of tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes. AB - OBJECTIVE: To study the proliferation and collagen production of tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes; and the effects of chitosan on cell proliferation and collagen production in the 3 cell types of rabbit flexor tendon. METHODS: Three cell lines of tendon sheath, epitenon, and endotenon were isolated from rabbit flexor tendon and cultured. Cell culture media was added with chitosan. The cell number and production of collagens I, II, and III were measured and compared with those cultured without chitosan. The expression of type I collagen in tendon sheath fibroblasts was determined by quantitative analysis of reverse-transcription polymerase chain reaction. RESULTS: All 3 cell lines produced collagens I, II, and III. Adding chitosan to cell media resulted in a significant decrease in cell number in all 3 cell lines. In addition, there was a significant decrease in collagens I, II, and III production in all 3 cell lines as well as the expression levels of type I collagen in tendon sheath fibroblasts (P<0.05). CONCLUSIONS: Chitosan can inhibit cell proliferation and collagen production of the tendon sheath, epitenon, and endotenon, and may provide a promising approach to obviating tendon adhesion formation clinically. PMID- 16313716 TI - Simultaneous reconstruction of anterior cruciate ligament and posterior cruciate ligament by using allogeneic patellar tendon under arthroscopy. AB - OBJECTIVE: To evaluate operative effects of a new method to reconstruct anterior crucial ligament (ACL) and posterior crucial ligament (PCL) simultaneously by using patellar tendon under arthroscopy. METHODS: From November 1999 to November 2003, the injured ACL and PCL of 11 patients were fixated with compressed screws and reconstructed under arthroscopy with the bone-patellar tendon-bone treated with deep hypothermia andgamma radiation. At the same time, 2 patients were treated with medial collateral ligament (MCL) reconstruction, 3 with lateral collateral ligament (LCL) reconstruction, 1 with meniscus suture and 4 with whole or partial resection. RESULTS: All patients were followed up for 12-26 months (average 16.5 months). The Lysholm score method was employed to evaluate the knee function. The average preoperative score was 45.3 and the postoperative score was 86.4. Anterior drawer test (ADT) was positive in 11 knees preoperatively and feeble positive in one knee postoperatively. Lachman test was positive in 11 knees preoperatively and in one postoperatively, and feebly positive in two postoperatively. Posterior drawer test was positive in 11 knees preoperatively and feebly positive in 2 postoperatively. There were 2 knees with tolerable pain and 2 with knee flexion of 5-20. CONCLUSIONS: As for simultaneous reconstruction of ACL and PCL under arthroscopy, allogeneic bone-patellar tendon-bone can not only avoid injury and complication caused by autografting, but also help rehabilitation of the knee function. PMID- 16313717 TI - The applications of arthroscopy on malleolus fractures. AB - It was well known that in 1918 Takagi performed the first arthroscopic inspection of a cadaver's knee in Japan.(1) His interest in this area laid the foundation for arthroscopy and facilitated the development of arthroscope. In 1931, Burman reported an experimental study on the arthroscopic exploration of cadaveric joints, but he believed that the ankle joint was unsuitable for such techniques because it was too narrow to pass through the posterior puncture.(2) Unexpectedly, several years later Takagi described a routine method for arthroscopic examination of the ankle.(1) Since 1970's, ankle arthroscopy had made some significant progress after almost four decades of silence. In 1972, Watanabe reported 28 cases adopting his newly-developed fiberoptic arthroscope and described the anteromedial, anterolateral and posterior approaches.(3) Then, in 1976, Chen reviewed his experience with ankle arthroscopy on 67 patients and 17 cadavers. He elaborately analyzed the various compartments within the ankle and described their anatomy in detail.(4) Subsequently, many authors reported their experiences and techniques in this field.(5-9) In 2000, Hintermann addressed his experience of the arthroscopic application in acute fractures of the ankle.(10) Meanwhile, with rich knowledge about the anatomic portals, some advanced technologies, including video camera, fiberoptic light transmission, joint distraction by invasive or non-invasive means and instruments for small joints, make it possible to perform diagnostic and operative arthroscopy in the ankle. PMID- 16313718 TI - Foreign body in the heart. AB - As an emergency, a foreign body in the heart should generally be removed as early as possible by surgical or non-surgical procedures in order to avoid any complications. Early treatment typically results in a better prognosis. PMID- 16313719 TI - [A study on the inhibitory effect on transplanted tumor growth expressing HCV NS3 protein by HCV minigene vaccine based on invariant chain substitution in mice]. AB - OBJECTIVES: To establish a transplanted tumor producing HCV NS3 protein in mice and study the therapeutic effect of minigene vaccine based on invariant chain substitution. METHODS: SP2/0-NS3 cells expressing HCV NS3 antigen were injected subcutaneously into BALB/ c mice. After three days of inoculation, different therapeutical reagents were injected intramuscularly into different groups of mice. The boost immunization was carried out two weeks after the first immunization. The efficiency of HCV NS3 Th1 minigene vaccine was estimated after 60 days observation. RESULTS: For saline, pCI-neo, pHCV-NS3 and pHCV-NS3-Th1 treated groups, the induction period needed for tumor growth was 16.17+/-2.55, 14.40+/-1.82, 16.75+/-2.36, and 24.00+/-5.57 days (t =2.623, P =0.034 vs saline, t =3.713, P =0.010 vs pCI-neo and t =2.425, P =0.045 vs pHCV-NS3) respectively. The tumorigenesis rates were 100%, 100%, 57.1% (8/14, chi2 = 6.190, P = 0.013 vs saline and chi2 = 6.608, P = 0.010 vs pCI-neo) and 46.7% (7/15, chi2 = 9.707, P = 0.002 vs saline and chi2 = 10.311, P = 0.001 vs pCI-neo ) respectively. The survival rates were 0, 0, 50.0% (7/14, chi2 = 5.787, P = 0.016 vs saline and chi2 = 9.333, P = 0.002 vs pCI-neo) and 53.3% (8/15, chi2 = 6.651, P = 0.010 vs saline and chi2 = 10.311, P = 0.001 vs pCI-neo) respectively. The average tumor diameter of the pHCV-NS3-Th1 treated group was significantly smaller compared with the control groups and the pHCV-NS3 treated group (P =0.001). Moreover, the average survival time of tumor-bearing mice immunized with pHCV-NS3-Th1 was 6 days longer compared with the saline treated group, 12 days longer compared with the pCI-neo treated group (P =0.001), and 6 days compared with the pHCV-NS3 treated group. CONCLUSION: HCV NS3 Th1 epitope vaccine might be a potential biotherapy candidate against HCV infection. PMID- 16313720 TI - [Infection of tupaia hepatocytes with hepatitis C virus in vitro]. AB - OBJECTIVE: Tupaia belangeri (tree shrew) has a close phylogenetic relationship with primates and has been shown to be susceptible to a variety of human viruses. This study was conducted to investigate whether or not hepatitis C virus (HCV) could infect primary tupaia hepatocytes (PTHs) in vitro. METHODS: Serum-derived HCV was cultivated with PTHs, and then positive and negative strand HCV RNA in PTHs, as well as the encapsidated HCV RNA in the culture medium were detected to evaluate the infection. Virus from the culture medium of the infected PTHs was passed to naive PTHs, and the quasispecies of HCV were compared among the inoculum and PTHs after infection and passage. RESULTS: Both positive and negative strand HCV RNA were detected in PTHs after infection. The negative strand RNA was detectable from day 5 to day 10 after infection, while the positive strand RNA was positive up to day 14. HCV RNA, which was RNase resistant, could be detected from the culture medium of the infected PTHs from day 3 to day 14. Production of infectious virons of PTH were demonstrated by passage HCV to naive PTHs. Compared analysis of HCV quasispecies after infection and passage showed that PTHs were selectively infected with defined HCV quasispecies, and new quasispecies emerged in PTHs after passage. CONCLUSION: The present study strongly indicates that PTHs could be infected by HCV and support HCV replication in vitro. Our results would be helpful for the establishment of a tupaia model of HCV infection. PMID- 16313721 TI - [A method of HPRE synthesis via transcription by T7 RNA polymerase in vitro]. AB - OBJECTIVE: To synthesize highly pure HBV post-transcriptional regulatory element (HPRE) via transcription in vitro by T7 RNA polymerase. METHODS: HPRE gene was amplified by PCR from a template containing HBV complete genomic DNA and cloned into plasmid pGEM-11zf. The cloned DNA sequence was transcribed by T7 RNA polymerase. RESULTS: The construction of HPRE gene recombinant plasmid and production of HPRE via transcription in vitro was successful. CONCLUSION: In vitro transcription by T7 RNA polymerase can be used to synthesize highly pure HPRE. PMID- 16313722 TI - [Genomic analysis of a familial clustering of chronic hepatitis B patients]. AB - OBJECTIVE: To investigate the peripheral blood monocyte (PBMC) gene expression profile in a familial clustering of patients with chronic hepatitis B (CHB). METHODS: cRNA prepared from PBMC in a family with 5 CHB patients and 4 normal controls was hybridized to high-density oligouncleotide arrays (HG-U133A 2.0 Human GeneChips, Affymetrix), which interrogate the expression of approximately 22,000 human ESTs. Primary image obtained from scanning was analysed with a DNT software package. Real-time PCR was employed to confirm the gene chip results. RESULTS: 55 genes out of 22,000 ESTs were identified differently. Among the 55 genes 14 showed increased expression and 41 showed decreased expression in the familial clustering CHB patients compared with those in normal controls. Most of the genes (57%) were involved in immunity, inflammation, apoptosis, signaling transduction, and cell cycle. CONCLUSION: These results suggest that the hosts with this broad range of gene expression alterations are susceptible to hepatic B infection. PMID- 16313723 TI - [Aldosterone stimulates alpha1-(1) procollagen mRNA expression in HSC via activation of ERK1/2 and AP-1]. AB - OBJECTIVE: It has been known that the intrahepatic renin-angiotensin-aldosterone system (RAAS) plays a key role in the fibrogenesis in livers. Aldosterone (Aldo), the principal effector molecule of the RAAS, exerts local effects on cell growth and fibrogenesis. However, the signal transduction mechanisms underlying the effects of Aldo on hepatic fibrogenesis remain to be fully elucidated. The present study aims to investigate the signal transduction mechanism underlying the effects of Aldo on the signal passageway of active protein-1 (AP-1). METHODS: In vitro, HSCs-T6 cell line was treated with Aldo for 10 min, 30 min, 60 min, 120 min and 180 min, and protein expression of Phospho-P42/44 was detected by Western blot. In addition, HSCs-T6 cell line was preincubated for 60 min or not with U0126 (an inhibitor of the MAPK/ERK kinase), and also with antioxidant-N acetylcysteine (NAC) prior to exposure to Aldo for the indicated times. Protein expression of Phospho-P42/44 was measured by Western blot. DNA biding activity of AP-1 was analyzed by electrophoretic gel mobility shift assay (EMSA). By means of RT-PCR, expression of alpha1(1) procollagen mRNA was detected. RESULTS: Aldo stimulated HSC via extracellular signal-regulated kinase (ERK1/2) pathway. Time course experiments showed that Aldo induced Phospho-P42/44 expression, which was abrogated by U0126, reaching a maximum at 10 minutes, and then declined progressively. NAC inhibited the Phospho-P42/44 expression. EMSA showed that stimulation of HSC by Aldo markedly increased AP-1 DNA binding activity. U0126 markedly reduced AP-1 DNA binding activity induced by Aldo; NAC partly decreased AP-1 activity induced by Aldo. Aldo up-regulated expression of alpha1(1) procollagen mRNA, which was attenuated by U0126 and NAC. CONCLUSION: Stimulation of HSC by Aldo results in activation of AP-1 via ERK1/2 pathway, leading to up regulation of AP-1 target gene alpha1(1) procollagen mRNA expression. PMID- 16313724 TI - [An experimental study of the inhibitory effects on the activation of endotoxin induced Kupffer cells through short hairpin RNA targeting interleukin-1 receptor associated kinase-4 gene]. AB - OBJECTIVE: To explore the inhibitory effects on the activation of endotoxin induced Kupffer cells (KCs) through short hairpin RNA (shRNA) targeting interleukin-1 receptor associated kinase-4 (IRAK-4) gene. METHODS: Two effective transfection shRNA plasmid (pSIIRAK-4-A, pSIIRAK-4-B) and one invalidated plasmids (pSIIRAK-4-C) targeting IRAK-4 gene were constructed. The isolated mouse KCs were divided into three groups: the normal control group, the RNAi control group (pSIIRAK-4-C) and the RNAi effective group (pSIIRAK-4-A, pSIIRAK-4-B). Then KCs were stimulated with 0.1 microg/ml lipopolysaccharide (LPS) after 24 h transfection with the constructed plasmid. The expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blot at 6 h after LPS stimulation, and the activities of NF-kappaB in KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h. RESULTS: The level of IRAK-4, the activities of NF-kappaB and the TNF-alpha level in the RNAi effective group were evidently lower than those in normal and RNAi control groups (P < 0.01) at 1 h, 3 h, and 6 h. Especially, the pSIIRAK-4-A group in which the changes of the above indices were of no difference (P > 0.05), had better inhibited effects than that of the pSIIRAK-4-B group (P < 0.01). CONCLUSION: The shRNA targeting IRAK-4 gene could effectively inhibit the activation of endotoxin-induced KCs. PMID- 16313725 TI - [The dynamic expression of Thy1.1 positive hepatic oval cells during the shaping and reducing processes of liver cirrhosis induced by dimethylnitrosamine in rats]. AB - OBJECTIVES: To study the dynamic change of hepatic oval cells (HOC) in the process of rat liver cirrhosis formation induced by dimethylnitrosamine (DMN), and to explore its pathophysiology significance. METHODS: A rat cirrhosis model was established by using DMN. Microscopical and electronmicroscopical changes of HOC were examined. Thy1.1 was detected by immunohistochemical method at different times. The ratio of HOC was checked using image pattern analysis and Western blot. The number of HOC was counted microscopically. RESULTS: At the 4th week after DMN administration, the liver fibrosis was at its peak, with false lobules formation combined with large areas of hemorrhage and necrosis. The fibrosis started to minimize at the 6th week, and also the inflammatory changes at the 8th week. Thy1.1 positive stained cells dispersed at the 2nd week; increased at the 4th week around fiber septa; reached its peak at the 6th week, then decreased at the 8th week. The results of image pattern analysis, cell counting under light microscope and Western blot were constant, with the highest cell numbers at the 6th week, and dropped at the 8th week. The ultrastructure of HOC was characterized by their small sized, oval nuclei, and higher nucleus/plasma ratio. CONCLUSION: During the formation and reduction of rat cirrhosis caused by DMN, Thy1.1 stained HOC showed notable dynamic change, which may play an important role in the cirrhotic process. PMID- 16313726 TI - [A case of subacute liver failure resulted from bee venom]. PMID- 16313727 TI - [Effects of glycyrrhizin on TGFbeta1 stimulated hepatic stellate cell signaling transduction]. AB - OBJECTIVES: To investigate the role of glycyrrhizin on TGFbeta1 stimulated signaling transduction in rat hepatic stellate cells (HSCs). METHODS: The mice HSCs were isolated and cultured with or without glycyrrhizin (1 micromol/L-1000 micromol/L) in vitro after TGFbeta1 stimulation. The mRNA level of Smad2, 3, 7 were measured with RT-PCR; protein expression level of Smad2, 3, 7 and collagen I, III were analyzed with Western blot. RESULTS: TGFbeta1 increased the mRNA level and protein expression of Smad2, 3, 7 in HSC; it also increased protein expression of collagen I and III. 1 micromol/L-1000 micromol/L glycyrrhizin decreased the mRNA level and protein expression of Smad2, 3, 7; it also inhibited protein expression of collagen I and III gradually. CONCLUSION: Interventing the TGFbeta signaling pathway and decreasing the synthesis of collagen, might be involved in the anti-fibrosis mechanism of glycyrrhizin. PMID- 16313728 TI - [Screening hepatocellular carcinoma autoantibodies by serological proteome analysis]. AB - OBJECTIVE: To screen hepatocellular carcinoma (HCC) autoantibodies as diagnostic biomarkers or therapy targets by serologic proteome analysis (SERPA). METHODS: Total proteins extracted from human HCC cell line HCCLM3 were separated by two dimensional electrophoresis (2-DE) and then transferred onto PVDF membranes, which were subsequently incubated with sera from HCC, hepatitis B virus (HBV) infected patients or healthy volunteers. All immuno-reactive protein spots on blot films were matched to those on 2-DE gel maps by image analysis and identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS/MS). RESULTS: 2-DE gel maps of HCCLM3 and corresponding blot films of good quality and reproducibility were established. The number of spots on HCCLM3 2-DE reference gel totaled 603 and those on HCC, HBV and healthy sera blotted films were 70.75+/-24.25, 68.5+/-23.44 and 41.38+/ 15.05, respectively. Blot films of HCC and HBV groups had more spots than those of the healthy group (P < 0.05) while no significance was found between films of HCC and HBV groups. By identification, those HCC autoantibodies could be classified as nuclear proteins, cytoskeleton proteins, heat shock proteins and metabolic enzymes. CONCLUSION: Serological proteome analysis is a high throughput technique for screening tumor autoantibodies. Those newly identified HCC associated tumor antigens and corresponding autoantibodies can be used in the early diagnosis or immuno-therapy of HCC. PMID- 16313729 TI - [Preliminary study on the effects of an artificial liver support system in the treatment of medicamentous liver insufficiency]. AB - OBJECTIVES: To study the effectiveness of an artificial liver support system. METHODS: Thirty-two patients with medicamentous liver insufficiency were treated with an artificial liver support system in addition to the routine medicinal therapy. Thirty patients treated with routine medicinal therapy only served as controls. RESULTS: The clinical symptoms (e.g. hepatic encephalopathy) and the laboratory indices (serum total bilirubin and prothrombin time) of the treatment group patients were obviously improved compared with those of the control group patients (P < 0.05). The cure rate and hospitalization days were 90.6% (26/32) and 47 days respectively in the treatment group, and 43.3% (13/30) and 72 days in the control group (P < 0.05). CONCLUSION: Using an artificial liver support system combined with routine medicinal therapy is more effective than using medication alone. PMID- 16313730 TI - [The function of immuno-modulating on CD1a dentritic cells by IFN-alpha in chronic hepatitis B patients in vivo]. PMID- 16313731 TI - [A study on stem cell mobilization after partial liver transplantation in rats]. AB - OBJECTIVE: To explore the role of stem cell mobilization on regeneration of partially grafted livers. METHODS: Rats models with cross-sex 50% PLTx (partial liver transplantation) were established. The rats were divided into three groups: PLTx, WLTx (whole liver transplantation) and sham operation groups. Bone marrow and liver samples were collected on days 1, 3, 5, 7 postoperatively (each n = 6). The quantitative variations of the cells with stem cell markers in the bone marrow, including beta2m-/Thy-1.1+, CD45+/CD34+, Flt2/3+ and c-kit+ markers, were detected using flow cytometry. Sry gene positive cells in donor livers were detected by fluorescent in situ hybridization (FISH), and the expressions of CD34, c-kit and Thy-1.1 were detected by immunohistochemistry technique. RESULTS: Compared with the WLTx and sham operation groups, beta2m-/Thy-1.1+, CD45+/CD34+ cells in bone marrows in the PLTx group increased on the first postoperative day and decreased on the following days. The CD34, c-kit and Thy-1.1 positive cells detected in portal tract areas peaked during the 3-5 postoperative days. CD34+/CD45+ positive cells could be detected. The expressions of CD34, c-kit and Thy-1.1 positive cells were rare in the WLTx and sham operation groups. Sry+ cells could be detected in portal tract areas and few Sry+/CD34+ and Sry+/Thy 1.1+cells were detected. CONCLUSION: In the PLTx group, the stem cells in the bone marrow were mobilized and stem cells in the liver were activated. PMID- 16313732 TI - [Progress in clinical application of artificial liver support systems]. PMID- 16313733 TI - [Progress of artificial liver support system technology]. PMID- 16313734 TI - [Prospects for cell sources of hepatocyte transplantation and bioartificial liver]. PMID- 16313735 TI - [Relationship between the distribution of hepatitis B virus genotypes and clinical types of the disease in Han nationality hepatitis B patients in Xinjiang]. PMID- 16313736 TI - [An investigation of the relationship between heavy drinking and alcoholic fatty liver in the Xinjiang minority ethnic group]. PMID- 16313737 TI - [Sexual differences in hepatic ischemic reperfusion injury in rats are related to endothelial cell nitric oxide synthase-derived nitric oxide]. PMID- 16313738 TI - [Clinical characteristics of 22 autoimmune hepatitis patients]. PMID- 16313739 TI - [Protective effects of rosiglitazone on hepatotoxicity in hepatocytes cultured with cyclosporine A]. PMID- 16313740 TI - [Effect of tumor necrosis factor-alpha on blood-brain barrier permeability in mice with acute liver necrosis]. PMID- 16313741 TI - [The prevention and management of cerebrovascular complications in nine cases after orthotopic liver transplantation]. PMID- 16313742 TI - [The value of serum phosphorus level analysis in assessing clinical characteristics of hepatitis B patients]. PMID- 16313743 TI - [Two kinds of cytokines induce multipotent adult progenitor cells from human bone marrow differentiating into hepatocyte-like cells in vitro]. PMID- 16313744 TI - [CD45RA expression changes and their significance in hepatitis and in hepatic cancer patients]. PMID- 16313745 TI - [Early diagnostic value of abnormal expression of insulin-like growth factor (IGF)-II and IGF-II-mRNA in hepatocellular carcinoma patients]. PMID- 16313746 TI - [The effects of plasma from patients with severe hepatitis on the growth and function of porcine hepatocytes]. PMID- 16313747 TI - [Effect of phosphoinositide 3-kinase/Akt-significance on the secretion function of rat SEC after partial hepatectomy]. PMID- 16313748 TI - [Occult hepatitis B virus infection]. PMID- 16313749 TI - [Development of prognostic models of liver diseases]. PMID- 16313750 TI - [Methods for diagnosis of minimal hepatic encephalopathy and their evaluation]. PMID- 16313751 TI - [Outcome of 1,355 consecutive transabdominal chorionic villus samplings in 1,351 patients]. AB - BACKGROUND: The true risk of chronic villus sampling (CVS) is poorly defined. The objective of this study was to review the clinical outcome of transabdominal CVS performed in a university teaching unit, with an emphasis on the complication rate. METHODS: A comprehensive audit database was maintained for 1,351 pregnant women, including 17 sets of twin pregnancies, who had a CVS. Details and outcome of all CVSs made in the unit between May 1996 and May 2004 were reviewed. All CVSs were performed by one of 5 operators using the identical techniques. RESULTS: All procedures were performed transabdominally. A total of 1,355 CVSs were performed because there were 4 dichorionic twin pregnancies which required 2 punctures. The mean gestation at CVS was (11.8 +/- 0.7) weeks, and 97.3% of the procedures were performed between 11 and 13 completed weeks. The majority (96.2%) required only 1 puncture to achieve correct needle placement. The procedure failed to obtain an adequate sample in 4 subjects (0.30%). A total of 1,351 chromosomal studies were requested and there was 1 case (0.07%) of culture failure. The results of chromosomal studies were available within 14 days in 36.7% of the cases and within 21 days in 94.0%. Overall, 77 chromosomal abnormalities (5.7%) and 5 cases of thalassemia major were detected. Pregnancy outcome was unknown in only 13 singleton subjects (0.96%). In the remaining 1,355 fetuses, there were 76 pregnancy terminations (5.56%), 10 fetal losses with obvious obstetric causes (0.73%), and 21 potentially procedure-related fetal losses (1.54%). In the last group, the majority had one or more co-existing obstetric complications. The background fetal loss rate for pregnancies at similar gestational age in the unit was about 0.8%. Therefore, the procedure related fetal loss rate was estimated to be at the maximum of 0.74%. CONCLUSIONS: In experienced hands, first trimester transabdominal CVS is an accurate and safe invasive prenatal diagnostic procedure. It should be one of the treatment options available to pregnant women who require prenatal genetic diagnosis. PMID- 16313752 TI - [Intraperitoneal hemorrhage during and after percutaneous radiofrequency ablation of hepatic tumors: reasons and management]. AB - BACKGROUND: Intraperitoneal hemorrhage is one of the most common complications of radiofrequency (RF) ablation of hepatic tumors. This study was designed to investigate the reason and management of intraperitoneal hemorrhage occurred during or after percutaneous RF ablation of hepatic tumors. METHODS: Three hundred and fifty-six patients with hepatic tumors have been treated at 592 procedures of ultrasound guided RF ablation. Intraperitoneal hemorrhage occurred in 5 patients (0.8%). The reasons and management of intraperitoneal hemorrhage in these 5 cases were retrospectively analyzed. RESULTS: Two patients with liver metastasis and one hepatocellular carcinoma (HCC) patient suffered from hemorrhage during the RF treatment. Two patients with recurrent HCC after surgery developed hemorrhage 20 minutes or 4 hours after RF treatment. One case of hemorrhage was due to the inappropriate electrode positioning induced liver laceration while treating a 1 cm liver metastasis near the liver capsule. One was due to the injury of a small vessel by the RF needle in another liver metastasis patient. Three cases were due to tumor rupture with two cases induced by cough or position change after treating large protruding HCC lesions. Four (80%) of the 5 cases of hemorrhage were rapidly identified by ultrasound. The causes and sites of bleeding during the RF treatment in three cases were confirmed through ultrasound, which were successfully treated using RF coagulation to achieve hemostasis of the bleeding site. Two patients with post-ablation hemorrhage recovered in one hour and 24 hours, respectively after given blood transfusion and other conservative measures. No surgical intervention was required. Two patients died of wide spread metastasis 23 - 36 months afterwards and the other three patients have lived for 18 - 25 months to date. CONCLUSIONS: It is important to perform close monitoring during and after RF ablation in order to identify intraperitoneal hemorrhage in time. RF ablation of the bleeding sites was a simple and effective management when the bleeding site could be confirmed by ultrasound. The hemorrhage due to the rupture of large and protruding liver tumors could be serious and should be considered as contraindication for RF treatment. PMID- 16313753 TI - [Anti-tumor effect of thalidomide and paclitaxel on hepatocellular carcinoma in nude mice]. AB - BACKGROUND: Thalidomide is reviving for its antiangiogenic effect on corneal neovascularization models. Recently, it has been employed in tumor research in several types of solid carcinomas. However, its effect on hepatocellular carcinoma (HCC) has not yet been clarified. METHODS: A total of 48 nude mice bearing human HCC with a high metastatic potential were randomly divided into 4 groups. Thalidomide (200 mg/kg), paclitaxel (13 mg/kg), or their combination, which was dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension, was intraperitoneally injected in each group since the second day of the establishment of animal model. The group simply administered with 0.5% CMC was set as placebo-control. The mice were sacrificed on the 30th day, for the measurement of tumor size, weight and metastasis in the lungs. The levels of CD34 and endothelial growth factor (VEGF) mRNA in tumor tissues were detected by immunohistochemistry and semiquantitative RT-PCR, respectively, and microvessel density (MVD) was evaluated. RESULTS: No statistical difference was found in tumor weight and volume between the thalidomide group and control (P>0.05). Paclitaxel showed a growth-inhibiting effect on tumors (P<0.05). The value of MVD and VEGF mRNA and metastases to the lungs in each group were lower than those in the placebo-control group (P<0.05); such difference in the combination group was statistically significant (P<0.05). CONCLUSIONS: Paclitaxel, but not thalidomide, has significant growth inhibitory effect on tumors, but both significantly inhibit angiogenesis and metastasis of human HCC in nude mice, such effects of paclitaxel can be amplified by thalidomide. PMID- 16313754 TI - [Mechanism of reduction of albumin expression induced by lipopolysaccharide in rat hepatocytes]. AB - BACKGROUND: The severity of hypoalbuminemia has been shown to be related to morbidity and mortality in some critical illnesses, illustrating the need for better understanding of molecular mechanism of hypoalbuminemia. Lipopolysaccharide (LPS) is a key mediator inducing hypoalbuminemia in sepsis and septic shock. The present study was designed to identify if the reduction of albumin expression is directly induced by LPS and modulated by activated extracellular signal-regulated protein kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) in rat hepatocytes. METHODS: Primary rat hepatocytes were divided into five groups. In two of them, hepatocytes were treated with normal saline or 1 microg/ml LPS, then albumin mRNA expression was observed at 0, 2, 8, 12 and 24 hours after treatment. In another group, hepatocytes were pretreated with 100, 40 or 20 micromol/L of cycloheximide (CHX, an inhibitor of protein synthesis) for 30 minutes followed by 1 microg/ml LPS for 24 hours. Then the RNA was extracted from the cells for RT-PCR to detect the expression of albumin. The other two groups were administered 1 micromol/L, 10 micromol/L and 50 micromol/L of SB203580 (p38 MAPK inhibitor) or PD98059 (ERK inhibitor) 30 minutes prior to 1 microg/ml LPS treatment. After 24 hours of LPS treatment, the supernatant was collected and assayed for albumin concentrations. Data were analyzed by one-way analysis of variance, followed by the Newman-Keul test; a P<0.05 was considered significant. RESULTS: There was no marked change in albumin mRNA expression in the control group during 24-hours treatment with normal saline. The reduction did not occur until 24 hours after LPS treatment, and albumin mRNA decreased by 30% approximately compared to the control group at 24 hours (0.587 vs 0.832, P=0.007). CHX could inhibit the decline of albumin mRNA induced by LPS and the effect was correlated with the dose of CHX. The ERK inhibitor PD98059 caused a significant increase in LPS-induced albumin production at the three concentrations (119.7, 111.4 and 80.0 ng/ml vs 44.4 ng/ml, P=0.0013, 0.0025 and 0.009, respectively), whereas SB203580 obviously blocked albumin reduction in LPS treated cells at the concentrations of 10 and 50 micromol/L (87.5 and 93.6 ng/ml vs 44.4 ng/ml, P=0.0076 and 0.0049, respectively). CONCLUSIONS: LPS can induce the reduction of albumin expression by new synthesized proteins indirectly, and the process may be related to the signal proteins of ERK and p38 kinase. The ERK and p38 kinase are critical signaling pathways in LPS-induced hypoalbuminemia which is worthwhile to understand in studying the molecular mechanism of hypoalbuminemia in sepsis and septic shock. PMID- 16313755 TI - [Effects of secretive bone morphogenetic protein 2 induced by gene transfection on the biological changes of NIH3T3 cells]. AB - BACKGROUND: Bone morphogenetic proteins (BMPs), which belong to the transforming growth factor beta superfamily, are powerful regulators of cartilage and bone formation. This study investigated the biological changes of NIH3T3 cells incubated with secretive BMP2 that was induced by gene transfection through transwell. METHODS: Eukaryonic expression vector (pcDNA3.1-B2) was transfered into NIH3T3 cells with Sofast, a positive compound transfection agent. The positive cell clones were selected with G418. The cytoplasmic and extracellular expressions of BMP2 were determined by immunohistochemical stain and enzyme linked immunosorbent assay. NIH3T3 cells were co-cultured with hBMP2 gene transfecting cells through transwell, and the ultrastructure, alkaline phosphatase activity and the expression of osteocalcin (the marker of osteogenetic differentiation) changes were observed. RESULTS: There were cytoplasmic and extracellular expressions of BMP2 in transfecting NIH3T3 cells. The ultrastructural changes, the high activity of alkaline phosphatase and the positive stain of osteocalcin suggested the osteogenetic differentiation tendency of NIH3T3 cells co-cultured with transfecting NIH3T3 cells. CONCLUSION: Secretive BMP2 that is induced by gene transfection could promote the osteogenetic differentiation of fibroblast cells. PMID- 16313756 TI - [Differential expression of Bmp2, Bmp4 and Bmp3 in embryonic development of mouse anterior and posterior palate]. AB - BACKGROUND: The palate is differently regulated and developed along the anterior posterior axis. The Bmp signal pathway plays a crucial role in palatogenesis. Conditioned-inactivation of Bmp type I receptor Alk2 or Alk3 in the neural crest or craniofacial region leads to palatal cleft in mice. However, how different Bmp members are involved in palatogenesis remains to be elucidated. In the present study, mRNA expression patterns of Bmp2, Bmp3 and Bmp4 in the developing anterior and posterior palates were examined and compared, focusing on the fusion stage. METHODS: To detect the expression of Bmp mRNA, antisense riboprobes were synthesized by in vitro transcription. Radioactive in situ hybridization was performed on sagital and coronal sections of mice head from E13 to E18. RESULTS: The expression of these Bmps were developmentally regulated in the anterior and posterior palates prior to, during and after palatal fusion. During palatal fusion, Bmp4 expression shifted from the anterior to the posterior palate, Bmp2 was highly expressed in both the anterior and posterior palates in this process, whereas Bmp3 was only localized in the posterior palate. They showed generally non-overlapping pattern in their expression domains. Thereafter, their expression was detected in both the anterior and posterior palates regulating osteogenesis and myogenesis respectively. CONCLUSIONS: Bmp signalling is involved in palatogenesis in multiple stages and has multiple roles in regulating anterior and posterior palatal development. Disturbances of Bmp signalling during palatogenesis might be a possible mechanism of cleft palate. PMID- 16313757 TI - [Mechanism linking atherosclerosis and type 2 diabetes: increased expression of scavenger receptor CD36 in monocytes]. AB - BACKGROUND: We investigated the pathogenesis of atherosclerosis in diabetes, and detected the expression of scavenger receptor CD36 in monocytes in patients with type 2 diabetes. METHODS: According to the criteria by WHO, diabetic patients were classified into two groups: well controlled diabetic patients (WCP) and poorly controlled diabetic patients (PCP). The expression of CD36 protein and mRNA were evaluated by flow cytometry and reversal transcription polymerase chain reaction (RT-PCR). Plasma levels of accumulation of oxidized LDL (oxLDL) were directly measured by sandwich enzyme-linked immunosorbent assay (ELISA) method. RESULTS: Flow cytometry and RT-PCR showed that the mean fluorescence intensity (MFI) of CD36 in monocyte and CD36 mRNA were significantly higher in the PCP and WCP in comparison with healthy controls (P<0.01). CD36 MFI and mRNA in the PCP were increased by 78% and 36% compared to the WCP. In both groups, CD36 MFI and mRNA were significantly higher in patients with diabetic atherosclerosis in comparison with those without diabetic atherosclerosis (P<0.05). No significant difference was found in CD14 expression between the groups (P>0.05). The concentrations of plasma oxLDL were higher in the PCP group compared to WCP and control group (P<0.05), whereas oxLDL average values did not differ significantly between WCP and control groups (P>0.05). In the WCP and PCP groups, oxLDL levels were higher in patients with diabetic atherosclerosis than those without diabetic atherosclerosis (P<0.05). CONCLUSIONS: The increased expression of scavenger receptor CD36 may be one of the mechanism of accelerated atherosclerosis in diabetic. The poorly controlled diabetes patients are at higher risk for the vascular complications than the well controlled diabetic patients. PMID- 16313758 TI - [Mechanism of the protective effects of noninvasive limbs preconditioning on myocardial ischemia-reperfusion injury]. AB - BACKGROUND: This study aimed at assessing the effect of noninvasive limb preconditioning on myocardial infarct size, and determining whether nitric oxide and neurogenic pathway play an important role in the mechanism of acute remote ischemic preconditioning (IPC). METHODS: Forty Wistar rats were randomly divided into four experimental groups. In Group I, the rats underwent 30-minute occlusion of the left anterior descending coronary artery, and 120-minute reperfusion. In Group PL, the rats underwent four cycles of 5-minute occlusion and reperfusion of both hind limbs using a tourniquet before the experiment was continued as in Group I. In Group PL-N and Group PL-H, we administered L-nitro-arginine methyl ester (L-NAME) 10 mg/kg or hexamethonium chloride 20 mg/kg intravenously, 10 minutes before IPC. Infarct size as a percentage of the area at risk was determined by triphenyltetrazolium chloride staining. RESULTS: There were no statistically significant differences in mean arterial pressure and heart rate among these groups at any time point during the experiment (P>0.05). The myocardial infarct size (IS) was decreased significantly in Group PL and Group PL H compared with Group I, and the IS/AAR was 34.5%+/-7.6%, 35.9%+/-8.6% and 58.5%+/-8.5%, respectively (P< 0.05). The IS/AAR was 49.1%+/- 6.5% in Group PL-N, and there was no significant difference compared with Group I (P>0.05). CONCLUSIONS: Noninvasive limb IPC is effective in protecting the myocardium from ischemia reperfusion injury. Nitric oxide plays an important role in the mechanism of acute remote IPC, in which the neurogenic pathway is not involved. PMID- 16313759 TI - [Mesenchymal stem cells transduced by PLEGFP-N1 retroviral vector maintain their biological features and differentiation]. AB - BACKGROUND: Enhanced green fluorescent protein (EGFP) has been an important reporter gene for gene therapy. Human mesenchymal stem cells (hMSCs) are ideal target cells in cell transplantation and tissue engineering. We investigated their biological characteristics and differentiation mediated by PLEGFP-N1 retroviral transduction. METHODS: hMSCs were isolated from human bone marrow by density gradient fractionation and adherence to plastic flasks. Individual colonies were selected and cultured in tissue dishes. Packaging cells PT67 were transfected by PLEGFP-N1 retroviral vector, and hMSCs were transduced by viral supernatant infection. Meanwhile, hMSCs-EGFP were identified by immune phenotypes and whether it could differentiate into osteoblasts or adipocytes under conditioned media was investigated. RESULTS: The rate of stably transduced hMSCs EGFP was up to 96% after being screened by G418. hMSCs-EGFP exhibited fibroblast like morphological features. Flow cytometric analyses showed that hMSCs-EGFP were positive for CD73, CD105, CD166, CD90 and CD44, but negative for CD34 and CD45. In addition, it could functionally be induced into osteocytes or adipocytes under conditioned media. These biological features of hMSCs-EGFP were consistent with those of hMSCs. CONCLUSIONS: hMSCs transduced by PLEGFP-N1 retroviral vector can be used in vivo securely because they can maintain their biological characteristics and differentiation. It is a simple and reliable way to trace the changes of hMSCs in vivo by EGFP during cell transplantation and gene therapy. PMID- 16313760 TI - [KCNE3 R53H substitution in familial atrial fibrillation]. PMID- 16313761 TI - [Detection and evaluation of the mutations of embB gene in ethambutol-susceptible and resistant Mycobacterium tuberculosis isolates from China]. PMID- 16313762 TI - [Glucocorticoids upregulate transepithelial electrical resistance and expression of tight junction-related protein in human trabecular meshwork cells]. PMID- 16313763 TI - [DK crush technique: modified treatment of bifurcation lesions in coronary artery]. PMID- 16313764 TI - [Congenital aortic arch anomalies: diagnosis using contrast enhanced magnetic resonance angiography]. PMID- 16313765 TI - [Prognostic factors of fulminant hepatitis in pregnancy]. PMID- 16313766 TI - [Intranodal palisaded myofibroblastoma and differential diagnosis: a case report]. PMID- 16313770 TI - [Carrier rate of Streptococcus pneumoniae and susceptibility thereof to antimicrobial drugs among children in China: a surveillance study in Beijing, Shanghai, and Guangzhou 2000-2002]. AB - OBJECTIVE: To investigate the carrier rate of Streptococcus pneumoniae and the susceptibility of S. pneumoniae to 10 different antimicrobial drugs among children in China. METHODS: Nasopharageal swab specimens were collected from randomly selected 3578 out-patients with upper respiratory infection aged 1 month to 5 years in Beijing, Shanghai, and Guangzhou 2000-2002 so as to isolate S. pneumoniae. The susceptibility of these strains to erythromycin and beta-lactam antibiotics (penicillin, amoxicillin/clavulanic acid, cefaclor, cefuroxime and ceftriaxone) was determinated by E-test MIC method, and the susceptibility to chloramphenicol, tetracycline, sulfamethoxazole/trimethoprim and ciprofloxacin was determinated by disk diffusion method. RESULTS: A total of 892 strains of S. pneumoniae were isolate from the 3578 children with a carrier rate of 24.9% (9.0% 36.0%). 634 isolates of S. pneumoniae were tested to examine their susceptibility to chloramphenicol, 199 to ciprofloxacin, and 887 to the other 8 antimicrobial drugs. The insusceptibility rate of S. pneumoniae to penicillin was 39.9%, and the resistance rate was 6.4%. The resistance rate to cefaclor was 20.4%. The insusceptibility rate to cefuroxime was 19.0%. The susceptibility rate to amoxicillin/clavulanic acid was 98.5% and that to ceftriaxone was 97.2%. More than 80% of the isolates were resistant to erythromycin, tetracycline, and sulphamethoxazole/trimethoprim. Chloramphenicol resistance was less common (32.3%) than the resistance to other non-beta-lactam antibiotics, except for ciprofloxacin to which the resistance rate was 2.0%. The national insusceptibility rates for beta-lactam antibiotics did not vary significantly during the three years with the widest varying range of 5.1% for penicillin (38.1%-43.2%). Each of the three cities had their different varying patterns of antimicrobial susceptibility, especially for beta-lactam antibiotics. Multi-drug resistance was common (88.7%). Strains of S. pneumoniae insusceptible to penicillin were significantly more common than those susceptible to penicillin (99.1% vs. 78.8%, chi(2) = 50.36, P < 0.001), and strains of S. pneumoniae insusceptible to erythromycin were significantly more common than those susceptible to erythromycin too (95.8% vs. 17.1%, chi(2) = 360.26, P < 0.001). CONCLUSION: Antimicrobial resistant S. pneumoniae has already become a serious problem in China. Ongoing surveillance study on the antimicrobial resistance of S. pneumoniae is necessary for appropriate antimicrobial use in clinical work and modification of medical strategies for people's health. PMID- 16313771 TI - [Association of clinical features with different hemodynamic patterns in head-up tilt test in children with unexplained syncope]. AB - OBJECTIVE: To explore the different hemodynamic patterns during the course of head-up tilt tests in children with unexplained syncope and the association of clinical features with different hemodynamic patterns in head-up tilt test in children with unexplained syncope. METHODS: 100 pediatric patients with unexplained syncope, aged 11 +/- 2 (6-16), 36 male and 64 female, with the mean course of 10 +/- 16 months (2 d-5 y), underwent head-up tilt tests or head-up tilt tests potentiated with nitroglycerine under quiet circumstance. Blood pressure and heart rate were monitored simultaneously. According to their different hemodynamic patterns, they were divided into vasovagal response pattern, postural orthostatic tachycardia syndrome (POTS) response pattern, orthostatic hypotension (OH) response pattern and normal response pattern. The vasovagal response was divided into vasodepressor, cardioinhibitory and mixed patterns. The distribution and different clinical features of different response patterns in the unexplained syncope were also studied. RESULTS: Fifty (50%) of the 100 children with unexplained syncope displayed the hemodynamic pattern of vasovagal response, among which 31 (31%) displayed the pattern of vasodepressor response, 7 (7%) cardioinhibitory response, and 12 (12%) mixed response. Thirty three patients (33%) displayed POTS response, 2 (2%) OH response, and 15 (15%) the normal hemodynamic response. Patterns of dysautonomic response and chronotropic incompetence were not observed in these children with unexplained syncope. The age of the children with normal response during HUT was 10 +/- 3 years, significantly younger than that of the children with vasovagal response and POTS response (12 +/- 2 and 12 +/- 2, both P < 0.01). There were no differences in sex ratio and duration of syncope among the vasovagal response, POTS and normal response. But the syncopal spells in the children with POTS response was less frequent and the baseline heart rate of the children with POTS response was 81 +/- 7, significantly faster than that of the children with vasovagal response and normal response (71 +/- 9 and 74 +/- 7, both P < 0.01). There was no significant difference in the baseline blood pressure among the children with vasovagal response, POTS and normal response. There were also no significant differences in the age, sex ratio, and duration of syncope, number of syncopal spells, baseline heart rate, and baseline blood pressure among the children with vasodepressor response, cardioinhibitory response and mixed response. CONCLUSION: There are different hemodynamic response patterns in head up tilt testing in children with unexplained syncope, and there was some association between hemodynamic response patterns and their clinical features. PMID- 16313772 TI - [Risk factors of familial gallstone disease: study of 135 pedigrees]. AB - OBJECTIVE: To explore the role of both inheritable and epidemiological factors in the pathogenesis of familial gallstone disease in pedigrees. METHODS: 135 pedigrees, with 695 members aged > or = 18 (18 - 83, with a mean of 50 +/- 14), 282 males and 413 females, including 370 patients, at least one patient existing in every generation, were investigated. Inquiry was carried out to collect the personal and medical history. Physical examination and ultrasonography were conducted. Blood samples were collected to detect the total cholesterol (TCh), triglyceride (TG), high-density lipoprotein cholesterol (HDL-Ch), and apolipoprotein (Apo) A1 and ApoB. The concentration of low-density lipoprotein cholesterol (LDL-Ch) was calculated by the formula: Tch-TG/5 + HDL-Ch. The inheritable characteristics and heritability were analyzed. RESULTS: The prevalence of gallstone disease was 53.24%, 56.66% among the females, significantly higher than that among the males (48.23%, P = 0.03). The heritability in the first-degree relatives was 138% +/- 7%, and the concordance rate of monozygotic twins was 100%. The significant risk factors were female gender (P = 0.03), body mass index (P < 0.01) and diet (P < 0.008). The history of hypertension, hyperlipidemia, and diabetes were correlated to gallstone disease (P = 0.008, < 0.008, and 0.03 respectively). Serum HDL-Ch and ApoA1 concentrations were lower in the stone group than in the non-stone group (P = 0.003 and 0.005 respectively). There were no significant differences in TCh, TG, LDL-Ch, and ApoB between the 2 groups. CONCLUSION: Genetic factors play a major role in the pathogenesis of familial gallstone disease, characterized by autosomal dominant inheritance. Female gender, obesity, diet, hypertension, hyperlipidemia, diabetes may be the risk factors of gallstone disease. Dyslipidemia is a characteristic of gallstone disease. PMID- 16313774 TI - [Importance of colonoscopy combined with biopsy in the diagnosis of colonic Crohn's disease]. AB - OBJECTIVE: An attempt was made to provide a better insight into endoscopic and histological features and/or problems encountered when establishing a diagnosis of colonic Crohn's disease (Crohn's colitis). METHODS: As presented in our 27 cases with Crohn's colitis, the endoscopic findings and histological changes of biopsy specimens were summarized. As collated with correspondent results of biopsy and surgical specimens, the diagnostic accuracy of endoscopy was evaluated. RESULTS: twenty-six involvements of the large intestine (combined with other sites of the intestine) was found (96.3%). However, involvement limited to the colon alone was seen in only 4 cases (14.8%). Endoscopically, overlapped lesions with multistaged-segmental distributed and multi-sited diverse patterns (destructive and proliferative/regenerative) of inflammatory changes were observed. Endoscopic accuracy was 66.7%. The characteristic features of mucosal biopsy include focal distribution of the lesions, a thickened and edematous submucosa, deep fissuring ulcers, lymphoid aggregate, and hyperplasia, fibrosis and granulomas (detected in 30% of the group), etc. CONCLUSIONS: Colonic involvement of Crohn's disease is common. Colonoscopy may be valuable in establishing the diagnosis and in assessing the extent and severity of colonic involvement, and CDEIS was value in the follow up of patients. Colonoscopic biopsies are helpful for verification of the diagnosis in difficult cases. Colonoscopy has replaced radiology as the initial test of choice in many clinical situations. PMID- 16313775 TI - [Dynamic development of normal fetal circulating blood hematopoietic stem progenitor cell surface antigen in on-going pregnancy]. AB - OBJECTIVE: To investigate the dynamic development of fetal circulating hematopoietic stem/progenitor cell (HSPC) surface antigen CD133, CD34 and CD38 in on-going normal pregnancy. METHODS: 0.2-2.0 ml fetal blood samples were obtained from 106 human fetuses between the gestational age of postmenstrual weeks 12 and 41, 96 being collected by ultrasound-guided percutaneous cordocentesis, 3 by ultrasound-guided selective feticide, and 7 being collected after birth. The nucleated cells (NCs) were separated, and the cell surface antigen CD34, CD38, and CD133 were labeled by bicolor immunofluorescence technique. Fluorescence activated cell sorting (FACS) was performed to analyze the concentration of CD34(+)/NC, CD38(-)/CD34(+), CD133(+)/NC, and CD133(+)/CD34(+) cell respectively. RESULTS: All operations were uneventful for the fetuses and mothers. The proportions of CD34(+)/NC, CD38(-)/CD34(+)cells, CD133+/NC, and CD133(+)/CD34(+) cells dramatically decreased when the gestational age advanced from week 12 to week 41. CD34+ cell/NC proportion decreased from 4.21% to 0.04% (r(2) = 0.90, x+/ s = 1.54% +/- 0.54%) with the peak at gestational week 12 and a the second peak at the week 20. (2) The percentage of CD38(-)/CD34(+) cells decreased from 58.5% to 10.7% (x+/- s = 28.66% +/- 9.88%) with advancing gestational age (r(2) = 0.82) with the peak at gestational week 12 and a second peak at week 22. (3) The percentage of CD133(+) cells among CD34(+) cells decreased with advancing gestational age, from 87.6% to 48.5% (x+/- s = 63.5% +/- 11.4%). (4) CD133(+) cells/NC concentration decreased with advancing gestational age, from 3.69% to 0.31% (x+/- s = 1.40% +/- 0.86%) with the peak time at gestational week 12., and showed a negative linear correlation with the gestational age (r(2) = 0.76). CONCLUSION: The immunophenotype of normal fetal circulating HSPC changes along with the gestational age in on-going pregnancy. The earlier the time, the more primitive the immunophenotype of the fetal circulating HSPCs. The more primitive fetal circulating HSPCs may be the ideal target for in utero gene therapy. PMID- 16313776 TI - [Effect of FK506 on axonal regeneration of rat sciatic nerve in regeneration chamber: an experimental study]. AB - OBJECTIVE: To evaluate the effect of FK506 on expediting nerve regeneration of rat Sciatic Nerve in regeneration chamber and to look for a proper way of its administration to treat peripheral nerve injuries. METHODS: Sixty adult male SD rats which were randamizely divided into 3 groups received a neurotomy to bilateral sciatic nerves, then we reconnected the broken nerves with silicon tube to make regeneration chambers. The regeneration chambers were filled with either normal saline (group A and group B) or 1 microg/ml FK506 (group C). The rats of group B also received daily injection of FK506 (1 mg/kg) at the back of the neck for 14 days. Local immunoreaction, weight of fresh gastrocnemius muscles, histological changes and electrophysiology were observed at designate time after neurotomy. RESULTS: At 6 weeks postoperation the extent of local lymphocytes infiltration in group B and C were less than that in group A, all results in group B were much better than that of group A. Results of group C were better than that of group A without significance. CONCLUSIONS: (1) Systemic administration of FK506 (1 mg/kg) showed neuroprotective and neurotrophic effect, which can facilitate nerve regeneration and promote functional recovery. (2) Local administration of FK506 (1 microg/ml) showed some extent of neuroprotective effect at early period of nerve injury, but the neurotrophic function is uncertain and still needs to be studied further. PMID- 16313777 TI - [Change of gene expression pattern and regulation of SFK on the change in early stage after hemitransection of the spinal cord in rat]. AB - OBJECTIVE: To investigate the changes of gene expression in early stage of hemitransection of spinal cord and the regulatory effects of the traditional Chinese mixture Suifukang (SFK) on these changes. METHODS: Eighty-six adult male SD rats were randomly divided into 3 groups: Suifukang group (Group S, n = 31, administered with SFK 3 days before the hemitransection at the dose of 2.5 g crude drugs/100 kg body weight QD and then 4 h and 8 h after hemitransection), model group (Group M, n = 45, administered with normal saline in the manner as in Group S), and normal control group (Group N, n = 10, without any treatment). The rats of Groups S and M underwent hemitransection of spinal cord at the level T12 three days before treatment. Twenty-four hours after the spinal cord injury (SCI), the rats were anesthetized and the injured tissues of Groups S and M and the corresponding tissues of Group N were removed. Total RNA was extracted, mRNA was purified and marked by Cy3 or Cy5 fluorescence labeling to synthesize cDNA hybridized probe that was hybridized with Biostars gene chip. Then the chip was scanned with ScanArray500 scanning apparatus. The data were analyzed by Imagene3.0 software. RESULTS: There were 139 differentially expressed genes in Group M, 83 up-regulated and 56 down-regulated, involved in multifarious functions, such as cell metabolism, especially lipid metabolism, and regulation of ionic channel, immunity and defense, cytoskeleton protein, extracellular matrix, cell cycle, etc. There were 37 differentially expressed gene in Group S, 11 up-regulated, such as such Anxal, Ca(2+)-ATPase, IL-1 beta, Ofp, and 26 down regulated, such as HSP-70 and Psap, several being key genes related to protection and regeneration of neuron. CONCLUSION: In the early stage after SCI many genes related to nervous degeneration, necrosis, traumatic inflammation, secondary injury of spinal cord, and aborted regeneration, etc, are expressed differentially. SFK regulates the changes of gene expression induced by SCI, thus initiating the protective effect of nervous tissues and promoting and supporting the regeneration of injured nervous tissues. PMID- 16313778 TI - [Effect of exogenous carbon monoxide on sequestration of polymorphonuclear neutrophils in the lung following limb ischemia-reperfusion: an experimental study]. AB - OBJECTIVE: To investigate the effect of exogenous carbon monoxide (CO) in inhibiting the sequestration of polymorphonuclear neutrophils (PMNs) in the lung following limb ischemia-reperfusion (IR) and the mechanism thereof. METHODS: PMNs of peripheral blood were isolated from the venous blood of a healthy volunteer. Serum was collected from a patient undergoing bilateral knee joint replacement as IR serum. Human pulmonary microvascular endothelial cells (PMVECs) were cultured and divided into 4 groups: control group (cultured under the condition of room air containing 5% CO2 for 5 h and cultured in normal human serum instead of medium during the last 4 hours of experiment), IR group (cultured under the condition of air containing 5% CO2 for 5h and cultured in the serum of IR patient during the last 4 hours), IR + CO group (cultured under the condition of air containing 0.025% CO and 5% CO2 for 5 hours and cultured in IR serum during the last 4 hours), and control + CO group (cultured under the condition of air containing and 0.025% CO and 5% CO2 for 5 hours and cultured in normal human serum during the last 4 hours). Immunofluorescence flow cytometry was used to detect the expression of intercellular adhesion molecule (ICAM)-1 and integrin CD11b in the PMVECs. Human PMVECs were put into the wells of a 96-well plate and added with PMNs to calculate the PMVEC-PMN adhesion rate. Tourniquettes were bound at the bilateral hind thighs of 32 healthy male SD rats for 4 hours so as to establish a rate IR model. The rats were randomly divided into 4 equal groups: control group (undergoing the same operation without causing limb ischemia and exposed to room air), IR group (undergoing bilateral hind limb ischemia for 4 h and reperfusion for 4 h and exposed to room air), IR + CO group (exposed to the containing 0.025% CO one hour before reperfusion till 4 hours after reperfusion), and control + CO group (exposed to air containing 0.025% CO at the corresponding time point as that of the IR + CO group). Then the rats were killed and their middle pulmonary lobes were taken out for microscopy and calculation of the number of PMNs in alveolar septum. Western blotting was used to examine the ICAM 1 protein expression in the lung. RESULTS: The ICAM-1 expression and integrin CD11b expression of the IR group PMVECs were significantly stronger than those of the IR + CO group PMVECs (both P < 0.05) and there were no significant differences in the ICAM-1 expression and CD11b expression between the control + CO and control groups (both P > 0.05). The PMN-PMVEC adhesion rate of the IR group PMVECs was 30 +/- 2.9%, significantly higher than those of the IR + CO group and control group PMVECs (19.8 +/- 1.5% and 13.4 +/- 1.1% respectively, both P < 0.05) and there was no significant difference in the PMN-PMVEC adhesion rate between the CO + control group and control group (P > 0.05). The lung tissues of the IR group rats showed edema and hemorrhage. The number of PMNs in the alveolar septum was 60.6 +/- 1.7/10 high power fields, significantly higher than those of the IR + CO group and control group (36.4 +/- 1.6 and 22.5 +/- 1.6 respectively, both P < 0.05) and there was no significant difference between the latter 2 groups (P > 0.05). The ICAM-1 protein expression in the lung of the IR group was the strongest, followed by the IR + CO group, control + CO group, and control group. CONCLUSION: Exogenous CO inhibits the limb/IR-induced PMN sequestration in the lung, probably by the mechanism of down-regulation of the expression of adhesion molecules and suppression of the PMN?PMVEC adhesion following IR. PMID- 16313779 TI - [Protection of oxymatrine against ischemia-reperfusion injury of liver and mechanism thereof: experiment with rats]. AB - OBJECTIVE: To investigate the effects of oxymatrine on protecting the liver against ischemia-reperfusion injury (IRI) and explore the mechanism thereof. METHODS: Thirty male Wistar rats were randomly divided into 3 equal groups: IRI group (2 ml normal saline was injected into the dorsal vein of penis, 30 min later laparotomy was performed, arterial clamp was used to grip the hepatic artery and portal vein for 30 minutes and then removed, the vessels were reperfused for 90 min, and 4 ml blood was collected from the aorta; parts of the liver were resected); oxymatrine group (oxymatrine 40 mg/kg was injected into the dorsal vein of penis, and the other procedures were the same as in the IRI group); and sham operation group (2 ml normal saline was injected into the dorsal vein of penis, laparotomy was performed, 150 min after the injection 4 ml blood was collected from the aorta and parts of the liver were resected). The levels of alanine transaminase (ALT) and aspartate transaminase (AST) were detected. The liver tissues underwent HE staining and TUNEL staining for pathological examination. Suspension of single hepatocytes was prepared to observe the ratio of apoptotic cells and cell cycles by flow cytometry (FCM). Western blotting was used to examine the Fas protein expression. RESULTS: The AST and ALT levels of the IRI group were 1326 U/L +/- 211 U/L and 768 U/L +/- 175 U/L respectively, significantly higher than those of the sham operation group (112 U/L +/- 53 U/L and 55 U/L +/- 17 U/L, both P < 0.05) and those of the oxymatrine group (513 U/L +/- 96 U/L and 352 U/L +/- 72 U/L respectively, both P < 0.01). The liver cells of the sham operation group were normal, those of the IRI group showed remarkable edema and cytoplasm degeneration. TUNEL staining showed remarkably more apoptotic cells in the IRI group. FCM showed that the apoptotic rate of hepatocytes was 42.8% +/- 5.2% in the IRI group, significantly higher than in the oxymatrine group (8.8% +/- 1.8%, P < 0.01), and that the ratio of hepatocytes in G(0)/G(1) stage of the IRI group was 99.2% +/- 1.8%, significantly higher than that of the sham operation group (77.0% +/- 2.1%), and that of the oxymatrine group (87.6% +/ 2.8%) (both P < 0.05); the ratio of hepatocytes in the S stage of the IRI group was 0.52% +/- 0.25%, significantly lower than those of the sham operation group (23.94% +/- 1.84%) and oxymatrine group (12.42% +/- 0.46%) (both P < 0.01). The Fas protein expression was significantly highly in the IRI group than in the oxymatrine group. CONCLUSION: Remarkably reducing the IRI of hepatocytes, oxymatrine has potential to protect the liver against IRI during surgical intervention. PMID- 16313780 TI - [Effects of interleukin-18 on asthmatic airway inflammation: experimental study of guinea pig asthmatic model]. AB - OBJECTIVE: To investigate the effects of Interleukin-18 (IL-18) on asthmatic airway inflammation. METHODS: Thirty healthy adult male guinea pigs were randomly divided into 3 equal groups: asthmatic model group (Group A, undergoing intraperitoneally injection of ovalbumin (OVA) once and spraying of OVA aerosol once a day for 5 days; control group (Group B), undergoing intraperitoneally injection of OVA once and spraying of normal saline aerosol once a day for 5 days; and interleukin (IL)-18 intervention group (Group C, undergoing intraperitoneally injection of OVA once and intraperitoneal injection of IL-18 on the days 1, 3, 8. 10. 15, 17, and 19. Twenty-four hours after the final spraying or IL-18 injection the bronchalveolar lavage fluid (BALF) of the left lungs were obtained. HE staining was conducted to the sediment to examine the numbers of eosinophils, neutrophils, and monocytes. ELISA was used to detect the Th1/Th2 cytokines in the BALF. The left lungs underwent pathological examination. RESULTS: The number of EOS in BALF of Groups A was (98 +/- 58) x 10(6)/L, significantly higher than those of Group B, (12 +/- 10) x 10(6)/L, and Group C, (29 +/- 10) x 10(6)/L (P < 0.01 and P < 0.05). The numbers of neutrophils in the BALF of Group A was (24 +/- 16) x 10(6)/L, significantly higher than those of Group B and C [(9 +/- 7) x 10(6)/L and (10 +/- 5) x 10(6)/L respectively, both P < 0.05]. The concentration of IFN-gamma and IL-2 in group A were both significantly lower than those of Group B and Group C (P < 0.05 and P < 0.01). The concentration of IL-4 in Group A was significantly higher than those of Groups B and C (both P < 0.05). The concentration of IL-5 of Group A was significantly higher than those of Group Bs and C (both P < 0.01). CONCLUSION: IL 18 effectively inhibits asthmatic airway inflammation by regulating the Th1/Th2 balance. PMID- 16313790 TI - [The level of surgical treatment for lung cancer in China]. PMID- 16313791 TI - [Theory of jaundice treatment for liver transplantation]. PMID- 16313792 TI - [Therapeutic effects of recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand on non-small lung cell cancer: an experimental with rats]. AB - OBJECTIVE: To evaluate the therapeutic effects of recombinant mutant human tumor necrosis factor-related apoptosis-inducing ligand (rmhTRAIL) on non-small cell cancer (NSCLC). METHODS: NSCLC cells of the line NCI-H460 were cultured and underwent agarose gel electrophoresis. rmhTRAIL of different concentrations was added into the culture fluid to observe the influence of rmhTRAIL. Nude BALB/c rats were transplanted with rat NCI-H460 tumor. Then the rats carrying xenografts were randomly divided into 5 groups of 8 rats: negative control group to be injected intraperitoneally with normal saline; positive control group to be injected with vancristine; low-dose rmhTRAIL group, to be injected with 1.7 mg/kg rmhTRAIL, middle-dose rmhTRAIL group to be injected with rmhTRAIL 5.0 mg/kg, and high-dose rmhTRAIL group to be injected with 15.0 mg/kg rmhTRAIL. The size of tumor was measured every 3 approximately 4 days. Ten days after the administration of different drugs the rats were killed and the tumors were taken out to undergo TUNEL staining for microscopy. RESULTS: The relative tumor volume of the low-dose rmhTRAIL group and high-dose rmhTRAIL group were 3.19 +/- 2.05 and 1.47 +/- 0.77 respectively, both significantly smaller than that of the negative group (8.48 +/- 5.87, P < 0.05 and P < 0.01). The relative tumor growth rate of the low-dose rmhTRAIL group and high-dose rmhTRAIL group were 37.6% and 17.3% respectively. The tumor weight of low-dose rmhTRAIL group and high-dose rmhTRAIL group were 1.09 g +/- 0.55 g and 0.31 g +/- 0.09 g respectively, both significantly lighter than that of the negative control group (2.78 +/- 0.77, both P < 0.01). Large amount of apoptotic cells were seen in the tumor tissues of the rmhTRAIL -treated rats and tumors cells cultured with rmhTRAIL. Agarose gel electrophoresis showed apoptosis-characteristic ladder in the DNA extracted from the rmhTRAIL -treated tumors cells. CONCLUSION: rmhTRAIL dose-dependently inhibit the growth of NSCLC cells, primarily by inducing apoptosis of tumor cells. PMID- 16313793 TI - [Clinical value of computed tomography and fluorine-18 fluorodeoxyglucose positron remission tomography in diagnosis of mediastinal metastasis of non small cell lung cancer]. AB - OBJECTIVE: To compare the clinical values of computed tomography (CT) and fluorine-18 fluorodeoxyglucose positron remission tomography (FDG-PET) in diagnosis of mediastinal metastasis of non-small cell lung cancer (NSCLC). METHODS: Sixty patients with respectable NSCLC underwent CT and FDG-PET with an interval of 2 weeks and then underwent thoracotomy for clearance of the lymph nodes or biopsy of the mediastinal lymph nodes via mediastinoscopy. The specimens of mediastinal lymph node underwent HE staining and PCNA/Ki67 immunohistochemical staining. The sensitivity, specificity, accuracy, positive prediction value, and negative prediction value in diagnosis of metastasis of mediastinal lymph nodes of these 2 procedures were compared. RESULTS: The sensitivity, specificity, accuracy, positive prediction value, and negative prediction value in diagnosis of metastasis of mediastinal lymph nodes were 92.3%, 87.5%, 91.2%, 96.0%, and 77.7% respectively for FDG-PET, and were 76.9%, 50.0%, 70.6%, 83.3%, and 40.0% respectively for CT. The sensitivity rate was 98% for CT plus FDG-PET. CONCLUSION: PET is superior to CT in diagnosis of metastasis of NSCLC to mediastinal lymph nodes. PET + CT significantly increases the sensitivity in diagnosis. PMID- 16313794 TI - [Combined therapy for stage IIIa non-small cell lung cancer]. AB - OBJECTIVE: To evaluate the long-term outcome of combined therapy for stage IIIa non-small cell lung cancer (NSCLC). METHODS: Eight and twelve patients with stage IIIa NSCLC, 662 males and 150 females, aged 58.6 (24-79), underwent surgical resection, 326 of which received pre- and postoperative chemotherapy and/or radiotherapy, and 486 of which received surgery alone. The patients were classified according to the TNM staging, and the specimens of resected cancer were examined pathologically. The clinical data of these patients were analyzed. RESULTS: The overall 5-year survival rate was 16.4%. The 5-year survival rate of the combined therapy group was 23.5%, significantly higher than that of the surgical resection alone group (P < 0.01). The 5-year survival rate was 27.6% for the squamous cell carcinoma (SCC), and 23.5% for adenocarcinoma (AC) in the combined therapy group; and was 115.2% for SCC and 9.9% for AC in the surgical resection alone group (all P < 0.01). CONCLUSION: The curative effectiveness of combined therapy is significantly better than surgical resection alone for stage IIIa NSCLC. However, The combination should be based on the TNM staging and pathological type. PMID- 16313795 TI - [Duffy antigen receptor for chemokines attenuates breast cancer growth and metastasis: an experiment with nude mice]. AB - OBJECTIVE: To study the effects of Duffy antigen receptor for chemokines (DARC) on the tumorigenesis and metastasis of breast cancer. METHODS: Human breast cancer cells of the line MDA-MB-435HM with a great potentiality of metastasis were cultured and then divided into 3 groups: MDA-MB-435HM-DARC cell group, to be transfected with pcDNA3/DARC FyB containing human DARC cDNA; MDA-MB-435HM-vect cell group, to be transfected with blank plasmid pcDNA3; and MDA-MB-435HM cell group without transfection. Female BALB/c nude mice were implanted with MDA-MB 435HM cells into the nipple. The size of the implanted cancer was measured once a week. The mice were killed 4, 6, and 8 weeks after the implantation respectively and their lungs were taken out to observe the number of metastatic tumor. Immunohistochemical staining was performed to calculate the microvascular density (MVD). Western blotting was used to detect the matrix metalloproteinase (MMP)-9 protein expression of the lung tissues. ELISA was used to detect the concentrations of the 2 DARC ligands: CNCL8 and CCL2 in the supernatant of culture fluid of the 3 kinds of cells and the metastatic tumors. RESULTS: The expression levels of DARC mRNA and protein of the MDA-MB-435HM cells were 38% those of the MDA-MB-435 cells. The expression levels of CXCL8 and CCL2 of the MDA MB-435HM-DARC cells were significantly lower than those of the MDA-MB-435HM-vect cells and MDA-MB-435HM cells (both P < 0.05). All mice developed tumor with a tumorigenesis rate of 100%. However, the tumor occurred 17 days after implantation in those mice implanted with MDA-MB-435HM-DARC cells, significantly later then in those implanted with MDA-MB-435HM-vect cells and MDA-MB-435HM cells (9 and 10 days after implantation) the size of tumor in the former group being significantly smaller than those of the 2 latter groups (both P < 0.01). The number of metastatic foci in the lung of the MDA-MB-435HM-DARC group was significantly less than those of the other 2 groups (both P < 0.01). The level of CCI2 of the implanted tumor in the MDA-MB-435HM-DARC group was significantly lower than those of the other 2 groups (both P < 0.01). The MVD of the MDA-MB 435HM-DARC was significantly less than those of the other 2 groups (both P < 0.01) and most of the vessels of the MDA-MB-435HM-DARC group were not newly formed vessels with a diameter < 8 red blood cells. The MMP-9 protein expression levels of the MDA-MB-435HM-vect cells and MDA-MB-435HM-cells were 3.1 and 3.4 times that of the MDA-MB-435HM-DARC group (both P < 0.05). CONCLUSION: DARC, a chemokine decoy receptor, inhibits the chemokine-induced angiogenesis, thus inhibiting the growth and lung metastasis of breast cancer. PMID- 16313796 TI - [Developing a hospital-based high risk scoring model and screening strategy for pancreatic cancer]. AB - OBJECTIVE: To develop a high risk scoring model and screening strategy to improve the diagnosis of symptomatic pancreatic cancer. METHODS: A hospital-based case control study was undertaken among a cohort comprising 136 pancreatic cancer patients and 191 patients with benign gastrointestinal diseases who were hospitalized between Feb, 2002 and Mar, 2004. All patients were consulted with an epidemiological questionnaire. Risk factors and symptoms described in the questionnaire were compared between these two groups. Significant and borderline risk factors and symptoms were selected to undergo multivariate logistic regression. A high risk scoring model was constructed according to the weighted numerical scores of every variable. The diagnostic values of 4 tumor markers of pancreatic cancers (serum CA19-9, CA242, stool K-ras and p53 mutation) and 2 imaging tests (abdominal spiral CT and ultrasonography) were evaluated to provide evidence for establishing the diagnostic strategy. RESULTS: The average score was significantly higher for the pancreatic cancer patients than for the control patients [mean 49.6 (95% CI: 45.6-53.7) vs 21.6 (95% CI: 19.3-23.9); P < 0.01]. With a cutoff value of 27 points, the sensitivity and specificity of the scoring model was 87.0% and 68.1% respectively. CT had the highest sensitivity (94.7%) among the 4 tumor markers and 2 imaging tests. Combination of the two tumor markers (CA19-9 and stool K-ras) with CT or ultrasonography could improve the sensitivity to 100% with a specificity of 67.5%-73.0%. It was suggested that for high risk patients with a risk score more than 27, the combination test be recommended as the primary test, endoscopic ultrasonography (EUS) and/or endoscopic retrograde cholangipancreatography (ERCP) be considered for patients with inconclusive CT studies when risk score and tumor markers nevertheless suggest pancreatic cancer. CONCLUSION: The high risk scoring model provides a simple and feasible way to screen pancreatic patients in hospitals at all levels. Once high risk patients are identified, they can be transferred to higher level hospitals to receive further examinations. This screening strategy may help detect more resectable pancreatic cancers. PMID- 16313797 TI - [In-hospital effect of primary percutaneous coronary intervention on aged patients with acute myocardial infarction and diabetes mellitus]. AB - OBJECTIVE: To evaluate the in-hospital effect of emergency percutaneous coronary intervention (PCI) on aged patients with acute myocardial infarction (AMI) and diabetes mellitus. METHODS: The clinical data of 556 patients aged > or = 60 with AMI hospitalized from December 1995 to April 2005, who underwent emergency PCI within 24 hours after the onset, including 127 cases complicated with diabetes mellitus (Group DM) and 429 cases without diabetes (non-diabetes mellitus group, Group ND), were analyzed respectively. RESULTS: There were not significant differences in age, sex, smoking rate, hypertension rate, myocardial infarction (MI) rate, and time between onset and treatment between these 2 groups. However, the percentages of anterior wall AMI rate, non-ST elevation myocardial infarction, Killip III-IV grade, and intra-aortic balloon pump (IABP) application of Group DM were 58.3%, 13.4%, 27.6%, and 16.5%, all significantly higher than those of Group ND (46.4%, 6.8%, 27.6%, and 8.6%, P < 0.001, 0.02, 0.03, and 0.01). In Group DM, 59.1% of the infarction related artery was left anterior descending branch, and in Group ND 45.0% of the infarction related artery was left anterior descending branch (P < 0.05). The rate of triple vessel disease in Group DM was 77.2%, significantly higher than that of the Group ND (60.8%, P < 0.01). There were no significant differences in the PCI immediate success rate, intra-operational no-reflow rate, post-PCI complication rate and mortality during hospitalization between the two groups. CONCLUSION: Emergency in-hospital PCI is effective on aged AMI patients with diabetes mellitus. PMID- 16313798 TI - [Effects of renal artery stenting on renal function of patients with ischaemic nephropathy]. AB - OBJECTIVES: To evaluate the effects of stent revascularization on renal function of patients with atherosclerotic renal artery stenosis with renal insufficiency. METHODS: Percutaneous transluminal renal angioplasty with stent (PTRAS) was performed on 27 consecutive patients with severe atherosclerotic renal artery stenosis for preservation of renal function, and follow-up was conducted for 6 approximately 48 months. RESULTS: Technically PTRAS was successfully performed in all 27 patients. The creatinine value was 155 micromol/L +/- 31 micromol/L before operation, and increased to 189 micromol/L +/- 38 micromol/L (P < 0.001), then decreased to 145 micrommol/L +/- 22 micrommol/L, 143 micromol/L +/- 22 micromol/L, 143 micromol/L +/- 24 micromol/L, and 139 micromol/L +/- 10 micromol/L, 6, 12, 24, and 36 months after the PTRAS respectively (all P < 0.05). The severe complications related to the procedure mainly included 7 cases of acute renal failure, 4 cases being reversible. One case died of cardiac infarction with underlying coronary heart disease 9 months after and 1 case died heart failure due to valvular disease of heart 25 months after. CONCLUSION: Renal artery stent revascularization is effective for preservation of renal function in patients with ischaemic nephropathy effect on. However, complications related to the procedure frequently occurred. The long-term efficacy is to be investigated further. PMID- 16313800 TI - [Swanson double-stem silicone implant arthroplasty of first metatarsophalangeal point]. AB - OBJECTIVE: To evaluate the curative effectiveness of Swanson double-stem silicone implant arthroplasty on the reconstruction of first metatarsophalangeal joint. METHODS: Swanson double-stem silicone implant arthroplasty was performed on 10 patients, 2 males and 8 females, aged 60 (57-75), suffering from diseases of first metatarsophalangeal joint with 13 diseased joints, including hallux valgus with geriatric osteoarthritis (8 cases, 10 joints), rheumatoid arthritis (1 case, 2 joints), and traumatic arthritis (1 case, 1 joint). All patients were followed up for an average of 25 months (12-38 months). RESULTS: Eight patients felt complete satisfaction with the operation, 1 patient showed satisfaction to some extent, and 1 patient was not satisfied because of pain of the first matatarsophalangeal joint due to severe hyperosteogeny surrounding the cut bone face 3 years after the operation. Osteolysis around the silicone implant occurred in 2 cases without clinical symptoms, and no special treatment was given. CONCLUSION: With the advantages of alleviating pain, preserving the length and alignment of great toe, improving the function of walking, and correcting the deformity, Swanson double-stem silicone implant arthroplasty is effective on reconstruction of the first metatarsophalangeal joint. PMID- 16313801 TI - [Isolation and identification of chronic myelogenous leukemia bone marrow mesenchymal stem cells and their functional characteristics]. AB - OBJECTIVE: To isolate and culture bone marrow mesenchymal stem cells (MSCs) from chronic myelogenous leukemia (CML) patients and examine their functional characteristics. METHODS: Bone marrow was extracted from the anterior superior iliac spines of 21 patients with CML. MSCs were isolated and cultured. Single colony derived MSCs were harvested by limiting dilution. The cell cycle and immunophenotype of the expanded clonal MSCs were detected by fluorescence activated cell sorter (FACS). Different agents were used to induce the MSCs to differentiate into osteocyte, adipocyte and neural cells. Von Kossa staining, oil red staining, and Western blotting was used to examine the ability of differentiation. PCR was used to detect the expression of BCR/ABL gene. The ultrastructure of the CML derived MSCs was observed with electron microscopy. Sixteen BALB/c nude mice were randomly divided into 2 equal groups to be inoculated with HL60 cancer cells and CML derived MSCs to observe the tumorigenicity. MSCs were cultured in soft agar for 2 weeks to observe the clone growth. RESULTS: Fibroblast-like, positive in CD29, CD44, and CD105, and negative in CD116, CD34, CD48, and HLA-DR, the CML derived MSCs could differentiate into osteocyte, adipocyte and neural cells. CML derived MSCs showed normal karyotype and ultrastructure, they did not express BCR/ABL gene. After 2 weeks' culture no clone was formed from the MSCs. Four weeks after tumors were shown in 6 of the 8 mice inoculated with HL60 cells, and no tumor was seen in the mice inoculated with MSCs. CONCLUSION: Able to differentiate into different types of cell and without tumorigenicity, MSCs from the bone marrow of CML patients have the potentiality in clinical application. PMID- 16313802 TI - [Proteasomal inhibitor lactacystin induces cell apoptosis and caspase 3 activation in PC12 cells]. AB - OBJECTIVE: To explore whether proteasome dysfunction cam induce dopaminergic cell apoptosis and investigate the probable molecular mechanism. METHODS: MTT assay was applied to measure the cell vitality of rat pheochrom-ocytoma cells of the Line PC12 exposed to highly specific proteasomal inhibitor lactacystin (0, 1 micromol/L, 5 micromol/L or 10 micromol/L) for 24 hours. After the PC12 cells were treated with 10 micromol/L lactacystin for 24 hours, apoptosis was estimated by Hoechst fluorescence staining and flow cytometry. When the PC12 cells were treated with 10 micromol/L lactacystin for 0, 24 or 48 hours, the level of caspase 3 cleaved fragments were analyzed by Western blotting. RESULTS: The PC12 cells exposed to 5 micromol/L or 10 micromol/L lactacystin for 24 hours showed a significant decrease in cell vitality (P < 0.05). Following treated with 10 micromol/L lactacystin for 24 hours, PC12 cells were seen to be nuclear condensation and fragmentation consistent with an apoptotic nuclear morphology by were seen in the Hoechst fluorescence staining and confirmed to have a significant increase of apoptotic cells (about 31.4%) by flow cytometry. Western blotting showed that there was a very low level of caspase 3 cleaved fragments (17,000) in control cells. But, after PC12 cells were exposed to 10 micromol/L lactacystin for 48 hours, the protein level of caspase 3 cleaved fragments (17,000) increased obviously. CONCLUSION: Proteasomal inhibitor lactacystin leads to dopaminergic cell apoptosis. The activation of caspase 3 protease may contribute to the mechanism of lactacystin-induced apoptosis in PC12 cells. Proteasome dysfunction may play an important role in the pathogenesis of Parkinson's disease. PMID- 16313803 TI - [Static pressure induces vascular smooth muscle cells proliferation and apoptosis]. AB - OBJECTIVE: To study the role of static pressure on proliferation and apoptosis of the vascular smooth muscle cells. METHODS: Vascular smooth muscle cell line A10 were cultured at normal atmosphere, 80 mmHg, 100 mmHg, 200 mmHg static pressure respectively. Cell viability was determined by MTT assay, cell cycle and apoptosis rate were analyzed by flow cytometer. RESULTS: After exposure to lower static pressure (100 mmHg) for 48 hour, cell viability and proliferation index (PI) of A10 cell significantly increased, the percentage of A10 cell in G(0)/G(1) phase decreased significantly and the apoptosis rate showed no difference as compared with those exposed to atmospheric pressures. However after exposure of A10 cell to higher static pressure (200 mmHg) for 48 hours, cell viability and proliferation index (PI) significantly decreased, the percentage of A10 cell in G(0)/G(1) phase increased significantly, apoptosis rate significantly increased, as compared with cells exposed to atmospheric pressure. CONCLUSION: Lower static pressure can facilitate vascular smooth muscle cells proliferation, while higher pressure can induce cell apoptosis. PMID- 16313804 TI - [Investigating the inhibitory effects of interferon-alpha on the replication of hepatitis C virus replicon]. AB - OBJECTIVE: To investigate the inhibitory effects of interferon-alpha (IFN-alpha) on HCV replicon and evaluate the expressing levels of signal transducer and activator of transcription genes (STAT1 and STAT2) and IFN-alpha stimulated genes (ISGs) which may mediate the inhibitory effects of IFN-alpha on HCV. METHODS: Firstly HCV replicon cell culture system was established by transfecting HCV replicon RNA transcribed in vitro into Huh7 cells and screening with G418. Secondly, the established HCV replicon cells were treated with various concentrations of IFN-alpha (0, 10, 25, 50, 100, 250, 500, 750, 1000, 2500, and 5000 IU/ml) for 72 h or treated with 1000 IU/ml of IFN-alpha for different lengths of time (0, 24, 48, 72, 96 h), then the levels of HCV RNA and NS5A protein in these cells were examined by semi-quantitative RT-PCR and Western blot respectively. RESULTS: IFN-alpha could effectively inhibit HCV RNA replication. 10 IU/ml or 25 IU/ml of IFN-alpha could lead to about 68% and 75% of HCV-RNA reduction respectively. The cells treated with 1000 IU/ml IFN-alpha for 24 h or 96 h had about 75% and 88% of HCV RNA reduction compared with the cells of control, demonstrated that the inhibitory effects of IFN-alpha on HCV replicon are in dose and time dependent manners. The expressions of antiviral ISGs-PKR, 2'5'OAS, G1P3, ISG20 and ISGF3gamma are strongly induced by IFN-alpha. CONCLUSIONS: HCV replicons were sensitive to IFN-alpha treatment. The inhibitory effects of IFN-alpha on HCV RNA and NS5A are both dose and time dependents. PKR, 2'5'OAS, G1P3, ISG20 and ISGF3gamma may mediate the inhibitory effects of IFN alpha on HCV replicon replication. PMID- 16313806 TI - [The effect of tubular epithelial cells activated by aldosterone on renal interstitial fibroblasts in co-culture system]. AB - OBJECTIVE: Local aldosterone (ALDO) could be synthesized by human proximal tubular epithelial cell lines (HKC) after the stimulation of endothelin-1 (ET-1) in vitro. T to observe the effect of tubular epithelial cells activated by aldosterone (ALDO) on renal interstitial fibroblasts in co-culture system. METHODS: (1) Human Proximal tubular epithelial cells of the line HKC were stimulated with ALDO at different concentrations and times, then reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) were performed to detect TGF-beta1 expression; (2) HKC were co stimulated with ET-1 (10(-9) mol/L) and spironolactone at different concentrations and times to evaluate the influence of endogenous ALDO on TGF beta1 expression; (3) HKC which were activated by 10(-7) mol/L ALDO for 12 h, and hRIFs were co-cultured for 48 h with or without anti-TGF-beta1 antibody (1.0, 2.0 microg/ml) in the media, then the production of type I collagen (Col-I) in the cell layer of hRIFs was detected by ELISA. RESULTS: (1) After stimulation with ALDO, the expression of TGF-beta1 by HKC was up-regulated in a dose-and time dependent manner. With 10(-9) or 10(-7) mol/L ALDO stimulation (mRNA determination at 12 h and protein at 48 h), the expression of TGF-beta1 was significantly increased (vs 0 mol/L ALDO, P < 0.05 or 0.01). With 10(-7) mol/L ALDO stimulated at different times (mRNA determination at 8 h, 12 h and 16 h and protein at 12 h, 24 h and 48 h), the expression of TGF-beta1 was also significantly increased (vs 0 h, P < 0.05 or 0.01). (2) After co-stimulation with ET-1 and spironolatone, the expression of TGF-beta1 by HKC was down-regulated in a dose-and-time dependent manner along with spironolatone. The expression of TGF beta1 mRNA and protein was decreased in the 10(-9) or 10(-7) mol/L spironolatone groups compared with 0 mol/L group (P < 0.05 or 0.01). The expression of TGF beta1 (mRNA and protein) was significantly decreased in 10(-9) mol/L ET-1 and 10( 7) mol/L spironolatone co-stimulation group compared with 10(-9) mol/L ET-1 stimulated group. (3) The production of Col-I by hRIFs which were co-cultured with activated HKC by ALDO, was significantly increased (vs control group in which hRIFs co-cultured with normal HKC, P < 0.01), and this effect was partially inhibited by 1.0 or 2.0 microg/ml anti-TGF-beta1 antibody (P < 0.05). CONCLUSION: The expression of TGF-beta1 in HKC cells can not only be up-regulated by exogenous ALDO, but also can be up-regulated by endogenous ALDO in an autocrine manner. The HKC cells activated by ALDO can promote the synthesis of Col-I in hRIFs by a "cross talking" way, and this effect is partially mediated by TGF beta1. PMID- 16313808 TI - [The research on the relationship between the polymorphism of 1082A/G, anti inflammatory interleukin-10 gene promoter with its effect of preventing ESRD patients from microinflammation and arteriosclerosis]. AB - OBJECTIVE: We do this investigation in order to reveal the relationship between the polymorphism of 1082A/G, anti-inflammatory interleukin-10 gene promoter, and end stage renal disease (ESRD) patients microinflammatory state and arteriosclerosis (AS). METHODS: We used PCR-RFLP to measure the various kinds of distribution of IL-10 gene-1082A/G genotype and relevant indexes of microinflammatory state and AS of 870 ESRD patients and 1000 healthy persons of control group and to analyze the mechanism of its protection effect keeping ESRD patients away from microinflammation and arteriosclerosis. RESULTS: Compared with the control group, CRP, TNF-alpha, CH50, C3, IL-10 and Alb of ESRD group were in the normal range, but still significantly higher than those of the control group, while IL-10, Alb were significant lower (P < 0.05). The genotype distribution and allele frequency of IL-10A/G gene had no significant differences between the healthy group and the control group (P > 0.05). Levels of CRP, TNF-alpha, CH50 and C3 of ESRD patients with IL-10A/A genotype were significantly higher than those of ESRD patients with G/G and G/A genotype (P < 0.05), while IL-10 and Alb were significantly lower (P < 0.01). The production of IL-10 in serum from patients with IL-10A/A genotype was significantly lower than that of patients with G/G and G/A genotype (P < 0.01). The incidence rate of AS of patients with IL-10-1082A/A genotype was significantly higher than that of patients with G/G and G/A genotype (P < 0.01). The raise of AS incidence rate was correspondent with the decline of serum IL-10 and raise of serum CRP and Fib. CONCLUSION: The IL-10A/A genotype is a predictable factor of microinflammatory state and high AS incidence rate in ESRD patients. We use IL-10G/G genotype to modulate the high production of serum IL-10, to decline inflammatory reaction and to keep away from microinflammation and AS in ESRD patients. We should work hard on improving the dialysis membrane to reduce the anti-inflammatory factors in uremia for chronic renal failure patients with high arteriosclerosis risk. PMID- 16313812 TI - [The rule and strategy of chemotherapy in gynecologic malignancy]. PMID- 16313813 TI - [New strategies for the management of advanced gastric cancer]. PMID- 16313815 TI - [Treatment of metastatics colorectal cancer to the liver]. PMID- 16313816 TI - [Strategy on drug tolerance of ovarian cancer therapy]. PMID- 16313817 TI - [A multicenter randomized clinical trial of weekly paclitaxel and carboplatin vs three weeks paclitaxel and carboplatin as first line chemotherapy in epithelial ovarian cancer]. AB - OBJECTIVE: To compare the efficacy of combination regiments of taxol given weekly plus carboplatin and taxol given every three weeks plus carboplatin. To observe the toxicity of the two regiments. To observe the two-year survival rate in the two groups. METHODS: Total 125 eligible patients in 13 centers of CGOG were entered into the two arms of this randomized clinical trial, of whom 51 were entered into weekly taxol group and 74 entered into 3 weeks taxol group. RESULTS: 81.6% (102/125) of patients had satisfactory decreasing of CA125 level after optimal cytoreductive surgery and chemotherapy. 86.3% (44/51) of patients is in weekly group and 78.4% (58/74) of patients in three weeks group (P > 0.05). Relapse frequency is 29.7% in every three weeks group and 19.6% in weekly group (P > 0.05). Median interval to relapse is 15.7 months in every three weeks group and 13.6 months in weekly group (P > 0.05). One-year survival rate is 95.2% in every three weeks and 93.9% in weekly group (P > 0.05). Two-year survival rate is 78.7% in every three weeks and 85.3% in weekly group (P > 0.05). Grade III and IV myelosuooression is 45.9% in three weeks group and, 27.5% in weekly group (P < 0.05). CONCLUSION: (1) The two regiments had equal efficacy. (2) Myelosuppression was less frequency in the weekly group than in every three weeks group. (3) Weekly taxol therapy has mild toxicity and is more suitable for the old and feeble patients. Weekly taxol therapy can be conveniently administered in outpatients department. PMID- 16313818 TI - [Clinical effects of combination chemotherapy with irinotecan hydrochloride and cisplatin on cervical cancer: study of 46 patients]. AB - OBJECTIVE: To evaluate the efficacy and toxicity of the combination chemotherapy of irinotecan hydrochloride plus cisplatin on cervical cancer. METHODS: Forty-six patients with cervical cancer, 43 with squamous cell carcinoma, 2 adenocarcinoma, and 1 small cell carcinoma, 41 being treated initially and 5 being recurrent cases of which 2 had undergone radiotherapy plus chemotherapy, 2 radical hysterectomy plus pelvic radiotherapy after operation, and 1 radical hysterectomy plus chemotherapy, aged 38 (22-61), were treated with irinotecan hydrochloride, 60 mg/m(2) in 250 ml normal saline administered on days 1, 8, and 15 by intravenous infusion over 60 min, plus cisplatin, 60 mg/m(2) in 500 ml 5% glucose given on day 1. This regimen was repeated every 28 days. Effectiveness evaluation was conducted after 1-2 courses for the initially treated patients and after 2-6 courses for the recurrent patients. RESULTS: Totally 79 courses of treatment were given to the 41 patients. After 1-2 courses 4 (9.8%) of the 41 stage Ib2-IIIb initially treated patients achieved complete remission, 30 (73.2%) achieved partial remission, and 7 (17.1%) remained at stationary phase with a overall effective rate of 82.9%. Of the 29 IIb and IIIb stage advanced patients who failed to receive operation originally 20 patients (69%) succeeded to be treated by radical hysterectomy after 1 to 2 courses when the tumorless space between the uterine and the pelvic wall > or = 3 cm with an operatibility rate of 69%. Of the 5 recurrent patients 1 case achieved complete remission, 2 partial remission, and 2 remained stable; none progression of disease was observed after 4-6 courses. The main toxic response and side effect included myelosuppression and diarrhea. CONCLUSION: Effective neoadjuvant chemotherapy on cervical cancer, combination therapy of irinotecan hydrochloride and cisplatin win opportunity for treatment and improves their life quality with tolerable side effects. PMID- 16313819 TI - [Effects of primary chemotherapy with single methotrexate on low-risk gestational trophoblastic neoplasia and influencing factors thereof]. AB - OBJECTIVE: To evaluate the effects of primary chemotherapy with single-agent methotrexate (MTX) on low-risk gestational trophoblastic neoplasia (GTN) and the influencing factors thereof. METHODS: Sixty-one GTN patients with the score of < or = 6 according to the new International Federation of Gynecology and Obstetrics (FIGO) scoring system (2000) were divided into 2 groups: 51 patients were treated with single MTX 0.4 mg/kg daily for 5 days (MTX 5 d group), and 10 patients were treated with MTX on the days 1, 3, 5, and 7, and with folinic acid (FA) 0.1 mg/kg on the days 2, 4, 6, and 8 (MTX + FA group), both group with an interval of treatment course of 2 weeks. The serum level of human chorionic gonadotropin (hCG) was detected every week. If a plateau or increase of serum hCG appeared between 2 examination results, meaning tolerance to MTX, the patients concerned had to undergo different regimens of salvage chemotherapy, all with MTX as one of their components. Univariate and multivariate methods were used to analyze the relationships of different factors to the outcomes of chemotherapy. RESULTS: Thirty-five of the 51 patients of the MTX 5d group (68.6%) achieved complete primary remission, 3 of the 10 patients of the MTX + FA group achieved complete primary remission, and all patients achieved complete remission after salvage chemotherapy. Univariate analysis showed that the mean pretreatment serum level of hCG, duration between antecedent pregnancy and start of treatment, size of tumor, FIGO score, specific regimen of MTX were significantly associated with outcome of chemotherapy (P = 0.004, 0.022, 0.017, 0.005, 0.021 respectively). Logistic regression analysis showed that only three independent factors predictive for the outcome of chemotherapy: MTX regimen (OR = 2.476), FIGO score (OR = 1.431), and pretreatment hCG titer (OR = 1.001). CONCLUSION: Primary chemotherapy with single MTX regimen may still be one of the options for patients with low-risk GTN according to the new FIGO scoring system, though the rate of complete primary remission appears to be lower. All patients with low-risk GTN achieve complete remission after salvage chemotherapy. MTX regimen, FIGO score, and pretreatment hCG are independent risk factors of outcome of single MTX chemotherapy. PMID- 16313820 TI - [Relationship between metastatic rate and amount of lymph nodes and prognosis of gastric cancer]. AB - OBJECTIVE: to investigate the relationship between metastatic rate and amount of lymph nodes and prognosis of gastric cancer. METHODS: The clinical data of 672 patients of gastric cancer with 22.7 regional metastatic lymph nodes on average were analyzed retrospectively. The patients were divided into 3 groups according to the TNM staging system (1997, 5th edition): pN0 (without metastatic lymph node), pN1 group (with 1-6 metastatic lymph nodes), pN2 group (with 7-15 metastatic lymph nodes), and pN3 group (with more than 215 metastatic lymph nodes), or into 4 groups according to the lymph node metastasis rate: rN0 group (without metastasis of lymph node), rN1 group (with a lymph node metastasis rate 0 < 10%), rN2 group (with a lymph node metastasis rate of 10%-30%), and rN3 group with a lymph node metastasis rate > 30%). The 5-year survival rates of different groups in these 2 classification systems and the relationship between metastatic rate and amount of lymph nodes and prognosis of gastric cancer were analyzed. RESULTS: According to the lymph node metastasis grading system there were 93 patients in the pN0 group, 201 in the pN1 group, 108 in the pN2 group, and 32 in the pN3 group with the 5-year survival rates of 90.3%, 65.2%, 37.0%, and 18.9% respectively (chi(2) = 12.56, P < 0.01). According to the lymph node metastasis system 93 patients were in the rN0 group, 183 in the rN1 group, 92 in the rN2 group, and 66 in the rN3 group, with the 5-year survival rates of 90.3%, 69.9%, 45.7%, and 10.6% respectively (chi(2) = 14.38, P < 0.01). If the cases in the same pN group were re-divided according to the lymph node metastasis rate, there were significant differences in the 5-year survival rate among the new rN groups (chi(2) = 5.26, P = 0.21), and there was a great difference in the distribution of prognosis among different groups. However, if the cases in the same group rN group were re-divided according to the amounts of metastatic lymph nodes, there was no significant difference in 5-year survival rate among different new pN groups (chi(2) = 0.14, P = 0.932). Multivariate COX model analysis showed that the influencing factors of the prognosis were rN grading, pN grading, infiltration depth of cancer, gross type, and size of the cancer, arranged according to the degree of closeness (P < 0.05); lymph nose metastasis rate was closely related with the size, infiltration depth, and gross type of the cancer (P < 0.05). In the same metastasis rate, the 5-year survivals rate decreased along with the increase of the amount of metastatic lymph nodes, and vice versa (chi(2) = 8.22, P = 0.004; chi(2) = 4.52, P = 0.033). CONCLUSION: The staging system based on metastatic lymph node rate is more predictable the number of metastatic lymph nodes for the prognosis of gastric cancer. PMID- 16313821 TI - [Residual stomach, duodenum, and continual jejunal interposition after subtotal gastrectomy]. AB - OBJECTIVE: To investigate the clinical effect of residual stomach, duodenum, and continual jejunal interposition on the patients of gastric cancer after subtotal gastrectomy. METHODS: Fifty-four patients with gastric cancer after subtotal gastrectomy were randomly divided into 2 groups: Group A (n = 26, receiving digestive tract reconstruction by manual end-to-side anastomosis of residual stomach and jejunum, end-to-side anastomosis of residual duodenum and jejunum, and side-to-side anastomosis of jejunum and jejunum, then the jejunum proximal to the stomach-jejunum anastomosis and the jejunum distal to the duodenum-jejunum anastomosis were ligated so as to form an integral continual jejunal interposition; and Group B (n = 28, receiving Bilroth digestive tract reconstruction. The operation time, body weight, prognosis nutrition index (PNI), and Visick score 3 and 6 months after the operation were observed. RESULTS: All patients recovered quickly and no complicating anastomosis leakage and obstruction was found. It took 53 +/- 9 minutes to finish the reconstruction in Group A, significantly shorter than that in Group B (57 +/- 6 minutes, t = 2.145, P = 0.037). The body weight and PNI of both groups decreased significantly 3 months after the operation in comparison with those before the operation (both P < 0.05). The body weight and PNI of Group A returned to the levels before operation. Although the body weight and PNI of Group B recovered to some extent 6 months after operation, they remained significantly lower than those before operation both P < 0.05). The Visick score 6 months after operation of Group A was superior to that of Group B (t = 2.1 P < 0.05). CONCLUSION: Residual stomach, duodenum, and continual jejunal interposition after subtotal gastrectomy helps overcome the difficulty in the procedure of digestive tract reconstruction and restore the physiological passage through duodenum, thus avoiding reflux and improving patients' quality of life. PMID- 16313822 TI - [Inhibitory effect of dendritic cells pulsed with MAGE-3 peptide on transplanted murine gastric cancer in mice]. AB - OBJECTIVE: To observe the inhibitory effect of MAGE-3 peptide pulsed DC on transplanted murine gastric cancer in 615 mice. METHODS: The CTL clones directed against MAGE-3 peptide were tested for the ability to lyse the gastric cancer cell line MFC which can express MAGE-3 antigen. In immunoprotection experiment, mice of the study group were immunized with MAGE-3 peptide pulsed DC (DC/MAGE-3) at the dosage of 1 x 10(6) on d0 and d7 by sc inoculation, mice of control groups were immunized with influenza virus peptide pulsed DC (DC/Nup) or unpulsed DC at the same dosage on days as the DC/MAGE-3 group. On d14, all the mice were challenged with sc injections of 5 x 10(5) MFC gastric cancer cells. In immunotherapy experiment, all the mice were sc injected 5 x 10(5) MFC gastric cancer cells on d0, and on d3, d10 mice of each groups were sc inoculated with DC/MAGE-3, DC/Nup or unpulsed DC at the dosage of 1 x 10(6) respectively. All mice were monitored closely with respect to tumor growth and survival times. RESULTS: The CTL clone induce by MAGE-3 peptide could lyse the MFC cells efficiently. Immunization of mice with DC pulsed with MAGE-3 generated partial protective immunity against MFC tumor, as well as significant inhibition of tumor growth in a 3-day tumor model. CONCLUSION: The tumor vaccine with DCs pulsed with MAGE-3 peptide possesses the ability to stimulate tumor specific CTL activity and to establish antitumor immunity when administered in vivo. PMID- 16313823 TI - [Refined mapping of loss of heterzygosity on 10q23-24 in sporadic colorectal carcinoma]. AB - OBJECTIVE: Loss of heterozygosity (LOH) of tumor suppressor gene (TSG) is believed to play a key role in carcinogenesis of colorectal cancer (CRC). In this study, we performed refined mapping of LOH on 10q23-24 region and screened the candidate TSG related to sporadic CRC. METHODS: Seven fluorescent labeled polymorphic markers (encompassing D10S185 locus) were analyzed in 83 cases of colorectal carcinoma and normal tissues by PCR. PCR products were eletrophoresed on an ABI 377 DNA sequencer. GeneScan 3.1 and Genotyper 2.1 softwares were used for LOH scanning and analysis. Comparison between LOH frequency and clinicopathological factors were performed by Chi-square test. RESULTS: The average LOH frequency was 36.11%. The highest frequency of LOH (D10S583 locus) and the lowest (D10S205 locus) were 54.84% and 21.3%, respectively. Two obviously high LOH regions were detected: one between D10S583 locus and D10S185 locus (about 0.9 cm, 10q23.33); another flanked by D10S1709 and D10S1265 locus (about 1.5 cm, 10q24.2-24.31). Furthermore, significant difference was observed between LOH frequency and Dukes stages only on D10S1265 locus. CONCLUSION: Two obviously high frequency LOH regions, 10q23.33 and 10q24.2-24.31, were detected in sporadic CRC. Besides PTEN gene, the above two regions may harbor candidate TSG involved in development and progression of sporadic CRC. PMID- 16313824 TI - [Proteome study of colorectal cancer genesis and hepatic metastasis]. AB - OBJECTIVE: To study differential proteins and their biological functions associated with colorectal cancer genesis and hepatic metastasis by proteomics and molecular biology techniques. METHODS: Isoelectric focusing/SDS acrylamide gel two-dimensional electrophoresis was used to analyse the expression of differential proteins from normal colorectal mucosa, primary cancer lesion and hepatic metastasis region. Liquid chromatography/mass spectrometry (LC-MS/MS) was used to identify the differential proteins. Transfection of colorectal cancer lovo cells was performed with the differential protein cDNA, and the changes of cell biological behavior was observed. RESULTS: Significant difference in protein expression was found on two-dimensional electrophoresis. Thirteen differential protein spots were analysed and identified. Human carbonic anhydrase II was detected in normal colorectal mucosa but not in primary cancer lesion and hepatic metastasisnegion. Phosphoglycerate kinase 1, fumarate hydratase and aldolase A were expressed in primary cancer. Expression of homo sapiens arginase and homo sapiens glutathione S-transferase A3 was found in hepatic metastasisnegion, but not in primary cancer lesion. After transfection with human carbonic anhydrase II cDNA, the lovo cells changed obviously with reduction in invasiveness, chemotaxy motor ability and tolerance. CONCLUSION: Differential expression of proteins was found between colorectal cancer genesis and hepatic metastasisnegion. No carbonic anhydrase II expression and enhanced expression of sapiens arginase and sapiens glutathione S-transferase A3 are related with biological behavior of colorectal cancer cell and facilitate hepatic metastasis. PMID- 16313826 TI - [Genetic polymorphisms of apoptosis-associated genes FAS and FASL and risk of colorectal cancer]. AB - OBJECTIVE: The FAS and FASL system plays a key role in regulating apoptotic cell death and corruption of this signaling pathway has been shown to participate in tumorigenesis. We previously have shown that the FAS-1377G/A and FASL-844T/C polymorphisms are associated with esophageal cancer. This case-control study was to examine the contribution of the polymorphisms to susceptibility of colorectal cancer. METHODS: PCR-RFLP method was used to determine the genotypes of FAS 1377G/A and FASL-844T/C in 382 patients with colorectal cancer and 648 controls. The adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression model. RESULTS: The subjects with the AA genotype of FAS-1377G/A and CC genotype of FASL-844T/C had an increased risk for colorectal cancer compared with those with the FAS-1377GG genotype (adjusted OR, 1.65; 95% CI, 1.14-2.38) and FASL-844TT genotype (adjusted OR, 1.76; 95% CI, 1.37-2.28). Furthermore, the effect of FAS and FASL polymorphisms on risk of colorectal cancer displayed a significantly gene-gene interaction (likelihood ratio test, P = 0.002). CONCLUSION: These findings suggest that the FAS-1377G/A and FASL-844T/C polymorphisms may be genetic susceptibility factors for colorectal cancer among Chinese population. PMID- 16313827 TI - [Expression of dihydropyrimidine dehydrogenase in human colorectal carcinoma and its clinical implications]. AB - OBJECTIVE: The study was to explore the dihydropyrimidine dehydrogenase (DPD) expression level in human colorectal carcinoma and its clinical implications. METHODS: Fifty-three patients with colorectal carcinoma were detected by immunohistochemical stain. These patients had undergone radical surgical treatment in the First Hospital of Xi'an Jiaotong University and had been followed up for 5 years after the operation. cases Twenty-two had received 5 fluorouracil-based adjuvant chemotherapy. RESULTS: DPD expression was predominantly observed in the cytoplasm of the tumor cells, and also partly in the nucleus. The positive rate of DPD expression was 73.58% (39/53). DPD expression in patients with different histopathology was obviously different (P < 0.05). There was no significant correlation between the expression of DPD and the efficacy of chemotherapy. By Kaplan-Meier methods, the survival patients with of negative DPD expression was longer than those with positive DPD expression, and then five-year survival rates were 12.97% and 42.86%, respectively (P < 0.05). The prognosis of patients with positive DPD expression was significantly poorer outcome than that of patients with negative DPD expression. CONCLUSION: These findings suggest that DPD expression might be one of the important prognostic parameters for colorectal cancer patients. PMID- 16313838 TI - Persistent efficacy of live attenuated hepatitis A vaccine (H2-strain) after a mass vaccination program. AB - BACKGROUND: Live attenuated hepatitis A vaccine (H2 strain) is widely applied in prevention of hepatitis A epidemic in China and other countries now. It is essential to observe and confirm the vaccine immune efficacy, population antibody level and its persistent efficacy after mass immunization. METHODS: A total of 220 children with negative anti-HAV antibody (aged 1 - 3 years) were taken for follow-up assay to observe seroconversion and geometric mean titre (GMT) level 2 months, 12 months, 6 years, and 10 years after inoculation. Another survey sampled from subjects of different age groups (3, 6, 9, 15, 18, 25 and 35 years) to compare anti-HA antibody positive rate before and after inoculation performed 10 years previously. Epidemiological observations were taken for 10 years to evaluate the relationship between vaccine coverage and hepatitis A morbidity. Serum antibody to HAV was detected by enzyme linked immunoassay (ELISA, calibrated by WHO international reference) and ABBOTT Axsym HAVAB microparticle enzyme immunoassay. RESULTS: Seroconversion in follow-up assay 2 months and 10 years after inoculation was 98.6% and 80.2% respectively. For children, the vaccination anti-HA antibody positive rates were significantly different before and after 10 years, 7.69% cf 70.45% (aged 3 years) and 52.58% cf 71.78% (aged 18 years). When vaccine coverage rose from 57% to 74%, there were no any HA epidemics. When vaccine coverage reached 85%, there were no any HA cases. With vaccine coverage between 85% and 91%, there were no any HA cases in cohorts from the age of 1 year to 15 years during the 10 years. CONCLUSIONS: Live attenuated hepatitis A vaccine has an obvious long-term effectiveness in prevention and control of HA epidemics through mass vaccination. PMID- 16313839 TI - Isolation and culture of adult Sertoli cells and their effects on the function of co-cultured allogeneic islets in vitro. AB - BACKGROUND: Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro. METHODS: Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro. RESULTS: In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01). CONCLUSIONS: A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation. PMID- 16313840 TI - Outcome of pancreaticoduodenectomy with extended retroperitoneal lymphadenectomy for adenocarcinoma of the head of the pancreas. AB - BACKGROUND: Nowadays, there is a remarkable rise in resectability rate of periampullary adenocarcinoma and the mortality and morbidity of the pancreaticoduodenectomy procedure have been reduced remarkably, while the 5 year survival rates of patients with carcinoma of the head of the pancreas are still below 25%. We conducted this retrospective study to evaluate the clinical outcome of radical pancreaticoduodenectomy with extended retroperitoneal lymphadenectomy as a surgical therapy for adenocarcinoma of the head of the pancreas. METHODS: Twenty cases with adenocarcinoma of the head of the pancreas were treated by standard pancreaticoduodenectomy (removing only the peripancreatic lymph nodes en bloc with the tumour) from 1994 to 1997, and 46 cases with the same disease underwent extended retroperitoneal lymphadenectomy associated with standard pancreaticoduodenectomy from 1998 to 2002. The patients for whom there were insufficient follow-up data, or who had received postoperative adjuvant therapy, were excluded from the analysis. Clinical and pathological parameters of both groups were reviewed. The postoperative morbidity, mortality and survival data were compared statistically. RESULTS: Demographic and histopathological characteristics were similar in the two groups of patients. Performance of the extended lymphadenectomy lengthened the procedure. The mean total number of lymph nodes resected was significantly higher in the radical group (P < 0.05). Of the 46 cases in the radical group, 26% (12/46) had metastatic adenocarcinoma in the resected retroperitoneal lymph nodes. There was one perioperative death in the standard group, and two in the radical group. Postoperative diarrhoea and lymphatic leakage were only observed in the radical group. Transfusion requirements and postoperative morbidity rates did not differ between the two groups. The 1-, 2- and 3-year survival rates were 63%, 32% and 21% respectively in the standard group, and 66%, 38% and 21% in the radical group. No statistically significant difference was found between the groups. When subgroups of node positive patients were analysed, the 1-, 2- and 3-year survival rates were 42%, 17% and 8% respectively in the standard group, and 65%, 32% and 16% in the radical group. Better survival was observed in the first 2 years after operation in the radical group, but no survival differences were seen after 2 years post operation. CONCLUSIONS: The addition of an extended lymphadenectomy to a pancreaticoduodenectomy did not significantly increase morbidity rates, but was associated with an early survival advantage. PMID- 16313841 TI - Polymorphisms in tumor necrosis factor genes and susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in a population of high incidence region of North China. AB - BACKGROUND: We investigated the possible association of the functional polymorphisms in the tumor necrosis factor (TNF) genes with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA). METHODS: The TNF-alpha-308G/A and TNF-beta+252G/A single nucleotide polymorphisms (SNPs) were genotyped using polymerase-chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, in 555 cancer patients (291 ESCC and 264 GCA) and 437 healthy controls in a high incidence region of North China. RESULTS: Among healthy controls, frequencies of the TNF-alpha 1/1, 1/2 and 2/2 genotypes were 89.4%, 9.2% and 1.4% respectively, while frequencies of the TNF-beta B1/B1, B1/B2 and B2/B2 genotypes were 12.6%, 32.3% and 55.1%, respectively. No significant difference was found in the overall genotype and allelotype distribution of the TNF-alpha-308G/A and TNF-beta+252G/A SNPs among cancer patients and controls. However, both the B1/B1 genotype and B1/B2 genotype significantly increased the risk of developing ESCC [the age and gender adjusted odds ratio (OR) = 2.04 and 1.91, 95% confidence interval (CI) = 1.04 - 4.43 and 1.14 - 2.60, respectively] and GCA (the age and gender adjusted OR = 2.68 and 2.64, 95% CI = 1.14 - 6.29 and 1.47 - 4.72, respectively) in individuals with negative family history of UGIC, in comparison with the B2/B2 genotype. When the two TNF polymorphisms were combined and analyzed, individuals with the TNF-beta B1/B2 and TNF-alpha 1/2 or 2/2 genotypes significantly reduced the risk of developing ESCC and GCA, in comparison with those harboring the TNF-beta B2/B2 and TNF-alpha 1/1 genotypes (the age and gender adjusted OR = 0.37 and 0.34, 95% CI = 0.15 - 0.92 and 0.13 - 0.90, respectively). CONCLUSIONS: Therefore, the TNF alpha-308G/A and TNF-beta+252G/A genotyping may be used as a stratification markers to predicate the risk of ESCC and GCA development in North China. PMID- 16313842 TI - Diagnosis and treatment of bacterial pneumonia in liver transplantation recipients: report of 33 cases. AB - BACKGROUND: Bacterial pneumonia in the recipients of liver transplantation (LTX) is a common postoperative complication influencing the prognosis greatly. In this article, the diagnosis and treatment of bacterial pneumonia in 33 LTX recipients are reported. METHODS: From February 1999 to January 2003, a total of 103 patients underwent allogeneic LTX at our center; afterwards, a retrospective analysis was made on their postoperative clinical manifestations, including symptoms (expectoration, panting and fever), sign (rale), results of laboratory examinations (white blood cell count and sputum culture of tracheal secretions or pleural fluid culture), and chest X-ray films. The following data of the pneumonia and non-pneumonia groups were collected, and the rank sum test (SPSS 11.0, Wilcoxon's method) was used to analyze the duration of postoperative respirator utilization and the volume of pleural effusion through pleurocentesis or pleural drainage. RESULTS: In the 103 patients, 33 experienced 53 episodes of bacterial pneumonia during their hospital stay after transplantation, 14 of them (42.42%) had more than three manifestations of the seven mentioned above. The pathogens causing bacterial pneumonia included Pseudomonas aeruginosa (17.48%), Klebsiella pneumoniae (15.53%), Acinetobacter baumannii (10.68%), and Staphylococcus aureus (7.77%). Amilkacin, tienam, ciprofloxacin, vancomycin, etc. were the antibiotics of choice against those bacteria. Acute rejection occurred during the treatment of bacterial pneumonia in 16 patients, and 5 of them died. Wilcoxon's rank sum test of the data indicated that the pneumonia group had longer duration of postoperative ventilator treatment and larger volume of pleural effusion than the non-pneumonia group (P < 0.05). CONCLUSIONS: The clinical manifestations of pneumonia after LTX might be atypical, and special attention should be paid to the respiratory symptoms and signs within 2 months after LTX. Whenever the diagnosis of bacterial pneumonia is confirmed, consideration should be given to reasonable use of antibiotics and regulation of immunity in addition to other routine therapies. PMID- 16313843 TI - Changes of chymase, angiotensin converting enzyme and angiotensin II type 1 receptor expressions in the hamster heart during the development of heart failure. AB - BACKGROUND: Little is known about the role of dual angiotensin II forming pathways during heart failure. In the present study, the changes of chymase and angiotensin converting enzyme (ACE) expressions in the failing hearts of hamsters were analysed. METHODS: Heart failure was induced by ligation of left anterior descending branch of the coronary artery. Chymase, ACE and angiotensin II type 1 receptor (AT1R) mRNA levels were analysed by reverse transcription polymerase chain reaction (RT-PCR). The activities of chymase and ACE were determined by radioimmunoassay (RIA). Myocardial collagen fibre analysis was performed under optical microscope. RESULTS: Left ventricular systolic pressure (LVSP) and maximum left ventricular developed pressure increase rate (dp/dtmax, mmHg/s) gradually moved lower at 2, 3, 4 and 8 weeks after operation. On the other hand, left ventricular end-diastolic pressure (LVEDP) increased gradually after operation. Compared with the control group (3.55 +/- 0.06, 4.79 +/- 0.70), the heart weight/body weight ratio in operation group had increased significantly at 4 weeks and 8 weeks (4.28 +/- 0.43, 6.17 +/- 0.73) (P < 0.01). Collagen staining showed that the quantity of myocardial collagen fibre increased significantly in the operation group. RT-PCR showed that the chymase mRNA level in the operation group was consistently greater than that in the control group. AT1R mRNA level was also increased significantly at 3 weeks and 4 weeks, both being 1.3 times that of the control group (P < 0.01), whereas ACE mRNA level was not changed. Higher activity of chymase was detected in operation group, being 4, 8, 13 and 19 times that of the control group at 2, 3, 4 and 8 weeks (P < 0.01), respectively. ACE activity was also significantly higher at the same time, being 7, 10, 10 and 3.5 times that of the control (P < 0.01). Angiotensin II (Ang II) level in operation group increased significantly, being 2.5, 2.7, 3.5 and 2 times that of the control group at 2, 3, 4 and 8 weeks, respectively (P < 0.01). CONCLUSIONS: A dual Ang II forming pathway from both ACE and chymase in the hamster hearts plays an important role during the development of heart failure. At the decompensatory stage, the reduction of AngII level may be associated with the decrease of ACE activity. PMID- 16313844 TI - The role of NF-kappaB activation in lipopolysaccharide induced keratitis in rats. AB - BACKGROUND: Nuclear factor-kappa B (NF-kappaB) is elevated in regulating transcription of many cytokines and inflammatory mediators. The purpose of this study was to investigate the activation and the significance NF-kappaB in lipopolysaccharide (LPS) induced keratitis. METHODS: LPS induced keratitis model was based on Wistar rats. At 0.5, 1, 3, 6, 12, 24 or 72 hours after LPS treatment, the rat corneas were observed with a slit lamp microscope, then the rats were sacrificed and their corneas were excised for routine histological analysis. The expression of NF-kappaB was detected with immunohistochemical staining. The change of tumour necrosis factors-alpha (TNF-alpha) mRNA expression was identified by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Histological findings demonstrated that LPS treated corneas showed significant changes in corneal structure. Corneal edema, pronounced inflammatory cells infiltration and inordinate collagen fibres were observed. Immunohistochemical results showed that the expression of NF-kappaB and its activation obviously increased after LPS treatment compared with the normal group and control group. Positive cells could be observed at 0.5 hour and peak expression of NF-kappaB was observed between 3 hours and 12 hours after infection, but returned to or approached normal level by 72 hours. RT-PCR showed that the level of TNF-alpha mRNA began to increase 0.5 hour after LPS treatment, peaked at 6 hours and then subsided by 72 hours. NF-kappaB had a positive correlation with the expression of TNF-alpha mRNA (r = 0.964, P < 0.01), both NF kappaB and TNF-alpha had a strong positive correlation with the degree of inflammatory response in LPS treated corneas (r = 0.929, P < 0.01; r = 0.587, P < 0.05, respectively). CONCLUSIONS: The activation of NF-kappaB was increased in LPS treated corneas and was elevated in LPS induced keratitis by promoting overexpression of TNF-alpha mRNA. NF-kappaB may play an important role in the pathogenesis of LPS-associated keratitis in rats. PMID- 16313845 TI - Up regulation of interleukin-8 expressions induced by mast cell tryptase via protease activated receptor-2 in endothelial cell line. AB - BACKGROUND: Protease activated receptor-2 is cleaved and activated by trypsin or mast cell tryptase and may play an important role in inflammation. However, it is unknown whether PAR-2 can mediate tryptase-induced inflammatory reaction. This study was conduct to investigate whether PAR-2 could be the activated by mast cell tryptase and medicated the tryptase induced interleukin-8 expression in endothelial cells. METHODS: Protease activated receptor-2 expression was found in endothelial cell lines ECV304 cell by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Interleukin-8 stimulated by purified human mast cell tryptase was determined by RT-PCR and enzyme linked immunosorbent assay (ELISA). Data were analysed by the S-N-K one-way ANOVA test. RESULTS: The present study shows that mRNA and protein of protease activated receptor-2 could be expressed in ECV304 cells, and tryptase upregulated the expression levels of both interleukin-8 mRNA and protein. The increased expression of interleukin-8 was inhibited by an antiprotease activated receptor-2 monoclonal antibody, SAM11. An additional band was observed by Western blotting after the incubation of ECV304 cells with tryptase for 2 hours, which suggested that protease activated receptor-2 was activated. CONCLUSION: Protease activated receptor-2 can mediate the mast cell tryptase stimulated expression of interleukin-8 in ECV304 cell. PMID- 16313846 TI - RNA interference inhibits expression of vascular endothelial growth factor (VEGF) in human retinal pigment epithelial cells. PMID- 16313847 TI - Distinct serumal proteomic patterns between ascending and descending types of loco-regionally advanced nasopharyngeal carcinoma assessed by surface enhanced laser desorption ionization and artificial neural network analyses. PMID- 16313848 TI - Effect of different iodine intake on the prevalence of hypothyroidism in 3 counties in China. PMID- 16313849 TI - Comparison of airway resistance measurements by the forced oscillation technique and the interrupter technique for detecting chronic obstructive pulmonary disease in elderly patients. PMID- 16313850 TI - Improving capacity building by implementing comprehensive eye care project, experiences from Kuandian County, China. PMID- 16313851 TI - Recurrent malignant carotid body tumor: report of one case and review of the literature. PMID- 16313852 TI - Magnetic resonance imaging findings in Lhermitte-Duclos disease: reports of three cases. PMID- 16313853 TI - Pay for performance: an alternative to Medicare fee schedules? PMID- 16313854 TI - Integration and standardization within the Mayo Foundation Laboratories: the centralized laboratory purchasing group. AB - Integration and standardization of laboratories throughout a medical system can increase the efficiency and effectiveness of laboratory operations. This task is challenging in most health-care systems, as no central governance exists to compel laboratories to standardize and integrate. We describe the initial collaborative efforts to integrate and standardize the laboratories of the Mayo Foundation, which includes more than 60 laboratories of different sizes in diverse locations. The goals and objectives of the group formed to develop this initiative--the Centralized Laboratory Purchasing Group--its origin, and lessons learned are described. Similar initiatives by other academic medical centers and community health-care systems to integrate and standardize their laboratories are discussed. Successful standardization and integration increases the value of the laboratory to the larger health-care system by demonstrating accountability, efficiency, and effectiveness, and can result in considerable cost savings to the entire health-care system. PMID- 16313855 TI - Experience of implementing a quality management system in an Ontario hospital laboratory. AB - This article describes the process of building and implementing a quality management system in the Department of Laboratory Medicine at St. Michael's Hospital in Toronto. This was done in part to fulfill the requirements of the Ontario Laboratory Accreditation program. During the process, we revised and created new procedure manuals and documents through the use of focus groups and inter-departmental committees. The entire project took approximately two-and-a half years to complete and required teamwork, personal commitment, and professional sacrifices by key personnel. PMID- 16313856 TI - Maintaining civility in the laboratory. AB - Uncivil behavior in the laboratory can be crippling, damaging the morale of the department and overall attitude of employees. In this article, the author looks at data that indicates the importance of curtailing incivility as soon as possible and offers suggested ways for managers to deal with uncivil behavior. PMID- 16313857 TI - TB... or not TB?--part two. PMID- 16313859 TI - Critical quality management tools. PMID- 16313858 TI - Informing patients about violations of confidentiality. PMID- 16313860 TI - Management of chronic hepatitis C virus in patients with HIV. AB - The life expectancy of HIV seropositive persons is approaching the life expectancy of those who are uninfected with HIV. Hepatitis C virus (HCV) infection has emerged as a worldwide epidemic. Given the similar transmission route between HCV and HIV, there has been an explosion in the number of individuals infected with both viruses. Because of the successful introduction of antiretroviral therapy, patients are more susceptible to new opportunistic infections such as HCV. HCV leads to a more rapid progression to end-stage liver disease in patients with HIV, and the morbidity and mortality related to HCV in co-infected patients is on the rise. Therefore, it has become imperative to treat both HIV and HCV in co-infected patients. The primary goal of HCV therapy is permanent eradication of the virus. Secondary goals include reduction in hepatic fibrosis progression, development of decompensated cirrhosis, and hepatocellular carcinoma. Early studies using standard interferon-alfa for the treatment of HCV in co-infected individuals were discouraging, as poor outcomes, high discontinuation rates, and severe adverse events were observed. The current standard of care for treatment of HCV is pegylated-interferon and ribavirin. New studies have recently demonstrated a higher sustained virologic response rate and a better adverse event profile than previously reported in co-infected patients. As a result, we recommend considering all co-infected patients for HCV therapy while watching closely for unique treatment-related toxicities. The treatment of HCV in co-infected patients should be a high priority for all providers. PMID- 16313861 TI - Hepatorenal syndrome. AB - Hepatorenal syndrome (HRS) is defined as functional renal failure that develops in patients with advanced liver disease. HRS may be either slowly or rapidly progressive (type I and II HRS, respectively). Untreated HRS carries a high mortality. Liver transplantation is the best available treatment for HRS. However, all patients with HRS are not suitable candidates for transplantation. Moreover, an organ is often not available in a timely manner in those who are candidates for transplantation. Treatment with vasoconstrictors (terlipressin, octreotide, and midodrine) and plasma expansion with albumin is beneficial and serves as a bridge to transplantation in such cases. The vasopressin analog, terlipressin, produces a sustained reversal of HRS in about 57% to 78% of the patients. The benefits of terlipressin are seen mainly in those who are also receiving albumin simultaneously. In those who improve, recurrence of HRS is reported to be relatively uncommon in the short and intermediate term. In the United States, terlipressin is not available, and octreotide and midodrine are often used for the medical management of HRS. Unfortunately, there are only limited uncontrolled data to support the use of these drugs for HRS. In those who respond to octreotide and midodrine, the subsequent placement of a transjugular intrahepatic portasystemic shunt (TIPS) has been shown to produce a sustained improvement in renal function. TIPS alone also improves renal functions in selected patients with HRS. The exact role of TIPS in HRS needs further evaluation, as patients with HRS are particularly at risk for complications such as encephalopathy and liver failure. Molecular adsorbent recirculating system (MARS) is an albumin-based dialysis system that has a promising role in the treatment of HRS and liver failure. MARS is a very expensive form of treatment, and further clinical trials are needed to establish its utility. Development of HRS can be prevented by adding albumin to the antibiotic regimen to treat spontaneous bacterial peritonitis and through pentoxifylline administration to the patients with acute alcoholic hepatitis. PMID- 16313862 TI - Hepatopulmonary syndrome. AB - The hepatopulmonary syndrome (HPS) is an important and often under-recognized vascular complication of cirrhosis and portal hypertension characterized by pulmonary vascular dilatation, which results in hypoxemia. This syndrome is identified in as many as 20% of patients who are evaluated for orthotopic liver transplantation (OLT), and it has recently been found to increase mortality in affected patients, particularly when hypoxemia is severe. Currently, OLT is the only therapy established to reverse intrapulmonary vasodilatation, although postoperative mortality is increased in patients with severe hypoxemia. No randomized controlled trials of pharmacologic therapies have been undertaken, but supplemental oxygen improves oxygenation. Data derived from case reports, small studies, and experimental models suggest that pharmacologic therapies may be effective. In cirrhotic patients with HPS, particularly those with moderate hypoxemia (PaO2 < 60 mmHg), OLT should be considered prior to the development of severe deoxygenation. Supplemental oxygen should be given to patients with a PaO2 < 60 mmHg or those with exercise oxygen desaturation. For those patients with mild hypoxemia or those who are not OLT candidates, a trial of pharmacologic treatment may be considered. PMID- 16313863 TI - Hepatocellular carcinoma: current treatment strategies. AB - Hepatocellular carcinoma (HCC) is an increasingly prevalent clinical problem. The presence of cirrhosis in the majority of patients makes treatment difficult because both the stage of the tumor and the stage of cirrhosis must be taken into account. This is compounded by the difficulty in diagnosing HCC in the early stages, where treatment is most effective, and the lack of a globally accepted treatment policy. Liver transplantation and liver resection are the optimal treatments, with resection being preferred in patients with small lesions, clinically well-preserved liver function, and absence of portal hypertension. Patients unsuitable for these procedures, due to localized but large tumor bulk, are only treatable by ablative and palliative therapies. Ablation involves either thermal (preferably radiofrequency ablation) or chemical methods, with the choice of method being dependent on both the size and placement of the tumor and the operator. Ablation may also be used as a bridge to transplantation in centers where significant waiting times are anticipated. Tumors that are too large in size or number to ablate are treated with transarterial chemoembolization, involving the distribution of chemotherapeutic agents and the blocking of the blood supply to the tumor; this is not considered a curative therapy. Combination therapies may also be used. These treatment options need further evaluation for determination of the optimal course of therapy for individual patients. PMID- 16313864 TI - Hepatic Wilson's disease: initial treatment and long-term management. AB - This article is based on the experience of 320 patients with Wilson's disease who were seen between the years 1954 and 2000. These patients were seen at The Boston City Hospital, 1954 thru 1955, University College Hospital, London,1955 thru 1957; Addenbrooke's Hospital, Cambridge, 1967 thru 1987, and The Middlesex Hospital London, 1988 thru 2000. Wilson's disease is not strictly a gastroenterologic disease but a genetically determined metabolic disease that is mediated by a failure of copper excretion through the bile. The mutation carried on chromosome 13q14.3: it involves a copper-carrying ATPase (ATPase 7B); more than 250 mutations are now known. The first organ to be affected is the liver, then many other tissues, principally the brain but also the eyes, the kidneys, the bone marrow, and the osteoskeletal system. It is with the hepatic form of the disease that this article is concerned. The hepatic illness may be acute, subacute or chronic; it may be progressive or, apparently, self-limiting. In 10% of patients hemolysis may also be found which can later lead to the formation of pigment gallstones. The management of liver disease is not considered in this article, which is strictly confined to the therapeutic options available for the elimination of copper and the long-term welfare of the patient. It must be remembered that all close relatives of the patient must be screened for the presymptomatic stage of the disease so, if they are found to be homozygous carriers for the mutation, they can be started on preventive treatment. PMID- 16313865 TI - Management of the cirrhotic patient that needs surgery. AB - Conditions that necessitate surgery frequently arise in patients with chronic liver disease and cirrhosis. Because cirrhosis has the ability to cause physiologic derangements in every organ system in the body, clinicians face significant challenges in preoperative preparation of the patient with cirrhosis in order to decrease postoperative morbidity and mortality. Emergent operations add an extra dimension of complexity to the clinical picture, due to limited preoperative time to prepare the patient with cirrhosis for surgery. In cases of severely decompensated cirrhosis, clinicians should have in their armamentarium possible alternatives to surgery that can be used to temporize the emergent nature of the disease and improve patient outcomes. The classification of cirrhotic liver disease by Child and Turcotte was initially utilized to predict mortality in patients undergoing surgically placed shunts for portal hypertensive bleeding. Subsequent studies have pointed to the fact that other general and thoracic surgery procedures can be assigned predicted mortality rates according to a similar classification scheme, the modified Child-Pugh score. Patients with cirrhosis facing surgery should undergo a careful history and physical examination and should be accurately placed into a designated Child-Pugh category. Because the modified Child-Pugh class is the most reliable determinant of postoperative morbidity and mortality, every attempt should be made to upgrade a patient's class in a favorable direction prior to surgery. Patients should be carefully evaluated for the presence of ascites and dietary alterations. In addition, medical management with diuretics should be employed to prevent postoperative ascites leak and possible infectious complications including bacterial peritonitis. Perhaps one of the most feared complications in the patient with cirrhosis facing surgery is hemorrhage. Because the liver is vital in maintenance of coagulation homeostasis, several pharmacologic adjuncts may be administered to correct any coagulopathy in the peri-operative period. Several diseases such as cholelithiasis and peptic ulcer disease are known to be more prevalent in the cirrhotic patient, and clinicians treating these diseases should have a thorough understanding of the pathophysiology of cirrhosis and portal hypertension. Patients with cirrhosis and portal hypertensive bleeding that are considered good surgical candidates (ie, Child-Pugh class A) may benefit from surgical portasystemic shunt in contrast to angiographically placed portacaval shunt (ie, transjugular intrahepatic portosystemic shunt ) due to the lack of durable patency and cost effectiveness in the latter. In patients with cirrhosis awaiting orthotopic liver transplantation, TIPS may be a lifesaving temporizing technique that is utilized as a bridge to transplantation. PMID- 16313866 TI - Autoimmune hepatitis. AB - Autoimmune hepatitis (AIH) is a progressive inflammatory hepatitis of unknown etiology that is responsive to immunosuppressive therapy. The diagnosis of AIH should be guided by the descriptive criteria and scoring system set forth by the International Autoimmune Hepatitis Group. Standard therapy is prednisone with or without azathioprine (AZA). Combination therapy with prednisone and AZA is preferred, as it allows treatment with lower individual doses of each drug and is thus associated with fewer side effects. Treatment goals include complete biochemical, clinical, and histologic remission. Treatment outcomes include complete remission with or without relapse, incomplete response, or treatment failure. Treatment withdrawal, once remission has been attained, may be associated with relapse of disease. Recurrent relapse may be addressed with long term, lower dose maintenance treatment with prednisone or AZA. Incomplete response is addressed by attempting high-dose immunosuppressive regimens or by considering investigational medical regimens. Treatment failure is also addressed by considering investigational medical regimens. Medication toxicity is addressed with dose reduction or drug discontinuation. Should the patient progress to decompensated cirrhosis, liver transplantation is an effective treatment for AIH. Calcium and vitamin D supplementation, a well-balanced diet, and exercise are advocated in patients with AIH. Cyclosporine, mycophenolate mofetil, and tacrolimus are promising agents among the new therapies for autoimmune hepatitis. Unfortunately, newer therapeutic agents have been studied in small numbers. Larger, controlled treatment trials are needed to expand the repertoire of therapeutics to treat patients with fewer side effects and to provide alternatives for patients who are refractory to conventional therapy. PMID- 16313867 TI - Cholangiocarcinoma. AB - Cholangiocarcinoma is an uncommon malignant tumor arising from biliary epithelium. The incidence increases with age and usually affects individuals in their sixth or seventh decade of life. Most patients clinically present with features of biliary obstruction. Although surgical resection offers the only hope for cure, the majority of patients are found to have unresectable disease on initial presentation and carry extremely grim prognosis. This has lead to an emphasis on the role of palliative care, with the relief of biliary obstruction being the primary goal in the management of these patients. Surgical bypass was once considered as the primary means of palliating biliary obstruction, but nonsurgical placement of biliary stents is associated with lower morbidity and mortality. Newer modalities such as photodynamic therapy, brachytherapy, and high intensity ultrasound therapy may result in improved survival and play a future role as an adjunctive therapy to surgical resection. PMID- 16313868 TI - Fulminant hepatic failure. AB - The rare but potentially devastating clinical syndrome of fulminant hepatic failure has as its components severe encephalopathy and finally cerebral edema, hemodynamic instability, renal failure, coagulopathy, profound metabolic disturbances and a particular susceptibility to bacterial and fungal infection. Despite advances in medical management, fulminant hepatic failure in its most severe form carries a high mortality rate unless urgent orthotopic liver transplantation is carried out. However, availability of cadaveric donor organs is limited and, due to the rapidly progressive clinical course in many cases, a substantial proportion of patients will die or develop contraindications to transplantation before the procedure can be performed. Consequently, recent interest has centred on living donor transplantation and the possibility of providing temporary liver support, either through auxiliary partial organ transplantation, extracorporeal perfusion or transplantation of hepatocytes, to allow time for either a liver graft to become available or native liver regeneration, on which spontaneous survival ultimately depends, to occur. PMID- 16313869 TI - Is Doppler investigation useful in ulcer bleeding? PMID- 16313872 TI - Pathogenesis and therapy for idiopathic dyspepsia. AB - Functional dyspepsia (FD) is one of the most common gastrointestinal disorders. This review summarizes recent progress in our understanding of the pathogenesis and therapy for FD. Although distinction among FD, irritable bowel syndrome, and reflux disease is difficult in population-based studies, separate entities can be recognized in patients who seek medical attention. The pathogenesis of FD remains unclear, but recent studies have demonstrated a role for acute gastrointestinal infection in triggering FD and in genetic polymorphisms of G-proteins in predisposing to FD. The role of abnormalities in gastric motor function, visceral hypersensitivity, and psychosocial factors in the pathophysiology of dyspeptic symptoms has been the topic of multiple studies. Treatment options for FD remain limited. Recent studies have focused on acid-suppressive drugs and on novel prokinetics. Progress in our understanding of the pathogenesis and pathophysiology of FD may lead to new or improved treatment modalities. Areas of major advances are the role of infection and genetic predisposition and studies on the role of abnormalities in gastric motility and sensitivity. PMID- 16313873 TI - Obesity in the United States: is there a quick fix? Pros and cons of bariatric surgery from the pediatric perspective. AB - Bariatric surgery has become a standard of care for the treatment of severely obese adults who meet National Institutes of Health criteria. Unfortunately, there is a need for weight loss surgery in adolescents. Criteria developed for adolescents are generally more restrictive than those for adults due to concerns about the ability of children to consent for life-altering surgery and the lack of long-term results. Several preliminary series have reported adolescent bariatric procedures with promising results. The two primary bariatric procedures performed on adolescents are Roux-en-Y gastric bypass (RYGB) and adjustable gastric banding (AGB). Each procedure has its advantages and disadvantages. RYGB has the longest follow-up results and is considered the "gold standard." AGB has promising results, is reversible, and can be performed with less morbidity and mortality. PMID- 16313874 TI - Obesity in the United States: is there a quick fix? Pros and cons of bariatric surgery from the adult perspective. AB - Over 60% of the American population is either overweight or obese. Conventional treatment through decreased caloric intake and increased physical activity has been minimally effective in achieving sustained weight loss. Pharmacotherapies are effective in inducing some weight loss; however, lost weight is regained once the medications are stopped. Surgical weight loss is the only treatment that has been demonstrated to achieve durable weight loss, with sustained weight loss for at least 14 years and improvements in or complete resolution of associated comorbidities and improved quality of life. Based on the National Institutes of Health 1991 Consensus Conference guidelines, 16 million Americans are currently potential candidates for weight loss surgery. Although this is an effective treatment for appropriate candidates in the short term, education on nutrition and increased physical activity is more likely to be effective over the long term. One hopes that current research efforts in obesity and public awareness will lead to a decrease in the prevalence of this disease. PMID- 16313875 TI - Celiac disease: is the atypical really typical? Summary of the recent National Institutes of Health Consensus Conference and latest advances. AB - Celiac disease (CD) is an immune-mediated enteropathy triggered by the ingestion of gluten in genetically susceptible individuals. Gluten is a protein component in wheat and other cereals, such as rye and barley. At present, the only available treatment is a strict gluten-free diet. Recent advances have increased our understanding of the molecular basis for this disorder. The past decade has seen new scientific developments in this disease and led to the formulation of new concepts of pathophysiology and clinical manifestations. There are several targets for new treatments. This article briefly summarizes the National Institutes of Health Consensus Statement and gives an overview of new findings in recent years and of future therapeutic options for CD. PMID- 16313877 TI - Evolving diagnostic modalities in inflammatory bowel disease. AB - Over the past several years, significant advances have been made in the diagnostic techniques used in the management of ulcerative colitis and Crohn's disease. These advances have occurred mainly in the area of gastrointestinal endoscopy and radiology. Capsule endoscopy and double-balloon endoscopy have permitted better visualization of the small bowel mucosa. Advanced imaging techniques, including chromoendoscopy, magnification endoscopy, confocal endomicroscopy, and spectroscopy, may aid in the diagnosis of colorectal neoplasia in patients with long-standing disease. Improved radiographic imaging techniques based on computed tomography and magnetic resonance imaging allow noninvasive means of evaluating the small bowel in patients with known or suspected Crohn's disease. Finally, positron emission tomography is an investigative tool for inflammatory bowel disease that may also aid in the detection of inflammation in these diseases. PMID- 16313878 TI - Advances in therapeutic approaches to ulcerative colitis and Crohn's disease. AB - Advances (from 2004 to 2006) in the use of conventional agents include the molecular mechanisms of action, which have implications for monitoring (azathioprine and thioguanosine triphosphate) and chemoprevention (mesalamine and peroxisome proliferator activated receptor gamma). Advances in biotherapy include new data on monoclonal antibodies (infliximab in ulcerative colitis, adalimumab, certolizumab pegol, fontolizumab, selective anti-adhesion molecules, and others), antisense oligonucleotides, the development of small molecules, and cell-gene therapy (including helminth ova, leukocytapheresis, stem cell transplantation, and probiotic intestinal mucosal delivery systems). However, management of inflammatory bowel disease is about more than drug therapy, dose, and timing. The goals remain induction of remission, limitation of side effects, modification of the pattern of disease, and avoidance of complications. With the cost and complexity of biotherapy, inflammatory bowel disease is emerging as a specific subspecialty. PMID- 16313879 TI - Positioning novel biologic, probiotic, and apheresis therapies for Crohn's disease and ulcerative colitis. AB - Traditional medications for inflammatory bowel disease are small molecule drugs, most of which were developed for use in other diseases before being found to be efficacious for the treatment of ulcerative colitis or Crohn's disease. Recently, several exciting alternative approaches to the medical treatment of inflammatory bowel disease have been developed. These include biologic, probiotic, and apheresis therapies that offer certain advantages over traditional drug therapy for inflammatory bowel disease. The purpose of this review is to assess the current state of knowledge about novel biologic, probiotic, and apheresis therapies and to analyze how best to incorporate these therapies into evolving management paradigms of inflammatory bowel disease. PMID- 16313880 TI - Postoperative management of ulcerative colitis and Crohn's disease. AB - Approximately 10% to 30% of patients with ulcerative colitis and up to 70% of patients with Crohn's disease will undergo surgery at some point during their lifetime. Although patients with ulcerative colitis are considered "cured" by surgery, patients who have undergone an ileal pouch anal anastomosis may develop pouchitis, cuffitis, pouch irritability, or even Crohn's disease. Various therapies have shown success, including probiotics, in the prevention of pouchitis onset or relapse. Crohn's disease historically recurs following surgery; prophylaxis against disease recurrence has been attempted with a variety of agents, with variable success. Innovative therapies holding promise for the future treatment or prevention of these conditions are under exploration. PMID- 16313881 TI - Hazard perception and driving experience among novice drivers. AB - The crash risk of novice drivers decreases rapidly during the first few months of post-licensing driving, indicating that some important safety-relevant traffic skills are learnt during this period. The present study tested the hypothesis that the risk decrease is related to improved hazard perception skills. A video based hazard perception/reaction test, measuring reaction times to 31 traffic scenes, was administered to three groups of drivers, having held a licence for 1, 5, and 9 months, respectively, and to a group of drivers who had held their licences for several years. Average reaction times tended to decrease with experience, but the decrease was not significant. There were some significant differences in the expected direction for individual test items, indicating a possible effect of experience. One half of the situations were presented together with a secondary task, in order to investigate effects of increased mental load on hazard perception. Male novice drivers had relatively longer reaction times with the secondary task, compared to both female novice drivers and male experienced drivers. It is concluded that hazard perception as tested here is probably only a minor factor in explaining the initial risk decrease among novice drivers. PMID- 16313882 TI - Rational design of complementary peptides to the betaAmyloid 29-42 fusion peptide: an application of PepDesign. AB - Peptides in solution currently exist under several conformations; an equilibrium which varies with solvent polarity. Despite or because of this structure versatility, peptides can be selective biological tools: they can adapt to a target, vary conformation with solvents and so on. These capacities are crucial for cargo carriers. One promising way of using peptides in biotechnologies is to decipher their medium-sequence-structure-function relationships and one approach is molecular modelling. Only few "in silico" methods of peptide design are described in the literature. Most are used in support of experimental screening of peptide libraries. However, the way they are made does not teach us much for future researches. In this paper, we describe an "in silico" method (PepDesign) which starts by analysing the native interaction of a peptide with a target molecule in order to define which points are important. From there, a modelling protocol for the design of 'better' peptides is set. The PepDesign procedure calculates new peptides fulfilling the hypothesis, tests the conformational space of these peptides in interaction with the target by angular dynamics and goes up to the selection of the best peptide based on the analysis of complex structure properties. Experimental biological assays are finally used to test the selected peptides, hence to validate the approach. Applications of PepDesign are wide because the procedure will remain similar irrespective of the target which can be a protein, a drug or a nucleic acid. In this paper, we describe the design of peptides which binds to the fusogenic helical form of the C-terminal domain of the Abeta peptide (Abeta29-42). PMID- 16313883 TI - Evaluation of therapist-supported parent-implemented CBT for anxiety disorders in rural children. AB - Supplementing bibliotherapy with therapist-client communication has been shown to be an effective way of providing services to under-resourced and isolated communities. The current study examined the efficacy of supplementing bibliotherapy for child anxiety disorders with therapist-initiated telephone or email sessions, or with client-initiated contact in a randomised trial using a waitlist control. Participants were 100 anxiety-disordered children and their parents from rural and remote communities. All treatment conditions resulted in improvement on self-report measures and clinician rated severity. Telephone sessions produced superior outcomes with 79% of children being anxiety disorder free post-treatment compared with 33% of email and 31% of client-initiated participants. The results suggest that therapist supplemented bibliotherapy could provide an efficacious treatment option for families isolated from traditional treatment services. PMID- 16313884 TI - Elicitation of neutralizing antibodies by intranasal administration of recombinant vesicular stomatitis virus expressing human immunodeficiency virus type 1 gp120. AB - Recombinant viral vectors are useful tools for AIDS vaccine development. However, expression of HIV-1 envelope genes using viral vectors has not been successful in the induction of potent neutralizing antibodies in vivo. We took advantage of the strong immunogenicity of vesicular stomatitis virus (VSV)-based vector and expressed HIV-1 HXB2 gp120 gene in the recombinant VSV. Our results showed that HIV-1 gp120 protein expressed by the recombinant VSV retained the native conformation of the protein to some degree and was recognized by two well characterized broad anti-HIV-1 neutralizing monoclonal antibodies b12, 2G12. We further showed that only one time intranasal immunization with the recombinant VSV led to production of anti-HIV-1 anti-sera in mice. In addition, we found that the anti-sera had the ability to neutralize not only HXB2 envelope-pseudotyped HIV-1 viruses but also HIV-1 pseudotyped viruses with JRFL envelopes. These results suggest that HIV-1 gp120 expressed by the recombinant VSV, in combination with the route of intranasal administration, is an effective strategy to evaluate the immunogenicity of HIV-1 envelope protein and its variants in mice. PMID- 16313885 TI - Crystal structure and some properties of a major house dust mite allergen, Derf 2. AB - Pyroglyphid house dust mites are a major source of allergens in house dust. Mite allergens sensitize and induce asthma, rhinitis, and eczema in a large portion of patients with allergic diseases. Here, the crystal structure of a major mite allergen, Derf 2, derived from Dermatophagoides farinae was solved by single isomorphous replacement method with anomalous scattering (SIRAS) at 2.1A resolution. The present study also demonstrated that the conformation of the allergen was critical in the determination of Th1/Th2 shift based on physicochemical and immunological analyses. This indicates that rigidly folded and singly dispersed structure is essentially required for the generation of Th2 type cells by the allergen, while conformational variant protein leads to Th1 skewing, irrespective of the same amino acid sequence. This structure/function relationship may allow us to develop a novel strategy for hyposensitization therapy in patients with allergic diseases triggered by house dust mite allergens. PMID- 16313886 TI - The p53 oncoprotein is a substrate for tissue transglutaminase kinase activity. AB - Increased expression and activity of the ubiquitous enzyme, tissue transglutaminase (TG2), is consistently seen in a variety of models of apoptosis. The p53 oncoprotein is also involved in apoptosis. Here we investigated the interaction of TG2 with p53 and show that the p53 is a substrate for the recently identified serine/threonine kinase activity of TG2. Phosphospecific antibodies indicated that TG2 phosphorylated p53 at Ser(15) and Ser(20), residues that are critically important in the interaction of p53 with Mdm2. The TG2-induced phosphorylation was abrogated by high Ca(2+) concentrations and inhibited by cystamine, a known inhibitor of TG2 cross-linking activity. Furthermore, we demonstrate that TG2-induced phosphorylation of p53 reduces the ability of p53 to interact with Mdm2. Although TG2 cross-linking activity has been clearly implicated in apoptosis, our observations reported here suggest TG2 modification of p53 could be an additional mechanism whereby TG2 could facilitate apoptosis. PMID- 16313887 TI - Metabolism of nilutamide in rat lung. AB - Nilutamide is a non-steroidal anti-androgen drug proposed in the treatment of metastatic prostatic carcinoma. Its therapeutic effects are overshadowed by the occurrence of adverse reactions, mediated by mechanisms that remain elusive. To elucidate possible mechanisms for nilutamide toxicity, we investigated the metabolism of nilutamide in rat lung homogenates, in subcellular fractions and in freshly isolated cells. In whole lung homogenates, the nitro group of nilutamide was reduced to the amine and hydroxylamine moieties. These conversions occurred exclusively in the absence of dioxygen, were increased by the addition of FMN, FAD, or NADPH. Reductive metabolism of nilutamide to the amine and hydroxylamine was further evidenced in subcellular fractions obtained by differential ultracentrifugation. It was found to take place mainly in the cytosol of rat lung and to be stimulated, strongly, upon co-addition of NADPH and FMN. Addition of inhibitors of enzymes involved in the reductive metabolism of nitroaromatic compounds indicated that reduction of nilutamide involved, mainly, soluble flavoproteins. Incubations with freshly isolated lung cells revealed that macrophages were the main players in nitroreduction of nilutamide whereas the epithelial type II cells and the non-ciliated Clara cells were less efficient in catalyzing this reaction. Our results show that nilutamide is extensively reduced by lung tissues in the absence of oxygen, especially by enzymes found in alveolar macrophages. In accordance with recent findings, subcellular localization, oxygen sensitivity, cofactor requirements and inhibitor studies lead us to suggest the involvement of a soluble nitric oxide synthase in lung cytosolic nitroreduction. PMID- 16313888 TI - Lorazepam and MK-801 effects on behavioral and electrographic indices of alcohol withdrawal sensitization. AB - Repeated cycles of chronic ethanol exposure and withdrawal result in sensitization of withdrawal-related CNS hyperexcitability that generally reflects an imbalance in activity of GABA and glutamate systems. Many pharmacological treatments for ethanol withdrawal target neuroadaptive changes in GABA and glutamate neurotransmission. The present study utilized a mouse model of repeated withdrawals to evaluate the ability of lorazepam and MK-801 treatments to antagonize behavioral and electroencephalographic (EEG) measures of sensitized withdrawal seizure activity. Adult male C3H/He mice received chronic intermittent ethanol vapor exposure in inhalation chambers (16 h/day) and during each withdrawal cycle, separate groups of mice were evaluated for handling-induced convulsions (HIC) or abnormal EEG (high-voltage "brief spindle episodes" (BSE)) activity. Lorazepam (0.5-1.0 mg/kg) or MK-801 (0.1-0.3 mg/kg) treatment at 1 h into each of three withdrawal cycles reduced behavioral (HIC) and electrographic (BSE) signs of seizure activity in a dose-related fashion compared to vehicle treated mice. During a subsequent untreated withdrawal, mice previously treated with lorazepam or MK-801 for earlier withdrawals exhibited reduced HIC activity during the acute phase but exacerbated HIC activity during the protracted phase of this final (fourth) withdrawal cycle. Both lorazepam and MK-801 treatment conditions resulted in enhanced BSE activity during the entire fourth (untreated) withdrawal episode. Collectively, these results suggest that while treatment of repeated ethanol withdrawals with a benzodiazepine (lorazepam) or an NMDA receptor antagonist (MK-801) may have some initial benefits in ameliorating the development of sensitized withdrawal excitability, such treatment may also render subjects more vulnerable to seizure activity at later time points. PMID- 16313889 TI - Do bone marrow stromal cells proliferate after transplantation into mice cerebral infarct?--a double labeling study. AB - The present study was aimed to clarify the proliferation capacity of the bone marrow stromal cells (BMSC) transplanted into the brain. The BMSC were harvested from green fluorescence protein (GFP)-transgenic mice, grown to the confluency and passed three times. They were labeled by co-culture with Ferucarbotran, a superparamagnetic iron oxide (SPIO) agent. The proportions of the SPIO-positive cells were evaluated from P3 to P7, using Turnbull blue staining. The GFP-BMSC labeled by Ferucarbotran were transplanted into the ipsilateral striatum of the mice brain subjected to permanent focal ischemia at 7 days after the insult. The distribution and differentiation of GFP- and SPIO-positive cells in the brain were studied 3 months after transplantation, using immunohistochemistry and Turnbull blue staining. As the results, the proportions of the SPIO-positive cells gradually decreased from 93.6% at P3 to 6.5% at P7. Fluorescence immunohistochemistry revealed that the GFP-positive cells were widely distributed around infarct and partially expressed MAP2 and NeuN 3 months after transplantation. However, only a smaller number of SPIO-positive cells could be detected on Turnbull blue staining. The ratio of the SPIO- to GFP-positive cells was approximately 2.7%. The results strongly suggest that the BMSC repeat proliferation many times, migrate into the lesion, and partially express the neuronal phenotype in the host brain during 3 months after transplantation. The double labeling technique would be valuable to prove the proliferation of the transplanted cells in the host tissue because GFP gene and SPIO nanoparticles have different inheritance characteristics. PMID- 16313891 TI - Structures of two putative O-specific polysaccharides from the Rahnella aquatilis 3-95 lipopolysaccharide. AB - Two polysaccharide preparations (OPSI and OPSII) were obtained by mild acid degradation of the lipopolysaccharide of Rahnella aquatilis 3-95. Studies by chemical methods and 1H and 13C NMR spectroscopy showed that OPSI is a linear alpha-D-mannan having a trisaccharide repeat and OPSII is a approximately 2:1 mixture of the same mannan and an alpha-d-glucan: PMID- 16313892 TI - Comparative study of alkali- and acidic organic solvent-soluble hemicellulosic polysaccharides from sugarcane bagasse. AB - Two-stage treatments of sugarcane bagasse with mild alkali and acidic 1,4-dioxane were performed. Pretreatment with 1M NaOH aqueous solution at 20, 25, 30, 35, and 40 degrees C for 18 h released 55.5%, 57.3%, 59.1%, 60.9%, and 62.1% of the original hemicelluloses, respectively. Post-treatment of the corresponding alkali treated residue with 1,4-dioxane-2M HCl (9:1, v/v) at 87 degrees C for 2h, respectively, degraded 11.6%, 11.9%, 11.4%, 10.9%, and 10.6% of hemicelluloses (% dry starting material). It was found that the five alkali-soluble hemicellulosic preparations contained a much higher amounts of xylose (78.0-82.2%) and slightly higher uronic acids (4.8-5.8%), mainly 4-O-methyl-alpha-d-glucopyranosyluronic acid, but were lower in arabinose (9.3-11.7%) and glucose (2.2-4.1%) than those of the corresponding five acidic dioxane-degraded hemicellulosic fractions in which xylose (44.9-46.8%), arabinose (35.9-38.1%), and glucose (13.0-13.7%) were the major sugar constituents. The studies revealed that the five alkali-soluble hemicellulosic preparations were more linear and acidic, and had a large molecular weight (35,200-37,430 g mol(-1)) than those of the hemicellulosic fractions (12,080-13,320 g mol(-1)) degraded during the acidic dioxane post treatment. This demonstrated that the post-treatment with acidic dioxane under the condition used resulted in substantial degradation of the hemicellulosic polymers. The 10 hemicellulosic samples were further characterized by FT-IR and 1H and 13C NMR spectroscopy, GPC and thermal analysis, and the results are reported. PMID- 16313893 TI - Two different O-polysaccharides from Escherichia coli O86 are produced by different polymerization of the same O-repeating unit. AB - The structure of a new O-polysaccharide from Escherichia coli O86:K62:B7 was determined using NMR and methylation analysis. The structure is as follows: [carbohydrate: see text]. Comparison with the previously published structure from E. coli O86:K2:H2 revealed that the O-polysaccharides from these two E. coli O86 serotypes share the same branched pentasaccharide repeating unit. However, they differ in the anomeric configuration of the linkage, the linkage position, and the identity of the residue through which polymerization occurs. The immunochemical activity of these two forms of LPS toward anti-B antibody was studied and compared. The results showed that LPS from E. coli O86:K2:H2 strain possesses higher blood group B reactivity. The immunoreactivity difference was explained by modeling of the O-repeating unit tetrasaccharide fragments. This finding provides a good system for the further study of O-polysaccharide biosynthesis especially the repeating unit polymerization mechanism. PMID- 16313894 TI - Relationships between circulating matrix metalloproteinase-2 and -9 and renal function in patients with chronic kidney disease. AB - BACKGROUND: It has been proven that extracellular matrix turnover is involved in the pathogenesis of various renal fibrosis diseases. Matrix metalloproteinase-2 and -9 (MMP-2 and -9) are the extracellular matrix degrading enzymes that are believed to play important roles in renal diseases. However, the relationship of circulating levels of MMP-2, -9 and serum creatinine in the patients of chronic kidney disease (CKD) has not yet been investigated. METHODS: Gelatin zymography and ELISA were employed to measure MMP-2 and MMP-9 activities in the plasma samples of 60 CKD patients and 40 control subjects. RESULTS: Serum creatinine concentrations and MMP-2 activities were significantly higher (p<0.001) while MMP 9 activity and creatinine clearance (CCr) were significantly lower (p<0.05 and p<0.001, respectively) in CKD patients, as compared with those of control subjects. In addition, serum creatinine concentrations correlated with MMP-2 activity (R=0.288, p<0.05) and inversely correlated with that of MMP-9 (R=0.344, p<0.01). CONCLUSIONS: This study demonstrated a correlation between MMP-2, -9 and serum creatinine in CKD patients to suggest that MMP-2 and MMP-9 might contribute in the pathogenesis of CKD. PMID- 16313895 TI - Markers of bone metabolism in congestive heart failure. AB - Congestive heart failure (CHF) is a chronic disease, whose incidence is especially growing in the subpopulation of elderly people. CHF is characterized by dyspnea and fatigue at rest or with exertion, ankle swelling and pulmonary edema. Cardiac transplantation is the ultimate therapeutic measure in patients with end-stage CHF. Some risk factors associated with CHF such as low mobility, renal failure, and prescription of specific drugs may predispose patients to develop osteoporosis. This review article gives an overview about markers of bone metabolism in CHF patients as well as in heart transplant recipients. At first, the physiology of bone metabolism is summarized. Then, a short description of different bone formation and resorption markers is presented. They can be used to characterize actual bone metabolism and can be helpful to explain possible mechanisms of bone loss. Regarding pre-transplant CHF patients, available data indicate that the disturbances in bone metabolism are only subtle. Heart transplant recipients, however, are at increased risk for osteoporotic bone loss due to the use of immunosuppressive agents such as corticosteroids and calcineurin inhibitors. Preventive strategies are able to normalize bone metabolism and to attenuate the high bone loss during the first year after heart transplantation. PMID- 16313896 TI - Fertilization regulates apoptosis of Ciona intestinalis extra-embryonic cells through thyroxine (T4)-dependent NF-kappaB pathway activation during early embryonic development. AB - In Ciona intestinalis, the elimination of extra-embryonic test cells during early stage of development is delayed by a fertilization signal. Test cells undergo a caspase-dependent apoptosis event repressed by thyroxine (T4)-activated NF kappaB. When apoptosis was experimentally blocked, the hatching stage was delayed. The incubation of unfertilized eggs with a 1-h-fertilized egg extract or purified T4 restored apoptosis in test cells at a similar timing than found in fertilized eggs. Ciona expresses specific genes forming a functional IkappaB/NF kappaB pathway. One, Ci-p65, was transiently induced upon fertilization via T4 and found to exert its anti-apoptotic role in test cells nuclei as well as in a reconstituted cell system. Blocking NF-kappaB activity by dexamethasone-induced overexpression of Ci-IkappaB abrogated the repression of apoptosis in test cells. Overall, the data are consistent for defining a central coupling role of both T4 and NF-kappaB during early embryo development. PMID- 16313897 TI - The Drosophila Par domain protein I gene, Pdp1, is a regulator of larval growth, mitosis and endoreplication. AB - PDP1 is a basic leucine zipper (bZip) transcription factor that is expressed at high levels in the muscle, epidermis, gut and fat body of the developing Drosophila embryo. We have identified three mutant alleles of Pdp1, each having a similar phenotype. Here, we describe in detail the Pdp1 mutant allele, Pdp1(p205), which is null for both Pdp1 RNA and protein. Interestingly, homozygous Pdp1(p205) embryos develop normally, hatch and become viable larvae. Analyses of Pdp1 null mutant embryos reveal that the overall muscle pattern is normal as is the patterning of the gut and fat body. Pdp1(p205) larvae also appear to have normal muscle and gut function and respond to ecdysone. These larvae, however, are severely growth delayed and arrested. Furthermore, although Pdp1 null larvae live a normal life span, they do not form pupae and thus do not give rise to eclosed flies. The stunted growth of Pdp1(p205) larvae is accompanied by defects in mitosis and endoreplication similar to that associated with nutritional deprivation. The cellular defects resulting from the Pdp1(p205) mutation are not cell autonomous. Moreover, PDP1 expression is sensitive to nutritional conditions, suggesting a link between nutrition, PDP1 isotype expression and growth. These results indicate that Pdp1 has a critical role in coordinating growth and DNA replication. PMID- 16313898 TI - The C. elegans histone deacetylase HDA-1 is required for cell migration and axon pathfinding. AB - Histone proteins play integral roles in chromatin structure and function. Histones are subject to several types of posttranslational modifications, including acetylation, which can produce transcriptional activation. The converse, histone deacetylation, is mediated by histone deacetylases (HDACs) and often is associated with transcriptional silencing. We identified a new mutation, cw2, in the Caenorhabditis elegans hda-1 gene, which encodes a histone deacetylase. Previous studies showed that a mutation in hda-1, e1795, or reduction of hda-1 RNA by RNAi causes defective vulval and gonadal development leading to sterility. The hda-1(cw2) mutation causes defective vulval development and reduced fertility, like hda-1(e1795), albeit with reduced severity. Unlike the previously reported hda-1 mutation, hda-1(cw2) mutants are viable as homozygotes, although many die as embryos or larvae, and are severely uncoordinated. Strikingly, in hda-1(cw2) mutants, axon pathfinding is defective; specific axons often appear to wander randomly or migrate in the wrong direction. In addition, the long range migrations of three neuron types and fasciculation of the ventral nerve cord are defective. Together, our studies define a new role for HDA-1 in nervous system development, and provide the first evidence for HDAC function in regulating neuronal axon guidance. PMID- 16313899 TI - Up-regulation of nicotinic acetylcholine receptors by central-type acetylcholinesterase inhibitors in rat cortical neurons. AB - We previously reported that donepezil, a central-type acetylcholinesterase inhibitor, showed neuroprotective action via alpha4-and alpha7-nicotinic acetylcholine receptors against glutamate neurotoxicity in rat cortical culture. The present study was performed to investigate whether the neuroprotective action of acetylcholinesterase inhibitors is accompanied by the alteration of expression and function of nicotinic receptors. Four days treatment with acetylcholinesterase inhibitors (10 microM) enhanced the nicotine-induced increase of the intracellular calcium concentration. Immunoblot analysis revealed that donepezil increased both alpha4 and alpha7 subunit proteins. Donepezil and galanthamine increased the number of cells expressing alpha4- and alpha7 nicotinic receptors in immunocytochemical analysis. We examined whether up regulation of nicotinic receptors affected the neuroprotective action of acetylcholinesterase inhibitors. Under up-regulating conditions, donepezil and galanthamine exerted neuroprotective action at lower concentrations. These results suggest that donepezil and galanthamine up-regulate nicotinic receptors in cortical neurons, and that the up-regulation of nicotinic receptors may make cortical neurons more sensitive to the neuroprotective action of donepezil and galanthamine. PMID- 16313900 TI - Stimulation of 5-HT 1A receptors increases the seizure threshold for picrotoxin in mice. AB - To evaluate the possible role of 5-HT 1A and 5-HT 2A receptors in the anticonvulsant effect of swim stress, mice were pre-treated with agonists and antagonists of these receptors prior to exposure to stress and the intravenous infusion of picrotoxin. 8-OH-DPAT ((+/-)-8-hydroxy-2-(di-n-propylamino) tetralin) and WAY-100635 (a selective agonist and antagonist of 5-HT 1A receptors), DOI (1 (2,5-dimethoxy-4-iodophenyl)-2-aminopropane) and ketanserin (a 5-HT 2A/2C receptor agonist and antagonist) were used. Results demonstrated that 1 and 3 mg/kg of 8-OH-DPAT increased the doses of picrotoxin producing running/bouncing clonus, tonic hindlimb extension and death in stressed and unstressed mice, respectively. Pre-treatment with WAY (0.3 mg/kg) prevented the effect of 8-OH DPAT (3 mg/kg). DOI (2.5 mg/kg) and ketanserin (1 mg/kg) failed to affect the seizure threshold for picrotoxin. The results show that stimulation of 5-HT 1A receptors exerts anticonvulsant actions in stressed and unstressed mice, while stimulation of 5-HT 2A/2C receptors does not interfere with the effect of stress on picrotoxin-induced convulsions. PMID- 16313901 TI - Endothelial antioxidant actions of dihydropyridines and angiotensin converting enzyme inhibitors. AB - Dihydropyridines and angiotensin converting enzyme inhibitor effects on superoxide and nitric oxide (NO) were compared in high glucose (20 mM, 24 h) treated human Ea.hy 926 endothelial cells. High glucose stimulated superoxide both extracellularly (lucigenin chemiluminescence, cytochrome c reduction) and intracellularly (dihydrorhodamine 123 fluorescence). The dihydropyridines amlodipine, nisoldipine, BayK 8644 or the angiotensin converting enzyme inhibitors captopril and enalaprilat attenuated extra- and intracellular superoxide formation; nifedipine blocked extracellular increases only, ramiprilat was without antioxidant effect. Dihydropyridines and captopril also prevented NADPH-driven superoxide release. Antioxidant actions were blunted by a bradykinin B(2) receptor antagonist or an inhibitor of p38 mitogen activated protein kinase (MAPK), and were accompanied by improved NO release (amperometric sensor). p38MAPK inhibition prevented the NO-sparing actions of dihydropyridines but not angiotensin converting enzyme inhibitors. Thus, dihydropyridines and angiotensin converting enzyme inhibitors limit high glucose-induced superoxide formation and improve NO bioavailability in human endothelial cells, in part via bradykinin and p38MAPK. PMID- 16313902 TI - Characterisation of alpha2-adrenoceptor subtypes involved in gastric emptying, gastric motility and gastric mucosal defence. AB - The effect of clonidine on ethanol-induced gastric mucosal damage, gastric emptying and gastric motility was compared. The clonidine-induced gastroprotective effect (0.03-0.09 micromol/kg, s.c.) was antagonised by yohimbine (5 micromol/kg, s.c.), prazosin (0.23 micromol/kg; alpha2B-adrenoceptor antagonist) and naloxone (1.3 micromol/kg, s.c.). Clonidine also inhibited the gastric emptying of liquid meal (0.75-3.75 micromol/kg, s.c.) and gastric motor activity (0.75 micromol/kg, i.v.) stimulated by 2-deoxy-D-glucose (300 mg/kg, i.v.). Inhibition of gastric emptying and motility was reversed by yohimbine (5 and 10 micromol/kg, s.c., respectively), but not by prazosin (0.23 micromol/kg, s.c.) or naloxone (1.3 micromol/kg, s.c.). Oxymetazoline-an alpha2A-adrenoceptor agonist-inhibited both gastric emptying (0.67-6.8 micromol/kg, s.c.) and motility (0.185-3.4 micromol/kg, i.v.), whereas it failed to affect gastric mucosal lesions. The results indicate that in contrast to the gastroprotective effect, which is mediated by alpha2B-adrenoceptor subtype, alpha2A-adrenoceptor subtype may be responsible for inhibition of gastric emptying and motility. However, the site of action (central, peripheral, both) remains to be established. PMID- 16313903 TI - 2-(2-Br-phenyl)-8-methoxy-benzoxazinone (HPW-RX2), a direct thrombin inhibitor with a suppressive effect on thromboxane formation in platelets. AB - 2-(2-Br-phenyl)-8-methoxy-benzoxazinone (HPW-RX2), a newly synthetic benzoxazinone derivative, has previously been shown to inhibit rabbit platelet aggregation caused by thrombin and arachidonic acid. In the present study, the mechanism for the antiplatelet effect of HPW-RX2 was further investigated. In human platelets, HPW-RX2 concentration-dependently inhibited platelet aggregation, ATP release, P-selectin expression, and intracellular calcium mobilization caused by thrombin. In contrast, HPW-RX2 had no significant effect on either SFLLRN- or GYPGKF-induced platelet aggregation, indicating that HPW-RX2 did not interfere with platelet thrombin receptors. Moreover, HPW-RX2 inhibited the amidolytic activity of thrombin and prolonged the fibrinogen clotting time. These results suggest that the inhibitory effect of HPW-RX2 on thrombin-induced platelet aggregation is via direct inhibition of thrombin proteolytic activity. Besides the inhibition on thrombin, HPW-RX2 also prevented platelet aggregation, ATP release, and increase in [Ca2+]i caused by arachidonic acid and low concentration collagen. In a parallel manner, both arachidonic acid-induced thromboxane B2 and prostaglandin D2 formations were decreased in platelets treated with HPW-RX2. This indicates that HPW-RX2 is able to inhibit the arachidonic acid cascade at the cyclooxygenase level. This is the first report of a benzoxazinone derivative possessing both thrombin and cyclooxygenase inhibitory properties. The dual effect of HPW-RX2 might provide extra therapeutic benefits for treatment of arterial thrombosis. PMID- 16313904 TI - Leishmania amazonensis: chemotaxic and osmotaxic responses in promastigotes and their probable role in development in the phlebotomine gut. AB - Taxic responses may play a role in development of Leishmania in their phlebotomine sand fly vectors. They are possibly responsible for movement of the parasites towards the anterior regions of the gut, from where they would be transmitted to the vertebrate host. A methodology capable to distinguish chemotaxic from osmotaxic responses was described and used to characterise taxic responses in Leishmania promastigotes. These were able to respond to chemotaxic as well as to osmotaxic stimuli. Like bacteria, promastigotes were capable to undergo "adaptation," a phenomenon by which they stop responding to a continuos stimulus. A model capable to explain how a relatively small number of different receptors works to perceive gradients in chemotaxic responses was proposed. According to this model, these receptors possess low specificity and a wide range of affinities varying from high to low. A low specificity makes the same receptor able to bind to a large number of different but structurally related molecules and; a wide range of affinities (considering a population of receptors), implies that the number of receptors "occupied" by attractant molecules along a gradient would go growing step by step. PMID- 16313905 TI - Modulation of p53 expression and its role in the conversion to a fully immortalized chicken embryo fibroblast line. AB - We have established a spontaneously immortalized chicken embryo fibroblast (CEF) cell line (SC-1) that has been in continuous culture for more than three years. This is only the second report of a spontaneously immortalized reverse transcriptase (RT)-negative chicken cell line. The SC-1 cells emerged from crisis (at about passage 29-31) with a slower growth rate than primary cells. Passage 50 SC-1 cells expressed similar levels of p53 mRNA, but slightly lower levels of p53 protein than passage 6 CEF cells. By passage 120, p53 mRNA levels were significantly decreased in the SC-1 cells, while protein levels were slightly increased compared to passage 6 CEF cells. However, functional analysis of p53 revealed reduced activity in later passage SC-1 cells. Other p53-related genes including p21WAF1, p27Kip1, MDM-2, and the p16INK4a alternate reading frame (ARF) sequence showed similar patterns of differential mRNA expression. Levels of p15INK4b mRNA and protein were dramatically decreased in SC-1 cells, suggesting that the Rb pathway also has been compromised. Telomerase expression was undetectable in SC-1 cells. Fluorescence-activated cell sorting analysis showed that SC-1 and primary cells contained a similar proportion of G0/G1 phase cells, unlike the only other spontaneously immortalized chicken cell line (DF-1). The present study suggests that alterations in the p53 and Rb pathways cause fluctuations in expression levels of important cell-cycle regulatory genes during crucial transition periods as the SC-1 spontaneously immortalized chicken fibroblast cells progress toward becoming a fully committed cell line. PMID- 16313906 TI - Valproic acid-mediated Hsp70 induction and anti-apoptotic neuroprotection in SH SY5Y cells. AB - Valproic acid (VPA), an anticonvulsant and mood-stabilizing drug, has been reported to exert neuroprotection against a variety of insults. We now show that VPA attenuates rotenone (a potent complex I inhibitor)-induced apoptosis through the induction of heat shock protein 70, which may interact with apoptotic protease-activating factor 1. Activation of p-Akt, p-Bcl-2, as well as p-Erk1/2 by VPA may be co-contributors to the protection. PMID- 16313907 TI - Elevated testosterone levels and social ranks in female rock hyrax. AB - In mammals, males maintain significantly higher testosterone (T, 'the male hormone') levels than females throughout the year and are typically dominant over females. Our study of the cooperatively breeding rock hyrax (Procavia capensis) demonstrated a distinct exception. In most hyrax social groups that we studied, adult females were at the highest social rank within the group and showed higher than or equal T levels to males. To our knowledge, this is the first reported instance of adult female mammals demonstrating higher T levels than adult males. However, although T levels significantly correlated with rank in males, in females such correlations were not detected, suggesting a more complex interplay between behavior and endocrine factors in this species. PMID- 16313908 TI - Treatment of obesity with diet/exercise versus pioglitazone has distinct effects on lipoprotein particle size. AB - OBJECTIVE: We compared the effects of diet/exercise induce weight loss versus pioglitazone on lipoprotein particle characteristics in non-diabetic, insulin resistant adults to determine whether the responses and potential mechanisms for improvement are similar between these two insulin sensitizing regimens. METHODS AND RESULTS: Thirty-seven obese, insulin resistant volunteers were randomized to 19 weeks of either pioglitazone or diet/exercise treatment. Insulin sensitivity (Si) by IVGTT and lipoprotein subclass particle profiles by NMR was measured. Only diet/exercise lowered TG (-40+/-11 mg/dL), total (-29+/-4 mg/dL) and LDL cholesterol (-20+/-4 mg/dL; all p<0.01). Both diet/exercise and pioglitazone improved Si (p<0.05), decreased total LDL particles (-227+/-40 and -149+/-21 nmol/L, respectively, both p<0.001) and small LDL particles (-242+/-58 and -226+/ 36, respectively, both p<0.001). Large LDL particle concentrations increased with both treatments. LDL particle subclasses changes correlated with changes in Si only in the diet/exercise group. CONCLUSION: In obese, insulin resistant non diabetic adults, pioglitazone improved LDL particle characteristics in a manner similar to diet/exercise without changing TG, diabetic control, total or LDL cholesterol. This effect could have implications for atherosclerotic risk. PMID- 16313909 TI - Platelet activation is associated with hypoadiponectinemia and carotid atherosclerosis. AB - Adiponectin, an adipokine secreted specifically from adipose tissue, has plurifunctions including antidiabetic, antiatherosclerotic, and antiinflammatory functions. Recently, platelet activation and the subsequent local inflammation have been implicated in progression of atherosclerosis. The aim of the study is to examine the interrelation among plasma adiponectin levels, platelet activation status and quantitatively determined carotid atherosclerosis. Subjects (n = 277) including 136 type 2 diabetic, 138 hypertensive, and 203 hypercholesterolemic patients participated in the study. Platelet activation was determined as percentage of polymorphonuclear cells (PMNs) or monocytes aggregated with platelets analyzed by CD41-positivity determined by whole-blood flow cytometry. PMN-platelet aggregates were significantly and positively associated with carotid atherosclerosis (intimal-medial thickness, IMT) with the interaction stronger than that of monocyte-platelet aggregates. Stepwise regression analyses revealed that PMN-platelet aggregates were the third strongest determinant of carotid IMT, with age and HbA1c stronger independent determinants. Simple and stepwise regression analyses of the factors associated with PMN-platelet aggregates revealed that HbA1c (r = 0.423), serum adiponectin levels (r = -0.289) and age (r = -0.184) were the three independent determinants. Thus, our data unveil novel link between hypoadiponectinemia and platelet activation. PMID- 16313910 TI - Cardiovascular disease in the polycystic ovary syndrome: new insights and perspectives. AB - The new millennium has brought intense focus of interest on the risk of cardiovascular disease in women. The polycystic ovary syndrome (PCOS) is a common endocrine disorder in women characterised by hyperandrogenism and oligomenorrhoea. Most women with PCOS also exhibit features of the metabolic syndrome, including insulin resistance, obesity and dyslipidaemia. While the association with type 2 diabetes is well established, whether the incidence of cardiovascular disease is increased in women with PCOS remains unclear. Echocardiography, imaging of coronary and carotid arteries, and assessments of both endothelial function and arterial stiffness have recently been employed to address this question. These studies have collectively demonstrated both structural and functional abnormalities of the cardiovascular system in PCOS. These alterations, however, appear to be related to the presence of individual cardiovascular risk factors, particularly insulin resistance, rather than to the presence of PCOS and hyperandrogenaemia per se. However, given the inferential nature of the evidence to date, more rigorous cohort studies of long-term cardiovascular outcomes and clinical trials of risk factor modification are required in women with PCOS. PMID- 16313911 TI - Lipid metabolism mediated by adipocyte lipid binding protein (ALBP/aP2) gene expression in human THP-1 macrophages. AB - The critical initiating event in atherogenesis involves the invasion of monocytes through the endothelial wall of arteries, and their transformation from macrophages into foam cells. Human THP-1 monocytic cells can be induced to differentiate into macrophages by phorbol myristate acetate (PMA) treatment, and can then be converted into foam cells by exposure to oxidized low-density lipoprotein (oxLDL). We previously reported that adipocyte lipid binding protein (ALBP/aP2) is a gene that is highly up-regulated in foam cells in response to oxLDL. Here, we showed that overexpression of the ALBP gene using a lentiviral construct in macrophage foam cells enhanced the accumulations of cholesterol and triglyceride, probably due to an increased expression of the scavenger receptor type AI (SR-AI), which plays an important role in cell lipid metabolism. Moreover, we determined that the expression of acyl-coenzyme A: cholesterol acyltransferase 1 (ACAT1) gene was up-regulated by the overexpression of ALBP gene, and on the other hand, the ATP-binding cassette A1 (ABCA1) gene and hormone sensitive lipase (HSL) gene, which mediate separately cholesterol efflux and cholesterol ester hydrolysis in the macrophage cells, were down-regulated by the overexpression of ALBP gene in these cells. Finally, our data indicated that oxLDL regulates expression of ALBP related to two peroxisome proliferator responsive elements (PPREs) which are located in ALBP promoter region. These results have determined that ALBP gene expression accelerates cholesterol and triglyceride accumulation in macrophage foam cells and affects some key gene expression for lipid metabolism, suggesting some pivotal roles of ALBP in lipid metabolism for macrophage foam cell formation. PMID- 16313912 TI - Dietary antioxidants in preventing atherogenesis. AB - Several naturally occurring constituents have received considerable attention because of their potential antioxidant activity. Consuming a diet rich in natural antioxidants has been associated with prevention from and/or treatment of atherosclerosis. Bioactive components of food, which are of special interest, include the Vitamins E and C, polyphenols, carotenoids-mainly lycopene and beta carotene, and coenzyme Q10, featured by antioxidant properties. Antioxidant therapy is supposed to be effective in the early stages of atherosclerosis by preventing LDL oxidation and the oxidative lesion of endothelium. This review focuses on the effect of dietary antioxidants pertained to LDL oxidation and to the vascular endothelial dysfunction. Now that the human genome has been completely sequenced, genetic factors involved in oxidation may open new horizons to identify persons at risk for cardiovascular disease, allowing effective dietary intervention strategies to recover normal homeostasis and to prevent diet related implications. On this basis, current studies on the action of selected antioxidant nutraceuticals on the activity of transcription factors, such as final targets in the signal transduction cascade and gene regulation, may emerge into new treatment concepts. PMID- 16313913 TI - Influence of glenohumeral mismatch on bone strains and implant displacements in implanted glenoids. An in vitro experimental study on cadaveric scapulae. AB - In shoulder arthroplasty, there is no consensus about the ideal mismatch between a prosthetic humeral head and a glenoid component. Thus, investigations into mismatch effects from a biomechanical point of view can be useful. The aim of this in vitro study was to help us understand mismatch influence on bone strains, translational forces in the joint and implant/bone displacements in implanted scapulae. Five fresh cadaveric scapulae were implanted with a cemented keeled polyethylene implant. The lower part of the scapulae was embedded and the loadings were carried out using five metallic spheres simulating mismatches of 0, 2, 4, 5 and 6 mm. Loadings included a constant compressive preload of 392N and an anterior, posterior, inferior and superior translation of 2.5 mm. We measured the transversal force necessary to produce the imposed translation, the strains at six locations around the peripheral cortex of the glenoid using strain gages and the relative implant/bone displacements using CCD cameras. Generally, the increase of mismatch reduced the translational forces, the strains around the glenoid and, except for the anterior loading, the relative implant/bone displacements. Few and even no significant differences were observed when the mismatch varied from 0 to 2 mm; the number of significant differences increased when the mismatch varied from 0 to 4mm and from 0 to 5 mm; the results obtained for a 0-6 mm variation in mismatch were comparable to those obtained for a 0-5 mm variation. This study underlines that the mismatch has a significant effect on bone strains, relative implant/bone displacements and induced translational forces when a compressive preload and imposed translations were applied on implanted scapulae. PMID- 16313914 TI - A validated model of passive muscle in compression. AB - A better characterisation of soft tissues is required to improve the accuracy of human body models used, amongst other applications, for virtual crash modelling. This paper presents a theoretical model and the results of an experimental procedure to characterise the quasi-static, compressive behaviour of skeletal muscle in three dimensions. Uniaxial, unconstrained compression experiments have been conducted on aged and fresh animal muscle samples oriented at various angles from the fibre direction. A transversely isotropic hyperelastic model and a model using the theory of transverse isotropy and strain dependent Young's moduli (SYM) have been fitted to the experimental data. Results show that the hyperelastic model does not adequately fit the data in all directions of testing. In contrast, the SYM gives a good fit to the experimental data in both the fibre and cross fibre direction, up to 30% strain for aged samples. The model also yields good prediction of muscle behaviour at 45 degrees from the fibre direction. Fresh samples show a different behaviour than aged tissues at 45 degrees from the fibre direction. However, the SYM is able to capture this difference and gives a good fit to the experimental data in the fibre, the cross-fibre and at 45 degrees from the fibre direction. The model also yields good prediction of muscle behaviour when compressed at 30 degrees and 60 degrees from the fibre direction. The effect of the time of test after death has also been investigated. Significant stiffening of muscle behaviour is noted a few hours after death of the subject. PMID- 16313915 TI - Apple (Malus pumila) procyanidins fractionated according to the degree of polymerization using normal-phase chromatography and characterized by HPLC-ESI/MS and MALDI-TOF/MS. AB - Our previously reported method for the fractionation of apple procyanidins was modified successfully to achieve the separation of (epi)catechins and procyanidins (ranging from dimers to octamers) according to the degree of polymerization. Normal-phase chromatography was employed, using a hexane-methanol ethyl acetate mixture as the mobile phase. Each fraction was characterized using high-performance liquid-chromatography electrospray-ionization mass spectrometry (HPLC-ESI/MS) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS). This method will be useful for the evaluation of the physiological functions of proanthocyanidins and for the elucidation of their individual structures. PMID- 16313916 TI - Analysis of mass transport models for protein adsorption to cation exchanger by visualization with confocal laser scanning microscopy. AB - The mass transfer of bovine serum albumin (BSA) to a cation exchanger, SP Sepharose FF, has been studied by finite batch adsorption experiments. The uptake curve was simulated with three mass transport models (i.e., effective pore diffusion model, surface diffusion model and Maxwell-Stefan model) incorporating the particle size distribution of the adsorbent particles. All the three models can simulate the uptake curves reasonably well. However, how well these models could simulate the real concentration profile within the adsorbent particle cannot be verified by the fitness of the models to the uptake curve. Thus, confocal laser scanning microscopy (CLSM) was used to visualize protein uptake to the porous adsorbent particles during the batch experiments. Using a fluorescent dye-labeled bovine serum albumin (BSA) for the dynamic adsorption experiments, the radial concentration profiles of the labeled BSA molecules into individual adsorbent particles at different times were obtained from the CLSM images. The protein distribution profiles within various particle diameters at different time were compared with the radial protein distributions predicted from the models. It reveals that surface diffusion model describes the intraparticle protein concentration profiles better than the other two models. PMID- 16313918 TI - The effect of inorganic particles on slot die coating of poly(vinyl alcohol) solutions. AB - Among various coating processes, slot die coating belongs to a class of pre metered coating, in which the coating film thickness can be predetermined. In the past, most research works on slot die coating have focused mainly on polymer solutions; very little information is available using suspensions as coating fluids. In this study, the effect of adding TiO2 and SiO2 in aqueous polyvinyl alcohol (PVA) solutions on slot die coating is investigated. It was found that the stable coating window was enlarged with the addition of particles, and its size increased with solids concentration. This is due to the strong interaction between polymer and particles, resulting in a higher viscosity and surface tension. As a result, the upstream coating bead is more stable and the maximum coating speed is extended to a high value, hence the coating window becomes larger. Although both viscosity and surface tension appear to contribute to the stability of coating flow, the effect of surface tension is more dominant. The surface tension of a suspension with porous particles was higher than one with hard solid particles. Consequently, the coating window obtained with the former was significantly larger than the latter. Flow visualization revealed that under the same operating conditions, the upstream dynamic contact angle for the suspension was smaller than for the aqueous polymer solution. This observation could be related to the stability of the upstream coating bead, and hence the coating window. The experimental flow fields were verified numerically with the aid of a numerical simulation package (Flow-3D). PMID- 16313919 TI - Electron transfer behavior and water photodecomposition ability of calcined material from a cerium-S-phenylene-O-holmium-O-phenylene-S hybrid copolymer. AB - Calcination of a cerium-S-phenylene-O-holmium-O-phenylene-S hybrid copolymer under a vacuum gave cerium oxide-carbon cluster-holmium oxide composite material. The material calcined at 600 degrees C loaded with Pt particles could decompose water to H2 and O2 with a H2/O2 ratio of 2 under visible light irradiation. ESR spectral examinations of the calcined materials revealed the possibility of a two step electron transfer in the process of CeO2 --> carbon cluster --> Ho2O3 --> Pt with an oxidation site at CeO2 particles and a reduction site at Pt particles. PMID- 16313920 TI - Synthesis of single-crystal beta-Ni(OH)2 nanodisks and alpha-Fe2O3 nanocrystals in C2H5OH-NaOH-NH3 x H2O system. AB - Circular beta-Ni(OH)2 nanodisks and rhombohedral and hexagonal alpha-Fe2O3 nanocrystals were prepared using the C2H5OH-NaOH-NH3 x H2O system under hydrothermal conditions. The C2H5OH/H2O solvent is an appropriate one for the growth of these two materials with their thermodynamically favored morphologies. The possible formation mechanisms are discussed. PMID- 16313921 TI - Preparation of highly monodisperse poly(methyl methacrylate) particles incorporating fluorescent rhodamine 6G for colloidal crystals. AB - Soap-free emulsion polymerization was extended to preparation of monodisperse poly(methyl methacrylate) (PMMA) particles incorporating rhodamine 6G (R6G) fluorescent molecules. The polymerization was conducted in the presence of an anionic monomer, p-styrenesulfonate (NaSS), which improved dispersion stability of the polymer particles. NaSS concentrations was ranged up to 2 mol/m3 H2O in the polymerization at 0.5 kmol/m3 H2O methyl methacrylate (MMA) monomer and 5 mol/m3 H2O potassium persulfate (KPS) initiator for R6G concentrations from 0.1 to 10 mol/m3-polymer. At R6G concentrations lower than 1.0 mol/m3-polymer, PMMA particles were highly monodisperse and incorporated most R6G molecules. The average sizes of PMMA particles were in a rage of 160-300 nm, and decreased with the concentration of NaSS. The high monodispersity of the particles enabled the fabrication of colloidal crystals of the particles with a vertical deposition method. PMID- 16313922 TI - Electrical penetration graphs of thrips revised: combining DC- and AC-EPG signals. AB - Within thrips feeding behaviour, sequences of four waveforms have been distinguished earlier in the DC-EPG, i.e. P, Q, R and S, representing mandibular stylet insertion, maxillary stylet insertion, ingestion, and repetitive mandibular insertion, respectively. During signal analysis it appeared that transitions from one waveform to the next were difficult to establish, making results ambiguous. In order to improve the quantitative reliability of the thrips' EPG data, the DC-EPGs were recorded concurrently with AC-EPG signals, thus providing two signals from the same activities containing different information. The additional AC information did not solve most quantification problems, however. We now propose to merge waveforms P, Q, and S, into 'puncture phase' (indicated by PQ) and waveforms R, T, and U, into 'feeding phase' (indicated by R), rather than trying to analyse all separate waveforms. This will provide a more reliable and much less laborious analysis of thrips probing behaviour. Waveforms T and U are two novel waveforms identified here by combining DC- and AC-EPG recordings with concurrent video recordings. Waveform T represents a single mandibular thrust embedded in waveform R and waveform U represents the end of a probe, presumably the retraction of the maxillary stylets. PMID- 16313923 TI - Anxiety- and novelty seeking-related personality traits and serotonin transporter gene polymorphisms. AB - The affection of human personality by the promoter and the intron 2 polymorphism in the serotonin transporter gene (SERT) is inconsistently reported. We aimed to clarify this situation by gender-specific haplotype-phenotype association. 98 women and 97 men completed the personality inventories NEO-PI-R and TPQ. The subjects were genotyped for the two SERT polymorphisms and the haplotypes were calculated. The short (S) and long (L) promoter alleles and the 12 and 10 repeat intron 2 alleles formed the haplotypes S 12, S 10, L 12 and L 10. In men, scores in the anxiety-related dimensions were higher in S 12 than in L 12 carriers. Opposite in direction, scores tended to be lower in S 10 than in L 10 carriers. In the novelty seeking-related dimensions, scores were higher in S 10 than in S 12 carriers. No association was observed in women. In conclusion, anxiety- and novelty seeking-related personality dimensions are differentially associated with different SERT haplotypes; the consistent restriction to men suggests common androgen regulation. Opposite trends with haplotypes including the same promoter alleles suggest contribution of group stratification to earlier inconsistent findings and call to further differentiate the molecular function and clinical implications of the SERT promoter polymorphism. PMID- 16313924 TI - Angiogenesis and inflammation in skeletal muscle in response to ascites tumor in mice. AB - This study addresses the interaction between Ehrlich ascites tumor and skeletal abdominal muscle, presenting quantitative analysis of ascites-induced angiogenesis and inflammation in this tissue of mice bearing-tumor. Time dependent changes in the muscle (cellular activity, angiogenesis, inflammation and cytokines production) were assessed by morphometric, functional, and biochemical parameters at days 1, 4 and 8 after i.p. inoculation of Ehrlich tumor cells (2.5 x 10(7)). The number of cells stained with AgNOR technique (argyrophilic nucleolar organizer region) in the muscle, together with MTS assay used as markers of cellular activity increased progressively in parallel with the out flow rate of sodium fluorescein (blood flow index), hemoglobin content (vascular index) and VEGF production. Likewise, the inflammatory process in the muscle, as assessed by myeloperoxidase (MPO) and n-acethylglucosaminidase (NAG) activities and the levels of the chemokines, keratinocyte-derived chemokine (CXC1 3/KC) and macrophage-chemoattractant protein (CCL2/MCP-1) increased with tumor development. The combination of techniques used to describe angiogenesis and inflammation in a muscle model system has proved to be suited for quantitative measurements of microvascular changes and cellular infiltration occurring in the abdominal muscle wall of ascites-bearing mice. This study holds potential for investigating events and mechanisms associated with skeletal muscle response to neoplasic stimulus. PMID- 16313925 TI - Preferential inhibition of human phosphodiesterase 4 by ibudilast. AB - Ibudilast ophthalmic solution exhibited an improved clinical efficacy over cromoglycate in the treatment of allergic conjunctivitis. To further characterize its principal mode of action, the phosphodiesterase (PDE) inhibitory profile of ibudilast has been examined using human recombinant enzymes. Ibudilast, but not the other commonly used anti-allergic ophthalmic solutions including cromoglycate, ketotifen, tranilast and levocabastine, potently inhibits purified human PDE4A, 4B, 4C and 4D with IC50 values at 54, 65, 239 and 166 nM, respectively. Ibudilast effectively blocks lipopolysaccharide (LPS)-induced tumor necrosis factor (TNFalpha, IC50 = 6.2 microM) and N-formyl-Met-Leu-Phe (fMLP) induced leukotriene (LT) B4 biosynthesis (IC50 = 2.5 microM) in human whole blood, which are 3 and 6-fold more potent than cilomilast, respectively. The attenuated inflammatory and allergic responses from the potent and preferential PDE4 inhibition of ibudilast may have contributed significantly to its beneficial pharmacological responses and distinguishes ibudilast from the other ophthalmic solutions in the treatment of ocular allergy. PMID- 16313926 TI - Hypolipidemic and antioxidant effects of Morus alba L. (Egyptian mulberry) root bark fractions supplementation in cholesterol-fed rats. AB - The 70% alcohol extract of the Egyptian Morus alba L. root bark was fractionated over cellulose CC eluted with water, 50% methanol and finally with 100% methanol to yield 3 fractions (MRBF-1, MRBF-2 and MRBF-3), respectively. In continuation of chromatographic purification of 70% alcohol extract fractions of the Egyptian M. alba L. root bark, 4 compounds namely: mulberroside A, 5,7,2' trihydroxyflavanone-4'-O-beta-D-glucoside and albanols A and B were isolated from MRBF-2 for the first time from the Egyptian plant. Experimentally induced atherosclerosis was produced by feeding rats a diet enriched in coconut oil (25% by weight) and cholesterol (2% by weight) for 21 days. Then, hypercholesterolemic rats were orally administered (MRBF-1, MRBF-2 and MRBF-3 fractions) in a dose of 500 mg kg(-1) day(-1) for 15 successive days, in order to evaluate their expected hypocholesterolemic activity. Lipid profile parameters such as plasma total cholesterol, LDL-C, VLDL-C, LDL:HDL ratio and triglycerides, as well as plasma and liver lipid peroxides and glutathione-S-transferase enzyme levels, serum paraoxonase enzyme level, LDL oxidation, LDL aggregation and LDL retention, were measured. Plasma and liver glutathione-S-transferase enzyme levels were unaffected in all studied groups. The results revealed that the administration of (MRBF-2 and/or MRBF-3) fractions resulted in alleviation of atherosclerotic state. Administration of MRBF-3 significantly retained plasma and liver peroxides towards their normal levels, and also, produced significant increase in resistance towards major atherogenic modifications; namely LDL oxidation, LDL aggregation and LDL retention by 44%, 30%, and 33%, respectively. Thus, it can be concluded that the consumption of MRBF-2 and (MRBF-3, in some extent) fractions of M. alba L. root bark 70% alcohol extract may act as a potent hypocholesterolemic nutrient and powerful antioxidant via the inhibition of LDL atherogenic modifications and lipid peroxides formation in hypercholesterolemic rats. PMID- 16313927 TI - Alterations of behavior and spatial learning after unilateral entorhinal ablation of rats. AB - The entorhinal cortex (EC) is the key input and output structure of the hippocampus. It plays a crucial role in sensory processing, memory and learning, as well as in mechanisms of epileptic seizures. Our previous studies on the 4 aminopyridin induced epilepsy model of rats showed that ablation of unilateral EC prompted weakening of limbic seizure manifestation, thus the possibility of therapeutical benefit of this kind of surgery can be risen. Open field, elevated plus-maze and Morris water-maze test were performed to analyze changes of the basal activity level, exploratory behavior, and spatial memory capacity, respectively, of adult Wistar rats having undergone left EC excision. Compared with the sham-operated control group, rats with lesions of the EC showed enhanced locomotor activity in the open-field test. The elevated plus-maze test revealed higher frequency of entries and more time spent in the open arms. Morris water maze test suggested impairment of the spatial learning capacity following left lateral EC lesion. Therefore, our data showed that EC lesions induced hyperactivity, increased exploratory behavior, and impaired spatial learning. Entorhinal cortex ablation, as a potential method for controlling epileptic seizures has multiple effects on animals' behavior and spatial learning. To determine the cost-benefit ratio of a potential surgical intervention needs further experimental and human investigations. PMID- 16313928 TI - The effect of IL-1alpha on the expression of matrix metalloproteinases, plasminogen activators, and their inhibitors in osteoblastic ROS 17/2.8 cells. AB - Interleukin-1 (IL-1) plays key roles in altering bone matrix turnover. This turnover is regulated by matrix metalloproteinases (MMPs), tissue inhibitor of matrix metalloproteinases (TIMPs), and the plasminogen activation system, including tissue-type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA) , and plasminogen activator inhibitor type-1 (PAI-1). In this study, we examined the effect of IL-1alpha on the expression of the MMPs, TIMPs, tPA, uPA, and PAI-1 genes in osteoblasts derived from the rat osteosarcoma cell line ROS 17/2.8. The cells were cultured in alpha-minimum essential medium containing 10% fetal bovine serum with 0 or 100 U/ml of IL-1alpha for up to 14 days. The levels of MMPs, TIMPs, uPA, tPA, and PAI-1 expression were estimated by determining the mRNA levels using real-time RT-PCR and by determining protein levels using ELISA. In IL-1alpha cultures, the expression levels of MMP-1, -2, 3, -13, and -14 exceeded that of the control through day 14 of culture, and the expression of MMPs increased markedly from the proliferative to the later stages of culture. The TIMP-1, -2, and -3 expression levels increased from the initial to the proliferative stages of culture. The expression of tPA increased greatly during the proliferative stage of culture, and uPA expression increased throughout the culture period, increasing markedly from the proliferative to the later stages of culture. In contrast, PAI-1 expression decreased in the presence of IL-1alpha through day 14. These results suggest that IL-1alpha stimulate bone matrix turnover by increasing MMPs, tPA, and uPA production and decreasing PAI-1 production by osteoblasts, and incline the turnover to the resolution. PMID- 16313929 TI - The effects of metal distribution and anthropogenic effluents on the benthic environment of Gwangyang Bay, Korea. PMID- 16313930 TI - Dyslexics are impaired on implicit higher-order sequence learning, but not on implicit spatial context learning. AB - Developmental dyslexia is characterized by poor reading ability and impairments on a range of tasks including phonological processing and processing of sensory information. Some recent studies have found deficits in implicit sequence learning using the serial reaction time task, but others have not. Other skills, such as global visuo-spatial processing may even be enhanced in dyslexics, although deficits have also been noted. The present study compared dyslexic and non-dyslexic college students on two implicit learning tasks, an alternating serial response time task in which sequential dependencies exist across non adjacent elements and a spatial context learning task in which the global configuration of a display cues the location of a search target. Previous evidence indicates that these implicit learning tasks are based on different underlying brain systems, fronto-striatal-cerebellar circuits for sequence learning and medial temporal lobe for spatial context learning. Results revealed a double dissociation: dyslexics showed impaired sequence learning, but superior spatial context learning. Consistent with this group difference, there was a significant positive correlation between reading ability (single real and non word reading) and sequence learning, but a significant negative correlation between these measures and spatial context learning. Tests of explicit knowledge confirmed that learning was implicit for both groups on both tasks. These findings indicate that dyslexic college students are impaired on some kinds of implicit learning, but not on others. The specific nature of their learning deficit is consistent with reports of physiological and anatomical differences for individuals with dyslexia in frontal and cerebellar structures. PMID- 16313931 TI - Visuospatial deficits in patients with early left-hemispheric lesions and functional reorganization of language: consequence of lesion or reorganization? AB - INTRODUCTION: Patients with early left-hemispheric lesions have repeatedly been reported to show spared language functions (which are then mediated by the right hemisphere), but exhibit deficits in visuospatial functions. The "crowding hypothesis" explains these deficits by a neuronal scarcity in the right hemisphere for the original right hemispheric functions. An alternative hypothesis suggests direct lesion effects as the sole reason for the visuospatial impairments. The scope of this study was to examine the relations between visuospatial skills and hemispheric preference for language, and between visuospatial skills and cerebral lesion size. METHODS: In a sample of young adults with pre- or perinatally acquired focal lesions of the left hemisphere and unilateral spastic cerebral palsy on the right side, we assessed intelligence and neuropsychological functions in the verbal and visuospatial domains. The behavioural data was correlated with structural MRI information and language preference as assessed using fMRI. RESULTS: Patients with right hemispheric language production were impaired in visuospatial functions, compared not only to normal controls, but also to patients without right hemispheric language preference. Additionally, the degree of right hemispheric language correlated negatively with performance in visuospatial tasks. Lesion size correlated negatively only with performance IQ, which again correlated negatively with motor impairment of the patients and thus does not seem to reflect true cognitive deficits. CONCLUSION: Visuospatial deficits in patients with early left hemispheric lesions are a consequence of lesion-induced right hemispheric language organization, thus lending further support to the "crowding hypothesis". PMID- 16313932 TI - Pulmonary artery wedge pressure during treadmill exercise in warmblood horses with atrial fibrillation. AB - The heart rate and the pulmonary artery wedge pressure (PWP) was measured in 10 healthy warmblood horses and in six warmblood horses with atrial fibrillation (AF) at rest and during standardised treadmill exercise. During treadmill exercise, the increase in heart rate was significantly higher in the horses with AF than in the healthy horses. Horses with AF showed a significantly higher increase in PWP at treadmill velocities of 5m/s and faster, than did the healthy horses. The differences in PWP between both groups increased with treadmill strain. The present study demonstrates that there is an influence on the haemodynamics in horses with AF during treadmill exercise, which could explain exercise intolerance in some horses with lone AF. PMID- 16313933 TI - Persistent Epstein-Barr viral reactivation in young African children with a history of severe Plasmodium falciparum malaria. AB - Epstein-Barr virus (EBV) and Plasmodium falciparum have overlapping distributions and are thought to have causal interactions, particularly with regard to the aetiology of endemic Burkitt's lymphoma. Using real-time PCR, we quantified and compared EBV DNA levels in the blood before and after antimalarial treatment of age- and gender-matched groups of Gabonese children who presented with either mild or severe P. falciparum malaria. Following treatment, the prevalence of EBV DNA declined in the mild malaria group but increased in the severe malaria group, and a significantly higher proportion of the latter had EBV DNA detectable in their blood when they were healthy and parasite free (67% vs. 39%; P=0.013). High EBV DNA loads were associated with more malaria attacks and with elevated plasma concentrations of both TNF-alpha and IL-12p40. Significantly more under 5 year olds had EBV DNA, highlighting the strong age dependence of the interaction between the two pathogens. These findings confirm that EBV is reactivated during acute P. falciparum malaria but, importantly, also reveal that: (i) EBV activity persists at a higher frequency in children with a history of severe malaria; and (ii) higher peripheral blood EBV DNA loads are associated with susceptibility to more frequent P. falciparum episodes and with altered cytokine activity. PMID- 16313934 TI - Poor performance status is associated with early death in patients with pulmonary tuberculosis. AB - The objective of this study was to determine whether poor performance status at the start of anti-tuberculous (anti-TB) treatment is associated with early death in patients admitted to hospital with pulmonary tuberculosis (PTB). During 3 months in 2001, all adult patients admitted to eight hospitals in Limpopo Province, South Africa, and diagnosed with PTB were eligible for inclusion. At initiation of anti-TB treatment, a performance status between 0 and 4 was estimated for each patient using a modified version of the Eastern Cooperative Oncology Group scoring system. Hospital records and local TB registers were reviewed to identify patients who had died during the first 2 months of treatment. In addition, it was ascertained whether a death notification had been received by the provincial administration. Fifty-three of 295 (18%) patients died within 2 months. Mortality increased from 6% in patients with the best performance status to 51% in patients with the poorest performance status. Univariate and multivariate Cox regression analysis showed that the hazard ratio for dying was significantly higher for patients with a performance status of 3 or 4. Poor performance status shows a strong association with early death in patients with PTB and has the potential to be a useful clinical, epidemiological and research tool. PMID- 16313935 TI - Efficacy of chloroquine-proguanil malaria prophylaxis in a non-immune population in Bangui, Central African Republic: a case-control study. AB - A case-control study was conducted to evaluate the efficacy of the combination of chloroquine plus proguanil as malaria prophylaxis in a non-immune population living in the Central African Republic. Cases were patients presenting with a malaria attack confirmed by a positive blood film and/or an HRP2 positive antigen test at the Pasteur Institute of Bangui. Two control subjects were included per case: one was a relative or close friend and the other was matched to the patient with respect to the length of stay. A questionnaire assessing malaria prophylaxis habits and malarial risk factors over the 2-month period prior to inclusion in the study was given to 48 cases and 96 controls. A conditional logistic regression was used to identify risk factors. The efficacy of the chloroquine plus proguanil regimen was found to be high (95.5%, 95% CI 74.0-99.2%) in this country known for high chloroquine resistance. Our data lend some support to the use of chloroquine plus proguanil in Bangui, and the protective efficacy of chloroquine plus proguanil should now be studied prospectively as part of a randomised controlled trial of various prophylactic drugs. PMID- 16313936 TI - Comparison of sound fields generated by different coded excitations--experimental results. AB - This work reports the results of measurements of spatial distributions of ultrasound fields obtained from five energizing schemes. Three different codes, namely, chirp signal and two sinusoidal sequences were investigated. The sequences were phase modulated with 13 bits Barker code and 16 bits Golay complementary codes. Moreover, two reference signals generated as two and sixteen cycle sine tone bursts were examined. Planar, 50% (fractional) bandwidth, 15 mm diameter source transducer operating at 2 MHz center frequency was used in all measurements. The experimental data were collected using computerized scanning system and recorded using wideband, PVDF membrane hydrophone (Sonora 804). The measured echoes were compressed, so the complete pressure field in the investigated location before and after compression could be compared. In addition to a priori anticipated increase in the signal to noise ratio (SNR) for the decoded pressure fields, the results indicated differences in the pressure amplitude levels, directivity patterns, and the axial distance at which the maximum pressure amplitude was recorded. It was found that the directivity patterns of non-compressed fields exhibited shapes similar to the patterns characteristic for sinusoidal excitation having relatively long time duration. In contrast, the patterns corresponding to compressed fields resembled those produced by brief, wideband pulses. This was particularly visible in the case of binary sequences. The location of the maximum pressure amplitude measured in the 2 MHz field shifted towards the source by 15 mm and 25 mm for Barker code and Golay code, respectively. The results of this work may be applicable in the development of new coded excitation schemes. They could also be helpful in optimizing the design of imaging transducers employed in ultrasound systems designed for coded excitation. Finally, they could shed additional light on the relationship between the spatial field distribution and achievable image quality and in this way facilitate optimization of the images obtained using coded systems. PMID- 16313937 TI - Virus replication and disease progression inversely correlate with SIV tat evolution in morphine-dependent and SIV/SHIV-infected Indian rhesus macaques. AB - We analyzed the association between evolution of the 5' exon of tat and disease progression in an SIV/SHIV macaque model of opiate dependence and AIDS. Cloned tat sequences were obtained by RT-PCR amplification of 3 plasma viruses (recovered at different times) from 6 morphine-dependent and 2 control Indian rhesus macaques inoculated with SHIV(KU-1B) SHIV(89.6P) and SIV/17E-Fr. Approximately ten clones were sequenced for each animal per time point for use in phylogenetic analyses. We found a strong, significant inverse correlation between disease progression and tat diversity in plasma by 20 weeks post-infection. The morphine-dependent macaques developed 2 distinct disease patterns - rapid progressor (Group A) and slow progressor (Group B) - whereas control animals developed into slow progressor only (Group C). The three animals in Group A exhibited approximately 40% (P = 0.01) and approximately 50% (P = 0.028) less diversity than Group B and C animals, respectively, over the 20 weeks. Furthermore, the Group A macaques showed a prominent reemergence of the wild-type SV17E tat sequence used in the inoculum that coincided with disease progression. This suggests that the virus from the original infection represented the most pathogenic form among all animals in these cohorts throughout the first 20 weeks of infection. We were unable to support or rule out a role for immune pressure on tat evolution based on the spectrum of sequence changes in the data set. Thus, in the short duration of this study, the Tat-specific immune pressure cannot explain the different disease outcomes of the six morphine animals nor of the two controls. Our results also suggest that in vivo morphine dependence can contribute to the pathogenesis of SIV/SHIV infection and that it may do so in conjunction with the evolution of viral proteins, such as Tat. PMID- 16313938 TI - Molecular identification and phylogenetic analysis of baculoviruses from Lepidoptera. AB - PCR amplification of the highly conserved baculovirus genes late expression factor 8 (lef-8), late expression factor 9 (lef-9) and polyhedrin/granulin (polh/gran) combined with molecular phylogenetic analyses provide a powerful tool to identify lepidopteran-specific baculoviruses and to study their diversity. In the present investigation, we have improved the degenerate oligonucleotides and corroborated the approach that was recently described by Lange et al. (Lange, M., Wang, H., Zhihong, H., Jehle, J.A., 2004. Towards a molecular identification and classification system of lepidopteran-specific baculoviruses. Virology 325, 36 47.). Baculovirus DNA was isolated from 71 uncharacterized historic baculovirus samples, and partial gene sequences were amplified by using gene-specific degenerate PCR primers. The obtained PCR products were directly sequenced, and the deduced amino acid sequences were compiled and aligned with published sequences of these target genes. A phylogenetic tree of 117 baculoviruses was inferred using maximum parsimony and distance methods. Based on the comprehensive phylogenetic analysis of the partial lef-8, lef-9 and polh/gran genes, we propose a phylogenetic species criterion for lepidopteran-specific baculoviruses that uses the genetic distances of these genes for species demarcation. PMID- 16313939 TI - Modelling nitrification, heterotrophic growth and predation in activated sludge. AB - A mathematical model describing the interaction between nitrifiers, heterotrophs and predators in wastewater treatment has been developed. The inclusion of a predation mechanism is a new addition to the existing activated sludge models. The developed model considered multi-substrate consumption and multi-species growth, maintenance and decay in a culture where nitrifiers, heterotrophs and predators (protozoa and metazoa) are coexisting. Two laboratory-scale sequenced batch reactors (SBRs) operated at different sludge retention time (SRT) of 30 and 100 days for a period of 4 years were used to calibrate and validate the model. Moreover, to assess the predator activity, a simple procedure was developed, based on measuring the respiration rate with and without the presence of the predators. The model successfully described the performance of two SBRs systems. The fraction of active biomass (ammonia oxidisers, nitrite oxidisers and heterotrophs) predicted by the proposed model was only 33% and 14% at SRT of 30 and 100 days, respectively. The high fraction of inert biomass predicted by the model was in accordance with the microscopic investigations of biomass viability in both reactors. The presented model was used to investigate the effect of increasing sludge age and the role of predators on the biomass composition of the tested SBR system. PMID- 16313940 TI - Coupling of sequencing batch reactor and mesh filtration: operational parameters and wastewater treatment performance. AB - Wastewater treatment performance of the combined process of sequencing batch reactor (SBR) and mesh filtration bio-reactor was investigated with a synthetic wastewater. In this system, the filtration was performed only by the water level difference between the reactor and the effluent port, with the help of a sludge layer which accumulated on the mesh filter. A half volume of the mixed liquor was filtrated for ca. 1 h, and the filtration time was not affected by the initial pressure within the range of 0.5-2.0 m-H2O. Since the mesh filter could effectively reject the biomasses in the reactor, the effluents contained SS of less than 1 mg/L and BOD of less than 10 mg/L under continuous or intermittent aeration conditions. Nitrogen was also removed effectively with the adjustment of aeration time under the intermittent aeration conditions. The results obtained in this work indicate that mesh filtration could be effectively combined with SBR and improve the performance of SBR. In addition, it was shown that the performance of the mesh filtration such as filtration time and solids separation was influenced significantly by the saccharide content in the exocellular polymer of the activated sludge. PMID- 16313941 TI - Comparison of commercially available Escherichia coli enumeration tests: implications for attaining water quality standards. AB - Many states are replacing microbiological water quality standards based on "fecal" or thermotolerant coliforms (ThCs) with new standards that employ Escherichia coli as the indicator organism. Implicit in these new standards are assumptions about the equivalence of E. coli enumeration tests and the E. coli levels that will provide protection equivalent to former ThC standards. To investigate these assumptions, E. coli levels in split samples (tests conducted on portions of the same grab sample) collected from small urban streams were determined using enzyme-specific media (Colilert, m-ColiBlue24, and nutrient agar with MUG) and compared to levels determined with conventional culture media (m-FC and m-TEC). Although levels observed with all tests were highly correlated, significantly fewer E. coli were enumerated with m-TEC than with enzyme-specific media (paired Student's t-test, alpha>99%). In addition, E. coli were found to comprise a larger fraction (84-104%, depending on the test) of the total presumptive ThC concentration than that suggested by the United States Environmental Protection Agency (63%). Both of these observations (1) the improvement in E. coli yields observed with enzyme-specific media, and (2) the greater proportion of ThC organisms that are E. coli, indicate that more water quality violations will occur when enzyme-specific media are used for testing than if conventional culture media are used. PMID- 16313942 TI - Microbiological investigations for sanitary assessment of wastewater treated in constructed wetlands. AB - Microbiological investigations into the occurrence and fate of pathogens in wastewater were carried out in a multi-annual measurement programme, using samples from three artificial wetlands equipped with pre-treatment systems (multi chamber septic tanks, lagoons) and predominantly treating domestic wastewater. In this study, the concentrations of indicator organisms and pathogenic or potentially pathogenic microorganisms were determined in the various components or stages of the systems. The evaluation of data from some 3.600 microbiological analyses and a comparison with older data from a plant which has already been operation for 18 y made it possible for the first time ever to include operational factors in the assessment. Average removal of all major indicator organisms and pathogens is 1.5-2.5 log units for one-stage systems and increases to 3-5 log units for multi-stage systems. Significant differences between horizontal and vertical filters were not found. If no clogging problems arise during their operation, multi-stage systems can meet the requirements of the EU bathing water and irrigation water directives. Influent concentration, wastewater temperature, and hydraulic loading rate were found to be major factors of influence. The performance of constructed wetlands in removing microorganisms is clearly superior to that of conventional biological activated sludge systems. PMID- 16313943 TI - Determination of trichloroacetic acid in environmental studies using carbon 14 and chlorine 36. AB - Radioisotopes carbon 14 and chlorine 36 were used to elucidate the environmental role of trichloroacetic acid (TCA) formerly taken to be a herbicide and a secondary air pollutant with phytotoxic effects. However, use of 14C-labeling posed again known analytical problems, especially in TCA extraction from the sample matrix. Therefore--after evaluation of available methods--a new procedure using decarboxylation of [1,2-14C]TCA combined with extraction of the resultant 14C-chloroform with a non-polar solvent and its subsequent radiometric measurement was developed. The method solves previous difficulties and permits an easy determination of amounts between 0.4 and 20 kBq (10 - 500 ng g(-1)) of carrier-less [1,2-14C]TCA in samples from environmental investigations. The procedure is, however, not suitable for direct [36Cl]TCA determination in chlorination studies with 36Cl. Because TCA might be microbially degraded in soil during extraction and sample storage and its extraction from soil or needles is never complete, the decarboxylation method--i.e. 2 h TCA decomposition to chloroform and CO2 in aqueous solution or suspension in closed vial at 90 degrees C and pH 4.6 with subsequent CHCl3 extraction-is recommended here, estimated V < 7%. Moreover, the influence of pH and temperature on the decarboxylation of TCA in aqueous solution was studied in a broad range and its environmental relevance is shown in the case of TCA decarboxylation in spruce needles which takes place also at ambient temperatures and might amount more than 10-20% after a growing season. A study of TCA distribution in spruce needles after below-ground uptake shows the highest uptake rate into current needles which have, however, a lower TCA content than older needle-year classes, TCA biodegradation in forest soil leads predominatingly to CO2. PMID- 16313944 TI - Zea mays L. protein changes in response to potassium dichromate treatments. AB - The plant metabolic response to heavy metal stress is largely unknown. The present investigation was undertaken to examine the influence of different concentrations of potassium dichromate on the Zea mays L. plantlets. A clear effect of chromium on maize plantlets growth and seed germination was observed strating from 100-300 ppm up to 1500 ppm. In this concentration range, chromium uptake was dependent on the concentration in the medium. Metallothioneins, involved in heavy metal binding, were measured by capillary electrophoresis (CE), and showed a dose-response induction. Protein profile analyzed by two-dimensional gel electrophoresis showed differential expression of several proteins. Identification of spots of upregulated proteins was performed by MALDI mass spectrometry. Results showed that proteins induced by heavy metal exposure are principally involved in oxidative stress tolerance or in other stress pathways. Induction of proteins implicated in sugar metabolism was also observed. Identification of factors involved in plant response may lead to a better understanding of the mechanisms involved in cell protection and tolerance. This information could be used to improve agricultural production and environmental quality. PMID- 16313945 TI - CD8 T-cell immune phenotype of successful aging. AB - The nonagenarian population by definition represents individuals who have demonstrated success in aging. We determined the status of CD8(+) T-cell senescence in nonagenarians by analyzing the expression of CD28 and Fas (CD95), and analyzing activation and activation-induced cell death (AICD). Peripheral blood mononuclear cells (PBMCs) were isolated from three groups of subjects: adults (20-64-year-old), older adults (65-89-year-old), and nonagenarians (>or=90 year-old). PBMCs were stimulated with phytohemagglutinin (PHA) (10 microg/ml). The cells were labeled with conjugated antibodies specific for CD4, CD8, CD28, CD45RO, and Fas, and were analyzed by FACS((R)). There was a strong negative correlation of the percentage of CD28(+)Fas(-) CD8(+) T-cells with the age of each individual prior to stimulation in vitro (R(2)=0.76, p<0.0001). Compared to other biomarkers (CD28(-), CD28(-)CD45RO(+), and Fas(+)) that have been associated with CD8(+) T-cell aging, the loss of the CD28(+)Fas(-) CD8(+) T-cell population exhibited the strongest correlation with the individual's chronologic age. After stimulation with PHA, there was a decrease in the percentage of CD8(+) T-cells from individual >or=65-year-old that expresses both CD28(+) and Fas(+) at day 3. Surprisingly, the AICD response of CD8(+) T-cells at day 7 in the nonagenarians was higher than that in the other two groups. These results suggest that successful aging does not prevent development of the senescent phenotype of unstimulated CD8(+) T cells, but is associated with preservation of CD8 T cell functions including activation and AICD. Increased AICD may result in enhanced rejuvenation capacity of T cells and limit the impact of aging on T cell function in nonagenarians. PMID- 16313946 TI - Factors influencing development of management strategies for the Abou Ali River in Lebanon I: spatial variation and land use. AB - Surface water bodies are progressively subject to increasing stress as a result of environmentally degrading processes primarily related to anthropogenic activities. This study assesses and examines the impact of land use and land based activities on the spatial variation in water quality of the Abou Ali River in North Lebanon. It is the first detailed study of its kind in Lebanon and adds to the existing knowledge by shedding light on a relatively small Mediterranean river in a developing country where there is a paucity of such studies. The assessment was conducted at the end of the dry season in 2002 and 2003 and the end of the wet season in 2003 and 2004. The study has demonstrated the importance of anthropogenic influences on the water quality of the Abou Ali River Basin, as concentrations of most contaminants were higher at locations with greatest human activity. The most adversely affected area was the section of the river that flows through an entirely urbanized and highly populated region, the Tripoli conurbation. Upstream rural sites were enriched by contaminants primarily from non-point sources such as agricultural runoff and poultry litter whereas contaminant concentrations at the urban sites were enriched by a combination of sewage discharge and flow of contaminants from upstream. If the Abou Ali River is to be utilized as a managed water resource and its water quality sustained, point source discharges will require treatment and land use management must be planned to minimize the impact of diffuse source pollution on the river. A high priority should be given to the implementation and enforcement of the precautionary and polluter pays principles. Moreover, an effective legal, economic and institutional framework is required to encourage investment in waste reduction and control and to introduce environmentally sound practices. PMID- 16313947 TI - Occurrence of antibiotics in hospital, residential, and dairy effluent, municipal wastewater, and the Rio Grande in New Mexico. AB - This study had three objectives: 1) determine occurrence of antibiotics in effluent from hospitals, residential facilities, and dairies, and in municipal wastewater 2) determine antibiotic removal at a large wastewater treatment plant (WWTP) in Albuquerque, NM, and 3) determine concentrations of antibiotics in the Rio Grande, which receives wastewater from the Albuquerque WWTP. Twenty-three samples of wastewater and 3 samples of Rio Grande water were analyzed for the presence of 11 antibiotics. Fifty-eight percent of samples had at least one antibiotic present while 25% had three or more. Hospital effluent had detections of sulfamethoxazole, trimethoprim, ciprofloxacin, ofloxacin, lincomycin, and penicillin G, with 4 of 5 hospital samples having at least one antibiotic detected and 3 having four or more. At the residential sampling sites, ofloxacin was found in effluent from assisted living and retirement facilities, while the student dormitory had no detects. Only lincomycin was detected in dairy effluent (in 2 of 8 samples, at 700 and 6600 ng/L). Municipal wastewater had detections of sulfamethoxazole, trimethoprim, ciprofloxacin, and ofloxacin, with 4 of 6 samples having at least one antibiotic present and 3 having 3 or more. The relatively high concentrations (up to 35,500 ng/L) of ofloxacin found in hospital and residential effluent may be of concern due to potential genotoxic effects and development of antibiotic resistance. At the Albuquerque WWTP, both raw wastewater and treated effluent had detections of sulfamethoxazole, trimethoprim, and ofloxacin, at concentrations ranging from 110 to 470 ng/L. However, concentrations in treated effluent were reduced by 20% to 77%. No antibiotics were detected in the Rio Grande upstream of the Albuquerque WWTP discharge, and only one antibiotic, sulfamethoxazole, was detected in the Rio Grande (300 ng/L) below the WWTP. PMID- 16313948 TI - Bioreactivity of particulate matter in Beijing air: results from plasmid DNA assay. AB - An in vitro plasmid assay was employed to study the bioreactivity of PM (particulate matter) in Beijing air. It was found that the TD20 (toxic dose of PM causing 20% of plasmid DNA damage) of Beijing PM can be as low as 28 microg ml( 1) and as high as >1000 microg ml(-1). Comparison of the physical properties, such as morphology and size distribution, and oxidative potential indicates that the PM(2.5) (particulate matter with an aerodynamic diameter of 2.5 microm or less) has a stronger oxidative capacity than PM(10) (particulate matter with an aerodynamic diameter of 10 microm or less), and that the higher number percentages of soot aggregates and lower number percentages of mineral and fly ashes are associated with the higher oxidative capacity. Although the mass of PM(10) during dust storms is commonly 5 times higher than that during non-dust storm episodes, the oxidative capacity of PM(10)s of dust storms is much lower than that of the non-dust storm PM(10)s. The water-soluble fractions and intact whole particle solutions of Beijing airborne particles produce similar plasmid assay results, demonstrating that the bioreactivity of Beijing airborne particles is mainly sourced from the water-soluble fraction. In the samples with stronger bioreactivity, the total analyzed water soluble Al, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As and Pb (ppm) concentrations are higher. The water soluble zinc shows a good negative correlation with TD20s, suggesting that the water-soluble zinc is probably the major element responsible for the plasmid DNA damage. PMID- 16313949 TI - Warfarin anticoagulation in children made easier. PMID- 16313950 TI - Role of dopamine and glutamate receptors in cocaine-induced social effects in isolated and grouped male OF1 mice. AB - Cocaine administration in paired male mice decreases social contacts as well as increases avoidance and flee elements. As dopamine (DA) and glutamate seem to be involved in some of cocaine's effects, an attempt was made to assess whether a range of associated receptors influenced the social impacts of this drug of abuse. The NMDA antagonist memantine (10 and 40 mg/kg); the AMPA antagonist CNQX (1 and 20 mg/kg); the DA release inhibitor CGS 10746b (2 and 8 mg/kg): the DA D1 antagonist SCH 23390 (0.05 and 0.5 mg/kg); and the DA D2/D3 antagonist raclopride (0.03 and 0.3 mg/kg) were administered prior to 25 mg/kg of cocaine and behaviour was evaluated during an encounter between an experimental and a standard opponent in a neutral cage for 10 min. Memantine reverts cocaine-induced social withdrawal and the increase in avoidance and flee, CNQX being effective only in these latter actions. On the other hand, SCH 23390 counteracts the social as well as the defensive action of cocaine, raclopride being effective only in blocking the cocaine-induced increase in avoidance and flee behaviours. In conclusion, although both neurotransmitter systems are involved in the effects of cocaine on social behaviour, NMDA and D1DA receptors seem to have an important role. PMID- 16313951 TI - Osteoblast adhesion and matrix mineralization on sol-gel-derived titanium oxide. AB - The biological events occurring at the bone-implant interface are influenced by the topography, chemistry and wettability of the implant surface. The surface properties of titanium alloy prepared by either surface sol-gel processing (SSP), or by passivation with nitric acid, were investigated systematically using X-ray photoelectron spectroscopy, scanning electron microscopy, atomic force microscopy and contact angle metrology. The bioreactivity of the substrates was assessed by evaluating MC3T3-E1 osteoblastic cell adhesion, as well as by in vitro formation of mineralized matrix. Surface analysis of sol-gel-derived oxide on Ti6Al4V substrates showed a predominantly titanium dioxide (TiO(2)) composition with abundant hydroxyl groups. The surface was highly wettable, rougher and more porous compared to that of the passivated substrate. Significantly more cells adhered to the sol-gel-coated surface, as compared with passivated surfaces, at 1 and 24h following cell seeding, and a markedly greater number of mineralized nodules were observed on sol-gel coatings. Collectively our results show that the surface properties of titanium alloy can be modified by SSP to enhance the bioreactivity of this biomaterial. PMID- 16313952 TI - The effect of glow discharge plasma surface modification of polymers on the osteogenic differentiation of committed human mesenchymal stem cells. AB - Little is known of the effect of material surfaces on stem cell differentiation. The present study has addressed the hypothesis that the interaction of mesenchymal stem cells (MSCs) with material surfaces modified by glow discharge plasma is a major regulator of osteogenic differentiation. We found that biaxially oriented polypropylene (BOPP) plasma treated in ammonia significantly reduced up-regulation of expression of osteogenic marker genes, such as alkaline phosphatase (ALP), bone sialoprotein (BSP) and osteocalcin (OC). In contrast, ALP expression was up-regulated when cultured on treated Nylon-6 polyamide (Ny-t) but was substantially reduced when cultured on its pristine counterpart (Ny-p) on day 3. On day 7, ALP expression was down-regulated with MSCs cultured on Ny-t although its expression level was up again on day 14. BSP was expressed weakly on day 3, but was up-regulated when cultured on Ny-t and Ny-p. Its expression reached its maximum on day 14 when cultured on a polystyrene control, while it was cyclically up-regulated on Ny-t. Similarly, there was a slight increase in OC expression when MSCs were cultured on Ny-t and Ny-p on day 3, when compared to control. Thus, the nature of the surface can directly influence MSCs differentiation, ultimately affecting the quality of new tissue formation with BOPP-t suppressing osteogenic differentiation. PMID- 16313953 TI - In vitro and in vivo studies on vitamin E TPGS-emulsified poly(D,L-lactic-co glycolic acid) nanoparticles for paclitaxel formulation. AB - This work shows a full spectrum of research on Vitamin E TPGS-emulsified Poly(D,L lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) for paclitaxel formulation to improve its therapeutic index and to reduce the adverse effects of adjuvant Cremophor EL in its current clinical formulation of Taxol. Paclitaxel-loaded PLGA NPs were prepared by a modified solvent extraction/evaporation technique with vitamin E TPGS as emulsifier. The formulated NPs were found in quite uniform size of approximately 240 nm diameter. The in vitro drug release profile exhibited a biphasic pattern with an initial burst followed by a sustained release. In vitro HT-29 cell viability experiment demonstrated that the drug formulated in the NPs was 5.64, 5.36, 2.68, and 1.45 times more effective than that formulated in the Taxol formulation after 24, 48, 72, 96 h treatment, respectively at 0.25 microg/mL drug concentration, which should be even better with the sustainable release feature of the NPs formulation considered. In vivo PK measurement confirmed the advantages of the NP formulation versus Taxol. The area-under-the curve (AUC) for 48 h for Vitamin E TPGS emulsified PLGA NP formulation of paclitaxel were found 3.0 times larger than that for the Taxol formulation. The sustainable therapeutic time, at which the drug concentration drops below the minimum effective value, for the NP formulation could be 1.67 times longer than that for the Taxol formulation. PMID- 16313954 TI - Highly effective and slow-biodegradable network-type cationic gene delivery polymer: small library-like approach synthesis and characterization. AB - Over the past years, macromolecular biodegradable polymers have been attracting considerable attention as gene delivery vehicles due to their safety and many potential applications. In the process of developing such biopolymers, we synthesized a biodegradable, network-type poly(amino ester) polymer (nt-PAE), which showed desirable chemical properties and a mechanical durability with high transfection efficiency. A small library-like approach to polymer synthesis using melt polycondensation followed by surface modification with aminohexanoic acid resulted in the final polymer, nt-PAE. The network-type structure of nt-PAE consists of biodegradable ester linkages and tertiary amines embedded in the backbone, and primary amines positioned at the surface. Unlike other rapidly degrading ester polymers, the nt-PAE exhibited fairly slow degradation, as the polymer sustained its DNA complexing ability for 10 days under physiological buffer conditions. In addition, the transfection efficiency of the nt-PAE could be increased to the same level comparable with that of PEI even in the presence of serum. Low toxicity and high transfection efficiency with a slow degradation profile implies that the nt-PAE can be used as a valuable nonviral gene delivery system. PMID- 16313955 TI - A collagen-glycosaminoglycan co-culture model for heart valve tissue engineering applications. AB - In order to develop efficient design strategies for a tissue-engineered heart valve, in vivo and in vitro models of valvular structure and cellular function require extensive characterisation. Collagen and glycosaminoglycans (GAGs) provide unique functional characteristics to the heart valve structure. In the current study, type I collagen-GAG hydrogels were investigated as biomaterials for the creation of mitral valve tissue. Porcine mitral valve interstitial cells (VICs) and endothelial cells (VECs) were isolated and co-cultured for 4 weeks in hydrogel constructs composed of type I collagen. The metabolic activity and tissue organisation of mitral valve tissue constructs was evaluated in the presence and absence of chondroitin sulphate (CS) GAG, and comparisons were made with normal mitral valve tissue. Both collagen and collagen-CS mitral valve constructs contracted to form tissue-like structures in vitro. Biochemical assay demonstrated that over 75% of CS was retained within collagen-CS constructs. Morphological examination demonstrated enhanced VEC surface coverage in collagen CS constructs compared to collagen constructs. Ultrastructural analysis revealed basement membrane synthesis and cell junction formation by construct VECs, with an increased matrix porosity observed in collagen-CS constructs. Immunohistochemical analyses demonstrated enhanced extracellular matrix production in collagen-CS constructs, including expression of elastin and laminin by VICs. Both native valve and collagen-CS construct VECs also expressed the vasoactive molecule, eNOS, which was absent from collagen construct VECs. The present study demonstrates that collagen gels can be used as matrices for the in vitro synthesis of tissue structures resembling mitral valve tissue. Addition of CS resulting in a more porous model was shown to positively influence the bioactivity of seeded valve cells and tissue remodelling. Collagen-GAG matrices may hold promise for a potential use in heart valve tissue engineering and improved understanding of heart valve biology. PMID- 16313956 TI - A gold nanoparticles/sol-gel composite architecture for encapsulation of immunoconjugate for reagentless electrochemical immunoassay. AB - A highly hydrophilic, non-toxic and conductive colloidal gold nanoparticle/titania sol-gel composite membrane with a low contact angle was prepared on a glassy carbon electrode via a vapor deposition method. With human chorionic gonadotrophin (hCG) as a model antigen and encapsulation of horseradish peroxidase-labeled hCG antibody (HRP-anti-hCG) in the composite architecture, this membrane could be used for reagentless electrochemical immunoassay. It displayed a porous and homogeneous composite architecture without the aggregation of the immobilized protein molecules. The presence of gold nanoparticles provided a congenial microenvironment for adsorbed biomolecules and decreased the electron transfer impedance, leading to a direct electrochemical behavior of the immobilized HRP. The formation of immunoconjugate by a simple one-step immunoreaction between hCG in sample solution and the immobilized HRP-anti-hCG introduced a barrier of direct electrical communication between the immobilized HRP and the electrode surface. Under optimal conditions, the hCG analyte could be determined in two linear ranges from 0.5 to 5.0 mIU/mL and 5.0 to 30 mIU/mL with a relatively low detection limit of 0.3 mIU/mL at 3sigma. The hCG immunosensor exhibited good precision, high sensitivity, acceptable stability, accuracy and reproducibility. This composite membrane could be used efficiently for the entrapment of different biomarkers and clinical applications. PMID- 16313957 TI - Designing pregnancy centered medications: drugs which do not cross the human placenta. AB - This review considers and evaluates the role of placental transporters (multidrug resistance proteins, P-glycoprotein and breast cancer resistance protein) in the uptake and efflux of drugs used in pregnancy. The effect of placental transporters in effluxing drugs such as glyburide and numerous protease inhibitors from the fetal circulation offers the potential to manipulate the passage of drugs across the placenta. The discovery of the interactions of these drugs with placental transporters may provide a novel framework for future drug development in which medications can be designed to control the degree of fetal exposure and thus prevent fetal risk. PMID- 16313958 TI - Dynorphin A (2-13) improves mecamylamine-induced learning impairment accompanied by reversal of reductions in acetylcholine release in rats. AB - Accumulating evidence indicates that the endogenous opioid peptides dynorphin A (1-17) and synthetic dynorphin A (1-13) interact not only with opioid receptors but also with as yet poorly characterized non-opioid binding sites. Dynorphin A (1-13) improved impairments of learning and memory via not only kappa-opioid receptor-mediated, but also 'non-opioid' mechanisms. In the present study, the effects of des-tyrosine(1) dynorphin A (2-13) as a non-opioid metabolite of dynorphin A, and dynorphin A (1-13) on mecamylamine-induced impairment of the acquisition of learning in rats were investigated using a step-through type passive avoidance task. Further, hippocampal acetylcholine release was examined using in vivo microdialysis. Mecamylamine significantly shortened the step through latency when given 30 min before the acquisition trial. Not only dynorphin A (1-13) but also dynorphin A (2-13) attenuated the mecamylamine induced impairment of the acquisition of learning. The effect of dynorphin A (2 13) was not blocked by pre-treatment with nor-binaltorphimine (nor-BNI), a selective kappa-opioid receptor antagonist. Dynorphin A (2-13) completely abolished the decrease in the extracellular acetylcholine concentration induced by mecamylamine and this effect was not blocked by nor-BNI. Taken together with our previous findings, the present results may indicate that dynorphin A (2-13) improves impairment of learning and/or memory in 'non-opioid' mechanisms and dynorphin A (1-13) ameliorates impairment of the acquisition of learning via not only kappa-opioid receptor-mediated mechanisms but also 'non-opioid' mechanisms, by regulating the release of extracellular acetylcholine. PMID- 16313959 TI - Changes in immunoglobulin-nucleoprotein complex structure mapped by chromatin immunoprecipitation. AB - Transcription factor-mediated immunoglobulin (Ig) enhancer activation has been analyzed extensively outside the physiological constraints of chromatin. Towards understanding the role sequence-specific DNA binding proteins identified by these methods play in activating Ig genes during B cell development, we have investigated in vivo interaction between the Ig enhancer activator PU.1 and two target elements, the Igmu and kappa3' enhancers, by chromatin immunoprecipitation (ChIP). By using two antibodies recognizing different PU.1 epitopes in murine B cells, these analyses demonstrate that ChIP results may depend on the availability of the epitope(s) targeted by the immunoprecipitating antibody. Specifically, PU.1 epitope availability at the mu and kappa3' enhancers does not accurately quantitate total PU.1 association. This result suggests the nucleoprotein complexes formed at these various active enhancers is cell type specific. Interestingly, RAG1-/- but not RAG2-/- pro-B cells lack PU.1/kappa3' association, probably due to limited accessibility of the kappa locus in the former. The more robust association of PU.1 with the kappa3' versus mu enhancer in all but RAG1-/- B lineage cells is not explained by differences in PCR primer efficiency, but likely reflects the different structures formed by the complexes at mu versus kappa3' enhancers. Finally, PU.1 is not associated with an inaccessible mu or kappa3' enhancer chromatin structure in macrophages, again emphasizing the importance cellular protein context plays in PU.1/Ig enhancer association. The demonstration that changes in epitope availability, hence nucleoprotein structure, can be monitored by ChIP suggests using this technique to monitor biologically important changes in nucleoprotein complex structure/composition in situ. PMID- 16313960 TI - Transcriptome response following administration of a live bacterial vaccine in Atlantic salmon (Salmo salar). AB - Antibacterial responses have been studied in Atlantic salmon following an acute intra peritoneal injection of a genetically attenuated (aroA(-)) strain of Aeromonas salmonicida known to elicit protective immunity. Three tissues were studied for transcriptional changes, the liver, head kidney and the gill. RNA was collected from fish 6, 12, 24 and 48 h following infection or at the same time points from fish injected with PBS as non-infected control. PCR-select cDNA subtraction libraries were constructed from pooled 24 and 48 h post infection RNA to identify up-regulated mRNAs. One thousand four hundred and eighty six cDNA clones were sequenced from enriched cDNA libraries, of which 71% had significant homologies to known functional proteins. Many of these clones have previously been un-characterised in Atlantic salmon. A salmonid cDNA microarray was used to further analyse the gene expression profile as the library construction in itself does not answer the dynamics of the response. The greatest increase in expression identified in the array analysis was a liver antibacterial peptide, hepcidin that was increased 11-fold following the challenge. A panel of clones were chosen for semiquantitative reverse transcriptase PCR from all time points sampled. These results indicated there were both temporal differences and tissue differences in the transcriptional response to bacterial exposure, potentially of relevance to the establishment of protection. PMID- 16313961 TI - Synthesis, spectroscopy (IR, multinuclear NMR, ESI-MS), diffraction, density functional study and in vitro antiproliferative activity of pyrazole-beta diketone dihalotin(IV) compounds on 5 melanoma cell lines. AB - Novel 4-acylpyrazolon-5-ato-dihalotin(IV) complexes, [Q2SnX2], (X = F, Cl, Br or I); HQ = HQ(CHPh2) (1,2-dihydro-3-methyl-1-phenyl-4-(2,2-diphenylacetyl)pyrazol-5 one), HQ(Bn) (1,2-dihydro-3-methyl-1-phenyl-4-(2-phenylacetyl)pyrazol-5-one) or HQ(CF3,py) (4-(2,2,2-trifluoroacetyl)-1,2-dihydro-3-methyl-1-(pyridin-2 yl)pyrazol-5-one) have been synthesized and characterized by spectroscopic (IR, 1H, 13C, 19F and 119Sn NMR, electrospray ionisation mass spectrometry (ESI-MS)), analytical and structural methods (X-ray and density functional theory). 119Sn chemical shifts depend on the nature of the halides bonded to tin. Isomer conversion, detected in solution by NMR spectroscopy, is related to the acyl moiety bulkiness while the cis(Cl)-cis(acyl)-trans(pyrazolonato) scheme is found in the solid state. The in vitro antiproliferative tests of three derivatives on three human melanoma cell lines (JR8, SK-MEL-5, MEL501) and two melanoma cell clones (2/21 and 2/60) show dose-dependent decrease of cell proliferation in all cell lines. The activity correlates with the nature of the substituent on position 1 of pyrazole, decreasing in the order pyridyl>Ph>>methyl. The activity for (Q(CF3,py))2SnCl2 on the SK-MEL-5 cell line is IC50 = 50 microM. PMID- 16313962 TI - DNA cleavage reaction induced by dimeric copper(II) complexes of N-substituted thiazole sulfonamides. AB - A new dinuclear copper(II) complex has been synthesised and structurally characterised: [Cu2(tz-ben)4] (Htz-ben = N-thiazol-2-yl-benzenesulfonamide). Its crystal structure, magnetic properties and electronic paramagnetic resonance (EPR) spectra were studied in detail. In the compound the metal centres are bridged by four non-linear triatomic NCN groups. The coordination geometry of the copper ions in the dinuclear entity is distorted square pyramidal (4+1). Two thiazole N and two sulfonamido N atoms occupy the equatorial positions and one sulfonamido O atom is in the axial position. Magnetic susceptibility data show a strong antiferromagnetic coupling, -2J = 114.1 cm(-1). The EPR spectra of a polycrystalline sample of compound has been obtained at the X- and Q-band frequencies at different temperatures. Above 20K the spectra are characteristic of S = 1 species with a zero field splitting parameter D = 0.4 cm(-1). The EPR parameters are discussed in terms of the known binuclear structures. The chemical nuclease ability of the title complex and that of the related [Cu2(tz-tol)4] compound (Htz-tol = N-thiazol-2-yl-toluenesulfonamide) is reported. The participation of hydroxyl radicals and a singlet oxygen-like entity in the DNA cleavage reaction has been deduced from the assays with radical oxygen scavengers. PMID- 16313963 TI - The prevalence of HCV antibody in South Australian prisoners. AB - OBJECTIVES: The study was aimed at identifying the hepatitis C virus (HCV) antibody status of prisoners incarcerated in South Australia in order to develop an HCV prevalence estimate for the whole prison system. METHODS: The health records of persons incarcerated within eight prisons (accommodating approximately 93% of the jurisdiction's adult incarcerated population) were audited for evidence of HCV infection, age, sex, Indigenous status (Australian Aboriginal or Torres Strait Islander) and date of entry to prison. These data were analysed using both univariate and multivariate techniques. RESULTS: Among 1347 prisoners (1254 males and 93 females), 30.2% were HCV-antibody positive. After excluding those with no history of testing, HCV-antibody prevalence rose to 41.3% (males 39.8%, females 66.1%). HCV-antibody positivity was significantly associated with age, sex and Indigenous status in both univariate and multivariate analyses. CONCLUSIONS: Consistent with the literature, the prevalence of HCV infection in the SA prison system appears to be extremely high. This study suggests that HCV prevention efforts in prison settings should be considered as an important priority. PMID- 16313964 TI - Emergence of disseminated infections due to nontuberculous mycobacteria in non HIV-infected patients, including immunocompetent and immunocompromised patients in a university hospital in Taiwan. AB - OBJECTIVES: Although disseminated nontuberculous mycobacteria (NTM) infection has been increasingly reported in patients with AIDS, this condition is still considered rare in non-HIV-infected patients. METHODS: We retrospectively reviewed the medical records of non-HIV-infected patients treated at National Taiwan University Hospital from January 1997 to December 2004 to search for epidemiological trends in disseminated NTM infection. Data analyzed included etiology, demographic characteristics, underlying disease, initial symptoms and signs, treatment, and outcomes. RESULTS: Disseminated NTM infections occurred in 15 non-HIV-infected patients during the study period. The mean age of these patients was 51 years (range 13-80). Among these patients, eight were males, and eight had various immunocompromised conditions. Fever was an initial presentation in 12 patients (80%). Diagnosis was made by positive blood (seven patients) or bone marrow culture (five patients) in the majority (73%) of patients. Infectious etiologies included Mycobacterium avium complex (MAC) in eight, rapidly growing mycobacteria (RGM) in four and Mycobacterium kansasii in three. The mean time from initial presentation to initiation of anti-NTM therapy was 130 days (ranged from 9 days to 17 months). Mortality was highest in patients with M. kansasii (100%), followed by RGM (25%) and MAC (12.5%). CONCLUSIONS: Although disseminated NTM infection is an emerging condition among patients without HIV infection, it is a condition which is associated with immune deficiency. A high level of clinical suspicion should be maintained for avoiding delayed diagnosis, especially in patients with underlying immunosuppression. Blood culture might play an important role in the early diagnosis of disseminated NTM disease. Early diagnosis can lead to appropriate diagnosis and reduce mortality. PMID- 16313965 TI - Immune reconstitution syndrome associated with parvovirus B19-induced pure red cell aplasia during highly active antiretroviral therapy. AB - Persistent parvovirus B19 (B19) manifested as chronic pure red cell aplasia (PRCA) has been described in HIV patients. Immune reconstitution syndrome (IRS) is an immune response to an occult opportunistic pathogen or tumor antigen, usually occurs after antiretroviral treatment. We describe the first case of IRS associated with B19-induced PRCA during highly active antiretroviral therapy. PMID- 16313966 TI - Transient clonal expansion of T-large granular lymphocytes during primary cytomegalovirus infection. AB - Clonal immunoglobulin and T-cell receptor gene rearrangements are useful in distinguishing reactive lymphoproliferations from neoplastic processes. Here, we report a case of transient clonal expansion of CD8+ CD57- T-large granular lymphocytes (T-LGL) during primary cytomegalovirus infection. This case underlines that clonal expansion of T-LGL could be a reactive phenomenon related to an acute infectious disease and is not specific for lymphoid malignancy. PMID- 16313967 TI - Hydrocephalus in herpes simplex type 2 meningitis. AB - A 34-year-old woman presented to hospital with symptoms of meningitis, later confirmed to be due to herpes simplex virus type 2. She developed hydrocephalus on day 2 of her admission. We describe the first case of hydrocephalus associated with herpes simplex type 2 meningitis in an adult. PMID- 16313968 TI - Opposite effects of short- and long-duration isolation on ultrasonic vocalization, startle and prepulse inhibition in rats. AB - Housing conditions change the sensorimotor gating and the emotional state of rats. The prepulse inhibition (PPI) is a reduction in the startle reflex to loud sounds when they are preceded by acoustic stimuli of low-intensity, and this test has been considered to be a useful measurement of the functioning of sensory gating in animals and man. Rats reared under conditions of isolation from the time of weaning, but not for 1 week at adult age, display clear deficits in prepulse inhibition and in sensorimotor gating. Ultrasound vocalizations (USVs) at 20-24kHz are the usual defensive responses of rats exposed to threatening conditions such as novel situations. The amount of emissions of ultrasound vocalizations at these frequencies depends on whether the aversive stimuli are presented either alone or in combination. Given this background we evaluated the prepulse inhibition and the emission of ultrasound vocalizations in response to novelty in rats isolated for 1 day or 2 weeks and compared the results to those in grouped rats. We also examined whether the anxiolytic agent midazolam (0.5 and 1.0mg/kg) could reverse the effects of isolation under the experimental conditions used. Rats isolated for 1 day showed a significant increase in the number and duration of USVs together with an enhancement in the startle response to loud sounds, which were antagonized in a dose-dependent manner by midazolam. On the other hand, 2-week isolation had the effect of reducing the number of USVs emitted at 20-24kHz without changing the startle response. The PPI was not changed by isolation, irrespective of the duration of isolation (1 day or 2 weeks). The results suggest that 1 day and 2 weeks of isolation have opposite effects on the emotional state of the animals. While short periods of isolation cause an anxiolytic-sensitive enhancement of the defensive responses, longer periods tend to reduce the defensive reaction of the animals to aversive stimuli. Based on these results, this work presents a novel method for induction of two different modes of defensive response, which are proposed to be mediated by separate neural substrates in rats. Also, isolation from 1 day to 2 weeks has no effect on the expression of prepulse inhibition and, by extension, on the functioning of the sensory gating. PMID- 16313969 TI - Psychometric delineation of the most discriminant symptoms of depressive mixed states. AB - Recent studies have shown that depressive mixed state (DMX), i.e., a major depressive episode (MDE) plus 3 or more intra-MDE hypomanic symptoms, is prevalent in bipolar-II disorder (BP-II) and not uncommon in major depressive disorder (MDD) outpatients. The main aim of the present analyses was to find the most discriminant intra-MDE hypomanic signs and symptoms predicting depressive mixed state. Consecutive 602 MDE outpatients (348 BP-II, 254 MDD) were interviewed with the Structured Clinical Interview for DSM-IV, the Hypomania Interview Guide, and the Family History Screen, by a senior psychiatrist in a private practice. Sensitivity (SE), specificity (SP), positive predictive value (PPV), and negative predictive value (NPV) of individual intra-MDE hypomanic signs and symptoms for depressive mixed state showed that irritability had a balanced combination of SE, SP, PPV, NPV, and that psychomotor agitation and greater talkativeness had the highest SP and PPV, but lower SE. By virtue of reducing misdiagnoses, SP and PPV are the most important of these tests for clinicians. On the basis of SP and PPV, our results suggest that, though not the most prevalent, irritability (50.3%) and psychomotor agitation (27.7%) represent the most discriminant features of depressive mixed state. Presence of these features should serve as a "red flag" to help clinicians probe more skillfully for the diagnosis of depressive mixed state. Beyond its diagnostic significance, the foregoing signs and symptoms are among the clinical features the FDA lists as presaging "suicidality" on antidepressant monotherapy. PMID- 16313970 TI - Molecular cloning and mRNA expression of duck invariant chain. AB - In the present study we identified a duck invariant chain (Ii) cDNA, named duck Ii-1, by RT-PCR and RACE. It was 1190 bp in length and contained a 669 bp open reading frame. An alternative transcript encoding a thyroglobulin (Tg)-containing form of Ii, named duck Ii-2, was also found in duck. The putative amino acid sequence of duck Ii-1 showed an 82% similarity to chicken Ii-1 and about 60% similarity to its mammalian homologues. The similarity of the Tg domain between duck and chicken Ii-2 was 96%, and about 70% between duck and mammalian Ii. The result of RT-PCR showed that Ii mRNA was extensively expressed in various tissues. High levels of both Ii-1 and Ii-2 mRNA were observed in the spleen and bursa of Fabricius. The predicted three-dimensional (3D) structures of duck Ii trimerization and Tg domain are similar to the corresponding regions of human Ii analyzed by comparative protein modeling. These findings indicate that the two isoforms of duck Ii, which strongly expressed in the major immune organs, share structural identity with human Ii. PMID- 16313971 TI - Comparison of amino acid compositions of peptides eluted from HLA-B27 molecules of healthy individuals and patients with ankylosing spondylitis. AB - HLA-B27 is a relative risk factor for ankylosing spondylitis (AS) and is present in about 10% in European populations but in 95% of AS patients. Various data suggest that the HLA-B27 molecule itself could be the strongest risk factor, but there is no explanation for this association. To define differential antigen presenting features of HLA-B27 in healthy individuals and AS patients, a question that cannot be addressed by biochemical studies on cell lines, the HLA-B27 protein was purified from peripheral blood lymphocytes of AS patients and healthy controls and pool sequencing of the bound peptides was performed. Results show that peptides are rich in proline (Pro) and the content of arginine (Arg) is much lower in comparison with sequences listed in the register of peptides eluted from cell cultures. Statistically significant differences were detected in frequencies of a subset of amino acids, predominantly at positions in the middle of the peptides. The frequency of Glu was increased and Gln was decreased in peptides from AS patients. Detailed analysis of purity of the immunoisolated HLA molecules excluded that the peptides might originate from any co-purified HLA molecules other than B27. We conclude that statistically significant increase in the Glu/Gln ratio of peptides from AS patients, consistent with increased deamidation in vivo, may account for differential antigenicity of HLA-B27 in patients. Source protein(s) of deamidated peptides remain unknown. PMID- 16313972 TI - Human milk antibodies with polysaccharide kinase activity. AB - It was shown for the first time that a small fraction of milk secretory IgA (sIgA) is tightly bound to oligosaccharides (oligoSACs) and polysaccharides (polySACs). The ability of sIgA to phosphorylate oligo- and polysaccharides was shown to be an intrinsic property of this antibody. In contrast to known kinases, sIgAs with polysaccharide kinase activity can transfer phosphoryl group to oligo- and polysaccharides not only from [gamma-(32)P]ATP but can also use [(32)P]orthophosphate as a substrate of phosphorylation reaction. An extremely unusual property of polysaccharide kinase Abs is their high affinity for orthophosphate (K(m) = 15-77 microM), and orthophosphate is a better substrate than ATP. Two first examples of natural abzymes (Abzs) with synthetic activity were milk sIgA with protein and lipid kinase activities. Polysaccharide kinase sIgA of human milk is the third example of natural antibodies (Abs) with synthetic activity. PMID- 16313973 TI - Enhancement of antitumor immunity of dendritic cells pulsed with heat-treated tumor lysate in murine pancreatic cancer. AB - Cancer vaccines using dendritic cells (DCs) have been shown to induce antitumor immunity and have recently been applied to non-immunogenic cancers, such as pancreatic cancer. In this study, we utilized DCs loaded with heat-treated tumor lysate (HTL-DC) as a vaccine in order to stimulate antitumor immunity in a murine pancreatic cancer model and compared them to DCs loaded with tumor lysate (TL DC). The poorly immunogenic mouse ductal pancreatic cancer cell line PANC02 with syngeneic mouse strain C57BL/6 was used as a model. Inducible heat shock proteins (HSPs) were significantly increased in HTL (HSP70 and HSP90). Tumor size measurements indicated that HTL-DC induced stronger tumor suppression than unpulsed DC or TL-DC (43% reduction in tumor volume compared to control group). T cell proliferation assay and IFN-gamma ELISPOT assay showed that T cell activation increased in the following order: DC100%) CQ accumulation in the FCM29 strain in a glucose-free medium while the addition of glucose further significantly increased this accumulation. Our study therefore clearly demonstrates that MG prevents CQ efflux from, and stimulates CQ influx into, drug-resistant Pf. Overall, MG appears to be a useful lead for the design and synthesis of more powerful and effective resistance modulators. PMID- 16313983 TI - Mitochondrial DNA haplogroups in Spanish patients with hypertrophic cardiomyopathy. AB - Mutations in mtDNA have been implicated in the development of hypertrophic cardiomyopathy (HCM), including cases from families with a maternal transmission. Alleles at several polymorphic sites in mtDNA define different haplogroups and some of these haplogroups have been involved in the risk of developing several diseases in which mitochondria should be involved. We analysed the association between the nine common European haplogroups and HCM. A total of 130 Spanish patients and 300 healthy controls were genotyped for eight mitochondrial single nucleotide polymorphisms (SNPs) through polymerase chain reaction followed by digestion with a restriction enzyme (PCR-RFLP). We compared the frequencies of these polymorphisms and mitochondrial haplogroups between patients and controls. Haplogroup T, specifically defined by 13368A, was significantly involved in the risk of developing HCM in our population (p=0.007; OR=2.42; 95% CI=1.25-4.67). Our data suggest that the genetic variation at the mitochondrial genome could significantly contribute to the risk for HCM. PMID- 16313984 TI - ABCA1 polymorphisms and prognosis after myocardial infarction in young patients. AB - High-density cholesterol (HDL) levels are affected by genetic influences and certain behaviors. Low levels of HDL-C are considered as an independent risk factor for premature coronary heart disease. In patients with Tangier disease, characterised by low HDL levels, mutations in the ATP binding cassette transporter have been described. We have analysed three polymorphisms of the ABCA1 gene (-477C/T, R219 K, and I883M) in a cohort of young male survivors of myocardial infarction in order to know their influence in long-term prognosis. In premature heart disease, knowing prognosis factors is specially relevant. PMID- 16313985 TI - Moray mouth heart. PMID- 16313986 TI - Nicotinic modulation of human auditory sensory memory: Evidence from mismatch negativity potentials. AB - Impairment in mismatch negativity (MMN) generation is a robust biological marker of schizophrenia. Understanding the physiological and pharmacological processes involved in its generation may therefore advance our understanding of this complex disorder. The present study tested if acute administration of nicotine modulates human auditory sensory memory as measured with MMN. ERP responses to tone duration deviants were recorded using a stimulation protocol with continuously changing (roving) standard stimuli in order to measure the effect of stimulus repetitions on encoding of new stimuli (MMN memory trace effect). Twenty healthy adult volunteers were randomly assigned to receive either a nicotine gum or placebo after a baseline ERP recording. Nicotine administration augmented MMN amplitude in the treatment group compared to the baseline recording, while no MMN change was found in the placebo group. The drug effect was due to a selective enhancement of a frontal positive potential to standard stimuli (from 80-200 ms post-stimulus), while the negativity to deviants remained unaffected. Furthermore, under nicotine stimulation this repetition positivity showed a more marked increase with stimulus repetition compared to baseline and placebo. These results have potential implications for schizophrenia by suggesting that nicotinic agonists could ameliorate patients' MMN deficits by improving stimulus encoding and sensory memory trace formation. PMID- 16313987 TI - Complex slow potential generators in a simplified attention paradigm. AB - We have recently obtained evidence for complex multifocal, individually variable generators of slow cortical potentials, elicited during performance of visual tasks involving expecting attention, comparison and memory [Basile, L.F.H., Ballester, G., Castro, C.C., and Gattaz, W.F., 2002. Multifocal slow potential generators revealed by high-resolution EEG and current density reconstruction. Int. J. Psychophysiol., 45 (3), 227-240; Basile, L.F.H, Baldo, M.V., Castro, C.C., and Gattaz, W.F. 2003. The generators of slow potentials obtained during verbal, pictorial and spatial tasks. Int. J. Psychophysiol., 48, 55-65]. The cue target aspect of traditional paradigms for attention studies is equivalent to 'warning S1'-'imperative S2' in slow potential designs. We simplified Posner's spatial cueing task [Posner, M.I. 1980. Orienting of attention.Q. J. Exp. Psychol. Feb;32 (1), 3-25; Posner, M.I., Snyder, C.R., Davidson, B.J. 1980. Attention and the detection of signals. J Exp Psychol. Jun; 109 (2), 160-174] to temporal cuing only, by using visual cues to indicate the mere presence, on a known central position, of the eventual target (17 ms duration, +/-0.3 degrees grey circle). We recorded slow potentials on 12 healthy subjects, by 124-channel EEG system (Neuroscan Inc.), and modeled their generators using current density reconstruction (CDR) by L(p) 1.2 norm minimization ("Curry V4.6", Neurosoft Inc.) applied to the target onset time. MRIs were obtained for each subject for constraining source models to individual brain anatomy. Average slow potentials were computed from above 60 artifact-free EEG-epochs (ISI=1.6 s, average ITI=2.5 s). We tabulated individual cortical current distributions by cytoarchitectonic area of Brodmann, after scaling into negligible, low, moderate and strong local density, based on percentile bands with respect to absolute maximum current. Despite the task's simplicity, the main result was individual variability and complexity in both scalp voltage and cortical current distributions. As observed in our previous studies, there was strong intersubject variability in the exact distribution of task-related cortical activity. Only parietal area 7 bilaterally was non-negligibly active in all subjects (currents above 10% maximum). As opposed to drawing conclusions based on group averaged data, we propose that activity by cytoarchitectonic area be ranked and statistically analysed only after being scaled on each individual. Based on the present results, the concept of a universal attention-related set of cortical areas if restricted to common areas across subjects is challenged, since even area 7 may no longer be common when the sample size becomes larger. We discuss the fact that group averaging may de-emphasize weakly but consistently active areas, and emphasize strongly but inconsistently active ones. PMID- 16313988 TI - Event-related potentials in people at risk for vascular dementia. AB - This study examined the relationship between the integrity of cerebrovascular microcirculation, neuropsychological testing and event-related potential indices of cognitive functioning in a nonclinical group of participants being at risk for vascular dementia. Sonographic measures, magnetic resonance (MR) scans and ERPs were recorded in 30 participants treated for arterial hypertension, with no report of neurological or psychiatric disorders. As a sonographic measure of cerebral microcirculation, the arteriorvenous cerebral transit time (cTT) was recorded. While neuropsychological measures of memory functions and general mental ability functions did not show systematic correlations with the cTT and other measures of vascular pathology, a pronounced correlation was obtained between P3a latency and cTT. Participants with long cTT showed a delayed P3a. These findings suggest that the P3a is a sensitive measure for reduced cognitive functions even at early stages of cerebrovascular pathology and by this may be a valuable tool for the early identification of cognitive deficits in individuals being at risk for vascular dementia. PMID- 16313989 TI - Differential responsiveness of c-Fos expression in the rat medulla oblongata to different treadmill running speeds. AB - Expression of the inducible transcription factor c-Fos was mapped in the rat medulla oblongata to identify the brain areas respond to different running speeds. Rats were subjected to 30 min of running, either at high speed, low speed or just sitting on a treadmill (control). Blood lactate levels were measured to confirm the physiological impact of different exercise intensities. The number of c-Fos-ir cells was counted and their spatial distributions were mapped through the rostral to the caudal level in the medulla. A statistically significant exercise intensity-dependent induction of c-Fos was observed in the nucleus of the solitary tract (NTS) and caudal ventrolateral medulla (CVL) in the medulla. Further, c-Fos induction was more predominant in the caudal part of each nucleus. The present data clearly show that different running speeds cause differential activation of each nucleus in the medulla, and in particular, the caudal parts of the NTS and the CVL are the most responsive to speed changes. The present study identifies brain areas newly found to be responsive to changes in running speed. These findings are likely to be particularly helpful in studies of specific neural circuits and their functions in response to different running speeds. PMID- 16313990 TI - Identification and analysis of the chivosazol biosynthetic gene cluster from the myxobacterial model strain Sorangium cellulosum So ce56. AB - Myxobacteria belonging to the genus Sorangium are known to produce a variety of biologically active secondary metabolites. Chivosazol is a macrocyclic antibiotic active against yeast, filamentous fungi and especially against mammalian cells. The compound specifically destroys the actin skeleton of eucaryotic cells and does not show activity against bacteria. Chivosazol contains an oxazole ring and a glycosidically bound 6-deoxyglucose (except for chivosazol F). In this paper we describe the biosynthetic gene cluster that directs chivosazol biosynthesis in the model strain Sorangium cellulosum So ce56. This biosynthetic gene cluster spans 92 kbp on the chromosome and contains four polyketide synthase genes and one hybrid polyketide synthase/nonribosomal peptide synthetase gene. An additional gene encoding a protein with similarity to different methyltransferases and presumably involved in post-polyketide modification was identified downstream of the core biosynthetic gene cluster. The chivosazol biosynthetic gene locus belongs to the recently identified and rapidly growing class of trans-acyltransferase polyketide synthases, which do not contain acyltransferase domains integrated into the multimodular megasynthetases. PMID- 16313991 TI - HearSNPV orf83 encodes a late, nonstructural protein with an active chitin binding domain. AB - The ORF83 (ha83) of Helicoverpa armigera nucleopolyhedrovirus (HearSNPV) was characterized during the present study. Sequence analysis and chitin-binding assay revealed that Ha83 contained an active chitin-binding domain. Northern blot and Western blot analyses demonstrated that ha83 was expressed as a late gene and encoded a nonstructural protein of HearSNPV. Ha83 gene was transcribed beginning at 12h post-infection in infected Helicoverpa zea cells (HzAM1). Western blot analysis using a rabbit derived polyclonal antibody showed the product of ha83 in infected cells was a 20 kDa protein, in tune with the theoretical size of 18.8 kDa. The protein was first detected in the cytoplasm of infected HzAM1 cells at 12h p.i., and was transported later into the nucleus during infection. PMID- 16313992 TI - Completion of the genome sequence of Lettuce necrotic yellows virus, type species of the genus Cytorhabdovirus. AB - We completed the genome sequence of Lettuce necrotic yellows virus (LNYV) by determining the nucleotide sequences of the 4a (putative phosphoprotein), 4b, M (matrix protein), G (glycoprotein) and L (polymerase) genes. The genome consists of 12,807 nucleotides and encodes six genes in the order 3' leader-N-4a(P)-4b-M-G L-5' trailer. Sequences were derived from clones of a cDNA library from LNYV genomic RNA and from fragments amplified using reverse transcription-polymerase chain reaction. The 4a protein has a low isoelectric point characteristic for rhabdovirus phosphoproteins. The 4b protein has significant sequence similarities with the movement proteins of capillo- and trichoviruses and may be involved in cell-to-cell movement. The putative G protein sequence contains a predicted 25 amino acids signal peptide and endopeptidase cleavage site, three predicted glycosylation sites and a putative transmembrane domain. The deduced L protein sequence shows similarities with the L proteins of other plant rhabdoviruses and contains polymerase module motifs characteristic for RNA-dependent RNA polymerases of negative-strand RNA viruses. Phylogenetic analysis of this motif among rhabdoviruses placed LNYV in a group with other sequenced cytorhabdoviruses, most closely related to Strawberry crinkle virus. PMID- 16313993 TI - Injectable porous hydroxyapatite microparticles as a new carrier for protein and lipophilic drugs. AB - Hydroxyapatite (Ca10 (PO4)6(OH)2) is a biodegradable material that forms a major component of bones and teeth. We prepared injectable spherical porous hydroxyapatite microparticles (SP-HAp) as a drug carrier by the spray-drying method. We then examined the usefulness of SP-HAp as a carrier for drugs such as interferon alpha (IFNalpha), testosterone enanthate (TE), and cyclosporin A (CyA). SP-HAp had an average diameter of 5 mum and a porosity of approximately 58%. It could be injected subcutaneously through a 27-gauge needle. SP-HAp was observed to be biodegradable. The speed of degradation of SP-HAp could be regulated by altering the calcination temperature. IFNalpha was adsorbed well to SP-HAp particles, but INFalpha was released faster from the particles, than the particles could degrade in both in vitro and in vivo experiments. Addition of human serum albumin and zinc (reinforcement) to IFNalpha-adsorbed SP-HAp caused marked prolongation of release in vivo. The in vivo release of testosterone enanthate and CyA from SP-HAp preparation, which was easily injectable, was similarly prolonged to that from the oil preparation. In conclusion, the SP-HAp seems to be useful as a biodegradable and subcutaneously injectable drug carrier. It is suggested that the reinforcement of the SP-HAp is very effective on the sustained release of drugs. PMID- 16313994 TI - Relationships between skin's electrical impedance and permeability in the presence of chemical enhancers. AB - Stratum corneum, the outermost layer of the skin, offers a strong barrier to the movement of solutes as well as ions. We report on the existence of a relationship between skin's electrical impedance and its permeability to hydrophilic (mannitol and inulin) as well as hydrophobic (corticosterone and estradiol) solutes in the presence of 33 distinct chemical penetration enhancer formulations. The correlation between impedance and permeability was excellent (r2=0.8) for hydrophilic solutes and moderate, yet significant (r2 approximately 0.5), for hydrophobic solutes. The possibility of using skin's electrical impedance to choose potent formulations was quantitatively assessed. Percentile ranking of penetration enhancers based on their effect on skin's electrical impedance matched well with the ranking based on their effect on solute permeability (r2>0.9 for both mannitol and estradiol). These studies demonstrate the feasibility of using skin's electrical impedance to screen potent chemical enhancers. PMID- 16313995 TI - Chronic high glucose inhibits albumin reabsorption by lysosomal alkalinization in cultured porcine proximal tubular epithelial cells (LLC-PK1). AB - Lysosomal acidification is a key step of albumin reabsorption in proximal tubular epithelial cells (PTECs). This study was performed to examine the influence of chronic high glucose on lysosomal acidification in cultured PTECs. Porcine PTECs (LLC-PK(1) cells) were cultured in 16.7 mM (300 mg/dl) glucose (HG) alone or with 0.5 mM phlorizin for 24 weeks and subsequently for 12 weeks in 5.5 mM (100 mg/dl) glucose (NG). Chronic HG inhibited the fluorescein isothiocyanate (FITC)-albumin (A) uptake progressively, while phlorizin reversed the inhibition. NG for 12 weeks after HG normalized the uptake. The time-dependent uptake of FITC-A was inhibited by HG and bafilomycin A(1) (BafA(1)) after 15 min and by 4,4' diisothiocyanato-2,2'-disulfonic acid (DIDS) and N-ethyl-N-isopropyl-amiloride (EIPA) after 3 min. Cellular ATP was depleted by HG and restored by NG. Lysosomal pH, assessed by an acidotropic fluorescent probe, was alkalinized (pH 4.5-7.8) with 5.5-27.8 mM glucose and normalized by subsequent NG. BafA(1) alkalinized lysosomes, and the concentration required to 50% change for the pH and 50% inhibition of FITC-A uptake was similar. EIPA inhibited FITC-A uptake, but did not influence lysosomal pH. DIDS inhibited FITC-A uptake, and unexpectedly lowered lysosomal pH. Real time PCR showed that HG reduced the mRNA level for vacuolar H(+)-ATPase, but did not alter those of chloride channel-5 and Na(+) H(+)-exchanger-3. In conclusion, the chronic HG inhibits albumin reabsorption by lysosomal alkalinization in PTECs, probably due to ATP depletion and down regulation of vacuolar H(+)-ATPase. PMID- 16313996 TI - Role of pH in protection by low sodium against hypoxic injury in isolated perfused rat livers. AB - BACKGROUND/AIMS: The purpose of the present study was to characterize the role of Na+, pH and cellular swelling in the pathogenesis of hypoxic injury to rat livers. METHODS AND RESULTS: When livers were perfused with hypoxic Krebs Henseleit bicarbonate buffer (KHB) containing 143 mM Na+, release of LDH began after 30 min and was maximal after 60 min. In livers perfused with choline substituted low-Na+ KHB (25 mM Na+), LDH release began after 60 min and peaked after 120 min or longer. Supplementation of KHB with mannitol, a permeant sugar with antioxidant properties, suppressed LDH release, whereas sucrose, an impermeant disaccharide, did not afford protection. At the end of hypoxic perfusions with KHB and low-Na+ KHB, liver weight was not different, whereas mannitol but not sucrose increased liver weight after hypoxia. At pH 7.4, monensin, a Na+-H+ ionophore, reversed protection against hypoxia by low-Na+ KHB (10 mM Na+) but had no effect at pH 6.8. As measured directly by confocal microscopy of biscarboxyethylcarboxyfluorescein fluorescence, pH was lower during perfusion with low-Na+ KHB than KHB. CONCLUSIONS: Cytoprotection by low Na+ was not mediated by prevention of Na+-dependent tissue swelling. Rather, promotion of intracellular acidification likely mediates cytoprotection in low-Na+ buffer. PMID- 16313997 TI - Specialist consultations in primary health care-a possible substitute for hospital care? AB - This study assesses how the use of hospitals is affected by GPs being given an opportunity to send their patients to private specialists for consultation instead of referring them exclusively to hospital outpatient clinics. In the City of Turku three populations were served for a period of over 34 months by different service models. The first model was implemented in municipal health centres by 10 GPs with a list system and an option to consult private specialists. In the second model there were four GPs in municipal health centres without a list system or the consultation option. The third model comprised four private GPs with a list system and the consultation option. Persons with private GPs with a list system had fewer visits to the hospital outpatient clinics and fewer bed days than persons served by a municipal GP, either with or without a list system. When GP's have the opportunity to send patients for consultation to private specialists, both the number of visits to hospital outpatient clinics and the number of bed days are reduced. PMID- 16313999 TI - Twenty-year secular changes in sex specific lung cancer incidence rates in an urban Chinese population. AB - The objective of this study was to describe trends in the incidence rates of primary lung cancer in a geographically defined Chinese population. Primary lung cancer cases (N=40,022) diagnosed between 1981 and 2000 were identified by the Tianjin Cancer Registry. Age-specific and age-adjusted incidence rates to the world standard population were examined in both males and females. Age-period cohort (APC) model and Poisson regression were used to assess the cohort effects and incidence trends. Crude and age-adjusted incidence rates in the study period were: 66.2/100,000 and 45.2/100,000 in males; and 47.7/100,000 and 28.2/100,000 in females, respectively. The major birth cohort effect can be described as for those born before 1940, in every age group lung cancer incidence rate increased as the birth years advanced. For those born after 1940, age specific incidence rates decreased as the birth years advanced. Results from the Poisson regression analyses suggested a statistically significant increasing trend of incidence rates of lung cancer from 1981 to 1990 and changed little afterwards. Through first 10 years of the study period between 1981 and 2000, lung cancer incidence rates increased in both males and females. While the study results suggest that the age-adjusted incidence rates may have reached their peak and may even decline, as the Chinese population ages, and smoking prevalence remains high, the number of new lung cancer cases will continue to increase and overall burden of lung cancer will remain high. PMID- 16314000 TI - EGFR mutation status and prognosis for gefitinib treatment in Japanese lung cancer. PMID- 16314001 TI - Bilateral effects of unilateral intrastriatal GDNF on locomotor-excited and nonlocomotor-related striatal neurons in aged F344 rats. AB - In order to determine its effects on locomotor-related striatal electrophysiology in aged rats, glial cell line-derived neurotrophic factor (GDNF) was infused (vehicle or 30mug) into the right striatum of 24-25-month-old Fischer 344 (F344) rats. Multi-wire electrode arrays were then chronically implanted in striatum bilaterally. Thirty days later, striatal electrophysiological activity was recorded during freely moving conditions. Individual neurons were classified as locomotor-excited if they exhibited significant increases in firing rates during locomotor bouts versus periods of nonmovement. GDNF produced a significant increase in overall firing rates in locomotor-excited striatal neurons. This effect was observed in both the infused and the contralateral striatum. GDNF also attenuated the bursting activity of nonlocomotor-related striatal neurons, an effect that was also present bilaterally. These results suggest that GDNF's antiparkinsonism effects are associated with increased excitability of motor related striatal neurons and diminished activity of neurons that do not exhibit explicit motor-related changes in activity. Such studies may aid in understanding the mechanism of potential therapies for movement disorders seen in aging and Parkinson's disease. PMID- 16314002 TI - Undergraduate nursing students' acquisition and retention of CPR knowledge and skills. AB - The ability to respond quickly and effectively to a cardiac arrest situation rests on nurses being competent in the emergency life-saving procedure of cardiopulmonary resuscitation (CPR). The aim of this study was to investigate the extent to which Irish nursing students acquire and retain CPR cognitive knowledge and psychomotor skills following CPR training. A quasi-experimental time series design was used. A pre-test, CPR training programme, post-test, and re-test were conducted. CPR knowledge was assessed by a multiple-choice assessment and psychomotor skills were assessed by observing CPR performance on a Resusci-Anne skill-meter manikin. The findings showed an acquisition in nurses' CPR knowledge and psychomotor performance following a 4h CPR training programme. Despite this, at no point in this study, did any nurse pass the CPR skills assessment. A deterioration in both CPR knowledge and skills was found 10 weeks following CPR training. However, students' knowledge and skills were improved over their pre training scores, which clearly indicated a positive retention in CPR cognitive knowledge and psychomotor skills. The study findings present strong evidence to support the critical role of CPR training in ensuring that nursing students progress to competent and confident responders in the event of a cardiac related emergency. PMID- 16314003 TI - Artificial nutrition at the bioethic cross-road between treatment and basic health care. PMID- 16314004 TI - Water, mineral waters and health. AB - The authors focus on water resources and the use of mineral waters in human nutrition, especially in the different stages of life, in physical activity and in the presence of some morbid conditions. Mineral water is characterized by its purity at source, its content in minerals, trace elements and other constituents, its conservation and its healing properties recognized by the Ministry of Health after clinical and pharmacological trials. Based on total salt content in grams after evaporation of 1l mineral water dried at 180 degrees C (dry residues), mineral waters can be classified as: waters with a very low mineral content, waters low in mineral content, waters with a medium mineral content, and strongly mineralized waters. Based on ion composition mineral waters can be classified as: bicarbonate waters, sulfate waters, sodium chloride or saltwater, sulfuric waters. Based on biological activity mineral waters can be classified as: diuretic waters, cathartic waters, waters with antiphlogistic properties. Instructions for use, doses, and current regulations are included. PMID- 16314005 TI - Modulation of hydrogen peroxide-induced DNA damage, MAPKs activation and cell death in PC12 by ergothioneine. AB - BACKGROUND & AIMS: Ergothioneine (EGT) is a natural occurring compound, synthesized by soil bacteria in fungal substrates, exhibiting antioxidant functions in many cell models. The aim of this study was to assess the effect of EGT in the prevention of H2O2-dependent cell death and oxidative damage on a model of neural cell derived from rat pheocromocytoma, the PC12. METHODS: The ability of EGT was tested by the 3 (4,5-dimethylthiazol-2-yl) 2, 5 diphenyltetrazolium bromide (MTT) assay and Comet assay. H2O2 insult was challenged with increasing concentration of antioxidant using two different incubation periods: 1 and 23 h of EGT pre-treatment followed by 23 and 1 h of H2O2, respectively, for both the MTT and the Comet assay data. CONCLUSION: The pre-treatment for 23 h with EGT, 250 microM and 1mM, followed by 1h of H2O2 incubation at the concentration of 250 and 500 microM, resulted in increased cell viability (P < 0.001) compared to the H2O2 cell batch. This correlated with a decrease in DNA damage as visualized by the Comet assay. Moreover, protein analysis reveals that in the presence of 250 microM of H2O2, EGT acted as a p38 MAPK and Akt specific inhibitor. EGT may play a protective role in rescuing cells from stress-induced apoptosis, likely by activating an intracellular antioxidant pathway involving p38 MAPK genes cascade. PMID- 16314006 TI - Bovine immune colostrum against 17 strains of diarrhea bacteria and in vitro and in vivo effects of its specific IgG. AB - Bovine colostral antibodies of cows immunized with a multivalent vaccine consisting of whole cells of 17 strains of pathogenic diarrhea bacteria were generated, and the specific IgG with high activities and titres directed against these pathogens was purified using an ammonium sulfate precipitation and verified by SDS-PAGE. We demonstrate that specific IgG has a strong activity of inhibiting in vitro growth and colonization in pathogens by agglutinating with bacteria and destroying cell walls. Normal IgG purified from non-immunized bovine colostrum is incapable of eliciting the same consequences as specific IgG. Specific IgG prevents enteroinvasive Escherichia coli/Salmonella typhi-induced diarrhea and may exert an effective protection by enhancing splenic NK cell activity, elevating IL-2 level and inhibiting excessive release of TNF-alpha in mice. Thus, the specific IgG from colostral antibodies of immunized bovine can provide effective protection or therapy for multibacteria-induced diarrhea. PMID- 16314007 TI - The efficacy of mouse-brain inactivated Nakayama strain Japanese encephalitis vaccine--results from 30 years experience in Taiwan. AB - An intensive mandatory vaccination program has been underway, combating Japanese encephalitis (JE) since 1968 in Taiwan. Long-term collection of immunization records has been developed from 1967 to 2000 in this study to retrospectively assess the efficacy of the mouse-brain inactivated Nakayama JE vaccine. The vaccine efficacy (VE) of completing at least two doses of the JE vaccine was 96.98%. Among 1 to 14-year-old children, the efficacy of completing 1, 2, and 3 doses of immunization was 85.59%, 91.07% and 98.51%, respectively. Furthermore, the long-term efficacy for a single dose vaccinated at least 25 years was 86.79%, and for 2 and 3 doses it was 88.10% and 95.54%, respectively. In contrast to previous studies that recommended at least two doses of JE vaccination to acquire necessary protection, the empirical results in this study indicated that even immunization with one single dose provides sufficient protection to the population. However, a single dose of JE vaccine might still be beneficial for some JE epidemic or endemic developing countries with limited resources for infectious disease control. PMID- 16314008 TI - Two serine protease inhibitors (serpins) that induce a bovine protective immune response against Rhipicephalus appendiculatus ticks. AB - We have previously undertaken preliminary characterization of two Rhipicephalus appendiculutus serine protease inhibitors (RAS-1 and -2) as anti-tick vaccine candidates. In this study, to clarify this hypothesis, we generated and further characterized recombinant RAS-1 and -2 (rRAS-1 and -2) and tested their potency as a cocktail anti-tick vaccine in cattle. RT-PCR analysis showed that RAS-1 and 2 mRNA transcripts are expressed during all life cycle stages of ticks, independent of sex. As judged by SDS-PAGE rRAS-1 and -2 migrated as a molecular weight of around 64 and 60 kDa protein, respectively, considering that the expression vector produced a recombinant protein fused with 18-22 kDa TRX protein. RAS-1 and -2 were found not to be secreted into the bite site as determined by the reactivity of anti-tick saliva sera to rRAS-1 and -2, suggesting that both proteins are concealed antigens. Vaccination of cattle with a combination of rRAS-1 and -2 conferred significant protective immunity against ticks, resulting in 61.4% reduction in nymph engorgement rate, and in 28 and 43% increased mortality rate in adult female and male ticks, respectively. This is the first report on an anti-tick vaccine trial using a combination of two different serpins derived from R. appendiculatus, and using cattle as a natural host. PMID- 16314009 TI - Protective efficacy induced by Mycobacterium bovis bacille Calmette-Guerin can be augmented in an antigen independent manner by use of non-coding plasmid DNA. AB - Tuberculosis caused by infection with Mycobacterium tuberculosis or M. bovis remains one of the most important infectious diseases of man and animals, and continues to inflict a huge cost in both health and financial terms. The current vaccine, BCG demonstrates variable efficacy and so a more robust vaccine strategy to either replace or supplement BCG is required. We have utilised a DNA prime-BCG boost strategy in a murine M. bovis challenge model using a cocktail of 3 DNA vaccines (encoding Hsp65, Hsp70 and Apa) followed by BCG. Controls were inoculated with vector DNA only, coding DNA only, BCG only or vector DNA followed by BCG boost. Analysis of immune responses by ELISpot prior to challenge, revealed that the coding DNA/BCG prime boost resulted in an increased frequency of antigen-specific IFNgamma producing cells compared to the other regimes. When spleen cell cytokine production to BCG antigens was analysed, significantly more IFNgamma and IL-12 was seen in those groups primed with DNA (coding or vector) prior to BCG than those receiving BCG alone. Analysis of bacterial counts revealed that DNA priming followed by BCG boost further improved the protective immunity induced by BCG alone. Surprisingly, inoculation with vector DNA was as efficacious as the coding DNA in enhancing BCG protection. Taken together these results indicate that whilst the coding DNA vaccines induce antigen specific responses, treatment with the vector DNA is sufficient for the increase in protective immunity over that induced by BCG, suggesting that the vector DNA may be acting as a non-specific adjuvant for BCG immunization. PMID- 16314010 TI - Fish soluble Toll-like receptor (TLR)5 amplifies human TLR5 response via physical binding to flagellin. AB - Fish has a soluble form of TLR5 ortholog (TLR5S), which does not exist in mammals. We identified TLR5S from rainbow trout and named rtTLR5S, which was about 38% homologous to the extracellular domains of human (hu) and mouse TLR5. Adjuvancy of rtTLR5S to flagellin response by human TLR5 (huTLR5) was tested in this study. A chimera constructed of rtTLR5S and the intracellular TIR of huTLR5 expressed on HeLa cells signaled the presence of various species of bacterial flagellin resulting in NK-kappaB activation. huTLR5S, when co-expressed with rtTLR5S in HeLa cells, augmented response to flagellin resulting in robust huTLR5 mediated NF-kappaB activation. Physical binding of flagellin to rtTLR5S was detected under the conditions where huTLR5 induced rtTLR5S-amplified NF-kappaB activation. Signal amplification by rtTLR5S was specific to huTLR5: no other huTLRs tested were responded to rtTLR5S. These results suggest that the soluble TLR5 serves as an adjuvant augmenting flagellin-TLR5-mediated NF-kappaB activation even in human. PMID- 16314011 TI - The anatomy of the dorsal cutaneous branch of the ulnar nerve - a safe zone for positioning of the 6R portal in wrist arthroscopy. AB - The dorsal branch of the ulnar nerve passes close to the 6 Radial portal used in wrist arthroscopy. We examined 20 cadaveric limbs to establish the course of this nerve. We found it consistently crossed a line between the ulnar styloid and the fourth web space at an average of 2.4 cm from the ulnar styloid (range 1.8-2.8 cm). This represented 23% (+/- 2.5 STD range 19-27%) of the distance from the styloid process to the fourth web space. Recommendations are made to help avoid iatrogenic injury to the nerve during arthroscopy. The literature on the anatomy of this nerve and its variations and the clinical relevance of this knowledge are discussed. PMID- 16314012 TI - Impact of short-term and long-term elevated CO2 on emission of carbonyls from adult Quercus petraea and Carpinus betulus trees. AB - The study aimed to elucidate the effect of elevated CO2 in short- and long-term experiments on the emission potential of carbonyls from mature oak (Quercus petraea) and hornbeam (Carpinus betulus). The trees were investigated using a canopy crane established in a mixed forest in Switzerland. Short-term CO2 experiments were performed on single leaves with a gas exchange measuring unit by increasing CO2 in the enclosure before and during the measurements. Long-term CO2 fumigation was performed for one vegetation period by a webFACE design. Short term as well as long-term exposure of leaves resulted in no significant changes in carbonyl exchange in neither Q. petraea nor C. betulus. Carbonyl emission was not affected by stomatal closure under elevated CO2 levels. In contrast to the emission of isoprenoids from vegetation which is thought to be reduced at elevated atmospheric CO2, the exchange of carbonyls seems to be generally unaffected under these conditions. PMID- 16314013 TI - Influence of vegetation in mitigation of methyl parathion runoff. AB - A pesticide runoff event was simulated on two 10 m x 50 m constructed wetlands (one non-vegetated, one vegetated) to evaluate the fate of methyl parathion (MeP) (Penncap-M). Water, sediment, and plant samples were collected at five sites downstream of the inflow for 120 d. Semi-permeable membrane devices (SPMDs) were deployed at each wetland outflow to determine exiting pesticide load. MeP was detected in water at all locations of the non-vegetated wetland (50 m), 30 min post-exposure. MeP was detected 20 m from the vegetated wetland inflow 30 min post-exposure, while after 10d it was detected only at 10 m. MeP was measured only in SPMDs deployed in non-vegetated wetland cells, suggesting detectable levels were not present near the vegetated wetland outflow. Furthermore, mass balance calculations indicated vegetated wetlands were more effective in reducing aqueous loadings of MeP introduced into the wetland systems. This demonstrates the importance of vegetation as sorption sites for pesticides in constructed wetlands. PMID- 16314014 TI - Death, depression and 'defensive expansion': closing down smoking as an issue for discussion in GP consultations. AB - This paper examines routine primary care consultations in the UK where smoking is discussed using data from a larger study of the factors influencing discussion of smoking between general practitioners (GPs) and patients. In this study, consultations have been analysed with a focus on the termination of discussion about smoking, using an approach that is informed by the conversation analytic (CA) literature on professional/client interaction. In interviews from the previous larger study, GPs suggested two main reasons for not pursuing discussion of smoking in consultations. One reason was an overarching fear of damaging the GP/patient relationship. The second reason related to clinical judgement, where it was feared that an attempt to stop smoking might exacerbate a patient's existing condition, particularly their mental health. This paper suggests that, while this latter scenario of clinical judgement is borne out by the consultation data, there are two more subtle patient behaviours which are associated with GPs abandoning further discussion of smoking: patients' 'troubles telling', where the issue of smoking is de-emphasised in the face of other 'troubles', and 'defensive expansion', where the patient over-emphasises deficiencies to curtail discussion. Greater awareness of the situations in which doctors end discussion of smoking will help GPs to develop ideas for alternative approaches in these circumstances which could result in more meaningful, effective engagement between doctors and their patients who smoke when smoking is discussed. PMID- 16314015 TI - What factors induce health care decision-makers to use clinical guidelines? Evidence from provincial health ministries, regional health authorities and hospitals in Canada. AB - This paper addresses three questions: What is the extent of clinical guideline utilization by decision-makers in provincial health ministries, regional health authorities and hospitals in Canada? Are there differences between these work settings in regard to the extent of clinical guideline utilization? What are the determinants of clinical guidelines utilization in health ministries, regional health authorities and hospitals? Based on a survey of 899 decision-makers from Canadian provincial health ministries, regional health authorities and hospitals, the results indicate that there are large differences between work settings in regard to clinical guideline utilization. Not surprisingly, work settings like hospitals rely more intensively on clinical guidelines than the other work settings (health ministries or agencies and regional health authorities). The results of the regression models indicate that cognitive factors, social factors, technological factors, organizational factors and individual attributes significantly predict the utilization of clinical practice guidelines by decision makers. However, the results of the regression models also indicate that some factors that predict clinical guideline utilization by decision-makers in hospitals do not predict clinical guidelines utilization by decision-makers working in ministries or in regional health authorities. Therefore, these results suggest that customized interventions would be appropriate in order to efficiently increase the utilization of clinical practice guidelines in different work settings. The paper concludes with suggestions for future research. PMID- 16314016 TI - Systems of medicine and nationalist discourse in India: towards "new horizons" in medical anthropology and history. AB - While accepting medical "pluralism" as a historical reality, as an intrinsic value inherent in any medical system, and as an ideal or desired goal that any multicultural society ought to achieve, this paper argues the need to go beyond the liberal pluralist tendencies that have dominated the debate so far. It holds that while documenting or dealing with the "co-existence" of varied medical traditions and practices, we must not ignore or underplay issues of power, domination and hegemony and must locate our work in a larger historical, social and political context. With this perspective, and based essentially on Assembly proceedings, private papers, official documents and archival materials from the first half of the 20th-century, this paper identifies three major streams in the nationalist discourse in India: conformity, defiance and the quest for an alternative. It shows that while the elements of conformity to biomedicine and its dominance remained more pronounced and emphatic, those of defiance were conversely weak and at times even apologetic. The quest for alternatives, on the other hand, although powerful and able to build trenchant civilizational and institutional critique of modern science and medicine, could never find adequate space in the national agenda for social change. The paper further holds that although the "cultural authority" and hegemony of biomedicine over indigenous science and knowledge were initiated by the colonial state, they were extended by the mainstream national leaderships and national governments with far more extensive and profound implications and less resistance. In light of the growing global networking of "traditional", "complementary" and "alternative" health systems on the one hand and the hegemonic and homogenizing role and presence of multilateral organizations (such as the World Bank and IMF) in shaping national health policies on the other, such insights from history become extraordinarily important. PMID- 16314017 TI - Pure misallocation of "0" in number transcoding: a new symptom of right cerebral dysfunction. AB - To account for the mechanism of number transcoding, many authors have proposed various models, for example, semantic-abstract model, lexical-semantic model, triple-code model, and so on. However, almost all of them are based on the symptoms of patients with left cerebral damage. Previously, I reported two Japanese patients with right posterior cerebral infarction showing pure misallocation of "0" (omission: "40,265"-->"4,265," addition: "107"-->"1,007," transposition: "4,072"-->"4,702") both in writing and oral reading of Arabic numerals. To examine whether the pure misallocation of "0" is commonly observed in patients with right cerebral damage, I investigated writing and oral reading of Arabic numerals in 18 patients with right cerebral damage and 16 healthy controls. All patients with right cerebral damage showed pure misallocation of "0" both in writing and reading. The pure misallocation of "0" due to right cerebral damage cannot be explained by current models. It may be more useful to explain the phenomenon by regarding an Arabic numeral as graph on a two dimensional plane composed of two axes (place-holding values and digits). PMID- 16314018 TI - Parameterization of movement execution in children with developmental coordination disorder. AB - The Rhythmic Movement Test (RMT) evaluates temporal and amplitude parameterization and fluency of movement execution in a series of rhythmic arm movements under different sensory conditions. The RMT was used in combination with a jumping and a drawing task, to evaluate 36 children with Developmental Coordination Disorder (DCD) and a matched control group. RMT errors in space and in time were significantly larger for children with DCD. Omission of sensory information decreased the accuracy of movement parameterization in children with DCD more than in the control group, suggesting that children with DCD have more problems in building up an internal representation of the movement. Errors in time correlated significantly with the jumping and drawing task, while errors in space did not. Deficits of temporal movement parameterization might be one of the underlying causes of poor motor performance in some children with DCD. PMID- 16314019 TI - Effects of sleep deprivation on Color-Word, Emotional, and Specific Stroop interference and on self-reported anxiety. AB - The aim of this study was principally to assess the impact of sleep deprivation on interference performance in short Stroop tasks (Color-Word, Emotional, and Specific) and on subjective anxiety. Subjective sleepiness and performance on a psychomotor sustained attention task were also investigated to validate our protocol of sleep deprivation. Twelve healthy young subjects were tested at four hourly intervals through a 36-h period of wakefulness under a constant routine protocol. Analyses of variance for repeated measurements revealed that self assessment of sleepiness on a visual analogue scale as well as mean reaction time performance on the sustained attention task, both for the first minute and for 10 min of testing, were worsened by sleep deprivation. Analyses revealed an increase in self-reported anxiety scores on the STAI questionnaire but did not reveal any significant effect after sleep deprivation either on indexes of interference or on accuracy in Stroop tasks. However, analyses showed sensitivity to circadian effect on verbal reaction times in the threat-related (Emotional) and sleep related (Specific) Stroop tasks. We concluded that 36 h of prolonged wakefulness affect self-reported anxiety and Emotional Stroop task resulting in a cognitive slowing. Moreover, total sleep deprivation does not affect interference control in any of the three short Stroop tasks. PMID- 16314020 TI - Combined treatment of olanzapine and mirtazapine in anorexia nervosa associated with major depression. AB - Several studies have shown that olanzapine is effective in weight restoration and maintenance for patients with anorexia nervosa (AN). However, major depression is a very common comorbid psychiatric disorder associated with AN. Additional antidepressant therapy may be required for treating anorexic patients with major depression. The authors present a case of AN associated with major depression, who responded well to the combination treatment of olanzapine and mirtazapine. A 27-year-old Taiwanese woman was admitted because of severe weight loss, poor nutrition, amenorrhea, major depression, and starvation complications including hematological dyscrasis, electrolytes and endocrine imbalance, and sinus bradycardia. In additional to nutritional and medical treatments, the patient was given olanzapine 10 mg/day and mirtazapine 30 mg/day. She took the combined medications for six months. Meanwhile she received cognitive behavior therapy and family therapy. With these treatments, the patient's depression was in remission, her body weight was increased from 24 to 38 kg, and her body mass index was increased from 9.8 to 15.5. Our case suggests that the combined treatment of olanzapine and mirtazapine can be used in the treatment of AN associated with major depression. PMID- 16314021 TI - Microcystin-LR induced cellular effects in mammalian and fish primary hepatocyte cultures and cell lines: a comparative study. AB - The impact of microcystin-LR, one of the most common cyanobacterial toxins, on liver and gut cells originating from mammals and fish was compared. Upon exposure of human and rainbow trout (Oncorhynchus mykiss) cell lines up to 2.5 microM microcystin-LR, no alteration in cell viability was observed as assessed with three fluorescent indicators dyes, CFDA-AM, Alamar Blue and neutral red. The lack of sensitivity of the trout cell lines coincided with an absence of detectable mRNA levels of organic anion transporter polypeptide (OATP), which is implicated in the uptake of microcystin-LR. In contrast to the cell lines, primary rainbow trout and mouse hepatocytes showed damage to subcellular structures, particularly the lysosomes, as indicated by neutral red. This led us to propose a thus far undetected role of lysosomes as targets and mediators of microcystin-LR elicited cellular damage. An inhibitor of OATP, rifampicin, partly protected hepatocytes from this damage. Yet, the sensitivity of rainbow trout hepatocytes rapidly declined in culture, accompanied by decreasing levels of OATP mRNA. The sensitivity of mouse hepatocytes toward microcystin-LR also declined in culture but overall was about 25-fold greater than that of the trout cells. These differences mirror those observed in vivo and suggest the use hepatocytes for deciphering the species differences. PMID- 16314022 TI - Chemokine regulation in response to beryllium exposure in human peripheral blood mononuclear and dendritic cells. AB - Exposure to beryllium (Be) induces a delayed-type hypersensitivity immune reaction in the lungs of susceptible individuals, which leads to the onset of Be sensitivity and Chronic Beryllium Disease (CBD). Although some mechanistic aspects of CBD have begun to be characterized, very little is known about the molecular mechanisms by which Be activates the host immune response. To gain insight into the cellular response to Be exposure, we have performed global microarray analysis using a mixture of peripheral blood mononuclear and dendritic cells (PBMC/DCs) from a non-CBD source to identify genes that are specifically upregulated in response to BeSO(4) stimulation, compared to a control metal salt, Al(2)(SO(4))(3). We identified a number of upregulated immunomodulatory genes, including several chemokines in the MIP-1 and GRO families. Using PBMC/DCs from three different donors, we demonstrate that BeSO(4) stimulation generally exhibits an increased rate of both chemokine mRNA transcription and release compared to Al(2)(SO(4))(3) exposure, although variations among the individual donors do exist. We show that MIP-1 alpha and MIP-1 beta neutralizing antibodies can partially inhibit the ability of BeSO(4) to stimulate cell migration of PBMC/DCs in vitro. Finally, incubation of PBMC/DCs with BeSO(4) altered the binding of the transcription factor RUNX to the MIP-1 alpha promoter consensus sequence, indicating that Be can regulate chemokine gene activation. Taken together, these results suggest a model in which Be stimulation of PBMC/DCs can modulate the expression and release of different chemokines, leading to the migration of lymphocytes to the lung and the formation of a localized environment for development of Be disease in susceptible individuals. PMID- 16314023 TI - A clinical evaluation of posterior composite restorations: 17-year findings. AB - OBJECTIVE: Since an increasing number of composite restorations in posterior teeth are placed as a routine, this study was conducted to evaluate long-term survival of these restorations placed in general practice. METHODS: Patients from a private dental office that received restorations in posterior teeth between 1987 and 1988 with P-50 (3M) or Herculite XR (Kerr) resin composites were selected and invited to participate. Restorations were placed under rubber dam isolation. Dentine walls were covered with glass ionomer cement, and composites were placed according to manufacturer's instructions. Thirty-eight patients agreed to participate and signed an informed consent prior to the evaluation. Two calibrated operators worked independently in the evaluation, using modified USPHS criteria. Survival of restorations or subsets of restorations grouped on the basis of variables (material, tooth, cavity type and size) was determined using Kaplan-Meier survival curves. RESULTS: Ninety-eight failures were recorded among the 282 restorations providing a crude estimate of 34.8% failures. The survival rate was not significant for material (p=0.92) but was significant between tooth (lower premolars and lower molars, p<0.0001), cavity type (p<0.001) and size (p<0.001). The majority of restorations exhibited A or B scores for the evaluated criteria. The main failure cause was fracture of both composites. CONCLUSIONS: The clinical performance of posterior resin composite restorations evaluated was acceptable after 17-year evaluation. However, the probability of failure of resin composite restorations in molars, Class II, and large restorations is higher. PMID- 16314024 TI - Sulfated glycosaminoglycans of the periurethral tissue in women with and without stress urinary incontinence, according to genital prolapse stage. AB - OBJECTIVE: The objective was to determine sulfated glycosaminoglycans (GAG) of the extracellular matrix (ECM) in women with and without stress urinary incontinence according to genital prolapse stage. STUDY DESIGN: Periurethral tissue was obtained from 30 women who underwent surgery for urinary incontinence, for pelvic organ prolapse, or for other benign gynecologic conditions. Biopsy specimens were assessed by biochemical methods to characterize and quantify sulfated glycosaminoglycans. Measurements were made of total glycosaminoglycans, chondroitin sulfate, dermatan sulfate, and of heparan sulfate. Data were compared using the t-test. RESULTS: In two groups, dermatan sulfate was the most predominant glycosaminoglycan. Women with stress urinary incontinence had significantly more total sulfated glycosaminoglycans (p<0.05) and dermatan sulfate (p<0.05) than women without stress urinary incontinence. We did not observe any differences in chondroitin sulfate and heparan sulfate. CONCLUSIONS: Women with stress urinary incontinence showed quantitative and qualitative differences in the biochemical characteristics of the extracellular matrix in periurethral tissue by analysis of sulfated glycosaminoglycans, according to genital prolapse stage. PMID- 16314025 TI - The value of cervical length and plasma proMMP-9 levels for the prediction of preterm delivery in pregnant women presenting with threatened preterm labor. AB - OBJECTIVE: To assess the value of plasma proMMP-9 levels in predicting the progress of threatened preterm labor to true preterm labor and delivery within 7 days of presentation, along with the ultrasonographic measurement of cervical length. STUDY DESIGN: The study included sixty two pregnant women presented in the 2nd Department of Obstetrics and Gynecology of University of Athens at Aretaieion Hospital with threatened preterm labor. On subjects' presentation, cervical length was evaluated with transvaginal ultrasonography, plasma proMMP-9 levels were measured with the use of a solid phase enzyme immunoassay, and maternal characteristics (smoking, history of preterm delivery and parity) as well as the use of tocolytics were recorded. Subjects were followed up for premature delivery within 7 days. ROC analysis for plasma proMMP-9 levels was performed and logistic regression analysis was used for the evaluation of the prognostic effect of the parameters. RESULTS: The only significant predictors for establishment of true preterm labor and delivery within 7 days of presentation were cervical length (with cutoff value 15 mm: odds ratio=0.022, 95% CI=0.002 0.243, p=0.002) and plasma proMMP-9 (with cutoff value 67.15 ng/ml: odds ratio=0.035, 95% CI=0.003-0.388, p=0.006). The combination of the adopted proMMP 9 and cervical length cutoff values was characterized by a sensitivity of 90.9% and a specificity of 98.3% for predicting the progress to true preterm labor and delivery. CONCLUSION: Plasma proMMP-9 levels could possibly serve as a predictive factor for the progress of threatened preterm labor to true preterm labor and delivery within 7 days of presentation, along with the ultrasonographic evaluation of cervical length. PMID- 16314026 TI - Rate of iron transfer through the horse spleen ferritin shell determined by the rate of formation of Prussian Blue and Fe-desferrioxamine within the ferritin cavity. AB - Iron (2+ and 3+) is believed to transfer through the three-fold channels in the ferritin shell during iron deposition and release in animal ferritins. However, the rate of iron transit in and out through these channels has not been reported. The recent synthesis of [Fe(CN)6]3-, Prussian Blue (PB) and desferrioxamine (DES) all trapped within the horse spleen ferritin (HoSF) interior makes these measurements feasible. We report the rate of Fe2+ penetrating into the ferritin interior by adding external Fe2+ to [Fe(CN)6]3- encapsulated in the HoSF interior and measuring the rate of formation of the resulting encapsulated PB. The rate at which Fe2+ reacts with [Fe(CN)6]3- in the HoSF interior is much slower than the formation of free PB in solution and is proceeded by a lag period. We assume this lag period and the difference in rate represent the transfer of Fe2+ through the HoSF protein shell. The calculated diffusion coefficient, D approximately 5.8x10( 20) m2/s corresponds to the measured lag time of 10-20 s before PB forms within the HoSF interior. The activation energy for Fe2+ transfer from the outside solution through the protein shell was determined to be 52.9 kJ/mol by conducting the reactions at 10 approximately 40 degrees C. The reaction of Fe3+ with encapsulated [Fe(CN)6]4- also readily forms PB in the HoSF interior, but the rate is faster than the corresponding Fe2+ reaction. The rate for Fe3+ transfer through the ferritin shell was confirmed by measuring the rate of the formation of Fe-DES inside HoSF and an activation energy of 58.4 kJ/mol was determined. An attempt was made to determine the rate of iron (2+ and 3+) transit out from the ferritin interior by adding excess bipyridine or DES to PB trapped within the HoSF interior. However, the reactions are slow and occur at almost identical rates for free and HoSF-encapsulated PB, indicating that the transfer of iron from the interior through the protein shell is faster than the rate-limiting step of PB dissociation. The method described in this work presents a novel way of determining the rate of transfer of iron and possibly other small molecules through the ferritin shell. PMID- 16314027 TI - Thermodynamically based profiling of drug metabolism and drug-drug metabolic interactions: a case study of acetaminophen and ethanol toxic interaction. AB - Drug-drug metabolic interactions can result in unwanted side effects, including reduced drug efficacy and formation of toxic metabolic intermediates. In this work, thermodynamic constraints on non-equilibrium metabolite concentrations are used to reveal the biochemical interactions between the metabolic pathways of ethanol and acetaminophen (N-acetyl-p-aminophenol), two drugs known to interact unfavorably. It is known that many reactions of these pathways are coupled to the central energy metabolic reactions through a number of metabolites and the cellular redox potential. Based on these observations, a metabolic network model has been constructed and a database of thermodynamic properties for all participating metabolites and reactions has been compiled. Constraint-based computational analysis of the feasible metabolite concentrations reveals that the non-toxic pathways for APAP metabolism and the pathway for detoxifying N-acetyl-p benzoquinoneimine (NAPQI) are inhibited by network interactions with ethanol metabolism. These results point to the potential utility of thermodynamically based profiling of metabolic network interactions in screening of drug candidates and analysis of potential toxicity. PMID- 16314028 TI - Interview with: Claude Schulman. PMID- 16314029 TI - Drug treatment for LUTS and BPH: new is not always better. PMID- 16314030 TI - Urothelial differentiation in vesicoureteric reflux and other urological disorders of childhood: a comparative study. AB - OBJECTIVE: The strong familial basis of vesicoureteric reflux (VUR) is well recognised, however little progress has been made in identifying the causative genes. In this study we have investigated whether human vesicoureteric reflux (VUR) is associated with the aberrant expression of urothelial differentiation associated antigens in view of the VUR phenotype of the Uroplakin IIIa (UPIIIa) "knockout" mouse. METHODS: Samples of urothelium were collected from 13 children with primary VUR, four children with secondary VUR and from seven children with non-refluxing disorders of the urinary tract. Immunohistochemistry was used to assess the expression of five uroplakin (UP) and cytokeratin (CK) differentiation restricted antigens, UPIa, UPIb, UPIIIa, CK13 and CK20. Samples were ranked blind according to immunohistochemical patterns relating to the differentiation associated distributions of the five antigens and the results were analysed statistically using the Kruskal-Wallis test. RESULTS: No objective differences in urothelial morphology or the expression of the five differentiation antigens were discernable in the urothelium of children with primary VUR, when compared with urothelium of children with a range of other pathology including VUR associated with duplication or pelvic renal ectopia, VUR secondary to outflow obstruction and non-refluxing upper tract obstruction. The p-values ranged from 0.168-0.651 and were not considered statistically significant. CONCLUSION: The results indicate that primary VUR is not associated with any major, collective abnormality of urothelial differentiation in man. In particular our findings provide no support for the suggestion that abnormalities of UPIIIa expression are implicated in the aetiology of human primary VUR. PMID- 16314031 TI - Nerve sparing endoscopic extraperitoneal radical prostatectomy--effect of puboprostatic ligament preservation on early continence and positive margins. AB - INTRODUCTION: A technical modification of nerve sparing endoscopic extraperitoneal radical prostatectomy (nsEERPE) with preservation of the puboprostatic ligaments is presented and compared to a previous technique. MATERIALS AND METHODS: nsEERPE was performed in 100 men with clinically localized adenocarcinoma of the prostate from March 2004 through February 2005. Patients were divided into two groups: Group A included patients in whom a standard nsEERPE was performed (n=50), and group B included patients in whom a puboprostatic ligament sparing nsEERPE was performed (n=50). The postoperative follow-up was 2 weeks and 3 months evaluating preliminary effects on early continence and positive margins. RESULTS: The early return to continence at 2 weeks postoperatively was achieved by 6 patients (12%) in group A, and 12 patients (24%) in group B. Three months after the procedure 24 (48%) and 38 (76%) patients were continent, in groups A and B respectively. Clinical outcome (early continence) was significantly better for Group B patients, at 2 weeks (chi-square test, p=0.0019) and at 3 months (chi-square test, p=0.0347) following surgery. No cases of complete or severe incontinence (more than 5 pads/day) were observed at 3 months after surgery, in either groups. Groups A and B did not exhibit significant differences regarding their histological status. In group A, positive margins were detected in 6.5% and 26.3% of patients with pT2 and pT3, respectively. In group B, positive margins were found in 3.2% and 15.8% of patients with pT2 and pT3, respectively CONCLUSION: We propose the use of puboprostatic ligament-sparing nsEERPE as an intriguing method to ascertain recuperation of early continence after nerve sparing procedures, without hindering the final oncological outcome. PMID- 16314032 TI - Value of counting white blood cells (WBC) in semen samples to predict the presence of bacteria. AB - OBJECTIVES: To investigate the correlation between the presence of white blood cells (WBC) without the use of specific stain to differentiate leukocytes and the presence of bacteria in semen samples of infertile men. METHODS: A total of 143 semen samples of men who attended an andrologic clinic for the evaluation of fertility were investigated using routine semen analysis (according to WHO laboratory guidelines) and bacterial culture. RESULTS: WBC were found in 43.4% (62/143). There were no WBC in 56.6% (81/143) of the samples (group I) while WBC were found in 43.4% (62/143) of the samples (group II). Pathogenic bacteria were detected in 48.2% (39/81) in group I and in 54.9% (34/62) in group II, all in all Bacteriospermia was present in 51.1% (73/143). The most common bacteria were Ureaplasma urealyticum, Enterococcus faecalis and Escherichia coli (23.8%, 16.8%, and 7.0% of samples, respectively). The sensitivity/specificity for detecting bacteria was 0.47/0.60 at a cut-off level of 0.25 Mio/mL WBC and 0.16/0.84 at a cut-off level of WBC 1 Mio/mL, representing likelihood ratios of 1.16 and 1.04, respectively. The greatest ratio between sensitivity and specificity (0.37/0.72) was found at a cut-off level of 0.5 Mio/mL WBC, with a likelihood ratio of 1.29. CONCLUSIONS: Counting WBC instead of a specific stain for the detection of leukocytes has only a poor sensitivity/specificity for the detection of bacteria. PMID- 16314033 TI - Holmium laser enucleation of the prostate (HoLEP): the endourologic alternative to open prostatectomy. AB - OBJECTIVE: To evaluate HoLEP for patients with enlarged prostate (traditionally treated by open prostatectomy) with long-term follow-up. METHODS: A retrospective analysis of 225 consecutive patients presenting with lower urinary symptoms secondary to benign prostatic hyperplasia with large prostate (>80 cc) who underwent HoLEP. Enucleation time, morcellation time, enucleated tissue weight, catheterization time, hospital stay, voiding outcome parameters, and complications were recorded. RESULTS: Mean preoperative prostate volume was 126+/ 45.1 cc (range 80-351, median 111.2 cc), and resected tissue weight was 86.5 g. Mean follow-up was 31+/-12 months (median 24 months). Mean catheter time and hospital stay were 1.3 and 1.2 days, respectively. Patient symptom scores and peak flow rates were significantly improved immediately after surgery and continued to improve during subsequent follow up. Two patients required intraoperative blood transfusion, and a third patient needed blood transfusion in the early postoperative period for persistent hematuria. Bladder neck contracture and urethral stricture developed in 0.4% and 1.3%, respectively. CONCLUSIONS: HoLEP represents a safe and effective treatment for patients with symptomatic large prostates. It offers patients who traditionally required open prostatectomy the alternative of being treated endoscopically with minimal blood loss, short catheterization time and hospital stay. PMID- 16314034 TI - Laparoscopic prostatectomy for benign prostatic hyperplasia--a six-year experience. AB - PURPOSE: The Authors present their results using laparoscopic prostatectomy in the treatment of large benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: Between March 1999 and March 2005, 60 patients were submitted to laparoscopic prostatectomy with vascular control for large BPH. The demographic, operative period and outcome data were recorded. RESULTS: The average prostate weight was 144.50+/-41.74 gm. Mean operative time was 138.48+/-23.38 minutes and estimated blood loss of 330.98+/-149.52 ml. No patient required transfusions or conversion to open surgery. Post operative complications included one case of septicemia and three cases of prolonged ileum. The most frequent long-term complication was retrograde ejaculation, presented in all patients after 6 months of follow-up. The erectile function was preserved in all those patients who were potent before surgery. No urinary incontinence was reported by patients. CONCLUSIONS: The results demonstrate that resection of large prostatic adenomas can be performed with a laparoscopic approach. The patients had a shorter hospital stay and early return to normal activity. PMID- 16314035 TI - Extended 12-core prostate biopsy increases both the detection of prostate cancer and the accuracy of Gleason score. AB - OBJECTIVE: To evaluate the effect of extended 12-core prostate biopsy in improving the detection rate of prostate cancer and increasing the accuracy of Gleason score. METHODS: This study included 113 patients who underwent TRUS guided lateral sextant biopsy (group I) and 176 patients who underwent extended 12-core biopsy (group II). Inclusion criteria for prostate biopsy were elevated serum PSA levels (>3.0 ng/ml) and/or suspicious digital rectal examination (DRE). RESULTS: Clinical characteristics were similar in both groups. Cancer was detected in 28 (24.8%) and 64 (36.4%) patients in group I and II respectively, chi2=4.26, p=0.039. Among patients with cancer in group I, 14 were treated by radical prostatectomy (RP). The median Gleason sum was 6 (range 3-8) and 7 (range 5-9) for needle and prostatectomy specimens respectively. There was an agreement between the biopsy and prostatectomy Gleason sum in 7 (50%) patients while the biopsy Gleason sum was lower in 7 (50%) cases. Among patients with cancer in group II, 27 were treated by RP. The median and the range of Gleason sum was the same for needle and prostatectomy specimen (median 6, range 4-9). There was an agreement between the biopsy and prostatectomy specimen in 23 (85.2%) patients while the biopsy sum was lower than prostatectomy in 4 (14.8%) patients. The agreement between the biopsy and prostatectomy specimen was significantly higher in group II (82.5%) than group I (50%), Fisher's Exact Test, p=0.026. CONCLUSION: Extended 12-core prostate biopsy significantly increases both the detection rate of prostate cancer and the accuracy of biopsy Gleason score. PMID- 16314036 TI - Metastatic spermatocytic seminoma--an extremely rare disease. AB - Metastatic spermatocytic seminoma is an extremely rare disease with only one documented case in literature. We present another patient with metastatic disease confirmed by histological work-up after laparoscopic retroperitoneal lymph node dissection (L-RPLND). PMID- 16314037 TI - Reinnervation for neurogenic bladder: historic review and introduction of a somatic-autonomic reflex pathway procedure for patients with spinal cord injury or spina bifida. AB - Neurogenic bladder caused by SCI or spina bifida is a major problem. Research in restoring functional micturition has mainly focused on electrical stimulation for many decades with good progress, but it is still not the definitive solution for majority of the SCI patients. An alternative approach has been to investigate restoring innervation to the lower urinary tract after spinal SCI. Different animal and clinical studies were reviewed historically in this article, focused on mainly cross over nerve surgery for reinnervation of the bladder. An artificial somatic-autonomic reflex pathway procedure and its mechanisms were introduced. Clinical application and the satisfactory results of the new procedure were reviewed in details in restoring voluntary bladder control in patients with SCI or spina bifida. PMID- 16314038 TI - Failure to achieve castration levels in patients using leuprolide acetate in locally advanced prostate cancer. AB - OBJECTIVE: In a cross-sectional, retrospective, non-randomised study to investigate the possibility that some patients treated with luteinizing hormone releasing hormone analogues (LHRH analogues) fail to reach castration levels of serum testosterone. METHODS: 40 patients treated with a 3-monthly formulation of leuprolide acetate and continuous use of an oral antiandrogen ("Leu group") and 25 patients treated with a 3-monthly formulation of goserelin acetate and an oral antiandrogen for one month ("Gos group") were identified from our hospital's registry. Serum testosterone was measured during treatment with the respective LHRH-analogue and compared between the two groups. In the Leu group, serum testosterone was measured during week 11 or 12 of treatment. In the Gos group, serum testosterone was assessed during week 23 or 24. RESULTS: Four patients (10%) treated with leuprolide acetate failed to reach the castration level of serum testosterone after treatment with one injection of a three-monthly formulation of leuprolide acetate. All patients treated with goserelin acetate achieved the castration level. CONCLUSION: Although the overwhelming majority of prostate cancer patients during treatment of LHRH analogue achieve serum testosterone values within the castration range, individual patients may fail to reach this therapeutic goal, probably more often during treatment with leuprolide acetate than with goserelin acetate. PMID- 16314039 TI - Medical consumption and costs during a one-year follow-up of patients with LUTS suggestive of BPH in six european countries: report of the TRIUMPH study. AB - OBJECTIVE: To determine the medical consumption and associated treatment costs of patients with LUTS suggestive of BPH. METHODS: A prospective, cross-sectional, observational survey in six European countries: France, Germany, Italy, Poland, Spain and the United Kingdom, with a one-year follow-up of incident and prevalent patients. RESULTS: Treatment costs were estimated for 5,057 patients with a mean age of 66 years and a mean IPSS score at inclusion of 11.5. In 30% of patients watchful waiting was the therapy of choice for the full follow-up period, 57% were prescribed alpha-lockers, 11% finasteride and 10% phytotherapy at any moment during the follow-up (including switches and combination of treatment). Surgery rate was 4.9%. Mean one-year treatment costs were 858 per patient, three quarters of which concerned medication costs. Multivariate regression analysis showed that medication choice, complications and undergoing surgery were associated with higher costs. CONCLUSIONS: Treatment costs for patients with LUTS suggestive of BPH were moderate and largely consisted of medication costs. Daily practice and associated costs varied considerably across the six countries. PMID- 16314040 TI - Relationship of safety culture and process safety. AB - Throughout history, humans have gathered in groups for social, religious, and industrial purposes. As the conglomeration of people interact, a set of underlying values, beliefs, and principles begins to develop that serve to guide behavior within the group. These "guidelines" are commonly referred to as the group culture. Modern-day organizations, including corporations, have developed their own unique cultures derived from the diversity of the organizational interests and the background of the employees. Safety culture, a sub-set of organizational culture, has been a major focus in recent years. This is especially true in the chemical industry due to the series of preventable, safety related disasters that occurred in the late seventies and eighties. Some of the most notable disasters, during this time period, occurred at Bhopal, Flixborough, and Seveso. However, current events, like the September 11th terrorist attacks and the disintegration of the Columbia shuttle, have caused an assessment of safety culture in a variety of other organizations. PMID- 16314041 TI - Surface modification of purified fly ash and application in polymer. AB - With the growing general concern about the pollution by fly ash (FA), there has been global interest in its utilization. Purified FA or FA micro-beads are suitable as polymer filling materials because of their density, good dispersity and fluidity of globular particles. However, FA as a filler has not been widely used up to now on account of low whiteness values and low friction of untreated FA surface. In order to improve the FA quality, a surface modification method by using isothermal heating is proposed in this paper. Preparation of composite fly ash (CFA) in the Ca(OH)(2)-H(2)O-CO(2) system is described. Good coating results on FA surfaces can be achieved under suitable operating parameters. The characteristics of CFA are discussed and analyzed based on data from X-ray diffraction, scanning electron microscopy (SEM), infrared spectra, and BET multiple-point nitrogen adsorption method. Feedstocks with less than 45 microm grain size, 2.86 m(2) g(-1) specific surface area, and 36.68 whiteness value revealed an increase in specific surface area ranging from 8.69 to 10.01 m(2) g( 1) and an increase in whiteness values ranging from 63.67 to 73.13 after coating. A SEM study allowed a detailed determination of the morphology of the surface roughness. Filling tests also show that a rough surface of the CFA enhances contact opportunities and improves the interface between polymer and CFA blended with polypropylene (PP). PMID- 16314042 TI - Evaluation of blends tincal waste, volcanic tuff, bentonite and fly ash for use as a cement admixture. AB - The evaluation of blends tincal waste (TW), fly ash (FA), bentonite (BE), volcanic tuff (VT) for use as a cement admixture was investigated. The properties examined include setting time, expansion, water requirement, specific surface and compressive strength of cement mixtures. The results revealed that the early compressive strength decrease with increasing tincal waste, due to tincal waste increasing initial setting time of the cement. The tincal waste and volcanic tuff of cement mixtures increased and there was reduction in compressive strength. The more the tincal waste increased the greater retardation there was initial setting time this may be attributed to containing high amount B2O3 and MgO content. The tincal waste and fly ash increased with expansion increased. Water requirement increased as the Blaine fineness of the cement mixtures increased. The results obtained were compared with standards and five batches were advised as suitable for the standard. PMID- 16314043 TI - Physico-chemical treatments for removal of recalcitrant contaminants from landfill leachate. AB - In this paper, the technical applicability and treatment performance of physico chemical techniques (individual and/or combined) for landfill leachate are reviewed. A particular focus is given to coagulation-flocculation, chemical precipitation, ammonium stripping, membrane filtration and adsorption. The advantages and limitations of various techniques are evaluated. Their operating conditions such as pH, dose required, characteristics of leachate in terms of chemical oxygen demand (COD) and NH3-N concentration and treatment efficiency are compared. It is evident from the survey of 118 papers (1983-2005) that none of the individual physico-chemical techniques is universally applicable or highly effective for the removal of recalcitrant compounds from stabilized leachate. Among the treatments reviewed in this article, adsorption, membrane filtration and chemical precipitation are the most frequently applied and studied worldwide. Both activated carbon adsorption and nanofiltration are effective for over 95% COD removal with COD concentrations ranging from 5690 to 17,000 mg/L. About 98% removal of NH3-N with an initial concentration ranging from 3260 to 5618 mg/L has been achieved using struvite precipitation. A combination of physico-chemical and biological treatments has demonstrated its effectiveness for the treatment of stabilized leachate. Almost complete removal of COD and NH3-N has been accomplished by a combination of reverse osmosis (RO) and an upflow anaerobic sludge blanket (UASB) with an initial COD concentration of 35,000 mg/L and NH3-N concentration of 1600 mg/L and/or RO and activated sludge with an initial COD concentration of 6440 mg/L and NH3-N concentration of 1153 mg/L. It is important to note that the selection of the most suitable treatment method for landfill leachate depends on the characteristics of landfill leachate, technical applicability and constraints, effluent discharge alternatives, cost effectiveness, regulatory requirements and environmental impact. PMID- 16314044 TI - New aspects of initiation reactivities of energetic materials demonstrated on nitramines. AB - A brief survey is presented of the author's results obtained from studies of the chemical micro-mechanisms of nitramines initiation from the point of view of organic chemistry. The relationships have been presented and discussed between the characteristics of impact and electric spark sensitivities, detonation and thermal decomposition, on the one hand, and (15)N NMR chemical shifts of nitrogen atoms of nitramino groups, on the other. In the case of the impact sensitivity, the said relationships involve the (15)N shifts of the amino nitrogen atoms carrying the nitro group primarily split off from the molecule. In the case of the initiation by shock, heat and electric spark, the (15)N shifts of nitrogen atoms in the primarily split off nitro groups themselves are involved. Also, the relationships are presented between the characteristics of thermal reactivity and values of the electronic charges at the nitro groups that are primarily split off. It has been stated that the chemical micro-mechanisms of primary fission processes of molecules of nitramines in the initiation by mechanical stimuli (inclusive the detonation course) and electric spark should be the same as in the case of their low-temperature thermal decomposition. It has been found that the electron structure and close neighbourhood of nitrogen atom of the primarily leaving nitro group is a dominant factor in initiation by shock, electric spark and heat. In the case of initiation by impact a key role plays characteristics of amino nitrogen atoms which are carriers of these most reactive nitro groups. Also mentioned is relevance of the modified Evans-Polanyi-Semenov relationship. On the basis of the findings presented it also has been stated that the detonation transformation itself of the nitramines should be preceded by an induction period. PMID- 16314045 TI - Utilizing the Human, Machine, and Environment Matrix in investigations. AB - "How did we get into this situation?" How many times has this question been asked at the outset of an investigation, or more importantly, at the completion of an investigation? If the answer is not readily and thoroughly apparent, the investigation is not complete. Subsequently, those who will have the responsibility for correction of the conditions leading to the incident will not have all the information necessary to properly complete their task. For many years, in many writings, the Human/Machine interaction and its impact on process design has been discussed. The same impact should be examined when performing incident investigations. Consideration of the interaction of human and machine along with the environment in which they are used has long been recommended by the National Safety Council, in both design and investigation. PMID- 16314046 TI - Creatine and neurotrophin-4/5 promote survival of nitric oxide synthase expressing interneurons in striatal cultures. AB - Nitric oxide (NO) mediates a variety of physiological functions in the central nervous system and acts as an important developmental regulator. Striatal interneurons expressing neuronal nitric oxide synthase (nNOS) have been described to be relatively spared from the progressive cell loss in Huntington's disease (HD). We have recently shown that creatine, which supports the phosphagen energy system, induces the differentiation of GABAergic cells in cultured striatal tissue. Moreover, neurotrophin-4/5 (NT-4/5) has been found to promote the survival and differentiation of cultured striatal neurons. In the present study, we assessed the effects of creatine and NT-4/5 on nNOS-immunoreactive (-ir) neurons of E14 rat ganglionic eminences grown for 1 week in culture. Chronic administration of creatine [5mM], NT-4/5 [10ng/ml], or a combination of both factors significantly increased numbers of nNOS-ir neurons. NT-4/5 exposure also robustly increased levels of nNOS protein. Interestingly, only NT-4/5 and combined treatment significantly increased general viability but no effects were seen for creatine supplementation alone. In addition, NT-4/5 and combined treatment resulted in a significant larger soma size and number of primary neurites of nNOS-ir neurons while creatine administration alone exerted no effects. Double-immunolabeling studies revealed that all nNOS-ir cells co localized with GABA. In summary, our findings suggest that creatine and NT-4/5 affect differentiation and/or survival of striatal nNOS-ir GABAergic interneurons. These findings provide novel insights into the biology of developing striatal neurons and highlight the potential of both creatine and NT 4/5 as therapeutics for HD. PMID- 16314047 TI - Growth inhibitory factor (GIF) can protect from brain damage due to stab wounds in rat brain. AB - We examined the effect of growth inhibitory factor (GIF), also called metallothionein-III (MT-III), in brain damage using a stab wound model. The administration of 3 microM purified rat GIF (prGIF) provided significantly improved brain repair compared with controls, whereas the administration of 15 microM prGIF reduced brain repair compared with controls. To maintain the continuous effect of GIF, we generated an adenoviral vector encoding rat GIF and the myc epitope (AxCArGIFM) and administered an appropriate amount (1 x 10(8) pfu) of AxCArGIFM on the basis of the optimal dosage determined in a previous study on avulsion of the facial nerve. The administration of AxCArGIFM provided significantly improved histological and biochemical parameters of brain repair compared with controls administered AxCALacZ (adenovirus encoding bacterial beta galactosidase gene as a reporter; 1 x 10(8) pfu). These results show that GIF can protect from brain damage in certain appropriate conditions in vivo and in vitro. The optimal dosage is very important for the treatment in vivo, particularly that for GIF. Our findings show the double-edged effects of GIF. MTs including MT-III are promising as therapeutic agents not only for tissue repair following acute brain injury, but also for some neurodegenerative diseases because they have multifunctional potential including anti-oxidation effects and may have some effect on neurogenesis. PMID- 16314048 TI - Event related fMRI studies of voluntary and inhibited eye blinking using a time marker of EOG. AB - Electrooculogram (EOG) measurements, along with infrared measurements, are commonly used to record eye blinking during functional magnetic resonance imaging (fMRI). We report herein, on the use of EOG in measuring voluntary and inhibited eye blinking during echo planar imaging (EPI) in an MR scanner. The inhibited eye blinking occurred during the period, in which subjects were requested not to blink their eyes. After the removal of gradient-field induced artifacts from the EOG signal, the waveform of the EOG clearly showed both voluntary and inhibited eye blinking. Using these data, each voluntary or inhibited eye-blinking event was used as the temporal cue for an event related fMRI. Activation of the bilateral parahippocampal, precentral gyrus and left supplementary motor area was observed for voluntary eye blinking, whereas the medial/superior frontal, precentral, cingulate, precuneus, and superior temporal gyrus appears to be involved in inhibited eye blinking. Based on these experimental results, we propose that the precentral gyrus is responsible for both voluntary and inhibited eye blinking. The parietal area (precuneus and superior temporal gyrus) appears to be exclusively related to inhibited eye blinking. PMID- 16314049 TI - Correlates of motivation to quit smoking among alcohol dependent patients in residential treatment. AB - Substance use and smoking co-occur at high rates and substance abusers smoke more and have greater difficulty quitting smoking compared to the general population. Methods of increasing smoking cessation among alcoholics are needed to improve their health. This study investigated predictors of motivation to quit smoking among patients early in residential treatment for substance abuse. The 198 alcohol dependent patients were participating is a larger smoking study at an inner-city residential substance abuse treatment program. Motivation was measured by the Contemplation Ladder. A hierarchical multiple regression was conducted to assess whether perceived barriers to smoking cessation and self-efficacy about quitting were associated with motivation to quit smoking independent of the influence of degree of tobacco involvement, substance use, and comorbid depressive symptoms. Motivation was higher with longer previous smoking abstinence, fewer barriers to quitting, and greater self-efficacy but was not influenced by smoking rate, dependence, or gender. While the combination of alcohol and drug use, alcohol and drug problem severity, and depressive symptoms predicted motivation, no one of these variables was significant. Since barriers to change and self-efficacy are potentially modifiable in treatment, these could be salient targets for intervention efforts. This could be integrated into treatment by assessing barriers and providing corrective information about consequences and methods of overcoming barriers and by providing coping skills to increase confidence in one's ability to quit smoking. PMID- 16314050 TI - The bone-related Zn finger transcription factor Osterix promotes proliferation of mesenchymal cells. AB - Osterix is a bone-related transcription factor that functions genetically downstream of Runx2, which controls both growth and differentiation in osteoblasts. Here we assessed the biological function of Osterix in mesenchymal cells that are not normally committed to the osteogenic lineage. Stably transfected NIH3T3 fibroblasts that express exogenous Osterix were examined for their ability to convert into osteoblastic cells by analyzing gene expression profiles of bone phenotype related markers, as well as by measuring bone nodule formation and cell proliferation. Forced expression of Osterix stimulates osteopontin gene expression but not the expression or activity of other bone related markers, including collagen type I, alkaline phosphatase, osteocalcin, or osteonectin. Moreover, cells stably expressing Osterix do not induce bone nodule formation. Strikingly, both polyclonal and monoclonal cells expressing Osterix exhibit enhanced proliferation. Collectively, these results indicate that Osterix is insufficient to establish osteogenic lineage commitment, perhaps due to the ability of Osterix to promote cell growth. We propose that regulatory pathways operating upstream of or in parallel with Osterix are required for osteogenic conversion of uncommitted mesenchymal cells. PMID- 16314051 TI - Xenopus apyrase (xapy), a secreted nucleotidase that is expressed during early development. AB - We have characterized a cDNA encoding a Xenopus laevis apyrase (XAPY) that is expressed during embryogenesis. XAPY is highly homologous to two recently described mammalian apyrases, human SCAN-1 and rat Ca2+-NDPase, and to a lesser extent the salivary apyrase of the blood-feeding arthropod Cimex lectularis. RT PCR analysis shows that Xapy is expressed at all the developmental stages tested, from oocytes through to tadpoles. Xapy transcripts are widely distributed in the embryo, but from late neurulae through to late tailbud stages they are highly enriched in the cement gland, an adhesive organ in the epidermis of the head. When expressed in HEK 293 cells, XAPY is largely retained in the endoplasmic reticulum, although some is also secreted. XAPY conditioned media hydrolyses UDP and UTP, confirming that it is a functional apyrase. PMID- 16314052 TI - EVI1 and hematopoietic disorders: history and perspectives. AB - The ecotropic viral integration site 1 (EVI1) gene was identified almost 20 years ago as the integration site of an ecotropic retrovirus leading to murine myeloid leukemia. Since its identification, EVI1 has slowly been recognized as one of the most aggressive oncogenes associated with human leukemia. Despite the effort of many investigators, still very little is known about this gene. The mechanism by which EVI1 operates in the transformation of hematopoietic cells is not known, but it is clear that EVI1 upregulates cell proliferation, impairs cell differentiation, and induces cell transformation. In this review, we summarize the biochemical properties of EVI1 and the effects of EVI1 in biological models. PMID- 16314053 TI - Critical molecular switches involved in BMP-2-induced osteogenic differentiation of mesenchymal cells. AB - Bone morphogenetic protein (BMP)-2 strongly induces bone formation. Introduction of the protein in muscle tissue results in ectopic bone formation. Similarly, BMP 2 treatment also stimulates the in vitro transdifferentiation of myogenic cells to osteogenic cells. The establishment of an in vitro model system has enabled the investigation of intracellular events including BMP receptor activation, BMP 2-induced R-Smad activation, and kinase activation, and the role of osteogenic transcription factors, such as Runx2, Osx, Dlx5, and Msx2. Many reviews have addressed events downstream of BMP-receptor binding but few deal with molecular cascades involved in BMP-2-induced osteogenesis. We focus on critical molecular switches, especially transcription factors, and several kinase pathways involved in osteogenic differentiation. PMID- 16314054 TI - Sequence context analysis of 8.2 million single nucleotide polymorphisms in the human genome. AB - We analyzed n-mers (n=3-8) in the local environment of 8,249,446 human SNPs and compared their distribution with that in the genome reference sequences. The results revealed that the short sequences, which contained at least one CpG dinucleotide, occurred more frequently in the local SNP sequences than in the genome sequences. To exclude the hypermutability effect of the methylated CpG dinucleotides on the sequence context of SNPs, we examined the distribution patterns for each of the six categories of substitution. We observed the similar pattern (i.e., CpG-containing n-mers vs. non-CpG-containing n-mers) in SNP categories A/G, C/T and C/G but the opposite pattern in category A/T. We next identified 34,928 putative CpG islands in the human genome and located 133,591 SNPs within these islands. In the CpG islands, CpG SNPs were 3.92-fold less prevalent relative to the presence of CpG dinucleotides. Conversely, in the human genome, the frequency of CpG dinucleotides at the polymorphic sites was 6.09 times that in the genome reference sequences. These results support the previous views of mutational suppression at the CpG sites in the CpG islands and hypermutability of the methylated CpG dinucleotides that are prevalent in the non CpG island sequences in the human genome. Our study represents a comprehensive investigation of the sequence context of SNPs in the human genome and in human CpG islands. PMID- 16314055 TI - Utilization of a toxicogenomic biomarker for evaluation of chemical-induced glutathione deficiency in rat livers across the GeneChip data of different generations. AB - Previously, we reported 69 probe sets (GSH probe sets) of RG U34A GeneChip that were useful for the evaluation of chemical-induced glutathione depletion in rat livers. The aim of the present study was to investigate whether these probe sets could be applied to the analysis of RAE 230A GeneChip data. Since a straightforward data comparison of RG U34A and RAE 230A GeneChips could not overcome the generation-dependent discrepancy in signal profiles, we tried two methods to improve the data compatibility between the two GeneChips. First, we re calculated the signal values by excluding the probes with poor-overlapping sequences between the two GeneChips, but the data compatibility did not improve from the view point of Spearman's and Pearson's correlation coefficients. On the other hand, the PCA result demonstrated that an adjustment of the baseline signal level between the RG U34A and RAE 230A GeneChip data on vehicle-treated rats dramatically improved the data compatibility, suggesting that the GSH probe sets identified from RG U34A GeneChip data can be utilized in RAE 230A GeneChip data as well. Such a baseline adjustment of signal data is an easy and practical way to utilize biomarkers across GeneChip data of different generations. PMID- 16314056 TI - Mitoxantrone-loaded BSA nanospheres and chitosan nanospheres for local injection against breast cancer and its lymph node metastases. II: Tissue distribution and pharmacodynamics. AB - Bovine serum albumin (BSA) and chitosan (CS) nanospheres of mitoxantrone (MTO) were comparatively evaluated in terms of tissue distribution, acute toxicity and therapeutic efficiency against breast cancer and its lymph node metastases. After local injection in rats, MTO nanospheres showed a slower elimination rate and a much higher drug concentration in lymph nodes compared with MTO solution, and a lower drug concentration in other tissues. There was no observed acute toxicity to the main tissues of Kunming mice after local injection of MTO-BSA-NS. Mild toxicity to liver and lung was observed for MTO-CS-NS, but, for MTO solution, severe toxicity to liver and lung and much lower number of white blood cells were observed. Human MCF-7 breast cancer in nude mice and animal model of P388 lymph node metastases in Kunming mice were applied to investigate the therapeutic efficiency. The inhibition rate of the nanospheres against breast cancer was much higher than that of MTO solution, and lymph node metastases were efficiently inhibited by the nanospheres, especially MTO-BSA-NS. PMID- 16314057 TI - Anti-inflammatory activity of Croton cuneatus aqueous extract. AB - The aqueous extract of Croton cuneatus Klotz. (Euphorbiaceae), was tested for its antinociceptive effects using chemical and thermal test models in mice. Anti inflammatory activity was determined in Sprague-Dawley rats in a model of acute plantar inflammation induced by bovine serum albumin. Croton cuneatus aqueous extract at doses of 7 mg/kg showed a significant anti-inflammatory effect compared with commonly used non-stereoidal drugs as ketoprofen, sodium diclofenac and ASA (acetylsalicylic acid). PMID- 16314058 TI - In vitro protective effects of salvianolic acid B on primary hepatocytes and hepatic stellate cells. AB - The production of reactive oxygen species (ROS) is believed to be involved in liver injury and hepatic fibrosis. Activation of hepatic stellate cells (HSCs) is a key feature of liver fibrosis. Salvia miltiorrhiza is a traditional Chinese herb used in the treatment of cardiovascular and liver diseases to resolve stasis. The effects of salvianolic acid B (Sal B), a major component of Salvia miltiorrhiza, on oxidative damage include free radical DPPH scavenging, malondialdehyde (MDA) formation and ROS generation in primary rat hepatocytes and HSCs, and on alpha-SMA, and collagen expression in transforming growth factor beta1 (TGF-beta1)-stimulated HSCs were examined. Results indicated that Sal B scavenged DPPH potently with an IC50 2.2+/-0.2 microg/ml (3.06+/-0.3 microM), inhibited lipid peroxidation and eliminated ROS accumulation in a concentration dependent manner on primary rat hepatocytes and HSCs. Sal B also reduced alpha SMA and collagen synthesis and deposition in HSCs, and had no direct cytotoxicity on both hepatocytes and HSCs. Our results suggest that Sal B ameliorated oxidative damage and eliminated ROS accumulation in hepatocytes, and attenuated HSC activation, potentially conferring hepatoprotective and anti-fibrogenic effects. PMID- 16314059 TI - Inhibition of gastric H+,K+-ATPase activity by flavonoids, coumarins and xanthones isolated from Mexican medicinal plants. AB - Medicinal plants are commonly used in Latin American folk medicine for the treatment of gastric problems. In order to understand the properties of some of their chemical constituents, four natural xanthones, an acetylated derivative, two coumarins (mammea A/BA and mammea C/OA) isolated from Calophyllum brasiliense Cambess and two flavonoids (minimiflorin and mundulin) isolated from Lonchocarpus oaxacensis Pittier, and the chalcone lonchocarpin isolated from Lonchocarpus guatemalensis Benth were tested for their activities on gastric H+,K+-ATPase isolated from dog stomach. All the compounds tested inhibited H+,K+-ATPase activity with varied potency. The xanthones inhibited the H+,K+-ATPase with IC50 values ranging from 47 microM to 1.6 mM. Coumarins inhibited H+,K+-ATPase with IC50 values of 110 and 638 microM. IC50 values for the flavonoids ranged from 9.6 to 510 microM among which minimiflorin was the most potent. The results suggest that H+,K+-ATPase is sensitive to inhibition by several types of structurally different natural compounds. The potency of the effects on gastric H+,K+-ATPase depends on the presence, position and number of hydroxyls groups in the molecule. Collectively, these results suggest a potential for important pharmacological and toxicological interactions by these types of natural products at the level of H+,K+-ATPase which may explain, at least in part, the gastroprotective properties, indicated by traditional medicine, of the plants from which these compounds were isolated. PMID- 16314060 TI - Population genetic study in two Transylvanian populations using forensically informative autosomal and Y-chromosomal STR markers. AB - Our study provides population genetic data on two population samples collected in a Hungarian speaking region of Transylvania, Romania. Allele frequency and profile databases were generated on 17 autosomal STR loci (D2S1338, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D19S433, D21S11, VWA, FGA, TH01, TPOX, CSF1PO, Penta E and Penta D) as well as at the 12 European Y-STR extended haplotype loci (DYS19, DYS389-I/II, DYS390, DYS391, DYS392, DYS393, DYS385 loci, DYS437, DYS438 and DYS439). Data were compared to a Central Hungarian (Budapest region) population sample [B. Egyed, S. Furedi, M. Angyal, L. Boutrand, A. Vandenberghe, J. Woller, Z. Padar, Analysis of eight STR loci in two Hungarian populations, Forensic Sci. Int. 113 (2000) 25-27] that was used as a reference group of the Hungarian population. Calculating the F(ST) indices and with the pairwise comparisons of interpopulation molecular variance (AMOVA) the two populations from Transylvania could be fit into the Hungarian population data showing less substructuring effects as compared to the previous findings in Hungary [B. Egyed, S. Furedi, M. Angyal, L. Boutrand, A. Vandenberghe, J. Woller, Z. Padar, Analysis of eight STR loci in two Hungarian populations, Forensic Sci. Int. 113 (2000) 25-27; B. Egyed, S. Furedi, M. Angyal, I. Balogh, L. Kalmar, Z. Padar, Analysis of the population heterogeneity in Hungary using fifteen forensically informative STR markers, Forensic Sci. Int. 158 (2005) 244-249]. PMID- 16314061 TI - Application of atomic absorption spectroscopy for detection of multimetal traces in low-voltage electrical marks. AB - Application of atomic absorption spectroscopy to detect multimetal traces in injured skin is a promising tool for investigation of fatalities caused by electrocution. The present paper is aimed at testing the reliability of this method for metal traces detection in electric current marks and is focused on study of peculiarities of metal penetration into the skin exposed to a current impact. Bare aluminum wire, tin-lead coated copper multistrand wire, and zinc plated steel rope were used to make electrical marks on pig skin. It is demonstrated that amount of copper, zinc, lead, and iron may serve as statistically reliable indicators for the type of wire, which caused the electrical mark, in spite of the background content of these metals in the skin without injury. Different penetration rates for different metals contained in the wire inflicting an electrical mark were observed. PMID- 16314062 TI - Non-contrast computerized tomography (NCCT) and dynamic renal scintigraphy (DRS) in the patients with refractory renal colic. AB - OBJECTIVES: To assess the importance of combined use of non-contrast computerized tomography (NCCT) and dynamic renal scintigraphy (DRS) in evaluation of patients with refractory flank pain in the emergency department. METHODS: The study involved 64 consecutive patients with refractory renal colic. All patients were evaluated with plain abdominal films kidneys, ureters and bladder (KUB), NCCT and DRS. We assessed the accuracy of different diagnostic procedures and their combinations; in addition, we determined their importance for different steps of evaluation. RESULTS: Urololithiasis was diagnosed in 76.6% (n=49) of the patients. Twenty-nine percent of calculi were > 4 mm. Surgical intervention were performed in 20 patients (40.8%). A combination of NCCT plus DRS yielded the greatest sensitivity (96%) in establishing final diagnosis, however clinical, laboratory and KUB data in combination with DRS, yielded greater specificity (93%) and PPV (97%). Sex (male), WBC (mean 10.2 x 10(3) +/- 3.1) and KUB (calculus > 4 mm) were chosen in the three-step multi-variant analysis, while only male sex was found to be the strongest predictor (p<0.056) of necessity to perform NCCT. In making decision for definitive treatment NCCT and DRS provided the most important information about stone size and obstruction (kappa=0.734, p<0.001 and kappa=0.625, p<0.001), while DRS was selected as the most important diagnostic procedure in the emergency department (kappa=0.527, p<0.001). CONCLUSIONS: In the emergency department, DRS combined with the results of clinical investigation may indicate candidates for hospitalization with emergency intervention. Immediate NCCT must be strongly considered in men with WBC> or =10 x 10(3) and calculi > 4 mm on the KUB. PMID- 16314063 TI - Whole brain magnetization transfer histogram analysis of pediatric acute lymphoblastic leukemia patients receiving intrathecal methotrexate therapy. AB - BACKGROUND AND PURPOSE: The purpose of this prospective study was to evaluate the hypothesis that magnetization transfer ratio (MTR) histogram analysis of the whole brain could detect early and subtle brain changes nonapparent on conventional magnetic resonance imaging (MRI) in children with acute lymphoblastic leukemia (ALL) receiving methotrexate (MTX) therapy. MATERIALS AND METHODS: Subjects in this prospective study comprised 10 children with ALL (mean age, 6 years; range, 0-16 years). In addition to conventional MRI, magnetization transfer images were obtained before and after intrathecal and intravenous MTX therapy. MTR values were calculated and plotted as a histogram, and peak height and location were calculated. Differences in peak height and location between pre and post-MTX therapy scans were statistically analyzed. Conventional MRI was evaluated for abnormal signal area in white matter. RESULTS: MTR peak height was significantly lower on post-MTX therapy scans than on pre-MTX therapy scans (p = 0.002). No significant differences in peak location were identified between pre- and post-chemotherapy imaging. No abnormal signals were noted in white matter on either pre- or post-MTX therapy conventional MRI. CONCLUSIONS: This study demonstrates that MTR histogram analysis allows better detection of early and subtle brain changes in ALL patients who receive MTX therapy than conventional MRI. PMID- 16314064 TI - Development of an extraction method for mycobacterial metabolome analysis. AB - As a prerequisite for studying the intracellular metabolome of mycobacteria, several methods were evaluated for efficient breakage of the cell using Mycobacterium bovis (BCG) as a model microorganism. Several pulping methods, treating with an Ultra-Turax, deep-freezing in liquid nitrogen followed by mechanical grinding, sonicating with probe head or cup horn and bead beating prior to solvent extraction were applied and compared. Gravimetry, electron microscopy, and nuclear magnetic resonance spectrometry were used to analyze the extracts. All analytical methods prove that sonicating is superior to mechanical grinding of deep-frozen cells. Two methods indicated that sonicating with a probe head enhances the efficiency of cell disruption compared to sonicating with a cup horn. The highest extract yield and chemical diversity were achieved by a combination of mechanical grinding and sonicating. Within the scope of a metabolomic analysis, the method of choice to treat mycobacterial cells is a combination of deep-freezing in liquid nitrogen and mechanical grinding followed by sonicating with a probe head. PMID- 16314065 TI - Modification of the ultrafiltration technique to overcome solubility and non specific binding challenges associated with the measurement of plasma protein binding of corticosteroids. AB - Plasma protein binding (PPB) methodology suitable for application in the lead optimisation of a corticosteroid series known to demonstrate non-specific binding (NSB) and poor solubility has been established. The method involved a modification to standard ultrafiltration (UF) techniques. In parallel with each experimental plasma sample, a control plasma sample was also processed by ultrafiltration. The retentate from experimental and control plasma samples were mixed back into the filtrate of the partner sample. The resulting regenerated plasma samples, one representing the experimental filtrate and one representing the experimental retentate, were then analysed by LC/MS/MS. Varying degrees of NSB were demonstrated with a number of corticosteroids, and this effect was eliminated using the modified method. Validation using a panel of established corticosteroids showed good agreement with published PPB figures. The published PPB figure for fluticasone propionate (FP) was, however, found to be an underestimate, and this was subsequently confirmed, at clinically relevant plasma concentrations, to be 99.3%. The modified method was particularly suited to lead optimisation because it provided samples in a consistent matrix compatible with standard high throughput LC/MS/MS analysis. PMID- 16314066 TI - Determination of ruthenium in pharmaceutical compounds by graphite furnace atomic absorption spectroscopy. AB - A graphite furnace atomic absorption (GFAA) spectrometric method for the determination of ruthenium (Rh) in solid and liquid pharmaceutical compounds has been developed. Samples are dissolved or diluted in dimethyl sulfoxide (DMSO) without any other treatment before they were analyzed by GFAA with a carefully designed heating program to avoid pre-atomization signal loss and to achieve suitable sensitivity. Various inorganic and organic solvents were tested and compared and DMSO was found to be the most suitable. In addition, ruthenium was found to be stable in DMSO for at least 5 days. Spike recoveries ranged from 81 to 100% and the limit of quantitation (LOQ) was determined to be 0.5 microg g(-1) for solid samples or 0.005 microg ml(-1) for liquid samples based a 100-fold dilution. The same set of samples was also analyzed by ICP-MS with a different sample preparation method, and excellent agreement was achieved. PMID- 16314067 TI - MIP-1beta, a novel biomarker for in vitro sensitization test using human monocytic cell line. AB - In order to seek a novel biomarker for predicting skin sensitization, changes in the gene expression profile of THP-1 cells on exposure to 2,4 dinitrochlorobenzene (DNCB), p-phenylenediamine (pPD) and nickel sulfate (Ni) were assessed using oligo-DNA microarrays. While the change in gene expression varied depending on the sensitizers, up-regulation of MIP-1 beta mRNA expression was detected in both DNCB-treated and Ni-treated THP-1 cells. This finding was validated by RT-PCR and confirmed at the protein level by ELISA. Secretion of MIP 1 beta from THP-1 was detected after 24-h treatment with sensitizers such as DNCB, Ni, 2-mercaptobenzothiazole (2-MBT) and cobalt sulfate (Co), while pPD and non-sensitizers such as sodium dodecyl sulfate (SDS) and benzalkonium chloride (BC) had no effect. The use of both MIP-1 beta production and CD86 expression as criteria reduced the number of false-negatives, and the results were in good agreement with those of in vivo assays. MIP-1 beta may be useful as a novel biomarker in in vitro sensitization assay using THP-1 cells, either alone or in combination with known markers. PMID- 16314068 TI - Volatile anaesthetic halothane causes DNA damage in A549 lung cells. AB - The present study was performed to elucidate the extent of damage and the ability of lung epithelial cells to recover or to undergo apoptosis after in vitro treatment with the volatile anaesthetic halothane. The results obtained from the comet assay clearly show that halothane, applied at 3.0mM concentration, causes DNA and cell damage. Cells exhibited nuclear fragmentation and budding early after treatment and these events gradually increased during the next few days. The presence of a large number of mini-comets after single cell gel electrophoresis was found to represent apoptotic bodies with fragmented DNA. Our results demonstrate apoptosis-like changes after in vitro exposure of A549 cells to the volatile anaesthetic halothane. The majority of the affected cells did not recover and were directed to cell death. PMID- 16314069 TI - Anticlastogenic effects of black tea polyphenols theaflavins and thearubigins in human lymphocytes in vitro. AB - Black tea accounts for nearly 80% of total World tea production. It contains dimeric flavanols and polymeric polyphenols known as theaflavins (TF) and thearubigins (TR). TR is exclusively present in black tea. On the basis of our previous potent antimutagenic and anticlastogenic effects of TF and TR in vitro in bacterial system and in vivo in mouse bone marrow cells, we have decided to extend our study in human cells in vitro. This study investigated the anticlastogenic effects of black tea polyphenols TF and TR as measured by chromosomal aberrations (CA) and micronuclei formation (MN) against two known mutagens/carcinogens i.e. benzo[a]pyerne (B[a]P) and aflatoxin B1(AFB1) with S9 activation. A significant decrease in both CA and MN were observed in the human lymphocyte cultures treated with either TF or TR pretreated with either B[a]P or AFB1 (250, 500, 1000 microg/ml) when compared with B[a]P or AFB1 treated cultures alone. TF shows more protective effects than TR in this in vitro system. These results indicate that both TF and TR have significant anticlastogenic effects in vitro in human lymphocytes. PMID- 16314070 TI - In vivo relevance of two critical levels for NAD(P)H:quinone oxidoreductase (NQO1)-mediated cellular protection against electrophile toxicity found in vitro. AB - NAD(P)H:quinone oxidoreductase (NQO1)-mediated detoxification of quinones is suggested to be involved in cancer prevention. In the present study, using transfected CHO cells, it was demonstrated that the relation between NQO1 activity and the resulting protection against the cytotoxicity of menadione shows a steep dose-response curve revealing a 'lower protection threshold' of 0.5mumol DCPIP/min/mg protein and an 'upper protection threshold' at 1mumol DCPIP/min/mg protein. In an additional in vivo experiment it was investigated how both in vitro critical activity levels of NQO1, relate to NQO1 activities in mice and man, either without or upon induction of the enzyme by butylated hydroxyanisol (BHA) or indole-3-carbinol (I(3)C). Data from an experiment with CD1 mice revealed that base-line NQO1 levels in liver, kidney, small intestine, colon and lung are generally below the observed 'lower protection threshold' in vitro, this also holds for most human tissue S-9 samples. To achieve NQO1 levels above this 'lower protection threshold' will require 5-20 fold NQO1 induction. Discussion focuses on the relevance of the in vitro NQO1 activity thresholds for the in vivo situation. We conclude that increased protection against menadione toxicity can probably not be achieved by NQO1 induction but should be achieved by other mechanisms. Whether this conclusion also holds for other electrophiles and the in vivo situation awaits further definition of their NQO1 protection thresholds. PMID- 16314071 TI - Single linkage group per chromosome genetic linkage map for the horse, based on two three-generation, full-sibling, crossbred horse reference families. AB - A genetic linkage map of the horse consisting of 742 markers, which comprises a single linkage group for each of the autosomes and the X chromosome, is presented. The map has been generated from two three-generation full-sibling reference families, sired by the same stallion, in which there are 61 individuals in the F2 generation. Each linkage group has been assigned to a chromosome and oriented with reference to markers mapped by fluorescence in situ hybridization. The average interval between markers is 3.7 cM and the linkage groups collectively span 2772 cM. The 742 markers comprise 734 microsatellite and 8 gene based markers. The utility of the microsatellite markers for comparative mapping has been significantly enhanced by comparing their flanking sequences with the human genome sequence; this enabled conserved segments between human and horse to be identified. The new map provides a valuable resource for genetically mapping traits of interest in the horse. PMID- 16314072 TI - Annotating nonspecific SAGE tags with microarray data. AB - SAGE (serial analysis of gene expression) detects transcripts by extracting short tags from the transcripts. Because of the limited length, many SAGE tags are shared by transcripts from different genes. Relying on sequence information in the general gene expression database has limited power to solve this problem due to the highly heterogeneous nature of the deposited sequences. Considering that the complexity of gene expression at a single tissue level should be much simpler than that in the general expression database, we reasoned that by restricting gene expression to tissue level, the accuracy of gene annotation for the nonspecific SAGE tags should be significantly improved. To test the idea, we developed a tissue-specific SAGE annotation database based on microarray data (). This database contains microarray expression information represented as UniGene clusters for 73 normal human tissues and 18 cancer tissues and cell lines. The nonspecific SAGE tag is first matched to the database by the same tissue type used by both SAGE and microarray analysis; then the multiple UniGene clusters assigned to the nonspecific SAGE tag are searched in the database under the matched tissue type. The UniGene cluster presented solely or at higher expression levels in the database is annotated to represent the specific gene for the nonspecific SAGE tags. The accuracy of gene annotation by this database was largely confirmed by experimental data. Our study shows that microarray data provide a useful source for annotating the nonspecific SAGE tags. PMID- 16314073 TI - A knock-in mouse model of congenital erythropoietic porphyria. AB - Congenital erythropoietic porphyria (CEP) is a recessive autosomal disorder characterized by a deficiency in uroporphyrinogen III synthase (UROS), the fourth enzyme of the heme biosynthetic pathway. The severity of the disease, the lack of specific treatment except for allogeneic bone marrow transplantation, and the knowledge of the molecular lesions are strong arguments for gene therapy. An animal model of CEP has been designed to evaluate the feasibility of retroviral gene transfer in hematopoietic stem cells. We have previously demonstrated that the knockout of the Uros gene is lethal in mice (Uros(del) model). This work describes the achievement of a knock-in model, which reproduces a mutation of the UROS gene responsible for a severe UROS deficiency in humans (P248Q missense mutant). Homozygous mice display erythrodontia, moderate photosensitivity, hepatosplenomegaly, and hemolytic anemia. Uroporphyrin (99% type I isomer) accumulates in urine. Total porphyrins are increased in erythrocytes and feces, while Uros enzymatic activity is below 1% of the normal level in the different tissues analyzed. These pathological findings closely mimic the CEP disease in humans and demonstrate that the Uros(mut248) mouse represents a suitable model of the human disease for pathophysiological, pharmaceutical, and therapeutic purposes. PMID- 16314074 TI - Fractionation of chromosome 15 with an affinity-based approach using magnetic beads. AB - The two main shortcomings of the state-of-the-art method of sorting chromosomes, specificity and the efficiency of fractionating a significant amount of chromosomes, are addressed by this work in the design of a massively parallel approach using magnetic beads binding to a chromosome-specific DNA probe. In an attempt to isolate human chromosome 15 from a lymphoblastoid cell line, a chromosome 15 centromere-specific DNA probe with a fluorescent tag attached was reacted with the chromosomes. Magnetic beads bound to anti-FITC antibody were reacted with the labeled pool of chromosomes and separated by exposure to a magnetic field. The specificity of the fractionated pool was verified by performing fluorescence in situ hybridization on the isolated pool. The chromosome of interest could be enriched to about 75% within a maximum of 3-4 days, regardless of the amount of material. PMID- 16314075 TI - Double-blind study on the effects of topical anesthesia on laryngeal secretions. AB - The application of topical anesthesia to the oropharynx is a common clinical practice during oral and nasal laryngoscopy. Clinically, questions have been raised about whether topical anesthesia alters laryngeal secretions, which distorts clinical impressions. A double-blind, placebo controlled design was employed to address this issue. Ten premenopausal women with healthy vocal folds and 10 premenopausal women with phonotraumatic lesions underwent oral videolaryngoscopic examinations on subsequent days under both anesthesia and placebo conditions, in counterbalanced order. Video segments were rated by three judges. Dependent variables were balling and pooling of secretions, as previously described in the literature. Statistical analyses failed to reveal any clear effect of topical anesthesia on either secretion balling or pooling for the collapsed data set, but one cannot exclude changes in individual cases. Moreover, there was no evidence that secretions were differentially affected by anesthesia across subject groups. Null results in this data set replicate and extend previously reported findings by other authors. An incidental but potentially interesting finding was that the order of treatment condition (anesthesia versus placebo first) seemed relevant for secretions: Subjects who received the anesthesia condition first tended to show more secretion balling in general, as compared with subjects who received the placebo condition first. Speculation is entertained regarding possible physiological pathways for these incidental findings, which could be relevant for some clinical practice. PMID- 16314076 TI - Low frequency repetitive transcranial magnetic stimulation improves source monitoring deficit in hallucinating patients with schizophrenia. AB - Auditory hallucinations have been associated with a disruption in monitoring one's own speech suggesting an autonoetic agnosia in schizophrenia. This deficit can be measured by a source monitoring task. Low frequency transcranial magnetic stimulations (rTMS) applied to the left temporoparietal cortex can inhibit cortical areas involved both in autonoetic agnosia (which means 'the inability to identify self-generated mental events') and in auditory hallucinations (AH) phenomena. Although improvements in AH have been repeatedly reported following rTMS treatment, effects on autonoetic agnosia measured by source monitoring have never been investigated. We aimed to investigate the relation between improvements in AH and source monitoring performance after rTMS treatment. Twenty four right-handed refractory schizophrenic patients with hallucinations randomly received sham or active 10.0001-Hz rTMS to the left temporoparietal cortex and performed 2 source monitoring tasks requiring discrimination between silent- and overt-reading words before and after rTMS sessions. Compared to sham, active rTMS significantly improved AH. Source monitoring performances and the improvements tended to correlate, which would support a specific relation between autonoetic agnosia and auditory hallucinations. PMID- 16314077 TI - Validity of the CAGE questionnaire in schizophrenic patients with alcohol abuse and dependence. AB - The objective of the study was to assess the sensitivity and the specificity of the CAGE questionnaire in 114 schizophrenic patients. The Composite International Diagnostic Interview was used to assess the DSM-III-R diagnosis of abuse of or dependence on alcohol. The CAGE questionnaire can be reliably used to assess alcohol use disorders in schizophrenic patients: with a cutoff score of 1 or more, the sensitivity of the CAGE questionnaire was 0.91 and the specificity was 0.83. With a cutoff score of 2 or more, the sensitivity of the CAGE questionnaire was 0.82 and the specificity was 0.94. The post test probabilities (PPV) were high (PPV1=0.74; PPV2=0.85) compared to the primary probability of alcoholism (29.8%). PMID- 16314078 TI - Evaluation of hydrodynamics in the basket dissolution apparatus using computational fluid dynamics--dissolution rate implications. AB - The aim of this work was to simulate the fluid flow in the basket dissolution apparatus using computational fluid dynamics (CFD) and to use the resulting velocity data (in combination with velocity data from simulated flow fields of the paddle dissolution apparatus) to relate velocities in the vicinity of a dissolving surface to dissolution rate. A further objective of the work was to compare fluid velocities between the basket and paddle dissolution apparatuses. CFD simulations of the basket apparatus were carried out using Fluent software. Flow field solutions were compared with results from flow visualisation techniques and with published ultrasound-pulse-echo velocity data. Velocity data from the flow field solution revealed velocities within the basket to be of the same order as those at the base of the paddle apparatus at the same rotation speed, supporting equivalent dissolution rate data from these locations. Dissolution rates were obtained for compacts of benzoic acid dissolved in 0.1 M HCl at 37 degrees C in the basket apparatus at 50 rpm. The relationship between maximum velocity in the vicinity of a dissolving surface and dissolution rate data from both the paddle and basket apparatuses was in good agreement with theoretical predictions. Analysis of the dissolution rates suggests a significant contribution from free convection in regions of low velocity at the base of the vessel of the basket apparatus. PMID- 16314079 TI - Preparation and release of salbutamol from chitosan and chitosan co-spray dried compacts and multiparticulates. AB - Chitosan microparticulates were prepared by spray drying from aqueous media containing hydrochloric acid or acetic acid. The medium affected the morphology and degree of acetylation of chitosan, the presence of acetic acid resulting in increased acetylation of the polymer during processing. Co-spray drying salbutamol sulphate/chitosan systems with the crosslinking agent formaldehyde had no detectable effect on particle morphology. However, with increasing salbutamol loading particles became less spherical, taking on a collapsed appearance. Spray dried chitosan-salbutamol sulphate microparticulates were X-ray amorphous. Chitosan-salbutamol sulphate composites were compressed into discs to quantify drug release and showed delayed release of salbutamol sulphate. The general power law equation fitted the data better than the t0.5, mono- or bi-exponential models and gave n indices greater than 0.5, i.e. in the range 0.53-0.71. Crosslinking did not dramatically alter the drug release behaviour. Both crosslinked and non crosslinked composites swelled during release, the former to the greater extent. The release data for crosslinked composites gave slightly higher n values than the corresponding non-crosslinked composites, consistent with the increased swelling of these systems. Release studies were also conducted on the microparticulates. Because of the small particle size and large surface area present, the release of the highly soluble drug salt was extremely rapid (> 90% release in 5 min). Twin impinger analysis indicated good in vitro deposition of the microparticulates and potential for pulmonary delivery. PMID- 16314080 TI - A novel large-scale production system for modified basement membrane matrices using gene-swapped parietal endoderm cells. AB - Parietal endoderm-like cells, including Engelbreth-Holm-Swarm tumor and differentiated F9 embryonal carcinoma cells, produce huge amounts of basement membrane components, including laminin-1 (alpha1beta1gamma1). We employed a double-lox system-based gene-swapping strategy in F9 cells to replace the laminin alpha1 gene with a laminin alpha5 minigene. The gene-swapped F9 cells secreted laminin-10 (alpha5beta1gamma1) consisting of the exogenous alpha5 subunit and endogenous beta1 and gamma1 subunits on differentiation. The laminin-10 concentration in the conditioned medium exceeded 10 mg/l, which is 10-fold higher than the concentrations achieved by conventional recombinant expression systems. The gene-swapped F9 cells deposited basement membrane-like matrices containing laminin-10 on culture dishes, offering a novel microenvironment for in vitro cell manipulation. PMID- 16314081 TI - Abnormality of erythrocyte membrane n-3 long chain polyunsaturated fatty acids in sickle cell haemoglobin C (HbSC) disease is not as remarkable as in sickle cell anaemia (HbSS). AB - Sickle cell disease (SCD) is a group of inherited blood disorders in which clinical illness results from the presence of erythrocytes with sickled haemoglobin (HbS). Blood vessel occlusion is a fundamental pathological process in SCD. Sickle cell haemoglobin C (HbSC) disease and sickle cell anaemia (HbSS) share some pathophysiology and clinical manifestations. However, the former is generally less severe. Erythrocytes of HbSC patients have longer life span, reduced haemolysis, and lower propensity to adhere to vascular endothelium than those of their HbSS counterparts. The structure and function of erythrocytes are strongly modulated by membrane long chain polyunsaturated fatty acids (LCPUFA). We have tested the possibility that HbSC and HbSS patients have different membrane fatty acid composition consistent with the difference in their clinical severity. Steady-state patients, 9 HbSC and 28 HbSS, and 15 HbAA were studied. The HbSC patients had a higher level of linoleic (LA, P<0.05) and docosahexaenoic (DHA, P<0.05) acids, and lower arachidonic acid (AA, P<0.01) and AA/eicosapentaenoic acid (EPA) ratio (P<0.05) in erythrocyte choline phosphoglycerides (CPG) compared with the HbSS group. Similarly, the level of EPA was higher and AA/EPA ratio (P<0.01) lower in serine phosphoglycerides of the HbSC patients. In contrast to the HbSC, the HbSS group had lower levels of EPA (P<0.001), DHA (P<0.05), total n-3 metabolites and total n-3 fatty acids (P<0.001) in erythrocyte CPG compared with the healthy HbAA controls. Moreover, the HbSS patients with disease complications compared with those without complications had reduced DHA and total n-3 fatty acids (P<0.005) in erythrocyte CPG. The abnormalities in erythrocyte in LCPUFA which is manifested by an increase in AA and a decrease in EPA and DHA in HbSS relative to HbSC disease observed in this study are consistent with the contrast in clinical severity between the two entities. PMID- 16314082 TI - Supplementation with flax oil and vitamin C improves the outcome of Attention Deficit Hyperactivity Disorder (ADHD). AB - Considerable clinical and experimental evidence now supports the idea that deficiencies or imbalances in certain highly unsaturated fatty acids may contribute to a range of common developmental disorders including Attention Deficit Hyperactivity Disorder (ADHD). Few intervention studies with LCPUFA supplementation have reported inconsistent and marginal results. This pilot study evaluates the effect of alpha linolenic acid (ALA)-rich nutritional supplementation in the form of flax oil and antioxidant emulsion on blood fatty acids composition and behavior in children with ADHD. Post-supplementation levels of RBC membrane fatty acids were significantly higher than pretreatment levels as well as the levels in control. There was significant improvement in the symptoms of ADHD reflected by reduction in total hyperactivity scores of ADHD children derived from ADHD rating scale. PMID- 16314083 TI - Intrabreath analysis of carbon monoxide uptake during exercise in patients at risk for lung injury. AB - The single exhalation analysis of carbon monoxide, acetylene, and methane allows the determination of intrabreath (regional) DL, pulmonary capillary blood flow and ventilation inhomogeneities during rest and exercise. We reasoned that this technique might be more sensitive in detecting regional pulmonary capillary abnormalities than resting single breath DL (DL(sb)). We selected a group of breast cancer patients in high-dose chemotherapy (HDCT) protocols who were at risk for pulmonary injury. We grouped the patients into pre-HDCT and post-HDCT, and used resting DL(sb) to further categorize the latter into those with and without pulmonary injury. We found that exercise DL increases were blunted in post-HDCT patients with low resting DL(sb). More importantly, even in post-HDCT patients with normal resting DL(sb), exercise DL response was reduced in the slowest emptying lung units along with evidence for ventilation inhomogeneities (increased methane slope). We conclude that exercise assessments of DL at low lung volumes and gas mixing properties may be sensitive indicators of lung injury. PMID- 16314084 TI - Phytotoxicity of surface waters of the Thames and Brisbane River estuaries: a combined chemical analysis and bioassay approach for the comparison of two systems. AB - The Thames Estuary, UK, and the Brisbane River, Australia, are comparable in size and catchment area. Both are representative of the large and growing number of the world's estuaries associated with major cities. Principle differences between the two systems relate to climate and human population pressures. In order to assess the potential phytotoxic impact of herbicide residues in the estuaries, surface waters were analysed with a PAM fluorometry-based bioassay that employs the photosynthetic efficiency (photosystem II quantum yield) of laboratory cultured microalgae, as an endpoint measure of phytotoxicity. In addition, surface waters were chemically analysed for a limited number of herbicides. Diuron, atrazine and simazine were detected in both systems at comparable concentrations. In contrast, bioassay results revealed that whilst detected herbicides accounted for the observed phytotoxicity of Brisbane River extracts with great accuracy, they consistently explained only around 50% of the phytotoxicity induced by Thames Estuary extracts. Unaccounted for phytotoxicity in Thames surface waters is indicative of unidentified phytotoxins. The greatest phytotoxic response was measured at Charing Cross, Thames Estuary, and corresponded to a diuron equivalent concentration of 180 ng L(-1). The study employs relative potencies (REP) of PSII impacting herbicides and demonstrates that chemical analysis alone is prone to omission of valuable information. Results of the study provide support for the incorporation of bioassays into routine monitoring programs where bioassay data may be used to predict and verify chemical contamination data, alert to unidentified compounds and provide the user with information regarding cumulative toxicity of complex mixtures. PMID- 16314085 TI - Prevention and treatment of cancer cachexia: new insights into an old problem. AB - Cancer cachexia (CC) is a multifactorial paraneoplastic syndrome characterized by anorexia, body weight loss, loss of adipose tissue and skeletal muscle, accounting for at least 20% of deaths in neoplastic patients. CC significantly impairs quality of life and response to anti-neoplastic therapies, increasing morbidity and mortality of cancer patients. Muscle wasting is the most important phenotypic feature of CC and the principal cause of function impairment, fatigue and respiratory complications, mainly related to a hyperactivation of muscle proteolytic pathways. Most therapeutic strategies to CC have proven to be only partially effective . The inhibition of catabolic processes in muscle has been attempted pharmacologically with encouraging results in animal models. However, data in the clinical setting are still scanty and contradictory. Stimulation of muscle anabolism could represent a promising and valid therapeutic alternative for cancer-related muscle wasting. This goal may be currently achieved with the conventional, short-acting and adverse side effect-rich anabolic steroids. Insulin-like growth factor-1 (IGF-1) plays a critical role in muscle homeostasis, hypertrophy and regeneration. IGF-1 overexpression at the muscular level by gene therapy reverses muscle hypotrophy secondary to catabolic conditions and induces muscle hypertrophy increasing muscle mass and strength. This allows the speculation that this approach could also prove effective in modulating cancer induced muscle wasting, while avoiding the potentially hazardous side effects of systemic IGF-1 administration. The present review will focus on the potential biochemical and molecular targets of CC therapy, and will define the rationale for a novel, gene therapy-based approach. PMID- 16314086 TI - Isolated loco-regional recurrence of breast cancer is more common in young patients and following breast conserving therapy: long-term results of European Organisation for Research and Treatment of Cancer studies. AB - The aim of this study was to evaluate prognostic factors for isolated loco regional recurrence in patients treated for invasive stage I or II breast cancer. The study population comprised 3602 women who had undergone primary surgery for early stage breast cancer, who were enrolled in European Organisation for Research and Treatment of Cancer (EORTC) trials 10801, 10854, or 10902, by breast conservation (55%) and mastectomy (45%). The median follow-up time varied from 5.3 (range: 0.6-9.5) to 11.9 years (range: 0.6-17.4). Main outcome was the occurrence of isolated loco-regional recurrence. The results of multivariate analysis showed that younger age and breast conservation were risk factors for isolated loco-regional recurrence (breast cancer under 35 years of age versus over 50 years of age: hazard ratio 2.80 (95% CI 1.41-5.60)); breast cancer age 35 50 years versus over 50 years: hazard ratio 1.72 (95% CI 1.17-2.54); breast conservation (hazard ratio: 1.82 (95% CI 1.17-2.86)). After perioperative chemotherapy, less isolated loco-regional recurrences were observed (hazard ratio 0.63 (95% CI 0.44-0.91)). No significant interaction effects were observed. It is concluded that young age and breast conserving therapy are both independent predictors for isolated loco-regional recurrence. As an isolated loco-regional recurrence is a potentially curable condition, women treated with breast conservation or diagnosed with breast cancer at a young age should be monitored closely to detect local recurrence at an early stage. PMID- 16314087 TI - Clinical factors associated with fatigue in haematologic cancer patients receiving stem-cell transplantation. AB - We have evaluated risk factors associated with fatigue in 220 cancer patients during hospitalization for stem-cell transplantation (SCT). Fatigue was assessed using a validated one-item energy scale and a comprehensive set of fatigue predictors, at hospital admission (baseline), day of SCT, and 7 days and 14 days after SCT. In cross-sectional multivariate analysis, depression was the variable most consistently and strongly associated with fatigue; other factors significantly associated with fatigue at some time during the study included older age, higher education, smoking, lower Karnofsky performance status, loss of appetite, nausea/vomiting, pain, higher regimen-related toxicity, low hemoglobin level, requirement for red blood-cell transfusions, and third year of the study period. In prospective multivariate analysis, baseline depression showed significance or a trend towards significance in its ability to predict subsequent measures of fatigue during hospitalization. Our findings may help to shed light on the mechanisms underlying fatigue and may also guide future interventions. PMID- 16314088 TI - Glutathione S-transferase M1 and T1 genetic polymorphisms are not related to the risk of hepatocellular carcinoma: a study in the Spanish population. AB - Glutathione S-transferases constitute a superfamily of enzymes that catalyse the inactivating conjugation of endogenous and environmental substrates involved in the pathogenesis of hepatocellular carcinoma (HCC) and glutathione. Genes encoding either glutathione S-transferase Mu-1 or Theta-1 (GSTM1 and GSTT1, respectively) isoforms are polymorphic. Homozygotes for the mutated inactive alleles of each gene are devoid of any specific enzymatic activity (null genotypes). Our aim was to investigate whether individuals with null GST genotypes have a higher risk of developing HCC. A total of 184 Caucasian Spanish patients with a diagnosis of HCC and 329 healthy controls of the same ethnic origin were included. Polymorphisms in GSTM1 and GSTT1 genes were identified through multiplex polymerase chain reactions, and the dihydrofolate reductase (DHFR) gene was used as internal control. No differences were found between the frequencies of GSTM1 (47.8% versus 45.3%) and GSTT1 (28.8% versus 23.1%) null genotypes in cases and controls, respectively, nor in the proportion of carriers of two, one or no active genotypes. Gender, age at diagnosis, tobacco use, chronic infection with hepatitis B or C virus and alcohol abuse did not influence these results. In conclusion, polymorphisms in GSTM1 and GSTT1 genes are not related to the incidence of HCC in a high-risk Spanish population. PMID- 16314089 TI - Optimization of anaerobically digested distillery molasses spent wash decolorization using soil as inoculum in the absence of additional carbon and nitrogen source. AB - The aim of this study was to achieve maximum decolorization of molasses spent wash (MSW) in absence of any additional carbon or nitrogen source using soil as inoculum. Soil samples were collected from the MSW disposal site. Colored soil samples exhibited higher pH, sugar and protein as compare to less colored samples. A decolorization of 69% was obtained using 10% (w/v) soil and 12.5% (v/v) MSW after 7 days incubation. Optimized parameters including days--6 days, pH--6, MSW--12.5% and soil concentration--40%, were obtained for maximum decolorization. A decolorization of 81% was achieved using 10% soil and 12.5% MSW after 18 days incubation in absence of any media supplement. Nearly 12% reduction in decolorization activity of the soil sample was observed over a period of 12 months when stored at 6 degrees C. It could be concluded that the decolorization of MSW might be achieved using soil as inoculum without addition of chemical amendments. PMID- 16314090 TI - Effects of Dysoxylum malabaricum Bedd. (Meliaceae) extract on the malarial vector Anopheles stephensi Liston (Diptera: Culicidae). AB - In recent years, use of environmentally friendly and biodegradable natural insecticides of plant origin have received renewed attention as agents for disease vector control. Methanol extracts of leaves from the Indian white cedar Dysoxylum malabaricum Bedd. (Meliaceae) were tested against mature and immature Anopheles stephensi Liston (Diptera) mosquitoes under laboratory conditions. The extract showed strong larvicidal, pupicidal, adulticidal, and antiovipositional activity. The maximum leaf extract concentration tested in this study was 4%, which produced pronounced effects. In general, first and second instars were more susceptible to leaf extract than older insects. Clear dose-response relationships were established, with the highest dose of 4% plant extract causing 97% mortality of first instars. PMID- 16314091 TI - Removal of hexavalent chromium using a novel cross linked xanthated chitosan. AB - Suitability of a novel cross linked, chemically modified chitosan as highly efficient adsorbent for the recovery of toxic chromium(VI) was studied. After cross linking with glutaraldehyde, xanthate group was grafted onto the back bone of chitosan. Sorption was found to be both pH and concentration dependent, with pH 3 being the optimum value. Both, chemically modified beads (CMCB) and flakes (CMCF) followed a pseudo-second-order kinetics with a rate constant of 2.037 and 4.639 g/mg/min, respectively. The equilibrium data followed the Langmuir isotherm model with maximum capacities of 625 mg/g and 256.4 mg/g and for CMCF and CMCB respectively. Desorption studies revealed the reusability of the sorbent for at least 10 cycles without any significant change in adsorption capacities. PMID- 16314092 TI - Mesophilic anaerobic treatment of sludge from salmon smolt hatching. AB - The mesophilic anaerobic treatment of concentrated sludge from an Atlantic salmon smolt hatchery (total solids (TS): 6.3-12.3wt%) was investigated in a continuous stirred tank reactor (CSTR) at 35 degrees C and 55-60 days hydraulic retention time (HRT). COD-stabilization between 44% and 54% and methane yields between 0.140 and 0.154l/g COD added (0.260-0.281l/g VS added) were achieved. The process was strongly inhibited, with volatile fatty acid concentrations of up to 28 g/l. But the buffer capacity was sufficient to keep the pH-value at 7.4-7.55 during the whole operation. The fertilizing value of the treated sludge was estimated to be 3.4-6.8 kg N and 1.2-2.4 kg P per ton. However, the high VFA content would necessitate special means of application. The energy from the methane that was achieved in the present study would be sufficient to cover about 2-4% of the energy demands of a flow-through hatchery. PMID- 16314093 TI - Separation of proteases from yellowfin tuna spleen by ultrafiltration. AB - Separation of protease, trypsin and chymotrypsin from yellowfin tuna spleen extract by ultrafiltration (UF) using regenerated cellulose membranes with molecular weight cut off (MWCO) 30 and 100 kDa was studied. The 100 kDa membrane had a higher transmission of enzymes than that of the 30 kDa membrane. The enzyme transmission varied from 0.01 to 0.18 and from 0.6 to 0.8 for the 30 kDa membrane and 100 kDa membrane, respectively. The protein transmission was about 0.8 for both membranes. Increasing cross-flow rate and transmembrane pressure (TMP) increased permeate flux. The limiting fluxes at cross-flow rate 120, 240 and 360 L/h for the 30 kDa membrane were 17.3, 43.9 and 54.7 L/m2h, respectively and the limiting fluxes at the same flow rate for 100 kDa membrane were 34.1, 51.1 and 68.4 L/m2h, respectively. The separation of these proteases was achieved using the 30 kDa membrane. The purities of proteases were increased more than ten times at TMP 1.5 bar and cross-flow rate 360 L/h by diafiltration using 30 kDa membrane. PMID- 16314094 TI - Comparison of cellulose solubilisation rates in rumen and landfill leachate inoculated reactors. AB - The aim of this study was to conduct a number of controlled digestions to obtain easily comparable cellulose solubilisation rates and to compare these rates to those found in the literature to see which operational differences were significant in affecting cellulose degradation during anaerobic digestion. The results suggested that differences in volumetric cellulose solubilisation rates were not indicative of the true performance of cellulose digestion systems. When cellulose solubilisation rates were normalised by the mass of cellulose in the reactor at each time step, the comparison of the rates became more meaningful. Cellulose solubilisation was surface area limited. Therefore, changes in the loading rate of cellulose to the reactor altered the volumetric solubilisation rate without changing the mass normalised rate. Comparison of mass normalised solubilisation rates from this study and the literature demonstrated that differences in reactor configuration and operational conditions did not significantly impact on the solubilisation rate whereas the difference in composition of the microbial communities showed a marked effect. This work highlights the importance of using appropriately normalised data when making comparisons between systems with differing operational conditions. PMID- 16314095 TI - The synthesis and aqueous superoxide anion scavenging of water-dispersible lutein esters. AB - Xanthophyll carotenoids of the C40 series, which includes commercially important compounds such as lutein, zeaxanthin, and astaxanthin, have poor aqueous solubility in the native state. Hawaii Biotech, Inc. (HBI) and others have shown that the aqueous dispersibility of derivatized carotenoids can be increased by varying the chemical structure of the esterified moieties. In the current study, the published series of novel, highly water-dispersible C40 carotenoid derivatives has been extended to include (3R,3'R,6'R)-lutein (beta,epsilon carotene-3,3'-diol) derivatives. Two novel derivatives were synthesized by esterification with inorganic phosphate and succinic acid, respectively, and subsequently converted to the sodium salts. Red-orange, clear, aqueous suspensions were obtained after addition of these novel derivatives to USP purified water. Aqueous dispersibility of the disuccinate sodium salt of lutein was 2.85 mg/mL; the diphosphate salt demonstrated a >10-fold increase in dispersibility at 29.27 mg/mL. As reported previously, these aqueous suspensions were obtained without the addition of heat, detergents, co-solvents, or other additives. The direct aqueous superoxide scavenging abilities of these novel derivatives were subsequently evaluated by electron paramagnetic resonance (EPR) spectroscopy in a well-characterized in vitro isolated human neutrophil assay. The novel derivatives were nearly identical aqueous-phase scavengers, demonstrating dose-dependent suppression of the superoxide anion signal (as detected by spin-trap adducts of DEPMPO) in the millimolar range. These lutein based soft drugs will likely find utility in those commercial and clinical applications for which aqueous-phase singlet oxygen quenching and direct radical scavenging may be required. PMID- 16314096 TI - Hydrolysis of plasmid DNA and RNA by amino alkyl naphthalimide as metal-free artificial nuclease. AB - A strategy of dimethylamino alkyldiimide conjugated with an intercalator of naphthalimide for hydrolysis of DNA was suggested and evaluated. 4 can hydrolyze 4 kb plasmid DNA into 2 kb fragments with GC and GG selectivity, which represents a novel example of sequence- or site-selective metal-free DNA artificial nuclease. Results also show it could hydrolyze RNA efficiently. PMID- 16314097 TI - Designing rapid onset selective serotonin re-uptake inhibitors. Part 1: Structure activity relationships of substituted (1S,4S)-4-(3,4-dichlorophenyl)-N-methyl 1,2,3,4-tetrahydro-1-naphthaleneamine. AB - A series of sertraline analogues 4-39 which possess polar groups on the fused tetrahydronaphthalene ring, targeting reduced V(d) as a strategy to reduce T(max) and increase rate of elevation of central 5-HT levels, were prepared. These studies led to the successful identification of 22a, which demonstrated equivalent pharmacology and metabolic stability to 1, but which possessed greatly reduced V(d) leading to significantly shorter T(max), in rat pharmacokinetic studies. PMID- 16314098 TI - 4-Methylideneisoxazolidin-5-ones--a new class of alpha-methylidene-gamma-lactones with high cytostatic activity. AB - A novel, general method of synthesis of 4-methylideneisoxazolidin-5-ones 10 is described. The target compounds were synthesized starting from ethyl 2 diethoxyphosphoryl-2-alkenoates 6 or dicyclohexylammonium 4-diethoxyphosphoryl-2 alkenoates 7. Addition of N-methylhydroxylamine hydrochloride to these Michael acceptors, lactonization to 4-diethoxyphosphorylisoxazolidin-5-ones 9, and Horner Wadsworth-Emmons olefination of formaldehyde using 9 gave the title isoxazolidinones 10. All obtained compounds were tested against L-1210, HL-60, and NALM-6 leukemia cell lines. Several isoxazolidinones 10 were found to be very potent with IC(50)<1 microM. The highest cytostatic activity against HL-60 was observed for 10a and against NALM-6 for 10b with IC(50) values of 0.74 and 0.34 microM, respectively. PMID- 16314099 TI - Changes in 3D joint dynamics during the first 5 months after the onset of independent walking: a longitudinal follow-up study. AB - A longitudinal follow-up study of 10 normally developing children was performed in order to identify changes in mechanical control of gait during the first months after initiation of independent walking. Changes in spatio-temporal parameters and kinematics were recorded (336 trials spread over 83 recording sessions) and linked to kinetic features of gait. At the onset of independent walking, all children in our study group showed the same walking strategy: a dominance of the extending moments around the lower extremity joints was observed and could be linked to the flexed position of the hip and knee during stance. In a subset of our study population, the dominance of the extending moments disappeared with increasing walking experience, though reversal to immature patterns was frequently observed. A linear mixed model showed that with increasing walking experience, there was an increase in dimensionless walking speed, dimensionless cadence and dimensionless stride length (without correction for the increase in speed). Maximal hip extension in stance, knee flexion and ankle plantar flexion at foot contact also increased (even when the increase in speed is taken into account). Dimensionless step width, duty factor, double support time, maximal hip flexion in swing and hip abduction significantly decreased (with correction for speed). Important changes were also observed in ground reaction force patterns, evolving towards a double "hump". No significant changes could be observed in other kinetic parameters, probably due to the high degree of variability. PMID- 16314100 TI - Spatiotemporal characteristics of the walk-to-run and run-to-walk transition when gradually changing speed. AB - The purpose of this study was to examine spatiotemporal parameters of the walk-to run transition (WRT) and run-to-walk transition (RWT) when speed is altered with different constant accelerations. Twenty women (height: 168.9+/-3.36cm) performed three accelerations (0.05, 0.07 and 0.1ms(-2)) and three decelerations (-0.05, 0.07 and -0.1ms(-2)) on a motor-driven treadmill. The transition step in the WRT (first step with a flight phase) and RWT (first step with a double stance phase) occurred at the same speed for all accelerations but these did not occur in the same way. The most striking difference was the presence of a transition step with specific spatiotemporal characteristics in the WRT, whereas this was not observed in the RWT. The transition is not a sudden one-step-event. WRT occurred before transition and consisted of a "pre-transition period" and the transition step whereas RWT occurred after transition and consisted of the transition step and a "post-transition period". Both transition periods were characterized by an exponential evolution of step frequency and step length. Step frequency and step length showed a linear evolution before and after transition. The flight phase of the transition step in the WRT reached a minimum with comparable duration of the last flight phase in the RWT. The flight phase could be considered as an intrinsic dynamical factor of transition. Further research in kinematics, the trajectory of the body centre of mass and energy fluctuations will give more insight in these transitions. PMID- 16314101 TI - Synthesis and biological evaluation of B-ring analogues of (-)-rhazinilam. AB - Three macrocyclic analogues of rhazinilam 1 having a 11- or 12-membered B-ring with an endocyclic carbamate group or an amino-acid residue were synthesized from the natural product. These analogues 3 and 4 displayed a very low activity on tubulin. Thirty N-1 and C-16 substituted analogues of rhazinilam were also synthesized regioselectively from rhazinilam. Stereochemical analyses showed that N-1 and C-16alpha analogues have the same conformation as rhazinilam, whereas C 16beta analogues adopt a different conformation for rings B and D. All N-1 and C 16 analogues were less active than rhazinilam on tubulin, though analogues 5a, 6aalpha, 6balpha, and 6f having the less bulky substituents retained close affinities. A few analogues either active (like 6f) or inactive (like 5o) on tubulin showed significant inhibition of the growth of KB cancer cells. PMID- 16314102 TI - Reduction of nonspecific binding proteins to self-assembled monolayer on gold surface. AB - We developed a gold coated glass chip bearing a poly(ethyleneglycol) (PEG) type compound as hydrophilic spacer for surface plasmon resonance studies, which enabled adequate estimation of K(d) value between FK506 and FKBP12 not only using purified FKBP12 (K(d)=22 nM) but also using Escherichia coli lysate expressing FKBP12 (K(d)=15 nM). These results indicated effectiveness of the PEG spacer for reduction of nonspecific interactions. Chemical stability and simple surface structure of the novel chip are also attractive. PMID- 16314103 TI - Synthesis and anti-HIV activity of bi-functional betulinic acid derivatives. AB - Betulinic acid (BA) derivatives with a side chain at C-3 can inhibit HIV-1 maturation. On the other hand, BA derivatives with a side chain at C-28 can block HIV-1 entry. In order to combine the anti-maturation and anti-entry activities in a single molecule, new bi-functional BA derivatives containing side chains at C-3 and C-28 have been synthesized. The most potent compound ([[N-[3beta-O-(3',3' dimethylsuccinyl)-lup-20(29)-en-28-oyl]-7-aminoheptyl]-carbamoyl]methane) inhibited HIV-1 at an EC50 of 0.0026 microM and was at least 20 times more potent than either the anti-maturation lead compound DSB or the anti-entry lead compound IC9564. This bi-functional BA derivative was active against both HIV entry and maturation. These results suggest that bi-functional BA derivatives with dual mechanisms of action have the potential to become clinically useful for AIDS therapy. PMID- 16314104 TI - 2-Aminothiophene-3-carboxylates and carboxamides as adenosine A1 receptor allosteric enhancers. AB - Three series of 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene and 2-amino-5,6,7,8 tetrahydrocyclohepta[b]thiophenes with 3-carboxylates and carboxamides have been prepared using the Gewald synthesis and evaluated as A(1)AR allosteric enhancers. The structure-activity relationships of these classes of compound are described. A number of compounds, notably 7b, are more potent and efficacious than PD81,723 (1). PMID- 16314105 TI - Diversity-oriented chemical modification of heparin: Identification of charge reduced N-acyl heparin derivatives having increased selectivity for heparin binding proteins. AB - The diversity-oriented chemical modification of heparin is shown to afford charge reduced heparin derivatives that possess increased selectivity for binding heparin-binding proteins. Variable N-desulfonation of heparin was employed to afford heparin fractions possessing varied levels of free amine. These N desulfonated heparin fractions were selectively N-acylated with structurally diverse carboxylic acids using a parallel synthesis protocol to generate a library of 133 heparin-derived structures. Screening library members to compare affinity for heparin-binding proteins revealed unique heparin-derived structures possessing increased affinity and selectivity for individual heparin-binding proteins. Moreover, N-sulfo groups in heparin previously shown to be required for heparin to bind specific proteins have been replaced with structurally diverse non-anionic moieties to afford identification of charge-reduced heparin derivatives that bind these proteins with equivalent or increased affinity compared to unmodified heparin. The methods described here outline a process that we feel will be applicable to the systematic chemical modification of natural polyanionic polysaccharides and the preparation of synthetic oligosaccharides to identify charge-reduced high affinity ligands for heparin-binding proteins. PMID- 16314106 TI - Syntheses and evaluation of fluorinated conformationally restricted analogues of GABA as potential inhibitors of GABA aminotransferase. AB - Inhibition of gamma-aminobutyric acid aminotransferase (GABA-AT) could raise the concentration of GABA, an inhibitory neurotransmitter in the human brain, and could have therapeutic applications for a variety of neurological diseases including epilepsy. Four fluorine-containing analogues of GABA with conformations restricted by a cyclohexane ring system were designed and synthesized, but unlike some of their five-membered ring counterparts, minimal inhibition of GABA-AT was observed. It is likely that the rigid chair conformation of these compounds cannot be accommodated well in the enzyme's active site. PMID- 16314107 TI - Inhibition of cellular proliferation by diterpenes, topoisomerase II inhibitor. AB - We examined the effects of 12 terpene compounds derived from the roots of Euphorbia kansui on the proliferative activity of Xenopus embryo cells. Eight of these compounds showed significant inhibition of cellular proliferation even at low concentrations, while four of them needed to be present at higher concentrations to inhibit cellular proliferation. In order to define the mechanism of inhibition of cellular proliferation by these compounds, the effects of diterpene compounds on the activity of topoisomerase II were measured. Most of the diterpene compounds that inhibited cellular proliferation also inhibited topoisomerase II activity. PMID- 16314108 TI - Phosphoramidate and phosphate prodrugs of (-)-beta-D-(2R,4R)-dioxolane-thymine: synthesis, anti-HIV activity and stability studies. AB - A series of phosphoramidate and phosphate prodrugs of DOT were synthesized via dichlorophosphate or H-phosphonate chemistry and evaluated for their anti-HIV activity against LAI M184V mutants in PBM cells as well as for their cytotoxicity. The antiviral and cytotoxic profiles of the prodrugs were compared with that of the parent compound (DOT), and it was found that four aryl phosphoramidates 5, 18, 20, and 26 showed a significant enhancement (8- to 12 fold) in anti-HIV activity without cytotoxicity. Chemical stability of these prodrugs was evaluated in phosphate buffer at pH values of biological relevance (i.e., pH 2.0 and 7.4). Enzymatic hydrolysis was also studied in esterase or lipase in buffer solution. Chemical stability studies indicate that the phosphoramidates have good chemical stability at pH 2.0 and at pH 7.4 phosphate buffer. Phosphoramidate prodrugs were hydrolyzed in vitro by esterase or lipase and found to be better substrates for lipases than for esterases. 1,3-Diol cyclic phosphates showed potent anti-HIV activity without increasing the cytotoxicity compared with that of DOT and have good chemical and enzymatic stability. Long chain lipid phosphates, although showed potent anti-HIV activity, exhibited increased cytotoxicity. PMID- 16314109 TI - A geographic information systems (GIS) and spatial modeling approach to assessing indoor radon potential at local level. AB - This study integrates residential radon data from previous studies in Southern California (USA), into a geographic information system (GIS) linked with statistical techniques. A difference (p<0.05) is found in the indoor radon in residences grouped by radon-potential zones. Using a novel Monte Carlo approach, we found that the mean distance from elevated-radon residences (concentration>74 Bq m(-3)) to epicenters of large (> 4 Richter) earthquakes was smaller (p<0.0001) than the average residence-to-epicenter distance, suggesting an association between the elevated indoor-radon and seismic activities. PMID- 16314110 TI - A regulated switch of chick neurofascin isoforms modulates ligand recognition and neurite extension. AB - Neural cell adhesion molecule neurofascin regulates the induction of neurite outgrowth, the establishment of synaptic connectivity and myelination. Neurofascin isoforms are generated by spatially and temporally controlled alternative splicing. Isoform NF166 is predominantly expressed in dorsal root ganglia from embryonal day 5 (E5) to E8, and a further neurofascin isoform NF185 appears at E9. Expression of neurofascin and its binding partner axonin-1 on sensory fibers implies functional interactions for neurite outgrowth. E7 sensory neurons require NF166-axonin-1 interactions for neurite extension, accordingly. The contribution of NF166-axonin-1 interaction for neurite outgrowth decreases in parallel with the appearance of NF185 on sensory neurons at E9. This finding may be explained by (1) alleviated intrinsic capability to use axonin-1 as a cellular receptor and (2) reduced binding of axonin-1 to NF185. Finally, NF166, but not NF185, serves as a cellular receptor for neurite induction via homophilic interactions with a neurofascin substrate. PMID- 16314111 TI - Thermodynamic and electrostatic properties of ternary Oxytricha nova TEBP-DNA complex. AB - Telomeres constitute the nucleoprotein ends of eukaryotic chromosomes which are essential for their proper function. Telomere end binding protein (TEBP) from Oxytricha nova was among the first telomeric proteins, which were well characterized biologically. TEBP consists of two protein subunits (alpha, beta) and forms a ternary complex with single stranded telomeric DNA containing tandem repeats TTTTGGGG. This work presents the characterization of the thermodynamic and electrostatic properties of this complex by computational chemistry methods (continuum Poisson-Boltzmann and solvent accessible surface calculations). Our calculations give a new insight into molecular properties of studied system. Based on the thermodynamic analysis we provide a rationale for the experimental observation that alpha and ssDNA forms a binary complex and the beta subunit joins alpha:ssDNA complex only after the latter is formed. Calculations of distribution of the molecular electrostatic potential for protein subunits alone and for all possible binary complexes revealed the important role of the "guiding funnel" potential generated by alpha:ssDNA complex. This potential may help the beta subunit to dock to the already formed alpha:DNA intermediate in highly steric and electrostatic favorable manner. Our pK(a) calculations of TEBP are able to explain the experimental mobility shifts of the complex in electrophoretic non-denaturating gels. PMID- 16314112 TI - The taxonomic status of 'phylogroups' in the Parus teneriffae complex (Aves): comments on the paper by Kvist et al. (2005). PMID- 16314113 TI - Phylogeny of the ectomycorrhizal mushroom genus Alnicola (Basidiomycota, Cortinariaceae) based on rDNA sequences with special emphasis on host specificity and morphological characters. AB - Alnicola (=Naucoria, pro parte) is a mushroom genus of strictly temperate, obligately ectomycorrhizal species, traditionally included in the family Cortinariaceae. Most Alnicola spp. are primarily host specific on Alnus, although a few are mycobionts of Salix or other hosts. The different species of Alnicola exhibit unique morphological (cystidia, pileipellis) and cytological (dikaryotic or monokaryotic hyphae) characters. This makes the genus Alnicola of particular interest for studying the evolution of host specificity and morphological characters in ectomycorrhizal basidiomycetes. We used a combination of classical morphological and phylogenetic methods (rDNA ITS and LSU sequences) to address the following questions: (i) Is Alnicola monophyletic? And (ii) Are characters like host specificity or microscopical structures synapomorphic for certain clades? The study included nearly all currently known European Alnicola sp. Our results demonstrated that, on one hand, the genus Alnicola is polyphyletic, with sistergroup relationships to Hebeloma, Anamika or the clades /Hymenogaster I and /Hymenogaster II. On the other hand, Alnicola splits into three well-supported clades corresponding to the sections Alnicola, Submelinoides, and Salicicolae. The strict host-specificity to Alnus is a derived character and has occurred at least twice. The following morphological characters are synapomorphic for defined clades: the spindle-shaped hymenial cystidia for sect. Alnicola, the hymeniform pileipellis for sect. Submelinoides, and monocaryotic/clampless hyphae for sect. Salicicolae and its sistergroup /Hymenogaster II. As a taxonomical consequence, polyphyly of Alnicola implies that the sects. Submelinoides and Salicicolae need to be segregated from Alnicola. PMID- 16314114 TI - Generating single-copy nuclear gene data for a recent adaptive radiation. AB - Recent adaptive radiations provide an exceptional opportunity to understand the processes of speciation and adaptation. However, reconstructing the phylogenetic history of recent and rapidly evolving clades often requires the use of multiple, independent gene genealogies. Nuclear introns are an obvious source of the necessary data but their use is often limited because degenerate primers can amplify paralogous loci. To identify PCR primers for a large number of loci in an especially rapid adaptive radiation, that of the flowering plant genus Aquilegia (Ranunculaceae), we developed an efficient method for amplifying multiple single copy nuclear loci by sequencing a modest number of clones from a cDNA library and designing PCR primers; with one primer anchored in the 3' untranslated region (3' UTR) and one primer in the coding region of each gene. Variation between paralogous loci evolves more quickly in 3'-UTR regions compared to adjacent exons, and therefore we achieved high specificity for isolating orthologous loci. Furthermore, we were able to identify genes containing large introns by amplifying genes from genomic DNA and comparing the PCR product size to that predicted from their cDNA sequence. In Aquilegia eight out of eleven loci were isolated with this method and six of these loci had introns. Among four genes sequenced for samples spanning the phylogenetic breadth of the genus, we found sequence variation at levels similar to that observed in ITS, further supporting the recent and rapid radiation in Aquilegia. We assessed the orthology of amplification products by phylogenetic congruence among loci, the presence of two well established phylogenetic relationships, and similarity among loci for levels of sequence variation. Higher levels of variation among samples for one locus suggest possible paralogy. Overall, this method provides an efficient means of isolating predominantly single-copy loci from both low and high-copy gene families, providing ample nuclear variation for reconstructing species-level phylogenies in non-model taxa. PMID- 16314116 TI - Molecular phylogeny of the stromateoid fishes (Teleostei: Perciformes) inferred from mitochondrial DNA sequences and compared with morphology-based hypotheses. AB - The phylogenetic relationships among 21 species of stromateoid fishes, representing five families and 13 genera, were reconstructed using 3263bp of mitochondrial DNA sequences, including the posterior half of the 16S rRNA and entire COI and Cytb genes. The resultant molecular phylogenies were compared with previous phylogenetic hypotheses inferred from morphological characters. Molecular phylogenetic trees were constructed using the maximum parsimony, maximum likelihood, and Bayesian methods. All three methods resulted in well resolved trees with most nodes being supported by moderate to high support values. In contrast to previous morphological analyses, which resulted in non monophyly of Centrolophidae, all three methods utilized for the present molecular analyses supported the monophyly of Centrolophidae, as well as the reciprocal monophyly of the other stromateoid families, previous morphological hypotheses being rejected by the Templeton and Shimodaira-Hasegawa tests. In addition, the three methods indicated a sister-group relationship between Ariommatidae and Nomeidae. The position of Tetragonuridae was, however, incongruent between the MP method and the ML and Bayesian methods, being placed in the most basal position of Stromateoidei in the former, but occupying a sister relationship to Stromateidae in the latter. Comparison of the molecular phylogenies to previous morphological hypotheses suggested that evolutionary changes in morphological characters have not occurred equally among the stromateoid lineages, the evolution of the centrolophids not having been accompanied by appreciable morphological changes, whereas other stromateoids have undergone considerable morphological changes during their evolutionary history. The molecular phylogenies also shed some light on the evolutionary pattern of the pharyngeal sac, two of the four types of sac corresponding to two main lineages of Stromateoidei. Some taxonomic implications were also discussed. PMID- 16314117 TI - Reappraisal of the Hansemann-Boveri hypothesis on the origin of tumors. AB - Cancer is now known to be a genetic disease. In tumor development, cell nuclei undergo mutations, which can result in cytologically visible chromosome aberrations. The aneuploid errors may involve amplification or deletion of whole chromosomes or segments thereof. David Hansemann [1858-1920] and Theodor Boveri [1862-1915] were major contributors to early debates on the relationship between chromosomal defects, tumorigenesis and malignancies. In 1890, Hansemann observed asymmetrical nuclear divisions in human epithelial cancers. In these abnormal, but bipolar, divisions, a fraction of the chromosomes fails to segregate properly. Hansemann carefully documented the occurrence of asymmetric divisions in a wide variety of tumors. However, he remained a lifelong skeptic with regard to whether such events could be considered the underlying cause of tumors. Almost a quarter of a century after Hansemann's initial observations, Boveri considered the origin of tumors based on his earlier recognition of the functional specificity of each chromosome. He also explicitly drew on Hansemann's observations in proposing a model for tumorigenesis. Its central tenet was that a tumor typically originates from a single cell that has inherited a defined, but incorrectly combined, set of chromosomes. The rare occurrence of a pluripolar spindle represented Boveri's paradigm for a type of abnormal mitosis that can produce a host of random chromosomal combinations. He suggested that some of these combinations will induce tumorous transformation, and will inevitably arise occasionally. Since pluripolar and unbalanced bipolar divisions fail to distribute the hereditary chromatic material correctly, both of these mechanisms can give rise to tumor progenitors. PMID- 16314118 TI - Endopolyploidy as a morphogenetic factor of development. AB - This paper summarizes the works published by author and his co-workers in the Russian journal Tsitologiya concerning endopolyploidy in mollusks and appraises this phenomenon in general. Both ontogenetic and phylogenetic aspects of endopolyploidy have been studied. In the snail Succinea lauta, a complex examination of endomitosis has been performed. A regular replacement of the normal (complete) proliferative mitosis by abnormal (incomplete) restitutional mitosis, and then by Geitler's classic endomitosis has been demonstrated. We examined 29 bivalve and 82 gastropod species for the presence of polyploid cells in glandular tissues and ganglia. In the bivalve species, ordinary diploid cells form various tissues, while in the gastropods, the role of polyploidy in tissue development appears to increase in phylogenesis. The rise of endopolyploidy and cell giantism in histogeneses of a variety of animal and plant species is widely known. It is believed to be a regular event in the evolution of certain groups. To give a universal interpretation of endopolyploidy, we proposed that a single polyploid cell be better considered as an endoclone. In this case, evolutionary transformation of diploid cell clones into polyploid endoclones may be viewed as Dogel's oligomerization applied to cell-tissue level. From this viewpoint, major properties of an oligomerized system (intensification of function, functional efficiency (ergonomy), increased genomes reliability, simplification of the intra and supersystem regulations, and acceleration of development) can be considered as principal peculiarities of polyploid growth strategy. The above peculiarities allow one to consider endopolyploidy as an additional means of integrative onto(histo)genetic regulations and correlations and as an important evolutionary factor (coordinations) acting through natural selection. Thus, in general, endopolyploidy is an adaptive morphogenetic factor, but its concrete role may differ in different tissues and organisms depending on cell specialization and histogenetic particularities. PMID- 16314119 TI - Segregation of genomes in polyploid tumour cells following mitotic catastrophe. AB - Following irradiation p53-function-deficient tumour cells undergo mitotic catastrophe and form endopolyploid cells. A small proportion of these segregates nuclei, and give rise to viable descendants. Here we studied this process in five tumour cell lines. After mitotic failure, tumour cells enter the endocycle and form mono-nucleated or multi-nucleated giant cells (MOGC and MNGC). MNGC arise from arrested anaphases, MOGC, from arrested metaphases. In both cases the individual genomes establish a radial pattern by links to a single microtubule organizing centre. Segregation of genomes is also ordered. MNGC present features of mitosis being resumed from late anaphase. In MOGC the sub-nuclei retain arrangement of stacked metaphase plates and are separated by folds of the nuclear envelope. Mitosis then resumes in sub-nuclei directly from metaphase. The data presented indicate that endopolyploid tumour cells preserve the integrity of individual genomes and can potentially re-initiate mitosis from the point at which it was interrupted. PMID- 16314120 TI - Nuclear deformation characterizes Werner syndrome cells. AB - Mutations in the lamin A gene have been shown, among other defects, to give rise to Hutchinson-Gilford progeria syndrome (HGPS) and to atypical Werner syndrome (WS), both of which are progeroid disorders. Here, we have investigated well characterized WS patient cell strains that are compound heterozygous for mutations in the WRN gene. As in HGPS and in atypical WS, we found nuclear deformations to be characteristic of all cell strains studied. In WS cells centrosome number, assembly of the nuclear lamina and nuclear pore distribution occurred normally. Furthermore, nuclear deformations were not associated with a defect in lamin A expression. We propose that nuclear deformation is a universal characteristic of progeroid cells and may result from slow cell cycle progression. PMID- 16314121 TI - COMBO-FISH for focussed fluorescence labelling of gene domains: 3D-analysis of the genome architecture of abl and bcr in human blood cells. AB - Structural analysis and nanosizing of gene domains requires not only high resolution microscopy but also improved techniques of fluorescence labelling strongly focussed on the gene domains. To investigate the architecture of abl and bcr in blood cell nuclei forming the Philadelphia chromosome in CML, we applied COMBO-FISH using specifically colocalising combinations of triple strand forming oligonucleotide probes for abl on chromosome 9 and bcr on chromosome 22. Each probe set consisting of 31 homopyrimidine oligonucleotides was computer selected from the human genome database. Measurements by 3D microscopy were compared to results obtained after standard FISH using commercially available abl/bcr BAC probes. The relative radial fluorescence distributions in lymphocyte cell nuclei of healthy donors in comparison to cell nuclei of blood cells of CML patients showed a strong correlation in the location of abl and bcr for both labelling techniques. The absolute distances of the homologous bcr domains and the abl domain-nuclear center-abl domain angles in cell nuclei of CML donors differed significantly from those of healthy donors only when COMBO-FISH was applied. These results indicate that COMBO-FISH may be more sensitive than standard FISH in case of slight modifications in the genome architecture. PMID- 16314122 TI - Possible role of endogenous growth inhibitors in regeneration of organs: searching for new approaches. AB - A hydrophilic protein component (12-17 kDa) of the thermostable protein complex (TSPC) of different organs (heart, liver, kidney, brain) of adult white rats has been identified using hydrophobic interaction chromatography (HIC). Chromatograms and spectra of the myocardial hydrophilic component of different animals (snail, pigeon, rat, pig) detected within the UV region (190-360 nm) have shown that in different organs of the same animal species as well as in the same organ (e.g. heart) of different species, TSPC contains identical hydrophilic components, i.e. there is clearly a phylogenetically conserved group of thermostable proteins that regulates proliferation processes. PMID- 16314123 TI - Death of mitochondria during programmed cell death of leaf mesophyll cells. AB - The role of plant mitochondria in the programmed cell death (PCD) is widely discussed. However, spectrum and sequence of mitochondrial structural changes during different types of PCD in leaves are poorly described. Pea, cucumber and rye plants were grown under controlled growing conditions. A part of them were sprinkled with ethylene releaser to accelerate cell death. During yellowing the palisade parenchyma mitochondria were attracted to nuclear envelope. Mitochondrial matrix became electron translucent. Mitochondria entered vacuole by invagination of tonoplast and formed multivesicular bodies. Ethephon treatment increased the frequency of sticking of mitochondria to the nuclear envelope or chloroplasts and peroxisomes. Mitochondria divided by different mechanisms and became enclosed in Golgi and ER derived authopagic vacuoles or in the central vacuole. Several fold increase of the diameter of cristae became typical. In all cases mitochondria were attached to nuclear envelope. It can be considered as structural mechanism of promoting of PCD. PMID- 16314124 TI - Whole-genome chromosome distribution during nuclear fragmentation of giant trophoblast cells of Microtus rossiaemeridionalis studied with the use of gonosomal chromatin arrangement. AB - Gonosomal chromatin bodies (GCBs), i.e. blocks of condensed chromatin consisting of heterochromatized region of the sex chromosomes of the field vole M. rossiaemeridionalis, were used as a natural interphase chromosome marker in order to clarify the regularities of GCB rearrangement during nuclear fragmentation of secondary giant trophoblast cells (SGTCs) at the end of their differentiation. Cytophotometrical measurements of DNA content in the nuclei, nuclear fragments and simultaneously in the GCBs were made in the secondary giant SGTCs of field vole M. rossiaemeridionalis. In most cases 1 to 2 GCBs get into the nuclear fragments at different ploidy levels. In the nuclear fragments, GCB DNA content decreased mostly proportionally to DNA content in the whole fragments corresponding to 2c, 4c and 8c. The data obtained demonstrate a regular whole genome chromosome distribution into nuclear fragments. A possible mechanism of nuclear fragmentation that largely ensures a balanced genome in nuclear fragments is discussed. PMID- 16314125 TI - Nucleolar localization of Aspergillus fumigatus CgrA is temperature-dependent. AB - Pathogenic fungi must adapt to multiple adverse environmental conditions during the transition from the environment to a mammalian host, one of which is temperature. The ability of Aspergillus fumigatus to grow optimally under conditions of thermal stress requires the nucleolar protein CgrA. In this study, we have determined how temperature affects the intracellular localization of CgrA in A. fumigatus using a green fluorescent protein (GFP)-tagging approach. At 22 degrees C, CgrA was almost exclusively in the nucleolus, with a ratio of nucleolar to cytoplasmic fluorescence of 10:1. At 37 degrees C, the ratio of nucleolar to cytoplasmic fluorescence was reduced fivefold, and increased correspondingly in the cytoplasm. This effect was not seen with the nucleolar protein NopA in wild-type A. fumigatus. However, in a DeltacgrA mutant NopA was delocalized from the nucleolus at 37 degrees C but not at 22 degrees C. These results provide evidence for a temperature-dependent mechanism of intracellular localization for CgrA, and suggest that CgrA facilitates nucleolar compartmentalization of NopA at higher temperature. PMID- 16314126 TI - Osteochondrosis and copper: histology of articular cartilage from foals out of copper supplemented and non-supplemented dams. AB - Copper (Cu) supplementation of dams in late gestation may be protective against articular cartilage abnormalities in foals. Articular cartilage was harvested from 22 Thoroughbred foals at 160 days of age, at sites predisposed to osteochondrosis (OC), and examined for evidence of early cartilage abnormalities and established dyschondroplastic (DCP) lesions to determine if there were any significant differences due to mare Cu supplementation by injection during late gestation, or foal liver Cu concentration. Cu supplemented mares received calcium Cu edetate injections in late gestation (250 mg at around 220, 248, 276 and 304 days gestation, then every two weeks until foaling). Foals were euthanased at 160 days of age and articular cartilage was harvested from four defined sites. Samples were examined for histological appearance of chondrocytes after staining with haematoxylin and eosin, and were also stained with toluidine blue to indicate proteoglycan content. Alkaline phosphatase (ALP) activity was detected by histochemistry, and histocytochemical techniques were used to determine the expression of cathepsin B. Cu supplementation of the dam, or liver Cu concentration of the foal at birth or 160 days of age had no statistically significant effect on the frequency of cartilage irregularities observed grossly, or abnormalities detected histologically at four defined sites. ALP expression was similar in all samples. Cathepsin B expression varied between sites, and was seen in chondrocyte clusters. The intensity of toludine blue staining varied between sites. Minor histological cartilage abnormalities were observed in cartilage from clinically normal animals. These abnormalities might be 'early' dyschondroplastic lesions, which could resolve or progress. The role of Cu in the development, resolution or progression of dyschondroplastic lesions is poorly understood. PMID- 16314127 TI - Retrospective epidemiological investigation of an outbreak of bovine ephemeral fever in 1991 affecting dairy cattle herds on the Mediterranean coastal plain. AB - From August to October 1991 bovine ephemeral fever (BEF) occurred sporadically in two localities in Israel. The morbidity and mortality rates reached 2.6% and 0.1%, respectively. Only 12/50 dairy cattle herds were clinically infected with BEF in the dairy community. The total morbidity rate reached 0.8%. The lowest morbidity rate was recorded in young heifers (5.5%) and the highest in adult cows (75%). Only heifers over the age of three months were clinically affected. The spread of the disease apparently followed the local prevailing night winds, which blow from east to west, i.e., from the land toward the sea. The morbidity period lasted 61 days. The low incidence and morbidity rates were possibly due to the low virulence of the virus strain involved in the 1991 epidemic. Retrospective analysis indicates that vectors - apparently mosquitoes - infected with BEF virus could have been overwintering. PMID- 16314128 TI - Cloning and nucleotide sequencing of the second internal transcribed spacer of ribosomal DNA for three species of Eimeria from chickens in Taiwan. AB - Coccidiosis of chickens caused by protozoan parasites of the genus Eimeria (Coccidia: Eimeriidae) is an enteric disease that results in great economic losses throughout the world, including Taiwan. Using polymerase chain reaction (PCR) with primers specific for the second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA), three species of Eimeria, E. tenella, E. maxima, and E. acervulina have been successfully characterised from chickens in Taiwan. The sizes of PCR products from various isolates representing these three species were between 370 and 580 base pairs (bp). After cloning and sequencing of the PCR products, high nucleotide sequence identity (96.8-100%) was observed within a species. In addition, ITS-2 nucleotide sequences for E. tenella had higher homology (98.5-99.3%) than E. maxima (81.6-96.5%) when compared with appropriate sequences deposited in GenBank. To our knowledge, this is the first report of a 412-bp ITS-2 sequence for E. acervulina from chickens. PMID- 16314129 TI - Osteoarthritis: linking nitric oxide and metalloproteinases in canine articular ligament pathology. PMID- 16314130 TI - Effects of cadmium chloride inhalation on airflow limitation to histamine, carbachol and adenosine 5'-monophosphate assessed by barometric whole body plethysmography in healthy dogs. AB - The effects of pharmacological bronchoprovocation on airflow patterns and surrogate respiratory parameters assessed by barometric whole body plethysmography (BWBP) were investigated in healthy dogs, previously exposed to cadmium chloride inhalation. BWBP-derived respiratory variables were calculated (1) at baseline and (2) following nebulisation of increasing concentrations of histamine, carbachol and adenosine 5'-monophosphate (AMP) until enhanced pause (PENH) increased to 300% of baseline (PCPENH300). Bronchoalveolar lavage fluid (BALF) cytology before (BCC) and after (ACC) cadmium chloride inhalation revealed cadmium-induced airway inflammation. Neutrophils increased from 6.7 +/- 7.3% (728 +/- 104/microL) BCC to 77.8 +/- 8.6% (3255 +/- 1407/microL) ACC. PCPENH300 for all three agonists significantly decreased ACC (means+/-SD) as follows: PCPENH300(histamine) 0.72 +/- 0.28 mg/mL BCC, and 0.35 +/- 0.31 mg/mL ACC (P<0.02); PCPENH300(carbachol) 0.34 +/- 0.16 mg/mL BCC, and 0.064 +/- 0.032 mg/mL ACC (P<0.02); PCPENH300(AMP) 1000 mg/mL BCC, and 415 +/- 398 mg/mL ACC (P<0.03). The only clinical sign shown was coughing. It was concluded that airway hyperresponsiveness after induced airway inflammation can be determined by BWBP in conscious small sized dogs. BWBP could be a suitable technique to study the respiratory effects of urban environmental pollution in pets. PMID- 16314131 TI - Ultrasonographic examination of important aspects of the bovine shoulder- physiological findings. AB - The left and right shoulder region of 19 healthy cattle and the left and right shoulders of six bovine cadavers were examined ultrasonographically using a 7.5 MHz linear and a 3.5 MHz convex transducer. The lateral shoulder muscles were successfully imaged in all cases; the joint space appeared as a funnel shaped interruption of the hyperechoic bone surfaces of the distal part of the scapula and the proximal and lateral part of the humeral head. In healthy cattle, the synovial cavities of the scapulohumeral joint, the bicipital bursa and the infraspinous bursa could not be clearly defined. After experimental filling with water, the synovial cavities could be differentiated as discrete anechoic zones. In this study, the normal ultrasonographic appearance of the soft tissue structures and bone surfaces of the scapula and proximal humerus is presented, providing basic reference data for the ultrasonographic evaluation of the bovine shoulder region. PMID- 16314132 TI - Novel polymorphisms at codons 146 and 151 in the prion protein gene of Cyprus goats, and their association with natural scrapie. AB - To discern whether an association exists between specific combinations of polymorphisms of the prion protein (PrP) and natural scrapie in Cyprus goats, 250 goats were examined, including 164 histologically positive cases. Previously reported amino acid polymorphisms were detected at codons 154 (R-->H), 168(P- >Q), 220(Q-->H) and 240 (S-->P) and nucleotide alterations at codons 42 (a-->g) and 138 (c-->t). Additionally, novel amino acid polymorphisms were detected at codons 146 (N-->S or D) and 151 (R-->H) and new "silent" mutations were found at codons 179 (V,g-->t), 181 (D,c-->t) and 219 (T,c-->t). The two novel polymorphisms at codon 146 were found only in the healthy control and scrapie negative goats. By comparison, none of the scrapie-affected goats encoded these polymorphisms. PMID- 16314133 TI - Inter-sexual differences in resting metabolic rates in the Texas tarantula, Aphonopelma anax. AB - Intra-specific variation in life history and mating strategies can lead to differences in energy allocation and expenditure in males and females. This may, in turn, explain large-scale evolutionary patterns. In this study, I investigated the effects of body mass, temperature and sex on resting metabolic rates (RMRs) in sexually mature male and female tarantulas (Aphonopelma anax (Chamberlin)), a species that exhibits extreme inter-sexual differences in life history after reaching sexual maturity. RMRs were measured as rates of CO(2) production in an open-flow respirometry system at 20, 25, 30 and 35 degrees C. These temperatures are typical to what this species experiences under natural conditions. In addition, a respiratory quotient (RQ) of 0.92 was calculated from rates of CO(2) production and O(2) consumption in a closed, constant-volume respirometry system. As expected, RMRs increased with increasing temperature and body mass. However, after adjusting for the influence of body mass, males had substantially higher metabolic rates than females at each temperature. This higher metabolic rate is proposed as an adaptive strategy to support higher energetic demands for males during their active, locomotory search for females during the mating season. PMID- 16314134 TI - Transport of d-galactose by the gastrointestinal tract of the locust, Locusta migratoria. AB - Due to exoskeleton, the absorption of nutrients in adult insects takes place across the gastrointestinal tract epithelium. In most physiological studies, sugar intestinal absorption has been described as a diffusional process and to date no sugar transporter has been cloned from the digestive tract of insects. In the present work, the existence of a saturable transport system for galactose in the gastric caeca of Locusta migratoria is clearly demonstrated. This transport shows a relatively high affinity for galactose (apparent K0.5=2-3 mM) and is inhibited by glucose, 2-deoxyglucose and with less potency by fructose and alpha methyl-d-glucoside. The absence of sodium or the presence of phloridzin hardly affects galactose absorption, indicating that it is not mediated by a SGLT1-like transporter. The absence of K+, Cl-, Mg2+ and Ca2+ or changes in the pH do not modify galactose absorption either. Nevertheless, phloretin, cytochalasin B and theophylline (inhibitors of facilitative transporters) decrease sugar uptake around 50%. Xenopus laevis oocytes microinjected with poly A+ RNA isolated from gastric caeca show sodium-independent galactose uptake that is three times higher than in non-injected oocytes, further supporting the existence of a mRNA coding for at least one equilibrative sugar transporter in L. migratoria gastric caeca. PMID- 16314135 TI - [GnRH analogues and myomas: against but...]. PMID- 16314136 TI - Thymoquinone: a promising anti-cancer drug from natural sources. AB - There has been growing interest in naturally occurring compounds with anti-cancer potential. Black seed is one of the most extensively studied plants. This annual herb grows in countries bordering the Mediterranean Sea and India. Thymoquinone (TQ) is the bioactive constituent of the volatile oil of black seed. It has been shown to exert anti-neoplastic and anti-inflammatory effects. The molecular pathways of TQ action are not clear. Nevertheless, TQ is known to induce apoptosis by p53-dependent and p53-independent pathways in cancer cell lines. Growth inhibition is associated with induction of cell cycle arrest. TQ also acts on the immune system by modulating the levels of inflammatory mediators. To date, the chemotherapeutic potential of TQ in the clinic has not been tested, but numerous studies have shown its promising anti-cancer effects in animal models. The combination of TQ with clinically used anti-cancer drugs has led to improvements in their therapeutic index and prevents non-tumor tissues from sustaining chemotherapy-induced damage. PMID- 16314137 TI - Effects of sucking acidic candies on saliva in unilaterally irradiated pharyngeal cancer patients. AB - Patients who have received radiation therapy on the head and neck area often use acidic candies to relieve symptoms of dry mouth. Therefore, the aim of this study was to determine the erosive potential in relation to teeth of an acidic candy in 10 such patients. The patients sucked the candy while their whole saliva was collected into a closed system at different times: baseline, candy-stimulated, and post-stimulated. The erosive potential of the candy was evaluated from candy induced changes in saliva degree of saturation with respect to hydroxyapatite (HAp). Previously published normative values were used for comparison. The results showed that saliva became significantly more undersaturated with respect to HAp in irradiated patients, and failed to return to baseline values during the post-stimulatory period, which it normally does in healthy individuals. Thus, prevention of dental breakdown in these patients should involve counseling regarding choice of stimulant for dry mouth relief. PMID- 16314138 TI - Evolution of chronic dysphagia following treatment for head and neck cancer. AB - We would like to assess the evolution of chronic dysphagia (1 year or more) following treatment for head and neck cancer. Modified barium swallow (MBS) examinations were performed in cancer-free patients who complained of dysphagia following treatment for head and neck cancer. The severity of the dysphagia was graded on a scale of 1-7. Each patient had at least 2 MBS. Severity of dysphagia was compared between the first and last MBS study to determine whether the swallowing dysfunction had returned to normal. Patients with complaint of dysphagia and normal MBS also underwent a regular barium swallow to assess the structural integrity of the pharynx and esophagus. Between 1996 and 2001, 25 patients with dysphagia underwent repeat MBS following treatment. Swallowing dysfunction did not return to normal in the majority of the patients. At a median time of 26 months following treatment (range 15-82 months), only two patient (8%) had normalization of the swallowing. The severity of dysphagia decreased in eight patients (32%), remained unchanged in 12 patients (48%), and worsened in five patients (20%). Eight patients (32%) still had aspiration problems at 12-83 months following treatment. Six patients (24%) required dilation because of pharyngeal stenosis. Three patients who required dilation had improvement of the dysphagia severity. Chronic dysphagia is a relentless process possibly due to excessive scarring. Patients with chronic dysphagia are at risk of malnutrition, and aspiration. Management of chronic dysphagia requires a team approach with nutritional support, psychological counseling, dilation, and tube feedings when indicated. PMID- 16314139 TI - Genotoxic and mutagenic activity of environmental air samples from different rural, urban and industrial sites in Flanders, Belgium. AB - The present study reports mutagenic and genotoxic activities associated with ambient air collected at 15 sites characteristic for urban, industrial or rural conditions in Flanders. Airborne particulates (PM10) and semi-volatile compounds were collected on quartz filters (QF) and polyurethane foam (PUF) cartridges using a high-volume sampling device. The mutagenic and genotoxic potency of the organic extracts--Soxhlet extraction with acetone--was determined by use of the Salmonella mutagenicity standard plate-incorporation assay and the Vitotox assay, respectively. Concentrations of 16 polycyclic aromatic hydrocarbons (PAHs) in the extracts were determined by reversed-phase high-performance liquid chromatography (HPLC). Ambient air samples contained significant PAH levels and mutagenic activities at all 15 sites: direct mutagenicity of up to 47 revertants per cubic meter was found in the QF extracts and more limited activity of up to 11 rev m( 3) in the PUF extracts. Metabolic activation of PUF extracts resulted in an important increase in mutagenic activity, up to 30 rev m(-3), but no such increase was observed for QF extracts. The highest values were observed outside large cities at industrial sites and at a rural site contaminated by pollution from a chemical plant at a distance of 4 km. Also at the background location near the North Sea a significant mutagenic activity was measured in the QF extracts (+S9: 9 rev m(-3); -S9: 7 rev m(-3)). Apparently, there is in Flanders a significant background exposure level to airborne mutagenicity, even in areas with limited or no nearby pollution sources. Based on the concentrations of 10 mutagenic PAHs and supposing additivity of their specific mutagenicities, only a few percent (mean 3%) of the observed indirect mutagenic activity could be explained. This implies that most mutagenic activity originated from other substances that were not identified or measured in our chemical analysis. This underscores the importance of bio-monitoring measurements. PMID- 16314140 TI - Assessment of reference values for DNA damage detected by the comet assay in human blood cell DNA. AB - Genotoxicity measured by the comet assay is expressed by different researchers using parameters that are not easy to conceptualize, except for percent tail DNA (%T) or visual score (arbitrary units). A total of 125 publications have reported genotoxicity as DNA damage (representing strand breaks, alkaline labile sites, and transient repair sites), endonuclease III (ENDOIII), or formamidopyrimidine DNA glycosylase (FPG) sensitive sites. I have recalculated the visual score so that it is expressed in the range of 0-100, similar to that of %T. Similar values were obtained for DNA damage and ENDOIII sites, regardless of whether of the data were reported as %T or visual score. Thus, these endpoints can be used interchangeably, assuming that the visual score is expressed in the 0-100 range. Pooled analysis of %T and visual score data showed that the median (25-75%) values of DNA damage, ENDOIII, and FPG sites were 8.6 (4.4-14.5), 11.0 (4.2 19.5), 7.6 (3.2-14.2), respectively. The duration of alkaline treatment and electrophoresis had no significant effect on the level of DNA damage. There was a positive correlation between age and the level of DNA damage. A sub-analysis of DNA damage obtained from European countries showed a negative correlation with latitude. In conclusion, reference values for DNA lesions measured by the comet assay are around 7-11 %T or arbitrary units. PMID- 16314141 TI - A Raman spectroscopic study of selected natural jarosites. AB - Raman spectroscopy has been used to characterise the jarosite group of minerals of formula Mn(Fe3+)6(SO4)4(OH)12 where M may be K, (NH4)+, Na, Ag or Pb and where n = 2 for monovalent cations and 1 for the divalent cations. Raman spectroscopy proved useful for mineral identification especially where closely related minerals crystallise out from solutions where paragenetic relationships exist between the minerals. The band position of the SO4(2-) symmetric stretching mode proved to be a function of the ionic radius of the cation. The bending modes show a slight dependence. The spectra of the natural samples can be complex. This complexity is attributed to the incorporation of low levels of other cations into the structure. PMID- 16314142 TI - Evidence for a wide distribution of negative motor areas in the perirolandic cortex. AB - OBJECTIVE: The perirolandic regions were studied by extensive electrical stimulation to clarify the topography and somatotopic distribution of negative motor areas (NMAs) and examine the clinical significance of these areas. METHODS: We evaluated the cortical function elicited by electrical stimulation in 30 patients with tumors or intractable epilepsy. The somatotopic distribution of NMAs was examined by localizing these regions using Talairach's bicommissural reference system. NMAs within the lesions of two patients were removed under local anesthesia. RESULTS: We obtained negative motor responses following the stimulation of 30 electrodes in 15 patients. On the lateral brain surface, the majority of NMAs for the upper extremities were distributed broadly throughout the premotor cortex, while NMAs for the tongue were only found in the inferior frontal gyrus of the dominant hemisphere. During removal of the NMAs within the lesions of two patients, we documented transient hand clumsiness in one patient. CONCLUSIONS: NMAs were widely distributed throughout the perirolandic area, as well as the previously reported regions in the inferior frontal gyrus. These areas likely function in the control of skilled movements; dysfunction of such movements transiently follows resection of these regions, but is subsequently well compensated for after surgery. SIGNIFICANCE: The localization and consequences of resection of NMAs suggests their clinical significance in motor control. PMID- 16314143 TI - Cortical and thalamic fMRI responses in partial epilepsy with focal and bilateral synchronous spikes. AB - OBJECTIVE: To determine the blood oxygen level-dependent (BOLD) responses to epileptic discharges in the thalamus and cerebral cortex in patients with partial epilepsy. METHODS: Among 64 tested patients, 40 had EEG spikes during scanning and were divided in two groups: unilateral or bilateral independent spikes (29 patients) and bilaterally synchronous spikes (11 patients). Each spike topography was analyzed separately, yielding 40 studies in the first group and 17 in the second. RESULTS: Forty-five percent of focal spike studies showed significant BOLD responses. Cortical activation (positive BOLD) represented the dominant response and had a better correlation with spike location than cortical deactivation (negative BOLD). In the second group, all patients had significant BOLD responses; they were more widespread compared to the first group, and deactivated areas were as important as activated regions. A thalamic response was seen in 12.5% of studies in the first group and 55% in the second. CONCLUSIONS: The thalamus is involved in partial epilepsy during interictal discharges. This involvement and also cortical deactivation are more commonly seen with bilateral spikes than focal discharges. SIGNIFICANCE: These findings show evidence for a role for the thalamus and a more important role for inhibition in secondary bilateral synchrony. PMID- 16314144 TI - Corticospinal disinhibition in paroxysmal kinesigenic dyskinesia. AB - OBJECTIVE: To investigate the underlying mechanism responsible for paroxysmal kinesigenic dyskinesia (PKD). METHODS: We performed a transcranial magnetic stimulation (TMS) study in 12 drug-naive patients with PKD and 10 healthy volunteers. TMS parameters included resting motor threshold, recruitment curve of motor evoked potential amplitudes, short intracortical inhibition and facilitation, long intracortical inhibition (LICI), and silent period. We also measured compound muscle action potential. RESULTS: LICI, representing GABA mediated inhibition, was significantly reduced in the patients (P = 0.033), while results for all other TMS parameters tested were comparable between the two groups. CONCLUSIONS AND SIGNIFICANCE: These results suggest that a specific type of corticospinal inhibitory mechanism is impaired in patients with PKD. PMID- 16314145 TI - Severe dilated cardiomyopathy and quadriceps myopathy due to lamin A/C gene mutation: a phenotypic study. AB - This study reports a family affected by a new phenotype associated with dilated cardiomyopathy and quadriceps myopathy. METHODS: 29 family members underwent a physical and neurological examination, including an electromyogram and biopsy of muscle abnormalities. A cardiac examination was performed in all subjects. RESULTS: The family pedigree (n=72) demonstrated that transmission was autosomal dominant. Eleven subjects had cardiac involvement, only four had quadriceps muscle involvement. Cardiac impairment preceded neurological involvement. The mean age for neurological involvement was 44+/-0.8 years (range 43-45) and cardiac involvement was 37+/-7.9 years (range: 24-45). Cardiac involvement consisted of: hypokinetic dilated cardiomyopathy (64%); atrial fibrillation (100%); ventricular arrhythmias (64%); impaired conduction with bundle branch or complete atrio ventricular block (73%). Four patients required pacemakers and anti arrhythmic therapies. Four patients died: two of refractory heart failure and two of sudden death; two patients were resuscitated following cardiac arrest. Three patients required a prophylactic implantable cardiac defibrillator (ICD). Muscle morphological abnormalities were characterized by a variable number of fibers with rimmed vacuoles. The quadriceps deteriorated progressively without impairment of other muscles. Genotypic study showed a lamin A/C gene mutation. CONCLUSIONS: This family was affected by a new phenotype composed of an autosomal dominant severe dilated cardiomyopathy with conduction defects or arrhythmias and quadriceps myopathy. Cardiac abnormalities preceded neuromuscular disorders and defined the prognosis of this disease. PMID- 16314146 TI - Visual EEG differences in abstinent alcohol dependence and ethnicity. PMID- 16314147 TI - Reversible ventricular arrhythmia in REM sleep associated with hypoxic sleep disordered breathing. PMID- 16314148 TI - Hemodynamic responses to a community-based Tai Chi exercise intervention in ethnic Chinese adults with cardiovascular disease risk factors. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death among older adults worldwide, including Europe, Asia, and North America. In the United States (US), CVD is also the leading cause of death among Asian-Americans. Physical activity has been shown to reduce CVD risk factors. Reduction in blood pressure (BP) in response to Tai Chi (TC) exercise in persons with CVD risk factors have been reported, though not in ethnic Chinese living in the US. AIM: Hemodynamic responses to a 12-week community-based TC exercise intervention among ethnic Chinese with CVD risk factors were examined. METHODS: Quasi-experimental design. Ethnic Chinese > 45 years old with at least 1 major CVD risk factor, living in the San Francisco Bay Area, attended a TC intervention three times a week for 12 weeks. A 2-min step-in-place test assessed aerobic endurance. BP and heart rate were measured at rest, and within 1-min after the step-test. Data were collected at baseline, 6 and 12 weeks. RESULTS: A total of 39 subjects (69% women), 66 +/- 8.3 years old, with hypertension (92%), hypercholesteremia (49%), and/or diabetes (21%), and 1 current smoker participated. Adherence to the intervention was high (87%). Subjects were sedentary at baseline, though had a statistically significant improvement in aerobic endurance over-time (eta2 = 0.39). At baseline, the average BP at rest was 150/86, while BP in response to the step-test was 178/99. Clinically and statistically significant reductions in BP at rest (131/77), and in response to the step-test (164/82) were found over 12 weeks of TC (p < 0.01). No significant change in heart rate was observed. CONCLUSIONS: This innovative, culturally relevant, community-based 12-week TC exercise intervention, appealed to Chinese adults with CVD risk factors, with significant reductions in BP and improvement in aerobic endurance. Given the number of persons estimated to have HTN and other CVD risk factors, the identification of new approaches to improve health, combined with risk factor reduction is needed. This is particularly important, given the rise in HTN among adults in the US and the associated public health burden of HTN. TC has the potential to reduce expenditures associated with CVD by facilitating a lifestyle that promotes physical activity, while remaining a low-tech, low-cost alternative to exercise. PMID- 16314149 TI - The antiepileptic drug phenytoin affects sodium transport in toad epithelium. AB - The effects of phenytoin on isolated Pleurodema thaul toad skin were investigated. Low (micromolar) concentrations of the antiepileptic agent applied to the outside surface of the toad epithelium increased the electrical parameters (short-circuit current and potential difference) by over 40%, reflecting stimulation of Na(+) transport, whereas higher (millimolar concentrations, outside and inside surface) decreased both electric parameters, the effect being greater at the inside surface (40% and 80% decrease, respectively). The amiloride test showed that the stimulatory effect was accompanied by an increase and the inhibitory effect by a decrease in the sodium electromotive force (ENa). It is concluded that the drug interaction with membrane lipid bilayers might result in a distortion of the lipid-protein interface contributing to disturbance of Na(+) epithelial channel activity. After applying the Na(+)-K(+)-ATPase blocker ouabain and replacing the Na(+) ions in the outer Ringer's solution by choline, it was concluded that both active and passive transport are involved in sodium absorption, although active transport predominates. PMID- 16314150 TI - Applicability of the Rockall score in patients undergoing endoscopic therapy for upper gastrointestinal bleeding. AB - BACKGROUND: The Rockall score is used to assess the prognosis of patients with upper gastrointestinal bleeding. AIM: To assess the applicability of the Rockall score in patients undergoing endoscopic therapy for upper gastrointestinal bleeding. METHODS: Retrospective evaluation of the Rockall score in the period 1995-2001. To evaluate the applicability of the Rockall system, two groups were created: group I (Rockallor=6 points). RESULTS: Two hundred and twenty-two patients were included. The median age of patients was 65 +/ -17 years. Hypotension and associated diseases were present in 20 and 50% of patients, respectively. Re-bleeding occurred in 50 patients (23%) whose median score was 7, whereas the median score of patients without re bleeding was 6 (p=0.14). There were 20 deaths (9%) with a median score of 8, whilst the median score of surviving patients was 6 (p<0.001). Sixteen patients in group I (18.4%) and 34 in group II (25.2%) re-bled (p=0.25). All the patients who died belong to group II with a Rockall score>or=6 (15% versus 0% in groups II and I, respectively, p<0.001). CONCLUSION: The Rockall score can be used in patients who undergo therapeutic endoscopy for upper gastrointestinal bleeding to identify those with high risk for mortality. PMID- 16314151 TI - Multiple gastric tube carcinomas after curative oesophagectomy. PMID- 16314152 TI - Endoscopic ultrasound and computer tomography are inaccurate methods of classifying cystic pancreatic lesions. AB - BACKGROUND: Despite advances in imaging modalities, preoperative diagnosis of pancreatic cystic lesions remains difficult. AIM: To assess the accuracy of endoscopic ultrasound and computer tomography to preoperatively distinguish benign from potentially malignant and malignant pancreatic cystic lesions. METHODS: Photograph series obtained from endoscopic ultrasound examinations of 66 patients with cystic pancreatic lesions were blindly reviewed by three endoscopic ultrasonographers. Forty-one of those 66 patients also underwent a computer tomography scan at our institution, which was blindly reviewed by a single radiologist. Computer tomography and endoscopic ultrasound classification into benign and malignant and potentially malignant pancreatic cystic lesions was correlated with the final diagnosis, which was established by surgical pathology (n = 43), diagnostic fine needle aspiration (n = 13) or follow-up imaging (n = 10). Interobserver agreement was measured using kappa statistics. RESULTS: Endoscopic ultrasound classification by the three examiners into benign versus malignant or potentially malignant cystic lesions was correct in 65-67%. Interobserver agreement was 50%. Kappa values for pairs of endoscopic ultrasound examiners were 0.16, 0.43 and 0.53. Computer tomography classification was correct in 71% and in agreement with the endoscopic ultrasound classification in 56-61% (kappa 0.12 to 0.27). CONCLUSIONS: Endoscopic ultrasound and computer tomography cannot accurately distinguish between benign pancreatic cystic lesions and malignant or potentially malignant ones. There is poor-to-modest interobserver agreement in classifying these lesions. PMID- 16314153 TI - Apolipoprotein C-II is a novel substrate for matrix metalloproteinases. AB - We previously reported an efficient proteomic approach to identify matrix metalloproteinase (MMP) substrates from complex protein mixture. Using the proteomic approach, apolipoprotein C-II (apoC-II), which is a cofactor of lipoprotein lipase (LPL) and a component of very-low density lipoprotein and chylomicron, has been identified as a putative MMP-14 substrate. Cleavage of apoC II, with various MMPs, demonstrated that apoC-II is cleaved most efficiently by MMP-14, and also by MMP-7, among the tested MMPs. The 79-amino acid residue apoC II was cleaved between Asn35 and Leu36 by MMP-14, and between Phe14 and Leu15 and between Asn35 and Leu36 by MMP-7. Cleavage of apoC-II by MMP-14 markedly decreased LPL activity and would thus impair hydrolysis of triglycerides in plasma and transfer of fatty acids to tissues. Our result suggests that cleavage of apoC-II by MMPs would be important for development of pathophysiological situations of apoC-II deficiency such as atherosclerosis. PMID- 16314154 TI - The prolonged half-lives of new erythropoietin derivatives via peptide addition. AB - Erythropoietin, or Epo, is a hematopoietic cytokine that promotes erythropoiesis, and recombinant human Epo has been used in the treatment of anemia in various chronic diseases. Here, we have constructed novel Epo derivatives with prolonged half-lives by adding peptides to the carboxy terminus of Epo without using linkers. The fused peptides were selected from the carboxy terminal region of human chorionic gonadotropin (hCG) or human thrombopoietin (hTpo), which promote the proper folding, secretion, and stabilization of bioactive glycoproteins. Addition of these peptides did not interfere with secretion or receptor binding, and significantly increased the in vivo half-life of human Epo, as measured by intravenous administration in rats. The plasma half-life of the Epo constructs was longest when the carboxy terminal 28 aa of the beta subunit of hCG was added (Epo-CGC), a half-life that was slightly longer than NESP (Aranesp), which is the most effective Epo product in current clinical use. The transformation of four Ser glycosylation sites to Ala on the CGC sequence also lengthened the plasma half-life of Epo, indicating that the in vivo stabilizing effect of the hCG peptide was due to both structures within the peptide itself and its O glycosylations. The application of the carboxy terminal half of hTpo also resulted in remarkably reduced elimination of the Epo chimera (Epo-TpC), possibly due to protection by the TpC sequence. The in vivo hematopoietic activity of Epo derivatives in mice was consistent with their pharmacokinetic profiles. Therefore, these derivatives with prolonged half-lives may provide opportunities for developing new Epo therapeutics with less frequent administration. PMID- 16314155 TI - Simultaneous picogram determination of "earthy-musty" odorous compounds in water using solid-phase microextraction and gas chromatography-mass spectrometry coupled with initial cool programmable temperature vaporizer inlet. AB - "Earthy-musty" off-flavor problem in water samples are due to organic compounds present at the sub-part-per-trillion level. Most of the developments in the analysis of tastes and odorous compounds focus on the extraction pre concentration technique, with detection at picogram per liter level of the earthy musty off-flavor compounds difficult to be achieved. In this study, a simple, efficient and sensitive method for the analysis of odorous compounds has been developed by the application of solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) with initial cool programmable temperature vaporizer (PTV) inlet for the first time. Compared with initial hot PTV inlet, the initial cool inlet could greatly improve the system sensitivity, especially for the compounds with good volatility, e.g. 2-methylisoborneol (MIB). StableFlex divinylbenzene/Carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber was found to possess the best extraction efficiency towards these odorous compounds in water. Various SPME and PTV conditions have been studied and optimized in detail. The optimized method has been validated with good linearity, precision and accuracy. The method detection limits (MDL) of the targeted odorous compounds were found to be 0.32ng/L for 2,4,6-trichloroanisole (2,4,6-TCA), 0.14ng/L for 2,3,6-trichloroanisole (2,3,6-TCA), 0.16ng/L for 2,3,4-trichloroanisole (2,3,4 TCA), 0.38ng/L for 2,4,6-tribromoanisole (2,4,6-TBA), 0.16ng/L for gesomin and 0.15ng/L for MIB. To the best of our knowledge, this represents the best sensitivity achieved for analysis of gesomin and MIB in water via the simple and efficient SPME method. The current method has been successfully applied in the analyses of different water samples. PMID- 16314156 TI - Hydrodistillation-headspace solvent microextraction, a new method for analysis of the essential oil components of Lavandula angustifolia Mill. AB - A new method involving concurrent headspace solvent microextraction combined with continuous hydrodistillation (HD-HSME) for the extraction and pre-concentration of the essential oil of Lavandula angustifolia Mill. into a microdrop is developed. A microdrop of n-hexadecane containing n-heptadecane (as internal standard) extruded from the needle tip of a gas chromatographic syringe was inserted into the headspace above the plant sample. After extraction for an optimized time, the microdrop was retracted into the syringe and injected directly into a GC injection port. The effects of the type of extracting solvent, sample mass, microdrop volume and extraction time on HD-HSME efficiency were investigated and optimized. Using this method, thirty-six compounds were extracted and identified. Linalool (32.8%), linalyl acetate (17.6%), lavandulyl acetate (15.9%), alpha-terpineol (6.7%) and geranyl acetate (5.0%) were found to be the major constituents. To the best of our knowledge this is the first report on the use of continuous headspace solvent microextraction coupled with hydrodistillation for investigation of essential oil components. PMID- 16314157 TI - Development of a solid phase extraction for 13 'new' generation antidepressants and their active metabolites for gas chromatographic-mass spectrometric analysis. AB - A solid phase extraction procedure (SPE) for 13 'new' antidepressants (venlafaxine, fluoxetine, viloxazine, fluvoxamine, mianserin, mirtazapine, melitracen, reboxetine, citalopram, maprotiline, sertraline, paroxetine and trazodone) together with eight of their metabolites (O-desmethylvenlafaxine, norfluoxetine, desmethylmianserine, desmethylmirtazapine, desmethylcitalopram, didesmethylcitalopram, desmethylsertraline and m-chlorophenylpiperazine) from plasma is optimized using HPLC-DAD as monitoring system. Special attention has been paid to the choice of washing and eluting solvent, resulting in a highly concentrated, clean and moisture free extract, also suitable for GC-MS. A total number of 10 sorbents (apolar, polymeric, ion-exchange and mixed mode) was evaluated. Based on recovery, reproducibility and absence of interfering substances the strong cation exchanger gave the best results. Recoveries were determined at low and high therapeutic and toxic levels and ranged between 70 and 109% for all compounds, except for trazodone (39%). PMID- 16314158 TI - Determination of phenols in water samples by single-drop microextraction followed by in-syringe derivatization and gas chromatography-mass spectrometric detection. AB - Trace analysis of phenolic compounds in water was performed by coupling single drop microextraction (SDME) with in-syringe derivatization of the analytes and GC MS analysis. The analytes were extracted from a 3ml sample solution using 2.5microl of hexyl acetate. After extraction, derivatization was carried out in syringe barrel using 0.5microl of N,O-bis(trimethylsilyl)acetamide. The influence of derivatizing reagent volume, derivatization time and temperature on the yield of the in-syringe silylation was investigated. Derivatization reaction is completed in 5min at 50 degrees C. Experimental SDME parameters, such as selection of organic solvent, sample pH, addition of salt, extraction time and temperature of extraction were studied. Analytical parameters, such as enrichment factor, precision, linearity and detection limits were also determined. The limits of detection were in the range of 4-61ng/l (S/N=3). The relative standard deviations obtained were between 4.8 and 12% (n=5). PMID- 16314159 TI - Analysis of pesticides in fruits by pressurized liquid extraction and liquid chromatography-ion trap-triple stage mass spectrometry. AB - A multi-residue method using pressurized liquid extraction (PLE) and liquid chromatography-quadrupole ion trap-triple stage mass spectrometry (LC-IT-MS(3)) has been developed for determining trace levels of pesticides in fruits. The selected pesticides can be distinguished in: benzimidazoles and azoles, organophosphorus, carbamates, neonicotinoids, and acaricides. PLE has been optimized to extract these pesticide residues from oranges and peaches by studying the effect of experimental variables on PLE efficiency. Samples were extracted at high temperature and pressure (75 degrees C and 1500psi) using ethyl acetate as extraction solvent and acidic alumina as drying agent. The recoveries obtained by PLE ranged from 58% to 97% and the relative standard deviation (RSDs) from 5% to 19%. The limits of quantification (LOQs) of the compounds were from 0.025 to 0.25mgkg(-1), which are well-below the maximum residue limits (MRLs) established by the European Union (EU) and the Spanish legislations. PMID- 16314160 TI - Prediction of retention times of proteins in hydrophobic interaction chromatography using only their amino acid composition. AB - This paper focuses on the prediction of the dimensionless retention time of proteins (DRT) in hydrophobic interaction chromatography (HIC) by means of mathematical models based, essentially, only on aminoacidic composition. The results show that such prediction is indeed possible. Our main contribution was the design of models that predict the DRT using the minimal information concerning a protein: its aminoacidic composition. The performance is similar to that observed in models that use much more sophisticated information such as the three-dimensional structure of proteins. Three models that, in addition to the amino acid composition, use different assumptions about the amino acids tendency to be exposed to the solvent, were evaluated in 12 proteins with known experimental DRT. In all the cases analyzed, the model that obtained the best results was the one based on a linear estimation of the aminoacidic surface composition. The models were adjusted using a collection of 74 vectors of aminoacidic properties plus a set of 6388 vectors derived from these using two mathematical tools: k-means and self-organizing maps (SOM) algorithms. The best vector was generated by the SOM algorithm and was interpreted as a hydrophobicity scale based partly on the tendency of the amino acids to be hidden in proteins. The prediction error (MSE(JK)) obtained by this model was almost 35% smaller than that obtained by the model that supposes that all the amino acids are completely exposed and 40% smaller than that obtained by the model that uses a simple correction factor considering the general tendency of each amino acid to be exposed to the solvent. In fact, the performance of the best model based on the aminoacidic composition was 5% better than that observed in the model based on the three-dimensional structure of proteins. PMID- 16314161 TI - Influence of mobile phase composition and thermodynamics on the normal phase chromatography of echinocandins. AB - In the normal phase preparative HPLC of fermentation derived echinocandins, resolution of key impurities from the product of interest, pneumocandin B(o), is accomplished using a ternary ethyl acetate/methanol/water mobile phase with silica gel as the sorbent. In this work, previous characterization of the system is extended to define the impact and role of water content on the separation efficiency and retention of pneumocandin B(o). Experimental results indicate that column efficiency, measured using both the product of interest and small molecule tracers (compounds used for pulse tests), is good despite the use of an irregular silica and unusually high levels (greater than 6%) of water in the mobile phase. In contrast to column efficiency measurements using small molecules (MEK and toluene), measurements performed with the product itself indicate improved efficiency with increasing water content of the mobile phase. Building on these results, a scale-up/scale-down protocol was developed based on measurements of column efficiency using theoretical plate counts determined with pneumocandin B(o). Since the solubility of pneumocandin B(o) in the ternary mobile phase is relatively low, a higher strength solvent with higher levels of methanol and water is employed for dissolution of the crude product at concentrations of up to 40g/L. The mismatch between the high strength solvent used for the feed introduction and the mobile phase has the potential to affect column performance. The impact of this mismatch using plate count measurements with the product at both analytical and semi-preparative scales was found not to be significant. Finally, a van't Hoff analysis was performed to characterize the thermodynamics of adsorption of pneumocandin B(o) on silica. The analysis shows that the adsorption process for pneumocandin B(o) on silica in the ternary solvent system is endothermic (DeltaH(ads)>0), implying that the adsorption is entropically driven. Results from an overall water balance across the column indicate significant enrichment of adsorbed water on the silica surface. These results further emphasize the importance of selective partitioning of water between the bulk mobile phase and the silica as a dominant factor in controlling retention. PMID- 16314162 TI - Separation and purification of chlorogenic acid by molecularly imprinted polymer monolithic stationary phase. AB - Separation and purification of chlorogenic acid by removal of the impurities compounds co-existed in the product using molecular imprinting technique was firstly reported. In this work, an in situ synthesis method was utilized for the preparation of molecularly imprinted polymer monolithic stationary phase using the impurity molecule (caffeic acid) as template, the mixture of tetrahydrofuran and isooctane as solvent, and methacrylic acid and ethyl glycol dimethacrylate as functional monomer and cross-linker, respectively. The retention behavior of the monolithic polymer to chlorogenic acid molecule, the template and several main impurities compounds in the product was studied and the adsorption capacity of compounds on the stationary phase determined by frontal chromatographic technique. A relatively weak retention of the target product molecule (chlorogenic acid) on the polymer and a strong adsorption capability of the monolith to the template and several main impurities were observed. This might mainly result from the 'shape' difference of chlorogenic acid molecule with the impurities compounds molecules. This approach was shown to be successful for the separation and purification of chlorogenic acid from the extract of Eucommia ulmodies leaves. PMID- 16314163 TI - High-performance liquid chromatographic enantioseparation of bicalutamide and its related compounds. AB - Direct high-performance liquid chromatographic methods were developed for the enantioseparation of (R,S)-bicalutamide (1) and its analogs (+/-)-3-chloro-N-(4 cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide (2), (+/-)-N-(4 cyano-3-(trifluoro-methyl)phenyl)-2-methyloxirane-2-carboxamide (3), (+/-)-4 fluorophenylsulfonyl-2-hydroxy-2-methylpropionic acid (4) and (+/-)-3-hydroxy-N (4-cyano-3-(trifluoromethyl)phenyl)-2-hydroxy-2-methylpropanamide (5). The methods involved the use of a cellulose-based Chiralcel OD-H, macrocyclic glycopeptide-based Chirobiotic T, TAG and R, beta-cyclodextrin-based Cyclobond I 2000SN and t-butyl carbamate-derivatized quinine-based columns. The conditions affording the best resolution were found by selection and variation of the mobile phase compositions, and the differences in separation capability of the methods were noted. The sequence of elution of the enantiomers was determined in all cases. PMID- 16314164 TI - Effect of the endcapping of reversed-phase high-performance liquid chromatography adsorbents on the adsorption isotherm. AB - The retention mechanisms of n-propylbenzoate, 4-t ert-butylphenol, and caffeine on the endcapped Symmetry-C(18) and the non-endcapped Resolve-C(18) are compared. The adsorption isotherms were measured by frontal analysis (FA), using as the mobile phase mixtures of methanol or acetonitrile and water of various compositions. The isotherm data were modeled and the adsorption energy distributions calculated. The surface heterogeneity increases faster with decreasing methanol concentration on the non-endcapped than on the endcapped adsorbent. For instance, for methanol concentrations exceeding 30% (v/v), the adsorption of caffeine is accounted for by assuming three and two different types of adsorption sites on Resolve-C(18) and Symmetry-C(18), respectively. This is explained by the effect of the mobile phase composition on the structure of the C(18)-bonded layer. The bare surface of bonded silica appears more accessible to solute molecules at high water contents in the mobile phase. On the other hand, replacing methanol by a stronger organic modifier like acetonitrile dampens the differences between non-endcapped and endcapped stationary phase and decreases the degree of surface heterogeneity of the adsorbent. For instance, at acetonitrile concentrations exceeding 20%, the surface appears nearly homogeneous for the adsorption of caffeine. PMID- 16314165 TI - Comprehensive two-dimensional liquid chromatography with on-line Fourier transform-infrared-spectroscopy detection for the characterization of copolymers. AB - The on-line coupling of comprehensive two-dimensional liquid chromatography (liquid chromatography x size-exclusion chromatography, LC x SEC) and infrared (IR) spectroscopy has been realized by means of an IR flow cell. The system has been assessed by the functional-group analysis of a series of styrene methylacrylate (SMA) copolymers with varying styrene content. Ultraviolet (UV) detection was used as a detection technique to verify the detection with IR. The LC x SEC-IR functional-group contour plots (comprehensive chromatograms) obtained for styrene were in agreement with the contour plots constructed from the UV signal. In addition, contour plots can be obtained from non-UV-active groups. One such plot, for the carbonyl-stretching vibration of methylacrylate (MA), is shown. Selective detection of MA proved possible using flow cell IR detection. The combination of the contour plots for styrene and MA allowed a full characterization of the copolymer and it was revealed that the present series of SMA copolymers exhibited homogeneous chemical-composition distributions (CCDs). In addition, commercially available fast-SEC columns have been assessed in this study with respect to their potential to serve as second-dimension separation columns. PMID- 16314166 TI - Determination of biogenic amines in chocolate by ion chromatographic separation and pulsed integrated amperometric detection with implemented wave-form at Au disposable electrode. AB - A rapid and selective cation exchange chromatographic method coupled to integrated pulsed amperometric detection (PAD) has been developed to quantify biogenic amines in chocolate. The method is based on gradient elution of aqueous methanesulfonic acid with post column addition of strong base to obtain suitable conditions for amperometric detection. A potential waveform able to keep long time performance of the Au disposable electrode was set up. Total analysis time is less than 20min. Concentration levels of dopamine, serotonin, tyramine, histamine and 2-phenylethylamine were measured, after extraction with perchloric acid from 2g samples previously defatted twice with petroleum ether. The method was used to determine the analytes in chocolate real matrices and their quantification was made with standard addition method. Only dopamine, histamine and serotonin were found in the analysed real samples. Repeatabilities of their signals, computed on their amounts in the real samples, were 5% for all of them. Repeatabilities of tyramine and phenethylamine were relative to standard additions to real samples (close to 1mg/l in the extract) and were 7 and 3%, respectively. Detection limits were computed with the 3s of the baseline noise combined with the calibration plot regression parameters. They were satisfactorily low for all amines: 3mg/kg for dopamine, 2mg/kg for tyramine, 1mg/kg for histamine, 2mg/kg for serotonin, 3mg/kg for 2-phenylethylamine. PMID- 16314167 TI - Selective high performance liquid chromatography imprinted-stationary phases for the screening of phenylurea herbicides in vegetable samples. AB - Molecularly imprinted polymers (MIPs) for the determination of phenylurea herbicides have been synthesized by polymerisation of the appropriated reagents mixture within the pores of preformed spherical silica particles leading to a silica-MIP composite material. Subsequently, the silica matrix was etched away resulting in MIP beads which can be considered the "mirror image" of the original silica mold. The MIP particles were packed in stainless steal HPLC columns (125mmx4.6mm I.D.) and the materials were evaluated as imprinted-stationary phases for phenylurea herbicides. The imprinting effect of the originated specific binding sites for the selective recognition of phenylurea herbicides was clearly demonstrated. An efficient separation of a mixture of phenylurea herbicides in two groups, with or without a methoxy group in the chemical structure, was achieved and well shaped and defined peaks were obtained. Finally, the optimum imprinted column (prepared using linuron as template, 2 (trifluoromethyl)-acrylic acid as monomer, 72h of polymerisation time and the subsequent dissolution of silica matrix) was used for the LC-UV screening of phenylurea herbicides directly from vegetable sample extracts without any previous clean-up step at low concentration level in less than 10min. PMID- 16314168 TI - Relevance of pi-pi and dipole-dipole interactions for retention on cyano and phenyl columns in reversed-phase liquid chromatography. AB - Previous work suggests that pi-pi interactions between certain solutes and both phenyl and cyano columns can contribute to sample retention and the selectivity of these two column types versus alkylsilica columns. Recent studies also suggest that dipole-dipole interactions are generally unimportant for retention on cyano columns. The present study presents data for 44 solutes, three columns and two different mobile phases that were selected to further test these conclusions. We find that pi-pi interactions can contribute to retention on both cyano and phenyl columns, while dipole-dipole interactions are likely to be significant for the retention of polar aliphatic solutes on cyano columns. When acetonitrile/water mobile phases are used, both pi-pi and dipole-dipole interactions are suppressed, compared to the use of methanol/water. PMID- 16314169 TI - Oscillatory transverse electric field enhances mass transfer and protein capacity in ion-exchange electrochromatography. AB - Ion-exchange electrochromatography with an oscillatory electric field perpendicular to mobile-phase flow driven by pressure (pIEEC) was developed with a column design of rectangle cross-section. The effect of electric field strength on the dynamic binding capacity (DBC) was examined by frontal analysis of bovine serum albumin (BSA) adsorption to the packed beds of DEAE Sepharose FF in Tris glycine buffer (pH 8.2). It was shown that the DBC at 10% breakthrough (Q(10)) in the pIEEC increased linearly with increasing the electric field strength. For example, with a packed-bed height of 15mm and electric potential gradient of 38V/cm, Q(10) increased four times over that in normal ion-exchange chromatography. So, the transverse electric field has created significant electro kinetic mass transports (electroosmosis and electrophoresis) that intensified exterior liquid-film and intraparticle mass transfers, leading to the increased protein binding capacity. Due to the increased capacity in the pIEEC, partial resolution of BSA and IgG under an overload condition was realized without any process optimization. The results have revealed that an electric potential gradient of 20V/cm was enough to greatly enhance the DBC in the pIEEC, and when necessary, high electric field strength can be realized with a low applied voltage because the side distance of the column is usually an order of magnitude smaller than its height. The use of low voltage to carry out electrochromatography is a significant advantage of the pIEEC over conventional electrochromatography with axial electric field. PMID- 16314170 TI - Direct quantitative determination of cyanamide by stable isotope dilution gas chromatography-mass spectrometry. AB - Cyanamide is a multifunctional agrochemical used, for example, as a pesticide, herbicide, and fertilizer. Recent research has revealed that cyanamide is a natural product biosynthesized in a leguminous plant, hairy vetch (Vicia villosa). In the present study, gas chromatography-mass spectrometry (GC-MS) equipped with a capillary column for amines was used for direct quantitative determination of cyanamide. Quantitative signals for ((14)N(2))cyanamide, ((15)N(2))cyanamide (internal standard for stable isotope dilution method), and m (trifluoromethyl)benzonitrile (internal standard for correcting errors in GC-MS analysis) were recorded as peak areas on mass chromatograms at m/z 42 (A(42)), 44 (A(44)), and 171 (A(IS)), respectively. Total cyanamide content, ((14)N(2))cyanamide plus ((15)N(2))cyanamide, was determined as a function of (A(42)+A(44))/A(IS). Contents of ((14)N(2))cyanamide and ((15)N(2))cyanamide were then calculated by multiplying the total cyanamide content by A(42)/(A(42)+A(44)) and A(44)/(A(42)+A(44)), respectively. The limit of detection for the total cyanamide content by the GC-MS analysis was around 1ng. The molar ratio of ((14)N(2))cyanamide to ((15)N(2))cyanamide in the injected sample was equal to the observed A(42)/A(44) value in the range from 0.1 to 5. It was, therefore, possible to use the stable isotope dilution method to quantify the natural cyanamide content in samples; i.e., the natural cyanamide content was derived by subtracting the A(42)/A(44) ratio of the internal standard from the A(42)/A(44) ratio of sample spiked with internal standard, and then multiplying the resulting difference by the amount of added ((15)N(2))cyanamide (SID-GC-MS method). This method successfully gave a reasonable value for the natural cyanamide content in hairy vetch, concurring with the value obtained by a conventional method in which cyanamide was derivatized to a photometrically active compound 4-cyanimido-1,2 naphthoquinone and analyzed with reversed-phase HPLC (CNQ-HPLC method). The determination range of cyanamide in the SID-GC-MS method was almost the same as that in the CNQ-HPLC method; however, the SID-GC-MS method was much simpler than the CNQ-HPLC method. PMID- 16314171 TI - Identification of fatty acids in fishes collected from the Ohio River using gas chromatography-mass spectrometry in chemical ionization and electron impact modes. AB - Analyses of fatty acid (FA) composition in freshwater fishes promote understanding of the potential relationship between fish health or human nutrition and specific FAs. Therefore, the chemical identity of FAs in endemic fishes must be established. Paddlefish, sauger, and white bass were collected from the Ohio River. The structural identification of esterified FAs from fish fillet lipids was conducted using gas chromatography-mass spectrometry (GC-MS). The same 13 FAs, composing more than 90% of the mass of FAs extracted by techniques used in this research, were found in all three species examined. Carbon chain length and degree and position of unsaturation were determined from the characteristic ionization and fragmentation of FA methyl esters (FAMEs) resulting from GC-MS electron impact (EI) and chemical ionization (CI) modes. Assignment of structure to the extracted FAs required complementary interpretation of both EI and CI MS. The EI spectra observed substantiate findings reported in the literature. The novelty of this research is in the thorough interpretation of CI spectra for which less data are available. The observations reported for analyses of fishes will be useful to all researchers studying FAs regardless of sample media. PMID- 16314172 TI - Determination of the limits of identification and quantitation of selected organochlorine and organophosphorous pesticide residues in surface water by full scan gas chromatography/mass spectrometry. AB - In this work, we report a reliable method for quantitation and determination of the limits of identification of 14 organochlorine and organophosphorous pesticide (OPP) residues in surface water. The method features the simultaneous identification and quantitation of targeted pesticides and the possibility of identification of any other eluting compounds. The method is based on liquid liquid extraction (LLE) with a mixture of petroleum ether and dichloromethane (70:30, v/v) followed by gas chromatographic separation and a full-scan mass spectrometric detection (GC-MS). The method presents a new validation parameter, limit of identification (LOI) which is defined for our purpose as the lowest analyte concentration that yields a library searchable mass spectrum. The method is linear over the range 0.048-1.20microgL(-1) for nine pesticides and 0.024 0.60microgL(-1) for the other five pesticides. Correlation coefficients vary between 0.988 and 0.998. Limits of detection (LODs) vary between 0.005 and 0.05microgL(-1) for 4,4'-DDT and LOIs vary between 0.012 and 0.048microgL(-1). PMID- 16314173 TI - Analysis of chemical warfare agents III. Use of bis-nucleophiles in the trace level determination of phosgene and perfluoroisobutylene. AB - The reactivity of phosgene and perfluoroisobutylene (PFIB) towards 1,2-bis nucleophiles was exploited to allow determination of these gases in air samples. 2-Aminothiophenol (ATP), 3,4-dimercaptotoluene (DMT) and 2 hydroxymethylpiperidine (HMP) were evaluated as bis-nucleophiles capable of forming thermally-stable derivatives with phosgene and PFIB when loaded with triethylamine onto Tenax TA. Experimental design was used to optimise thermal desorption conditions. Detection limits in the low ngm(-3) range were observed for the five derivatives investigated. This work represents the most sensitive analytical method for trace level quantitation of phosgene and PFIB published to date. PMID- 16314174 TI - Separation and determination of norepinephrine, epinephrine and isoprinaline enantiomers by capillary electrophoresis in pharmaceutical formulation and human serum. AB - A capillary electrophoresis method with ultraviolet (UV) detection was developed and optimized for the enantiomer separation of norepinephrine (NE), epinephrine (EP) and isoprenaline (IP) using dual cyclodextrins (CDs) of 2-hydroxypropyl-beta CD (HP-beta-CD) and heptakis (2,6-di-o-methyl)-beta-CD (DM-beta-CD) as chiral selectors. Optimal separation was obtained using a running buffer of 50mM phosphate containing 30mM HP-beta-CD and 5mM DM-beta-CD at pH 2.90 and a field strength of 20kV in 45cmx75mum (40cm effective length) uncoated capillary. The UV absorbance detection was set at 205nm. A 0.1% (w/w) polyethylene glycol or 0.1% (v/v) acetonitrile was used to enhance the detection sensitivity. There was a wide and excellent linear calibration graph for each enantiomer in the range 1.0x10(-3) to 1.0x10(-6)M and the detection limit (S/N=3) was found from 8.5x10( 7) to 9.5x10(-7)M. The method has been applied for the determination of isoprenaline in isoprenaline hydrochloride aerosol and to the analysis of serum samples. The recoveries of NE and EP in serum and IP in drug were ranged from 90 to 110%. The relative standard deviations of all the analyte peaks were less than 2.8% for migration time and less than 4.8% for peak area. PMID- 16314175 TI - Separation of caffeine and theophylline in poly(dimethylsiloxane) microchannel electrophoresis with electrochemical detection. AB - A method for the rapid separation and sensitive determination of caffeine and theophylline was presented in poly(dimethylsiloxane) (PDMS) microchannel electrophoresis integrated with electrochemical detection. By using methanol as an additive, the peak shape and resolution were essentially improved. The analytes were well separated within only 40s in the running buffer of 5.0mM borate solution (pH 9.2) containing 10% (v/v) methanol. The linear ranges were from 6microM to 0.6mM and the detection limits were 4microM for caffeine and theophylline, respectively. The proposed method has been successfully applied to determine caffeine and theophylline in rat serum and urine. PMID- 16314176 TI - Separation and determination of tetrandrine and fangchinoline in herbal medicines by flow injection-micellar electrokinetic capillary chromatography with internal standard method. AB - A simple, rapid and precision flow injection-micellar electrokinetic capillary chromatography (FI-MEKC) system with trimethoprim as internal standard (IS) for automated quantitative analysis of tetrandrine (TET) and fangchinoline (FAN) in various herbal medicines was demonstrated. The real sample throughput was 19-40 samples per hour using the background electrolyte (BGE) containing 15mM acetic acid-15mM sodium acetate-3% (v/v) polyoxyethylene sorbitan monolaurate (Tween 20) 5% (v/v) methanol at pH 5.5. The method resulted in excellent linearity, with correlation coefficient of regression equation of 0.9996 and 0.9991 for TET and FAN, respectively. Recoveries were in the range 95-109% and 92-106% for TET and FAN, respectively. PMID- 16314178 TI - Stable incorporation of sequence specific repressors Ash1 and Ume6 into the Rpd3L complex. AB - Histone deacetylation by Saccharomyces cerevisiae Rpd3 represses genes regulated by the Ash1 and Ume6 DNA-binding proteins. Rpd3 exists in a small 0.6 MDa (Rpd3S) and large 1.2 MDa (Rpd3L) corepressor complex. In this report, we identify by mass spectrometry and MudPIT the subunits of the Rpd3L complex. These included Rpd3, Sds3, Pho23, Dep1, Rxt2, Sin3, Ash1, Ume1, Sap30, Cti6, Rxt3 and Ume6. Dep1 and Sds3, unique components of Rpd3L, were required for Rpd3L integrity and HDAC activity. Similar to RPD3, deletion of DEP1 enhanced telomeric silencing and derepressed INO1. Two sequence-specific repressors, Ash1 and Ume6, were stably associated with Rpd3L. While both of these proteins localized to the INO1 and HO promoters, the repression of these genes were dependent only on Ume6 and Ash1, respectively. Thus, the Rpd3L complex is directly recruited to specific promoters through multiple integral DNA-binding proteins. PMID- 16314179 TI - Rotavirus vaccine: early introduction in Latin America-risks and benefits. AB - Rotavirus infection is the cause of severe gastroenteritis of young children worldwide, leading to an estimate of 600,000 deaths a year. Efforts to develop an effective and safe vaccine resulted in licensing in 1998 of a live oral vaccine (RotaShield) that was withdrawn less than 1 year later when reports of cases of intussusception were linked to its application. This led to development of new rotavirus vaccine candidates that are currently in late phase III clinical trials. One of these vaccines, GlaxoSmithKline's Rotarix, was licensed in July 2004 to be used in Mexico. This review describes the general background for rotavirus vaccine development, the different vaccine candidates that have been tested or are currently being evaluated, the association of rotavirus vaccination with the bowel blockage known as intussuception, and discusses the benefits and risks of the fast-track introduction of Rotarix in Latin America, and particularly in Mexico. PMID- 16314180 TI - Role of glutamate transporters in the clearance and release of glutamate during ischemia and its relation to neuronal death. AB - Glutamate neurotransmitter action on postsynaptic receptors is terminated by its clearance from the synaptic cleft by transporter proteins located in neurons and glial cells. Failure of glutamate removal can lead to neuronal death due to its well-known neurotoxic properties. Glutamate transporters are dependent on external Na+, and thus on the activity of Na+/K+ ATPases, which maintain the Na+ concentration gradient. When the energy brain requirements are not fulfilled by the appropriate blood supply of glucose and oxygen, the Na+ gradient collapses leading to impaired glutamate and aspartate removal, or even to the release of these amino acids through the reverse operation of their transporters. Such a scenario would be associated with brain ischemia and hypoglycemia due to the prompt decline in ATP levels. In addition, some evidence suggests that downregulation of glutamate transporters after the ischemic period, or the dysfunction induced by oxidation, contributes to the accumulation of extracellular glutamate and neuronal death. Neuronal damage is associated with excitotoxicity, a type of cell death triggered by the overactivation of glutamate receptors and the loss of calcium homeostasis. Throughout this review we will discuss recent evidence suggesting that failure of glutamate transport during ischemia contributes to the elevation of extracellular glutamate and to the induction of excitotoxicity. We will also discuss the contribution of glial vs. neuronal glutamate transporters in ischemic damage, and the involvement of the different glutamate transporter subtypes. We will focus on experimental data from rodent models, because many of the studies on glutamate transport and ischemic damage have been performed in these animal species. PMID- 16314181 TI - Protective effect of nitric oxide pathway in resveratrol renal ischemia reperfusion injury in rats. AB - BACKGROUND: Nitric oxide (NO), synthesized from L-arginine by the enzyme nitric oxide synthase (NOS), seems to play an ambiguous role during tissue ischemia reperfusion (I/R) injury. This study was designed to investigate the effects of resveratrol, a polyphenolic phytoalexin, in renal ischemia reperfusion (RIR) injury in rats. METHODS: Forty-eight rats were randomized into six groups. Group 1: sham operated (C); group 2: right nephrectomy (UNI); group 3: UNI + 45 min of ischemia and 24 h of reperfusion in the contralateral kidney; group 4: UNI + RIR + L-NAME (10 mg/kg, i.p.); group 5: UNI + RIR + resveratrol (5 mg/kg, p.o.); group 6: UNI + RIR + resveratrol + L-NAME. At the end of the reperfusion period, rats were sacrificed. Thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) levels, catalase (CAT), and superoxide dismutase (SOD) activities were determined in renal tissue. Serum creatinine and blood urea nitrogen (BUN) were measured for the evaluation of renal function. Tissue and urine nitrite levels were measured to assess total nitric oxide levels. RESULTS: Ischemic control animals demonstrated severe deterioration of renal function, altered renal morphology, reduced total nitric oxide levels and a marked renal oxidative stress. CONCLUSIONS: Pretreatment of animals with resveratrol markedly attenuated renal dysfunction, morphological alterations, improved nitric oxide levels, reduced elevated TBARS levels and restored the depleted renal antioxidant enzymes, However, treatment with L-NAME attenuated this protection afforded by resveratrol indicating that resveratrol exerts its protective effect through NO release. PMID- 16314182 TI - Prolonged ventricular action potential duration due to nitric oxide release. AB - BACKGROUND: Our previous study has demonstrated that increased intracoronary perfusion leads to a flow-dependent inversion of T waves on body surface ECG. However, whether increased coronary flow influences ventricular action potential duration measured directly from myocardium is unknown. METHODS: In six pentobarbital-anesthetized sheep, fresh arterial blood was injected into the left circumflex coronary (LCX) artery at a rate of 6 and 10 mL/min, respectively, in the presence of normal coronary flow. Activation-recovery interval (ARI), an estimate of ventricular action potential duration, was measured from epicardial ECGs acquired from the LCX territory. RESULTS: The intracoronary injection prolonged ARI by an average of 21 +/- 9 and 33 +/- 14 msec, respectively. After pre-treatment with nitro-l-arginine, a nitric oxide synthase inhibitor, intracoronary injection at the rate of 6 and 10 mL/min lead to an ARI increase of 3 +/- 2 msec (p >0.05) and 11 +/- 6 msec (p <0.05) respectively. CONCLUSIONS: An increase in coronary flow prolongs ventricular action potential duration in the intact sheep heart. Nitric oxide mediates the injection-induced increase in action potential duration. PMID- 16314183 TI - Linezolid diminishes inflammatory cytokine production from human peripheral blood mononuclear cells. AB - BACKGROUND: Active peptides produced by monocytes, in response to endotoxin, initiate and maintain the acute phase of inflammatory response. Some antibiotics have been reported to have immunomodulatory effects in addition to their antimicrobial activity. We examined the effect of linezolid on cytokine synthesis. METHODS: The modulatory effects of erythromycin and linezolid were evaluated in LPS-stimulated human peripheral blood mononuclear cells (PBMCs). Blood was obtained by venipuncture from healthy donor volunteers. PBMCs were separated by Ficoll-Paque. More than 90% of the cells were monocytes as determined by esterase staining. Cells were incubated in the presence of LPS, with or without various concentrations of erythromycin and linezolid. The concentration of each cytokine was determined by ELISA with commercially available reagents. RESULTS: The two drugs suppressed significantly the synthesis of the cytokines tested in a concentration-dependent manner. CONCLUSIONS: These data indicate that antibacterial agents may modify acute phase inflammatory response through their effects on cytokines synthesis by monocytes. PMID- 16314184 TI - Altered morphology and distribution of cellular junction proteins in non-lesional psoriatic epidermis: an insight into disease severity. AB - BACKGROUND: Psoriasis affects 2.7% of the world's population. Keratinocyte proliferation outside the basal layer suggests alterations in cell-cell interactions in affected epidermis. Anomalous expression of proteins forming intercellular junctions has been reported in lesional skin of psoriatic patients. In contrast, little is known about possible alterations in psoriatic non-lesional skin. METHODS: Ten clinically diagnosed psoriasis vulgaris patients and ten controls were studied. All patients were diagnosed with active but controlled psoriatic plates (PASI 3 to 5) and had not received any systemic treatment. The mean age was 43 years for patients and 43.5 years for controls. Four-mm2 skin samples were taken from lesional and non-lesional zones in patients and from abdomen in controls. Five-mum sections were examined for integrity and structural organization by fluorescent labeling of actin filaments and nuclei. Specific antibodies were utilized to localize occludin, E-cadherin, beta-catenin, and proliferation-specific keratins in sections and epidermal sheets. Samples were also processed for immunoblotting with occludin antibody. RESULTS: Lesional and non-lesional psoriatic epidermis from all patients showed keratinocyte hyperproliferation, lack of rete ridges and dermal papillae in the dermal epidermal junction in some areas. Proteins forming tight and adherens junctions in non-lesional skin keratinocytes from two patients who during the course of the study evolved to uncontrolled disease, showed similar alterations to those observed in lesional skin of all the patients. However, the occludin isoforms expressed were apparently the same in all samples. CONCLUSIONS: Analysis of non lesional skin in psoriatic patients diagnosed with controlled disease may provide clues about incipient structural abnormalities in the pathogenesis of psoriasis, providing an early diagnostic indicator for evolution to a generalized form of the disease. PMID- 16314185 TI - Bactericidal activity of organic extracts from Flourensia cernua DC against strains of Mycobacterium tuberculosis. AB - BACKGROUND: Tuberculosis is a chronic disease caused mainly by Mycobacterium tuberculosis. The emergence of antibiotic-resistant strains of this species underscores the need for novel effective drugs against resistant mycobacteria as first-line antituberculosis medications. METHODS: Crude aqueous (obtained by decoction, in accordance with the traditional mode of preparation), methanol, acetone, and hexane extracts from aerial parts of Artemisia ludoviciana Nutt., Chenopodium ambrosioides L., Marrubium vulgare L., Mentha spicata L., and Flourensia cernua DC were assessed for their ability to either inhibit the growth of or kill M. tuberculosis strains H37Rv and CIBIN:UMF:15:99, the former being sensitive to, and the latter resistant to, streptomycin, isoniazide, rifampin, ethambutol, and pyrazinamide. These five plant species are used in Mexico to treat respiratory disorders. RESULTS: Flourensia cernua was the uniquely active plant among those evaluated. Its hexane and acetone extracts not only inhibited the growth of but killed M. tuberculosis. The hexane extract showed a minimal inhibitory concentration (MIC) of 50 and 25 microg/mL against sensitive and resistant strains, respectively; the acetone extract was active against only CIBIN:UMF:15:99 (MIC = 100 microg/mL). CONCLUSIONS: The hexane extract from F. cernua leaves could be an important source of bactericidal compounds against multidrug-resistant M. tuberculosis. PMID- 16314186 TI - Ethanolic extract of Spirulina maxima alters the vasomotor reactivity of aortic rings from obese rats. AB - BACKGROUND: Aortic rings with endothelium excised from fructose-fed obese rats develop more tension in response to phenylephrine and relax less in response to carbachol than corresponding rings from lean rats. This altered vascular reactivity is prevented when Spirulina maxima is added to the fructose-rich diet. In the present study the effects of a raw ethanolic extract of Spirulina maxima on the vasomotor responses of aorta rings from sucrose-fed obese hypertensive rats were analyzed. METHODS: The experiments were performed on aorta rings from sucrose-fed obese male rats. For each experiment, a pair of rings from the same aorta (one with intact endothelium, the other without a functional endothelium) was used. In this study we analyzed, in vitro, the effects of the ethanolic extract of Spirulina maxima on the reactivity of the aortic rings to phenylephrine and to carbachol. RESULTS: On rings with endothelium, the extract produced the following effects: a) a concentration-dependent (0.06-1.0 mg/mL) decrease of the contractile response to phenylephrine; b) a rightward shift and a decrease in maximal developed tension, of the concentration-response curve to phenylephrine; c) a concentration-dependent relaxation of phenylephrine precontracted rings. These effects persisted in the presence of indomethacin but were prevented by L-NAME. The extract had no effect on the concentration-response curve of phenylephrine-precontracted rings to carbachol. On endothelium-denuded rings the extract caused a significant rightward shift of the concentration response curve to phenylephrine without any effect on maximal tension development. CONCLUSIONS: These results suggest that, in rings from obese rats, the extract, in addition to increasing the synthesis/release of NO, also inhibits the synthesis/release of a cyclooxygenase-dependent vasoconstrictor metabolite of arachidonic acid, which is increased in obesity. PMID- 16314187 TI - Assessment of right and left ventricular perfusion in patients with right bundle branch block. AB - BACKGROUND: Patients with conduction abnormalities may have abnormal images on perfusion scintigraphy without evidence of coronary artery disease (CAD). Based on the presence of left ventricular perfusion abnormalities in left bundle branch block and taking advantage of the Tc-99m sestamibi for the assessment of myocardial perfusion of both ventricles, we aimed to evaluate right (RV) and left (LV) myocardial perfusion in patients with right bundle branch block (RBBB). METHODS: This study included 21 patients with RBBB and 21 control subjects without conduction abnormality. None had previous myocardial infarction, known CAD, left ventricular hypertrophy, LVEF <55%, congenital, pulmonary or valvular heart disease. Rest and stress SPECT images were obtained using 1-d Tc-99m sestamibi SPECT protocol. Regional perfusion of LV and RV myocardium was semiquantitatively evaluated. Also, RV/LV uptake ratio was calculated in two different ways. RESULTS: In patients with RBBB, the normality ratios of RV and LV myocardial perfusion were very high (both 90.5%) and not different from those of patients with normal intraventricular conduction. In the comparison of RV/LV uptake ratios, no significant difference was found between the RBBB and control groups. CONCLUSIONS: LV perfusion, RV perfusion or R-to-L perfusion ratio were not significantly influenced by the conduction abnormality in this selected patient population with RBBB. Tc-99m sestamibi myocardial scintigraphy could help in the exclusion of myocardial ischemia in patients with RBBB. PMID- 16314188 TI - HFE-codon 63/282 (H63D/C282Y) gene variants in Mexican Mestizos are not risk factors for leukemia. AB - BACKGROUND: In some Caucasian populations it has been found that the C282Y hemochromatosis (HFE) gene mutation is a risk factor for the development of leukemia and other malignancies. METHODS: In a group of 50 Mexican mestizo patients and 153 normal controls, the HFE gene mutations H63D and C282Y were studied by means of ARMS-PCR. RESULTS: In the group of patients with leukemia we found a heterozygote for the C282Y mutation, seven heterozygotes for the H63D mutation, a double heterozygote for the H63D / C282Y mutation and 41 normal homozygotes. These data are not different from those observed in normal controls, where the allele frequencies were 0.062 and 0.013 for the H63D and C282Y HFE gene mutations, respectively. CONCLUSIONS: These HFE gene mutations are not risk factors for the development of leukemia in Mexican mestizos. PMID- 16314189 TI - Remission of severe relapsing or persistent lupus nephritis using mycophenolate mofetil. AB - BACKGROUND: The therapy of severe lupus nephritis (LN) consists of high-dose steroids and immunosuppressive agents, usually cyclophosphamide. Although effective in up to 90% of cases, this approach leads to undesirable complications in many cases. In recent years, mycophenolate mofetil (MMF), an immunosuppressive drug used in transplantation regimes, seems to be effective in selected cases of lupus nephritis. METHODS: In this report we present the results using MMF in seven cases of LN. MMF stabilized renal function, controlled extrarenal disease activity and led to less steroid dosing. RESULTS: Three patients achieved complete remission, two partial remissions and two failed, although these two patients were the ones with the highest chronicity indices in the renal biopsies. However, they were able to maintain stable renal function for more than one year with tolerable side effects. In two more patients the latter were mild and did not require either hospitalization or intense therapy. CONCLUSIONS: MMF can be an option for selected cases with severe relapsing or persistent LN and can lead to induction of remission. This observation needs to be expanded. Larger randomized controlled studies are needed to evaluate its indication in earlier cases as induction of de novo disease and/or effective remission maintenance. PMID- 16314190 TI - Thyroid hormone and insulin-like growth factor-I in patients with multiple myeloma treated with melphalan and prednisone. AB - BACKGROUND: The relationship between endocrine regulation and immune system has recently become the subject of intense investigations. The aim of this study was the comparative assessment of serum levels of selected hormones and insulin-like growth factor-I (IGF-I) in patients with multiple myeloma (MM) during applied therapy. METHODS: The levels of prolactin, hGH, TSH, fT3, fT4 and IGF-I in serum of 13 untreated patients with MM and in 16 healthy controls were determined. The patients were treated in cyclic courses with melphalan plus prednisone, and investigations were carried out in the first four courses of this therapy. The results were compared in the following manner: (1) at entry between studied MM group and healthy subjects, and (2) during the therapy intragroup-intracyclic comparisons were made in paired serum samples collected from patients before and after every therapeutic course. RESULTS: At entry, significantly lower levels of TSH and fT3 were obtained in MM patients. The means remained within low normal reference range. Slightly increasing levels of TSH and fT3 during treatment with lower concentrations of these hormones after every therapeutic course and a statistically significant difference of fT3 level in the fourth therapy course were revealed. The levels of fT4 were within the normal reference values and showed a tendency to decrease during therapy with significant differences in the first therapeutic course. After the third and the fourth therapy courses, concentrations of IGF-I were statistically significantly higher than initially. CONCLUSIONS: Euthyroid sick syndrome can exist in MM patients, and the therapy with melphalan plus prednisone is accompanied by slightly expressed serum changes of thyroid hormone concentrations and IGF-I levels. PMID- 16314191 TI - T2 hyperintensities in bipolar patients and their healthy siblings. AB - BACKGROUND: This study investigates the frequency of hyperintensities, which are common in bipolar disorder, in sufferers' siblings who are free of bipolar disorder and to ascertain whether these lesions reflect any familial characteristics. It offers an assessment of the relationship between these lesions and clinical characteristics. METHODS: The study group consisted of 12 patients with bipolar disorder, their siblings who had no history of mental disorder, and a matched control group. All three groups were assessed using SCID I (Structured Clinical Interview for DSM-IV Axis I Disorders) for diagnosis, and MRI (magnetic resonance imaging) for signal hyperintensity. RESULTS: No periventricular white matter lesions were encountered in any groups. It was found that in the patients and siblings, the location was in the right cerebral hemisphere in the deep white matter, whereas it was in the left cerebral hemisphere for the control group. Only in the patient group were lesions detected in subcortical white matter in the right cerebral hemisphere. The lesions were localized in the fronto-parietal area. Considering the brain as a whole, more hyperintensities were detected in the patient group compared to the other two groups. The onset of the disorder with a manic episode was increasing probably in the presence of hyperintensities. CONCLUSIONS: Despite being alike in terms of age, more hyperintensities were detected in the patient group than in the siblings and control groups. Contrary to the control group, however, hyperintensities were localized in the right cerebral hemisphere in both the patients and the siblings groups. Hyperintensities seen in the left cerebral hemisphere yield a nonspecific impression. Siblings who have hyperintensities in the right cerebral hemisphere should be followed up and investigated with regard to bipolar disorder. PMID- 16314192 TI - Effects of two common polymorphisms of peroxisome proliferator-activated receptor gamma gene on metabolic syndrome. AB - BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)gamma is involved mainly in adipocyte differentiation and has been suggested to play an important role in the pathogenesis of insulin resistance and atherosclerosis. We investigated the frequencies of two common polymorphisms of PPARgamma gene, exon 6 C-->T substitution and exon B Pro12Ala in healthy subjects and analyzed the correlations between the different genotypes and insulin resistance, metabolic syndrome and cardiovascular risk factors. METHODS: Anthropometric measurements, fasting glucose, insulin and lipid profiles were measured in 253 Korean females. Homeostatic model assessments and quantitative insulin sensitivity check indices were calculated. Metabolic syndrome was diagnosed according to the NCEP-ATP III guidelines and the Western Pacific Region of WHO for obesity criteria for waist circumference. Polymerase chain reaction (PCR)-restriction fragment-length polymorphism and real-time PCR were performed for genotyping of the DNAs. RESULTS: For C161T polymorphism, allele frequencies were 0.804 and 0.196 for T allele, and 0.947 for proline and 0.053 for alanine. There was no Ala12Ala homozygote in the population. No differences were seen in the mean values of age, body mass index (BMI), blood pressure, fasting blood glucose level, fasting insulin levels, HOMA and QUICKI among different genotypes when analyzed as a whole, except that subjects with Pro12Ala had significantly higher body weight than those with Pro12Pro genotype. However, mean BMI, percent body fat and weight showed significant differences between genotypes in younger age group (< or =50 years). Although overall prevalence of metabolic syndrome had no association with the genotypes, the prevalence of decreased high-density lipoprotein cholesterol component was lower in those with the T allele than in those with the CC genotype. There was no association of the genotypes with glucose tolerance status. When the subjects were divided into four groups according to the combination of the genetic alleles of the two polymorphisms, subjects having Pro12Ala and T allele, simultaneously, showed significantly higher mean weight than those without Ala allele. Pro12Ala polymorphism seems to affect body weight, similar to the previous studies, and the effect was potentiated with the presence of T allele of C161T polymorphism. CONCLUSIONS: Although either polymorphism failed to show significant association with insulin resistance, the fact that the prevalence of decreased HDL-C was lower in those with the T allele of C161T polymorphism suggests that this polymorphism might have a protective effect on atherosclerotic lipid profiles, which needs further investigation. PMID- 16314193 TI - Phenotypes of isolates of Pseudomonas aeruginosa in a diabetes care center. AB - BACKGROUND: Pseudomonas aeruginosa is an invasive organism that frequently causes severe tissue damage in diabetic foot ulcers. A major problem in P. aeruginosa infection may be that this pathogen exhibits a high degree of resistance to a broad spectrum of antibiotics. Some researchers feel that P. aeruginosa is a homogeneous species, whereas others have suggested that they are panmictic. Here we characterized P. aeruginosa populations isolated from diabetic foot ulcer and from hospital environment specimens, both from a tertiary diabetes care center in Chennai, India. METHODS: Phenotypic methods like antibiotic susceptibility determinations using Kirby-Bauer's disc diffusion test and minimum inhibitory concentration (MIC) as well as outer membrane protein SDS-PAGE analysis of P. aeruginosa were performed. RESULTS: Twenty three isolates (29.8%) of P. aeruginosa from 77 diabetic foot ulcers and two environmental isolates (13.3%) from 15 different hospital fomites were detected. Both environmental isolates were sensitive to antibiotics than those isolated from clinical specimens by Kirby-Bauer's disk-diffusion method, which correlated the resistance levels by MIC determination. Outer membrane proteins (OMP) corresponding to 21, 23, 43, 46, 50, and 70 kDa were detected. CONCLUSIONS: The study is captivative as the resistance in P. aeruginosa from diabetic foot ulcers seems very common and because all the isolates were resistant to at least one or more antibiotics tested. Disk-diffusion and MIC results shows that piperacillin, amikacin and imipenem retain high levels of antipseudomonal activities and amikacin two times more active than the aforementioned antibiotics to enable itself as a potent antipseudomonal agent in diabetic foot infections. The OMP profile has revealed that clinical isolates were different from hospital environment isolates, which suggests that the origin of infections by P. aeruginosa is mainly due to growth of bacterial strains acquired by patients prior to hospital admission. PMID- 16314194 TI - Genetic heterogeneity of autosomal dominant hypercholesterolemia in Mexico. AB - BACKGROUND: Familial hypercholesterolemia (FH) and familial defective apolipoprotein B-100 (FDB) are relatively common lipid disorders caused by mutations of the low-density lipoprotein receptor (LDLR) and apolipoprotein B (apoB) genes, respectively. A third locus on chromosome 1p34.1-p32 was recently linked to FH and the responsible gene has been identified [protein convertase subtilisin/kexin type 9 (PCSK9)]. METHODS: We assessed the contribution of the LDLR, apoB, and PCSK9 genes as cause of FH in Mexico. Forty six unrelated probands, as well as 68 affected and 60 healthy relatives, were included. RESULTS: All index cases were diagnosed as having heterozygous autosomal dominant FH. Seventeen of the 46 index cases had LDLR gene mutations, four of which were novel (Fs92ter108, C268R, Q718X, and Fs736ter743); and only one patient had an apoB mutation (R3500Q). We sequenced the PCSK9 gene in the remainder of the 28 probands with no identified LDLR or APOB gene defects; however, no PCSK9 mutations were found, including one large kindred with positive linkage to the 1p34.1-32 locus (multipoint LOD score of 3.3) and two small pedigrees. Linkage was excluded from these three loci in at least four kindreds suggesting that other yet uncharacterized genes are involved. CONCLUSIONS: Our results underline substantial genetic heterogeneity for FH in the Mexican population. PMID- 16314195 TI - Elevation of hyaluronidase-1 and soluble intercellular adhesion molecule-1 helps select bladder cancer patients at risk of invasion. AB - BACKGROUND: The identification of accurate bladder tumor marker/tests could improve diagnosis, recurrence monitoring and treatment in patients with bladder cancer. This study evaluates the potential usefulness of hyaluronidase-1 (HYAL-1) and soluble intercellular adhesion molecule-1 (sICAM-1) in Egyptian bladder cancer patients. METHODS: A total of 210 tissue, serum and urine specimens were used to investigate HYAL-1 and sICAM-1 expression by reverse transcriptase polymerase chain reaction (RT-PCR) and enzyme immunoassay (EIA) techniques, respectively. RESULTS: The detection of urinary and tissue HYAL-1 mRNA as well as urinary and circulating sICAM-1 were higher in the malignant than the control group (p <0.0001). In the cancer group, both HYAL-1 and sICAM-1 were significantly correlated with each other and with the patient's age. Also, they were increased in advanced and poorly differentiated tumors (p <0.01). Patients with positive HYAL-1 had elevated sICAM-1 in their serum and urine (p <0.05). Although sometimes statistically insignificant, both markers tended to increase in lymph node and cytology positive than the corresponding negative subgroups. The levels of sICAM-1 (urinary and circulating) were elevated in relation to schistosomal infection (p <0.01). Using receiver operating characteristic curve, the best cut-off values for urinary and circulating sICAM-1 were 110 ng/mg creatinine and 130 ng/mL, respectively. Sensitivity, specificity and efficiency were >80%. CONCLUSIONS: Serial monitoring of sICAM-1 (in urine and/or serum) is potentially recommended for selecting patients who are at risk of tumor invasion. PMID- 16314196 TI - Placebo or trimetazidine (99m)Tc tetrofosmin myocardial SPECT and low-dose dobutamine echocardiography in hibernating myocardium. AB - BACKGROUND: Trimetazidine (TMZ) improves (99m)Tc sestamibi uptake in myocardial single photon emission tomography (SPECT). This study compared TMZ (99m)Tc tetrofosmin SPECT and low-dose dobutamine echocardiography (LDDE) as predictors of functional recovery of hibernating myocardium after coronary revascularization. METHODS: Thirty-one patients with prior myocardial infarction and left ventricular dysfunction underwent coronary angiography, LDDE, placebo SPECT and TMZ SPECT. Echocardiographic follow-up was obtained at 2/6 months; the clinical follow-up lasted 2 years. RESULTS: Twenty-three (74.2%) patients (195 dysfunctioning left ventricular segments) were revascularized. TMZ improved (99m)Tc tetrofosmin uptake (p = 0.0001) as well as LVEF at gated SPECT (p = 0.04). At 2-months, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated (LDDE 71.9, 78.7, 71, 79.5; placebo SPECT 66.2, 75.6, 65.4, 76.3; TMZ SPECT 79.2, 67.7, 61.6, 83.3, respectively). The specificity improved in placebo and TMZ SPECT (82.1 and 78.7%) at 6-months follow-up. Two patients (2/23) with hibernating myocardium treated with revascularization and three (3/4) treated medically died a cardiac death (p = 0.0016, log rank 12.89). None (0/4) without viability died during the 2-year follow-up (p = 0.6, log rank 0.28). CONCLUSIONS: The addition of TMZ to (99m)Tc tetrofosmin SPECT improved diagnostic accuracy. The importance of hibernating myocardium revascularization was confirmed. PMID- 16314197 TI - Diagnostic utility of invasive tests and serology for the diagnosis of Helicobacter pylori infection in different clinical presentations. AB - BACKGROUND: Invasive and noninvasive tests are used for the diagnosis of Helicobacter pylori infection. The aim of this study was to determine the diagnostic utility of rapid urease test (RUT), culture, histology and serology for the diagnosis of H. pylori in patients with different clinical presentations. METHODS: We studied 527 consecutive patients (mean age, 52.5 years; F:M, 1.3; age range 15-89 years) enrolled at the Hospital Universitario, Universidad Autonoma de Nuevo Leon. Patients had gastric cancer (GC, 9.1%), non-ulcer dyspepsia (NUD, 81.4%), or peptic ulcer disease (PUD, 9.1%). The infection by H. pylori was determined by histology, rapid urease test, culture, and serology. Patients were determined as infected with H. pylori if at least a) two invasive tests were positive and b) two tests were positive (invasive or non-invasive). Diagnostic utility was calculated for each assay. RESULTS: Prevalence of infection in the whole studied population was 50.9%. In NUD patients the prevalence was 51.3%, in PUD patients 58.3%, and in GC patients 39.6%. When we used the first diagnostic criteria, for the whole studied population, the RUT was the most reliable test, followed by the culture. Histology had the best sensitivity for the whole studied population and NUD patients and RUT had the best sensitivity value for the GC patients. In the whole studied population, NUD and GC patients, RUT and culture had the best specificity, accuracy and PPV. For PUD patients, serology had the best performance. When we used the second diagnostic criteria, histology and serology had a better performance compared with the results obtained with the first diagnostic criteria. CONCLUSIONS: Diagnostic utility of the tests varies according to the clinical presentations, which should be considered in the selection of the diagnostic test for the detection of H. pylori. PMID- 16314198 TI - Hypertriglyceridemia and hypercholesterolemia in human immunodeficiency virus-1 infected children treated with protease inhibitors. AB - BACKGROUND: Adverse effects associated with highly active antiretroviral therapy (HAART), particularly protease inhibitors (PIs), have been identified in adult and pediatric patients. In this study, we monitored, for cholesterol and triglyceride levels, a cohort of HIV-1-infected children receiving a HAART regimen. METHODS: HIV-infected patients <17 years old belonging to a cohort that has been followed since 1997 were enrolled in the study. Patients were receiving either a three- or four-drug antiretroviral regimen that included two nucleoside reverse transcriptase inhibitors (lamivudine and zidovudine) combined with one or two PIs (ritonavir and/or saquinavir). Cholesterol and triglyceride levels were measured on fasting serum samples drawn at the time of enrollment and every 3 months thereafter. Clinical evaluation was performed on a monthly basis. RESULTS: Twenty four patients were included. Median age at HIV infection diagnosis was 15 months. Twenty one patients received a four-drug antiretroviral regimen, while three patients received ritonavir plus zidovudine and lamivudine. Median follow up was 27 months; 62.5% of patients had hypercholesterolemia and 79.2% had hypertriglyceridemia, most typically after 15 months of treatment. None of the patients had physical changes in body fat distribution suggesting lipodystrophy. CONCLUSIONS: Hyperlipidemia is a frequent complication in HIV-1-infected children undergoing antiretroviral treatment that includes PIs. Additional studies with larger cohorts and a longer follow-up are needed to propose a rationale and alternatives for patients who develop dyslipidemia while receiving PIs. PMID- 16314199 TI - Depression, anxiety, health-related quality of life, and disability in patients with overt and subclinical thyroid dysfunction. AB - BACKGROUND: Thyroid dysfunction may impair the quality of life (QoL) and may cause psychological symptoms. The aim of this study is to investigate prospectively the effects of thyroid dysfunction on quality of life, levels of depression/anxiety and the changes in these variables after treatment. METHODS: A total of 160 subjects, consisting of an overt hypothyroidism group (n = 33), a subclinical hypothyroidism group (n = 43), an overt hyperthyroidism group (n = 51), a subclinical hyperthyroidism group (n = 13), and a healthy control group (n = 20) were included in the study. All groups were evaluated with the Hamilton Depression Rating Scale (HDRS), Hamilton Anxiety Rating Scale (HARS), Short Form 36 (SF-36), and Brief Disability Questionnaire (BDQ). Health-related quality of life (HRQL) was measured by SF-36. RESULTS: Anxiety and depressive symptoms were more severe in patients with overt hypo- and hyperthyroidism (p <0.001). The QoL was worse in overt or subclinical hyper- and hypothyroidism groups than in the control group [p = 0.013 for physical composite score (PCS); p = 0.002 for mental composite score (MCS)]. Psychological symptoms and QoL were improved in overt and subclinical hypothyroidism and overt hyperthyroidism groups as a result of the treatment. The overt hyper- and hypothyroidism groups showed more improvement than the subclinical groups. CONCLUSIONS: This study suggests that restoration of euthyroidism is accompanied by improvement on QoL and psychological symptoms in all groups except the subclinical hyperthyroidism group. Controlled, randomized studies in larger groups are, however, necessary. PMID- 16314200 TI - Lowered criterion for normal fasting plasma glucose: impact on the detection of impaired glucose tolerance and metabolic syndrome. AB - BACKGROUND: Recently, the Expert Committee of the American Diabetes Association proposed to decrease the lower limit for normal fasting plasma glucose (FPG) levels to 5.6 mmol/L. In this regard, the aim of this study was to determine the effect of lowering the criterion for normal FPG on the identification of subjects with impaired glucose tolerance (IGT) and metabolic syndrome (MS). METHODS: We carried out a cross-sectional analysis of a population-based study from Durango, Mexico. This study enrolled 844 apparently healthy men and nonpregnant women aged 34 to 64 years. Diagnosis of IGT was based on 2-h post-load plasma glucose concentration > or =7.8 mmol/L and <11.1 mmol/L, and MS by the definition of National Cholesterol Education Program. RESULTS: The diagnosis of IGT and MS was established in 161 (19.1%) and 190 (22.5%) individuals, respectively. Lowering the cut-off point of FPG from 6.1 to 5.6 mmol/L identified more subjects with IGT, 53/161 (32.9%) vs. 132/161 (82.0%) (p <0.0001). It also identified more subjects with MS, from 88/190 (46.3%) vs. 169/190 (88.9%) (p = 0.0001). Lowering of the FPG cut-off point to 5.6 mmol/L improved the positive likelihood ratio for identifying subjects with IGT from 1.91 to 2.55 and the negative likelihood ratio from 0.81 to 0.27. Lowering the FPG cut-off point to 5.6 mmol/L did not significantly change the positive likelihood ratio for identifying MS from 3.65 to 3.18 and improved the negative likelihood ratio from 0.61 to 0.15. CONCLUSIONS: Lowering the cut-off point of IFG to 5.6 mmol/L improves the predictive discrimination of IFG for identifying subjects with IGT or MS. PMID- 16314201 TI - Interictal regional cerebral blood flow and electrical source analysis in patients with complex partial seizures. AB - BACKGROUND: Recently, new methods of EEG source analysis have been developed. Dipole modeling and brain distributed source analysis localize in three dimensions the origin of the electrical source of spikes registered in EEG. With single photon emission computed tomography (SPECT), it is possible to detect hypo and hyperperfusion zones. Our aim in this study is to compare the regions where the electrical sources are detected, with the hypoperfusion regions in patients with complex partial seizures (CPS). METHODS: The concordance of localization with dipole analysis, brain distributed source analysis and regional cerebral flow blood in patients with CPS was studied. The hypoperfusion zones detected with interictal SPECT were compared with electrical sources localized with brain electric source analysis (BESA) and brain distributed source analysis with variable resolution electromagnetic tomography (VARETA). RESULTS: Hypoperfusion zones were found to localize with the origin of dipoles in 18 cases (90%), between lobes in 17 (85%) and between mesial or lateral regions in the temporal lobe in 12 cases (60%). With VARETA, agreement between side of hypoperfusion and electrical current source localization was found in 18/20 cases (90%), with lobes in 17 (85%) and with mesial or lateral regions of the temporal lobe only in 2 cases (10%). CONCLUSIONS: Hypoperfusion zones in interictal SPECT of patients with CPS are in agreement with the origin of dipoles in 85% of the cases, but in specifics zones of temporal lobe the agreement falls to 60%. The concordance of hypoperfusion zones was better with dipole analysis than with VARETA. PMID- 16314202 TI - GNB3 C825T and ACE I/D polymorphisms on the sodium-proton exchanger and the prevalence of essential hypertension in males. AB - BACKGROUND: The aim of the study was to verify the hypothesis if the interaction between the G protein beta3 subunit (GNB3) C825T polymorphism and ACE I/D polymorphism could lead to the disclosure of increased activity of sodium-proton exchanger and hypertension. METHODS: The study included 44 male patients, median age: 40 years. Patients were divided into two groups: 26 patients with essential hypertension (EH), and 18 subjects in the normotensive group (C). RESULTS: CT + TT genotypes of GNB3 predominated in patients with hypertension (65%) compared to normotensive patients (12%) (p <0.01). No significant differences were observed in the frequency of ACE gene polymorphisms between the examined groups. Significantly higher activity of erythrocyte NHE in patients with EH was observed: median 8.83 (interquartile range 4.27) mmol/l RBC/h, compared to C: median 6.18 (2.80) mmol/l RBC/h, p <0.001. Multiple logistic regression analysis showed that the presence of the T allele increased the risk of hypertension 16 fold (p <0.01) and higher erythrocyte NHE activity 2-fold per each unit of activity (p <0.01). DD genotype of ACE polymorphism did not increase the risk of hypertension. No significant interaction of the influence of GNB3 T allele and ACE DD genotype on the risk of hypertension was observed. In multiple linear regression analysis, none of the examined genotypes and their interactions influenced NHE activity. CONCLUSIONS: The presence of the T allele of GNB3 polymorphism and increased activity of erythrocyte NHE independently of ACE genotype increase the risk of hypertension. PMID- 16314203 TI - Mangifera indica L. (Vimang) protection against serum oxidative stress in elderly humans. AB - BACKGROUND: We searched for the protective effect of a natural extract from stem bark of Mangifera indica L. extract (Vimang) on age-related oxidative stress. METHODS: Healthy subjects were classified in two groups, elderly (>65 years) and young group (<26 years). The elderly group received a daily dose of 900 mg of extract (three coated Vimang tablets, 300 mg each, before meals) for 60 days. Serum concentration of lipid peroxides, serum peroxidation potential, extracellular superoxide dismutase activity (EC-SOD), glutathione status (GSH, GSSG, GSSG/GSH ratio)) and total antioxidant status (TAS) were determined before (both experimental groups) and 15, 30, and 60 days after treatment (only elderly group). We confirmed the existence of an age-associated oxidative stress in human serum as documented by an age-related increase in serum lipoperoxides and GSSG and a decrease in serum antioxidant capacity and EC-SOD activity. RESULTS: Vimang tablet supplementation increased EC-SOD activity (p <0.01) and serum TAS (p <0.01). It also decreased serum thiobarbituric reactive substances (p <0.01) and GSSG levels (p <0.05). We suggested that the antioxidant components of the extract could have been utilized by the cells (especially blood and endothelial cells), sparing the intra- and extracellular antioxidant system and increasing serum peroxil scavenging capacity, thus preventing age-associated increase in GSH oxidation and lipoperoxidation. CONCLUSIONS: Vimang tablets prevent age associated oxidative stress in elderly humans, which could retard the onset of age-associated disease, improving the quality of life for elderly persons. PMID- 16314204 TI - Obesity among children residing in Mexico City and its impact on lung function: a comparison with Mexican-Americans. AB - BACKGROUND: The objective of the study was to describe the prevalence of obesity among Mexican children and its impact on ventilatory lung function. METHODS: We studied cross-sectionally 6784 students between 8 and 20 years of age attending schools located <2 km away from ten air pollution monitors located throughout metropolitan Mexico City. The comparison group was made up of 1924 Mexican Americans of the same age, studied during the NHANES-III examination, which included information on spirometry and body mass index (BMI). RESULTS: Of all our subjects, 9.7% had a BMI >95(th) percentile of CDC growth charts (compared to 15.1% in Mexican-Americans) and 6.6% fulfilled the obesity criteria of the International Obesity Task Force (vs. 12.2%). Obesity was related to male gender, asthma and passive smoking. At the same height and gender, lung function was higher in Mexicans than in Mexican-Americans, perhaps due to altitude. In children 8-11 years of age, lung function increased in heavier subjects but, in older children and youths, function reached a plateau and decreased among children with highest BMI (inverted U pattern, seen in adults). CONCLUSIONS: Obesity is higher in Mexican-American children and youths than in Mexicans. Spirometric function is affected adversely by obesity, especially in young people. PMID- 16314205 TI - Frequency of HPA-15a and HPA-15b (Gov a/b) human platelet alloantigens in the Croatian population. AB - BACKGROUND: Human platelet antigen (HPA) genotyping is important for epidemiological studies because the prevalence of particular HPA allotypes differs among various populations and plays a major role in the occurrence of HPA alloimmunization. In Caucasians, antibodies to HPA-1a are the most important causes of neonatal alloimmune thrombocytopenia (NATP). Recent studies suggest that anti-HPA 15a/15b (Gov b, Gov a) might be the most likely candidate antibodies following anti-HPA-1a in inducing NATP. METHODS: In the present study, HPA-15 system genotype was determined by PCR-SSP method in 279 unrelated subjects from the Croatian population, yielding an HPA-15a and HPA-15b frequency of 0.53 and 0.47, respectively. RESULTS: Retrograde testing for the presence of anti-HPA 15 antibodies by use of MAIPA in 39 frozen serum samples from serologically negative cases of clinically suspect NATP produced negative results. CONCLUSION: The clinical role of anti-HPA-15 alloantibodies was unable to be confirmed. PMID- 16314206 TI - Japanese guidelines-based management of lipid levels in a hypercholesterolemia education class. AB - The "Japan Atherosclerosis Society (JAS) Guidelines for Diagnosis and Treatment of Atherosclerotic Cardiovascular Diseases" proposed the recommended lipoprotein levels in each patient according to some categories classified by the risk factor numbers. Because the categorization is somewhat complex for some patients, we redeveloped the JAS guidelines-based survey into a simple form to easily assess each lipoprotein target level. Forty participants in the hypercholesterolemia education classes were divided into the ordinary approach group (n = 20; mean age 59 years) and the intervention group using the survey (age 58 years). When participants assessed the class by 100-point scales, the intervention group had significantly more understanding levels than the ordinary approach group (86.7 vs. 75.1 points). Our results suggested that the simplified JAS guidelines-based survey system was effectively applicable to transmitting information about lipoprotein target levels among hypercholesterolemic subjects as an educational tool. PMID- 16314207 TI - Familial mediterranean fever-related nephrotic syndrome and successful full-term pregnancy. AB - Familial Mediterranean fever (FMF) is a systemic disease with an autosomal recessive inheritance. The most serious complication of FMF is renal amyloidosis. Pregnancy may adversely affect renal function in FMF patients with amyloidosis and nephrotic syndrome. A 20-year-old woman with FMF related nephrotic syndrome became pregnant while receiving colchicine therapy. Colchicine treatment was continued during pregnancy with close observation. She gave birth to a 2400 g healthy female newborn at the 38th week of gestation. Pregnancy and neonatal outcome were uneventful. It is advisable to continue colchicine treatment during conception and pregnancy in FMF patients with amyloid related nephrotic syndrome. Colchicine treatment with bed rest, protein reinforcement, acetylsalicylic acid administration and close follow-up may improve the outcome of pregnancy in FMF patients. PMID- 16314208 TI - Hippocrates and genomic medicine. PMID- 16314209 TI - Pulse wave velocity measurements on life-style modifications in a hypercholesterolemia education class. PMID- 16314210 TI - Socio-psychological activities associated with laughter in older Japanese females. PMID- 16314211 TI - Using exterior building surface films to assess human exposure and health risks from PCDD/Fs in New York City, USA, after the World Trade Center attacks. AB - Concentrations of tetra- through octa-chlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) were determined in exterior window films from Manhattan and Brooklyn in New York City (NYC), USA, 6 weeks after the World Trade Center (WTC) attacks of 11 September 2001. High concentrations of the 2,3,7,8 substituted congeners (P(2378)CDD/Fs) were observed, at levels up to 6600 pg-TEQ g(-1) nearest the WTC site. An equilibrium partitioning model was developed to reconstruct total gas + particle-phase atmospheric concentrations of P(2378)CDD/Fs at each site. The reconstructed atmospheric and window film concentrations were subsequently used in a preliminary human health risk assessment to estimate the potential cancer and non-cancer risks posed to residents of lower Manhattan from these contaminants over the 6 week exposure period between the WTC attacks and sampling dates. Residents of lower Manhattan appear to have a slightly elevated cancer risk (up to 1.6% increase over background) and increased P(2378)CDD/F body burden (up to 8.0% increase over background) because of above-background exposure to high concentrations of P(2378)CDD/Fs produced from the WTC attacks during the short period between 11 September 2001, and window film sampling 6 weeks later. PMID- 16314213 TI - The scientific context and basis of the pharmacologic management of dry eyes. AB - The ability to diagnose and treat dry eye has improved dramatically over the last decade as understanding of disease mechanisms and pathology has improved. The ability to screen patients for dry eye before ophthalmic plastics procedures has improved, and the ability to manage and treat dry eye in patients after surgery also has improved. PMID- 16314214 TI - Ocular allergy: diagnosis and treatment. AB - Ocular allergy is a common hypersensitivity disorder that affects 15%-20% of the population in developed nations. This disorder can be divided into several categories: seasonal and perennial allergic conjunctivitis, atopic keratoconjunctivitis, vernal keratoconjunctivitis, and drug-induced allergic conjunctivitis. Giant papillary conjunctivitis often is included among these classifications of allergy; however, it is not a true allergy, but rather the result of a chronic mechanical irritation. This article reviews these subtypes of ocular allergy, with particular emphasis on the pathophysiology, diagnosis, and treatment of seasonal and perennial allergic conjunctivitis. These are the most common varieties, representing more than 95% of the ocular allergy cases in the United States. PMID- 16314215 TI - New antimicrobials in ophthalmology. AB - This article discusses the newest antibiotics, antivirals, and antifungals and their application in ophthalmology. The literature is reviewed to present the latest research on topical antibiotics, with particular attention to the newest classes of fluoroquinolones, new oral and topical antifungals, and new oral antivirals. Data comparing in vitro susceptibilities and minimum inhibitory concentrations of the antibiotics and antifungals are presented in tabular form. PMID- 16314216 TI - Immunosuppressive agents and nonsteroidal anti-inflammatory drugs for ocular immune and inflammatory disorders. AB - We now have at our disposal several nonsteroidal immunosuppressive and anti inflammatory agents that may be used in addition to or instead of corticosteroids to treat ocular diseases. This article discusses some of the nonsteroidal immunosuppressive and anti-inflammatory agents available to the ophthalmologist. PMID- 16314217 TI - Ocular hypotensive medications for the treatment of glaucoma. AB - The past 10 years have seen dramatic changes in the pharmacologic treatment of glaucoma. The treatment of glaucoma has changed from surgical to primarily medical. These treatments have improved the effectiveness and safety of lowering the intraocular pressure to prevent progressive vision loss and blindness, but no cure for glaucoma or reversal of blindness is available yet. Research is progressing and can hopefully achieve these final goals sometime in the future. PMID- 16314218 TI - Current and future prospects for the prevention of ocular fibrosis. AB - Significant advances have been made in developing new treatments and refining existing treatments for the prevention of scarring after disease, trauma, or surgical prevention. The advent of new technologies in addition to traditional chemical drugs such as dendrimers, antibodies, aptamers, ribozymes, gene therapy with viral vectors, and RNA interference, opens the door to a whole new generation of therapies to prevent fibrosis in the eye. The ability to control fibrotic processes in the eye offers many tantalizing prospects, including prevention of corneal blindness from scarring to "20/5 vision" with perfect corneal wound healing after wavefront refractive surgery prevention of PCO to fully accommodative lens implants, 100% success of glaucoma surgery with pressure at approximately 10 mmHg associated with < 5% progression over a decade, to no failure of retinal detachment surgery and minimal visual loss from AMD. Finally, most exciting is the prospect that neutralizing the fibrotic response to disease and injury will allow us to revert to the "fetal" mode when regeneration is the normal process, such as shown in the recent report that demonstrated that induction of bcl-2 gene expression together with downregulation of gliosis results in axonal regeneration in mice. PMID- 16314219 TI - Anti-macular degeneration agents. AB - Anti-VEGF therapy is a promising new avenue for the treatment of ocular neovascular diseases. Early preclinical data and recent clinical data support the efficacy and safety of several novel anti-VEGF for NVAMD. Whether these novel biologics are used on their own, in combination with previously available therapies, or with newly developing therapies, they represent a new avenue in treatment. These agents are highly selective in their targeted approaches, and when administered appropriately , offer treatment with minimal damage to retinal tissue. In the future, biotherapeutic agents will certainly play a powerful role in the treatment of human choroidal neovascular membrane formation. PMID- 16314220 TI - Systemic reactions associated with ophthalmic medications. AB - Although most medications for ophthalmic disease are administered locally, systemic absorption occurs as the drug passes into the nasopharynx by way of the nasolacrimal duct and may lead to a significant exposure. Knowledge of the mechanism of action of the drug, extent of systemic exposure, and its corresponding risks are important factors that must be considered before prescription of any ophthalmic agent. The relative risk in turn depends on the agent prescribed, plasma levels achieved, and individual susceptibility factors. Judicious use of pharmacotherapy in the management of ophthalmic disease should minimize the risk-benefit ratio. PMID- 16314221 TI - Neuroprotection and regeneration in glaucoma. AB - Glaucoma is the most common optic neuropathy. Preventing visual loss is being studied with neuroprotective therapies. Visual restoration requires a restoration of retinal ganglion cells and their axons, ie, neurorepair and neuroregeneration. This review surveys recent developments in neuroprotection and regeneration from the standpoint of eventual applicability to treatment of patients who have glaucoma. PMID- 16314222 TI - Complementary therapy for the treatment of glaucoma: a perspective. AB - Although neuroprotective strategies and pharmaceutical agents have been initiated in the treatment of numerous disorders of the central and peripheral nervous systems, including trauma, epilepsy, stroke, Huntington disease, amyotrophic lateral sclerosis,and AIDS dementia, none have yet been applied to the treatment of glaucoma. A prospective, placebo-controlled, multi-institutional trial of memantine is underway. One would not expect the treatment modalities that form the bases of nonpharmaceutical, traditional medical systems to be used to lower IOP. Glaucoma was unknown when these medicinal treatments were developed over the centuries. Their primary use is in improving the cardiovascular and immune systems and in what is now called neuro-protection. Rather than single compounds that target a specific receptor and have demarcated side effects in other systems, plant products are a blend of many compounds and, according to those most versed in them, they achieve a balanced therapy, helping in specific symptomatic complexes while reducing side effects through ameliorating effects in other areas. It is not insignificant that, now that the rain forests are rapidly dwindling, together with their inhabitants and the knowledge of medicinal plants (especially in South America), the pharmaceutical companies are spending large amounts of money in a sudden, almost frantic attempt to gather the knowledge about rainforest plants before all has been completely lost. Proof of effects clinically in a chronic disease such as glaucoma remains largely lacking, and controlled trials are unlikely to be initiated, except perhaps through the National Institutes of Health, because these compounds have been in the public domain for many years. Perhaps those as yet unknown or un-recorded are patentable and perhaps these include drugs known only to small surviving communities of hunter-gatherers, which explains the pharmaceutical interest in these areas. When more accurate and rapid means of assessment of progression of glaucomatous damage than perimetry and optic nerve head photography are eventually developed and trials can be reduced in time, number of subjects, or even the use of nonhuman subjects for the bulk of studies, studies could be done for verification of effect of various compounds and also comparative studies. At the present time, GBE is the best documented of all the complementary medicinal agents and seems to have the greatest potential value. Ginkgo biloba extract has numerous properties that theoretically should be beneficial in treating non-IOP-dependent mechanisms in glaucoma. Its multi-ple beneficial actions, including increased ocular blood flow, antioxidant activity, platelet activating factor inhibitory activity, nitric oxide inhibition, and neuroprotective activity, combine to suggest that GBE could prove to be of major therapeutic value in the treatment of glaucoma. PMID- 16314223 TI - Compliance with medicine. AB - Compliance with medication regimens is critical in assessing the effectiveness of treatments. As new drugs are approved for marketing and prescribed to diverse patient populations, the FDA now recommends further observational studies to continuously monitor unforeseen side effects or efficacy. More research is needed to develop valid and reliable tools to assess adherence of patients to treatment recommendations and adherence of patients to treatment guidelines, when applicable. Information on adherence is a relevant to physicians and their patients as it is to insurers and payers, who need to implement cost-effective disease management programs. This review also has highlighted examples in the adherence literature specific to glaucoma medications and based on longitudinal survival analyses of claims data. Such results may be best complemented by primary, survey-based data collected from patients in observational studies. PMID- 16314224 TI - Diet and cardiovascular disease: beyond cholesterol. PMID- 16314225 TI - Cardiovascular risk in the Mediterranean area is low: impact on clinical decision making. PMID- 16314226 TI - The relationship between fish consumption and the risk of developing acute coronary syndromes among smokers: the CARDIO2000 case-control study. AB - BACKGROUND AND AIM: We sought to investigate the relationship between fish consumption and the risk of developing non-fatal acute coronary syndromes (ACS) among current smokers. METHODS AND RESULTS: During 2000-2001, we conducted the CARDIO2000 case-control study that included patients hospitalized for a first event of ACS and matched (age, sex) individuals without clinical evidence of CHD (controls), from all the Greek regions. In the present analysis we studied data from 519 men (59+/-10 years old) and 45 women (62+/-9 years old) patients, as well as 444 men and 90 women controls, matched for age with the patients that reported current smoking. To evaluate the hypothesis tested we applied conditional logistic regression analysis after controlling for several potential confounders. Overall, 94% of controls and patients reported fish consumption at least once a week. Compared to low fish consumption (<60 g/week), moderate (60 180 g/week) intake was associated with 15% (p<0.05) lower risk of ACS, while increased (180-300 g/week, or >300 g/week) consumption was associated with 18% (p<0.05) and 19% (p = 0.53) higher risk, respectively. CONCLUSION: Moderate fish consumption seems to moderate the deleterious effect of smoking on the occurrence of ACS. However, increased intake does not seem to confer any benefit on coronary risk. PMID- 16314227 TI - Metabolic and endocrine effects of physiological increments in plasma ghrelin concentrations. AB - BACKGROUND: Growing evidence indicates that the administration of large amounts of ghrelin to humans increases circulating concentrations of several pituitary and adrenal hormones, induces hyperglycemia and reduces serum insulin concentrations. At present, it is not known whether physiological increments in plasma ghrelin concentrations affect glucose kinetics or hormone concentrations in humans. METHODS AND RESULTS: We compared the effects of two- and three-fold increments in plasma ghrelin concentrations in eight healthy subjects during a 2 h intravenous infusion of 7.5 (GHRE7.5), 15 (GHRE15) pmol kg(-1) min(-1) acylated human ghrelin or placebo (PL), in a randomized double-blind study. Compared with PL (146 +/- 24 pM) plasma ghrelin concentrations increased at 120 min (p<0.001) about two-fold after GHRE7.5 (300 +/- 35 pM) and three-fold after GHRE15 (494 +/- 30 pM). GHRE15 significantly increased circulating concentrations of NEFA, GH, ACTH, epinephrine, and prolactin (p<0.01). GHRELIN7.5 significantly (p<0.01) increased only serum GH concentrations. Neither ghrelin infusions changed glucose flux or circulating concentrations of glucose, insulin, C-peptide, glucagon, IGF 1, cortisol and norepinephrine. CONCLUSIONS: GH secretion is the only response that is stimulated by physiological increments in plasma ghrelin concentrations; about three-fold increases in plasma ghrelin concentrations are required to elicit the responses of epinephrine, prolactin, ACTH and NEFA. PMID- 16314228 TI - Iron status and clinical outcome in patients with coronary artery disease after coronary stenting. AB - BACKGROUND AND AIM: So far, no studies have assessed whether there is an association between iron status and the incidence of major adverse cardiac events or restenosis after coronary stenting. We conducted this study to investigate whether there is an association between body iron status and clinical outcome in patients with coronary artery disease after coronary stenting. METHODS AND RESULTS: The study included 664 patients with coronary artery disease who underwent coronary stent implantation. The soluble transferring receptor/ferritin ratio (sTfR/ferritin ratio) was used as an index of iron status. Patients were divided into three groups according to the tertiles of sTfR/ferritin ratio: lower tertile (<11.9; n = 221), middle tertile (11.9-27.8; n = 221) and upper tertile (>27.8; n = 222). The combined incidence of major adverse cardiac events (death, myocardial infarction and target vessel revascularization) was the primary end point of the study. Patients in the lower tertile of the sTfR/ferritin ratio presented more often with unstable angina or acute myocardial infarction and had longer lesions and higher grade of stenosis than the patients in the middle or upper tertile of the sTfR/ferritin ratio. Angiographic restenosis at 6-month angiography was also evaluated. The cumulative event rate of composite end point of death, myocardial infarction or target vessel revascularization was 27.6% in patients in the lower tertile, 24.4% in patients in the middle tertile and 28.4% in patients in the upper tertile of the sTfR/ferritin ratio (p = 0.68). Restenosis was found in 27.8% (n = 45) in the lower tertile, 25.8% (n = 42) in the middle tertile and 27.5% (n = 38) in the upper tertile of the sTfR/ferritin ratio (p = 0.90). CONCLUSIONS: Our study showed no association between iron status and the incidence of major adverse cardiac events or angiographic coronary restenosis in patients with coronary artery disease after coronary stenting. PMID- 16314229 TI - Riskard 2005. New tools for prediction of cardiovascular disease risk derived from Italian population studies. AB - BACKGROUND AND AIM: The need to update tools for the estimate of cardiovascular risk prompted the "Gruppo di Ricerca per la Stima del Rischio Cardiovascolare in Italia" to produce a new chart and new software called Riskard 2005. METHODS AND RESULTS: Data from 9 population studies in 8 Italian regions, for a grand total of 17,153 subjects (12,045 men and 5,108 women) aged 35-74 and for a total exposure of about 194,000 person/years were available. A chart for the estimate of cardiovascular risk (major coronary, cerebrovascular and peripheral artery disease events) in 10 years was produced for men and women aged 45-74 free from cardiovascular diseases. Risk factors employed in the estimate were sex, age (6 classes), systolic blood pressure (4 classes), serum cholesterol (5 classes), diabetes, and cigarette smoking (4 classes). Estimates were produced for absolute risk and for relative risk, the latter against levels expected in the general population that produced the risk functions. Software was produced for the separate estimate of major coronary, cerebrovascular and cardiovascular events (the latter made by coronary, cerebrovascular and peripheral artery disease of atherosclerotic origin) for follow-up at 5, 10 or 15 years, in men a women aged 35-74 years at entry and free from cardiovascular diseases. Risk factors employed here were sex, age, body mass index, mean physiological blood pressure, HDL cholesterol, non-HDL cholesterol, cigarette smoking, diabetes and heart rate. The output is based on several indicators: absolute risk, relative risk (as defined above), ideal risk (for a very favourable risk profile), biological age of risk, comparisons among the above indicators, the percent contribution of risk factors to the excess of estimated risk above the level of the ideal risk, and the description of trends in risk estimate in relation to repeated measurements. CONCLUSIONS: These tools represent progress compared to similar tools produced some years ago by the same Research Group. PMID- 16314230 TI - Relationship between autonomic dysfunction, insulin resistance and hypertension, in diabetes. AB - Sympathovagal imbalance and insulin resistance are the common underlying disorders linking hypertension and diabetes. The role of hyperinsulinemia, however, on sympathovagal balance and blood pressure has never been clearly dissected from that of hyperglycemia. Nevertheless, the study of animal models of hypertension showed that hypertension does not invariably result in the onset of insulin resistance. This suggests that insulin resistance precedes the onset of hypertension and (possibly) contributes to its pathogenesis, mainly through sympathetic activation. To examine this hypothesis, recent studies investigated the relationship between insulin sensitivity and sympathetic activity in subjects with insulin resistance but free of overt hyperglycemia and obesity, i.e., insulin-resistant offspring of type 2 diabetic patients, demonstrating a prevalence of sympathetic over vagal activity. Therefore insulin resistance and sympathovagal imbalance come before hypertension, but a clear causative role cannot be demonstrated since other mechanisms, including an inappropriate lifestyle, must be taken into account to determine clinical hypertension. Finally, several experiments in human healthy volunteers suggest that the modulation of autonomic regulation at the forearm level can regulate insulin sensitivity, tempting us to speculate that it is the primary autonomic imbalance, through vasoconstriction, that results in both insulin resistance and hypertension. In conclusion, the close relationship between autonomic imbalance, insulin resistance and hypertension is unquestionable; although logical hypothesis can be constructed, which of the three is the earliest event is still not understood, and further research is required. PMID- 16314231 TI - Apolipoprotein CI, and not apolipoprotein E, polymorphism affects plasma levels of C-reactive protein? PMID- 16314232 TI - Elevated nitric oxide levels inhibit malondialdehyde production in patients with diabetes mellitus in Taiwan. PMID- 16314233 TI - Internal medicine training in Europe. PMID- 16314234 TI - Sweet's syndrome and sarcoidosis. AB - In this review we summarize a number of cases of Sweet's syndrome (SS) associated with sarcoidosis that have been reported in the English literature. In all of the cases, the two disorders were diagnosed simultaneously. Patients with both disorders were younger and had a higher rate of fever than patients with SS alone. In this group of patients, we found a trend toward less skin involvement of the face and trunk, more involvement of the upper limbs, and more atypical skin lesions, particularly papules. The association of the two disorders seems to be more related to a subset of acute sarcoidosis (Lofgren's syndrome). All of the patients in this group had a benign course and self-limiting disease. Thus, SS in association with sarcoidosis could be considered a favorable prognostic factor. Although SS has a high rate of recurrence, no recurrence occurred in this group of patients during follow-up. PMID- 16314235 TI - Disseminated intravascular coagulopathy in aortic aneurysms. AB - Aneurysm-induced disseminated intravascular coagulopathy (DIC) constitutes a rare presentation form of aortic aneurysms. The majority of DIC cases are asymptomatic and this condition is usually diagnosed during the perioperative workup; yet, in a minority of cases, DIC leads to the diagnosis of the vascular abnormality. The management of aneurysm-induced DIC is based both on the treatment of the underlying disorder and on an active scrutiny of the hemodynamic and blood support requirements. Blood replacement therapy should be individualized, guided by the clinical situation of the patient (especially considering the bleeding risk or the presence of hemorrhages), and accompanied by a close monitoring of the coagulation status. Fresh frozen plasma is usually the preferred initial option to replace coagulation factors, but fibrinogen, cryoprecipitates, and platelet concentrates are adequate options in certain contexts. Heparins, both non-fractionated and low-weight molecular types, are the most widely accepted agents for achieving adequate control of the coagulation activation and consumption. Other antithrombotic drugs are under study, including antithrombin III and activated protein C, although only the latter has demonstrated a benefit in terms of survival in a comparative, randomized context. Antifibrinolytic agents such as gabexate mesilate, tranexamic acid, and epsilon-aminocaproic acid (EACA) have been used with conflicting results. These agents may have a role for patients with catastrophic bleeding resistant to other therapeutical options, but their relevance as a first line of treatment is, at present, undefined. An assessment of the multitude of therapeutic approaches available would seem to indicate that there is a lack of standardization in the management of these patients. Multi-center, randomized clinical trials are needed to elucidate the most adequate therapy in this context. PMID- 16314236 TI - Postmenopausal women with Colles' fracture have bone mineral density values similar to those of controls when measured with calcaneus quantitative ultrasound. AB - BACKGROUND: It is a matter of controversy whether or not Colles' fracture is an osteoporotic fracture. Indeed, the usefulness of quantitative ultrasound in distinguishing Colles' fracture from normal fractures is also unclear. METHODS: A cross-sectional case-control study was done on 469 postmenopausal Spanish women, 121 with Colles' fracture and 348 controls. Assessment of risk factors for osteoporosis and measurement of calcaneus quantitative ultrasound were carried out using a Sahara, Hologic device. RESULTS: Patients with Colles' fracture had BUA, SOS, and QUI values that were similar to those of controls, and no statistically significant differences were found. We estimated ROC curves for SOS and a score based on a linear combination of height and SOS (SH-Score). The areas under both curves were 0.56 and 0.61, respectively, which was statistically significant. To obtain 5% false-negative and 10% false-positive figures, the T score cut-off for SOS was -2.45 and -0.045, respectively. Of these, only 9.2% were classified as high risk and 11% as low risk with 79.8% undetermined. CONCLUSIONS: Patients with Colles' fracture had BUA, SOS, and QUI values that were similar to those of controls. Nevertheless, ROC curves calculated by a combination of height and SOS showed that quantitative calcaneus ultrasound may be a useful tool for identifying postmenopausal women with Colles' fracture. These results indicate that measuring bone mineral density with ultrasound only captures limited aspects of the pathophysiology of Colles' fractures. PMID- 16314237 TI - Bacterial etiology of diabetic foot infections in South India. AB - BACKGROUND: Foot infections are a frequent complication of patients with diabetes mellitus, accounting for up to 20% of diabetes-related hospital admissions. Infectious agents are associated with the worst outcomes, which may ultimately lead to amputation of the infected foot unless prompt treatment strategies are ensued. The present study sought to reveal the bacterial etiology of diabetic foot ulcers in South India, the diabetic capital of India. METHODS: A 10-month long descriptive study was carried out to analyse the aerobic and anaerobic bacterial isolates of all patients admitted with diabetic foot infections presenting with Wagner grade 2-5 ulcers. Bacteriological diagnosis and antibiotic sensitivity profiles were carried out and analysed using standard procedures. RESULTS: Diabetic polyneuropathy was found to be common (56.8%) and gram-negative bacteria (57.6%) were isolated more often than gram-positive ones (42.3%) in the patients screened. The most frequent bacterial isolates were Pseudomonas aeruginosa, Staphylococcus aureus, coagulase-negative staphylococci (CONS), and Enterobacteriaceaes. Forty-nine cultures (68%) showed polymicrobial involvement. About 44% of P. aeruginosa were multi-drug-resistant, and MRSA was recovered on eight occasions (10.3%). Bacteroides spp. and Peptostreptococcus spp. were the major anaerobic isolates. CONCLUSIONS: Our study supports the viewpoint put forth by previous South Indian authors that the distribution of gram-negative bacteria (57.6%) is more common than that of gram-positive ones (42.3%) and it is contrary to the viewpoint that diabetic foot infections are frequently monomicrobial. Furthermore, recovery of multi-drug-resistant P. aeruginosa isolates is of serious concern, as almost no one has reported the same from the South Indian milieu. PMID- 16314238 TI - Safety and efficacy of percutaneous CT-guided liver biopsy using an 18-gauge automated needle. AB - BACKGROUND: Computed tomography (CT)-guided liver biopsy using large-caliber cutting needles allows the recovery of larger tissue specimens that are more suitable for establishing histological diagnosis. Yet, large-caliber needles are associated with higher rates of post-biopsy bleeding. The aim of this prospective study was to assess the efficacy and safety of the method, when all of the procedures were performed by experienced radiologists and all patients were carefully evaluated and treated, when needed, before the biopsy. METHODS: A total of 767 consecutive patients with focal hepatic lesions underwent CT-guided liver biopsy during a 5-year period. The procedures were performed in a single center using 18-gauge automated biopsy guns with a 2-cm cutting edge by a team of experienced radiologists (>100 procedures performed by each one before the initiation of the study). Before the procedure, abnormal coagulation indices were corrected, ascites was treated and, in all cases, an adequate parenchymal cuff of normal tissue between the lesion and the capsule was retained. RESULTS: In all cases the extracted specimens were adequate for diagnosis. No major complications (i.e., death or complications requiring surgery, chest tube, or blood transfusions) were observed. Minor complications (i.e., those not requiring medical intervention) were observed in three patients. CONCLUSIONS: Percutaneous CT-guided liver biopsy using an 18-gauge automated needle is a safe and effective procedure. Careful pre-biopsy evaluation and treatment, when needed, and maintenance of an adequate parenchymal cuff between the lesion and the capsule contribute to the safety of the method. PMID- 16314239 TI - Systematic diagnostic approach to proximal-without-distal claudication in a vascular population. AB - BACKGROUND: Very few observations of proximal-without-distal claudication have been reported in the literature. This is likely due to the use of questionnaires limiting vascular claudication to the calves and to the problems encountered in attributing unexplained "buttock" claudication to a vascular origin. METHODS: During a 2 1/2-year period, we searched for proximal-without-distal exercise related pain with the San Diego claudication questionnaire among some 2000 patients referred for lower limb arterial investigations. Of these patients, 97 presented no contraindication to treadmill testing and were investigated with exercise transcutaneous oxygen pressure (tcpO2). We used buttock tcpO2 (DROP index<-15 mm hg) to argue for the presence of ischemia on the corresponding side. RESULTS: Ischemia consistent with symptoms was found in 61 patients, whereas pain on one or both sides without underlying ischemia was found in 36 patients, suggesting a non-arterial origin of the symptoms. More than half of the patients with proximal-without-distal claudication and underlying exercise-related ischemia had been suffering for more than 2 years before they were referred to the laboratory. Eleven of the patients were treated. The treatment was successful in all but one of them. CONCLUSIONS: An important delay before diagnosis is frequently observed in proximal-without-distal claudication. TcpO2 is useful in attributing proximal exercise-related pain to a vascular origin. Given the number of detected and successfully treated patients in this small monocentric study, it is surprising that so few observations have been published to date, suggesting that proximal-without-distal arterial claudication is most likely an underestimated diagnosis. PMID- 16314240 TI - Arterial stiffness index as a screening test for cardiovascular risk: a comparative study between coronary artery calcification determined by electron beam tomography and arterial stiffness index determined by a VitalVision device in asymptomatic subjects. AB - BACKGROUND: Arterial stiffness has recently been proposed as a powerful independent predictor of cardiovascular disease. However, the influence of arterial stiffening on the interaction between the heart and large vessels and atherosclerosis is not well defined. The arterial stiffness index (ASI) has recently been determined with a new device (VitalVision) that calculates ASI in the upper arm using computerized oscillometry. Coronary artery calcification (CAC) is a useful surrogate marker of coronary artery disease detected non invasively by electron beam tomography (EBT). We investigated the correlation between ASI and CAC in a group of patients. METHODS: CAC and ASI measurements were determined with EBT and a VitalVision device, respectively, on the same day in 97 asymptomatic patients. Patients with calcium scores above 0 were classified as CAC+ and those with calcium scores equal to 0 were classified as CAC-. The ASI index was divided into three groups - mild, moderate, and high - according to the H-value, provided by the VitalVision device. RESULTS: In patients below 51 years of age, no correlation between the ASI and CAC was found. In patients over 50 years of age, a moderate positive and significant correlation was found between the CAC score and ASI measurements (r=0.40, p=0.001). CONCLUSIONS: The presence of a correlation between the CAC and ASI in patients over 50 shows that the ASI can be used to investigate atherosclerotic risk. PMID- 16314241 TI - Deliberate self-poisoning with acetaminophen: a comparison with other medications. AB - BACKGROUND: Acetaminophen is the most common drug employed in deliberate self poisoning (DSP) in many countries and can lead to acute liver failure. The purpose of the present study was to compare DSP with acetaminophen with DSP with other medications. METHODS: From January 2001 to December 2002, all patients admitted after DSP to Soroka University Hospital in Beer-Sheva, Israel, were identified. Demographic data and medical history were obtained retrospectively. RESULTS: There were 282 incidences of DSP in the study period. The mean age of 94 patients with an acetaminophen overdose was lower than that of patients who had attempted DSP with other medications (24.8 vs. 34.6 years, p<0.001). There was no gender or ethnic difference between the groups. Patients with DSP with acetaminophen were less likely to make recurrent attempts (23.4% vs. 39.9% p=0.006), less likely to have psychiatric disorders (32% vs. 59% p<0.001), and had a shorter hospital stay (1.7 vs. 2.1 days, p=0.05). The acetaminophen group had less of an impairment in level of consciousness (84% vs. 52.1% p<0.001) and had less need for ICU care (6.4% vs. 14.9%, p=0.04). Serum levels were checked in 68 (72.3%) of the patients in the acetaminophen group and they were found to be high in 6 (8.8%) of them (95% CI 2.1-15.5%). CONCLUSION: Taking an overdose of acetaminophen is a common method of DSP, with patients tending to have a more benign presentation and shorter hospital stay than those who attempt DSP with other medications. Further research on the optimal care of these patients and multi-disciplinary approaches to DSP prevention are needed. PMID- 16314242 TI - Internal medicine residency training in Turkey. AB - Medical school entrance depends on passing a central examination that is given annually by the National Selection and Placement Center. Undergraduate medical education takes 6 years. About 5000 students graduate from medical faculties annually. The central exam necessary for residency training is given by the National Selection and Placement Center. A Specialist Training Regulation regulates residency training. Internal medicine residency training takes 4 years and includes inpatient and outpatient care in wards and rotations. Residents prepare a dissertation that is used in the evaluation of residency competency. At the end of the residency period, residents who have been successful in previous evaluations take an oral exam followed by a written exam, which lead to their certification in internal medicine. Residents' scientific knowledge and skills are assessed by a jury consisting of five people, four from the same department and one from the equivalent department in another training institution. The title of specialist is granted after a certification exam given by training institutions and approved by the Ministry of Health. Internists are mainly employed in state hospitals, which are under the Ministry of Health. Subspecialty areas in internal medicine include gastroenterology, geriatrics, endocrinology, nephrology, hematology, rheumatology, immunology, allergology, and oncology. The training period for a subspecialty is 2 years. A substantial effort is being made all over the country to improve regulations and health care service delivery. These changes will also affect the residency training and manpower planning and employment of internists. PMID- 16314243 TI - Extramedullary hematopoiesis with spinal cord compression in beta-thalassemia intermedia. AB - Spinal cord compression due to extramedullary hematopoiesis is a rare manifestation of thalassemia. We present a 28-year-old woman with beta thalassemia intermedia and progressive paraparesis. She had a thoracic extradural extramedullary mass lesion on MRI. She improved after receiving multiple transfusions. Clinical awareness of this phenomenon with early treatment is essential for a successful outcome. PMID- 16314244 TI - Latent acute promyelocytic leukemia t(15;17)(q22;q12-21) and sarcoidosis: long term cohabitation. AB - The association of sarcoidosis with hematological malignancies is a well-known phenomenon. To our knowledge, we report the first case involving sarcoidosis and acute promyelocytic leukemia (APL) t(15;17)(q22;q12-21). The major interest lies in the chronology of the two diseases: the APL demonstrated an unusual smoldering evolution, suggesting that pre-existing sarcoidosis may have a non-fortuitous immunological impact on leukemic clone proliferation. PMID- 16314245 TI - Pulmonary and myocardial manifestations due to Toxocara canis infection. AB - We report a case of Toxocara canis infection manifested by persistent eosinophilia, pulmonary nodules on CT scan, and myocardial manifestations in a 38 year-old man who was a dog trainer. Toxocariasis is not common in adults and rarely affects the heart. PMID- 16314246 TI - Extensive myofascial necrosis: a delayed complication of botulinum toxin therapy. AB - Botulinum toxin is now an important therapeutic agent in various clinical specialties. Although a lot is known about its short-term effects, little is known about the long-term effects or delayed complications. Most of the therapeutic effects appear within a week and last for 10-12 weeks. In most cases, the side effects are mild and often self-limiting and tend to occur within the first week. Myofascial necrosis and delayed onset of side effects are rarely reported and present a new challenge for a drug that has been proclaimed as an anti-ageing drug by the lay press and is widely used outside the licensed indications. PMID- 16314247 TI - A mediastinal mass: Graves' disease related thymic hyperplasia. AB - A 35-year-old woman presented with symptoms of pulmonary embolism and thyrotoxicosis. Computed tomography of the chest ruled out pulmonary embolism but showed an incidental mediastinal mass. Graves' disease was confirmed and the mediastinal mass was ascribed to associating thymic hyperplasia. Treatment of Graves' thyrotoxicosis resulted in a spontaneous shrinkage of the mediastinal mass. Although it is relatively unknown, thymic hyperplasia is a normal and reversible manifestation of Graves' disease. PMID- 16314248 TI - Polysplenia in an elderly male: diagnostic approaches and review of the literature. AB - Polysplenia is a congenital disorder affecting the asymmetric organs, including the heart, lungs, bronchi, liver, stomach, pancreas, intestines, and spleen. Primary manifestations are heterogeneous and can include congenital heart disease, atypical placement of major abdominal organs, biliary atresia, and intestinal malrotation. Knowledge of these congenital abnormalities will enhance patient care. In this report, we present an elderly, asymptomatic individual with polysplenia to highlight the cardinal features and management issues. PMID- 16314249 TI - Cat scratch disease encephalopathy in an immunocompetent patient. AB - Cat scratch disease (CSD) is typically a self-limited regional lymphadenopathy in children and young adults that is caused by Bartonella henselae. The majority of CSD cases resolve spontaneously; however, many systemic complications have been described. We report an unusual case of CSD presenting as an epitrochlear arm mass and complicated by encephalopathy. Identification of B. henselae DNA in the affected lymph node and cerebrospinal fluid confirmed the diagnosis of CSD. Systemic antibiotic therapy was administered and the patient improved without any neurological deficit. PMID- 16314250 TI - Systemic allergic reaction and diffuse bone pain after exposure to a preparation of betamethasone. AB - Allergic reactions to corticosteroids are unexpected as they seem to contradict their pharmacodynamic action. Nevertheless, they are not infrequent, with an estimated incidence of up to 4% for cutaneous reactions. Systemic reactions are rarely reported, but their incidence might be underestimated. We report here an unusual allergic reaction to betamethasone presenting with diffuse bone pain, erythema, and bronchoconstriction, which was confirmed by a positive rechallenge in a double-blind procedure. This is the first case report of a systemic reaction to betamethasone confirmed by a positive rechallenge. An impurity in betamethasone diproprionate cannot be excluded. As this substance is frequently used in rheumatologic soft-tissue injections, it is important to recognize this potentially life-threatening side effect. PMID- 16314251 TI - Lipoid pneumonia: a preventable form of drug-induced lung injury. AB - We report a case where recurrent "pneumonia" was eventually diagnosed as lipoid pneumonia in an elderly patient with cerebrovascular disease. The discontinuation of paraffin oil laxative led to clinical improvement. Lipoid pneumonia, a foreign body-type reaction to the presence of lipid within lung parenchyma, is probably underdiagnosed and underreported, and paraffin oil laxative is the main causative agent. Paraffin oil is marketed as a food additive, and no information about its hazards is provided to clinicians or patients. We suggest that a change in paraffin oil licensing may decrease the incidence of lipoid pneumonia. PMID- 16314252 TI - Infective tricuspid valve endocarditis following fertility treatments. PMID- 16314253 TI - Isolated lymph node amyloidosis. PMID- 16314254 TI - Digital necrosis related to intralesional bleomycin. PMID- 16314255 TI - Asthma as well as anxiety improved by the Kampo extract Saiboku-to. PMID- 16314256 TI - Multiple lytic lesions on skull radiography: a diagnostic impasse. PMID- 16314257 TI - Methods for the study of alternative splicing. AB - Alternative splicing multiplies the coding capacity of genes, which provides unparalleled complexity to the transcriptome and proteome. This increased complexity has clear repercussions for the regulation of gene expression in many organisms and for the balance between human health and disease. This significance, which has only recently been fully understood, has led to a growing number of studies that address the mechanisms that control alternative splicing decisions and the biological consequences of alternative RNA and protein isoforms. These studies have adapted or developed new methods and protocols that span computational biology, the biochemical analysis of in vitro reactions, model tissue culture systems, and in vivo studies. Many of these methods have been assembled in this volume to assist a newcomer to the study of alternative splicing and to expand the methodologies of laboratories already in the field. PMID- 16314258 TI - Computational searches for splicing signals. AB - The removal of introns from pre-mRNA requires as an initial event the accurate molecular recognition of the proper exon-intron borders. It is now evident that RNA sequence elements in addition to the consensus splice site sequences themselves are required for this recognition. Genomic analyses have contributed to the definition of these elements as exonic and intronic splicing enhancers and silencers, comprising what has been called the "splicing code." Many computational methods have been brought to bear in such studies. We describe here some of the methods we have used to discover functional splicing signals. What these methods have in common is a comparison of sequences in and around exons to sequences found elsewhere in the genome. We have especially made use of comparisons to "pseudo exons," intronic sequences resembling exons by virtue of being bounded by sequences indistinguishable from splice sites. Two computational strategies are emphasized: (1) the use of a machine learning technique in which a computational algorithm, a support vector machine, is first trained on known examples and then used to predict sequences associated with splicing; and (2) straight statistical analysis of differences between regions associated with exons and other regions in the genome. In most cases, the predictions made using these methods have been validated by subsequent empirical tests. An attempt has been made to make this description understandable by researchers unfamiliar with computational practice and to include practical references to specific databases and programs. PMID- 16314259 TI - Analyzing mechanisms of alternative pre-mRNA splicing using in vitro splicing assays. AB - The development of in vitro assays to analyze pre-mRNA splicing resulted in the discovery of many fundamental features characterizing splicing signals and the machinery that completes this process. Because in vitro assays can be manipulated by various biochemical approaches, the versatility of investigating alternative pre-mRNA splicing in the test tube appears endless. Importantly, modifications in reaction conditions can lead to the accumulation, isolation, and characterization of reaction intermediates, a prerequisite for gaining mechanistic insights into how the spliceosome carries out intron removal, and how regulatory elements assist the general splicing machinery in defining splice sites and alternative exons. These considerable experimental advantages have made the in vitro splicing system a standard assay, even though this approach is independent from RNA transcription and other RNA processing events, and in some respects deviates from the natural process of mRNA biogenesis. Here, we describe the tools and techniques necessary to carry out in vitro splicing assays. Analyses of various experimental designs are presented to highlight the approaches taken to gain insights into the mechanisms by which splice site recognition and activation are communicated with the general splicing machinery. Methods to measure the kinetics of splicing, to observe the formation of the pre-spliceosomal complexes, and to manipulate and modify the in vitro system to resolve the regulatory influences in alternative splicing are presented. PMID- 16314260 TI - In vitro coupled transcription splicing. AB - Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here, we describe an in vitro system capable of transcribing and splicing complex transcripts with three and four exons. We also demonstrate how the system can be used to study exon silencing in vitro. We believe that this system will be a useful tool to study the mechanisms that mediate the coupling of transcription and pre-mRNA processing. PMID- 16314261 TI - Splicing-active nuclear extracts from rat brain. AB - In the nervous system, alternative pre-mRNA splicing generates the diverse protein machineries needed for cell excitation and synaptic communication. Yet, many questions remain about how these mechanisms are regulated by RNA binding proteins in the environment of differentiated cells and tissues. Here, we describe the preparation and use of splicing active nuclear extracts derived from the cerebellum and cerebral cortex regions of rat brain as a resource for in vitro studies. These tissue-specific extracts promote the neuron-specific pathway of splicing, and display characteristic changes in hnRNP protein function and expression. These extracts can be used in combination with affinity selection and depletion/complementation assays to identify regulatory factors and to characterize their interactions and effects on spliceosome assembly. The ability to prepare extracts from brain regions at a range of postnatal ages provides opportunities to address related questions as a function of cell differentiation. These neuronal extracts may also be valuable for the development of in vitro assays to elucidate other neuron-specific RNA processing pathways, such as 3' end formation, RNA editing, or miRNA maturation. PMID- 16314262 TI - Use of minigene systems to dissect alternative splicing elements. AB - Pre-mRNA splicing is an essential step for gene expression in higher eukaryotes. The splicing efficiency of individual exons is determined by multiple features involving gene architecture, a variety of cis-acting elements within the exons and flanking introns, and interactions with components of the basal splicing machinery (called the spliceosome) and auxiliary regulatory factors which transiently co-assemble with the spliceosome. Both alternative and constitutive exons are recognized by multiple weak protein:RNA interactions and different exons differ in the interactions which are determinative for exon usage. Alternative exons are often regulated according to cell-specific patterns and regulation is mediated by specific sets of cis-acting elements and trans-acting factors. Transient expression of minigenes is a commonly used in vivo assay to identify the intrinsic features of a gene that control exon usage, identify specific cis-acting elements that control usage of constitutive and alternative exons, identify cis-acting elements that control cell-specific usage of alternative exons, and once regulatory elements have been identified, to identify the trans-acting factors that bind to these elements and modulate splicing. This chapter describes approaches and strategies for using minigenes to define the cis acting elements that determine splice site usage and to identify and characterize the trans-acting factors that bind to these elements and regulate alternative splicing. PMID- 16314263 TI - Use of RNA interference to dissect the roles of trans-acting factors in alternative pre-mRNA splicing. AB - RNA interference (RNAi) is becoming a popular method for analyzing gene function in a variety of biological processes. We have used RNAi in cultured Drosophila cells to identify trans-acting factors that regulate the alternative splicing of endogenously transcribed pre-mRNAs. We have generated a dsRNA library comprising approximately 70% of the Drosophila genes encoding RNA binding proteins and assessed the function of each protein in the regulation of alternative splicing. This approach not only identifies trans-acting factors regulating specific alternative splicing events, but also can provide insight into the alternative splicing regulatory networks of Drosophila. Here, we describe this RNAi approach to identify alternative splicing regulatory proteins in detail. PMID- 16314264 TI - Detection and measurement of alternative splicing using splicing-sensitive microarrays. AB - Splicing and alternative splicing are major processes in the interpretation and expression of genetic information for metazoan organisms. The study of splicing is moving from focused attention on the regulatory mechanisms of a selected set of paradigmatic alternative splicing events to questions of global integration of splicing regulation with genome and cell function. For this reason, parallel methods for detecting and measuring alternative splicing are necessary. We have adapted the splicing-sensitive oligonucleotide microarrays used to estimate splicing efficiency in yeast to the study of alternative splicing in vertebrate cells and tissues. We use gene models incorporating knowledge about splicing to design oligonucleotides specific for discriminating alternatively spliced mRNAs from each other. Here we present the main strategies for design, application, and analysis of spotted oligonucleotide arrays for detection and measurement of alternative splicing. We demonstrate these strategies using a two-intron yeast gene that has been altered to produce different amounts of alternatively spliced RNAs, as well as by profiling alternative splicing in NCI 60 cancer cell lines. PMID- 16314265 TI - Visualization of alternative splicing in vivo. AB - The analysis of sequences required for alternative splicing of mRNA has predominantly been performed using cell culture systems. However, the phenotype of cultured cells is almost invariably different from that of cells in the intact animal. It is therefore possible that there are significant differences in the regulation of specific splicing reactions in vivo compared to in cell culture. Here, we describe methods for the visualization and analysis of alternative splicing in vivo using transgenic mice. These methods allow for the analysis of the temporal and tissue-specific regulation of alternative splicing both visually and quantitatively. PMID- 16314266 TI - Stabilization and analysis of intron lariats in vivo. AB - The analysis of lariats produced in vivo during pre-mRNA splicing is a powerful tool for elucidation of regulatory mechanisms and identification of natural recursive splicing events. Nevertheless, this analysis is technically challenging because lariats normally have short half-lives. With appropriate controls, RT-PCR amplification and sequencing of the region spanning the 2'-5' phosophodiester bond at the branch junction can be a sensitive and versatile method for lariat analysis. This approach can be facilitated and enhanced by reducing the activity of debranching enzyme (DBR) in order to stabilize lariats. We have generated a set of plasmids for dsRNA-mediated knockdown of DBR under diverse conditions in transgenic Drosophila and in cultured cells. We describe the use of these plasmids and protocols for lariat analysis. We have generated transgenic Drosophila strains carrying a GAL4-regulated RNAi construct that allows selective knockdown of DBR in specific tissues or developmental stages, using the large collection of available GAL4 expression lines. These strains should prove useful for detailed developmental analyses of alternative and recursive splicing and for genetic analyses of splicing factors. Similar approaches should be readily adaptable to other organisms. PMID- 16314267 TI - CLIP: a method for identifying protein-RNA interaction sites in living cells. AB - Nucleic-acid binding proteins constitute nearly one-fourth of all functionally annotated human genes. Genome-wide analysis of protein-nucleic acid contacts has not yet been performed for most of these proteins, restricting attempts to establish a comprehensive understanding of protein function. UV cross-linking is a method typically used to determine the position of direct interactions between proteins and nucleic acids. We have developed the cross-linking and immunoprecipitation assay, which exploits the covalent protein-nucleic acid cross linking to stringently purify a specific protein-RNA complex using immunoprecipitation followed by SDS-PAGE separation. In this way, the vast majority of non-specific contaminating RNA, which can bind to co immunoprecipitated proteins or beads, can be removed. Here, we present an improved protocol that performs RNA linker ligation before the SDS-PAGE step, and describe its application to the specific purification and amplification of RNA ligands of Nova in neurons. PMID- 16314268 TI - Conditional knockout mice to study alternative splicing in vivo. AB - Analysis of genomes has revealed that the total number of human genes is comparable to those of simpler organisms, and thus, the number of genes does not correlate with the complexity and functional diversity of different organisms. Multiple mechanisms, including alternative splicing, are believed to contribute to the molecular complexity in higher eukaryotes. Given the fact that more than half of human genes undergo alternative splicing, however, little is known about the biological relevance of most alternative splicing events and their regulatory mechanisms. Recent work has highlighted the power of reverse genetic approaches in addressing regulated splicing in animal models. Here, we focus on the conditional knockout approach adapted for splicing research with the intention to provide a general guide to the generation of mouse models to study regulated splicing in development and disease. PMID- 16314270 TI - Hemolytic-uremic syndrome: illustrative cases and review. PMID- 16314275 TI - Life Flight celebrates silver anniversary in Jacksonville. PMID- 16314276 TI - The roadside high: Jimson weed toxicity. PMID- 16314277 TI - Helicopter emergency medical service in Italy. PMID- 16314278 TI - Effect of transport team interventions on stabilization time in neonatal and pediatric interfacility transports. AB - INTRODUCTION: During interfacility transport, the length of time taken by the transport team to prepare the patient for transport is often perceived as a problem by referring hospital staff. The purpose of this study was to examine the effects on time at the referring hospital of the number and complexity of interventions performed by the transport team to stabilize the patient prior to transfer. SETTING: Interfacility transfers by the provincial infant transport team (ITT) to British Columbia's Children's Hospital. METHODS: This was a prospective study of emergency neonatal and pediatric interfacility transfers. After each transport, the team completed a questionnaire about interventions performed and stabilization time. Transports were classified by the complexity of interventions performed: none, low (intravenous line, blood gas, nasogastric tube, Foley catheter, oxygen administration), or high (intubation, central venous access, arterial lines, chest tube insertion). RESULTS: Thirty of 55 transports required no intervention (mean stabilization time=52+/-25 min). Sixteen transports required low level intervention (mean=60+/-22 min). Nine transports required high level intervention (mean=140+/-52 min). The stabilization times for "no" and "low" levels of intervention were not significantly different (P=.3), but the time for "high" level intervention was significantly higher (P<.01). CONCLUSIONS: The need for the transport team paramedics to perform high level interventions significantly increased the time at the referring hospital. In contrast, the time taken for them to perform or reperform low level interventions, whether one procedure or two, was not a significant source of delay. PMID- 16314279 TI - Physician-staffed HEMS dispatch in the Netherlands: Adequate deployment or minimal utilization? AB - INTRODUCTION: In the Netherlands, a physician-staffed helicopter emergency medical system (HEMS), called the Helicopter Mobile Medical Team (HMMT), provides prehospital care for severely injured patients in addition to ambulance services. This HMMT has proven to increase chances of survival and reduce morbidity. HMMT dispatch is performed following certain dispatch criteria. The goal of this study was to analyze actual dispatch rates and assess the protocol adherence of the emergency dispatchers in Rotterdam regarding HMMT dispatch. METHODS: All high priority ambulance runs between April 1 and July 1, 2003, were prospectively documented and cross-referenced to dispatch criteria. It was determined whether the emergency call warranted either immediate dispatch of the HMMT or a secondary dispatch after arrival of the first ambulance. When dispatch actually occurred, this was also documented. RESULTS: In The Studied Period A Total Of 5765 A1 Ambulance Runs During Daylight Were Documented. Of These, 1148 Runs Met Primary Dispatch Criteria And 38 Runs Met Secondary Dispatch Criteria. Actual Hmmt Dispatch Occured In 162/1186 (14%) Cases. CONCLUSIONS: HEMS dispatch rates and dispatch criteria adherence are low (14%). Better protocol adherence by emergency dispatchers could lead to a sevenfold increase of HMMT dispatches. The reasons for suboptimal protocol adherence remain unclear and persist, despite proven value of the HMMT in reducing patient mortality and morbidity. PMID- 16314280 TI - Occupational injuries and stressors among Canadian air medical health care professionals in rotor-wing programs. AB - INTRODUCTION: Air medical health care providers work in a unique environment that may affect occupational injury rates and patterns. Despite this knowledge, little high-quality evidence exists regarding occupational injuries specifically incurred by air medical health care professionals. We sought to characterize the epidemiology of occupational injuries experienced by Canadian rotor-wing health care providers. METHODS: A survey was sent to the 4 rotor-wing programs in Canada. All crewmembers participating directly in patient care were asked to complete the survey detailing any acute occupational injuries sustained within the previous year. A series of both open- and closed-ended questions was used to collect participant demographics and information regarding any injuries sustained. RESULTS: One hundred and six (40.6%) participants completed the survey. Three hundred and thirty acute injuries were reported. Hand lacerations and leg contusions were most prevalent (31 and 24 individuals incurred these injuries, respectively). Acute back injuries were also prevalent with 25 (23.6%) participants reporting at least one back injury. Overall, an injury rate of 3.2 injuries per person per year was reported. Lifting was cited as a common factor in injury (30 cases). Most injuries required little treatment, with only 17 needing physician intervention, and only 6 required more than 1 week off work. CONCLUSION: Injuries among Canadian air medical crews are common, but fortunately, the majority are minor. Specific injury prevention strategies may focus on stretcher design, cabin ergonomics, and extremity protective equipment. PMID- 16314281 TI - Monitoring the interaction between DNA and a transcription factor (MEF2A) using fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) is an analytical method that allows distinguishing different populations of fluorescent probes in solution and provides data on their concentrations and their diffusion coefficients. FCS was used to characterize the interaction of the transcription factor (MEF2A) with its DNA target sequence. The myocyte enhancer factor 2 (MEF2) belongs to the MADS-box family and activates transcription of numerous muscle genes during myogenesis. Measurements were made using TAMRA-labelled oligonucleotide duplexes derived from a wild type (WT) or a mutated MEF2 target gene. Binding of the protein to the WT DNA resulted in significant changes of the diffusion. Specificity of the interaction was confirmed using the mutated DNA. Bound to free probe ratios were determined at different MEF2A concentrations and the apparent equilibrium dissociation constant K(D) for the full-length MEF2A was estimated. PMID- 16314282 TI - Investigation by fluorescence correlation spectroscopy of the chaperoning interactions of HIV-1 nucleocapsid protein with the viral DNA initiation sequences. AB - HIV-1 nucleocapsid protein (NC) exhibits nucleic acid chaperone properties that are important during reverse transcription. Herein, we review and extend our recent investigation by fluorescence correlation spectroscopy (FCS) of the NC chaperone activity on the primer binding site sequences (PBS) of the (-) and (+) DNA strands, which are involved in the second strand transfer during reverse transcription. In the absence of NC, the PBS stem-loops exhibited a fraying limited to the terminal G-C base pair. The kinetics of fraying were significantly activated by NC, a feature that may favour (-)PBS/(+)PBS annealing during the second strand transfer. In addition, NC was found to promote the formation of PBS kissing homodimers through interaction between the loops. These kissing complexes may favour secondary contacts between viral sequences and thus, promote recombination and viral diversity. PMID- 16314283 TI - Disassembly of structurally modified viral nanoparticles: characterization by fluorescence correlation spectroscopy. AB - Analysis of the breakdown products of engineered viral particles can give useful information on the particle structure. We used various methods to breakdown both a recombinant enveloped virus and virus-like particles (VLPs) from two non enveloped viruses and analysed the resulting subunits by fluorescence correlation spectroscopy (FCS). Analysis of the enveloped baculovirus, Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), displaying the green fluorescent protein (GFP) fused to its envelope protein gp64 was performed in the presence and absence of 5 mM SDS and 25 mM DTT. Without treatment, the viral particle showed a diffusion time of 3.3 ms. In the presence of SDS, fluorescent subunits with diffusion times of 0.2 ms were observed. Additional treatment with DTT caused a drop in the diffusion time to 0.1 ms. Changes in the amplitude of the autocorrelation function suggested a 3-fold increase in fluorescent particle number when viral particles were treated with SDS, and a further 1.5-fold increase with additional treatment with DTT. Thus, the data showed that an average of 4.5 molecules of gp64-GFP was incorporated in the membrane of the modified baculovirus. Further, this suggests that each fluorescent gp64 trimer carries on average 1.5 fluorescent units. Similar experiments were carried out with two non-enveloped fluorescent virus-like particles (fVLPs) that displayed enhanced green fluorescent protein (EGFP). These, fVLPs of canine and human B19 parvoviruses were treated with 6 M urea and 5 mM SDS, respectively. Correspondingly, the original hydrodynamic radii of 17 and 14 nm were reduced to 9 and 5 nm after treatment. Here, the change in the amplitude of the autocorrelation curve suggested a 10-fold increase in particle number when viral particles of CPV were treated with 6 M urea at 50 degrees C for 10 min. For EGFP B19, there was a decrease in the amplitude, accompanied by a 9-fold increase in the number of fluorescent units with SDS treatment. The results showed that approximately 10 and 9 fluorescent units were associated with the corresponding CPV and B19 VLPs. In summary, we were able to estimate the number of fluorescent subunits in a baculovirus containing a GFP-fusion with its gp64 envelope protein and in two different parvo-VLPs containing EGFP-fused with their VP2 capsid proteins. PMID- 16314284 TI - Diffusion of sphingomyelin and myelin oligodendrocyte glycoprotein in the membrane of OLN-93 oligodendroglial cells studied by fluorescence correlation spectroscopy. AB - Evidence has been accumulated that the plasma membrane of various mammalian cell types is heterogeneous in structure and may contain lipid microdomains (lipid rafts). This study focuses on the membrane organization of living oligodendrocytes, which are the myelin-producing cells of the central nervous system. Fluorescence correlation spectroscopy (FCS) was used to monitor the lateral diffusion of a lipid and of a protein in the oligodendroglial cell line OLN-93. The lipid was fluorescently labelled sphingomyelin (Bodipy FL-C5 SM). The protein was the myelin oligodendrocyte glycoprotein (MOG). In order to monitor the lateral diffusion of MOG, OLN-93 cells were transfected with a MOG-EGFP (enhanced green fluorescent protein) fusion plasmid. The measurements were performed at room temperature. FCS data were analyzed for two-dimensional (2D) diffusion according to three models which all included a triplet fraction: (a) 2D 1 component (2D1C), (b) 2D anomalous diffusion (2D1Calpha), and (c) 2D 2 components (2D2C). Preliminary results indicate that for the lipid case, the best fits are obtained with 2D2C. In the case of MOG-EGFP, 2D2C and 2D1Calpha give fits of similar quality. The parameter estimates obtained with 2D1Calpha, however, have a lower standard deviation. The anomaly parameter for MOG-EGFP is 0.59+/-0.01. PMID- 16314285 TI - In situ measurements of viral particles diffusion inside mucoid biofilms. AB - Fluorescence correlation spectroscopy (FCS) under two-photon excitation was used successfully to characterize the diffusion properties of model virus particles (bacteriophages) in bacterial biofilm of Stenotrophonas maltophilia. The results are compared to those obtained with fluorescent latex beads used as a reference. The FCS data clearly demonstrated the possibility for viral particles to penetrate inside the exopolymeric matrix of mucoid biofilms, and hence to benefit from its protective effect toward antimicrobials (antibiotics and biocides). Microbial biofilms should hence be considered as potential reservoirs of pathogenic viruses, and are probably responsible for numerous persistent viral infections. PMID- 16314286 TI - Dissociation of nuclear import cargo complexes by the protein Ran: a fluorescence correlation spectroscopy study. AB - In nucleated cells, proteins designed for nuclear import form complexes with soluble nuclear transport receptors prior to translocation across the nuclear envelope. The directionality of transport is due to the asymmetric distribution of the protein Ran, which dissociates import cargo complexes only in its nuclear RanGTP form. Using fluorescence correlation spectroscopy, we have studied the stability of cargo complexes in solution in the presence and in the absence of RanGTP. We find that RanGTP has a higher affinity for the major import receptor, the importin alpha/beta heterodimer, when importin alpha does not carry a cargo, suggesting that some nuclear transport targets might be preferentially released. PMID- 16314287 TI - Controversies in the therapy of early stage breast cancer. AB - Breast cancer is the most common malignancy among U.S. women, with more than 200,000 new cases diagnosed annually. In the U.S., mortality from breast cancer has declined in recent years as a result of more widespread screening, leading to earlier detection, as well as advances in the adjuvant treatment of early-stage disease. It is widely accepted that the appropriate use of adjuvant chemotherapy and endocrine therapy improves the disease-free and overall survival of patients with early-stage breast cancer. It is, therefore, standard clinical practice to administer adjuvant systemic therapy to patients with node-positive and high risk, node-negative breast cancer. There remain, however, many controversies in the primary systemic therapy of breast cancer, which are discussed in this review. PMID- 16314288 TI - Adjuvant chemotherapy for early breast cancer: optimal use of epirubicin. AB - Anthracyclines are central components of adjuvant combination chemotherapy regimens for early breast cancer. Epirubicin is underutilized for this indication in the United States, where it was approved by the Food and Drug Administration in 1999, compared to Europe and Canada, where it gained approval in 1980. Use of epirubicin offers advantages in specific treatment settings and patient subsets, including situations where use of dose-dense and/or dose-intense protocols may provide additional benefits and where combinations including taxanes and/or trastuzumab may provide increased efficacy. Epirubicin also has a distinct safety profile compared to doxorubicin with regard to cardiotoxicity. In order to optimize treatment benefits and safety concerns for node-positive, node-negative and HER-2-positive patients as well as patients receiving neoadjuvant therapy and elderly patients it is worthwhile to consider the potential benefits of epirubicin. PMID- 16314289 TI - The role of taxanes in the management of bladder cancer. AB - Transitional cell carcinoma of the bladder is a chemo-sensitive neoplasm. Whereas the MVAC (methotrexate, vinblastine, doxorubicin, and cisplatin) regimen was long considered the standard of care for patients with advanced disease, the evaluation of newer agents with retained activity and improved tolerability has been the focus of much investigation over the past decade. Among the most important of these newer agents are taxanes. Whereas taxane-containing regimens have not yet been shown to improve the survival of patients with transitional cell carcinoma in randomized trials, ongoing phase III trials will further define the role of these agents in both the perioperative and advanced disease settings. PMID- 16314290 TI - When prostate brachytherapy fails: a case report and discussion. AB - For appropriately selected brachytherapy patients, prostate-specific antigen failure is uncommon. Our patient experienced biochemical failure after 125I brachytherapy treatment for low-risk prostate adenocarcinoma. We suggest neoadjuvant/adjuvant hormonal therapy combined with pelvic external-beam radiation therapy as a reasonable salvage treatment. At the 2-year follow-up, he is apparently doing well. With limited data available, salvage management for this situation is presently investigational. PMID- 16314291 TI - Practice points, consensus, and controversial issues in the management of patients with newly diagnosed acute promyelocytic leukemia. AB - Recent reviews on acute promyelocytic leukemia (APL) treatment have focused on comparing therapeutic approaches, including all-trans retinoic acid (ATRA) and chemotherapy, and do not address several other aspects of APL management that are relevant to the outcome in individual patients. These aspects include appropriate diagnostic tools and strategies, supportive care, recognition and treatment of life-threatening complications, evaluation of response, and, finally, management of the disease in special conditions such as older patients and pregnant women. In addition to reviewing current consensus and controversies of ATRA and chemotherapy treatment, this article addresses the above issues of APL management with special emphasis on aspects that distinguish APL from other acute myelogenous leukemias. PMID- 16314292 TI - Current therapy for Wilms' tumor. AB - Wilms' tumor was the first solid malignancy in which the value of adjuvant chemotherapy was established. Multimodality treatment has resulted in a significant improvement in outcome from approximately 30% in the 1930s to more than 85% in the modern era. Although the National Wilms' Tumor Study Group and the International Society of Pediatric Oncology differ philosophically regarding the merits of preoperative chemotherapy, outcomes of patients treated with either up-front nephrectomy or preoperative chemotherapy have been excellent. The goal of current clinical trials is to reduce therapy for children with low-risk tumors, thereby avoiding acute and long-term toxicities. At the same time, current clinical trials seek to augment therapy for patients with high-risk Wilms' tumor, including those with bilateral, anaplastic, and recurrent favorable histology tumors. PMID- 16314293 TI - ET-743: a novel agent with activity in soft tissue sarcomas. AB - Ecteinascidin-743 (ET-743) is a natural product derived from the marine tunicate Ectenascidia turbinate. ET-743 binds in the minor groove of DNA, blocks transcription factors activity, and traps protein from the nucleotide excision repair system, thus blocking cells in G2-M phase. ET-743 demonstrated cytotoxic activity at very low concentrations against sarcoma cell lines in pre-clinical studies. In several phase II clinical studies in patients with advanced sarcoma failing conventional doxorubicin- and ifosfamide-based chemotherapy, ET-743 delivered by continuous intravenous 24-hour infusion at a dose of 1,500 microg/m2 every 21 days yielded 8% overall response and 30%-40% stabilization rates for a clinical benefit rate close to 40%. Interestingly, long-term stabilizations over more than 3 years have been described. In vivo, ET-743 has a specific toxicity profile, the major toxicity of this product being hepatic, through biliary duct destruction, and hematologic. ET-743 has also been evaluated in first-line treatment for these patients. Finally, due to its original mode of action and the lack of cross-resistance with other chemotherapy agents, ET-743 was tested in a preclinical model in combination with other drugs. Synergy was reported in vitro with doxorubicin and cisplatin; phase I combination studies are in progress. PMID- 16314294 TI - Using single-agent therapy in adult patients with advanced soft tissue sarcoma can still be considered standard care. AB - The group of soft tissue sarcomas in adult patients is a heterogeneous group with more than 40 different subtypes. While local treatment remains the mainstay for localized disease, systemic chemotherapy can importantly contribute in the treatment of advanced soft tissue sarcoma. For patients with metastatic disease, chemotherapy is a palliative treatment in the vast majority of the cases. In this setting, toxicity should not outweigh the potential benefits resulting from chemotherapy. In patients with locally advanced disease too extensive for local treatment, systemic chemotherapy can contribute to cure, provided that tumor shrinkage renders subsequent optimal local treatment possible. In these cases, chemotherapeutic regimens yielding the highest response rates achievable should be used. In the last decades, several randomized studies have aimed to determine whether combination regimens yield benefit over single-agent treatment in terms of response rate and overall survival. This review addresses the current available data on chemotherapy for adult patients with soft tissue sarcoma, excluding gastrointestinal stromal tumor, the Ewing-like sarcomas, and other small blue round cell tumors. In addition, it is increasingly recognized that future research in soft tissue sarcoma should focus on the identification of tumor factors that can serve as targets for treatment and that the diverse tumor subtypes should be analyzed separately for their sensitivity to systemic treatment. This review also focuses on these and other strategies that will hopefully lead to better out comes in this disease entity in the near future. PMID- 16314295 TI - Renal safety and efficacy of i.v. bisphosphonates in patients with skeletal metastases treated for up to 10 Years. AB - INTRODUCTION: Bisphosphonates (BPs) delay the onset or reduce the incidence of skeletal complications in patients with bone metastases. However, there are few data on the renal safety and activity of i.v. BPs beyond 2 years of administration. MATERIALS AND METHODS: We retrospectively analyzed serum creatinine (SCr) levels and skeletal-related events (SREs) in cancer patients receiving i.v. BPs for >or= 24 months. All patients received 90 mg pamidronate every 3-4 weeks. Pre- and post-treatment SCr levels and the peak levels attained were recorded. A notable SCr increase was defined as: an increase >0.5 mg/dl for patients with baseline SCr <1.4 mg/dl; an increase >1 mg/dl for patients with baseline SCr >1.4 mg/dl; or doubling over baseline. The following parameters were also analyzed: the proportion of patients with at least one SRE, the distribution of each type of SRE, the time to first SRE, and the skeletal morbidity rate (SMR). RESULTS: Fifty-seven patients with bone metastases resulting from breast cancer (BC) (n = 48), multiple myeloma (n = 7), renal cell carcinoma (n = 1), and prostate cancer (n = 1) were evaluated. The median age at the start of treatment was 57 years (range, 27-81); 25% of the patients were >70 years old. Forty-three patients received pamidronate then switched to zoledronic acid. The median overall duration of BP administration was 34 months (range, 24+ to 131+), with a median duration of zoledronic acid therapy of 25 months (range, 2-40). Twenty seven of 48 BC patients received different chemotherapy regimens (median number of lines, 2; range, 1-6). The median SCr levels were: baseline, 0.82 mg/dl (range, 0.4-1.4); time of analysis, 0.89 mg/dl (0.4-2); highest level, 1.0 mg/dl (0.5-2). A notable SCr increase was observed in seven patients (12.2%; all grade 1). Twenty-six patients (45.6%) experienced SREs after starting BP treatment. The median time to first SRE was 911 days (95% confidence interval, 731; 1,023). The SMR was 0.20 events per year. Ten patients ceased treatment because of: an SCr level of 2 mg/dl (n = 1) physician decision (n = 6) and jaw osteonecrosis (n = 3). Ten patients died of progressive disease. CONCLUSION: i.v. BPs are safe and active during prolonged treatment administration, and renal function is maintained in patients receiving multiple cytotoxic therapies. Jaw osteonecrosis occurred in 5% of the study population, and its causal relationship with BP treatment requires further observation and study. PMID- 16314296 TI - Faster quantitative real-time PCR protocols may lose sensitivity and show increased variability. AB - Quantitative real-time PCR has become the method of choice for measuring mRNA transcription. Recently, fast PCR protocols have been developed as a means to increase assay throughput. Yet it is unclear whether more rapid cycling conditions preserve the original assay performance characteristics. We compared 16 primer sets directed against Epstein-Barr virus (EBV) mRNAs using universal and fast PCR cycling conditions. These primers are of clinical relevance, since they can be used to monitor viral oncogene and drug-resistance gene expression in transplant patients and EBV-associated cancers. While none of the primers failed under fast PCR conditions, the fast PCR protocols performed worse than universal cycling conditions. Fast PCR was associated with a loss of sensitivity as well as higher variability, but not with a loss of specificity or with a higher false positive rate. PMID- 16314297 TI - Allele quantification using molecular inversion probes (MIP). AB - Detection of genomic copy number changes has been an important research area, especially in cancer. Several high-throughput technologies have been developed to detect these changes. Features that are important for the utility of technologies assessing copy number changes include the ability to interrogate regions of interest at the desired density as well as the ability to differentiate the two homologs. In addition, assessing formaldehyde fixed and paraffin embedded (FFPE) samples allows the utilization of the vast majority of cancer samples. To address these points we demonstrate the use of molecular inversion probe (MIP) technology to the study of copy number. MIP is a high-throughput genotyping technology capable of interrogating >20 000 single nucleotide polymorphisms in the same tube. We have shown the ability of MIP at this multiplex level to provide copy number measurements while obtaining the allele information. In addition we have demonstrated a proof of principle for copy number analysis in FFPE samples. PMID- 16314298 TI - Enhancing the efficiency of a PCR using gold nanoparticles. AB - We found that the PCR could be dramatically enhanced by Au nanoparticles. With the addition of 0.7 nM of 13 nm Au nanoparticles into the PCR reagent, the PCR efficiency was increased. Especially when maintaining the same or higher amplification yields, the reaction time could be shortened, and the heating/cooling rates could be increased. The excellent heat transfer property of the nanoparticles should be the major factor in improving the PCR efficiency. Different PCR systems, DNA polymerases, DNA sizes and complex samples were compared in this study. Our results demonstrated that Au nanoparticles increase the sensitivity of PCR detection 5- to 10-fold in a slower PCR system (i.e. conventional PCR) and at least 10(4)-fold in a quicker PCR system (i.e. real-time PCR). After the PCR time was shortened by half, the 100 copies/microl DNA were detectable in real-time PCR with gold colloid added, however, at least 10(6) copies/microl of DNA were needed to reach a detectable signal level using the PCR reagent without gold colloid. This innovation could improve the PCR efficiency using non-expensive polymerases, and general PCR reagent. It is a new viewpoint in PCR, that nanoparticles can be used to enhance PCR efficiency and shorten reaction times. PMID- 16314300 TI - An obligate intermediate along the slow folding pathway of a group II intron ribozyme. AB - Most RNA molecules collapse rapidly and reach the native state through a pathway that contains numerous traps and unproductive intermediates. The D135 group II intron ribozyme is unusual in that it can fold slowly and directly to the native state, despite its large size and structural complexity. Here we use hydroxyl radical footprinting and native gel analysis to monitor the timescale of tertiary structure collapse and to detect the presence of obligate intermediates along the folding pathway of D135. We find that structural collapse and native folding of Domain 1 precede assembly of the entire ribozyme, indicating that D1 contains an on-pathway intermediate to folding of the D135 ribozyme. Subsequent docking of Domains 3 and 5, for which D1 provides a preorganized scaffold, appears to be very fast and independent of one another. In contrast to other RNAs, the D135 ribozyme undergoes slow tertiary collapse to a compacted state, with a rate constant that is also limited by the formation D1. These findings provide a new paradigm for RNA folding and they underscore the diversity of RNA biophysical behaviors. PMID- 16314301 TI - The genetic map and comparative analysis with the physical map of Trypanosoma brucei. AB - Trypanosoma brucei is the causative agent of African sleeping sickness in humans and contributes to the debilitating disease 'Nagana' in cattle. To date we know little about the genes that determine drug resistance, host specificity, pathogenesis and virulence in these parasites. The availability of the complete genome sequence and the ability of the parasite to undergo genetic exchange have allowed genetic investigations into this parasite and here we report the first genetic map of T.brucei for the genome reference stock TREU 927, comprising of 182 markers and 11 major linkage groups, that correspond to the 11 previously identified chromosomes. The genetic map provides 90% probability of a marker being 11 cM from any given locus. Its comparison to the available physical map has revealed the average physical size of a recombination unit to be 15.6 Kb/cM. The genetic map coupled with the genome sequence and the ability to undertake crosses presents a new approach to identifying genes relevant to the disease and its prevention in this important pathogen through forward genetic analysis and positional cloning. PMID- 16314302 TI - Simulation of non-specific protein-mRNA interactions. AB - Protein-nucleic acid interactions exhibit varying degrees of specificity. Relatively high affinity, sequence-specific interactions, can be studied with structure determination, but lower affinity, non-specific interactions are also of biological importance. We report simulations that predict the population of nucleic acid paths around protein surfaces, and give binding constant differences for changes in the protein scaffold. The method is applied to the non-specific component of interactions between eIF4Es and messenger RNAs that are bound tightly at the cap site. Adding a fragment of eIF4G to the system changes both the population of mRNA paths and the protein-mRNA binding affinity, suggesting a potential role for non-specific interactions in modulating translational properties. Generally, the free energy simulation technique could work in harness with characterized tethering points to extend analysis of nucleic acid conformation, and its modulation by protein scaffolds. PMID- 16314303 TI - Identification and characterization of high affinity antisense PNAs for the human unr (upstream of N-ras) mRNA which is uniquely overexpressed in MCF-7 breast cancer cells. AB - We have recently shown that an MCF-7 tumor can be imaged in a mouse by PET with 64Cu-labeled Peptide nucleic acids (PNAs) tethered to the permeation peptide Lys4 that recognize the uniquely overexpressed and very abundant upstream of N-ras or N-ras related gene (unr mRNA) expressed in these cells. Herein we describe how the high affinity antisense PNAs to the unr mRNA were identified and characterized. First, antisense binding sites on the unr mRNA were mapped by an reverse transcriptase random oligonucleotide library (RT-ROL) method that we have improved, and by a serial analysis of antisense binding sites (SAABS) method that we have developed which is similar to another recently described method. The relative binding affinities of oligodeoxynucleotides (ODNs) complementary to the antisense binding sites were then qualitatively ranked by a new Dynabead-based dot blot assay. Dissociation constants for a subset of the ODNs were determined by a new Dynabead-based solution assay and were found to be 300 pM for the best binders in 1 M salt. PNAs corresponding to the ODNs with the highest affinities were synthesized with an N-terminal CysTyr and C-terminal Lys4 sequence. Dissociation constants of these hybrid PNAs were determined by the Dynabead-based solution assay to be about 10 pM for the highest affinity binders. PMID- 16314304 TI - Glucose repression of the Escherichia coli sdhCDAB operon, revisited: regulation by the CRP*cAMP complex. AB - Expression of the Escherichia coli sdhCDAB operon encoding the succinate dehydrogenase complex is regulated in response to growth conditions, such as anaerobiosis and carbon sources. An anaerobic repression of sdhCDAB is known to be mediated by the ArcB/A two-component system and the global Fnr anaerobic regulator. While the cAMP receptor protein (CRP) and Cra (formerly FruR) are known as key mediators of catabolite repression, they have been excluded from the glucose repression of the sdhCDAB operon. Although the glucose repression of sdhCDAB was reported to involve a mechanism dependent on the ptsG expression, the molecular mechanism underlying the glucose repression has never been clarified. In this study, we re-examined the mechanism of the sdhCDAB repression by glucose and found that CRP directly regulates expression of the sdhCDAB operon and that the glucose repression of this operon occurs in a cAMP-dependent manner. The levels of phosphorylated enzyme IIA(Glc) and intracellular cAMP on various carbon sources were proportional to the expression levels of sdhC-lacZ. Disruption of crp or cya completely abolished the glucose repression of sdhC-lacZ expression. Together with data showing correlation between the intracellular cAMP concentrations and the sdhC-lacZ expression levels in several mutants and wild type, in vitro transcription assays suggest that the decrease in the CRP.cAMP level in the presence of glucose is the major determinant of the glucose repression of the sdhCDAB operon. PMID- 16314305 TI - Lack of MSH2 involvement differentiates V(D)J recombination from other non homologous end joining events. AB - V(D)J recombination and class switch recombination are the two DNA rearrangement events used to diversify the mouse and human antibody repertoires. While their double strand breaks (DSBs) are initiated by different mechanisms, both processes use non-homologous end joining (NHEJ) in the repair phase. DNA mismatch repair elements (MSH2/MSH6) have been implicated in the repair of class switch junctions as well as other DNA DSBs that proceed through NHEJ. MSH2 has also been implicated in the regulation of factors such as ATM and the MRN (Mre11, Rad50, Nbs1) complex, which are involved in V(D)J recombination. These findings led us to examine the role of MSH2 in V(D)J repair. Using MSH2-/- and MSH2+/+ mice and cell lines, we show here that all pathways involving MSH2 are dispensable for the generation of an intact pre-immune repertoire by V(D)J recombination. In contrast to switch junctions and other DSBs, the usage of terminal homology in V(D)J junctions is not influenced by MSH2. Thus, whether the repair complex for V(D)J recombination is of a canonical NHEJ type or a separate microhomology-mediated end joining (MMEJ) type, it does not involve MSH2. This highlights a distinction between the repair of V(D)J recombination and other NHEJ reactions. PMID- 16314306 TI - An autoregulatory loop controlling orphan nuclear receptor DAX-1 gene expression by orphan nuclear receptor ERRgamma. AB - The estrogen receptor-related receptor gamma (ERRgamma/ERR3/NR3B3) is a member of the nuclear receptor superfamily that activates transcription in the absence of ligand. However, the detailed mechanism of gene regulation by ERRgamma is not fully understood. In this study we have found that the orphan nuclear receptor ERRgamma activates the DAX-1 promoter, which, in turn, represses transactivation by ERRgamma. Serial deletions of mouse DAX-1 (mDAX-1) gene promoter have revealed that the region responding to ERRgamma is located between -129 and -121 bp and 334 and -326 bp. Gel shift assays and chromatin immunoprecipitation (ChIP) assays demonstrated that ERRgamma binds directly to the mDAX-1 promoter. Site-directed mutagenesis results demonstrated that ERRE1 (-129 to -121 bp) is more important than ERRE2 (-334 to -326 bp) which is not conserved in the human DAX-1 promoter. In addition, adenovirus-mediated overexpression of ERRgamma induced DAX-1 gene expression in MCF-7 breast cancer cells that co-expressed ERRgamma and DAX-1. Moreover, yeast two-hybrid and glutathione S-transferase (GST)-pull down assays demonstrated that DAX-1 physically interacted with ERRgamma and inhibited ERRgamma transactivation, and that this interaction was dependent on the AF-2 domain of ERRgamma. In addition, in vitro competition assays showed that DAX-1 inhibited PGC-1alpha mediated ERRgamma transactivation, via competition between these two factors for the AF-2 binding domain. We thus propose a novel autoregulatory loop that controls DAX-1 gene expression by ERRgamma. PMID- 16314307 TI - Footprinting of mammalian promoters: use of a CpG DNA methyltransferase revealing nucleosome positions at a single molecule level. AB - Promoters are molecular 'modules', which are controlled as individual entities yet are often analyzed by nuclease digestion methodologies which, a priori, destroy this modularity. About 40% of mammalian genes contain CpG islands in their promoters and exonic regions, which are normally unmethylated. We developed a footprinting strategy to map the chromatin structure at unmethylated CpG islands by treatment of isolated nuclei with the CpG-specific DNA methyltransferase SssI (M.SssI), followed by genomic bisulfite sequencing of individual progeny DNA molecules. This gave single molecule resolution over the promoter region and allowed for the physical linkage between binding sites on individual promoter molecules to be maintained. Comparison of the p16 promoters in two human cell lines, J82 and LD419, expressing the p16 gene at 25-fold different levels showed that the two cell lines contain remarkably different, heterogeneously positioned nucleosomes over the promoter region, which were not distinguishable by standard methods using nucleases. Our high resolution approach gives a 'digitized' visualization of each promoter providing information regarding nucleosome occupancy and may be utilized to define transcription factor binding and chromatin remodeling. PMID- 16314308 TI - Bis-aptazyme sensors for hepatitis C virus replicase and helicase without blank signal. AB - The fusion molecule (i.e. aptazyme) of aptamer and hammerhead ribozyme was developed as in situ sensor. Previously, the hammerhead ribozyme conjugated with aptamer through its stem II module showed a significant blank signal by self cleavage. To reduce or remove its self-cleavage activity in the absence of target molecule, rational designs were attempted by reducing the binding affinity of the aptazyme to its RNA substrate, while maintaining the ribonuclease activity of the aptazyme. Interestingly, the bis-aptazymes which comprise the two aptamer-binding sites at both stem I and stem III of the hammerhead ribozyme showed very low blank signals, and their ratios of reaction rate constants, i.e. signal to noise ratios, were several tens to hundred times higher than those of the stem II conjugated bis-aptazymes. The reduction in the blank signals seems to be caused by a higher dissociation constant between the main strand of the bis-aptazyme and its substrate arising from multi-point base-pairing of the bis-aptazymes. The bis aptazymes for HCV replicase and helicase showed high selectivity against other proteins, and a linear relationship existed between their ribozyme activities and the target concentrations. In addition, a bis-aptazyme of dual functions was designed by inserting both aptamers for HCV replicase and helicase into the stem I and stem III of hammerhead ribozyme, respectively, and it also showed greater sensitivity and specificity for both proteins without blank signal. PMID- 16314309 TI - Real-time quantification of microRNAs by stem-loop RT-PCR. AB - A novel microRNA (miRNA) quantification method has been developed using stem-loop RT followed by TaqMan PCR analysis. Stem-loop RT primers are better than conventional ones in terms of RT efficiency and specificity. TaqMan miRNA assays are specific for mature miRNAs and discriminate among related miRNAs that differ by as little as one nucleotide. Furthermore, they are not affected by genomic DNA contamination. Precise quantification is achieved routinely with as little as 25 pg of total RNA for most miRNAs. In fact, the high sensitivity, specificity and precision of this method allows for direct analysis of a single cell without nucleic acid purification. Like standard TaqMan gene expression assays, TaqMan miRNA assays exhibit a dynamic range of seven orders of magnitude. Quantification of five miRNAs in seven mouse tissues showed variation from less than 10 to more than 30,000 copies per cell. This method enables fast, accurate and sensitive miRNA expression profiling and can identify and monitor potential biomarkers specific to tissues or diseases. Stem-loop RT-PCR can be used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs). Furthermore, the concept of stem-loop RT primer design could be applied in small RNA cloning and multiplex assays for better specificity and efficiency. PMID- 16314310 TI - Multiplexed tandem PCR: gene profiling from small amounts of RNA using SYBR Green detection. AB - Multiplexed tandem PCR (MT-PCR) is a process for highly multiplexed gene expression profiling. In the first step, multiple primer pairs are added to the RNA to be analysed together with reverse transcriptase and Taq DNA polymerase. Following reverse transcription, the multiplexed amplicons are simultaneously amplified for a small number of cycles so as to avoid competition between amplicons. The reaction product is then diluted and analysed in multiple individual PCRs using primers nested inside the primers used for the multiplexed amplification. As the second PCR uses a template enriched in the amplicons of interest, the conditions can be optimized to significantly reduce 'primer dimer' formation allowing SYBR Green chemistry to be used for quantification. MT-PCR can be configured for as little as 10 pg RNA (equivalent to a single mammalian cell) and works well with RNA extracted from archival formalin-fixed paraffin-embedded sections. We illustrate MT-PCR with gene expression profiles of breast cancer cell lines. PMID- 16314311 TI - PCR inhibition by reverse transcriptase leads to an overestimation of amplification efficiency. AB - This study addresses the problem of PCR inhibition by reverse transcriptase. It has been shown that the inhibition occurs mostly when a small amount of RNA is taken for RT reaction, and it is more visible for rarely expressed transcripts. We show here that the inhibition takes place regardless of what amount of template is utilized for RT. The inhibition possesses a global nature, i.e. the amplification of any given transcript may be compromised with different levels of inhibition. The process of inhibition also explains wrongfully derived PCR amplification efficiencies, sometimes more than 100%, when the sequential dilutions of unpurified RT sample are utilized to build the calibration curve. The RT influences PCR not only by inhibiting it. When microgram(s) of RNA are taken for RT reaction, reverse transcriptase may cause overamplification of some transcripts under certain PCR conditions. The possible mechanism of RT influence on PCR is presented, and a purification method is implemented to remove the effects of RT on PCR. PMID- 16314312 TI - Gene identification in novel eukaryotic genomes by self-training algorithm. AB - Finding new protein-coding genes is one of the most important goals of eukaryotic genome sequencing projects. However, genomic organization of novel eukaryotic genomes is diverse and ab initio gene finding tools tuned up for previously studied species are rarely suitable for efficacious gene hunting in DNA sequences of a new genome. Gene identification methods based on cDNA and expressed sequence tag (EST) mapping to genomic DNA or those using alignments to closely related genomes rely either on existence of abundant cDNA and EST data and/or availability on reference genomes. Conventional statistical ab initio methods require large training sets of validated genes for estimating gene model parameters. In practice, neither one of these types of data may be available in sufficient amount until rather late stages of the novel genome sequencing. Nevertheless, we have shown that gene finding in eukaryotic genomes could be carried out in parallel with statistical models estimation directly from yet anonymous genomic DNA. The suggested method of parallelization of gene prediction with the model parameters estimation follows the path of the iterative Viterbi training. Rounds of genomic sequence labeling into coding and non-coding regions are followed by the rounds of model parameters estimation. Several dynamically changing restrictions on the possible range of model parameters are added to filter out fluctuations in the initial steps of the algorithm that could redirect the iteration process away from the biologically relevant point in parameter space. Tests on well-studied eukaryotic genomes have shown that the new method performs comparably or better than conventional methods where the supervised model training precedes the gene prediction step. Several novel genomes have been analyzed and biologically interesting findings are discussed. Thus, a self training algorithm that had been assumed feasible only for prokaryotic genomes has now been developed for ab initio eukaryotic gene identification. PMID- 16314313 TI - Synthesis of novel poly(dG)-poly(dG)-poly(dC) triplex structure by Klenow exo- fragment of DNA polymerase I. AB - The extension of the G-strand of long (700 bp) poly(dG)-poly(dC) by the Klenow exo(-) fragment of DNA polymerase I yields a complete triplex structure of the H DNA type. High-performance liquid chromatography analysis demonstrates that the length of the G-strand is doubled during the polymerase synthesis. Fluorescence resonance energy transfer analysis shows that the 5' ends of the G- and the C strands, labeled with fluorescein and TAMRA, respectively, are positioned close to each other in the product of the synthesis. Atomic force microscopy morphology imaging shows that the synthesized structures lack single-stranded fragments and have approximately the same length as the parent 700 bp poly(dG)-poly(dC). CD spectrum of the polymer has a large negative peak at 278 nm, which is characteristic of the poly(dG)-poly(dG)-poly(dC) triplex. The polymer is resistant to DNase and interacts much more weakly with ethidium bromide as compared with the double-stranded DNA. PMID- 16314314 TI - The excess of 5' introns in eukaryotic genomes. AB - In this work, 21 completely sequenced eukaryotic genomes were analyzed using an intragene comparison approach. We found that all of these genomes show a significant 5'-biased distribution of introns of protein-coding genes. Our findings are different from previous studies based on the intergene method, where introns are biased towards the 5' end of genes only in intron-poor genomes, but are evenly distributed in intron-rich genomes. In addition, by analyzing the patterns of intron distribution of a set of well-compiled housekeeping genes from human and their respective orthologs identified by a bidirectional best BLAST hit method from the other genomes, we found that the trend of 5'-biased intron positions of the set of housekeeping genes for each genome is much more skewed than that of all genes of the same genome, and rarely if any of the housekeeping genes examined have an extremely 3'-biased position distribution in which all introns of a gene are located only at the 3' portion of the gene. The most parsimonious explanation for our findings may be the model in which intron loss is caused by homologous recombination between the genomic copy of a gene and a reverse transcriptase product of a spliced mRNA. PMID- 16314315 TI - Activation of Sp1-mediated transcription by Rta of Epstein-Barr virus via an interaction with MCAF1. AB - Rta is a transcription factor encoded by BRLF1 of the Epstein-Barr virus (EBV). This factor is expressed during the immediate-early stage of the lytic cycle to activate the genes required for EBV lytic development. Although transcription activation by Rta is frequently associated with the binding of Rta to the Rta response element (RRE) in promoters, Rta sometimes activates promoters without an RRE. Here we show that Rta interacts with an Sp1-interacting protein, MBD1 containing chromatin-associated factor 1 (MCAF1). This interaction is critical to the formation of an Sp1-MCAF1-Rta complex at Sp1 sites. Therefore, following lytic induction and the expression of Rta, Rta increases Sp1-mediated transcription. The genes that are thus activated include p16, p21, SNRPN and BRLF1. However, the binding of Rta to RRE prevents the interaction between Rta and MCAF1; therefore, transcription activation by RRE depends only on Rta, and not on MCAF1 or Sp1. Furthermore, this study finds that MCAF1 promotes the expression of Rta and Zta from EBV, indicating that MCAF1 participates EBV lytic activation. Our study documents the critical role of Rta in regulating the transcription of the genes that are mediated by Sp1. PMID- 16314316 TI - Human Bex2 interacts with LMO2 and regulates the transcriptional activity of a novel DNA-binding complex. AB - Human Bex2 (brain expressed X-linked, hBex2) is highly expressed in the embryonic brain, but its function remains unknown. We have identified that LMO2, a LIM domain containing transcriptional factor, specifically interacts with hBex2 but not with mouse Bex1 and Bex2. The interaction was confirmed both by pull-down with GST-hBex2 and by coimmunoprecipitation assays in vivo. Using electrophoretic mobility shift assay, we have demonstrated the physical interaction of hBex2 and LMO2 as part of a DNA-binding protein complex. We have also shown that hBex2 can enhance the transcriptional activity of LMO2 in vivo. Furthermore, using mammalian two-hybrid analysis, we have identified a neuronal bHLH protein, NSCL2, as a novel binding partner for LMO2. We then showed that LMO2 could up-regulate NSCL2-dependent transcriptional activity, and hBex2 augmented this effect. Thus, hBex2 may act as a specific regulator during embryonic development by modulating the transcriptional activity of a novel E-box sequence-binding complex that contains hBex2, LMO2, NSCL2 and LDB1. PMID- 16314317 TI - SIP1/ZEB2 induces EMT by repressing genes of different epithelial cell-cell junctions. AB - SIP1/ZEB2 is a member of the deltaEF-1 family of two-handed zinc finger nuclear factors. The expression of these transcription factors is associated with epithelial mesenchymal transitions (EMT) during development. SIP1 is also expressed in some breast cancer cell lines and was detected in intestinal gastric carcinomas, where its expression is inversely correlated with that of E-cadherin. Here, we show that expression of SIP1 in human epithelial cells results in a clear morphological change from an epithelial to a mesenchymal phenotype. Induction of this epithelial dedifferentiation was accompanied by repression of several cell junctional proteins, with concomitant repression of their mRNA levels. Besides E-cadherin, other genes coding for crucial proteins of tight junctions, desmosomes and gap junctions were found to be transcriptionally regulated by the transcriptional repressor SIP1. Moreover, study of the promoter regions of selected genes by luciferase reporter assays and chromatin immunoprecipitation shows that repression is directly mediated by SIP1. These data indicate that, during epithelial dedifferentiation, SIP1 represses in a coordinated manner the transcription of genes coding for junctional proteins contributing to the dedifferentiated state; this repression occurs by a general mechanism mediated by Smad Interacting Protein 1 (SIP1)-binding sites. PMID- 16314318 TI - The PSEA promoter element of the Drosophila U1 snRNA gene is sufficient to bring DmSNAPc into contact with 20 base pairs of downstream DNA. AB - Most of the major spliceosomal small nuclear RNAs (snRNAs) (i.e. U1, U2, U4 and U5) are synthesized by RNA polymerase II (pol II). In Drosophila melanogaster, the 5'-flanking DNA of these genes contains two conserved elements: the proximal sequence element A (PSEA) and the proximal sequence element B (PSEB). The PSEA is essential for transcription and is recognized by DmSNAPc, a multi-subunit protein complex. Previous site-specific protein-DNA photo-cross-linking assays demonstrated that one of the subunits of DmSNAPc, DmSNAP43, remains in close contact with the DNA for 20 bp beyond the 3' end of the PSEA, a region that contains the PSEB. The current work demonstrates that mutation of the PSEB does not abolish the cross-linking of DmSNAP43 to the PSEB. Thus the U1 PSEA alone is capable of bringing DmSNAP43 into close contact with this downstream DNA. However, mutation of the PSEB perturbs the cross-linking pattern. In concordance with these findings, PSEB mutations result in a 2- to 4-fold reduction in U1 promoter activity when assayed by transient transfection. PMID- 16314319 TI - A thermodynamic model of transcriptome formation. AB - The genome supplies information on both the quality and quantity of the transcriptome. However, as it remains unknown how a cell determines transcript levels from the genome sequences, despite comprehensive knowledge of the cellular components involved, the quantity information held by the genome cannot as yet be derived from nucleotide sequences. The model presented here explains on a thermodynamic basis how the components decode the genome to form and maintain the transcriptome. The model describes the level of a transcript as a pseudo equilibrium between velocities of synthesis and degradation, both of which are controlled by sequence-specific interactions between protein factors and nucleic acids. Each of the transcript levels can be described by a single equation expressing a function of the activity concentrations of the protein factors. Quantitative information in the genome can thus be transformed into constants determined from the nucleotide sequences. Using this model, the transcriptome can be traced back to the protein factors and the state of chromosome packaging. The total description of transcript levels allows the model to be verified through comparison of derived hypotheses with comprehensive measurements of the transcriptome. The hypotheses thus derived in the present study are well supported by experimental microarray data, confirming the appropriateness of the model. PMID- 16314320 TI - A second look at cellular mRNA sequences said to function as internal ribosome entry sites. AB - This review takes a second look at a set of mRNAs that purportedly employ an alternative mechanism of initiation when cap-dependent translation is reduced during mitosis or stress conditions. A closer look is necessary because evidence cited in support of the internal initiation hypothesis is often flawed. When putative internal ribosome entry sequences (IRESs) are examined more carefully, they often turn out to harbor cryptic promoters or splice sites. This undermines the dicistronic assay, wherein IRES activity is measured by the ability to support translation of the 3' cistron. Most putative IRESs still have not been checked carefully to determine whether the dicistronic vector produces only the intended dicistronic mRNA. The widespread use of the pRF vector is a major problem because this vector, which has Renilla luciferase as the 5' cistron and firefly luciferase as the 3' cistron, has been found to generate spliced transcripts. RNA transfection assays could theoretically circumvent these problems, but most candidate IRESs score very weakly in that test. The practice of calling even very weak results 'positive' is one of the problems discussed herein. The extremely low efficiency of putative IRESs is inconsistent with their postulated biological roles.' PMID- 16314321 TI - Structure-specific binding of MeCP2 to four-way junction DNA through its methyl CpG-binding domain. AB - MeCP2, whose methylated DNA-binding domain (MBD) binds preferentially to DNA containing 5Me-CpG relative to linear unmethylated DNA, also binds preferentially, and with similar affinity, to unmethylated four-way DNA junctions through the MBD. The Arg133Cys (R133C) mutation in the MBD, a Rett syndrome mutation that abolishes binding to methylated DNA, leads to only a slight reduction in the affinity of the MBD for four-way junctions, suggesting distinct but partially overlapping modes of binding to junction and methylated DNA. Binding to unmethylated DNA junctions is likely to involve a subset of the interactions that occur with methylated DNA. High-affinity, methylation independent binding to four-way junctions is consistent with additional roles for MeCP2 in chromatin, beyond recognition of 5Me-CpG. PMID- 16314322 TI - Rotation of DNA around intact strand in human topoisomerase I implies distinct mechanisms for positive and negative supercoil relaxation. AB - Topoisomerases are enzymes of quintessence to the upkeep of superhelical DNA, and are vital for replication, transcription and recombination. An atomic-resolution model for human topoisomerase I in covalent complex with DNA is simulated using molecular dynamics with external potentials that mimic torque and bias the DNA duplex downstream of a single-strand cut to rotate around the intact strand, according to the prevailing enzymatic mechanism. The simulations reveal the first dynamical picture of how topoisomerase accommodates large-scale motion of DNA as it changes its supercoiling state, and indicate that relaxation of positive and negative supercoils are fundamentally different. To relax positive supercoils, two separate domains (the 'lips') of the protein open up by about 10-14 A, whereas to relax negative supercoils, a continuous loop connecting the upper and lower parts (and which was a hinge for opening the lips) stretches about 12 A while the lips remain unseparated. Normal mode analysis is additionally used to characterize the functional flexibility of the protein. Remarkably, the same combination of low-frequency eigenvectors exhibit the dominant contribution for both rotation mechanisms through a see-saw motion. The simulated mechanisms suggest mutations to control the relaxation of either type of supercoiling selectively and advance a hypothesis for the debated role of the N-terminal domain in supercoil relaxation. PMID- 16314323 TI - AP endonuclease deficiency results in extreme sensitivity to thymidine deprivation. AB - Thymidine depletion is toxic to virtually all actively growing cells. The fundamental mechanism responsible for thymidineless death remains unknown. One event thought to be critical in causing the toxicity of thymidine depletion is a sharp rise in the ratio of dUTP to dTTP and subsequent incorporation of dUTP into DNA. Maneuvers to alter dUTP levels appear to alter the toxicity of thymidine depletion. However, loss of uracil-DNA-N-glycosylase activity does not appear to change the toxicity of thymidine deprivation significantly. This study proposes to define the role of uracil base excision repair (BER) in mediating thymidineless death. The toxicity of thymidine deprivation induced by the antifolate aminopterin was measured in a series of mutant Saccharomyces cerevisiae strains deficient in various steps in uracil-BER. Most mutants displayed modest changes in their sensitivity to aminopterin, with the exception of cells lacking the abasic endonuclease Apn1. apn1 mutants displayed a profound sensitivity to aminopterin that was relieved in an apn1 ung1 double mutant. Wild type and apn1 mutants displayed similar levels of DNA damage and S-phase arrest during aminopterin treatment. A significant portion of cell killing occurred after removal of aminopterin in both wild-type and apn1 mutant cells. apn1 mutants showed a complete inability to re-initiate DNA replication following removal of aminopterin. These findings suggest recovery from arrest is a crucial step in determining the response to thymidine deprivation and that interruptions in uracil-BER increase the toxicity of thymidine deprivation by blocking re initiation of replication rather than inciting global DNA damage. Inhibition of apurinic/apyrimidinic endonuclease may therefore be a reasonable approach to increase the efficacy of anticancer chemotherapies based on thymidine depletion. PMID- 16314324 TI - A retrocopy of a gene can functionally displace the source gene in evolution. AB - The e(y)2 gene of Drosophila melanogaster encodes the ubiquitous evolutionarily conserved co-activator of RNA polymerase II that is involved in transcription regulation of a high number of genes. The Drosophila e(y)2b gene, paralogue of the e(y)2 has been found. The analysis of structure of the e(y)2, e(y)2b and its orthologues from other species reveals that the e(y)2 gene derived as a result of retroposition of the e(y)2b during Drosophila evolution. The mRNA-derived retrogenes lack introns or regulatory regions; most of them become pseudogenes whereas some acquire tissue-specific functions. Here we describe the different situation: the e(y)2 retrogene performs the general function and is ubiquitously expressed, while the source gene is functional only in a small group of male germ cells. This must have resulted from retroposition into a transcriptionally favorable region of the genome. PMID- 16314325 TI - DNA end-directed and processive nuclease activities of the archaeal XPF enzyme. AB - The XPF/Mus81 family of structure-specific nucleases cleaves branched or nicked DNA substrates and are implicated in a wide range of DNA repair and recombination processes. The structure of the crenarchaeal XPF bound to a DNA duplex has revealed a plausible mechanism for DNA binding, involving DNA distortion into upstream and downstream duplexes engaged by the two helix-hairpin-helix domains that form a dimeric structure at the C-terminus of the enzyme. A flexible linker joins these to the dimeric nuclease domain, and a C-terminal motif interacts with the sliding clamp, which is essential for the activity of the enzyme. Here, we demonstrate the importance of the downstream duplex in directing the endonuclease activity of crenarchaeal XPF, which is similar to that of Mus81-Eme1, and suggest a mechanistic basis for this control. Furthermore, our data reveal that the enzyme can digest a nicked DNA strand processively over at least 60 nt in a 3'-5' direction and can remove varied types of DNA lesions and blocked DNA termini. This in vitro activity suggests a potential role for crenarchaeal XPF in a variety of repair processes for which there are no clear pathways in archaea. PMID- 16314326 TI - Protecting workers in licensed premises from the effects of secondhand smoke. PMID- 16314329 TI - Prevention of occupational asthma--practical implications for occupational physicians. AB - BACKGROUND: Occupational factors have been estimated to contribute to approximately 10% of adult-onset asthma and occupational asthma (OA) is one of the most common occupational lung diseases in industrialized areas. Persistent asthma frequently occurs with significant socio-economic impacts. METHODS: A literature search was performed using PubMed. The key term searched was occupational asthma combined with prevention. RESULTS: Primary prevention has been effective for OA related to natural rubber latex, and may have reduced the incidence of diisocyanate-induced asthma. Medical health surveillance has been effective in settings such as the detergent enzyme industry, workers exposed to complex platinum salts and likely for diisocyanate workers in Ontario. Tertiary prevention is still required for workers with OA and can improve prognosis. CONCLUSIONS: OA is potentially preventable. Sufficient studies have demonstrated the rationale and benefit of primary preventive strategies. Medical health surveillance programs combined with occupational hygiene measures and worker education have been associated with improved outcomes but further studies are needed to understand the optimum frequency and measures for such programs and to identify the separate contribution of the components. Until primary and secondary prevention is better understood and implemented, there will also remain a need for tertiary preventive measures. PMID- 16314330 TI - Concerns for asthma at pre-placement assessment and health surveillance in platinum refining--a personal approach. AB - Platinum, a noble metal, is inert in the body and has an important use in medical applications. It is used in autocatalysts to control harmful vehicular emissions and the catalytic effects improve efficiencies of pharmaceutical and petrochemical processes. The refining process involves exposure to halogeno complex salts of platinum which are potent allergens. They induce symptoms typical of a type I allergy, the most significant of which is asthma. Platinum refining not only exposes employees to the risk of sensitization to these salts but also to respiratory irritants. Inhalation of these aggravates pre-existing asthma. The increasing incidence of asthma in the community requires that prospective employees for platinum refining be assessed carefully to establish their respiratory health status in relation to the risk of sensitization or aggravation of pre-existing asthma. Routine medical surveillance has been shown to reduce persistence of asthma in sensitized workers who cease exposure to the platinum salts upon diagnosis. Skin prick test using dilute platinum salt solutions can detect sensitization at an early stage and this has become the mainstay of surveillance programmes as it is objective and reproducible as well as predictive for the development of symptoms when exposure is allowed to continue. Symptoms are not sensitive or specific. Smoking is a significant risk factor. PMID- 16314331 TI - Occupational asthma prevention and management in industry--an example of a global programme. AB - Isocyanates are widely used in the manufacture of polyurethane foams, plastics, coatings and adhesives, and are known to cause occupational asthma in a proportion of exposed workers. Substitution as a prevention strategy is not currently a feasible option. For this reason, health and safety professionals working together in an automotive coatings business created a proactive global programme to address the known potential effects of isocyanates on its workers. The goals of the programme are prevention, early detection and mitigation of effect of key endpoints, especially asthma and to a lesser degree dermatitis, in people who are occupationally exposed, or potentially exposed, to isocyanates and products containing isocyanates. The surveillance programme for isocyanates has several important components, which include assessment of exposure, pre-placement questionnaire and spirometry, training and education of employees, regularly administered periodic questionnaires, medical assessment for abnormal questionnaire responses, process for early reporting and investigation of symptomatic employees and incidents, group data review and management reporting. Although regional differences exist regarding availability of specialized services, we have successfully implemented this programme in parts of North America, Europe and Latin America, and are currently implementing elsewhere. These simple control measures are relatively inexpensive and can be applied in even small business settings. It is recommended that all employers who manufacture, handle or use isocyanate-containing products consider such a strategy. PMID- 16314332 TI - Future advances in work-related asthma and the impact on occupational health. AB - Work-related asthma (WRA) and WRA-like conditions refer to all situations in which asthma or asthma-like syndromes occur or worsen in the workplace. This occurs in approximately 10% of all adult-onset asthma subjects. Occupational asthma (OA) is a specific type of WRA that is 'caused' by the workplace, being mediated either by an allergic process with a latency period or by a non-allergic irritant-induced mechanism. Personal asthma can also 'worsen' at work (work aggravated or exacerbated asthma), the reasons, mechanisms, extent and consequences of this situation being unknown. The author reviews various aspects of WRA with an emphasis on OA (about which more is known) and proposes key issues that need to be further studied, proposed and applied in at-risk workplaces in order to improve recognition, diagnosis and management of this condition. OA represents a unique situation that, unfortunately, is only very rarely provided to health-care providers: affected workers can be cured with minimal impact on quality of life. All efforts should be made to achieve this goal at an affordable socio-economic cost. PMID- 16314333 TI - Saved by a material safety data sheet. AB - BACKGROUND: We present the case of a young female laboratory worker who developed acute hepatic encephalopathy. OBJECTIVE: To show that knowledge of occupational exposures to causative agents can alter therapeutic management. METHODS: Although the patient was in a deep coma, her family members examined the workplace material safety data sheet, revealing exposure to chloroform. Since most chemical induced hepatitis is self-limiting, a scheduled liver transplantation was postponed. RESULTS: The patient recovered. Subsequent air sampling suggested that the patient had been exposed to chloroform at a concentration of more than 15 ppm for 2 weeks. CONCLUSION: Our case report demonstrates the importance of obtaining an occupational history and how the patient's family can be important in this process. PMID- 16314334 TI - Metal fume fever presenting as aseptic meningitis with pericarditis, pleuritis and pneumonitis. AB - BACKGROUND: Metal fume fever (MFF) is a well-known complication of zinc oxide fume inhalation. Prompt recognition of this condition is essential for the proper medical management of this self-limited disease. AIM: To present a unique and unusual case of MFF. RESULTS: Our patient is a 25-year-old male welder who had MFF and presented with aseptic meningitis with pericarditis, pleuritis and pneumonitis. To our knowledge, this is the first case of MMF presenting with these signs and symptoms. CONCLUSIONS: MFF can present with a systemic inflammatory response causing a multi-organ serositis. Our case highlights the utmost importance of obtaining an occupational history on all our patients, even if they are critically ill. PMID- 16314336 TI - Patient-related factors of compliance in heart failure: some new insights into an old problem. PMID- 16314337 TI - Mechanisms of death in elderly patients with acute myocardial infarction exposed to fibrinolytic therapy. PMID- 16314338 TI - The clinical impact of implantable loop recorders in patients with syncope. AB - AIMS: Implantable loop recorders (ILR) provide an opportunity to record ECG data from a spontaneous syncopal event. We conducted a randomized study to investigate the impact of the Reveal Plus ILR on an unselected population of patients with recurrent syncope. Initial follow-up (at least 6 months) did not demonstrate a reduction in syncopal events or an improvement in quality of life. We report the planned extension of follow-up to 18 months. METHODS AND RESULTS: All patients presenting acutely with recurrent unexplained syncope over a 16-month period, following a basic clinical work-up, were randomized to receive the ILR or conventional investigation and management. A total of 421 patients presented, 201 were eligible, median age 74, (IQ range 61-81) 54% female, with median syncopes 3 (IQ range 2-6). Median follow-up 17 months (IQ range 9-23). 42 (43%) of ILR patients and 8 (6%) of conventional patients received an ECG diagnosis (hazard ratio 6.53, 95% CI 3.73-11.4, P<0.001). Time to second syncope was significantly longer for ILR patients, although of borderline significance (P=0.04). A greater variety of diagnoses and treatments were seen in ILR patients. ILR patients had fewer post-randomization investigations and fewer days in hospital; however, cost savings were not statistically significant. There was improved quality of life in the ILR group (visual analogue scales, P=0.03) for general wellbeing. Overall mortality was 12% with no difference between the two groups. CONCLUSION: Investigation by the ILR significantly increases the diagnostic rate and ECG directed treatments in a typical unselected syncopal population. Long-term follow up has demonstrated a significant subsequent reduction in syncopal events with improved quality of life. PMID- 16314339 TI - Idiopathic retroperitoneal fibrosis: a rare onset of the illness caused by haemorrhagic fever with renal syndrome. PMID- 16314340 TI - The cucurbit images (1515-1518) of the Villa Farnesina, Rome. AB - BACKGROUND: The gorgeous frescoes organized by the master Renaissance painter Raphael Sanzio (1483-1520) and illustrating the heavenly adventures of Cupid and Psyche were painted between 1515 and 1518 to decorate the Roman villa (now known as the Villa Farnesina) of the wealthy Sienese banker Agostino Chigi (1466-1520). Surrounding these paintings are festoons of fruits, vegetables and flowers painted by Giovanni Martini da Udine (1487-1564), which include over 170 species of plants. A deconstruction and collation of the cucurbit images in the festoons makes it possible to evaluate the genetic diversity of cucurbits in Renaissance Italy 500 years ago. FINDINGS: The festoons contain six species of Old World cucurbits, Citrullus lanatus (watermelon), Cucumis melo (melon), Cucumis sativus (cucumber), Ecballium elaterium (squirting cucumber), Lagenaria siceraria (bottle gourd) and Momordica balsamina (balsam apple), and two or three species of New World cucurbits, Cucurbita maxima, C. pepo and, perhaps, C. moschata (pumpkin, squash, gourd). The images of C. maxima are the first illustrations of this species in Europe. PMID- 16314341 TI - Levels of H-ras codon 61 CAA to AAA mutation: response to 4-ABP-treatment and Pms2-deficiency. AB - DNA mismatch repair (MMR) deficiencies result in increased frequencies of spontaneous mutation and tumor formation. In the present study, we tested the hypothesis that a chemically-induced mutational response would be greater in a mouse with an MMR-deficiency than in the MMR-proficient mouse models commonly used to assay for chemical carcinogenicity. To accomplish this, the induction of H-ras codon 61 CAA-->AAA mutation was examined in Pms2 knockout mice (Pms2-/-, C57BL/6 background) and sibling wild-type mice (Pms2+/+). Groups of five or six neonatal male mice were treated with 0.3 micromol 4-aminobiphenyl (4-ABP) or the vehicle control, dimethylsulfoxide. Eight months after treatment, liver DNAs were isolated and analysed for levels of H-ras codon 61 CAA-->AAA mutation using allele-specific competitive blocker-PCR. In Pms2-proficient and Pms2-deficient mice, 4-ABP treatment caused an increase in mutant fraction (MF) from 1.65x10(-5) to 2.91x10(-5) and from 3.40x10(-5) to 4.70x10(-5), respectively. Pooling data from 4-ABP-treated and control mice, the approximately 2-fold increase in MF observed in Pms2-deficient as compared with Pms2-proficient mice was statistically significant (P=0.0207) and consistent with what has been reported previously in terms of induction of G:C-->T:A mutation in a Pms2-deficient background. Pooling data from both genotypes, the increase in H-ras MF in 4-ABP treated mice, as compared with control mice, did not reach the 95% confidence level of statistical significance (P=0.0606). The 4-ABP treatment caused a 1.76 fold and 1.38-fold increase in average H-ras MF in Pms2-proficient and Pms2 deficient mice, respectively. Furthermore, the levels of induced mutation in Pms2 proficient and Pms2-deficient mice were nearly identical (1.26x10(-5) and 1.30x10(-5), respectively). We conclude that Pms2-deficiency does not result in an amplification of the H-ras codon 61 CAA-->AAA mutational response induced by 4 ABP. PMID- 16314342 TI - DNA damage checkpoints in mammals. AB - DNA damage is a common event and probably leads to mutation or deletion within chromosomal DNA, which may cause cancer or premature aging. DNA damage induces several cellular responses including DNA repair, checkpoint activity and the triggering of apoptotic pathways. DNA damage checkpoints are associated with biochemical pathways that end delay or arrest of cell-cycle progression. These checkpoints engage damage sensor proteins, such as the Rad9-Rad1-Hus1 (9-1-1) complex, and the Rad17-RFC complex, in the detection of DNA damage and transduction of signals to ATM, ATR, Chk1 and Chk2 kinases. Chk1 and Chk2 kinases regulate Cdc25, Wee1 and p53 that ultimately inactivate cyclin-dependent kinases (Cdks) which inhibit cell-cycle progression. In this review, we discuss the molecular mechanisms by which DNA damage is recognized by sensor proteins and signals are transmitted to Cdks. We classify the genes involved in checkpoint signaling into four categories, namely sensors, mediators, transducers and effectors, although their proteins have the broad activity, and thus this classification is for convenience and is not definitive. PMID- 16314343 TI - Expression of cyclooxygenase-2 and matrix metalloproteinase-9 in gastric carcinoma and its correlation with angiogenesis. AB - OBJECTIVE: To investigate the expression of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-9 in gastric carcinomas, and to correlate this expression with clinicopathological parameters and angiogenesis. METHODS: Ninety-six resected tumor specimens from patients with gastric carcinoma were obtained, and 30 corresponding paracancerous normal tissues were randomly selected as a control. Immunohistochemical staining was used for detecting the expression of COX-2 and MMP-9. Monoclonal antibody against CD34 was used for displaying vascular endothelial cells, and microvascular density (MVD) was calculated by counting of CD34-positive vascular endothelial cells. RESULTS: The positive expression rates of COX-2, MMP-9 and MVD in the cancerous tissue were 80.2%, 74.0%, and 32.5 +/- 8.3, respectively, which were significantly higher than those in the normal tissue (P < 0.01). COX-2, MMP-9 expression rates and MVD in the patients with stages III and IV were 91.4%, 84.5% and 34.9 +/- 8.7, respectively, which were significantly higher than those in the patients with stages I and II (P < 0.01). In addition, the Spearman rank correlation test showed that tumor MVD was closely associated with COX-2 (r = 0.311, P < 0.01) and MMP-9 (r = 0.349, P < 0.01) expressions. CONCLUSIONS: Overexpression of COX-2 and MMP-9 is related to tumor invasion and lymph node metastasis in the gastric carcinoma. These results provide evidence that COX-2 contribute to gastric cancer development by promoting MMP-9 expression and angiogenesis. PMID- 16314344 TI - Clinicopathologic and prognostic significance of MMP-7 (matrilysin) expression in human rectal cancer. AB - OBJECTIVE: There is increasing evidence to indicate that MMP-7 plays a more important role in tumor progression than other MMPs. The aim of this study was to detect MMP-7 expression in human rectal cancer and normal rectal tissue and to determine whether it is correlated with invasion and metastasis of human rectal cancer. METHODS: Eighty-six paired samples of rectal cancer and distant normal rectal tissue obtained from 100 inpatients were allocated into two groups (cancer group and control group). MMP-7 mRNA was detected by relative quantitative real time RT-PCR and MMP-7 protein was examined by immunohistochemical staining and computerized image analysis. RESULTS: MMP-7 mRNA expression in cancer group was higher than that in control group (P = 0.006), the expression ratios of 31 samples (37.35%) were <1 and 52 (62.65%) were >1. The mRNA expression level was correlated with Dukes Staging, histological differentiation grade and CEA level. The MMP-7 protein expression was in accordance with mRNA expression level. The positive degree of immunohistochemical staining in cancer group (1.82 +/- 0.03) was different from that in control group (1.17 +/- 0.13, P = 0.002). Moreover, in cancer group the positive staining degree in high-level mRNA cancers (2.04 +/- 0.18, n = 52) was higher than that in low-level mRNA ones (1.58 +/- 0.23, n = 31, P = 0.008). CONCLUSIONS: Our results suggest that MMP-7 plays an important role in the progression of human rectal cancer. MMP-7 may be selected as a clinical diagnosis and prognosis index in rectal cancer. PMID- 16314345 TI - Phase II feasibility study of high-dose radiotherapy for prostate cancer using proton boost therapy: first clinical trial of proton beam therapy for prostate cancer in Japan. AB - OBJECTIVE: To assess the feasibility of high-dose radiotherapy for prostate cancer using proton boost therapy following photon radiotherapy. METHODS: The primary endpoint was acute grade 3 or greater genitourinary (GU) and gastrointestinal (GI) toxicities. The study included patients with clinical stage T1-3N0M0 prostate cancer. Radiotherapy consisted of 50 Gy/25 fx photon irradiation to the prostate and the bilateral seminal vesicles followed by proton boost of 26 Gy(E)/13 fx to the prostate alone. Hormonal therapy was allowed before and during the radiation therapy. RESULTS: Between January 2001 and January 2003, 30 patients were enrolled in this study. Acute grade 1/2 GU and GI toxicities were observed in 20/4 and 17/0 patients, respectively. With the median follow-up period of 30 months (range 20-45), late grade 1/2 GU and GI toxicities occurred in 2/3 and 8/3 patients, respectively. No grade 3 or greater acute or late toxicities were observed. All patients were alive, but six patients relapsed biochemically after 7-24 months. CONCLUSIONS: Proton boost therapy following photon radiotherapy for prostate cancer is feasible. To evaluate the efficacy and safety of proton beam therapy, a multi-institutional phase II trial is in progress in Japan. PMID- 16314347 TI - Comment on: Evidence-based review of antifungal prophylaxis in neutropenic patients with haematological malignancies. PMID- 16314348 TI - Management of conflict of interest issues in the activities of the American Heart Association Emergency Cardiovascular Care Committee, 2000-2005. PMID- 16314349 TI - Major changes in the 2005 AHA Guidelines for CPR and ECC: reaching the tipping point for change. PMID- 16314375 TI - 2005 American Heart Association Guidelines for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care. PMID- 16314376 TI - TRPV1 gene knockout impairs postischemic recovery in isolated perfused heart in mice. AB - BACKGROUND: Although pharmacological studies suggest that the transient receptor potential vanilloid type 1 (TRPV1) channels expressed in sensory nerve fibers innervating the heart may exert a cardioprotective effect, definitive evidence supporting such a notion is lacking. In addition, function and regulation of sensory neuropeptides, namely, calcitonin gene-related peptide (CGRP) and substance P (SP), in the face of challenges induced by cardiac injury in the presence or absence of the TRPV1 are largely unknown. METHODS AND RESULTS: The hearts of gene-targeted TRPV1-null mutant (TRPV1(-/-)) mice or wild-type (WT) mice were perfused in a Langendorff apparatus in the presence or absence of capsazepine (a TRPV1 receptor antagonist), CGRP, CGRP(8-37) (a CGRP receptor antagonist), SP, or RP67580 (a neurokinin-1 [NK1] receptor antagonist) when hearts were subjected to 40 minutes of ischemia and 30 minutes of reperfusion. Hemodynamic alterations and SP release measured by radioimmunoassay were assessed before and after ischemia/reperfusion injury of the heart. Expression of the NK1 receptor in the hearts of TRPV1(-/-) and WT mice were determined with the use of Western blot analyses. Impairment of postischemic recovery, defined by increased left ventricular end-diastolic pressure (LVEDP) and decreased left ventricular developed pressure (LVDP) and coronary flow (CF), was more severe in TRPV1(-/-) hearts than in WT hearts. Although it had no effect on postischemic recovery of TRPV1(-/-) hearts, blockade of the TRPV1 with capsazepine caused a most severe impairment of postischemic recovery in WT hearts compared with untreated WT and TRPV1(-/-) hearts. Exogenous CGRP and SP produced a significant improvement in postischemic recovery in both TRPV1(-/-) and WT hearts, and the maximal functional improvement in TRPV1(-/-) hearts was not different from that of WT hearts except that SP-induced increases in LVDP were larger in the former than in the latter. Blockade of the NK1 receptor with RP67580, but not blockade of the CGRP receptor with CGRP(8-37), caused more severe impairment in postischemic recovery in both TRPV1(-/-) and WT hearts than in untreated hearts in both genotypes. The release of SP after ischemia/reperfusion injury was increased in both WT and TRPV1(-/-) hearts, albeit with a smaller magnitude of the increase in the latter. Capsazepine attenuated injury-induced SP release in WT but not TRPV1( /-) hearts. There was no difference in the expression of the NK1 receptor between the 2 genotype hearts. CONCLUSIONS: Thus, our data show that (1) TRPV1 gene deletion decreases injury-induced SP release and impairs cardiac recovery function after ischemia/reperfusion injury; (2) TRPV1 gene deletion leads to reconditioning of the heart with improved postischemic recovery compared with that induced by acute TRPV1 blockade and in terms of cardiac response to exogenous SP; and (3) blockade of the NK1 but not CGRP receptors worsens postischemic recovery of hearts in both genotypes. Taken together, these data indicate that TRPV1 plays a role in protecting the heart from injury possibly via increasing SP release and that deletion of this receptor reconditions the heart for escaping, at least in part, from injury possibly via enhancing NK1 receptor function. PMID- 16314377 TI - Motherhood plans among young Australian women: who wants children these days? AB - Fertility rates in the developed world have been below replacement level for 25 years, and it is often assumed that this results from deliberate, unconstrained individual choice. Data from 7448 childless women aged 22 to 27, participating in the Australian Longitudinal Study on Women's Health, indicate that 9 per cent aspire to childlessness, with 72 per cent wanting 1 or 2 children and 19 per cent more. Differences in psychological functioning disappear after adjustment for socioeconomic variables. Women wanting 1 or 2 children also want paid work, while those wanting many children generally have traditional aspirations. Policy-makers need to consider strategies that support women to negotiate motherhood and paid work. PMID- 16314378 TI - Do psychological characteristics explain socioeconomic stratification of self rated health? AB - This study evaluated whether negative emotions explain socioeconomic status (SES) stratification of self-rated health (SRH) and whether this putative relation is independent of established SRH determinants. Mood disorders, trait negative affect and health status indices were assessed in a representative cross sectional survey of 3032 adults in the National Survey of Midlife Development in the United States (MIDUS). Adjustment for health behaviors and health status appreciably reduced SES influence on SRH, but adjustment for negative emotions did not. However, both psychological resources (e.g. social support, extraversion) and negative emotions independently predicted SRH. Detection of SRH determinants was sensitive to binary versus ordinal SRH definitions. PMID- 16314379 TI - Understanding and modeling health behavior: the multi-stage model of health behavior change. AB - The purpose of this study was to evaluate the Multi-Stage Model (MSM). The MSM proposes eight different stages that are distinguished by cognitions and behavior and concepts of habituation as well. The MSM stages were assessed in 835 rehabilitation patients with a questionnaire. People in the different stages were compared and trends across the means of individuals in the different stages were evaluated to test for discontinuity patterns. Discontinuity patterns across the stages were found in social-cognitive variables such as intention, self-efficacy, outcome expectancies and social support. Most predictions according to stage differences were empirically found. This supported the validity of the MSM. Implications for further investigations and stage-specific interventions are discussed. PMID- 16314380 TI - Autonomy and control: augmenting the validity of the theory of planned behaviour in predicting exercise. AB - This study examined the utility of the theory of planned behaviour (TPB) along with additional constructs in predicting exercise, and explored the motivational antecedents of exercise intentions. Participants included 162 Canadian University College students (61% females). Measures of TPB, autonomous and controlling intention, perceived autonomy support and core autonomous intention were completed during phase 1 of data collection. Two and three weeks later behaviour was assessed. Hierarchical regression analyses revealed that: (a) attitude and perceived behavioural control significantly predicted TPB intention and core autonomous intention; (b) subjective norm predicted controlling intention; and (c) perceived autonomy support predicted autonomous and core autonomous intention. TPB intention significantly predicted behaviour. TPB is a fairly useful model for predicting behaviour and important information can be gained when other measures of intention are explored. PMID- 16314381 TI - Outcome expectations and physical activity participation in two samples of older women. AB - Outcome expectations have received little study in physical activity (PA) research. This study examined whether initial outcome expectations and their achievement at 6 months (i.e. outcome realizations) predicted subsequent PA participation (7-12 months) in 118 older women. Initial outcome expectations were not associated with PA participation. Outcome realizations at 6 months, however, predicted subsequent PA participation (p < .05). Women with high expectations but low attainment had the lowest subsequent participation rates. Women with high attainment, regardless of expectations, had the highest rates. Findings replicate and extend an earlier study and argue for a more dynamic conceptualization of outcome expectations. PMID- 16314382 TI - (De)constructing body image. AB - The reification of body image leads to unarticulated ideological and conceptual assumptions that obscure the most dynamic and productive features of the construct. These assumptions are that body image: (1) 'exists'; (2) is a socially mediated product of perception; (3) is 'internal' and 'of the individual'; (4) can be treated and measured as if real; and (5) individuals' respond to body image measures as if neutrally providing information about pre-existing images held in their heads. We argue that it is more useful to consider body imaging as a process, an activity rather than a product. PMID- 16314383 TI - Physician and patient perceptions of the physician explanations in medical encounters. AB - Although, in actual practice, physicians have to subjectively judge the level of the explanation that they provide, little is known about this judgment. Therefore, making use of 630 physician-patient pairs in Japan, we investigated the association between patient and physician evaluations of physician explanations of medical test results and diagnoses. We found that the physician's judgment does not always agree with that of the patient, with regard to the level of explanation necessary. In addition, we first identified factors relating to the accuracy of physician judgments with regard to their explanations to patients. More studies will be necessary to verify the present findings. PMID- 16314384 TI - Coping with patients with medically unexplained symptoms: work-related strategies of physicians in primary health care. AB - General practitioners (GPs) often meet patients with medically unexplained symptoms (MUS). From a patient perspective, MUS is a well-acknowledged problem within the primary health care services today, but less is known about the GPs' perceptions. This study aims to elucidate GPs' perceptions of patients with MUS, focusing on stressing situations, emotional reactions and coping strategies. Twenty-seven physicians participated in focus-group discussions. In the analysis, where a phenomenographic approach was used, six situations were identified as being especially stressful in the encounter with these patients. The GPs described how they used both problem-focused and emotion-focused strategies, but with emotion-focused strategies slightly dominating, indicating that the GPs had difficulties in managing their own stress when working with patients with MUS. PMID- 16314385 TI - Exploring patient perceptions of movement through the stages of change model within a diabetes tele-care intervention. AB - Achieving strict control of diabetes is challenging for many patients and most need regularly to depend on ongoing support from health care professionals. One approach that might successfully provide this in a cost-effective and acceptable way is the delivery of advice and support over the telephone (tele-care). To date the process and effects of tele-care have however rarely been studied. This article explores the potential of one behavioural model, the trans-theoretical stages of change model (TTM), to understand and theorize about behaviour change for this chronic disease. Case study data are drawn from a wider randomized controlled trial (RCT) set up to assess the effectiveness of a pro-active call centre-based stepped treatment support to people with type 2 diabetes. The case study demonstrates the relevance and importance of the TTM model in this area. Different experiential and behavioural processes were of particular significance and held different meanings for the various change groups. PMID- 16314386 TI - A prospective study of the effects of psychological resources and depression in essential hypertension. AB - This study examined the influence of perceived health competence (PHC) and coping strategies in hypertension. Depressive mood state and dimensions of quality of life (QOL) (social support, satisfaction, well-being and free time) were also considered. One hundred and fifty participants were surveyed cross-sectionally and prospectively. Regression analysis revealed that the main predictors of the dimensions of QOL were PHC and depressive mood state. Emotional coping negatively predicted well-being. Unexpectedly, task-focused coping was unrelated to QOL domains. Longitudinal analyses did not support these relations. The data suggest that, in hypertension, PHC constitutes an important resource, whereas depressive mood state deteriorates QOL. PMID- 16314387 TI - The bases of goal setting in the self-regulation of hypertension. AB - We apply a new methodology to investigate goal setting by hypertensive patients that uncovers the reasons why people have a goal to manage hypertension or not (e.g. to reduce/maintain one's current blood pressure). The reasons are found to consist of superordinate goals in support of one's focal hypertension goal and the hierarchical mental network underlying the superordinate goals. We show that, not only do such superordinate goals influence patients' beliefs, feelings and decisions, but the relationships among superordinate goals are particularly efficacious in the formation of beliefs and attitudes, as well as intentions to self-regulate hypertension and actual efforts in doing so. Hypotheses were tested on a sample of 219 patients at a university-based hypertension clinic. PMID- 16314388 TI - A complex of two centrosomal proteins, CAP350 and FOP, cooperates with EB1 in microtubule anchoring. AB - The anchoring of microtubules (MTs) to subcellular structures is critical for cell shape, polarity, and motility. In mammalian cells, the centrosome is a prominent MT anchoring structure. A number of proteins, including ninein, p150Glued, and EB1, have been implicated in centrosomal MT anchoring, but the process is far from understood. Here we show that CAP350 and FOP (FGFR1 oncogene partner) form a centrosomal complex required for MT anchoring. We show that the C terminal domain of CAP350 interacts directly with FOP and that both proteins localize to the centrosome throughout the cell cycle. FOP also binds to EB1 and is required for localizing EB1 to the centrosome. Depletion of either CAP350, FOP, or EB1 by siRNA causes loss of MT anchoring and profound disorganization of the MT network. These results have implications for the mechanisms underlying MT anchoring at the centrosome and they attribute a key MT anchoring function to two novel centrosomal proteins, CAP350 and FOP. PMID- 16314389 TI - Interaction of Hsp90 with ribosomal proteins protects from ubiquitination and proteasome-dependent degradation. AB - Heat-shock protein 90 (Hsp90) is a molecular chaperone that plays a key role in the conformational maturation of various transcription factors and protein kinases in signal transduction. Multifunctional ribosomal protein S3 (rpS3), a component of the ribosomal small subunit, is involved in DNA repair and apoptosis. Our data show that Hsp90 binds directly to rpS3 and the functional consequence of Hsp90-rpS3 interaction results in the prevention of the ubiquitination and the proteasome-dependent degradation of rpS3, subsequently retaining the function and the biogenesis of the ribosome. Interference of Hsp90 activity by Hsp90 inhibitors appears to dissociate rpS3 from Hsp90, associate the protein with Hsp70, and induce the degradation of free forms of rpS3. Furthermore, ribosomal protein S6 (rpS6) also interacted with Hsp90 and exhibited a similar effect upon treatment with Hsp90 inhibitors. Therefore, we conclude that Hsp90 regulates the function of ribosomes by maintaining the stability of 40S ribosomal proteins such as rpS3 and rpS6. PMID- 16314390 TI - Extracellular signal-regulated kinase regulates clathrin-independent endosomal trafficking. AB - Extracellular signal-regulated kinase (Erk) is widely recognized for its central role in cell proliferation and motility. Although previous work has shown that Erk is localized at endosomal compartments, no role for Erk in regulating endosomal trafficking has been demonstrated. Here, we report that Erk signaling regulates trafficking through the clathrin-independent, ADP-ribosylation factor 6 (Arf6) GTPase-regulated endosomal pathway. Inactivation of Erk induced by a variety of methods leads to a dramatic expansion of the Arf6 endosomal recycling compartment, and intracellular accumulation of cargo, such as class I major histocompatibility complex, within the expanded endosome. Treatment of cells with the mitogen-activated protein kinase kinase (MEK) inhibitor U0126 reduces surface expression of MHCI without affecting its rate of endocytosis, suggesting that inactivation of Erk perturbs recycling. Furthermore, under conditions where Erk activity is inhibited, a large cohort of Erk, MEK, and the Erk scaffold kinase suppressor of Ras 1 accumulates at the Arf6 recycling compartment. The requirement for Erk was highly specific for this endocytic pathway, because its inhibition had no effect on trafficking of cargo of the classical clathrin dependent pathway. These studies reveal a previously unappreciated link of Erk signaling to organelle dynamics and endosomal trafficking. PMID- 16314391 TI - Biogenesis of tubular ER-to-Golgi transport intermediates. AB - Tubular transport intermediates (TTIs) have been described as one class of transport carriers in endoplasmic reticulum (ER)-to-Golgi transport. In contrast to vesicle budding and fusion, little is known about the molecular regulation of TTI synthesis, transport and fusion with target membranes. Here we have used in vivo imaging of various kinds of GFP-tagged proteins to start to address these questions. We demonstrate that under steady-state conditions TTIs represent approximately 20% of all moving transport carriers. They increase in number and length when more transport cargo becomes available at the donor membrane, which we induced by either temperature-related transport blocks or increased expression of the respective GFP-tagged transport markers. The formation and motility of TTIs is strongly dependent on the presence of intact microtubules. Microinjection of GTPgammaS increases the frequency of TTI synthesis and the length of these carriers. When Rab proteins are removed from membranes by microinjection of recombinant Rab-GDI, the synthesis of TTIs is completely blocked. Microinjection of the cytoplasmic tails of the p23 and p24 membrane proteins also abolishes formation of p24-containing TTIs. Our data suggest that TTIs are ER-to-Golgi transport intermediates that form preferentially when transport-competent cargo exists in excess at the donor membrane. We propose a model where the interaction of the cytoplasmic tails of membrane proteins with microtubules are key determinants for TTI synthesis and may also serve as a so far unappreciated model for aspects of transport carrier formation. PMID- 16314392 TI - Seventeen a-subunit isoforms of paramecium V-ATPase provide high specialization in localization and function. AB - In the Paramecium tetraurelia genome, 17 genes encoding the 100-kDa-subunit (a subunit) of the vacuolar-proton-ATPase were identified, representing by far the largest number of a-subunit genes encountered in any organism investigated so far. They group into nine clusters, eight pairs with >82% amino acid identity and one single gene. Green fluorescent protein-tagging of representatives of the nine clusters revealed highly specific targeting to at least seven different compartments, among them dense core secretory vesicles (trichocysts), the contractile vacuole complex, and phagosomes. RNA interference for two pairs confirmed their functional specialization in their target compartments: silencing of the trichocyst-specific form affected this secretory pathway, whereas silencing of the contractile vacuole complex-specific form altered organelle structure and functioning. The construction of chimeras between selected a subunits surprisingly revealed the targeting signal to be located in the C terminus of the protein, in contrast with the N-terminal targeting signal of the a-subunit in yeast. Interestingly, some chimeras provoked deleterious effects, locally in their target compartment, or remotely, in the compartment whose specific a-subunit N terminus was used in the chimera. PMID- 16314393 TI - Nuclear envelope breakdown is coordinated by both Nup358/RanBP2 and Nup153, two nucleoporins with zinc finger modules. AB - When higher eukaryotic cells transition into mitosis, the nuclear envelope, nuclear pore complexes, and nuclear lamina are coordinately disassembled. The COPI coatomer complex, which plays a major role in membrane remodeling at the Golgi, has been implicated in the process of nuclear envelope breakdown and requires interactions at the nuclear pore complex for recruitment to this new site of action at mitosis. Nup153, a resident of the nuclear pore basket, was found to be involved in COPI recruitment, but the molecular nature of the interface between COPI and the nuclear pore has not been fully elucidated. To better understand what occurs at the nuclear pore at this juncture, we have probed the role of the nucleoporin Nup358/RanBP2. Nup358 contains a repetitive zinc finger domain with overall organization similar to a region within Nup153 that is critical to COPI association, yet inspection of these two zinc finger domains reveals features that also clearly distinguish them. Here, we found that the Nup358 zinc finger domain, but not a zinc finger domain from an unrelated protein, binds to COPI and dominantly inhibits progression of nuclear envelope breakdown in an assay that robustly recapitulates this process in vitro. Moreover, the Nup358 zinc finger domain interferes with COPI recruitment to the nuclear rim. Consistent with a role for this pore protein in coordinating nuclear envelope breakdown, Nup358-specific antibodies impair nuclear disassembly. Significantly, targeting either Nup153 or Nup358 for inhibition perturbs nuclear envelope breakdown, supporting a model in which these nucleoporins play nonredundant roles, perhaps contributing to COPI recruitment platforms on both the nuclear and cytoplasmic faces of the pore. We found that an individual zinc finger is the minimal interface for COPI association, although tandem zinc fingers are optimal. These results provide new information about the critical components of nuclear membrane remodeling and lay the foundation for a better understanding of how this process is regulated. PMID- 16314394 TI - SNAP25, but not syntaxin 1A, recycles via an ARF6-regulated pathway in neuroendocrine cells. AB - Soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor (SNARE) proteins mediate cellular membrane fusion events and provide a level of specificity to donor-acceptor membrane interactions. However, the trafficking pathways by which individual SNARE proteins are targeted to specific membrane compartments are not well understood. In neuroendocrine cells, synaptosome associated protein of 25 kDa (SNAP25) is localized to the plasma membrane where it functions in regulated secretory vesicle exocytosis, but it is also found on intracellular membranes. We identified a dynamic recycling pathway for SNAP25 in PC12 cells through which plasma membrane SNAP25 recycles in approximately 3 h. Approximately 20% of the SNAP25 resides in a perinuclear recycling endosome-trans Golgi network (TGN) compartment from which it recycles back to the plasma membrane. SNAP25 internalization occurs by constitutive, dynamin-independent endocytosis that is distinct from the dynamin-dependent endocytosis that retrieves secretory vesicle constituents after exocytosis. Endocytosis of SNAP25 is regulated by ADP-ribosylation factor (ARF)6 (through phosphatidylinositol bisphosphate synthesis) and is dependent upon F-actin. SNAP25 endosomes, which exclude the plasma membrane SNARE syntaxin 1A, merge with those derived from clathrin-dependent endocytosis containing endosomal syntaxin 13. Our results characterize a robust ARF6-dependent internalization mechanism that maintains an intracellular pool of SNAP25, which is compatible with possible intracellular roles for SNAP25 in neuroendocrine cells. PMID- 16314395 TI - The role of Cdc55 in the spindle checkpoint is through regulation of mitotic exit in Saccharomyces cerevisiae. AB - Cdc55, a B-type regulatory subunit of protein phosphatase 2A, has been implicated in mitotic spindle checkpoint activity and maintenance of sister chromatid cohesion during metaphase. The spindle checkpoint is composed of two independent pathways, one leading to inhibition of the metaphase-to-anaphase transition by checkpoint proteins, including Mad2, and the other to inhibition of mitotic exit by Bub2. We show that Cdc55 is a negative regulator of mitotic exit. A cdc55 mutant, like a bub2 mutant, prematurely releases Cdc14 phosphatase from the nucleolus during spindle checkpoint activation, and premature exit from mitosis indirectly leads to loss of sister chromatid cohesion and inviability in nocodazole. The role of Cdc55 is separable from Bub2 and inhibits release of Cdc14 through a mechanism independent of the known negative regulators of mitotic exit. Epistasis experiments indicate Cdc55 acts either downstream or independent of the mitotic exit network kinase Cdc15. Interestingly, the B-type cyclin Clb2 is partially stable during premature activation of mitotic exit in a cdc55 mutant, indicating mitotic exit is incomplete. PMID- 16314396 TI - Golgi inheritance in mammalian cells is mediated through endoplasmic reticulum export activities. AB - Golgi inheritance during mammalian cell division occurs through the disassembly, partitioning, and reassembly of Golgi membranes. The mechanisms responsible for these processes are poorly understood. To address these mechanisms, we have examined the identity and dynamics of Golgi proteins within mitotic membranes using live cell imaging and electron microscopy techniques. Mitotic Golgi fragments, seen in prometaphase and telophase, were found to localize adjacent to endoplasmic reticulum (ER) export domains, and resident Golgi transmembrane proteins cycled rapidly into and out of these fragments. Golgi proteins within mitotic Golgi haze-seen during metaphase-were found to redistribute with ER markers into fragments when the ER was fragmented by ionomycin treatment. The temperature-sensitive misfolding mutant ts045VSVG protein, when localized to the Golgi at the start of mitosis, became trapped in the ER at the end of mitosis in cells shifted to 40 degrees C. Finally, reporters for Arf1 and Sar1 activity revealed that Arf1 and Sar1 undergo sequential inactivation during mitotic Golgi breakdown and sequential reactivation upon Golgi reassembly at the end of mitosis. Together, these findings support a model of mitotic Golgi inheritance that involves inhibition and subsequent reactivation of cellular activities controlling the cycling of Golgi components into and out of the ER. PMID- 16314397 TI - Mutations in tap uncouple RNA export activity from translocation through the nuclear pore complex. AB - Interactions between transport receptors and phenylalanine-glycine (FG) repeats on nucleoporins drive the translocation of receptor-cargo complexes through nuclear pores. Tap, a transport receptor that mediates nuclear export of cellular mRNAs, contains a UBA-like and NTF2-like folds that can associate directly with FG repeats. In addition, two nuclear export sequences (NESs) within the NTF2-like region can also interact with nucleoporins. The Tap-RNA complex was shown to bind to three nucleoporins, Nup98, p62, and RanBP2, and these interactions were enhanced by Nxt1. Mutations in the Tap-UBA region abolished interactions with all three nucleoporins, whereas the effect of point mutations within the NTF2-like domain of Tap known to disrupt Nxt1 binding or nucleoporin binding were nucleoporin dependent. A mutation in any of these Tap domains was sufficient to reduce RNA export but was not sufficient to disrupt Tap interaction with the NPC in vivo or its nucleocytoplasmic shuttling. However, shuttling activity was reduced or abolished by combined mutations within the UBA and either the Nxt1 binding domain or NESs. These data suggest that Tap requires both the UBA- and NTF2-like domains to mediate the export of RNA cargo, but can move through the pores independently of these domains when free of RNA cargo. PMID- 16314398 TI - Molecular classification of green tea catechin-sensitive and green tea catechin resistant prostate cancer in the TRAMP mice model by quantitative real-time PCR gene profiling. AB - We previously found that human prostate cancer (CaP) progression is accompanied by differential expression of a panel of 8 informative genes, some of which are metabolically related. Gene profiling focused on this 8-gene pack by northern blot analysis in combination with standard clinical information provided reliable prognostic prediction of human CaP. For a better insight into the potential of this 8-gene signature in tumor detection/classification and therapeutic response, we determined, by qPCR, the expression of these informative genes in the TRAMP mice model of CaP progression. The 8-genes signature resulted effective in discriminating, by linear discriminant analysis (LDA), the prostate of wild type mice from transgenic TRAMP mice developing CaP (P < 0.0002). Since it is known that Green Tea Catechin (GTC) administration to TRAMP mice results in a substantial delay of CaP progression in 80% of the animals, while 20% remain unresponsive, we determined the 8-gene signature in the prostates of GTC sensitive and GTC-resistant mice. LDA discriminated benign tissue from CaP (i.e. wild-type + chemoprevented, GTC-sensitive TRAMP mice, in which CaP progression was delayed, was discriminated from TRAMP mice + GTC-resistant TRAMP mice, in which CaP developed irrespective of GTC administration; P < 0.01). Moreover, GTC sensitive TRAMP mice bearing CaP were discriminated from GTCs-resistant ones, (P = 0.0001). These results show that qPCR gene profiling, based on the signature of the 8-genes selected by us, could represent an appropriate means for studying the biological behavior of CaP, which may lead to identifying new tools of potential prognostic value, in that a molecular classification for the presence/absence of cancer and for discriminating GTCs-responsive from GTC-resistant CaP is provided. PMID- 16314399 TI - Joint effects of single nucleotide polymorphisms in P53BP1 and p53 on breast cancer risk in a Chinese population. AB - p53-binding protein 1 (P53BP1), a central transducer of DNA-damage signals to p53, is required for both intra-S-phase and G2-M checkpoints, suggesting that these two proteins may work together in the p53-mediated transcriptional activation and DNA damage-repair signaling pathways. Because the p53-binding region of 53BP1 maps to the C-terminal BRCT domains, which are homologous to those found in the breast cancer protein BRCA1, we hypothesized that genetic variation in P53BP1 and p53 may contribute to breast cancer predisposition. To test this hypothesis, we simultaneously genotyped single nucleotide polymorphisms of T-885G, Glu353Asp, and Gln1136Lys in P53BP1 and Arg72Pro in p53 in a case control study of 404 breast cancer cases and 472 cancer-free controls. We found that the P53BP1 variant genotypes (alleles) of T-885G and Gln1136Lys were associated with a significantly increased risk of breast cancer among p53 Pro/Pro carriers (OR=2.36, 95% CI 1.16-4.83 for -885TG/GG; OR=2.24, 95% CI 1.15-4.37 for 1136Gln/Lys+Lys/Lys and OR=2.82, 95% CI 1.15-6.94 for >4 variant alleles of these 3 loci). In addition, the variant genotypes of above 3 loci of P53BP1 were significantly associated with elevated risk of progesterone receptor (PR) negative breast cancer, and the T-885G and Gln1136Lys with estrogen receptor (ER) negative breast cancer. Furthermore, we found a significant gene-gene interaction between P53BP1 Gln1136Lys and p53 Arg72Pro variants in relation to breast cancer, and the OR of interaction for the presence of both P53BP1 1136Gln/Lys+Lys/Lys and p53 72Arg/Pro+Pro/Pro genotypes was 1.93 (95% CI 1.06-3.52) (P=0.031 for interaction). These findings indicate that the SNPs in P53BP1 and p53 jointly contribute to breast cancer risk, particularly ER (-) or PR (-) breast cancer, and the p53 Arg72Pro polymorphism may serve as a risk modifier. Further functional studies are needed to confirm our findings. PMID- 16314401 TI - Prevention of chemotherapy- and radiotherapy-induced emesis: results of the 2004 Perugia International Antiemetic Consensus Conference. AB - BACKGROUND: In the late 1990s, several professional organizations convened antiemetic guideline groups and published the findings of these expert panels. Each of these documents was based on analyses of the available published trials and provided nearly similar recommendations. Nonetheless, small differences in emetic risk categories and treatment recommendations led to confusion in antiemetics selection. With the emergence of new findings and agents since the guidelines were initially published, many of the oncology professional societies have updated the antiemetic guidelines. MATERIALS AND METHODS: A literature review up to March 2004 was carried out using MEDLINE with evaluation of the evidence by an expert panel composed of 23 oncology professionals in clinical medicine, medical oncology, radiation oncology, oncology nursing, statistics, pharmacy, medical policy and decision making, and pharmacology. The experts represented nine oncology professional societies and came from 11 different countries on four continents. RESULTS: Recommendations on antiemetic regimens to prevent emesis induced by high, moderate, low and minimal risk chemotherapy were suggested as well as management of anticipatory emesis. Furthermore, recommendations for refractory emesis, emesis induced by high-dose chemotherapy and radiotherapy and for antiemetics in children receiving chemotherapy were elaborated. CONCLUSIONS: Recommendations about antiemetic prophylaxis in patients receiving treatment with chemo- and radiotherapy have been updated by representatives of nine oncological organizations. PMID- 16314402 TI - Comparison of in vitro assays of cellular toxicity in the human hepatic cell line HepG2. AB - Cytotoxicity testing allows determining whether a compound or extract contains significant quantities of biologically harmful chemicals. Cytotoxicity test methods are useful for screening because they serve to separate toxic from nontoxic materials, providing predictive evidence of compound safety. However, a wide range of assays measuring different aspects of cell death is available in the market, but it is difficult to determine which one(s) to use when evaluating a selection of compounds. The objective of this study was to compare different commercially available in vitro assays for cytotoxicity in HepG2 cells according to its sensitivity, reproducibility, simplicity, cost, and speed. The assays evaluated included Alamar Blue for the measurement of mitochondrial activity, ATPlite and ViaLight for the determination of cellular adenosine triphosphate (ATP), ToxiLight as an indicator of cellular necrosis, and Caspase-3 Fluorometric Assay, Apo-ONE Caspase-3/7 Homogeneous Assay, and Caspase-Glo for the determination of caspase-3/7 activity. All assays were performed using 4 compounds of previously reported cytotoxic activity: DMSO, butyric acid, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), and camptothecine. Overall, it was concluded that the best way to evaluate the potential cytotoxicity of a compound is to employ a battery of assays that focus on different aspects of cell death. In this case, the focus has been on ATP levels, cell necrosis, and capsase 3/7 activation. Many other kits are commercially available in the market for these and other aspects of necrosis and/or apoptosis. However, the use of ViaLight Plus, ToxiLight, and Caspase-3 Fluorometric Assay resulted in the most useful combination when working with HepG2 cells. PMID- 16314403 TI - Development of a novel dual CCR5-dependent and CXCR4-dependent cell-cell fusion assay system with inducible gp160 expression. AB - In the current study, a novel coreceptor-specific cell-cell fusion (CCF) assay system is reported. The system possesses the following features: dual CCR5 dependent and CXCR4-dependent CCF assays, all stable cell lines, inducible expression of gp160 to minimize cytotoxicity, robust luciferase reporter, and 384 well format. These assays have been validated using various known HIV entry inhibitors targeting various stages of the HIV entry/fusion process, including fusion inhibitors, gp120 inhibitors, CCR5 antagonists, CCR5 antibodies, and CXCR4 antagonists. IC50 data generated from this assay system were well correlated to that from the antiviral assays. The effects of DMSO on this assay system were assessed, and a 2- to 3-fold increase in luciferase activity was observed in the presence of 0.05% to 2% DMSO. Although cell-cell fusion efficiency was enhanced, no changes in drug response kinetics for entry inhibitors were found in the presence of 0.1% or 0.5% DMSO. This assay system has been successfully used for the identification and characterization of thousands of CCR5 inhibitors. PMID- 16314404 TI - Identification of novel Kv1.3 blockers using a fluorescent cell-based ion channel assay. AB - A functional cell-based assay was developed using a generic proprietary assay protocol, based on a membrane-potential sensitive dye, for the identification of small-molecule antagonists against the Kv1.3 potassium ion channel. A high throughput screen (HTS) was subsequently performed with 20,000 compounds from the Evotec library, preselected using known small molecule antagonists for both sodium and potassium ion channels. Following data analysis, the hit rate was measured at 1.72%, and subsequent dose-response analysis of selected hits showed a high hit confirmation rate yielding approximately 50 compounds with an apparent IC50 value lower than 10 microM. Subsequent electrophysiological characterization of selected hits confirmed the initial activity and potency of the identified hits on the Kv1.3 target and also selectivity toward Kv1.3 through measurements on HERG as well as Kv1.3-expressing cell lines. Follow-up structure-activity relationship analysis revealed a variety of different clusters distributed throughout the library as well as several singlicates. In comparison to known Kv1.3 blockers, new chemical entities and scaffolds showing potency and selectivity against the Kv1.3 ion channel were detected. In addition, a screening strategy for ion channel drug discovery HTS, medicinal chemistry, and electrophysiology is presented. PMID- 16314405 TI - A radioligand binding assay for antitubulin activity in tumor cells. AB - The benzamide RH-5854 is shown to be highly potent toward tumor cells and to arrest nuclear division by a highly specific covalent binding to the beta-subunit of tubulin in the colchicine binding region. Binding of 3H-RH-5854 to beta tubulin in HCT-116 colon cancer cells is saturable and has been exploited in the development of a cell-based competitive binding assay, which allows antitubulin effects to be detected in whole cells. 3H-RH-5854 binding is strongly inhibited by preincubating the cells with compounds that bind to the colchicine site and with paclitaxel. Binding of 3H-RH-5854 is enhanced by preincubating the cells with vinblastine but not by other agents that bind at or near the vinblastine site (ansamitocin P-3 and phomopsin A). Various cytotoxic agents that do not act on tubulin do not affect binding of 3H-RH-5854 in HCT-116 cells, demonstrating specificity of the assay for detection of antitubulin activity. As an alternative to traditional assays that employ isolated brain tubulin, the 3HRH-5854 binding assay enables screening for antitubulin effects directly in tumor cells, providing an assay that accounts for cell-specific criteria that influence sensitivity such as different tubulin isotypes, tubulin mutations, drug metabolism, and efflux mechanisms. PMID- 16314406 TI - Microarrays for the functional analysis of the chemical-kinase interactome. AB - A central challenge in chemical biology is profiling the activity of a large number of chemical structures against hundreds of biological targets, such as kinases. Conventional 32P-incorporation or immunoassay of phosphorylated residues produces high-quality signals for monitoring kinase reactions but is difficult to use in high-throughput screening (HTS) because of cost and the need for well plate washing. The authors report a method for densely archiving compounds in nanodroplets on peptide or protein substrate-coated microarrays for subsequent profiling by aerosol deposition of kinases. Each microarray contains over 6000 reaction centers (1.0 nL each) whose phosphorylation progress can be detected by immunofluorescence. For p60c-src, the microarray produced a signal-to-background ratio of 36.3 and Z' factor of 0.63 for HTS and accurate enzyme kinetic parameters (KmATP = 3.3 microM) and IC50 values for staurosporine (210 nM) and PP2 (326 nM) at 10 microM adenosine triphosphate (ATP). Similarly, B-Raf phosphorylation of MEK-coated microarrays was inhibited in the nanoliter reactions by GW5074 at the expected IC50 of 9 nM. Common kinase inhibitors were printed on microarrays, and their inhibitory activities were systematically profiled against B-Raf (V599E), KDR, Met, Flt-3 (D835Y), Lyn, EGFR, PDGFRbeta, and Tie2. All results indicate that this platform is well suited for kinetic analysis, HTS, large-scale IC50 determinations, and selectivity profiling. PMID- 16314407 TI - Rapid screening of blood-brain barrier penetration of drugs using the immobilized artificial membrane phosphatidylcholine column chromatography. AB - The chromatographic capacity factors (kIAM) of 23 structurally diverse drugs were measured by the immobilized artificial membrane (IAM) phosphatidylcholine chromatography for the prediction of blood-brain barrier (BBB) penetration. The kIAM was determined using the mobile phase consisting of acetonitrile:DPBS (20:80 v/v) and corrected for the molar volume of the solutes (kIAM/MWn). The correlation between kIAM/MWn and CNS penetration was highest when measured at pH 5.5 with the power function of n = 4. This in vitro prediction method was validated with 7 newly synthesized PDE-4 inhibitors. The relationship between in vivo plasma-to-brain concentration ratios and in vitro CNS penetration was excellent (r = 0.959). The developed in vitro prediction method may be used as a rapid screening tool for BBB penetration of drugs with passive transport mechanism, with high success, low cost, and reproducibility. PMID- 16314408 TI - Biomarker discovery, disease classification, and similarity query processing on high-throughput MS/MS data of inborn errors of metabolism. AB - In newborn errors of metabolism, biomarkers are urgently needed for disease screening, diagnosis, and monitoring of therapeutic interventions. This article describes a 2-step approach to discover metabolic markers, which involves (1) the identification of marker candidates and (2) the prioritization of them based on expert knowledge of disease metabolism. For step 1, the authors developed a new algorithm, the biomarker identifier (BMI), to identify markers from quantified diseased versus normal tandem mass spectrometry data sets. BMI produces a ranked list of marker candidates and discards irrelevant metabolites based on a quality measure, taking into account the discriminatory performance, discriminatory space, and variance of metabolites' concentrations at the state of disease. To determine the ability of identified markers to classify subjects, the authors compared the discriminatory performance of several machine-learning paradigms and described a retrieval technique that searches and classifies abnormal metabolic profiles from a screening database. Seven inborn errors of metabolism-- phenylketonuria (PKU), glutaric acidemia type I (GA-I), 3 methylcrotonylglycinemia deficiency (3-MCCD), methylmalonic acidemia (MMA), propionic acidemia (PA), medium-chain acylCoAdehydrogenase deficiency (MCADD), and 3-OH long-chain acyl CoA dehydrogenase deficiency (LCHADD)-were investigated. All primarily prioritized marker candidates could be confirmed by literature. Some novel secondary candidates were identified (i.e., C16:1 and C4DC for PKU, C4DC for GA-I, and C18:1 forMCADD), which require further validation to confirm their biochemical role during health and disease. PMID- 16314409 TI - Xyloglucan is recognized by carbohydrate-binding modules that interact with beta glucan chains. AB - Enzyme systems that attack the plant cell wall contain noncatalytic carbohydrate binding modules (CBMs) that mediate attachment to this composite structure and play a pivotal role in maximizing the hydrolytic process. Although xyloglucan, which includes a backbone of beta-1,4-glucan decorated primarily with xylose residues, is a key component of the plant cell wall, CBMs that bind to this polymer have not been identified. Here we showed that the C-terminal domain of the modular Clostridium thermocellum enzyme CtCel9D-Cel44A (formerly known as CelJ) comprises a novel CBM (designated CBM44) that binds with equal affinity to cellulose and xyloglucan. We also showed that accommodation of xyloglucan side chains is a general feature of CBMs that bind to single cellulose chains. The crystal structures of CBM44 and the other CBM (CBM30) in CtCel9D-Cel44A display a beta-sandwich fold. The concave face of both CBMs contains a hydrophobic platform comprising three tryptophan residues that can accommodate up to five glucose residues. The orientation of these aromatic residues is such that the bound ligand would adopt the twisted conformation displayed by cello-oligosaccharides in solution. Mutagenesis studies confirmed that the hydrophobic platform located on the concave face of both CBMs mediates ligand recognition. In contrast to other CBMs that bind to single polysaccharide chains, the polar residues in the binding cleft of CBM44 play only a minor role in ligand recognition. The mechanism by which these proteins are able to recognize linear and decorated beta 1,4-glucans is discussed based on the structures of CBM44 and the other CBMs that bind single cellulose chains. PMID- 16314410 TI - Structural and mutagenesis studies on the cytochrome c peroxidase from Rhodobacter capsulatus provide new insights into structure-function relationships of bacterial di-heme peroxidases. AB - Cytochrome c peroxidases (CCP) play a key role in cellular detoxification by catalyzing the reduction of hydrogen peroxide to water. The di-heme CCP from Rhodobacter capsulatus is the fastest enzyme (1060 s(-1)), when tested with its physiological cytochrome c substrate, among all di-heme CCPs characterized to date and has, therefore, been an attractive target to investigate structure function relationships for this family of enzymes. Here, we combine for the first time structural studies with site-directed mutagenesis and spectroscopic studies of the mutant enzymes to investigate the roles of amino acid residues that have previously been suggested to be important for activity. The crystal structure of R. capsulatus at 2.7 Angstroms in the fully oxidized state confirms the overall molecular scaffold seen in other di-heme CCPs but further reveals that a segment of about 10 amino acids near the peroxide binding site is disordered in all four molecules in the asymmetric unit of the crystal. Structural and sequence comparisons with other structurally characterized CCPs suggest that flexibility in this part of the molecular scaffold is an inherent molecular property of the R. capsulatus CCP and of CCPs in general and that it correlates with the levels of activity seen in CCPs characterized, thus, far. Mutagenesis studies support the spin switch model and the roles that Met-118, Glu-117, and Trp-97 play in this model. Our results help to clarify a number of aspects of the debate on structure-function relationships in this family of bacterial CCPs and set the stage for future studies. PMID- 16314411 TI - E6AP and calmodulin reciprocally regulate estrogen receptor stability. AB - Estrogen promotes the proliferation of human breast epithelial cells by interacting with the estrogen receptor (ER). Physiological responses of cells to estrogen are regulated in part by degradation of the ER. Previous studies revealed that calmodulin binds directly to the ER, thereby enhancing its stability. Consistent with these findings, cell-permeable calmodulin antagonists dramatically reduced the number of ER in MCF-7 human breast epithelial cells. Here we investigated the molecular mechanism by which calmodulin attenuates ER degradation. MG132 and lactacystin, inhibitors of the ubiquitin-proteasome pathway, prevented the calmodulin antagonist CGS9343B from reducing the amount of ER in MCF-7 cells. In contrast, protease inhibitors afforded no protection. Moreover, CGS9343B enhanced ER ubiquitination. A point mutant ER construct that is unable to bind calmodulin, termed ERDeltaCaM, is ubiquitinated to a greater extent than wild type ER. The ubiquitin-protein isopeptide ligase E6-associated protein (E6AP) associated with and promoted the degradation of ER. The possible convergence of calmodulin and E6AP on ER degradation was examined. ERDeltaCaM bound E6AP with higher affinity than that of wild type ER. Moreover, calmodulin attenuated the in vitro interaction between ER and E6AP in a Ca(2+)-dependent manner. Collectively, our data reveal that E6AP is a component of ER degradation via the ubiquitin-proteasome pathway and that Ca(2+)/calmodulin modulates this degradation mechanism. These results have potential implications for the development of selectively targeted therapeutic agents for breast cancer. PMID- 16314412 TI - Paratope determination of the antithrombotic antibody 82D6A3 based on the crystal structure of its complex with the von Willebrand factor A3-domain. AB - The antithrombotic monoclonal antibody 82D6A3 is directed against amino acids Arg 963, Pro-981, Asp-1009, Arg-1016, Ser-1020, Met-1022, and His-1023 of the von Willebrand factor A3-domain (Vanhoorelbeke, K., Depraetere, H., Romijn, R. A., Huizinga, E., De Maeyer, M., and Deckmyn, H. (2003) J. Biol. Chem. 278, 37815 37821). By this, it potently inhibits the interaction of von Willebrand factor to collagens, which is a prerequisite for blood platelet adhesion to the injured vessel wall at sites of high shear. To fully understand the mode of action of 82D6A3 at the molecular level, we resolved its crystal structure in complex with the A3-domain and fine mapped its paratope by construction and characterization of 13 mutants. The paratope predominantly consists of two short sequences in the heavy chain CDR1 (Asn-31 and Tyr-32) and CDR3 (Asp-99, Pro-101, Tyr-102 and Tyr 103), forming one patch on the surface of the antibody. Trp-50 of the heavy and His-49 of the light chain, both situated adjacent to the patch, play ancillary roles in antigen binding. The crystal structure furthermore confirms the epitope location, which largely overlaps with the collagen binding site deduced from mutagenesis of the A3-domain (Romijn, R. A., Westein, E., Bouma, B., Schiphorst, M. E., Sixma, J. J., Lenting, P. J., and Huizinga, E. G. (2003) J. Biol. Chem. 278, 15035-15039). We herewith further consolidate the location of the collagen binding site and reveal that the potent action of the antibody is due to direct competition for the same interaction site. This information allows the design of a paratope-mimicking peptide with antithrombotic properties. PMID- 16314413 TI - Arrestin-mediated ERK activation by gonadotropin-releasing hormone receptors: receptor-specific activation mechanisms and compartmentalization. AB - Activation of seven-transmembrane region receptors typically causes their phosphorylation with consequent arrestin binding and desensitization. Arrestins also act as scaffolds, mediating signaling to Raf and ERK and, for some receptors, inhibiting nuclear translocation of ERK. GnRH receptors (GnRHRs) act via Gq/11 to stimulate the phospholipase C/Ca2+/protein kinase C (PKC) cascade and the Raf/MEK/ERK cassette. Uniquely, type I mammalian GnRHRs lack the C-tails that are found in other seven-transmembrane region receptors (including nonmammalian GnRHRs) and are implicated in arrestin binding. Here we have compared ERK signaling by human GnRHRs (hGnRHRs) and Xenopus GnRHRs (XGnRHRs). In HeLa cells, XGnRHRs underwent rapid and arrestin-dependent internalization and caused arrestin/green fluorescent protein (GFP) translocation to the membrane and endosomes, whereas hGnRHRs did not. Internalized XGnRHRs were co-localized with arrestin-GFP, whereas hGnRHRs were not. Both receptors mediated transient ERK phosphorylation and nuclear translocation (revealed by immunohistochemistry or by imaging of co-transfected ERK2-GFP), and for both, ERK phosphorylation was reduced by PKC inhibition but not by inhibiting epidermal growth factor receptor autophosphorylation. In the presence of PKC inhibitor, Deltaarrestin-(319-418) blocked XGnRHR-mediated, but not hGnRHR-mediated, ERK phosphorylation. When receptor number was varied, hGnRHRs activated phospholipase C and ERK more efficiently than XGnRHRs but were less efficient at causing ERK2-GFP translocation. At high receptor number, XGnRHRs and hGnRHRs both caused ERK2-GFP translocation to the nucleus, but at low receptor number, XGnRHRs caused ERK2-GFP translocation, whereas hGnRHRs did not. Thus, experiments with XGnRHRs have revealed the first direct evidence of arrestin-mediated (probably G protein independent) GnRHR signaling, whereas those with hGnRHRs imply that scaffolds other than arrestins can determine GnRHR effects on ERK compartmentalization. PMID- 16314414 TI - Hoxd13 and Hoxa13 directly control the expression of the EphA7 Ephrin tyrosine kinase receptor in developing limbs. AB - Hoxa and Hoxd genes, related to the Drosophila Abd-B gene, display regionally restricted expression patterns and are necessary for the formation of the limb skeletal elements. Hox genes encode transcription factors, which are supposed to control the expression of a series of downstream target genes, whose nature has remained largely elusive. Several genes were identified that are differentially expressed in relation to Hox gene activity; few studies, however, explored their direct regulation by Hox proteins. Ephrin tyrosine kinase receptors and ephrins have been proposed as Hox targets, and recently, evidence was gained for their role in limb development. The expression of the EphA7 gene in developing limbs was shown to correlate with the expression of Hoxa13 and Hoxd13; however, its direct regulation by these genes has never been assessed. We have characterized the EphA7 promoter region and show that it contains multiple binding sites for paralog group 13 Hox proteins. We found that one of these sites is bound in vivo by HOXA13 and HOXD13 and by endogenous Hoxd13 in developing mouse limbs. Moreover, we show that HOXD13 and HOXA13 activate transcription from the EphA7 promoter and that a mutation of the HOXA13/HOXD13 binding site was sufficient to abolish activation. Conversely, the HOXD13(147L) mutation, identified in patients displaying a novel brachydactyly-polydactyly syndrome, does not bind to in vivo, and fails to transactivate the EphA7 promoter. These results establish that EphA7 is a direct downstream target of Hoxd13 and Hoxa13 during limb development, thus providing further insight into the regulatory networks that control limb patterning. PMID- 16314415 TI - Annealing prion protein amyloid fibrils at high temperature results in extension of a proteinase K-resistant core. AB - Amyloids are highly ordered, rigid beta-sheet-rich structures that appear to have minimal dynamic flexibility in individual polypeptide chains. Here, we demonstrate that substantial conformational rearrangements occur within mature amyloid fibrils produced from full-length mammalian prion protein. The rearrangement results in a substantial extension of a proteinase K-resistant core and is accompanied by an increase in the beta-sheet-rich conformation. The conformational rearrangement was induced in the presence of low concentrations of Triton X-100 either by brief exposure to 80 degrees C or, with less efficacy, by prolonged incubation at 37 degrees C at pH 7.5 and is referred to here as "annealing." Upon annealing, amyloid fibrils acquired a proteinase K-resistant core identical to that found in bovine spongiform encephalopathy-specific scrapie associated prion protein. Annealing was also observed when amyloid fibrils were exposed to high temperatures in the absence of detergent but in the presence of brain homogenate. These findings suggest that the amyloid fibrils exist in two conformationally distinct states that are separated by a high energy barrier and that yet unknown cellular cofactors may facilitate transition of the fibrils into thermodynamically more stable state. Our studies provide new insight into the complex behavior of prion polymerization and highlight the annealing process, a previously unknown step in the evolution of amyloid structures. PMID- 16314416 TI - AMP deamination delays muscle acidification during heavy exercise and hypoxia. AB - In silico studies carried out by using a computer model of oxidative phosphorylation and anaerobic glycolysis in skeletal muscle demonstrated that deamination of AMP to IMP during heavy short term exercise and/or hypoxia lessens the acidification of myocytes. The concerted action of adenylate kinase and AMP deaminase, leading to a decrease in the total adenine nucleotide pool, constitutes an additional process consuming ADP and producing ATP. It diminishes the amount of ADP that must be converted to ATP by other processes in order to meet the rate of ADP production by ATPases (because the adenylate kinase + AMP deaminase system produces only 1 ATP per 2 ADPs used, ATP consumption is not matched by ATP production, and the reduction of the total adenine nucleotide pool occurs mostly at the cost of [ATP]). As a result, the rate of ADP consumption by other processes may be lowered. This effect concerns mostly ADP consumption by anaerobic glycolysis that is inhibited by AMP deamination-induced decrease in [ADP] and [AMP], and not oxidative phosphorylation, because during heavy exercise and/or hypoxia [ADP] is significantly greater than the Km value of this process for ADP. The resultant reduction of proton production by anaerobic glycolysis enables us to delay the termination of exercise because of fatigue and/or to diminish cell damage. PMID- 16314417 TI - Interacting residues in an activated state of a G protein-coupled receptor. AB - Ste2p, the G protein-coupled receptor (GPCR) for the tridecapeptide pheromone alpha-factor of Saccharomyces cerevisiae, was used as a model GPCR to investigate the role of specific residues in the resting and activated states of the receptor. Using a series of biological and biochemical analyses of wild-type and site-directed mutant receptors, we identified Asn(205) as a potential interacting partner with the Tyr(266) residue. An N205H/Y266H double mutant showed pH dependent functional activity, whereas the N205H receptor was non-functional and the Y266H receptor was partially active indicating that the histidine 205 and 266 residues interact in an activated state of the receptor. The introduction of N205K or Y266D mutations into the P258L/S259L constitutively active receptor suppressed the constitutive activity; in contrast, the N205K/Y266D/P258L/S259L quadruple mutant was fully constitutively active, again indicating an interaction between residues at the 205 and 206 positions in the receptor-active state. To further test this interaction, we introduced the N205C/Y266C, F204C/Y266C, and N205C/A265C double mutations into wild-type and P258L/S259L constitutively active receptors. After trypsin digestion, we found that a disulfide-cross-linked product, with the molecular weight expected for a receptor fragment with a cross link between N205C and Y266C, formed only in the N205C/Y266C constitutively activated receptor. This study represents the first experimental demonstration of an interaction between specific residues in an active state, but not the resting state, of Ste2p. The information gained from this study should contribute to an understanding of the conformational differences between resting and active states in GPCRs. PMID- 16314418 TI - Regulation of p53 by activated protein kinase C-delta during nitric oxide-induced dopaminergic cell death. AB - Selective cell death of dopaminergic neurons in the substantia nigra is the major cause of Parkinson disease. Current evidence suggests that this cell death could be mediated by nitric oxide by-products such as nitrate and peroxynitrite. Because protein kinase C (PKC)-delta is implicated in apoptosis of various cell types, we studied its roles and activation mechanisms in nitric oxide (NO) induced apoptosis of SN4741 dopaminergic cells. When cells were treated with sodium nitroprusside (SNP), a NO donor, endogenous PKC-delta was nitrated and activated. Immunoprecipitation revealed that p53 co-immunoprecipitated with PKC delta and was phosphorylated at the 15th serine residue in SNP-treated cells. An in vitro kinase assay revealed that p53 was directly phosphorylated by SNP activated PKC-delta. The p53 Ser-15 phosphorylation was suppressed in SNP-treated cells when the NO-mediated activation of PKC-delta was inhibited by rottlerin or (-)-epigallocatechin gallate. Within 3 h of p53 phosphorylation, its protein levels increased because of decreased ubiquitin-dependent proteosomal proteolysis, whereas the protein levels of MDM2, ubiquitin-protein isopeptide ligase, were down-regulated in a p53 phosphorylation-dependent fashion. Taken together, these results demonstrate that nitration-mediated activation of PKC delta induces the phosphorylation of the Ser-15 residue in p53, which increases its protein stability, thereby contributing to the nitric oxide-mediated apoptosis-like cell death pathway. These findings may be expanded to provide new insight into the cellular mechanisms of Parkinson disease. PMID- 16314419 TI - Nitration of tyrosine 92 mediates the activation of rat microsomal glutathione s transferase by peroxynitrite. AB - There is increasing evidence that protein function can be modified by nitration of tyrosine residue(s), a reaction catalyzed by proteins with peroxidase activity, or that occurs by interaction with peroxynitrite, a highly reactive oxidant formed by the reaction of nitric oxide with superoxide. Although there are numerous reports describing loss of function after treatment of proteins with peroxynitrite, we recently demonstrated that the microsomal glutathione S transferase 1 is activated rather than inactivated by peroxynitrite and suggested that this could be attributed to nitration of tyrosine residues rather than to other effects of peroxynitrite. In this report, the nitrated tyrosine residues of peroxynitrite-treated microsomal glutathione S-transferase 1 were characterized by mass spectrometry and their functional significance determined. Of the seven tyrosine residues present in the protein, only those at positions 92 and 153 were nitrated after treatment with peroxynitrite. Three mutants (Y92F, Y153F, and Y92F, Y153F) were created using site-directed mutagenesis and expressed in LLC PK1 cells. Treatment of the microsomal fractions of these cells with peroxynitrite resulted in an approximately 2-fold increase in enzyme activity in cells expressing the wild type microsomal glutathione S-transferase 1 or the Y153F mutant, whereas the enzyme activity of Y92F and double site mutant was unaffected. These results indicate that activation of microsomal glutathione S transferase 1 by peroxynitrite is mediated by nitration of tyrosine residue 92 and represents one of the few examples in which a gain in function has been associated with nitration of a specific tyrosine residue. PMID- 16314420 TI - Lysosomal sialidase (neuraminidase-1) is targeted to the cell surface in a multiprotein complex that facilitates elastic fiber assembly. AB - We have established previously that the 67-kDa elastin-binding protein (EBP), identical to the spliced variant of beta-galactosidase, acts as a recyclable chaperone that facilitates secretion of tropoelastin. (Hinek, A., Keeley, F. W., and Callahan, J. W. (1995) Exp. Cell Res. 220, 312-324). We now demonstrate that EBP also forms a cell surface-targeted molecular complex with protective protein/cathepsin A and sialidase (neuraminidase-1), and provide evidence that this sialidase activity is a prerequisite for the subsequent release of tropoelastin. We found that treatment with sialidase inhibitors repressed assembly of elastic fibers in cultures of human skin fibroblasts, aortic smooth muscle cells, and ear cartilage chondrocytes and caused impaired elastogenesis in developing chick embryos. Fibroblasts derived from patients with congenital sialidosis (primary deficiency of neuraminidase-1) and galactosialidosis (secondary deficiency of neuraminidase-1) demonstrated impaired elastogenesis, which could be reversed after their transduction with neuraminidase-1 cDNA or after treatment with bacterial sialidase, which has a similar substrate specificity to human neuraminidase-1. We postulate that neuraminidase-1 catalyzes removal of the terminal sialic acids from carbohydrate chains of microfibrillar glycoproteins and other adjacent matrix glycoconjugates, unmasking their penultimate galactosugars. In turn, the exposed galactosugars interact with the galectin domain of EBP, thereby inducing the release of transported tropoelastin molecules and facilitating their subsequent assembly into elastic fibers. PMID- 16314421 TI - Modulation of neural carbohydrate epitope expression in Drosophila melanogaster cells. AB - Neural pathways in invertebrates are often tracked using anti-horseradish peroxidase, a cross-reaction due to the presence of core alpha1,3-fucosylated N glycans. In order to investigate the molecular basis of this epitope in a cellular context, we compared two Drosophila melanogaster cell lines: the S2 and the neuronal-like BG2-c6 cell lines. As shown by mass spectrometric and chromatographic analyses, only the BG2-c6 cell line expresses alpha1,3/alpha1,6 difucosylated N-glycans, a result that correlates with anti-horseradish peroxidase binding. Of all four alpha1,3-fucosyltransferase homologues previously identified, the core alpha1,3-fucosyltransferase (FucTA; EC 2.4.1.214) is expressed in the neuronal cell line as well as throughout fly development and in heads and bodies of flies of both sexes. This pattern is distinctive in comparison with the expression of the other three alpha1,3-fucosyltransferase homologues (FucTB, FucTC, and FucTD). Furthermore, only transfection of FucTA cDNA into S2 cells resulted in expression of the anti-horseradish peroxidase epitope, a result compatible with its substrate specificity in vitro. Finally, silencing of FucTA by RNAi in the neuronal cell line led to a significant reduction of anti-horseradish peroxidase binding. The present study, in conjunction with our previous in vitro data, thereby shows that FucTA is indispensable for expression of the neural carbohydrate epitope in Drosophila cells. PMID- 16314422 TI - G-protein-coupled receptor agonists activate endogenous phospholipase Cepsilon and phospholipase Cbeta3 in a temporally distinct manner. AB - Phospholipase Cepsilon (PLCepsilon) is one of the newest members of the phosphatidylinositol-specific phospholipase C (PLC) family. Previous studies have suggested that G-protein-coupled receptors (GPCRs) stimulate phosphoinositide (PI) hydrolysis by activating PLCbeta isoforms through G(q) family G proteins and Gbetagamma subunits. Using RNA interference to knock down PLC isoforms, we demonstrate that the GPCR agonists endothelin (ET-1), lysophosphatidic acid (LPA), and thrombin, acting through endogenous receptors, couple to both endogenous PLCepsilon and the PLCbeta isoform, PLCbeta3, in Rat-1 fibroblasts. Examination of the temporal activation of these PLC isoforms, however, reveals agonist- and isoform-specific profiles. PLCbeta3 is activated acutely within the first minute of ET-1, LPA, or thrombin stimulation but does not contribute to sustained PI hydrolysis induced by LPA or thrombin and accounts for only part of ET-1 sustained stimulation. PLCepsilon, on the other hand, predominantly accounts for sustained PI hydrolysis. Consistent with this observation, reconstitution of PLCepsilon in knockdown cells dose-dependently increases sustained, but not acute, agonist-stimulated PI hydrolysis. Furthermore, combined knockdown of both PLCepsilon and PLCbeta3 additively inhibits PI hydrolysis, suggesting independent regulation of each isoform. Importantly, ubiquitination of inositol 1,4,5 trisphosphate receptors correlates with sustained, but not acute, activation of PLCepsilon or PLCbeta3. In conclusion, GPCR agonists ET-1, LPA, and thrombin activate endogenous PLCepsilon and PLCbeta3 in Rat-1 fibroblasts. Activation of these PLC isoforms displays agonist-specific temporal profiles; however, PLCbeta3 is predominantly involved in acute and PLCepsilon in sustained PI hydrolysis. PMID- 16314423 TI - N-myc down-regulated gene 1 modulates the response of term human trophoblasts to hypoxic injury. AB - The placenta is susceptible to diverse insults during human pregnancy. The expression of the protein N-myc down-regulated gene 1 (NDRG1) is regulated during cell proliferation, differentiation, and in response to stress. Nevertheless, the function of this protein in humans remains unknown. We tested the hypothesis that NDRG1 is up-regulated in hypoxic primary human trophoblasts and that NDRG1 modulates trophoblast response to hypoxia. We initially demonstrated that the expression of NDRG1 is enhanced in primary human trophoblasts exposed to hypoxia. Importantly, we found a similar increase in NDRG1 expression in placental samples derived from either singleton gestations complicated by intrauterine growth restriction or from dizygotic twin gestation where one twin exhibited growth restriction. Having established efficient lentivirus-mediated transfection of primary human trophoblasts, we overexpressed NDRG1 in trophoblasts, which resulted in enhanced trophoblast differentiation. In contrast, lentivirus-driven short interfering RNA-mediated silencing of NDRG1 diminished trophoblast viability and differentiation. Consistent with these results, NDRG1 reduced the expression level of p53 in trophoblasts cultured in standard or hypoxic conditions. Furthermore, NDRG1 expression was regulated by the activity of SIRT1 (Sir2-like protein 1), which promotes cell survival. Together, our data indicate that NDRG1 interacts with SIRT1/p53 signaling to attenuate hypoxic injury in human trophoblasts. PMID- 16314424 TI - Ataxin-7 can export from the nucleus via a conserved exportin-dependent signal. AB - Spinocerebellar ataxia type 7 is a progressive neurodegenerative disorder caused by a CAG DNA triplet repeat expansion leading to an expanded polyglutamine tract in the ataxin-7 protein. Ataxin-7 appears to be a transcription factor and a component of the STAGA transcription coactivator complex. Here, using live cell imaging and inverted fluorescence recovery after photobleaching, we demonstrate that ataxin-7 has the ability to export from the nucleus via the CRM-1/exportin pathway and that ataxin-7 contains a classic leucine-type nuclear export signal (NES). We have precisely defined the location of this NES in ataxin-7 and found it to be fully conserved in all vertebrate species. Polyglutamine expansion was seen to reduce the nuclear export rate of mutant ataxin-7 relative to wild-type ataxin-7. Subtle point mutation of the NES in polyglutamine expanded ataxin-7 increased toxicity in primary cerebellar neurons in a polyglutamine length dependent manner in the context of full-length ataxin-7. Our results add ataxin-7 to a growing list of polyglutamine disease proteins that are capable of nuclear shuttling, and we define an activity of ataxin-7 in the STAGA complex of trafficking between the nucleus and cytoplasm. PMID- 16314425 TI - Acute in vivo effect of ethanol (binge drinking) on histone H3 modifications in rat tissues. AB - AIMS: To investigate the effect of acute in vivo administration of ethanol on acetylation or methylation of histone H3 at lysine9 in different tissues in rat. METHODS: Ethanol was injected into the stomach of Sprague-Dawley rats (8-weeks old) using blunt tipped needle. The rats were divided into three groups based on ethanol exposure times (1, 3, and 12 h). Each group was compared with water injected control group. The tissues from 14 different organs were removed. We essentially used similar type of protocol, tissue homogenization method, and sucrose density gradient centrifugation for isolation of nuclei with only minor modifications for some organs. Histone was isolated from the nuclei using acid extraction method. Acetylation of histone H3 at lysine9 (Ac-H3-lys9) and methylation of histone H3 at lysine9 (Me-H3-lys9) were analysed by western blotting. RESULTS: Effect of ethanol on Ac-H3-lys9 was investigated in 11 out of 14 rat tissues. In liver, we observed an increase in Ac-H3-lys9 with maximal increase of approximately 6-fold after 12 h exposure. Lung also showed approximately 3-fold increase. In spleen, ethanol-induced Ac-H3-lys9 in all three ethanol-treated groups with similar increase (1.5- to 1.6-fold). Testes showed significant increase (3-fold increase) of Ac-H3-lys9 only at 1 h ethanol exposure. Ethanol had no affect on Ac-H3-lys9 in other tissues: kidney, brain, heart, stomach, colorectum, pancreas, and vessels. Ethanol had little effect on Me-H3-lys9 in all rat tissues examined. CONCLUSIONS: After in vivo administration of ethanol, analogous to binge drinking condition, the acetylations of H3-lys9 in rat tissues are not universal but tissue-specific events with different patterns of responses. Ac-H3-Lys9 in liver, lung, and spleen were significantly affected and it was demonstrated that ethanol causes this epigenetic alteration in rat tissues selectively. PMID- 16314426 TI - NETER alcoholic typology (NAT). AB - AIMS: To establish an alcohol-dependent drinker's clinical typology, based on patients attending the Alcoholism Unit of Santa Maria's General Hospital in Lisbon, Portugal. METHODS: A multivariate statistical analysis was used to extract the typology solution. RESULTS: We obtained five factors: Anxiopathic, typifies anxious functioning; Heredopathic, congregates familiar and genetic influences on alcoholism; Thimopathic, typified by affective symptomatology; Sociopathic, characterized by disruptive behaviours under alcohol influence; and Adictopathic, isolates younger individuals who consume alcohol and other types of psychoactive substances. CONCLUSIONS: There are increasingly alcoholic polymorphic subtypes derived from the interactive complexity between genetic/family and psychosocial factors. PMID- 16314427 TI - Individuals with excessive alcohol intake recruited by advertisement: demographic and clinical characteristics. AB - AIMS: Studies have shown that most individuals with alcohol problems have never received any treatment for their alcoholism. The purpose of the present study was to describe demographic and clinical characteristics in male individuals with excessive alcohol intake who were recruited by advertisements. These characteristics were compared between individuals with or without prior treatment histories. METHODS: Subjects (n = 367) responded to the advertisements in a regional daily newspaper and called the investigators. A structured interview was performed and a complete dataset of demographic and clinical information was collected in 342 individuals. RESULTS: Individuals with no prior treatment history (n = 238) were found to be more often cohabitant, employed, and they reported fewer on-going psychiatric symptoms than individuals with treatment histories (n = 104). CONCLUSION: Since individuals with no prior treatment history seldom experience psychiatric symptoms, they are less likely to seek treatment in the health care system. It is therefore of importance to find ways to reach this 'hidden' group early with excessive alcohol consumption. One way to do so might be via alcohol treatment programs at working places since the majority of them are employed. PMID- 16314428 TI - RanBP3 enhances nuclear export of active (beta)-catenin independently of CRM1. AB - beta-Catenin is the nuclear effector of the Wnt signaling cascade. The mechanism by which nuclear activity of beta-catenin is regulated is not well defined. Therefore, we used the nuclear marker RanGTP to screen for novel nuclear beta catenin binding proteins. We identified a cofactor of chromosome region maintenance 1 (CRM1)-mediated nuclear export, Ran binding protein 3 (RanBP3), as a novel beta-catenin-interacting protein that binds directly to beta-catenin in a RanGTP-stimulated manner. RanBP3 inhibits beta-catenin-mediated transcriptional activation in both Wnt1- and beta-catenin-stimulated human cells. In Xenopus laevis embryos, RanBP3 interferes with beta-catenin-induced dorsoventral axis formation. Furthermore, RanBP3 depletion stimulates the Wnt pathway in both human cells and Drosophila melanogaster embryos. In human cells, this is accompanied by an increase of dephosphorylated beta-catenin in the nucleus. Conversely, overexpression of RanBP3 leads to a shift of active beta-catenin toward the cytoplasm. Modulation of beta-catenin activity and localization by RanBP3 is independent of adenomatous polyposis coli protein and CRM1. We conclude that RanBP3 is a direct export enhancer for beta-catenin, independent of its role as a CRM1-associated nuclear export cofactor. PMID- 16314429 TI - Self-organization of an acentrosomal microtubule network at the basal cortex of polarized epithelial cells. AB - Mechanisms underlying the organization of centrosome-derived microtubule arrays are well understood, but less is known about how acentrosomal microtubule networks are formed. The basal cortex of polarized epithelial cells contains a microtubule network of mixed polarity. We examined how this network is organized by imaging microtubule dynamics in acentrosomal basal cytoplasts derived from these cells. We show that the steady-state microtubule network appears to form by a combination of microtubule-microtubule and microtubule-cortex interactions, both of which increase microtubule stability. We used computational modeling to determine whether these microtubule parameters are sufficient to generate a steady-state acentrosomal microtubule network. Microtubules undergoing dynamic instability without any stabilization points continuously remodel their organization without reaching a steady-state network. However, the addition of increased microtubule stabilization at microtubule-microtubule and microtubule cortex interactions results in the rapid assembly of a steady-state microtubule network in silico that is remarkably similar to networks formed in situ. These results define minimal parameters for the self-organization of an acentrosomal microtubule network. PMID- 16314430 TI - Mutants in trs120 disrupt traffic from the early endosome to the late Golgi. AB - Transport protein particle (TRAPP), a large complex that mediates membrane traffic, is found in two forms (TRAPPI and -II). Both complexes share seven subunits, whereas three subunits (Trs130p, -120p, and -65p) are specific to TRAPPII. Previous studies have shown that mutations in the TRAPPII-specific gene trs130 block traffic through or from the Golgi. Surprisingly, we report that mutations in trs120 do not block general secretion. Instead, trs120 mutants accumulate aberrant membrane structures that resemble Berkeley bodies and disrupt the traffic of proteins that recycle through the early endosome. Mutants defective in recycling also display a defect in the localization of coat protein I (COPI) subunits, implying that Trs120p may participate in a COPI-dependent trafficking step on the early endosomal pathway. Furthermore, we demonstrate that Trs120p largely colocalizes with the late Golgi marker Sec7p. Our findings imply that Trs120p is required for vesicle traffic from the early endosome to the late Golgi. PMID- 16314431 TI - Quantitative elucidation of a distinct spatial gradient-sensing mechanism in fibroblasts. AB - Migration of eukaryotic cells toward a chemoattractant often relies on their ability to distinguish receptor-mediated signaling at different subcellular locations, a phenomenon known as spatial sensing. A prominent example that is seen during wound healing is fibroblast migration in platelet-derived growth factor (PDGF) gradients. As in the well-characterized chemotactic cells Dictyostelium discoideum and neutrophils, signaling to the cytoskeleton via the phosphoinositide 3-kinase pathway in fibroblasts is spatially polarized by a PDGF gradient; however, the sensitivity of this process and how it is regulated are unknown. Through a quantitative analysis of mathematical models and live cell total internal reflection fluorescence microscopy experiments, we demonstrate that PDGF detection is governed by mechanisms that are fundamentally different from those in D. discoideum and neutrophils. Robust PDGF sensing requires steeper gradients and a much narrower range of absolute chemoattractant concentration, which is consistent with a simpler system lacking the feedback loops that yield signal amplification and adaptation in amoeboid cells. PMID- 16314432 TI - Terminating Wnt signals: a novel nuclear export mechanism targets activated (beta)-catenin. AB - Nuclear targeting of beta-catenin is an obligatory step in Wnt signal transduction, but the factors that control import and export remain to be clarified. In this issue, Hendriksen et al. (p. 785) show that the RanBP3 export factor antagonizes beta-catenin/T cell factor (TCF) transcription by targeting the signaling-competent form of beta-catenin. We speculate that cells may use multiple export mechanisms to inhibit beta-catenin signaling in different ways. PMID- 16314433 TI - Adenovirus serotype 5 E1A sensitizes tumor cells to NKG2D-dependent NK cell lysis and tumor rejection. AB - The expression of the Adenovirus serotype 5 (Ad5) E1A oncogene sensitizes tumor cells to natural killer (NK) cell-mediated killing and tumor rejection in vivo. These effects are dependent on the ability of E1A to bind the transcriptional coadaptor protein p300. To test the hypothesis that E1A up-regulates ligands recognized by the NKG2D-activating receptor, we stably transfected the highly tumorigenic mouse fibrosarcoma cell line MCA-205 with Ad5-E1A or a mutant form of E1A that does not interact with p300 (E1A-Deltap300). Ad5-E1A, but not E1A Deltap300, up-regulated the expression of the NKG2D ligand retinoic acid early inducible (RAE)-1, but not murine ULBP-like transcript 1, another NKG2D ligand, in four independently derived MCA-205 transfectants. The up-regulation of RAE-1 by E1A targeted MCA-205 tumor cells to lysis by NK cells, resulting in NKG2D dependent tumor rejection in vivo. Moreover, the up-regulation of NKG2D ligands by E1A was not limited to mouse tumor cells, as E1A also increased the expression of NKG2D ligands on primary baby mouse kidney cells, human MB435S breast cancer cells, and human H4 fibrosarcoma cells. PMID- 16314434 TI - Immunostimulatory oligonucleotides block allergic airway inflammation by inhibiting Th2 cell activation and IgE-mediated cytokine induction. AB - A single treatment with a CpG-containing immunostimulatory DNA sequence (ISS) given before allergen challenge can inhibit T helper type 2 cell (Th2)-mediated airway responses in animal models of allergic asthma; however, the mechanism of this inhibition remains largely undefined. Here, we demonstrate that airway delivery of ISS before allergen challenge in Th2-primed mice acts in two distinct ways to prevent the allergic responses to this challenge. The first is to prevent induction of cytokines from allergen-specific Th2 cells, as demonstrated by the nearly complete inhibition of Th2 cytokine production, Th2-dependent functional responses, and gene induction patterns. ISS inhibits the Th2 response by rendering lung antigen-presenting cells (APCs) unable to effectively present antigen to Th2 cells, but not to Th1 cells. This loss of APC function correlates with a reduced expression of costimulatory molecules, including programmed cell death ligand (PD-L)1, PD-L2, CD40, CD80, CD86, and inducible T cell costimulator, and of major histocompatibility complex class II on CD11c(+ )APCs from the airways of ISS-treated mice. The second important action of ISS is inhibition of immunoglobulin E-dependent release of Th2 cytokines, especially interleukin 4, from basophils and/or mast cells in the airways of Th2-primed mice. Thus, inhibition by ISS of allergic responses can be explained by two novel mechanisms that culminate in the inhibition of the principal sources of type 2 cytokines in the airways. PMID- 16314435 TI - Resolution of airway inflammation and hyperreactivity after in vivo transfer of CD4+CD25+ regulatory T cells is interleukin 10 dependent. AB - Deficient suppression of T cell responses to allergen by CD4+CD25+ regulatory T cells has been observed in patients with allergic disease. Our current experiments used a mouse model of airway inflammation to examine the suppressive activity of allergen-specific CD4+CD25+ T cells in vivo. Transfer of ovalbumin (OVA) peptide-specific CD4+CD25+ T cells to OVA-sensitized mice reduced airway hyperreactivity (AHR), recruitment of eosinophils, and T helper type 2 (Th2) cytokine expression in the lung after allergen challenge. This suppression was dependent on interleukin (IL) 10 because increased lung expression of IL-10 was detected after transfer of CD4+CD25+ T cells, and regulation was reversed by anti IL-10R antibody. However, suppression of AHR, airway inflammation, and increased expression of IL-10 were still observed when CD4+CD25+ T cells from IL-10 gene deficient mice were transferred. Intracellular cytokine staining confirmed that transfer of CD4+CD25+ T cells induced IL-10 expression in recipient CD4+ T cells, but no increase in IL-10 expression was detected in airway macrophages, dendritic cells, or B cells. These data suggest that CD4+CD25+ T cells can suppress the Th2 cell-driven response to allergen in vivo by an IL-10-dependent mechanism but that IL-10 production by the regulatory T cells themselves is not required for such suppression. PMID- 16314436 TI - Endomucin, a CD34-like sialomucin, marks hematopoietic stem cells throughout development. AB - To detect as yet unidentified cell-surface molecules specific to hematopoietic stem cells (HSCs), a modified signal sequence trap was successfully applied to mouse bone marrow (BM) CD34(-)c-Kit(+)Sca-1(+)Lin(-) (CD34(-)KSL) HSCs. One of the identified molecules, Endomucin, is an endothelial sialomucin closely related to CD34. High-level expression of Endomucin was confined to the BM KSL HSCs and progenitor cells, and, importantly, long-term repopulating (LTR)-HSCs were exclusively present in the Endomucin(+)CD34(-)KSL population. Notably, in the yolk sac, Endomucin expression separated multipotential hematopoietic cells from committed erythroid progenitors in the cell fraction positive for CD41, an early embryonic hematopoietic marker. Furthermore, developing HSCs in the intraembryonic aorta-gonad-mesonephros (AGM) region were highly enriched in the CD45(-)CD41(+)Endomucin(+) fraction at day 10.5 of gestation (E10.5) and in the CD45(+)CD41(+)Endomucin(+) fraction at E11.5. Detailed analyses of these fractions uncovered drastic changes in their BM repopulating capacities as well as in vitro cytokine responsiveness within this narrow time frame. Our findings establish Endomucin as a novel cell-surface marker for LTR-HSCs throughout development and provide a powerful tool in understanding HSC ontogeny. PMID- 16314437 TI - CD4+CD25+ T cells protect against experimentally induced asthma and alter pulmonary dendritic cell phenotype and function. AB - The role of natural CD4+CD25+ regulatory T (T reg) cells in the control of allergic asthma remains poorly understood. We explore the impact of T reg cell depletion on the allergic response in mice susceptible (A/J) or comparatively resistant (C3H) to the development of allergen-induced airway hyperresponsiveness (AHR). In C3H mice, anti-CD25-mediated T reg cell depletion before house dust mite treatment increased several features of the allergic diathesis (AHR, eosinophilia, and IgE), which was concomitant with elevated T helper type 2 (Th2) cytokine production. In similarly T reg cell-depleted A/J mice, we observed a moderate increase in airway eosinophilia but no effects on AHR, IgE levels, or Th2 cytokine synthesis. As our experiments suggested that T reg cell depletion in C3H mice before sensitization was sufficient to enhance the allergic phenotype, we characterized dendritic cells (DCs) in T reg cell-depleted C3H mice. T reg cell-depleted mice had increased numbers of pulmonary myeloid DCs with elevated expression of major histocompatibility complex class II, CD80, and CD86. Moreover, DCs from T reg cell-depleted mice demonstrated an increased capacity to stimulate T cell proliferation and Th2 cytokine production, which was concomitant with reduced IL-12 expression. These data suggest that resistance to allergen driven AHR is mediated in part by CD4+CD25+ T reg cell suppression of DC activation and that the absence of this regulatory pathway contributes to susceptibility. PMID- 16314438 TI - Blimp-1 is required for maintenance of long-lived plasma cells in the bone marrow. AB - Long-lived plasma cells, residing primarily in the bone marrow, continuously secrete antibody and provide an important component of humoral memory. However, when such cells secrete autoantibodies or become transformed, they can be pathogenic. We have shown recently that the transcriptional repressor B lymphocyte-induced maturation protein 1 (Blimp-1) is required for the formation of plasma cells. To determine what role Blimp-1 might play in maintenance of plasma cells, we generated mice in which the gene encoding Blimp-1 could be deleted in an inducible manner. Deletion of Blimp-1 either in vitro or in vivo leads to loss of previously formed B220(LO)CD138(HI) plasma cells. Using BrdU incorporation, we confirmed that Blimp-1 is required for the maintenance of nondividing, long-lived plasma cells in the bone marrow. Blimp-1 is also required for long-term maintenance of antigen-specific immunoglobulin in serum. Thus Blimp 1 is required not only for the formation but also for the maintenance of long lived plasma cells. This finding provides the possibility of new drug design strategies for autoimmunity and multiple myeloma focused on blocking Blimp-1 expression or activity. PMID- 16314439 TI - Structural basis for CD1d presentation of a sulfatide derived from myelin and its implications for autoimmunity. AB - Sulfatide derived from the myelin stimulates a distinct population of CD1d restricted natural killer T (NKT) cells. Cis-tetracosenoyl sulfatide is one of the immunodominant species in myelin as identified by proliferation, cytokine secretion, and CD1d tetramer staining. The crystal structure of mouse CD1d in complex with cis-tetracosenoyl sulfatide at 1.9 A resolution reveals that the longer cis-tetracosenoyl fatty acid chain fully occupies the A' pocket of the CD1d binding groove, whereas the sphingosine chain fills up the F' pocket. A precise hydrogen bond network in the center of the binding groove orients and positions the ceramide backbone for insertion of the lipid tails in their respective pockets. The 3'-sulfated galactose headgroup is highly exposed for presentation to the T cell receptor and projects up and away from the binding pocket due to its beta linkage, compared with the more intimate binding of the alpha-glactosyl ceramide headgroup to CD1d. These structure and binding data on sulfatide presentation by CD1d have important implications for the design of therapeutics that target T cells reactive for myelin glycolipids in autoimmune diseases of the central nervous system. PMID- 16314440 TI - TNF-alpha-induced chemokine production and apoptosis in human neural precursor cells. AB - Recent studies have shown that proinflammatory cytokines damage rodent neural precursor cells (NPCs), a source of self-renewing, multipotent cells that play an important role in the developing as well as adult brain. In this study, the effects of tumor necrosis factor alpha (TNF-alpha) on cytokine and chemokine production by human NPCs (>98% nestin- and >90% A2B5-positive), obtained from 6- to 8-week-old fetal brain specimens, were evaluated. NPCs stimulated with this proinflammatory cytokine were found to produce abundant amounts of the chemokines monocyte chemoattractant protein 1 (MCP-1)/CC chemokine ligand 2 (CCL2) and interferon-inducible protein 10 (IP-10)/CXC chemokine ligand 10 (CXCL10) in a time- and concentration-dependent manner. TNF-alpha treatment also induced NPC apoptosis. Receptors for TNF [TNFRI (p55) and TNFRII (p75)] mRNA were constitutively expressed on NPCs. However, only TNFRI was involved in TNF-alpha induced chemokine production and apoptosis by NPCs, as anti-TNFRI but not anti TNFRII antibodies blocked the stimulatory effect. TNF-alpha treatment induced p38 mitogen-activated protein kinase (MAPK) phosphorylation in NPCs, and SB202190, an inhibitor of p38 MAPK, blocked TNF-alpha-induced chemokine production. Thus, this study demonstrated that NPCs constitutively express receptors for TNF-alpha, which when activated, trigger via a p38 MAPK signaling pathway production of two chemokines, MCP-1/CCL2 and IP-10/CXCL10, which are involved in infectious and inflammatory diseases of the brain. PMID- 16314441 TI - Ultrasound-accelerated tissue fixation/processing achieves superior morphology and macromolecule integrity with storage stability. AB - We demonstrate that high-frequency and high-intensity ultrasound (US) can be applied to both tissue fixation and tissue processing to complete the conventional overnight formalin-fixation and paraffin-embedding (FFPE) procedures within 1 hr. US-facilitated FFPE retains superior tissue morphology and long-term room temperature storage stability than conventional FFPE. There is less alteration of protein antigenicity after US-FFPE preservation so that rapid immunohistochemical reactions occur with higher sensitivity and intensity, reducing the need for antigen retrieval pretreatment. US-FFPE tissues present storage stability so that room temperature storage up to 7 years does not significantly affect tissue morphology, protein antigenic properties, RNA distribution, localization, and quantitation. In addition, during fixation, tissue displays physical changes that can be monitored and reflected as changes in transmission US signals. As far as we know, this is the first effort to monitor tissue physical changes during fixation. Further study of this phenomenon may provide a method to control and to monitor the level of fixation for quality controls. The mechanism of less alteration of protein antigenicity by US-FFPE was discussed. PMID- 16314442 TI - NF1 gene expression in mouse fracture healing and in experimental rat pseudarthrosis. AB - Neurofibromatosis type 1 (NF1) is an inherited disease with an incidence of about 1:3000 worldwide. Approximately half of all patients with NF1 present osseous manifestations, which can vary from mild to severely debilitating changes such as congenital pseudarthrosis. In the present study, fracture healing of mouse tibia was followed and specimens were collected 5, 9, 14, and 22 days postoperatively. Experimental pseudarthrosis of rat was followed up to 15 weeks postoperatively. In situ hybridization and immunohistochemistry were used to demonstrate expression of NF1 tumor suppressor and phosphorylated p44/42 mitogen-activated protein kinase (MAPK), an indicator of the Ras-MAPK pathway. The results showed that ossified callus was formed in mouse fracture 22 days after the operation. The final outcome of rat pseudarthrosis was detected 9 weeks after the operation, presenting abundant cartilaginous callus at the pseudarthrosis. NF1 gene expression was noted in the maturing and in the hypertrophic cartilages during normal mouse fracture healing, and in rat pseudarthrosis. Phosphorylated p44/42 MAPK was detected in a subpopulation of the hypertrophic chondrocytes in both models. Furthermore, positive labeling for NF1 mRNA and protein was detected in endothelium in both the pseudarthrosis and in the fracture. In conclusion, NF1 gene expression and function are needed for normal fracture healing, possibly restraining excessive Ras-MAPK pathway activation. PMID- 16314443 TI - Distribution of CIAPIN1 in normal fetal and adult human tissues. AB - CIAPIN1, a newly identified antiapoptotic molecule that plays an essential role in mouse definitive hematopoiesis, is considered a downstream effector of the receptor tyrosine kinase-Ras signaling pathway. Our previous studies have indicated that CIAPIN1 is involved in the development of multidrug resistance (MDR) in gastric cancer cells. However, the mechanism of CIAPIN1-mediated antiapoptosis and MDR has not been fully elucidated. To reveal the possible physiological role of CIAPIN1, we examined the expression and distribution of CIAPIN1 in fetal and adult human tissues using immunohistochemistry. We found that CIAPIN1 was ubiquitously distributed in fetal and adult tissues, and was localized in both the cytoplasm and the nucleus. The expression patterns of CIAPIN1 were similar in fetal and adult tissues, and was correlated with the previously described expression pattern of p21ras. These observations suggest that CIAPIN1 expression appears to be involved in cell differentiation, and that it might exert universal and possibly important physiological functions under the regulation of Ras in humans. PMID- 16314444 TI - 25-hydroxyvitamin D3 1alpha-hydroxylase and vitamin D receptor expression in papillary thyroid carcinoma. AB - Vitamin D receptor (VDR) and 25-hydroxyvitamin D3 1-alpha-hydroxylase expression have recently been shown to be upregulated in several tumors and thought to represent an important endogenous response to tumor progression. Little is known about the expression of these proteins in thyroid carcinoma, although previous reports have documented evidence of the biological effect of vitamin D in thyroid cells. Using paraffin-embedded and frozen sections of papillary thyroid carcinoma, we utilized real-time quantitative RT-PCR and immunohistochemistry to characterize the expression of VDR and 1-alpha-hydroxylase in thyroid follicular cells, with special emphasis on papillary thyroid carcinoma (PTC). VDR and 1 alpha-hydroxylase expression were increased in PTC compared with normal thyroid tissue and especially high in areas of lymphocyte infiltration. Expression of VDR and 1-alpha-hydroxylase in PTC may be compatible with an overall favorable prognosis for this tumor type and may constitute important prerequisites for using vitamin D and/or vitamin D analogs in the treatment of PTC. PMID- 16314445 TI - Alterations of phosphorylation state of connexin 43 during hypoxia and reoxygenation are associated with cardiac function. AB - Gap junctions formed by connexins mediate cell-cell communication by electrical and chemical coupling. Recently, it has been shown that alterations in the phosphorylation state of the connexins result in functional alteration of cell cell communication through gap junctions. Therefore, we focused on the association of alterations of phosphorylation state of connexin 43 (Cx43) with cardiac function in vivo. Rat hearts were transferred to Langendorff apparatus and submitted to hypoxia and then reoxygenated. In the control heart, Cx43 was phosphorylated and located at the intercalated disk. When the hearts were subjected to hypoxia, Cx43 at gap junctions was dephosphorylated and changed its localization to the entire plasma membrane. The area of cardiomyocytes stained with anti-phosphorylated Cx43 antibody was decreased in a time-dependent manner. Immunoblot data supported the decrease of phosphorylated Cx43 during hypoxia. ZO 1 did not change its localization at the intercalated disk during the hypoxic period. We also found that the area occupied by dephosphorylated Cx43 was correlated with the decrease of percent of rate-pressure product. These data indicate that dephosphorylation and redistribution of Cx43 is an early sign of cardiac injury after hypoxia. Detection of dephosphorylated Cx43 may serve as a diagnostic tool for examining ischemic heart disease. PMID- 16314446 TI - Cell-specific subcellular localization of soluble epoxide hydrolase in human tissues. AB - Soluble epoxide hydrolase (sEH) is a phase-I xenobiotic metabolizing enzyme having both an N-terminal phosphatase activity and a C-terminal epoxide hydrolase activity. Endogenous hydrolase substrates include arachidonic acid epoxides, which have been involved in regulating blood pressure and inflammation. The subcellular localization of sEH has been controversial. Earlier studies using mouse and rat liver suggested that sEH may be cytosolic and/or peroxisomal. In this study we applied immunofluorescence and confocal microscopy using markers for different subcellular compartments to evaluate sEH colocalization in an array of human tissues. Results showed that sEH is both cytosolic and peroxisomal in human hepatocytes and renal proximal tubules and exclusively cytosolic in other sEH-containing tissues such as pancreatic islet cells, intestinal epithelium, anterior pituitary cells, adrenal gland, endometrium, lymphoid follicles, prostate ductal epithelium, alveolar wall, and blood vessels. sEH was not exclusively peroxisomal in any of the tissues evaluated. Our data suggest that human sEH subcellular localization is tissue dependent, and that sEH may have tissue- or cell-type-specific functionality. To our knowledge, this is the first report showing the subcellular localization of sEH in a wide array of human tissues. PMID- 16314447 TI - Polar localizing class V myosin chitin synthases are essential during early plant infection in the plant pathogenic fungus Ustilago maydis. AB - Fungal chitin synthases (CHSs) form fibers of the cell wall and are crucial for substrate invasion and pathogenicity. Filamentous fungi contain up to 10 CHSs, which might reflect redundant functions or the complex biology of these fungi. Here, we investigate the complete repertoire of eight CHSs in the dimorphic plant pathogen Ustilago maydis. We demonstrate that all CHSs are expressed in yeast cells and hyphae. Green fluorescent protein (GFP) fusions to all CHSs localize to septa, whereas Chs5-GFP, Chs6-GFP, Chs7-yellow fluorescent protein (YFP), and Myosin chitin synthase1 (Mcs1)-YFP were found at growth regions of yeast-like cells and hyphae, indicating that they participate in tip growth. However, only the class IV CHS genes chs7 and chs5 are crucial for shaping yeast cells and hyphae ex planta. Although most CHS mutants were attenuated in plant pathogenicity, Deltachs6, Deltachs7, and Deltamcs1 mutants were drastically reduced in virulence. Deltamcs1 showed no morphological defects in hyphae, but Mcs1 became essential during invasion of the plant epidermis. Deltamcs1 hyphae entered the plant but immediately lost growth polarity and formed large aggregates of spherical cells. Our data show that the polar class IV CHSs are essential for morphogenesis ex planta, whereas the class V myosin-CHS is essential during plant infection. PMID- 16314448 TI - British Thoracic Society Winter Meeting 2005, London, England, United Kingdom, 7 9 December. Abstracts. PMID- 16314451 TI - MicroRNA function: multiple mechanisms for a tiny RNA? AB - MicroRNAs are sequence-specific regulators of post-transcriptional gene expression in many eukaryotes. They are believed to control the expression of thousands of target mRNAs, with each mRNA believed to be targeted by multiple microRNAs. Recent studies have uncovered various mechanisms by which microRNAs down-regulate their target mRNAs and have linked a well-known subcellular structure, the cytoplasmic processing bodies (PBs) to the microRNA pathway. The finding that microRNAs are misexpressed in cancers has reinforced the idea that their regulatory roles are very important. PMID- 16314452 TI - A pseudoknot in the 3' non-core region of the glmS ribozyme enhances self cleavage activity. AB - The recently described glmS ribozyme is a self-cleaving RNA sequence found in the 5' noncoding region of the transcript of the gene for glucosamine-6-phosphate (GlcN6P) synthase in many Gram-positive bacteria. This ribozyme is associated with the GlcN6P riboswitch, and ribozyme activity in response to binding of the metabolite, GlcN6P, is proposed to effect levels of gene expression. The previously defined core sequence of the GlcN6P-dependent ribozyme contained fewer than 80 nt of contiguous sequence, but a sequence containing conserved secondary structural features and encompassing the core was twice as long. Structural elements outside of the ribozyme core could contribute to ribozyme activity or participate in gene regulation as part of the expression platform or both. Here, a 174-nt transcript containing the Bacillus anthracis glmS ribozyme was used to examine the contribution of part of the non-core sequence to in vitro cleavage activity. The loop portion of hairpin loop 3, located just 3' of the ribozyme core, can potentially pair with a sequence approximately 80 nt downstream to form a pseudoknot tertiary interaction. Disruptive and compensatory mutations in the two duplex regions of the pseudoknot had effects on in vitro cleavage rates that support a role for the pseudoknot in enhanced ribozyme activity. Cleavage activity became less sensitive to disruptive mutations in the pseudoknot as MgCl(2) concentrations were raised from 2.5 to 10 mM, suggesting that one role of the pseudoknot could be to help stabilize the core structure. PMID- 16314453 TI - Ge-1 is a central component of the mammalian cytoplasmic mRNA processing body. AB - The mRNA processing body (P-body) is a cellular structure that regulates gene expression by degrading cytoplasmic mRNA. The objective of this study was to identify and characterize novel components of the mammalian P-body. Approximately 5% of patients with the autoimmune disease primary biliary cirrhosis have antibodies directed against this structure. Serum from one of these patients was used to identify a cDNA encoding Ge-1, a 1,401-amino-acid protein. Ge-1 contains an N-terminal WD 40 motif and C-terminal domains characterized by a repeating psi(X(2-3)) motif. Ge-1 co-localized with previously identified P-body components, including proteins involved in mRNA decapping (DCP1a and DCP2) and the autoantigen GW 182. The Ge-1 C-terminal domain was necessary and sufficient to target the protein to P-bodies. Following exposure of cells to oxidative stress, Ge-1-containing P-bodies were found adjacent to TIA-containing stress granules. During the recovery period, TIA returned to the nucleus while Ge-1 containing P-bodies localized to the perinuclear region. siRNA-mediated knock down of Ge-1 resulted in loss of P-bodies containing Ge-1, DCP1a, and DCP2. In contrast, Ge-1-containing P-bodies persisted despite knock-down of DCP2. Taken together, the results of this study show that Ge-1 is a central component of P bodies and suggest that Ge-1 may act prior to the 5(')-decapping step in mRNA degradation. PMID- 16314454 TI - The polypyrimidine tract binding protein is a monomer. AB - The polypyrimidine tract binding (PTB) protein is a potent regulator of alternative mRNA splicing. It also participates in other essential cellular functions, including translation initiation and polyadenylation. Several published reports have suggested that the protein forms a dimer in solution, a feature that has been widely incorporated into mechanistic models of protein function. However, recent studies have provided indications that full-length PTB is a monomer. Here we present new biophysical and biochemical evidence supporting the monomeric status of the protein. By use of blue-native polyacrylamide gel electrophoresis and size-exclusion chromatography, PTB was observed as a single molecular species under native reducing environments, though in oxidizing conditions, a larger protein species was also detected. Further analyses of wild type and mutant PTB molecules with SDS-PAGE and time-of-flight electrospray ionization mass spectroscopy confirmed these observations. They also identified the single reduced species as monomeric PTB and the higher-molecular-weight nonreduced species as disulphide-linked PTB dimer mediated by Cys23. Our results indicate that the use of oxidizing environments in previous studies is likely to have contributed to the mis-assignment of PTB as a dimer. Although purified PTB may form disulphide-linked dimers under these conditions, in the reducing intracellular environment the protein will be monomeric. These findings have implications for the construction of models of PTB function in regulating mRNA metabolism. PMID- 16314455 TI - Evidence for an RNA chaperone function of polypyrimidine tract-binding protein in picornavirus translation. AB - The cellular polypyrimidine tract-binding protein (PTB) is recruited by the genomic RNAs of picornaviruses to stimulate translation initiation at their internal ribosome entry site (IRES) elements. We investigated the contribution of the individual RNA recognition motif (RRM) domains of PTB to its interaction with the IRES of foot-and-mouth disease virus (FMDV). Using a native gel system, we found that PTB is a monomer, confirming recent reports that challenged the previous view that PTB is a dimer. Mapping the spatial orientation of PTB relative to the bound IRES RNA, we found that the two C-terminal RRM domains III and IV of PTB bind in an oriented way to the IRES. Domain III contacts the IRES stem-loop 2, while domain IV contacts the separate IRES 3' region. PTB domain I appears not to be involved directly in RNA binding, but domain II stabilizes the RNA binding conferred by domains III and IV. A PTB protein containing only these two C-terminal PTB domains is sufficient to enhance the entry of initiation factor eIF4G to the IRES and stimulate IRES activity, and the long-lived PTB-IRES interaction stabilized by domain II is not a prerequisite for this function. Thus, PTB most likely acts as an RNA chaperone to stabilize IRES structure and, in that way, augment IRES activity. PMID- 16314456 TI - A consensus CaMK IV-responsive RNA sequence mediates regulation of alternative exons in neurons. AB - Neurons make extensive use of alternative pre-mRNA splicing to regulate gene expression and diversify physiological responses. We showed previously in a pituitary cell line that the Ca(++)/calmodulin-dependent protein kinase CaMK IV specifically repressed splicing of the BK channel STREX exon. This repression is dependent on a CaMK IV-responsive RNA element (CaRRE) within the STREX 3' splice site. Here, we report that similar Ca(++) regulation of splicing, mediated by L type calcium channels and CaM kinase IV, occurs in cultured neurons and in the brain. We identify a critical CaRRE motif (CACATNRTTAT) that is essential for conferring CaMK IV repression on an otherwise constitutive exon. Additional Ca(++)-regulated exons that carry this consensus sequence are also identified in the human genome. Thus, the Ca(++)/CaMK IV pathway in neurons controls the alternative splicing of a group of exons through this short CaRRE consensus sequence. The functions of some of these exons imply that splicing control through the CaMK IV pathway will alter neuronal activity. PMID- 16314457 TI - Arginine-rich motifs present multiple interfaces for specific binding by RNA. AB - A number of proteins containing arginine-rich motifs (ARMs) are known to bind RNA and are involved in regulating RNA processing in viruses and cells. Using automated selection methods we have generated a number of aptamers against ARM peptides from various natural proteins. Aptamers bind tightly to their cognate ARMs, with K(d) values in the nanomolar range, and frequently show no propensity to bind to other ARMs or even to single amino acid variants of the cognate ARM. However, at least some anti-ARM aptamers can cross-recognize a limited set of other ARMs, just as natural RNA-binding sites have been shown to exhibit so called "chameleonism." We expand upon the number of examples of cross-recognition and, using mutational and circular dichroism (CD) analyses, demonstrate that there are multiple mechanisms by which RNA ligands can cross-recognize ARMs. These studies support a model in which individual arginine residues govern binding to an RNA ligand, and the inherent flexibility of the peptide backbone may make it possible for "semi-specific" recognition of a discrete set of RNAs by a discrete set of ARM peptides and proteins. PMID- 16314458 TI - Biochemical analysis of the EJC reveals two new factors and a stable tetrameric protein core. AB - The multiprotein exon junction complex (EJC) is deposited on mRNAs upstream of exon-exon junctions as a consequence of pre-mRNA splicing. In mammalian cells, this complex serves as a key modulator of spliced mRNA metabolism. To date, neither the complete composition nor the exact assembly pathway of the EJC has been entirely elucidated. Using in vitro splicing and a two-step chromatography procedure, we have purified the EJC and analyzed its components by mass spectrometry. In addition to finding most of the known EJC factors, we identified two novel EJC components, Acinus and SAP18. Heterokaryon analysis revealed that SAP18 is a shuttling protein whereas Acinus is restricted to the nucleus. In MS2 tethering assays Acinus stimulated gene expression at the RNA level, while MLN51, another EJC factor, stimulated mRNA translational efficiency. Using tandem affinity purification (TAP) of proteins overexpressed in HeLa cells, we demonstrated that Acinus binds directly to another EJC component, RNPS1, while stable association of SAP18 to form the trimeric apoptosis and splicing associated protein (ASAP) complex requires both Acinus and RNPS1. Using the same methodology, we further identified what appears to be the minimal stable EJC core, a heterotetrameric complex consisting of eIF4AIII, Magoh, Y14, and MLN51. PMID- 16314459 TI - Accessibility of 18S rRNA in human 40S subunits and 80S ribosomes at physiological magnesium ion concentrations--implications for the study of ribosome dynamics. AB - Protein biosynthesis requires numerous conformational rearrangements within the ribosome. The structural core of the ribosome is composed of RNA and is therefore dependent on counterions such as magnesium ions for function. Many steps of translation can be compromised or inhibited if the concentration of Mg(2+) is too low or too high. Conditions previously used to probe the conformation of the mammalian ribosome in vitro used high Mg(2+) concentrations that we find completely inhibit translation in vitro. We have therefore probed the conformation of the small ribosomal subunit in low concentrations of Mg(2+) that support translation in vitro and compared it with the conformation of the 40S subunit at high Mg(2+) concentrations. In low Mg(2+) concentrations, we find significantly more changes in chemical probe accessibility in the 40S subunit due to subunit association or binding of the hepatitis C internal ribosomal entry site (HCV IRES) than had been observed before. These results suggest that the ribosome is more dynamic in its functional state than previously appreciated. PMID- 16314460 TI - Complete MALDI-ToF MS analysis of cross-linked peptide-RNA oligonucleotides derived from nonlabeled UV-irradiated ribonucleoprotein particles. AB - Protein-RNA cross-linking combined with mass spectrometry is a powerful tool to elucidate hitherto non-characterized protein-RNA contacts in ribonucleoprotein particles, as, for example, within spliceosomes. Here, we describe an improved methodology for the sequence analysis of purified peptide-RNA oligonucleotide cross-links that is based solely on MALDI-ToF mass spectrometry. The utility of this methodology is demonstrated on cross-links isolated from UV-irradiated spliceosomal particles; these were (1) [15.5K-61 K-U4 atac] small nuclear ribonucleoprotein (snRNP) particles prepared by reconstitution in vitro, and (2) U1 snRNP particles purified from HeLa cells. We show that the use of 2',4',6' trihydroxyacetophenone (THAP) as MALDI matrix allows analysis of cross-linked peptide-RNA oligonucleotides in the reflectron mode at high resolution, enabling sufficient accuracy to assign unambiguously cross-linked RNA sequences. Most important, post-source decay (PSD) analysis under these conditions was successfully applied to obtain sequence information about the cross-linked peptide and RNA moieties within a single spectrum, including the identification of the actual cross-linking site. Thus, in U4 atac snRNA we identified His 270 in the spliceosomal U4/U6 snRNP-specific protein 61 K (hPrp31p) cross-linked to U 44; in the U1 snRNP we show that Leu175 of the U1 snRNP-specific 70K protein is cross-linked to U 30 of U1 snRNA. This type of analysis is applicable to any type of RNP complex and may be expected to pave the way for the further analysis of protein-RNA complexes in much lower abundance and/or of cross-links that are obtained in low yield. PMID- 16314461 TI - Toward a genome-wide systems biology analysis of host-pathogen interactions in group A Streptococcus. AB - Genome-wide analysis of microbial pathogens and molecular pathogenesis processes has become an area of considerable activity in the last 5 years. These studies have been made possible by several advances, including completion of the human genome sequence, publication of genome sequences for many human pathogens, development of microarray technology and high-throughput proteomics, and maturation of bioinformatics. Despite these advances, relatively little effort has been expended in the bacterial pathogenesis arena to develop and use integrated research platforms in a systems biology approach to enhance our understanding of disease processes. This review discusses progress made in exploiting an integrated genome-wide research platform to gain new knowledge about how the human bacterial pathogen group A Streptococcus causes disease. Results of these studies have provided many new avenues for basic pathogenesis research and translational research focused on development of an efficacious human vaccine and novel therapeutics. One goal in summarizing this line of study is to bring exciting new findings to the attention of the investigative pathology community. In addition, we hope the review will stimulate investigators to consider using analogous approaches for analysis of the molecular pathogenesis of other microbes. PMID- 16314462 TI - Cathepsin deficiency as a model for neuronal ceroid lipofuscinoses. PMID- 16314463 TI - Cisplatin nephrotoxicity is critically mediated via the human organic cation transporter 2. AB - Cis-platin is an effective anti-neoplastic agent, but it is also highly nephrotoxic. Here, we clearly identify the human organic cation transporter 2 (hOCT2) as the critical transporter for cis-platin nephrotoxicity in isolated human proximal tubules and offer a potential mechanism for reducing nephrotoxicity in clinical practice. Interaction of cis-platin with hOCT2 in kidney or hOCT1 in liver was investigated with the fluorescent cation 4-[4 (dimethyl-amino)styril]-methylpyridinium in stably transfected HEK293 cells and for the first time in tissues physiologically expressing these transporters, human proximal tubules, and human hepatocyte couplets. Cis-platin (100 micromol/L) inhibited transport via hOCT2-HEK293 but not hOCT1-HEK293. In human proximal tubules cis-platin competed with basolateral organic cation transport, whereas it had no effect in tubules from a diabetic kidney or in hepatocytes. In hOCT2-HEK293 cells treated for 15 hours, incubation with cis-platin induced apoptosis, which was completely suppressed by contemporaneous incubation with the hOCT2 substrate cimetidine (100 micromol/L). These findings demonstrate that uptake of cis-platin is mediated by hOCT2 in renal proximal tubules, explaining its organ-specific toxicity. A combination of cis-platin with other substrates that compete for hOCT2 offers an effective option to decrease nephrotoxicity in the clinical setting. PMID- 16314464 TI - Role of Eotaxin-1 (CCL11) and CC chemokine receptor 3 (CCR3) in bleomycin-induced lung injury and fibrosis. AB - Eotaxin-1/CCL11 and its receptor CCR3 are involved in recruitment of eosinophils to diverse tissues, but their role in eosinophil recruitment in pulmonary fibrosis is unclear. The present study examined the pulmonary expression of CCL11 and CCR3 during bleomycin (blm)-induced lung injury and determined their importance in the recruitment of inflammatory cells and the development of lung fibrosis. In mice, blm induced a marked pulmonary expression of CCL11 and CCR3. Immunostaining for CCR3 revealed that this receptor was not only expressed by eosinophils but also by neutrophils. CCL11-deficient (CCL11(-/-)) mice developed significantly reduced pulmonary fibrosis. Expression of profibrotic cytokines such as transforming growth factor-beta1 was diminished in the absence of CCL11. Furthermore, increased lung expression of CCL11 significantly enhanced blm induced lung fibrosis and production of profibrotic cytokines. These effects were also associated with an increase of eosinophil and neutrophil pulmonary infiltration. In contrast, mice treated with neutralizing CCR3 antibodies developed significantly reduced pulmonary fibrosis, eosinophilia, neutrophilia, and expression of profibrotic cytokines. Together, these data suggest that CCL11 and CCR3 are important in the pulmonary recruitment of granulocytes and play significant pathogenic roles in blm-induced lung fibrosis. PMID- 16314465 TI - Cytoplasmic YY1 is associated with increased smooth muscle-specific gene expression: implications for neonatal pulmonary hypertension. AB - Immediately after birth the adluminal vascular SMCs of the pulmonary elastic arteries undergo transient actin cytoskeletal remodeling as well as cellular de differentiation and proliferation. Vascular smooth muscle phenotype is regulated by serum response factor, which is itself regulated in part by the negative regulator YY1. We therefore studied the subcellular localization of YY1 in arteries of normal newborn piglets and piglets affected by neonatal pulmonary hypertension. We found that YY1 localization changed during development and that expression of gamma-smooth muscle actin correlated with expression of cytoplasmic rather than nuclear YY1. Analysis of the regulation of YY1 localization in vitro demonstrated that polymerized gamma-actin sequestered EGFP-YY1 in the cytoplasm and that YY1 activation of c-myc promoter activity was inhibited by LIM kinase, which increases actin polymerization. Consistent with these data siRNA-mediated down-regulation of YY1 in C2C12 cells increased SM22-alpha expression and inhibited cell proliferation. Thus, actin polymerization controls subcellular YY1 localization, which contributes to vascular SMC proliferation and differentiation in normal pulmonary artery development. In the absence of actin depolymerization, YY1 does not relocate to the nucleus, and this lack of relocation may contribute to the pathobiology of pulmonary hypertension. PMID- 16314466 TI - Arterial macrophages and regenerating endothelial cells express P-selectin in atherosclerosis-prone apolipoprotein E-deficient mice. AB - P-selectin expression has been reported in platelets, endothelial cells, and vascular smooth muscle cells in response to vascular injury. Here, we report P selectin expression on macrophages in the arterial wall after carotid denudation injury and spontaneous atherosclerosis in atherosclerosis-prone apoE-deficient (apoE(-/-)) mice. Double-immunofluorescence staining revealed robust P-selectin expression in macrophage-rich regions of both denudation-induced carotid neointimal lesions and innominate atherosclerotic plaques. Co-localization of P selectin with macrophages was verified at the single cell level using double immunostaining plus 4,6-diamidino-2-phenylindole (for nuclei) counterstaining. No platelet staining was seen in association with the macrophage staining, excluding platelet contamination. Furthermore, P-selectin mRNA expression was readily detectable in macrophage-rich plaques of atherosclerotic innominate arteries and blood monocyte-derived macrophages from apoE(-/-) mice. Strong P-selectin expression was also seen in the areas of regenerated endothelium after arterial injury. In addition, co-localization of P-selectin with vascular smooth muscle cells was readily observed in denudation-injured carotid arteries at 7 and 14 days. We conclude that macrophages in carotid injury-induced neointimal lesions and spontaneous atherosclerotic plaques of the innominate artery acquire the ability to express P-selectin, as does regenerating endothelium. These findings provide a potential new paradigm in macrophage-mediated vascular inflammation, atherosclerosis, and neointimal hyperplasia after arterial injury. PMID- 16314467 TI - Tissue regeneration using macrophage migration inhibitory factor-impregnated gelatin microbeads in cutaneous wounds. AB - Migration inhibitory factor (MIF) responds to tissue damage and regulates inflammatory and immunological processes. To elucidate the function of MIF in cutaneous wound healing, we analyzed MIF knockout (KO) mice. After the excision of wounds from the dorsal skin of MIF KO and wild-type (WT) mice, healing was significantly delayed in MIF KO mice compared to WT mice. Lipopolysaccharide treatment significantly increased [(3)H]thymidine uptake in WT mouse fibroblasts compared to MIF KO mouse fibroblasts. Furthermore, there was a significant reduction in fibroblast and keratinocyte migration observed in MIF KO mice after 1-oleoyl-2-lysophosphatidic acid treatment. We subsequently examined whether MIF impregnated gelatin slow-release microbeads could accelerate skin wound healing. Injection of more than 1.5 microg/500 microl of MIF-impregnated gelatin microbeads around a wound edge accelerated wound healing compared to a single MIF injection without the use of microbeads. MIF-impregnated gelatin microbeads also accelerated skin wound healing in C57BL/6 mice and diabetic db/db mice. Furthermore, incorporating MIF-impregnated gelatin microbeads into an artificial dermis implanted into MIF KO mice accelerated procollagen production and capillary formation. These findings suggest that MIF is crucial in accelerating cutaneous wound healing and that MIF-impregnated gelatin microbeads represent a promising treatment to facilitate skin wound healing. PMID- 16314468 TI - Novel mechanisms of target cell death and survival and of therapeutic action of IVIg in Pemphigus. AB - Pemphigus vulgaris (PV) is a potentially lethal mucocutaneous blistering disease characterized by cell-cell detachment within the stratified epithelium (acantholysis) caused by IgG autoantibodies. Intravenous immunoglobulin (IVIg) therapy effectively treats PV, but the mechanism is not fully understood. To further understand acantholysis and the efficacy of IVIg, we measured effects of IgG fractions from PV patients on keratinocyte death processes. Using IgGs from representative PV patients who improved with IVIg, we identified apoptotic and oncotic signaling pathways in in vitro and in vivo PV models. We identified two groups of PV patients, each producing autoantibodies activating predominantly either apoptotic or oncotic cell death pathway. Experimental treatments with caspase 3 or calpain inhibitors demonstrated that PV IgGs induced acantholysis through both pathways. Upstream, the apoptotic signaling involved activation of caspases 8 and 3 and up-regulation of Fas ligand mRNA, whereas calpain-mediated cell death depended on elevated intracellular free Ca(2+). IVIg reduced PV IgG mediated acantholysis and cell death and up-regulated the caspase inhibitor FLIP and the calpain inhibitor calpastatin. These results indicate that in different PV patients, IgG-induced acantholysis proceeds predominantly via distinct, yet complementary, pathways of programmed cell death differentially mediated by apoptosis and oncosis effectors, with IVIg protecting target cells by up regulating endogenous caspase and calpain inhibitors. PMID- 16314469 TI - Silencing of Fas, but not caspase-8, in lung epithelial cells ameliorates pulmonary apoptosis, inflammation, and neutrophil influx after hemorrhagic shock and sepsis. AB - Apoptosis and inflammation play an important role in the pathogenesis of direct/pulmonary acute lung injury (ALI). However, the role of the Fas receptor driven apoptotic pathway in indirect/nonpulmonary ALI is virtually unstudied. We hypothesized that if Fas or caspase-8 plays a role in the induction of indirect ALI, their local silencing using small interfering RNA (siRNA) should be protective in hemorrhage-induced septic ALI. Initially, as a proof of principle, green fluorescent protein-siRNA was administered intratracheally into transgenic mice overexpressing green fluorescent protein. Twenty-four hours after siRNA delivery, lung sections revealed a significant decrease in green fluorescence. Intratracheally administered Cy-5-labeled Fas-siRNA localized primarily in pulmonary epithelial cells. Intratracheal instillation of siRNA did not induce lung inflammation via toll-like receptor or protein kinase PKR pathways as assessed by lung tissue interferon-alpha, tumor necrosis factor-alpha, and interleukin (IL)-6 levels. Mice subjected to hemorrhagic shock and sepsis received either Fas-, caspase-8-, or control-siRNA intratracheally 4 hours after hemorrhage. Fas- or caspase-8-siRNA significantly reduced lung tissue Fas or caspase-8 mRNA, respectively. Only Fas-siRNA markedly diminished lung tissue tumor necrosis factor-alpha, IL-6, IL-10, interferon-gamma, IL-12, and caspase-3 activity. Fas-siRNA also preserved alveolar architecture and reduced lung neutrophil infiltration and pulmonary epithelial apoptosis. These data indicate the pathophysiological significance of Fas activation in nonpulmonary/shock induced ALI and the feasibility of intrapulmonary administration of anti apoptotic siRNA in vivo. PMID- 16314470 TI - Macrophage migration inhibitory factor: a central regulator of wound healing. AB - Age-associated differences in estrogen levels critically modify the cutaneous wound healing response. Using a microarray-based approach, we profiled changes in gene expression within the wounds of mice that were wild type or null for the pro inflammatory cytokine macrophage migration inhibitory factor (MIF) in the presence or absence of estrogen. This experimental design identified more than 600 differentially expressed genes and established MIF as a key player in the wound healing process, regulating many novel repair/inflammation-associated gene targets. Moreover, MIF affected virtually all of the effects of reduced estrogen on wound repair. In humans, serum and wound levels of MIF increased with age and were strongly down-regulated by estrogen in vivo. Estrogen-regulated MIF transcription in vitro via a nuclear factor kappaB-dependent mechanism. These findings have wide-ranging implications for the many pathophysiological states in which MIF plays an important regulatory role and suggest a potential therapeutic role for MIF in modulating clinical conditions associated with age-related decline in estrogen levels. PMID- 16314471 TI - Biomimetic delivery of keratinocyte growth factor upon cellular demand for accelerated wound healing in vitro and in vivo. AB - Exogenous keratinocyte growth factor (KGF) significantly enhances wound healing, but its use is hampered by a short biological half-life and lack of tissue selectivity. We used a biomimetic approach to achieve cell-controlled delivery of KGF by covalently attaching a fluorescent matrix-binding peptide that contained two domains: one recognized by factor XIII and the other by plasmin. Modified KGF was incorporated into the fibrin matrix at high concentration in a factor XIII dependent manner. Cell-mediated activation of plasminogen to plasmin degraded the fibrin matrix and cleaved the peptides, releasing active KGF to the local microenvironment and enhancing epithelial cell proliferation and migration. To demonstrate in vivo effectiveness, we used a hybrid model of wound healing that involved transplanting human bioengineered skin onto athymic mice. At 6 weeks after grafting, the transplanted tissues underwent full thickness wounding and treatment with fibrin gels containing bound KGF. In contrast to topical KGF, fibrin-bound KGF persisted in the wounds for several days and was released gradually, resulting in significantly enhanced wound closure. A fibrinolytic inhibitor prevented this healing, indicating the requirement for cell-mediated fibrin degradation to release KGF. In conclusion, this biomimetic approach of localized, cell-controlled delivery of growth factors may accelerate healing of large full-thickness wounds and chronic wounds that are notoriously difficult to heal. PMID- 16314472 TI - Transforming growth factor-beta regulation of epithelial tight junction proteins enhances barrier function and blocks enterohemorrhagic Escherichia coli O157:H7 induced increased permeability. AB - Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is an enteric pathogen that causes potentially fatal symptoms after intimate adhesion, modulation of intestinal epithelial signal transduction, and alteration of epithelial function (eg, barrier disruption). Although the epithelial barrier is critical to gut homeostasis, only a few agents, such as transforming growth factor (TGF)-beta, can enhance or protect epithelial barrier function. Our aims were to delineate the mechanism(s) behind TGF-beta-induced barrier enhancement and to determine whether TGF-beta could prevent EHEC-induced barrier disruption. Using monolayers of the human T84 colonic epithelial cell line, we found that TGF-beta induced a significant increase in transepithelial electrical resistance (a measure of paracellular permeability) through activation of ERK MAPK and SMAD signaling pathways and up-regulation of the tight junction protein claudin-1. Additionally, TGF-beta pretreatment of epithelia blocked the decrease in transepithelial electrical resistance and the increase in transepithelial passage of [(3)H] mannitol caused by EHEC infection. EHEC infection was associated with reduced expression of zonula occludens-1, occludin, and claudin-2 (but not claudin-1 or claudin-4); TGF-beta pretreatment prevented these changes. These studies provide insight into EHEC pathogenesis by illustrating the mechanisms underlying TGF-beta induced epithelial barrier enhancement and identifying TGF-beta as an agent capable of blocking EHEC-induced increases in epithelial permeability via maintenance of claudin-2, occludin, and zonula occludens-1 levels. PMID- 16314473 TI - Potential pathogenetic implications of cyclooxygenase-2 overexpression in B chronic lymphoid leukemia cells. AB - Evidence suggests that cyclooxygenase-2 (COX-2) increases tumorigenic potential by promoting resistance to apoptosis. Because B chronic lymphoid leukemia (B-CLL) cells exhibit a defective apoptotic response, we analyzed CD19(+) B lymphocytes purified from the peripheral blood of B-CLL patients. Microarray analysis showed a variable (up to 38-fold) increase in the steady-state mRNA levels of COX-2 in B CLL lymphocytes compared with normal CD19(+) B lymphocytes. The up-regulation of COX-2 in B-CLL cells was confirmed by reverse transcriptase-polymerase chain reaction and Western blot analyses. Moreover, immunohistochemical analysis of B CLL bone marrow infiltrates confirmed clear expression of COX-2 in leukemic cells. Ex vivo treatment with the COX-2 inhibitor NS-398 significantly decreased the survival of leukemic cells by increasing the rate of spontaneous apoptosis in 13 of 16 B-CLL samples examined, but it did not affect the survival of normal lymphocytes. Pretreatment with NS-398 significantly potentiated the cytotoxicity induced by chlorambucil in 8 of 16 B-CLL samples examined. Moreover, although recombinant tumor necrosis factor-related apoptosis inducing ligand (TRAIL)/Apo2L showed little cytotoxic effect in most B-CLL samples examined, pretreatment with NS-398 sensitized 8 of 16 B-CLL samples to TRAIL-induced apoptosis. Taken together, our data indicate that COX-2 overexpression likely represents an additional mechanism of resistance to apoptosis in B-CLL and that pharmacological suppression of COX-2 might enhance chemotherapy-mediated apoptosis. PMID- 16314474 TI - Muscle wasting induced by HTLV-1 tax-1 protein: an in vitro and in vivo study. AB - Besides tropical spastic paraparesis/human T-cell leukemia virus type-1 (HTLV-1) associated myelopathy, the human retrovirus HTLV-1 causes inflammatory disorders such as myositis. Although the pathogenesis of HTLV-1-associated myositis is primarily unknown, a direct effect of cytokines or viral proteins in myocytotoxicity is suspected. We have developed an in vitro cell culture model to study the interactions between primary human muscle cells and HTLV-1 chronically infected cells. When HTLV-1-infected cell lines were added to differentiated muscle cultures, cytopathic changes such as fiber shrinking were observed as early as 1 day after contact. This was accompanied by alterations in desmin and vimentin organization, occurring in the absence of muscle cell infection but with Tax-1 present in myotubes. Cytopathic changes were also observed when infected culture supernatants were added to the muscle cells. Fiber atrophy and cytoskeletal disorganization were confirmed in muscle biopsies from two HTLV-1 infected patients with myositis. Transduction of cultured muscle cells with a lentiviral vector containing the HTLV-1 Tax gene reproduced such effects in vitro. The present data indicate that the myocytotoxicity that is observed in HTLV-1-associated myopathies can be due to a direct effect of the Tax-1 protein expressed in infected inflammatory cells, in the absence of muscle cell infection. PMID- 16314475 TI - Oral and nasal sensitization promote distinct immune responses and lung reactivity in a mouse model of peanut allergy. AB - Despite structural and functional differences between the initial sites of contact with allergens in the gastrointestinal and nasal tracts, few animal models have examined the influence of the mucosal routes of sensitization on host reactivity to food or environmental antigens. We compared the oral and nasal routes of peanut sensitization for the development of a mouse model of allergy. Mice were sensitized by administration of peanut proteins in the presence of cholera toxin as adjuvant. Antibody and cytokine responses were characterized, as well as airway reactivity to nasal challenge with peanut or unrelated antigens. Oral sensitization promoted higher levels of IgE, but lower IgG responses, than nasal sensitization. Both orally and nasally sensitized mice experienced airway hyperreactivity on nasal peanut challenge. The peanut challenge also induced lung eosinophilia and type 2 helper T-cell-type cytokines in orally sensitized mice. In contrast, peanut challenge in nasally sensitized mice promoted neutrophilia and higher levels of lung MAC-1(+) I-A(b low) cells and inflammatory cytokines. In addition, nasal but not oral, sensitization promoted lung inflammatory responses to unrelated antigens. In summary, both oral and nasal peanut sensitization prime mice for airway hyperreactivity, but the initial mucosal route of sensitization influences the nature of lung inflammatory responses to peanut and unrelated allergens. PMID- 16314476 TI - Massive apoptosis in lymphoid organs in animal models for primary and secondary progressive multiple sclerosis. AB - The mechanism(s) responsible for generating the different forms of multiple sclerosis, primary progressive (PP) and secondary progressive (SP) versus relapsing-remitting (RR), is not well understood. Using myelin oligodendrocyte glycoprotein (MOG)(92-106), we have established animal models that mimic the different types of multiple sclerosis. A.SW mice develop PP or SP-experimental allergic encephalomyelitis (EAE) with large areas of demyelination and high titers of MOG antibody whereas SJL/J mice develop RR-EAE with perivascular T cells and mild demyelination. In A.SW progressive EAE, we found atrophy of the thymus, spleen, and lymph nodes with depletion of T and B cells and massive apoptosis, as demonstrated by immunohistochemistry, terminal dUTP nick-end labeling, and DNA agarose gel electrophoresis. To test whether lymphoid apoptosis itself contributes to disease progression, we injected SJL/J mice with apoptotic thymocytes. Injection of apoptotic cells resulted in greater than 20% of mice developing SP-EAE with ataxia. SJL/J mice with SP-EAE had large areas of demyelination, high MOG antibody titers and atrophic lymphoid organs. Spleen cells from mice with progressive EAE produced less interferon-gamma than those from RR-EAE when stimulated with mitogen. We suggest that induction of lymphoid apoptosis alters the balance of Th1 versus Th2 immune responses and increases MOG antibody production, leading to exacerbation of demyelination and subsequent disease progression. PMID- 16314477 TI - P-selectin can support both Th1 and Th2 lymphocyte rolling in the intestinal microvasculature. AB - Lymphocyte localization to inflammatory sites is paramount for developing and maintaining an immune response. Rolling is the first step in recruitment, but our knowledge of its mechanisms in Th1 and Th2 CD4(+) lymphocytes is incomplete. Whereas initial studies suggested that Th1 but not Th2 lymphocytes used P selectin for recruitment, more recent studies have proposed that both subtypes bind selectins. We used intravital microscopy to demonstrate in vivo that polarized Th1 and Th2 lymphocytes both use P-selectin to roll and adhere to cytokine [tumor necrosis factor (TNF)-alpha or interleukin (IL)-4]-activated intestinal microvasculature. The majority of Th1 lymphocyte flux in TNF-alpha- and IL-4-treated animals was P-selectin-dependent. Th1 lymphocytes also interacted with E-selectin to control rolling velocity after TNF-alpha stimulation. Th2 lymphocytes, which make IL-4 but not interferon-gamma, bound P selectin ex vivo, with more than 95% rolling on P-selectin in vivo. Both Th1 and Th2 lymphocytes regulated rolling velocity by interacting with alpha(4)-integrin. Furthermore, in a model of spontaneous intestinal inflammation (ie, IL-10 deficient mice), both Th1 and Th2 lymphocytes rolled, adhered, and ultimately emigrated into the local microenvironment. These results suggest that both Th1 and Th2 lymphocytes use P-selectin in the initial rolling step in vivo in response to a global activator of the vasculature (TNF), a subtle inducer of P selectin (IL-4), and pathological inflammation (IL-10-deficient mice). PMID- 16314478 TI - Both Th1 and Th2 cells require P-selectin glycoprotein ligand-1 for optimal rolling on inflamed endothelium. AB - The acquisition of homing receptors that redirect lymphocyte trafficking to nonlymphoid tissues after antigen encounter is a fundamental aspect of effector T cell development. Although a role for selectins and their ligands has been well characterized for trafficking of Th1 cells to nonlymphoid sites, mechanisms responsible for Th2 trafficking are not well understood. Using a flow chamber system in which the endothelial interactions of two distinct T-cell populations could be examined simultaneously, we directly compared the requirements for Th1 and Th2 cell tethering and rolling. We found that although Th2 cells expressed significantly lower levels of selectin ligands than Th1 cells, activation of the endothelium by Th2-derived factors induced rolling interactions that were comparable for both Th1 and Th2 populations. Further, in the absence of PSGL-1, no other adhesion molecule could effectively compensate for lack of PSGL-1 to mediate rolling of either Th1 or Th2 cells. Thus, both Th1 and Th2 populations express functional PSGL-1-based selectin ligands for tethering and rolling on activated endothelium, and both effector populations can use PSGL-1 as the dominant scaffold for functional selectin ligand expression. PMID- 16314479 TI - Membrane tumor necrosis factor confers partial protection to Listeria infection. AB - Tumor necrosis factor (TNF) plays a critical role in the host response to the intracellular pathogen Listeria monocytogenes (LM). TNF exists in soluble and membrane-bound forms and exhibits both unique and overlapping activities. We examined the role of membrane TNF in the absence of secreted TNF for host resistance in knockin mice in which the endogenous TNF was replaced by a regulated, noncleavable allele (mem-TNF). Macrophages expressing mem-TNF produced nitric oxide and displayed normal bactericidal activity. Although mice completely deficient in TNF (TNF(-/-)) succumbed to LM infection within 4 days, mem-TNF mice controlled LM infection at a low dose (10(4) CFU) but succumbed at a higher dose of infection (10(5) CFU). In contrast to complete TNF deficiency, mem-TNF mice developed confined microabscesses that expressed inducible nitric oxide synthase. The transfer of lymphocytes from immunized mem-TNF, but not TNF(-/-), mice protected TNF(-/-) mice from fatal infection. Taken together the data suggest that in the absence of soluble TNF, the presence of membrane-expressed TNF on phagocytes and lymphocytes partially restores host defense to LM infection. PMID- 16314480 TI - Primary and secondary elastin-binding protein defect leads to impaired elastogenesis in fibroblasts from GM1-gangliosidosis patients. AB - G(M1)-gangliosidosis is a lysosomal storage disorder caused by acid beta galactosidase deficiency. Aside from the lysosomal beta-galactosidase enzyme, the beta-galactosidase gene also encodes the elastin-binding protein (EBP), deficiency in which impairs elastogenesis. Using expression studies and Western blots of COS-1 cells, we identified and characterized four new and two known beta galactosidase gene mutations detected in G(M1)-gangliosidosis patients with infantile, juvenile, or adult forms of disease. We then focused on impaired elastogenesis detected in fibroblasts from patients with infantile and juvenile disease. The juvenile patient showed connective-tissue abnormalities, unusual urinary keratan sulfate excretion, and an EBP reduction, despite mutations affecting only beta-galactosidase. Because galactosugar-bearing moieties may alter EBP function and impair elastogenesis, we assessed infantile and juvenile patients for the source of altered elastogenesis. We confirmed that the infantile patient's impaired elastogenesis arose from a primary EBP defect, according to molecular analysis. We examined the juvenile's fibroblasts by immunohistochemistry, addition of keratanase, soluble/insoluble elastin assay, and radiolabeling of tropoelastin. These experiments revealed that the juvenile's impaired elastogenesis likely arose from secondary EBP deficiency caused by keratan sulfate accumulation. Thus, impaired elastogenesis in G(M1) gangliosidosis can arise from primary or secondary EBP defects in fibroblasts from infantile and juvenile patients, respectively. PMID- 16314481 TI - Matrix contraction by dermal fibroblasts requires transforming growth factor beta/activin-linked kinase 5, heparan sulfate-containing proteoglycans, and MEK/ERK: insights into pathological scarring in chronic fibrotic disease. AB - Scarring is characterized by excessive synthesis and contraction of extracellular matrix. Here, we show that fibroblasts from scarred (lesional) areas of patients with the chronic fibrotic disorder diffuse scleroderma [diffuse systemic sclerosis (dSSc)] show an enhanced ability to adhere to and contract extracellular matrix, relative to fibroblasts from unscarred (nonlesional) areas of dSSc patients and dermal fibroblasts from normal, healthy individuals. The contractile abilities of normal and dSSc dermal fibroblasts were suppressed by blocking heparin sulfate-containing proteoglycan biosynthesis or antagonizing transforming growth factor-beta receptor type I [activin-linked kinase (ALK5)] or ras/mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinase (ERK). Compared with both normal and nonlesional fibroblasts, lesional dSSc fibroblasts overexpressed the heparin sulfate-containing proteoglycan syndecan 4. We also found that the procontractile signals from transforming growth factor (TGF)-beta were integrated through syndecan 4 and MEK/ERK because the ability of TGFbeta to induce contraction of dermal fibroblasts was prevented by MEK antagonism. TGFbeta could not induce a contractile phenotype or phosphorylate ERK in syndecan 4(-/-) dermal fibroblasts. These results suggest that integrating TGFbeta and ERK signals via syndecan 4 is essential for the contractile ability of dermal fibroblasts. We conclude that antagonizing MEK/ERK, TGFbeta1/ALK5, or syndecan 4 may alleviate scarring in chronic fibrotic disease. PMID- 16314482 TI - Participation of autophagy in storage of lysosomes in neurons from mouse models of neuronal ceroid-lipofuscinoses (Batten disease). AB - In cathepsin D-deficient (CD-/-) and cathepsins B and L double-deficient (CB-/-CL /-) mice, abnormal vacuolar structures accumulate in neurons of the brains. Many of these structures resemble autophagosomes in which part of the cytoplasm is retained but their precise nature and biogenesis remain unknown. We show here how autophagy contributes to the accumulation of these vacuolar structures in neurons deficient in cathepsin D or both cathepsins B and L by demonstrating an increased conversion of the molecular form of MAP1-LC3 for autophagosome formation from the cytosolic form (LC3-I) to the membrane-bound form (LC3-II). In both CD-/- and CB /-CL-/- mouse brains, the membrane-bound LC3-II form predominated whereas MAP1 LC3 signals accumulated in granular structures located in neuronal perikarya and axons of these mutant brains and were localized to the membranes of autophagosomes, evidenced by immunofluorescence microscopy and freeze-fracture replica immunoelectron microscopy. Moreover, as in CD-/- neurons, autofluorescence and subunit c of mitochondrial ATP synthase accumulated in CB-/ CL-/- neurons. This suggests that not only CD-/- but also CB-/-CL-/- mice could be useful animal models for neuronal ceroid-lipofuscinosis/Batten disease. These data strongly argue for a major involvement of autophagy in the pathogenesis of Batten disease/lysosomal storage disorders. PMID- 16314483 TI - Creutzfeldt-Jakob disease (CJD) with a mutation at codon 148 of prion protein gene: relationship with sporadic CJD. AB - Creutzfeldt-Jakob disease (CJD), the most common human prion disease, includes sporadic (s) and familial (f) forms. Regardless of etiology, both forms are thought to share the pathogenic mechanism whereby the cellular prion protein (PrP(C)) converts into its pathogenic isoform (PrP(Sc)). While PrP(C) conversion is thought to be random in sCJD, conversion in fCJD is facilitated by the congenital presence of mutated PrP. Differences in PrP genotype (PRNP) and in conversion circumstances lead to PrP(Sc) with distinct characteristics that elicit different disease phenotypes. Here, we describe a case of fCJD with a substitution of histidine (H) for arginine (R) at codon 148 (R148H) and heterozygosity of the methionine/valine (M/V) polymorphic codon 129, with the 129M allele coupled with the mutation. The disease phenotype and all major characteristics of PrP(Sc) of fCJD(R148H) were virtually indistinguishable from those of sCJDMV2, which has features different from those of any other sCJD. Therefore, despite the differences in etiology, PRNP, and conversion process, the two forms of PrP(Sc) had similar characteristics. Furthermore, comparison of fCJD(R148H) with a recently reported case carrying R148H and homozygosity at codon 129 suggests that codon 129 coupled with the mutation as well as that located on the normal allele can modify major phenotypic and PrP(Sc) features of fCJD(R148H). PMID- 16314484 TI - Absence of host-secreted protein acidic and rich in cysteine (SPARC) augments peritoneal ovarian carcinomatosis. AB - The matricellular glycoprotein SPARC (secreted protein acidic and rich in cysteine) possesses multifaceted roles in modulation of cell-matrix interactions, as well as tumor growth and metastasis. To investigate the influence of host derived SPARC on peritoneal dissemination of ovarian cancer, we established a murine model that faithfully recapitulates advanced human disease by intraperitoneal injection of syngeneic ID8 ovarian cancer cells into SPARC-null and wild-type mice. Compared to wild-type mice, SPARC-null mice showed significantly shorter survival and developed extensive nodular peritoneal dissemination with hemorrhagic ascitic fluid accumulation. Ascitic fluid collected from SPARC-null mice showed significantly augmented levels and activity of vascular endothelial growth factor and gelatinases. Immunohistochemical analysis of tumor nodules from SPARC-null mice revealed higher proliferation and lower apoptosis indices with minimal staining for major extracellular matrix constituents. In vitro, SPARC significantly suppressed adhesion to and invasion of various peritoneal extracellular matrix constituents by murine and human ovarian cancer cell lines. Our findings suggest that SPARC ameliorates ovarian peritoneal carcinomatosis through abrogation of the initial steps of disease pathogenesis, namely tumor cell adhesion and invasion, inhibition of tumor cell proliferation, and induction of apoptosis. Thus, SPARC represents an important therapeutic candidate in ovarian cancer. PMID- 16314485 TI - Heterogeneity of Tie2 expression in tumor microcirculation: influence of cancer type, implantation site, and response to therapy. AB - To evaluate the expression of the Tie2/Tek tyrosine kinase receptor in tumor blood vessels, we examined Tie2lacZ(+)/RAG1(-) mice. There was considerable heterogeneity (Tie2-negative, Tie2-positive, or Tie2-composite blood vessels) in subcutaneous xenografts of human colorectal carcinoma (HCT116; 97.5% Tie2 positive vessels) versus human melanoma (WM115; 75.9% Tie2-positive vessels). Similar patterns of Tie2 expression occurred in abdominal metastases derived from the same cell lines. Immunostaining for endothelial markers and Tie2 revealed that endogenous protein levels corresponded with transgene activity. Endothelial cells were confirmed to be of mouse origin through triple immunofluorescence staining with mouse antiserum to human nuclei, isolectin GS-IB(4), and anti-Tie2. Similar Tie2 heterogeneity was observed in clinical specimens from a variety of human cancers, including malignant melanoma and colorectal carcinoma. We also examined the effect of Tek-Delta Fc anti-angiogenic therapy on tumor growth and Tie2 expression patterns in HCT116 and WM115 subcutaneous xenografts. Tek-Delta induced extensive tumor regression in HCT116 tumors and concomitant reductions in Tie2-expressing blood vessels. However, no significant responses were seen in Tek Delta-treated WM115 tumors. Thus, vascular heterogeneity of Tie2 expression is cancer-type specific, suggesting that the tumor microenvironment and/or direct cancer cell interactions influence Tie2 endothelial expression. PMID- 16314486 TI - Analysis of orthologous gene expression between human pulmonary adenocarcinoma and a carcinogen-induced murine model. AB - Human adenocarcinoma (AC) is the most frequently diagnosed human lung cancer, and its absolute incidence is increasing dramatically. Compared to human lung AC, the A/J mouse-urethane model exhibits similar histological appearance and molecular changes. We examined the gene expression profiles of human and murine lung tissues (normal or AC) and compared the two species' datasets after aligning approximately 7500 orthologous genes. A list of 409 gene classifiers (P value <0.0001), common to both species (joint classifiers), showed significant, positive correlation in expression levels between the two species. A number of previously reported expression changes were recapitulated in both species, such as changes in glycolytic enzymes and cell-cycle proteins. Unexpectedly, joint classifiers in angiogenesis were uniformly down-regulated in tumor tissues. The eicosanoid pathway enzymes prostacyclin synthase (PGIS) and inducible prostaglandin E(2) synthase (PGES) were joint classifiers that showed opposite effects in lung AC (PGIS down-regulated; PGES up-regulated). Finally, tissue microarrays identified the same protein expression pattern for PGIS and PGES in 108 different non-small cell lung cancer biopsies, and the detection of PGIS had statistically significant prognostic value in patient survival. Thus, the A/J mouse-urethane model reflects significant molecular details of human lung AC, and comparison of changes in orthologous gene expression may provide novel insights into lung carcinogenesis. PMID- 16314487 TI - LIF and BMP signaling generate separate and discrete types of GFAP-expressing cells. AB - Bone morphogenetic protein (BMP) and leukemia inhibitory factor (LIF) signaling both promote the differentiation of neural stem/progenitor cells into glial fibrillary acidic protein (GFAP) immunoreactive cells. This study compares the cellular and molecular characteristics, and the potentiality, of GFAP(+) cells generated by these different signaling pathways. Treatment of cultured embryonic subventricular zone (SVZ) progenitor cells with LIF generates GFAP(+) cells that have a bipolar/tripolar morphology, remain in cell cycle, contain progenitor cell markers and demonstrate self-renewal with enhanced neurogenesis - characteristics that are typical of adult SVZ and subgranular zone (SGZ) stem cells/astrocytes. By contrast, BMP-induced GFAP(+) cells are stellate, exit the cell cycle, and lack progenitor traits and self-renewal--characteristics that are typical of astrocytes in the non-neurogenic adult cortex. In vivo, transgenic overexpression of BMP4 increases the number of GFAP(+) astrocytes but depletes the GFAP(+) progenitor cell pool, whereas transgenic inhibition of BMP signaling increases the size of the GFAP(+) progenitor cell pool but reduces the overall numbers of astrocytes. We conclude that LIF and BMP signaling generate different astrocytic cell types, and propose that these cells are, respectively, adult progenitor cells and mature astrocytes. PMID- 16314488 TI - The dissociation of the Fgf-feedback loop controls the limbless state of the neck. AB - In tetrapods, limbs develop at two specific positions along the anteroposterior axis of the embryo, whereas other regions of the embryo, most prominently the neck and the flank, are limbless. However, the flank can generate an ectopic limb when the Fgf-feedback loop crucial for the initiation of limb budding is activated. Thus, despite its limblessness, the flank is a limb-competent area. Using the chick embryo as model, we investigated whether the neck, as the flank, has the competence to form a limb, and what mechanism may regulate its limblessness. We show that forelimb lateral mesoderm plus ectoderm grafted into the neck can continue limb development, suggesting that the neck does not actively inhibit this process. However, neck tissues themselves do not support or take part in limb formation. Hence, the neck is limb-incompetent. This is due to the dismantling of Fgf signalling at distinct points of the MAPK signalling cascade in the neck lateral mesoderm and ectoderm. PMID- 16314489 TI - Inactivation of NF1 in CNS causes increased glial progenitor proliferation and optic glioma formation. AB - The gene responsible for neurofibromatosis type 1 (NF1) encodes a tumor suppressor that functions as a negative regulator of the Ras proto-oncogene. Individuals with germline mutations in NF1 are predisposed to the development of benign and malignant tumors of the peripheral and central nervous system (CNS). Children with this disease suffer a high incidence of optic gliomas, a benign but potentially debilitating tumor of the optic nerve; and an increased incidence of malignant astrocytoma, reactive astrogliosis and intellectual deficits. In the present study, we have sought insight into the molecular and cellular basis of NF1-associated CNS pathologies. We show that mice genetically engineered to lack NF1 in CNS exhibit a variety of defects in glial cells. Primary among these is a developmental defect resulting in global reactive astrogliosis in the adult brain and increased proliferation of glial progenitor cells leading to enlarged optic nerves. As a consequence, all of the mutant optic nerves develop hyperplastic lesions, some of which progress to optic pathway gliomas. These data point to hyperproliferative glial progenitors as the source of the optic tumors and provide a genetic model for NF1-associated astrogliosis and optic glioma. PMID- 16314490 TI - Molecular characterization of melanocyte stem cells in their niche. AB - Emerging evidence from stem cell (SC) research has strengthened the idea that SC fate is determined by a specialized environment, known as the SC niche. However, because of the difficulty of identifying individual stem cells and their surrounding components in situ, the exact mechanisms underlying SC regulation by the niche remain elusive. To overcome this difficulty, we employed melanocyte stem cells (MSCs), which allow the identification of individual SCs in the niche, the lower permanent portion of the hair follicle (HF). Here, we present molecular makers that can distinguish MSCs from other melanocyte (MC) subsets in the HF. We also describe a simple and robust method that allows gene expression profiling in individual SCs. After isolating individual MSCs from transgenic mice in which the MCs are marked by green fluorescence protein (GFP), we performed single-cell transcript analysis to obtain the molecular signature of individual MSCs in the niche. The data suggest the existence of a mechanism that induces the downregulation of various key molecules for MC proliferation or differentiation in MSCs located in the niche. By integrating these data, we propose that the niche is an environment that insulates SCs from various activating stimuli and maintains them in a quiescent state. PMID- 16314492 TI - Retraction: Dose dependency of Disp1 and genetic interaction between Disp1 and other hedgehog signaling components in the mouse. PMID- 16314491 TI - Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning. AB - Cardiac cushion development provides a valuable system to investigate epithelial to mesenchymal transition (EMT), a fundamental process in development and tumor progression. In the atrioventricular (AV) canal, endocardial cells lining the heart respond to a myocardial-derived signal, undergo EMT, and contribute to cushion mesenchyme. Here, we inactivated bone morphogenetic protein 2 (Bmp2) in the AV myocardium of mice. We show that Bmp2 has three functions in the AV canal: to enhance formation of the cardiac jelly, to induce endocardial EMT and to pattern the AV myocardium. Bmp2 is required for myocardial expression of Has2, a crucial component of the cardiac jelly matrix. During EMT, Bmp2 promotes expression of the basic helix-loop-helix factor Twist1, previously implicated in EMT in cancer metastases, and the homeobox genes Msx1 and Msx2. Deletion of the Bmp type 1A receptor, Bmpr1a, in endocardium also resulted in failed cushion formation, indicating that Bmp2 signals directly to cushion-forming endocardium to induce EMT. Lastly, we show that Bmp2 mutants failed to specify the AV myocardium with loss of Tbx2 expression uncovering a myocardial, planar signaling function for Bmp2. Our data indicate that Bmp2 has a crucial role in coordinating multiple aspects of AV canal morphogenesis. PMID- 16314493 TI - Obstructive sleep apnea in overweight subjects. PMID- 16314494 TI - Runx1 binds positive transcription elongation factor b and represses transcriptional elongation by RNA polymerase II: possible mechanism of CD4 silencing. AB - Runx1 binds the silencer and represses CD4 transcription in immature thymocytes. In this study, we found that Runx1 inhibits P-TEFb, which contains CycT1, CycT2, or CycK and Cdk9 and stimulates transcriptional elongation by RNA polymerase II (RNAPII) in eukaryotic cells. Indeed, its inhibitory domain, spanning positions 371 to 411, not only bound CycT1 but was required for silencing CD4 transcription in vivo. Our chromatin immunoprecipitation assays revealed that Runx1 inhibits the elongation but not initiation of transcription and that RNAPII is engaged at the CD4 promoter but is unable to elongate in CD4(-) CD8(+) thymoma cells. These results suggest that active repression by Runx1 occurs by blocking the elongation by RNAPII, which may contribute to CD4 silencing during T-cell development. PMID- 16314495 TI - PGC-1alpha coactivates PDK4 gene expression via the orphan nuclear receptor ERRalpha: a mechanism for transcriptional control of muscle glucose metabolism. AB - The transcriptional coactivator PGC-1alpha is a key regulator of energy metabolism, yet little is known about its role in control of substrate selection. We found that physiological stimuli known to induce PGC-1alpha expression in skeletal muscle coordinately upregulate the expression of pyruvate dehydrogenase kinase 4 (PDK4), a negative regulator of glucose oxidation. Forced expression of PGC-1alpha in C(2)C(12) myotubes induced PDK4 mRNA and protein expression. PGC 1alpha-mediated activation of PDK4 expression was shown to occur at the transcriptional level and was mapped to a putative nuclear receptor binding site. Gel shift assays demonstrated that the PGC-1alpha-responsive element bound the estrogen-related receptor alpha (ERRalpha), a recently identified component of the PGC-1alpha signaling pathway. In addition, PGC-1alpha was shown to activate ERRalpha expression. Chromatin immunoprecipitation assays confirmed that PGC 1alpha and ERRalpha occupied the mPDK4 promoter in C(2)C(12) myotubes. Additionally, transfection studies using ERRalpha-null primary fibroblasts demonstrated that ERRalpha is required for PGC-1alpha-mediated activation of the mPDK4 promoter. As predicted by the effects of PGC-1alpha on PDK4 gene transcription, overexpression of PGC-1alpha in C(2)C(12) myotubes decreased glucose oxidation rates. These results identify the PDK4 gene as a new PGC 1alpha/ERRalpha target and suggest a mechanism whereby PGC-1alpha exerts reciprocal inhibitory influences on glucose catabolism while increasing alternate mitochondrial oxidative pathways in skeletal muscle. PMID- 16314496 TI - Activation of TRAP/mediator subunit TRAP220/Med1 is regulated by mitogen activated protein kinase-dependent phosphorylation. AB - The TRAP/Mediator coactivator complex serves as a molecular bridge between gene specific activators and RNA polymerase II. TRAP220/Med1 is a key component of TRAP/Mediator that targets the complex to nuclear hormone receptors and other types of activators. We show here that human TRAP220/Med1 is a specific substrate for extracellular signal-regulated kinase (ERK) of the mitogen-activated protein kinase (MAPK) family. We demonstrate that ERK phosphorylates TRAP220/Med1 in vivo at two specific sites: threonine 1032 and threonine 1457. Importantly, we found that ERK phosphorylation significantly increases the stability and half-life of TRAP220/Med1 in vivo and correlates with increased thyroid hormone receptor dependent transcription. Furthermore, ERK phosphorylates TRAP220/Med1 in a cell cycle-dependent manner, resulting in peak levels of expression during the G(2)/M phase of the cell cycle. ERK phosphorylation of ectopic TRAP220/Med1 also triggered shuttling into the nucleolus, thus suggesting that ERK may regulate TRAP220/Med1 subnuclear localization. Finally, we observed that ERK phosphorylation of TRAP220/Med1 stimulates its intrinsic transcriptional coactivation activity. We propose that ERK-mediated phosphorylation is a regulatory mechanism that controls TRAP220/Med1 expression levels and modulates its functional activity. PMID- 16314497 TI - An eh1-like motif in odd-skipped mediates recruitment of Groucho and repression in vivo. AB - Drosophila Groucho, like its vertebrate Transducin-like Enhancer-of-split homologues, is a corepressor that silences gene expression in numerous developmental settings. Groucho itself does not bind DNA but is recruited to target promoters by associating with a large number of DNA-binding negative transcriptional regulators. These repressors tether Groucho via short conserved polypeptide sequences, of which two have been defined. First, WRPW and related tetrapeptide motifs have been well characterized in several repressors. Second, a motif termed Engrailed homology 1 (eh1) has been found predominantly in homeodomain-containing transcription factors. Here we describe a yeast two-hybrid screen that uncovered physical interactions between Groucho and transcription factors, containing eh1 motifs, with different types of DNA-binding domains. We show that one of these, the zinc finger protein Odd-skipped, requires its eh1 like sequence for repressing specific target genes in segmentation. Comparison between diverse eh1 motifs reveals a bias for the phosphoacceptor amino acids serine and threonine at a fixed position, and a mutational analysis of Odd skipped indicates that these residues are critical for efficient interactions with Groucho and for repression in vivo. Our data suggest that phosphorylation of these phosphomeric residues, if it occurs, will down-regulate Groucho binding and therefore repression, providing a mechanism for posttranslational control of Groucho-mediated repression. PMID- 16314498 TI - Cooperative control of Crb2 by ATM family and Cdc2 kinases is essential for the DNA damage checkpoint in fission yeast. AB - The cellular responses to double-stranded breaks (DSBs) typically involve the extensive accumulation of checkpoint proteins in chromatin surrounding the damaged DNA. One well-characterized example involves the checkpoint protein Crb2 in the fission yeast Schizosaccharomyces pombe. The accumulation of Crb2 at DSBs requires the C-terminal phosphorylation of histone H2A (known as gamma-H2A) by ATM family kinases in chromatin surrounding the break. It also requires the constitutive methylation of histone H4 on lysine-20 (K20). Interestingly, neither type of histone modification is essential for the Crb2-dependent checkpoint response. However, H4-K20 methylation is essential in a crb2-T215A strain that lacks a cyclin-dependent kinase phosphorylation site in Crb2. Here we explain this genetic interaction by describing a previously overlooked effect of the crb2 T215A mutation. We show that crb2-T215A cells are able to initiate but not sustain a checkpoint response. We also report that gamma-H2A is essential for the DNA damage checkpoint in crb2-T215A cells. Importantly, we show that inactivation of Cdc2 in gamma-H2A-defective cells impairs Crb2-dependent signaling to the checkpoint kinase Chk1. These findings demonstrate that full Crb2 activity requires phosphorylation of threonine-215 by Cdc2. This regulation of Crb2 is independent of the histone modifications that are required for the hyperaccumulation of Crb2 at DSBs. PMID- 16314499 TI - Requirement for the SnoN oncoprotein in transforming growth factor beta-induced oncogenic transformation of fibroblast cells. AB - Transforming growth factor beta (TGF-beta) was originally identified by virtue of its ability to induce transformation of the AKR-2B and NRK fibroblasts but was later found to be a potent inhibitor of the growth of epithelial, endothelial, and lymphoid cells. Although the growth-inhibitory pathway of TGF-beta mediated by the Smad proteins is well studied, the signaling pathway leading to the transforming activity of TGF-beta in fibroblasts is not well understood. Here we show that SnoN, a member of the Ski family of oncoproteins, is required for TGF beta-induced proliferation and transformation of AKR-2B and NRK fibroblasts. TGF beta induces upregulation of snoN expression in both epithelial cells and fibroblasts through a common Smad-dependent mechanism. However, a strong and prolonged activation of snoN transcription that lasts for 8 to 24 h is detected only in these two fibroblast lines. This prolonged induction is mediated by Smad2 and appears to play an important role in the transformation of both AKR-2B and NRK cells. Reduction of snoN expression by small interfering RNA or shortening of the duration of snoN induction by a pharmacological inhibitor impaired TGF-beta induced anchorage-independent growth of AKR-2B cells. Interestingly, Smad2 and Smad3 play opposite roles in regulating snoN expression in both fibroblasts and epithelial cells. The Smad2/Smad4 complex activates snoN transcription by direct binding to the TGF-beta-responsive element in the snoN promoter, while the Smad3/Smad4 complex inhibits it through a novel Smad inhibitory site. Mutations of Smad4 that render it defective in heterodimerization with Smad3, which are found in many human cancers, convert the activity of Smad3 on the snoN promoter from inhibitory to stimulatory, resulting in increased snoN expression in cancer cells. Thus, we demonstrate a novel role of SnoN in the transforming activity of TGF-beta in fibroblasts and also uncovered a mechanism for the elevated SnoN expression in some human cancer cells. PMID- 16314500 TI - Interactions of the yeast SF3b splicing factor. AB - The U2 snRNP promotes prespliceosome assembly through interactions that minimally involve the branchpoint binding protein, Mud2p, and the pre-mRNA. We previously showed that seven proteins copurify with the yeast (Saccharomyces cerevisiae) SF3b U2 subcomplex that associates with the pre-mRNA branchpoint region: Rse1p, Hsh155p, Hsh49p, Cus1p, and Rds3p and unidentified subunits p10 and p17. Here proteomic and genetic studies identify Rcp10p as p10 and show that it contributes to SF3b stability and is necessary for normal cellular Cus1p accumulation and for U2 snRNP recruitment in splicing. Remarkably, only the final 53 amino acids of Rcp10p are essential. p17 is shown to be composed of two accessory splicing factors, Bud31p and Ist3p, the latter of which independently associates with the RES complex implicated in the nuclear pre-mRNA retention. A directed two-hybrid screen reveals a network of prospective interactions that includes previously unreported intra-SF3b contacts and SF3b interactions with the RES subunit Bud13p, the Prp5p DExD/H-box protein, Mud2p, and the late-acting nineteen complex. These data establish the concordance of yeast and mammalian SF3b complexes, implicate accessory splicing factors in U2 snRNP function, and support SF3b contribution from early pre-mRNP recognition to late steps in splicing. PMID- 16314501 TI - Nucleosome stability at the yeast PHO5 and PHO8 promoters correlates with differential cofactor requirements for chromatin opening. AB - The coregulated PHO5 and PHO8 genes in Saccharomyces cerevisiae provide typical examples for the role of chromatin in promoter regulation. It has been a long standing question why the cofactors Snf2 and Gcn5 are essential for full induction of PHO8 but dispensable for opening of the PHO5 promoter. We show that this discrepancy may result from different stabilities of the two promoter chromatin structures. To test this hypothesis, we used our recently established yeast extract in vitro chromatin assembly system, which generates the characteristic PHO5 promoter chromatin. Here we show that this system also assembles the native PHO8 promoter nucleosome pattern. Remarkably, the positioning information for both native patterns is specific to the yeast extract. Salt gradient dialysis or Drosophila embryo extract does not support proper nucleosome positioning unless supplemented with yeast extract. By competitive assemblies in the yeast extract system we show that the PHO8 promoter has greater nucleosome positioning power and that the properly positioned nucleosomes are more stable than those at the PHO5 promoter. Thus we provide evidence for the correlation of inherently more stable chromatin with stricter cofactor requirements. PMID- 16314502 TI - Involvement of Prep1 in the alphabeta T-cell receptor T-lymphocytic potential of hematopoietic precursors. AB - Prep1 is a homeodomain transcription factor that acts by dimerizing with Pbx. Since Prep1 null embryos die at gastrulation, we studied Prep1(i/i) hypomorphic mice to study the physiological role of Prep1. A low percentage of homozygous Prep1(i/i) mice survived at birth, and their postnatal functions could be investigated. Reduced Prep1 expression caused an abnormal thymic T-cell development: increased CD4(-) CD8(-) double-negative thymocytes, decrease in alphabetaTCR(high) cells (cells with high levels of the alphabetaTau-cell receptor [alphabetaTCR]) and CD4(+) and CD8(+) single-positive (SP) thymocytes, and increase in gammadeltaTCR cells. Peripheral lymphoid organs of Prep1(i/i) mice contained fewer alphabetaTCR mature T cells and more gammadeltaTCR T cells than wild-type littermates. Moreover, Prep1(i/i) CD4(+) CD8(+) double-positive thymocytes underwent more apoptosis, and SP thymocytes proliferated less than control littermates. Mice that were lethally irradiated and then had Prep1(i/i) fetal liver cells transplanted showed the same defects as the Prep1(i/i) mice did. Among PBC family members, Pbx2 and very low levels of Pbx3 were observed in the thymi of wild-type mice. In Prep1(i/i) mice, the level of Pbx2 protein was profoundly decreased, while for Pbx3 no definitive conclusion could be reached. Therefore, the deficient postnatal T-lymphocytic potential of the Prep1 hematopoietic progenitors depends on the combined, not compensated, absence of Prep1 and at least Pbx2. PMID- 16314503 TI - Mutations of the Yku80 C terminus and Xrs2 FHA domain specifically block yeast nonhomologous end joining. AB - The nonhomologous end-joining (NHEJ) pathway of DNA double-strand break repair requires three protein complexes in Saccharomyces cerevisiae: MRX (Mre11-Rad50 Xrs2), Ku (Ku70-Ku80), and DNA ligase IV (Dnl4-Lif1-Nej1). Much is known about the interactions that mediate the formation of each complex, but little is known about how they act together during repair. A comprehensive yeast two-hybrid screen of the NHEJ factors of S. cerevisiae revealed all known interactions within the MRX, Ku, and DNA ligase IV complexes, as well as three additional, weaker interactions between Yku80-Dnl4, Xrs2-Lif1, and Mre11-Yku80. Individual and combined deletions of the Yku80 C terminus and the Xrs2 forkhead-associated (FHA) domain were designed based on the latter two-hybrid results. These deletions synergistically blocked NHEJ but not the telomere and recombination functions of Ku and MRX, confirming that these protein regions are functionally important specifically for NHEJ. Further mutational analysis of Yku80 identified a putative C-terminal amphipathic alpha-helix that is both required for its NHEJ function and strikingly similar to a DNA-dependent protein kinase interaction motif in human Ku80. These results identify a novel role in yeast NHEJ for the poorly characterized Ku80 C-terminal and Xrs2 FHA domains, and they suggest that redundant binding of DNA ligase IV facilitates completion of this DNA repair event. PMID- 16314504 TI - Inhibited neurogenesis in JNK1-deficient embryonic stem cells. AB - The JNKs are components of stress signaling pathways but also regulate morphogenesis and differentiation. Previously, we invoked a role for the JNKs in nerve growth factor (NGF)-stimulated PC12 cell neural differentiation (L. Marek et al., J. Cell. Physiol. 201:459-469, 2004; E. Zentrich et al., J. Biol. Chem. 277:4110-4118, 2002). Herein, the role for JNKs in neural differentiation and transcriptional regulation of the marker gene, NFLC, modeled in mouse embryonic stem (ES) cells was studied. NFLC-luciferase reporters revealed the requirement for NFLC promoter sequences encompassing base pairs -128 to -98 relative to the transcriptional start site as well as a proximal cyclic AMP response element activating transcription factor binding site at -45 to -38 base pairs for transcriptional induction in NGF-treated PC12 cells and neurally differentiated ES cells. The findings reveal common promoter sequences that integrate conserved signal pathways in both PC12 cell and ES cell systems. To test the requirement for the JNK pathway in ES cell neurogenesis, ES cell lines bearing homozygous disruptions of the jnk1, jnk2, or jnk3 genes were derived and submitted to an embryoid body (EB) differentiation protocol. Neural differentiation was observed in wild-type, JNK2(-/-), and JNK3(-/-) cultures but not in JNK1(-/-) EBs. Rather, an outgrowth of cells with epithelial morphology and enhanced E-cadherin expression but low NFLC mRNA and protein was observed in JNK1(-/-) cultures. The expression of wnt-4 and wnt-6, identified inhibitors of ES cell neurogenesis, was significantly elevated in JNK1(-/-) cultures relative to wild-type, JNK2(-/-), and JNK3(-/-) cultures. Moreover, the Wnt antagonist, sFRP-2, partially rescued neural differentiation in JNK1(-/-) cultures. Thus, a genetic approach using JNK deficient ES cells reveals a novel role for JNK1 involving repression of Wnt expression in neural differentiation modeled in murine ES cells. PMID- 16314505 TI - Tyrosine phosphorylation of phosphoinositide-dependent kinase 1 by the insulin receptor is necessary for insulin metabolic signaling. AB - In L6 myoblasts, insulin receptors with deletion of the C-terminal 43 amino acids (IR(Delta43)) exhibited normal autophosphorylation and IRS-1/2 tyrosine phosphorylation. The L6 cells expressing IR(Delta43) (L6(IRDelta43)) also showed no insulin effect on glucose uptake and glycogen synthase, accompanied by a >80% decrease in insulin induction of 3-phosphoinositide-dependent protein kinase 1 (PDK-1) activity and tyrosine phosphorylation and of protein kinase B (PKB) phosphorylation at Thr(308). Insulin induced the phosphatidylinositol 3 kinase dependent coprecipitation of PDK-1 with wild-type IR (IR(WT)), but not IR(Delta43). Based on overlay blotting, PDK-1 directly bound IR(WT), but not IR(Delta43). Insulin-activated IR(WT), and not IR(Delta43), phosphorylated PDK-1 at tyrosines 9, 373, and 376. The IR C-terminal 43-amino-acid peptide (C-terminal peptide) inhibited in vitro PDK-1 tyrosine phosphorylation by the IR. Tyr-->Phe substitution prevented this inhibitory action. In the L6(hIR) cells, the C terminal peptide coprecipitated with PDK-1 in an insulin-stimulated fashion. This peptide simultaneously impaired the insulin effect on PDK-1 coprecipitation with IR(WT), on PDK-1 tyrosine phosphorylation, on PKB phosphorylation at Thr(308), and on glucose uptake. Upon insulin exposure, PDK-1 membrane persistence was significantly reduced in L6(IRDelta43) compared to control cells. In L6 cells expressing IR(WT), the C-terminal peptide also impaired insulin-dependent PDK-1 membrane persistence. Thus, PDK-1 directly binds to the insulin receptor, followed by PDK-1 activation and insulin metabolic effects. PMID- 16314506 TI - Targeting of the activation-induced cytosine deaminase is strongly influenced by the sequence and structure of the targeted DNA. AB - Activation-induced deaminase (AID) initiates immunoglobulin somatic hypermutation (SHM). Since in vitro AID was shown to deaminate cytosines on single-stranded DNA or the nontranscribed strand, it remained a puzzle how in vivo AID targets both DNA strands equally. Here we investigate the roles of transcription and DNA sequence in cytosine deamination. Strikingly different results are found with different substrates. Depending on the target sequence, the transcribed DNA strand is targeted as well as or better than the nontranscribed strand. The preferential targeting is not related to the frequency of AID hot spots. Comparison of cytosine deamination by AID and bisulfite shows different targeting patterns suggesting that AID may locally unwind the DNA. We conclude that somatic hypermutation on both DNA strands is the natural outcome of AID action on a transcribed gene; furthermore, the DNA sequence or structure and topology play major roles in targeting AID in vitro and in vivo. On the other hand, the lack of mutations in the first approximately 100 nucleotides and beyond about 1 to 2 kb from the promoter of immunoglobulin genes during SHM must be due to special conditions of transcription and chromatin in vivo. PMID- 16314507 TI - Shuttling mechanism of peroxisome targeting signal type 1 receptor Pex5: ATP independent import and ATP-dependent export. AB - Peroxisomal matrix proteins are posttranslationally imported into peroxisomes with the peroxisome-targeting signal 1 receptor, Pex5. The longer isoform of Pex5, Pex5L, also transports Pex7-PTS2 protein complexes. After unloading the cargoes, Pex5 returns to the cytosol. To address molecular mechanisms underlying Pex5 functions, we constructed a cell-free Pex5 translocation system with a postnuclear supernatant fraction from CHO cell lines. In assays using the wild type CHO-K1 cell fraction, (35)S-labeled Pex5 was specifically imported into and exported from peroxisomes with multiple rounds. (35)S-Pex5 import was also evident using peroxisomes isolated from rat liver. ATP was not required for (35)S Pex5 import but was indispensable for export. (35)S-Pex5 was imported neither to peroxisome remnants from RING peroxin-deficient cell mutants nor to those from pex14 cells lacking a Pex5-docking site. In contrast, (35)S-Pex5 was imported into the peroxisome remnants of PEX1-, PEX6-, and PEX26-defective cell mutants, including those from patients with peroxisome biogenesis disorders, from which, however, (35)S-Pex5 was not exported, thereby indicating that Pex1 and Pex6 of the AAA ATPase family and their recruiter, Pex26, were essential for Pex5 export. Moreover, we analyzed the (35)S-Pex5-associated complexes on peroxisomal membranes by blue-native polyacrylamide gel electrophoresis. (35)S-Pex5 was in two distinct, 500- and 800-kDa complexes comprising different sets of peroxins, such as Pex14 and Pex2, implying that Pex5 transited between the subcomplexes. Together, results indicated that Pex5 most likely enters peroxisomes, changes its interacting partners, and then exits using ATP energy. PMID- 16314508 TI - The essential WD40 protein Cia1 is involved in a late step of cytosolic and nuclear iron-sulfur protein assembly. AB - The assembly of cytosolic and nuclear iron-sulfur (Fe/S) proteins in yeast is dependent on the iron-sulfur cluster assembly and export machineries in mitochondria and three recently identified extramitochondrial proteins, the P loop NTPases Cfd1 and Nbp35 and the hydrogenase-like Nar1. However, the molecular mechanism of Fe/S protein assembly in the cytosol is far from being understood, and more components are anticipated to take part in this process. Here, we have identified and functionally characterized a novel WD40 repeat protein, designated Cia1, as an essential component required for Fe/S cluster assembly in vivo on cytosolic and nuclear, but not mitochondrial, Fe/S proteins. Surprisingly, Nbp35 and Nar1, themselves Fe/S proteins, could assemble their Fe/S clusters in the absence of Cia1, demonstrating that these components act before Cia1. Consequently, Cia1 is involved in a late step of Fe/S cluster incorporation into target proteins. Coimmunoprecipitation assays demonstrated a specific interaction between Cia1 and Nar1. In contrast to the mostly cytosolic Nar1, Cia1 is preferentially localized to the nucleus, suggesting an additional function of Cia1. Taken together, our results indicate that Cia1 is a new member of the cytosolic Fe/S protein assembly (CIA) machinery participating in a step after Nbp35 and Nar1. PMID- 16314509 TI - Autophosphorylation of DNA-dependent protein kinase regulates DNA end processing and may also alter double-strand break repair pathway choice. AB - Two highly conserved double-strand break (DSB) repair pathways, homologous recombination (HR) and nonhomologous end joining (NHEJ), function in all eukaryotes. How a cell chooses which pathway to utilize is an area of active research and debate. During NHEJ, the DNA-dependent protein kinase (DNA-PK) functions as a "gatekeeper" regulating DNA end access. Here, we provide evidence that DNA-PK regulates DNA end access via its own autophosphorylation. We demonstrated previously that autophosphorylation within a major cluster of sites likely mediates a conformational change that is critical for DNA end processing. Furthermore, blocking autophosphorylation at these sites inhibits a cell's ability to utilize the other major double-strand break repair pathway, HR. Here, we define a second major cluster of DNA-PK catalytic subunit autophosphorylation sites. Whereas blocking phosphorylation at the first cluster inhibits both end processing and HR, blocking phosphorylation at the second cluster enhances both. We conclude that separate DNA-PK autophosphorylation events may function reciprocally by not only regulating DNA end processing but also affecting DSB repair pathway choice. PMID- 16314510 TI - The ternary complex factor Net regulates cell migration through inhibition of PAI 1 expression. AB - Net, Elk-1, and Sap-1 are members of the ternary complex factor (TCF) subfamily of Ets transcription factors. They form ternary complexes with serum response factor (SRF) on serum response elements of immediate early genes such as c-fos and egr-1 and mediate responses to growth factors and mitogen-activated protein kinase signaling. Although the TCFs have been extensively studied as intermediates in signaling cascades, surprisingly little is known about their different target genes and physiological functions. We report that Net homozygous mutant mouse embryonic fibroblasts have a defect in cell migration. This defect results at least in part from increased expression of plasminogen activator inhibitor type 1 (PAI-1), a serine protease inhibitor (serpin) that controls extracellular proteolysis and cell matrix adhesion. The defect in cell migration can be reverted by the addition of a PAI-1 blocking antibody. Net represses PAI-1 promoter activity and binds to a specific region of the promoter containing Ets binding sites in the absence of SRF. We conclude that Net is a negative regulator of PAI-1 expression and is thereby involved in cell migration. PMID- 16314511 TI - Identification of functionally important amino acids of ribosomal protein L3 by saturation mutagenesis. AB - There is accumulating evidence that many ribosomal proteins are involved in shaping rRNA into their functionally correct conformations through RNA-protein interactions. Moreover, although rRNA seems to play the central role in all aspects of ribosome function, ribosomal proteins may be involved in facilitating communication between different functional regions in ribosome, as well as between the ribosome and cellular factors. In an effort to more fully understand how ribosomal proteins may influence ribosome function, we undertook large-scale mutational analysis of ribosomal protein L3, a core protein of the large subunit that has been implicated in numerous ribosome-associated functions in the past. A total of 98 different rpl3 alleles were genetically characterized with regard to their effects on killer virus maintenance, programmed -1 ribosomal frameshifting, resistance/hypersensitivity to the translational inhibitor anisomycin and, in specific cases, the ability to enhance translation of a reporter mRNA lacking the 5' (7)mGppp cap structure and 3' poly(A) tail. Biochemical studies reveal a correlation between an increased affinity for aminoacyl-tRNA and the extent of anisomycin resistance and a decreased peptidyltransferase activity and increased frameshifting efficiency. Immunoblot analyses reveal that the superkiller phenotype is not due to a defect in the ability of ribosomes to recruit the Ski complex, suggesting that the defect lies in a reduced ability of mutant ribosomes to distinguish between cap(+)/poly(A)(+) and cap(-)/poly(A)(-) mRNAs. The results of these analyses are discussed with regard to how protein-rRNA interactions may affect ribosome function. PMID- 16314512 TI - Forkhead box M1 regulates the transcriptional network of genes essential for mitotic progression and genes encoding the SCF (Skp2-Cks1) ubiquitin ligase. AB - The Forkhead box m1 (Foxm1) gene is critical for G(1)/S transition and essential for mitotic progression. However, the transcriptional mechanisms downstream of FoxM1 that control these cell cycle events remain to be determined. Here, we show that both early-passage Foxm1(-)(/)(-) mouse embryonic fibroblasts (MEFs) and human osteosarcoma U2OS cells depleted of FoxM1 protein by small interfering RNA fail to grow in culture due to a mitotic block and accumulate nuclear levels of cyclin-dependent kinase inhibitor (CDKI) proteins p21(Cip1) and p27(Kip1). Using quantitative chromatin immunoprecipitation and expression assays, we show that FoxM1 is essential for transcription of the mitotic regulatory genes Cdc25B, Aurora B kinase, survivin, centromere protein A (CENPA), and CENPB. We also identify the mechanism by which FoxM1 deficiency causes elevated nuclear levels of the CDKI proteins p21(Cip1) and p27(Kip1). We provide evidence that FoxM1 is essential for transcription of Skp2 and Cks1, which are specificity subunits of the Skp1-Cullin 1-F-box (SCF) ubiquitin ligase complex that targets these CDKI proteins for degradation during the G(1)/S transition. Moreover, early-passage Foxm1(-)(/)(-) MEFs display premature senescence as evidenced by high expression of the senescence-associated beta-galactosidase, p19(ARF), and p16(INK4A) proteins. Taken together, these results demonstrate that FoxM1 regulates transcription of cell cycle genes critical for progression into S-phase and mitosis. PMID- 16314513 TI - The carboxy-terminal Neh3 domain of Nrf2 is required for transcriptional activation. AB - Nrf2 is a transcription factor critical for the maintenance of cellular redox homeostasis. We have previously found that Nrf2 is a labile protein, and its activation in cells under stress involves mechanisms leading to its stabilization. As a modular protein, Nrf2 possesses distinct transactivation and DNA binding domains essential for its transcriptional activity. In this study, we found that the C-terminal "Neh3" domain of Nrf2 is also important for its activity. Deletion of the last 16 amino acids of the protein completely abolishes its ability to activate both reporter and endogenous gene expression. Using site directed mutagenesis, we have identified a stretch of amino acids within this region that are essential for its activity and that are found to be conserved across species and among other members of the CNC-bZIP family. Importantly, deletion of the final 16 amino acids of Nrf2 does not influence its dimerizing capability, DNA binding activity, or subcellular localization, although it does increase the half-life of the protein. In addition, this region was found to be important for interaction with CHD6 (a chromo-ATPase/helicase DNA binding protein) in a yeast two-hybrid screen. RNA interference-mediated knockdown of CHD6 reduced both the basal and tert-butylhydroquinone-inducible expression of NQO1, a prototypical Nrf2 target gene. These data suggest that the Neh3 domain may act as a transactivation domain and that it is possibly involved in interaction with components of the transcriptional apparatus to affect its transcriptional activity. PMID- 16314514 TI - Human TopBP1 ensures genome integrity during normal S phase. AB - Cell cycle checkpoints are essential for maintaining genomic integrity. Human topoisomerase II binding protein 1 (TopBP1) shares sequence similarity with budding yeast Dpb11, fission yeast Rad4/Cut5, and Xenopus Cut5, all of which are required for DNA replication and cell cycle checkpoints. Indeed, we have shown that human TopBP1 participates in the activation of replication checkpoint and DNA damage checkpoints, following hydroxyurea treatment and ionizing radiation. In this study, we address the physiological function of TopBP1 in S phase by using small interfering RNA. In the absence of exogenous DNA damage, TopBP1 is recruited to replicating chromatin. However, TopBP1 does not appear to be essential for DNA replication. TopBP1-deficient cells have increased H2AX phosphorylation and ATM-Chk 2 activation, suggesting the accumulation of DNA double-strand breaks in the absence of TopBP1. This leads to formation of gaps and breaks at fragile sites, 4N accumulation, and aberrant cell division. We propose that the cellular function of TopBP1 is to monitor ongoing DNA replication. By ensuring proper DNA replication, TopBP1 plays a critical role in the maintenance of genomic stability during normal S phase as well as following genotoxic stress. PMID- 16314515 TI - Antagonistic effects of Grg6 and Groucho/TLE on the transcription repression activity of brain factor 1/FoxG1 and cortical neuron differentiation. AB - Groucho (Gro)/TLE transcriptional corepressors are involved in a variety of developmental mechanisms, including neuronal differentiation. They contain a conserved C-terminal WD40 repeat domain that mediates interactions with several DNA-binding proteins. In particular, Gro/TLE1 interacts with forkhead transcription factor brain factor 1 (BF-1; also termed FoxG1). BF-1 is an essential regulator of neuronal differentiation during cerebral cortex development and represses transcription together with Gro/TLE1. Gro/TLE-related gene product 6 (Grg6) shares with Gro/TLEs a conserved WD40 repeat domain but is more distantly related at its N-terminal half. We demonstrate that Grg6 is expressed in cortical neural progenitor cells and interacts with BF-1. In contrast to Gro/TLE1, however, Grg6 does not promote, but rather suppresses, BF-1 mediated transcriptional repression. Consistent with these observations, Grg6 interferes with the binding of Gro/TLE1 to BF-1 and does not repress transcription when targeted to DNA. Moreover, coexpression of Grg6 and BF-1 in cortical progenitor cells leads to a decrease in the number of proliferating cells and increased neuronal differentiation. Conversely, Grg6 knockdown by RNA interference causes decreased neurogenesis. These results identify a new role for Grg6 in cortical neuron development and establish a functional link between Grg6 and BF-1. PMID- 16314516 TI - Amyloid precursor proteins anchor CPEB to membranes and promote polyadenylation induced translation. AB - The cytoplasmic polyadenylation element (CPE) binding factor, CPEB, is a sequence specific RNA binding protein that controls polyadenylation-induced translation in germ cells and at postsynaptic sites of neurons. A yeast two-hybrid screen with a mouse brain cDNA library identified the transmembrane amyloid precursor-like protein 1 (APLP1) as a CPEB-interacting factor. CPEB binds the small intracellular domain (ICD) of APLP1 and the related proteins APLP2 and APP. These proteins promote polyadenylation and translation by stimulating Aurora A catalyzed CPEB serine 174 phosphorylation. Surprisingly, CPEB, Maskin, CPSF, and several other factors involved in polyadenylation and translation and CPE containing RNA are all detected on membranes by cell fractionation and immunoelectron microscopy. Moreover, most of the RNA that undergoes polyadenylation does so in membrane-containing fractions. These data demonstrate a link between cytoplasmic polyadenylation and membrane association and implicate APP family member proteins as anchors for localized mRNA polyadenylation and translation. PMID- 16314517 TI - Inhibition of TFII-I-dependent cell cycle regulation by p53. AB - The multifunctional transcription factor TFII-I is tyrosine phosphorylated in response to extracellular growth signals and transcriptionally activates growth promoting genes. However, whether activation of TFII-I also directly affects the cell cycle profile is unknown. Here we show that under normal growth conditions, TFII-I is recruited to the cyclin D1 promoter and transcriptionally activates this gene. Most strikingly, upon cell cycle arrest resulting from genotoxic stress and p53 activation, TFII-I is ubiquitinated and targeted for proteasomal degradation in a p53- and ATM (ataxia telangiectasia mutated)-dependent manner. Consistent with a direct role of TFII-I in cell cycle regulation and cellular proliferation, stable and ectopic expression of wild-type TFII-I increases cyclin D1 levels, resulting in accelerated entry to and exit from S phase, and overcomes p53-mediated cell cycle arrest, despite radiation. We further show that the transcriptional regulation of cyclin D1 and cell cycle control by TFII-I are dependent on its tyrosine phosphorylation at positions 248 and 611, sites required for its growth signal-mediated transcriptional activity. Taken together, our data define TFII-I as a growth signal-dependent transcriptional activator that is critical for cell cycle control and proliferation and further reveal that genotoxic stress-induced degradation of TFII-I results in cell cycle arrest. PMID- 16314518 TI - Mammalian E4 is required for cardiac development and maintenance of the nervous system. AB - Ubiquitin conjugation typically requires three classes of enzyme: E1, E2, and E3. A fourth type of enzyme (E4), however, was recently shown to be required for the degradation of certain types of substrate in yeast. We previously identified UFD2a (also known as E4B) as an E4 in mammals. UFD2a is exclusively expressed in cardiac muscle during mouse embryonic development, but it is abundant in neurons of adult mice and is implicated in the pathogenesis of neurodegenerative disease. The precise physiological function of this enzyme has remained largely unknown, however. Here, we show that mice lacking UFD2a die in utero, manifesting marked apoptosis in the developing heart. Polyubiquitylation activity for an E4 substrate was greatly reduced in Ufd2a(-/-) mouse embryonic fibroblasts. Furthermore, Ufd2a(+/-) mice displayed axonal dystrophy in the nucleus gracilis, as well as degeneration of Purkinje cells accompanied by endoplasmic reticulum stress. These animals also developed a neurological disorder. UFD2a thus appears to be essential for the development of cardiac muscle, as well as for the protection of spinocerebellar neurons from degeneration induced by endoplasmic reticulum stress. PMID- 16314519 TI - Disruption of Sept6, a fusion partner gene of MLL, does not affect ontogeny, leukemogenesis induced by MLL-SEPT6, or phenotype induced by the loss of Sept4. AB - Septins are evolutionarily conserved GTP-binding proteins that can heteropolymerize into filaments. Recent studies have revealed that septins are involved in not only diverse normal cellular processes but also the pathogenesis of various diseases, including cancer. SEPT6 is ubiquitously expressed in tissues and one of the fusion partner genes of MLL in the 11q23 translocations implicated in acute leukemia. However, the roles of this septin in vivo remain elusive. We have developed Sept6-deficient mice that exhibited neither gross abnormalities, changes in cytokinesis, nor spontaneous malignancy. Sept6 deficiency did not cause any quantitative changes in any of the septins evaluated in this study, nor did it cause any additional changes in the Sept4-deficient mice. Even the depletion of Sept11, a close homolog of Sept6, did not affect the Sept6-null cells in vitro, thus implying a high degree of redundancy in the septin system. Furthermore, a loss of Sept6 did not alter the phenotype of myeloproliferative disease induced by MLL-SEPT6, thus suggesting that Sept6 does not function as a tumor suppressor. To our knowledge, this is the first report demonstrating that a disruption of the translocation partner gene of MLL in 11q23 translocation does not contribute to leukemogenesis by the MLL fusion gene. PMID- 16314520 TI - Phospholipase C-delta1 and -delta3 are essential in the trophoblast for placental development. AB - Phosphoinositide-specific phospholipase C (PLC) is a key enzyme in phosphoinositide turnover and is involved in a variety of physiological functions. We analyzed PLCdelta1 knockout mice and found that PLCdelta1 is required for the maintenance of skin homeostasis. However, there were no remarkable abnormalities except hair loss and runting in PLCdelta1 knockout mice, even though PLCdelta1 is broadly distributed. Here, we report that mice lacking both PLCdelta1 and PLCdelta3 died at embryonic day 11.5 (E11.5) to E13.5. PLCdelta1/PLCdelta3 double-knockout mice exhibited severe disruption of the normal labyrinth architecture in the placenta and decreased placental vascularization, as well as abnormal proliferation and apoptosis of trophoblasts in the labyrinth area. Furthermore, PLCdelta1/PLCdelta3 double-knockout embryos supplied with a normal placenta by the tetraploid aggregation method survived beyond E14.5, clearly indicating that the embryonic lethality is caused by a defect in trophoblasts. On the basis of these results, we conclude that PLCdelta1 and PLCdelta3 are essential in trophoblasts for placental development. PMID- 16314521 TI - Specific sequence features, recognized by the SMN complex, identify snRNAs and determine their fate as snRNPs. AB - The survival of motor neurons (SMN) complex is essential for the biogenesis of spliceosomal small nuclear ribonucleoproteins (snRNPs) as it binds to and delivers Sm proteins for assembly of Sm cores on the abundant small nuclear RNAs (snRNAs). Using the conserved snRNAs encoded by the lymphotropic Herpesvirus saimiri (HVS), we determined the specific sequence and structural features of RNAs for binding to the SMN complex and for Sm core assembly. We show that the minimal SMN complex-binding domain in snRNAs, except U1, is comprised of an Sm site (AUUUUUG) and an adjacent 3' stem-loop. The adenosine and the first and third uridines of the Sm site are particularly critical for binding of the SMN complex, which directly contacts the backbone phosphates of these uridines. The specific sequence of the adjacent stem (7 to 12 base pairs)-loop (4 to 17 nucleotides) is not important for SMN complex binding, but it must be located within a short distance of the 3' end of the RNA for an Sm core to assemble. Importantly, these defining characteristics are discerned by the SMN complex and not by the Sm proteins, which can bind to and assemble on an Sm site sequence alone. These findings demonstrate that the SMN complex is the identifier, as well as assembler, of the abundant class of snRNAs in cells because it is able to recognize an snRNP code that they contain. PMID- 16314522 TI - Endosomal transport of ErbB-2: mechanism for nuclear entry of the cell surface receptor. AB - The cell membrane receptor ErbB-2 migrates to the nucleus. However, the mechanism of its nuclear translocation is unclear. Here, we report a novel mechanism of its nuclear localization that involves interaction with the transport receptor importin beta1, nuclear pore protein Nup358, and a host of players in endocytic internalization. Knocking down importin beta1 using small interfering RNA oligonucleotides or inactivation of small GTPase Ran by RanQ69L, a dominant negative mutant of Ran, causes a nuclear transport defect of ErbB-2. Mutation of a putative nuclear localization signal in ErbB-2 destroys its interaction with importin beta1 and arrests nuclear translocation, while inactivation of nuclear export receptor piles up ErbB-2 within the nucleus. Additionally, blocking of internalization by a dominant-negative mutant of dynamin halts its nuclear localization. Thus, the cell membrane-embedded ErbB-2, through endocytosis using the endocytic vesicle as a vehicle, importin beta1 as a driver and Nup358 as a traffic light, migrates from the cell surface to the nucleus. This novel mechanism explains how a receptor tyrosine kinase on the cell surface can be translocated into the nucleus. This pathway may serve as a general mechanism to allow direct communication between cell surface receptors and the nucleus, and our findings thus open a new era in understanding direct trafficking between the cell membrane and nucleus. PMID- 16314523 TI - Regulation of NDR protein kinase by hydrophobic motif phosphorylation mediated by the mammalian Ste20-like kinase MST3. AB - NDR protein kinases are involved in the regulation of cell cycle progression and morphology. NDR1/NDR2 protein kinase is activated by phosphorylation on the activation loop phosphorylation site Ser281/Ser282 and the hydrophobic motif phosphorylation site Thr444/Thr442. Autophosphorylation of NDR is responsible for phosphorylation on Ser281/Ser282, whereas Thr444/Thr442 is targeted by an upstream kinase. Here we show that MST3, a mammalian Ste20-like protein kinase, is able to phosphorylate NDR protein kinase at Thr444/Thr442. In vitro, MST3 selectively phosphorylated Thr442 of NDR2, resulting in a 10-fold stimulation of NDR activity. MOB1A (Mps one binder 1A) protein further increased the activity, leading to a fully active kinase. In vivo, Thr442 phosphorylation after okadaic acid stimulation was potently inhibited by MST3KR, a kinase-dead mutant of MST3. Knockdown of MST3 using short hairpin constructs abolished Thr442 hydrophobic motif phosphorylation of NDR in HEK293F cells. We conclude that activation of NDR is a multistep process involving phosphorylation of the hydrophobic motif site Thr444/2 by MST3, autophosphorylation of Ser281/2, and binding of MOB1A. PMID- 16314524 TI - Reexamination of the role of ubiquitin-like modifier ISG15 in the phenotype of UBP43-deficient mice. AB - UBP43/USP18 was described as a specific protease that removes conjugated ubiquitin-like modifier ISG15 from target proteins. The severe phenotype of UBP43(-/-) mice characterized by premature death, brain cell injury, and deregulated STAT1 signaling was ascribed to an enhanced conjugation of ISG15. In contrast, no phenotypic changes were detected in ISG15(-/-) mice. To verify the role of ISG15 in the phenotype of UBP43(-/-) mice, we employed mice deficient for both ISG15 and UBP43. Here, we show that the phenotype of UBP43(-/-) mice was not rescued by the absence of ISG15, as evident from unchanged mortality, neurological symptoms, and occurrence of hydrocephalus. Also, the reported hypersensitivity of UBP43(-/-) mice to an interferon inducer, poly(I . C), was ISG15 independent. Furthermore, no evidence for a role of ISG15 in the modulation of STAT1 signaling or in the resistance against lymphocytic choriomeningitis virus and vesicular stomatitis virus was found. Presented results clearly demonstrate that the phenotypic alterations of UBP43(-/-) mice are not caused by the lack of ISG15 deconjugation and must be due to another, non-ISG15-mediated molecular mechanism. PMID- 16314525 TI - Deficiency in expression of the signaling protein Sin/Efs leads to T-lymphocyte activation and mucosal inflammation. AB - Our studies have concentrated on elucidating the role of the signaling protein Sin in T-lymphocyte function. We have previously shown that Sin overexpression inhibits T-lymphocyte development and activation. Here we show that Sin-deficient mice exhibit exaggerated immune responses characterized by enhanced cytokine secretion and T-cell-dependent antibody production. Excessive T-cell responses in young mice correlate with spontaneous development of inflammatory lesions in different organs of aged Sin(-/-) mice, particularly the small intestine. The intestinal inflammation is characterized by T- and B-cell infiltrates in the lamina propria, which correlate with crypt enlargement and marked villus expansion and/or damage. Similar to the human intestinal inflammatory disorder Crohn's disease (CD), and in contrast to most mouse models of mucosal inflammation, inflammatory lesions in the gastrointestinal tract of Sin(-/-) mice are restricted to the small bowel. Taken together, these results suggest that Sin regulates immune system and T-lymphocyte function and that immune system dysfunction in the absence of Sin may underlie the pathogenesis of tissue specific inflammation and enteropathies such as CD. PMID- 16314526 TI - Association of BMI1 with polycomb bodies is dynamic and requires PRC2/EZH2 and the maintenance DNA methyltransferase DNMT1. AB - Polycomb group (PcG) proteins are epigenetic chromatin modifiers involved in heritable gene repression. Two main PcG complexes have been characterized. Polycomb repressive complex 2 (PRC2) is thought to be involved in the initiation of gene silencing, whereas Polycomb repressive complex 1 (PRC1) is implicated in the stable maintenance of gene repression. Here, we investigate the kinetic properties of the binding of one of the PRC1 core components, BMI1, with PcG bodies. PcG bodies are unique nuclear structures located on regions of pericentric heterochromatin, found to be the site of accumulation of PcG complexes in different cell lines. We report the presence of at least two kinetically different pools of BMI1, a highly dynamic and a less dynamic fraction, which may reflect BMI1 pools with different binding capacities to these stable heterochromatin domains. Interestingly, PRC2 members EED and EZH2 appear to be essential for BMI1 recruitment to the PcG bodies. Furthermore, we demonstrate that the maintenance DNA methyltransferase DNMT1 is necessary for proper PcG body assembly independent of DNMT-associated histone deacetylase activity. Together, these results provide new insights in the mechanism for regulation of chromatin silencing by PcG proteins and suggest a highly regulated recruitment of PRC1 to chromatin. PMID- 16314527 TI - The Caenorhabditis elegans heterochronic regulator LIN-14 is a novel transcription factor that controls the developmental timing of transcription from the insulin/insulin-like growth factor gene ins-33 by direct DNA binding. AB - A temporal gradient of the novel nuclear protein LIN-14 specifies the timing and sequence of stage-specific developmental events in Caenorhabditis elegans. The profound effects of lin-14 mutations on worm development suggest that LIN-14 directly or indirectly regulates stage-specific gene expression. We show that LIN 14 can associate with chromatin in vivo and has in vitro DNA binding activity. A bacterially expressed C-terminal domain of LIN-14 was used to select DNA sequences that contain a putative consensus binding site from a pool of randomized double-stranded oligonucleotides. To identify candidates for genes directly regulated by lin-14, we employed DNA microarray hybridization to compare the mRNA abundance of C. elegans genes in wild-type animals to that in mutants with reduced or elevated lin-14 activity. Five of the candidate LIN-14 target genes identified by microarrays, including the insulin/insulin-like growth factor family gene ins-33, contain putative LIN-14 consensus sites in their upstream DNA sequences. Genetic analysis indicates that the developmental regulation of ins-33 mRNA involves the stage-specific repression of ins-33 transcription by LIN-14 via sequence-specific DNA binding. These results reinforce the conclusion that lin-14 encodes a novel class of transcription factor. PMID- 16314528 TI - Alterations of DNA and chromatin structures at telomeres and genetic instability in mouse cells defective in DNA polymerase alpha. AB - Telomere length is controlled by a homeostatic mechanism that involves telomerase, telomere-associated proteins, and conventional replication machinery. Specifically, the coordinated actions of the lagging strand synthesis and telomerase have been argued. Although DNA polymerase alpha, an enzyme important for the lagging strand synthesis, has been indicated to function in telomere metabolism in yeasts and ciliates, it has not been characterized in higher eukaryotes. Here, we investigated the impact of compromised polymerase alpha activity on telomeres, using tsFT20 mouse mutant cells harboring a temperature sensitive polymerase alpha mutant allele. When polymerase alpha was temperature inducibly inactivated, we observed sequential events that included an initial extension of the G-tail followed by a marked increase in the overall telomere length occurring in telomerase-independent and -dependent manners, respectively. These alterations of telomeric DNA were accompanied by alterations of telomeric chromatin structures as revealed by quantitative chromatin immunoprecipitation and immunofluorescence analyses of TRF1 and POT1. Unexpectedly, polymerase alpha inhibition resulted in a significantly high incidence of Robertsonian chromosome fusions without noticeable increases in other types of chromosomal aberrations. These results indicate that although DNA polymerase alpha is essential for genome wide DNA replication, hypomorphic activity leads to a rather specific spectrum of chromosomal abnormality. PMID- 16314529 TI - Modulation of muscle regeneration, myogenesis, and adipogenesis by the Rho family guanine nucleotide exchange factor GEFT. AB - Rho family guanine nucleotide exchange factors (GEFs) regulate diverse cellular processes including cytoskeletal reorganization, cell adhesion, and differentiation via activation of the Rho GTPases. However, no studies have yet implicated Rho-GEFs as molecular regulators of the mesenchymal cell fate decisions which occur during development and repair of tissue damage. In this study, we demonstrate that the steady-state protein level of the Rho-specific GEF GEFT is modulated during skeletal muscle regeneration and that gene transfer of GEFT into cardiotoxin-injured mouse tibialis anterior muscle exerts a powerful promotion of skeletal muscle regeneration in vivo. In order to molecularly characterize this regenerative effect, we extrapolate the mechanism of action by examining the consequence of GEFT expression in multipotent cell lines capable of differentiating into a number of cell types, including muscle and adipocyte lineages. Our data demonstrate that endogenous GEFT is transcriptionally upregulated during myogenic differentiation and downregulated during adipogenic differentiation. Exogenous expression of GEFT promotes myogenesis of C2C12 cells via activation of RhoA, Rac1, and Cdc42 and their downstream effector proteins, while a dominant-negative mutant of GEFT inhibits this process. Moreover, we show that GEFT inhibits insulin-induced adipogenesis in 3T3L1 preadipocytes. In summary, we provide the first evidence that the Rho family signaling pathways act as potential regulators of skeletal muscle regeneration and provide the first reported molecular mechanism illustrating how a mammalian Rho family GEF controls this process by modulating mesenchymal cell fate decisions. PMID- 16314530 TI - Evidence for high-capacity bidirectional glucose transport across the endoplasmic reticulum membrane by genetically encoded fluorescence resonance energy transfer nanosensors. AB - Glucose release from hepatocytes is important for maintenance of blood glucose levels. Glucose-6-phosphate phosphatase, catalyzing the final metabolic step of gluconeogenesis, faces the endoplasmic reticulum (ER) lumen. Thus, glucose produced in the ER has to be either exported from the ER into the cytosol before release into circulation or exported directly by a vesicular pathway. To measure ER transport of glucose, fluorescence resonance energy transfer-based nanosensors were targeted to the cytosol or the ER lumen of HepG2 cells. During perfusion with 5 mM glucose, cytosolic levels were maintained at approximately 80% of the external supply, indicating that plasma membrane transport exceeded the rate of glucose phosphorylation. Glucose levels and kinetics inside the ER were indistinguishable from cytosolic levels, suggesting rapid bidirectional glucose transport across the ER membrane. A dynamic model incorporating rapid bidirectional ER transport yields a very good fit with the observed kinetics. Plasma membrane and ER membrane glucose transport differed regarding sensitivity to cytochalasin B and showed different relative kinetics for galactose uptake and release, suggesting catalysis by distinct activities at the two membranes. The presence of a high-capacity glucose transport system on the ER membrane is consistent with the hypothesis that glucose export from hepatocytes occurs via the cytosol by a yet-to-be-identified set of proteins. PMID- 16314531 TI - Essential role for sphingosine kinases in neural and vascular development. AB - Sphingosine-1-phosphate (S1P), an important sphingolipid metabolite, regulates diverse cellular processes, including cell survival, growth, and differentiation. Here we show that S1P signaling is critical for neural and vascular development. Sphingosine kinase-null mice exhibited a deficiency of S1P which severely disturbed neurogenesis, including neural tube closure, and angiogenesis and caused embryonic lethality. A dramatic increase in apoptosis and a decrease in mitosis were seen in the developing nervous system. S1P(1) receptor-null mice also showed severe defects in neurogenesis, indicating that the mechanism by which S1P promotes neurogenesis is, in part, signaling from the S1P(1) receptor. Thus, S1P joins a growing list of signaling molecules, such as vascular endothelial growth factor, which regulate the functionally intertwined pathways of angiogenesis and neurogenesis. Our findings also suggest that exploitation of this potent neuronal survival pathway could lead to the development of novel therapeutic approaches for neurological diseases. PMID- 16314532 TI - The class II phosphoinositide 3-kinase C2beta is not essential for epidermal differentiation. AB - Phosphoinositide 3-kinases (PI3Ks) regulate an array of cellular processes and are comprised of three classes. Class I PI3Ks include the well-studied agonist sensitive p110 isoforms; however, the functions of class II and III PI3Ks are less well characterized. Of the three class II PI3Ks, C2alpha and C2beta are widely expressed in many tissues, including the epidermis, while C2gamma is confined to the liver. In contrast to the class I PI3K p110alpha, which is expressed throughout the epidermis, C2beta was found to be localized in suprabasal cells, suggesting a potential role for C2beta in epidermal differentiation. Overexpressing C2beta in epidermal cells in vitro induced differentiation markers. To study a role for C2beta in tissue, we generated transgenic mice overexpressing C2beta in both suprabasal and basal epidermal layers. These mice lacked epidermal abnormalities. Mice deficient in C2beta were then generated by targeted gene deletion. C2beta knockout mice were viable and fertile and displayed normal epidermal growth, differentiation, barrier function, and wound healing. To exclude compensation by C2alpha, RNA interference was then used to knock down both C2alpha and C2beta in epidermal cells simultaneously. Induction of differentiation markers was unaffected in the absence of C2alpha and C2beta. These findings indicate that class II PI3Ks are not essential for epidermal differentiation. PMID- 16314533 TI - periostin null mice exhibit dwarfism, incisor enamel defects, and an early-onset periodontal disease-like phenotype. AB - Periostin was originally identified as an osteoblast-specific factor and is highly expressed in the embryonic periosteum, cardiac valves, placenta, and periodontal ligament as well as in many adult cancerous tissues. To investigate its role during development, we generated mice that lack the periostin gene and replaced the translation start site and first exon with a lacZ reporter gene. Surprisingly, although periostin is widely expressed in many developing organs, periostin-deficient (peri(lacZ)) embryos are grossly normal. Postnatally, however, approximately 14% of the nulls die before weaning and all of the remaining peri(lacZ) nulls are severely growth retarded. Skeletal analysis revealed that trabecular bone in adult homozygous skeletons was sparse, but overall bone growth was unaffected. Furthermore, by 3 months, the nulls develop an early-onset periodontal disease-like phenotype. Unexpectedly, these mice also show a severe incisor enamel defect, although there is no apparent change in ameloblast differentiation. Significantly, placing the peri(lacZ) nulls on a soft diet that alleviated mechanical strain on the periodontal ligament resulted in a partial rescue of both the enamel and periodontal disease-like phenotypes. Combined, these data suggest that a healthy periodontal ligament is required for normal amelogenesis and that periostin is critically required for maintenance of the integrity of the periodontal ligament in response to mechanical stresses. PMID- 16314534 TI - Targeted deletion of integrin-linked kinase reveals a role in T-cell chemotaxis and survival. AB - Integrin-linked kinase (ILK) is a serine/threonine kinase that is important in cell-matrix interactions and cell signaling. To examine the role of ILK in leukocyte trafficking and survival, we generated T cell-specific ILK knockouts by breeding ILK(flox/flox) mice to transgenic mice expressing Cre recombinase under control of the Lck proximal promoter. Thymic T cells from Lck Cre(+)/ILK(flox/flox) mice had a marked reduction (>95%) in ILK protein levels. Thymic cellularity was comparable in 3- to 4-week-old mice, but a threefold diminution of thymic T cells became evident by 6 to 8 weeks of age in the T cell specific ILK knockout mice due to increased cell death of double-positive (DP) T cells. Analysis of peripheral T cells by quantitative PCR and by breeding Lck Cre(+)/ILK(flox/flox) mice to a YFP-transgenic reporter strain demonstrated an approximate 20-fold enrichment of ILK-competent cells, suggesting these cells have a competitive advantage in trafficking to and/or survival in peripheral lymphatic organs. We explored mechanisms related to altered cell trafficking and survival that might explain the decreases in thymic cellularity and enrichment for ILK-competent cells in the spleen and lymph nodes. We observed a >50% reduction in chemotaxis of ILK-deficient T cells to the chemokines CXCL12 (stromal cell-derived factor [SDF]-1alpha) and CCL19 (macrophage inflammatory protein [MIP]-3beta), as well as enhanced apoptosis of ILK-deficient cells upon stress. Signaling studies in ILK-deficient T cells demonstrated diminished phosphorylation of Akt on the activating phosphorylation site, Ser 473, and a concordant decrease in Akt kinase activity following stimulation with the chemokine SDF-1. Rac1 activation was also markedly diminished in ILK-deficient T cells following chemokine stimulation. These data extend the role of ILK to immune-cell trafficking and survival via modulation of Akt- and Rac-dependent substrates, and have implications for cell recruitment in both homeostatic and pathological processes. PMID- 16314535 TI - Human SWI/SNF generates abundant, structurally altered dinucleosomes on polynucleosomal templates. AB - Human SWI/SNF (hSWI/SNF) is an evolutionarily conserved ATP-dependent chromatin remodeling complex required for transcriptional regulation and cell cycle control. The regulatory functions of hSWI/SNF are correlated with its ability to create a stable, altered form of chromatin that constrains fewer negative supercoils than normal. Our current studies indicate that this change in supercoiling is due to the conversion of up to one-half of the nucleosomes on polynucleosomal arrays into asymmetric structures, termed "altosomes," each composed of two histone octamers and bearing an asymmetrically located region of nuclease-accessible DNA. Altosomes can be formed on chromatin containing the abundant mammalian linker histone H1 and have a unique micrococcal nuclease digestion footprint that allows their position and abundance on any DNA sequence to be measured. Over time, altosomes spontaneously revert to structurally normal but improperly positioned nucleosomes, suggesting a novel mechanism for transcriptional attenuation as well as transcriptional memory following hSWI/SNF action. PMID- 16314536 TI - Activator Gcn4p and Cyc8p/Tup1p are interdependent for promoter occupancy at ARG1 in vivo. AB - The Cyc8p/Tup1p complex mediates repression of diverse genes in Saccharomyces cerevisiae and is recruited by DNA binding proteins specific for the different sets of repressed genes. By screening the yeast deletion library, we identified Cyc8p as a coactivator for Gcn4p, a transcriptional activator of amino acid biosynthetic genes. Deletion of CYC8 confers sensitivity to an inhibitor of isoleucine/valine biosynthesis and impairs activation of Gcn4p-dependent reporters and authentic amino acid biosynthetic target genes. Deletion of TUP1 produces similar but less severe activation defects in vivo. Although expression of Gcn4p is unaffected by deletion of CYC8, chromatin immunoprecipitation assays reveal a strong defect in binding of Gcn4p at the target genes ARG1 and ARG4 in cyc8Delta cells and to a lesser extent in tup1Delta cells. The defects in Gcn4p binding and transcriptional activation in cyc8Delta cells cannot be overcome by Gcn4p overexpression but are partially suppressed in tup1Delta cells. The impairment of Gcn4p binding in cyc8Delta and tup1Delta cells is severe enough to reduce recruitment of SAGA, Srb mediator, TATA binding protein, and RNA polymerase II to the ARG1 and ARG4 promoters, accounting for impaired transcriptional activation of these genes in both mutants. Cyc8p and Tup1p are recruited to the ARG1 and ARG4 promoters, consistent with a direct role for this complex in stimulating Gcn4p occupancy of the upstream activation sequence (UAS). Interestingly, Gcn4p also stimulates binding of Cyc8p/Tup1p at the 3' ends of these genes, raising the possibility that Cyc8p/Tup1p influences transcription elongation. Our findings reveal a novel coactivator function for Cyc8p/Tup1p at the level of activator binding and suggest that Gcn4p may enhance its own binding to the UAS by recruiting Cyc8p/Tup1p. PMID- 16314537 TI - Rasgrf1 imprinting is regulated by a CTCF-dependent methylation-sensitive enhancer blocker. AB - Imprinted methylation of the paternal Rasgrf1 allele in mice occurs at a differentially methylated domain (DMD) 30 kbp 5' of the promoter. A repeated sequence 3' of the DMD regulates imprinted methylation, which is required for imprinted expression. Here we identify the mechanism by which methylation controls imprinting. The DMD is an enhancer blocker that binds CTCF in a methylation-sensitive manner. CTCF bound to the unmethylated maternal allele silences expression. CTCF binding to the paternal allele is prevented by repeat mediated methylation, allowing expression. Optimal in vitro enhancer-blocking activity requires CTCF binding sites. The enhancer blocker can be bypassed in vivo and imprinting abolished by placing an extra enhancer proximal to the promoter. Together, the repeats and the DMD constitute a binary switch that regulates Rasgrf1 imprinting. PMID- 16314538 TI - Subclinical hypothyroidism and cardiovascular disease. PMID- 16314539 TI - When is a new prediction marker useful? A consideration of lipoprotein-associated phospholipase A2 and C-reactive protein for stroke risk. PMID- 16314540 TI - Doctoring through Katrina: dedicated to all the people who suffered during a natural disaster. PMID- 16314541 TI - Subclinical hypothyroidism and the risk of heart failure, other cardiovascular events, and death. AB - BACKGROUND: Subclinical hypothyroidism has been associated with systolic and diastolic cardiac dysfunction and an elevated cholesterol level, but data on cardiovascular outcomes and death are limited. METHODS: We studied 2730 men and women, aged 70 to 79 years, with baseline thyrotropin (TSH) measurements and 4 year follow-up data to determine whether subclinical hypothyroidism was associated with congestive heart failure (CHF), coronary heart disease, stroke, peripheral arterial disease, and cardiovascular-related and total mortality. After the exclusion of participants with abnormal thyroxine levels, subclinical hypothyroidism was defined as a TSH level of 4.5 mIU/L or greater, and was further classified according to TSH levels (4.5-6.9, 7.0-9.9, and > or = 10.0 mIU/L). RESULTS: Subclinical hypothyroidism was present in 338 (12.4%) of the participants. Compared with euthyroid participants, CHF events occurred more frequently among those with a TSH level of 7.0 mIU/L or greater (35.0 vs 16.5 per 1000 person-years; P = .006), but not among those with TSH levels between 4.5 and 6.9 mIU/L. In multivariate analyses, the risk of CHF was higher among those with high TSH levels (TSH of 7.0-9.9 mIU/L: hazard ratio, 2.58 [95% confidence interval, 1.19-5.60]; and TSH of > or = 10.0 mIU/L: hazard ratio, 3.26 [95% confidence interval, 1.37-7.77]). Among the 2555 participants without CHF at baseline, the hazard ratio for incident CHF events was 2.33 (95% confidence interval, 1.10-4.96; P = .03) in those with a TSH of 7.0 mIU/L or greater. Subclinical hypothyroidism was not associated with increased risk for coronary heart disease, stroke, peripheral arterial disease, or cardiovascular-related or total mortality. CONCLUSIONS: Subclinical hypothyroidism is associated with an increased risk of CHF among older adults with a TSH level of 7.0 mIU/L or greater, but not with other cardiovascular events and mortality. Further investigation is warranted to assess whether subclinical hypothyroidism causes or worsens preexisting heart failure. PMID- 16314542 TI - Subclinical thyroid dysfunction as a risk factor for cardiovascular disease. AB - BACKGROUND: There have been few large epidemiological studies examining the association between thyroid dysfunction and cardiovascular disease. In particular, it is uncertain if subclinical hypothyroidism is a risk factor for cardiovascular disease. METHODS: Serum thyrotropin and free thyroxine concentrations were measured in 2108 archived serum samples from a 1981 community health survey in Busselton, Western Australia (Busselton Health Study). In a cross-sectional study, we examined the prevalence of coronary heart disease in subjects with and without subclinical thyroid dysfunction. In a longitudinal study, we examined the risk of cardiovascular mortality and coronary heart disease events (fatal and nonfatal combined) to the end of 2001 (excluding subjects who had coronary heart disease at baseline). RESULTS: In the cross sectional analysis, subjects with subclinical hypothyroidism (n = 119) had a significantly higher prevalence of coronary heart disease than euthyroid subjects (n = 1906) (age- and sex-adjusted prevalence odds ratio, 1.8; 95% confidence interval, 1.0-3.1; P = .04). In the longitudinal analysis of subjects with subclinical hypothyroidism (n = 101), there were 21 cardiovascular deaths observed compared with 9.5 expected (age- and sex-adjusted hazard ratio, 1.5; 95% confidence interval, 1.0-2.4; P = .08) and 33 coronary heart disease events observed compared with 14.7 expected (age- and sex-adjusted hazard ratio, 1.7; 95% confidence interval, 1.2-2.4; P < .01). The increased risk of coronary heart disease events remained significant after further adjustment for standard cardiovascular risk factors. Subjects with subclinical hyperthyroidism (n = 39) had no adverse outcomes. CONCLUSION: Subclinical hypothyroidism may be an independent risk factor for coronary heart disease. PMID- 16314543 TI - C-reactive protein and risk of cardiovascular disease in men and women from the Framingham Heart Study. AB - BACKGROUND: Determination of C-reactive protein (CRP) level has been suggested to improve cardiovascular disease (CVD) risk assessment. This study examines the utility of CRP levels to assess CVD risk in a community setting. METHODS: We performed a prospective observational cohort study on a community population sample. A total of 1949 men and 2497 women without CVD from the Framingham Heart Study underwent CVD risk factor assessment. Initial CVD events during 8 years of follow-up were recorded. RESULTS: There were 283 major CVD and 160 major coronary heart disease incident events. Age-, sex-, and multivariable-adjusted analyses generally used CRP level categories of less than 1, 1 to 3, and greater than 3 mg/L. In age- and sex-adjusted models, the traditional risk factors and elevated CRP levels indicated increased risk. The age- and sex-adjusted relative risk (RR) and 95% confidence interval (CI) of CRP level greater than 3 mg/L for major CVD was elevated (RR, 1.60; 95% CI, 1.19-2.14), with evidence of attenuation (RR, 1.22; 95% CI, 0.90-1.66) in multivariable models. The C statistic, a measure of the discriminatory capability of the prediction models, was 0.74 for prediction of major CVD with age and CRP level. In multivariable models that included traditional risk factors, the C statistic was 0.78, a value that was unchanged with the addition of CRP to the multivariable model. Similar relations were noted for major coronary heart disease events. CONCLUSION: Elevated CRP level provided no further prognostic information beyond traditional office risk factor assessment to predict future major CVD and major coronary heart disease in this population sample. PMID- 16314545 TI - Quality of care and outcomes of older patients with heart failure hospitalized in the United States and Canada. AB - BACKGROUND: Health care expenditure per person is significantly higher in the United States compared with Canada, but whether there are differences in quality of care of many conditions is unknown. We compared the process of care and outcomes of patients with heart failure, the most common cause of hospitalization for individuals 65 years and older in both countries. METHODS: We compared processes of care and 30-day and 1-year risk-standardized mortality rates among 28,521 US Medicare beneficiaries and 8180 similarly aged patients in Ontario, Canada, hospitalized with heart failure from 1998 to 2001. RESULTS: More US patients underwent left ventricular ejection fraction assessment during hospitalization compared with Canadian patients (61.2% vs 41.7%, P<.001). At discharge, patients in the United States were prescribed beta-blockers more frequently (28.7% vs 25.4%, P<.001) but angiotensin-converting enzyme inhibitors less frequently (54.3% vs 63.4%, P<.001). Among ideal candidates, prescription of beta-blockers (32.5% vs 29.7%, P = .08) or angiotensin-converting enzyme inhibitors (78.3% vs 77.6%, P = .68) was not significantly different between the 2 countries. The US patients had lower risk characteristics on admission and lower crude mortality rates at 30 days and 1 year. Thirty-day risk-standardized mortality was significantly lower for the US patients (8.9% vs 10.7%, P<.001), but 1-year risk-standardized mortality was no longer significantly different (32.2% vs 32.3%, P = .98). CONCLUSION: Patients with heart failure who are hospitalized in the United States had lower short-term mortality at 30 days, but 1-year mortality rates were not significantly different between the United States and Canada. PMID- 16314544 TI - Lipoprotein-associated phospholipase A2, high-sensitivity C-reactive protein, and risk for incident ischemic stroke in middle-aged men and women in the Atherosclerosis Risk in Communities (ARIC) study. AB - BACKGROUND: Measurement of inflammatory markers has been reported to identify individuals at increased risk for ischemic stroke. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a proinflammatory enzyme secreted by macrophages. We assessed Lp-PLA2 and C-reactive protein (CRP) levels along with traditional risk factors to examine their relation to ischemic stroke. METHODS: A proportional hazards model was used in a prospective case-cohort study of 12,762 apparently healthy middle-aged men and women in the Atherosclerosis Risk in Communities (ARIC) study who were observed for about 6 years. RESULTS: Mean Lp PLA2 and CRP levels adjusted for sex, race, and age were higher in the 194 stroke cases than the 766 noncases, whereas low-density lipoprotein cholesterol (LDL-C) level was not significantly different. Both Lp-PLA2 and CRP levels were associated with ischemic stroke after adjustment for age, sex, and race: hazard ratios were 2.23 for the highest vs the lowest tertile of Lp-PLA2 and 2.70 for CRP level higher than 3 vs lower than 1 mg/L. In a model that included smoking, systolic hypertension, lipid levels, and diabetes, Lp-PLA2 and CRP levels in the highest category were associated with hazard ratios of 1.91 (95% confidence interval, 1.15-3.18; P = .01) and 1.87 (95% confidence interval, 1.13-3.10; P = .02), respectively. Individuals with high levels of both CRP and Lp-PLA2 were at the highest risk after adjusting for traditional risk factors compared with individuals with low levels of both, whereas others were at intermediate risk. CONCLUSION: Levels of Lp-PLA2 and CRP may be complementary beyond traditional risk factors in identifying middle-aged individuals at increased risk for ischemic stroke. PMID- 16314546 TI - Ties that bind: multiple relationships between clinical researchers and the pharmaceutical industry. AB - BACKGROUND: It is believed that pharmaceutical industry sponsorship of clinical research leads to the development of multiple ties between clinicians and the pharmaceutical industry. To quantify this relationship we conducted a survey of medical specialists listed in the Medical Directory of Australia in 2002 and 2003. METHODS: A questionnaire was mailed that elicited information about all aspects of research relationships between clinicians and pharmaceutical companies. The odds of reporting multiple additional ties (financial and professional) with pharmaceutical companies by clinicians who had an active research relationship were compared with those who did not. All clinicians who returned a completed questionnaire about their research activities were included in the study. RESULTS: A questionnaire was mailed to 2120 medical specialists; 823 (39%) responded. Of these, 338 (41%) reported involvement in industry sponsored research in the previous year. They were more likely than others to have been offered industry-sponsored items or activities valued at more than 500 AU dollars (>382 US dollars; odds ratio [OR], 3.5; 95% confidence interval [CI], 2.6-4.7) and support for attending international conferences (OR, 5.4; 95% CI, 3.9-7.4). The strongest associations were seen for acting as a paid consultant to industry (OR, 9.0; 95% CI, 3.9-20.4) and for membership on advisory boards (OR, 6.9; 95% CI, 5.1-9.6). There was a strong relationship between research collaboration and accumulation of industry ties. For 1 additional tie the OR was 2.2 (95% CI, 1.2-3.8) and rose to 6.3 (95% CI, 3.5-11.1) with 3 ties and 41.8 (95% CI, 14.5-143.4) with 6 or more ties. CONCLUSIONS: Medical specialists who have research relationships with the pharmaceutical industry are much more likely to have multiple additional ties than those who do not have research relationships. Institutional review should discourage clinical researchers from developing multiple ties. PMID- 16314547 TI - Impact of antidepressant drug adherence on comorbid medication use and resource utilization. AB - BACKGROUND: Patients with depression are often nonadherent to therapy for depression and chronic comorbid conditions. METHODS: To determine whether improved antidepressant medication adherence is associated with an increased likelihood of chronic comorbid disease medication adherence and reduced medical costs, we conducted a retrospective study of patients initiating antidepressant drug therapy with evidence of dyslipidemia, coronary artery disease (CAD), or both; diabetes mellitus (DM); or CAD/dyslipidemia and DM identified from a claims database. Measures included antidepressant medication adherence, measured by medication possession ratio during 180 days without a 15-day gap before 90 days of therapy; comorbid medication adherence, measured by medication possession ratio during 1 year; and the association between improved antidepressant drug adherence and disease-specific and total medical costs. RESULTS: Of 8040 patients meeting the study criteria, those adherent to antidepressant medication were more likely to be adherent to comorbid therapy vs those nonadherent to antidepressant drug therapy (CAD/dyslipidemia: odds ratio [OR], 2.13; DM: OR, 1.82; and CAD/dyslipidemia/DM: OR, 1.45; P<.001 for all). Patients adherent to antidepressant drug therapy also had significantly lower disease-specific charges vs nonadherent patients (17% lower in CAD/dyslipidemia, P = .02; 8% lower in DM, P = .39; and 14% lower in CAD/dyslipidemia/DM, P = .38). These patients also incurred lower total medical charges (6.4% lower in CAD/dyslipidemia, P = .048; 11.8% lower in DM, P = .04; and 19.8% lower in CAD/dyslipidemia/DM, P = .03). CONCLUSIONS: Antidepressant drug adherence was associated with increased comorbid disease medication adherence and reduced total medical costs for CAD/dyslipidemia, DM, and CAD/dyslipidemia/DM. Future studies should investigate the relationship between increased adherence and costs beyond 1 year. PMID- 16314549 TI - Infectious disease hospitalizations among older adults in the United States from 1990 through 2002. AB - BACKGROUND: To understand conditions associated with substantial morbidity among older adults (aged > or = 65 years), we describe hospitalization rates and trends for overall infectious disease (ID) and for specific ID groups among older adults in the United States from January 1, 1990, through December 31, 2002. METHODS: The National Hospital Discharge Survey was used to generate hospitalization estimates from 1990 through 2002 for the US population of older adults. By using a comprehensive list of International Classification of Diseases, Ninth Revision, Clinical Modification codes associated with IDs, we identified and analyzed hospitalizations associated with specific ID and ID-related categories. RESULTS: There were approximately 21.4 million (SE, 636 000) ID hospitalizations among older adults from 1990 through 2002, and between 1990 through 1992 and 2000 through 2002, the ID hospitalization rate increased 13% from 449.4 to 507.9 hospitalizations per 10 000 older adults (P = .01). This increase was caused in part by the increasing relative contributions of patients aged 75 through 84 years and 85 years or older to the older adult ID hospitalization rate. Almost half of ID hospitalizations (46% [SE, 0.7%]) and ID-related hospital deaths (48% [SE, 1.6%]) among older adults were associated with lower respiratory tract infections from 2000 through 2002. CONCLUSIONS: The hospitalization rate for IDs increased slightly among the older adult US population during the 13-year study and was associated with the aging of the older adult population. The reduction of ID hospitalization rates among older adults could help attenuate the anticipated increase in the number of hospitalizations among older adults and should be a high priority given the projected population growth among older adults in the United States. PMID- 16314548 TI - Depression and medication adherence in outpatients with coronary heart disease: findings from the Heart and Soul Study. AB - BACKGROUND: Depression leads to adverse outcomes in patients with coronary heart disease (CHD). Medication nonadherence is a potential mechanism for the increased risk of CHD events associated with depression, but it is not known whether depression is associated with medication nonadherence in outpatients with stable CHD. METHODS: We examined the association between current major depression (assessed using the Diagnostic Interview Schedule) and self-reported medication adherence in a cross-sectional study of 940 outpatients with stable CHD. RESULTS: A total of 204 participants (22%) had major depression. Twenty-eight (14%) of 204 depressed participants reported not taking their medications as prescribed compared with 40 (5%) of 736 nondepressed participants (odds ratio [OR], 2.8; 95% confidence interval [CI], 1.7-4.7; P<.001). Twice as many depressed participants as nondepressed participants (18% vs 9%) reported forgetting to take their medications (OR, 2.4; 95% CI, 1.6-3.8; P<.001). Nine percent of depressed participants and 4% of nondepressed participants reported deciding to skip their medications (OR, 2.2; 95% CI, 1.2-4.2; P = .01). The relationship between depression and nonadherence persisted after adjustment for potential confounding variables, including age, ethnicity, education, social support, and measures of cardiac disease severity (OR, 2.2; 95% CI, 1.2-3.9; P = .009 for not taking medications as prescribed). CONCLUSIONS: Depression is associated with medication nonadherence in outpatients with CHD. Medication nonadherence may contribute to adverse cardiovascular outcomes in depressed patients. PMID- 16314550 TI - Allergic rhinitis, asthma, and atherosclerosis in the Bruneck and ARMY studies. AB - BACKGROUND: Several diseases characterized by chronic inflammation and immune activation have been linked to enhanced risk for atherosclerosis. The potential association between allergies and atherosclerosis, however, remains to be defined. METHODS: The association between common allergic diseases (allergic rhinitis and asthma) and 5-year development and progression of carotid atherosclerosis (Bruneck Study) and high intima-media thickness in carotid and femoral arteries (Atherosclerosis Risk Factors in Male Youngsters [ARMY] study) was investigated. The Bruneck Study is a prospective population-based survey of 826 men and women aged 40 to 70 years; the ARMY study is a cross-sectional evaluation of 141 men aged 17 or 18 years. RESULTS: Subjects with allergic disorders were at a significantly increased risk for high intima-media thickness in the ARMY study (odds ratio, 2.5; 95% confidence interval, 1.1-5.5; P=.03) and for atherosclerosis development and progression in the Bruneck Study (odds ratio, 3.8; 95% confidence interval, 1.4-10.2; P=.007). The associations remained significant after multivariate adjustment for a broad array of established and potential vascular risk factors. When IgE levels were substituted for the clinical allergy variable, findings were confirmed in the Bruneck Study (adjusted odds ratio, 1.7; 95% confidence interval, 1.1-8.0), for a 1-SD increase in IgE level (P=.02). CONCLUSIONS: This study documents enhanced atherosclerosis among subjects with common allergic diseases. Our findings fit well with the emerging concept that key components of allergies, such as leukotrienes or mast cells, are active in human atherogenesis and further extend the growing list of immune system-mediated and chronic inflammatory disorders that have been linked with enhanced risk for atherosclerosis. PMID- 16314551 TI - Behavioral insomnia therapy for fibromyalgia patients: a randomized clinical trial. AB - BACKGROUND: Insomnia is common and debilitating to fibromyalgia (FM) patients. Cognitive-behavioral therapy (CBT) is effective for many types of patients with insomnia, but has yet to be tested with FM patients. This study compared CBT with an alternate behavioral therapy and usual care for improving sleep and other FM symptoms. METHODS: This randomized clinical trial enrolled 47 FM patients with chronic insomnia complaints. The study compared CBT, sleep hygiene (SH) instructions, and usual FM care alone. Outcome measures were subjective (sleep logs) and objective (actigraphy) total sleep time, sleep efficiency, total wake time, sleep latency, wake time after sleep onset, and questionnaire measures of global insomnia symptoms, pain, mood, and quality of life. RESULTS: Forty-two patients completed baseline and continued into treatment. Sleep logs showed CBT treated patients achieved nearly a 50% reduction in their nocturnal wake time by study completion, whereas SH therapy- and usual care-treated patients achieved only 20% and 3.5% reductions on this measure, respectively. In addition, 8 (57%) of 14 CBT recipients met strict subjective sleep improvement criteria by the end of treatment compared with 2 (17%) of 12 SH therapy recipients and 0% of the usual care group. Comparable findings were noted for similar actigraphic improvement criteria. The SH therapy patients showed favorable outcomes on measures of pain and mental well-being. This finding was most notable in an SH therapy subgroup that self-elected to implement selected CBT strategies. CONCLUSIONS: Cognitive-behavioral therapy represents a promising intervention for sleep disturbance in FM patients. Larger clinical trials of this intervention with FM patients seem warranted. PMID- 16314552 TI - Antiretroviral therapy and the prevalence and incidence of diabetes. PMID- 16314553 TI - Inadvertent iodine excess causing thyrotoxic hypokalemic periodic paralysis. PMID- 16314554 TI - Differential loss to follow-up by insurance status in the Health and Retirement Study: implications for national estimates on health insurance coverage. PMID- 16314555 TI - Sun exposure may confound physical activity-prostate cancer association. PMID- 16314556 TI - Exercise and prostate cancer: running from AGE? PMID- 16314557 TI - Comments on the important drug interaction of warfarin and sulfamethoxazole. PMID- 16314558 TI - Interfering with interferons: Hepatitis C virus counters innate immunity. PMID- 16314559 TI - Removing intensity effects and identifying significant genes for Affymetrix arrays in macrophage migration inhibitory factor-suppressed neuroblastoma cells. AB - A semilinear in-slide model is introduced to remove the intensity effect in the scanning process. It is demonstrated that the intensity effect can be estimated accurately and removed effectively. This normalization step is vital for Affymetrix arrays to reveal relevant biological results when comparing gene expression in multiple arrays. The normalized expression ratios are analyzed further by a modified two-sample t test along with a sieved permutation scheme for computing P values. The improved specificity and sensitivity are demonstrated by using a study on the impact of macrophage migration inhibitory factor (MIF) reduction in neuroblastoma cells. With semilinear in-slide model analysis, expression of 166 genes was altered with a P value no greater than 0.001. Among those genes, 44 were altered >2-fold. MIF-regulated genes associated with tumor development including IL-8 and C-met, which are overexpressed in many tumors, were down-regulated in MIF-reduced cells. On the other hand, some tumor suppressor genes such as EPHB6, visinin-like protein 1 (VSNL-1), and BLU were up regulated in MIF-reduced cells. In addition, we demonstrated that down-regulation of MIF expression could result in a reduction in cell proliferation and tumor growth in vitro and in vivo. Our data not only demonstrate that targeting MIF expression is a promising therapeutic strategy in human neuroblastoma therapy but also indicate the MIF target genes for additional study. PMID- 16314560 TI - Reconstructing the pathways of a cellular system from genome-scale signals by using matrix and tensor computations. AB - We describe the use of the matrix eigenvalue decomposition (EVD) and pseudoinverse projection and a tensor higher-order EVD (HOEVD) in reconstructing the pathways that compose a cellular system from genome-scale nondirectional networks of correlations among the genes of the system. The EVD formulates a genes x genes network as a linear superposition of genes x genes decorrelated and decoupled rank-1 subnetworks, which can be associated with functionally independent pathways. The integrative pseudoinverse projection of a network computed from a "data" signal onto a designated "basis" signal approximates the network as a linear superposition of only the subnetworks that are common to both signals and simulates observation of only the pathways that are manifest in both experiments. We define a comparative HOEVD that formulates a series of networks as linear superpositions of decorrelated rank-1 subnetworks and the rank-2 couplings among these subnetworks, which can be associated with independent pathways and the transitions among them common to all networks in the series or exclusive to a subset of the networks. Boolean functions of the discretized subnetworks and couplings highlight differential, i.e., pathway-dependent, relations among genes. We illustrate the EVD, pseudoinverse projection, and HOEVD of genome-scale networks with analyses of yeast DNA microarray data. PMID- 16314561 TI - Zfp312 is required for subcortical axonal projections and dendritic morphology of deep-layer pyramidal neurons of the cerebral cortex. AB - Pyramidal neurons of the cerebral cortex display marked layer- and subtype specific differences in their axonal projections and dendritic morphologies. Here we show that transcription factor Zfp312 is selectively expressed by layer V and VI subcortical projection pyramidal neurons and their progenitor cells. Knocking down Zfp312 with small interfering RNAs dramatically reduced the number of subcortical axonal projections from deep-layer pyramidal neurons and altered their dendritic morphology. In contrast, misexpression of Zfp312 in cortically projecting pyramidal neurons of layers II and III induced the expression of Tbr1, a transcription factor enriched in deep-layer neurons, and the formation of ectopic subcortical axonal projections. Thus, our results indicate that transcription factor Zfp312 plays a critical role in layer- and neuronal subtype specific patterning of cortical axonal projections and dendritic morphologies. PMID- 16314562 TI - Altered differentiation of neural stem cells in fragile X syndrome. AB - Fragile X syndrome, a common form of inherited mental retardation, is caused by the absence of the fragile X mental retardation protein (FMRP) due to a mutation in the FMR1 gene. We investigated the differentiation of neural stem cells generated from the brains of fmr1-knockout (KO) mice and from postmortem tissue of a fragile X fetus. Mouse and human FMRP-deficient neurospheres generated more TuJ1-positive cells (3-fold and 5-fold, respectively) than the control neurospheres generated from normal mouse and human brains, and these cells showed morphological alterations with fewer and shorter neurites and a smaller cell body volume. The number of cells expressing glial fibrillary acidic protein and generated by these neurospheres was reduced because of increased apoptotic cell death. Furthermore, there was an increase in a population of cells with intense oscillatory Ca2+ responses to neurotransmitters in differentiated cells lacking FMRP. In addition, the number of cells in a cohort of bromodeoxyuridine-labeled newborn cells was increased in the subventricular zone of the telencephalon of the fmr1-KO mouse in vivo. These results demonstrate substantial alterations in the early maturation of FMRP-deficient neural stem cells in fragile X syndrome and in the fmr1-KO mice. PMID- 16314563 TI - Cytochrome c conformations resolved by the photon counting histogram: watching the alkaline transition with single-molecule sensitivity. AB - We apply the photon counting histogram (PCH) model, a fluorescence technique with single-molecule sensitivity, to study pH-induced conformational changes of cytochrome c. PCH is able to distinguish different protein conformations based on the brightness of a fluorophore sensitive to its local environment. We label cytochrome c through its single free cysteine with tetramethylrhodamine-5 maleimide (TMR), a fluorophore with specific brightnesses that we associate with specific protein conformations. Ensemble measurements demonstrate two different fluorescence responses with increasing pH: (i) a decrease in fluorescence intensity caused by the alkaline transition of cytochrome c (pH 7.0-9.5), and (ii) an increase in intensity when the protein unfolds (pH 9.5-10.8). The magnitudes of these two responses depend strongly on the molar ratio of TMR used to label cytochrome c. Using PCH we determine that this effect arises from the proportion of a nonfunctional conformation in the sample, which can be differentiated from the functional conformation. We further determine the causes of each ensemble fluorescence response: (i) during the alkaline transition, the fluorophore enters a dark state and discrete conformations are observed, and (ii) as cytochrome c unfolds, the fluorophore incrementally brightens, but discrete conformations are no longer resolved. Moreover, we also show that functional TMR cytochrome c undergoes a response of identical magnitude regardless of the proportion of nonfunctional protein in the sample. As expected for a technique with single-molecule sensitivity, we demonstrate that PCH can directly observe the most relevant conformation, unlike ensemble fluorometry. PMID- 16314564 TI - Dpp signaling and the induction of neoplastic tumors by caspase-inhibited apoptotic cells in Drosophila. AB - In Drosophila, stresses such as x-irradiation or severe heat shock can cause most epidermal cells to die by apoptosis. Yet, the remaining cells recover from such assaults and form normal adult structures, indicating that they undergo extra growth to replace the lost cells. Recent studies of cells in which the cell death pathway is blocked by expression of the caspase inhibitor P35 have raised the possibility that dying cells normally regulate this compensatory growth by serving as transient sources of mitogenic signals. Caspase-inhibited cells that initiate apoptosis do not die. Instead, they persist in an "undead" state in which they ectopically express the signaling genes decapentaplegic (dpp) and wingless (wg) and induce abnormal growth and proliferation of surrounding tissue. Here, using mutations to abolish Dpp and/or Wg signaling by such undead cells, we show that Dpp and Wg constitute opposing stimulatory and inhibitory signals that regulate this excess growth and proliferation. Strikingly, we also found that, when Wg signaling is blocked, unfettered Dpp signaling by undead cells transforms their neighbors into neoplastic tumors, provided that caspase activity is also blocked in the responding cells. This phenomenon may provide a paradigm for the formation of neoplastic tumors in mammalian tissues that are defective in executing the cell death pathway. Specifically, we suggest that stress events (exposure to chemical mutagens, viral infection, or irradiation) that initiate apoptosis in such tissues generate undead cells, and that imbalances in growth regulatory signals sent by these cells can induce the oncogenic transformation of neighboring cells. PMID- 16314565 TI - Global gene expression profiles reveal significant nuclear reprogramming by the blastocyst stage after cloning. AB - Nuclear transfer (NT) has potential applications in agriculture and biomedicine, but the technology is hindered by low efficiency. Global gene expression analysis of clones is important for the comprehensive study of nuclear reprogramming. Here, we compared global gene expression profiles of individual bovine NT blastocysts with their somatic donor cells and fertilized control embryos using cDNA microarray technology. The NT embryos' gene expression profiles were drastically different from those of their donor cells and closely resembled those of the naturally fertilized embryos. Our findings demonstrate that the NT embryos have undergone significant nuclear reprogramming by the blastocyst stage; however, problems may occur during redifferentiation for tissue genesis and organogenesis, and small reprogramming errors may be magnified downstream in development. PMID- 16314566 TI - Retrograde signaling from the brain to the retina modulates the termination of the light response in Drosophila. AB - A critical factor in visual function is the speed with which photoreceptors (PRs) return to the resting state when light intensity dims. Several elements subserve this process, many of which promote the termination of the phototransduction cascade. Although the known elements are intrinsic to PRs, we have found that prompt restoration to the resting state of the Drosophila electroretinogram can require effective communication between the retina and the underlying brain. The requirement is seen more dramatically with long than with short light pulses, distinguishing the phenomenon from gross disruption of the termination machinery. The speed of recovery is affected by mutations (in the Hdc and ort genes) that prevent PRs from transmitting visual information to the brain. It is also affected by manipulation (using either drugs like neostigmine or genetic tools to inactivate neurotransmitter release) of cholinergic signals that arise in the brain. Intracellular recordings support the hypothesis that PRs are the target of this communication. We infer that signaling from the retina to the optic lobe prompts a feedback signal to retinal PRs. Although the mechanism of this retrograde signaling remains to be discerned, the phenomenon establishes a previously unappreciated mode of control of the temporal responsiveness of a primary sensory neuron. PMID- 16314567 TI - Toward an anandamide transporter. PMID- 16314568 TI - Bioinformatic analyses implicate the collaborating meiotic crossover/chiasma proteins Zip2, Zip3, and Spo22/Zip4 in ubiquitin labeling. AB - Zip2 and Zip3 are meiosis-specific proteins that, in collaboration with several partners, act at the sites of crossover-designated, axis-associated recombinational interactions to mediate crossover/chiasma formation. Here, Spo22 (also called Zip4) is identified as a probable functional collaborator of Zip2/3. The molecular roles of Zip2, Zip3, and Spo22/Zip4 are unknown. All three proteins are part of a small evolutionary cohort comprising similar homologs in four related yeasts. Zip3 is shown to contain a RING finger whose structural features most closely match those of known ubiquitin E3s. Further, Zip3 exhibits major domainal homologies to Rad18, a known DNA-binding ubiquitin E3. Also described is an approach to the identification and mapping of repeated protein sequence motifs, Alignment Based Repeat Annotation (ABRA), that we have developed. When ABRA is applied to Zip2 and Spo22/Zip4, they emerge as a 14-blade WD40-like repeat protein and a 22-unit tetratricopeptide repeat protein, respectively. WD40 repeats of Cdc20, Cdh1, and Cdc16 and tetratricopeptide repeats of Cdc16, Cdc23, and Cdc27, all components of the anaphase-promoting complex, are also analyzed. These and other findings suggest that Zip2, Zip3, and Zip4 act together to mediate a process that involves Zip3-mediated ubiquitin labeling, potentially as a unique type of ubiquitin-conjugating complex. PMID- 16314569 TI - Implanted hair follicle stem cells form Schwann cells that support repair of severed peripheral nerves. AB - The hair follicle bulge area is an abundant, easily accessible source of actively growing, pluripotent adult stem cells. Nestin, a protein marker for neural stem cells, also is expressed in follicle stem cells and their immediate, differentiated progeny. The fluorescent protein GFP, whose expression is driven by the nestin regulatory element in transgenic mice, served to mark the follicle cell fate. The pluripotent nestin-driven GFP stem cells are positive for the stem cell marker CD34 but negative for keratinocyte marker keratin 15, suggesting their relatively undifferentiated state. These cells can differentiate into neurons, glia, keratinocytes, smooth muscle cells, and melanocytes in vitro. In vivo studies show the nestin-driven GFP hair follicle stem cells can differentiate into blood vessels and neural tissue after transplantation to the subcutis of nude mice. Equivalent hair follicle stem cells derived from transgenic mice with beta-actin-driven GFP implanted into the gap region of a severed sciatic nerve greatly enhance the rate of nerve regeneration and the restoration of nerve function. The follicle cells transdifferentiate largely into Schwann cells, which are known to support neuron regrowth. Function of the rejoined sciatic nerve was measured by contraction of the gastrocnemius muscle upon electrical stimulation. After severing the tibial nerve and subsequent transplantation of hair follicle stem cells, walking print length and intermediate toe spread significantly recovered, indicating that the transplanted mice recovered the ability to walk normally. These results suggest that hair follicle stem cells provide an important, accessible, autologous source of adult stem cells for regenerative medicine. PMID- 16314570 TI - Identification of a high-affinity binding site involved in the transport of endocannabinoids. AB - Phytocannabinoids, such as the principal bioactive component of marijuana, delta9 tetrahydrocannabinol, have been used for thousands of years for medical and recreational purposes. delta9-Tetrahydrocannabinol and endogenous cannabinoids (e.g., anandamide) initiate their agonist properties by stimulating the cannabinoid family of G protein-coupled receptors (CB1 and CB2). The biosynthesis and physiology of anandamide is well understood, but its mechanism of uptake (resulting in signal termination by fatty acid amide hydrolase) has been elusive. Mounting evidence points to the existence of a specific anandamide transport protein; however, no direct evidence for this protein has been provided. Here, we use a potent, competitive small molecule inhibitor of anandamide uptake (LY2318912, IC50 7.27 +/- 0.510 nM) to identify a high-affinity, saturable anandamide transporter binding site (LY2318912; K(d) = 7.62 +/- 1.18 nM, B(max) = 31.6 +/- 1.80 fmol/mg protein) that is distinct from fatty acid amide hydrolase. Systemic administration of the inhibitor into rodents elevates anandamide levels 5-fold in the brain and demonstrates efficacy in the formalin paw-licking model of persistent pain with no obvious adverse effects on motor function. Identification of the anandamide transporter binding site resolves a missing mechanistic link in endocannabinoid signaling, and in vivo results suggest that endocannabinoid transporter antagonists may provide a strategy for positive modulation of cannabinoid receptors. PMID- 16314571 TI - Solution structure of the Set2-Rpb1 interacting domain of human Set2 and its interaction with the hyperphosphorylated C-terminal domain of Rpb1. AB - The phosphorylation state of the C-terminal repeat domain (CTD) of the largest subunit of RNA polymerase II changes as polymerase transcribes a gene, and the distinct forms of the phospho-CTD (PCTD) recruit different nuclear factors to elongating polymerase. The Set2 histone methyltransferase from yeast was recently shown to bind the PCTD of elongating RNA polymerase II by means of a novel domain termed the Set2-Rpb1 interacting (SRI) domain. Here, we report the solution structure of the SRI domain in human Set2 (hSRI domain), which adopts a left turned three-helix bundle distinctly different from other structurally characterized PCTD-interacting domains. NMR titration experiments mapped the binding surface of the hSRI domain to helices 1 and 2, and Biacore binding studies showed that the domain binds preferably to [Ser-2 + Ser-5]-phosphorylated CTD peptides containing two or more heptad repeats. Point-mutagenesis studies identified five residues critical for PCTD binding. In view of the differential effects of these point mutations on binding to different CTD phosphopeptides, we propose a model for the hSRI domain interaction with the PCTD. PMID- 16314572 TI - Breaking the diffraction barrier in fluorescence microscopy at low light intensities by using reversibly photoswitchable proteins. AB - Fluorescence microscopy is indispensable in many areas of science, but until recently, diffraction has limited the resolution of its lens-based variant. The diffraction barrier has been broken by a saturated depletion of the marker's fluorescent state by stimulated emission, but this approach requires picosecond laser pulses of GW/cm2 intensity. Here, we demonstrate the surpassing of the diffraction barrier in fluorescence microscopy with illumination intensities that are eight orders of magnitude smaller. The subdiffraction resolution results from reversible photoswitching of a marker protein between a fluorescence-activated and a nonactivated state, whereby one of the transitions is accomplished by means of a spatial intensity distribution featuring a zero. After characterizing the switching kinetics of the used marker protein asFP595, we demonstrate the current capability of this RESOLFT (reversible saturable optical fluorescence transitions) type of concept to resolve 50-100 nm in the focal plane. The observed resolution is limited only by the photokinetics of the protein and the perfection of the zero. Our results underscore the potential to finally achieve molecular resolution in fluorescence microscopy by technical optimization. PMID- 16314573 TI - KCNQ1-dependent transport in renal and gastrointestinal epithelia. AB - Mutations in the gene encoding for the K+ channel alpha-subunit KCNQ1 have been associated with long QT syndrome and deafness. Besides heart and inner ear epithelial cells, KCNQ1 is expressed in a variety of epithelial cells including renal proximal tubule and gastrointestinal tract epithelial cells. At these sites, cellular K+ ions exit through KCNQ1 channel complexes, which may serve to recycle K+ or to maintain cell membrane potential and thus the driving force for electrogenic transepithelial transport, e.g., Na+/glucose cotransport. Employing pharmacologic inhibition and gene knockout, the present study demonstrates the importance of KCNQ1 K+ channel complexes for the maintenance of the driving force for proximal tubular and intestinal Na+ absorption, gastric acid secretion, and cAMP-induced jejunal Cl- secretion. In the kidney, KCNQ1 appears dispensable under basal conditions because of limited substrate delivery for electrogenic Na+ reabsorption to KCNQ1-expressing mid to late proximal tubule. During conditions of increased substrate load, however, luminal KCNQ1 serves to repolarize the proximal tubule and stabilize the driving force for Na+ reabsorption. In mice lacking functional KCNQ1, impaired intestinal absorption is associated with reduced serum vitamin B12 concentrations, mild macrocytic anemia, and fecal loss of Na+ and K+, the latter affecting K+ homeostasis. PMID- 16314574 TI - Loss of dispensable genes is not adaptive in yeast. AB - A substantial share of genes identified in yeast can be deleted without visible phenotypic effects. Current debate concentrates on the possible roles of seemingly dispensable genes. The costs of maintaining unnecessary functions has attracted little attention. The hypothesis of antagonistic pleiotropy postulates that adaptations to different constituents of the environment are likely to interfere with each other, and therefore loss of unnecessary functions is potentially advantageous. We tested an entire collection of nonessential yeast gene deletions in a benign and nutritionally rich environment in which the number of dispensable genes was particularly high. We applied a series of competition experiments that could detect differences in relative fitness of approximately 0.005. No beneficial deletions were found, except perhaps for the deletion of about a dozen genes that slightly improved competitive ability; however, a functional explanation of the fitness advantage is lacking. The paucity of beneficial gene deletions is striking because genetic adaptations to laboratory conditions are regularly observed in yeast. However, it accords with the finding that the gene contents of four species of Saccharomyces are nearly identical, despite up to 20 million years of independent evolution and extensive DNA sequence divergence. Such extreme conservation of functions would be improbable if there were periods of selection promoting the loss of temporarily dispensable genes. The evident cohesion of the yeast genomes may be their evolved feature or an intrinsic property of complex genetic systems. PMID- 16314575 TI - Assessing the spatiotemporal evolution of neuronal activation with single-trial event-related potentials and functional MRI. AB - The brain acts as an integrated information processing system, which methods in cognitive neuroscience have so far depicted in a fragmented fashion. Here, we propose a simple and robust way to integrate functional MRI (fMRI) with single trial event-related potentials (ERP) to provide a more complete spatiotemporal characterization of evoked responses in the human brain. The idea behind the approach is to find brain regions whose fMRI responses can be predicted by paradigm-induced amplitude modulations of simultaneously acquired single trial ERPs. The method was used to study a variant of a two-stimulus auditory target detection (odd-ball) paradigm that manipulated predictability through alternations of stimulus sequences with random or regular target-to-target intervals. In addition to electrophysiologic and hemodynamic evoked responses to auditory targets per se, single-trial modulations were expressed during the latencies of the P2 (170-ms), N2 (200-ms), and P3 (320-ms) components and predicted spatially separated fMRI activation patterns. These spatiotemporal matches, i.e., the prediction of hemodynamic activation by time-variant information from single trial ERPs, permit inferences about regional responses using fMRI with the temporal resolution provided by electrophysiology. PMID- 16314576 TI - The relationship between FRQ-protein stability and temperature compensation in the Neurospora circadian clock. AB - Temperature compensation is an important property of all biological clocks. In Neurospora crassa, negative-feedback regulation on the frequency (frq) gene's transcription by the FRQ protein plays a central role in the organism's circadian pacemaker. Earlier model calculations predicted that the stability of FRQ should determine the period length of Neurospora's circadian rhythm as well as the rhythm's temperature compensation. Here, we report experimental FRQ protein stabilities in frq mutants at 20 degrees C and 25 degrees C, and estimates of overall activation energies for mutant FRQ protein degradation. The results are consistent with earlier model predictions, i.e., temperature compensation of Neurospora's circadian rhythm is a highly regulated process where the stability of FRQ is an important factor in determining Neurospora's circadian period as well as the clock's temperature compensation. The partial loss of temperature compensation in frq7 and frq(S513I) mutants can be described by a simple negative feedback model (the Goodwin oscillator) when the experimentally obtained activation energies of FRQ degradation for theses mutants are incorporated into the model. PMID- 16314577 TI - The molecular structure and catalytic mechanism of a quorum-quenching N-acyl-L homoserine lactone hydrolase. AB - In many Gram-negative bacteria, including a number of pathogens such as Pseudomonas aeruginosa and Erwinia carotovora, virulence factor production and biofilm formation are linked to the quorum-sensing systems that use diffusible N acyl-L-homoserine lactones (AHLs) as intercellular messenger molecules. A number of organisms also contain genes coding for lactonases that hydrolyze AHLs into inactive products, thereby blocking the quorum-sensing systems. Consequently, these enzymes attract intense interest for the development of antiinfection therapies. However, the catalytic mechanism of AHL-lactonase is poorly understood and subject to controversy. We here report a 2.0-angstroms resolution structure of the AHL-lactonase from Bacillus thuringiensis and a 1.7-angstroms crystal structure of its complex with L-homoserine lactone. Despite limited sequence similarity, the enzyme shows remarkable structural similarities to glyoxalase II and RNase Z proteins, members of the metallo-beta-lactamase superfamily. We present experimental evidence that AHL-lactonase is a metalloenzyme containing two zinc ions involved in catalysis, and we propose a catalytic mechanism for bacterial metallo-AHL-lactonases. PMID- 16314578 TI - Structure of a group A streptococcal phage-encoded virulence factor reveals a catalytically active triple-stranded beta-helix. AB - Streptococcus pyogenes (group A Streptococcus) causes severe invasive infections including scarlet fever, pharyngitis (streptococcal sore throat), skin infections, necrotizing fasciitis (flesh-eating disease), septicemia, erysipelas, cellulitis, acute rheumatic fever, and toxic shock. The conversion from nonpathogenic to toxigenic strains of S. pyogenes is frequently mediated by bacteriophage infection. One of the key bacteriophage-encoded virulence factors is a putative "hyaluronidase," HylP1, a phage tail-fiber protein responsible for the digestion of the S. pyogenes hyaluronan capsule during phage infection. Here we demonstrate that HylP1 is a hyaluronate lyase. The 3D structure, at 1.8 angstroms resolution, reveals an unusual triple-stranded beta-helical structure and provides insight into the structural basis for phage tail assembly and the role of phage tail proteins in virulence. Unlike the triple-stranded beta-helix assemblies of the bacteriophage T4 injection machinery and the tailspike endosialidase of the Escherichia coli K1 bacteriophage K1F, HylP1 possesses three copies of the active center on the triple-helical fiber itself without the need for an accessory catalytic domain. The triple-stranded beta-helix is not simply a structural scaffold, as previously envisaged; it is harnessed to provide a 200 angstroms-long substrate-binding groove for the optimal reduction in hyaluronan viscosity to aid phage penetration of the capsule. PMID- 16314579 TI - Mutagenic specificity of endogenously generated abasic sites in Saccharomyces cerevisiae chromosomal DNA. AB - Abasic [apurinic/apyrimidinic (AP)] sites are common, noncoding DNA lesions. Despite extensive investigation, the mutational pattern they provoke in eukaryotic cells remains unresolved. We constructed Saccharomyces cerevisiae strains in which chromosomal AP sites were generated during normal cell growth by altered human uracil-DNA glycosylases that remove undamaged cytosines or thymines. The mutation target was the URA3 gene inserted near the ARS309 origin to allow defined replication polarity. Expression of the altered glycosylases caused a 7- to 18-fold mutator effect in AP endonuclease-deficient (deltaapn1) yeast, which depended highly on the known translesion synthesis enzymes Rev1 and DNA polymerase zeta. For the C-glycosylase, GC>CG transversions were the predominant mutations, followed by GC>AT transitions. AT>CG transversions predominated for the T-glycosylase. These results support a major role for Rev1 dependent dCMP insertion across from AP sites and a lesser role for dAMP insertion. Unexpectedly, there was also a significant proportion of dTMP insertions that suggest another mutational pathway at AP sites. Although replication polarity did not strongly influence mutagenesis at AP sites, for certain mutation types, there was a surprisingly strong difference between the transcribed and non-transcribed strands of URA3. The basis for this strand discrimination requires further exploration. PMID- 16314580 TI - Counting human somatic cell replications: methylation mirrors endometrial stem cell divisions. AB - Cell proliferation may be altered in many diseases, but it is uncertain exactly how to measure total numbers of divisions. Although it is impossible to count every division directly, potentially total numbers of stem cell divisions since birth may be inferred from numbers of somatic errors. The idea is that divisions are surreptitiously recorded by random errors that occur during replication. To test this "molecular clock" hypothesis, epigenetic errors encoded in certain methylation patterns were counted in glands from 30 uteri. Endometrial divisions can differ among women because of differences in estrogen exposures or numbers of menstrual cycles. Consistent with an association between mitotic age and methylation, there was an age-related increase in methylation with stable levels after menopause, and significantly less methylation was observed in lean or older multiparous women. Methylation patterns were diverse and more consistent with niche rather than immortal stem cell lineages. There was no evidence for decreased stem cell survival with aging. An ability to count lifetime numbers of stem cell divisions covertly recorded by random replication errors provides new opportunities to link cell proliferation with aging and cancer. PMID- 16314581 TI - Seeing is believing: structure of the catalytic domain of HIV-1 integrase in complex with human LEDGF/p75. PMID- 16314582 TI - Calcium extrusion is critical for cardiac morphogenesis and rhythm in embryonic zebrafish hearts. AB - Calcium entry into myocytes drives contraction of the embryonic heart. To prepare for the next contraction, myocytes must extrude calcium from intracellular space via the Na+/Ca2+ exchanger (NCX1) or sequester it into the sarcoplasmic reticulum, via the sarcoplasmic reticulum Ca2+-ATPase2 (SERCA2). In mammals, defective calcium extrusion correlates with increased intracellular calcium levels and may be relevant to heart failure and sarcoplasmic dysfunction in adults. We report here that mutation of the cardiac-specific NCX1 (NCX1h) gene causes embryonic lethal cardiac arrhythmia in zebrafish tremblor (tre) embryos. The tre ventricle is nearly silent, whereas the atrium manifests a variety of arrhythmias including fibrillation. Calcium extrusion defects in tre mutants correlate with severe disruptions in sarcomere assembly, whereas mutations in the L-type calcium channel that abort calcium entry do not produce this phenotype. Knockdown of SERCA2 activity by morpholino-mediated translational inhibition or pharmacological inhibition causes embryonic lethality due to defects in cardiac contractility and morphology but, in contrast to tre mutation, does not produce arrhythmia. Analysis of intracellular calcium levels indicates that homozygous tre embryos develop calcium overload, which may contribute to the degeneration of cardiac function in this mutant. Thus, the inhibition of NCX1h versus SERCA2 activity differentially affects the pathophysiology of rhythm in the developing heart and suggests that relative levels of NCX1 and SERCA2 function are essential for normal development. PMID- 16314583 TI - Mutation in sodium-calcium exchanger 1 (NCX1) causes cardiac fibrillation in zebrafish. AB - Cardiac fibrillation, a form of cardiac arrhythmia, is the most common cause of embolic stroke and death associated with heart failure. The molecular mechanisms underlying cardiac fibrillation are largely unknown. Here we report a zebrafish model for cardiac fibrillation. The hearts of zebrafish tremblor (tre) mutants exhibit chaotic movements and fail to develop synchronized contractions. Calcium imaging showed that normal calcium transients are absent in tre cardiomyocytes, and molecular cloning of the tre mutation revealed that the tre locus encodes the zebrafish cardiac-specific sodium-calcium exchanger (NCX) 1, NCX1h. Forced expression of NCX1h or other calcium-handling molecules restored synchronized heartbeats in tre mutant embryos in a dosage-dependent manner, demonstrating the critical role of calcium homeostasis in maintaining embryonic cardiac function. By creating mosaic zebrafish embryos, we showed that sporadic NCX1h-null cells were not sufficient to disrupt normal cardiac function, but clustered wild-type cardiomyocytes contract in unison in tre mutant hearts. These data signify the essential role of calcium homeostasis and NCX1h in establishing rhythmic contraction in the embryonic zebrafish heart. PMID- 16314586 TI - How does personality influence mortality in the elderly? PMID- 16314587 TI - Neuroticism, extraversion, and mortality in a defined population of older persons. AB - OBJECTIVE: The objective of this study was to test the association of the personality traits of neuroticism and extraversion with risk of death in old age. METHODS: A census was taken of a geographically defined urban community in Chicago, and those aged 65 years or older were invited to participate in an in home interview; 6158 (79% of those eligible) did so. The interview included brief measures of neuroticism and extraversion, medical history, and questions about current participation in cognitive, social, and physical activities. Vital status was subsequently monitored. The association of each trait with risk of death was examined in a series of accelerated failure-time models that controlled for age, sex, race, and education. RESULTS: During a mean of more than 6 years of observation, 2430 persons (39.5%) died. A high level of neuroticism (score = 27; 90th percentile) was associated with a 33% increase in risk of death compared with a low level of neuroticism (score = 9; 10th percentile). A high level of extraversion (score = 33; 90th percentile) was associated with a 21% decrease in risk of death compared with a low level (score = 18; 10th percentile). Adjustment for medical conditions and health-related variables did not substantially affect results, but adjusting for baseline levels of cognitive, social, and physical activity reduced the association of both traits with mortality. CONCLUSIONS: The results suggest that higher extraversion and lower neuroticism are associated with reduced risk of mortality in old age and that these associations are mediated in part by personality-related patterns of cognitive, social, and physical activity. PMID- 16314588 TI - Relation of childhood socioeconomic status and family environment to adult metabolic functioning in the CARDIA study. AB - OBJECTIVE: Low SES and a conflict-ridden, neglectful, or harsh family environment in childhood have been linked to a high rate of physical health disorders in adulthood. The objective of the present investigation was to evaluate a model of the pathways that may help to explain these links and to relate them to metabolic functioning (MF) in the Coronary Artery Risk Development In Young Adults (CARDIA) dataset. METHODS: Participants (n = 3225) in the year 15 assessment of CARDIA, age 33 to 45 years, completed measures of childhood socioeconomic status (SES), risky early family environment (RF), adult psychosocial functioning (PsyF, a latent factor measured by depression, hostility, positive and negative social contacts), and adult SES. Indicators of the latent factor MF were assessed, specifically, cholesterol, insulin, glucose, triglycerides, and waist circumference. RESULTS: The overall prevalence of metabolic syndrome was 9.7%. Structural equation modeling indicated that childhood SES and RF are associated with MF via their association with PsyF (standardized path coefficients: childhood SES to RF -0.13, RF to PsyF 0.44, PsyF to MF 0.09, all p < .05), but also directly (coefficient from childhood SES to MF -0.12, p < .05), with good overall model fit. When this model was tested separately for race-sex subgroups, it fit best for white women, fit well for African-American women and white men, but did not fit well for African-American men. CONCLUSIONS: These results indicate that childhood SES and early family environment contribute to metabolic functioning through pathways of depression, hostility, and poor quality of social contacts. PMID- 16314589 TI - Does subjective social status predict health and change in health status better than objective status? AB - OBJECTIVE: To examine, among middle-aged individuals, if subjective socioeconomic status (SES) predicts health status and change in health status over time better than objective SES. METHODS: Data are from the Whitehall II study, a prospective study of British civil servants. SES data are drawn from Phase 5 (1997-1999) of the study and health data from Phases 5 and 6 (2000-2001). Physical and mental component scores from the Short Form 36, the General Health Questionnaire, and self-rated health were used to assess health status. Multiple linear regressions were used to examine the relationship between SES and health and change in health status. RESULTS: Complete data were available on 5486 people. Results show both measures of SES to be global measures of SES. Both measures of SES were significantly associated with health outcomes and with decline in health status over time. However, when both objective and subjective measures of SES are entered simultaneously in the model to predict change in health status, it was only the latter that continues to be significantly associated with health and changes in health. CONCLUSIONS: Subjective SES is a better predictor of health status and decline in health status over time in middle-aged adults. These results are discussed in terms of three possible explanations: subjective SES is a more precise measure of social position, the results provide support for the hierarchy-health hypothesis, and the results could be an artifact of common method variance. PMID- 16314590 TI - Socioeconomic position, cognitive function, and clustering of cardiovascular risk factors in adolescence: findings from the Mater University Study of Pregnancy and its outcomes. AB - OBJECTIVES: The objectives of this study were to examine the extent of clustering of smoking, high levels of television watching, overweight, and high blood pressure among adolescents and whether this clustering varies by socioeconomic position and cognitive function. METHODS: This study was a cross-sectional analysis of 3613 (1742 females) participants of an Australian birth cohort who were examined at age 14. RESULTS: Three hundred fifty-three (9.8%) of the participants had co-occurrence of three or four risk factors. Risk factors clustered in these adolescents with a greater number of participants than would be predicted by assumptions of independence having no risk factors and three or four risk factors. The extent of clustering tended to be greater in those from lower-income families and among those with lower cognitive function. The age adjusted ratio of observed to expected co-occurrence of three or four risk factors was 2.70 (95% confidence interval [CI], 1.80-4.06) among those from low income families and 1.70 (95% CI, 1.34-2.16) among those from more affluent families. The ratio among those with low Raven's scores (nonverbal reasoning) was 2.36 (95% CI, 1.69-3.30) and among those with higher scores was 1.51 (95% CI, 1.19-1.92); similar results for the WRAT 3 score (reading ability) were 2.69 (95% CI, 1.85-3.94) and 1.68 (95% CI, 1.34-2.11). Clustering did not differ by sex. CONCLUSION: Among adolescents, coronary heart disease risk factors cluster, and there is some evidence that this clustering is greater among those from families with low income and those who have lower cognitive function. PMID- 16314591 TI - Social support and coronary heart disease: epidemiologic evidence and implications for treatment. AB - OBJECTIVE: The present paper reviews theories of social support and evidence for the role of social support in the development and progression of coronary heart disease (CHD). METHODS: Articles for the primary review of social support as a risk factor were identified with MEDLINE (1966-2004) and PsychINFO (1872-2004). Reviews of bibliographies also were used to identify relevant articles. RESULTS: In general, evidence suggests that low social support confers a risk of 1.5 to 2.0 in both healthy populations and in patients with established CHD. However, there is substantial variability in the manner in which social support is conceptualized and measured. In addition, few studies have simultaneously compared differing types of support. CONCLUSIONS: Although low levels of support are associated with increased risk for CHD events, it is not clear what types of support are most associated with clinical outcomes in healthy persons and CHD patients. The development of a consensus in the conceptualization and measurement of social support is needed to examine which types of support are most likely to be associated with adverse CHD outcomes. There also is little evidence that improving low social support reduces CHD events. PMID- 16314592 TI - Low perceived social support and post-myocardial infarction prognosis in the enhancing recovery in coronary heart disease clinical trial: the effects of treatment. AB - OBJECTIVE: In post hoc analyses, to examine in low perceived social support (LPSS) patients enrolled in the Enhancing Recovery in Coronary Heart Disease (ENRICHD) clinical trial (n = 1503), the pattern of social support following myocardial infarction (MI), the impact of psychosocial intervention on perceived support, the relationship of perceived support at the time of MI to subsequent death and recurrent MI, and the relationship of change in perceived support 6 months after MI to subsequent mortality. METHODS: Partner status (partner, no partner) and score (<12 = low support; >12 = moderate support) on the ENRICHD Social Support Instrument (ESSI) were used post hoc to define four levels of risk. The resulting 4 LPSS risk groups were compared on baseline characteristics, changes in social support, and medical outcomes to a group of concurrently enrolled acute myocardial infarction patients without depression or LPSS (MI comparison group, n = 408). Effects of treatment assignment on LPSS and death/recurrent MI were also examined. RESULTS: All 4 LPSS risk groups demonstrated improvement in perceived support, regardless of treatment assignment, with a significant treatment effect only seen in the LPSS risk group with no partner and moderate support at baseline. During an average 29-month follow-up, the combined end point of death/nonfatal MI was 10% in the MI comparison group and 23% in the ENRICHD LPSS patients; LPSS conferred a greater risk in unadjusted and adjusted models (HR = 1.74-2.39). Change in ESSI score and/or improvement in perceived social support were not found to predict subsequent mortality. CONCLUSIONS: Baseline LPSS predicted death/recurrent MI in the ENRICHD cohort, independent of treatment assignment. Intervention effects indicated a partner surrogacy role for the interventionist and the need for a moderate level of support at baseline for the intervention to be effective. PMID- 16314593 TI - Frequency of spousal interaction and 3-year progression of carotid artery intima medial thickness: the Pittsburgh Healthy Heart Project. AB - OBJECTIVE: We employed Ecological Momentary Assessment (EMA) real-time recording in the natural setting to determine whether spousal interaction frequency predicts 3-year progression of carotid artery intima medial thickening (IMT). METHODS: Participants were 250 healthy, older adults (M age = 61, 48% female) who, at baseline, underwent 6 days of ambulatory monitoring using electronic diaries to collect data on mood, activity, and posture, as well as current or recent (past 10 minutes) social interactions. Participants also underwent ultrasound imaging of the carotid arteries at baseline and 3-year follow-up. Spousal interaction frequency was computed as the sum of total interactions with only the spouse during the 6 days of monitoring. RESULTS: Spousal interaction frequency did not predict IMT change in the sample as a whole (p = .87). However, a sex by spousal interaction by marital adjustment interaction (p = .02) indicated that more frequent spousal interaction was associated with less IMT progression among men with better marital adjustment (p = .03). In contrast, frequent spousal interaction predicted greater IMT progression among women with better martial adjustment (p < .01). This effect lost significance when women's total social interactions (sum of all interactions) were included in the model. Total social interaction frequency was an independent predictor of IMT among women but not men. CONCLUSIONS: These findings extend those of previous research by suggesting that frequent spousal interactions may be associated with long-term cardiovascular health among happily married older men and demonstrate how sampling daily experience may enhance our understanding of the possible health benefits of marriage. PMID- 16314594 TI - The uncertain consultation and patient satisfaction: the impact of patients' illness perceptions and a randomized controlled trial on the training of physicians' communication skills. AB - OBJECTIVE: To identify predictors of patient satisfaction among a range of patient and practitioner variables. In particular, to focus on patients' illness perceptions and the impact of a randomized controlled trial on the training of physicians in general communication skills and how to treat patients presenting with poorly defined illness. METHODS: A randomized controlled follow-up study conducted in 28 general practices in Aarhus County, Denmark. Half of the physicians were randomized into an educational program on treatment of patients presenting with medically unexplained symptoms (somatization). One thousand seven hundred eighty-five general practice attenders presenting a new health problem completed questionnaires on illness perceptions, physical functioning, and mental distress before the consultation. After the consultation, a questionnaire including relational and communicative domains of patient satisfaction with the current consultation was completed. The physicians completed a questionnaire for each patient on diagnostics and prognostics. Predictors of patient satisfaction were determined by logistic regression. RESULTS: A large number of patient and practitioner variables predicted satisfaction in univariate logistic regression models. Results from a multivariate logistic model showed that the illness perceptions "uncertainty" (patient not knowing what is wrong) and "emotional representations" (the complaint making the patient feel worried, depressed, helpless, afraid, hopeless) predicted dissatisfaction at OR (CI) = 1.8 (1.3-2.4), p < .001 and OR (CI) = 1.5 (1-2.3), p = .03 respectively. Trained physicians were associated with dissatisfaction at OR (CI) 0.7 (0.5-1), p = .06 in the multivariate model. Furthermore, uncertain patients consulting a trained physician were less likely to be dissatisfied OR (CI) = 0.6 (0.3-1), p = .04. CONCLUSIONS: A randomized controlled trial on the training of general practitioners' communication skills improved patient satisfaction. Illness perceptions predict satisfaction. In particular, patients feeling uncertain and negatively emotionally involved in their health problem were more inclined to being dissatisfied with the consultation. PMID- 16314595 TI - Sexual abuse history: prevalence, health effects, mediators, and psychological treatment. AB - OBJECTIVE: Lifetime history of sexual abuse is estimated to range between 15% and 25% in the general female population. People who are sexually abused are at greater risk for a whole host of physical health disorders that may occur many years after the abusive incident(s). Despite the high prevalence of this trauma and its association with poor health status, abuse history often remains hidden within the context of medical care. The aims of this review are to determine which specific health disorders have been associated with sexual abuse in both women and men, to outline the types of sexual abuse associated with the worst health outcome, to discuss some possible explanations and mediators of the abuse/health relationship, to discuss when and how to talk about abuse within a clinical setting, and to present evidence for which psychological treatments have been shown to improve the mental health of patients with past sexual abuse. METHOD: To meet these objectives, we have reviewed a wide literature on the topic of sexual abuse. RESULTS: We demonstrate that abuse appears to be related to greater likelihood of headache and gastrointestinal, gynecologic, and panic related symptoms; that the poor health effects associated with abuse are also seen in men; that abuse involving penetration and multiple incidents appears to be the most harmful, and that exposure-type therapies with and without cognitive behavioral therapy hold promise for those with abuse history. CONCLUSION: We need more research examining psychological treatments that might be efficacious in treating the physical health problems associated with sexual abuse history. PMID- 16314596 TI - Trends in the prescribing of antidepressants following acute myocardial infarction, 1993-2002. AB - OBJECTIVE: There has been a substantial increase in the prescribing of antidepressants on a population basis and in particular serotonin reuptake inhibitors (SSRIs). SSRIs have lower cardiac toxicity than tricyclic antidepressants (TCAs). We examined how the prescribing of antidepressants to patients post-myocardial infarction (MI) changed in the decade 1993 to 2002, including the proportion accounted for by TCAs. METHODS: A population-based study cross-sectional time series analysis was conducted in which quarterly antidepressant prescription data were obtained for 1993 to 2002 for elderly Ontarians who had experienced an MI, as well as for age- and sex-matched controls with no history of MI. The number of patients varied per quarter, for a total of 68,870 post-MI patients and an equal number of matched controls. Covariates included age, gender, income, and number of medications dispensed in the past year. RESULTS: Post-MI patients were more likely to receive an antidepressant relative to controls, with an overall odds ratio (OR) of 1.34; 95% confidence interval (CI), 1.29-1.38. However, with adjustment for the number of medications received, post-MI patients were 20% less likely to receive an antidepressant relative to controls, adjusted OR = 0.81; 95% CI, 0.78-0.84. The proportion of antidepressants prescribed to post-MI patients accounted for by TCAs decreased, but the proportion of post-MI patients receiving a TCA remained stable at approximately 6%. CONCLUSIONS: Increases in the prescription of antidepressants, and in particular SSRIs, to post-MI patients reflect general population trends rather than any special importance attached to treating post-MI depression. The apparent greater likelihood that post-MI patients will receive an antidepressant is reversed when total number of medications is controlled, a proxy for medical utilization and comorbidity. PMID- 16314597 TI - Evidence for overlap between idiopathic environmental intolerance and somatoform disorders. AB - OBJECTIVE: Idiopathic environmental intolerance (IEI), also known as multiple chemical sensitivity, is a chronic, polysymptomatic condition that cannot be explained by an organic disease. Physical and psychological complaints are believed to be sustained by low levels of chemically unrelated substances in the environment. At present, it is unclear whether IEI is an environmental illness or a variant of somatoform disorders (SFD). This study examined whether IEI can be distinguished from SFD with respect to self-reported symptoms, trait anxiety, body-related cognitions, and symptom attributions. METHODS: We compared 54 subjects with IEI, 54 subjects with SFD but without IEI, and 44 subjects with neither IEI nor SFD on symptom scales, psychological questionnaires, and structured interviews for IEI, depression, anxiety, and SFD. RESULTS: More than half of the IEI subjects met Diagnostic and Statistical Manual of Mental Disorders, fourth edition criteria of SFD. This group shared both symptoms and psychological features of somatization with the SFD group. IEI subjects who did not fulfill criteria for a specific SFD were less impaired by their chemical sensitivity but differed nevertheless from nonsomatoform controls by significantly higher symptom scores, higher trait anxiety, a focus on autonomic sensations, and more pronounced somatic symptom attributions. These psychological features were significantly associated with the burden of somatic symptoms in both SFD and IEI. Furthermore, self-reported allergy but not total immunoglobulin E correlated with symptom burden in the total sample. CONCLUSIONS: The similarity of IEI and SFD regarding symptoms and psychological features of somatization support the hypothesis that IEI is a variant of SFD. PMID- 16314598 TI - Caregiving and gingival symptom reports: psychophysiologic mediators. AB - OBJECTIVE: We first assessed the association of caregiving with gingival symptom reports. We then assessed whether the observed relationship was mediated by psychophysiologic host factors. METHODS: Caregivers of spouses with Alzheimer's disease (n = 123) were compared with demographically similar noncaregiver spouses (n = 117). RESULTS: The percentage of caregivers (17%) who reported gingival symptoms was twice that of noncaregivers (8.5%) (p < .05), despite the fact that caregivers and noncaregivers did not differ in oral health care. The relationship between caregiving and gingival symptom reports was mediated by psychophysiologic variables. Caregivers were higher on hassles (p < .05), depressed mood (p < .05), and metabolic risk (insulin, glucose, obesity; p < .05) than were noncaregivers. Greater gingival symptom reports were also associated with greater hassles (p < .01), depressed mood (p < .001), and metabolic risk (p < .001). Measures of subcutaneous fat, inflammation, and frank diabetes were related to gingival symptom reports but not to caregiver status. CONCLUSIONS: A higher percentage of caregivers reported gingival symptoms than noncaregivers. These results have implications for research on aging, psychophysiology, and chronic stress. PMID- 16314599 TI - The relationship between posttraumatic stress disorder and self-reported health problems. AB - OBJECTIVE: There is increasing awareness that posttraumatic stress disorder (PTSD) influences physical health. The current study used the National Comorbidity Survey (NCS) data to determine if the physical health risk imparted by PTSD was an artifact of a broad range of empirically identified confounding variables. METHODS: Participants (n = 5877) were administered a modified version of the DSM-III-R PTSD module from the Diagnostic Interview Schedule (DIS). The prevalence of 14 classes of physical disorders was assessed along with six groups of variables predictive of negative health outcomes (demographics, perception of health, stress, health-related behaviors, insurance coverage, trauma/psychiatric history, and neuroticism). RESULTS: The PTSD group had a higher frequency of most disorders and scored higher on variables predictive of negative health outcomes. To determine if the health risk imparted by PTSD was an artifact of these confounding variables, a mixed direct-sequential binary logistic regression was computed. After controlling for sex, health perceptions, stress, health-related behaviors, insurance coverage, total trauma exposure, total number of psychiatric diagnoses, and neuroticism, persons with PTSD were more likely to report the presence of a disorder. CONCLUSIONS: Previous scholars have noted that PTSD could affect health outcomes. This study found that after controlling for a wide range of variables predictive of poor health, PTSD was predictive of poor health. PMID- 16314600 TI - African Americans show alterations in endogenous pain regulatory mechanisms and reduced pain tolerance to experimental pain procedures. AB - OBJECTIVES: To examine ethnic differences in pain sensitivity and relationship of pain tolerance to blood pressure and neuroendocrine factors. METHODS: Fifty-one African Americans (24 men, 27 women) and 55 people from other ethnic groups (primarily Caucasian; 26 men, 29 women) were tested twice for pain sensitivity to tourniquet ischemia, thermal heat, and cold pressor tests, once following mental stress and once following rest control. Resting and stress-induced blood pressure (BP), plasma norepinephrine (NE), and cortisol were assessed. RESULTS: In response to all three pain tests, African Americans had lower pain tolerance relative to Caucasian/Others after both rest and stress. Only the non-African American group showed the expected inverse relationship between BP and pain sensitivity. African Americans had lower cortisol concentrations at rest and stress and showed blunted NE and systolic BP responses to stress. Only in Caucasians/Others was the relationship seen between higher stress-induced BP, cortisol, and NE levels and greater pain tolerance. CONCLUSIONS: The results suggest that there are alterations in endogenous pain regulatory mechanisms involving BP, cortisol, and NE in African Americans. Such dysregulation may contribute to the greater rate of clinical pain symptoms they experience. It is hypothesized that greater chronic stress in African Americans may be a contributing factor to the alterations in pain regulation. PMID- 16314601 TI - Abnormal affective modulation of somatosensory brain processing among patients with fibromyalgia. AB - OBJECTIVE: It is well established that subjective pain perception can be modulated by negative mood states and that patients with chronic pain are characterized by high levels of depression and anxiety. Nevertheless, very little is known about the effects of negative mood induction on brain processing of somatosensory information in fibromyalgia. The objective of the present study was to examine the influence of two emotional states (pleasant and unpleasant) on brain activity of patients with fibromyalgia (FM; n = 27) and with musculoskeletal (MSK) pain resulting from identifiable somatic lesions (n = 16). METHODS: For this purpose, somatosensory-evoked potentials (SEPs) elicited by nonpainful pneumatic stimuli, delivered to the right and left hand following an oddball paradigm, were recorded when patients were viewing affective slides. RESULTS: As compared with patients with MSK pain, patients with FM displayed overall larger P50 amplitude to tactile stimuli. In addition, significantly larger P50 and smaller N80 amplitudes were found within patients with FM when they were viewing the unpleasant rather than the pleasant slides. CONCLUSION: Our data suggest an abnormal processing of nonpainful somatosensory information in FM, especially when somatic signals are arising from the body within an aversive stimulus context. These findings provide further support for the use of biopsychosocial models for understanding FM and other chronic pain states. PMID- 16314602 TI - Coping processes and hemostatic reactivity to acute stress in dementia caregivers. AB - BACKGROUND: A hypercoagulable stress response might contribute to the increased cardiovascular risk in Alzheimer's caregivers. OBJECTIVES: (1) To evaluate whether coping processes affect hemostatic reactivity to acute psychological stress and (2) whether these effects differ substantially between caregivers and noncaregivers. METHODS: Sixty elderly community-dwelling spousal caregivers of patients with Alzheimer's disease and 33 noncaregiving controls completed the revised Ways of Coping Questionnaire to assess approach/problem-solving versus avoidant coping processes. Participants were administered an acute stress test that required them to deliver a 3-minute speech challenge to the interviewer on an assigned topic. The hypercoagulability marker D-dimer was measured at three time points: baseline, immediately postspeech, and during recovery (15 minutes postspeech). RESULTS: Multivariate analysis of covariance revealed that subjects who endorsed greater levels of approach coping had decreased levels of D-dimer at all time points (p = .048). A significant three-way interaction between planful problem solving, caregiver status, and the temporal pattern of D-dimer was found (p = .004), indicating that caregivers with low levels of planful problem solving exhibited greater increases in D-dimer from baseline to speech and recovery time points relative to controls. No relationship between avoidant coping and D-dimer was found. CONCLUSIONS: These findings suggest the possibility that approach and problem-solving coping processes buffer the impact of acute psychological stressors on procoagulant activity. It remains to be seen whether interventions that increase approach/problem-solving processes might produce salutary effects among caregiving populations. PMID- 16314603 TI - Cancer attributions, distress, and health practices among gynecologic cancer survivors. AB - OBJECTIVE: Personal beliefs about one's medical condition have been related to health behaviors and psychological distress among individuals with serious illness. We examined whether beliefs about cause of cancer and prevention of recurrence were associated with health practices and distress in 134 long-term endometrial and cervical cancer survivors. METHODS: Participants completed questionnaires assessing depressive symptoms, anxiety, health behavior, and beliefs about factors that may have caused their cancer and prevented recurrence. RESULTS: Genetics/heredity was rated as the most important cancer cause, followed by stress, God's will, hormones, and environmental factors. Medical screening was rated as most important in preventing recurrence, followed by positive attitude and prayer. Stronger causal attributions were generally associated with elevated depressive symptomatology and anxiety, but women citing potentially controllable causes were more likely to be practicing healthy behaviors. Similarly, women citing health behaviors as important in preventing recurrence reported greater anxiety but were more likely to practice positive health behaviors. Health behavior and lifestyle attributions interacted with health practices in predicting distress. For example, among women who had not made positive dietary changes, rating lifestyle as important in preventing recurrence was associated with greater distress, whereas among women who had made a positive change in diet, this belief was associated with less distress. CONCLUSIONS: Results suggest that stronger attributions are associated with greater distress, but engaging in behavior believed to be important in preventing cancer or recurrence may ameliorate this distress. PMID- 16314604 TI - Randomized placebo-controlled trial of a selective serotonin reuptake inhibitor in the treatment of nondepressed tinnitus subjects. AB - OBJECTIVE: To assess the efficacy of a selective serotonin reuptake inhibitor (paroxetine) for relief of tinnitus. DESIGN: One hundred twenty tinnitus sufferers participated in a randomized double-blind placebo-controlled trial. Paroxetine or placebo was increased to a maximally tolerated dose (up to 50 mg/day), and patients were treated for a total of 31 days at the maximal dose. METHODS: Patients with chronic tinnitus were recruited from our university-based specialty clinic by referral from otolaryngologists and audiologists in the local community and by advertisement. Patients with psychotic or substance use disorders or suicidal ideation were excluded, as were those using psychoactive medications (this resulted in only 1 subject with major depression in the study) or any other medications that interact with paroxetine and those with inability to hear at one's tinnitus sensation level. Fifty-eight percent of patients were male, 92% were Caucasian, and the average age was 57. OUTCOMES MEASURES: Tinnitus matching, the Tinnitus Handicap Questionnaire, the question: How severe (bothered, aggravating) is your tinnitus? Quality of Well-Being and other psychological questionnaires. RESULTS: Paroxetine was not statistically superior to placebo on the following tinnitus measures (tinnitus matching, 5- or 10-db drop, Tinnitus Handicap Questionnaire, quality of well-being measures, how severe, how bothered, positive change). There was a significant improvement in the single item question, How aggravating is your tinnitus? for those in the paroxetine group compared with the placebo group. CONCLUSIONS: These results suggest that the majority of individuals in this study did not benefit from paroxetine in a consistent fashion. Further work remains to be done to determine if subgroups of patients (e.g., those who tolerate higher doses, those who are depressed) may benefit. PMID- 16314605 TI - Psychiatric disorders and asthma outcomes among high-risk inner-city patients. AB - OBJECTIVE: The purpose of this study was to examine the rate of psychiatric disorders among patients attending an ethnically diverse, inner-city asthma clinic for an initial visit and assess the association between psychiatric disorders and asthma morbidity. METHODS: A semistructured psychological interview was conducted to assess for psychiatric diagnoses. A pulmonary physician, who was blind to psychiatric disorder, established diagnosis of asthma based on national guidelines. RESULTS: Sixty-four percent of 85 participants received at least 1 psychiatric diagnosis. The pulmonary physician rated patients with a psychiatric disorder as achieving fewer goals (M = 2.3 +/- 1.3) for asthma control than patients without a psychiatric disorder (M = 3.6 +/- 1.5, p = .0002). Patients with a psychiatric diagnosis more frequently reported an emergency room visit for asthma during the past 6 months (OR = 4.89; 95% CI, 1.76-13.39) and greater use of short-acting beta2-agonist medication (M = 1.5 +/- 0.9 canisters per month) than patients without a psychiatric diagnosis (M = 0.9 +/- 0.8, p = .003). These findings were independent of demographics, health insurance, and asthma severity. No differences emerged between patients with and without a mental disorder on percent predicted FEV1. Patients with a psychiatric disorder reported a higher severity level for asthma symptoms than the severity level indicated by their pulmonary function in comparison to patients without a psychiatric diagnosis (OR = 3.52; 95% CI, 1.23-10.10). Health insurance appeared to be a confounding factor in this relationship. CONCLUSION: A high rate of psychiatric disorders was found among inner-city asthma patients. Psychiatric diagnoses were associated with greater perceived impairment from asthma but not objective measurement of pulmonary function. PMID- 16314606 TI - The patients' illness perceptions and the use of primary health care. AB - OBJECTIVE: To investigate if primary care patients' perceptions of a current health problem were associated with use of health care. METHOD: One thousand seven hundred eighty-five patients presenting a new health problem to 1 of 38 physicians from 28 general practices in Aarhus County, Denmark. Patients completed a questionnaire on their illness perceptions and emotional distress before the consultation. The physicians completed a questionnaire for each patient on diagnostics and prognostics. Register data on primary health care utilization 3 years before and 2 years after baseline were obtained. Odds ratios were estimated to examine associations between previous health care use and illness perceptions. Linear regression analysis was performed to examine if illness perceptions predicted later health care use. RESULTS: Previous use: Higher use was associated with psychosocial, stress, and lifestyle attributions. Accident/chance attributions were associated with higher use for patients with a chronic disorder but with lower use for patients without a chronic disorder. A strong illness identity (number of self-reported symptoms), illness worry, a long timeline perspective, a belief that the symptoms would have serious consequences, and all emotional distress variables were associated with higher use. Use during follow-up: Infection/lowered immunity attributions were associated with higher use for patients with a chronic disorder, whereas psychosocial and lifestyle attributions were associated with higher use for all patients. Illness worry and all emotional distress variables predicted higher health care use. A strong illness identity, a long timeline perspective, a belief in serious consequences, and stress and accident/chance attributions were among the strongest predictors of health care use in a multivariate model including all variables. CONCLUSIONS: Patients' perceptions of a current health problem are associated with health care use and may offer an obvious starting point for a biopsychosocial approach in primary care. PMID- 16314607 TI - Patterns of physical symptoms and relationships with psychosocial factors in adolescents. AB - OBJECTIVES: Physical symptoms are common in otherwise healthy adolescents. This study sought to identify meaningful patterns of multiple physical symptoms and to examine relationships between the patterns and psychosocial factors. One-year stability of symptom patterns and factors contributing to stability were also examined. METHODS: This secondary data analysis used longitudinal data from a nationally representative sample of adolescents in grades 7 through 12 (n = 9,141) who participated in the National Longitudinal Study of Adolescent Health during 1994 to 1996. Ten selected physical symptoms (i.e., headache, stomachaches, fatigue) were used to construct clusters. Each cluster was compared in regard to demographic factors, psychological adjustment and interpersonal relationships. RESULTS: K-means in combination with Ward method clustered the sample into 4 groups according to the overall patterns of the 10 symptoms: nonsymptom (41%), moderate symptom (38%), high symptom (19%) and extreme symptom (2%). Adolescents in higher symptom clusters were more likely to be girls, nonwhites, or from families on welfare and reported high depressive symptoms, low self-esteem, and poor perceptions of parental affection and friendship quality. About 16% in clusters with lower symptom patterns develop somatizing patterns in Wave II; new onset was predicted by gender, younger age, and depressive symptoms. CONCLUSION: Symptom patterns characterized by overall high frequencies of multiple symptoms may indicate somatization. This study also suggests that adolescents with a somatizing tendency are more likely to experience psychological and interpersonal difficulties, and girls and younger adolescents are more vulnerable. Targeted prevention programs are needed for these vulnerable individuals by addressing their psychosocial functioning. PMID- 16314608 TI - Psychosocial factors predict CD4 and viral load change in men and women with human immunodeficiency virus in the era of highly active antiretroviral treatment. AB - OBJECTIVE: Most previous longitudinal studies demonstrating relationships between psychosocial variables and human immunodeficiency virus (HIV) disease progression utilized samples of gay men accrued before the era of highly active antiretroviral treatment (HAART), without including viral load (VL) as an indicator of disease progression or assessing the impact of medication adherence. This study sought to determine whether psychosocial variables would predict both CD4 and VL changes in a diverse sample assessed entirely during the era of HAART and accounting for adherence effects. METHODS: This longitudinal study assessed a multiethnic HIV+ sample (n = 177) of men and women in the midrange of illness (CD4 number between 150 and 500; no previous acquired immunodeficiency syndrome [AIDS]-defining symptom) every 6 months for 2 years. Hierarchical linear modeling was used to model change in CD4 and VL controlling for sociodemographics (age, gender, ethnicity, education) and medical variables (baseline CD4/VL, antiretroviral medications at each time point, adherence). RESULTS: Baseline depression, hopelessness, and education predicted the slope of CD4 and VL. Avoidant coping and life event stress predicted VL change. Cumulative variables produced stronger relationships (depression, avoidant coping, and hopelessness with CD4/VL slope and life events stress with VL slope). High cumulative depression and avoidant coping were associated with approximately twice the rate of decline in CD4 as low scorers and greater relative increases in VL. Social support was not significantly related to CD4 or VL slope. CONCLUSIONS: Psychosocial factors contribute significantly to the variance in HIV disease progression (assessed through CD4 number and VL) in a diverse sample, accounting for adherence and do so in the era of HAART. PMID- 16314610 TI - Pediatric palliative care: relationships matter and so does pain control. PMID- 16314611 TI - The chemotherapy/radiation balance in advanced Hodgkin's lymphoma: overweight which side? PMID- 16314612 TI - Hormone replacement therapy and decreased lung cancer survival. PMID- 16314613 TI - New targets, therapies, and toxicities: lessons to be learned. PMID- 16314614 TI - Efaproxiral: should we hold our breath? PMID- 16314615 TI - Comparison of ABVD and alternating or hybrid multidrug regimens for the treatment of advanced Hodgkin's lymphoma: results of the United Kingdom Lymphoma Group LY09 Trial (ISRCTN97144519). AB - PURPOSE: To perform an open-label, randomized, controlled trial comparing treatment with doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) with two multidrug regimens (MDRs) for advanced Hodgkin's lymphoma (HL). PATIENTS AND METHODS: Eight hundred seven patients with advanced HL (stage III to IV, or earlier stage with systemic symptoms or bulky disease) were randomly assigned between ABVD and MDR specified before randomization as alternating chlorambucil, vinblastine, procarbazine, and prednisolone (ChlVPP) with prednisolone, doxorubicin, bleomycin, vincristine, and etoposide (PABIOE), or hybrid ChlVPP/etoposide, vincristine, and doxorubicin (EVA). Radiotherapy was planned for incomplete response or initial bulk disease. RESULTS: At 52 months median follow-up, 212 event-free survival (EFS) events (disease progression or any death) were reported. In the primary comparison, at 3 years EFS was 75% (95% CI, 71% to 79%) for ABVD and 75% (95% CI, 70% to 79%) for MDRs (hazard ratio [HR] = 1.05; 95% CI, 0.8 to 1.37; HR more than 1.0 favors ABVD). The 3-year overall survival (OS) rates were 90% (95% CI, 87% to 93%) in patients allocated ABVD and 88% (95% CI, 84% to 91%) in patients allocated MDRs (HR = 1.22; 95% CI, 0.84 to 1.77). Patients receiving MDRs experienced more grade 3/4 infection, mucositis, and neuropathy. One occurrence of myelodysplastic syndrome was reported, but no acute leukemia was reported. When the two MDRs are compared separately with ABVD, neither the alternating nor the hybrid regimen showed a statistically significant difference from ABVD for EFS or OS. Subgroup analysis suggested that MDRs may be associated with poorer outcomes in older patients (heterogeneity test of OS older or younger than 45 years, P = .020). CONCLUSION: There was no evidence of significant difference in EFS or OS between ABVD and MDRs in the trial overall or if the two MDR versus ABVD comparisons are considered separately. ABVD remains the standard for treatment of advanced HL. PMID- 16314616 TI - Hormone replacement therapy is associated with decreased survival in women with lung cancer. AB - PURPOSE: Lung cancer is the leading cause of cancer-related death in women. Hormone replacement therapy (HRT) is frequently prescribed to postmenopausal women, but there is little data on its effect on lung cancer. Hence, we conducted a retrospective study to examine the impact of HRT on the natural history of lung cancer. METHODS: We conducted a retrospective chart review of women diagnosed with lung cancer between January 1994 and December 1999. Data collected included age, stage, past history of cancer, smoking history, family history of cancer, HRT use, treatment, and overall survival. The effects of various clinical features on survival were examined using Cox proportional hazards regression models. RESULTS: Four hundred ninety-eight women (median age, 67 years; range, 31 to 93 years) with lung cancer were included. A history of smoking was present in 429 women (86%), whereas 86 women (17%) had taken HRT. Women with lung cancer who received HRT were younger than women with lung cancer who never received HRT (63 v 68 years old, respectively; P < .0001). Overall survival was significantly higher in patients with no HRT compared with patients who received HRT (79 v 39 months, respectively; hazard ratio = 1.97; 95% CI, 1.14 to 3.39). This effect seemed to be more pronounced in women with a smoking history. CONCLUSION: HRT may affect outcomes from lung cancer adversely. Further studies examining the role of HRT use on outcomes from lung cancer, especially in women with a history of smoking, are urgently needed to clarify this important problem. PMID- 16314617 TI - Safety, pharmacokinetic, and antitumor activity of SU11248, a novel oral multitarget tyrosine kinase inhibitor, in patients with cancer. AB - PURPOSE: To establish the safety, pharmacokinetics, and recommended dose of sunitinib, a novel oral multitargeting tyrosine kinase inhibitor with antiangiogenic and antitumor properties, in patients with advanced malignancies. PATIENTS AND METHODS: Sunitinib was given orally for 4 weeks every 6 weeks. RESULTS: Twenty-eight patients received doses ranging from 15 to 59 mg/m2 (ranging from 50 mg every other day to 150 mg/d). Dose-limiting toxicities reported at the maximum-tolerated doses > or = 75 mg/d were reversible grade 3 fatigue, grade 3 hypertension, and grade 2 bullous skin toxicity. Therefore, the recommended dose was 50 mg/d. At this dose, the main adverse effects were sore mouth, edema, and thrombocytopenia. Hair discoloration and yellow coloration of the skin were observed at doses > or = 50 mg/d. Pharmacokinetic data indicate that potentially active target plasma concentrations > or = 50 ng/mL can be achieved with moderate interpatient variability and a long half-life compatible with a single daily dosing. Six objective responses were observed in three renal cell carcinomas, one neuroendocrine tumor, one stromal tumor, and one unknown primary adenocarcinoma patient. At higher doses (> or = 75 mg/d), tumor responses were often associated with reduced intratumoral vascularization and central tumor necrosis, eventually resulting in organ perforation or fistula. CONCLUSION: At the dose of 50 mg/d (4 weeks on, 2 weeks off), sunitinib displays manageable toxicity. Antitumor activity supports further studies in patients with renal cell carcinoma, gastrointestinal, neuroendocrine, and stromal tumors. Future studies may consider including prospective imaging techniques such as high frequency ultrasound to monitor tumor density. PMID- 16314618 TI - Allogeneic stem-cell transplantation using a reduced-intensity conditioning regimen has the capacity to produce durable remissions and long-term disease-free survival in patients with high-risk acute myeloid leukemia and myelodysplasia. AB - PURPOSE: The toxicity of allogeneic stem-cell transplantation can be substantially reduced using a reduced-intensity conditioning (RIC) regimen. This has increased the proportion of patients with myeloid malignancies eligible for allogeneic transplantation. However, the capacity of RIC allografts to produce durable remissions in patients with acute myeloid leukemia (AML) and myelodysplasia (MDS) has not yet been defined, and consequently, the role of RIC allografts in the management of these diseases remains conjectural. PATIENTS AND METHODS: Seventy-six patients with high-risk AML or MDS received an allograft using a fludarabine/melphalan RIC regimen incorporating alemtuzumab. The median age of the cohort was 52 years (range, 18 to 71 years). RESULTS: The 100-day transplantation-related mortality rate was 9%, and no patient developed greater than grade 2 graft-versus-host disease. With a median follow-up of 36 months (range, 13 to 70 months), 27 patients were alive and in remission, with 3-year actuarial overall survival (OS) and disease-free survival (DFS) rates of 41% and 37%, respectively. The 3-year OS and DFS rates of patients with AML in complete remission at the time of transplantation were 48% and 42%, respectively. Disease relapse was the most common cause of treatment failure and occurred at a median time of 6 months after transplantation. All but one patient destined to relapse did so within 24 months of transplantation. CONCLUSION: The extended follow-up in this series identifies a high risk of early disease relapse but provides evidence that RIC allografts can produce sustained DFS in a significant number of patients with AML who would be ineligible for allogeneic transplantation with myeloablative conditioning. PMID- 16314619 TI - Phase III study of efaproxiral as an adjunct to whole-brain radiation therapy for brain metastases. AB - PURPOSE: To determine whether efaproxiral, an allosteric modifier of hemoglobin, improves survival in patients with brain metastases when used as an adjunct to whole-brain radiation therapy (WBRT). PATIENTS AND METHODS: Patients with brain metastases from solid tumors and a Karnofsky performance score of > or = 70 were randomly assigned to receive WBRT with supplemental oxygen and either efaproxiral at 75 or 100 mg/kg (efaproxiral arm) or no efaproxiral (control arm). The primary end point was survival. RESULTS: The study consisted of 515 eligible patients (efaproxiral arm, n = 265; control arm, n = 250). The median survival time (MST) was 5.4 months for the efaproxiral arm versus 4.4 months for the control arm (hazard ratio [HR] = 0.87; P = .16). For the subgroup of patients with non-small cell lung cancer (NSCLC) or breast cancer, the MST was 6.0 and 4.4 months, respectively (HR = 0.82; P = .07). Cox multiple regression analysis demonstrated a significant reduction in the risk of death for the efaproxiral arm in both primary populations. Further analysis indicated that the benefit may be restricted to the subgroup of patients with breast cancer. Response rates (radiographic complete response plus partial response) improved by 7% (P = .10) and 13% (P = .01) for all patients and for NSCLC and breast cancer patients in the efaproxiral arm, respectively. The most common severe adverse event in patients treated with efaproxiral was hypoxemia, which was reversible and effectively managed with supplemental oxygen in most patients. CONCLUSION: The addition of efaproxiral, a noncytotoxic radiation sensitizer, to WBRT may improve response rates and survival in patients with brain metastases, particularly metastases from breast cancer. A confirmatory trial for breast cancer patients has been initiated. PMID- 16314620 TI - Osteonecrosis of the jaw in cancer after treatment with bisphosphonates: incidence and risk factors. AB - PURPOSE: Osteonecrosis of the jaw (ONJ) has been associated recently with the use of pamidronate and zoledronic acid. We studied the incidence, characteristics, and risk factors for the development of ONJ among patients treated with bisphosphonates for bone metastases. PATIENTS AND METHODS: ONJ was assessed prospectively since July 2003. The first bisphosphonate treatment among patients with ONJ was administered in 1997. Two hundred fifty-two patients who received bisphosphonates since January 1997 were included in this analysis. RESULTS: Seventeen patients (6.7%) developed ONJ: 11 of 111 (9.9%) with multiple myeloma, two of 70 (2.9%) with breast cancer, three of 46 (6.5%) with prostate cancer, and one of 25 (4%) with other neoplasms (P = .289). The median number of treatment cycles and time of exposure to bisphosphonates were 35 infusions and 39.3 months for patients with ONJ compared with 15 infusions (P < .001) and 19 months (P = .001), respectively, for patients with no ONJ. The incidence of ONJ increased with time to exposure from 1.5% among patients treated for 4 to 12 months to 7.7% for treatment of 37 to 48 months. The cumulative hazard was significantly higher with zoledronic acid compared with pamidronate alone or pamidronate and zoledronic acid sequentially (P < .001). All but two patients with ONJ had a history of dental procedures within the last year or use of dentures. CONCLUSION: The use of bisphosphonates seems to be associated with the development of ONJ. Length of exposure seems to be the most important risk factor for this complication. The type of bisphosphonate may play a role and previous dental procedures may be a precipitating factor. PMID- 16314621 TI - Ototoxicity in children receiving platinum chemotherapy: underestimating a commonly occurring toxicity that may influence academic and social development. AB - PURPOSE: To describe the frequency and severity of ototoxicity in a series of pediatric patients treated with platinum-based chemotherapy. PATIENTS AND METHODS: Serial audiologic evaluations were conducted for 67 patients aged 8 months to 23 years who received platinum-based chemotherapy. Audiologic data was analyzed to determine time to hearing-loss using American Speech-Language-Hearing Association (ASHA) criteria, and the effects of treatment and patient characteristics on the incidence and severity of ototoxicity. RESULTS: Bilateral decreases in hearing were seen in 61% of patients (median time to hearing loss, 135 days). Children treated for medulloblastoma, osteosarcoma, and neuroblastoma had greater incidence and severity of hearing loss. Agreement between the usually reported National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) and ASHA criteria was inadequate. CONCLUSION: Traditional reporting of toxicity data (CTCAE) has under-reported ototoxicity and minimized the significance of hearing loss in children. As pediatric patients experience improved survival, the effects and implications of high-frequency hearing loss with regard to academic achievement and speech and language development are important considerations, especially in patients younger than 5 years. PMID- 16314622 TI - Chemotherapy and cardiotoxicity in older breast cancer patients: a population based study. AB - PURPOSE: Adjuvant chemotherapy, especially with anthracyclines, is known to cause acute and chronic cardiotoxicity in breast cancer patients. We studied the cardiac effects of chemotherapy in a population-based sample of breast cancer patients aged > or = 65 years with long-term follow-up. PATIENTS AND METHODS: In the Surveillance, Epidemiology, and End Results (SEER)-Medicare database, we analyzed treatments and outcomes among women > or = 65 years of age who were diagnosed with stage I to III breast cancer from January 1, 1992 to December 31, 1999. Propensity scores were used to control for baseline heart disease (HD) and other known predictors of chemotherapy, and Cox proportional hazards models were used to estimate the risk of cardiomyopathy (CM), congestive heart failure (CHF), and HD after chemotherapy. RESULTS: Of 31,748 women with stage I to III breast cancer, 5,575 (18%) received chemotherapy. Chemotherapy was associated with younger age, fewer comorbidities, hormone receptor negativity, multiple primary tumors, and advanced disease. Patients who received chemotherapy were less likely than other patients to have pre-existing HD (45% v 55%, respectively; P < .001). The hazard ratios for CM, CHF, and HD for patients treated with doxorubicin (DOX) compared with patients who received no chemotherapy were 2.48 (95% CI, 2.10 to 2.93), 1.38 (95% CI, 1.25 to 1.52), and 1.35 (95% CI, 1.26 to 1.44), respectively. The relative risk of cardiotoxicity among patients who received DOX compared with untreated patients remained elevated 5 years after diagnosis. CONCLUSION: When baseline HD was taken into account, chemotherapy, especially with anthracyclines, was associated with a substantially increased risk of CM. As the number of long-term survivors grows, identifying and minimizing the late effects of treatment will become increasingly important. PMID- 16314623 TI - Allele imbalance, or loss of heterozygosity, in normal breast epithelium of sporadic breast cancer cases and BRCA1 gene mutation carriers is increased compared with reduction mammoplasty tissues. AB - PURPOSE: Normal-appearing breast epithelium can contain genetic abnormalities, including allele imbalance (AI), also referred to as loss of heterozygosity. Whether abnormalities are associated with cancer or cancer risk is unknown. PATIENTS AND METHODS: We performed a miniallelotype, using 20 microsatellites, on each of 460 histologically normal, microdissected breast terminal ducto-lobular units (TDLUs) from three groups of women: sporadic breast cancer patients (SP; n = 18), BRCA1 gene mutation carriers (BRCA1; n = 16), and controls undergoing reduction mammoplasty (RM; n = 18). We analyzed the results using Fisher's exact tests, logistic regression, and generalized estimating equations. RESULTS: AI was increased three-fold in SP and BRCA1 groups compared with RM. Both the number of TDLUs with AI increased (eight [5%] of 162 in the RM group compared with 24 [15%] of 162 in the SP and 22 [16%] of 136 in the BRCA1 groups; P = .0150), and the proportion of patients with AI increased (five [28%] of 18 in the RM group compared with 15 [83%] of 18 in the SP and 13 [81%] of 16 in the BRCA1 groups; P = .0007). The adjusted odds ratios (OR) for AI in TDLU increased in SP (OR = 15.5) and BRCA1 (OR = 13.7) patients compared with RM (P = .0025). This result was particularly evident on chromosome 17q (P = .0393), where more AI was seen in BRCA1 (OR = 12.4) than in SP (OR = 4.9) patients or RM controls. CONCLUSION: Increased prevalence of AI in normal-appearing epithelium is associated with breast cancer and increased breast cancer risk. The increased prevalence may reflect dysregulation, even in normal-appearing epithelium, of genomic processes contributing to cancer development. The clinical significance of genetic alterations in the subset of controls remains to be determined. PMID- 16314624 TI - Temporal and geographic variation in the use of hematopoietic growth factors in older women receiving breast cancer chemotherapy: findings from a large population-based cohort. AB - PURPOSE: Hematopoietic growth factors have played a major role in preventing infection and shortening the duration of neutropenia in patients receiving cancer chemotherapy. Little information is available on how these growth factors are used in patients with cancer outside the clinical trial setting. We performed descriptive and exploratory analyses on the patterns and correlates of the use of hematopoietic growth factors in community-dwelling elderly patients. PATIENTS AND METHODS: We identified 5,843 women from the Surveillance, Epidemiology, and End Results (SEER)-Medicare-linked data cohorts who were diagnosed with breast cancer at age 65 or older in 1992 to 1999 from the 11 SEER areas and received chemotherapy. RESULTS: Overall, 17.3% of the elderly women with breast cancer chemotherapy received filgrastim and 6.8% received epoetin. The use of the growth factors increased significantly over time from 1992 to 1999 (P < .001 for trend). Compared with patients diagnosed in 1992 to 1994, patients diagnosed in 1998 to 1999 were more than five times and 65 times more likely to receive filgrastim and epoetin, respectively, after controlling for other factors such as age and comorbidity. There also was substantial geographic variation in the use of hematopoietic growth factors, ranging from 10.6% in Seattle to 22.9% in Atlanta. Significant predictors of growth factors included patient age, race, tumor stage, and comorbidity. CONCLUSION: There were substantial temporal and geographic variations in the use of hematopoietic growth factors among patients receiving chemotherapy for breast cancer. The nationwide and population-based Medicare claims provide potential for examining the effectiveness, medical costs, and cost effectiveness of hematopoietic growth factors in the community. PMID- 16314625 TI - Prophylactic oophorectomy reduces breast cancer penetrance during prospective, long-term follow-up of BRCA1 mutation carriers. AB - PURPOSE: Breast cancer penetrance estimates in BRCA1 mutation carriers have varied from 40% to 85%; this heterogeneity has been attributed to variations in risk among different study populations. No study has taken oophorectomy status into account in estimating penetrance. Because prophylactic oophorectomy reduces breast cancer risk by approximately 50%, we hypothesized that population differences in oophorectomy prevalence might significantly influence breast cancer penetrance estimates. METHODS: Females from multiple-case breast/ovarian cancer families that segregate deleterious BRCA1 mutations were observed prospectively for breast cancer incidence and oophorectomy. RESULTS: Within this cohort, 33 cases of breast cancer developed in 98 women with deleterious BRCA1 mutations during follow-up, yielding an estimated cumulative lifetime breast cancer risk of 80%. This estimate increased to 94% when the study participants were censored at the time of oophorectomy. Six of the 33 mutation-positive women who underwent oophorectomy during follow-up developed breast cancer, compared with 27 of 65 mutation carriers with intact ovaries (hazard ratio = 0.38; 95% CI, 0.15 to 0.97). Estimates of absolute breast cancer risk demonstrated that the protective effect of oophorectomy was strongest among women who were premenopausal at the time of surgery. When surgical status was ignored, the strong protective effect of oophorectomy, coupled with the high prevalence of the procedure in these families, led to a significantly lower estimate of the breast cancer penetrance in BRCA1 mutation carriers. CONCLUSION: Differing rates of oophorectomy likely represent an underappreciated basis for a portion of the heterogeneity in estimated breast cancer penetrance described in BRCA mutation carriers, particularly mutation carriers from extensively affected, multiple-case families. PMID- 16314626 TI - Phase III randomized trial of cisplatin plus placebo compared with cisplatin plus cetuximab in metastatic/recurrent head and neck cancer: an Eastern Cooperative Oncology Group study. AB - PURPOSE: Therapy of recurrent/metastatic squamous cell carcinoma of the head and neck results in median progression-free survival (PFS) of 2 months. These cancers are rich in epidermal growth factor receptor (EGFR). We wished to determine whether the addition of cetuximab, which inhibits activation of EGFR, would improve PFS. PATIENTS AND METHODS: Patients with recurrent/metastatic squamous cell carcinoma of the head and neck were randomly assigned to receive cisplatin every 4 weeks, with weekly cetuximab (arm A) or placebo (arm B). Tumor tissue was assayed for EGFR expression by immunohistochemistry. The primary end point was PFS. Secondary end points of interest were response rate, toxicity, overall survival, and correlation of EGFR with clinical end points. RESULTS: There were 117 analyzable patients enrolled. Median PFS was 2.7 months for arm B and 4.2 months for arm A. The hazard ratio for progression of arm A to arm B was 0.78 (95% CI, 0.54 to 1.12). Median overall survival was 8.0 months for arm B and 9.2 months for arm A (P = .21). The hazard ratio for survival by skin toxicity in cetuximab-treated patients was 0.42 (95% CI, 0.21 to 0.86). Objective response rate was 26% [corrected] for arm A and 10% [corrected] for arm B (P = .03). Enhancement of response was greater for patients with EGFR staining present in less than 80% of cells. CONCLUSION: Addition of cetuximab to cisplatin significantly improves response rate. There was a survival advantage for the development of rash. Progression-free and overall survival were not significantly improved by the addition of cetuximab in this study. PMID- 16314627 TI - Phase III study of fluorouracil, leucovorin, and levamisole in high-risk stage II and III colon cancer: final report of Intergroup 0089. AB - PURPOSE: In 1990, fluorouracil (FU) plus levamisole for 1 year became standard adjuvant treatment for patients with high-risk stages II and III colon cancer. Intergroup (INT) 0089 assessed the relative contributions of leucovorin and levamisole in such patients. PATIENTS AND METHODS: From 1988 to 1992, 3,794 patients were randomly assigned. Experimental treatment consisted of one of three chemotherapy regimens: the low-dose leucovorin plus FU (Mayo Clinic; LDLV) regimen, the high-dose leucovorin plus FU (Roswell Park; HDLV) regimen, and the low-dose leucovorin plus levamisole plus FU (LDLV plus LEV) regimen, each administered for 30 to 32 weeks. The control arm was levamisole plus FU (LEV) for 1 year. RESULTS: After a median follow-up of 10 years, of 3,561 eligible patients, 1,691 (47%) have died and 1,330 (37%) have experienced disease recurrence; 137 (10%) of those experiencing recurrence are still alive. A total of 481 patients (13%) died without evidence of recurrence, and 1,723 (48%) are alive and disease free. Although there were toxicity differences among the four arms, none was statistically superior in disease-free or overall survival. CONCLUSION: The 6- to 8-month regimens of LDLV and HDLV without levamisole used in this trial, rather than the previous standard regimen of 12 months of LEV, have become widely used. INT-0089 has long-term follow-up of the largest clinical trial of patients with high-risk colon cancer, documenting not only the durability of the treatment effects, but also the natural history of patients with high-risk colon cancer, and analyses of treatment based on age, race, and comorbid conditions such as obesity, diabetes, and second primary cancers. PMID- 16314628 TI - Phase I study of capecitabine with concomitant radiotherapy for patients with locally advanced pancreatic cancer: expression analysis of genes related to outcome. AB - PURPOSE: To establish the feasibility of capecitabine with concurrent radiotherapy (XRT) in patients with locally advanced (LA) pancreatic cancer and evaluate the effect of XRT on thymidine phosphorylase (TP), dihydropyrimidine dehydrogenase (DPD), and tumor necrosis factor-alpha (TNF-alpha). PATIENTS AND METHODS: Fifteen patients with LA pancreatic cancer received three-dimensional conformal XRT to a dose of 50.4 Gy with capecitabine at escalating doses from 600 to 1,250 mg/m2 bid (Monday through Friday). Following chemo-XRT, stable and responding patients were treated with capecitabine 2,000 mg/m2 orally bid for 14 days every 21 days. Tumor specimens were procured with endoscopic ultrasound guided fine-needle aspiration 1 week before and 2 weeks after chemo-XRT to evaluate TP, DPD, and TNF-alpha mRNA levels. RESULTS: Dose-limiting grade 3 diarrhea was observed in two of six patients treated at a capecitabine dose of 1,000 mg/m2 with XRT. Three patients (20%) achieved partial response. Mean percent difference in TP pre- and post-XRT was 119.2% (P = .1934). There was no significant differences in mean TNF-alpha, or DPD levels pre- and post-XRT (P = .1934 and .4922, respectively). TP and TNF-alpha levels were not significantly correlated both at pre- and post-XRT (P = .670 and P < .154, respectively). Median value of TP:DPD ratios at baseline was 2.65 (range, 0.36 to 11.08). No association between TP:DPD ratio and efficacy of capecitabine or severity of toxicities was identified. CONCLUSION: The recommended dose for phase II evaluation is capecitabine 800 mg/m2 bid (Monday through Friday) with concurrent XRT. This approach offers an easy alternative to intravenous fluorouracil as a radiosensitizer in these patients. Role of TP and TP:DPD ratio warrants further investigation in a larger clinical trial. PMID- 16314629 TI - Adverse effects of preoperative radiation therapy for rectal cancer: long-term follow-up of the Swedish Rectal Cancer Trial. AB - PURPOSE: To analyze the occurrence of subacute and late adverse effects in patients treated with preoperative irradiation for rectal cancer. PATIENTS AND METHODS: The study population included 1,147 patients randomly assigned to preoperative radiation therapy or surgery alone in the Swedish Rectal Cancer Trial conducted 1987 through 1990. Patient data were matched against the Swedish Hospital Discharge Register to identify patients admitted to hospital after the primary treatment of the rectal cancer. Patients with known residual disease were excluded, and patients with a recurrence were censored 3 months before the date of recurrence. Relative risks (RR) with 95% CIs were calculated. RESULTS: Irradiated patients were at increased risk of admissions during the first 6 months from the primary treatment (RR = 1.64; 95% CI, 1.21 to 2.22); these were mainly for gastrointestinal diagnoses. Overall, the two groups showed no difference in the risk of admissions more than 6 months from the primary treatment (RR = 0.95; 95% CI, 0.80 to 1.12). Regarding specific diagnoses, however, RRs were increased for admissions later than 6 months from the primary treatment in irradiated patients for unspecified infections, bowel obstruction, abdominal pain, and nausea. CONCLUSION: Gastrointestinal disorders, resulting in hospital admissions, seem to be the most common adverse effect of short-course preoperative radiation therapy in patients with rectal cancer. Bowel obstruction was the diagnosis of potentially greatest importance, which was more frequent in irradiated than in nonirradiated patients. PMID- 16314630 TI - Colon cancer survival is associated with decreasing ratio of metastatic to examined lymph nodes. AB - PURPOSE: Colorectal cancer is the second leading cause of cancer deaths in the United States, with poor survival predicted by regional lymph node (LN) metastasis. The impact of LN ratio (LNR) on survival is unknown in this disease. PATIENTS AND METHODS: We analyzed data from Intergroup trial 0089 of adjuvant chemotherapy for stage II and III patients with colon cancer, in which all patients received fluorouracil-based therapy. Survival was similar for all arms of the study, allowing us to evaluate all patients together. End points included overall survival (OS), cancer-specific survival (CSS), and disease-free survival (DFS). Multivariate analyses were performed on all patients and on groups according to LNR quartiles (LNR: < 0.05, 0.05 to 0.19, 0.2 to 0.39, and 0.4 to 1.0). Covariates included in the models were age, sex, tumor stage, grade, histology, number of positive LNs, number of LNs removed, and LNR. RESULTS: The median age was 63.7 years, and the median number of LNs removed was 11. In the multivariate analysis, LNR was a significant factor for OS, DFS, and CSS in patients with 10 to 15 LN and more than 15 LN removed but not for patients with less than 10 LN removed. Using quartiles, LNR maintained its significance for all three end points when patients were grouped by node status. CONCLUSION: After curative resection for colorectal cancer, the LNR is an important prognostic factor and should be used in stratification schemes for future clinical trials investigating adjuvant treatments. PMID- 16314631 TI - Recent chemotherapy reduces the sensitivity of [18F]fluorodeoxyglucose positron emission tomography in the detection of colorectal metastases. AB - PURPOSE: [18F]2-fluoro-2-deoxyglucose (FDG) -positron emission tomography (PET) has become a useful tool in the assessment of patients with colorectal cancer. Patients often undergo chemotherapy as treatment for primary or metastatic colorectal malignancy. Because cytotoxic chemotherapy may decrease the cellular metabolic activity of tumor, we assessed the effects of chemotherapy on PET imaging. PATIENTS AND METHODS: This is a prospective study examining detection of hepatic colorectal metastases by FDG-PET as related to use of chemotherapy. Pathologic analysis of the liver resection specimens was used as gold standard. RESULTS: There was significantly decreased tumor FDG uptake (as measured by the maximal standardized uptake value) in patients treated preoperatively with chemotherapy, resulting in less efficient detection of cancerous lesions. One biologic basis of this change in accuracy of PET was a significant decrease in the activity of the key glycolytic enzyme hexokinase in tumors from patients treated with chemotherapy. CONCLUSION: These results indicate that FDG-PET scanning should be interpreted in the context of concurrent cytotoxic therapy. FDG-PET scanning results may also be useful in assessment of response to such cytotoxic therapies. PMID- 16314632 TI - Ixabepilone (epothilone B analogue BMS-247550) is active in chemotherapy-naive patients with hormone-refractory prostate cancer: a Southwest Oncology Group trial S0111. AB - PURPOSE: The epothilones are a new class of tubulin-polymerizing agents with activity in taxane-sensitive and resistant tumor models. We evaluated ixabepilone (BMS-247550) in patients with metastatic hormone-refractory prostate cancer (HRPC). METHODS: Eligible patients had chemotherapy-naive metastatic HRPC, a Zubrod performance status of 0 to 2, and adequate organ function. All patients received BMS-247550 at 40 mg/m2 over 3 hours every 3 weeks. The primary end point was proportion of patients achieving a prostate-specific antigen (PSA) response. RESULTS: Forty-eight patients with metastatic HRPC were registered. Forty-two patients were eligible, with a median age of 73 years and a median PSA level of 111 ng/mL; 78% had bone-only or bone and soft tissue metastases, and 88% had objective radiologic disease progression at registration. Grade 3 and 4 adverse events (AEs) occurred in 16 and three patients, respectively. All grade 4 toxicities were neutropenia or leukopenia. The most frequent grade 3 AEs were neuropathy (eight patients), hematologic toxicity (seven patients), flu-like symptoms, and infection (five patients each). There were no grade 3/4 thrombocytopenia or grade 5 AEs. There were 14 confirmed PSA responses (33%; 95% CI, 20% to 50%); 72% of PSA responders had declines greater than 80%, and two patients achieved an undetectable PSA. The estimated median progression-free survival is 6 months (95% CI, 4 to 8 months), and the median survival is 18 months (95% CI, 13 to 24 months). CONCLUSION: Ixabepilone has demonstrated activity in patients with chemotherapy-naive metastatic HRPC. Major toxicities were neutropenia and neuropathy. Further testing to define its activity relative to standard therapy is warranted. PMID- 16314633 TI - Factors affecting older african american women's decisions to join the PLCO Cancer Screening Trial. AB - PURPOSE: The purpose of this study is to describe the factors associated with the decisions of older African American women to join the PLCO (Prostate, Lung, Colorectal and Ovarian) Cancer Screening Trial when recruited. METHODS: African American women between ages 55 and 74 years who were never diagnosed with a PLCO cancer were eligible for our study. Two methods of recruitment were used. First, mailings were sent to a random sample of women describing the PLCO followed by a telephone call to determine interest in the PLCO. If women were not interested in PLCO but consented to participate in our study, they were interviewed immediately. Second, we followed up with African American women who responded to mass mailings sent out before the start of our study by the Pittsburgh PLCO office. Women completed an interview about their cancer and clinical trial knowledge, attitudes, beliefs, and behaviors. The responses of women who joined the PLCO Trial are contrasted with the responses of women who did not join. RESULTS: Numerous factors were associated with the decision of older African American women to join the PLCO, including perceptions of cancer prevention and detection, the experience of having a loved one with cancer, knowledge of and experience with clinical trials, and beliefs regarding the benefits and risks of clinical trial participation. CONCLUSION: Minority recruitment to cancer clinical trials could be increased by designing interventions focused on individual, organizational, and community needs. PMID- 16314634 TI - Phase II trial of high-dose conformal radiation therapy with concurrent hepatic artery floxuridine for unresectable intrahepatic malignancies. AB - PURPOSE: A phase II trial was conducted to determine if high-dose radiation with concurrent hepatic arterial floxuridine would improve survival in patients with unresectable intrahepatic malignancies. PATIENTS AND METHODS: Three-dimensional conformal high-dose radiation therapy was delivered concurrently with hepatic arterial floxuridine in 128 patients. The radiation dose was based on a normal tissue complication probability model and subjected the patient to an estimated maximum risk of radiation-induced liver disease of 10% to 15%. The study design provided more than 80% power to detect a two-fold increase in median survival compared with historical controls at a 5% significance level. RESULTS: The median radiation dose delivered was 60.75 Gy (1.5-Gy fractions bid). At a median follow up time of 16 months (26 months in patients who were alive) the median survival was 15.8 months (95% CI, 12.6 to 18.3 months), significantly longer than in the historical control. The actuarial 3-year survival was 17%. The total dose was the only significant predictor of survival. Primary hepatobiliary tumors had a significantly greater tendency to remain confined to the liver than did colorectal cancer metastases. Overall toxicity was acceptable, with 27 patients (21%) and 11 patients (9%) developing grade 3 and 4 toxicity, respectively, and one treatment-related death. CONCLUSION: The results suggest that, compared with historical controls, high-dose focal liver irradiation with hepatic artery floxuridine prolongs survival in patients with unresectable chemotherapy refractory metastatic colorectal cancer and primary hepatobiliary tumors. This provides a rationale for intensification of local therapy for unresectable hepatobiliary cancers and integration of this regimen with newer systemic therapy for patients with colorectal cancer. PMID- 16314635 TI - Early variations of circulating interleukin-6 and interleukin-10 levels during thoracic radiotherapy are predictive for radiation pneumonitis. AB - PURPOSE: To investigate variations of circulating serum levels of interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), and interleukin-10 (IL-10) during three-dimensional conformal radiation therapy (3D-CRT) in patients with non-small cell lung cancer and correlate these variations with the occurrence of radiation pneumonitis. PATIENTS AND METHODS: Ninety-six patients receiving 3D-CRT for stage I to III disease were evaluated prospectively. Circulating cytokine levels were determined before, every 2 weeks during, and at the end of treatment. Radiation pneumonitis was evaluated prospectively between 6 and 8 weeks after 3D-CRT. The predictive value of clinical, dosimetric, and biologic (cytokine levels) factors was evaluated both in univariate and multivariate analyses. RESULTS: Forty patients (44%) experienced score 1 or more radiation pneumonitis. No association was found between baseline cytokine levels and the risk of radiation pneumonitis. In the whole population, mean levels of TNFalpha, IL-6, and IL-10 remained stable during radiotherapy. IL-6 levels were significantly higher (P = .047) during 3D CRT in patients with radiation pneumonitis. In the multivariate analysis, covariations of IL-6 and IL-10 levels during the first 2 weeks of 3D-CRT were evidenced as independently predictive of radiation pneumonitis in this series (P = .011). CONCLUSION: Early variations of circulating IL-6 and IL-10 levels during 3D-CRT are significantly associated with the risk of radiation pneumonitis. Variations of circulating IL-6 and IL-10 levels during 3D-CRT may serve as independent predictive factors for this complication. PMID- 16314636 TI - Bexarotene and erlotinib for aerodigestive tract cancer. AB - PURPOSE: The epidermal growth factor receptor (EGFR) and cyclin D1 are overexpressed in lung carcinogenesis. The rexinoid, bexarotene, represses cyclin D1 and EGFR expression in vitro. It was hypothesized that combining bexarotene with the EGFR inhibitor, erlotinib, would augment clinical activity. PATIENTS AND METHODS: In vitro studies and a phase I clinical trial were performed. Twenty four patients with advanced aerodigestive tract cancers were enrolled; 79% had non-small-cell lung cancer (NSCLC). The primary objective was to determine the maximum-tolerated dose. Clinical activity was a secondary objective. RESULTS: Combining erlotinib with bexarotene enhanced growth suppression in vitro compared with each single-agent treatment. This cooperatively repressed cyclin D1 expression. Clinically, the most frequent toxicities were mild hypertriglyceridemia and skin rash. Two serious treatment-related adverse events occurred (creatine phosphokinase elevation attributed to antilipid therapy and a case of generalized pain). Five objective responses (four partial and one minor) were observed in NSCLC patients. Responses were observed in males and smokers. EGFR sequence analyses did not reveal activating mutations in tumors from assessable responding patients. Median time to progression was 2.0 months; overall survival time was 14.1 months; and 1-year survival rate was 73.8%. CONCLUSION: The recommended phase II doses are erlotinib 150 mg/d and bexarotene 400 mg/m2/d orally. These agents can be administered in combination at the recommended single-agent doses without added toxicity. Overall survival and clinical features of responding patients differ from prior reports of single agent erlotinib treatment. These findings are encouraging and warrant further investigation of this regimen. PMID- 16314637 TI - Wnt5a expression is associated with the tumor proliferation and the stromal vascular endothelial growth factor--an expression in non-small-cell lung cancer. AB - PURPOSE: The Wnt gene family encodes the multifunctional signaling glycoproteins. We performed the present study to investigate the clinical significance of Wnt5a expression in non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: One hundred twenty-three patients with NSCLC who had undergone resection were investigated. Real-time quantitative reverse transcriptase polymerase chain reaction was performed to evaluate the Wnt5a gene expression. Immunohistochemistry was performed to investigate the Wnt5a protein expression, the Ki-67 proliferation index, tumor angiogenesis, and the expression of beta catenin and vascular endothelial growth factor-A (VEGF-A). RESULTS: Wnt5a gene expression in squamous cell carcinoma was significantly higher than that in adenocarcinoma (P < .0001). There was a significant correlation between the normalized Wnt5a gene expression ratio and the intratumoral Wnt5a protein expression (r = 0.729; P < .0001). The intratumoral Wnt5a expression was significantly correlated with the Ki-67 proliferation index (r = 0.708; P < .0001). In contrast, no correlation was observed between the intratumoral Wnt5a expression and tumor angiogenesis. Furthermore, the intratumoral Wnt5a expression was significantly correlated with the stromal expression of beta-catenin (r = 0.729; P < .0001) and VEGF-A (r = 0.661; P < .0001). In addition, the stromal VEGF-A expression was also correlated with Ki-67 proliferation (r = 0.627; P < .0001). Cox regression analyses demonstrated Wnt5a status to be a significant prognostic factor for NSCLC patients (P = .0193), especially for patients with squamous cell carcinomas (P = .0491). CONCLUSION: The present study revealed that an overexpression of Wnt5a could produce more aggressive NSCLC, especially in squamous cell carcinomas, during tumor progression. PMID- 16314638 TI - Role of chemotherapy and the receptor tyrosine kinases KIT, PDGFRalpha, PDGFRbeta, and Met in large-cell neuroendocrine carcinoma of the lung. AB - PURPOSE: Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is a relatively uncommon, high-grade neuroendocrine tumor sharing several features with small cell lung carcinoma (SCLC) but currently considered as a variant of non-SCLC and accordingly treated with poor results. Little is known about the optimal therapy of LCNEC and the possible therapeutic molecular targets. PATIENTS AND METHODS: We reviewed 83 patients with pure pulmonary LCNEC to investigate their clinicopathologic features, therapeutic strategy, and immunohistochemical expression and the mutational status of the receptor tyrosine kinases (RTKs) KIT, PDGFRalpha, PDGFRbeta, and Met. RESULTS: LCNEC histology predicted a dismal outcome (overall median survival, 17 months) even in stage I patients (5-year survival rate, 33%). LCNEC strongly expressed RTKs (KIT in 62.7% of patients, PDGFRalpha in 60.2%, PDGFRbeta in 81.9%, and Met in 47%), but no mutations were detected in the exons encoding for the relevant juxtamembrane domains. Tumor stage and size (> or = 3 cm) and Met expression were significantly correlated with survival. At univariate and multivariate analysis, SCLC-based chemotherapy (platinum-etoposide) was the most important variable correlating with survival, both in the adjuvant and metastatic settings (P < .0001). CONCLUSION: Pulmonary LCNEC represents an aggressive tumor requiring multimodal treatment even for resectable stage I disease, and LCNEC seems to respond to adjuvant platinum etoposide-based chemotherapy. Patients who received this therapy had the best survival rate. Despite our failure in finding mutational events in the tested RTKs, the strong expression of KIT, PDGFRalpha, PDGFRbeta, and Met in tumor cells suggests an important role of these RTKs in LCNEC, and these RTKs seem to be attractive therapeutic targets. PMID- 16314639 TI - Logistic regression model to distinguish between the benign and malignant adnexal mass before surgery: a multicenter study by the International Ovarian Tumor Analysis Group. AB - PURPOSE: To collect data for the development of a more universally useful logistic regression model to distinguish between a malignant and benign adnexal tumor before surgery. PATIENTS AND METHODS: Patients had at least one persistent mass. More than 50 clinical and sonographic end points were defined and recorded for analysis. The outcome measure was the histologic classification of excised tissues as malignant or benign. RESULTS: Data from 1,066 patients recruited from nine European centers were included in the analysis; 800 patients (75%) had benign tumors and 266 (25%) had malignant tumors. The most useful independent prognostic variables for the logistic regression model were as follows: (1) personal history of ovarian cancer, (2) hormonal therapy, (3) age, (4) maximum diameter of lesion, (5) pain, (6) ascites, (7) blood flow within a solid papillary projection, (8) presence of an entirely solid tumor, (9) maximal diameter of solid component, (10) irregular internal cyst walls, (11) acoustic shadows, and (12) a color score of intratumoral blood flow. The model containing all 12 variables (M1) gave an area under the receiver operating characteristic curve of 0.95 for the development data set (n = 754 patients). The corresponding value for the test data set (n = 312 patients) was 0.94; and a probability cutoff value of .10 gave a sensitivity of 93% and a specificity of 76%. CONCLUSION: Because the model was constructed from multicenter data, it is more likely to be generally applicable. The effectiveness of the model will be tested prospectively at different centers. PMID- 16314640 TI - Does aggressive surgery only benefit patients with less advanced ovarian cancer? Results from an international comparison within the SCOTROC-1 Trial. AB - PURPOSE: Studies indicate that ovarian cancer patients who have been optimally debulked survive longer. Although chemotherapy has been variable, they have defined standards of care. Additionally, it is suggested that patients from the United Kingdom (UK) have inferior survival compared with some other countries. We explored this within the context of a large, international, prospective, randomized trial of first-line chemotherapy in advanced ovarian cancer (docetaxel carboplatin v paclitaxel-carboplatin; SCOTROC-1). The Scottish Randomised Trial in Ovarian Cancer surgical study is a prospective observational study examining the impact on progression-free survival (PFS) of cytoreductive surgery and international variations in surgical practice. PATIENTS AND METHODS: One thousand seventy-seven patients were recruited (UK, n = 689; Europe, United States, and Australasia, n = 388). Surgical data were available for 889 patients. These data were analyzed within a Cox model. RESULTS: There were three main observations. First, more extensive surgery was performed in non-UK patients, who were more likely to be optimally debulked (< or = 2 cm residual disease) than UK patients [corrected] (71.3% v 58.4%, respectively; P < .001). Second, optimal debulking was associated with increased PFS mainly for patients with less extensive disease at the outset (test for interaction, P = .003). Third, UK patients with no visible residual disease had a less favorable PFS compared with patients recruited from non-UK centers who were similarly debulked (hazard ratio = 1.85; 95% CI, 1.16 to 2.97; P = .010). This observation seems to be related to surgical practice, primarily lymphadenectomy. CONCLUSION: Increased PFS associated with optimal surgery is limited to patients with less advanced disease, arguing for case selection rather than aggressive debulking in all patients irrespective of disease extent. Lymphadenectomy may have beneficial effects on PFS in optimally debulked patients. PMID- 16314641 TI - Genetic testing in pheochromocytoma or functional paraganglioma. AB - PURPOSE: To assess the yield and the clinical value of systematic screening of susceptibility genes for patients with pheochromocytoma (pheo) or functional paraganglioma (pgl). PATIENTS AND METHODS: We studied 314 patients with a pheo or a functional pgl, including 56 patients having a family history and/or a syndromic presentation and 258 patients having an apparently sporadic presentation. Clinical data and blood samples were collected, and all five major pheo-pgl susceptibility genes (RET, VHL, SDHB, SDHD, and SDHC) were screened. Neurofibromatosis type 1 was diagnosed from phenotypic criteria. RESULTS: We have identified 86 patients (27.4%) with a hereditary tumor. Among the 56 patients with a family/syndromic presentation, 13 have had neurofibromatosis type 1, and germline mutations on the VHL, RET, SDHD, and SDHB genes were present in 16, 15, nine, and three patients, respectively. Among the 258 patients with an apparently sporadic presentation, 30 (11.6%) had a germline mutation (18 patients on SDHB, nine patients on VHL, two patients on SDHD, and one patient on RET). Mutation carriers were younger and more frequently had bilateral or extra-adrenal tumors. In patients with an SDHB mutation, the tumors were larger, more frequently extra adrenal, and malignant. CONCLUSION: Genetic testing oriented by family/sporadic presentation should be proposed to all patients with pheo or functional pgl. We suggest an algorithm that would allow the confirmation of suspected inherited disease as well as the diagnosis of unexpected inherited disease. PMID- 16314642 TI - Outcomes of children with intermediate-risk neuroblastoma after treatment stratified by MYCN status and tumor cell ploidy. AB - PURPOSE: The goal of Pediatric Oncology Group 9243 was to improve outcomes for children with intermediate-risk neuroblastoma (NB). PATIENTS AND METHODS: Patients were assigned to treatments on the basis of age, tumor MYCN status, and tumor cell ploidy. Children in the less intensive arm A received cyclophosphamide/doxorubicin and surgery. Patients not in complete remission postoperatively were treated with cisplatin/etoposide, cyclophosphamide/doxorubicin, and additional surgery. Patients with less favorable features were assigned to arm B, which consisted of carboplatin, etoposide, ifosfamide, and surgery. Survival rates were determined using an intent-to-treat approach. RESULTS: For arm-A patients, the 6-year event-free survival (EFS) was 86% with an SE of 3%. For arm-B patients, the 6-year EFS was 46% with an SE of 7%. MYCN status was the only statistically significant prognostic variable. Among patients whose tumors were MYCN nonamplified, a trend toward improved EFS was seen in children with hyperdiploid versus diploid tumors. However, many of these children responded well to salvage therapy, and overall survival rates did not differ on the basis of ploidy. Six-year EFS rates for arm B were patients with MYCN nonamplified, hyperdiploid tumors, 86% with an SE of 3%; patients with MYCN nonamplified, diploid tumors, 74% with an SE of 10%; patients with MYCN-amplified, hyperdiploid tumors, 46% with an SE of 15%; and patients with MYCN-amplified, diploid tumors, 22% with an SE of 10%. CONCLUSION: Outcomes for patients with MYCN-nonamplified, hyperdiploid tumors were excellent. Therapy reductions for these patients merit study. A trend toward less favorable outcomes for patients with MYCN-nonamplified, diploid tumors was observed; more children may need to be evaluated before therapy is reduced for this subgroup. For patients with MYCN-amplified tumors, new strategies are needed. PMID- 16314643 TI - [18F]Fluorodeoxyglucose positron emission tomography predicts outcome for Ewing sarcoma family of tumors. AB - PURPOSE: Response to neoadjuvant chemotherapy is a significant prognostic factor for the Ewing sarcoma family of tumors (ESFTs). [18F]fluorodeoxyglucose (FDG) positron emission tomography (PET) is a noninvasive imaging modality that accurately predicts histopathologic response in several malignancies. To determine the prognostic value of FDG PET response for progression-free survival (PFS) in ESFTs, we reviewed the University of Washington Medical Center experience. PATIENTS AND METHODS: Thirty-six patients with ESFTs were evaluated by FDG PET. All patients received neoadjuvant and adjuvant chemotherapy. FDG PET standard uptake values before (SUV1) and after (SUV2) chemotherapy were analyzed and correlated with chemotherapy response, as assessed by histopathology in surgically excised tumors. Thirty-four patients had both SUV1 and SUV2. RESULTS: The mean SUV1, SUV2, and ratio of SUV2 to SUV1 (SUV2:1) were 7.9 (range, 2.3 to 32.8), 2.1 (range, 0 to 4.3), and 0.36 (range, 0.00 to 1.00), respectively. Good FDG PET response was defined as SUV2 less than 2.5 or SUV2:1 < or = 0.5. FDG PET response by SUV2 or SUV2:1 was concordant with histologic response in 68% and 69% of patients, respectively. SUV2 was associated with outcome (4-year PFS 72% for SUV2 < 2.5 v 27% for SUV2 > or = 2.5, P = .01 for all patients; 80% for SUV2 < 2.5 v 33% for SUV2 > or = 2.5, P = .036 for localized at diagnosis patients). SUV2:1 < or = 0.5 was not predictive of PFS. CONCLUSION: FDG PET imaging of ESFTs correlates with histologic response to neoadjuvant chemotherapy. SUV2 less than 2.5 is predictive of PFS independent of initial disease stage. PMID- 16314644 TI - Phase I study of the sequential combination of interleukin-12 and interferon alfa 2b in advanced cancer: evidence for modulation of interferon signaling pathways by interleukin-12. AB - PURPOSE: To evaluate the safety of sequentially administered recombinant (r) human (h) interleukin-12 (IL-12) and interferon alfa-2b (IFN-alpha-2b) in patients with advanced cancer and to determine the effects of endogenously produced IFN-gamma on Janus kinase-signal transducer and activator of transcription (Jak-STAT) signal transduction in patient peripheral-blood mononuclear cells (PBMCs). PATIENTS AND METHODS: Forty-nine patients with metastatic cancer received rhIL-12 on day 1 and IFN-alpha-2b on days 2 to 6 of either a 14-day (n = 43) or a 7-day treatment cycle (n = 6). rhIL-12 was initially administered subcutaneously at a dose of 100 ng/kg, whereas IFN-alpha 2b was escalated from 1 to 10 million units (MU) in cohorts of three patients (1, 3, 5, 7, or 10 MU). rhIL-12 was subsequently administered intravenously (IV) in escalating doses (100 to 500 ng/kg) to achieve greater IFN-gamma production. Peripheral blood was drawn for measurement of plasma IFN-gamma and the induction of Jak-STAT signal transduction in PBMCs. RESULTS: No IL-12-or IFN-alpha-related dose-limiting toxicities were observed. There were no responses in 41 assessable patients. Five patients exhibited stable disease lasting 6 months or longer while on therapy. Optimal induction of IFN-gamma by IL-12 occurred after an IV dose of 250 ng/kg. Patient PBMCs exhibited increased levels of STAT1 after IL-12 administration. The peak level of IFN-gamma achieved with IL-12 therapy correlated with the peak level of intracellular STAT1 in patient PBMCs (r = 0.38, P = .021). CONCLUSION: The combination of rhIL-12 and IFN-alpha-2b can be administered sequentially with minimal toxicity. IV administration of rhIL-12 modulates IFN-alpha-induced Jak-STAT signal transduction in patient PBMCs. PMID- 16314645 TI - Genomic and protein expression profiling identifies CDK6 as novel independent prognostic marker in medulloblastoma. AB - PURPOSE: Medulloblastoma is the most common malignant brain tumor in children. Despite multimodal aggressive treatment, nearly half of the patients die as a result of this tumor. Identification of molecular markers for prognosis and development of novel pathogenesis-based therapies depends crucially on a better understanding of medulloblastoma pathomechanisms. PATIENTS AND METHODS: We performed genome-wide analysis of DNA copy number imbalances in 47 medulloblastomas using comparative genomic hybridization to large insert DNA microarrays (matrix-CGH). The expression of selected candidate genes identified by matrix-CGH was analyzed immunohistochemically on tissue microarrays representing medulloblastomas from 189 clinically well-documented patients. To identify novel prognostic markers, genomic findings and protein expression data were correlated to patient survival. RESULTS: Matrix-CGH analysis revealed frequent DNA copy number alterations of several novel candidate regions. Among these, gains at 17q23.2-qter (P < .01) and losses at 17p13.1 to 17p13.3 (P = .04) were significantly correlated to poor prognosis. Within 17q23.2-qter and 7q21.2, two of the most frequently gained chromosomal regions, confined amplicons were identified that contained the PPM1D and CDK6 genes, respectively. Immunohistochemistry revealed strong expression of PPM1D in 148 (88%) of 168 and CDK6 in 50 (30%) of 169 medulloblastomas. Overexpression of CDK6 correlated significantly with poor prognosis (P < .01) and represented an independent prognostic marker of overall survival on multivariate analysis (P = .02). CONCLUSION: We identified CDK6 as a novel molecular marker that can be determined by immunohistochemistry on routinely processed tissue specimens and may facilitate the prognostic assessment of medulloblastoma patients. Furthermore, increased protein-levels of PPM1D and CDK6 may link the TP53 and RB1 tumor suppressor pathways to medulloblastoma pathomechanisms. PMID- 16314646 TI - Efficacy of fractionated stereotactic reirradiation in recurrent gliomas: long term results in 172 patients treated in a single institution. AB - PURPOSE: To evaluate the efficacy of fractionated stereotactic radiotherapy (FSRT) performed as reirradiation in 172 patients with recurrent low- and high grade gliomas. PATIENTS AND METHODS: Between 1990 and 2004, 172 patients with recurrent gliomas were treated with FSRT as reirradiation in a single institution. Seventy-one patients suffered from WHO grade 2 gliomas. WHO grade 3 gliomas were diagnosed in 42 patients, and 59 patients were diagnosed with glioblastoma multiforme (GBM). The median time between primary radiotherapy and reirradiation was 10 months for GBM, 32 months for WHO grade 3 tumors, and 48 months for grade 2 astrocytomas. FSRT was performed with a median dose of 36 Gy in a median fractionation of 5 x 2 Gy/wk. RESULTS: Median overall survival after primary diagnosis was 21 months for patients with GBM, 50 months for patients with WHO grade 3 gliomas, and 111 months for patients with WHO grade 2 gliomas. Histologic grading was the strongest predictor for overall survival, together with the extent of neurosurgical resection and age at primary diagnosis. Median survival after reirradiation was 8 months for patients with GBM, 16 months for patients with grade 3 tumors, and 22 months for patients with low-grade gliomas. Only time to progression and histology were significant in influencing survival after reirradiation. Progression-free survival after FSRT was 5 months for GBM, 8 months for WHO grade 3 tumors, and 12 months for low-grade gliomas. CONCLUSION: FSRT is well tolerated and may be effective in patients with recurrent gliomas. Prospective studies are warranted for further evaluation. PMID- 16314647 TI - Phase II trial of radiosurgery for one to three newly diagnosed brain metastases from renal cell carcinoma, melanoma, and sarcoma: an Eastern Cooperative Oncology Group study (E 6397). AB - PURPOSE: Long-term brain metastases survivors are at risk for neurologic morbidity after whole-brain radiotherapy (WBRT). Retrospective radiosurgery (RS) reports found no survival difference when compared with WBRT. Before RS alone was evaluated with delayed WBRT in a phase III trial, the feasibility of RS alone was tested prospectively. PATIENTS AND METHODS: Patients with renal cell carcinoma, melanoma, or sarcoma; one to three brain metastases; and performance status of 0 to 2 were enrolled. Exclusion criteria were leptomeningeal disease; metastases in medulla, pons, or midbrain; or liver metastases. On the basis of tumor size, patients received 24, 18, or 15 Gy RS. At recurrence, management was discretionary. The primary end point was 3- and 6-month intracranial progression. RESULTS: Between July 1998 and August 2003, 36 patients were accrued; 31 were eligible. Median follow-up was 32.7 months and the median survival was 8.3 months (95% CI, 7.4 to 12.2). Three- and 6-month intracranial failure with RS alone was 25.8% and 48.3%. Failure within and outside the RS volume, when in-field and distant intracranial failures were scored independently, was 19.3% and 16.2% (3 months) and 32.2% and 32.2% (6 months), respectively. Approximately 38% of patients experienced death attributable to neurologic cause. There were three grade 3 toxicities related to RS. CONCLUSION: Intracranial failure rates without WBRT were 25.8% and 48.3% at 3 and 6 months, respectively. Delaying WBRT may be appropriate for some subgroups of patients with radioresistant tumors, but routine avoidance of WBRT should be approached judiciously. PMID- 16314648 TI - Costs and cost effectiveness of a health care provider-directed intervention to promote colorectal cancer screening among Veterans. AB - PURPOSE: Colorectal cancer screening is underused, particularly in the Veterans Affairs (VA) population. In a randomized controlled trial, a health care provider directed intervention that offered quarterly feedback to physicians on their patients' colorectal cancer screening rates led to a 9% increase in colorectal cancer screening rates among veterans. The objective of this secondary analysis was to assess the cost effectiveness of the colorectal cancer screening promotion intervention. METHODS: Providers in the intervention arm attended an educational workshop on colorectal cancer screening and received confidential feedback on individual and group-specific colorectal cancer screening rates. The primary end point was completion of colorectal cancer screening tests. Sensitivity analyses investigated cost-effectiveness estimates varying the data collection methods, costs of labor and technology, and the effectiveness of the intervention. RESULTS: Rates of colorectal cancer screening for the intervention versus control arms were 41.3% v 32.4%, respectively (P < .05). The incremental cost effectiveness ratio was dollar 978 per additional veteran screened based on feedback reports generated from manual review of records. However, if feedback reports could be generated from information technology systems, sensitivity analyses indicate that the cost-effectiveness estimate would decrease to dollar 196 per additional veteran screened. CONCLUSION: An intervention based on quarterly feedback reports to physicians improved colorectal cancer screening rates at a VA medical center. This intervention would be cost effective if relevant data could be generated by existing information technology systems. Our findings may have broad applicability because a 2005 Medicare initiative will provide the VA electronic medical record system as a free benefit to all US physicians. PMID- 16314649 TI - Health behaviors of cancer survivors: examining opportunities for cancer control intervention. AB - PURPOSE: A population-based investigation was conducted to examine the prevalence of health behaviors (smoking, alcohol use, physical activity, and cancer screening) of cancer survivors by age, time since diagnosis, and cancer site. Understanding health behaviors of survivors is imperative, as many survivors are living longer and are at risk for cancer recurrence, second cancers, and complications from treatment. METHODS: Using the National Health Interview Survey, this study examined the prevalence of smoking and alcohol use as well as whether cancer survivors (n = 7,384) are meeting current recommendations for physical activity and cancer screening compared with noncancer controls (n = 121,347). RESULTS: Cancer survivors are similar to controls with respect to smoking status and alcohol consumption after adjusting for group differences. However, younger survivors (18 to 40 years) are at greater risk for continued smoking than controls. Survivors are 9% more likely to meet physical activity recommendations compared with controls. chi2 tests indicate no significant differences in smoking, alcohol consumption, and physical activity by time since diagnosis, but differences by cancer site exist. Female cancer survivors are 34% and 36% more likely to meet mammogram and Papanicolaou smear screening recommendations, respectively, compared with controls. Similar screening patterns were found for prostate-specific antigen screening in men. CONCLUSION: This study provides benchmark approximations of the prevalence of risky health behaviors of survivors by time since diagnosis and cancer site. As part of the collective effort to reduce late effects of cancer treatment, oncologists may be in the best position to offer initial guidance for promoting healthy lifestyle behaviors among cancer survivors. PMID- 16314650 TI - Caveat medicus: consequences of federal investigations of marketing activities of pharmaceutical suppliers of prostate cancer drugs. AB - In the course of recent health care fraud investigations against TAP Pharmaceuticals (Lake Forest, IL) and AstraZeneca International (London, United Kingdom), each pled guilty to one violation of the Prescription Drug Marketing Act, settled claims related to alleged violations of the False Claims Act without admitting guilt, and paid fines, settlements for liabilities, and reimbursements of dollar 850 million and dollar 355 million, respectively. In a unique aspect of these cases, federal investigators brought criminal charges against 14 TAP employees and investigated the billing practices of several urologists. These investigations resulted in guilty pleas from both urologists and industry employees relative to the Prescription Drug Marketing Act or the False Claims Act and probationary sentences with payments of fines and restitution to the government for urologists who cooperated with federal investigations. One uncooperative urologist was found guilty of violating the Federal False Claims Act and sentenced to 6 months of home arrest, excluded from Medicare for 5 years, required to provide 600 hours of free medical care to indigent patients and patients covered by Medicare or Medicaid, and paid fines and restitution to the government. The cases against TAP and AstraZeneca have been followed by federal and state investigations of allegedly illegal marketing practices of other pharmaceutical firms and have resulted in negotiated settlements of dollar 3.8 billion and dollar 71.5 million, respectively. Believing that an Average Wholesale Price-based reimbursement system was an important driving factor for these marketing activities, Medicare has shifted to an Average Sales Price-based reimbursement system. This is expected to greatly impact the practice of outpatient oncology nationwide. PMID- 16314651 TI - Positive positron emission tomography scan in sarcoidosis and two challenging cases of metastatic cancer. CASE 1. Mediastinal sarcoidosis in a melanoma patient treated with interferon. PMID- 16314652 TI - Positive positron emission tomography scan in sarcoidosis and two challenging cases of metastatic cancer. CASE 2. Hepatocellular carcinoma with metastasis to the breast. PMID- 16314653 TI - Positive positron emission tomography scan in sarcoidosis and two challenging cases of metastatic cancer. CASE 3. Hoarseness caused by cancer metastasis to the vagus nerve. PMID- 16314654 TI - A Bayesian approach to a patient with a residual mass after treatment for non Hodgkin's lymphoma of the thyroid. PMID- 16314655 TI - Radiotherapy and Hodgkin's lymphoma. PMID- 16314656 TI - Heparanase activity and bone loss in postmenopausal breast cancer patients. PMID- 16314657 TI - Can the reliance of hormone receptor assays of surgical specimens be explained by the fluctuation of estrogen receptor, progesterone receptor, and HER-2 protein expression in tumor samples of premenopausal breast cancer patients? PMID- 16314658 TI - Circulating MYCN DNA predicts MYCN-amplification in neuroblastoma. PMID- 16314659 TI - After-rape among three populations in the Southwest: a time of mourning, a time for recovery. AB - Narrative analysis of open-ended interviews with 62 female survivors of rape from three populations in the Southwest (Native American, Mexican American, Anglo) uncovered commonalities and dissimilarities in women's description of their experience of afterrape (rape survival). Although all three groups reported experiences that confirm aspects of prior analyses of reactions to rape, the narrative analysis highlights variations in reactions to rape across the three groups. These variations, and more established commonalities, provide baseline material for strengthening primary and secondary interventions for women who have experienced sexual violence. PMID- 16314660 TI - Rape survivors' experiences with the legal and medical systems: do rape victim advocates make a difference? AB - This study used a naturalistic quasi-experimental design to examine whether rape survivors who had the assistance of rape victim advocates had more positive experiences with the legal and medical systems compared to those who did not work with advocates. Eighty-one survivors were interviewed in two urban hospitals about what services they received from legal and medical system personnel and how they were treated during these interactions. Survivors who had the assistance of an advocate were significantly more likely to have police reports taken and were less likely to be treated negatively by police officers. These women also reported less distress after their contact with the legal system. Similarly, survivors who worked with an advocate during their emergency department care received more medical services, including emergency contraception and sexually transmitted disease prophylaxis, reported significantly fewer negative interpersonal interactions with medical system personnel, and reported less distress from their medical contact experiences. PMID- 16314661 TI - An examination of the effectiveness of domestic violence coordinating councils. AB - The community response to domestic violence has been characterized by a lack of coordination across systems. Recent efforts have focused on bringing together stakeholders from an array of community sectors to promote a comprehensive response. Coordinating councils are commonly employed to encourage such a response, yet little is known about the extent to which they effectively promote community change. This study examined 41 councils in one state to better understand (a) the types of issues they addressed, (b) the extent to which they reported accomplishing their goals, and (c) the degree to which councils' climate and membership influenced the extent to which they achieved their goals within the criminal justice system and in other community sectors (e.g., health care, human service). PMID- 16314662 TI - A summary of personal protection order statutes in the United States. AB - A systematic analysis of statutes regarding personal protection orders (PPOs) was completed for all 50 states plus the District of Columbia to determine which states are most "victim friendly" in protecting women against domestic violence. Indicators used include compliance with the Violence Against Women Act (VAWA), the relationship between petitioner and respondent, ease of the PPO administrative process, and severity of punishment for violations. Results indicate that most states have PPO legislation that is consistent with VAWA; however, states differ dramatically in accessibility of orders for victims of violence. PMID- 16314663 TI - Women's use of force: voices of women arrested for domestic violence. AB - Following changes in law enforcement policies that encourage or mandate arrest of domestic violence offenders, a concomitant increase in women arrested and mandated to batterer treatment programs has resulted. Most research findings, however, suggest that heterosexual intimate violence is gendered, with abuse, power, and control wielded by men over their female partners, and that when women use violence, it is typically in self defense or for nonaggressive reasons. However, few studies have investigated the female batterer treatment programs and the context of the women's use of violence. Using qualitative data collected from observations of three female domestic violence offender programs, this article examines women's interpretations of their violent experiences. In addition, the findings raise policy-level questions about the appropriateness of such programs, weighing the costs and benefits of a criminal justice approach to women's use of force in intimate relationships. PMID- 16314664 TI - The search for quality. PMID- 16314665 TI - Presidential address of the American Orthopaedic Society for Sports Medicine: Education: commodity or responsibility? PMID- 16314667 TI - Suture anchors and tacks for shoulder surgery, part 1: biology and biomechanics. AB - The development and successful clinical application of suture anchors and tacks have revolutionized the surgeon's ability to secure soft tissues to bone via open or arthroscopic surgical techniques. When used carefully and with proper technique, these devices provide viable options for the repair and reconstruction of many intra-articular and extra-articular abnormalities in the shoulder, including rotator cuff tears, shoulder instability, and biceps lesions that require labrum repair or biceps tendon tenodesis. Like many technologies, however, the successful application of these devices requires an understanding of the biology and biomechanics that affect their use in the shoulder as well as knowledge of the factors that can affect subsequent clinical outcomes, including complications. PMID- 16314668 TI - Community-acquired methicillin-resistant staphylococcus aureus: an emerging problem in the athletic population. AB - Participants of contact sports are at risk for outbreaks of skin and soft tissue infection. Causes of reported outbreaks include Staphylococcus aureus, herpes simplex virus, Streptococcus pyogenes, and several fungi. Although once thought of solely as a nosocomial pathogen, methicillin-resistant Staphylococcus aureus has been identified as an emerging problem in the community, particularly in the athletic population. Despite a recent increase in reported outbreaks of community acquired methicillin-resistant Staphylococcus aureus soft tissue infection in athletic teams, many sports medicine physicians are unfamiliar with the epidemiology of this pathogen. It is spread via person-to-person contact and is harbored within the anterior nares and on the skin of carriers. Outbreaks of community-acquired methicillin-resistant Staphylococcus aureus soft tissue infection are not treated by traditional beta-lactam antibiotics, and they can be difficult to eradicate. Such infections have been associated with significant morbidity, with up to 70% of involved team members requiring hospitalization and intravenous antibiotics. A thorough understanding of community-acquired methicillin-resistant Staphylococcus aureus is essential for the sports medicine physician to properly identify, treat, and control infectious outbreaks. PMID- 16314669 TI - Irreducible posterolateral knee dislocation associated with interposition of the vastus medialis. PMID- 16314670 TI - Reconstituted high density lipoprotein enriched with the polyene antibiotic amphotericin B. AB - The polyene antibiotic amphotericin B (AMB) is an effective antifungal agent whose therapeutic potential is limited by poor aqueous solubility and toxicity toward host tissues. Addition of apolipoprotein A-I to a multilamellar phospholipid vesicle dispersion containing 20% (w/w) AMB induces the formation of reconstituted high density lipoprotein (rHDL), with solubilization of the antibiotic. Density gradient ultracentrifugation resulted in flotation of the complexes to a density of 1.16 g/ml, and negative stain electron microscopy revealed a population of disk-shaped particles. Native gradient polyacrylamide gel electrophoresis indicated a particle diameter of approximately 8.5 nm. Absorbance spectroscopy provided evidence for AMB integration into the lipid milieu. AMB-rHDLs were potent inhibitors of Saccharomyces cerevisiae growth, yielding 90% growth inhibition at <1 microg/ml yeast culture. In studies with pathogenic fungal species, similar growth inhibition characteristics were observed. Compared with AMB-deoxycholate micelles, AMB-rHDL displayed greatly attenuated red blood cell hemolytic activity and decreased toxicity toward cultured hepatoma cells. In in vivo studies in immunocompetent mice, AMB-rHDLs were nontoxic at 10 mg/kg, and they showed efficacy in a mouse model of candidiasis at concentrations as low as 0.25 mg/kg. These results indicate that AMB-rHDLs constitute a novel formulation that effectively solubilizes the antibiotic and elicits strong in vitro and in vivo antifungal activity with no observed toxicity at therapeutic doses. PMID- 16314671 TI - Missed opportunity for osteopathic medical education. PMID- 16314672 TI - The "Big DO". PMID- 16314673 TI - "Opportunities" for improvement. PMID- 16314674 TI - Jumping through hoops for osteopathic internships. PMID- 16314675 TI - Graduate medical education: accuracy of AOA's annual data-reporting mechanisms questioned. PMID- 16314676 TI - Surprised by surgery statistics. PMID- 16314677 TI - Increased lymphatic flow in the thoracic duct during manipulative intervention. AB - The thoracic pump and the abdominal pump are osteopathic manipulative (OM) lymphatic pump techniques frequently used by osteopathic physicians to treat patients with infections (eg, pneumonia, otitis media). Although there is a widely accepted belief among the osteopathic medical profession that increasing lymphatic flow is beneficial, no measurements of lymph flow during osteopathic manipulative treatment have been reported. The authors surgically instrumented five mongrel dogs to record lymphatic flow in the thoracic duct (TDF) and cardiac variables during three intervention protocols. After recovery from surgery, canine subjects were placed in a standing-support sling, and TDF, cardiac output, mean aortic blood pressure, and heart rate were recorded during two randomized 30 second sessions of manipulative intervention using the osteopathic thoracic pump and abdominal pump techniques on two successive days. Lymph flow in the thoracic duct increased from 1.57+/-0.20 mL x min(-1) to a peak TDF of 4.80+/-1.73 mL x min(-1) during abdominal pump, and from 1.20+/-0.41 mL x min(-1) to 3.45+/-1.61 mL x min(-1) during thoracic pump. Lymph flow in the thoracic duct and cardiac variables were also recorded for canine subjects during physical activity (ie, treadmill exercise at 3 miles per hour at 0% incline). During physical activity, TDF increased from 1.47+/-0.33 mL x min(-1) to 5.81+/-1.30 mL x min(-1). Although cardiac variables did not change significantly during manipulative intervention with lymphatic pump techniques, cardiac output and heart rate did increase during physical activity. The authors conclude that physical activity and manipulative intervention using thoracic pump and abdominal pump techniques produced net increases in TDF (P<.05). PMID- 16314678 TI - Manual techniques addressing the lymphatic system: origins and development. PMID- 16314679 TI - Interobserver reliability of osteopathic palpatory diagnostic tests of the lumbar spine: improvements from consensus training. AB - CONTEXT: Establishing reliable palpatory tests continues to be a critical, yet elusive, step in osteopathic medical research and evidence-based clinical practice. OBJECTIVE: The authors investigated the interobserver reliability of common osteopathic palpatory tests used to evaluate the lumbar spine. DESIGN AND METHODS: Subjects (N=119) were recruited from the faculty, staff, and students of Kirksville (MO) College of Osteopathic Medicine (KCOM) of A.T. Still University of Health Sciences. Three osteopathic medical examiners residency trained in neuromusculoskeletal medicine initially evaluated lumbar segments on subjects from one subgroup (n=42) in a blinded assessment. The examiners performed palpatory tests of tenderness and tissue texture changes, as well as--in three planes--vertebral positional asymmetry and motion asymmetry. Kappa statistics (kappa) were used to evaluate interobserver reliability. Following a period of consensus training, subjects from another subgroup (n=77) were evaluated in a blinded assessment for those palpatory tests that seemed most likely to produce reliable findings. The interobserver reliability was then re-evaluated. RESULTS: During the initial evaluation of interobserver reliability, kappa ranged from 0.02 to 0.34, within the poor-to-fair reliability range. Following consensus training, reliability improved, rising into the moderate range for tissue texture changes (kappa=0.45) and into the substantial range for tenderness assessments (kappa=0.68). Reliability for positional asymmetry in the transverse plane (kappa=0.34) and rotational motion asymmetry (kappa=0.20) were improved but remained in the fair range. CONCLUSION: The authors concluded that consensus training improved the interobserver reliability of common osteopathic palpatory tests of the lumbar spine. In two of the four tests that were studied--tissue texture and tenderness--acceptable kappa values for clinical tests were achieved after consensus training. PMID- 16314680 TI - Hemodynamic effects of osteopathic manipulative treatment immediately after coronary artery bypass graft surgery. AB - CONTEXT: Coronary artery bypass graft (CABG) surgery is a common procedure for patients with coronary artery disease. The physiologic effects of postoperative osteopathic manipulative treatment (OMT) following CABG have not been documented previously. OBJECTIVE: To determine the effects of OMT on cardiac hemodynamics post-CABG surgery. DESIGN: Pilot prospective clinical study (N=29). SETTING AND PATIENTS: Treatment subjects (n=10) undergoing CABG surgery were recruited for postoperative OMT. The primary assessment compared, pre-OMT versus post-OMT, measurements of thoracic impedance, mixed venous oxygen saturation (SvO2), and cardiac index. Records of control subjects (n=19) who underwent CABG surgery--but who did not receive OMT--were assessed for SvO2 and cardiac index at 1 hour and 2 hours postsurgery. INTERVENTION: Immediately following CABG surgery (< or = 2 h), OMT was provided to subjects to alleviate anatomic dysfunction of the rib cage caused by median sternotomy and to improve respiratory function. This adjunctive treatment occurred while subjects were completely anesthetized. RESULTS: A post OMT increase in thoracic impedance (P < or = .02) in OMT subjects demonstrated that central blood volume was reduced after OMT, suggesting an improved peripheral circulation. Mixed venous oxygen saturation also increased (P < or = .005) after OMT. These increases were accompanied by an improvement in cardiac index (P < or = .01). Comparisons of postoperative measurements in OMT subjects versus those in control subjects revealed statistically significant differences for SvO2 (P < or = .005) and cardiac index (P < or = .02) between the two groups. CONCLUSION: The observed changes in cardiac function and perfusion indicated that OMT had a beneficial effect on the recovery of patients after CABG surgery. The authors conclude that OMT has immediate, beneficial hemodynamic effects after CABG surgery when administered while the patient is sedated and pharmacologically paralyzed. PMID- 16314681 TI - Evolutionary relationships among basal fungi (Chytridiomycota and Zygomycota): Insights from molecular phylogenetics. AB - Evolutionary relationships of the two basal fungal phyla Chytridiomycota and Zygomycota are reviewed in light of recent molecular phylogenetic investigation based on rDNA (nSSU, nLSU rDNA), entire mitochondrial genomes, and nuclear protein coding gene sequences (e.g., EF-1alpha, RPB1). Accumulated molecular evidence strongly suggests that the two basal fungal phyla are not monophyletic. For example, the chytridiomycete order Blastocladiales appears to be closely related to the zygomycete order Entomophthorales. Within the Zygomycota, a monophyletic clade, consisting of the Dimargaritales, Harpellales, and Kickxellales, which is characterized by a shared unique septal ultrastructure, was identified. Moreover, evidence for the exclusion of zygomycete orders Amoebidiales and Eccrinales from the Fungi, and their placement at the Animal Fungi boundary has been clearly documented. Microsporidia, a group of amitochondriate organisms currently under intensive study, is not supported as derived within the Fungi, but a fungal affinity cannot be ruled out. Taking these molecular phylogenetic studies into account, we proposed a hypothetical evolutionary framework of basal fungi. PMID- 16314682 TI - Sporobolomyces diospyroris sp. nov., Sporobolomyces lophatheri sp. nov. and Sporobolomyces pyrrosiae sp. nov., three new species of ballistoconidium-forming yeasts in the Agaricostilbum lineage isolated from plants in Taiwan. AB - Three strains of xylose-lacking and ubiquinone-10-having ballistoconidium-forming yeasts isolated from plant leaves collected in Taiwan were found to represent respective new species. In phylogenetic trees constructed based on the nucleotide sequences of 18S rDNA and D1/D2 domain of 26S rDNA, they were located in the Agaricostilbum lineage (Agaricostilbum/Bensingtonia cluster). Since the taxonomic properties of these species coincide with those of the genus Sporobolomyces, they are described as Sporobolomyces diospyroris sp. nov., Sporobolomyces lophatheri sp. nov. and Sporobolomyces pyrrosiae sp. nov., respectively. PMID- 16314683 TI - Reclassification of Methylobacterium chloromethanicum and Methylobacterium dichloromethanicum as later subjective synonyms of Methylobacterium extorquens and of Methylobacterium lusitanum as a later subjective synonym of Methylobacterium rhodesianum. AB - Phylogenetic analysis based on 16S rDNA sequences was performed on all type strains of the 14 validly described Methylobacterium species to ascertain the genealogic relationships among these species. The results showed that type strains of Methylobacterium were divided into two monophyletic groups whose members were distinct species with sequence similarity values greater than 97.0% between any two of the members in the same group. Only M. organophilum JCM 2833(T) and ATCC 27886(T) were not divided into those two groups. In particular, strains of M. dichloromethanicum and M. chloromethanicum exhibited extremely high similarity values (99.9 and 100%, respectively) with the type strain of M. extorquens. To clarify the relationships among Methylobacterium species in more detail, phylogenetic analysis based on the 5' end hyper-variable region of 16S rDNA (HV region), ribotyping analysis, fatty acid analysis, G+C content analysis and DNA-DNA hybridization experiments was performed on 58 strains of Methylobacterium species. Results of the ribotyping analysis and the phylogenetic analysis based on HV region sequences indicated that many Methylobacterium strains, including M. 'organophilum' DSM 760(T), have been erroneously identified. The DNA G+C content of Methylobacterium strains were between 68.1 and 71.3%. Results of whole-cell fatty-acid profiles showed that all strains contained 18 : 1omega7c as the primary fatty acid component (82.8-90.1%), with 16 : 0 and 18 : 0 as minor components. M. dichloromethanicum DSM 6343(T), M. chloromethanicum NCIMB 13688(T), and M. extorquens IAM 12631(T) exhibited high DNA-DNA relatedness values between each other (69-80%). M. lusitanum NCIMB 13779(T) also showed a close relationship with M. rhodesianum DSM 5687(T) at DNA DNA relatedness levels of 89-92%. According to these results, many Methylobacterium strains should be reclassified, with M. dichloromethanicum and M. chloromethanicum regarded as a synonym of M. extorquens, and M. lusitanum a synonym for M. rhodesianum. PMID- 16314684 TI - Neoasaia chiangmaiensis gen. nov., sp. nov., a novel osmotolerant acetic acid bacterium in the alpha-Proteobacteria. AB - An acetic acid bacterium, designated as isolate AC28(T), was isolated from a flower of red ginger (khing daeng in Thai; Alpinia purpurata) collected in Chiang Mai, Thailand, at pH 3.5 by use of a glucose/ethanol/acetic acid (0.3%, w/v) medium. A phylogenetic tree based on 16S rRNA gene sequences for 1,376 bases showed that isolate AC28(T) constituted a cluster along with the type strain of Kozakia baliensis. However, the isolate formed an independent cluster in a phylogenetic tree based on 16S-23S rDNA internal transcribed spacer (ITS) region sequences for 586 bases. Pair-wise sequence similarities of the isolate in 16S rRNA gene sequences for 1,457 bases were 93.0-88.3% to the type strains of Asaia, Kozakia, Swaminathania, Acetobacter, Gluconobacter, Gluconacetobacter, Acidomonas, and Saccharibacter species. Restriction analysis of 16S-23S rDNA ITS regions discriminated isolate AC28(T) from the type strains of Asaia and Kozakia species. Cells were non-motile. Colonies were pink, shiny, and smooth. The isolate produced acetic acid from ethanol. Oxidation of acetate and lactate was negative. The isolate grew on glutamate agar and mannitol agar. Growth was positive on 30% D-glucose (w/v) and in the presence of 0.35% acetic acid (w/v), but not in the presence of 1.0% KNO(3) (w/v). Ammoniac nitrogen was hardly assimilated on a glucose medium or a mannitol medium. Production of dihydroxyacetone from glycerol was weakly positive. The isolate did not produce a levan-like polysaccharide on a sucrose medium. Major isoprenoid quinone was Q-10. DNA base composition was 63.1 mol% G+C. On the basis of the results obtained, Neoasaia gen. nov. was proposed with Neoasaia chiangmaiensis sp. nov. The type strain was isolate AC28(T) (=BCC 15763(T) =NBRC 101099(T)). PMID- 16314685 TI - Isolation and characterization of lactic acid bacteria from soils in vineyards. PMID- 16314686 TI - ITS sequence and electrophoretic karyotype comparisons of Candida ethanolica with Pichia deserticola and Candida odintsovae with Pichia rabaulensis. PMID- 16314687 TI - SKG6, a suppressor gene of synthetic lethality of kex2Delta gas1Delta mutations, encodes a novel membrane protein showing polarized intracellular localization. PMID- 16314688 TI - Antinociceptive effect of shakuyakukanzoto, a Kampo medicine, in diabetic mice. AB - In this study, the antinociceptive effect of shakuyakukanzoto was investigated using streptozotocin-induced diabetic mice to certify its analgesic effect on diabetic patients. Shakuyakukanzoto (0.5 and 1.0 g/kg, p.o.) significantly increased the nociceptive threshold in diabetic mice. The antinociceptive activity of shakuyakukanzoto in diabetic mice was not antagonized by beta funaltrexamine, naltrindole, or nor-binaltorphimine. The increased antinociceptive activity of (1.0 g/kg, p.o.) in diabetic mice was abolished by yohimbine (15 microg, i.t.), but not by NAN-190 (1 microg, i.t.), methysergide (15 microg, i.t.), or MDL-72222 (15 microg, i.t.). In shakuyakukanzoto diabetic mice treated with 6-hydroxydopamine (20 microg, i.t.) chemically lesioned noradrenergic pathways, shakuyakukanzoto (1.0 g/kg, p.o.) failed to exhibit an antinociceptive effect. Furthermore, the antinociceptive activity induced by norepinephrine (0.06 - 2 microg, i.t.) was markedly more potent in diabetic mice than in non-diabetic mice at the same dose. These results suggest that the antinociceptive effect of shakuyakukanzoto in diabetic mice is not mediated by the opioid systems and that this effect appears via selective activation of the spinal descending inhibitory alpha2-adrenergic systems without activating the serotonergic systems. The spinal alpha2-adrenoceptor-mediated analgesic mechanism was enhanced in diabetic mice, suggesting that shakuyakukanzoto exhibits its effect by activating the descending noradrenergic neurons. PMID- 16314689 TI - Sclerosing effect of OC-108, a novel agent for hemorrhoids, is associated with granulomatous inflammation induced by aluminum. AB - OC-108 is a novel sclerosing agent for hemorrhoids, containing aluminum potassium sulfate (alum) and tannic acid as its main ingredients. In clinical studies, OC 108 injection therapy for severe internal hemorrhoids proved to be highly effective, not only on bleeding but also for prolapse, and the effects were comparable to hemorrhoidectomy. The aim of this study was to elucidate the mode of action by administrating the agent s.c. to mice and rats. In response to OC 108 injection, inflammation with necrosis developed at an early stage followed by granuloma formation with fibrosis at the injection site. Necrotic debris with aluminum was observed in the granuloma for a long period. Alum, as well as OC 108, induced vascular permeability, leukocyte infiltration, and granuloma formation; however, tannic acid did not. On the other hand, tannic acid inhibited leukocyte infiltration induced by alum but did not inhibit granuloma formation. These results indicate that OC-108 causes sclerosis and retraction of hemorrhoids through fibrosis associated with granulomatous chronic inflammation induced by the main active ingredient alum and that the adjunct ingredient tannic acid reduces excessive acute inflammation induced by alum. PMID- 16314690 TI - FK614, a novel peroxisome proliferator-activated receptor gamma modulator, induces differential transactivation through a unique ligand-specific interaction with transcriptional coactivators. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand dependent transcriptional factor implicated in regulating adipogenesis, glucose homeostasis, and in mediating the action of the insulin sensitizing anti-diabetic thiazolidinedione (TZD) compounds. [3-(2,4-Dichlorobenzyl)-2-methyl-N (pentylsulfonyl)-3-H-benzimidazole-5-carboxamide] (FK614) is a structurally novel PPARgamma agonist that demonstrates potent anti-diabetic activity in vivo. Herein, we describe that FK614 is a selective PPARgamma ligand with specific transactivation properties that are dependent upon the context of coactivators. FK614 dissociates the corepressors NCoR (nuclear receptor corepressor) and SMRT (silencing mediator of retinoid and thyroid hormone receptors) from PPARgamma as effectively as rosiglitazone and pioglitazone, but can also differentially induce a ligand specific interaction of PPARgamma with coactivators. The amount of CBP (CREB-binding protein) and SRC-1 (steroid receptor coactivator-1) recruited by FK614 was less than that induced by rosiglitazone and pioglitazone, but FK614 caused similar PGC-1alpha (PPARgamma coactivator-1alpha) recruitment as these compounds. As a consequence of these ligand-specific differences in the strength of ligand-type specific interactions of PPARgamma and coactivators, FK614 functions as a partial or full agonist for transcriptional activation depending upon the amount of specific coactivators in cells following overexpression. In conclusion, FK614 is a novel, non-TZD type, and selective PPARgamma modulator whose pharmacological properties are distinct from rosiglitazone and pioglitazone. PMID- 16314691 TI - Milk whey culture with Propionibacterium freudenreichii ET-3 is effective on the colitis induced by 2,4,6-trinitrobenzene sulfonic acid in rats. AB - This study aimed to evaluate whether milk whey culture with Propinibacterium freudenreichii ET-3 (milk whey culture), which has been reported to have Bifidogenic activity, is effective on the colitis induced by 2,4,6 trinitrobenzene sulfonic acid (TNBS) in rats. For the induction of colitis, the colon was clamped and 0.1 M TNBS in 35% ethanol was injected into the luminal side of the clamped portion under pentobarbital anesthesia. From the next day of colitis induction, milk whey culture was administered orally at doses of 1 and 3 g/kg, twice a day for 9 days. On the 10th day, rats were sacrificed and ulcer size was measured. Milk whey culture significantly accelerated the healing of the colitis in a dose-dependent manner, but culture medium did not. To clarify the active substance, the effects of propionic acid and acetic acid contained in milk whey culture was tested. Sodium propionate significantly accelerated the healing of TNBS-induced colitis, but sodium acetate did not. The above results show that milk whey culture may become a useful prebiotic for the therapy of inflammatory bowel disease and that propionic acid may be one of the active substances contained in milk whey culture. PMID- 16314692 TI - Effect of antiepileptic drugs on the urinary excretion of porphyrins in non porphyric subjects. AB - The action of some anticonvulsant drugs as the causal agents of attacks of acute porphyria has been widely documented in the literature. However, little attention has been paid to the effect of these drugs on the urinary excretion of porphyrins in non-porphyric subjects. In a sample of 82 epileptic patients treated with phenobarbital (n = 54), phenytoin (n = 64), carbamazepine (n = 33), and valproate (n = 8), the daily doses were expressed according to a drug score that would reflect the capacity of these drugs as enzymatic inducers when administered in polytherapy. A significantly increased urinary excretion of D-glucaric acid (DGA) and porphyrins was found in this group of patients (P<0.001), with coproporphyrin being the major fraction in all cases (>60%). Urinary DGA had a highly significant correlation with the drug score (r = 0.783, P<0.001); however, no significant correlations were found between the urinary porphyrins and DGA (r = 0.005) or the drug score (r = 0.053). Neither was any significant relationship found between the urinary porphyrins and the serum activity of 5'-nucleotidase (r = 0.066) or the presence of a cholestasis objectivized through the presence of the isoform of gamma-glutamyltransferase with beta-globulins electrophoretic mobility. However, in a group of 10 patients a significant correlation was found between the urinary excretion of porphyrins and beta-N-acetylhexosaminidase (r = 0.790, P<0.01). Therefore, it does not appear that the liver enzyme induction, or even a subclinical cholestasis, produced by the antiepileptic drugs administered to these patients may serve to explain the increase in the urinary excretion of porphyrins. A possible renal origin is proposed for the increase of urinary porphyrins in these cases. PMID- 16314694 TI - Future directions in body magnetic resonance imaging. AB - Technologic innovations in instrumentation and contrast agents naturally lead to new clinical and research applications in body MRI. Although long-range predictions of innovation are an uncertain process, short-term trends in development are more readily discernable. This review will provide examples of recent developments in magnetic resonance spectroscopic imaging, contrast agent development and molecular imaging, instrumentation, post-processing, and screening in an attempt to describe areas of active research. PMID- 16314695 TI - Whole-body magnetic resonance imaging. AB - Whole-body magnetic resonance imaging is a fast and accurate modality for the detection of disease throughout the entire body. Technical improvements including the availability of different high image quality MR sequences, the remote movement of the imaging table, and the use of specialized surface coils have rendered whole-body screening with MRI a feasible method. In this article we describe underlying techniques and report on first clinical experiences of whole body magnetic resonance imaging as a staging and screening method. Furthermore, advantages and limitations compared with whole-body imaging based on computed tomography are discussed. PMID- 16314696 TI - Magnetic resonance angiography of abdominal and lower extremity vasculature. AB - Magnetic resonance angiography (MRA) has evolved over the past years from an experimental imaging modality to a technique that is now widely applied in clinical practice. This article reviews the fundamentals of the different magnetic resonance angiographic techniques and how they can be applied for abdominal and peripheral arterial imaging. Currently, contrast-enhanced magnetic resonance angiography (CE-MRA), whereby a luminogram is obtained during initial arterial passage of contrast material, is the most widely used technique. With current hardware and software, high-spatial resolution images of the abdominal aorta and proximal visceral branches can be obtained that are equivalent to intra arterial digital subtraction angiography (IA-DSA). High-quality imaging of the renal arteries demands isotropic voxels and reformations orthogonal to the vessel axis for evaluation. Contrast-enhanced magnetic resonance angiography of the peripheral vascular tree is now a highly accurate technique and has replaced diagnostic intra-arterial digital subtraction angiography and duplex ultrasonography in many hospitals. PMID- 16314697 TI - Magnetic resonance imaging and expression of vascular endothelial growth factor in hepatocellular nodules in cirrhosis and hepatocellular carcinomas. AB - We summarized and discussed our previous research results on correlation between magnetic resonance (MR) imaging findings and vascular endothelial growth factor (VEGF) expression in benign or borderline hepatocellular nodules in cirrhosis, hepatocellular carcinomas (HCCs), and in the surrounding liver. Magnetic resonance images were retrospectively correlated quantitatively and qualitatively with VEGF expression in hepatic nodules and in the surrounding liver. By immunohistochemistry, hepatic nodules with moderate to strong immunoreactivity for VEGF showed higher T1 signal intensity, and those with intense immunoreactivity for VEGF showed higher T2 signal intensity. By Western blotting, HCC-to-liver contrast-to-noise ratio correlated with VEGF indices (VEGFs) of hepatocellular carcinomas inversely on opposed-phase T1-weighted, directly on T2 weighted, and marginally and inversely on gadolinium-enhanced hepatic arterial phase images. On T2-weighted images, standard-deviation ratio of hepatocellular carcinomas correlated directly with VEGFs of hepatocellular carcinomas. Heterogeneities of hepatocellular carcinomas on MR images correlated directly with VEGFs of HCCs on opposed-phase T1-weighted, T2-weighted, hepatic arterial phase, and equilibrium-phase images. Our results may reflect that MR signal intensity, hepatic arterial vascularity, and heterogeneity of hepatocellular nodules on MR images are closely related to the intensity of VEGF expression as up-regulated by hyper- or hypoxia in the nodules. Gadolinium-enhanced MR imaging may be useful to monitor ischemic state of hepatocelluar nodules. Although real impacts of our results on radiologic practice have been still debatable, we believe that our results may help future radiologic practice in conjunction with biomolecular or genetic treatments for hepatocellular carcinomas. PMID- 16314698 TI - Magnetic resonance imaging of the gastrointestinal tract. AB - Comprehensive bowel examination results from the combined use of T2-weighted single-shot and breath hold T1-weighted gradient echo, minus/plus fat suppression, and gadolinium-enhanced 3D gradient echo (3D VIBE, T1 FAME, 3D THRIVE). Gadolinium-enhanced imaging should be performed dynamically, but the venous 60- to 90-second delayed phase images with fat suppression are generally the most valuable. Removal of fat signal for detection of enhancing normal and abnormal structures is critical. Newly available True-FISP (FIESTA, BFFE) sequences obtained in the 2D form can be very helpful in delineation of bowel wall pathology and overall bowel anatomy, particularly when combined with a water based intraluminal distending agent. Advantages include rapid acquisition, high signal-to-noise, and motion insensitivity. Generalized protocol for comprehensive evaluation of the entire abdomen and pelvis can be used for the following bowel indications: type and severity of inflammatory bowel disease (IBD); identifying enteric abscesses and fistulae; preoperative staging of malignant neoplasms, including rectal carcinoma; differentiating postoperative and radiation therapy changes from recurrent carcinoma; follow-up evaluation of metastases response to localized ablative or systemic chemotherapy. For improved visualization of bowel wall in dedicated examinations, bowel distension should be achieved using either orally or rectally delivered contrast agents to produce either bright or dark lumen. We have found 2D True-FISP without fat suppression superior to 3D True FISP and to single-shot echo-train sequences to provide a T2-weighted image of bowel morphology. Strengths include: performed without fat suppression results in the very dark bowel wall being sandwiched between intermediate high signal fat adjacent to bowel serosa, and very high lumen signal from water-distending agent; 2D True-FISP provides motion insensitivity that is lost if 3D is used; True-FISP produces better edge sharpness than single-shot echo-train, higher contrast, and resists flow void artifacts commonly seen with single-shot echo-train imaging combined with a water distending agent. Drawbacks of this technique include: artifacts related to extreme sensitivity to field inhomogeneity, including air soft tissue interfaces at the patient skin surface, and from retained bowel gas; retained bowel gas is dark against dark bowel wall, impairing bowel wall assessment; and True-FISP does not provide sensitivity for edema, which is superior on single-shot echo-train imaging. Small/large bowel indications for MRI include: inflammatory bowel disease, infectious disease including abscess evaluation or for appendicitis, inflammatory conditions including ischemia, and partial obstruction, malnutrition, and neoplasm search. PMID- 16314699 TI - Magnetic resonance imaging features of focal liver lesions after intervention. AB - The aim of this article is to provide an overview of MRI features of primary and secondary liver lesions after intervention. Emphasis is given to T2- and T1 weighted pre and post intravenous gadolinium chelates appearance of successfully treated lesions and, from residual/recurrent tumors after therapy. In addition, complications after procedures are briefly cited. Liver resection, radiation therapy, systemic chemotherapy, transcatheter arterial chemoembolization, ablative therapies and liver transplantation are the methods discussed in this review. Among these methods, special attention is given to MRI findings after ablative therapies since radiofrequency ablation is commonly performed in our institution. PMID- 16314700 TI - Contrast agents in abdominal imaging: current and future directions. AB - Magnetic resonance imaging is an established imaging method for the evaluation of the abdomen. Accurate assessment of the liver, spleen, pancreas, bile ducts, vascular structures, and retroperitoneal organs (eg, the kidneys, the collecting system, and the adrenals) are possible on MR imaging. The intravenous administration of MR contrast agents can frequently improve the examination and provide more specific diagnoses. The advent of more specific, "hepatobiliary" contrast agents has further improved the differential diagnostic process, particularly for MR imaging of the liver. The availability of orally administered MR contrast agents has further extended the range of abdominal applications, making MR imaging of the small bowel and the colon established imaging procedures. PMID- 16314701 TI - Spitz nevus is relatively frequent in adults: a clinico-pathologic study of 247 cases related to patient's age. AB - Spitz nevus is a clinico-pathologic entity that can cause diagnostic concern, particularly in adults. Many studies have been performed to establish reliable histologic criteria, in the attempt to differentiate this lesion from melanoma. A series of 247 Spitz nevi, 6 of which were formerly classified as melanomas, were reviewed for clinical and histopathological parameters. Patients older than 20 comprised 66% of cases, with a predominance of women. The lower extremity was more affected in females of any age, whereas the trunk was more frequently involved in men over 40. Histopathologic examination showed the following differences among Spitz nevi related to age: acanthosis, parakeratosis, pagetoid infiltration, and Kamino bodies were more frequent in young people, whereas multinucleated melanocytes were more frequent in adults. The latter also had lesions that were less pigmented, with less maturation and more desmoplasia. At a mean follow-up of 94 months (range 52-172), recurrence at the site of biopsy or metastases were absent. In our study, a greater proportion of Spitz nevi occurred in adults than in previous series. Moreover, the relative incidence of Spitz nevus compared with melanoma in our population was higher than in other studies. Histopathologic criteria elaborated to diagnose Spitz nevus, applied to our cases, appeared reliable, allowing a correct diagnosis, even in adults. PMID- 16314702 TI - Keratin 16 expression in epidermal melanocytes of normal human skin. AB - Although the prevailing dogma states that keratin filaments are the hallmark of keratinocytes and other epithelial cells, recent publications suggest that they may be expressed by a variety of normal and malignant cells of different embryonic origin. Keratin expression has been reported in fibroblasts and endothelial cells as well as in various sarcomas. Also, some human melanomas express keratins in addition to the traditional diagnostic markers of differentiation, such as S-100 and melanocyte-specific antigens. Many studies have shown that cultured cells obtained from various melanomas express keratin. Most recently, keratin expression has also been shown in cultured melanocytes of normal skin. We now report that normal human melanocytes in vivo express keratin 16 (K16) but not keratins 1, 5, 8, 10, 14, or even keratin 6, the type II partner that is normally expressed with K16 in keratinocytes. Similarly, melanocytes in vitro express K16 but not K6. Keratin 16 expression in vivo was present in basal melanocytes in specimens derived from donors (0-77 years) and from different anatomic locations, suggesting that keratin 16 is constitutively expressed by all melanocytes. It appears that keratin expression may be more prevalent than previously assumed, and that these cytoskeletal filaments may play important roles in tissues and cells other than epithelia. PMID- 16314703 TI - Paratumoral epidermal hyperplasia: a novel prognostic factor in thick primary melanoma of the skin? AB - There is a lack of histopathological factors to sub-stratify prognosis in pT3/4 melanoma primaries. In the presented pilot study, the prognostic significance of different clinical and histopathological parameters was studied in thick primary melanoma taking paratumoral epidermal hyperplasia (PTEH) into consideration. Of 1632 melanoma patients in the melanoma register of the Martin Luther University Halle-Wittenberg in the years 1980 to 1987, 16 cases with tumor thickness (TT) of the primary > or = 3 mm, documented metastasis-free follow-up of 10+ years after primary therapy and available histologic sections were compared with an adequate recurrence control group (n = 62) by PTEH and standard prognostic parameters. PTEH was demonstrable in 15 of 16 patients of the metastasis-free group (PTEH penetration depth 1.42 +/- 0.82 mm/mean +/- SD) and 27 of 62 of controls (0.29 +/ 0.46 mm), P < or = 0.001. Of the standard prognostic parameters, TT, sex, location, and lack of nevus association also correlated with metastasis. In multivariate analysis, PTEH > or = 1 mm was the single independent parameter with the highest (negative) association to recurrence (odds ratio 52.3). Occurrence of PTEH might predict a more moderate course of disease in thick melanoma. Thus, it might become an easily determinable and effective tool to sub-stratify prognosis in thick primary melanoma of the skin. Further studies are necessary to prove these findings. PMID- 16314704 TI - Alterations of basement membrane zone and cutaneous microvasculature in morphea and extragenital lichen sclerosus. AB - The aim of this study was to compare alterations of the basement membrane zone (BMZ) and to visualize changes within the skin vascular network in morphea and extragenital lichen sclerosus with the use of laser scanning confocal microscopy. This work was performed in eight plaques of morphea (three active and five inactive) and eight of lichen sclerosus (three of short duration and five long lasting). Biopsy specimens from six healthy individuals served as controls. The biopsies were cut into 40-microm-thick sections, labeled with antibodies against beta4-intergin (a lamina lucida marker), collagen IV, and the N-terminal end of collagen VII (lamina densa markers) and C-terminal end of collagen VII (a sublamina densa marker) and studied using laser scanning confocal microscopy. Three-dimensional reconstruction of various regions of the BMZ showed a decreased number and size of the dermal papillae both in morphea and lichen sclerosus compared with normal skin. In morphea, the continuity of the BMZ at the level of lamina lucida, lamina densa, and sublamina densa was preserved whereas in LS numerous invaginations and holes were present in the BMZ at the level of the lamina lucida and lamina densa. Thus the alterations of the BMZ in morphea differ from those in lichen sclerosus. Three-dimensional reconstruction of the skin vascular network showed increased angiogenesis only in the early inflammatory stage of morphea, whereas in inactive morphea and lichen sclerosus various numbers of enlarged vessels were visible. The changes in the vascular network in morphea appear to be related to the activity of the disease. PMID- 16314705 TI - Glomus coccygeum: report of a case and review of the literature. AB - The glomus coccygeum is a vestigial structure related to the canals of Sucquet Hoyer, an arteriovenous anastomosis surrounded by glomus cells derived from modified smooth muscle and involved in thermoregulation. It is an incidental finding in specimens from the sacral area and may represent a diagnostic challenge to the unaware observer. We present a case of a glomus coccygeum, presenting as a 1.5-mm structure adjacent to a typical pilonidal cyst excised from a 7-month-old boy, which was the subject of a second opinion consultation. The lesion showed small to medium sized clusters of predominantly epithelioid cells with moderate amounts of clear to eosinophilic cytoplasm, intercellular borders, and plump, round nuclei with fine chromatin. These cells were closely associated with small vascular channels and nerves. Immunohistochemistry revealed that the epithelioid cells expressed vimentin, muscle-specific actin, neuron specific enolase, and S-100 protein, were weakly positive for smooth muscle actin, and negative for desmin, synaptophysin, and chromogranin. The endothelial cells of the vascular channels were antibody CD31 positive. Recognition of the histologic features of glomus coccygeum is important to avoid confusion with glomus tumor and neural or smooth muscle neoplasms in the sacral area. PMID- 16314706 TI - Porocarcinoma arising in pigmented hidroacanthoma simplex. AB - Hidroacanthoma simplex (HAS) is a rare benign tumor that is also known as intraepidermal poroma. While there have been a few reports of HAS with malignant transformation (porocarcinoma), we report an unusual case of porocarcinoma, arising in a pigmented HAS, the latter also showing secondary amyloid deposits. An 80-year-old Japanese man presented with a cutaneous tumor on his left buttock, which had first been noticed in his childhood. The tumor consisted of flat pigmented plaque and a depigmented papule with erosion. Histologic analysis revealed many pigmented and well-defined nests within the epidermis of the flat pigmented portion. The nests were composed of cuboidal to oval and occasionally elongated, bland, basaloid cells with numerous melanin granules. In addition, there were infrequently ductal structures and small clusters of sebocytes, and abundant amyloid deposits in the upper dermis. These findings were consistent with pigmented HAS with amyloid deposition. In the depigmented portion, markedly atypical cells with occasional ductal structures and intracytoplasmic lumina extended throughout the entire thickness of the epidermis, with minimal invasion of the dermis. We considered this portion of the tumor to be a porocarcinoma. Since the two portions of the tumor were continuous, we made a final diagnosis of porocarcinoma arising in pre-existing pigmented HAS with amyloid deposition. PMID- 16314707 TI - Cutaneous vasculitis update: diagnostic criteria, classification, epidemiology, etiology, pathogenesis, evaluation and prognosis. AB - Vasculitis, inflammation of the vessel wall, can result in mural destruction with hemorrhage, aneurysm formation, and infarction, or intimal-medial hyperplasia and subsequent stenosis leading to tissue ischemia. The skin, in part due to its large vascular bed, exposure to cold temperatures, and frequent presence of stasis, is involved in many distinct as well as un-named vasculitic syndromes that vary from localized and self-limited to generalized and life-threatening with multi-organ disease. To exclude mimics of vasculitis, diagnosis of cutaneous vasculitis requires biopsy confirmation where its acute signs (fibrinoid necrosis), chronic signs (endarteritis obliterans), or past signs (acellular scar of healed arteritis) must be recognized and presence of extravascular findings such as patterned fibrosis or collagenolytic granulomas noted. Although vasculitis can be classified by etiology, many cases have no identifiable cause, and a single etiologic agent can elicit several distinct clinicopathologic expressions of vasculitis. Therefore, the classification of cutaneous vasculitis is best approached morphologically by determining vessel size and principal inflammatory response. These histologic patterns roughly correlate with pathogenic mechanisms that, when coupled with direct immunofluorescent examination, anti-neutrophil cytoplasmic antibody (ANCA) status, and findings from work-up for systemic disease, allow for specific diagnosis, and ultimately, more effective therapy. Herein, we review cutaneous vasculitis focusing on diagnostic criteria, classification, epidemiology, etiology, pathogenesis, and evaluation of the cutaneous vasculitis patient. PMID- 16314708 TI - History: Frederic Woringer (1903-1964) and Woringer-Kolopp disease. AB - The two most famous dermatologic eponyms born in Strasbourg are Pautrier microabscess and Woringer-Kolopp disease. Frederic Woringer (1903-1964) was one of Pautrier's students, who had been in charge of the Laboratoire d'Histopathologie Cutanee in Strasbourg from 1930 until his death. He achieved a brilliant career in the field of dermatopathology and was very active during the great period between World War I and II. His name is linked to a rare disease, pagetoid reticulosis, which he actually misunderstood, as he was at first convinced that the characteristic intraepidermal changes were due to Paget cells. In this article, we show for the first time the original correspondence between Kolopp (who sent the case to Strasbourg) and Woringer, including discussion with masters such as Pautrier and Civatte. Civatte suggested that these mysterious intraepidermal cells might have common morphologic features with the recently described Sezary cells. After the first publication of pagetoid reticulosis, it took almost 40 years to establish a clear link with mycosis fungoides. It was Braun Falco who named the disease after Woringer and Kolopp in 1974. Woringer would certainly be surprised to know that his name is still world famous, thanks to a rare disease he was the first to extensively describe in 1939, a few days before the declaration of war. PMID- 16314709 TI - Digital papillary adenoma (ADPA) and aggressive digital papillary adenocarcinoma (AdPACa). PMID- 16314711 TI - Sarcomatoid basal cell carcinoma. PMID- 16314713 TI - Slouching toward the lymphatic lumen. PMID- 16314714 TI - A critical analysis of the eccentric starting point for trochanteric intramedullary femoral nailing. AB - OBJECTIVES: Antegrade femoral intramedullary nailing through a greater trochanteric insertion site has been proposed for the treatment of subtrochanteric fractures. The currently available trochanteric nails have dissimilar characteristics, and the most appropriate insertion site for satisfactory subtrochanteric fracture alignment has not been determined. This study is an analysis of 5 different trochanteric femoral nails and 3 different insertion sites using a cadaveric model of a reverse obliquity subtrochanteric femur fracture to determine the optimal trochanteric entry site. SETTING: OSHA approved cadaveric laboratory with an OEC 9800 (General Electric Company, Fairfield, CT) fluoroscopic C-arm. METHODS: Twenty-one embalmed human cadaveric femurs were stripped of soft tissues. Three different starting points on the anteroposterior radiograph were used: at the tip of the greater trochanter, and 2 to 3 mm medial and lateral to the tip. A reverse obliquity subtrochanteric fracture was created. The Trochanteric Antegrade Nail (TAN), Gamma nail (2nd and 3rd generations), Trochanteric Fixation Nail (TFN), and the Holland Nail were then inserted. The proximal bend and radius of curvature were calculated for each nail. Varus and valgus angulation as well as lateral gapping were measured on radiographs; also calculated were the mean, range, and standard deviation. Statistical analysis was performed on angulation and gapping at the fracture site by using Fisher least significant differences analysis, based on a 2-way ANOVA test. RESULTS: The Holland nail had a proximal bend of 10 degrees and a radius of 300 cm. TAN was 5 degrees and 350 cm, TFN was 6 degrees and 150 cm, Gamma 2 was 4 degrees and 300 cm, and Gamma 3 was 4 degrees and 200 cm. The tip starting point led to the most neutral alignment regardless of nail. The lateral starting point led to varus with all nails. The medial starting point led to valgus of >6 degrees with the Holland and TFN; Gamma and TAN had better alignment with <4 degrees of valgus. Gapping of the lateral cortex was greatest with a lateral starting point. CONCLUSIONS: An analysis of 5 trochanteric intramedullary nails with different proximal bends and 3 different starting points in the greater trochanter showed that the tip of the trochanter is close to the "universal" starting point. In this cadaveric subtrochanteric fracture model, the tip starting point led to the most neutral alignment regardless of nail used. The lateral starting point led to varus and gapping of the lateral cortex with all nails. CLINICAL RELEVANCE: Subtrochanteric fractures treated with a trochanteric antegrade nail should have an acceptable reduction before nail insertion. The tip of the trochanter, or even slightly medial, on anteroposterior fluoroscopy is recommended as the universal starting point for these nails. However, slight deviations from this point and nail geometry can cause fracture site malalignment. A lateral starting point led to varus alignment and should be avoided. PMID- 16314715 TI - Treatment of unstable peritrochanteric femoral fractures using a 95 degrees angled blade plate. AB - OBJECTIVE: To clarify the efficacy of a 95-degree angled blade plate fixation in the treatment of unstable peritrochanteric fractures of the femur. DESIGN: Retrospective, clinical study. SETTING: University hospital. PATIENTS: Thirty nine consecutive patients with peritrochanteric femoral fractures were followed for a minimum of 12 months. There were 29 subtrochanteric fractures and 10 intertrochanteric fractures (reverse obliquity pattern) for which the compression hip screw could not be used because of comminution of the trochanteric area and fracture extension to the lateral cortex. The mean age of the patient population at the time of operation was 54 (range, 17-71) years. INTERVENTION: Open reduction and internal fixation of the pertrochanteric fractures using a 95 degrees angled blade plate. MAIN OUTCOME MEASUREMENTS: Time to fracture union, operation time, and complications. RESULTS: Duration of clinical follow-up averaged 26 (range, 12-36) months. The average time to osseous union for those fractures that healed primarily was 19 (range, 13-28) weeks. Two of 39 fractures united with 10 degrees varus deformity, but no corrective surgery was warranted. Limb length discrepancy more than 1.5 cm did not occur. Implant failure before solid bony union occurred in 1 case with a severely comminuted subtrochanteric fracture. Postoperative infection or osteonecrosis of the femoral head did not occur any time throughout the follow-up period. CONCLUSION: A 95 angled blade plate can be a useful alternative fixation device for the treatment of unstable peritrochanteric femoral fractures. PMID- 16314716 TI - Mechanical study of the safe distance between distal femoral fracture site and distal locking screws in antegrade intramedullary nailing. AB - OBJECTIVE: To determine the safe distance for distal femoral fractures relative to the distal locking screws in antegrade intramedullary femoral nailing using a currently available titanium alloy nail design. DESIGN: Cyclic (fatigue) mechanical testing study. SETTING: Biomechanics laboratory. INTERVENTION: Intramedullary nailing of left synthetic fiberglass composite femora with type 32/33-C fractures at 1, 2, 3, and 4 cm from the more proximal of the distal locking screws. MAIN OUTCOME MEASUREMENT: The number of loading cycles to failure of the nail. RESULTS: A load level of 700 N through the femoral mechanical axis was validated as adequate to cause fatigue failure within 200,000 cycles in slotted stainless- steel nails. In the nonslotted titanium alloy nails, this load level caused failure in only 1 of 3 nails with a fracture at 2 cm from the more proximal of the 2 distal locking screws and in 2 of 3 nails with a fracture at 1 cm from the more proximal of the 2 distal locking screws. All of the other nails did not fail >1 million cycles. CONCLUSIONS: Under laboratory conditions, it is safe to assume that an antegrade titanium alloy nail will survive 1 million compression/bending cycles when the fracture is > or = 3 cm from the more proximal of the 2 distal locking screws. PMID- 16314717 TI - Comparative fixation of tibial plateau fractures using alpha-BSM, a calcium phosphate cement, versus cancellous bone graft. AB - OBJECTIVES: To compare the compressive strength of a bone substitute material (alpha-BSM) to cancellous bone when used to fill a defect void in a cadaver model of a Schatzker II split depression fracture of the lateral tibial plateau. DESIGN: Randomized, paired design. SETTING: Biomedical engineering laboratory. PATIENTS: Twenty-six human tibias were harvested from 13 cadavers. Three pairs of tibia fractured during preparation and were excluded. The remaining 10 matched pairs were randomized to fixation by using the bone substitute material or cancellous bone. INTERVENTION: A split depression fracture of the lateral tibial plateau was created in each tibia by using reproducible methods. This fracture was stabilized with a stainless steel L-plate and screws and either alpha-BSM or cancellous bone to fill the defect void. MAIN OUTCOME MEASUREMENTS: Stiffness of the elevated fragment in compression, total depression of the joint at 1000 N. RESULTS: The alpha-BSM bone substitute displayed significantly greater stiffness than cancellous bone constructs in Schatzker II split depression fractures of the lateral tibial plateau (P < 0.0001). Plateau defects displaced significantly less at 1000 N when using alpha-BSM in comparison to cancellous bone (P < 0.0001). CONCLUSIONS: In this cadaveric study, alpha-BSM is an effective bone substitute compared with cancellous bone graft for stabilizing split depression fractures of the lateral tibial plateau. PMID- 16314718 TI - Biomechanical comparison of intramedullary nail and blade plate fixation for tibiotalocalcaneal arthrodesis. AB - OBJECTIVE: Tibiotalocalcaneal arthrodesis is used to manage severe bone loss, arthritis, and/or instability. The goal is to relieve pain through a stable, well aligned hindfoot and ankle. The purpose of this study was 2-fold: to biomechanically compare 1) initial stability, and 2) the effect of bone density on the stability of intramedullary nail and blade plate fixation in tibiotalocalcaneal arthrodesis. DESIGN: Biomechanical study using anatomic specimens. METHODS: Ankle and subtalar joint capsules were exposed for 7 pairs of fresh-frozen anatomic specimens. One ankle from each pair was instrumented with an interlocked intramedullary nail inserted retrograde across the subtalar and ankle joint while the contralateral hind foot was stabilized with a lateral cannulated blade plate. Specimen stability was tested in plantar/dorsiflexion and inversion/eversion to a maximum bending moment of 12 Nm and in internal/external rotation to a maximum torque of 7 Nm. Physical measurements of bone density were made to determine its effect on stability. MAIN OUTCOME MEASURES: Maximum angular displacement of the constructs in plantarflexion, dorsiflexion, inversion, eversion, internal rotation, and external rotation. RESULTS: No significant differences were observed between the plated and nailed constructs in the 3 loading configurations (Power = 0.77). Only 6 pairs were included in the results because of fixation failures. A small but significant reduction in internal rotation alone of 1.8 degrees was found with the plated compared with the nailed construct (P = 0.045). Reduced stability was associated with lower bone density in torsion and inversion/eversion in the plated constructs (r = 0.67- 0.87) with a similar trend seen in torsion in the nailed constructs (r = 0.5). CONCLUSION: Initial construct stabilities and the effect of reduced bone density were found to be similar between the blade plate and the intramedullary nail in tibiotalocalcaneal arthrodesis, thus implant choice may be based on other clinical factors, such as surgeon preference or soft-tissue status. PMID- 16314719 TI - Spica cast application in the emergency room for select pediatric femur fractures. AB - OBJECTIVES: This study was designed to evaluate the efficacy and safety of immediate spica casting in the emergency room (ER) and evaluate the effect of discharge from the emergency room on short- term complications. DESIGN: Retrospective review of patients treated with immediate spica casting in the ER between June 1, 1993 and July 30, 2001. SETTING: Major, pediatric, orthopaedic trauma and referral center. PATIENTS: A total of 145 pediatric femur fractures in children, younger than age 7 years, treated with immediate spica casting in the ER were reviewed to determine radiographic outcome and short-term complication rates. INTERVENTION: All patients underwent immediate spica cast placement in the ER under conscious sedation. Patients meeting specific criteria were discharged immediately from the ER. MAIN OUTCOME MEASUREMENTS: Radiographic acceptability of alignment at fracture union (angulation, shortening), loss of reduction, number of return visits to the emergency room, and clinical outcome at final follow-up. RESULTS: Average follow-up was 20 +/- 16 weeks (range, 1-9 months). Forty-eight patients (33%) were discharged from the ER. No clinical complications were noted at last follow-up. All children younger than age 2 years, and 86.5% of children ages 2 to 5 years, met acceptable malalignment parameters on final radiographs. There were 16 ER visits (11%) for cast problems. Re-reduction in the operating room was needed in 11 patients (8.9%); 6.9% of patients had a cast problem noted during follow-up visits. Only 9% of patients developed a major complication. Initial shortening was the only independent risk factor found to be associated with loss of reduction. Admission status had no significant effect on the number of ER visits or development of a complication. CONCLUSIONS: If there are no associated factors requiring admission (ie, child abuse or polytrauma), spica casting in the ER for pediatric femur fractures followed by immediate discharge can be safely performed with a low complication rate in children younger than age 6 years, nearly eliminating the need for general anesthesia. PMID- 16314720 TI - A comparison between the effects of acetaminophen and celecoxib on bone fracture healing in rats. AB - OBJECTIVES: This study compared the acute treatment effects of systemic analgesics with (celecoxib) and without anti-inflammatory activity (acetaminophen) on bone fracture healing. STUDY DESIGN: Longitudinal time study of fracture healing in rats. METHODS: Closed, mid diaphyseal femur fractures were produced in female Sprague-Dawley rats. The rats were treated for 10 days after fracture with 60 or 300 mg/kg of acetaminophen, 3 or 6 mg/kg of celecoxib, or vehicle by once-daily oral dosing. Fracture healing was measured after 8 weeks by radiographic examination, mechanical testing, and histology. RESULTS: Radiographic scoring indicated that acute celecoxib treatment significantly impaired fracture healing; acetaminophen treatment had no negative effect. Mechanical testing supported the radiographic observations. No negative effects of celecoxib or acetaminophen treatment on the structural properties (peak torque and torsional rigidity) of the healing femurs were detected. In contrast, celecoxib treatment, but not acetaminophen treatment, significantly reduced the material properties (maximum shear stress and shear modulus) of the healing femurs (P < 0.001). Post-mechanical testing examination of the healing femurs found that 73% of the vehicle-treated or acetaminophen-treated femurs had healed as unions (30/41), 27% failed as incomplete unions (11/41), and none failed as nonunions (0%). In contrast, only 21% of the fractured femurs from the celecoxib treated rats had healed as unions (7/34), 53% failed as incomplete unions (18/34), and 26% failed as nonunions (9/34). The proportion of nonunions among the celecoxib-treated rats was significantly higher compared with the control and acetaminophen-treated rats (P < 0.001). Histologic examination indicated that similar to previous studies, celecoxib treatment, but not acetaminophen treatment, altered normal fracture callus morphology in which cartilage rather than new bone abuts the fracture site. CONCLUSIONS: No negative effect from acute acetaminophen treatment on fracture healing was detected. In contrast, acute treatment with celecoxib, a selective cyclooxygenase-2 inhibitor with anti inflammatory activity, significantly impaired fracture healing. PMID- 16314721 TI - Treatment options in pediatric femoral shaft fractures. AB - Fracture of the femur in a pediatric patient presents special problems, and a variety of treatment options. Child abuse and neglect should be considered and evaluated. Fractures in infants (0-18 months) may be treated successfully in a Pavlik Harness. Spica casting is safe and effective in children up to about 6 years or 100 pounds, although complications can occur and careful attention to technique is important. Surgical treatment is superior in most older or larger children or adolescents, and in cases of multiple trauma, soft tissue injury, obesity or head injury. External fixation is minimally invasive, but carries a risk of malunion and refracture. Rigid antegrade intramedully nailing is possible in adolescents of acceptable size, but has a risk of avascular necrosis. Flexible nailing is minimally invasive and well suited to fractures of the central 2/3 of the diaphysis. In comminuted fractures, it may require supplemental external support. Plate fixation is stable and addresses the entire length of the femur. Soft tissue concerns due to surgical exposure can be minimized by the use of submuscular placement technique. PMID- 16314722 TI - Two-incision technique for treatment of complex proximal humerus fractures. AB - The treatment of complex proximal humerus fractures that involve displacement of the greater tuberosity remains challenging. Good functional outcomes and decreased pain are possible with open reduction and internal fixation. Keys to a successful outcome include preservation of blood supply through decreased soft tissue stripping, restoration of normal anatomic relationships (particularly the greater tuberosity), and stable internal fixation that allows early range of motion. To attain these goals, we advocate fixation with a fixed-angle locking proximal humerus plate using a 2-incision approach. A deltopectoral incision is used to expose the humeral shaft and head while the greater tuberosity is visualized by using a lateral incision. The fracture fragments are reduced and the plate is secured by working through these 2 incisions. PMID- 16314723 TI - Tricortical bone graft for primary reconstruction of comminuted distal humerus fractures. AB - A tricortical bone graft harvested from the ilium was used to reconstruct a severely comminuted open intra-articular distal humerus fracture in an adult patient. The patient demonstrated a satisfactory functional and radiologic outcome despite loss of the lateral trochlear lip. When the lateral portion of the fractured trochlea cannot be repaired, excision of the fragments and insertion of an autogenous corticocancellous bone graft from the iliac crest can restore satisfactory function, even in the setting of an open fracture provided there is limited contamination and tissue devitalization. Radiocapitellar contact may be essential to good elbow function in this situation, because the corticocancellous bone graft does not restore the important lateral lip of the trochlea. PMID- 16314724 TI - Minimally invasive exchange tibial nailing for a broken solid nail: case report and description of a new technique. AB - We present a minimally invasive technique for the removal of a broken solid tibial nail. A special device was invented that minimizes the extraction difficulties. After minimal over-reaming, the device is slid in an antegrade fashion over the nail and locked. Retraction is safe and easy. This article is an illustrative case-presentation describing the device and the surgical technique. PMID- 16314725 TI - Acute compartment syndrome of the thigh in a football athlete: a case report and the role of the vacuum-assisted wound closure dressing. AB - We present a case of compartment syndrome of the thigh due to blunt injury in a Division I American football player managed with fasciotomy and vacuum-assisted wound closure. This case report discusses the vacuum-assisted wound closure dressing as an alternative to more traditional closure techniques such as suture retention devices and split-thickness skin grafting. We feel that any surgeon involved in performing fasciotomies should be familiar with this increasingly used closure device and its potential complications. PMID- 16314727 TI - Intertrochanteric osteotomy using a dynamic hip screw for femoral neck nonunion. (J Orthop Trauma 2005;19:329-333). PMID- 16314728 TI - Strategies to reduce hospitalization in the management of heart failure. AB - Progressive and debilitating heart failure (HF) affects almost 5 million, mostly elderly, individuals in the United States. As the elderly population grows in coming decades, the prevalence of HF is expected to increase substantially. In addition to its human toll, HF yields a substantial economic burden, with direct and indirect cost estimates ranging from $27 to $56 billion annually. It is associated with an unacceptably high rehospitalization rate--50% within 6 months- which not only drives burgeoning costs but also provides a signal that current management approaches to HF are less than optimal. Evidence-based treatment approaches, which include the use of beta-blockers, angiotensin-converting enzyme inhibitors, spironolactone, and nesiritide, may offer opportunities for reducing mortality and rehospitalization rates in HF. Yet, because of inadequate discharge guidance and follow-up, many patients with HF are caught in a "revolving door" process that ultimately culminates in exacerbation and rehospitalization. Hospital-based disease management programs have consistently been shown to optimize care and reduce rehospitalization rates in patients with HF. The Hackensack University Medical Center HF program is discussed as an example of a successful HF program. This program represents a multidisciplinary, multifaceted approach to care that emphasizes case management. The core goal of this program is to provide a continuum of care that extends through hospitalization and into the patients' home environment. PMID- 16314732 TI - Interferon-gamma, but not interferon-alpha, induces SOCS 3 expression in human melanoma cell lines. AB - The signal transducers and transcription activators (STATs) and their endogenous inhibitors of the suppressors of cytokine signalling (SOCS) family are major proteins harmonizing the transmission of external signals from the surface membrane to target genes in the nucleus. To correlate the induction of SOCS 3 by interferons (IFNs) on messenger RNA and protein levels with STAT 1 phosphorylation in human malignant melanoma cell lines, we used a unique collection of 18 established malignant melanoma cell lines and six human non malignant normal cells (two melanocytes, two skin keratinocytes and two fibroblasts). IFN-gamma induced SOCS 3 in 83% of melanoma cell lines, whereas IFN alpha stimulated SOCS 3 expression in only 11% of cases. Similarly, melanocytes showed strong induction of SOCS 3 by IFN-gamma and, to a lesser extent, by IFN alpha. In most cases, SOCS 3 expression was paralleled by STAT 1 phosphorylation at tyrosine residues (Y701). In several lines, however, SOCS 3 was not induced despite STAT 1 phosphorylation and, in a few lines, SOCS 3 induction occurred without detectable STAT 1 phosphorylation, indicating that STAT 1 might not be an exclusive inducer of SOCS 3. Similarly, non-malignant cells displayed STAT 1 activation and high levels of SOCS 3 expression after IFN-gamma (but not IFN alpha) treatment. In conclusion, in contrast to IFN-alpha, IFN-gamma appeared to induce SOCS 3 apparently at the transcription level and exhibited higher cytotoxic effects regardless of the cell origin. PMID- 16314733 TI - Effect of inhibition of aloe-emodin on N-acetyltransferase activity and gene expression in human malignant melanoma cells (A375.S2). AB - Arylamine carcinogens and drugs are N-acetylated by cytosolic N-acetyltransferase (NAT), which uses acetyl-coenzyme A as a cofactor. NAT plays an initial role in the metabolism of these arylamine compounds. 2-Aminofluorene is one of the arylamine carcinogens which have been demonstrated to undergo N-acetylation in laboratory animals and humans. Our previous study showed that human cancer cell lines (colon cancer, colo 205; liver cancer, Hep G2; bladder cancer, T24; leukemia, HL-60; prostate cancer, LNCaP; osteogenic sarcoma, U-2 OS; malignant melanoma, A375.S2) displayed NAT activity, which was affected by aloe-emodin in human leukemia cells. The purpose of this study was to determine whether aloe emodin could affect the enzyme activity and gene expression of NAT at the mRNA and protein levels in malignant human melanoma A375.S2 cells. The results showed that aloe-emodin inhibited NAT1 activity (decreased N-acetylation of 2 aminofluorene) in intact cells in a dose-dependent manner. The effect of aloe emodin on NAT1 at the protein level was determined by Western blotting and the mRNA levels were examined by polymerase chain reaction (PCR) and cDNA microarray. These results clearly indicate that aloe-emodin inhibits the mRNA expression and enzyme activity of NAT1 in A375.S2 cells. PMID- 16314734 TI - Differential expression of angioregulatory matricellular proteins in posterior uveal melanoma. AB - Metastases from uveal melanoma, the most common primary malignant eye tumour in adults, develop solely via their vascular bed due to the absence of intraocular lymphatics. The present study investigated the expression in this tumour of three matricellular proteins--Secreted Protein Acidic and Rich in Cysteine (SPARC), thrombospondin 1 (TSP1) and thrombospondin 2 (TSP2)--with putative contrasting roles in the regulation of angiogenesis. Immunohistochemical analysis of the three proteins was carried out in paraffin-embedded specimens from 27 posterior uveal melanomas and was corroborated with Western blot analysis of fresh-frozen samples from seven of the tumours. SPARC immunoreactivity was detected in all specimens and defined two categories of tumour: SPARC-rich (21 of 27 specimens) and SPARC-patchy (six of 27 specimens) uveal melanomas. SPARC-rich tumours had a significantly higher proportion of specimen area occupied by blood vessels (P=0.04) and showed a positive association with the presence of epithelioid-type tumoral cells (P=0.101). TSP1 was not detected by either of the methods in any of the tumours analysed. Some immunopositivity for TSP2 was detected in tumour cells in approximately 40% of specimens, but was not associated with survival, tumour vascularity or any other histopathological indices of survival. The pattern of expression of these matricellular proteins in uveal melanoma is consistent with a cooperative mechanism for establishing an enhanced environment favourable to angiogenesis. Interventions inducing TSP1 expression and/or inhibiting SPARC expression may be candidates for therapies directed towards the inhibition of angiogenesis in posterior uveal melanoma. PMID- 16314735 TI - Five-year results of prognostic value of tyrosinase in peripheral blood of uveal melanoma patients. AB - Tyrosinase-based reverse transcriptase-polymerase chain reaction (RT-PCR) is a method for the detection of circulating melanoma cells in peripheral blood. To our knowledge, no long-term studies on the prognostic impact of tyrosinase PCR in uveal melanoma have yet been reported. In this prospective, non-randomized, observational cohort study, we included 41 patients with uveal malignant melanoma. RT-PCR for tyrosinase was performed in each patient before and after treatment. A clinical follow-up was performed for each patient for at least 5 years, including chest X-ray, serum liver enzyme determination, ultrasound of the liver and bone scintigraphy. The PCR results, age of the patients, tumour size, tumour location, tumour therapy, internal reflectivity, histology, development of distant metastasis and survival rate during follow-up were analysed. At the time of diagnosis, tyrosinase messenger RNA (mRNA) in peripheral blood, suggesting the presence of circulating melanoma cells, was detected in 16 of the 41 patients. Sixty-nine percent of the PCR samples with a positive result prior to therapy revealed a negative result after therapy. The internal reflectivity of the tumour (P=0.021) and the 5-year survival (P=0.023) showed a statistically significant association with positive PCR. It can be concluded that tyrosinase RT-PCR is a sensitive method for the detection of melanoma cells in peripheral blood. This study indicates that the presence of tumour cells in peripheral blood correlates with 5-year survival. Our results suggest a prognostic value of this method. Nevertheless, prospective analysis of a larger cohort is needed to determine the ultimate value of RT-PCR for tyrosinase in blood testing. PMID- 16314736 TI - Relationship between sentinel lymph node status and regression of primary malignant melanoma. AB - The prognostic significance of spontaneous regression of primary melanoma is a controversial issue. Studies on sentinel lymph node status and circulating tumour cells may represent a step towards a better understanding. The clinical details of 269 melanoma patients who underwent sentinel lymph node biopsy were analysed. Correlation was sought between the parameters of the primary tumour, particularly tumours showing a partial intermediate level of regression, and sentinel lymph node status. The presence of circulating tumour cells was studied by reverse transcription-polymerase chain reaction for tyrosinase messenger RNA preoperatively in 94 patients. Of the examined tumours, 27.8% showed histological features of a partial intermediate level of regression. Regressive tumours were localized predominantly on the trunk (P=0.006), were significantly thinner (P<0.0000) and were less frequently ulcerated (P=0.003) than tumours without regression. Moreover, the majority of regressive melanomas were of the superficial spreading type (P<0.0000) and their sentinel node status was more favourable (P=0.026). We demonstrated the presence of circulating tumour cells in five of 26 (19.2%) regressive and 19 of 68 (29.4%) non-regressive tumours. The difference was not significant (P=0.32). By multivariate analysis, however, the Breslow thickness and ulceration of the primary tumour were predictors of the sentinel lymph node status, in agreement with literature data. A partial intermediate level of regression of the primary tumour did not affect unfavourably the sentinel lymph node status in our study. We failed to demonstrate a significant relationship between the presence of circulating tumour cells and either primary tumour regression or the sentinel lymph node status. PMID- 16314737 TI - Decline in angiogenic factors, such as interleukin-8, indicates response to chemotherapy of metastatic melanoma. AB - Serum concentrations of angiogenic factors have been reported to correlate with tumour burden and prognosis in metastatic melanoma. The present study was performed to assess the value of angiogenic factors in serum in indicating response or failure to chemotherapy and immunochemotherapy in stage IV melanoma. Thirty-five patients suffering from stage IV melanoma according to the American Joint Committee on Cancer (AJCC) criteria were included in this prospective study. Before and following chemotherapy or immunochemotherapy, serum levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF-AB), vascular cell adhesion molecule-1 (VCAM 1) and interleukin-8 (IL-8) were measured. Staging examinations following chemotherapy revealed 15 patients with response to therapy (complete response, partial response, stable disease), 14 patients with progressive disease and six patients with mixed response. Patients who responded to therapy showed a significant decrease in the serum level of IL-8 at the time of staging examinations, whereas patients with progressive disease did not. Following chemotherapy, serum concentrations of PDGF-AB had significantly decreased in both patients with response and patients with progressive disease. Comparing the VEGF and bFGF levels of responders and non-responders after a single administration of cytostatics showed significantly lower concentrations in patients with response to therapy. In all patients, a high intra- and inter-individual variability of serum values was observed during application of therapy. It can be concluded that low IL-8 serum levels after chemotherapy indicate response to chemotherapy in stage IV melanoma patients. The persistence of elevated serum levels of VEGF and bFGF following the initial cytostatic administration may help to identify patients resistant to chemotherapy. The distinct variability of serum levels indicates that processes other than tumour angiogenesis also influence the serum concentration of the examined angiogenic factors. PMID- 16314738 TI - Somatostatin receptor scintigraphy with 111In-DOTA-lanreotide and 111In-DOTA-Tyr3 octreotide in patients with stage IV melanoma: in-vitro and in-vivo results. AB - The overexpression of somatostatin receptors (SST-Rs) on various tumour cells provides the molecular basis for the successful use of radiolabelled SST analogues in clinical oncology. The objective of the study was to evaluate the tumour binding of In-1,4,7,10-tetraazacyclo-dodecane-N,N',N'',N'''-tetraacetic acid-lanreotide (In-DOTA-LAN) and In-DOTA-tyrosine-octreotide (In-DOTA-Tyr-OCT) in patients with stage IV melanoma. In addition, we evaluated the potential antiproliferative effect of SST analogues, together with an assessment of the functionality of SST-Rs, on four melanoma cell lines. Twenty-three patients with advanced metastatic melanoma underwent scintigraphy. Thirty-eight of 61 lesions (62%) were positively imaged with In-DOTA-LAN, whereas 23 (37%) were negative. With In-DOTA-Tyr-OCT, 10 of the 23 documented lesions (43%) were positive and 13 (56%) were negative. In vitro, cell lines showed no growth inhibition in the presence of SST analogues and no influence on cell cycle distribution was found with the addition of SST analogues to cultured cells. In addition, no functional surface SST-Rs could be demonstrated on these cell lines. Taken together, our results demonstrate the visualization of metastatic melanoma in a high percentage of patients, probably due to binding of SST analogues to SST-Rs on tumour vessels or infiltrating immune cells. Judging from our data, however, there is no evidence of functional SST-R expression on melanoma cells. PMID- 16314739 TI - Isolated melanoma in the lung where there is no known primary site: metastatic disease or primary lung tumour? AB - Patients with no history of melanoma occasionally present with apparently metastatic melanoma in the lungs, but have no evidence of a primary melanoma. The aims of this study were to investigate the role of surgical resection in the treatment of such patients, and to examine the evidence for a diagnosis of primary pulmonary melanoma in each case. Patients with an unknown primary melanoma who presented with pulmonary disease and subsequently underwent surgical resection were identified from the Sydney Melanoma Unit database. Fifteen patients fulfilled the study criteria. Multiple lesions were present in four. Eight wedge resections and 10 lobectomies were performed. The patients' median survival was 32 months and the 5-year actuarial survival was 42%. This compares with the overall Sydney Melanoma Unit experience of lung resection for melanoma in 83 patients, where the median survival was 19 months and the 5-year survival was 22%. Resection of pulmonary disease in melanoma patients with an unknown primary lesion can result in long-term survival, and even apparent cure. It is possible that some of the patients in this series had primary melanoma of the lung, but this is impossible to prove. PMID- 16314740 TI - Surgical margins in patients with cutaneous melanoma--assessing the adequacy of excision. AB - Appropriate margins of excision for melanoma have been well defined on the basis of prospective clinical trials. Various factors may result in a discrepancy between the intended clinical margin and the pathologic margin ex vivo, however, making it difficult to determine whether adequate excision margins have been obtained. We reviewed the clinical and pathologic margins in 220 patients evaluated at our institution for localized primary cutaneous melanoma between 1995 and 1997. We calculated the percentage difference between the intended clinical margins (C) and the measured pathologic margin (P). We then divided this number by the intended clinical margin to determine the percentage margin discrepancy. We asked whether the margin discrepancy varied as a function of the Breslow depth or anatomic site of the primary lesion, age or sex of the patient, or surgeon performing the excision. Student's t-test was used to determine whether the differences observed between the groups were significant. Pathologic margins were narrower than clinical margins in 59% (129/220) of specimens, equal in 12% (27/220) and wider in 29% (64/220). The median difference between the two measurements was 10% for the group. There was no significant impact of tumor or patient features on margin discrepancy. Margin discrepancy varied by surgeon, but this was not statistically significant. It can be concluded that, in most cases, the discrepancy between the pathologic and clinical margin of a wide excision specimen approximates to 10%. Larger differences should alert the clinician to the possibility of inadequate excision margins. PMID- 16314741 TI - Temozolomide in combination with cisplatin in patients with metastatic melanoma: a phase II trial. AB - The present study was designed to assess the efficacy and safety of combination therapy with temozolomide plus cisplatin in patients with metastatic melanoma. Thirty patients with metastatic melanoma were enrolled. Treatment consisted of intravenous cisplatin (75 mg/m) on day 1 and oral temozolomide (200 mg/m) on days 1 to 5, every 4 weeks. Nine patients (30.0%) achieved an objective response, including two complete (6.7%) and seven partial (23.3%) responses. The median response duration was 161 days. The median progression-free and overall survival times were 72 and 120 days, respectively. Myelosuppression and emesis were the primary toxicities. In conclusion, temozolomide combined with cisplatin is an active and safe first-line chemotherapy regimen with acceptable and easily manageable toxicities in patients with metastatic melanoma. PMID- 16314742 TI - What features do patients notice that help to distinguish between benign pigmented lesions and melanomas?: the ABCD(E) rule versus the seven-point checklist. AB - The ABCD(E) rule and the seven-point checklist are diagnostic aids that have proven to be useful in the hands of physicians; however, little is known of their value to patients with respect to aiding self-detection. The objective of this study was to investigate features that patients notice when identifying melanomas and to explore how well these features correspond to the ABCD(E) rule and the seven-point checklist. A retrospective, modified, case-control study involving patient interviews was performed. All interviews were conducted through the private consulting rooms of a Melbourne dermatologist (JWK) and a Newcastle plastic surgeon (CH) prior to the result of pathology being known to the patients and the interviewers. Sixty-seven patients with benign pigmented skin lesions and 46 patients with melanomas were included. Using a logistic regression model, the change in size/new lesion and change in colour (major criteria, seven-point checklist) were most useful in differentiating between melanomas and benign pigmented lesions in the hands of patients [odds ratio (OR), 4.74; 95% confidence interval (CI), 1.85-12.19; P=0.001; OR, 4.27; 95% CI, 1.62-11.26; P=0.003, respectively). The ABCD(E) rule failed to discriminate between melanoma and other benign pigmented skin lesions. It can be concluded that, of the patients' observations, changes in size or colour were most important in distinguishing between benign pigmented lesions and melanomas. Such features therefore deserve emphasis in public education campaigns. Medical professionals should also remember to seek a history of change in assessing pigmented skin lesions. PMID- 16314743 TI - Age does not appear to be a major indicator of CDKN2A or CDK4 mutations in melanoma patients in Spain. PMID- 16314744 TI - Hypertrophic osteoarthropathy associated with metastatic melanoma. AB - Hypertrophic osteoarthropathy (HOA) is one of the paraneoplastic syndromes most commonly associated with non-small-cell lung cancer. Although pulmonary metastasis is the second most common initial site of melanoma metastasis, HOA is rarely detected in patients with metastatic melanoma in the lung. We report a case of a 45-year-old woman with advanced melanoma who developed HOA after her disease had progressed through first-line systemic therapy. The patient's diagnosis of HOA was made on the basis of digital clubbing, arthralgia, pain, joint effusion and periosteal bone formation on X-ray with negative rheumatologic laboratory studies. Only six cases of HOA in metastatic melanoma have been reported previously. This diagnosis should be considered with lung metastases and the presentation of polyarthralgia with appropriate laboratory and imaging findings. Interestingly, the patient responded to bisphosphonates and second-line chemotherapy with carboplatin and paclitaxel, which is commonly used for lung cancer, not advanced melanoma. As with many paraneoplastic syndromes, successful treatment of the underlying disease was associated with a rapid resolution of the symptoms. PMID- 16314745 TI - Neurological toxicity during metastatic melanoma treatment with fotemustine. AB - Fotemustine is a cytotoxic drug belonging to the group of nitrosourea derivatives, and is used in the treatment of metastatic melanoma, particularly when secondary brain lesions are present. We report the case of a female patient in partial response following 10 courses of fotemustine and featuring a rapidly progressing mental impairment. The clinical and radiological patterns were consistent with a fotemustine-related toxic encephalopathy, as described previously in a recent report. The physician should be aware of this neurological complication of fotemustine, so that it may be recognized early and not attributed erroneously to tumour evolution. PMID- 16314746 TI - The first description of sentinel node biopsy in a patient with amelanotic melanoma of the glans penis. AB - We present the first description of sentinel node biopsy in a patient with amelanotic melanoma of the glans penis. The patient underwent partial amputation of the penis due to tumor of the glans. Pathologic examination of the postoperative specimen revealed the presence of a very rare malignancy- amelanotic melanoma. Sentinel node biopsy, with the use of the combined radiotracer/blue dye technique, was performed. Preoperative lymphoscintigraphy was performed the day before surgery. During surgery, blue dye mapping and intraoperative detection of gamma radiation were used. Two sentinel nodes were identified in the left inguinal region and one sentinel node in the right inguinal region. All sentinel nodes were an intense violet color; in each case, the level of radiation in the sentinel node was almost 20 times higher than the level of radiation in the node bed. Routine hematoxylin and eosin staining and immunohistochemistry (HMB-45) revealed the presence of micrometastasis in one of the sentinel nodes harvested from the left inguinal region. Consequently, left inguinal, iliac and obturatory lymphadenectomies were performed. The final pathologic examination revealed the presence of one metastasis (diameter, 2 mm) in one of the resected non-sentinel nodes. No relapse has been observed during 18 months of follow-up. PMID- 16314749 TI - Lack of association of the thyroid transcription factor 1 gene with mental retardation in the iodine-deficient areas of China. PMID- 16314750 TI - No association between brain-derived neurotrophic factor gene and obsessive compulsive disorder. PMID- 16314751 TI - Mutation analysis of the agouti related protein promoter region and the melanocortin-3 receptor in anorexia nervosa patients. PMID- 16314752 TI - No genetic association between PCSK9 polymorphisms and Alzheimer's disease and plasma cholesterol level in Japanese patients. PMID- 16314754 TI - Autism spectrum disorders in Prader-Willi and Angelman syndromes: a systematic review. AB - Autism spectrum disorders (ASDs) have been linked with maternally derived duplications/triplications of chromosome 15q11-13 and therefore might occur more frequently in people with Prader-Willi syndrome (PWS) when due to uniparental disomy (UPD), than in other forms of chromosomal abnormality involving this region [i.e. deletion (DEL) forms of PWS and DEL+UPD forms of Angelman's syndrome -(AS)]. Twelve studies regarding ASD in PWS and AS were reviewed. It was noteworthy that among the genetically confirmed UPD and DEL cases of PWS and AS, the rate of ASD was 25.3% (38/150; range 0-36.5%) in PWS and 1.9% in AS (2/104; range 0-100%) (Fisher's exact P<0.0001). Among the subset of cases with confirmed UPD or DEL, the rate of ASD in the UPD cases of PWS was significantly higher (20/53) than in the remaining combined samples (i.e. DEL PWS+UPD AS+DEL AS cases; 20/201) (Fisher's exact P<0.0001). ASD in UPD PWS cases (20/53) compared with DEL PWS cases (18/97) was also statistically significant (Fisher's exact P=0.0176). Thus, the limited available evidence supported the prediction that overexpression of maternally imprinted genes in 15q11-13 confers a risk for ASD. Further research will be required to confirm these findings. PMID- 16314755 TI - Lack of association between variations in the melanocortin 5 receptor gene and bipolar disorder. AB - OBJECTIVE: The melanocortin 5 receptor gene maps to the bipolar susceptibility locus on chromosome 18p11.2. Given the biological role of melanocortins and their influence on the hypothalamic-pituitary-adrenal axis, the melanocortin 5 receptor gene is a plausible candidate gene for bipolar disorder. We tested the hypothesis that the potential functional variation Phe209Leu confers susceptibility to bipolar disorder in a case-control study. METHODS: Genotypes for two variations in the coding region and one variation approximately 7 kb upstream from the coding region were obtained from 345 unrelated bipolar I patients and 275 control samples. Genotypes and allele frequencies were compared between groups using chi(2) contingency analysis. RESULTS: Allele frequencies of the Phe209Leu polymorphism did not differ significantly between bipolar patients and controls (P=0.679). Allele frequencies of the C744T and the intergenic A/G polymorphism did not differ significantly between bipolar patients and controls. All variations were in strong linkage disequilibrium. CONCLUSION: Variations in the melanocortin 5 receptor gene are unlikely to confer susceptibility to bipolar disorder in this sample. Further studies are required to elucidate the susceptibility locus for bipolar disorder on chromosome 18p11. PMID- 16314756 TI - Linkage analysis and molecular haplotyping of the dopamine D4 receptor gene promoter region. AB - OBJECTIVES: Polymorphic regions of the dopamine D4 receptor gene and its promoter region are in the focus of psychogenetic association studies. Besides the accurate phenotype characterization, highly reliable genotyping methods are also of outstanding importance in these works. METHODS: DNA samples of 598 healthy unrelated Caucasian individuals were used to validate the described molecular haplotyping methods and to determine the allele, genotype and haplotype frequencies and the linkage disequilibrium between the polymorphisms of the dopamine D4 receptor promoter region. RESULTS: We described a double genotyping system for the -521CT and -616CG polymorphisms, using a polymerase chain reaction restriction fragment length polymorphism or an allele-specific amplification. Allele and genotype frequencies of the novel -615AG single-nucleotide polymorphism are also determined (-615G=13.21%). For molecular haplotyping of the three single-nucleotide polymorphisms and a 120-bp duplication polymorphism, the allele-specific amplification was combined with restriction digestion. The results of the elaborated haplotyping methods were validated by molecular haplotyping of cloned fragments. CONCLUSIONS: The developed methods have been arranged into an 'economic' protocol that might be extended for higher reliability with a double haplotyping ('full mode'). Despite the close proximity of these sites, only a moderate linkage was found between the -615AG and -616CG (Delta(2)=0.162), between the -616AG and -521CT (Delta(2)=0.0221) and between the -615AG and -521CT single-nucleotide polymorphisms (Delta(2)=0.0346). The 120-bp duplication was shown to be in linkage equilibrium with any of the three single nucleotide polymorphisms. Applications of these results should accelerate psychogenetic association studies of the dopamine D4 receptor gene. PMID- 16314757 TI - Apolipoprotein gene E4 allele promoter polymorphisms as risk factors for Alzheimer's disease. AB - Alzheimer's disease is a complex neurodegenerative disorder, characterized by progressive cognitive decline and distinct neuropathology. The apolipoprotein gene E4 allele (APOE 4) is a major risk factor for the disease. Promoter polymorphisms at -491 and -427 may also contribute to the risk. We examined the two polymorphisms in 178 Alzheimer's patients and 141 controls. The -491AA genotype was overrepresented among the patients (68 versus 54%, P=0.01). However, in patients who were APOE4 carriers, the -491AA genotype more than doubled the risk [odds ratio (OR)=2.5; 95% confidence interval (CI)=1.2-5.4], especially in combination with -427TT [odds ratio (OR)=3.3; 95% confidence interval (CI)=1.5 7.7]. Moreover, the -491A/-427T/APOE4/APOC1A haplotype was threefold higher for patients. These results contribute to the evidence that regulation of APOE4 expression modulates risk for Alzheimer's disease. PMID- 16314758 TI - Association of a polymorphism in the Homer1 gene with cocaine dependence in an African American population. AB - OBJECTIVE: While twin and adoption studies have demonstrated that up to 70% of the risk for becoming addicted to cocaine is due to genetic factors, identifying specific genes involved in the development or progression of cocaine dependence has been difficult. The purpose of this study is to determine whether single nucleotide polymorphisms in the Homer1 and Homer2 genes associate with the cocaine-dependent phenotype in an African American population. METHODS: This study utilized a case-control design in which the genotype and allele frequencies for four single-nucleotide polymorphisms in the Homer1 gene and three single nucleotide polymorphisms in the Homer2 gene were compared between African American individuals with a diagnosis of cocaine dependence (n=170) and African American individuals with no history of substance abuse (n=90). RESULTS: The data indicate that one single-nucleotide polymorphism, rs6871510, located in intron 1 of the Homer1 gene significantly (P=0.029) associates with cocaine dependence at the genotype level, and trends toward a significant association at the allele frequency level (chi=2.62, df=1, P=0.106, OR=1.71). None of the single-nucleotide polymorphisms analyzed in the Homer2 gene associates with cocaine dependence. CONCLUSIONS: The results of this study suggest that a polymorphism in the Homer1 gene, rs6871510, is a potential risk factor for the development of cocaine dependence in an African American population, whereas polymorphisms in the Homer2 gene are not. PMID- 16314759 TI - Clozapine-induced weight gain: a study in monozygotic twins and same-sex sib pairs. AB - To assess the relative contribution of genetic factors in antipsychotic-induced weight gain, we explored the similarity in body mass index (BMI) (kg/m(2)) change under clozapine only (clozapine DeltaBMI) and upon additional inclusion of BMI change under prior antipsychotic medication (total DeltaBMI) of five monozygotic twins in comparison with seven same-sex sibs. Twin and sib pairs were identified by a telephone screening of 786 office-based psychiatrists. Measured data on weight and other clinical variables were obtained cross-sectionally and retrospectively from medical records. We found greater similarity in total DeltaBMI in monozygotic twins (intrapair difference 2.78+/-3.41 kg/m(2)) than in same-sex sibs (5.55+/-4.35 kg/m(2)), resulting in heritability estimates of h(2)=0.8 and A=0.45 (ACE twin model). However, intrapair differences in clozapine DeltaBMI were similar between twins (4.18+/-4.27 kg/m(2)) and sibs (4.68+/-4.88 kg/m(2)). We hypothesize that the weight plateau achieved under clozapine is influenced by genetic factors. The weight gain achieved during pretreatment with other antipsychotics seems to limit clozapine-induced weight gain, thus presumably explaining why heritability/similarity in monozygotic twins in comparison with same-sex sibs is greater for total DeltaBMI than for clozapine DeltaBMI. An important caveat is that, owing to the sample size, the heritability estimates have a large standard error and thus have to be interpreted with caution. PMID- 16314760 TI - McLeod syndrome: life-long neuropsychiatric disorder due to a novel mutation of the XK gene. AB - A 50-year-old man presented with worsening, virtually lifelong, chorea and progressive behavioural disturbance, involving disinhibition and hoarding, over 10 years. Clinical assessment revealed chorea, dysarthria, areflexia, an inappropriately jovial, impulsive manner and neuropsychological evidence of frontosubcortical dysfunction. Investigation results included an elevated creatine kinase, caudate atrophy and hypoperfusion, acanthocytes in the peripheral blood and the McLeod phenotype. DNA studies demonstrated a single-base deletion at position 172 in exon 1 of the XK gene, giving rise to a premature stop codon at position 129 in exon 2. PMID- 16314761 TI - A functional prodynorphin promoter polymorphism and opioid dependence. AB - OBJECTIVES: The prodynorphin gene (PDYN) promoter has a repeat polymorphism that is functionally important in association with substance abuse. We examined this polymorphism for association in our sample of 168 opioid-dependent patients and 122 ethnically and geographically matched controls. METHODS: Patients were selected from university-affiliated residential and non-residential addiction treatment programs in the Philadelphia area. A sample of blood was drawn from consenting individuals and genomic DNA was isolated. Polymerase chain reaction amplification of the PDYN promoter was performed and various genotypes were determined on the basis of differing sizes of the polymerase chain reaction products. The genotype and allele data were analyzed by Fisher's exact test. RESULT: A significant difference in genotype (P<0.0006) and allele (P<10) frequencies was found between the African American and European American populations. We did not detect any significant difference in genotype or allele frequencies between the patients and controls within the European American ethnic group. However, we detected a weak association (P=0.013) when we compared allele frequencies of patients and controls in the African American population. CONCLUSIONS: These data suggest that the PDYN repeat polymorphism should be studied in additional opioid-dependent populations. PMID- 16314762 TI - No association between the tryptophan hydroxylase gene polymorphism and major depressive disorders and antidepressant response in a Korean population. AB - The serotonergic neurotransmitter system has been implicated in major depressive disorder (MDD) and appears to be the target of a variety of antidepressants. Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in serotonin biosynthesis, and selective serotonin re-uptake inhibitors exert their activity enhancing the general serotonergic tone. The goal of this study was to investigate whether the A218C polymorphism of the TPH gene is associated with MDD or antidepressant response. All patients were evaluated at the start and in the eighth week of using the 21-item Hamilton Depression Rating Scale. Genotyping was analyzed with polymerase chain reaction. There were no significant differences in genotypes and allele frequencies between the MDD patients (n = 93) and the control group (n = 127) and in the antidepressant response among TPH gene variants. Results suggest that the A218C polymorphism of the TPH gene does not play a major role in pathogenesis in MDD and does not serve as a modulator of antidepressant activity. PMID- 16314763 TI - Serotonin receptor genes HTR3A and HTR3B are not involved in Gilles de la Tourette syndrome. AB - Serotonin has been implicated in a variety of neuropsychiatric disorders. Serotonergic dysfunction is thought to be involved in the pathophysiology of the Gilles de la Tourette Syndrome (GTS). GTS is characterized by multiple vocal and motoric tics. Among selective, competitive 5-HT(3) receptor antagonists, ondansetron represents a promising drug in GTS treatment. In our study both serotonin receptor subunit genes, HTR3A and HTR3B, were examined for sequence variations in GTS patients. We have analyzed DNA samples from 49 patients by SSCP and dHPLC. In HTR3A, we detected five mutations and in HTR3B the analysis revealed six sequence variants. Statistical analysis rated all variants as probably non-disease-related polymorphisms. Yet a certain effect of the detected variants on the severity of the disease cannot be excluded. PMID- 16314764 TI - Phobic anxiety in late-life in relationship to cognition and 5HTTLPR polymorphism. AB - Anxiety in late-life may functionally impair the older adult. In this study the polymorphic region 5HTTLPR of the SLC6A4 gene was examined in relation to phobic anxiety and cognitive function. Sixty-four community-dwelling older adults were genotyped for the 5HTTLPR polymorphism to examine whether late-life phobias are associated with the short (s) allele and whether cognitive impairment may precipitate phobic behaviors in association with the s allele. Our findings suggested that phobic anxiety symptoms are significantly related to lower cognitive function. However, in this sample we did not detect a significant association between phobic anxiety and the 5HTTLPR genotype. The interaction between age-related changes in cognition and anxiety remain an important area for future studies. PMID- 16314766 TI - Parran Award Lecture: insights into the epidemiology of sexually transmitted diseases from Ro = betacD. PMID- 16314767 TI - Vaginal swabs are the specimens of choice when screening for Chlamydia trachomatis and Neisseria gonorrhoeae: results from a multicenter evaluation of the APTIMA assays for both infections. AB - BACKGROUND: Vaginal swabs were recently U.S. Food and Drug Administration-cleared for detecting Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) using Gen Probe Incorporated's APTIMA COMBO2 Assay (AC2). We assessed the APTIMA CT Assay (ACT) for CT, APTIMA GC Assay (AGC) for GC, and AC2 for both organisms using patient- and clinician-collected vaginal swabs. METHOD: Women attending family planning, obstetrics and gynecology, or sexually transmitted disease (STD) clinics had first-catch urines (FCUs), patient-collected vaginal swabs, clinician collected vaginal swabs, and endocervical swabs tested by ACT, AGC, and AC2. A second endocervical swab and FCU were tested using BD ProbeTec (Becton Dickinson) for CT and GC. We calculated sensitivity and specificity using vaginal swabs to detect CT and GC. RESULTS: Of 1,464 subjects enrolled, 180 had CT and 78 GC. ACT sensitivities and specificities for patient-collected vaginal swabs were 98.3% and 96.5%, respectively; for clinician-collected vaginal swabs, 97.2% and 95.2%, respectively. AGC sensitivities and specificities for patient-collected vaginal swabs were 96.1% and 99.3%, respectively; for clinician-collected vaginal swabs, 96.2% and 99.3%, respectively. AC2 results were similar. If an FCU tested positive for CT or GC, >94% of matching vaginal swabs were positive. Positive endocervical swabs showed slightly less concordance (>90% and >88%, respectively). More infected patients were identified using vaginal swabs than FCUs. With AC2, 171 CT-infected patients were identified using FCUs and 196 using patient-collected vaginal swabs. This difference was more pronounced for CT than for GC. CONCLUSIONS: Vaginal swab specimens allowed sensitive and specific detection of CT and GC in the APTIMA assays. Vaginal swabs identified as many infected patients as endocervical swabs and more than FCUs, and may well be the specimen of choice for screening. PMID- 16314768 TI - Women find it easy and prefer to collect their own vaginal swabs to diagnose Chlamydia trachomatis or Neisseria gonorrhoeae infections. AB - BACKGROUND: Self-collected specimens can be used to screen asymptomatic women for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC). We surveyed women's opinions on ease and preferences as to sampling after collecting their own vaginal swab and urine and a physician collection of vaginal swab and cervical swab. METHODS: In 7 North American cities, a questionnaire was used for women after they participated in a clinical trial of nucleic acid amplification testing of various specimens. A total of 1,090 women consenting to gynecologic sampling for CT and GC (82% of those sampled) volunteered to complete the survey. We analyzed the data for ease of self-collection and preferences for a vaginal swab, urine, or cervical swab. RESULTS: The average age was 26.6 years; 59.6% were black, 25.5% white, 11% Hispanic, 1.9% Asian, and 2% unknown. Thirty-five percent had more than one sex partner in the past 6 months, 84.9% had been previously tested for a sexually transmitted infection (STI), and 49.2% had experienced an STI. A total of 90.4% found it very easy to self-collect a vaginal swab. This was not influenced by age, education, or study site. Seventy-six percent preferred a vaginal swab over a pelvic examination, 60% over a urine collection, and 94% indicated that they would be tested more often if a vaginal swab was available. CONCLUSION: Self-collected vaginal swabs were easy to collect and patients preferred them over urine and cervical swabs. PMID- 16314769 TI - Divergent approaches to partner notification for sexually transmitted infections across the European union. AB - OBJECTIVE: The objective of this study was to characterize partner notification (PN) policy and practice for sexually transmitted infections (STIs) in the European Union (EU) and Norway. GOAL: The goal of this study was to promote European good practice in PN. STUDY: This study consisted of a cross-sectional survey using a structured questionnaire to national STI surveillance leads. RESULTS: Considerable heterogeneity exists in approaches to PN. In 2 countries, PN is compulsory for notifiable STIs; and in 13, it is voluntary. Most countries use patient referral but 5 of 15 also offer provider referral. Activity level varies by country and disease. Etiologic treatment of contacts is most common; 5 usually offer epidemiologic treatment and patient-expedited treatment is unusual. Similarities exist with provision mainly by specialized STI treatment services for bacterial STIs. CONCLUSIONS: This divergence in approach calls for the setting of minimum standards for PN delivery together with mechanisms for establishing and sharing best practice. Further research must identify monitoring systems for inclusion in routine surveillance. PMID- 16314770 TI - Factors associated with self-risk perception for sexually transmitted diseases among adolescents. PMID- 16314771 TI - High prevalence of HIV and sexually transmitted infections among indirect sex workers in Cambodia. AB - OBJECTIVES: The goal of this study was to assess the baseline prevalence of and risk factors for HIV and other sexually transmitted infections (STIs) among beer girls enrolled in a behavioral intervention in Battambang, Cambodia. METHODS: Ninety-two of 114 women participated in baseline interviewing, HIV/STI testing, and STI treatment. Blood specimens were tested for syphilis and HIV infection. Self-administered vaginal swabs were tested for trichomonas, bacterial vaginosis (BV), gonorrhea, and chlamydia infections. RESULTS: HIV prevalence was 26%. STI prevalences were: 14% chlamydia, 12% trichomonas, 3% gonorrhea, and 0% syphilis. The prevalence of BV was 43%. A history of sex work was reported by 82%. Consistent condom use with clients was reported by 39%. Increased number of partners and symptoms of STI were significantly associated with HIV infection. DISCUSSION: These data suggest high sexual risk among beer girls in Cambodia. Targeted and frequent HIV and STI interventions are urgently needed in this population. PMID- 16314772 TI - An intervention to reduce vaginal douching among adolescent and young adult women: a randomized, controlled trial. AB - OBJECTIVE: The objective of this study was to evaluate an intervention to reduce vaginal douching among adolescent and young women who report douching. STUDY: This study consisted of a randomized, controlled trial of 275 primarily black adolescent and young adults aged 14 to 23 years. All women participated in 3 15 minute individualized counseling sessions. The experimental group received interventions based on their stage of readiness for ceasing vaginal douching. The comparison condition emphasized healthy eating and nutrition. The primary outcome measure was douching cessation (i.e., no douching in the preceding 3 months) at the 6-and 12-month assessment; a secondary outcome was progression through the stages of change toward douching cessation. RESULTS: Based on an intention-to treat model, participants assigned to the douching intervention group were significantly more likely to report having stopped douching at 6 months (relative risk [RR], 1.34; 95% confidence interval [CI], 1.03-1.73) and at 12 months (RR, 1.60; 95% CI, 1.28-2.00). At baseline, 89.9% of all women reported no intention to stop douching. Also based on an intention-to-treat model, there were no differences in stage across the 2 groups at 6 months (P = 0.29); however, at 12 months, the difference between the intervention and comparison group was statistically significant (P = 0.008). CONCLUSION: Stage-matched interventions can reduce douching among adolescent and young adult women. PMID- 16314773 TI - Laboratory diagnosis of sexually transmitted infections in estonia in 2001-2002: shortcomings with impact on diagnostic quality and surveillance. AB - OBJECTIVES: The objectives of this study were to comprehensively characterize the range, content, and performance of sexually transmitted infection (STI) testing services in Estonia during the period 2001 to 2002 and to determine if the observed diagnostic laboratory practices and methods adhered to international evidence-based recommendations. STUDY: Survey data, focusing on organization and performance characteristics of STI diagnostics services, were assessed using questionnaires, telephone interviews, and site visits to all responding facilities providing STI diagnostics services in Estonia. Guidelines of international evidence-based recommendations for STI testing were used as references. RESULTS: There were significant shortcomings in STI testing availability and practices. Among all participating laboratories diagnosing STIs, only a minority (n = 16, 28%) offered testing for the full minimum range of relevant STIs in Estonia, i.e., Treponema pallidum, Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis. In addition, because testing methods used were not properly selected, confirmation of several infections in accordance with evidence-based requirements was not possible, which has an impact both on STI diagnostic quality and surveillance. PMID- 16314774 TI - A dose-ranging phase I study of dextrin sulphate, a vaginal microbicide, in HIV negative and HIV-positive female volunteers. AB - DESIGN: This phase I dose-ranging study of 2 weeks of twice-daily dextrin sulphate (DS), a sulphated polymer with in vitro activity against HIV, was designed in 2 parts. Part A was a randomized, placebo-controlled, double-blind, 3 arm trial (DS4%, DS1%, placebo) in HIV-negative women. In part B, HIV-positive women received DS4% and HIV-negative women were randomized to DS4% or no gel. RESULTS: One hundred women were enrolled from 2 sites (London and Antwerp): DS4% (n = 50, 20 of whom were HIV-positive), DS1% (n = 20), placebo (n = 10) and no gel (n = 20). There were no withdrawals related to adverse events and no cases of epithelial disruption. Spotting was reported by 24 women and numbers were significantly higher in all the gel groups (23 of 80), including placebo, compared with no gel (one of 20) (Fisher exact test P = 0.037). There was no evidence of a dose effect. Safety tests showed no evidence of systemic absorption. CONCLUSION: Although there was no clear explanation for the spotting, DS4% was well tolerated. It was decided to proceed with DS4% in an expanded safety study in Africa but to restrict entry to low-risk HIV-negative women, include a no-gel group, and monitor intermenstrual bleeding closely. PMID- 16314775 TI - Herpes simplex virus type 2 (HSV-2) Western blot confirmatory testing among men testing positive for HSV-2 using the focus enzyme-linked immunosorbent assay in a sexually transmitted disease clinic. AB - OBJECTIVE: The objective of this study was to define the positive predictive value (PPV) of the Focus herpes simplex virus type 2 (HSV-2) enzyme-linked immunosorbent assay (ELISA) in a low HSV-2 prevalence population and to develop a new test interpretation algorithm. METHODS: HSV-2 Western blots were performed on sera from male sexually transmitted disease clinic patients testing HSV-2 ELISA positive and used to define a new class of indeterminate HSV-2 ELISA result. HSV 2 Western blots were then prospectively performed on sequential sera with indeterminate HSV-2 ELISAs. RESULTS: Ninety-one (84%) of 108 HSV-2 ELISA-positive sera tested HSV-2 Western blot-positive. Western blot positivity was more common in men without herpes simplex virus type 1 (HSV-1) antibody than in those with HSV-1 antibody (93% vs 76%, P = 0.02) and in men with a history or clinical evidence of genital lesions (88% vs 80%, P = 0.30). Selectively raising the ELISA index value defining HSV-2 positivity from >1.1 to >or=3.0 either among HSV-1 positive men or among those without a history or clinical evidence of genital lesions increased the PPV to >or=93%. Prospective evaluation of an algorithm incorporating HSV-1 serostatus found that 11 of 70 persons with indeterminate HSV 2 ELISAs were Western blot-positive. CONCLUSIONS: Clinicians should consider selectively using a higher index value to define Focus ELISA HSV-2 positivity based on either HSV-1 serostatus or clinical circumstances. PMID- 16314776 TI - Trends in pelvic inflammatory disease hospital discharges and ambulatory visits, United States, 1985-2001. AB - OBJECTIVE: The objective of this study was to describe the estimated trends in incidence of pelvic inflammatory disease (PID) among reproductive-aged women in hospital and ambulatory settings. STUDY: Analyses of PID estimates were performed. Three nationally representative surveys conducted by the National Center for Health Statistics (NCHS): National Hospital Discharge Survey (NHDS), National Hospital Ambulatory Medical Care Survey (NHAMCS), and National Ambulatory Medical Care Survey (NAMCS), were used to obtain the estimates of PID (defined by International Classification of Diseases, 9th Revision codes). National Disease and Therapeutic Index (NDTI) estimates were reviewed for comparison. RESULTS: Rates of hospitalized PID declined 68% overall from 1985 through 2001 (P <0.0001). Ambulatory data support a decrease in PID from 1985 to 2001. From 1995 to 2001, approximately 769,859 cases of acute and unspecified PID were diagnosed annually, 91% in ambulatory settings. CONCLUSIONS: PID has decreased in hospital and ambulatory settings. The expanded national surveys in outpatient and emergency departments provide more complete estimates for PID. Optimal management of PID should target ambulatory settings, where the majority of cases are diagnosed and treated. PMID- 16314777 TI - Immune privilege of the eye and fetus: parallel universes? AB - The eye is an extension of the brain and thus many of its tissues are incapable of regeneration. Ocular inflammation can produce extensive damage to innocent bystander cells leading to blindness. However, the eye possesses multiple strategies to control immune-mediated inflammation-a phenomenon known as immune privilege. The fetus of outbred mammals expresses paternal histocompatibility antigens and represents an allograft. However, the success of placental animals is a testament to the immune privilege of the allogeneic fetus. Extensive evidence suggests that the eye and the fetus employ similar strategies for establishing immune privilege for preserving vision and the unborn respectively. PMID- 16314778 TI - An irradiation-free nonmyeloablative bone marrow transplantation model: importance of the balance between donor T-cell number and the intensity of conditioning. AB - BACKGROUND: Animal allogeneic bone marrow transplantation (BMT) models with nonmyeloablative conditioning regimens have so far required irradiation or antibodies in addition to immunosuppressive drugs for engraftment. Moreover, although it is known that the balance between donor T-cell number and the dose of immunosuppressive drugs would be critical for engraftment, it has not been experimentally clarified in a nonmyeloablative regimen. METHODS: We used C57BL/6 mice as donors and DBA/2 mice as recipients with a nonmyeloablative regimen including fludarabine (Flu) and cyclophosphamide (CPA) without irradiation or antibodies. To determine the adequate doses, we injected recipients with various doses of Flu and CPA, and 2x10 bone marrow cells (BMC) and 5x10 splenocytes (SC). Furthermore, using T-cell-depleted BMC and enriched T cells, we investigated the balance between donor T-cell number and the dose of Flu. RESULTS: Doses of Flu at 150 mg/kg/dayx6 and CPA at 150 mg/kg/dayx2 were most appropriate for engraftment with low mortality. All mice appropriately pretreated and transplanted with both BMC and SC exhibited complete donor chimeras. Donor cell engraftment was not enhanced by any increase of BMC transplanted, and dose escalation of donor T cells but not BMC led to the reduction of Flu dose required for engraftment of donor cells. CONCLUSIONS: We have established a murine nonmyeloablative BMT model in a fully MHC-mismatched combination for donor cell engraftment with complete donor chimerism. Simultaneously, we have quantitatively demonstrated that the balance between donor T-cell number and the dose of immunosuppressive drugs is critical for stable engraftment. PMID- 16314779 TI - Mild hypothermic preservation for transport purposes of the AMC bioartificial liver charged with porcine hepatocytes. AB - BACKGROUND: Preservation conditions play a crucial role during transport of a bioartificial liver (BAL) from the laboratory to the hospital. We assessed the possibility to preserve the AMC-BAL loaded with freshly isolated porcine hepatocytes at mild hypothermic temperatures. METHODS: Two laboratory-scale AMC bioreactors were loaded with 1 billion freshly isolated porcine hepatocytes per experiment (n=6). Bioreactors in the control group were kept for three days at 37 degrees C. Bioreactors in the transport group were kept at 37 degrees C during day 1, at 15 degrees C during day 2, and again at 37 degrees C during day 3. In addition, long-term mild hypothermic preservation periods of 45 and 110 hr at 15 degrees C and 26 degrees C, respectively, were assessed. The effect of mild hypothermic preservation on hepatocytes inside the bioreactors was tested by determination of cell damage parameters, as well as metabolic and hepatocyte specific functions. RESULTS: A 24-hour period of mild hypothermic preservation did not reduce any hepatocyte-specific function. LDH release was significantly higher only at day 2. Albumin production at day 2 and lidocaine elimination at day 3 were significantly higher with glucose consumption and lactate production being significantly lower at both test days. Long-term mild hypothermic preservation had a drastic negative effect on cellular viability and hepatocyte specific function. CONCLUSIONS: Mild hypothermic preservation at temperatures as low as 15 degrees C and for a duration of 24 hr is a feasible method to preserve BAL systems loaded with freshly isolated porcine liver cells and will simplify the logistics of BAL transport from the laboratory to the hospital. PMID- 16314780 TI - Combined lung and liver transplantation: the United States experience. AB - BACKGROUND: Combined transplantation of the lungs and liver is indicated for patients who would not be expected to survive transplantation of either organ alone. No single center has accumulated a significant experience, and as a result the expectations for this operation in the current era are unknown. METHODS: Patients that have undergone combined lung-liver transplantation in the United States were enrolled through the United Network for Organ Sharing Organ Procurement and Transplantation Network database. In addition, the English language literature was searched for additional cases of combined lung-liver transplantation. RESULTS: Eleven patients have undergone combined lung and liver transplantation in the United States at different centers. The 1- and 5-year patient survival rates are of 79% and 63%, respectively, and no patient has required retransplantation. These patient survival rates are equivalent to similar a combined lung-liver case series from the United Kingdom (P=0.37, log rank test) and isolated orthotopic liver transplantation in the United States (P=0.59, log-rank test), and are comparable to patient survival rates following isolated lung transplantation in the United States. CONCLUSIONS: Patient survival of combined lung-liver transplantation is comparable to that of isolated liver and isolated bilateral lung transplantation. This option should be considered for patients with end-stage lung disease and liver disease when transplantation of a single organ transplantation is precluded by severe disease in the other organ system. PMID- 16314781 TI - Dynamics of the circadian blood pressure profiles after renal transplantation. AB - BACKGROUND: Abnormalities of diurnal blood pressure (BP) rhythm ("nondipping") are well-described in dialysis patients, and have prognostic importance. It is controversial whether successful renal transplantation (RTx) improves diurnal BP rhythm. To date, no study has attempted to define and model the evolution of diurnal BP rhythm profiles from dialysis to engraftment, focusing on the immediate (4-6 weeks) and medium-term (>1 year) postengraftment periods. METHODS: To test if kidney transplantation normalizes the BP profile, ambulatory blood pressure monitoring (ABPM) was performed in 20 living related transplants (age, 30.3+/-5.1 years; 11 males, on dialysis for 25.6 months) 1 month preRTx and repeated 1 month and >1 year (ABPM3) after successful RTx. Dipping was defined as a sleep-to-awake ratio>0.92 (for systolic BP) and >0.90 (for diastolic BP). RESULTS: PreRTx only 15% patients were dippers. At 1 month postRTx (creatinine clearance, 65.8 ml/min), all patients were complete nondippers. However, after >1 year postRTx (creatinine clearance, 70.4 ml/min), 40% were now dippers. Most importantly, overall, 30% of the patients improved significantly their circadian rhythm (35.3% of the initial preRTx nondippers). Despite successful renal transplantation, 55% patients maintained unchanged their nondipping profile throughout all three ABPM recordings. The only determinants of "long-term" postRTx circadian rhythm are the contemporary level of the renal function and the baseline, dialysis dipping profile: SBP3 sleep-to-awake ratio is related with serum creatinine3 (r=0.58, P=0.001), creatinine clearance (r=-0.41, P=0.036) and SBP1 sleep-to-awake ratio (r=0.48, P=0.034); similarly DBP3 sleep-to-awake ratio is related with serum creatinine3 (r=0.63, P=0.001), creatinine clearance (r= 0.471, P=0.036) and SBP1 sleep-to-awake ratio (r=0.53, P=0.016). In all, 57% of the variance in dipping status can be attributed and explained by the contribution of renal function and initial circadian variability. CONCLUSIONS: Half of the nondipper dialysis patients maintain a permanently abnormal circadian rhythm, despite successful RTx. In the short term, RTx is associated with a highly abnormal diurnal profile, exclusively related to ciclosporin dose and levels. However, in the longer term, renal transplantation leads to a significant improvement of the circadian blood pressure profile, influenced by the renal function level and by the pretransplantation dipping profile. PMID- 16314782 TI - Posttransplant lymphoproliferative disorder among renal transplant patients in relation to the use of mycophenolate mofetil. AB - BACKGROUND: The introduction of increasingly effective immunosuppressants has raised the question of whether posttransplant lymphoproliferative disorder (PTLD), a complication of immunosuppression, would become more frequent. This study assessed the risk of PTLD in relation to immunosuppression during a period that saw the introduction and eventual market dominance of mycophenolate mofetil (MMF). METHODS: A case-control study was conducted at 23 U.S. transplant centers. All participants received a renal-only transplant on or after July 1, 1995. PTLD cases were reported by centers and confirmed by central review. The United Network for Organ Sharing (UNOS) supplemented case ascertainment and identified controls matched on center, transplant date, and age. Center personnel abstracted risk factor and therapy data for cases and up to four controls per case. Cases and controls were compared, using a matched multivariate analysis, to assess the impact of MMF as one component of triple-therapy adjusted for other drug therapies and known risk factors. RESULTS: Data were collected for 108 PTLD cases and 404 controls. PTLD risk for individuals on triple therapy with MMF was similar to the risk experienced by individuals on triple therapy with no MMF (adjusted odds ratio=1.19; 95% CI 0.55-2.55). There was no dose response relationship between MMF and PTLD risk. CONCLUSIONS: Use of MMF was not associated with an increase in PTLD among patients who received triple immunosuppressive therapy, but an excess in risk as large as 155% or a reduction in risk by as much as 45% cannot be ruled out. PMID- 16314783 TI - Liver graft volume estimation in 100 living donors: measure twice, cut once. AB - Estimation of graft volume (GV) is critical in living donor liver transplantation. This study examines the accuracy of formula-derived GV estimates and compares them to both radiogically-derived estimates and actual measurements. We first compared formula-derived estimates of GV and compared them to actual volumes to provide estimates for both right lobe (RL) and left lateral segment (LLS) GV. We then applied these formulae to a validation cohort. Finally, we evaluated both formula-derived and radiologically-derived estimates by comparing them to actual GV measurements. There is a marginal concordance between formula derived calculation and GV for RL donors, but the error ratio was lower than for radiologic estimates. In contrast, MRI measurements for LLS grafts demonstrated a lower error ratio than formula-derived estimation. Formula-derived estimates of GV should be routinely used in the initial screening of potential living donors as long as their limitations are appreciated. PMID- 16314784 TI - Anti-CD2 monoclonal antibody and tacrolimus in adult liver transplantation. AB - BACKGROUND: Blockade of costimulation and adhesion signaling is an attractive approach to interfere with graft rejection METHODS: Between January 1997 and May 1999, forty adults having benign liver diseases were included in a prospective, randomized study comparing tacrolimus plus low-dose short-term steroids without (n=20, TAC group) or with a 10-day course of antihuman CD2 monoclonal antibody (n=20, BTI group). RESULTS: At day 7, histological rejection expressed by mean Banff scores (2.3+/-1.6 vs. 5.4+/-1.6 in the TAC group; P<0.0001) and incidence of moderate to severe rejection (score>or=6) (0 vs. 10 [50%] in the TAC group; P<0.001) were significantly lower in the BTI group. Rejection was treated in 10% (two patients) of BTI patients during the first 3 months and in 15% during the whole follow-up and in 25% (five patients) of TAC patients (P=NS). None of the BTI-patients presented with an adverse event. Three-month, 1-year, and 5-year actual patient survival rates were 100%, 95%, and 95% in the BTI group and 100%, 100%, and 85% in the TAC group. Graft survival rates were 100%, 90%, and 90% in the BTI group and 95%, 95%, and 80% in the TAC group (P=NS). The mAb had no negative impact on infectious or tumor events. CONCLUSIONS: Antihuman CD2 monoclonal antibody is a safe immunosuppressive drug which has a favorable impact on early immunological follow-up of liver transplanted patients. The antibody had no impact on late patient and graft survival. PMID- 16314785 TI - Predictive value of HLA antibodies and serum creatinine in chronic rejection: results of a 2-year prospective trial. AB - In this large collaborative study, 2,231 transplanted patients with functioning kidneys were tested for HLA antibodies, then examined 2 years later for graft survival. Among 478 patients with antibodies, 15.1% failed in 2 years, compared to 6.8% failure in 1,753 patients without antibodies (P=0.00000002). Cytotoxicity testing correlated better with outcome than flow cytometry or ELISA testing on HLA coated beads, possibly because it detected non-HLA antigens. When the patients were further broken down into those with serum creatinine at the time of testing of 0.5-1.9, 4.4% of antibody patients failed at 2 years, compared to 4.3% of patients without antibodies. This 0.1% difference increased among patients with serum creatinine values of 2.0-2.9 to 17.9%, and among those with 3.0-3.9 to 16.3%. We conclude that HLA antibodies posttransplantation is predictive of subsequent graft failure, and its predictive value can be enhanced among patients with higher serum creatinine values. PMID- 16314786 TI - Proteinuria following a switch from calcineurin inhibitors to sirolimus. AB - BACKGROUND: Sirolimus is an alternative option for kidney transplant patients treated with calcineurin inhibitors (CNI) when renal function is deteriorating. However, the incidence of proteinuria following a switch from CNI to sirolimus has caused concern, and was therefore investigated here. METHODS: In a retrospective study, 68 renal transplant recipients were switched from CNI to sirolimus. Proteinuria was measured using 24-hour urine collection before the switch and collections 3, 6, 12, and 24 months thereafter. In addition, proteinuria was measured in patients who had to be switched back to CNI due to side effects. Survival analyses were performed. RESULTS: Baseline proteinuria was 0.39+/-0.69 g/day in all 68 patients. It increased to a mean 1.44+/-1.90 g/day at 3 months (P<0.001) and remained elevated at 6, 12 and 24 months. When sirolimus was withdrawn after the CNI-sirolimus switch for 19 patients, proteinuria decreased from 1.95+/-2.06 g/day to 0.9+/-1.4 g/day (P<0.05). Proteinuria above 0.3 g/day before the CNI-sirolimus switch correlated significantly with the decrease of renal function thereafter. CONCLUSION: CNI-treated kidney transplant recipients may develop reversible proteinuria when switched to sirolimus. PMID- 16314787 TI - Superior outcomes in renal transplantation after early cyclosporine withdrawal and sirolimus maintenance therapy, regardless of baseline renal function. AB - BACKGROUND: It has become increasingly important to refine therapeutic strategies according to individual patient characteristics. We evaluated the long-term impact of renal function at the time of withdrawing cyclosporine (CsA) in renal allograft recipients receiving sirolimus (SRL), CsA, and steroids (ST). METHODS: At 3 months+/-2 weeks, 430 of 525 patients were eligible to be randomized to remain on triple-therapy (SRL-CsA-ST, n=215) or to have CsA withdrawn (SRL-ST, n=215). Patients were divided into quartiles according to their baseline (last value before randomization) calculated GFR: 45 to 56 ml/min (quartile 2, n=105), >56 to 67 ml/min (quartile 3, n=112), and >67 ml/min (quartile 4, n=107). All data were included (ITT analysis). RESULTS: At 4 years, calculated GFR for SRL-CsA-ST vs. SRL-ST was 22.1 vs. 37.7 ml/min (P=0.017), 38.6 vs. 56.6 ml/min (P<0.001), 50.7 vs. 66.8 ml/min (P=0.006), and 62.7 vs. 71.4 ml/min (P=0.436), for quartiles 1 to 4, respectively. Death censored graft loss ranged from 21.2% vs. 7.7% (SRL-CsA-ST vs. SRL-ST, P=0.092) in quartile 1 to 5.5% vs. 1.9% (P=0.618) in quartile 4. The incidence of death and biopsy-confirmed acute rejection also decreased with increasing baseline GFR, but was not significantly different between treatments. Overall, more patients remained on therapy in the SRL-ST group (46.3% vs. 57.9%, P=0.020). CONCLUSIONS: Early and complete withdrawal of CsA from a combination of SRL, CsA, and steroids was preferable to continuing on this regimen, regardless of baseline renal function. The benefit was most marked in patients with a baseline calculated GFRor=2 g/dl, displayed microcytic aregenerative anemia with low serum iron despite high ferritinemia, consistent with anemia of chronic inflammatory states. Fibrinogen and CRP levels increased in these patients after sirolimus introduction. We subsequently focused our study on eight patients without confounding factors of anemia. Anemia improved in all eight after SRL withdrawal. IL6 and TNFalpha at the nadir of anemia were significantly higher than before SRL introduction and after its withdrawal. Decreases in Hb correlated with increases in proinflammatory cytokine levels in a linear regression model. Unchanged serum IL10 levels measured at the nadir of anemia were discordant with the inflammatory state. CONCLUSIONS: Late introduction of SRL may induce anemia and correlates with biochemical evidence of a chronic inflammatory state possibly due to defective IL10-dependent inflammatory autoregulation. PMID- 16314789 TI - Calcineurin inhibitor withdrawal in stable kidney transplant patients decreases the donor-specific cytotoxic T lymphocyte precursor frequency. AB - BACKGROUND: In a prospective study, calcineurin inhibitors (CNI) were withdrawn in patients two years after kidney transplantation. We questioned whether stopping CNI had an effect on the donor-specific reactivity, as CNI might hinder immune responses leading to graft acceptance. METHODS: We measured the donor specific cytotoxic T lymphocyte (CTL) precursor frequency (CTLpf) in 54 patients before and after withdrawal of CNI. In addition, the T-cell reactivity of PBMC to donor and third-party antigens was tested in MLR, and in IFNgamma-Elispot. Reactivity to tetanus toxoid (TET) was studied as well. RESULTS: Donor-specific CTLpf significantly decreased after CNI withdrawal (P=0.0001). In contrast, no difference was observed in third-party reactive CTLpf, donor and third-party reactive MLR and IFNgamma-Elispot. Proliferative responses and the number of IFNgamma-producing cells to TET also decreased after CNI withdrawal. The decrease in CTLpf correlated with the time between the two blood samples (before and after stopping CNI, P=0.05). This decrease was caused by stopping CNI, because there was no correlation between CTLpf and the duration of the CNI treatment after transplantation. Moreover, the percentage of regulatory T cells in the peripheral blood increased after CNI withdrawal. CONCLUSIONS: We report here that after withdrawal of CNI the donor-specific CTLpf decreases. We hypothesize that CNI suppress regulatory mechanisms that have the potential to down-regulate donor specific CTL responses and reactivity to TET. PMID- 16314790 TI - Abnormal B-cell regulation in highly sensitized patients with sustained serum levels of antibody to HLA class I antigens. AB - BACKGROUND: There are anti-idiotypes in the sera of highly sensitized (HS) patients that stimulate B cells to produce antibody to HLA class I antigens. The purpose of this study is to determine if there is an abnormality in B cell responses to these anti-idiotypes. METHODS: Supernatants from normal and HS B cells exposed to either HLA-like anti-idiotypes or HS sera were tested for IgG and antibody to HLA class I antigens by ELISA and flow beads. RESULTS: When stimulated with HS sera, HS B cells produced antibody to HLA class I antigens (in vitro) (12/12) but normal B cells did not (0/10) (P<0.0001). When HS B cells were stimulated with isolated HLA-like anti-idiotypes, they produced more total IgG in the supernatant (603+/-105 ng/ml vs. 293+/-30 ng/ml; P<0.01) and more IgG1 (67+/ 5.3 ng/ml vs. 32.3+/-5.4 ng/ml; P<0.001) and more IgG3 (33.3+/-9.2 vs. 2.03+/-0.2 ng/ml; P<0.0001) than normal B cells. The proliferative response to HLA-like anti idiotypes was 1285+/-115 cpm from normal B cells and 1020+/-445 from HS B cells (p=NS). CONCLUSIONS: When exposed to HS sera, HS B cells produced antibody to HLA class I antigens and normal B cells did not. When exposed to isolated HLA-like anti-idiotypes, HS B cells produced more total IgG, primarily IgG1 and IgG3 with normal proliferation. This intrinsic abnormality in HS B cells permits antibody to HLA class I antigens to be produced and allows increased amounts of IgG1 and IgG3 to be secreted in the absence of an increase in proliferation. PMID- 16314791 TI - Posttransplant lymphoproliferative disorders after renal transplantation in the United States in era of modern immunosuppression. AB - BACKGROUND: Posttransplant lymphoproliferative disorders (PTLD) still represent a major preoccupation after renal transplantation, even in the most recent years. METHODS: We analyzed the incidence, risk, and prognostic factors of PTLD in a cohort of kidney recipients using the United States Renal Data System. RESULTS: Among 25,127 Medicare patients transplanted between 1996 and 2000, 344 developed a PTLD defined as a non-Hodgkin lymphoma (1.4%). History of pretransplant malignancy (adjusted hazard ratio [AHR]=3.54, CI 2.31-5.43), younger age (AHR=1.91, CI 1.18-3.1), fewer HLA matches (AHR=1.32, CI 1.1-1.59) and treatment by ATG (AHR=1.55, CI 1.2-1.99) and OKT3 (AHR=1.37, CI 1-1.76), especially if given for rejection therapy were associated with an increased risk of PTLD. Mycophenolate and azathioprine were associated with a lower risk of PTLD (AHR=0.6, CI 0.47-0.78 and AHR=0.66, CI 0.46-0.95, respectively). IL2-receptor inhibitors and sirolimus did not modify the risk of PTLD. Patients without induction therapy treated with tacrolimus were at greater risk of lymphoma than those treated with new formulations of cyclosporine and those treated with antimetabolites (mycophenolate and azathioprine) have a lower risk of PTLD than those without. Patients with PTLD had poor survival (64% vs. 80% at 5 years). Older age, pretransplant malignancy and OKT3 were risk factors for death whereas treatment with mycophenolate was associated with a better survival (AHR=0.49, CI=0.28-0.82). CONCLUSIONS: Our study highlights the contribution of patient history and immunosuppression in the risk of PTLD in the era of modern immunosuppression. PMID- 16314792 TI - Intensive care unit extubation does not preclude extrarenal organ recovery from donors after cardiac death. AB - BACKGROUND: We have sought to increase the utilization of both renal and extrarenal organs from donors after cardiac death (DCD), including DCD donors with ICU extubation. METHODS: Extubation occurred in the intensive care unit (ICU; n=15) and operating room (OR; n=5). The charts of donors were reviewed for demographics, cause of death, time of asystole and cold perfusion. Recipient's charts were reviewed for graft function, length of hospitalization, serum creatinine (Cr) at discharge and last follow-up. Peak transaminases, amylase, and lipase for liver and pancreas recipients were also reviewed. Data are presented as means+/-SEM. RESULTS: From December 2002 until December 2004, 20 DCD donors were utilized yielding 34 kidney transplants (33 recipients), five liver (1 liver kidney), and two pancreas (SPK) transplants. Mean follow-up overall is 260 days. ICU extubation occurred in 26/33 (78.8%) kidneys, 3/5(60%) livers and 1/2 (50%) pancreata performed. Time from extubation to asystole was 15.9+/-1.9 min and overall warm ischemia time was 12.5+/-1.0 min. Serum Cr at discharge and at last follow-up for renal grafts are 4.3+/-0.5 and 1.9+/-0.3 mg/dl, respectively. Peak AST and ALT levels after OLTx were 3620+/-951 and 1955+/-266 i.u., respectively. Peak and discharge total bilirubin were 8.1+/-0.9 and 2.5+/-0.5 mg/dl. Length of hospitalization was 9.6+/-1.0 and 15.8+/-2.3 days for kidney and liver recipients, respectively. Both pancreas recipients were insulin free after transplant. CONCLUSIONS: ICU extubation should not eliminate extrarenal organs from consideration and may be preferable to OR extubation by improving family support and eliminating OR staff concerns about their role in end-of-life care. PMID- 16314793 TI - Prevention of apoptosis as a possible mechanism behind improved cryoprotection of hematopoietic cells by catalase and trehalose. AB - BACKGROUND: Our previous in vitro work has shown the usefulness of membrane stabilizers and antioxidants as additives in conventional freezing medium to freeze mouse and human hematopoietic cells. The present work was carried out using murine model to test the in vivo engraftment ability of mouse bone marrow frozen with (test cells) or without (control cells) addition of a combination of trehalose and catalase in the medium containing 10% dimethyl sulfoxide (DMSO). METHODS: Viability, nucleated cell recovery, and progenitor content of revived cells were measured. Freezing efficacy was tested by in vivo assays like colony forming unit-spleen (CFU-S), pre-CFU-S, and short-term engraftment of frozen marrow in irradiated mice. Long-term engraftment ability of frozen marrow was assessed using a Ly5.1-Ly 5.2 chimera model. Levels of apoptosis and reactive oxygen species (ROS) generated in revived cells were estimated. The former by Annnexin V, TUNEL, and DNA laddering and the latter by DCFH-DA probe. RESULTS: Our results show that the combination of catalase and trehalose with 10% DMSO improves freezing efficacy not only in terms of viability, cell recovery, and progenitor content but also by in vivo assays like CFU-S, pre-CFU-S, and short- and long-term engraftment. Both the level of apoptosis and ROS generation were considerably reduced in test set as compared to control set. CONCLUSIONS: We conclude that inclusion of a combination of trehalose and catalase in conventional freezing medium leads to enhanced engraftment potential of cryopreserved mouse bone marrow cells probably by preventing apoptotic cell death. Our observation using animal model may have significant clinical implications. PMID- 16314794 TI - Nutrient-related issues affecting successful experimental orthotopic small bowel transplantation. AB - BACKGROUND: This study tested the effectiveness of a nutrient-rich preservation solution in a small animal model of orthotopic whole small bowel transplantation. METHODS: Lewis rats received syngeneic total orthotopic small bowel graft after cold storage for 6 h. Donor small bowel was flushed vascularly with University of Wisconsin (UW) solution and flushed luminally with UW solution or an amino acid rich (AA) solution as follows: Group 1, no luminal flush; Group 2, UW solution; Group 3, AA solution. Biopsies were taken over 14 days posttransplant; energetics, oxidative stress, neutrophil recruitment and histologic injury were assessed. RESULTS: All animals in Groups 1 and 2 failed to survive 12 h posttransplant due to hemorrhagic shock and fluid loss. In contrast, all animals in Group 3 survived the operation; survival after 14 days was 80% (4/5). In Group 3, full recovery of tissue adenylates (ATP and energy charge) to freshly isolated tissue values occurred within 3 days. Oxidative stress as assessed by malondialdehyde (MDA) levels was low in Group 3 throughout 14 d; Groups 1 and 2 exhibited high oxidative stress over the initial 35 min reperfusion (P<0.05). Neutrophil recruitment (myeloperoxidase activity) was significantly reduced in Group 3 tissues, as was histologic injury (P<0.05 compared to Groups 1 and 2). By day 14, Group 3 exhibited complete mucosal restoration. CONCLUSIONS: The data presented in this communication supports the use of an intraluminal preservation solution that is tailored to the metabolic requirements of the small bowel. PMID- 16314795 TI - Effects of Eurocollins solution as aortic flush for the procurement of human pancreas. AB - BACKGROUND: Belzer solution is considered to be the best preservation media used for pancreas transplantation; however, its high cost accounts for approximately 14.5% of all resources allocated by the Brazilian government toward each pancreatic transplant. The objective of the present study was to test a reduction of Belzer solution during pancreas harvest, thereby lowering procedural cost. METHODS: The patients received pancreas-kidney transplantations during the period from January 2003 to August 2004. Patients were divided into two groups. Patients assigned to Group A (n=30) received only Belzer solution (2 L through the aorta artery), whereas patients in Group B (n=16) were perfused first with 1 L of Eurocollins solution followed by 1 L of Belzer solution. The two groups were assessed for differences in the following clinical parameters: the need for insulin replacement or antifungal and anticytomegalovirus treatment, pancreatitis, acute cellular rejection, graft vascular thrombosis, fistulas, intra-abdominal collection, graft loss, deaths, pancreatic ischemia time, and average hospitalization time. RESULTS: No statistically significant differences were observed in any of the parameters analyzed (P<0.05). The use of Eurocollins solution, followed by Belzer solution during pancreas harvesting, did not result in differences in graft survival or functionality, postsurgical complications, or patient survival and hospitalization time, when compared to the use of Belzer solution alone. CONCLUSIONS: Perfusion with 1 L of Eurocollins solution followed by 1 L of Belzer solution during pancreas harvesting seems to be a simple and efficient alternative for reducing the costs of the harvesting process. PMID- 16314796 TI - Combination treatment with donor-specific transfusions and cyclosporine a induces long-term survival of cardiac allografts in miniature Swine. AB - BACKGROUND: To evaluate whether pretransplant donor-specific transfusions (DST) can induce tolerance to cardiac allografts in large animals, heterotopic cardiac transplants were performed across a class I MHC barrier in inbred miniature swine. METHODS: Experimental animals received two DSTs, each containing 1.4x10 viable peripheral blood mononuclear cells, 14 and 7 days prior to transplantation together with a 12-day course of cyclosporine (CyA) (13 mg/kg IV) starting on postoperative day (POD) 0. RESULTS: Untreated (n=2) and DST-only (n=2) treated control animals rejected between POD 6 and 8. Animals treated with CyA alone (n=3) exhibited graft survival to 53, 52 and 59 days. In contrast, the combination of DST and CyA (n=3) led to stable graft function for >200 days. Long term survivors maintained peripheral CML response against donor antigen. Following DSTs, the donor-specific proliferative response of CD8+ recipient T cells was significantly increased (P=0.011), and a significant number of CD8+ T cells underwent apoptosis (10.1% on POD 0; 5.2% on POD -14; P=0.04). None of the DST-treated animals developed donor-specific antibodies. CONCLUSIONS: These results are the first to demonstrate the ability of DST to induce operational tolerance to cardiac allografts in large animals, and they suggest that peripheral mechanisms of tolerance mediate this effect. PMID- 16314797 TI - Effects of JAK3 inhibition with CP-690,550 on immune cell populations and their functions in nonhuman primate recipients of kidney allografts. AB - BACKGROUND: Janus Kinase (JAK) 3 is a tyrosine kinase essential for proper signal transduction downstream of selected cytokine receptors and for robust T-cell and natural killer cells activation and function. JAK3 inhibition with CP-690,550 prevents acute allograft rejection. To provide further insight into the mechanisms of efficacy, we investigated the immunomodulatory effects of CP 690,550 in vitro and in vivo in nonhuman primates. METHODS: Pharmacodynamic assessments of lymphocyte activation, function, proliferation and phenotype were performed in three settings: in vitro in whole blood isolated from untransplanted cynomolgus monkeys (cynos), in vivo in blood from untransplanted cynos dosed with CP-690,550 for 8 days, and in vivo in blood from transplanted cynos immunosuppressed with CP-690,550. Cell surface activation markers expression, IL 2- enhanced IFN-gamma production, lymphocyte proliferation and immune cell phenotype analyzes were performed with multiparametric flow cytometry. RESULTS: In vitro exposure to CP-690,550 resulted in significant reduction of IL-2 enhanced IFN-gamma production by T-cells (maximum inhibition of 55-63%), T-cell surface expression of CD25 (50% inhibitory concentration (IC50); 0.18 microM) and CD71 (IC50; 1.6 microM), and T-cell proliferative capacities measured by proliferating cell nuclear antigen expression (IC50; 0.87 microM). Similar results were observed in animals dosed with CP-690,550. In addition, transplanted animals displayed significant reduction of NK cell (90% from baseline) and T-cell numbers whereas CD8 effector memory T-cell populations were unaffected. CONCLUSIONS: Potent in vitro and in vivo immunomodulatory effects of the JAK3 inhibitor CP-690,550 likely contribute to its efficacy in the prevention of organ allograft rejection. PMID- 16314798 TI - Regulatory role of host CD8+ T lymphocytes in experimental graft-versus-host disease across a single major histocompatibility complex class II incompatibility. AB - BACKGROUND: CD8+ T cells are known to regulate type 2 helper T cell (Th2) alloreactive immune responses but their mode of activation is unclear. We investigated the role of host CD8+ T cells in experimental Th2-type graft-versus host disease (GVHD) where donor/recipient disparity is restricted to a single major histocompatibility complex (MHC) class II antigen. METHODS: Immunoglobulin (Ig) E serum levels, eosinophilia and lymphoid tissue hyperplasia were compared after injection of bm12 CD4+ T cells in either wild-type or CD8+ T cell-deficient (CD8-/-) C57BL/6 mice. In vitro, we explored effects of the addition of CD8+ T cells from wild-type or IFN-gamma-/- mice in mixed leukocyte cultures prepared with beta2 microglobulin-deficient (beta2m-/-) CD4+ T cells as responders or beta2m dendritic cells as stimulators. RESULTS: HyperIgE resolved after 3 weeks in wild-type hosts whereas it persisted for 6 weeks in CD8-/- hosts. Eosinophil infiltrates in lymph nodes were significantly enhanced in CD8-/- hosts. Increased serum levels of IL-5 and IL-13 in CD8-/- hosts confirmed the enhancement of Th2 type responses in the context of recipient CD8+ T cell deficiency. Hyperplasia of lymph nodes and spleen were similar in both groups, as well as in vivo proliferation of donor CD4+ T cells. In vitro, CD8+ T cell regulation of the alloreactive Th2 response depended on their production of IFN-gamma and did not require expression of beta2m on CD4+ T cells or antigen-presenting cells. CONCLUSIONS: Host CD8+ T cells regulate alloreactive Th2 responses during graft versus-host disease through an IFN-gamma dependent pathway, independently of the recognition of beta2m-associated MHC class I molecules. PMID- 16314799 TI - Long-term acceptance of renal allografts following prenatal inoculation with adult bone marrow. AB - BACKGROUND: The aim was to investigate if intravascular in utero injection of adult bone marrow into swine fetuses could lead to macrochimerism and tolerance to the donor. METHODS: Outbred Yorkshire sows and boars screening negative for MHC allele SLA of MGH miniature swine were bred. A laparotomy was performed on the sows at 50 days gestation to expose the uterus. Bone marrow harvested from SLA miniature swine was T-cell depleted and injected intravascularly into seventeen fetuses. Flow cytometry was performed to detect donor cells (chimerism) in the peripheral blood after birth. Mixed lymphocyte reactions (MLR) and cell mediated lympholysis (CML) assays were used to assess the response to donor MHC. Previously frozen skin grafts from the bone marrow donor were placed on the offspring from the first litter. Donor-matched renal transplant from SLA donors were performed on chimeric swine, with and without a short 12-day course of cyclosporine, and one nonchimeric littermate. RESULTS: Nine inoculated offspring demonstrated donor cell chimerism in the peripheral blood and lymphohematopoietic tissues. All animals with detectable chimerism within the first three weeks were consistently nonreactive to donor MHC in vitro. Animals challenged with donor skin grafts displayed prolonged graft survival without producing antidonor antibodies. All chimeric animals accepted donor-matched kidney allografts, even one without cyclosporine. The kidney in the nonchimeric littermate rejected by day 21. CONCLUSIONS: Transplantation of allogeneic adult bone marrow into immunocompetent fetal recipients resulted in chimerism. In utero inoculation led to operational tolerance to the donor's major histocompatibility antigens and long-term acceptance to organ allografts. PMID- 16314800 TI - CCR5, RANTES and CX3CR1 polymorphisms: possible genetic links with acute heart rejection. AB - BACKGROUND: The inflammation response is modulated by the elaborated chemokine chemokine receptor system, which also plays an important role in the development of acute rejection (AR). In this study, we hypothesized that functional genetic variants of some of these modulatory proteins might influence the outcome of AR. METHODS: In a retrospective analysis of a cohort of heart transplanted patients (n=158), we examined eight polymorphisms in four genes implicated in this inflammatory process: RANTES, CCR5, CCR2 and CX3CR1. On the basis of timing occurrence, AR episodes (grade>or= 3A) were classified in "early" (0-3 months posttransplantation; EAR) or "late" outcomes (4-12 months posttransplantation; LAR). RESULTS: The incidences of EAR and LAR were 57.6% and 41%, respectively. Number of LAR episodes was significantly higher in subjects that have already experienced one or more EAR episodes, as compared to subjects that had no EAR (median [25%-75%]: 4 () vs. 1 [1-2.5] respectively; P<0.0001). Statistical univariate analysis showed that none of the mentioned polymorphisms were correlated with EAR or LAR. However, allele-allele association analysis showed that subjects carrying both the CX3CR1 249I allele and CCR5 No-E haplotypes were significantly at lower risk of experiencing EAR (OR=0.2 [95%-CI=0.1-0.5], P=0.001). In contrast subjects carrying both the CCR5 E haplotype and the RANTES 403A allele were significantly at higher risk to develop LAR (OR=8.1 [95%-CI=2.3 28.7], P=0.002). CONCLUSIONS: This exploratory study in heart transplantation suggests that the outcomes of EAR and LAR episodes may be influenced by genetic variant interactions such as "CX3CR1 249I*CCR5 No-E" and "CCR5 E*RANTES -403A." PMID- 16314801 TI - Recipient-specific tolerance after HLA-mismatched umbilical cord blood stem cell transplantation. AB - BACKGROUND: Lower incidence and severity of acute graft versus host disease (GVHD) has been observed in leukemia patients receiving HLA-mismatched umbilical cord (UCB) transplants. However, despite the increased use of UCB in stem cell transplantation, the mechanisms underlying these favorable outcomes are not well delineated. METHODS: We analyzed antigen specific lymphocyte responses after transplant to determine whether the decreased allogeneic responsiveness of UCB lymphocytes is attributable to pan-unresponsiveness, lymphocyte repressive or recipient-specific tolerance. RESULTS: Circulating lymphocytes collected early (3 months) after UCB transplant demonstrate a less naive phenotype compared with that in the infused graft. Additionally, after transplant, circulating peripheral blood UCB-derived lymphocytes produced normal levels of interferon-gamma and proliferated normally when stimulated with mitogen or third party alloantigen. In contrast, when stimulated with recipient antigen, circulating lymphocytes emerging posttransplant did not proliferate nor produce interferon-gamma. Moreover, analysis of interleukin-4 production revealed a Th2 response to recipient antigens. These data indicate early induction of immune tolerance of naive UCB graft lymphocytes with skewing of transplant recipient-specific immune response towards Th2 cytokine profile. CONCLUSIONS: UCB graft lymphocyte immune naivety and observed early tolerance induction may contribute to the observed favorable GVHD incidence, despite infusion of HLA mismatch grafts in the unrelated allogeneic setting. PMID- 16314802 TI - Effect of FTY720 on chronic cyclosporine nephropathy in rats. AB - BACKGROUND: Long-term treatment with cyclosporine A (CsA) causes tubulointerstitial inflammation and fibrosis in the kidney. To define the role of lymphocytes in this process, the novel lymphocyte-specific inhibitor FTY720 was administered to rats with experimental model of chronic CsA nephropathy. METHODS: Sprague-Dawley rats were treated daily for 4 weeks with CsA (7.5 mg/kg), or both CsA and FTY720 (0.125 mg/kg). The effects of FTY720 on CsA-induced renal injury were evaluated using renal function tests and histopathology, and the expression of mediators of CsA-induced renal injury (osteopontin, transforming growth factor beta1 [TGF-beta1], betaig-h3, and angiotensin II). RESULTS: FTY720 treatment significantly decreased T-lymphocyte accumulation in kidneys compared with CsA treatment alone. FTY720 treatment improved not only CsA-induced renal dysfunction but also renal histopathology, demonstrated by decreased macrophage infiltration and interstitial fibrosis. Increased osteopontin, TGF-beta1, betaig-h3, and angiotensin II expression in CsA-treated rat kidneys were decreased with FTY720 treatment. CONCLUSIONS: FTY720 treatment prevents CsA-induced renal injury. PMID- 16314803 TI - The influence of baseline expression of human decay accelerating factor transgene on graft survival and acute humoral xenograft rejection. AB - BACKGROUND: Transgenic pigs expressing human decay accelerating factor (hDAF) have been widely used as donors in various non-human primate transplant models. Despite the use of similar immunosuppressive protocols, there is marked variation in graft survival among centres. The present study was undertaken to determine whether the level of hDAF expression in the pig kidney correlates with the degree of rejection and duration of graft survival. METHODS: hDAF transgenic pigs were provided from two suppliers: Guelph Imutran Centre (G) and Harlan Sprague Dawley (H). Following a bilateral nephrectomy, a single hDAF pig kidney was implanted in the baboon, which was subsequently treated with conventional immunosuppressive protocols. The pig's contralateral kidney was collected to provide baseline data. The severity of acute humoral xenograft rejection (AHXR) was graded as stage I III. hDAF expression was measured using morphologic analysis comparing the contralateral and grafted kidneys at the endpoint. RESULTS: Baseline hDAF expression in kidneys from pigs provided by supplier G was significantly higher than that from supplier H (P<0.01). Furthermore, the survival of baboons receiving grafts from G pigs was significantly longer than those receiving grafts from H pigs (P<0.05). In addition, reduction of hDAF expression at the endpoint was associated with a higher degree of AHXR. Severe apoptosis or necrosis was found in grafts with AHXR II-III. CONCLUSIONS: Pig kidneys from different suppliers have variable baseline hDAF expression, which may have an influence on graft survival. Reduced expression of hDAF in the terminal graft was associated with the severity of rejection. PMID- 16314804 TI - Heart transplantation in patients with chronic hepatitis B: clinical evolution, molecular analysis, and effect of treatment. AB - We evaluated clinical evolution and hepatitis B virus (HBV) molecular changes in heart recipients with chronic HBV infection before transplantation, and studied the effects of lamivudine treatment in patients who experienced HBV reactivation. Nine patients with chronic HBV infection who underwent heart transplantation were investigated. HBV surface/core-promoter/precore/core regions were sequenced. Prior to transplantation, all nine patients had consistently normal ALT and low HBV-DNA levels. Seven experienced HBV reactivation after transplantation (ALT elevated, HBV-DNA>200.000 cps/ml). Lamivudine treatment was initially effective in all patients; three patients during the second year of treatment developed lamivudine resistance-associated mutations (rt-L180M, rt-M204V) with severe disease reactivation, remitted after switch to adefovir treatment. No other significant HBV mutations were identified in the genomic regions studied. Immune suppression is crucial in the reactivation of previous inactive HBV infection and in the liver disease progression in heart recipients. Preemptive lamivudine treatment could be useful in the early management of these patients. PMID- 16314805 TI - Sirolimus monotherapy: feasible immunosuppression for long-term follow-up of kidney transplantation--a pilot experience. AB - Chronic allograft nephropathy (CAN), cardiovascular mortality, and posttransplant malignancy are complications of conventional immunosuppression after kidney transplantation. The aim was to study feasibility of sirolimus (SRL) maintenance monotherapy in a pilot experience. All patients with SRL monotherapy of at least 6 months follow-up were included. In 19 patients, age 58 (34-74) years, SRL monotherapy was introduced 98.1 (49-193) months after transplantation by withdrawing concomitant immunosuppressants from protocols already including SRL or introducing SRL and withdrawing other immunosuppressants. Follow-up is 20.0 (6 41) months. One patient died from hepatocellular carcinoma, diagnosed before SRL monotherapy, with functioning graft. No rejections occurred. SRL trough concentration was 10.7 (4.6-16.1) microg/L. Creatinine (1.77 [1.0-2.9] mg/dL vs. 1.68 [0.8-3.3] mg/dL after 6 months, 1.97 [0.8-4.6] mg/dL at last follow-up; P=NS). Proteinuria increased tendentially (333 [67-893] vs. 890 [46-4011] mg/day). No significant changes of hemoglobin, triglycerides, or cholesterol occurred. SRL monotherapy late after kidney transplantation is feasible in selected patients. PMID- 16314806 TI - Relationship between plasma phospholipid polyunsaturated fatty acid composition and bone disease in renal transplantation. AB - To investigate the relationship between polyunsaturated fatty acid (PUFA) and bone metabolism in renal transplant patients, plasma phospholipid (PP) PUFA levels, biochemical markers of bone turnover and bone mineral density (BMD) were determined in 22 recipients of a first renal allograft at baseline and after a mean 24.4 month follow-up. A significant increase in PP n-3 PUFA content, in the [n-3 PUFA/ arachidonic acid] ratio and in BMD values was observed, as well as a close correlation between the increase in PP n-3 PUFA content and femoral neck BMD. Multivariate regression analysis showed that BMD improvement was positively related to PP n-3 PUFA variation and baseline PP eicosapentaenoic acid levels, and negatively to PP arachidonic acid modification. Tacrolimus- versus cyclosporine-treated patients demonstrated a significant increase in femoral neck BMD and PP n-3 PUFA content. This is the first longitudinal study showing a link between PP-PUFA composition and bone disease in renal transplantation. PMID- 16314807 TI - Reactivation of herpes simplex virus after liver transplantation. AB - Reactivation of herpes simplex virus (HSV) after liver transplantation occurs but the incidence and clinical significance are unknown. We recruited 12 consecutive male patients listed for liver transplantation and positive for HSV antibody. HSV polymerase chain reaction was performed on mouth and penile swabs before and at days 5 and 10 after transplant. Data were recorded regarding clinical evidence of HSV infection, degree of immune suppression, length of intensive care stay, and use of antiviral agents. Five out of twelve patients (42%) had evidence of oral reactivation after transplant although only one had clinical manifestations. Genital reactivation was not seen. Reactivation did not correlate with immune suppression but duration of intensive care stay was a possible risk factor. Ganciclovir prescribed for cytomegalovirus prophylaxis in one patient was ineffective. Oral reactivation of HSV after liver transplantation is common; although clinical significance appears low in this small series, routine testing could be implemented readily. PMID- 16314808 TI - Delayed-onset neutropenia in a patient receiving rituximab as treatment for refractory kidney transplantation. PMID- 16314809 TI - Use of skin grafting to demonstrate tolerance prior to kidney transplantation without immunosuppression in the recipient of a previous bone marrow transplant: response. PMID- 16314810 TI - Liver transplantation for small solitary cholangiocellular carcinoma in cirrhosis: a medical mistake or a new indication? PMID- 16314811 TI - Diabetic mastopathy in a patient with a functioning pancreas transplant. PMID- 16314812 TI - Posttransplant hepatocellular carcinoma metastasis at a skull trauma site. PMID- 16314813 TI - Use of skin grafting to demonstrate tolerance before kidney transplantation without immunosuppression in the recipient of a previous bone marrow transplant. PMID- 16314814 TI - [HIV infection: now a chronic disease]. PMID- 16314815 TI - [Study of the usefulness of pharmacist consultations for patients on antiretroviral regimens]. AB - OBJECTIVE: Highly active anti-retroviral therapies (HAART) in HIV treatment can result in complex treatment regimens. We surveyed HIV patients followed in the infectious disease department of Saint-Germain-en-Laye Hospital to assess the interest of offering patients a consultation with a pharmacist. METHODS: The 3 part questionnaire enabled us to assess the medical and pharmaceutical information given to the patient, adherence, and risk factors for poor observance. The questionnaire was distributed to all patients, regardless of whether they were receiving treatment. A simple adherence score was computed as well as a score for the risk of poor adherence. RESULTS: Ninety patients returned analyzable questionnaires: 65 (72.2%) thought a pharmaceutical consultation would be useful. They felt it should cover in priority the following subjects: drug interactions (51%), secondary effects (49%), and what to do after forgetting a dose (44%). Treatment was perceived as positive by 82 patients (91%) and tolerated well by 57 (65%). Sixty patients (66.3%) reported that they occasionally forgot a dose, 37 (41.3%) that they regularly did. The results showed good adherence by 61.3% and poor adherence by 38.5%. Risk of non-adherence was significantly associated with three factors: the number of pills to take, the number of daily doses, and the length of the treatment. CONCLUSION: Our survey shows the interest of consultations with pharmacists as a clinical service. By reinforcing the patient's understanding, these can complete and supplement the physician's explanation and instructions on pharmaceutical topics, especially those that could not be addressed during the clinical visit. The main aim of this process is to improve adherence, which is a key element in treatment efficacy. PMID- 16314816 TI - [Adverse effects of antiretroviral treatments]. AB - The efficacy of antiretroviral drugs has improved the prognosis for infection by human immunodeficiency virus (HIV); it is nonetheless accompanied by toxicity specific to these agents. Adverse effects of these treatments are a major cause of poor compliance as well as of the cessation or change of treatment. Knowledge of these effects allows physicians to choose an individualized and effective treatment for each patient, one that is well tolerated and appropriate to the patient's sometimes complex medical history. Although some of these adverse effects can be life-threatening (drug eruptions with nevirapine and abacavir, pancreatitis with didanosine, or lactic acidosis with nucleoside analogs), most are reversible. Lipodystrophies, sometimes combining morphologic and metabolic disorders, are observed in some patients after several years of treatment and appear to increase cardiovascular risk. They are one of the major disadvantages of combined treatments. PMID- 16314817 TI - [Management of cirrhosis complications in HIV patients coinfected with hepatitis B or C virus]. AB - Cirrhosis is a serious complication of viral hepatitis, and its incidence is increasing in HIV patients coinfected with HCV or HBV as they live longer, thanks to effective antiretroviral treatment (Haart). HIV coinfection accelerates the progression of fibrosis in hepatitis. To implement preventive measures, prompt diagnosis of cirrhosis is important, either by liver biopsy or the noninvasive tests for fibrosis now under wide study (FibroTest, FibroScan, etc.). Afterwards, assessment of the severity of cirrhosis and screening for complications are both necessary: testing for liver failure (Child-Pugh and MELD scores), portal hypertension (upper gastrointestinal endoscopy), and hepatocellular carcinoma (ultrasound and alpha fetoprotein assay). Careful consideration of drug prescriptions and possible interactions is essential. Specific treatment for hepatitis B or C virus is possible at this stage of cirrhosis, although more difficult, especially for HCV (results influenced by genotype, additional risk of complications by lactic acidosis or hepatic decompensation). Management of the complications of portal hypertension must be planned, as for those without HIV infection. Treatment of hepatocellular carcinoma is still disappointing, and liver transplantation, although possible in these patients, must be evaluated. PMID- 16314818 TI - Should HIV/HCV coinfected patients with severe hepatitis be treated for hepatitis C. AB - FREQUENT COINFECTION: One third of HIV-infected patients also carry the hepatitis C virus. Liver disease (fibrosis) in hepatitis C progresses faster and is more severe in HIV patients than in non-coinfected patients. HEPATITIS C SHOULD BE TREATED: The prolongation of survival of HIV patients since the introduction of highly active antiretroviral treatments (HAART), the faster progression of HCV related cirrhosis in cases of HIV-HCV coinfection, the increased mortality associated with hepatitis, and the hepatotoxicity of antiretroviral treatments are all arguments in favor of treating hepatitis C in HIV patients. A combination of peginterferon and ribavirin is the treatment of choice for hepatitis C. It has been assessed in patients with HIV-HCV coinfection and showed satisfactory levels of prolonged virologic response. A treatment of 48 weeks is recommended regardless of genotype. Early virologic response is an excellent predictive factor for prolonged response; if no response is observed at 12 weeks, treatment can be stopped. IMPACT OF HAART: Antiretroviral treatment seems to have a positive effect on the course of hepatitis C that exceeds its risk of hepatotoxicity. PMID- 16314819 TI - [Efficacy of peginterferon plus ribavirin for HIV-HCV co-infection. Results of the APRICOT study]. AB - Until the publication in 2004 of the Apricot, ACTG 5071, and ANRS Ribavic trials, data about the treatment of patients coinfected with HIV and HCV were sparse. The international Apricot study included 868 HIV-HCV-coinfected patients naive of all HCV treatments. They were randomized into 3 groups for 48 weeks of hepatitis C treatment by peginterferon alpha-2a + ribavirin, or peginterferon alpha-2a + placebo, or interferon alpha-2a + ribavirin. Six months after treatment stopped, sustained virologic response (the principal efficacy criterion, intention-to treat analysis) was observed in 40% of patients treated by peginterferon + ribavirin (29% for the nonresponsive genotype 1, and 62% for genotypes 2 and 3), 20% of those receiving peginterferon + placebo, and 12% of those in the group of nonpegylated interferon + ribavirin. This study also validated the positive and negative predictive values of viral load analyzed at 12 weeks of treatment; it was best for patients in genotypes 2 and 3. The factors associated with the best virologic response were a non-1 genotype and a low HCV viral load (< 800,000 IU/mL). As pointed out in the guidelines of the last European consensus conference on coinfection, the combination of peginterferon + ribavirin is currently the reference treatment for chronic hepatitis C. It should also be the preferred treatment for patients with HIV-HCV coinfection. PMID- 16314820 TI - [HIV vaccines]. AB - The devastation caused by the HIV pandemic will probably not be quelled until a safe and effective vaccine has been developed that is also affordable and available in the developing countries where the impact of this virus is most severe. Considerable progress has been made over the past several years in the search for a vaccine against HIV: roughly 30 different types are currently being investigated in phase I/II clinical trials. Major scientific challenges nonetheless still remain. An ideal vaccine would elicit both neutralizing antibodies and cytotoxic T lymphocytes (CTL) and, perhaps, mucosal immune response. The precise correlates of immunity have not been defined. Recombinant live vector-based vaccines, lipopeptides, and plasmid DNA have all been shown to induce CTL. The different forms of prime-boost vaccine strategies currently being tested in numerous clinical trials provide reasons for hope. PMID- 16314821 TI - [HIV management in Africa]. AB - Africa is the continent hardest hit by the HIV pandemic with, according to the WHO for 2004, more than 25 million people infected. Although the mortality and socioeconomic consequences of HIV infection have been visible and predictable, only since 2000 has the catastrophic situation in Africa and other developing areas mobilized international political and medical attention, in view of the impressive results from antiretroviral treatments where they are accessible. The intolerable lack of access to medical care and effective treatments has engendered a fusion of energy and will - political and scientific, local and international - to make HIV treatment available in Africa. Access to treatment must be accompanied by creation of national public health programs and establishment of infrastructures and trained teams. Necessary steps including anticipation and prevention of pitfalls in the large-scale use of antiretrovirals, long-term planning, organization of supplies and distribution, ensuring the permanence of financial support, and making appropriate strategic choices. These steps are required to guarantee the future and to prevent an epidemic rebound due to drug-resistant HIV strains. The conditions of international economic and medical aid have been clarified and are now better adapted to real needs. Nonetheless large disparities still exist according to country and region. The initial results of cohorts of treated patients show results similar to those in the industrialized countries and provide encouragement about the future. Results in countries such as Uganda and Senegal show that local political involvement is primordial for long-term success. Access to antiretroviral drugs is an urgent and essential marker of comprehensive management, but the conditions of their use must be taken into account in assessing future projects if we want to change the course of HIV in Africa. PMID- 16314822 TI - Signal transduction in human pancreatic cancer: roles of transforming growth factor beta, somatostatin receptors, and other signal intermediates. AB - Pancreatic cancer is a devastating disease because of the lack of early detection markers and effective treatments. It is the fourth leading cause of cancer related death in western countries, including the United States. The mechanisms of pancreatic cancer progression remain unknown. Transforming growth factor beta (TGF-beta), a multifunctional cytokine, regulates cell growth and differentiation in healthy tissues, yet fails to do so in pancreatic cancer. Alterations of the TGF-beta and TGF-beta receptor/Smad signal transduction pathway have been implicated in pancreatic cancer. Furthermore, both the TGF-beta receptor and Smad proteins interact with a variety of cellular signal pathways, such as the somatostatin receptors (SSTRs), ERK1/2, and Wnt signal transduction cascades. This suggests that pancreatic cancer is a multi-gene-controlled malignancy and that effective treatments for pancreatic cancer should be aimed at multiple targets. In this review, we summarized the major signal intermediates involved in pancreatic cancer signal transduction pathways and specifically discussed how alterations in the regulatory functions of TGF-beta and Smad proteins allow for pancreatic carcinogenesis. PMID- 16314823 TI - Inflammation in periodontal tissues in response to mechanical forces. AB - Orthodontic forces are known to produce mechanical damage and inflammatory reactions in the periodontium and dental pulp, as well as inflammatory mediators, e.g. prostaglandins, interleukin (IL)-1, IL-6, tumor necrosis factor alpha, and receptor activator of nuclear factor kappaB ligand, in the periodontal ligament (PDL) and dental pulp. We have studied the effects of aging on the production of inflammatory mediators in the PDL using in vitro and in vitro methods and found that aging of PDL tissues may be an important factor in the severity of periodontal disease through a higher production of inflammatory mediators in response to mechanical forces. Further, the levels of inflammatory mediators in gingival crevicular fluid, an osmotically mediated inflammatory exudates found in the gingival sulcus, have been shown to be significantly elevated during orthodontic treatment. In order to reduce inflammation, low-level laser therapy has been recently studied in vitro and in vitro by many investigators as a substitute for anti-inflammatory drugs. Clinical and experimental studies have shown that low-level laser irradiation reduces orthodontic post-adjustment inflammation. We believe that orthodontic forces (mechanical forces) may play an important role in periodontal inflammation and that low-level laser therapy may be useful for its inhibition. PMID- 16314824 TI - Surfactant proteins SP-A and SP-D in human health and disease. AB - Surfactant proteins A (SP-A) and D (SP-D) are lung surfactant-associated hydrophilic proteins that have been implicated in surfactant homeostasis and pulmonary innate immunity. They are collagen-containing C-type (calcium dependent) lectins, called collectins, and are structurally similar to mannose binding protein of the lectin pathway of the complement system. Being carbohydrate pattern-recognition molecules, they recognize a broad spectrum of pathogens and allergens via the lectin domain, with subsequent activation of immune cells via the collagen region, thus offering protection against infection and allergenic challenge. SP-A and SP-D have been shown to be involved in viral neutralization, clearance of bacteria, fungi, and apoptotic and necrotic cells, down-regulation of allergic reaction, and resolution of inflammation. Studies on single-nucleotide polymorphism, protein levels in broncho-alveolar lavage, and gene knock-out mice have clearly indicated an association between SP-A and SP-D and a range of pulmonary diseases. In addition, recent studies using murine models of allergy and infection have raised the possibility that the recombinant forms of SP-A and SP-D may have therapeutic potential in controlling pulmonary infection, inflammation, and allergies in humans. PMID- 16314825 TI - Toll-like receptor expression and function in airway epithelial cells. AB - Toll-like receptors (TLRs) belong to a family of transmembrane proteins that can recognize and discriminate a diverse array of microbial antigens. Following their activation by specific ligands, TLRs initiate intracellular signaling cascades that culminate in the activation of transcription factors and ultimately lead to changes in pro-inflammatory gene expression. The TLR family constitutes an important component of the innate immune system and, although most commonly considered to be associated with immune cell responses, TLRs are also known to be functionally expressed on a variety of other cell types. Epithelial cells represent a significant component of the cellular content of the airways. These cells provide both a barrier to infection and an active defense mechanism against invading microbes. The expression and function of TLRs on airway epithelial cells has been an area of increasing interest in the recent past. This review will summarize advances in our understanding of the role of TLRs in airway epithelial cells. PMID- 16314826 TI - Human leukocyte antigens as psoriasis inheritance and susceptibility markers. AB - Psoriasis is a multifactoral and heterogenetically inherited disease. The role of hereditary transmission is supported by familial association, twin studies, and correlation with human leukocyte antigens (HLA). Numerous studies have proved that B13, B17, Cw6, and DR7 antigens are positively associated with psoriasis. Cw6 antigen has been repeatedly indicated to be the most significant marker for the risk prediction of the disease. On the basis of epidemiological studies and HLA analysis, a concept of two distinct disease patterns of psoriasis vulgaris was proposed. In type I psoriasis the disease has an early onset, strong correlation with Cw6, B13, B17, and DR7 antigens, and familiar inheritance. Type II psoriasis has a late onset, weak correlation with HLA antigens, and sporadic familiar occurrence. Both types seem to differ clinically. Moreover, some extended haplotypes were shown to be correlated with the disease, especially with the type I psoriasis. Although a psoriasis susceptibility gene(s) has not been yet identified, a number of candidate genes were studied, with evidence for a major locus located within the major histocompatibility complex (PSORS 1). Cw6 allele is the most extensively investigated candidate gene, but present evidence suggests that it is rather in strong linkage disequilibrium with the PSORS 1 gene than the susceptibility allele itself. This article reviews past and current data on the genetic background of psoriasis with special attention to its correlation with HLA antigens. PMID- 16314827 TI - Mannose-binding lectin enhances the attachment and phagocytosis of mycobacteria in vitro. AB - INTRODUCTION: Phagocytosis is the critical first step in the Mycobacterium (M.) tuberculosis-phagocyte interaction. The process involves microbial ligands and phagocyte surface receptors. It is known that serum mannose-binding lectin (MBL), an innate immune system component, may enhance the uptake of microbes by phagocytic cells and activate the complement system. Since phagocytes are the replicative environment for mycobacteria and, as we described earlier, tuberculosis patients differ from controls in serum MBL level, we asked whether MBL plays a role in promoting M. tuberculosis access to phagocytic cells. MATERIAL/METHODS: To estimate the influence of MBL on the phagocytic process, FITC-labeled Mycobacterium bovis BCG was used as a model bacterium. Neutrophils from healthy individuals were used as phagocytes. Phagocytosis was performed in the presence or absence of recombinant MBL (rMBL; 2 or 20 microg/ml). The activation of complement was determined by dot-blot immune assay with monoclonal antibodies against C5b-C9. RESULTS: We showed that phagocytosis of the bacteria was more intensive in the presence of human rMBL. Both attachment and ingestion of mycobacteria were enhanced when MBL and active complement components (fresh serum) were present in the medium. The dot-blot method showed that the bacteria slightly activated complement by themselves. This effect was enhanced in the phagocyte-bacteria co-cultures containing rMBL. CONCLUSIONS: It is possible that MBL may serve in vivo as one of the factors facilitating the entry of mycobacteria into phagocytes, pathogen spread, and the establishment of infection. PMID- 16314828 TI - Characterization of human hepatocytes isolated from non-transplantable livers. AB - INTRODUCTION: The successful use of hepatocytes depends on a reliable demonstration of the functional and morphological integrity of isolated cells. Herein we investigated whether the isolation and cryopreservation of primary human hepatocytes can compromise cell viability and liver-specific characteristics. MATERIAL/METHODS: Hepatocytes were isolated from encapsulated human liver segments by a modified 2-step perfusion technique. Isolated cells were Percoll-purified, cryopreserved, and stored in liquid nitrogen for 1-12 months. For rapid assessment of fresh and cryopreserve/thawed hepatocyte yield and viability, the cells were stained with trypan blue or labeled with fluorochromes. For immunocytochemical analysis, the cells were labeled with monoclonal antibodies for the presence of the following antigens and chemokines: CD3, CD45Ro, CD45Ra, CD34, CD68, CD90, CD95, CD20, HLA-DR, Ki67, PCNA, Bcl-2, p53, CXCR3, CXCR4, and SDF-1. The cells were tested for several specific functions, such as ureagenesis, energy status, MTT activity, lactate dehydrogenase leakage, and total CYP450 content. RESULTS: Assessment of both freshly isolated (Percoll-purified) and cryopreserved/thawed hepatocytes revealed a low constitutive level of contamination by non-parenchymal cells compared with crude (unpurified) preparations and tissue sections. All viable hepatocytes showed intact morphology and retained CYP450 protein, energy status, and urea synthesis. CONCLUSIONS: Modifications in hepatocyte preparations, such as depletion of dead, damaged, and nonparenchymal cells, improves cell purity, which can be adapted to further evaluation of hepatocyte immunogenicity. These data illustrate the importance and feasibility of human hepatocyte banking. PMID- 16314829 TI - Correlation of osteoprotegerin and sRANKL concentrations in serum and bone marrow of multiple myeloma patients. AB - INTRODUCTION: Recent studies suggest that multiple myeloma (MM) triggers osteoclastogenesis by disrupting the balance between the receptor activator of NF kappaB ligand (RANKL) and osteoprotegerin (OPG), its natural antagonist. MATERIAL/METHODS: Determinations of bone marrow (BM) and serum OPG and sRANKL concentrations were performed in 133 MM patients and 42 healthy subjects by the ELISA method using Osteoprotegerin ELISA and sRANKL ELISA kits. RESULTS: MM patients had elevated serum levels of OPG compared with controls (p<0.0001) and OPG levels were higher in patients with renal failure and patients with hipercalcemia (p<0.001 and p=0.04, respectively). Serum OPG levels correlated with age, serum beta 2-microglobulin, and BM OPG concentrations and did not correlate with the presence of osteolysis or with stage of disease. sRANKL serum levels in MM patients and in controls were not statistically different (p=0.42). In MM patients, serum OPG and sRANKL levels were similar at diagnosis and in the plateau phase of disease. There was a correlation between BM and serum sRANKL concentrations (p<0.001). Median values of the sRANKL/OPG ratio for BM and serum of MM patients were 0.14 and 0.11, respectively. The median value of the sRANKL/OPG ratio for the serum of controls was 0.11. CONCLUSIONS: In 20% of MM patients, serum OPG levels are elevated, and this may be a compensative reaction related to increased bone destruction. There is not statistically significant relationship between sRANKL serum and BM levels and the main clinical and laboratory parameters of the disease. Determination of BM and the serum sRANKL/OPG ratio seems to have no clinical value. PMID- 16314830 TI - Identification of genes differentially expressed in glioblastoma versus pilocytic astrocytoma using Suppression Subtractive Hybridization. AB - Glioblastoma (GBM) is a highly malignant glioma, which has the propensity to infiltrate throughout the brain in contrast to pilocytic astrocytoma (PA) of the posterior fossa, which does not spread and can be cured by surgery. We have used Suppression Subtractive Hybridization to define markers that better delineate the molecular basis of brain invasion and distinguish these tumor groups. We have identified 106 genes expressed in PA versus GBM and 80 genes expressed in GBM versus PA. Subsequent analysis identified a subset of 20 transcripts showing a common differential expression pattern for the two groups. GBM differs from PA by the expression of five genes involved in invasion and angiogenesis: fibronectin, osteopontin, chitinase-3-like-1 (YKL-40), keratoepithelin and fibromodulin. PA differs from GBM by the expression of genes related to metabolism (apolipoprotein D), proteolysis (protease-serine-11), receptor and signal transduction (PLEKHB1 for Pleckstrin-Homology-domain-containing-protein-family-B-member-1), transcription/translation (eukaryotic-translation-elongation-factor-1-alpha1) processes and cell adhesion (SPOCK1 for SPARC/Osteonectin-CWCV-kazal-like-domains proteoglycan). The expression of these genes was confirmed by real-time quantitative RT-PCR and immunohistochemistry. This study highlights the crucial role of brain invasion in GBM and identifies specific molecules involved in this process. In addition, it offers a restricted list of markers that accurately distinguish PA from GBM. PMID- 16314831 TI - Suppression of KCMF1 by constitutive high CD99 expression is involved in the migratory ability of Ewing's sarcoma cells. AB - High CD99 expression levels and rearrangements of the EWS gene with ETS transcription factor genes characterize the Ewing's sarcoma family of tumors (ESFT). CD99 is a cell surface glycoprotein whose engagement has been implicated in cell proliferation as well as upregulation and transport of several transmembrane proteins in hematopoietic cells. In ESFT, antibody ligation of CD99 induces fast homotypic cell aggregation and cell death although its functional role in these processes remains largely unknown. Here, using an RNAi approach, we studied for the first time the consequences of modulated CD99 expression in six different ESFT cell lines, representing the most frequent variant forms of EWS gene rearrangement. CD99 suppression resulted in growth inhibition and reduced migration of ESFT cells. Among genes whose expression changes in response to CD99 modulation, the potassium-channel modulatory factor KCMF1 was consistently upregulated. In a series of 22 primary ESFT, KCMF1 expression levels inversely correlated with CD99 abundancy. Cells forced to express ectopic KCMF1 showed a similar reduction in migratory ability as CD99 silenced ESFT cells. Our results suggest that in ESFT, high CD99 expression levels contribute to the malignant properties of ESFT by promoting growth and migration of tumor cells and identify KCMF1 as a potential metastasis suppressor gene downregulated by high constitutive CD99 expression in ESFT. PMID- 16314832 TI - PPARgamma insufficiency promotes follicular thyroid carcinogenesis via activation of the nuclear factor-kappaB signaling pathway. AB - The molecular genetic events underlying thyroid carcinogenesis are poorly understood. Mice harboring a knock-in dominantly negative mutant thyroid hormone receptor beta (TRbetaPV/PV mouse) spontaneously develop follicular thyroid carcinoma similar to human thyroid cancer. Using this mutant mouse, we tested the hypothesis that the peroxisome proliferator-activated receptor gamma (PPARgamma) could function as a tumor suppressor in thyroid cancer in vivo. Using the offspring from the cross of TRbetaPV/+ and PPARgamma+/- mice, we found that thyroid carcinogenesis progressed significantly faster in TRbetaPV/PV mice with PPARgamma insufficiency from increased cell proliferation and reduced apoptosis. Reduced PPARgamma protein abundance led to the activation of the nuclear factor kappaB signaling pathway, resulting in the activation of cyclin D1 and repression of critical genes involved in apoptosis. Treatment of TRbetaPV/PV mice with a PPARgamma agonist, rosiglitazone, delayed the progression of thyroid carcinogenesis by decreasing cell proliferation and activation of apoptosis. These results suggest that PPARgamma is a critical modifier in thyroid carcinogenesis and could be tested as a therapeutic target in thyroid follicular carcinoma. PMID- 16314833 TI - Frequent epigenetic silencing of the bone morphogenetic protein 2 gene through methylation in gastric carcinomas. AB - Recently, it was reported that exogenous bone morphogenetic protein (BMP)-2 acted as an antiproliferative agent in a variety of cell lines, including normal and cancerous gastric cell lines, indicating that BMP-2 plays an important role during cell growth. However, despite the loss of BMP-2 expression in several cancers, the underlying mechanism remains unknown. Epigenetic silencing through DNA methylation is one of the key steps during carcinogenesis. In this study, we found, through analysis by the methylation-specific polymerase chain reaction technique, CpG island methylation of the BMP-2 promoter region in gastric and colon cancer cell lines. BMP-2 mRNA was found to be activated after 5-aza-2' deoxycytidine treatment of the methylation-positive cells. Moreover, 24 of the 56 (42.9%) gastric cancer tissues exhibited promoter methylation. Immunohistochemical staining revealed that 18 of the 24 (75%) gastric cancer tissues without methylation signals exhibited BMP-2 expression, whereas among 20 cancer tissues with strong methylation signals only four (20%) expressed BMP-2 (P = 0.0003). These findings indicate that BMP-2 methylation is strongly associated with the loss of BMP-2 protein expression in the primary gastric carcinomas. BMP 2 methylation was more often observed in diffuse type (60.7%) than in intestinal type (25%) gastric carcinomas (P = 0.007). Thus, aberrant BMP-2 methylation and the resultant loss of BMP-2 expression may be related to gastric carcinogenesis, particularly in the diffuse type. PMID- 16314834 TI - GRB2-mediated recruitment of GAB2, but not GAB1, to SF-STK supports the expansion of Friend virus-infected erythroid progenitor cells. AB - Friend virus induces the development of erythroleukemia in mice through the interaction of a viral glycoprotein, gp55, with a truncated form of the Stk receptor tyrosine kinase, short form-Stk (Sf-Stk), and the EpoR. We have shown previously that the ability of Sf-Stk to participate in the transformation of Friend virus-infected cells requires the kinase activity and Grb2-binding site of Sf-Stk. Here we show that Grb2 heterozygous mice exhibit decreased susceptibility to Friend erythroleukemia and that expansion of erythroid progenitors in response to infection requires the C-terminal SH3 domain of Grb2. A fusion protein in which the Grb2-binding site in Sf-Stk is replaced by Gab2, supports the growth of progenitors from mice lacking Sf-Stk, whereas a Sf-Stk/Gab1 fusion protein does not. Gab2 is expressed in spleens from Friend virus-infected mice, co immunoprecipitates with Sf-Stk and is tyrosine phosphorylated in the presence of Sf-Stk. Mice with a targeted deletion in Gab2 are less susceptible to Friend erythroleukemia and the expansion of erythroid progenitor cells in response to infection can be rescued by expression of Gab2, but not Gab1. Taken together, these data indicate that a Sf-Stk/Grb2/Gab2 complex mediates the growth of primary erythroid progenitor cells in response to Friend virus. PMID- 16314835 TI - Full-length ADAMTS-1 and the ADAMTS-1 fragments display pro- and antimetastatic activity, respectively. AB - The exact role of a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS-1) and the underlying mechanism of its involvement in tumor metastasis have not been established. We have now demonstrated that overexpression of ADAMTS 1 promotes pulmonary metastasis of TA3 mammary carcinoma and Lewis lung carcinoma cells and that a proteinase-dead mutant of ADAMTS-1 (ADAMTS-1E/Q) inhibits their metastasis, indicating that the prometastatic activity of ADAMTS-1 requires its metalloproteinase activity. Overexpression of ADAMTS-1 in these cells promoted tumor angiogenesis and invasion, shedding of the transmembrane precursors of heparin-binding epidermal growth factor (EGF) and amphiregulin (AR), and activation of the EGF receptor and ErbB-2, while overexpression of ADAMTS-1E/Q inhibited these events. Furthermore, we found that ADAMTS-1 undergoes auto proteolytic cleavage to generate the NH(2)- and COOH-terminal cleavage fragments containing at least one thrombospondin-type-I-like motif and that overexpression of the NH(2)-terminal ADAMTS-1 fragment and the COOH-terminal ADAMTS-1 fragment can inhibit pulmonary tumor metastasis. These fragments also inhibited Erk1/2 kinase activation induced by soluble heparin-binding EGF and AR. Taken together, our results suggest that the proteolytic status of ADAMTS-1 determines its effect on tumor metastasis, and that the ADAMTS-1E/Q and the ADAMTS-1 fragments likely inhibit tumor metastasis by negatively regulating the availability and activity of soluble heparin-binding EGF and AR. PMID- 16314836 TI - Genome-wide DNA copy number alterations in head and neck squamous cell carcinomas with or without oncogene-expressing human papillomavirus. AB - Oncogene-expressing human papillomavirus type 16 (HPV16) is found in a subset of head and neck squamous cell carcinomas (HNSCC). HPV16 drives carcinogenesis by inactivating p53 and pRb with the viral oncoproteins E6 and E7, paralleled by a low level of mutations in TP53 and allelic loss at 3p, 9p, and 17p, genetic changes frequently found in HNSCCs of nonviral etiology. We hypothesize that two pathways to HNSCC exist: one determined by HPV16 and the other by environmental carcinogens. To define the critical genetic events in these two pathways, we now present a detailed genome analysis of HNSCC with and without HPV16 involvement by employing high-resolution microarray comparative genomic hybridization. Four regions showed alterations in HPV-negative tumors that were absent in HPV positive tumors: losses at 3p11.2-26.3, 5q11.2-35.2, and 9p21.1-24, and gains/amplifications at 11q12.1-13.4. Also, HPV16-negative tumors demonstrated loss at 18q12.1-23, in contrast to gain in HPV16-positive tumors. Seven regions were altered at high frequency (>33%) in both groups: gains at 3q22.2-qter, 5p15.2-pter, 8p11.2-qter, 9q22-34.1, and 20p-20q, and losses at 11q14.1-qter and 13q11-33. These data show that HNSCC arising by environmental carcinogens are characterized by genetic alterations that differ from those observed in HPV16 induced HNSCC, and most likely occur early in carcinogenesis. A number of genetic changes are shared in both tumor groups and can be considered crucial in the later stages of HNSCC progression. PMID- 16314837 TI - Gene expression signatures and biomarkers of noninvasive and invasive breast cancer cells: comprehensive profiles by representational difference analysis, microarrays and proteomics. AB - We have characterized comprehensive transcript and proteomic profiles of cell lines corresponding to normal breast (MCF10A), noninvasive breast cancer (MCF7) and invasive breast cancer (MDA-MB-231). The transcript profiles were first analysed by a modified protocol for representational difference analysis (RDA) of cDNAs between MCF7 and MDA-MB-231 cells. The majority of genes identified by RDA showed nearly complete concordance with microarray results, and also led to the identification of some differentially expressed genes such as lysyl oxidase, copper transporter ATP7A, EphB6, RUNX2 and a variant of RUNX2. The altered transcripts identified by microarray analysis were involved in cell-cell or cell matrix interaction, Rho signaling, calcium homeostasis and copper binding/sensitive activities. A set of nine genes that included GPCR11, cadherin 11, annexin A1, vimentin, lactate dehydrogenase B (upregulated in MDA-MB-231) and GREB1, S100A8, amyloid beta precursor protein, claudin 3 and cadherin 1 (downregulated in MDA-MB-231) were sufficient to distinguish MDA-MB-231 from MCF7 cells. The downregulation of a set of transcripts for proteins involved in cell cell interaction indicated these transcripts as potential markers for invasiveness that can be detected by methylation-specific PCR. The proteomic profiles indicated altered abundance of fewer proteins as compared to transcript profiles. Antisense knockdown of selected transcripts led to inhibition of cell proliferation that was accompanied by altered proteomic profiles. The proteomic profiles of antisense transfectants suggest the involvement of peptidyl-prolyl isomerase, Raf kinase inhibitor and 80 kDa protein kinase C substrate in mediating the inhibition of cell proliferation. PMID- 16314838 TI - RhoC GTPase is required for PC-3 prostate cancer cell invasion but not motility. AB - It is projected that in 2005, approximately 220 900 men will be newly diagnosed with carcinoma of the prostate (CaP). Men who are diagnosed with locally advanced or metastatic disease undergo androgen ablation therapy and most will relapse and progress within 18 months. Metastasis to bone is the major clinical concern during CaP progression, as it is associated with intractable pain, bone fracture and paralysis resulting from spinal cord compression. Therefore, an understanding of the key mechanisms involved in CaP cell bone metastasis is vital to development of novel treatments. The Rho GTPases are molecular switches involved in cell survival, motility and invasion. Increased expression of RhoC GTPase is linked to enhanced metastatic potential in multiple cancers; however, the role of RhoC GTPase in CaP metastasis has not been addressed. In the current study, we demonstrate that RhoC GTPase is expressed and active in PC-3 CaP cells. RhoC inhibition, either pharmacologically with C3 exotransferase or molecularly through expression of a dominant-negative RhoC, promotes IGF-I stimulated random motility but decreases in vitro invasion and experimental metastases. Inhibition of RhoC activity results in drastic morphologic changes and alterations in the expression and distribution of focal adhesion-related proteins. These data suggest that RhoC inhibition leads to activation of other GTPases involved in nondirected motility and that expression of active RhoC is required for the invasive phenotype of PC-3 cells. PMID- 16314839 TI - Loss of interleukin-2-dependency in HTLV-I-infected T cells on gene silencing of thioredoxin-binding protein-2. AB - The transition from interleukin-2 (IL-2)-dependent to IL-2-independent growth is considered one of the key steps in the transformation of human T-cell leukemia virus type-I (HTLV-I)-infected T cells. The expression of thioredoxin-binding protein-2 (TBP-2) is lost during the transition of HTLV-I-infected T-cell lines. Here, we analysed the mechanism of loss of TBP-2 expression and the role of TBP-2 in IL-2-dependent growth in the in vitro model to investigate multistep transformation of HTLV-I. CpGs in the TBP-2 gene are methylated in IL-2 independent but not in IL-2-dependent cells. Sequential treatment with 5-aza-2' deoxycytidine and a histone deacetylase inhibitor augmented histone acetylation and TBP-2 expression, suggesting that loss of TBP-2 expression is due to DNA methylation and histone deacetylation. In IL-2-dependent cells, a basal level of TBP-2 expression was maintained by IL-2 associated with cellular growth, whereas TBP-2 expression was upregulated on deprivation of IL-2 associated with growth suppression. Overexpression of TBP-2 in IL-2-independent cells suppressed the growth and partially restored responsiveness to IL-2. Knockdown of TBP-2 caused the IL-2-dependent cells to show partial growth without IL-2. These results suggested that epigenetic silencing of the TBP-2 gene results in a loss of responsiveness to IL-2, contributing to uncontrolled IL-2-independent growth in HTLV-I-infected T-cell lines. PMID- 16314840 TI - CDX2 has tumorigenic potential in the human colon cancer cell lines LOVO and SW48. AB - CDX2 is a Drosophila caudal-related homeobox transcription factor that is important for the establishment and maintenance of intestinal epithelial cells. CDX2 is a marker of colon cancer, with strong staining in up to 90% of colonic adenocarcinomas. CDX2 heterozygous-null mice develop colonic neoplasms, which have suggested that CDX2 is a tumor suppressor. However, CDX2 has not been reported to affect xenograft growth. Furthermore, CDX2 is rarely mutated in colon cancer, which has led to suggestions that it may play only a minor role as a tumor suppressor in colon cancer. To understand the functional contributions of CDX2 to colon cancer, we disrupted CDX2 in LOVO and SW48 human colon cancer cell lines by targeted homologous recombination. Consistent with the literature, disruption of CDX2 enhanced anchorage-dependent cell proliferation. However, homozygous loss of CDX2 led to significant inhibition of anchorage-independent growth in LOVO cells, and cell lethality in SW48 cells. Further analyses revealed that disruption of CDX2 led to anchorage-independent G1 to S growth arrest and anoikis. In vivo xenograft studies confirmed that disruption of CDX2 inhibited LOVO tumor growth. These data demonstrate that CDX2 mediates anchorage independent growth and survival. Thus, CDX2 has tumorigenic potential in the human colon cancer cell lines LOVO and SW48. PMID- 16314841 TI - ASPP1, a common activator of TP53, is inactivated by aberrant methylation of its promoter in acute lymphoblastic leukemia. AB - We have analyzed the regulation and expression of ASPP members, genes implicated in the regulation of the apoptotic function of the TP53 tumor-suppressor gene, in acute lymphoblastic leukemia (ALL). Expression of ASPP1 was significantly reduced in ALL and was dependent on hypermethylation of the ASPP1 gene promoter. Abnormal ASPP1 expression was associated with normal function of the tumor-suppressor gene TP53 in ALL. The analyses of 180 patients with ALL at diagnosis showed that the ASPP1 promoter was hypermethylated in 25% of cases with decreased mRNA expression. Methylation was significantly higher in adult ALL vs childhood ALL (32 vs 17%, P = 0.03) and T-ALL vs B-ALL (50 vs 9%, P = 0.001). Relapse rate (62 vs 44%, P = 0.05) and mortality (59 vs 43%, P = 0.05) were significantly higher in patients with methylated ASPP1. DFS and OS were 32.8 and 33.7% for patients with unmethylated ASPP1 and 6.1 and 9.9% for methylated patients (P < 0.001 y P < 0.02, respectively). On the multivariate analysis, methylation of the ASPP1 gene promoter was an independent poor prognosis factor in ALL patients. Our results demonstrate that decreased expression of ASPP1 in patients with ALL is due to an abnormal methylation of its promoter and is associated with a poor prognosis. PMID- 16314842 TI - EBV EBNA 2 stimulates CDK9-dependent transcription and RNA polymerase II phosphorylation on serine 5. AB - EBNA 2 is one of only five viral genes essential for the infection and immortalization of human B cells by the cancer-associated virus Epstein-Barr virus (EBV). EBNA 2 activates cellular and viral transcription and associates with components of the basal transcription apparatus and a number of coactivators. We provide the first evidence to show that the mechanism of transcriptional activation by EBNA 2 also involves phosphorylation of the C terminal domain (CTD) of RNA polymerase II (pol II). We found that transcriptional activation by EBNA 2 was inhibited by a dominant-negative mutant of the pol II CTD kinase, CDK9, and by low concentrations of the CDK9 inhibitor 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole. Moreover, using chromatin immunoprecipitation assays we demonstrated that EBNA 2 stimulates both pol II recruitment and pol II phosphorylation on serine 5 of the CTD in vivo. These results identify a new step in the transcription cycle that is subject to regulation by a key EBV-encoded transcription factor and highlight CDK9 inhibitors as potential anti-EBV agents. PMID- 16314843 TI - Parallel induction of ATM-dependent pro- and antiapoptotic signals in response to ionizing radiation in murine lymphoid tissue. AB - The ATM protein kinase, functionally missing in patients with the human genetic disorder ataxia-telangiectasia, is a master regulator of the cellular network induced by DNA double-strand breaks. The ATM gene is also frequently mutated in sporadic cancers of lymphoid origin. Here, we applied a functional genomics approach that combined gene expression profiling and computational promoter analysis to obtain global dissection of the transcriptional response to ionizing radiation in murine lymphoid tissue. Cluster analysis revealed a prominent pattern characterizing dozens of genes whose response to irradiation was Atm dependent. Computational analysis identified significant enrichment of the binding site signatures of NF-kappaB and p53 among promoters of these genes, pointing to the major role of these two transcription factors in mediating the Atm-dependent transcriptional response in the irradiated lymphoid tissue. Examination of the response showed that pro- and antiapoptotic signals were simultaneously induced, with the proapoptotic pathway mediated by p53 targets, and the prosurvival pathway by NF-kappaB targets. These findings further elucidate the molecular network induced by IR, point to novel putative NF-kappaB targets, and suggest a mechanistic model for cellular balancing between pro- and antiapoptotic signals induced by IR in lymphoid tissues, which has implications for cancer management. The emerging model suggests that restoring the p53 mediated apoptotic arm while blocking the NF-kappaB-mediated prosurvival arm could effectively increase the radiosensitivity of lymphoid tumors. PMID- 16314844 TI - Tumor suppressor candidate TSSC5 is regulated by UbcH6 and a novel ubiquitin ligase RING105. AB - The region of human chromosome 11p15.5 is linked with Beckwith-Wiedemann syndrome that is associated with susceptibility to Wilms' tumor, rhabdomyosarcoma and hepatoblastoma. TSSC5 (tumor-suppressing subchromosomal transferable fragment cDNA; also known as ORCTL2/IMPT1/BWR1A/SLC22A1L) is located in the region. The expression of TSSC5 and other genes in the region is regulated through paternal imprinting. Mutations and/or reduced expression of TSSC5 have been found in certain tumors. TSSC5 encodes an efflux transporter-like protein with 10 transmembrane domains, whose regulation may affect drug sensitivity, cellular metabolism and growth. Here, we present evidences indicating that RING105, a novel conserved RING-finger protein with a PA (protease-associated) domain and a PEST sequence, is a ubiquitin ligase for TSSC5 that can function in concert with the ubiquitin-conjugating enzyme UbcH6. The polyubiquitin target site on TSSC5 was mapped to a region in the 6th hydrophilic loop. Ectopic expression of RING105 in HeLa cells caused an accumulation of cells during G1 that was not observed with the expression of a form of RING105 in which a residue within the RING finger was mutated to inactivate its ligase activity. UbcH6-RING105 may define a novel ubiquitin-proteasome pathway that targets TSSC5 in mammalian cells. PMID- 16314845 TI - Phosphorylation of neurofibromin by PKC is a possible molecular switch in EGF receptor signaling in neural cells. AB - Children with neurofibromatosis (NF1) typically develop central nervous system (CNS) abnormalities, including aberrant proliferation of astrocytes and formation of benign astrocytomas. The NF1 gene encodes neurofibromin, a Ras-GAP, highly expressed in developing neural cells; the mechanism of regulation of neurofibromin as a Ras-GAP, remains however unknown. We now show that, in response to EGF, neurofibromin is in vivo phosphorylated on serine residues by PKC-alpha, in human, rat, and avian CNS cells and cell lines. EGF-induced PKC phosphorylation was prominent in the cysteine/serine-rich domain (CSRD) of neurofibromin, which lies in the N-terminus and upstream of the Ras-GAP domain (GRD), and this modification significantly increased the association of neurofibromin with actin in co-immunoprecipitations. In addition, we show that Ras activation in response to EGF was significantly lowered when C62B cells overexpressed a construct encoding both CSRD + GRD. Moreover, when PKC-alpha was downregulated, the Ras-GAP activity of CSRD + GRD was significantly diminished, whereas overexpressed GRD alone acted as a weaker GAP and in a PKC-independent manner. Most importantly, functional Ras inhibition and EGF signaling shifts were established at the single cell level in C6-derived cell lines stably overexpressing CSRD + GRD, when transient co-overexpression of Ras and PKC depletion prior to stimulation with EGF-induced mitosis. Taken together, these data provide the first evidence of a functional, allosteric regulation of GRD by CSRD, which requires neurofibromin phosphorylation by PKC and association with the actin cytoskeleton. Our data may suggest a novel mechanism for regulating biological responses to EGF and provide a new aspect for the understanding of the aberrant proliferation seen in the CNS of children with NF1. PMID- 16314846 TI - Transcription factor YY1: structure, function, and therapeutic implications in cancer biology. AB - The ubiquitous transcription factor Yin Yang 1 (YY1) is known to have a fundamental role in normal biologic processes such as embryogenesis, differentiation, replication, and cellular proliferation. YY1 exerts its effects on genes involved in these processes via its ability to initiate, activate, or repress transcription depending upon the context in which it binds. Mechanisms of action include direct activation or repression, indirect activation or repression via cofactor recruitment, or activation or repression by disruption of binding sites or conformational DNA changes. YY1 activity is regulated by transcription factors and cytoplasmic proteins that have been shown to abrogate or completely inhibit YY1-mediated activation or repression; however, these mechanisms have not yet been fully elucidated. Since expression and function of YY1 are known to be intimately associated with progression through phases of the cell cycle, the physiologic significance of YY1 activity has recently been applied to models of tumor biology. The majority of the data are consistent with the hypothesis that YY1 overexpression and/or activation is associated with unchecked cellular proliferation, resistance to apoptotic stimuli, tumorigenesis and metastatic potential. Studies involving hematopoetic tumors, epithelial-based tumors, endocrine organ malignancies, hepatocellular carcinoma, and retinoblastoma support this hypothesis. Molecular mechanisms that have been investigated include YY1-mediated downregulation of p53 activity, interference with poly-ADP-ribose polymerase, alteration in c-myc and nuclear factor-kappa B (NF-kappaB) expression, regulation of death genes and gene products, and differential YY1 binding in the presence of inflammatory mediators. Further, recent findings implicate YY1 in the regulation of tumor cell resistance to chemotherapeutics and immune-mediated apoptotic stimuli. Taken together, these findings provide strong support of the hypothesis that YY1, in addition to its regulatory roles in normal biologic processes, may possess the potential to act as an initiator of tumorigenesis and may thus serve as both a diagnostic and prognostic tumor marker; furthermore, it may provide an effective target for antitumor chemotherapy and/or immunotherapy. PMID- 16314847 TI - Overexpression of regenerating islet-derived 1 alpha and 3 alpha genes in human primary liver tumors with beta-catenin mutations. AB - The Wnt/beta-catenin signaling pathway is activated in many human hepatocellular carcinomas (HCC). We tried to identify the genes involved in carcinogenesis and progression of HCC with beta-catenin mutations. We used PCR-based subtractive hybridization to compare gene expression between malignant and benign components of a human HCC occurring in pre-existing adenoma activated for beta-catenin. Two of the genes identified belong to the Regenerating gene (REG) family. They encode the Regenerating islet-derived 3 alpha (REG3A/HIP/PAP/REG-III) and 1 alpha (REG1A) proteins, both involved in liver and pancreatic regeneration and proliferation. Using siRNA directed against beta-catenin, we demonstrated that REG3A is a target of beta-catenin signaling in Huh7 hepatoma cells. The upregulation of REG3A and REG1A expression is significantly correlated to the beta-catenin status in 42 HCC and 28 hepatoblastomas characterized for their beta catenin status. Thus, we report strong evidence that both genes are downstream targets of the Wnt pathway during liver tumorigenesis. PMID- 16314848 TI - Can intra-osseous transplantation improve results of umbilical cord blood transplantation in adult patients? PMID- 16314849 TI - Pre-emptive granulocyte transfusions enable allogeneic hematopoietic stem cell transplantation in pediatric patients with chronic infections. PMID- 16314850 TI - The use of blood conservation methods in addition to erythropoietin allows myeloablative allogeneic stem cell transplantation without the use of blood products. PMID- 16314851 TI - Reduction of experimental colitis in the rat by inhibitors of glycogen synthase kinase-3beta. AB - The effects of the inhibitors of glycogen synthase kinase-3beta (GSK-3beta), TDZD 8 and SB 415286, which can substantially reduce the systemic inflammation associated with endotoxic shock in vivo, have now been investigated on the acute colitis provoked by trinitrobenzene sulphonic acid (TNBS) in the rat. Administration of the GSK-3beta inhibitor TDZD-8 (0.1, 0.33 or 1.0 mg kg-1, s.c., b.i.d., for 3 days) caused a dose-dependent reduction in the colonic inflammation induced by intracolonic TNBS assessed after 3 days, both as the area of macroscopic involvement and as a score using 0-10 scale. Likewise, following administration of the GSK-3beta inhibitor SB 415286 (0.1, 0.33 or 1.0 mg kg-1, s.c., b.i.d., for 3 days), the extent and degree of the TNBS-provoked colonic inflammation was reduced. Administration of either TDZD-8 or SB 415286 reduced the fall in body weight following challenge with TNBS at each dose level studied. The increase in myeloperoxidase activity, an index of neutrophil infiltration into the TNBS-induced inflamed colon, was significantly inhibited by both TDZD-8 and SB 415286 at each dose level. The increase in the levels of the proinflammatory cytokine, TNF-alpha, in the inflamed colon was also significantly inhibited by either compound at the highest doses evaluated. The elevated levels of the transcription factor NF-kappaB subunit p65, as determined by Western blot in the nuclear extracts from the TNBS-provoked inflamed colonic tissue, were dose dependently reduced by TDZD-8 or SB 415286 treatment. These findings demonstrate that two chemically distinct selective inhibitors of the activity of GSK-3beta reduce the inflammation and tissue injury in a rat model of acute colitis. The mechanisms underlying this anti-inflammatory action may be related to downregulation of NF-kappaB activity, involved in the generation of proinflammatory mediators. PMID- 16314852 TI - Comparison of HERG channel blocking effects of various beta-blockers-- implication for clinical strategy. AB - beta-Blockers are widely used in the treatment of cardiovascular diseases. However, their effects on HERG channels at comparable conditions remain to be defined. We investigated the direct acute effects of beta-blockers on HERG current and the molecular basis of drug binding to HERG channels with mutations of putative common binding site (Y652A and F656C). beta-Blockers were selected based on the receptor subtype. Wild-type, Y652A and F656C mutants of HERG channel were stably expressed in HEK293 cells, and the current was recorded by using whole-cell patch-clamp technique (23 degrees C). Carvedilol (nonselective), propranolol (nonselective) and ICI 118551 (beta(2)-selective) inhibited HERG current in a concentration-dependent manner (IC(50) 0.51, 3.9 and 9.2 microM, respectively). The IC(50) value for carvedilol was a clinically relevant concentration. High metoprolol (beta(1)-selective) concentrations were required for blockade (IC(50) 145 microM), and atenolol (beta(1)-selective) did not inhibit the HERG current. Inhibition of HERG current by carvedilol, propranolol and ICI 118551 was partially but significantly attenuated in Y652A and F656C mutant channels. Affinities of metoprolol to Y652A and F656C mutant channels were not different compared with the wild-type. HERG current block by all beta blockers was not frequency-dependent. Drug affinities to HERG channels were different in beta-blockers. Our results provide additional strategies for clinical usage of beta-blockers. Atenolol and metoprolol may be preferable for patients with type 1 and 2 long QT syndrome. Carvedilol has a class III antiarrhythmic effect, which may provide the rationale for a favourable clinical outcome compared with other beta-blockers as suggested in the recent COMET (Carvedilol Or Metoprolol European Trial) substudy. PMID- 16314853 TI - Regional heterogeneity in the haemodynamic responses to urotensin II infusion in relation to UT receptor localisation. AB - The aim of the study was to measure regional haemodynamic responses to 6 h infusions of human urotensin II (hUII), to identify possible mediators of the effects observed, and to relate the findings to the distribution of urotensin II receptors (UT receptors). Male, Sprague-Dawley rats had pulsed Doppler flow probes and intravascular catheters implanted for measurement of regional haemodynamics in the conscious, freely moving state. Infusions of saline (0.4 ml h(-1)) or hUII (30, 300 and 3,000 pmol kg(-1) h(-1)) were given i.v. for 6 h, and the effects of pretreatment with indomethacin (5 mg kg(-1) h(-1)), N(G)-nitro-L arginine methyl ester (L-NAME, 3 mg kg(-1) h(-1)) or propranolol (1 mg kg(-1); 0.5 mg kg(-1) h(-1)) on responses to hUII (300 pmol kg(-1) h(-1) for 6 h) were assessed. Cellular localisation of UT receptor-like immunoreactivity was determined in relevant tissues. hUII caused dose-dependent tachycardia and hindquarters vasodilatation, accompanied by a slowly developing rise in blood pressure. Haemodynamic effects of hUII were attenuated by propranolol or L-NAME and abolished by indomethacin. UT receptor-like immunoreactivity was detected in skeletal and vascular smooth muscle. The findings indicate that in conscious rats, infusions of hUII cause vasodilatation, which, of the vascular beds monitored, is selective for the hindquarters and dependent on cyclooxygenase products and nitric oxide. The pressor effect of hUII under these conditions is likely to be due to an increase in cardiac output, possibly due to a positive inotropic effect. UT receptor-like immunoreactivity present in skeletal muscle is consistent with the haemodynamic pattern. PMID- 16314854 TI - Long-term bradycardia caused by atrioventricular block can remodel the canine heart to detect the histamine H1 blocker terfenadine-induced torsades de pointes arrhythmias. AB - Although a second-generation histamine H(1) blocker terfenadine induced torsades de pointes (TdP) arrhythmias in patients via the blockade of a rapid component of delayed rectifier K(+) current (I(Kr)), such action of terfenadine has not been detected in previous animal models. We analysed the potential of the canine persistent atrioventricular block heart, a new in vivo proarrhythmia model, to detect a torsadogenic effect of terfenadine of an oral dose of 3 or 30 mg kg(-1). The doses can provide therapeutic to supra-therapeutic plasma concentrations as an anti-histamine. In 2 weeks of bradycardiac heart model, there were no significant changes in any of the electrocardiogram parameters after the administration of both doses of terfenadine. In 4-6 weeks of bradycardiac heart model, the low dose of terfenadine hardly affected any of the electrocardiogram parameters except that it induced TdP in one out of six animals. The high dose significantly decreased the atrial rate and ventricular rate, prolonged the QT interval, and induced TdP in five out of six animals. Moreover, temporal variability of repolarization increased after the high-dose administration. These results suggest that long-term bradycardia caused by atrioventricular block can remodel the canine heart to detect terfenadine-induced TdP. PMID- 16314855 TI - Gender differences in the slow delayed (IKs) but not in inward (IK1) rectifier K+ currents of canine Purkinje fibre cardiac action potential: key roles for IKs, beta-adrenoceptor stimulation, pacing rate and gender. AB - As the beagle dog is a commonly used preclinical species to test the effects of new drugs on cardiac repolarisation and Purkinje fibres have become an established in vitro preparation to assess the effects of these new drugs on action potential duration (APD), the main aim of this study was therefore to evaluate the relative contribution of the inward (I(K1)) and slow delayed (I(Ks)) rectifier cardiac K(+) currents to action potential repolarisation in beagle Purkinje fibres under three different experimental conditions: (i) selective block of I(K1) with BaCl(2), (ii) selective block of I(Ks) with (-) chromanol 293B under basal conditions and (iii) selective block of I(Ks) during beta adrenoceptor stimulation. Furthermore, the dependence of this contribution on gender and pacing rate was investigated. Microelectrode techniques were employed to measure APD in Purkinje fibres from adult female and male dogs. At stimulation rates of 3.33, 1.0 and 0.2 Hz, the degree of prolongation of APD evoked by BaCl(2) (10 microM) was comparable in fibres from female and male dogs. At the same stimulation rates, 10 microM (-) chromanol 293B did not change the APD in fibres from female and male dogs. During beta-adrenoceptor stimulation with 0.1 microM isoproterenol, an APD prolonging effect of (-) chromanol 293B was detected. In the presence of isoproterenol, action potentials in fibres from male dogs get shorter when changing the stimulation rate from 1.0 to 0.2 Hz, while the opposite is seen in fibres from female dogs. This alteration was completely reversed by (-) chromanol 293B. In conclusion, our findings confirm that beta adrenoceptor stimulation is one condition where there may be an increased role of I(Ks) in action potential repolarisation. Gender differences in the autonomic modulation of I(Ks) could be a contributing factor to the reported increased susceptibility of female hearts to arrhythmias. PMID- 16314856 TI - Neuroprotection associated with alternative splicing of NMDA receptors in rat cortical neurons. AB - Exposure of cultured cortical neurons to elevated extracellular K(+) concentrations (25 mM) induces membrane depolarization and an increase in action potential firing. Long-term high K(+) treatment was associated with an increased neuronal cell death. In surviving neurons, multiple changes occurred in the proportion of individual NMDA receptor subunit 1 (NR1) splice variant mRNA expression, whereas the overall expression of NR1, NR2A and NR2B transcripts remained unaffected. The high K(+)-induced changes in NR1 splice variant expression were virtually abolished upon a concurrent administration of tetrodotoxin (TTX; 3 microM). In voltage-clamp recordings performed on neurons resistant to high K(+) treatment, inward currents induced by NMDA (1-1,000 microM) were reduced. In K(+)-resistant cells, the activity of calpain but not of caspase-3 was diminished compared with controls kept in regular medium. NR function as well as calpain activity was not affected in cultures concomitantly treated with high K(+) and either TTX or a NR antagonist (CGS19755 (selfotel) or memantine). In conclusion, the present data indicate adaptive changes in NR1 splice variant expression and a decrease in NR function upon a sustained increase in neurotransmission. Accordingly, alternative splicing could be an endogenous mechanism to counteract cellular damage due to overactivation of excitatory NRs and may be associated with an impairment of necrotic mechanisms. PMID- 16314858 TI - Power tools for human genetics. PMID- 16314859 TI - Gaining insight into PTPN22 and autoimmunity. PMID- 16314860 TI - SILencing misbehaving proteins. PMID- 16314861 TI - Aging (not so) gracefully. PMID- 16314866 TI - C. elegans cell cycles: invariance and stem cell divisions. AB - The adult Caenorhabditis elegans nematode, a small roundworm, has a precisely defined number of somatic cells that create organs that are also found in larger animals, including intestine, muscles, skin, an excretory system and a primitive brain. Every cell has a defined role in this sophisticated, but tiny animal. Therefore, stringent control of the cell cycle is required to produce the almost invariant cell lineage that generates the C. elegans somatic body plan. The proliferation of germ cells is regulated differently, and occurs within a stem cell niche. PMID- 16314867 TI - The cell biology of neurogenesis. AB - During the development of the mammalian central nervous system, neural stem cells and their derivative progenitor cells generate neurons by asymmetric and symmetric divisions. The proliferation versus differentiation of these cells and the type of division are closely linked to their epithelial characteristics, notably, their apical-basal polarity and cell-cycle length. Here, we discuss how these features change during development from neuroepithelial to radial glial cells, and how this transition affects cell fate and neurogenesis. PMID- 16314868 TI - Semaphorins command cells to move. AB - Semaphorins are secreted or transmembrane proteins that regulate cell motility and attachment in axon guidance, vascular growth, immune cell regulation and tumour progression. The main receptors for semaphorins are plexins, which have established roles in regulating Rho-family GTPases. Recent work shows that plexins can also influence R-Ras, which, in turn, can regulate integrins. Such regulation is probably a common feature of semaphorin signalling and contributes substantially to our understanding of semaphorin biology. PMID- 16314869 TI - Tetraspanin functions and associated microdomains. AB - Cell-surface proteins of the tetraspanin family are small, and often hidden by a canopy of tall glycoprotein neighbours in the cell membrane. Consequently, tetraspanins have been understudied and underappreciated, despite their presence on nearly all cell and tissue types. Important new genetic evidence has now emerged, and is bolstered by new insights into the cell biology, signalling and biochemistry of tetraspanins. These new findings provide a framework for better understanding of these mysterious molecules in the regulation of cellular processes, from signalling to motility. PMID- 16314870 TI - Serine racemase binds to PICK1: potential relevance to schizophrenia. AB - Accumulating evidence from both genetic and clinico-pharmacological studies suggests that D-serine, an endogenous coagonist to the NMDA subtype glutamate receptor, may be implicated in schizophrenia (SZ). Although an association of genes for D-serine degradation, such as D-amino acid oxidase and G72, has been reported, a role for D-serine in SZ has been unclear. In this study, we identify and characterize protein interacting with C-kinase (PICK1) as a protein interactor of the D-serine synthesizing enzyme, serine racemase (SR). The binding of endogenous PICK1 and SR requires the PDZ domain of PICK1. The gene coding for PICK1 is located at chromosome 22q13, a region frequently linked to SZ. In a case control association study using well-characterized Japanese subjects, we observe an association of the PICK1 gene with SZ, which is more prominent in disorganized SZ. Our findings implicating PICK1 as a susceptibility gene for SZ are consistent with a role for D-serine in the disease. PMID- 16314871 TI - Why do young women smoke? I. Direct and interactive effects of environment, psychological characteristics and nicotinic cholinergic receptor genes. AB - Despite the health hazards, cigarette smoking is disproportionately frequent among young women. A significant contribution of genetic factors to smoking phenotypes is well established. Efforts to identify susceptibility genes do not generally take into account possible interaction with environment, life experience and psychological characteristics. We recruited 501 female Israeli students aged 20-30 years, obtained comprehensive background data and details of cigarette smoking and administered a battery of psychological instruments. Smoking initiators (n=242) were divided into subgroups with high (n=127) and low (n=115) levels of nicotine dependence based on their scores on the Fagerstrom Tolerance Questionnaire and genotyped with noninitiators (n=142) for single nucleotide polymorphisms (SNPs) in 11 nicotinic cholinergic receptor genes. We found nominally significant (P<0.05) allelic and genotypic association with smoking initiation of SNP rs2072660 and multilocus haplotypes (P<0.007-0.05) in CHRNB2 and nominal (P<0.05) allelic or genotypic association of SNPs in CHRNA7 (rs1909884), CHRNA9 (rs4861065) and CHRNB3 (rs9298629) with nicotine dependence. Employing logistic regression and controlling for known risk factors, the best fitting model for smoking initiation encompassed a 5 SNP haplotype in CHRNB2, neuroticism and novelty seeking (P=5.9 x 10(-14), Nagelkerke r(2)=0.30). For severity of nicotine dependence, two SNPs in CHRNA7 (rs1909884 and rs883473), one SNP in CHRNA5 (rs680244) and the interaction of a SNP in CHRNA7 (rs2337980) with neuroticism, were included in the model (P=2.24 x 10(-7), Nagelkerke r(2)=0.40). These findings indicate that background factors, psychological characteristics and genetic variation in nicotinic cholinergic receptors contribute independently or interactively to smoking initiation and to severity of nicotine dependence in young women. PMID- 16314872 TI - Association of galanin haplotypes with alcoholism and anxiety in two ethnically distinct populations. AB - The neuropeptide galanin (GAL) is widely expressed in the central nervous system. Animal studies have implicated GAL in alcohol abuse and anxiety: chronic ethanol intake increases hypothalamic GAL mRNA; high levels of stress increase GAL release in the central amygdala. The coding sequence of the galanin gene, GAL, is highly conserved and a functional polymorphism has not yet been found. The aim of our study was, for the first time, to identify GAL haplotypes and investigate associations with alcoholism and anxiety. Seven single-nucleotide polymorphisms (SNPs) spanning GAL were genotyped in 65 controls from five populations: US and Finnish Caucasians, African Americans, Plains and Southwestern Indians. A single haplotype block with little evidence of historical recombination was observed for each population. Four tag SNPs were then genotyped in DSM-III-R lifetime alcoholics and nonalcoholics from two population isolates: 514 Finnish Caucasian men and 331 Plains Indian men and women. Tridimensional Personality Questionnaire harm avoidance (HA) scores, a dimensional measure of anxiety, were obtained. There was a haplotype association with alcoholism in both the Finnish (P=0.001) and Plains Indian (P=0.004) men. The SNPs were also significantly associated. Alcoholics were divided into high and low HA groups (>or= and 3 mg/l. No differences were observed between men and women even after adjustment for age and body mass index (BMI). After patients with CRP values above the 97.5th percentile (n = 31) had been excluded, CRP concentrations increased significantly with increasing levels of cardiovascular risk factors in both men and women. Men and women with metabolic syndrome showed significantly higher levels of CRP than their counterparts, even after adjustment for BMI and age. In a multiple regression analysis, BMI, triglycerides and fasting glucose were independent predictors of CRP in women and together explained 42% of its variance. In men, CRP was independently and positively associated to waist circumference, smoking, diastolic blood pressure, uric acid and triglycerides, and negatively associated to HDL-cholesterol. Altogether these variables explained 51% of its variability. CONCLUSIONS: The present study describes, for the first time, CRP concentrations in a sample that is representative of a Spanish Mediterranean community. CRP distribution and correlates are very similar to those reported previously in spite of the different lifestyle and nutritional habits, and the lower rates of cardiovascular diseases in our population. PMID- 16314876 TI - Cardiac parasympathetic regulation in obese women with binge eating disorder. AB - OBJECTIVE: Obese individuals with a binge eating disorder (BED) differ from obese non-binge eaters (NBED) with respect to (a) eating behaviour, (b) psychiatric comorbidity and (c) level of psychosocial distress. The aim of the study was to explore whether these three factors have an influence on cardiac parasympathetic function, that is independent of obesity: as alterations in cardiac parasympathetic function may have a role in the higher cardiovascular mortality that is present in obese individuals. METHODS: In total, 38 obese women (BMI>30 kg/m(2)), with a BED and 34 age and BMI matched healthy controls (NBED) completed a laboratory stress protocol that incorporated a baseline resting period, Head-up Tilt Testing (HUT) and two challenging mental tasks. Heart rate and blood pressure were measured continuously during the protocol. Parasympathetic cardiac regulation was assessed as the high frequency component of heart rate variability (HRV-HF). RESULTS: Mental challenge led to an augmented reduction of HRV-HF in obese binge eaters, which was linked to the binge eating frequency and hunger perception, but not to psychiatric comorbidity. During baseline conditions and HUT, no significant differences in parasympathetic measures were observed between the two subject groups. CONCLUSION: Subjects with a BED showed greater reduction in parasympathetic cardiac control (HRV-HF) during mental stress, suggesting higher stress vulnerability in women with a BED. Longitudinal investigations are necessary to evaluate whether this is associated with an increased cardiovascular mortality. PMID- 16314877 TI - Effects of peanut oil consumption on appetite and food choice. AB - OBJECTIVE: Peanut consumption may improve lipid profiles without promoting weight gain. Both properties have been attributed to their high-unsaturated fat content. Mono and polyunsaturated fatty acids reportedly hold stronger satiety value than saturated fats and may help appetite control. This study investigated the effects of chronic peanut oil consumption on appetite and food choice. RESEARCH METHODS AND PROCEDURES: A total of 129 healthy adults from three countries (Brazil, Ghana and US) were randomly assigned to one of four treatment arms: consumption of peanut oil, olive oil or safflower oil as 30% of individual resting energy expenditure (REE) for 8 weeks or no dietary intervention. Participants received no other dietary guidance. They completed appetite questionnaires eliciting information about hunger, fullness, desire to eat, and prospective consumption during all waking hours for 1 day at weeks 2 and 6 and for 1 or 3 days at weeks 0, 4 and 8. Diet records were completed at weeks 0, 4 and 8. RESULTS: No differences in appetitive ratings were observed over the 8-week trial. There were no significant treatment by time interactions. Total caloric intake was significantly higher at week 8 relative to baseline (F=10.08, P<0.05). The increases for each treatment were: peanut oil=197+/-114; olive oil=237+/-121; safflower oil=274+/-90; control=75+/-71. Free-feeding intake, an index of dietary compensation, was reduced significantly at weeks 4 and 8 compared to baseline (F=9.08, P<0.00). The declines (compensation scores) were (kcals): peanut oil= 208+/-105 (46%); olive oil=-235+/-105 (50%); safflower oil=-186+/-102 (44%). There were no significant differences across countries in appetite ratings. DISCUSSION: A prior intervention with whole peanuts reported a dietary compensation score of 66% over 8 weeks, this compares to a 46% compensation score observed with peanut oil. Our data suggests that the lipid fraction in peanuts elicits a weak effect on satiety. PMID- 16314878 TI - The effects of changing exercise levels on weight and age-related weight gain. AB - OBJECTIVE: To determine prospectively whether physical activity can prevent age related weight gain and whether changing levels of activity affect body weight. DESIGN/SUBJECTS: The study consisted of 8,080 male and 4,871 female runners who completed two questionnaires an average (+/-standard deviation (s.d.)) of 3.20+/ 2.30 and 2.59+/-2.17 years apart, respectively, as part of the National Runners' Health Study. RESULTS: Changes in running distance were inversely related to changes in men's and women's body mass indices (BMIs) (slope+/-standard error (s.e.): -0.015+/-0.001 and -0.009+/-0.001 kg/m(2) per Deltakm/week, respectively), waist circumferences (-0.030+/-0.002 and -0.022+/-0.005 cm per Deltakm/week, respectively) and percent changes in body weight (-0.062+/-0.003 and -0.041+/-0.003% per Deltakm/week, respectively, all P<0.0001). The regression slopes were significantly steeper (more negative) in men than women for DeltaBMI and Delta%body weight (P<0.0001). A longer history of running diminished the impact of changing running distance on men's weights. When adjusted for Deltakm/week, years of aging in men and years of aging in women were associated with increases of 0.066+/-0.005 and 0.056+/-0.006 kg/m(2) in BMI, respectively, increases of 0.294+/-0.019 and 0.279+/-0.028% in Delta%body weight, respectively, and increases of 0.203+/-0.016 and 0.271+/-0.033 cm in waist circumference, respectively (all P<0.0001). These regression slopes suggest that vigorous exercise may need to increase 4.4 km/week annually in men and 6.2 km/week annually in women to compensate for the expected gain in weight associated with aging (2.7 and 3.9 km/week annually when correct for the attenuation due to measurement error). CONCLUSIONS: Age-related weight gain occurs even among the most active individuals when exercise is constant. Theoretically, vigorous exercise must increase significantly with age to compensate for the expected gain in weight associated with aging. PMID- 16314879 TI - The impact of the covert manipulation of macronutrient intake on energy intake and the variability in daily food intake in nonobese men. AB - OBJECTIVE: To investigate the effect of macronutrient composition on ad libitum food intake in nonobese men. DESIGN: Balanced, incomplete-block, crossover study where subjects received two of three treatments. Macronutrient composition was manipulated by providing 2.1 MJ/day high-carbohydrate (CHO), high-fat (FAT), and/or high-protein (PRO) drinks every day over the course of two, 8-week periods. SUBJECTS: In all, 12 healthy normal weight men (age: 39+/-9 years, BMI: 24.1+/-1.4 kg/m2). MEASUREMENTS: Ad libitum food intake was measured continuously for 16 weeks at the Beltsville Human Nutrition Research Center (BHNRC). Body composition (DEXA) and body weight were also measured. RESULTS: Average energy intake (EI) during weeks 1 and 2 was lower for CHO than FAT (P<0.05), but this effect disappeared by week 3. EI during CHO increased by 11% from week 1 to 8 through the increased selection of carbohydrate and protein-containing foods, but not fat foods. Food intake was variable, both between and within subjects, but was not related to macronutrient composition. CONCLUSION: EI appears to be influenced by macronutrient composition in the short-term when diets are modified, but the effect dissipates in a few weeks if the diet is maintained. These data suggest the presence of macronutrient-specific regulatory mechanisms in the body, but do not support the notion that a high intake of any of the three macronutrients suppresses EI over a prolonged period of time. The high variability in food intake does not appear to be related to macronutrient composition. PMID- 16314880 TI - (AAT)n repeat in the cannabinoid receptor gene (CNR1): association with cocaine addiction in an African-Caribbean population. AB - Owing to their agonist action on dopaminergic systems, cannabinoids may play a major role in substance dependency and schizophrenia. We examined the (AAT)n triplet repeat polymorphism nearby the CNR1 gene, which encodes human cannabinoid (CB1) receptor, in a male Afro-Caribbean population. The allelic and genotypic distributions were significantly different in non-schizophrenic cocaine dependents (n = 97), schizophrenic cocaine dependents (n = 45) and matched controls (n = 88) (P < 10(-4)). The frequency of the (AAT)12 repeat allele was increased in non-schizophrenic cocaine dependents and schizophrenic cocaine dependents vs controls (25.3 and 26.7 vs 5.7%) (P < 10(-4)). Our results support that the (AAT)n polymorphism nearby the CNR1 gene could be associated with predisposition to cocaine dependency. PMID- 16314881 TI - Comparative genomics analysis of human sequence variation in the UGT1A gene cluster. AB - Common polymorphisms within the human UGT1A gene locus are associated with irinotecan and tranilast toxicity. To uncover additional functional variation across this gene cluster, cross-species sequence comparisons were performed. Evolutionarily conserved segments (a total of 47.1 kb) were re-sequenced in 24 African-American, 24 European-American, and 24 Asian individuals, and 381 segregating sites (including 123 singletons) were identified. Highly conserved coding sites were less likely to be polymorphic than diverged sites (P<0.0001) but this pattern was not observed at non-coding sites (P=0.1025). Among coding variants, the distribution of those computationally predicted to affect function was skewed toward low frequencies. Some alleles occurred at similar frequencies in each population; others had wide disparities. Although strong linkage disequilibrium was detected among the hepatically expressed genes, the degree of linkage disequilibrium varied among populations. These results suggest that rare functional gene variants and inter-population variability must be considered in the interpretation of association studies between UGT1A and drug metabolism/toxicity phenotypes. PMID- 16314882 TI - Sequence diversity and haplotype structure at the human CYP3A cluster. AB - The four members of the human CYP3A subfamily play important roles in the clearance of xenobiotics, hormones, and environmental compounds. Many SNPs at the CYP3A locus have been characterized, with several showing large allele frequency differences across populations. In addition to the effects of CYP3A SNPs on drug metabolism, recent studies have highlighted the potential for CYP3A variation in susceptibility to several common phenotypes, including hypertension and cancer. We previously showed that the CYP3A4 and CYP3A5 genes have a strong haplotype structure at varying frequencies across ethnic groups. Here, we extend our re sequencing survey to the remaining CYP3A genes in the same cluster, CYP3A7 and CYP3A43. Our study identified a large number of SNPs in coding and conserved noncoding sequences, several of which are common. The combined data set allows us to investigate patterns of sequence variation and linkage disequilibrium at the entire CYP3A locus for use in future association studies. PMID- 16314883 TI - Ionotropic glutamate receptor gene GRIK3 SER310ALA functional polymorphism is related to delirium tremens in alcoholics. AB - Upregulation of glutamatergic neurotransmission resulting from chronic ethanol intoxication may cause a hyperexcitable state during alcohol withdrawal, which may lead to seizures and delirium tremens. The aim of our study was to evaluate the association between a history of alcohol withdrawal-induced seizures and delirium tremens, and a functional polymorphism (Ser310Ala) of the GRIK3 gene coding for the glutamatergic kainate receptor subunit GlurR7 in a sample of well characterized alcoholics compared to controls. In total, 233 patients meeting DSM IV alcohol dependence criteria and 309 controls, all of German descent, were investigated. GRIK3 functional polymorphism was determined using PCR (polymerase chain reaction) of lymphocyte DNA. History of alcohol withdrawal-induced delirium tremens and seizures were obtained using the SSAGA (Semi-Structured Assessment for the Genetics of Alcoholism). Data were cross-checked with in-patients' clinical files. While a significant relationship between history of delirium tremens and the Ser310 allele was detected, no significant results were obtained for alcohol withdrawal-related seizures. Although this result is suggestive for a significant role of this polymorphism in the pathogenesis of delirium tremens in alcohol-dependent individuals, further investigation and confirmation are warranted. PMID- 16314884 TI - Pharmacologic analysis of non-synonymous coding h5-HT2A SNPs reveals alterations in atypical antipsychotic and agonist efficacies. AB - The 5-HT(2A)-serotonin receptor is a major molecular target for most atypical antipsychotic drugs as well as most hallucinogens, which can exacerbate psychotic symptoms. In this study, we examined whether random sequence variations in the gene (single nucleotide polymorphisms, SNPs) encoding the 5-HT(2A)-serotonin receptor could explain inter-individual variability in atypical antipsychotic and agonist drug response. We examined the in vitro pharmacology of four non synonymous SNPs, which give rise to T25N, I197V, A447V, and H452Y variant 5 HT(2A)-serotonin receptors. Our data indicate that these non-synonymous SNPs exert statistically significant, although modest, effects on the affinity and functional effects of several currently approved atypical antipsychotics (aripiprazole, clozapine, olanzapine, quetiapine, risperidone, and ziprasidone). Also, the 5-HT(2A) receptor SNPs slightly altered the potency and relative efficacy of a small number of selected agonists (2,5-dimethoxy-4-iodoamphetamine, tryptamine, 5-hydroxytryptamine, m-chlorophenylpiperazine, and 5-methoxy-N, N dimethyltryptamine). In all, our results show that the in vitro pharmacological effects of the SNPs are drug specific. PMID- 16314885 TI - The current clinical practice of pharmacogenetic testing in Europe: TPMT and HER2 as case studies. PMID- 16314886 TI - Interactions between five candidate genes and antihypertensive drug therapy on blood pressure. AB - Despite the availability of effective antihypertensive drugs, there is a large variation in response to these drugs. This study investigates whether polymorphisms in the angiotensin converting enzyme (I/D), angiotensinogen (M235T), alpha-adducin (G460W), angiotensin II type 1 receptor (1166A/C), or G protein beta(3)-subunit (825C/T) gene modify the mean difference in blood pressure levels among diuretics, beta-blockers, or ACE-inhibitors users. Data were used from the Doetinchem Cohort Study, and blood pressure data were collected from GPs (1987-1997). A marginal generalized linear model (GEE) was used to assess the gene-drug interaction on the mean difference in systolic/diastolic blood pressure. In total, 625 hypertensive individuals were included with a total of 5262 measurements of blood pressure. Only the interaction between diuretic use and the GNB3 825C/T polymorphism was significant (C allele versus TT systolic blood pressure (SBP): 4.33 mmHg [95% CI: 0.14 8.54]). Thus, the mean SBP level among diuretic users may be modified by the GNB3 825C/T polymorphism. PMID- 16314887 TI - The expanding role of BDNF: a therapeutic target for Alzheimer's disease? AB - Finding an effective treatment for chronic neurodegenerative disorders still represents an unmet goal. There is considerable evidence that such disorders represent a combination of genetic determinants and failure of neuroprotective mechanisms sparking a wider degree of interest in shedding light on the cellular changes responsible for these devastating disorders. Because of their role in survival or differentiation of developing neurons, as well as the recent discovery of their importance in regulating synaptic plasticity during adulthood, neurotrophic factors have been suggested as essential contributors of the etiology of neurodegenerative disorders. Alzheimer's disease (AD) is a complex, chronic, devastating disease that affects a high percentage of the population over 65 years of age. This review will focus on different pharmacological interventions that are currently in use or drugs under development, narrowing the therapeutic agents to those that interfere with the expression of the trophic factor brain-derived neurotrophic factor (BDNF), a molecule playing a pivotal role in synaptic plasticity and cognition. From these findings, it appears clear that BDNF is implicated in the mechanism of action of drugs that improve cognitive deficits in animal models of AD and in AD patients. PMID- 16314888 TI - Haplotype structures of the UGT1A gene complex in a Japanese population. AB - Genetic polymorphisms of UDP-glucuronosyltransferases (UGTs) are involved in individual and ethnic differences in drug metabolism. To reveal co-occurrence of the UGT1A polymorphisms, we first analyzed haplotype structures of the entire UGT1A gene complex using the polymorphisms from 196 Japanese subjects. Based on strong linkage disequilibrium between UGT1A8 and 1A10, among 1A9, 1A7, and 1A6, and between 1A3 and 1A1, the complex was divided into five blocks, Block 8/10, Block 9/6, Block 4, Block 3/1, and Block C, and the haplotypes for each block were subsequently determined/inferred. Second, using pyrosequencing or direct sequencing, additional 105 subjects were genotyped for 41 functionally tagged polymorphisms. The data from 301 subjects confirmed the robustness of block partitioning, but several linkages among the haplotypes with functional changes were found across the blocks. Thus, important haplotypes and their linkages were identified among the UGT1A gene blocks (and segments), which should be considered in pharmacogenetic studies. PMID- 16314889 TI - Salvage prostate cryoablation after primary interstitial brachytherapy failure: a feasible approach. AB - Salvage treatment for recurrent prostate cancer remains a very difficult and challenging field in urologic oncology. The introduction of minimally invasive surgical procedures such a targeted cryoablation brings some hope with its feasibility and efficacy to become a potentially curable treatment. We present the case of a 75-year-old male with prostate cancer treated primarily by brachytherapy, who developed late locally recurrent disease that was successfully treated with targeted salvage cryoablation. PMID- 16314890 TI - Frequency of impalpable prostate adenocarcinoma and precancerous conditions in Greek male population: an autopsy study. AB - A series of 212 prostate specimens of men dead between August 2002-August 2004, have been sectioned in consecutive autopsies and subjected to whole mount analysis in purpose to determine the epidemiology of impalpable prostate cancer in Greece. Impalpable prostate carcinomas were found in 40 cases (18.8%) most in the peripheral region. In all, 29 of 40 impalpable cancers (70.7%) had volume less than 1 cm3. Most of impalpable carcinomas were of favorable (Gleason score 2 4) or intermediate (Gleason score 5 and 6) histological type (55 and 27.7%, respectively) while only five (12.5%) were undifferentiated (Gleason score 7 and 8). In all, 24 (60%) of the 40 impalpable carcinomas were multifocal and consisted of two or more foci, most of small size (<0.5 cm3). Most of the impalpable cancers found in this autopsy study were potentially insignificant tumors (relatively low volume, favorable or intermediate histological pattern and absence of invasiveness). Prostate intraepithelial neoplasia (PIN) coexisted with impalpable carcinomas, in almost half of the cancer-positive specimens. There was a positive correlation between PIN foci and coexistent cancer foci in most of the cases. Frequency rate and pathological features of both entities show significant variations in medical literature. Since the incidence of clinical prostate cancer in Greece is relatively low, and according to our autopsy findings, it is plausible that the frequency of clinical prostate cancer in a certain population could be related to the prevalent model of impalpable cancer as well as to the frequency and extend of the precancerous lesions. PMID- 16314891 TI - Novel antiproliferative flavonoids induce cell cycle arrest in human prostate cancer cell lines. AB - Epidemiologic studies have demonstrated an inverse association between flavonoid intake and prostate cancer (PCa) risk. The East Asian diet is very high in flavonoids and, correspondingly, men in China and Japan have the lowest incidence of PCa worldwide. There are thousands of different naturally occurring and synthetic flavonoids. However, only a few have been studied in PCa. Our aim was to identify novel flavonoids with antiproliferative effect in PCa cell lines, as well as determine their effects on cell cycle. We have screened a representative subgroup of 26 flavonoids for antiproliferative effect on the human PCa (LNCaP and PC3), breast cancer (MCF-7), and normal prostate stromal cell lines (PrSC). Using a fluorescence-based cell proliferation assay (Cyquant), we have identified five flavonoids, including the novel compounds 2,2'-dihydroxychalcone and fisetin, with antiproliferative and cell cycle arresting properties in human PCa in vitro. Most of the flavonoids tested exerted antiproliferative effect at lower doses in the PCa cell lines compared to the non-PCa cells. Flow cytometry was used as a means to determine the effects on cell cycle. PC3 cells were arrested in G2/M phase by flavonoids. LNCaP cells demonstrated different cell cycle profiles. Further studies are warranted to determine the molecular mechanism of action of 2,2'-DHC and fisetin in PCa, and to establish their effectiveness in vivo. PMID- 16314892 TI - PSA testing: an evolving relationship with prostate cancer screening. AB - PSA testing has made prostate cancer screening a reality for men in many parts of the world, but its benefit for men's health continues to be debated. In men exposed to PSA testing, there has been a well-documented change in the presentation of prostate cancer with a shift towards earlier pathological stage, not without justifiable concern about over-diagnosis by prostate biopsy. Increasingly, men now diagnosed with early stage cancer have previous PSA exposure and are selected for biopsy based on PSA change in relation to cutoff values. Some recent observations suggest that PSA may no longer be an effective marker for early stage tumours, with PSA elevation failing to discriminate tumour specific characteristics from benign gland enlargement. Traditionally, variation in pathological stage of clinically localised prostate cancer at diagnosis has related to clinical stage, PSA and biopsy Gleason grade, but with distinctions based upon these three assessments declining and an increasing proportion of organ-confined tumours at presentation, new methods of cancer detection and prognostic assessment are now required. Molecular technologies hold great promise in this respect, and in the future biomarker signatures are likely to overshadow total PSA for guiding early diagnosis and prognostic assessment. While arguments about prostate screening will continue, owing not least to its feasibility, future debate is likely to focus increasingly on technological advances and molecular profiling of these notoriously heterogeneous tumours. PMID- 16314893 TI - [Hospitalization of patients with eating disorders: an eight years review]. AB - Eating disorders (Eds) are pathologies with a multifactorial etiopathogenesis. Their treatment requires an interdisciplinary team. The aim of this paper is to discuss in patient treatment (n=80) of patients with eating disorders during eight years. This treatment was in charge of the Eating Disorders Team of the Hospital Italiano of Buenos Aires. The approach to critical stages of eating disorders envolves the clinical or psychiatric admission of the patients as the principal means to ward off patients from death or the appearance of cronifications. 77,5% of the patients were admitted to the psychiatric ward, 5% to a clinical ward and 17,5% to a clinical followed by a psychiatric ward. The average B.M.I. in admission was 16,83. The average time in psychiatry was 35 days. During inpatient treatment 21,25% required a nasogastric tube and 82% received psychopharmacological treatment. The mortality rate was 1,25%. During the ambulatory treatment that followed hospitalizations, 32,5% were readmitted to the psychiatric ward and there were 10% of dropouts. Hospitalization of these patients looks after the achievement of healthier eating habits and the establishment of a therapeutic alliance which allows the follow up of an ambulatory treatment. PMID- 16314894 TI - [Post-traumatic stress disorder and substance use disorder: treatment intervention]. AB - There is an increasing interest to find efficacious treatments for patients with post-traumatic stress disorder (PTSD) and comorbid substance use disorder (SUD). Therefore, in this paper the studies published on the psychological and pharmacological treatment for the management of comorbid PTSD and SUD will be discussed, as these are highly prevalent and shown in part I. The few studies published to date suggest that patients benefit most form an integrated treatment approach for the comorbid disorders. Psychological treatments based on cognitive behavioural strategies appear to be useful for patients with PTSD and comorbid SUD disorder. Medications such as SSRIs and particularly sertraline that efficacious for PTSD, seem to be as well for this dual disorder. In addition, pharmacological options, among them quetiapine or bupropion have shown promising results in preliminary studies. PMID- 16314895 TI - [Visual art, creativity and dementia]. AB - BACKGROUND: Visual art is an expression of neurological function and how it organizes and interprets perception. The art is predominantly in the right hemisphere, in contrast, the left side, have inhibitory effects on artistic expression. In normal subjects, inhibitory and excitatory mechanisms could interact in a complex harmony, reflecting a paradoxical functional facilitation. Brain diseases such as dementia could change this harmony and then, alter the artistic abilities. OBJECTIVE: Evaluate the art expression in the degenerative diseases. PATIENTS AND METHODS: Artistic abilities of 3 painters with degenerative diseases were assessment. RESULTS: Patient 1: A 83 - year old right handed female, diagnosis: Alzheimer's disease. Artistic description: low productivity, simplified versions of earlier and alteration of the visuospatial organization. Patient 2: A 78-year-old right handed female, diagnosis: Primary Progressive Aphasia (PPA); Artistic description: oversimplified drawings which maintaining overall spatial organization, without impair artistic skills. Patient 3: A 68 year-old right handed woman, diagnosis: Fronto-Temporal Dementia (FTD). Artistic description: Increased artistic activity, originality, freedom, utilization of intense colours with perseverative and repetitive copying of similar paintings of her own work. CONCLUSIONS: Visual art in Alzheimer's disease is a consequence of visuospatial and constructive disabilities. In contrast, the conservation of this cognitive functions and left asymmetrical involved, in FTD and PPA respectively, suggest artistic preservation, independently of the language injury. The disproportionate functional prevalence of the right over the left could lead to a release of novelty - seeking in art and can contribute to emergent creativity. These observations suggest an organization for art in the brain and proposed bases for further investigations in dementias. PMID- 16314896 TI - [Notes for a discussion on the notion of resistance in psychopharmacology]. AB - The present work's purpose is to open a discussion on the notion of Resistance in Psychopharmacology. Despite the fact that the phenomenon is observable in psychiatric practice, its meaning and consequences for medical practice are not sufficiently established, so that a critical reformulation is required for the sake of greater conceptual clarity in this area. PMID- 16314897 TI - [Pharmacokinetic mechanisms underlying resistance in psychopharmacological treatment. The role of P-glycoprotein]. AB - In the last years efflux transporters, as for example P-glycoprotein, have been shown to play an important role in the regulation of the uptake of drugs in the central nervous system. These transporters are expressed constitutively in the brain capillary endothelial cells which form the brain-blood barrier, but their expression or activity could be inhibited or induced by other compounds or could be modified under pathological conditions. Some antipsychotics (amisulpride, chlorpromazine, fluphenazine, haloperidol, olanzapine, pimozide, prometazine, quetiapine, risperidone, trifluoperazine), antidepressants (amitriptiline, doxepine, nortriptiline, venlafaxine), and antiepileptics (felbamate, gabapentin, lamotrigine, phenobarbital and topiramate) are P-glycoprotein substrates. Interactions that could take place at the P-glycoprotein level may explain some cases of resistance in polimedicated patients. Other factors, yet unknown, could induce the expression of this transporter and therefore decrease the uptake of psychotropic drugs in the central nervous system, affecting their efficacy. PMID- 16314898 TI - [Pharmacological guidelines for the treatment of refractory patients with obsessive-compulsive disorder]. AB - OCD is a chronic and disabling anxiety disorder, it has a chronic course and few patients achieve true remission. OCD is clearly associated with a significant functional disability. Treatment with selective serotonin reuptake inhibitors (SSRIs) is effective in 40 % to 60 % of patients with OCD, but that leaves a large percentage of patients who are non responsive to treatment. This article will discuss guidelines for the use of SSRIs and novel approaches for managing treatment-refractory patients. PMID- 16314899 TI - [Treatment resistant schizophrenia: therapeutic approaches]. AB - An important percentage of schizophrenic patients does not respond or responds partially to treatment. The concept of schizophrenia resistance has changed over the years, moving from a dichotomous model into a continuous one. Although clozapine is the only antipsychotic with proved efficacy for these patients, beneficial effects have been observed with other drugs. Other antipsychotics, their combination, anticonvulsants, lithium, glycine site agonists, antidepressants and benzodiazepines are also used in the treatment of refractory patients with diverse results. In this article, evidence of the efficacy of these drugs and other types of treatment on schizophrenic- resistant patients is reviewed, and a psychopharmacological treatment strategy is proposed. PMID- 16314900 TI - [Resistance in the treatment of bipolar disorder: conceptual issues and therapeutic consequences]. AB - The concept of resistance in the treatment of bipolar disorders is not adequately formalized at present. This is due to the complexity of the evaluation of treatment effectiveness in a clinical scenario of heterogeneous development where huge differences between the degrees of recuperation achieved by different groups of patients can be observed. The use of an oversimplified concept of resistance to treatment favours the appearance of inadequate therapeutic strategies and the inclusion of treatment proposals devoid of evidence. On the other hand, a concept of resistance which does not take into account the clinical characteristics of these cases makes the understanding of results difficult for patients in treatment. Certain psychotherapies and pharmacological tools do at least share a level of consensus that grant their suggestion in cases of poor response to conventional therapeutics. PMID- 16314901 TI - [Pharmacological treatment of resistant depression]. AB - This article will review the evaluation and treatment of the resistant depressed patient. Although most depressives improve rapidly, a relatively small group is resistant to acute treatment. For resistant depression the first strategy must be thorough reassessment. This will include review of the diagnosis and the type of previous treatment. Defining adequacy of treatment is not always an easy task, and inadequate antidepressant trials are a relatively common phenomenon. Various treatment options include: augmentation, combination, and switching; the vast majority of available evidence on the efficacy of augmentation with lithium and thyroid supplementation indicate a remarkable degree of consistency in concluding that is effective; there is limited evidence, mostly provided by uncontrolled studies, supporting the efficacy of combining antidepressants; options of switching to a different monotherapy (to other antidepressant) are reviewed. Different strategies may be efficacious in the treatment of resistant depression; however, further randomized controlled trials with larger sample sizes are required to demonstrate the efficacy of options of treatment-resistant depression. PMID- 16314902 TI - [Extrapyramidal complications during treatment with chlorpromazine and reserpine: clinical and electromyographic study]. PMID- 16314903 TI - [A historical-epistemological perspective of the ontological problem of Positivism. Positivism and psychiatry]. AB - The present article reviews epistemological problems related to the philosophical and scientific current known as Positivism. The revision begins with the study of the historical period during which the Logical Positivism emerges (early in the decade of 1920); and finishes with the analysis of its influence on psychiatry and its relation with other epistemological currents in the same period. The article reviews the historical and epistemological outlook at te beginning of the XX th century, when the problem of language became ontologically outstanding. The question of language remains important in the present discussions. PMID- 16314904 TI - Aortic elastic properties and left ventricular diastolic dysfunction in patients with obstructive sleep apnea. AB - Although the responsible mechanisms are not yet fully known, obstructive sleep apnea is associated with an increased risk for cardiovascular disease and events. The aorta is not only a conduit delivering blood to the tissues but is also an important modulator of the entire cardiovascular system, its elastic properties also affecting left ventricular function and coronary blood flow. The aim of this study was to determine left ventricular diastolic function and aortic elastic properties in patients with obstructive sleep apnea syndrome. Fourteen male patients with obstructive sleep apnea and 14 age- and body mass index-matched healthy male controls took part in the study as a control group. All subjects underwent echocardiographic examination; left ventricular cavity dimension, standard and tissue Doppler parameters, and aortic diameter (3 cm above aortic valve) at systole and diastole were measured. While the aortic stiffness index in patients with obstructive sleep apnea was significantly higher than that of the control group (4.5 +/- 0.3 vs 2.1 +/- 0.1, P = 0.001), the aortic distensibility index was found to be lower in this group compared with controls (2.4 +/- 1.2 vs 3.9 +/- 1.5 cm2 dynes(-1) 10(-6), P = 0.009). Furthermore, peak velocity of myocardial systolic wave and peak velocities of myocardial diastolic waves in sleep apnea patients were lower than in controls. There was an association between aortic stiffness and the apnea hypopnea index (coefficient = 0.49, P = 0.002). We also found an inverse correlation between peak velocity of myocardial diastolic wave and aortic stiffness (coefficient = -0.43, P = 0.003), using multiple linear regression. Increased aortic stiffness that is associated with the severity of disease in patients with obstructive sleep apnea may lead to diastolic dysfunction of the left ventricle. PMID- 16314905 TI - Inverse correlation between soluble CD40 ligand and soluble CD40 is absent in patients with unstable angina. AB - The CD40/CD40 ligand (CD40L) system mediates inflammatory processes important in atherogenesis and plaque instability. The expression of CD40L on activated T cells was suppressed by soluble CD40 (sCD40) in vitro. However, the relationship between soluble CD40L (sCD40L) and sCD40 in unstable angina (UA) is still unknown. Thirty-seven consecutive patients with recent chest pain or discomfort were recruited. Patients with both Braunwald's class IB-IIIB and with coronary stenosis (or stenoses) of >75% were assigned to the UA group (n = 19, aged 67.2 +/- 8.2 years), and the rest to the control group (n = 18, aged 63.4 +/- 8.7 years). The serum levels of sCD40L and sCD40, and the plasma levels of matrix metalloproteinase (MMP)-9, were measured by enzyme-linked immunosorbent assays. A significantly inverse correlation between sCD40L and sCD40 was shown in the controls (r = -0.72, P = 0.0007), but was absent in the UA group (r = -0.16, P not significant), although there was no statistical significance between these groups in terms of serum levels of sCD40L or sCD40. The difference of the regression slopes of these regression lines was statistically significant (P < 0.01). Additionally, there was a significant correlation between sCD40 and plasma levels of MMP-9 in the patients with and without UA (r = 0.58, P = 0.0096), but no significant correlation between sCD40L and MMP-9 levels (r = 0.00, P not significant). The balance between CD40 and CD40L may be lost in patients with UA. Soluble CD40 expression may also be related to MMP-9 expression in atherosclerotic tissues. PMID- 16314906 TI - Readmission and mortality in patients undergoing off-pump coronary artery bypass surgery with fast-track recovery protocol. AB - The development of less invasive methods for myocardial revascularization such as "off-pump" cardiac surgery, and new methods of anesthesia and postoperative care protocols such as "fast-track recovery" (FTRC), have contributed to a significant reduction in postoperative intensive care unit (ICU) and hospital length of stay after cardiac surgical procedures. The objectives of this study were to identify perioperative risk factors of prolonged hospital stay, hospital mortality, and readmission rates in off-pump coronary artery bypass surgery (CABG) patients undergoing the FTRC protocol. Eighty consecutive patients undergoing off-pump coronary artery bypass surgery with FTRC protocol were included in the study. For the first purpose of this protocol, early extubation is defined as removal of the endotracheal tube within 6 h of arrival at the surgical ICU. The second purpose was to obtain a minimal length of stay in the ICU (<24 h) and hospital discharge within 5 days. We analyzed the influence of the preoperative, intraoperative, and postoperative variables on prolonged hospital stay, hospital mortality, and hospital readmission. Three patients died during hospitalization, giving a hospital mortality rate of 3.75%. The causes of hospital death were massive stroke and sepsis. Using multivariate logistic regression analysis, hypertension (P = 0.0185), postoperative stroke (P = 0.0001), and sternal infection (P = 0.0007) were identified as independent predictors of hospital mortality. Mean hospital length of stay was 4.23 +/- 0.75 days. Univariate and multivariate logistic regression analysis revealed that postoperative blood use (P = 0.0095) was the major independent predictor of prolonged hospital stay. During the 30-day observation period, seven patients were readmitted. One of these patients died on postoperative day 45 from mediastinitis and sepsis. Multivariate logistic regression analysis identified age (P = 0.0033) and hypertension (P = 0.045) as independent predictors of hospital readmission. FTRC protocols can be performed safely in patients with off-pump CABG, and the mortality and readmission rates following this protocol were found to be within acceptable ranges. PMID- 16314907 TI - Systolic time intervals revisited: correlations with N-terminal pro-brain natriuretic peptide in a community population. AB - The usefulness of automated measurements of the systolic time intervals in community screening deserves investigation. The systolic time intervals, including pre-ejection period (PEP), the left ventricular ejection time (LVET), and PEP/LVET were automatically and rapidly determined from signals of electrocardiography, phonocardiography, and pulse volume recording in 1087 residents aged > or = 40 years in a community of homogeneous Chinese. Receiver operating characteristics analysis was performed to determine the cutoff values for PEP, ET, and PEP/LVET in predicting high N-terminal pro-brain natriuretic peptide (NT-proBNP). The prevalence of high NT-proBNP was 12.3% in men and 8.3% in women. NT-proBNP was linearly related to both PEP and PEP/LVET, while there was a U-shaped relationship between NT-proBNP and LVET. In men, the sensitivity and specificity in predicting high NT-proBNP levels were 60% and 60%, respectively, for PEP > or = 89 ms; 60% and 62% for PEP/LVET > or = 0.31; and 63% and 60% for LVET < 272 ms or LVET > or = 310 ms. Various combinations of the criteria further improved either sensitivity or specificity. Women had slightly different cutoff values and performance for the various criteria of abnormal systolic time intervals and their combinations. Automated systolic time intervals appear to be useful in the screening of subjects with high NT-proBNP in a community. PMID- 16314908 TI - Evaluation of the cavotricuspid isthmus and right atrium by multidetector-row computed tomography in patients with common atrial flutter. AB - The sizes of the right atrium (RA), cavotricuspid isthmus, and Eustachian valve are predictors of success of radiofrequency catheter ablation for atrial flutter (AFL). We examined the relationship between the sizes of cavotricuspid isthmus as measured by multidetector-row computed tomography (MDCT) and fluoroscopy. We used eight-detector MDCT to measure the tricuspid isthmus of 23 patients prior to linear ablation for common AFL. One patient with a deep pouch in the RA was excluded. Parameters measured were (1) the length of the trace of isthmus (Ti), which was equivalent to the blocking line; (2) the size of the tricuspid isthmus (DTi); and (3) the distance from the tricuspid valve and inferior vena cava (IVC) (LDTi). DTi and LDTi indicate the size of the RA, reflecting the appropriately sized steerable ablation catheter, respectively. Of the 22 patients, 21 were ablated successfully without recurrence of AFL, and clinical success was achieved in one additional patient despite failure to obtain a bidirectional block. Ti, DTi, and LDTi were correlated with fluoroscopy time (r = 0.84, r = 0.88, and r = 0.88, respectively; P < 0.0001), total delivered energy (r = 0.81, r = 0.80, and r = 0.83, respectively; P < 0.0001), and application time (r = 0.84, r = 0.80, and r = 0.87, respectively; P < 0.0001). Measurement of the tricuspid isthmus by MDCT may noninvasively provide important information for successful linear ablation. PMID- 16314909 TI - Coronary pressure measurement to determine treatment strategy for equivocal left main coronary artery lesions. AB - It is often hard to select a treatment strategy for equivocal left main coronary artery (LMCA) disease. We investigated the usefulness of coronary pressure (CP) measurement for determining the treatment strategy in intermediate LMCA disease. We measured CP in 15 consecutive patients with equivocal LMCA disease (age 67.6 +/- 7.5 years, 14 males). Myocardial fractional flow reserve (FFRmyo) was obtained as the ratio of CP distal to the lesion/aortic pressure under maximal coronary dilation. Patients with FFRmyo > or = 0.75 and <0.75 received medical therapy and coronary artery bypass grafting (CABG), respectively, and were followed up for 32.5 +/- 9.7 (20-47) months. Eight patients received medical therapy and 7 patients underwent CABG in accordance with the FFRmyo criteria noted above. FFRmyo of the LMCA was 0.91 +/- 0.01 and 0.61 +/- 0.03 in patients who received medical and surgical therapy, respectively. Neither reference vessel diameter, minimal lumen diameter, nor percent diameter stenosis was significantly different between patients who received medical and surgical therapy. During the follow-up period, no patients with medical therapy showed symptoms due to the LMCA lesion. Similarly, 5 of 7 patients with CABG showed improvement of symptoms and the remaining 2 patients were hospitalized with congestive heart failure. No cardiac death was recorded in the patients with medical or surgical therapy. In conclusion, the present results clearly demonstrated that CP is clinically useful for determining the treatment strategy for equivocal LMCA lesions but coronary angiography is not. PMID- 16314910 TI - Taurine prevents myocardial ischemia/reperfusion-induced oxidative stress and apoptosis in prolonged hypothermic rat heart preservation. AB - Taurine (2-aminoethanesulfonic acid) is a potent antioxidant and inhibits cell apoptosis in ischemic reperfusion injury. In this study we evaluated whether addition of taurine to St. Thomas' cardioplegic solution enhances its myocardial protective effects in prolonged hypothermic heart preservation in rats. Hearts isolated from male Sprague-Dawley rats were mounted on a Langendorff apparatus to estimate baseline cardiac function, then arrested and stored in St. Thomas' cardioplegic solution, with taurine (10 mM; taurine group, n = 8) or without taurine (control group, n = 8), for 6 h at 4 degrees C. After storage, the hearts were reperfused and heart rate (HR), coronary flow (CF), left ventricular developed pressure (LVP), and positive maximum left ventricular developing pressure (max LV dp/dt) were measured. The LV tissue was examined immunohistochemically for determining DNA oxidative stress and cell apoptosis. Compared with control groups, recovery of LVP (P < 0.001), max LV dp/dt (P < 0.001), and coronary flow (P < 0.001) were significantly enhanced, whereas glutamic oxaloacetic transaminase (P < 0.01), lactate dehydrogenase (P < 0.05), creatine phosphate kinase (P < 0.01), 8-hydroxy-2'-deoxyguanosine index (P < 0.01), caspase-3 mRNA expression (P < 0.05), and percentage of TUNEL-positive cardiomyocytes (P < 0.05) were reduced in the taurine group. Addition of taurine to St. Thomas' cardioplegic solution improved cardiac function recovery for prolonged hypothermic rat heart preservation by suppressing DNA oxidative stress and cell apoptosis. PMID- 16314911 TI - Clinical characteristics and diagnosis of double-orifice left atrioventricular valve associated with other congenital heart disease. AB - Four cases of double-orifice left atrioventricular (AV) valve are reported. Three of the four patients with double-orifice left AV valve had other associated congenital heart diseases, and the fourth had double-orifice left AV valve alone. A patient with associated ventricular septal defect, who presented with pulmonary congestion and hypertension, suffered from severe heart failure. However, these symptoms improved with the spontaneous closure of the ventricular septal defect. The patient with double-orifice left AV valve alone showed no cardiac symptoms. Two-dimensional and Doppler echocardiography did not detect the double-orifice left AV valve in two of the four patients. More recently, the echocardiographic technique has allowed a noninvasive and more frequent detection of this abnormality. But both of these patients had left-to-right atrial shunt and subsequent reduced transmitral flow and left ventricular volume, which may have made it difficult to detect the morphological and hemodynamic characteristics of double-orifice left AV valve. Careful and repeated echocardiographic observation of mitral configuration is required to determine the presence of double-orifice left AV valve when heart disease is associated with a left-to-right atrial shunt. PMID- 16314912 TI - Plasma homocysteine and lipoprotein (a) levels in Turkish patients with metabolic syndrome. AB - High serum total homocysteine (tHcy) and lipoprotein (a) [Lp(a)] levels are independent risk factors for cardiovascular disease. In this study, we examined the relationship of tHcy and Lp(a) levels with the components of metabolic syndrome. Fifty-one patients diagnosed with metabolic syndrome (median age: 38 [range 25-48] years) and 50 healthy subjects (median age: 35 [26-48] years) were included in the study. We used the National Cholesterol Education Program criteria to define metabolic syndrome. Total tHcy concentrations were measured by using an IMX (Abbott Diagnostics, Abbott Park, IL, USA). Lipoprotein (a) was measured by immunonephelometry using Behring nephrometer method (Behring BN 100, Behring, Germany). Total homocysteine and Lp(a) levels were found to be higher in the metabolic syndrome group than in the control group (tHcy: 24.2 vs 13.4 micromol/l, P < 0.01 and Lp(a): 34.9 vs 15.8 mg/dl, P < 0.01). Vitamin B12 levels were lower in the metabolic syndrome group than in the control group (214 pg/ml vs 247 pg/ml, P < 0.01). In partial correlation, tHcy and Lp(a) concentrations were unrelated to metabolic syndrome or to the components of metabolic syndrome, including fasting serum triglycerides, HDL-cholesterol, fasting glucose, blood pressure, or body mass index. tHcy levels were strongly related only to the vitamin B12 concentration. The risk of cardiovascular disease is higher in patients with metabolic syndrome compared with the normal population. High tHcy and Lp(a) levels should be evaluated in this group of patients in addition to the evaluation of the parameters of metabolic syndrome. PMID- 16314913 TI - Cardiac hemangioma complicated with cerebral and coronary embolization. AB - Cardiac hemangiomas are rare, benign vascular tumors of the heart. Because of their clinical manifestations, diagnosis is difficult and few surgeons can draw from extensive experience. The purpose of this study was to report an additional case of cardiac hemangioma and to analyze the unexpected aspect of this disease. In our case, echocardiography demonstrated a mass in the left ventricle. Surgical resection was done using cardiopulmonary bypass. Histopathological examination revealed that the tumor was a hemangioma. The short-term outcome was favorable. PMID- 16314914 TI - Diagnosis of anomalous origin of the right coronary artery using multislice computed tomography: evaluation of possible causes of myocardial ischemia. AB - Anomalous origin of the right coronary artery (RCA) is a rare condition, but may cause myocardial ischemia and sudden death. Multislice computed tomography, which allows three-dimensional visualization of the coronary artery with high spatial resolution, may be the most promising imaging modality for diagnosing this anomaly. We describe a patient with anomalous origin of the RCA arising from the left sinus of Valsalva. Volume rendering, and axial and curved multiplanar images showed stenosis in the proximal portion of the RCA that coursed between the aorta and the pulmonary artery, and an acute angled take-off of the RCA from the aorta. Three-dimensional virtual angioscopic images showed a hypoplastic RCA orifice and luminal narrowing in the proximal portion of the RCA. Multislice computed tomography was thought to be useful for detecting anomalous origin of the RCA and for evaluating possible causes of myocardial ischemia. PMID- 16314915 TI - Kyphosis reduction and the rate of cement leaks after vertebroplasty of intravertebral clefts. AB - To assess the results of vertebroplasty in patients with intravertebral clefts compared to patients with normal osteoporotic fractures, we evaluated the pre- and postoperative images and pain scores (VAS) of 114 patients with 192 vertebroplasty procedures treated between March 2002 and February 2005. Intravertebral clefts were identified on conventional radiographs, MR or CT images as gas- or fluid-filled spaces adjacent to an endplate of a fractured vertebra. Forty-four vertebrae showed intravertebral clefts. All clefts were filled with PMMA showing a typical filling pattern. Due to the prone positioning of the patient during vertebroplasty, a significant reduction of the kyphosis angle was achieved in the cleft group. Cement leakage occurred in 18.2% of clefts and 46% of regular osteoporotic fractures. In all patients, good filling of the cleft was achieved no matter where the needle tip was placed in the vertebra. The VAS score was 9.1 preoperatively, 3.6 before discharge and 3.9 6 months postoperatively, showing no significant difference between both groups. Patients with intravertebral clefts show a significant reduction of the kyphosis angle compared to non-cleft patients and have a significantly lower risk of experiencing cement leakage during vertebroplasty. Pain reduction is the same in both groups. PMID- 16314916 TI - Investigation of a Kubo-formula-based approach to estimate DNA conductance in an atomistic model. AB - A novel approach to estimate DNA conductance based upon Kubo formula is presented and discussed. Using this approach, the effects of base pair mismatches, different conformational changes and base pair sequence on DNA electrical properties were investigated. The results were compared with the data from other methods. The new approach makes possible very fast estimation of conductance spectra for oligonucleotides with hundreds of base pairs and can easily be extended to treat arbitrary chemical modifications of DNA. PMID- 16314917 TI - A structure-activity study for the inhibition of metalloproteinase-9 activity and gene expression by analogues of gallocatechin-3-gallate. AB - Catechins are able to modulate the gelatinolytic activity of matrix metalloproteinase-9 (MMP-9) by reducing its release from macrophages. Gallocatechins decrease MMP-9 secretion by lowering MMP-9 promoter activity and mRNA levels. The effect appears to be dependent on some structural and stereochemical requirements. In this study, the relationship between chemical structure and activity was studied by testing the effect of analogues of (+/-) gallocatechin-3-gallate (+/-)-GCG, selectively deprived of hydroxyl groups, on MMP-9 activity, transcription, and secretion. Our results indicate that (+/-)-GCG and (+/-)-catechin-3-gallate are characterized by a substitution pattern compatible with direct inhibition of MMP-9 activity. Conversely, when transcription was the target, (+/-)-trans-3-flavanol-3-benzoate, lacking all the hydroxyl groups, was the most effective both in lowering MMP-9 promoter activity and consequently protein secretion, and in inhibiting nuclear-factor-kappaB driven transcription. Our results suggest that the structural requirements for enzyme inhibition are different from those necessary for targeting gene expression. PMID- 16314918 TI - Chaperone-like features of bovine serum albumin: a comparison with alpha crystallin. AB - The chaperone behaviour of bovine serum albumin was compared with that of alpha crystallin. The chaperone activity was assessed by measuring: (i) the ability to antagonize protein aggregation induced by heat; (ii) the capability to protect the activity of thermally stressed enzymes and (iii) the effectiveness in assisting the functional recovery of chemically denatured sorbitol dehydrogenase. Despite the lack of structural analogies, both proteins show several functional similarities in preventing inactivation of thermally stressed enzymes and in reactivating chemically denatured sorbitol dehydrogenase. As with alpha crystallin, the chaperone action of bovine serum albumin appears to be ATP independent. Bovine serum albumin appears significantly less effective than alpha crystallin only in preventing thermally induced protein aggregation. A possible relationship between chaperone function and structural organization is proposed. Together, our results indicate that bovine serum albumin acts as a molecular chaperone and that, for its particular distribution, can be included in the extracellular chaperone family. PMID- 16314919 TI - Glucocorticoids in T cell apoptosis and function. AB - Glucocorticoids (GCs) are a class of steroid hormones which regulate a variety of essential biological functions. The profound anti-inflammatory and immunosuppressive activity of synthetic GCs, combined with their power to induce lymphocyte apoptosis place them among the most commonly prescribed drugs worldwide. Endogenous GCs also exert a wide range of immunomodulatory activities, including the control of T cell homeostasis. Most, if not all of these effects are mediated through the glucocorticoid receptor, a member of the nuclear receptor superfamily. However, the signaling pathways and their cell type specificity remain poorly defined. In this review, we summarize our present knowledge on GC action, the mechanisms employed to induce apoptosis and the currently discussed models of how they may participate in thymocyte development. Although our knowledge in this field has substantially increased during recent years, we are still far from a comprehensive picture of the role that GCs play in T lymphocytes. PMID- 16314920 TI - Increased mitochondrial palmitoylcarnitine/carnitine countertransport by flavone causes oxidative stress and apoptosis in colon cancer cells. AB - Cancer cell metabolism is characterized by limited oxidative phosphorylation in order to minimize oxidative stress. We have previously shown that the flavonoid flavone in HT-29 colon cancer cells increases the uptake of pyruvate or lactate into mitochondria, which is followed by an increase in O(2) (-.). production that finally leads to apoptosis. Similarly, a supply of palmitoylcarnitine in combination with carnitine induces apoptosis in HT-29 cells by increasing the mitochondrial respiration rate. Here we show that flavone-induced apoptosis is increased more than twofold in the presence of palmitoylcarnitine due to increased mitochondrial fatty acid transport and the subsequent metabolic generation of O(2) (-.) in mitochondria is the initiating factor for the execution of apoptosis. PMID- 16314921 TI - The parvins. AB - The parvins are a family of proteins involved in linking integrins and associated proteins with intracellular pathways that regulate actin cytoskeletal dynamics and cell survival. Both alpha-parvin (PARVA) and beta-parvin (PARVB) localize to focal adhesions and function in cell adhesion, spreading, motility and survival through interactions with partners, such as integrin-linked kinase (ILK), paxillin, alpha-actinin and testicular kinase 1. A complex of PARVA with ILK and the LIM protein PINCH-1 is critical for cell survival in a variety of cells, including certain cancer cells, kidney podocytes and cardiac myocytes. While PARVA inhibits the activities of Rac1 and testicular kinase 1 and cell spreading, PARVB binds alphaPIX and alpha-actinin, and can promote cell spreading. In contrast to PARVA, PARVB inhibits ILK activity and reverses some of its oncogenic effects in cancer cells. This review focuses on the structure and function of the parvins and some possible roles in human diseases. PMID- 16314922 TI - The Membrane Protein Data Bank. AB - The Membrane Protein Data Bank (MPDB) is an online, searchable, relational database of structural and functional information on integral, anchored and peripheral membrane proteins and peptides. Data originates from the Protein Data Bank and other databases, and from the literature. Structures are based on X-ray and electron diffraction, nuclear magnetic resonance and cryoelectron microscopy. The MPDB is searchable online by protein characteristic, structure determination method, crystallization technique, detergent, temperature, pH, author, etc. Record entries are hyperlinked to the PDB and Pfam for viewing sequence, three dimensional structure and domain architecture, and for downloading coordinates. Links to PubMed are also provided. The MPDB is updated weekly in parallel with the Protein Data Bank. Statistical analysis of MPDB records can be performed and viewed online. A summary of the statistics as applied to entries in the MPDB is presented. The data suggest conditions appropriate for crystallization trials with novel membrane proteins. PMID- 16314924 TI - Understanding microtubule dynamics for improved cancer therapy. AB - Microtubules (MTs), key components of the cytoskeleton, are dynamic polymers of tubulin that form a well-organized network of polarized tube filaments. MT dynamics are highly regulated both spacially and temporally by several MT-related proteins, themselves regulated by several kinases and phosphatases via signaling cascades, and also by coordinated interactions with actin cytoskeleton and adhesion sites. Regulation of MT dynamics is crucial for mitosis, cell migration, cell signaling and trafficking. MT-targeted drugs (MTDs), which constitute a major anticancer drug family with antimitotic and antiangiogenic properties, inhibit tumor progression mainly by altering MT dynamics in both cancer and endothelial cells. Identification of proteins regulating the MT network will lead to a better understanding of tumor progression regulators and will be helpful in improving cancer therapy. PMID- 16314925 TI - Functionally independent subunits in the oligomeric structure of the GABA cotransporter rGAT1. AB - We have combined structural and functional approaches to investigate the role of oligomerization in the operation of the GABA transporter rGAT1. Xenopus laevis oocytes were induced to express, either separately or simultaneously, the wild type form of rGAT1 and a mutated (Y140W) form, unable to translocate GABA and to generate transport currents, although its intramembrane charge movement properties are only slightly affected. These characteristics, together with the insensitivity of Y140W to the blocking action of SKF89976A, were used to study the possible functional interaction of the two forms in an heteromeric structure. The electrophysiological data from oocytes coexpressing wild-type and Y140W rGAT1 were consistent with a completely independent activity of the two forms. Oligomerization was also studied by fluorescence resonance energy transfer (FRET) in tsA201 cells expressing the transporters fused with cyan and yellow fluorescent proteins (ECFP and EYFP). All combinations tested (WT-ECFP/WTEYFP, Y140W-ECFP/Y140W-EYFP and WT-ECFP/ Y140W-EYFP) were able to give rise to FRET, confirming the formation of homo- as well as heterooligomers. We conclude that, although rGAT1 undergoes structural oligomerization, each monomer operates independently. PMID- 16314923 TI - Diversity of Cl(-) channels. AB - Cl(-) channels are widely found anion pores that are regulated by a variety of signals and that play various roles. On the basis of molecular biologic findings, ligand-gated Cl(-) channels in synapses, cystic fibrosis transmembrane conductors (CFTRs) and ClC channel types have been established, followed by bestrophin and possibly by tweety, which encode Ca(2+)-activated Cl(-) channels. The ClC family has been shown to possess a variety of functions, including stabilization of membrane potential, excitation, cell-volume regulation, fluid transport, protein degradation in endosomal vesicles and possibly cell growth. The molecular structure of Cl(-) channel types varies from 1 to 12 transmembrane segments. By means of computer-based prediction, functional Cl(-) channels have been synthesized artificially, revealing that many possible ion pores are hidden in channel, transporter or unidentified hydrophobic membrane proteins. Thus, novel Cl(-)-conducting pores may be occasionally discovered, and evidence from molecular biologic studies will clarify their physiologic and pathophysiologic roles. PMID- 16314926 TI - Understanding the importance of selenium and selenoproteins in muscle function. AB - Selenium is an essential trace element. In cattle, selenium deficiency causes dysfunction of various organs, including skeletal and cardiac muscles. In humans as well, lack of selenium is associated with many disorders, but despite accumulation of clinical reports, muscle diseases are not generally considered on the list. The goal of this review is to establish the connection between clinical observations and the most recent advances obtained in selenium biology. Recent results about a possible role of selenium-containing proteins in muscle formation and repair have been collected. Selenoprotein N is the first selenoprotein linked to genetic disorders consisting of different forms of congenital muscular dystrophies. Understanding the muscle disorders associated with selenium deficiency or selenoprotein N dysfunction is an essential step in defining the causes of the disease and obtaining a better comprehension of the mechanisms involved in muscle formation and maintenance. PMID- 16314927 TI - Transgene silencing may be mediated by aberrant sense promoter sequence transcripts generated from cryptic promoters. AB - To investigate whether silencing of a T-DNA-carried nptII gene in five single copy transgenic Arabidopsis lines might be due to position effects, genomic DNA flanking the insertions was analysed for gene density, GC content, presence of short repeats and transposable elements, i.e. factors suggested to promote silencing. No single, common factor could explain the observed silencing. However, in two lines, a transcript covering the nos promoter driving the nptII gene was detected. In sibling sublines with approximately 100% silencing, the nos promoter was heavily methylated. In silico analysis suggested the presence of cryptic core promoters upstream of the nos promoter, in one case in the plant DNA and in the other in a short inverted TDNA region. These fragments were able to drive reporter gene expression in transgenic Arabidopsis plants. Our results indicate that methylation and silencing of transgenic promoters may be mediated by aberrant RNA transcribed from cryptic promoters at the transgene insertion site. PMID- 16314928 TI - Divergent effects of the major mast cell products histamine, tryptase and TNF alpha on human fibroblast behaviour. AB - Fibroblast proliferation is a key process in tissue remodeling and mast cells (MCs) are thought to play a crucial role. Having established that the three major MC products, tryptase, histamine and TNF-alpha (TNF) are normally present in human skin MCs, which are in close proximity to dermal fibroblasts, we studied their individual effects on cell cycle-controlled human dermal fibroblasts (HFFF2). These cells express receptors (H1, PAR2, TNFR1/2) for the major MC mediators, but only tryptase or a PAR2 agonist peptide stimulated proliferation and gene expression. TNF was antimitotic, and histamine, while elevating intracellular Ca2+ levels at high concentrations, did not affect proliferation. We conclude that MC products but also composition and numbers of respective receptors on fibroblasts are crucially responsible for fibroproliferative events. PMID- 16314929 TI - Conotoxins and the posttranslational modification of secreted gene products. AB - The venoms of predatory cone snails (genus Conus) have yielded a complex library of about 50-100,000 bioactive peptides, each believed to have a specific physiological target (although peptides from different species may overlap in their target specificity). Conus has evolved the equivalent of a drug development strategy that combines the accelerated evolution of toxin sequences with an unprecedented degree of posttranslational modification. Some Conus venom peptide families are the most highly posttranslationally modified classes of gene products known. We review the variety and complexity of posttranslational modifications documented in Conus peptides so far, and explore the potential of Conus venom peptides as a model system for a more general understanding of which secreted gene products may have modified amino acids. Although the database of modified conotoxins is growing rapidly, there are far more questions raised than answers provided about possible mechanisms and functions of posttranslational modifications in Conus. PMID- 16314930 TI - Microbial life at high temperature, the challenges, the strategies. PMID- 16314931 TI - Solute traffic across mammalian peroxisomal membrane--single channel conductance monitoring reveals pore-forming activities in peroxisomes. AB - Mouse liver peroxisomes were isolated by centrifugation in a self-generated Percoll gradient followed by an Optiprep density gradient centrifugation. Peroxisomes contributed 90-96% of the total protein content in the fraction, as confirmed by marker enzyme assays, protein pattern in SDS-PAGE, immunoblotting, and electron microscopy. Solubilized peroxisomal membrane proteins were reconstituted into a planar lipid bilayer. A single-channel conductance monitoring of the reconstituted lipid bilayer revealed the presence of two pore forming components with a conductance in 1 M KCl of 1.3 nS and 2.5 nS. Control experiments with fractions enriched in mitochondria, lysosomes, and fragments of endoplasmic reticulum showed that the peroxisomal channel-forming activities were not due to admixture of isolated peroxisomes with other cellular organelles. The peroxisomal channels were well preserved in membrane preparations but became unstable after solubilization from the membranes by detergent. PMID- 16314932 TI - Mucosal effector memory T cells: the other side of the coin. AB - Immunological memory allows for rapid and effective protective immunity to previously encountered pathogens. New insights in understanding specific memory differentiation and function have now indicated that in addition to providing enhanced immunity, an important purpose of immunological memory is to provide immediate protection at all sites of the body, including non-lymphoid tissues. Effector memory CD8 T cells have the capacity to reside long-term at epithelial surfaces, where they allow for rapid containment of the invading pathogens at the local entry site and prevent systemic spreading and excessive immune responses. The accumulation of tissue-specific memory T cell subsets, together with cross reactivity of these antigen-experienced T cells even to unrelated pathogens, provides flexibility and expansion of their specificity repertoire that over time greatly surpasses that of the declining naive T cell populations. This review will discuss new insights into T cell memory. We will focus in particular on the generation and function of effector memory CD8 T cells at the intestinal mucosa, which represents one of the largest entry sites for pathogens. PMID- 16314933 TI - siRNA-mediated transcriptional gene silencing: the potential mechanism and a possible role in the histone code. AB - Epigenetics is the study of meiotically and mitotically heritable changes in gene expression which are not coded for in the DNA. Three distinct mechanisms appear intricately related in initiating and sustaining epigenetic modifications: RNA associated silencing, DNA methylation and histone modification. Recently, in human cells small-interfering RNAs (siRNAs) have been shown to mediate transcriptional gene silencing (TGS). The observation that siRNAs can function to suppress gene expression at the level of transcription has created a major paradigm shift in mammalian RNA interference. The putative mechanism(s) of siRNA mediated TGS in both yeast and human cells will be discussed. Undoubtedly, the ramifications from this paradigm shift in which RNA has demonstrated a potent and specific capability to regulate the expression of the gene are immeasurable both therapeutically (i. e. directed control of gene expression) and biologically in understanding the evolution of the cell. PMID- 16314934 TI - Functionality of the TRPV subfamily of TRP ion channels: add mechano-TRP and osmo TRP to the lexicon! AB - Transient receptor potential (TRP) ion channels have been identified as cellular sensors responding to diverse external and internal stimuli. This review will cover the TRPV subfamily that comprises vertebrate and invertebrate members. The six mammalian TRPV channels were demonstrated to function in thermosensation, mechanosensation, osmosensation and Ca(2+) uptake. Invertebrate TRPV channels, five in Caenorhabditis elegans and two in Drosophila, have been shown to play a role in mechanosensation, such as hearing and proprioception in Drosophila and nose touch in C. elegans, and in the response to osmotic and chemical stimuli in C. elegans. We will focus here on the role that TRPV ion channels play in mechanosensation and a related sensory (sub-)modality, osmosensation. PMID- 16314935 TI - GTPase activating proteins: structural and functional insights 18 years after discovery. AB - The conversion of guanosine triphosphate (GTP) to guanosine diphosphate (GDP) and inorganic phosphate (P(i)) by guanine nucleotide binding proteins (GNBPs) is a fundamental process in living cells and represents an important timer in intracellular signalling and transport processes. While the rate of GNBP-mediated GTP hydrolysis is intrinsically slow, direct interaction with GTPase activating proteins (GAPs) accelerates the reaction by up to five orders of magnitude in vitro. Eighteen years after the discovery of the first GAP, biochemical and structural research has been accumulating evidence that GAPs employ a much wider spectrum of chemical mechanisms than had originally been assumed, in order to regulate the chemical players on the catalytic protein-protein interaction stage. PMID- 16314936 TI - Improvement of functional outcome after radical surgery in glioblastoma patients: the efficacy of a navigation-guided fence-post procedure and neurophysiological monitoring. AB - This retrospective study investigated the functional outcomes of patient with glioblastoma receiving radical surgery before and after the adoption of the navigation-guided fence-post (NGFP) procedure and neurophysiological monitoring. We investigated 42 glioblastoma patients receiving radical surgery in our institute between 1980 and 2005. Of the 42 patients, 18 patients from 1980 to 1996 (1st term) underwent radical surgery without navigation system guidance, NGFP, or neurophysiological monitoring; 11 patients from 1997 to 2002 (2nd term) underwent surgery with simple navigation system guidance but without NGFP procedure or neurophysiological monitoring, and 13 patients from 2003 to 2005 (3rd term) underwent surgery with the NGFP procedure and neurophysiological monitoring as appropriate. There were no significance differences between any of the three term groups in age, gender, preoperative KPS score, or 'surgical staging for glioma' according to the difficulty of surgery. The rates of 95% or greater volume reduction in each term were 38.9%, 54.5% and 76.9%. The rates of morbidity were 38.9%, 18.1% and 0%. The change in KPS scores (delta KPS) before and after the perioperative period in each term were -16.1 +/- 6.6 SEM, -9.0 +/- 5.8 SEM and +8.5 +/- 3.7 SEM, respectively. The delta KPS in the 3rd term was significantly better than those of 1st and 2nd terms (P < 0.01, Kruskal-Wallis rank test). The rate of patients who were discharged to home and who resumed daily useful life without assistance was 38.9%, 63.6% and 84.6% in each term, respectively. The mean survival times in each term were 9.9, 14.0 and 16.8 months. The introduction of the NGFP procedure and neurophysiological monitoring in glioblastoma radical surgery improved the functional outcome of patients. PMID- 16314937 TI - The effect of thermotherapy using magnetic nanoparticles on rat malignant glioma. AB - Thermotherapy using magnetic nanoparticles is a new technique for interstitial hyperthermia and thermoablation based on magnetic field-induced excitation of biocompatible superparamagnetic nanoparticles. To evaluate the potential of this technique for minimally invasive treatment, we carried out a systematic analysis of its effects on experimental glioblastoma multiforme in a rat tumor model. Tumors were induced by implantation of RG-2-cells into the brains of 120 male Fisher rats. Animals were randomly allocated to 10 groups of 12 rats each, including controls. Animals received two thermotherapy treatments following a single intratumoral injection of two different magnetic fluids (dextran- or aminosilane-coated iron-oxide nanoparticles). Treatment was carried out on days four and six after tumor induction using an alternating magnetic field applicator system operating at a frequency of 100 kHz and variable field strength of 0-18 kA/m. The effectiveness of treatment was determined by the survival time of the animals and histopathological examinations of the brain and the tumor.Thermotherapy with aminosilane-coated nanoparticles led up to 4.5-fold prolongation of survival over controls, while the dextran-coated particles did not indicate any advantage. Intratumoral deposition of the aminosilane-coated particles was found to be stable, allowing for serial thermotherapy treatments without repeated injection. Histological and immunohistochemical examinations after treatment revealed large necrotic areas close to particle deposits, a decreased proliferation rate and a reactive astrogliosis adjacent to the tumor.Thus, localized interstitial thermotherapy with magnetic nanoparticles has an antitumoral effect on malignant brain tumors. This method is suitable for clinical use and may be a novel strategy for treating malignant glioma, which cannot be treated successfully today. The optimal treatment schedules and potential combinations with other therapies need to be defined in further studies. PMID- 16314938 TI - Hypofractionated radiotherapy boost for dose escalation as a treatment option for high-grade spinal cord astrocytic tumor. AB - PURPOSE: To retrospectively analyze the outcome of post-operative radiotherapy for spinal cord glioma with the emphasis on the hypofractionated radiotherapy boost for dose escalation as a treatment option for high-grade spinal cord astrocytic tumors. MATERIALS AND METHODS: Forty-one patients with spinal cord glioma received post-operative radiotherapy between 1979 and 2003. The median age was 34 years (range, 10-66 years). Median follow-up was 49 months (range, 5-291 months). There were 12 low-grade astrocytic tumors, 11 high-grade astrocytic tumors, 16 low-grade ependymal tumors and 2 high-grade ependymal tumors. Among 11 patients with high-grade astrocytic tumors, 5 with anaplastic astrocytoma and 1 with glioblastoma received hypofractionated radiotherapy boost for dose escalation. The median total dose of the conventional radiotherapy was 45.5 Gy in 19 fractions (range, 30.0-60.0 Gy). The median normalized total dose (using daily dose of 2.0 Gy and an alpha/ per thousandbeta ratio of 2.0) of the hypofractionated radiotherapy boost was 131 Gy2 (range, 85-249). RESULTS: The Kaplan-Meier survival rates at 10 years from the date of the first surgery were 64% for the entire group, 47% for the astrocytic tumors and 84% for the ependymal tumors, respectively (P=0.009). Among 11 patients with high-grade astrocytic tumors, the actuarial survival rate at 10 years was 35%. The actuarial survival rates at 10 years were 67% for those who received hypofractionated radiotherapy boost for dose escalation, and 20% for those who did not (P=0.47). DISCUSSION: The results for ependymal tumors and low-grade astrocytic tumors were comparable to those reported in the literature. Hypofractionated radiotherapy boost for dose escalation may help to prolong the survival of patients with high-grade astrocytic tumors. PMID- 16314939 TI - Cell type- and region-dependent coxsackie adenovirus receptor expression in the central nervous system. AB - Model systems have shown that adenoviral vector mediated transient gene expression can potentially be applied for the treatment of brain tumours, neurodegenerative diseases and brain injuries. Most studies utilized adenovirus serotype 5 (Ad5) based vectors, which as adhesion molecules require the coxsackie adenovirus receptor (CAR) as a critical determinant for cellular infection. In this report, we have systematically characterized CAR expression in the adult human central nervous system (CNS) by using immunohistochemistry. A total of 85 specimens from various CNS regions were investigated for CAR expression in a cell type-dependent context. The most marked staining positivity was found in the choroid plexus and the pituitary gland. The neocortex had scattered positive neurons, while the white matter was mainly negative. We need to consider the possible adverse effects and the possible damage caused by adenoviral gene therapy if the virus-vector also binds to normal brain cells. PMID- 16314940 TI - Development of contrast enhancement after long-term observation of a dysembryoplastic neuroepithelial tumor. AB - Dysembryoplastic neuroepithelial tumors (DNET) are usually benign lesions that arise in cortical regions and are discovered after new onset of seizure. These lesions have many different imaging characteristics. We report a patient with a presumed low-grade medial temporal lobe lesion that was followed for many years without any change in size or imaging characteristics. This previously non enhancing tumor evolved to become contrast enhanced on routine imaging without apparent tumor growth. The patient underwent surgery, and the pathology was confirmed as a DNET with no atypical changes. This case demonstrates the potential that DNETs may exhibit a changing MRI pattern over time. Natural history, imaging characteristics, and management are reviewed. PMID- 16314941 TI - Response to chemotherapy of a radiation-induced glioblastoma multiforme. AB - BACKGROUND: Radiation-induced glioblastoma multiforme (GBM) is particularly resistant to treatment and therapeutic options are limited. We report a patient with a radiation-induced GBM who had a complete response to carmustine and survived for 44 months. PATIENTS AND METHODS: Case report of a 38-year-old man with a radiation-induced GBM that responded to carmustine. RESULTS: Our patient developed a left occipital GBM 35 years after a left cerebellar astrocytoma was treated with surgery and radiation therapy (4500 rad). The GBM was treated with subtotal resection followed by four cycles of carmustine; a complete response was achieved. He relapsed 34 months after diagnosis and with further surgery survived 44 months from his diagnosis of GBM. CONCLUSION: GBMs may be a late complication of radiation treatment for pediatric brain tumors. If further radiotherapy is not a therapeutic option, chemotherapy may result in prolonged survival. PMID- 16314942 TI - Second primary tumor in anti-Ma1/2-positive paraneoplastic limbic encephalitis. AB - Memory loss can be a symptom of paraneoplastic limbic encephalitis (PLE) a neuropsychiatric disorder associated mostly with small-cell lung cancer and anti Hu antibodies or with testicular tumors and anti-Ma2 antibodies. We present the case of a patient with temporal coincidence of beginning cognitive decline and diagnosis of a carcinoma of the prostate in whom we diagnosed anti-Ma1/Ma2 positive PLE. The tumor had been completely resected but memory impairment further deteriorated. As the effective treatment of the cancer is considered as the most efficient treatment of a paraneoplastic neurological syndrome (PNS) a second neoplasia was suspected in the patient. By the aid of whole body positron emission tomography with 18-fluorine fluoro-2-deoxy-glucose (FDG-PET) an adenocarcinoma of the cecum could be detected. Two months after surgery anti-Ma antibodies were negative. We conclude that a second neoplasia should be considered, if effective cancer treatment does not lead to improvement or stabilisation of a PNS. Tumor search should be exhaustive and include PET when conventional imaging fails to show a malignancy. PMID- 16314943 TI - Immunotoxin pharmacokinetics: a comparison of the anti-glioblastoma bi-specific fusion protein (DTAT13) to DTAT and DTIL13. AB - DTAT13, a novel recombinant bispecific immunotoxin (IT) consisting of truncated diphtheria toxin, an amino-terminal (AT) fragment of the urokinase-type plasminogen activator (uPa), and a fragment of human IL-13 was assembled in order to target receptors on glioblastoma multiforme (GBM) and its associated neovasculature. Previous in vitro studies confirmed the efficacy of DTAT13 against various GBM cell lines expressing both IL-13 receptor or uPA receptor, and previous in vivo testing demonstrated the efficacy of DTAT13 in significantly inhibiting a range of xenograft tumors and showed that DTAT13 was 160- and 8-fold less toxic to the parental fusion IT, DTAT and DTIL13, respectively. To further understand the properties of DTAT13, pharmacokinetic/biodistribution experiments were performed. Binding analysis revealed that the IL-13 domain functioned independently of the uPA domain and that the K (d) for each binding domain was essentially the same as that of DTIL13 and DTAT. Flow cytometry studies indicated that DTAT13 bound better than DTAT or DTIL13. Analysis of the rate of protein synthesis inhibition in U87 MG cells by DTAT13 compared to DTAT revealed a faster rate of inhibition with DTAT13 compared to DTAT. The rate of protein synthesis inhibition of DTAT13 was identical to that of DTIL13 in U373 MG cells. Intracranial biodistribution studies revealed that DTAT13 was able to cross to the contralateral hemisphere unlike DTIL13 but similar to DTAT. These studies show that DTAT13 has properties encompassing those of both DTIL13 and DTAT and warrants further consideration for clinical development. PMID- 16314944 TI - Postoperative radiotherapy in cranial ganglioglioma. AB - We performed an institutional database search for patients with ganglioglioma in order to evaluate postoperative radiotherapy in the light of our experience. This search identified 5 patients with a mean age of 23.3 (range, 9-54 years) treated between April 1994 and April 2003. The latter date was chosen to allow a minimum follow-up of 1 year at the time of our analysis. Median follow up was 7 years (range, 1.5-11 years). Gross total tumor resection could only be achieved in one case, while other four patients had been referred after subtotal tumor excisions. All patients were treated postoperatively with conventionally fractionated (1.8-2 Gy/day) external beam radiotherapy (EBRT) to a total dose of 54-60 Gy. Patients were treated with megavoltage beams using Co(60 )or 6-MV photons. None of the patients were lost to follow up with a median follow up time of 80 months (range, 5.5-122.5 months) and all patients were alive with no evidence of disease at last follow-up. We believe that long term follow-up is necessary to observe the relapse patterns and the significance of postoperative radiotherapy needs to be further evaluated with large patient numbers to remark a conclusive statement. PMID- 16314945 TI - 3-Tesla intraoperative MR imaging for neurosurgery. AB - Intraoperative magnetic resonance (MR) image-guided neurosurgery has been performed since 1994. Using a 1.5-Tesla (T) intraoperative MR imaging system, we have performed more than 750 interventional procedures. Having validated the safety and efficacy of this surgical technique that is relatively amenable to nearly all new in-hospital MR suites, we sought to adapt this approach at our sister hospital where a new short-bore 3-T MR suite was being installed. Using many of the lessons learned from our initial experience at 1.5-T, we designed a new interventional suite that would enable surgery to be performed entirely within a 3-T MR environment. All surgical instrumentation including electrocautery, fiberoptic headlamp, power drill, and ultrasonic aspirator was entirely MR-compatible. A few items with limited ferromagnetism were utilized within the magnetic field under strict precaution. From 2/04 to 7/05, those cases initially performed within the 3-T surgical suite included one drainage and reservoir placement for a cystic craniopharyngioma, five brain biopsies and two craniotomies; one for open brain biopsy and another for lesion resection. The craniopharyngioma was successfully aspirated and had the reservoir catheter placed within the cyst. All five brain biopsies yielded diagnostic tissue. The craniotomy for mass resection demonstrated radiation necrosis. Although the metallic artifact from the biopsy needle was more prominent than at 1.5-T, accurate image interpretation was possible. Surgical needles, disposable scalpel, disposable razor, and surgical stapler were minimally ferromagnetic and safely controlled by the surgeon. There were no adverse events associated with any procedure. MR-guided neurosurgery can be safely and effectively performed at 3-T. The surgical environment at 3-T is comparable to that present at 1.5-T. PMID- 16314946 TI - Tumor growth, invasion, and angiogenesis in malignant gliomas. PMID- 16314947 TI - Postoperative cerebral abscess formation caused by Toxocara canis in a meningioma cavity. PMID- 16314948 TI - Brain metastases from apocrine carcinoma of the scalp: case report. AB - Apocrine carcinoma is an extremely rare malignant neoplasm that occurs most frequently in the axilla. Although it usually shows an indolent clinical course, it often metastasizes to regional lymph nodes and sometimes to lungs or bones. However, a literature search did not reveal any report describing the detailed clinical course of brain metastases from apocrine carcinoma. We report a case of a 54-year-old male who suffered from multiple brain metastases from apocrine carcinoma that had originated in the scalp 6 years before. The brain metastases appeared in spite of several regimens of chemotherapy for lung metastases for two years. The tumor in the right frontal lobe was successfully operated. However, the small tumor in the right occipital lobe was not cured by gamma knife surgery, and eventually required second operation. The operation had contributed to his neurologically independent life for about one year until he died for gradual progression of lung metastases. To our knowledge this is the first reported case of metastatic brain tumor from apocrine carcinoma. PMID- 16314949 TI - Osmotic blood-brain barrier disruption chemotherapy for diffuse pontine gliomas. AB - The prognosis for patients with diffuse pontine gliomas (DPG) remains poor. New aggressive innovative treatments are necessary to treat this disease. From 1984 to 1998, eight patients (4M/4F), median age 11 years, with DPG were treated with monthly osmotic blood-brain barrier disruption (BBBD) chemotherapy using intraarterial carboplatin or methotrexate and intravenous cytoxan and etoposide. Patients presented for a median duration of 6 weeks with increased intracranial pressure, long tract signs, diplopia, ataxia, and nausea/vomiting. DPG was demonstrated on magnetic resonance (MR) imaging in seven patients and on CT in one. Two patients had biopsies that showed an astrocytoma and an anaplastic astrocytoma. Three tumors enhanced on MR imaging after contrast administration. Three patients had radiation therapy before BBBD chemotherapy and four afterwards. Two patients had chemotherapy (tamoxifen, topotecan) before BBBD chemotherapy and two afterwards. In general, patients were evaluated with MR imaging every 3 months to monitor for a response to treatment. The median number of chemotherapy cycles that were administered by BBBD was 10, mean 10. Three patients also received one, two, or three cycles of intraarterial chemotherapy without BBBD. One patient that was started on carboplatin was converted to methotrexate, and five that were started on the methotrexate protocol were later converted over to carboplatin. One patient received monthly methotrexate followed by 14 days of procarbazine and one patient started on methotrexate was switched to navelbine. MR imaging demonstrated two partial responses, five patients with stable disease, and one with disease progression. The median time to tumor progression was 15 months with the range from <1 to 40 months. The median survival from the time of diagnosis was 27 months, ranging from 7 to 80 months. The median survival time from the first BBBD or intraarterial treatment was 16.5 months, ranging from 5 to 69 months. One patient was lost to follow-up with an unknown date of death. Although the sample size is small, the TTP and survival times are longer than those previously reported in other DPG series. In addition, the ability to demonstrate stable disease or partial responses in DPG on MR imaging argues for the therapeutic benefit of BBBD chemotherapy. The enhanced delivery of chemotherapy afforded by osmotic BBBD supports the further examination of this treatment modality for patients with DPG. PMID- 16314950 TI - Pathological laughter and behavioural change in childhood pontine glioma. AB - Children with pontine glioma usually present classically with ataxia, motor deficits and cranial nerve palsies. The pons has generally not been regarded as a structure that mediates complex affective behaviour. However, we report nine children who either at the time of presentation or progression demonstrated marked behavioural changes manifesting as either "pathological laughter" or separation anxiety in the form of school refusal. A mechanism of how pontine lesions can cause such complex affective and cognitive behaviour has been suggested to consist of the disruption of a network of cerebro-ponto-cerebellar pathways and the evidence for this mechanism is discussed. PMID- 16314951 TI - Human leukocyte antigen distribution analysis in North Italian brain Glioma patients: an association with HLA-DRB1*14. AB - Human leukocyte antigens (HLA) are widely expressed cell surface molecules that present antigenic peptides to T-lymphocytes and modulate the immune response against inflammatory and malignant disease. Frequently, tumoral cells express antigens that are recognized by the immune system. Ineffective immune response could be the result of defects in antigen presentation in those subjects with peculiar HLA alleles, which, owing to mechanisms that are still unknown, are unable to carry out their function. Only a few studies on glioma and HLA association have been performed to date. The aim of our study was to characterize a group of Italian Caucasian patients with glioma, to investigate a possible association between HLA antigens and cerebral glioma tumorigenesis in Italian patients. HLA typing of class I and class II loci was done by molecular typing performed on blood DNA from 36 glioma patients from northern Italy. The data obtained were compared with HLA frequencies taken from the database of northern Italian organ donors.A positive association between HLA-DRB1*14 and the presence of symptomatic cerebral glioma was observed (p = 0.02, odds ratio = 2.48, 95% confidence interval: 1.09-5.45). This is the first Italian report on a case control data study of HLA distribution conducted on a group of glioma patients and a first step in defining a possible involvement of HLA in susceptibility to brain glioma in the Italian population. PMID- 16314952 TI - Safety and efficacy of a novel cannabinoid chemotherapeutic, KM-233, for the treatment of high-grade glioma. AB - OBJECTIVE: To test in vitro and in vivo the safety and efficacy of a novel chemotherapeutic agent, KM-233, for the treatment of glioma. METHODS: In vitro cell cytotoxicity assays were used to measure and compare the cytotoxic effects of KM-233, Delta(8)-tetrahydrocannabinol (THC), and bis-chloroethyl-nitrosurea (BCNU) against human U87 glioma cells. An organotypic brain slice culture model was used for safety and toxicity studies. A human glioma-SCID mouse side-pocket tumor model was used to test in vivo the safety and efficacy of KM-233 with intratumoral and intra-peritoneal administration. RESULTS: KM-233 is a classical cannabinoid with good blood brain barrier penetration that possesses a selective affinity for the CB2 receptors relative to THC. KM-233 was as efficacious in its cytotoxicity against human U87 glioma as Delta(8)-tetrahydrocannabinol, and superior to the commonly used anti-glioma chemotherapeutic agent, BCNU. The cytotoxic effects of KM-233 against human glioma cells in vitro occur as early as two hours after administration, and dosing of KM-233 can be cycled without compromising cytotoxic efficacy and while improving safety. Cyclical dosing of KM 233 to treat U87 glioma in a SCID mouse xenograft side pocket model was effective at reducing the tumor burden with both systemic and intratumoral administration. CONCLUSION: These studies provide both in vitro and in vivo evidence that KM-233 shows promising efficacy against human glioma cell lines in both in vitro and in vivo studies, minimal toxicity to healthy cultured brain tissue, and should be considered for definitive preclinical development in animal models of glioma. PMID- 16314953 TI - Concurrent chemoradiotherapy and adjuvant chemotherapy with Topotecan for patients with glioblastoma multiforme. AB - PURPOSE: The prognosis for patients with glioblastoma multiforme remains poor. Phase II studies, meta-analyses and a phase III study show that concurrent chemoradiotherapy has an advantage over irradiation alone. In this study the effectiveness of concurrent chemoradiotherapy with Topotecan and an adjuvant chemotherapy with Topotecan was investigated. PATIENTS AND METHOD: Forty-two patients with predominantly unfavourable prognosis factors were included in the study and treated as follows: hyperfractionated accelerated radiotherapy (2x1.75Gy to 45.5 + 12.25 Gy (RP)) with a concurrent, continuous infusion of Topotecan (0.5 mg/m(2)/d, days 1-21). On day 28 the adjuvant chemotherapy (three courses) was begun according to the same scheme. RESULT: Haematological toxicities were 13/42 (30%) grade III leucopenia, 2/42 (4%) grade IV, as well as 5/42 (10%) grade III thrombopenias and 1/42 (2%) grade IV. 30/42 (71%) patients showed improvement or stabilisation of an existing neurological symptomatic complex. The median time to progression was 7.2 (+/- 0.8) months, the median total survival was 10 (+/- 1.2) months, the 2 year survival rate 4.7 (+/- 0.3)%. Prognostic factors were age, surgical radicality, performance status and the tumour volume before therapy. SUMMARY: Concurrent chemoradiotherapy and an adjuvant chemotherapy with Topotecan is feasible at acceptable toxicity levels also for patients with a moderate performance status. The patients benefit from the improvement of the clinical symptomatic complex and, even with unfavourable prognosis factors, have a higher median survival in comparison to data published on similar groups of patients given only radiotherapy. PMID- 16314954 TI - Antisense oligonucleodes targeting the focal adhesion kinase inhibit proliferation, induce apoptosis and cooperate with cytotoxic drugs in human glioma cells. AB - To examine the role of focal adhesion kinase in human glioma cells, we studied its effects on proliferation and apoptosis using FAK antisense oligonucleotide. U251 MG cells were transfected with ODNs, sense FAK, mismatch FAK and antisense FAK, respectively. Expression of FAK proteins were detected by Western blots and Immnofluoressence. Cell apoptosis and mitochondrial membrane potential were analyzed by flow cytometry. Caspase-3 activity was measured by spectrofluorometer. MTT assay was used to examine changes in cell proliferation. The protein expression of FAK in U251 MG cells decreased in antisense-FAK ODNs group significantly. Caspase-3 activity increased in cells treated with antisense FAK and down-regulated when treated with caspase-3 inhibitor. The level of cell apoptosis and loss of mitochondrial membrane potential in antisense-FAK group was higher than in the mismatch sense group. Cells proliferation was inhibited by antisense-FAK, and the effects were clearly additive when antisense oligonuceotides were added to cells treated with the anticancer agents. The results suggest that antisense-FAK ODNs inhibit U251 MG cells proliferation and induce their apoptosis. It is possible that FAK via mitochondrial and caspase-3 inhibits U251 MG cells apoptosis. And antisense oligonucleotide treatment enhances U251 MG cells sensitivity to chemotherapy. PMID- 16314955 TI - A phase II window trial of procarbazine and topotecan in children with high-grade glioma: a report from the children's oncology group. AB - The role of chemotherapy in the treatment of high-grade gliomas in children is unclear. Early reports were suggestive of improved outcome in children with high grade glioma with the addition of chemotherapy after surgery and radiation therapy. Subsequent studies did not show similar favorable contribution of chemotherapy to the outcome of these children. Further efforts to identify active chemotherapy agents in children include use of agents that have shown efficacy in adult patients with high-grade glioma and agents that have shown promise in mice bearing human xenografts of brain tumors. A Pediatric Oncology Group (POG 9431) trial tested the activity of two such agents, procarbazine and topotecan in newly diagnosed patients with high-grade glioma who had measurable disease after diagnostic surgery. Neither agent showed efficacy within the confines of the statistical design of the study. This study showed that children with high-grade glioma have an innate resistance to alkylating agents based on mismatch repair deficiency and high levels of alkyguanine transferase (AGT). Future trials should consider strategies to overcome the resistance mechanisms in children with high grade glioma. PMID- 16314956 TI - 123I-metaiodobenzylguanidine single-photon emission computerized tomography in brain tumors - a preliminary study. AB - OBJECTIVE: (123)I-metaiodobenzylguanidine (MIBG) has been developed as a functional analog of the neurotransmitter norepinephrine. The success of MIBG as an imaging agent for neural crest tumors is derived from its chemical similarities to norepinephrine. The present study aimed to explore a potential of (123)I-MIBG to differentiate embryonal tumors from other types of brain tumors. METHODS: Sixteen patients with brain tumors including three medulloblastomas, one neuroblastoma, six gliomas, and six meningiomas were examined with single-photon emission computerized tomography (SPECT) using (123)I-MIBG. The (123)I-MIBG uptake of tumors was defined as the ratios of tumor/nontumor (early and delayed T/NT) on SPECT images scanned 30 min and 6 h after intravenous injection of the tracer, respectively. Retention index was calculated as (delayed T/NT - early T/NT)/early T/NT. RESULTS: The T/NT ratios on the early images for embryonal tumors (medulloblastomas and neuroblastoma), gliomas, and meningiomas were 3.2+/ 1.7 (mean+/-SD), 1.4+/-0.3, and 1.6+/-0.5, respectively. The early uptake was significantly higher in the embryonal tumors than in gliomas (P<0.05). Delayed T/NT ratios for embryonal tumors were increased compared to the early T/NT ratios, while in contrast delayed T/NT ratios for the other tumors remained low (1.2-1.7). The high retention indices of the embryonal tumors indicate specific uptake of (123)I-MIBG in the tumors. CONCLUSION: Early high accumulation and high retention on delayed imaging may indicate a possibility of (123)I-MIBG SPECT in differentiating embryonal brain tumors from gliomas and meningiomas. PMID- 16314957 TI - Sestamibi technetium-99m brain single-photon emission computed tomography to identify recurrent glioma in adults: 201 studies. AB - OBJECT: In the follow-up of treated gliomas, CT and MRI can often not differentiate radionecrosis from recurrent tumor. The aim of this study was to assess the interest of functional imaging with (99m)Tc-MIBI SPECT in a large series of 201 examinations. METHOD: MIBI SPECT were performed in 81 patients treated for brain gliomas. A MIBI uptake index was computed as the ratio of counts in the lesion to counts in the controlateral region. SPECT was compared to stereotactic biopsy in 14 cases, or in the others cases to imaging evolution or clinical course at 6 months after the last tomoscintigraphy Two hundred and one tomoscintigraphies were performed. One hundred and two scans were true positive, 82 scans were true negative. Six scans were false positive (corresponding to 3 patients): 2 patients with an inflammatory reaction after radiosurgery, 1 with no explanation up to now. Eleven scans were false negative (5 patients): 1 patient with a deep peri-ventricular lesion, 2 patients with no contrast enhancement on MRI, 2 patients with a temporal tumor. The sensitivity for tumor recurrence was 90%, specificity 91.5% and accuracy 90.5%. We studied separately low and high grade glioma: sensitivity for tumor recurrence was respectively 91% and 89%, specificity 100% and 83% and accuracy 95% and 87%. MIBI SPECT allowed the diagnose of anaplasic degenerence of low grade sometimes earlier than clinical (5 cases) or MRI signs (7 cases). CONCLUSIONS: Our results confirm the usefullness of MIBI SPECT in the follow-up of treated gliomas for the differential diagnosis between radiation necrosis and tumor recurrence. PMID- 16314958 TI - Primary endodermal sinus tumor of the cerebellar hemisphere: a case report with review of the literature. AB - Primary intracranial endodermal sinus tumors, which have been regarded as a rare histologic subtype, usually arise in the pineal and suprasellar regions and are often associated with components of other germ cell tumors. We report an extremely rare case of pure primary endodermal sinus tumor found in the cerebellar hemisphere. A 3-year-old boy was admitted to our institution because of gait disturbance, vomiting and deteriorated mental state. MR imaging revealed the presence of a round mass with heterogeneous enhancement in the left cerebellar hemisphere. Radical surgical removal of the tumor was performed, followed by adjuvant chemotherapy, consisting of etoposide, carboplatin and bleomycin. The patient has since attended regular follow-ups, without any neurological deficit or signs of recurrence in the 4 years since diagnosis. PMID- 16314959 TI - Tetrazolium violet induces G0/G1 arrest and apoptosis in brain tumor cells. AB - Tetrazolium violet (TV), a potent anticancer agent, has been shown to induce cell growth-inhibition in tumor cells. However, the related mechanism has not been revealed yet. In this report we assessed the influence of TV on cell growth and cell cycle in brain tumor cells. Treatment of C6 tumor cells with TV (5-15 microM for 24-72 h) resulted in a growth inhibition in a dose and time-dependent manner and G0/G1 phase arrest, determined by flow cytometry analysis. These effects were accompanied by apoptosis other than necrosis, evidenced by nuclear condensation, terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay and trypan blue exclusion assay plus lactate dehydrogenase (LDH) release assay. Treatment of cells with TV at 15 microM for 24 h resulted in an increase in the activity of caspase-3, evidenced by colorimetric assay, and a dramatic up regulation of p53, accompanied with a significant increase of Bax/Bcl-2 ratio, as evidenced by immunofluorescence assay. These results suggest that TV induces growth inhibition of C6 cells through p53-midiated apoptotic pathway and G0/G1 checkpoint mechanism. Although detailed mechanisms remain to be explored, selective blockage of tumor cells in G0/G1 phase accompanied by p53-associated apoptosis makes tetrazolium violet a promising anticancer agent, meriting further investigations. PMID- 16314960 TI - Heteronymous inferior quadrantanopsia from a hypothalamic metastasis. PMID- 16314963 TI - Preparative electrophoresis with on-column optical fiber monitoring and direct elution into a minimized volume. AB - A "column-format" preparative electrophoresis device which obviates the need for gel extraction or secondary electro-elution steps is described. Separated biomolecules are continuously detected and eluted directly into a minimal volume of free solution for subsequent use. An optical fiber allows the species of interest to be detected just prior to elution from the gel column, and a small collection volume is created by addition of an ion-exchange membrane near the end of the column. PMID- 16314964 TI - High-level expression of a synthetic gene encoding a sweet protein, monellin, in Escherichia coli. AB - The expression of a synthetic gene encoding monellin, a sweet protein, in E. coli under the control of T7 promoter from phage is described. The single-chain monellin gene was designed based on the biased codons of E. coli so as to optimize its expression. Monellin was produced and accounted for 45% of total soluble proteins. It was purified to yield 43 mg protein per g dry cell wt. The purity of the recombinant protein was confirmed by SDS-PAGE. PMID- 16314965 TI - Methyl oleate modulates LIP2 expression in the lipolytic yeast Yarrowia lipolytica. AB - Methyl oleate was used as a primary carbon source and as an alternative inducer for the production of an extracellular lipase, Lip2, in Y. lipolytica strain LgX64.81 grown in a 20-l bioreactor. The lipase-encoding gene, LIP2, was investigated during culture on methyl oleate using a pLIP2-LacZ reporter fusion and we provide evidence for the involvement of methyl oleate in its regulation. PMID- 16314966 TI - Enhancement of 1,3-propanediol production by Klebsiella pneumoniae with fumarate addition. AB - Addition of 5 mM: fumarate to cultures of Klebsiella pneumoniae enhanced the rate of glycerol consumption and the production of 1,3-propanediol (PDO). Compared to the control, the activity of glycerol dehydrogenase increased by 35, 33 and 46%, the activity of glycerol dehydratase increased by 160, 210 and 115%, and the activity of 1,3-propanediol oxidoreductase increased by 25, 39 and 85% when, respectively, 5, 15 and 25 mM: fumarate were provided. At the same time, the ratio of NAD+ to NADH decreased by 20, 23 and 29%. Using a 5 l bioreactor with 5 mM: fumarate addition, the specific rate of glycerol consumption and the productivity of PDO was 30 mmol/l h and 17 mmol/l h, respectively, both increased by 35% over the control. PMID- 16314967 TI - Injectable polysaccharide microcapsules for prolonged release of minocycline for the treatment of periodontitis. AB - Injectable polysaccharide microcapsules holding minocycline were fabricated from alginate and chitosan for the treatment of periodontitis. The microcapsules were examined for the release and degradation of minocycline, as well as antimicrobial activity. The microcapsules were biodegradable and released minocycline between 10 and 1000 microg ml(-1), which was higher than the usual therapeutic concentration (1-5 microg ml(-1)), for up to 7 days. These microcapsules showed a statistically significant suppression of pathogenic bacteria, such as Prevotella intermedia causing periodontitis. The microcapsules are thus potentially useful for drug delivery for the treatment of periodontitis. PMID- 16314968 TI - Improved cellular response of osteoblast cells using recombinant human osteopontin protein produced by Escherichia coli. AB - Osteopontin is a major non-collagenous bone matrix protein secreted into the mineralizing extracellular matrix by osteoblasts during bone development. Recombinant human osteopontin (hOPN) that includes the Arg-Gly-Asp (RGD) cell recognition site was expressed in Escherichia coli and the purified osteopontin increased cell adhesion, proliferation and differentiation of osteoblast cells (p<0.05). PMID- 16314969 TI - Induction and characterization of adventitious roots directly from the explants of Panax notoginseng. AB - Adventitious roots from leafstalks and lateral roots were obtained directly from explants of Panax notoginseng. The lateral root explants were more sensitive to the induction of adventitious roots using indole-3-butyric acid. HPLC analysis of saponins extracted from the adventitious roots indicated that several protopanaxatriol saponins were present but ginsenoside Rd was missing, compared with the saponins extracted from the raw herbs. The dry weight of primary adventitious root culture of Panax notoginseng increased 5.25 times during multiplication in a classical shaking-flask system, suggesting that it is a culture system with great potential for scale-up. PMID- 16314970 TI - Rational design of thermostable lactate oxidase by analyzing quaternary structure and prevention of deamidation. AB - Our current knowledge of protein unfolding is overwhelmingly related to reversible denaturation. However, to engineer thermostable enzymes for industrial applications and medical diagnostics, it is necessary to consider irreversible denaturation processes and/or the entire quaternary structure. In this study we have used lactate oxidase (LOD), which is employed in lactic acid sensors, as a model example to design thermostable variants by rational design. Twelve mutant proteins were tested and one of them displayed a markedly greater thermostability than all the mutants we had previously obtained by random mutagenesis. This mutant was designed so as to strengthen the interaction between the subunits and stabilize the quaternary structure. Since LOD is difficult to crystallize, its three-dimensional structure remains unknown. This study shows that it is possible to carry out rational design to improve thermostability using a computer-aided quaternary structure model based on the known tertiary structure of a related protein. Critical factors required for increasing the thermal stability of proteins by rational design, where the 3-D structure is not available, are discussed. PMID- 16314971 TI - Kinetic modelling of Lactobacillus casei ssp. rhamnosus growth and lactic acid production in batch cultures under various medium conditions. AB - Enrichment of medium with yeast extract and tryptone increased growth and lactic acid production in batch cultures of Lactobacillus casei ssp. rhamnosus. A reliable kinetic model that explicitly expresses the strong relationship between microbial growth, lactic acid production and medium enrichment is provided and validated using experimental data obtained with six different medium compositions. PMID- 16314972 TI - Using redox potential to detect microbial activities during clavulanic acid biosynthesis in Streptomyces clavuligerus. AB - Production of clavulanic acid (CA) by Streptomyces clavuligerus in a shake-flask culture increased from 92 to 180 mg l(-1) with an increased O2 transfer efficiency (0.039 --> 0.058 s(-1)), which maintained the redox potential values above -250 mV. Compared with traditional measures, such as dissolved O2 concentration and respiratory activity, the redox potential can easily be determined and correlates closely with CA production. It can therefore be used to monitor microbial activities during biosyntheses of secondary metabolites. PMID- 16314973 TI - Methyl tert-butyl ether and tert-butyl alcohol degradation by Fusarium solani. AB - Fusarium solani degraded methyl tert-butyl ether (MTBE) and other oxygenated compounds from gasoline including tert-butyl alcohol (TBA). The maximum degradation rate of MTBE was 16 mg protein h and 46 mg/g protein h for TBA. The culture transformed 77% of the total carbon to 14CO2. The estimated yield for MTBE was 0.18 g dry wt/g MTBE. PMID- 16314974 TI - Biocatalytic amide reduction using Clostridium sporogenes. AB - Washed cells of Clostridium sporogenes reduced benzamide (up to 20 mM: ) to benzylamine in yields up to 73% using H2 as electron donor with less than 10 g biocatalyst/l over 24 h. Product formation exhibited complex kinetics, with a lag before benzylamine production began. Very little substrate was hydrolysed since the maximum yield of benzoic acid was only 9% of the substrate added. Boiled cells were inactivated thus confirming that amide reduction was enzyme-catalysed. PMID- 16314975 TI - Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production. AB - Adenoviruses are promising vectors for gene therapy and vaccination protocols. Consequently, the market demands for adenovirus are increasing, driving the search for new methodologies for large-scale production of concentrated vectors with warranted purity and efficacy, in a cost-effective way. Nevertheless, the production of adenovirus is currently limited by the so-called 'cell density effect', i.e. a drop in cell specific productivity concomitant with increased cell concentration at infection. Of two different serum-free culture media (CD293 and EX-Cell), evaluated for their effect on human 293 cells growth and adenovirus production at cell densities higher than 1x10(6) cells/ml, EX-Cell proved the better medium for cell growth. Although adenovirus production was equivalent in both media when the infection was performed at 1x10(6) cells/ml, at 3x10(6) cells/ml CD293 was the better. This result related to the high ammonia content in EX-Cell medium at the highest cell concentration at infection. Besides this, the large-scale production of these vectors at high cell densities often requires re feed strategies, which increase medium osmolality. While a negative effect on cell growth was observed with increasing osmolalities, adenovirus productivity was only affected for osmolalities higher than 430 mOsm. PMID- 16314976 TI - Isolation of a novel N-acetyl-D-lactosamine specific lectin from Alocasia cucullata (Schott.). AB - An N-acetyl-D: -lactosamine (LacNAc) specific lectin from tubers of Alocasia cucullata was purified by affinity chromatography on asialofetuin-linked amino activated silica. The pure lectin showed a single band in SDS-PAGE at pH 8.8 and was a homotetramer with a subunit molecular mass of 13.5 kDa and native molecular mass of 53 kDa. It was heat stable up to 55 degrees C for 15 min and showed optimum hemagglutination activity from pH 2 to 11. The lectin was affected by denaturing agents such as urea (2 M: ), thiourea (2 M: ) and guanidine-HCl (0.5 M: ) and did not require Ca2+ and Mn2+ for its activity. It was a potent mitogen at 10 microg/ml towards human peripheral blood mononuclear cells with 50% growth inhibitory potential towards SiHa (human cervix ) cancer cell line at 100 microg/ml. PMID- 16314977 TI - A new format of single chain tri-specific antibody with diminished molecular size efficiently induces ovarian tumor cell killing. AB - A combination of bi-specific antibodies (BsAb), anti-tumorxanti-CD3 and anti tumorxanti-CD28, is effective in vitro and in vivo, whereas production of two kinds of bi-specific antibodies is labor intensive and administration is complicated. Accordingly, we previously developed a new model of single chain tri specific antibody (scTsAb), sTRI, which linked both the CD3 and CD28 signals for T-cell activation in one molecule, and demonstrated its capacity for triggering T cells to kill ovary tumor cells. To improve the pharmacokinetics further and decrease the immunogenicity of scTsAb, we have now generated a new format of scTsAb, TR3H, whose molecular size is smaller than sTRI. Here we describe the construction, purification and characterization of TR3H. TR3H scTsAb bound to effector cells and tumor target cells specifically and induced redirected lyses of ovary tumor cells through freshly isolated, unstimulated human peripheral blood lymphocytes (PBLs). This new format of scTsAb possesses properties that support its potential as a new tumor immunotherapeutic agent. PMID- 16314978 TI - Production of vanillin by metabolically engineered Escherichia coli. AB - E. coli was metabolically engineered to produce vanillin by expression of the fcs and ech genes from Amycolatopsis sp. encoding feruloyl-CoA synthetase and enoyl CoA hydratase/aldolase, respectively. Vanillin production was optimized by leaky expression of the genes, under the IPTG-inducible trc promoter, in complex 2YT medium. Supplementation with glucose, fructose, galactose, arabinose or glycerol severely decreased vanillin production. The highest vanillin production of 1.1 g l(-1) was obtained with cultivation for 48 h in 2YT medium with 0.2% (w/v) ferulate, without IPTG and no supplementation of carbon sources. PMID- 16314980 TI - Embryology of the internal carotid artery dural crossing: apropos of a continuous series of 48 specimens. AB - The aim of this study was to describe the embryologic and foetal development of the anterior paraclinoid region and more precisely the relationship of the internal carotid artery to the dura mater. This has been done by examining a collection of histological sections, representing a continuous series of 48 embryologic and foetal specimens, covering the period of the first 6 months of intra-uterine life. Neurological and vascular elements develop during the embryologic period; the internal carotid artery is recognizable in the various sections of its course and acquires a histological adult parietal constitution. The foetal period corresponds to the development of the meningeal structures. The superior, medial and lateral walls appear on the fifteenth week of amenorrhoea and do not change after that. The internal carotid artery enters subarachnoid space accompanied by a sleeve of mesenchymatous cells, which fixes it to the anterior clinoid process. The constitution of this sleeve, arising from the superior wall of the lateral sellar compartment, remained independent of the principle vascular part, which allows the formation of a plan of cleavage. The foetal relations of the dura mater and the internal carotid artery were seen to be different from those of adult subjects described in the literature, suggesting an existence of period of maturation postnatally. PMID- 16314981 TI - Heterotopic ossification of the supraspinatus tendon after rotator cuff repair: case report. AB - Heterotopic ossifications of the shoulder are uncommon. Rarely, these ossifications are seen after open or even arthroscopic shoulder surgical procedures. Here, we report a patient who underwent a rotator cuff repair, complicated with an axillary nerve paralysis. Postoperatively he developed substantial ossification of the supraspinatus tendon. A review of the literature was done. To our knowledge no other cases similar to this have been reported. PMID- 16314982 TI - Racial variations in antiresorptive medication use: results from the 2000 Medical Expenditure Panel Survey (MEPS). AB - Although a number of studies have examined racial variation in antiresorptive medication use, those studies are limited to generalizing beyond the use of hormone therapy (HT), geographic region, and black ethnicity. Our primary aims were (1) to describe variation in the use of antiresorptive medications among US women of different ethnicities aged 45 years and older in the year 2000 and (2) to understand the association between ethnicity and antiresorptive medication use after adjustment for other factors that might have had an impact on use. The data source used in this study was the 2000 Medical Expenditure Panel Survey. Using a modified health services utilization model we performed hierarchical multivariate logistic regression analyses to examine the relationship between race and antiresorptive medication use. The use of HT among blacks, Hispanics, and other ethnicities was significantly less than among whites. This relationship was significant, even after we had controlled for predisposing, enabling, and need characteristics. The use of newer antiresorptive medications (bisphosphonates, raloxifene, and nasal calcitonin) also was less common among racial minorities than among whites. However, controlling for predisposing characteristics of education, income, census region, and age eliminated differences in use between Hispanics and whites. Controlling for need characteristics of health status, osteoporosis, and predisposing illnesses eliminated differences in antiresorptive use between blacks and whites. Significant differences in the use of newer antiresorptives remained between other ethnicities and whites, even in the fully specified model. The results of this study suggest that ethnicity is a significant predictor of the use of both HT and newer antiresorptive medication. These results also suggest that a number of variables are important in explaining racial differences in antiresorptive use, particularly among users of newer antiresorptive medications. PMID- 16314983 TI - Reference values for exposure to PAH contaminants: Comparison of fish from Ohio and Mid-Atlantic streams. AB - Reference values for background exposures of benzo[a]pyrene (BAP)- and naphthalene (NAPH)-type metabolites were calculated for white sucker, northern hog sucker, and rock bass from the mid-Atlantic region based on the 90th percentile of a probability-based sample. Preliminary findings are presented for common carp. Bile was collected from fish and assayed for polycyclic aromatic hydrocarbon metabolites using the fluorescent method of Lin et al. Exposure reference values for white sucker were 0.3 microg/mg protein for BAP-type metabolites and 40 microg/mg protein for NAPH-type metabolites. Values from the Mid-Atlantic region were similar to those previously reported for the Eastern Corn Belt Plains (ECBP) ecoregion. Exposure reference values of BAP-type metabolites for rock bass were 0.3 microg/mg protein and 0.4 microg/mg protein for northern hog sucker. Exposure reference values of NAPH-type metabolites for rock bass were 30 microg/mg protein for rock bass and 50 microg/mg protein for northern hog sucker. Provisional values for common carp based on 39 observations are 0.4 microg/mg protein for BAP-type metabolites and 120 microg/mg protein for NAPH-type metabolites. Small streams exhibited the greatest range of exposures, 0.015-0.689 microg/mg protein for BAP-type metabolites and from 5.7 to 159 microg/mg protein for NAPH-type metabolites. Larger streams had 0.11-0.859 microg/mg protein for BAP-type metabolites and 10.2-286.5 microg/mg protein for NAPH-type metabolites. Exposure reference values for BAP and NAPH-type metabolites could be used as a basis of comparison of exposure to PAH contamination. PMID- 16314984 TI - Estimating the exposure of birds and mammals to pesticides in long-term risk assessments. AB - This paper reviews current EU pesticide risk assessment guidance [European Commission (2002) Guidance document on risk assessment for birds and mammals under council directive 91/414/EEC, SANCO/4145/2000EC 2002], and examines some of its assumptions and problems arising from them. Issues associated with obtaining data that adequately describes exposure over the appropriate time-scale are common to both acute and long-term risk assessments but are probably less problematic for long-term exposure. Improvements in problem formulation and ways in which temporal and spatial factors might be incorporated into long-term risk assessments are suggested. The most important temporal issue for long-term risk is how best to model the degree to which wildlife habits are predictable from day to day. In relation to spatial factors, it is suggested that long-term risk assessments could make better use of pesticide usage data that sample usage patterns throughout the UK. The usefulness of detailed simulated farming landscapes populated by wildlife represented as agent-based models, should be explored. PMID- 16314985 TI - Multidetector spiral CT renal angiography in the diagnosis of giant renal artery aneurysms. AB - A case of giant renal artery aneurysm in a 63-year-old man is presented. Renal artery aneurysms are rare (incidence < 1%) and this case is one of the largest giant renal artery aneurysms recorded in the literature. This case also demonstrates the value of multidetector spiral computed tomographic renal angiography in the diagnosis, planning, and treatment of renal aneurysms. PMID- 16314986 TI - Elevated gastric lesions: virtual gastroscopy. AB - With the development of multidetector computed tomography and the improvement in the capabilities of workstations, the use of high-quality three-dimensional reconstructions and virtual images can be applied to organs other than the colon such as the stomach. As a noninvasive technique, virtual gastroscopy represents an alternative to conventional endoscopy for the detection of elevated lesions. Findings of this technique are illustrated. PMID- 16314987 TI - Diagnosis and management of endoleaks after endovascular aneurysm repair: role of MRI. PMID- 16314988 TI - Primary sarcoma of an abdominal aortic aneurysm. AB - Primary tumors of the aorta are extremely rare and the diagnosis is made most often after surgery or autopsy. Because clinical symptoms of abdominal sarcoma are similar to those of occlusive or aneurysmal disease, aortic sarcomas are frequently mistaken for these lesions. The imaging findings are frequently nonspecific and therefore do not allow a definitive preoperative diagnosis. We report a case of an epithelioid angiosarcoma in the vessel wall of an abdominal aortic aneurysm. PMID- 16314989 TI - Lymphoepithelial cyst of the pancreas: case report with special reference to imaging--pathologic correlation. AB - We report a case of resected lymphoepithelial cyst of the pancreas with special reference to the imaging-pathologic correlation. Visualization of a multilocular cystic nature with internal heterogeneous hyperechogenicity on ultrasound, hyperdensity on precontrast computed tomography, and granular hypointensity on T2 weighted image due to abundant internal keratin substances were considered to be keys to the differential diagnosis from other cystic lesions of the pancreas. PMID- 16314990 TI - Superior mesenteric artery aneurysm stent graft. AB - Visceral artery aneurysms represent 0.1% to 0.2% of all vascular aneurysms. They are mostly asymptomatic, but rupture is associated with a high mortality rate. We present a case of an asymptomatic aneurysm of the proximal superior mesenteric artery in a 64-year-old man that was successfully treated by implantation of a covered stent graft. The use of endovascular techniques to manage visceral artery aneurysms should be considered. PMID- 16314991 TI - Post-traumatic erectile dysfunction: doppler US findings. AB - Erectile dysfunction can happen after trauma, particularly with vertebral, pelvic, or perineal injuries. Penile Doppler ultrasonographic (US) findings in these patients are various, from normal to serious arterial impairment, according to the severity and type of injury. With neurogenic causes, Doppler US findings are usually normal, but decreased flow in cavernosal arteries may also be encountered due to combined vascular injury. With arteriogenic causes, relatively or absolutely decreased peak velocity in cavernosal arteries can be encountered. Alterations of penile arterial anatomy are frequently found in these patients and can be secondary changes due to proximal arterial insufficiency. After trauma in the penis or perineum, distortion or reconstruction of vascular anatomy in addition to traumatic sequelae in the erectile tissue can be directly visualized on Doppler US. Venogenic impotence can also be a result of trauma, and Doppler US findings are the same as nontraumatic venous leak. High-flow priapism is another category of post-traumatic erectile dysfunction, which can be diagnosed conclusively by Doppler US. It can be managed by angiographic embolization, and Doppler US is useful in evaluating recurrence and erectile dysfunction after embolization. PMID- 16314992 TI - Abdominal aortic invasion by leiomyosarcoma. AB - We describe a case of retroperitoneal leiomyosarcoma with extra- and intravascular involvement. The patient presented with abdominal aortic rupture secondary to tumor invasion and extensive tumor thrombus in the inferior vena cava. The presence of a rupture of the abdominal aorta is an important radiologic diagnosis to make because early endovascular or surgical repair can prevent a catastrophic free rupture. Inferior vena cava involvement by leiomyosarcoma may be locally resected in some cases but has a poor long-term survival rate. PMID- 16314993 TI - Congenital extrahepatic portal vein aneurysm. AB - Portal vein aneurysm is a rare clinical entity, with only 41 published cases in the English-language literature. Twenty-five of them were congenital. We present the case of a 50-year-old woman who was incidentally diagnosed with a congenital extrahepatic portal vein aneurysm during an investigation for dyspepsia. Ultrasonographic features are described with correlation of computed tomographic and magnetic resonance findings. Etiology, clinical significance, and management strategies for these lesions are discussed and a review of the literature regarding this entity is presented. PMID- 16314994 TI - Laparoscopic subtotal colectomy with cecorectal anastomosis for slow-transit constipation. AB - Subtotal colectomy with cecorectal anastomosis represents an interesting alternative to total colectomy with ileorectal anastomosis. Several technical variants to the methods for performing the anastomosis between the cecum and the rectal stump after subtotal colectomy have been reported. The mechanical, antiperistaltic, end-to-end cecorectal anastomosis is safe and easy to perform. The authors aimed to assess the safety and feasibility of this technique performed laparoscopically in a series of four patients. All the procedures were completed laparoscopically. The mean time for surgery was 200 min (range, 180-220 min). There was no mortality and no postoperative complications. The mean hospital stay was 4 days (range, 3-5 days). This technique can be performed laparoscopically with all the advantages inherent to the minimally invasive approach. PMID- 16314995 TI - An alternative gallbladder extraction technique in laparoscopic cholecystectomy. AB - In this era of minimally invasive surgery, the challenge remains in finding techniques to reduce access trauma in terms of fewer and smaller size trocar ports. Our new described technique will allow a smaller subxiphoid port to be used to achieve extraction of the gallbladder without the need to change to a 5 mm laparoscope. We believe this method is easy to learn, safe and with no observable complications from our experience. PMID- 16314996 TI - Multicenter experience with the Monarc transobturator sling system to treat stress urinary incontinence. AB - This study was conducted to report on the safety and efficacy of a transobturator sling approach for treatment of urodynamic stress incontinence (USI). Women with urodynamic stress incontinence were offered a novel polypropylene mesh sling procedure, implanted beneath the lateral pubic rami rather than retropubically. Retrospective data were collected at three U.S. sites. Two hundred women with a mean age of 59 years (range 27-93) underwent the sling procedure. Mean follow-up was 21.5 weeks (range 3-43). Perioperative safety parameters are reported for all subjects and efficacy parameters for those who were followed up for 26 weeks or more. Among the subjects, 95.3% reported being continent or substantially continent (occasional leakage of small amounts, with protection not needed). Mean operative time was 13.8 min (range 7-34). Preoperatively, 75% wore pads, with 5.3% continuing to do so at 26 weeks. Pre-op urgency was reported by 62.7%, with 41.5% receiving medication. Urgency was reported postoperatively by 20.5% of patients, but only 13.6% were medicated for urgency. Urinary retention was identified in 0.9% at 26 weeks. Transobturator slings are highly effective for treatment of SUI, and may offer increased safety relative to traditional and tension-free retropubic slings. PMID- 16314997 TI - Neural consequences of somatosensory extinction: an fMRI study. AB - There are currently two main interpretations proposing mechanisms underlying tactile extinction: sensory and attention deficit hypotheses. Kinsbourne proposed an opponent processor model to support the attention deficit hypothesis. He insisted that bilateral hemispheres interact reciprocally through contralaterally oriented vectors, and in patients presenting extinction, balance is impaired, causing inattention. From Kinsbourne's point of view, extinction is not caused by sensory disturbance but inattention, therefore even in extinction patients, simultaneous bilateral stimuli should reach the bilateral primary sensory cortices (SI). Using functional magnetic resonance imaging (fMRI), tactile stimuli were administered to both hands of healthy subjects as well as a tactile extinction patient. The patient with tactile extinction extinguished right palm stimuli following simultaneous palm stimulation. During the fMRI study, we gave tactile stimuli to the right palm, the left palm, and simultaneously to both palms. In normal subjects, simultaneous bilateral stimuli activated the bilateral SI and bilateral secondary sensory cortices (SII). In the patient with right tactile extinction, simultaneous bilateral stimuli activated the bilateral SI along with the bilateral SII and right superior parietal lobule. Our study suggests that activation of SI is insufficient to engender an awareness of sensory stimuli. From the view point of Kinsbourne, stimulus driven activity in one hemisphere suppresses activity in the other hemisphere via callosal connections. Our results support the notion that an undamaged superior parietal lobule in the patient with tactile extinction suppresses the damaged parietal lobe function and causes extinction. PMID- 16314998 TI - Acute pseudobulbar palsy due to methotrexate with rapid response to intravenous immunoglobulin. PMID- 16315003 TI - Longitudinal microPET imaging of brain tumor growth with F-18-labeled RGD peptide. AB - PURPOSE: EMD 121974, a potent cyclic RGD peptide inhibitor of alphav-integrins, demonstrated effectiveness in suppressing brain tumor growth in both preclinical models and phases I/II clinical trials. The ability to non-invasively evaluate alphav-integrin expression provides a novel and unique way to better understand brain tumor angiogenesis in relationship to alphav-integrin expression, and allow for direct assessment of anti-integrin treatment efficacy. PROCEDURES: We developed a F-18-labeled RGD peptide [F-18]FB-RGD and performed serial microPET imaging scans to follow brain tumor growth and angiogenesis as a function of time in an orthotopic U87MG glioblastoma xenograft model in athymic nude mice. RESULTS: The tumor was barely visible on microPET at the size of 5 ng/ml. PMID- 16315005 TI - Evaluation of a methylenetetrahydrofolate-dehydrogenase 1958G>A polymorphism for neural tube defect risk. AB - Genetic variants of enzymes involved in the folate pathway might be expected to have an impact on neural tube defect (NTD) risk. Given its key role in folate metabolism, the methylenetetrahydrofolate dehydrogenase 1 (MTHFD1) gene could represent an attractive candidate in NTD aetiology. In this study, the impact of the MTHFD1 1958G > A polymorphism on NTD risk in the Italian population was examined both by hospital-based case-control and family-based studies. The MTHFD1 1958G > A polymorphism was genotyped in 142 NTD cases, 125 mothers, 108 fathers and 523 controls. An increased risk was found for the heterozygous 1958G/A (OR = 1.69; P = 0.04) and homozygous 1958A/A (OR = 1.91; P = 0.02) genotypes in the children. Significant association was also found when combined 1958G/A and 1958A/A genotypes of cases were compared with the 1958G/G genotype (OR = 1.76; P = 0.02). The risk of an NTD-affected pregnancy of the mothers was increased 1.67 fold (P = 0.04) only when a dominant effect (1958G/A or 1958A/A vs 1958G/G) of the 1958A allele was analysed. The combined TDT/1-TDT (Z = 2.11; P = 0.03) and FBAT (Z = 2.4; P = 0.01) demonstrated a significant excess of transmission of the 1958A allele to affected individuals. In summary, our results indicate that heterozygosity and homozygosity for the MTHFD1 1958G > A polymorphism are genetic determinants of NTD risk in the cases examined. PMID- 16315006 TI - Propagation and functional characterization of serum-free cultured porcine hepatocytes for downstream applications. AB - Hepatocyte transplantation is considered an alternative to whole organ transplantation. However, the availability of human cadaveric livers for the isolation of transplantation-quality hepatocytes is increasingly restricted. Xenogeneic porcine hepatocytes may therefore serve as an alternate cell ressource. The propagation of hepatocytes is often necessary to yield a sufficient cell number for downstream applications in xenotransplantation and in, for example, bioartificial liver support or pharmacological and toxicological studies. Our goal has been to propagate primary porcine hepatocytes in vitro and to determine the functional maintenance of the propagated cells. Porcine hepatocytes were cultured under serum-free conditions in the presence of hepatocyte growth factor and epidermal growth factor and passaged several times. The viability, proliferation and maintenance of liver-specific functions were determined as culture proceeded. Total cell number increased by 12-fold during four sequential passages, although the proliferative capacity was higher in primary cells and early passages as compared with late passages. Xenobiotics metabolism and urea synthesis gradually decreased with ongoing culture but could be restored by treatment with appropriate stimuli such, as beta-naphthoflavone and cAMP. The expression of hepatocyte-specific genes was generally lower at the beginning than at later time-points of culture of individual passages. Porcine hepatocytes can thus be propagated in vitro. The partial loss of hepatocyte function may be restored in vitro by appropriate stimuli. This may also be achieved in a recipient liver after hepatocyte transplantation provided that the proper physiological environment for the maintenance of the differentiated hepatocyte phenotype is present. PMID- 16315007 TI - Promotion of cell adherence and spreading: a novel function of RAGE, the highly selective differentiation marker of human alveolar epithelial type I cells. AB - The receptor for advanced glycation endproducts (RAGE) is expressed under pathological conditions in many tissues and has been assigned many functions. We demonstrate, in normal human lung tissue, the preferential and highly abundant expression of RAGE by quantitative polymerase chain reaction. In addition, RAGE expression, as a specific differentiation marker of alveolar epithelial type I cells (AT I cells), and its localization to the basolateral plasma membrane have been confirmed by means of newly raised monoclonal antibodies. The physiological function of RAGE on AT I cells has previously remained elusive. By using HEK293 cells transfected with cDNA encoding for full-length RAGE, we show that RAGE enhances the adherence of epithelial cells to collagen-coated surfaces and has a striking capacity for inducing cell spreading. The preferential binding of RAGE to collagen has been confirmed by assaying the binding of soluble RAGE to various substrates. RAGE might thus assist AT I cells to acquire a spreading morphology, thereby ensuring effective gas exchange and alveolar stability. PMID- 16315008 TI - Antimicrobial agents for treatment of serious infections caused by resistant Staphylococcus aureus and enterococci. AB - As clinicians increasingly contend with infections due to staphylococci or enterococci resistant to, or failing treatment with, traditional antimicrobial agents, understanding the potential roles of older as well as more recently introduced antimicrobial agents becomes important. Older agents, such as clindamycin and trimethoprim-sulfamethoxazole, have been used to treat infections due to community-acquired methicillin-resistant Staphylococcus aureus. Among the licensed agents, quinupristin-dalfopristin, linezolid, daptomycin, and tigecycline are active in vitro against most strains of methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium, but these agents differ in their approved clinical indications. New agents currently under investigation may further expand treatment options. PMID- 16315009 TI - Histoplasmosis infection in Spanish travelers to Latin America. AB - The aim of the study presented here was to assess the incidence of histoplasma infection in a cohort of 342 individuals in Spain who had traveled to Latin America for the first time. The histoplasmin skin test was positive in 20% of the travelers, and Central America posed a higher risk for infection than South America (p=0.013). Sleeping outdoors (p=0.031) and the duration of travel (p=0.016) were also identified as significant risk factors. Serological testing demonstrated poor overall sensitivity for detecting infection in the travelers, but for the symptomatic acute cases the results were improved. Histoplasmosis must be considered in patients presenting with fever (odds ratio=3.51 [1.52 8.12]) or cough (odds ratio=4.24 [1.32-13.58]) after visiting Latin America. The results of this study have public health implications and indicate the risks of acquiring histoplasmosis should be included in pre-travel counseling. PMID- 16315010 TI - Usefulness of E-test strips for testing susceptibility of Mycobacterium tuberculosis complex strains. PMID- 16315011 TI - Betaine aldehyde dehydrogenase from Pseudomonas aeruginosa: cloning, over expression in Escherichia coli, and regulation by choline and salt. AB - In the human pathogen Pseudomonas aeruginosa, betaine aldehyde dehydrogenase (BADH) may play a dual role assimilating carbon and nitrogen from choline or choline precursors--abundant at infection sites--and producing glycine betaine, which protects the bacteria against the high-osmolarity stress prevalent in the infected tissues. We cloned the P. aeruginosa BADH gene and expressed the BADH protein in Escherichia coli. The recombinant protein appears identical to its native counterpart, as judged by Western blot, N-terminal amino acid sequence, tryptophan-fluorescence emission spectra, circular-dichroism spectroscopy, size exclusion chromatography, and kinetic properties. Computational analysis indicated that the promoter sequence of the putative operon that includes the BADH gene has a consensus-binding site for the choline-sensing transcription repressor BetI, and putative boxes for ArcA and Lrp transcription factors but no known elements of response to osmotic stress. This is consistent with the strong induction of BADH expression by choline and with the lack of effect of NaCl. As there were significant amounts of BADH protein and activity in P. aeruginosa cells grown on glucose plus choline, as well as the BADH activity exhibiting tolerance to salt, it is likely that glycine betaine is synthesized in vivo and could play an important osmoprotectant role under conditions of infection. PMID- 16315012 TI - Genotype versus phenotype in the circumscription of bacterial species: the case of Pseudomonas stutzeri and Pseudomonas chloritidismutans. AB - The phenotypic characteristic of strain AW-1(T) of Pseudomonas chloritidismutans that is most relevant from the taxonomic point of view appears to be the capacity of growth under anaerobic conditions using chlorate as electron acceptor. This property is not restricted to this species only within the genus Pseudomonas, since it is also present in strains of genomovars 1 or 5, and 3 of Pseudomonas stutzeri. P. chloritidismutans has been described as a non-denitrifying species, but the isolation of variants that are able to grow anaerobically in the presence of nitrate is possible after subcultivation under selective conditions. The subdivision of P. stutzeri into a number of species on the basis of these characteristics does not help to clarify the phylogenetic relationships among the members of an otherwise coherent group of strains, and the considerations presented in this communication support the reclassification of the new species name P. chloritidismutans, which in our opinion, should be considered as a Junior name of P. stutzeri. A multilocus sequence analysis, together with a phenotypic analysis of the anaerobic oxidative metabolism, gives new insights into the phylogeny and evolution of the species. PMID- 16315013 TI - A microfluidic SELEX prototype. AB - Aptamers are nucleic acid binding species capable of recognizing a wide variety of targets ranging from small organic molecules to supramolecular structures, including organisms. They are isolated from combinatorial libraries of synthetic nucleic acid by an iterative process referred to as SELEX (Systematic Evolution of Ligands by Exponential Enrichment). Here we describe an automated microfluidic, microline-based assembly that uses LabView-controlled actuatable valves and a PCR machine, and which is capable of the selection and synthesis of an anti-lysozyme aptamer as verified by sequence analysis. The microfluidic prototype described is 1) a simple apparatus that is relatively inexpensive to assemble, making automated aptamer selection accessible to many investigators, and 2) useful for the continued "morphing" of macro-->meso-->microfabricated structures until a convergence to a few functional systems evolves and emerges, partly or completely achieving simpler, smaller and more rapid SELEX applications. PMID- 16315014 TI - The analysis of seasonal air pollution pattern with application of neural networks. AB - Air pollution monitoring includes measuring the concentrations of air contaminants such as nitrogen dioxide, sulfur dioxide, some polycyclic aromatic hydrocarbons (PAHs), suspended particulate matter (PM) and tar substances. The purpose of this study was to determine the possibility of using artificial neural networks for identification of any patterns occurring during heating and nonheating seasons. The samples included in the study were collected over a period of 5 years (1997-2001) in the area of the city of Gdansk and the levels of pollutants measured in the samples collected were used as inputs to two different types of neural networks: multilayer perceptron (MLP) and self-organizing map (SOM). The MLP was used as a tool to predict in what heating season a certain sample was collected, and the SOM was applied for mapping all samples to recognize any similarities between them. This study also presents the comparison between two projection methods--linear (principal component analysis, PCA) and nonlinear (SOM)--in extracting valuable information from multidimensional environmental data. In the research the MLP model with 13-12-1 topology was developed and successfully trained for classification of air samples from different seasons. The sensitivity analysis on the inputs to the MLP indicated benz[alpha]anthracene, benzo[alpha]pyrene, PM1, SO2, tar substances and PM10 as the most distinctive variables, while PCA pointed to PAHs and PM1. PMID- 16315015 TI - Effect of surface coverage on the conformation and mobility of C18-modified silica gels. AB - C18-modified silica gels with surface coverages of 2 to 8.2 micromol m(-2), were prepared by different synthetic pathways and characterized by Fourier Transform infrared spectroscopy (FTIR), solid-state nuclear magnetic resonance (NMR) spectroscopy, and chromatographic measurements. The effects of temperature and bonding density on the conformational order of C18-modified silica gels were studied in detail by FTIR spectroscopy. The silane functionality and degree of cross-linking of silane ligands on the silica surface were evaluated by 29Si cross-polarization magic-angle spinning (CP/MAS) NMR and the structural order and mobility of the alkyl chains were investigated by 13C CP/MAS NMR spectroscopy. CH2 symmetric and anti-symmetric stretching bands and CH2 wagging bands were used as IR probes to monitor the conformational order and flexibility of the alkyl chains in the C18 phases. Qualitative information about the conformational order was obtained from frequency shifts of the CH2 symmetric and anti-symmetric stretching bands. The relative amounts of kink/gauche-trans-gauche, double gauche, and end-gauche conformers in the alkyl chains were determined by analysis of CH2 wagging bands. These results indicate that surface coverage plays a dominant role in the conformational order of C18-modified silica gels. The FTIR and NMR data are discussed in the context of the chromatographic shape selectivity differences. PMID- 16315016 TI - Nanoscale organic and polymeric field-effect transistors as chemical sensors. AB - This article reviews recently published work concerning improved understanding of, and advancements in, organic and polymer semiconductor vapor-phase chemical sensing. Thin-film transistor sensors ranging in size from hundreds of microns down to a few nanometers are discussed, with comparisons made of sensing responses recorded at these different channel-length scales. The vapor-sensing behavior of nanoscale organic transistors is different from that of large-scale devices, because electrical transport in a nanoscale organic thin-film transistor depends on its morphological structure and interface properties (for example injection barrier) which could be modulated by delivery of analyte. Materials used in nanoscale devices, for example nanoparticles, nanotubes, and nanowires, are also briefly summarized in an attempt to introduce other relevant nano transducers. PMID- 16315017 TI - Neural driven angiogenesis by overexpression of nerve growth factor. AB - Mechanisms regulating angiogenesis are crucial in adjusting tissue perfusion on metabolic demands. We demonstrate that overexpression of nerve growth factor (NGF) in brown adipose tissue (BAT) of NGF-transgenic mice elevates both mRNA and protein levels of vascular endothelial growth factor (VEGF) and VEGF-receptors. Increased vascular permeability, leukocyte-endothelial interactions (LEI), and tissue perfusion were measured using intravital microscopy. NGF-stimulation of adipocytes and endothelial cells elevates mRNA expression of VEGF and its receptors, an effect blocked by NGF neutralizing antibodies. These data suggest an activation of angiogenesis as a result of both: stimulation of adipozytes and direct mitogenic effects on endothelial cells. The increased nerve density associated with vessels strengthened our hypothesis that tissue perfusion is regulated by neural control of vessels and that the interaction between the NGF and VEGF systems is the critical driver for the activated angiogenic process. The interaction of VEGF- and NGF-systems gives new insights into neural control of organ vascularization and perfusion. PMID- 16315018 TI - Aberrant hypermethylation of RASSF1A promoter in ovarian borderline tumors and carcinomas. AB - The newly identified 3p21.3 tumor suppressor gene RASSF1A is inactivated by hypermethylation in variable solid tumors, including those of the lung, breast, prostate, kidney, and ovary. The purpose of this study was to evaluate the methylation status of RASSF1A in various types and stages of ovarian epithelial tumors. We analyzed the DNA methylation status of ovarian tumors using methylation-specific polymerase chain reaction in 54 frozen ovarian tumor tissues and in 97 cases of archival ovarian serous epithelial tumors using a microdissection procedure. Hypermethylation statuses were examined vs clinicopathologic findings. RASSF1A promoter methylation rates in the various types of fresh ovarian tissues were as follows: serous cystadenoma (1/5), serous tumor of borderline malignancy (2/7), serous adenocarcinoma (4/10), mucinous cystadenoma (0/5), mucinous tumor of borderline malignancy (2/7), mucinous adenocarcinoma (3/6), transitional-cell carcinoma (1/3), clear-cell carcinoma (3/3), and malignant mullerian mixed tumor (3/3). In archived serous tumor tissues, RASSF1A promoter hypermethylation was detected in serous cystadenoma (1/6, 16.6%), serous tumor of borderline malignancy (20/41, 48.8%), and in serous adenocarcinoma (25/50, 50%). The status of RASSF1A hypermethylation in borderline tumors was found to correlate statistically with the presence of microinvasion (p=0.002), peritoneal implant (p<0.001), and bilaterality (p=0.019). The RASSF1A promoter hypermethylation was frequently found in borderline tumors and carcinomas, suggesting that RASSF1A promoter hypermethylation may be a useful molecular marker for the early detection of ovarian tumors. PMID- 16315019 TI - Pathological findings in a patient with Fabry disease who died after 2.5 years of enzyme replacement. AB - We describe the postmortem findings of a 47-year-old man with Fabry disease, an X linked glycolipid storage disorder, who was on enzyme replacement therapy with recombinant alpha-galactosidase A for more than 2 years. The patient had widespread atherosclerotic coronary artery disease that culminated in a massive acute myocardial infarction. Atherosclerotic lesions were seen in the right and left coronary systems, aorta, and the basilar artery. Typical Fabry cardiomyopathy and glomerular nephropathy were found. With the exception of vascular endothelial cells, extensive glycolipid storage deposits were seen in all vascular and nonvascular cells and organ systems. We conclude that, at least in this patient, repeated infusions with alpha-galactosidase A over a prolonged period did not appreciably clear storage material in cells other than vascular endothelial cells. These findings also illustrate accelerated atherosclerosis in susceptible patients with Fabry disease. PMID- 16315020 TI - Alpha-methylacyl-CoA racemase (AMACR/P504S) protein expression in urothelial carcinoma of the upper urinary tract correlates with tumour progression. AB - Alpha-methylacyl-CoA racemase (AMACR/P504S) is a useful biomarker of prostate cancer. We evaluated the expression of AMACR in upper urinary tract urothelial carcinomas with respect to associations with tumour stage, grade and metastasis free survival. A total of 268 tumours were investigated immunohistochemically using a tissue microarray technique. AMACR expression was noted in 127 of 261 (48.7%) evaluated tumours and was associated with high tumour stage [58 of 139 (41.7%) pTa/pT1 vs. 69 of 122 (56.6%) pT2-pT4, P=0.019] and high tumour grade [44 of 137 (32.1%) low vs. 83 of 124 (66.9%) high grade, P<0.001]. In addition, AMACR expression was associated with the presence of tumour necrosis (P<0.001) and marked stromal desmoplasia (P=0.0026). This correlation indicates that increased AMACR expression might be related to hypoxia-induced changes in cancer cell metabolism, such as increased dependence on fatty acid oxidation for energy generation. Progressive disease was observed in 73 of 183 (39.9%) patients with solitary invasive carcinomas and was associated with AMACR expression (P=0.017). Multivariate analysis, however, proved only pT-stage >1 (P<0.001) and high tumour grade (P<0.001) to be independent predictors of patient outcome. In conclusion, AMACR expression correlated with advanced tumour stage and grade and may serve as an additional prognostic indicator in upper urinary tract urothelial cancer. PMID- 16315021 TI - Angiolymphoid hyperplasia with eosinophilia (epithelioid haemangioma) occurring within multiple deep lymph nodes and presenting with weight loss and raised CA 125 levels. AB - We report a case of angiolymphoid hyperplasia with eosinophilia (epithelioid haemangioma) involving multiple deep mediastinal, abdominal and intramammary lymph nodes in a 52-year-old woman with weight loss and raised CA-125 levels. The unusual clinical presentation with raised CA-125 levels and its occurrence within multiple deep visceral lymph nodes has never been reported in literature. PMID- 16315022 TI - Effect of an oily calcium hydroxide suspension (Osteoinductal) on healing of intrabony periodontal defects. A pilot study in dogs. AB - The aim of the present study was to evaluate histologically in dogs the effect of treating intrabony defects with an oily calcium hydroxide suspension (OCHS). Intrabony defects were surgically created bilaterally at the distal aspects of the maxillary first premolars and at the mesial aspects of the third premolars in two mongrel dogs. Subsequently, the defects were randomly treated with (a) access flap surgery followed by the application of an OCHS or (b) access flap surgery alone. After 8 weeks of healing, the animals were killed. Dissected blocks containing the experimental specimens were fixed in formalin, decalcified in EDTA, and embedded in paraffin. The formation of new cementum and bone was assessed histomorphometrically. In the control group, healing was predominantly characterized by the formation of a long junctional epithelium along the root surface and limited periodontal regeneration at the most apical part of the defect. The OCHS-treated defects consistently revealed periodontal regeneration (i.e., new periodontal ligament, new cementum with inserting collagen fibers, and new bone). Within the limits of the present study, it can be concluded that OCHS may favor periodontal regeneration in acute-type intrabony periodontal defects. PMID- 16315023 TI - Reprint of criteria for the clinical evaluation of dental restorative materials. 1971. PMID- 16315024 TI - [The GABA(A) receptor family: possibilities for the development of better anesthetics]. AB - Clinically used anesthetics show amnestic, sedative, hypnotic and immobilizing properties. On a molecular level these drugs affect several receptors in the cell membrane of neurons. By using genetically engineered mice a linkage can now be made between actions on certain receptors and clinically desired and undesired effects. Experiments show that a certain GABA(A) receptor subtype mediates hypnosis and immobility, whereas another subtype is involved in side-effects like sedation and hypothermia. These findings form the basis for the development of new drugs, acting highly specific and with fewer side-effects. PMID- 16315025 TI - Effects of two different regimens of continuous hormone replacement therapy on endometrial histopathology and postmenopausal uterine bleeding. AB - OBJECTIVE: To compare the effects of frequently used two different regimens of combined continuous hormone replacement therapy; 0.625 mg conjugated equine estrogen (CEE) + 2.5 mg medroxyprogesterone acetate (MPA) and 1 mg 17beta estradiol (E2) + 0.5 mg norethindrone acetate (NETA), on endometrial histopathology and postmenopausal uterine bleeding. MATERIALS AND METHODS: Two hundred and forty-six outpatient subjects aged 41-57 years were enrolled in the study conducted at the menopause clinic between November 2003 and November 2004. One hundred and thirty-nine patients were assigned to receive 0.625 mg conjugated equine estrogen + 2.5 mg medroxyprogesterone acetate (CEE/MPA), whereas 107 patients were to receive 17beta estradiol + 0.5 mg norethindrone acetate (E2/NETA). Inclusion criteria of the study were: normal values of endometrial thickness at basal evaluation, women with intact uterus, at least 12 months of amenorrhea, normal vaginal smear, bilateral mammography and biochemical blood parameters. All women were questioned every 3 months for vaginal bleeding/spotting. Endometrial sampling was performed by Pipelle catheter in the 12th month of therapy. RESULTS: For the first 3 months, vaginal bleeding/spotting rate for the CEE/MPA group was 38.7%, whereas it was higher (45%) in the E2/NETA group. For the second 3-month period, vaginal bleeding/spotting frequencies were 41.1 and 37.8%, respectively. In the third 3-month period 30.6 and 29.6%, and in the fourth 3-month period, 18.5 and 12.5% of the patients reported vaginal bleeding or spotting. None of the results of endometrial sampling have shown findings of cancer histopathology. CONCLUSION: Compared to CEE/MPA regimen, E2/NETA therapy has not shown more favorable effects on postmenopausal bleeding abnormalities. Irregular endometrial proliferation was seen more with the E2/NETA regimen. PMID- 16315026 TI - WITHDRAWN: The safety of synthetic paclitaxel by intralesional delivery with OncoGeltrade mark into skin breast cancer metastases: method and results of a clinical pilot trial. AB - Ahead of Print article withdrawn by publisher PMID- 16315027 TI - Acanthopria and Mimopriella parasitoid wasps (Diapriidae) attack Cyphomyrmex fungus-growing ants (Formicidae, Attini). AB - New World diapriine wasps are abundant and diverse, but the biology of most species is unknown. We provide the first description of the biology of diapriine wasps, Acanthopria spp. and Mimopriella sp., which attack the larvae of Cyphomyrmex fungus-growing ants. In Puerto Rico, the koinobiont parasitoids Acanthopria attack Cyphomyrmex minutus, while in Panama at least four morphospecies of Acanthopria and one of Mimopriella attack Cyphomyrmex rimosus. Of the total larvae per colony, 0-100% were parasitized, and 27-70% of the colonies per population were parasitized. Parasitism rate and colony size were negatively correlated for C. rimosus but not for C. minutus. Worker ants grasped at, bit, and in some cases, killed adult wasps that emerged in artificial nests or tried to enter natural nests. Parasitoid secondary sex ratios were female biased for eclosing wasps, while field collections showed a male-biased sex ratio. Based on their abundance and success in attacking host ants, these minute wasps present excellent opportunities to explore how natural enemies impact ant colony demography and population biology. PMID- 16315028 TI - Mapping regulatory genes as candidates for cold and drought stress tolerance in barley. AB - Cereal crop yield is greatly affected in many growing areas by abiotic stresses, mainly low temperature and drought. In order to find candidates for the tolerance genes for these stresses, 13 genes encoding for transcription factors and upstream regulators were screened by amplification and SSCP on six parental genotypes of three barley mapping populations ('Nure' x 'Tremois', 'Proctor' x 'Nudinka', and 'Steptoe' x 'Morex'), and mapped as newly developed STS, SNP, and SSCP markers. A new consensus function map was then drawn using the three maps above, including 16 regulatory candidate genes (CGs). The positions of barley cold and drought tolerance quantitative trait loci (QTLs) presently described in the literature were added to the consensus map to find positional candidates from among the mapped genes. A cluster of six HvCBF genes co-mapped with the Fr-H2 cold tolerance QTL, while no QTLs for the same trait were positioned on chromosome 7H, where two putative barley regulators of CBF expression, ICE1 and FRY1, found by homology search, were mapped in this work. These observations suggest that CBF gene(s) themselves, rather than their two regulators, are at present the best candidates for cold tolerance. Four out of 12 drought tolerance QTLs of the consensus map are associated with regulatory CGs, on chromosomes 2H, 5H, and 7H, and two QTLs with effector genes, on chromosomes 5H and 6H. The results obtained could be used to guide MAS applications, allowing introduction into an ideal genotype of favourable alleles of tolerance QTLs. PMID- 16315029 TI - Induction of anti-tumor immunity by vaccination with dendritic cells pulsed with anti-CD44 IgG opsonized tumor cells. AB - Due to the pivotal role that dendritic cells (DC) play in eliciting and maintaining functional anti-tumor T cell responses, these APC have been exploited against tumors. DC express several receptors for the Fc portion of IgG (Fcgamma receptors) that mediate the internalization of antigen-IgG complexes and promote efficient MHC class I and II restricted antigen presentation. In this study, the efficacy of vaccination with DC pulsed with apoptotic B16 melanoma cells opsonized with an anti-CD44 IgG (B16-CD44) was explored. Immature bone marrow derived DC grown in vitro with IL-4 and GM-CSF were pulsed with B16-CD44. After 48 h of pulsing, maturation of DC was demonstrated by production of IL-12 and upregulation of CD80 and CD40 expression. To test the efficacy of vaccination with DC+B16-CD44, mice were vaccinated subcutaneously Lymphocytes from mice vaccinated with DC+B16-CD44 produced IFN-gamma in response to B16 melanoma lysates as well as an MHC class I restricted B16 melanoma-associated peptide, indicating B16 specific CD8 T cell activation. Upon challenge with viable B16 cells, all mice vaccinated with DC alone developed tumor compared to 40% of mice vaccinated with DC+B16-CD44; 60% of the latter mice remained tumor free for at least 8 months. In addition, established lung tumors and distant metastases were significantly reduced in mice treated with DC+B16-CD44. Lastly, delayed growth of established subcutaneous tumors was induced by combination therapy with anti-CD44 antibodies followed by DC injection. This study demonstrates the efficacy of targeting tumor antigens to DC via Fcgamma receptors. PMID- 16315030 TI - Complete remission of liver metastasis of pancreatic cancer under vaccination with a HLA-A2 restricted peptide derived from the universal tumor antigen survivin. AB - PURPOSE: As prognosis of advanced pancreatic cancer remains gloomy, novel therapeutic modalities have to be developed. Immunotherapy, which targets tumor associated antigens of tumor cells or tumor stroma, is currently under investigation. As survivin is expressed by neoplastic and tumor endothelial cells, but rarely by normal cells, this antigen appears as an intriguing target molecule. METHODS: A 72-year old patient, suffering from pancreatic cancer refractory to gemcitabine therapy, received the survivin-based peptide vaccinations consisting of 100 mug of a modified HLA-A2 restricted survivin(96 104) epitope in Montanide(R). Each visit the patient was assessed for adverse events, quality of life and immunological response. Immuno-monitoring was performed by IFN-gamma-ELISPOT analysis of peripheral blood lymphocytes. Clinical outcome was evaluated by repetitive computed tomography. RESULTS: Under vaccination with survivin peptides the patient initially underwent partial remission of liver metastasis which proceeded after 6 months into a complete remission with a duration of 8 months. Immunological monitoring revealed strong vaccine-induced immune-reactivity against survivin. Unfortunately, after the patient was weaned from vaccination in state of no evidence of disease, he developed recurrent disease. CONCLUSION: T-cell responses against survivin expressing cells of the tumor itself and tumor endothelium should impact tumor growth and metastasis. The presented patient with pancreatic cancer is the first example of a successful application of a survivin-based vaccination in the clinical setting. An ongoing phase I/II trial with HLA-A1, -A2 and -B35 restricted survivin peptides for patients with advanced cancer will provide further information towards this notion. PMID- 16315031 TI - Polymorphisms of IL-13 and IL-4-IL-13-SNPs in patients with penicillin allergies. AB - OBJECTIVE: Although IL-4 and IL-13 share many biologic activities, some are unique to IL-13. We genotype the IL-13 and IL-4-IL-13-SNPs genes for polymorphisms that could then be used to determine associations with IgE regulation as well as levels of IL-4 and IL-13. METHODS: Eight kinds of IgE specific to penicillins were determined with radioallergosorbent test (RAST) in the sera of 158 patients with penicillin allergies and 89 healthy subjects. Serum levels of IL-4 and IL-13 were measured by using enzyme-linked immunosorbent assay (ELISA). The genotyping of IL-13Arg130Gln, IL-4-IL-13-SNP3 and IL-4-IL13-SNP4 was carried out by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. RESULTS: Among patients with positive specific IgE, significant differences between the IL-4-IL-13-SNP3 and IL-4-IL-13-SNP4 genotypes were observed between patients with positive BPA and the control group (P<0.05, P<0.05). Additionally, we also found significant differences in the IL-4-IL-13 SNP4 genotype between positive and negative BPA-IgE patients (P<0.05). However, we found no significant differences in the prevalence of these polymorphisms between any of the symptomatic groups studied and the control group. Nor were there any significant differences between levels of IL-4 and IL-13 and any of genotypes. CONCLUSION: The data suggest that IL-4-IL-13-SNP genes between IL-4 and IL-13 play a role in regulation of specific IgE levels in patients with penicillin allergies. PMID- 16315032 TI - Polymorphisms of genes CYP2D6, ADRB1 and GNAS1 in pharmacokinetics and systemic effects of ophthalmic timolol. A pilot study. AB - OBJECTIVE: To test the hypotheses that (1) CYP2D6 genotype is associated with pharmacokinetics of ophthalmic timolol and (2) variation in genotypes of ADRB1 (beta(1)-adrenoceptor) and GNAS1 (alpha-subunit of G-protein) modulate heart rate (HR), and systolic (SAP) and diastolic (DAP) arterial pressure responses to timolol. METHODS: Nineteen glaucoma patients and eighteen healthy volunteers were treated with 0.5% aqueous and 0.1% hydrogel formulations of ophthalmic timolol using a randomised cross-over design. The participants conducted head-up tilt and maximum exercise test at four visits. Plasma concentration of timolol was measured twice for glaucoma patients and ten times for healthy volunteers on each visit. Also, the genotypes for CYP2D6, ADRB1 and GNAS1 were determined. RESULTS: Among healthy volunteers using aqueous timolol, poor metabolisers (PMs, n=2) of CYP2D6 had higher maximum plasma concentrations (C(max), values 2.63 and 2.94 ng/ml), longer elimination half-lives ( T(1/2), 5.49 and 6.75 h), and higher area under-curve (AUC, 19.54 and 23.25 ng.h/ml) than intermediate [IMs, n=6, mean+/-SD 1.73+/-0.59 ng/ml (not significant), 3.30+/-0.48 h, 11.32+/-3.72 ng.h/ml], extensive (EMs, n=8, 1.60+/-0.72 ng/ml, 3.24+/-1.24 h, 8.52+/-6.12 ng.h/ml) and ultra-rapid (UMs, n=2, values 1.23 and 1.67 ng/ml, 2.22 and 2.52 h, 6.16 and 6.94 ng.h/ml) metabolisers. The IMs, EMs and UMs did not differ from each other for any of the kinetic variables. Also, the elevation of HR from rest to maximum level tended to differ between PMs and IMs, and between PMs and UMs. The pharmacokinetics and pharmacodynamics between the CYP2D6 groups did not differ with statistical significance when hydrogel timolol was used. Upon head-up tilt, the Ser49 homozygotes (n=26) had higher SAP (P=0.03) and DAP (P<0.01) than the Gly carriers (n=11). The change in DAP from rest to maximum during exercise was lower (P<0.01) in subjects with CC alleles of GNAS1 (n=13) than those with at least one T allele (n=24). CONCLUSION: The CYP2D6 poor metabolisers may be more prone to systemic adverse events with aqueous timolol than extensive metabolisers. Since CYP2D6 genotyping is not routine clinical practice, using 0.1% timolol hydrogel instead of 0.5% aqueous preparation will increase patient safety. PMID- 16315033 TI - Measurement of CYP2D6 and CYP3A4 activity in vivo with dextromethorphan: sources of variability and predictors of adverse effects in 419 healthy subjects. AB - OBJECTIVE: Dextromethorphan (DEM) shares part of the adverse event profile of opioids and is widely used as a probe drug for CYP2D6 phenotyping and for the assessment of CYP2D6 activity. It has also been used to assess CYP3A4 activity. This study examined the influence of anthropometric variables, oral contraceptives, smoking habits, mu-opioid receptor and MDR1 genetic polymorphisms and components of the DEM ratios on the variability of CYP2D6 and CYP3A4 metabolic ratios and on the occurrence of adverse events following DEM administration. METHODS: This was a retrospective analysis of a database in 419 healthy subjects. CYP2D6 and CYP3A4 metabolic ratios were measured as the log of the ratios of the amount of DEM to the amount of dextrorphan (DOR) and of the amount of DEM to the amount of 3-methoxy-morphinan (MET) excreted in urine during a 12-h time period, respectively, following the oral administration of 80 mg of dextromethorphan hydrobromide. Logistic regression was performed to examine the factors associated with changes in metabolic ratios and with the occurrence of adverse events. RESULTS: The CYP2D6 metabolic ratio allowed identification of extensive and poor metabolizers of DEM. The CYP2D6 and CYP3A4 metabolic ratios were not strictly independent one from each other. Based on multivariate analysis, the CYP2D6 metabolic ratio was a stronger independent predictor of adverse events (p<0.0001) than the CYP2D6 phenotype (p=0.05). Anthropometric variables, oral contraceptives, smoking habits, mu-opioid receptor and MDR1 genetic polymorphisms did not significantly contribute to changes in metabolic ratios or to the occurrence of adverse events. CONCLUSIONS: Dextromethorphan can be used for CYP2D6 phenotyping, but the CYP2D6 and CYP3A4 metabolic ratios are not strictly independent one from each other. The CYP2D6 metabolic ratio predicts adverse events to DEM as does CYP2D6 phenotype, and extensive metabolizer subjects are not protected against adverse events. PMID- 16315034 TI - Somatic embryogenesis and plant regeneration from different wild diploid cotton (Gossypium) species. AB - Calli were successfully induced from hypocotyls of eight wild diploid cotton species (Gossypium) on MSB (MS salts and B(5) vitamins) medium supplemented with 0.09 microM 2,4-D (2,4-dichlorophenoxyacetic acid) and 2.32 microM KT (kinetin). Plant growth regulator (PGR) combinations, adding GA(3) (Gibberellic acid), high inorganic salt stress, and PGR-free media were used to induce embryogenic calli from nonembryogenic calli. Embryogenic cultures were induced from G. aridum S. (D(4) genome), G. davidsonii K. (D(3)-d genome), G. klotzschianum A. (D(3)-k genome), G. raimondii U. (D(5) genome), and G. stocksii M. (E(1) genome). We then observed somatic embryogenesis in the five species while calli of G. africanum V. (A(1)-2 genome), G. anomalum W. (B(1) genome), and G. bickii P. (G genome) remained nonembryogenic. Somatic embryogenesis was adjusted by changing sugar sources, regulating combinations of PGRs, and using cell suspension culture. Embryos at various developmental stages produced mature and germinating embryos when cultured on filter paper placed on the media containing different sugar sources. The utility of different sugar sources promoted globular embryos developing into cotyledonary stage and increased the frequency of cotyledonary embryos developing into normal plants. Normal plantlets were regenerated from G. davidsonii, G. klotzschianum, G. raimondii, and G. stocksii. Only abnormal plantlets were obtained in G. aridum. This work will contribute to broadening the number of regenerable cotton species and provide foundations for somatic hybridization in cotton to create new germplasm. PMID- 16315036 TI - Malrotation volvulus in a neonate: a novel surgical approach. AB - A newborn presented with bilious vomiting secondary to a malrotation and presumed perinatal-onset volvulus. Laparotomy was performed at 20 h of age at which the volvulus was derotated. Nevertheless the small but not the large bowel appeared to be completely ischaemic and non-viable. A second-look laparotomy was performed 24 h later with no significant improvement. In a new approach to the problem, nothing further was done and the child was left alone on parenteral nutrition and naso-gastric aspiration for 11 weeks. Further exploration showed that 40 cm of small bowel had survived intact and restorative surgery was carried out. Parenteral nutrition was discontinued after 9 months and the child, now aged 3.5 years, is thriving. PMID- 16315035 TI - Molecular control of stem cell maintenance in shoot apical meristem. AB - Sustained post-embryonic organ initiation and development in plants depends on coordinating the formation and differentiation of pluripotent stem cells in apical meristems. Transcriptional regulation and intercellular signalling appear to play key roles in this coordination process. Here we discuss the current knowledge about the molecular regulation of stem cell maintenance in the shoot apical meristem and recent attempts to delineate the molecular signatures of "stemness" in flowering plants. We also outline contemporary molecular approaches for deciphering the process of stem cell renewal in the shoot apical meristem. PMID- 16315037 TI - Bone morphogenetic protein expression patterns in human esophageal atresia with tracheoesophageal fistula. AB - The organogenesis of esophageal atresia with tracheoesophageal fistula (EA/TEF) remains unknown. The fistula tract appears to develop from a non-branching trifurcation of the embryonic lung bud. The non-branching growth of the fistula differs from the other lung buds and suggests a deficiency in bone morphogenetic protein (BMP) signaling, since BMPs are critical to proper lung development and branching. With IRB approval, portions of newborn human proximal esophageal pouch and distal fistula samples were recovered at the time of surgical repair of EA/TEF. The tissues were processed for immunohistochemistry. Commercially available fetal tissues were used as controls. In control tissues, BMP ligands (BMP 2, 4, and 7) were all present in the esophagus but absent in the trachea. BMPRIA was absent in both tissues. BMPRIB was detected in trachea but not in esophagus and BMPRII was detected in esophagus but not in trachea. In the EA/TEF specimens, all BMP ligands were present in the proximal esophageal pouch but absent in the fistula tract. BMPRIA and BMPRIB were not detected in either tissue. However, BMPRII was found in both fistula tract and proximal pouch. The submucosa of the fistula appears to maintain a mixed (identical neither to lung, esophagus, or trachea) BMP signaling pattern, providing one mechanism which could potentially explain the esophageal dismotility and lack of lung branching seen in the fistula/distal esophagus. PMID- 16315038 TI - Management of anorectal malformations in Varanasi, India: a long-term review of single and three stage procedures. AB - To compare the results of the staged procedure (Group A) with the primary single stage posterior sagittal anorectoplasty (PSARP) procedure (Group B) for high and intermediate anorectal anomalies. Four hundred ninety cases of primary single stage procedure done over a period of seven years (1996-2003) were compared with 458 out of 763 cases of staged procedure (Historical control) that underwent all three stages done from 1989 to 1996. Two hundred cases that had been in regular follow up in Group A and 254 cases in Group B who had attained three years of age were evaluated for continence by Kelly's method. The approximate cost of treatment was also calculated. Continence was good in 90 cases (45%), fair in 66 cases (33%) and poor in 44 cases (22%) in Group A while it was good in 204 cases (80% ), fair in 38 cases (15%) and poor in 12 cases (5%) in Group B. In Group A, the mortality was high as only 40% cases completed all the three stages of operation. The mortality was 4.5% in Group B. Primary PSARP is recommended as the better treatment option for intermediate and high-anorectal anomalies with superior results in terms of morbidity, mortality, continence and cost. PMID- 16315039 TI - Ganglion of the groin in a child. PMID- 16315040 TI - HIV-1 GP120 V3 conformational and informational entropies. AB - In an attempt to analyze structure, function and evolution of HIV-1 GP120 V3, interactions among the Hartree-Fock energy, the conformational entropy and the Shannon entropy were determined for the 1NJ0 set of antibody-bound V3 loop conformers. The Hartree-Fock energy of each conformer was determined at the MINI level with GAMESS. The conformational entropy was determined per conformer and per residue from the mass-weighted covariance matrices. The Shannon entropy per residue was determined from sequence-substitution frequencies. Correlations were determined by linear regression analysis. There was a negative correlation between the Hartree-Fock energy and the conformational entropy (R=-0.4840, p=0.0078, df =28) that enhanced the negative Helmholtz-free-energy change for the binding of the GP120 ligand to target CD4. The Shannon entropy of V3 was a function of the conformational entropy variance (R=0.7225, p=0.00157, df=15) and of the V3 Hartree-Fock energy. Biological implications of this work are that (1) conformational entropy interacts with V3 Hartree-Fock energy to enhance GP120 binding to CD4 cell receptors and that (2) the Hartree-Fock energy of V3 interacts with the evolutionary system to participate in the regulation of V3 diversity. PMID- 16315041 TI - Ectopia lentis et pupillae: report of a unilateral case and surgical management. AB - PURPOSE: To report a case of clinically unilateral congenital ectopia lentis et pupillae (ELeP) and discuss its surgical management. DESIGN: Interventional case report. METHODS AND RESULTS: A 37-year-old female was referred to our service for surgical management of unilateral ectopia pupillae for aesthetic reasons. On presentation the visual acuity of the affected eye was 20/400. Ultrasound exam did not show any crystalline lens dislocation, and therefore the case was considered a very rare variable of the ELeP syndrome. We proceeded to surgical correction of the pupillary ectopia. At the 3-month follow-up the patient's visual acuity was 20/30. CONCLUSIONS: Although the clinical features and the genetic aspects of ELeP are documented in the older ophthalmic literature, this entity has received relatively little attention. Surgical correction of this rare disorder can be attempted for aesthetic and functional reasons. PMID- 16315042 TI - A familial case of pigmented paravenous retinochoroidal atrophy with asymmetrical fundus manifestations. AB - PURPOSE: To report two Japanese sibling cases with pigmented paravenous retinochoroidal atrophy (PPRCA) with asymmetrical manifestation. METHODS: A 33 year-old female and her 31-year-old brother were referred to our hospital. Funduscopic examination, angiography, and electroretinogram were performed. RESULTS: Both had retinal pigment epithelium atrophy with pigmentation along the retinal veins, which was consistent with PPRCA. The younger brother was more severely affected than his sister. Surprisingly, the extent and degree of the affected region was clearly different between both eyes of each patient. CONCLUSIONS: These findings have rarely been reported previously, and these cases stress the importance of considering gender, familial involvement, and variable manifestations of the disease in clinical care of PPRCA. PMID- 16315043 TI - Decreased blood flow at neuroretinal rim of optic nerve head corresponds with visual field deficit in eyes with normal tension glaucoma. AB - PURPOSE: To determine the relationship between the blood flow parameters of the optic disc rim and the glaucomatous visual field changes. DESIGN: Observational cross-sectional study. METHODS: Tissue blood flow in the neuroretinal rim within the optic disc was determined with the Heidelberg retina flowmeter(HRF) in 54 eyes of 54 patients with normal tension glaucoma (NTG). Patients were selected whose visual field defects were confined to either the superior or inferior hemifield. Blood flow measurements were made in a 10 degrees x 2.5 degrees area of the superior and inferior neuroretinal rim within the optic disc. The mean blood flow (MBF) was calculated by the automatic full-field perfusion image analyzer program, and the ratio of the MBF in the superior to the inferior rim areas (the S/I ratio) was calculated from the same HRF image in order to minimize the variation of measurement condition. RESULTS: Inferior rim blood flow is less than superior rim blood flow in patients with superior hemifield defect, and superior rim blood flow is reduced compared to inferior in patients with inferior hemifield defect. The mean S/I ratios of the MBF in the patients with superior hemifield defect (1.46, n=37) was significantly higher than that in the patients with inferior hemifield defect (0.79, n=17; P<0.0001, Mann-Whitney U-test). CONCLUSIONS: The blood flow in the neuroretinal rim was found to correspond to the regional visual field defect in eyes with NTG. Reductions in flow were associated with reductions in function. PMID- 16315044 TI - Acute zonal occult outer retinopathy in the fellow eye 5 years after presentation of punctate inner choroidopathy. AB - BACKGROUND: There has been argument about the relationship between posterior inner choroidopathy (PIC) and acute zonal occult outer retinopathy (AZOOR). METHOD: Case study. RESULTS: A 26-year-old woman was diagnosed as having PIC in the right eye in 1998. Multiple yellow-white lesions in the inner choroid and retina were found in the posterior pole of her right eye and were treated with corticosteroid therapy. In 2003, she was referred to us complaining of visual field defect in the left eye. By means of electroretinographic and static perimetric analyses, she was diagnosed as having AZOOR in the left eye. CONCLUSION: This case presents indirect evidence that PIC and AZOOR may have some common etiologic or pathogenic background. PMID- 16315045 TI - Treatment of large conjunctival nevus by resection and reconstruction using amniotic membrane. AB - BACKGROUND: Nevus of the bulbar conjunctiva is a benign pigmented lesion of the ocular surface. Unless a biopsy is required for ruling out malignant melanoma, a nevus is usually excised for cosmetic reasons only. However, with large lesions involving two or more quadrants of the bulbar conjunctiva, a total resection can be difficult. In this report, we present a case of a huge nevus treated by surgical excision and amniotic membrane transplantation for reconstruction of the bulbar conjunctiva. METHODS: A 54-year-old Japanese man was referred to our clinic with suspected malignant tumor of the bulbar conjunctiva in his right eye. A large and diffuse pigmented tumor with numerous small cysts was present mainly on the upper bulbar conjunctiva. Resection of the conjunctival tumor and amniotic membrane transplantation for reconstruction of the bulbar conjunctiva were performed. RESULTS: The histopathological diagnosis was conjunctival nevus. Epithelialization of the bulbar conjunctiva over the amniotic membrane sheet was completed 4 weeks after resection. At 44-month follow-up, there was no recurrence or any postoperative complication. CONCLUSION: Surgical resection combined with reconstruction by amniotic membrane transplantation is effective for the treatment of large conjunctival nevus. PMID- 16315046 TI - Microincisional cataract surgery and Thinoptx rollable intraocular lens implantation. AB - BACKGROUND: Microincisional cataract surgery is a safe procedure with a very short learning period for an experienced cataract surgeon and rollable ultrathin intraocular lenses eliminate the need for enlargement of corneal incision. The purpose of the study was to evaluate the safety and efficacy of cataract surgery through a corneal microincision and implantation of rollable ultrathin intraocular lenses. The setting was Dokuz Eylul University Medical Faculty, Ophthalmology Department, Izmir, Turkey and SSK Okmeydani Hospital, Ophthalmology Clinic, Istanbul, Turkey. PATIENTS AND METHODS: Ninety eyes in 85 patients were operated on through clear corneal microincisions with sleeveless phacoemulsification and rollable intraocular lenses were implanted. Forty-six of the patients were men and 39 were women between the ages of 27 and 83, with a mean of 51 years. Two eyes had atrophic senile macular degeneration, 4 eyes had nonspecific retinal pigment epithelial changes with chorioretinal atrophy, and 4 patients had diabetes mellitus without retinopathy. Three eyes had posterior capsular opacifications of unknown etiology. Two eyes had primary open angle glaucoma (PAAG) with cup to disc ratios of about 0.5. Three eyes had dense nuclear sclerosis of grade 4 with very low visibility of retinal structures. Other patients had no ocular or systemic pathology other than nuclear/corticonuclear cataract of grade 2-3. Uncorrected and best spectacle corrected distance and near visual acuities, keratometric values, and refractive status were noted preoperatively and 1 week, 1 month, and 6 months postoperatively. Statistical analysis of keratometric changes between preoperative and postoperative findings was performed using the paired samples t test. RESULTS: At 6 months postoperatively, 1 patient had a best spectacle corrected visual acuity (BSCVA) of 0.2, the patient with atrophic senile macular degeneration. The rest of the eyes achieved a BSCVA of 0.63 or better. At 6 months postoperatively, 55 (61.11%) eyes had uncorrected visual acuities (UCVA) equal to or better than 0.8 and 83 (92.22%) eyes had BSCVA equal to or better than 0.8 according to the Snellen chart. The mean postoperative corneal astigmatisms at 1 week, 1 month, and 6 months were 0.69+/-0.43 D, 0.66+/-0.46 D and 0.65+/-0.48 D respectively. Statistical analysis revealed a significant change in corneal astigmatisms at the 1st week visit (p<0.05), but not at the 1st and 6th month visits (p>0.05) compared with preoperative findings. CONCLUSION: Based on the limited data in the literature and in this study, it is not possible to make concrete decisions about the benefits and disadvantages of the ThinOptx IOL for longer durations. Intraoperatively, this IOL apparently eliminates the need for enlargement of the corneal incision during implantation. However, the statistical insignificance of induced astigmatisms after microincisions and classical phacoincisions should also be taken into consideration. We conclude that ThinOptx IOL is a pioneering intraocular lens implant that will contribute to the exciting future of cataract refractive surgical procedures. However, both clinical and laboratory investigations are needed to clearly describe the long term effectiveness of this new rollable IOL. PMID- 16315047 TI - Similarity of interspike interval distributions and information gain in a stationary neuronal firing. AB - The Kullback-Leibler (KL) information distance is proposed for judging similarity between two different interspike interval (ISI) distributions. The method is applied by a comparison of four common ISI descriptors with an exponential model which is characterized by the highest entropy. Under the condition of equal mean ISI values, the KL distance corresponds to information gain coming from the state described by the exponential distribution to the state described by the chosen ISI model. It has been shown that information can be transmitted changing neither the spike rate nor coefficient of variation (CV). Furthermore the KL distance offer an indication of the exponentiality of the chosen ISI descriptor (or data): the distance is zero if, and only if, the ISIs are distributed exponentially. Finally an application on experimental data coming from the olfactory sensory neurons of rats is shown. PMID- 16315048 TI - Hydrodynamic detection by cupulae in a lateral line canal: functional relations between physics and physiology. AB - In the present review, signal-processing capabilities of the canal lateral line organ imposed by its peripheral architecture are quantified in terms of a limited set of measurable physical parameters. It is demonstrated that cupulae in the lateral line canal organ can only partly be described as canal fluid velocity detectors. Deviation from velocity detection may result from resonance, and can be characterized by the extent to which a single dimensionless resonance number, N ( r ), exceeds 1. This number depends on four physical parameters: it is proportional to cupular size, cupular sliding stiffness and canal fluid density, and inversely proportional to the square of fluid viscosity. Situated in a canal, a cupula may benefit from its resonance by compensating for the limited frequency range of water motion that is efficiently transferred into the lateral line canal. The peripheral transfer of hydrodynamic signals, via canal and cupula, leads to a nearly constant sensitivity to outside water acceleration in a bandwidth that ranges from d.c. to a cut-off frequency of up to several hundreds of Hertz, significantly exceeding the cut-off frequency of the lateral line canal. Threshold values of hydrodynamic detection by the canal lateral line organ are derived in terms of water displacement, water velocity, water acceleration and water pressure gradients and are shown to be close to the detection limits imposed by hair cell mechano-transduction in combination with the physical constraints of peripheral lateral line signal transfer. The notion that the combination of canal- and cupular hydrodynamics effectively provides the lateral line canal organ with a constant sensitivity to water acceleration at low frequencies so that it consequently functions as a low-pass detector of pressure gradients, supports the appropriateness of describing it as a sense organ that "feels at a distance" (Dijkgraaf in Biol Rev 38:51-105, 1963). PMID- 16315049 TI - Anisotropic constitutive equations and experimental tensile behavior of brain tissue. AB - The present study deals with the experimental analysis and mechanical modeling of tensile behavior of brain soft tissue. A transversely isotropic hyperelastic model recently proposed by Meaney (2003) is adopted and mathematically studied under uniaxial loading conditions. Material parameter estimates are obtained through tensile tests on porcine brain materials accounting for regional and directional differences. Attention is focused on the short-term response. An extrapolation of tensile test data to the compression range is performed theoretically, to study the effect of the heterogeneity in the tensile/compressive response on the material parameters. Experimental and numerical results highlight the sensitivity of the adopted model to the test direction. PMID- 16315050 TI - The influence of fluid shear stress on the remodeling of the embryonic primary capillary plexus. AB - The primary capillary plexus in early yolk sacs is remodeled into matured vitelline vessels aligned in the direction of blood flow at the onset of cardiac contraction. We hypothesized that the influence of fluid shear stress on cellular behaviors may be an underlying mechanism by which some existing capillary channels remain open while others are closed during remodeling. Using a recently developed E-Tmod knock-out/lacZ knock-in mouse model, we showed that erythroblasts exhibited rheological properties similar to those of a viscous cell suspension. In contrast, the non-erythroblast (NE) cells, which attach among themselves within the yolk sac, are capable of lamellipodia extension and cell migration. Isolated NE cells in a parallel-plate flow chamber exposed to fluid shear stress, however, ceased lamellipodia extension. Such response may minimize NE cell migration into domains exposed to fluid shear stress. A two-dimensional mathematical model incorporating these cellular behaviors demonstrated that shear stress created by the blood flow initiated by the embryonic heart contraction might be needed for the remodeling of primary capillary plexus. PMID- 16315051 TI - Diagnosis and metaphylaxis of stone disease. Consensus concept of the National Working Committee on Stone Disease for the upcoming German Urolithiasis Guideline. AB - This review draws the recent state of the art in metabolic diagnosis and metaphylaxis of stone disease. It is the basis for the consensus approval with the other medical societies and institutions in Germany involved in the guideline process of the new "Urolithiasis Guideline". The German Working Committee on Stone Disease reviewed critically the current literature in the field of urolithiasis-including the existing German and EAU-Guidelines as well as the Conference Book of the First International Consultation on Stone Disease. As far as possible the references were rated according to the EBM criteria. On this basis the expert group discussed all pathways and statements regarding the management of stone disease. The present review coincides with the consented guideline draft of the German Working Committee on Stone Disease. Occurrence of stone disease in the western world increases seriously. Modern lifestyle, dietary habits and overweight-problems of the affluent societies-emerge to be the important promoters of the "stone-boom" in the new millennium. This even affects children, whose stone prevalence is otherwise significantly less than that of adults. Criteria for the high risk group of stone formers were clearly defined. A diagnostic standard is formulated for the basic and the elaborate metabolic evaluation of a stone patient. Approximately 75% of all stone patients could anticipate stone recurrence with elementary reorientation of their lifestyle and dietary habits, summarized as general metaphylaxis. About 25% of the stone formers require additional pharmacological intervention to normalize their individual biochemical risk, precisely compiled for each stone type as specific metaphylaxis. PMID- 16315052 TI - Water relations of the burrowing sandhill frog, Arenophryne rotunda (Myobatrachidae). AB - Arenophryne rotunda is a small (2-8 g) terrestrial frog that inhabits the coastal sand dunes of central Western Australia. While sand burrowing is a strategy employed by many frog species inhabiting Australia's semi-arid and arid zones, A. rotunda is unique among burrowing species because it lives independently of free water and can be found nocturnally active on the dune surface for relatively extended periods. Consequently, we examined the physiological factors that enable this unique frog to maintain water balance. A. rotunda was not found to have any special adaptation to reduce EWL (being equivalent to a free water surface) or rehydrate from water (having the lowest rehydration rate measured for 15 Western Australian frog species), but it was able to maintain water balance in sand of very low moisture (1-2%). Frogs excavated in the field were in dune sand of 4.4% moisture content, as a consequence of recent rain, which was more than adequate for these frogs to maintain water balance as reflected by their low plasma and urine osmotic concentrations. We suggest that in dry periods of the year, A. rotunda can achieve positive water balance by cutaneous water uptake by burrowing deeper into the substrate to where the percent water content is greater than 1.5%. PMID- 16315053 TI - Editorial: Evaluation of chemosensory effects due to occupational exposures. PMID- 16315054 TI - Outcome assessment: recommendations for daily practice. AB - The choice of instruments for the assessment of outcome in spinal surgery is bewildering. For day-to-day practice, however, consideration of the purpose for which information is required allows construction of simple strategies for data collection. Recommendations are made for short and convenient data sets for use in personal audit, clinical governance, benchmarking, patient selection and business planning. No simple data set can measure in detail every aspect of practice, but use of these recommendations will provide information that will be of great value to the spinal surgeon and ultimately to his patients. PMID- 16315055 TI - Outcome assessment and documentation: a friend or foe? PMID- 16315056 TI - Response coding in the Simon task. AB - Recent findings indicate that two distinct mechanisms can contribute to a Simon effect: a visuomotor information transmission on the one hand and a cognitive code interference on the other hand (see for e.g., Wiegand & Wascher, in Journal of Experimental Psychology: Human Perception and Performance 2005a). Furthermore, it was proposed that the occurrence of one or the other mechanism strongly depends on the way responses are coded. Visuomotor information transmission seems to depend on a correspondence between stimulus position and spatial anatomical status of the effector, whereas cognitive code interference is thought to be based on relative response location codes. To further test the spatial anatomic coding hypothesis, three experiments were conducted, in which the Simon effect with unimanual responses was investigated for horizontal (Experiment 1 and 2) and vertical (Experiment 3) stimulus-response (S-R) relations. Based on the finding of a decreasing effect function (indicating the presence of visuomotor information transmission) for horizontal and vertical S-R relations, it was concluded that visuomotor information transmission occurs whenever there is an overlap between the spatial stimulus feature and parameters of the motor representation of the response. Furthermore, the specific motor representation seems to be task dependent, that is, it entails those response parameters that clearly differentiate between the two response alternatives in a given task situation. PMID- 16315057 TI - Characterization of the novel HCH-degrading strain, Microbacterium sp. ITRC1. AB - A gram-positive Microbacterium sp. strain, ITRC1, that was able to degrade the persistent and toxic hexachlorocyclohexane (HCH) isomers was isolated and characterized. The ITRC1 strain has the capacity to degrade all four major isomers of HCH present in both liquid cultures and aged contaminated soil. DNA fragments corresponding to the two initial genes involved in gamma-HCH degradative pathway, encoding enzymes for gamma-pentachlorocyclohexene hydrolytic dehalogenase (linB) and a 2,5-dichloro-2,5-cyclohexadiene-1,4-diol dehydrogenase (linC), were amplified by PCR and sequenced. Their presence in the ITRC1 genomic DNA was also confirmed by Southern hybridization. Sequencing of the amplified DNA fragment revealed that the two genes present in the ITRC1 strain were homologous to those present in Sphingomonas paucimobilis UT26. Both 16S rRNA sequencing and phylogenetic analysis resulted in the identification of the bacteria as a Microbacterium sp. We assume that these HCH-degrading bacteria evolved independently but possessed genes similar to S. paucimobilis UT26. The reported results indicate that catabolic genes for gamma-HCH degradation are highly conserved in diverse genera of bacteria, including the gram-positive groups, occurring in various environmental conditions. PMID- 16315058 TI - Generalized arterial calcification of infancy: two siblings with prolonged survival. AB - In generalized arterial calcification of infancy (OMIM no. 208000), calcification of the media and proliferation of the intima lead to arterial stenoses. Most affected patients present with untreatable arterial hypertension and die within the first months of life. The disease has recently been linked to mutations in ENPP1. We report two siblings with prolonged survival, both of whom carry the compound heterozygous ENPP1 mutations c.913C>A and c.1164+2T>A. In both siblings, spontaneous regression of arterial calcifications occurred, and antihypertensive treatment could be tapered off gradually. In some patients, the natural course of GACI may be more favourable than previously assumed. PMID- 16315059 TI - Traumatic complications from placement of thoracic catheters and tubes. AB - The volume of critically ill patients requiring stabilization in emergency departments (EDs) throughout the USA has increased from 42 million per year in 1960 to over 92 million in 1990, as reported by Goldstein [Crit Care Clinics 21(1):81-89, 2005] and Rivers et al. [Curr Opin Crit Care 8(6):600-606, 2002]. With the increase in this patient population, the number of procedures, both invasive and noninvasive, performed in the ED to improve clinical outcomes has also increased. Therefore, emergency medicine physicians must add to their repertoire the ability to recognize potentially fatal traumatic complications. This review will provide readers with imaging findings of traumatic complications from placement of thoracic catheters and tubes and briefly discuss pitfalls of performing these procedures. In particular, complications arising from placement of hemodialysis catheters, central venous catheters, Swan-Ganz catheters, chest tubes, nasogastric and feeding tubes, and endotracheal tubes will be reviewed. PMID- 16315060 TI - Percutaneous treatment of osteoid osteoma by CT-guided drilling resection in pediatric patients. AB - BACKGROUND: Osteoid osteoma is a painful, benign, small osteogenic bone tumor. For a long time, surgery was the only treatment for these lesions. Different minimally invasive therapeutic techniques have been proposed. We report our experience in the treatment of osteoid osteoma by CT-guided drilling resection in pediatric patients. OBJECTIVE: To evaluate the efficacy of CT-guided percutaneous drilling resection as a minimally invasive therapy for osteoid osteoma in children. MATERIALS AND METHODS: Over a 5-year period, 18 patients (age range 6 17 years, mean age 11.6 years) with osteoid osteomas (femur, n=10; tibia, n=5; humerus, n=2; vertebral body, n=1) were treated with this technique. All procedures were performed under general anesthesia. RESULTS: All procedures were technically successful. Clinical success was achieved in 94.5% of patients (17/18). Only one patient had recurrence of symptoms 8 months after percutaneous resection and was surgically retreated. There were no complications. CONCLUSION: CT-guided percutaneous drilling resection is a safe, simple and effective minimally invasive technique for the treatment of osteoid osteoma in pediatric patients. PMID- 16315061 TI - Cell death in developing human spinal cord. AB - Cell death in the developing human spinal cord was investigated in 5-12 week human conceptuses using immunohistochemical and TUNEL methods. Expression of pro apoptotic (Fas-receptor, caspase-3) and anti-apoptotic (bcl-2) markers and marker for internucleosomal fragmentation (TUNEL) were analysed in the cranial and caudal parts of the human spinal cord. In early developmental stages (5-6 weeks) of the cranial spinal cord, bcl-2 positive cells were seen in the ventricular zone and in the roof plate, while in the caudal part they were seen surrounding the central lumen. Subsequently, bcl-2 expression appeared in the basal plates of the grey matter and in the spinal ganglia, and from the seventh week on they also appeared in the intermediate horn of the grey matter. In the fetal period, bcl-2 expression appeared in the dorsal horns of the grey matter (9 weeks) but ceased in the ventricular zone (12 weeks) . In the trunk region, TUNEL-positive cells were found in ventricular and mantle zones along the whole length of the spinal cord. Caspase-3 positive cells and Fas-receptor positive cells appeared only in the grey matter of the cranial segments (head and trunk) of the spinal cord, but they were missing in the caudal parts. Caspase-3 dependant pathway, probably activated by Fas-receptor, seems to operate only in the cranial part of the human spinal cord. In the caudal (sacrococcygeal and tail) parts, cells seem to die by caspase-3 independent pathway. The interplay of pro-apoptotic and anti-apoptotic factors may be associated with cranial spinal cord morphogenesis, adjustment of cells number and selective survival of neurons, while in the caudal regions these factors cause massive cell death associated with regression of the caudal spinal cord. PMID- 16315062 TI - The association between neurological deficit in acute ischemic stroke and mean transit time: comparison of four different perfusion MRI algorithms. AB - The purpose of our study was to identify the perfusion MRI (pMRI) algorithm which yields a volume of hypoperfused tissue that best correlates with the acute clinical deficit as quantified by the NIH Stroke Scale (NIHSS) and therefore reflects critically hypoperfused tissue. A group of 20 patients with a first acute stroke and stroke MRI within 24 h of symptom onset were retrospectively analyzed. Perfusion maps were derived using four different algorithms to estimate relative mean transit time (rMTT): (1) cerebral blood flow (CBF) arterial input function (AIF)/singular voxel decomposition (SVD); (2) area peak; (3) time to peak (TTP); and (4) first moment method. Lesion volumes based on five different MTT thresholds relative to contralateral brain were compared with each other and correlated with NIHSS score. The first moment method had the highest correlation with NIHSS (r = 0.79, P < 0.001) followed by the AIF/SVD method, both of which did not differ significantly from each other with regard to lesion volumes. TTP and area peak derived both volumes, which correlated poorly or only moderately with NIHSS scores. Data from our pilot study suggest that the first moment and the AIF/SVD method have advantages over the other algorithms in identifying the pMRI lesion volume that best reflects clinical severity. At present there seems to be no need for extensive postprocessing and arbitrarily defined delay thresholds in pMRI as the simple qualitative approach with a first moment algorithm is equally accurate. Larger sample sizes which allow comparison between imaging and clinical outcomes are needed to refine the choice of best perfusion parameter in pMRI. PMID- 16315063 TI - Phylogenetic relationship of the populations within and around Japan using 105 short tandem repeat polymorphic loci. AB - We have analyzed 105 autosomal polymorphic short tandem repeat (STR) loci for nine East and South-eastern Asian populations (two Japanese, five Han Chinese, Thai, and Burmese populations) and a Caucasian population using a multiplex PCR typing system. All the STR loci are genomewide tetranucleotide repeat markers of which the total number of observed alleles and the observed heterozygosity were 756 and 0.743, respectively, for Japanese populations. Phylogenetic analysis for these allele frequency data suggested that the Japanese populations are more closely related with southern Chinese populations than central and/or northern ones. STRUCTURE program analysis revealed the almost clearly divided and accountable population structure at K=2-6, that the two Japanese populations always formed one group separated from the other populations and never belong to different groups at K> or =3. Furthermore, our new allele frequency data for 91 loci were analyzed with those for 52 worldwide populations published by previous studies. Phylogenetic and multidimensional scaling (MDS) analyses indicated that Asian populations with large population size (six Han Chinese, three Japanese, two Southeast Asia) formed one distinct cluster and are closer to each other than other ethnic minorities in east and Southeast Asia. This pattern may be the caviar of comparing populations with greatly differing population sizes when STR loci were analyzed. PMID- 16315064 TI - [Kidney transplantation in childhood and adolescence]. AB - The reasons for end-stage renal disease in pediatric patients differ from adults. The therapy of choice is renal transplantation. A total of 117 children and adolescents were treated with renal transplantation in 2003 in Germany. Immunosuppressive therapy and related comorbidities are the main problems in pediatric patients. The following article provides a summary of transplantation in children, preparation, and follow-up. PMID- 16315065 TI - Elevated plasma concentrations of transforming growth factor-beta 1 in patients with unilateral ureteral obstruction. AB - We measured plasma concentrations of TGF-beta 1 in patients with obstructive ureteral calculi and compared them with the plasma concentrations of healthy volunteers. The present study was a prospective study containing a homogenous group of patients with unilateral ureteral obstruction (UUO). The study consisted of patients with ureteral stones less than 7 mm in diameter that caused mild to moderate obstruction. All patients were referred by the emergency department of our hospital and examined between April 2003 and April 2004. The presence and characteristics of both stone and obstruction were determined by plain abdominal x-ray and gray-scale ultrasonography (US). Blood samples were collected from both patients and control individuals on admission and 1 week after conservative follow-up. The plasma TGF-beta 1 concentration was determined using a quantitative sandwich enzyme immunoassay specific for TGF-beta 1. There were 35 patients with 20 women and 15 men (average age 26.8+/-5.9 years), and 15 volunteers in the control group, with nine women and six men (average age 24.2+/ 4.5 years). Average stone size was 5.6 mm+/-1.2 mm (range 3.5-7) for the patient group. US showed the presence of mild hydronephrosis in 24 and moderate hydronephrosis in 11 patients. Plasma concentrations of TGF-beta 1 in patients with ureteral obstruction (1,117+/-5.8 ng/ml, range 36-2,442 ng/ml) were significantly higher than those in the healthy control group (32+/-4 ng/ml) on admission (P<0.001). There was a significant increase in TGF-beta 1 plasma concentrations in the patient group (33,525+/-6.8 ng/ml, range 1,107-73,288 ng/ml) after 1 week follow-up (P<0.001). Ureteral obstruction increases plasma TGF-beta 1 concentrations in patients with ureteral stones as in UUO models in animal studies. A concomitant treatment with an anti-fibrotic agent may reduce the incidence of renal injury during obstruction. PMID- 16315066 TI - Topical use of tea tree oil reduces the dermal absorption of benzoic acid and methiocarb. AB - Tea tree oil (TTO) is a complex mixture of terpene hydrocarbons. Intensive topical use of TTO in different cosmetics and investigations into its potential as an antimicrobial or anti-inflammatory agent has accentuated the need for studies on the toxicity of TTO. We have applied an experimental in vitro model using static diffusion cells with human skin to study penetration characteristics of terpinen-4-ol and the way TTO affects the barrier integrity of the skin and the percutaneous penetration of two chemicals covering a range of solubilities from 0.03 g/l (methiocarb) to 3.0 g/l (benzoic acid). Through GC-MS analysis we identified the major constituents of TTO. In our experimental set-up with full thickness skin, only the least lipophilic ingredients of TTO penetrated the skin. Barrier integrity was evaluated through measurement of percutaneous penetration of tritiated water. Data indicate that 1% TTO does not affect barrier conditions. The Kp value for tritiated water was increased significantly at 5% TTO, which demonstrate that the barrier integrity is affected at this relatively low concentration of TTO. The barrier integrity is, however, not seriously damaged, but our data indicate an initiated and concentration-dependent effect on the barrier integrity. TTO changed the penetration characteristics for benzoic acid as well as for methiocarb. The general effect was that TTO reduced the maximal flux. For methiocarb, the lag-time was also prolonged by increasing the TTO concentration in the donor phase to 5%. Thus, TTO reduced the overall amount of benzoic acid as well as methiocarb entering the receptor chamber. PMID- 16315067 TI - [Differential diagnosis of bone marrow edema of the knee joint]. AB - Bone marrow edema of the knee joint is a frequent clinical picture in MR diagnostics. It can be accompanied by symptoms and pain in the joint. Diseases that are associated with bone marrow edema can be classified into different groups. Group 1 includes vascular ischemic bone marrow edema with osteonecrosis (synonyms: SONK or Ahlback's disease), osteochondrosis dissecans, and bone marrow edema syndrome. Group 2 comprises traumatic or mechanical bone marrow edema. Group 3 encompasses reactive bone marrow edemas such as those occurring in gonarthrosis, postoperative bone marrow edemas, and reactive edemas in tumors or tumor-like diseases. Evidence for bone marrow edema is effectively provided by MRI, but purely morphological MR information is often unspecific so that anamnestic and clinical details are necessary in most cases for definitive disease classification. PMID- 16315068 TI - [Muscle and tendon injuries of the knee joint]. AB - Muscles and tendons contribute greatly to stabilization of the knee joint and are crucial elements for normal joint function. Therefore, injuries of those structures cause variable degrees of disability, depending on the severity of the lesion. Due to the characteristic structural changes and alterations of signal intensities, MR imaging allows accurate identification and staging of acute injuries of muscles and tendons, as well as assessment of chronic or degenerative disease. The knowledge of anatomy and normal function of muscles and tendons, together with a thorough understanding of the mechanism and usual combinations of their injuries (e.g., posterolateral structures) allows an exact evaluation of injury-induced functional impairments to the knee joint. A detailed and exact radiological report is of the greatest importance for the referring clinician and the patient to determine optimal therapeutic strategies. PMID- 16315069 TI - The chemosensory basis for behavioral divergence involved in sympatric host shifts. I. Characterizing olfactory receptor neuron classes responding to key host volatiles. AB - The recent shift of Rhagoletis pomonella from its native host hawthorn to introduced, domestic apple has been implicated as an example of sympatric speciation. Recent studies suggest that host volatile preference might play a fundamental role in host shifts and subsequent speciation in this group. Single sensillum electrophysiology was used to test a proposed hypothesis that differences in R. pomonella olfactory preference are due to changes in the number or odor specificity of olfactory receptor neurons. Individuals were analyzed from apple, hawthorn, and flowering dogwood-origin populations, as well as from the blueberry maggot, Rhagoletis mendax Curran (an outgroup). Eleven compounds were selected as biologically relevant stimuli from previous electroantennographic/behavioral studies of the three R. pomonella populations to host fruit volatiles. Cluster analysis of 99 neuron responses showed that cells from all tested populations could be grouped into the same five classes, ranging from those responding to one or two volatiles to those responding to several host volatiles. Topographical mapping also indicated that antennal neuron locations did not differ by class or fly taxa. Our results do not support the hypothesis that differences in host preference among Rhagoletis populations are a result of alterations in the number or class of receptor neurons responding to host volatiles. PMID- 16315070 TI - The chemosensory basis for behavioral divergence involved in sympatric host shifts II: olfactory receptor neuron sensitivity and temporal firing pattern to individual key host volatiles. AB - The Rhagoletis species complex has been a key player in the sympatric speciation debate for much of the last 50 years. Studies indicate that differences in olfactory preference for host fruit volatiles could be important in reproductively isolating flies infesting each type of fruit via premating barriers to gene flow. Single sensillum electrophysiology was used to compare the response characteristics of olfactory receptor neurons from apple, hawthorn, and flowering dogwood-origin populations of R. pomonella, as well as from the blueberry maggot, R. mendax (an outgroup). Eleven volatiles were selected as stimuli from behavioral/electroantennographic studies of the three R. pomonella host populations. Previously, we reported that differences in preference for host fruit volatile blends are not a function of alterations in the general class of receptor neurons tuned to key host volatiles. In the present study, population comparisons involving dose-response trials with the key volatiles revealed significant variability in olfactory receptor neuron sensitivity and temporal firing pattern both within and among Rhagoletis populations. It is concluded that such variability in peripheral sensitivity and temporal firing pattern could influence host preference and contribute to host fidelity and sympatric host shifts in the Rhagoletis complex. PMID- 16315071 TI - Ganglioneuromas of the sacrum-a report of two cases with radiologic-pathologic correlation. AB - Ganglioneuromas are rare, benign, well-differentiated, slow-growing tumors, composed of ganglion cells and Schwann cells. Ganglioneuromas are derived from the neural crest cells and can arise anywhere from the base of the skull to the pelvis. We present and discuss the clinicopathologic and radiographic features of two patients with ganglioneuroma arising from the sacrum, a rare anatomic location. PMID- 16315072 TI - Rapid detection and high-resolution discrimination of the genus Streptomyces based on 16S-23S rDNA spacer region and denaturing gradient gel electrophoresis. AB - As the leading source of antibiotics, Streptomyces species are the subject of widespread investigation. Many approaches have been tried to aid in the classification of Streptomyces isolates to the genus, species, and strain levels. Genetic methods are more rapid and convenient than classification methods based on phenotypic characteristics, but a method that is universal in detecting all Streptomyces yet selective in detecting only Streptomyces is needed. The highly conserved nature of the 16S rRNA gene (16S rDNA) combined with the need to discriminate between closely related strains results in analyses of ribosomal intergenic spacer (RIS) regions being more productive than analyses of 16S rRNA genes. PCR primers were designed to amplify the RIS region as well as a sufficient length of the 16S rRNA gene to enable phylogenetic analyses of Streptomyces. Improved selectivity and specificity for the amplification of RIS sequences from Streptomyces with environmental samples was demonstrated. The use of RIS-PCR and denaturing gradient gel electrophoresis (DGGE) was shown to be a convenient means to obtain unique genetic "fingerprints" of Streptomyces cultures allowing them to be accurately identified at species, and even strain classification levels. These RIS-PCR and DGGE approaches show potential for the rapid characterization of environmental Streptomyces populations. PMID- 16315073 TI - The 2004 WHO classification of pituitary tumors: comments. PMID- 16315074 TI - [Computer assisted chewing power in patients with segmental resection of the mandible]. AB - BACKGROUND: Surgical treatment of tumors of the oral cavity often requires a segmental resection of the mandible. This always implies a considerable loss in function and aesthetics. The aim of the present study was to measure the chewing force obtained by patients after mandibular resection. PATIENTS AND METHODS: In a group of 20 patients (twelve males, eight females, average age 59 years), chewing force were registered by means of a computerized measurement device. In 16 patients, the defect was bridged by a reconstruction plate, in three with an iliac bone graft stabilized by miniplates, and in one patient with only two miniplates. RESULTS: The maximum value in the molar region was 186 N und the minimum was 28 N. The average bite force in the molar region reached 81.1 N (+/ 46.1) with 42.9 N (+/-35.7) in the front region. The highest value was registered in a patient with an iliac bone graft without soft tissue defect. The lowest was found in patients with bony chin defects. CONCLUSION: Based on these results, a reduction of 76% in the molar region and 59% in the incisor region was observed. These values might be helpful in providing a more realistic definition of the functional loadings found in patients after mandibular resection, which in turn may help in the development of new reconstruction devices. PMID- 16315076 TI - Redescription of Houssayela sudobim (Woodland, 1935) (Cestoda: Proteocephalidea), a parasite of Pseudoplatystoma fasciatum (Pisces: Siluriformes) from the River Amazon. AB - The proteocephalidean cestode Houssayela sudobim (Woodland, 1935), the type and only species of Houssayela Rego, 1987, is redescribed on the basis of re examination of its type-specimens and freshly collected material found in the intestine of the barred sudobim Pseudoplatystoma fasciatum (Siluriformes: Pimelodidae) from the River Amazon in Peru. The validity of the genus is confirmed, but its diagnosis is amended because suckers possess four pairs of cone-shaped (papilla-like) projections on their anterior margins (rather than 16, as reported in the original description), the actual number of testes is more than 400 in a proglottis (not 150 as given by Woodland, 1935) and the vitelline follicles form one dorsolateral medullary band and one lateral cortical band on both sides of proglottides. In addition, the distal (terminal) portion of the vagina has a double sphincter and the cirrus-sac has very thick walls; neither of these characteristics was mentioned in the original description. The specimen collected in Peru represents the first record of the species since its original description in 1935. PMID- 16315075 TI - Glutathione homeostasis and redox-regulation by sulfhydryl groups. AB - Continuous control of metabolism and developmental processes is a key feature of live cells. Cysteine thiol residues of proteins are both exceptionally useful in terms of structural and regulatory aspects, but at the same time exceptionally vulnerable to oxidation. Conserved cysteines thus are highly important for the function of metabolic enzymes and for signaling processes underlying responses to environmental factors. The underlying mechanism for the central role of thiol mediated redox control in cellular metabolism is the ability of the cysteine thiols to reversibly change their redox state followed by changes of structural, catalytic or regulatory functions. The cellular glutathione/glutathione disulfide redox buffer is present in cells at millimolar concentrations and forms one major basis of redox homeostasis by which protein thiols can maintain their redox state or oxidized protein thiols can be reverted to their reduced state. Besides acting as redox buffer, glutathione also acts as an electron donor for both scavenging of reactive oxygen, e.g. from photosynthesis and respiration, and metabolic reactions such as reduction of hydroperoxides and lipidperoxides or sulfate assimilation. The central role of glutathione is further emphasized by its involvement in signaling processes and the crosstalk of redox signaling processes with other means of signaling including protein glutathionylation and control of transcription factors. The present review aims at highlighting the key functions of glutathione in thiol-mediated redox control and its interplay with other protein-thiol-based redox systems. PMID- 16315077 TI - Description of adult and fourth-stage larva of Litomosoides navonae n. sp. (Nematoda: Onchocercidae), a parasite of five species of sigmodontine rodents from northeastern Argentina. AB - This study describes a new species of Litomosoides Chandler, 1931, parasitic in five different sigmodontine rodents from Misiones, Chaco and Formosa provinces of Argentina. The fourth-stage (L4) larva (male and female) is also described. L. navonae n. sp. exhibits: a bottle-shaped buccal cavity; a buccal capsule with irregularly crenate external walls; four externo-labial papillae and one ventral cephalic papilla; a well differentiated oesophagus; and sigmodontis-type spicules. The microfilaria is fusiform, with a large sheath. The L4 has a buccal capsule which is relatively longer than that of the adults, with narrower walls and a bottle-shaped lumen. It was observed in this species that the oesophagus/body-length ratio increases from larva to adult (female ratio 26.2 28.3 in larva; 88.4 in adults), and the vulva appears to move further posterior to the oesophago-intestinal junction (200-300 microm in larvae vs a mean of 600 microm in adults). L. navonae was found parasitising: Nectomys squamipes from the Reserve UNLP Valle del Arroyo Cuna Piru, Misiones; Oligoryzomys chacoensis, Holochilus chacarius and Akodon azarae bibianae from the marshes of Arroyo Bellaco, El Colorado, Formosa; and O. fornesi and H. chacarius from Selvas del Rio de Oro (Chaco). Both N. squamipes and H. chacarius harbour other filarioids species, i.e. L. kohnae Bain, Petit & Diagne, 1989 and L. patersoni (Mazza, 1928), respectively, throughout their range, but these filarioids are readily differentiated from L. navonae. These well-differentiated filarial species found in Nectomys and Holochilus could indicate how isolated the populations of rodents are and could be interpreted either as: (a) an early point in the speciation processes that could be taking place in these hosts; or (b) extra support for the capture phenomenon theory of the evolution of Litomosoides. New regions, such as southern Brazil and northern Argentina, need to be studied in order to clarify these alternatives. PMID- 16315078 TI - Gorgoderina festoni n. sp. (Digenea: Gorgoderidae) in Anurans (Amphibia) from Mexico. AB - Gorgoderina festoni n. sp. is described from the urinary bladder of Gastrophryne usta, Leptodactylus labialis, L. melanonotus and Bufo marinus from localities at low altitude in the states of Veracruz, Oaxaca, Guerrero and Colima, Mexico. This species differs from most other species of the genus by a combination of the following characters: lobed vitelline masses, body size 3.45-4.26 (mean 3.75) mm and sucker-ratio 1:1.3-1.52 (mean 1:1.44). The new species shares these three features with G. bilobata Rankin, 1937, G. schistorchis Steelman, 1938, G. tenua Rankin, 1937, G. vitelliloba (Olsson, 1876) and G. cryptorchis Travassos, 1924, but it differs from the first four in having gonads with entire margins. G. festoni most closely resembles G. cryptorchis, but differs from this species in body width at the level of the ventral sucker and in the absence of oesophageal glands. The autapomorphy that distinguishes G. festoni from all other members of the genus is the presence of a tegumental extension (festoon) on the external margin of the ventral sucker. PMID- 16315079 TI - Redescription of Cetorhinicola acanthocapax Beveridge & Campbell, 1988 (Cestoda: Trypanorhyncha) from the basking shark Cetorhinus maximus (Gunnerus). AB - Cetorhinicola acanthocapax Beveridge & Campbell, 1988 is redescribed based on adult specimens collected from a basking shark Cetorhinus maximus (Gunnerus) off the east coast of New Zealand. Both the mature and gravid segments are described for the first time, and the first scanning electron micrographs of the armature and scolex microtriches are provided. Novel features include the opening of the vagina anterior to the cirrus-sac and the extension of the vagina anterior to the cirrus-sac. Pectinate microtriches are present on the anterior quarter of the pars pedunculus scolecis scattered between filiform microtriches; the bothrial margins are covered with tridigitate microtriches, while the adherent surface of the bothria have pectinate microtriches. The additional morphological features described are consistent with a close association between Cetorhinicola Beveridge & Campbell, 1988 and the Eutetrarhynchoidea Guiart, 1927. PMID- 16315080 TI - Trematodes from Red Sea fishes: Neohypocreadium aegyptense n. sp. (Lepocreadiidae), Fairfaxia cribbi n. sp. and Macvicaria chrysophrys (Nagaty & Abdel-Aal, 1969) (Opecoelidae). AB - Specimens of the marine fishes Chaetodon lineolatus (Chaetodontidae), Lethrinus nebulosus (Lethrinidae) and Acanthopagrus bifasciatus (Sparidae) were caught in the Red Sea off the coast of Sharm El-Sheikh, South Sinai, Egypt. Fifteen (75%), four (16%) and fourteen (35%) fish, respectively, were found to harbour intestinal trematodes. C. lineolatus was parasitised by Neohypocreadium aegyptense n. sp. (Lepocreadiidae), L. nebulosus by Fairfaxia cribbi n. sp. (Opecoelidae) and A. bifasciatus by Macvicaria chrysophrys (Nagaty & Abdel-Aal, 1969) Bray, 1985 (Opecoelidae). N. aegyptense n. sp. is most similar to N. chaetodoni (Mahavi, 1972), but is smaller and differs in having acinous rather than digitate ovarian lobes, vitelline follicles extending anteriorly to midway between the ventral sucker and the intestinal bifurcation and an external seminal vesicle extending posteriorly to reach the anterior margin of the ovary. The generic diagnosis of Neohypocreadium is amended. F. cribbi n. sp. resembles F. lethini Cribb, 1990, but differs in having relatively smaller gonads, cirrus-sac and eggs, and larger suckers and pharynx. M. chrysophrys, collected from its type host and locality, is redescribed. Plagioporus saoudi Ramadan, 1985 is considered its synonym. PMID- 16315081 TI - Morphological characterisation of Andrya Railliet, 1893, Neandrya n. g. and Paranoplocephala Luhe, 1910 (Cestoda: Anoplocephalidae) in rodents and lagomorphs. AB - The taxonomic significance of the main morphological features of the 25 species allocated to Andrya Railliet, 1893 and Paranoplocephala Luhe, 1910 is re evaluated in the light of the recent molecular phylogenetic hypotheses for anoplocephaline cestodes. The present analysis and the existing phylogenetic data suggest that the structure and complexity of the early uterus are not, as previously assumed, the main phylogenetic or systematic determinants for anoplocephaline cestodes. Instead, the position of the early uterus with respect to other organs, combined with the morphology of the female genitalia, appear to allow a fairly straightforward discrimination of the three genera recognised here, without contradicting current phylogenetic hypotheses. A new genus, Neandrya n. g., is proposed for N. cuniculi (Blanchard, 1891) n. comb. (previously in Andrya), amended diagnoses are provided for Andrya and Paranoplocephala and a diagnostic key to these three genera is presented. PMID- 16315082 TI - Pseudorhabdosynochus summanoides n. sp. (Monogenea: Diplectanidae) from Epinephelus coioides in Dapeng Bay, South China Sea, with observations on several similar species of Pseudorhabdosynochus Yamaguti, 1958. AB - One new species of diplectanid monogenean, Pseudorhabdosynochus summanoides n. sp., is reported and described from the marine fish Epinephelus coioides off Nan'ao Shenzhen, China. P. summanoides differs from its closest relative, P. summanae (Young, 1968), by the shape of its vaginal hard-parts, which have a tightly twisted distal region and an accessory patch on the proximal curve. During the course of this work, type-material of several species of Pseudorhabdosynochus was examined and aspects of the vagina and haptor are redescribed and/or figured. These species include P. americanus (Price, 1937), P. hargisi (Oliver & Paperna, 1984), P. amplidiscatus (Bravo-Hollis, 1954), P. epinepheli (Yamaguti, 1938), P. riouxi (Oliver 1986), P. melanesiensis (Laird, 1958), P. cupatus (Young, 1969), P. bocquetae (Oliver & Paperna, 1984), P. kritskyi Dyer et al., 1995, P. capurroi Vidal-Martinez, 1998, P. querni (Yamaguti, 1968) and P. summanae (Young, 1969). Several closely related species are considered in terms of their conspecificity: P. hargisi is proposed as a junior synonym of P. americanus; P. capurroi is suggested as a likely synonym of P. kritskyi; and it is suggested that P. cupatus and P. bocquetae may eventually be demonstrated to be consepcific with P. melanesiensis. PMID- 16315085 TI - The first international conference on transgenic rabbits. PMID- 16315086 TI - A French academic network for sharing transgenic materials and knowledge. PMID- 16315087 TI - Studying human pathogens in animal models: fine tuning the humanized mouse. AB - Humanized mice are crucial tools for studying human pathogens in systemic situations. An animal model of human coronavirus infectious disease has been generated by gene transfer of the human receptor for virus-cell interaction (aminopeptidase N, APN, CD13) into mice. We showed that in vitro and in vivo infections across the species barrier differ in their requirements. Transgenic cells were susceptible to human coronavirus HCoV-229E infection demonstrating the requirement of hAPN for viral cell entry. Transgenic mice, however, could not be infected suggesting additional requirements for in vivo virus susceptibility. Crossing hAPN transgenic mice with interferon unresponsive Stat1(-/- )mice resulted in markedly enhanced virus replication in vitro but did not result in detectable virus replication in vivo. Adaptation of the human virus to murine cells led to successful infection of the humanized transgenic mice. Future genetic engineering approaches are suggested to provide animal models for the better understanding of human infectious diseases. PMID- 16315088 TI - Evaluating the induced-odour emission of a Bt maize and its attractiveness to parasitic wasps. AB - The current discussion on the safety of transgenic crops includes their effects on beneficial insects, such as parasitoids and predators of pest insects. One important plant trait to consider in this context is the emission of volatiles in response to herbivory. Natural enemies use the odours that result from these emissions as cues to locate their herbivorous prey and any significant change in these plant-provided signals may disrupt their search efficiency. There is a need for practical and reliable methods to evaluate transgenic crops for this and other important plant traits. Moreover, it is imperative that such evaluations are done in the context of variability for these traits among conventional genotypes of a crop. For maize and the induction of volatile emissions by caterpillar feeding this variability is known and realistic comparisons can therefore be made. Here we used a six-arm olfactometer that permits the simultaneous collection of volatiles emitted by multiple plants and testing of their attractiveness to insects. With this apparatus we measured the induced odour emissions of Bt maize (Bt11, N4640Bt) and its near-isogenic line (N4640) and the attractiveness of these odours to Cotesia marginiventris and Microplitis rufiventris, two important larval parasitoids of common lepidopteran pests. Both parasitoid species were strongly attracted to induced maize odour and neither wasp distinguished between the odours of the transgenic and the isogenic line. Also wasps that had previously experienced one of the odours during a successful oviposition divided their choices equally between the two odours. However, chemical analyses of collected odours revealed significant quantitative differences. The same 11 compounds dominated the blends of both genotypes, but the isogenic line released a larger amount of most of these. These differences may be due to altered resource allocation in the transgenic line, but it had no measurable effect on the wasps' behaviour. All compounds identified here had been previously reported for maize and the differential quantities in which they were released fall well within the range of variability observed for other maize genotypes. PMID- 16315089 TI - Qualitative and quantitative PCR methods for event-specific detection of genetically modified cotton Mon1445 and Mon531. AB - Based on the DNA sequences of the junctions between recombinant and cotton genomic DNA of the two genetically modified (GM) cotton varieties, herbicide tolerance Mon1445 and insect-resistant Mon531, event-specific primers and probes for qualitative and quantitative PCR detection for both GM cotton varieties were designed, and corresponding detection methods were developed. In qualitative PCR detection, the simplex and multiplex PCR detection systems were established and employed to identify Mon1445 and Mon531 from other GM cottons and crops. The limits of detection (LODs) of the simplex PCR were 0.05% for both Mon1445 and Mon531 using 100 ng DNA templates in one reaction, and the LOD of multiplex PCR analysis was 0.1%. For further quantitative detection using TaqMan real-time PCR systems for Mon1445 and Mon531, one plasmid pMD-ECS, used as reference molecule was constructed, which contained the quantitative amplified fragments of Mon1445, Mon531, and cotton endogenous reference gene. The limits of quantification (LOQs) of Mon1445 and Mon531 event-specific PCR systems using plasmid pMD-ECS as reference molecule were 10 copies, and the quantification range was from 0.03 to 100% in 100 ng of the DNA template for one reaction. Thereafter, five mixed cotton samples containing 0, 0.5, 0.9, 3 and 5% Mon1445 or Mon531 were quantified using established real-time PCR systems to evaluate the accuracy and precision of the developed real-time PCR detection systems. The accuracy expressed as bias varied from 1.33 to 8.89% for tested Mon1445 cotton samples, and from 2.67 to 6.80% for Mon531. The precision expressed as relative standard deviations (RSD) were different from 1.13 to 30.00% for Mon1445 cotton, and from 1.27 to 24.68% for Mon531. The range of RSD was similar to other laboratory results (25%). Concluded from above results, we believed that the established event-specific qualitative and quantitative PCR systems for Mon1445 and Mon531 in this study are acceptable and suitable for GM cotton identification and quantification. PMID- 16315090 TI - Production of two vaccinating recombinant rotavirus proteins in the milk of transgenic rabbits. AB - Rotaviruses are the main cause of infantile viral gastroenteritis worldwide leading to approximately 500,000 deaths each year mostly in the developing world. For unknown reasons, live attenuated viruses used in classical vaccine strategies were shown to be responsible for intussusception (a bowel obstruction). New strategies allowing production of safe recombinant non-replicating rotavirus candidate vaccine are thus clearly needed. In this study we utilized transgenic rabbit milk as a source of rotavirus antigens. Individual transgenic rabbit lines were able to produce several hundreds of micrograms per ml of secreted recombinant VP2 and VP6 proteins in their milk. Viral proteins expressed in our model were immunogenic and were shown to induce a significant reduction in viral antigen shedding after challenge with virulent rotavirus in the adult mouse model. To our knowledge, this is the first report of transgenic mammal bioreactors allowing the rapid co-production of two recombinant viral proteins in milk to be used as a vaccine. PMID- 16315091 TI - The effect of DNA repair defects on reproductive performance in nucleotide excision repair (NER) mouse models: an epidemiological approach. AB - In this study, we used an epidemiological approach to analyze an animal database of DNA repair deficient mice on reproductive performance in five Nucleotide Excision Repair (NER) mutant mouse models on a C57BL/6 genetic background, namely CSA, CSB, XPA, XPC [models for the human DNA repair disorders Cockayne Syndrome (CS) and xeroderma pigmentosum (XP), respectively] and mHR23B (not associated with human disease). This approach allowed us to detect and quantify reproductive effects based on a relatively small number of matings. We measured and quantified the scale of the effect between factors that might influence reproductive performance (i.e. age at co-housing, seasons) and reproductive parameters (i.e. litter size and pairing-to-birth interval -'pbi'). Besides, we detected and quantified the differences in reproductive performance between wild type mice and heterozygous/homozygous NER mutant mice. From our analyses, we found impaired reproduction in heterozygous and homozygous knock out mice; in particular, reduced litter size and lengthened pbi was related to the NER mutation-mHR23B, in heterozygous couples, even if they were otherwise phenotypically normal. Heterozygous mHR23B couples produced a 6.6-fold lower number of mHR23B(-/-) pups than indicated by Mendelian expectation; other genetic deficiencies studied were not statistically significant from each other or wild type controls. We concluded that careful epidemiological evaluations by analysis of animal database could provide reliable information on reproductive performance and detect deviations that would remain unnoticed without this. Also, some managerial aspects of mouse breeding could be evaluated. PMID- 16315092 TI - A comparative risk assessment of genetically engineered, mutagenic, and conventional wheat production systems. AB - Wheat (Triticum aestivum L.) varieties produced using modern biotechnologies, such as genetic engineering and mutagenic techniques, have lagged behind other crop species, but are now being developed and, in the case of mutagenic wheat, commercially grown around the world. Because these wheat varieties have emerged recently, there is a unique opportunity to assess comparatively the potential environmental risks (human health, ecological, and livestock risks) associated with genetically engineered, mutagenic, and conventional wheat production systems. Replacement of traditional herbicides with glyphosate in a glyphosate tolerant (genetically engineered) wheat system or imazamox in an imidazolinone tolerant (mutagenic) wheat system may alter environmental risks associated with weed management. Additionally, because both systems rely on plants that express novel proteins, the proteins and plants themselves may impose risks. The purpose of our study was to examine comparatively the multiple aspects of risk associated with different wheat production systems in the US and Canada using the risk assessment paradigm. Specifically, we used tier 1 quantitative and qualitative risk assessment methods to compare specific environmental risks associated with the different wheat production systems. Both glyphosate and imazamox present lower human health and ecological risks than many other herbicides associated with conventional wheat production systems evaluated in this study. The differences in risks were most pronounced when comparing glyphosate and imazamox to herbicides currently with substantial market share. Current weight-of-evidence suggests that the transgenic CP4 EPSPS protein present in glyphosate-tolerant wheat poses negligible risk to humans, livestock, and wildlife. Risk for mutated AHAS protein in imidazolinone-tolerant wheat most likely would be low, but there are not sufficient effect and exposure data to adequately characterize risk. Environmental risks for herbicides were more amenable to quantitative assessments than for the transgenic CP4 EPSPS protein and the mutated AHAS protein. PMID- 16315093 TI - Expression of functional interleukin-12 from mouse in transgenic tomato plants. AB - Transgenic plants have been employed successfully as a low-cost system for the production of therapeutically valuable proteins, including antibodies, antigens and hormones. Here, we report the expression of a cytokine with immunomodulatory function, mouse interleukin-12 (IL-12), in transgenic tomato plants. Single-chain mouse IL-12 driven by the CaMV 35S promoter, accumulates to high levels in leaves and fruits (up to 7.3 and 3.4 microg per gram of fresh weight, respectively). Mouse IL-12 expressed in tomato displays biological activity in vitro, as determined by interferon-gamma (IFN-gamma) secretion by T cells. Possible uses of this plant-based cytokine involving mucosal delivery are discussed. PMID- 16315094 TI - A microarray-based comparative analysis of gene expression profiles during grain development in transgenic and wild type wheat. AB - Global, comparative gene expression analysis is potentially a very powerful tool in the safety assessment of transgenic plants since it allows for the detection of differences in gene expression patterns between a transgenic line and the mother variety. In the present study, we compared the gene expression profile in developing seeds of wild type wheat and wheat transformed for endosperm-specific expression of an Aspergillus fumigatus phytase. High-level expression of the phytase gene was ensured by codon modification towards the prevalent codon usage of wheat genes and by using the wheat 1DX5HMW glutenin promoter for driving transgene expression. A 9K wheat unigene cDNA microarray was produced from cDNA libraries prepared mainly from developing wheat seed. The arrays were hybridised to flourescently labelled cDNA prepared from developing seeds of the transgenic wheat line and the mother variety, Bobwhite, at three developmental stages. Comparisons and statistical analyses of the gene expression profiles of the transgenic line vs. that of the mother line revealed only slight differences at the three developmental stages. In the few cases where differential expression was indicated by the statistical analysis it was primarily genes that were strongly expressed over a shorter interval of seed development such as genes encoding storage proteins. Accordingly, we interpret these differences in gene expression levels to result from minor asynchrony in seed development between the transgenic line and the mother line. In support of this, real time PCR validation of results from selected genes at the late developmental stage could not confirm differential expression of these genes. We conclude that the expression of the codon-modified A. fumigatus phytase gene in the wheat seed had no significant effects on the overall gene expression patterns in the developing seed. PMID- 16315095 TI - Analysis of substrate specificity of pig CYP2B22 and CYP2C49 towards herbicides by transgenic rice plants. AB - We introduced two novel types of pig (Sus scrofa) cytochrome P450, CYP2B22 and CYP2C49, into rice plants (Oryza sativa L. cv. 'Nipponbare') to produce herbicide tolerant plants and to confirm the metabolic activities of the cytochrome P450 species. In germination tests, both types of transgenic plants showed tolerance to various herbicides with different modes of action. CYP2B22 rice plants showed tolerance towards 12 herbicides including chlortoluron (100 microM), amiprofos methyl (2.5 microM), pendimethalin (10 microM), metolachlor (2.5 microM), and esprocarb (20 microM). CYP2C49 rice plants showed tolerance towards 13 herbicides, including chlortoluron (100 microM), norflurazon (0.5 microM), amiprofos-methyl (2.5 microM), alachlor (0.8 microM), and isoxaben (1 microM). The herbicide tolerance was considered to reflect the substrate specificity of the introduced P450 species. We used (14)C-labeled metolachlor and norflurazon to confirm the P450 activity in the transgenic rice plants. The herbicides were metabolized more quickly in the transgenic rice plants than in the nontransgenic rice plants. Therefore, CYP2B22 and CYP2C49 rice plants became more tolerant to various herbicides than nontransgenic control plants because of accelerated metabolism of the herbicides by the introduced P450 species. Assuming that public and commercial acceptance is forthcoming, these transgenic rice plants may become useful tools for the breeding of herbicide-tolerant crops. PMID- 16315096 TI - The serine/threonine kinase, Krct, affects endbud morphogenesis during murine mammary gland development. AB - STK16/Krct (Kinase related to cerevisiae and thaliana) is a ubiquitously expressed member of a unique family of serine/threonine protein kinases that is conserved among all eukaryotes. Despite its cloning 6 years ago to date, the function of this kinase remains unknown. In an attempt to identify a function for Krct, we have generated a doxycycline-dependent transgenic mouse model that permits the inducible overexpression of Krct in the mammary glands of mice treated with tetracycline derivatives. Analysis of these mice reveals that modest overexpression of Krct in the mammary gland during puberty results in duplication of the terminal endbud axis such that multiple, rather than single, budding structures arise at the ends of primary ducts. Supernumerary endbuds in Krct overexpressing mice resemble wild-type terminal endbuds with regard to cellular proliferation rates and localization of cap cells, myoepithelial cells and body cells. However, aberrant transgenic endbuds are surrounded by an increased amount of periductal stroma that in many cases encompasses the entire endbud. These data suggest that Krct may play a role in regulating stromal-epithelial interactions that occur during ductal morphogenesis in the mammary gland. PMID- 16315097 TI - Characterization of a strong, constitutive mung bean (Vigna radiata L.) promoter with a complex mode of regulation in planta. AB - We report the cloning and characterization in tobacco and Arabidopsis of a Vigna radiata L. (mung bean) promoter that controls the expression of VR-ACS1, an auxin inducible ACC synthase gene. The VR-ACS1 promoter exhibits a very unusual behavior when studied in plants different from its original host, mung bean. GUS and luciferase in situ assays of transgenic plants containing VR-ACS1 promoter fusions show strong constitutive reporter gene expression throughout tobacco and Arabidopsis development. In vitro quantitative analyses show that transgenic plants harboring VR-ACS1 promoter-reporter constructs have on average 4-6 fold higher protein and activity levels of both reporter genes than plants transformed with comparable CaMV 35S promoter fusions. Similar transcript levels are present in VR-ACS1 and CaMV 35S promoter lines, suggesting that the high levels of gene product observed for the VR-ACS1 promoter are the combined result of transcriptional and translational activation. All tested deletion constructs retaining the core promoter region can drive strong constitutive promoter activity in transgenic plants. This is in contrast to mung bean, where expression of the native VR-ACS1 gene is almost undetectable in plants grown under normal conditions, but is rapidly and highly induced by a variety of stimuli. The constitutive behavior of the VR-ACS1 promoter in heterologous hosts is surprising, suggesting that the control mechanisms active in mung bean are impaired in tobacco and Arabidopsis. The 'aberrant' behavior of the VR-ACS1 promoter is further emphasized by its failure to respond to auxin and cycloheximide in heterologous hosts. VR-ACS1 promoter regulatory mechanisms seem to be different from all previously characterized auxin-inducible promoters. PMID- 16315098 TI - Development of novel types of plastid transformation vectors and evaluation of factors controlling expression. AB - Two new vector types for plastid transformation were developed and uidA reporter gene expression was compared to standard transformation vectors. The first vector type does not contain any plastid promoter, instead it relies on extension of existing plastid operons and was therefore named "operon-extension" vector. When a strongly expressed plastid operon like psbA was extended by the reporter gene with this vector type, the expression level was superior to that of a standard vector under control of the 16S rRNA promoter. Different insertion sites, promoters and 5'-UTRs were analysed for their effect on reporter gene expression with standard and operon-extension vectors. The 5'-UTR of phage 7 gene 10 in combination with a modified N-terminus was found to yield the highest expression levels. Expression levels were also strongly dependent on external factors like plant or leaf age or light intensity. In the second vector type, named "split" plastid transformation vector, modules of the expression cassette were distributed on two separate vectors. Upon co-transformation of plastids with these vectors, the complete expression cassette became inserted into the plastome. This result can be explained by successive co-integration of the split vectors and final loop-out recombination of the duplicated sequences. The split vector concept was validated with different vector pairs. PMID- 16315099 TI - Pax1/E2a double-mutant mice develop non-lethal neural tube defects that resemble human malformations. AB - Many mouse models exist for neural tube defects (NTDs), but only few of them are relevant for human patients that are born alive with spina bifida aperta. NTDs in humans show a complex inheritance, which most likely result from the involvement of a variety of predisposing genetic and environmental factors. Hints toward the identity of predisposing genetic factors for human NTDs could come from mouse studies on the development of the neural tube and spinal cord, as well as from studies on associated features of this type of diseases. Among such features is the observation that pregnancies affected by a neural tube defect frequently show changes in thymus morphology, and in both neonatal and maternal T-cell repertoire. The genes for E2a and Pax1 have both been implicated in not only paraxial mesodermal development, but also in that of the immune system. Moreover, Pax1 mutant mice have been shown to display NTDs in digenic mouse models. In the present study we have investigated the phenotype of E2a null mutant mice that are also heterozygous for the so-called undulated mutation in Pax1. Here we report that such double-mutant mice develop a non-lethal NTD that strongly resembles the classic human NTD: spina bifida aperta, associated with defects of the axial skeleton, immune system and urinary tract. PMID- 16315100 TI - Hairpin RNA-mediated silencing of Plum pox virus P1 and HC-Pro genes for efficient and predictable resistance to the virus. AB - We report the application of the hairpin-mediated RNA silencing technology for obtaining resistance to Plum pox virus (PPV) infection in Nicotiana benthamiana plants. Four sequences, covering the P1 and silencing suppressor HC-Pro genes of an Italian PPV M isolate, were introduced into N. benthamiana plants as two inverted repeats separated by an intron sequence under the transcriptional control of the Cauliflower Mosaic Virus 35S promoter. In a leaf disk infection assay, 38 out of 40 T0 transgenic plants were resistant to PPV infection. Eight lines, 2 for each construct, randomly selected among the 38 resistant plants were further analysed. Two hundred forty eight out of 253 T1 transgenic plants were resistant to local and systemic PPV infection. All transgenic single locus lines were completely resistant. These data indicate that the RNA silencing of PPV P1/HCPro sequences results in an efficient and predictable PPV resistance, which may be utilized in obtaining stone fruit plants resistant to the devastating Sharka disease. PMID- 16315101 TI - Iron status indicators in women with prior neural tube defect-affected pregnancies. AB - OBJECTIVES: Iron deficiency is the most common nutrient deficiency in pregnant women and has been linked to negative impacts on the fetus. We describe the association of various iron-deficiency indices with risk of neural tube defect (NTD) among a high-risk Mexican-American population. METHODS: The study included 158 case-women (NTD-affected pregnancies) and 189 control-women (normal births) who were residents of the 14 Texas-Mexico border counties and delivered or terminated pregnancies during 1995-2000. In-person interviews and laboratory assays provided data. RESULTS: Case-women had higher odds of having minimal or no iron stores (serum ferritin <30 microg/L) compared to control-women (OR = 1.8, 95% CI = 1.0-3.3). The risk effect was not explained by low folate or B12 or other risk factors. CONCLUSION: Low serum ferritin may reflect the additive effect of multiple long-term factors, many of them related to poverty such as poor quality diet, lack of supplementation, and frequent pregnancies. Interpartum care is indicated in this population. PMID- 16315102 TI - New fluorescent labels: 4- and 7-chlorofluorescein. AB - A new mixture of 4- and 7-chlorofluorescein were synthesized by condensation of resorcinol with 3-chlorophthalic anhydride in the presence of methanesulfonic acid or zinc chloride. These regioisomers were successfully separated by chromatography. The photophysical properties were examined and their absorption and emission maxima at long wavelength, high fluorescence quantum yield, and narrow emission bandwidth were found, highly favorable for detecting multiple target substances in the same sample. Furthermore, 4(7)-chlorofluorescein was found to be strongly pH-dependent between 4.0 and 8.0, and could be used as pH sensitive fluorescent probe to measure intracellular pH. PMID- 16315103 TI - High probe intensity photobleaching measurement of lateral diffusion in cell membranes. AB - Lateral diffusion measurements, most commonly accomplished through Fluorescence Photobleaching Recovery (FPR or FRAP), provide important information on cell membrane molecules' size, environment and participation in intermolecular interactions. However, serious difficulties arise when these techniques are applied to weakly expressed proteins of either of two types: fusions of membrane receptors with visible fluorescent proteins or membrane molecules on autofluorescent cells. To achieve adequate sensitivity in these cases, techniques such as interference fringe FPR are needed. However, in such measurements, cytoplasmic species contribute to the fluorescence recovery signal and thus yield diffusion parameters not properly representing the small number of surface molecules. A new method helps eliminate these difficulties. High Probe Intensity (HPI)-FPR measurements retain the intrinsic confocality of spot measurements to eliminate interference from fluorescent cytoplasmic species. However, HPI-FPR methods lift the previous requirement that FPR procedures be performed at probe beam intensities low enough to not induce bleaching in samples during measurements. The high probe intensities now employed provide much larger fluorescence signals and thus more information on molecular diffusion from each measurement. We report successful measurement of membrane dynamics by this technique. PMID- 16315104 TI - Nonhomologous recombination between the large unassigned region of the male and female mitochondrial genomes in the mussel, Mytilus trossulus. AB - Doubly uniparental inheritance of mtDNA (DUI) is commonly observed in several genera of bivalves. Under DUI, female offspring inherit mtDNA from their mothers, while male offspring inherit mtDNA from both parents but preferentially transmit the paternally inherited mtDNA to their sons. Several studies have shown that the female- and male-specific mtDNA lineages in blue mussels, Mytilus spp., vary by upward of 20% at the nucleotide level. In addition to high levels of nucleotide substitution, the present study observed substantial gender-based length polymorphism in the presumptive mitochondrial control region (=large unassigned region; LUR) of North American M. trossulus. In this species, female lineage LUR haplotypes are over 2 kb larger than male lineage LUR haplotypes. Analysis of sequence data for these length variants indicates that the F LUR haplotypes of North American M. trossulus contain sequences similar to the F lineage control region in the congeners M. edulis and M. galloprovincialis. Relative to the F LUR in the latter two species, however, the F lineage LUR haplotypes in M. trossulus contain two large sequence insertions, each nearly 1 kb in size. One of these insertions has high sequence similarity to the male lineage LUR of M. trossulus. The tandem arrangement of F and M control region sequences in the F lineage LUR of M. trossulus is most likely the result of nonhomologous recombination between the male and the female mitochondrial genomes in M. trossulus, a finding that has important implications regarding the transmission and evolution of blue mussel mitochondrial genomes. PMID- 16315105 TI - Transposable element orientation bias in the Drosophila melanogaster genome. AB - Nonrandom distributions of transposable elements can be generated by a variety of genomic features. Using the full D. melanogaster genome as a model, we characterize the orientations of different classes of transposable elements in relation to the directionality of genes. DNA-mediated transposable elements are more likely to be in the same orientation as neighboring genes when they occur in the nontranscribed region's that flank genes. However, RNA-mediated transposable elements located in an intron are more often oriented in the direction opposite to that of the host gene. These orientation biases are strongest for genes with highly biased codon usage, probably reflecting the ability of such loci to respond to weak positive or negative selection. The leading hypothesis for selection against transposable elements in the coding orientation proposes that transcription termination poly(A) signal motifs within retroelements interfere with normal gene transcription. However, after accounting for differences in base composition between the strands, we find no evidence for global selection against spurious transcription termination signals in introns. We therefore conclude that premature termination of host gene transcription due to the presence of poly(A) signal motifs in retroelements might only partially explain strand-specific detrimental effects in the D. melanogaster genome. PMID- 16315106 TI - Evolutionary relationships and protein domain architecture in an expanded calpain superfamily in kinetoplastid parasites. AB - Employing whole-genome analysis we have characterized a large family of genes coding for calpain-related proteins in three kinetoplastid parasites. We have defined a total of 18 calpain-like sequences in Trypanosoma brucei, 27 in Leishmania major, and 24 in Trypanosoma cruzi. Sequence characterization revealed a well-conserved protease domain in most proteins, although residues critical for catalytic activity were frequently altered. Many of the proteins contain a novel N-terminal sequence motif unique to kinetoplastids. Furthermore, 24 of the sequences contain N-terminal fatty acid acylation motifs indicating association of these proteins with intracellular membranes. This extended family of proteins also includes a group of sequences that completely lack a protease domain but is specifically related to other kinetoplastid calpain-related proteins by a highly conserved N-terminal domain and by genomic organization. All sequences lack the C terminal calmodulin-related calcium-binding domain typical of most mammalian calpains. Our analysis emphasizes the highly modular structure of calpains and calpain-like proteins, suggesting that they are involved in diverse cellular functions. The discovery of this surprisingly large family of calpain-like proteins in lower eukaryotes that combines novel and conserved sequence modules contributes to our understanding of the evolution of this abundant protein family. PMID- 16315107 TI - Gene duplication and the properties of biological networks. AB - Patterns of network connection of members of multigene families were examined for two biological networks: a genetic network from the yeast Saccharomyces cerevisiae and a protein-protein interaction network from Caenorhabditis elegans. In both networks, genes belonging to gene families represented by a single member in the genome ("singletons") were disproportionately represented among the nodes having large numbers of connections. Of 68 single-member yeast families with 25 or more network connections, 28 (44.4%) were located in duplicated genomic segments believed to have originated from an ancient polyploidization event; thus, each of these 28 loci was thus presumably duplicated along with the genomic segment to which it belongs, but one of the two duplicates has subsequently been deleted. Nodes connected to major "hubs" with a large number of connections, tended to be relatively sparsely interconnected among themselves. Furthermore, duplicated genes, even those arising from recent duplication, rarely shared many network connections, suggesting that network connections are remarkably labile over evolutionary time. These factors serve to explain well-known general properties of biological networks, including their scale-free and modular nature. PMID- 16315108 TI - Inferring the pattern of spontaneous mutation from the pattern of substitution in unitary pseudogenes of Mycobacterium leprae and a comparison of mutation patterns among distantly related organisms. AB - The pattern of spontaneous mutation can be inferred from the pattern of substitution in pseudogenes, which are known to be under very weak or no selective constraint. We modified an existing method (Gojobori T, et al., J Mol Evol 18:360, 1982) to infer the pattern of mutation in bacteria by using 569 pseudogenes from Mycobacterium leprae. In Gojobori et al.'s method, the pattern is inferred by using comparisons involving a pseudogene, a conspecific functional paralog, and an outgroup functional ortholog. Because pseudogenes in M. leprae are unitary, we replaced the missing paralogs by functional orthologs from M. tuberculosis. Functional orthologs from Streptomyces coelicolor served as outgroups. We compiled a database consisting of 69,378 inferred mutations. Transitional mutations were found to constitute more than 56% of all mutations. The transitional bias was mainly due to C-->T and G-->A, which were also the most frequent mutations on the leading strand and the only ones that were significantly more frequent than the random expectation. The least frequent mutations on the leading strand were A-->T and T-->A, each with a relative frequency of less than 3%. The mutation pattern was found to differ between the leading and the lagging strands. This asymmetry is thought to be the cause for the typical chirochoric structure of bacterial genomes. The physical distance of the pseudogene from the origin of replication (ori) was found to have almost no effect on the pattern of mutation. A surprising similarity was found between the mutation pattern in M. leprae and previously inferred patterns for such distant taxa as human and Drosophila. The mutation pattern on the leading strand of M. leprae was also found to share some common features with the pattern inferred for the heavy strand of the human mitochondrial genome. These findings indicate that taxon-specific factors may only play secondary roles in determining patterns of mutation. PMID- 16315109 TI - Evidence for Hox gene duplication in rainbow trout (Oncorhynchus mykiss): a tetraploid model species. AB - We examined the genomic organization of Hox genes in rainbow trout (Oncorhynchus mykiss), a tetraploid teleost derivative species, in order to test models of presumptive genomic duplications during vertebrate evolution. Thirteen putative clusters were localized in the current rainbow trout genetic map; however, analysis of the sequence data suggests the presence of at least 14 Hox clusters. Many duplicated genes appear to have been retained in the genome and share a high percentage of amino acid similarity with one another. We characterized two Hox genes located within the HoxCb cluster that may have been lost independently in other teleost species studied to date. Finally, we identified conserved syntenic blocks between salmonids and human, and provide data supporting two new linkage group homeologies (i.e., RT-3/16, RT-12/29) and three previously described homeologies (RT-2/9, RT-17/22, and RT-27/31) in rainbow trout. PMID- 16315110 TI - The mitochondrial genome of Xiphinema americanum sensu stricto (Nematoda: Enoplea): considerable economization in the length and structural features of encoded genes. AB - The complete sequence of the mitochondrial genome of the plant parasitic nematode Xiphinema americanum sensu stricto has been determined. At 12626bp it is the smallest metazoan mitochondrial genome reported to date. Genes are transcribed from both strands. Genes coding for 12 proteins, 2 rRNAs and 17 putative tRNAs (with the tRNA-C, I, N, S1, S2 missing) are predicted from the sequence. The arrangement of genes within the X. americanum mitochondrial genome is unique and includes gene overlaps. Comparisons with the mtDNA of other nematodes show that the small size of the X. americanum mtDNA is due to a combination of factors. The two mitochondrial rRNA genes are considerably smaller than those of other nematodes, with most of the protein encoding and tRNA genes also slightly smaller. In addition, five tRNAs genes are absent, lengthy noncoding regions are not present in the mtDNA, and several gene overlaps are present. PMID- 16315113 TI - Common injuries related to weightlifting: MR imaging perspective. AB - Weightlifting has evolved to become a ubiquitous form of exercise. Resistance training has been shown to have beneficial effects on both muscle and osseous maintenance and development. Competitive weightlifting sports continue to enjoy tremendous popularity, with participants striving to establish new standards in performance and more demanding personal goals. Thus, it is not surprising that we have also seen an increase in injuries related to weightlifting. Many of these injuries are radiographically occult and are best suited for evaluation by magnetic resonance (MR) imaging because many involve the soft tissues. In this article, we discuss some of the factors that contribute to these injuries and address the mechanisms of injury and the MR imaging manifestations of the more common injuries. PMID- 16315114 TI - Mechanisms and imaging of football-related injuries. AB - American football is a cutting/pivoting contact sport with high-speed and high energy collisions among players on every play during games and training or practice. Although there have been constant rules and equipment changes in an attempt to limit the frequency of injuries, the incidence of musculoskeletal injuries remains high at every level of play in this sport. Imaging plays an important role in managing these players by helping in making an accurate diagnosis as well as guiding therapy and monitoring response to treatment. This article describes the major mechanisms and patterns of injury seen in these athletes, with a discussion of how imaging, primarily magnetic resonance imaging, assists the team physician in choosing a conservative versus surgical approach to recovery and return to play. PMID- 16315115 TI - Imaging of the overhead throwing athlete. AB - Knowledge of overhead throwing biomechanics is crucial to understand specific injuries encountered in throwing athletes on diagnostic imaging. Most specific injuries of overhead throwing athletes occur at the shoulder and elbow. Throwing athletes are susceptible to rotator cuff tears from tensile overload and external and internal impingement. The labrum is also commonly degenerated or torn secondary to overuse syndrome, internal impingement, and microtrauma. The elbow is typically injured secondary to excessive valgus forces during throwing. The ulnar collateral ligament, ulnar nerve, and common flexor tendon origin are all at increased risk of injury. Capitellar osteochondral injuries and loose intra articular bodies are also frequent. Knowledge of injury pathophysiology is crucial to understanding the treatment rationale in throwing athletes. PMID- 16315116 TI - Imaging of rock climbing injuries. AB - Competition climbing has grown increasingly in popularity, and many people are being drawn to this sport with a parallel increase in the occurrence of sport related injuries. One of the most common and unique lesions occurring in the rock climbing population is the closed rupture of the flexor pulley system of the fingers. This lesion is strictly related to some climbing techniques in which the entire body weight is placed on fingerholds, which causes bowstringing of the flexor tendons with subsequent loss of strength across the full range of motion of the finger. This article summarizes the current literature regarding the application of imaging modalities in the diagnosis of rock climbing injuries with a specific focus on ultrasound and magnetic resonance imaging. Biomechanics of the sporting activity and resultant pathophysiologic and clinical considerations concerning flexor pulley system injuries are also discussed. PMID- 16315117 TI - Golf and racquet sports injuries. AB - There are specific injuries that are common in golf and racquet sports. These abnormalities have a predilection for specific structures as well and can be divided into two categories on the basis of etiology as either chronic repetitive injury or acute trauma. With golf injuries, upper extremity abnormalities prevail and include rotator cuff disease, epicondylitis, wrist tenosynovitis, and hamate hook fracture. Thoracolumbar spine pain can also occur. The order of frequency of these ailments is different for professional and recreational athletes. With racquet injuries, as in tennis, lower extremity injuries are more common and include medial gastrocnemius and Achilles tendon abnormalities, although shoulder, elbow, and wrist abnormalities may also occur. Knowledge of the biomechanics behind each sport is also helpful in understanding the pathophysiology of injury and in part explains the findings seen at imaging. PMID- 16315118 TI - MRI of snow skiing and snowboarding injuries. AB - Snow skiing has been called "the world's oldest sport," "the fastest non motorized sport on Earth," and "the riskiest sport undertaken by adults on a routine basis." This article discusses the common mechanisms for orthopedic injuries occurring in two of the most popular winter sports in the world that have a reputation for an inherently high risk of injury: alpine skiing and snowboarding. The emphasis herein is on magnetic resonance imaging of characteristic injuries in skiers (knee, thumb, shoulder) and in snowboarders (wrist, forearm, ankle). Spine injuries in snowboarders and skiers are also discussed. PMID- 16315119 TI - Rowing injuries. AB - The sport of rowing has gained considerable momentum in recent years. It appeals to many age groups and is an endurance activity with no sudden accelerations or ballistic impact forces. It is associated with several injuries that are so typical that they are easily recognized by the enthusiast and in many cases do not require imaging. These include wrist tenosynovitis, intersection syndrome, and rib fracture. Other injuries may be the result of strenuous training programs or faulty rowing technique and include low back injuries and patellar maltracking. The etiology, biomechanics, imaging, and treatment of rowing-related disorders are discussed. PMID- 16315120 TI - The biomechanics and imaging of soccer injuries. AB - Soccer is one of the most popular sports worldwide and requires the player to have considerable stamina while superimposing sprinting, jumping, tackling, rapid changes of direction, and kicking. This contact sport carries a significant risk for acute and overuse injuries, with the lower limb the region most frequently injured. This article discusses the biomechanics and current role of imaging in the management of several specific soccer injuries highlighting injuries of the ankle, pelvis, and groin and specific muscle and tendon injuries commonly encountered in this athletic population. PMID- 16315121 TI - [Prospects of gastroenterology]. PMID- 16315122 TI - [Future directions of German gastroenterology--personal notes on the occasion of the 60th annual meeting of the German Gastroenterological Association]. PMID- 16315123 TI - [Clinical gastroenterology--luxury or standard of service in gastroenterology?]. AB - Gastroenterology is one of the important specialities in internal medicine. The reform of the training curriculum for internal medicine and the reimbursement for inpatient and outpatient services in gastroenterology threatens the existence of internal medicine and gastroenterology in Germany, too. The capacity for training in internal medicine and gastroenterology is reduced by a decrease in the number of hospital beds in academic and community training centres. The concentration on gastrointestinal endoscopy in outpatient gastroenterology will be a result of an increasing demand for gastrointestinal endoscopy services and the decreasing number of gastroenterology clinics, respectively. Therefore, clinical gastroenterology as a core service in gastroenterology will be steadily eliminated. This development will diminish clinical gastroenterology to gastrointestinal endoscopy by eliminating the clinical services for chronic gastroenterological conditions such as, e.g., IBD, chronic hepatitis, reflux disease, IBS and functional dyspepsia. In this way gastroenterology looses its central role in health care services in specialised internal medicine. In 2003 the American Gastroenterological Association position paper: "Training the Gastroenterologist of the Future: the Gastroenterology Core Curriculum" was published. It has emphasised the role of clinical gastroenterology in medical training and medical services, too. Clinical gastroenterology consists of an array of several disciplines, e.g., GI physiology, GI research, infectious diseases, hepatology, oncology and gastrointestinal endoscopy, which all contribute to the effectiveness and efficiency in health care service. Financial incentives and better prospects of leading positions for young gastroenterologists in clinical gastroenterology have to be accomplished in order to nourish clinical gastroenterology in Germany. The German Association of Gastroenterology should negotiate with the responsible authorities for the addition of clinical gastroenterological services to the reimbursement by the EBM2000plus. The section of Gastroenterology of the German Association of Internists will provide sustained support to the achievement of this goal. PMID- 16315124 TI - Impact of etiology on the painful early stage of chronic pancreatitis: a long term prospective study. AB - BACKGROUND AND AIM: This prospective study aimed to investigate the impact of etiology on the pain profile in relation to alterations of function and morphology from early to advanced chronic pancreatitis (CP). METHODS: Our mixed medico-surgical cohort comprised 265 patients with alcoholic (ACP), 21 with idiopathic "juvenile" (IJCP), 46 with idiopathic "senile" (ISCP) and 11 with hereditary CP (HPCP). The patients were followed regularly from onset of disease according to the protocol published previously. RESULTS: Males predominated in ACP, IJCP, ISCP (> 71%) but not in HP (46%). Age at onset (median) was 10, 23, 36 and 62 years in HP, IJCP, ACP and ISCP, respectively. Follow-up from disease onset ranged from 14 to 36 years. The progression to late-stage CP, documented by exocrine insufficiency (86-100%) and calcification (80-91%) lasted 2 to 5-fold longer in HP/IJCP compared to ACP. Early stage CP, characterized by recurrent pancreatitis prevailed in > or = 90% of patients, except for those with ISCP (48%), and lasted up to 5-fold longer in HP/IJCP compared to ACP. Surgery for severe pain was required for ACP/IJCP in 57% of the patients compared to < 27% in HP/ISCP. Permanent pain relief regularly occurred in late-stage CP irrespective of etiology and surgery. CONCLUSION: The clinical profile of the 4 "etiological" subgroups is predictably different in the painful early (precalcific) CP stage. PMID- 16315125 TI - A prospective multicenter clinical and endoscopic follow-up study of patients with gastroesophageal reflux disease. AB - BACKGROUND: Despite its high prevalence, data on the course of gastroesophageal reflux disease (GERD) are still sparse. There is also an ongoing debate on the natural history of its various manifestations, i.e., non-erosive reflux disease (NERD) vs. erosive reflux disease (ERD) vs. Barrett's esophagus (BE). PATIENTS AND METHODS: This study was conducted within the framework of a prospective multicenter study on the course and prevalence of intestinal metaplasia at the gastroesophageal junction. 1014 dyspeptic patients were screened by the means of upper GI endoscopy. Clinical data were recorded by the use of a questionnaire. At least 15 months after the initial examination, all patients were invited for follow-up (FU) examination. Patients were analyzed separately with respect to their clinical and endoscopic findings. For the latter, only patients without any treatment with proton-pump-inhibitors (PPI) prior to initial and follow-up examinations were included. Patients were categorized as GERD positive if typical symptoms were present and/or proton pump inhibitors were used. RESULTS: Sufficient clinical and/or endoscopic data were available in 590 GERD patients with heartburn at least once a week. Follow-up data could be obtained in 320 patients (clinical FU: n = 304; endoscopic FU: n = 52) after a mean follow-up period of 35 months (18-48 months). 96 of 144 previously asymptomatic patients (67%) remained asymptomatic at follow-up, the rest (n = 48) were symptomatic. 143 of 304 previously symptomatic patients (47%) were symptom-free at follow-up, and only 161 patients (53%) remained symptomatic or had concomitant therapy with proton-pump-inhibitors (PPI). For follow-up endoscopy in patients off PPI (n = 52), ERD was no longer confirmed in 7/12 ERD patients (58%), whereas progress to ERD was found in 3/34 patients (9%) in the NERD group. BE was newly diagnosed in two NERD patients but could no longer be detected in 2 of 6 patients with an initial diagnosis of BE. CONCLUSIONS: With respect to its clinical as well as its endoscopic manifestations, gastroesophageal reflux disease does not appear to be very stable over time. However, in most cases this is due to regression rather than progression of the disease. PMID- 16315126 TI - [Incarcerated Spieghel-Hernie mimicking an acute exacerbation of Crohn's disease]. AB - We report on a 25-year-old woman with long-standing Crohn's disease. Upon admittance to the emergency department, the patient complained of abdominal pain with increasing intensity over the last few days. Clinical examination revealed an abdominal mass in the right lower quadrant, and blood tests showed elevated markers of inflammation. Surprisingly, abdominal ultrasound did not show the suspected complication of Crohn's disease, but rather an incarcerated abdominal wall hernia, which turned out to be a spigelian hernia upon surgical repair. This case stresses the importance of abdominal ultrasound to rule out other diagnoses in patients with chronic inflammatory bowel disease in the emergency setting before starting a potentially dangerous treatment with high-dose steroids. PMID- 16315127 TI - [Pyruvate kinase M2 (tumor M2-PK) as a screening tool for colorectal cancer (CRC). A review of current published data]. AB - INTRODUCTION: Colonoscopy is currently supposed to be the best screening tool for colorectal cancer. However, the acceptance of the population is very poor although it has been included in screening programs in the German health system since 2002. Therefore, evaluation of additional screening tools seems to be of great interest. Recently testing for fecal occult blood (FOBT), genetic alterations or alterations in tumor metabolism (e.g., tumor M2-PK) are under investigation. METHODS: The use of M2-PK measurement in the feces has been reported in 6 studies until today. The data of these studies were analyzed and critically reviewed. RESULTS: The overall sensitivity of M2-PK is 77.9% concerning CRC. Specificity ranges from 74.3-83.3%. Overall sensitivity for adenomas is 45.9%, increasing to 61.1% for adenomas > 1 cm. A high percentage of positive results (90.4%) was also observed in patients with chronic inflammatory bowel disease. CONCLUSIONS: Compared to FOBT or genetic testing the M2-PK test seems to be superior for CRC screening. Concerning handling, effectiveness and analysis, M2-PK seems to be a good possibility for large scale-screening of colorectal carcinoma. It might even be used to detect larger adenomas. Elevated levels of M2-PK in patients with acute and/or chronic inflammatory bowel diseases are probably due to proliferation of epithelial cells and leucocytes in the inflammatory area. PMID- 16315128 TI - [Hepatic hydrothorax]. AB - Hepatic hydrothorax is a rare complication of portal hypertension secondary to liver cirrhosis affecting approximately 5-10% of cirrhotic patients with ascites. Hepatic hydrothorax results from an accumulation of fluid migrating through a diaphragmatic defect from the abdominal cavity into the pleural cavities. The effusion of hepatic hydrothorax is typically transudative whereas the effusion of spontaneous bacterial empyema (SBEM) is exudative. The clinical management of hepatic hydrothorax is equivalent to that of ascites. Patients with persistent hepatic hydrothorax despite fluid and sodium restriction as well as the use of maximally tolerable doses of diuretics and repeated thoracentesis are considered to have refractory hepatic hydrothorax. SBEM is a frequent underlying condition. SBEM occurs in up to 13% of patients with hepatic hydrothorax and should be treated by antibiotic therapy. Refractory hydrothorax is observed in 10% of patients with hepatic hydrothorax. These patients should be considered for transjugular intrahepatic portal systemic shunt (TIPS) placement which is the most effective option for refractory hepatic hydrothorax with response rates ranging up to 80% in most studies. Suitable patients with hepatic hydrothorax should be considered as candidates for liver transplantation. TIPS may help to bridge the time to liver transplantation. PMID- 16315129 TI - Mechanisms of disease: the biophysical interpretation of the ECM affects physiological and pathophysiological cellular behavior. AB - Physiological and pathophysiological migration during the development and systemic spread of tumor cells requires a highly regulated interaction with the extracellular matrix. Sensing of the physical properties of the matrix as well as of forces exerted by the cell or acting on a cell is a prerequisite for productive migration. This review focuses on current concepts of the transmission of a physical stimulus into a biochemical signal in non-neuronal cells. Moreover, we summarize the current concepts on the regulation of affinity-modulation and regulation of protein-turnover for the formation and functionality of adhesion sites with special emphasis on the role of oncogenic signal transduction pathways such as Src family kinases and focal adhesion kinase. PMID- 16315130 TI - [Autoimmune pancreatitis: IgG4 as useful diagnostic approach?]. PMID- 16315132 TI - Clinical aspects of hemochromatosis. AB - The term hemochromatosis is commonly used as synonymous with HFE-associated genetic iron overload but several rarer causes of an identical clinicopathological syndrome have been described in recent years. The most common symptoms are lethargy and arthralgia, and the major complications of end-stage disease are cirrhosis, diabetes, and cardiac and endocrine manifestations. However, with the development of cascade screening for family members of affected probands as well as screening for common diseases at health checks, hemochromatosis is being detected at increasingly early stages, often when there are only biochemical abnormalities. The available evidence from screening studies strongly suggests that approximately 75% of C282Y homozygous subjects have biochemical expression. Hepatic iron overload is present in approximately 56% and 34% of men and women, respectively, advanced hepatic fibrosis in 18.7% and 5.4%, respectively, and cirrhosis in 5.8% and 1.9%, respectively. In subjects with severe expression of the disease, additional modifying genetic mutations have been described including those in hepcidin and hemojuvelin. Treatment is by regular phlebotomy which, if instituted before the development of cirrhosis, results in normal life expectancy. PMID- 16315133 TI - Pathology of hepatic iron overload. AB - The role of liver biopsy in all chronic liver diseases continues to evolve with the emergence of new laboratory and imaging tests. However, the value of histologic examination for fibrosis, parenchymal architectural remodeling, and possible concurrent disease remains relatively unchallenged, including in patients with suspected iron overload. In addition, only histologic evaluation allows detailed analysis of cellular and acinar localization of iron. Routine use of an iron stain for all liver biopsy analysis enables detection of iron when not otherwise suspected. The broad classifications of iron overload include parenchymal ("primary") or reticuloendothelial ("secondary"), and mixed. These classifications, however, serve only as guidelines to differential diagnoses, as will be noted in the following review of pathology of iron overload. Ultimately, the role of the pathologist is to document cellular and acinar localization of iron and to provide a semiquantitative assessment of amount to alert the clinician to the possibility of a significant condition. In this article, histopathologic findings in conditions of iron overload and in several forms of liver disease are discussed. PMID- 16315134 TI - Screening for HFE and iron overload. AB - Type 1 hereditary hemochromatosis is a common disorder of iron overload occurring in individuals homozygous for the C282Y HFE gene mutation. It can be a progressive and fatal condition. Early detection and phlebotomy prior to the onset of cirrhosis can reduce morbidity and normalize life expectancy. It is readily identified through biochemical testing for iron overload using serum transferrin saturation and genetic testing for C282Y homozygosity. General population screening has been waived in preference to targeting high-risk groups such as first-degree relatives of affected individuals and those with clinical features suggestive of iron loading. This screening strategy is likely to continue until uncertainties regarding the natural history of the disease, age related penetrance, and management of asymptomatic individuals are clarified. Potential ethical, legal, and psychosocial issues arising through application of genetic screening programs also must be resolved prior to implementation of general population screening programs. PMID- 16315136 TI - Hepatic iron metabolism. AB - The liver performs three main functions in iron homeostasis. It is the major site of iron storage, it regulates iron traffic into and around the body through its production of the peptide hepcidin, and it is the site of synthesis of major proteins of iron metabolism such as transferrin and ceruloplasmin. Most of the iron that enters the liver is derived from plasma transferrin under normal circumstances, and transferrin receptors 1 and 2 play important roles in this process. In pathological situations, non-transferrin-bound iron, ferritin, and hemoglobin/haptoglobin and heme/hemopexin complexes assume greater importance in iron delivery to the organ. Iron is stored in the liver as ferritin and, with heavy iron loading, as hemosiderin. The liver can divest itself of iron through the plasma membrane iron exporter ferroportin 1, a process that also requires ceruloplasmin. Hepcidin can regulate this iron release through its interaction with ferroportin. PMID- 16315135 TI - Pathophysiology of hereditary hemochromatosis. AB - Hereditary hemochromatosis (HH) encompasses several inherited disorders of iron homeostasis characterized by increased gastrointestinal iron absorption and tissue iron deposition. The most common form of this disorder is HFE-related HH, nearly always caused by homozygosity for the C282Y mutation. A substantial proportion of C282Y homozygotes do not develop clinically significant iron overload, suggesting roles for environmental factors and modifier genes in determining the phenotype. Recent studies have demonstrated that the pathogenesis of nearly all forms of HH involves inappropriately decreased expression of the iron-regulatory hormone hepcidin. Hepcidin serves to decrease the export of iron from reticuloendothelial cells and absorptive enterocytes. Thus, HH patients demonstrate increased iron release from these cell types, elevated circulating iron, and iron deposition in vulnerable tissues. The mechanism by which HFE influences hepcidin expression is an area of current investigation and may offer insights into the phenotypic variability observed in persons with mutations in HFE. PMID- 16315137 TI - Hepatotoxicity of iron overload: mechanisms of iron-induced hepatic fibrogenesis. AB - While iron is a vital requirement for normal cellular physiology, excessive intestinal absorption of iron as seen in hemochromatosis leads to its deposition in parenchymal cells of various organs such as the liver, heart, and pancreas, resulting in cellular toxicity, tissue injury, and organ fibrosis. Cellular injury is induced by iron-generated oxyradicals and peroxidation of lipid membranes. In the liver, lipid peroxidation results in damage to hepatocellular organelles, such as mitochondria and lysosomes, which is thought to contribute to hepatocyte necrosis and apoptosis, and ultimately lead to the development of hepatic fibrogenesis. Hepatic stellate cells are central to the development of hepatic fibrosis, as they can be activated into collagen-producing myofibroblasts. Numerous potential stimuli associated with hepatic iron overload and iron-induced hepatocellular injury have been assessed in an attempt to explain stellate cell transformation in hemochromatosis. Stellate cell activation and fibrosis appear to be regulated by a series of events involving cellular interaction between resident and nonresident cells of the liver, the sequestration of free iron versus the transport and storage of mobilizable iron, and extracellular matrix remodeling as well as intracellular signaling events associated with inflammatory and fibrogenic cytokines. PMID- 16315138 TI - Non-HFE hemochromatosis. AB - The term "non-HFE hemochromatosis" (non-HFE HC) refers to several phenotypically similar but genetically distinct forms of hereditary hemochromatosis affecting individuals without pathogenic mutations of HFE. The involved genes are, sinsu strictu, transferrin receptor 2 (TfR2), hemojuvelin (HJV), and hepcidin (HAMP). Non-HFE HC share common pathogenic and clinical features with HFE HC. However, depending on the role of the affected gene in iron trafficking, the clinical onset may be earlier and phenotypic expressivity more severe than classic HC. Other forms of hereditary iron overload have distinct pathogenesis and phenotype. The most prevalent of these forms is "ferroportin disease," characterized by autosomal dominant trait, predominant reticuloendothelial cell iron overload, and mild organ damage. Non-HFE HC gene products, while responsible for rarer cases of HC as compared with HFE, are much more central than HFE in human iron homeostasis and understanding their function will greatly advance our comprehension of iron trafficking in health and disease. PMID- 16315139 TI - Iron in nonhemochromatotic liver disorders. AB - Iron is essential for cellular functions, but in excessive amounts it is toxic to cells. The harmful effects are related to increased oxidative stress and production of reactive oxygen species causing oxidative damage to lipids, proteins, and nucleic acids. Heavy iron overload as occurs in primary and secondary hemochromatosis can cause fibrosis of various parenchymal organs such as the liver, heart, and pancreas. Lesser degrees of hepatic iron deposition are also associated with, and seem to be risk factors for, certain nonhemochromatotic liver diseases. Porphyria cutanea tarda is associated with hepatic iron overload and responds to iron-reduction therapy. Other recent evidence indicates that the prevalence of HFE gene mutations is increased in chronic viral hepatitis and that patients with chronic hepatitis C harboring especially the C282Y mutation are more likely to suffer from advanced hepatic fibrosis or cirrhosis and to do so at younger ages. In this article we review selected nonhemochromatotic disorders in which iron can play an important comorbid role. PMID- 16315140 TI - Rapid analysis of controlled substances using desorption electrospray ionization mass spectrometry. AB - The recently developed technique of desorption electrospray ionization (DESI) has been applied to the rapid analysis of controlled substances. Experiments have been performed using a commercial ThermoFinnigan LCQ Advantage MAX ion-trap mass spectrometer with limited modifications. Results from the ambient sampling of licit and illicit tablets demonstrate the ability of the DESI technique to detect the main active ingredient(s) or controlled substance(s), even in the presence of other higher-concentration components. Full-scan mass spectrometry data provide preliminary identification by molecular weight determination, while rapid analysis using the tandem mass spectrometry (MS/MS) mode provides fragmentation data which, when compared to the laboratory-generated ESI-MS/MS spectral library, provide structural information and final identification of the active ingredient(s). The consecutive analysis of tablets containing different active components indicates there is no cross-contamination or interference from tablet to tablet, demonstrating the reliability of the DESI technique for rapid sampling (one tablet/min or better). Active ingredients have been detected for tablets in which the active component represents less than 1% of the total tablet weight, demonstrating the sensitivity of the technique. The real-time sampling of cannabis plant material is also presented. PMID- 16315141 TI - NMR conformational analysis of proadrenomedullin N-terminal 20 peptide, a proangiogenic factor involved in tumor growth. AB - The preferred conformation of Proadrenomedullin N-Terminal 20 Peptide (PAMP; ARLDVASEFRKKWNKWALSR-amide) has been determined using 1H and 13C two-dimensional nuclear magnetic resonance (NMR) spectroscopy and molecular modeling. PAMP is a peptide that has various physiological functions, including its role as a proangiogenic factor in facilitating tumor growth and its inhibitory effect on catecholamine secretion at nicotinic receptors. The preferred conformation of PAMP was determined in a helix-inducing trifluoroethanol and water (TFE/H2O) solution, and in a membrane-mimetic sodium dodecylsulfate-d25 (SDS) micellar solution. The secondary structure consists of an alpha-helix for residues Arg2 to Arg20 in TFE/H2O solution and an alpha-helix for residues Arg2 to Ala17 in SDS solution. We postulate that the polar charged residues Arg2, Lys12, and Arg20 are responsible for the initial interaction of the peptide with the micelle, and that this is followed by the binding of the hydrophobic residues Leu3, Val5, Phe9, Trp13, and Trp16 to the micellar core. The three C-terminal amino acid residues adopt an extended structure in SDS, suggesting that they are important in receptor recognition and binding. This is supported by truncation studies done by Mahata et al. (Hypertension, 1998, Vol. 32, pp. 907-916), which show the importance of the C-terminal in physiological activity. Furthermore, Belloni et al. (Hypertension, 1999, Vol. 33, pp. 1185-1189), and Martinez et al. (Cancer Research, 2004, Vol. 64, pp. 6489-6494) suggested that the N-terminal was also important in PAMP activity. However, no differences in conformational preference of the N-terminal were observed between the two solvent systems. PMID- 16315142 TI - Characterization of the acidic and basic limbs of a bell-shaped pH profile in the inhibitory activity of bromelain inhibitor VI. AB - Bromelain inhibitor VI (BI-VI) is a cysteine proteinase inhibitor from pineapple stem and a unique two-chain inhibitor composed of two distinct domains. BI-VI's inhibitory activity toward the target enzyme bromelain is maximal at pH 4 and shows a bell-shaped pH profile with pKa values of about 2.5 and 5.3. This pH profile is quite different from that of bromelain, which is optimally active around pH 7. In the present article, to characterize the acidic limb, we first expressed the recombinant inhibitors designed to lose two putative hydrogen bonds of Ser7(NH)-Asp28(beta-CO2H) and Lys38(NH)-Asp51(beta-CO2H) and confirmed the existence of the hydrogen bonds by two-dimensional nuclear magnetic resonance (NMR). Moreover, it was revealed that these hydrogen bonds are not the essential electrostatic factor and some ionizable groups would be responsible for the acidic limb in the pH-inhibition profile. On the other hand, to characterize the basic limb, we examined the pH-dependent inhibition using the cysteine proteinase papain, some of whose properties differ from those of bromelain, and compared the data with the corresponding data for bromelain. The result suggests that the basic limb would be affected by some electrostatic factors, probably some carboxyl groups in the target proteinase. PMID- 16315143 TI - Determination of intrinsic hydrophilicity/hydrophobicity of amino acid side chains in peptides in the absence of nearest-neighbor or conformational effects. AB - Understanding the hydrophilicity/hydrophobicity of amino acid side chains in peptides/proteins is one the most important aspects of biology. Though many hydrophilicity/hydrophobicity scales have been generated, an "intrinsic" scale has yet to be achieved. "Intrinsic" implies the maximum possible hydrophilicity/hydrophobicity of side chains in the absence of nearest-neighbor or conformational effects that would decrease the full expression of the side chain hydrophilicity/hydrophobicity when the side chain is in a polypeptide chain. Such a scale is the fundamental starting point for determining the parameters that affect side-chain hydrophobicity and for quantifying such effects in peptides and proteins. A 10-residue peptide sequence, Ac-X-G-A-K-G-A-G-V-G-L amide, was designed to enable the determination of the intrinsic values, where position X was substituted by all 20 naturally occurring amino acids and norvaline, norleucine, and ornithine. The coefficients were determined by reversed-phase high-performance liquid chromatography using six different mobile phase conditions involving different pH values (2, 5, and 7), ion-pairing reagents, and the presence and absence of different salts. The results show that the intrinsic hydrophilicity/hydrophobicity of amino acid side chains in peptides (proteins) is independent of pH, buffer conditions, or whether C(8) or C(18) reversed-phase columns were used for 17 side chains (Gly, Ala, Cys, Pro, Val, nVal, Leu, nLeu, Ile, Met, Tyr, Phe, Trp, Ser, Thr, Asn, and Gln) and dependent on pH and buffer conditions, including the type of salt or ion-pairing reagent for potentially charged side chains (Orn, Lys, His, Arg, Asp, and Glu). PMID- 16315145 TI - Diogene syndrome, a clinical presentation of fronto-temporal dementia or not? PMID- 16315146 TI - Residual symptoms in older patients treated for major depression. AB - OBJECTIVE: The purpose of this study was to identify residual symptoms in a sample of older adults treated for major depression and compare individual symptoms present at baseline with those at three months by remission status. METHODS: The sample was comprised of 229 patients with DSM-IV major depression who were participants in the NIMH Mental Health Clinical Research Center at Duke University. Symptoms were measured using the Montgomery-Asberg Depression Rating Scale (MADRS). RESULTS: At three months, 86 patients (37.6%) had remitted, or had a MADRS score less than or equal to 9. In the remitted group, the most frequently reported symptoms at three months were inner tension and lassitude. Among nonremitters, the most frequently reported symptoms were reported and apparent sadness, as well as lassitude and inner tension. In the sample as a whole, the symptoms most likely to be present at baseline but not three months were pessimistic and suicidal thoughts, while the most frequently reported emergent symptoms were reduced appetite and inner tension. Patients were much more likely to no longer have a particular symptom than to acquire a new symptom. Overall, the symptoms present at three months were not severe in either group. CONCLUSIONS: In older adults treated for major depression, residual symptoms at three months may include emergent symptoms as well as persistent symptoms, and are likely to include symptoms of anxiety as well as sadness. These findings have clinical implications for the treatment of late-life depression. PMID- 16315147 TI - Behavioural and psychological signs and symptoms of dementia across cultures: current status and the future. AB - BACKGROUND: Behavioural and psychological symptoms of dementia (BPSD) have been poorly studied in developing countries, in ethnic minority groups in a given country and in cross-national studies. METHODS: The literature on BPSD from developing countries, ethnic minority groups in a given country and cross national studies was examined. RESULTS: There is emerging literature on BPSD from the settings was studied. These studies provide useful preliminary data on the prevalence and correlates of BPSD. Moreover, the data illustrate possible cross cultural differences in BPSD and their correlates. A number of instruments measuring either individual BPSD within a BPSD domain, measuring features of a BPSD domain or features of a range of BPSD domains have been developed in languages other than English for use in developing countries and ethnic minority groups in a given country. CONCLUSIONS: There is a need for methodologically similar and uniform studies of BPSD across countries and ethnic groups in a given country using appropriately validated instruments. It is suggested that a consensus should be reached by researchers on the best instrument(s) to be developed in languages other than English for use in these settings and, in turn, these instruments should be developed using appropriate methodology. This could allow identification of the genetic and environmental aetiology of BPSD and the influence of gene-environment interaction. PMID- 16315148 TI - The management of inappropriate vocalisation in dementia: a hierarchical approach. AB - BACKGROUND: Patients with dementia can demonstrate noisy behaviours such as screaming, repetitive speech, moaning and singing. Such behaviours can be grouped under the title of "Inappropriate Vocalisation" which is used in this article to describe any noise making which impacts detrimentally upon patients, families or those in a caring role. Inappropriate vocalisation is notoriously difficult to treat and clinicians may have to rely on a "trial and error" approach when attempting to limit the distress it causes. OBJECTIVES: This paper applies a hierarchical approach to Behavioural and Psychological Symptoms of Dementia (BPSD) in attempting to tackle inappropriate vocalisation systematically. Nine steps are ranked within three stages, with evidence for each intervention being considered sequentially in terms of the likelihood for success as the authors attempt to review the relevant literature. METHODS: The authors performed a search of the medline database using the key words "screaming," "shouting," "persistent vocalisation" and "inappropriate vocalisation" combined with the key word "dementia". Further searches of the references of medline generated articles revealed a second group of articles of use in this review. RESULTS: The search generated 53 reviews, research papers, case studies or letters, 36 of which are referenced in this article and the remaining references drawn from reading by the authors around the subject and related problems. CONCLUSION: Interventions for inappropriate vocalisation in dementia are not limited to medication. Environmental factors and behavioural approaches are considered ahead of pharmacotherapy as the concepts of Underlying Illness, Hidden Meaning and Empirical Treatments are used to structure consideration of important, and sometimes overlooked, issues including pain, depression and overall level of stimulation. PMID- 16315149 TI - The TE4D-Cog: a new test for detecting early dementia in English-speaking populations. AB - BACKGROUND: The screening test usually used to detect dementia (Mini Mental State Examination, MMSE) is limited by a ceiling effect and high false positive rates, as are other similar instruments. There is therefore a need for a more sensitive and specific screening tool to aid early detection and diagnosis of dementia. OBJECTIVE: The hypothesis of the study was that the TE4D-Cog would be more sensitive and specific than the MMSE in detecting mild cognitive impairment in patients with AD. METHOD: The TE4D (Test for the Early Detection of Dementia from Depression) was adapted from its original German version for English-speaking populations. This new scale (the TE4D-Cog) was then administered together with the MMSE and the cognitive subscale of the Alzheimer's disease Assessment Scale (ADAS-Cog) to 178 people with a diagnosis of Alzheimer's disease and 25 cognitively intact comparators. The sensitivity and specificity in detecting dementia of the TE4D-Cog and the MMSE were compared in those with mild dementia and those without dementia. RESULTS: The TE4D-cog had high sensitivity with an acceptable specificity and low false positive rate. It also had good concurrent validity, high inter-rater reliability, good internal consistency and strong predictive validity. CONCLUSIONS: The TE4D-Cog is easy to administer, short and acceptable. Results are independent of age, gender and level of education. The TE4D-Cog may therefore be a useful alternative to the MMSE as a dementia screening instrument. PMID- 16315150 TI - Discontinuation of rivastigmine in routine clinical practice. AB - BACKGROUND: Rivastigmine is used for symptomatic treatment of mild-to-moderately severe Alzheimer's Dementia (AD). We investigated the frequency of and reasons for rivastigmine discontinuation in clinical practice and possible predictive factors for discontinuation within the first six months after starting therapy. METHODS: A retrospective cohort study was performed in rivastigmine users, who started therapy in a naturalistic setting. A nurse supported a part of the studied cohort, as this was introduced during the study period. Reasons for discontinuation were investigated, including therapy discontinuation if the Maximum Achieved Dose (MAD) was below 6 mg daily. Predictors of discontinuation within the first half year were investigated by logistic regression analysis. RESULTS: Baseline Mini-Mental-State-Examination (MMSE) of included patients (n = 154) was 20.1, mean age was 78.4 years and 70% was female. Within 6 months, 61 users (39.6%) discontinued therapy, primarily (59.0%) for adverse events. Thereafter, the main reason for discontinuation was non-response according to clinimetrics. A MAD during the titration phase of 1.5-4.5 mg/day and absence of nurse support are significantly related to discontinuation within 6 months. CONCLUSIONS: Rivastigmine is primarily discontinued within the first six months for intolerable adverse events and thereafter mainly for ongoing deterioration. A MAD of 1.5-4.5 mg/day and the absence of nurse support are independently related to discontinuation of rivastigmine within the initial 6 months. PMID- 16315151 TI - Delirium-O-Meter: a nurses' rating scale for monitoring delirium severity in geriatric patients. AB - BACKGROUND: Delirium is a common psychiatric disorder in general hospital elderly patients. Several delirium screening tests exist. Few nurse based delirium severity measures are available. The aim of this study was to evaluate the Delirium-O-Meter, a new nurses' behavioural rating scale that is an efficient and sensitive measure of delirium severity. METHODS: Analysis of cross sectional and repeated assessments data. Participants were 92 elderly general hospital patients; 56 with delirium, 24 with dementia or other cognitive disturbances (no delirium) and 12 with other psychiatric disorders or no mental disorder. Measures were the Delirium-O-Meter (DOM), Delirium Rating Scale-Revised version (DRS-R 98), Delirium Observation Scale (DOS), Behavioural observation scale for geriatric inpatients (GIP) and Mini Mental State Examination (MMSE). RESULTS: The majority of DOM items show a (near-) normal score distribution. Reliability of the DOM was high; Cronbach's alpha values ranged from 0.87-0.92; Intra Class Correlation (ICC) range was 0.84-0.91 for total scores and 0.40-0.97 for item scores. Factor analysis produced a 'Cognitive/Motivational' factor explaining almost half of variance and a smaller 'Psychotic/Behavioural' factor. The two factor model results support the conceptual distinction between hyperactive and hypoactive delirium. DOM observations differentiated delirium from non delirium patients. DOM total scores were highly related to the DRS-R-98, DOS, MMSE and GIP apathy and cognitive sub scales, but less so to the GIP affective disturbances subscale, indicating convergent and divergent validity. Temporal difference scores calculated for DRS-R-98 and DOM assessments on subsequent days were also highly related (rho = 0.80-0.95). CONCLUSIONS: The newly constructed DOM is a brief and valid nurses' behavioural rating scale that can be useful for measuring different aspects of delirium and for efficiently monitoring delirium severity in elderly patients. PMID- 16315152 TI - Anatomy, surgery, and thinking. PMID- 16315153 TI - Mental health of older world leaders. PMID- 16315157 TI - Comparative performance of long and short forms of the Geriatric Depression Scale in mildly demented Chinese. AB - OBJECTIVE: This study aims to investigate if a brief version (four items) of the Geriatric Depression Scale works equally well with mildly demented as with nondemented patients in young-old and old-old persons, and to compare its diagnostic performances with those of the 15- and the 30-item version of the scale. METHOD: Four hundred and forty-two older persons were given a GDS interview and received an independent psychiatric evaluation. Receiver operating characteristic curves were separately plotted for the young-old demented, old-old demented, young-old nondemented, and old-old nondemented. RESULTS: The four-item version yielded comparable performance to the 15- and the 30-item version of the GDS, regardless of age and dementia status. It is reasonably robust to the effects of age and mild dementia, whether alone or in combination. Nonetheless, among the old-old demented, only a third of those tested positive were actually positive, but this problem was not specific to the four-item version. CONCLUSIONS: The four-item version can be used in lieu of the longer versions to conserve clinical and research resources where appropriate, and a cutoff of 1/2 can be applied across the board. Little, if any, information is lost when this brief version is used instead of the longer ones. PMID- 16315158 TI - Why and how antipsychotic drugs are used in 40 Sydney nursing homes. AB - BACKGROUND: For decades there has been a high use of antipsychotic medication in nursing homes, though recently the use of typical antipsychotics has reduced while use of atypical antipsychotic medication has increased. The Australian government subsidises use of the latter in management of schizophrenia, and since April 2005 has subsidised use of risperidone (but still not the other atypicals) in cases of dementia with behavioural disturbance. This study was designed to examine the pattern of, and reasons for, antipsychotic use in a range of Sydney nursing homes. METHODS: Clinical files and medication cards in 40 Sydney nursing homes were examined in late 2003 by a research nurse, who recorded documented diagnoses, blood glucose levels and use of medication in the previous 4 weeks. RESULTS: Antipsychotics were prescribed for 577 (25.1%) of the 2302 residents. Of these, 114 had schizophrenia. Of the 2302, 6.1% and 15.6%, respectively, had documented diagnoses of schizophrenia and/or diabetes. Of those with schizophrenia, 13.6% had diabetes. Two-thirds of those given antipsychotics had dementia or cerebral disease and not schizophrenia. Two-thirds of the antipsychotic prescriptions for residents with schizophrenia, and also for those without, were for atypicals (n = 200 olanzapine, 174 risperidone, 19 quetiapine). Blood glucose measurements had been recorded in less than 45%. CONCLUSIONS: In Sydney in 2003, most (80%) of the nursing home residents for whom antipsychotics were prescribed did not have a diagnosis of schizophrenia. The records suggested that insufficient attention had been given to the possibility that use of certain antipsychotics can be associated with impaired glucose metabolism. PMID- 16315159 TI - Risperidone for psychosis of Alzheimer's disease and mixed dementia: results of a double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate the efficacy and safety of low-dose risperidone in treating psychosis of Alzheimer's disease (AD) and mixed dementia (MD) in a subset of nursing-home residents who had dementia and aggression and who were participating in a randomized placebo-controlled trial of risperidone for aggression. METHOD: This post-hoc analysis included only patients diagnosed with AD or MD with psychosis, defined by a score of >or= 2 on any item of the Behavioral Pathology of Alzheimer's Disease (BEHAVE-AD) psychosis subscale at both screening and baseline. Co-primary efficacy endpoints were changes in scores on BEHAVE-AD psychosis subscale and Clinical Global Impression of Change (CGI-C). RESULTS: Overall, 93 patients (46 risperidone and 47 placebo) fulfilled the psychosis of AD criteria. Mean change at endpoint in BEHAVE-AD psychosis subscale with risperidone was superior to placebo (-5.2 vs -3.3; p = 0.039). Distribution of CGI-C at endpoint also favoured risperidone (p < 0.001). The superior improvement with risperidone compared with placebo occurred as early as the first two weeks and persisted to the end of the treatment period. At endpoint, 59% of risperidone-treated patients were responders (i.e. were 'very much' or 'much' improved) compared with 26% of patients receiving placebo. The mean risperidone dose was 1.03 +/- 0.61 mg/day. Twelve weeks of treatment were completed by 37 patients treated with risperidone (80%) and 35 with placebo (74%). A total of 46 (98%) placebo- and 44 (96%) risperidone-treated patients experienced at least one adverse event, with only somnolence occurring more frequently in the risperidone group. CONCLUSION: Risperidone effectively reduces psychosis and improves global functioning in elderly patients with moderate-to-severe psychosis of AD and MD. PMID- 16315160 TI - Review of assessment and treatment of PTSD among elderly American armed forces veterans. AB - BACKGROUND: The number of elderly combat veterans is steadily increasing in the US and estimates project that a notable percentage of these veterans experience symptoms of posttraumatic stress disorder (PTSD). Limited data exist specifically related to prevalence, assessment, and treatment of PTSD among the elderly veteran population. OBJECTIVE: This review summarizes the available research related to difficulties in assessment with the elderly American Armed Forces veteran population. In addition, both psychotherapeutic and pharmacological treatment interventions for PTSD are discussed. METHODS: A literature search was conducted using PsycINFO, Medline, and the National Center for PTSD's PILOTS database. RESULTS: Evidence suggests that elderly veterans generally present more somatic symptoms of PTSD. Medical and psychological comorbidities, such as depression, substance abuse, or cognitive deficits can further complicate the assessment process. Cut-scores for existing instruments need to be further established with elderly veterans. Use of exposure therapies with the elderly has not been adequately researched and mixed results have been obtained for supportive therapy for treatment of PTSD. Controlled research investigating pharmacological interventions for PTSD with the elderly is also limited. CONCLUSION: Evidence suggests that some psychotherapeutic and pharmacological interventions already utilized with younger individuals may be useful with the elderly veteran population. However, research indicates that modifications may be required for working with the elderly population and further research in the areas of assessment and treatment are necessary. PMID- 16315161 TI - Assessing early to late stage dementia: the TSI and BANS-S scales in the nursing home. AB - BACKGROUND: The traditional assessment tools for dementia, such as the MMSE, have only limited ability to follow subjects with severe dementia because they show a floor effect. Specific observational and performance-based instruments were recently developed. OBJECTIVES: To directly compare an observational scale to a performance-based instrument in moderate to severe dementia. METHODS: We compared a slightly modified version of the performance-based Test for Severe Impairment (mTSI) to the observer-based Bedford Alzheimer Nursing Severity Scale (BANS-S). Both scales were administered, together with the Mini-Mental State Examination (MMSE) and the Clinical Dementia Rating scale (CDR), to a nursing-home sample of 130 women suffering from different types of dementia (CDR range:1-4; MMSE range:0 18), defined according to DSM-IV criteria. Mean age was 86.9 +/- 7.3 years and mean education was 2.7 +/- 1.1 years. RESULTS: The BANS-S could be applied to all patients, the mTSI to 87 subjects (66.9%). Mean mTSI score decreased progressively from CDR stage 2 to CDR stage 4, whereas no difference was detectable between CDR stages 1 and 2. By contrast, the BANS-S was not significantly different for CDR stages 1 to 3, and the mean BANS-S score worsened only in CDR stage 4. Results were similar for AD and non AD dementia. Both scales were independent from age and education and their test-retest and inter-rater reliabilities were satisfactory. CONCLUSION: The mTSI looks promising in the moderate-to-severe range, whereas the BANS-S seems more useful in the very late stage of dementia. However, neither scale was optimal and additional instruments should be tested in future studies. PMID- 16315162 TI - Molecular mechanism of the solid-state fluorescence behavior of the organic pigment yellow 101 and its derivatives. PMID- 16315163 TI - Direct conversion of linoleic acid over silver catalysts in the presence of H2: an unusual way towards conjugated linoleic acids. PMID- 16315165 TI - Recent advances in the application of capillary electrophoresis with mass spectrometric detection. AB - This review gives an overview of applications of CE coupled to MS detection published in the literature of the last three years. The works discussed in this paper comprise a wide range of different fields of application. These include important sections such as the analysis of biomolecules, the analysis of pharmaceuticals and their metabolites in different matrices, environmental analysis, and also investigations on the composition of technical products. PMID- 16315166 TI - Nanoparticle-based pseudostationary phases in capillary electrochromatography. AB - During the past decades, research has been performed to enhance selectivity in CE by introducing different types of additives into the electrolyte. Research concerning this has taken many directions, especially during the last 5 years. A promising technique, which benefits from no packing or frits, is to use nanoparticles as the pseudostationary phase (PSP) in CEC. PSPs have the advantage of introducing a novel interaction phase for every analysis, which greatly simplify column exchange and circumvent contamination inherited from complex mixtures, e.g., biological samples. The field of nanoparticle-based PSPs used in CEC is covered in this review. The term CEC will be used consequently throughout this review, although some authors used the term EKC to categorize their work. Important requirements for the nanoparticles used and possible reasons for band broadening will be discussed. Applications with silica nanoparticles, polymer nanoparticles, molecularly imprinted polymer nanoparticles, gold nanoparticles, dendrimers, and polymeric surfactants as PSP will also be discussed. PMID- 16315167 TI - Recent advances in capillary electrophoresis and capillary electrochromatography of pollutants. AB - Recent advances in the CE and CEC separation, detection, and sample preparation methodologies applied to the determination of a variety of compounds having current or potential environmental relevance have been overviewed. The reviewed literature has illustrated the wide range of CE applications, indicating the continuing interest in CE and CEC in the environmental field. PMID- 16315168 TI - Recent applications in capillary electrochromatography. AB - In this review research papers on the application of CEC are summarized that have been published between May 2003 and May 2005. First, a short overview is given of trends and developments in CEC that may increase the applicability of the separation technique. Next, application-oriented research using CEC is described in biochemical studies, including proteomics and genomics, in the analysis of food and natural products, and in pharmaceutical, industrial, and environmental analysis. PMID- 16315169 TI - Pharmacokinetic applications of capillary electrophoresis: a review on recent progress. AB - This article covers recent publications from 2003 to 2005 on the subject of pharmacokinetic applications of CE. Many analytical methods were validated and more importantly, they were shown to have sufficient sensitivities to access pharmacokinetic data on different models. Because of unique advantages, such as simplified sample preparation methods, small sample amount required, high separation power, and speedy analysis, CE-based assays were found to gain popularity not only as a second method but also as a major method for many pharmacokinetic studies. PMID- 16315171 TI - Application of horizontal staircase electrophoresis in agarose minigels to the random intergenic spacer analysis of clinical samples. AB - The random intergenic spacer analysis is a recently developed technique for the study of microbial populations. The bacterial intergenic spacer (ITS) is located between 16S rRNA and 23S rRNA genes and presents different length and sequence among bacterial species. Therefore, the amplicons can be separated by electrophoresis commonly performed at low voltage during several hours. Although this technique is especially useful for unculturable microorganisms, it has not been applied before to clinical sample analysis. As these samples have a limited number of bacterial species, the size of the gels may be reduced to facilitate their handling and to reduce the running time. To obtain maximum separation among the ITS bands, we analysed in this work different electrophoretical conditions including staircase electrophoresis, a technique based on the application of several voltage steps. The results obtained showed a different behaviour of the electrical resistance during the performance of submarine horizontal and vertical staircase electrophoresis. In the first case the resistance decreased during most of the running time whereas in the second case it increased. Here, we show that the performance of horizontal staircase electrophoresis reduces the running time more than 80% with respect to conventional electrophoresis at low voltages. This procedure was applied to the separation of ITS bands from bacterial DNA present in a tissue from a vocal cord biopsy. The sequencing of these bands allowed their identification. This new procedure may be very useful in the rapid diagnosis of bacteria present in human, animal and plant tissues. PMID- 16315172 TI - Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen as fluorescence inducing agent. AB - The fast separation of oligonucleotide (oligos) sizing marker by CE using OliGreen and including effects due to the concentration of separation medium and urea denaturant is presented. OliGreen dye is found to be more sensitive than ethidium bromide (by a factor of about 6 based on S/N considerations) for the oligos' separations. Higher concentration of F127 in 1xTris-boricacid-EDTA (TBE) up to 30% w/v leads to better resolution of oligos separations. The addition of urea into the separation medium decreases the sensitivity. With an optimized running condition, the oligos sizing marker could be successfully separated with 1-base resolution within 1.3 min by using 30% w/v F127/1xTBE solution as the separation medium at an applied electric field of 800 V/cm in a 3 cm long capillary, the fastest capillary gel electrophoresis separation with high resolution reported to date for oligos in the similar size range. PMID- 16315173 TI - Study on the enhancement of Ru(bpy)3(2+) electrochemiluminescence by nanogold and its application for pentoxyverine detection. AB - In this work, CE separation with end-column Ru(bpy)3(2+) ECL detection for the quantitative determination of pentoxyverine was firstly performed. The experimental conditions, such as the applied potential, injection voltage, injection time, and the pH of separation buffer, were discussed in detail. Gold nanoparticles were found to enhance the ECL intensity at an appropriate volume ratio of nanogold with Ru(bpy)3(2+) but without changing their nanoproperties proved by transmission electron microscopy (TEM) and UV-vis spectra. The detection limits with or without nanogold were 6 nM and 0.6 microM, respectively. Successful separation of pentoxyverine, chlorpheniramine, and lidocaine was achieved. This method was also applied to monitor drug binding with HSA, and the binding constant for pentoxyverine was estimated as 1.8 x 10(3)/M. PMID- 16315174 TI - Biomarker discovery in rat plasma for estrogen receptor-alpha action. AB - To support in vivo screening efforts for estrogen receptor (ER) subtype selective therapeutic agents, we initiated work to discover surrogate markers (biomarkers) in blood plasma that would change in response to ER subtype-specific action. We used a proteomic approach employing strong anion exchange chromatography (SAX), PAGE, and MS to identify potential plasma markers for selective ER-alpha action. The methodology was used to compare blood from vehicle-treated rats to blood from rats treated with either 17beta-estradiol (an ER-alpha/ER-beta agonist) or compound 1 (17alpha-ethynyl-[3,2-c]pyrazolo-19-nor-4-androstene-17beta-ol, an ER alpha-selective agonist). Blood samples were first fractionated by SAX to separate fractions containing dominant common plasma proteins from fractions enriched for less-abundant plasma proteins. 1-D PAGE analysis of fractions depleted of dominant plasma proteins revealed treatment-specific changes in protein profiles. Protein bands that changed reproducibly in response to ER-alpha action were excised from the gel, separated by capillary LC, and identified by microspray ESI-MS. Using this method, the plasma levels of two proteins, transthyretin and apolipoprotein E, were shown to decrease in response to ER alpha agonism. The method lacked the sensitivity to identify the known, 1000-fold less-abundant, estrogenic marker prolactin (PRL). However, using a commercial RIA and immunoblots, we showed that PRL levels increase significantly in response to treatment with the ER-alpha selective agonist, compound 1. PMID- 16315175 TI - Large-scale muLC-MS/MS for silver- and Coomassie blue-stained polyacrylamide gels. AB - 2-DE combined with LC-MS/MS has become a routine, reliable protein separation and identification technology for proteome analysis. The demand for large-scale protein identifications after 2-DE separation requires a sensitive and high throughput LC-MS/MS method. In this report, a simple, splitless, fully automated capillary LC-MS/MS system was described for the large-scale identification of proteins from gels stained with either silver or CBB. The gel samples were digested and peptides were extracted using an in-gel digestion workstation. The peptides were automatically introduced into a capillary column by an autosampler connected to an HPLC pump. A nanoLC pump was then used to deliver the gradient and elute the peptides from the capillary column directly into an LCQ IT mass spectrometer. Neither a peptide trapping setting nor a flow split is needed in this simple setup. The collected MS/MS spectra were then automatically searched by SEQUEST, and filtered and organized by DTASelect. Hundreds of silver-stained or CBB-stained Shewanella oneidensis, Geobacter sulfurreducens, and Geobacter metallireducens proteins separated by denaturing or nondenaturing 2-DE were digested and routinely analyzed using this fully automated muLC-MS/MS system. High peptide hits and sequence coverage were achieved for most CBB-stained gel spots. About 75% of the spots were found to contain multiple proteins. Although silver staining is not commonly thought to be optimal for MS analysis, protein identifications were successfully obtained from silver-stained 2-DE spots detected using methods with and without formaldehyde for protein fixation. PMID- 16315176 TI - Software-induced variance in two-dimensional gel electrophoresis image analysis. AB - Experimental variability in 2-DE is well documented, but little attention has been paid to variability arising from postexperimental quantitative analyses using various 2-DE software packages. The performance of two 2-DE analysis software programs, Phoretix 2D Expression v2004 (Expression) and PDQuest 7.2 (PDQuest), was evaluated in this study. All available background subtraction and smoothing algorithms were tested using both data generated from one single 2-DE gel image, thus excluding experimental variance, and with authentic sets of replicate gels (n = 5). A slight shift of the image boundaries (the "cropping area") caused both programs to induce variance in protein spot quantification of otherwise identical gel images. The resulting variance for PDQuest (CV(mean) = 8%) was approximately twice that for Expression (CV(mean) = 4%). In authentic sets of replicate 2-DE gels (n = 5), the experimental variance confounded the software-induced variance to some extent. However, Expression still outperformed PDQuest, which exhibited software-induced variance as high as 25% of the total observed variance. Surprisingly, the complete omission of background subtraction algorithms resulted in the least amount of software-based variance. These data indicate that 2-DE gel analysis software constitutes a significant source of the variance observed in quantitative proteomics, and that the use of background subtraction algorithms can further increase the variance. PMID- 16315177 TI - A hydroponic rice seedling culture model system for investigating proteome of salt stress in rice leaf. AB - By using an in vivo hydroponic rice seedling culture system, we investigated the physiological and biochemical responses of a model rice japonica cultivar Nipponbare to salt stress using proteomics and classical biochemical methods. Yoshida's nutrient solution (YS) was used to grow rice seedlings. YS-grown 18-day old seedlings manifested highly stable and reproducible symptoms, prominently the wilting and browning of the 3rd leaf, reduced photosynthetic activity, inhibition in overall seedling growth, and failure to develop new (5th) leaf, when subjected to salt stress by transferring them to YS containing 130 mM NaCl for 4 days. As leaf response to salt stress is least investigated in rice by proteomics, we used the 3rd leaf as source material. A comparison of 2-DE protein profiles between the untreated control and salt-stressed 3rd leaves revealed 55 differentially expressed CBB-stained spots, where 47 spots were increased over the control. Of these changed spots, the identity of 33 protein spots (27 increased and 5 decreased) was determined by nESI-LC-MS/MS. Most of these identified proteins belonged to major metabolic processes like photosynthetic carbon dioxide assimilation and photorespiration, suggesting a good correlation between salt stress-responsive proteins and leaf morphology. Moreover, 2-DE immunoblot and enzymatic activity analyses of 3rd leaves revealed remarkable changes in the key marker enzymes associated with oxidative damage to salt stress: ascorbate peroxidase and lipid peroxidation were induced, and catalase was suppressed. These results demonstrate that hydroponic culture system is best suited for proteomics of salt stress in rice seedling. PMID- 16315178 TI - Two-dimensional gel electrophoresis maps of the proteome and phosphoproteome of primitively cultured rat mesangial cells. AB - Mesangial cells (MC) play an important role in maintaining the structure and function of the glomerulus. The proliferation of MC is a prominent feature of many kinds of glomerular disease. The first reference 2-DE maps of rat mesangial cells (RMC), stained with silver staining or Pro-Q Diamond dye, have been established here to describe the proteome and phosphoproteome of RMC, respectively. A total of 157 selected protein spots, corresponding to 118 unique proteins, have been identified by MALDI-TOF-MS or LC-ESI-IT-MS/MS, in which 37 protein spots representing 28 unique proteins have also been stained with Pro-Q Diamond, indicating that they are in phosphorylated forms. All the identified proteins were bioinformatically annotated in detail according to their physiochemical characteristics, subcellular location, and function. Most of the separated or identified protein spots are distributed in the area of mass 10-70 kDa and pI 5.0-8.0. The identified proteins include mainly cytoplasmic and nuclear proteins and some mitochondrial, endoplasmic reticulum, and membrane proteins. These proteins are classified into different functional groups such as structure and mobility proteins (21.2%), metabolic enzymes (16.9%), protein folding and metabolism proteins (13.6%), signaling proteins (14.4%), heat-shock proteins (7.6%), and other functional proteins (12.7%). While structure and mobility proteins are mostly represented by protein spots with high abundance, signaling proteins are mostly represented by protein spots with relatively low abundance. Such a 2-DE database for RMC, especially with many signaling proteins and phosphoproteins characterized, will provide a valuable resource for comparative proteomics analysis of normal and pathologic conditions affecting MC function or pathologic progress. PMID- 16315180 TI - Recent advances in peptide and peptidomimetic stereoisomer separations by capillary electromigration techniques. AB - As the stereochemistry of peptides determines their physicochemical properties and biological activities, analytical methods able to discriminate between peptide stereoisomers are important especially with regard to pharmaceutical peptides and peptidomimetics. The present review summarizes recent developments in peptide and peptidomimetic stereoisomer separations by capillary electromigration techniques. The majority of separations were performed by CE while only few reports have been published on the subject of electrochromatography. In addition to systematic studies on the applicability of certain buffer additives and the evaluation of specific experimental conditions, there have been attempts to understand the mechanistic aspects of peptide stereoisomer separations as well as to analyze the structure of peptide-CD complexes. PMID- 16315181 TI - Progress in capillary electrophoresis of biomarkers and metabolites between 2002 and 2005. AB - Biomarker discovery and metabolite research is a fast-growing and extremely important domain not only for the early detection of certain diseases but also for controlling its progress as well as in pharmaceutical investigations. For the analytical separation and identification, CE plays an indisputable role. Capillary systems enhancing different selectivity are applied and connected to different kind of detection systems. As the choice of buffer and its composition is responsible for a successful separation, special emphasis is put on solvent effects in this review. Altogether the most important capillary electrophoretic techniques applied for biomarker and metabolites analysis published between 2002 and 2005 are summarized and discussed. PMID- 16315182 TI - Recent applications of affinity interactions in capillary electrophoresis. AB - Systems biology depends on a comprehensive assignment and characterization of the interactions of proteins and polypeptides (functional proteomics) and of other classes of biomolecules in a given organism. High-capacity screening methods are in place for ligand capture and interaction screening, but a detailed dynamic characterization of molecular interactions under physiological conditions in efficiently separated mixtures with minimal sample consumption is presently provided only by electrophoretic interaction analysis in capillaries, affinity CE (ACE). This has been realized in different fields of biology and analytical chemistry, and the resulting advances and uses of ACE during the last 2.5 years are covered in this review. Dealing with anything from small divalent metal ions to large supramolecular assemblies, the applications of ACE span from low affinity binding of broad specificity being exploited in optimizing selectivity, e.g., in enantiomer analysis to miniaturized affinity technologies, e.g., for fast processing immunoassay. Also, approaches that provide detailed quantitative characterization of analyte-ligand interaction for drug, immunoassay, and aptamer development are increasingly important, but various approaches to ACE are more and more generally applied in biological research. In addition, the present overview emphasizes that distinct challenges regarding sensitivity, parallel processing, information-rich detection, interfacing with MS, analyte recovery, and preparative capabilities remain. This will be addressed by future technological improvements that will ensure continuing new applications of ACE in the years to come. PMID- 16315183 TI - Recent applications of microemulsion electrokinetic chromatography. AB - Compared to MEKC, the presence of a water-immiscible oil phase in the microemulsion droplets of microemulsion EKC (MEEKC) gives rise to some special properties, such as enhanced solubilization capacity and enlarged migration window, which could allow for the improved separation of various hydrophobic and hydrophilic compounds, with reduced sample pretreatment steps, unique selectivities and/or higher efficiencies. Typically, stable and optically clear oil-in-water microemulsions containing a surfactant (SDS), oil (octane or heptane), and cosurfactant (1-butanol) in phosphate buffer are employed as separation media in conventional MEEKC. However, in recent years, the applicability of reverse MEEKC (water-in-oil microemulsions) has also been demonstrated, such as for the enhanced separation of highly hydrophobic substances. Also, during the past few years, the development and application of MEEKC for the separation of chiral molecules has been expanded, based on the use of enantioselective microemulsions that contained a chiral surfactant or chiral alcohol. On the other hand, the application of MEEKC for the characterization of the lipophilicity of chemical substances remains an active and important area of research, such as the use of multiplex MEEKC for the high-throughput determination of partition coefficients (log P values) of pharmaceutical compounds. In this review, recent applications of MEEKC (covering the period from 2003 to 2005) are reported. Emphases are placed on the discussion of MEEKC in the separation of chiral molecules and highly hydrophobic substances, as well as in the determination of partition coefficients, followed by a survey of recent applications of MEEKC in the analysis of pharmaceuticals, cosmetics and health care products, biological and environmental compounds, plant materials, and foods. PMID- 16315184 TI - Interaction of DNA/polycation complexes with phospholipids: stabilizing strategy for gene delivery. AB - The interaction between negatively charged lipid vesicles and positively charged DNA/polylysine complexes was studied. The interaction does not lead to release of DNA from the initial complexes. The particles formed are easy to prepare, they have slight negative charge, small dimensions and show good stability in physiological NaCl solution. Such properties might indicate that stabilization of the particles by lipid coating might be a potent strategy, alternative to PEGylation of DNA/polycation complexes. Interaction of DNA/polycation complex particles with lipid vesicles. PMID- 16315185 TI - Surface properties of silk fibroin films and their interaction with fibroblasts. AB - There is a growing interest in the use of silk as a biomaterial for tissue engineering. Silk threads from Bombyx mori have a fibrous core of fibroin, the protein responsible for biocompatibility and bioactivity, which is surrounded by a family of "gummy" proteins, called sericins, which are almost completely removed during silk degumming. Three different methanol treatments on regenerated fibroin films were used to convert viscous solutions of Silk I to an insoluble crystalline form (Silk II), in an attempt to devise new processing protocols for the creation of a cell guiding fibroin surface. Human fibroblasts (MRC5 line) were used as probes of the cell-biomaterial interaction in the early stages of the process (1 h, 3 h, 6 h and 4 d after seeding). The effect of each treatment on cell adhesion, spreading and distribution was monitored by scanning electron microscopy (SEM) and was correlated to superficial properties (like roughness and crystallinity) and fibroin conformation by means of atomic force microscopy (AFM), used in both topographical and acoustic mode, and attenuated total internal reflection infrared spectroscopy (FTIR-ATR). It was found that traditional methanol treatments where fibroin films were soaked in methanol solution produced roughness patterns that affected only the very early stages of fibroblast adhesion (until 3 h from seeding), while the new treatment proposed could really dialogue with the cells. Its non-homogeneous surface can explain the existence of cells spreading in specific directions and the presence of cell repellent areas even 4 d after seeding. PMID- 16315186 TI - Effects of processing on the structure of zein/oleic Acid films investigated by x ray diffraction. AB - Zein films plasticized with oleic acid were formed by solution casting, by the stretching of moldable resins, and by blown film extrusion. The effects of the forming process on film structure were investigated by X-ray diffraction. Wide angle X-ray scattering (WAXS) patterns showed d-spacings at 4.5 and 10 A, which were attributed to the zein alpha-helix backbone and inter-helix packing, respectively. The 4.5 A d-spacing remained stable under processing while the 10 A d-spacing varied with processing treatment. Small-angle X-ray scattering (SAXS) detected a long-range periodicity for the formed films but not for unprocessed zein, which suggests that the forming process-promoted film structure development is possibly aided by oleic acid. The SAXS d-spacing varied among the samples (130 238 A) according to zein origin and film-forming method. X-ray scattering data suggest that the zein molecular structure resists processing but the zein supramolecular arrangements in the formed films are dependent on processing methods. Structural model for a zein molecular aggregate (based on Matsushima et al.10). Rectangular prisms of individual zein molecules are hexagonally aligned parallel to each other. PMID- 16315187 TI - Glycosidation of cu,zn-superoxide dismutase with end-group aminated dextran: pharmacological and pharmacokinetics properties. AB - Bovine Cu,Zn-SOD was chemically modified with an end-group aminated dextran derivative using a water-soluble carbodiimide as coupling agent. The enzyme retained 81% of the initial catalytic activity after the attachment of about 4.4 mol of polymer per protein subunit. The anti-inflammatory activity of the SOD was two times increased after conjugation with dextran. The modified enzyme was remarkably more resistant to inactivation by H(2)O(2) and its plasma half-life time was prolonged from 4 min to 3.2 h. PMID- 16315188 TI - The role of astragaloside in regeneration of the peripheral nerve system. AB - The aim of this study was to evaluate peripheral nerve regeneration across a 15 mm gap in the sciatic nerve of the rat, using a silicone rubber nerve guide filled with different concentrations of astragaloside (0, 50, 100, and 200 microM). Collagen was also filled in the chambers to prevent the astragaloside from leakage. At the end of 8 weeks, animals from the group treated with astragaloside, especially at the concentration of 50 microM, had a higher rate of successful regeneration across the wide gap, a significantly larger number of myelinated axons, and a greater evoked action potential than the control group. However, the high-dose astragaloside (200 microM) completely reversed this positive effect of growth-promoting capability and inhibited nerve regeneration. Thus, astragaloside plays a dual role in anastomosis, being salutary in aiding the growth of axons in peripheral nerve but also detrimental, terminating the nerve regenerative processes if improperly applied. PMID- 16315189 TI - Investigation into the erosion mechanism of salicylate-based poly(anhydride esters). AB - Salicylate-based poly(anhydride-esters), collectively referred to as PolyAspirin, hydrolytically degrade into salicylic acid, a nonsteroidal anti-inflammatory drug (NSAID). The variations of the poly(anhydride-ester) investigated in this article are linked by adipic acid, suberic acid, or sebacic acid. To elucidate the erosion mechanism of these polymers, water uptake, mass loss, contact angle, and changes in device thickness were monitored as a function of in vitro degradation time. The polymers examined here appeared to primarily undergo surface erosion. The adipate homopolymer absorbed the most water and therefore swelled more than the other versions of the poly(anhydride-ester). Additionally, the adipate homopolymer eroded most quickly. Increasing the length of the linker moiety decreased the driving force for hydrolysis, which prolonged the lifetime of the polymer sample. PMID- 16315190 TI - Biocompatibility and degradation of aliphatic segmented poly(ester amide)s: in vitro and in vivo evaluation. AB - Aliphatic segmented poly(ester amide)s, comprising a crystallizable amide phase and a flexible amorphous ester phase, were investigated for potential use in biomedical applications. By varying the amide content and the type of crystallizable amide segments, the polymer's thermal and mechanical properties can readily be tuned. Polymers with 25 and 50 mol % of amide content are noncytotoxic and sustain growth of fibroblasts onto polymer films. The in vitro degradation of these polymers was followed in PBS (pH 7.4) at 37 degrees C up to 7 months. The poly(ester amide)s showed the characteristics of bulk degradation with a gradual decrease in molecular weight and almost no mass loss. The in vivo degradation of these polymers, followed by subcutaneous implantation in rats up to 6 weeks, was slow and similar to the in vitro degradation. The tissue response upon implantation was followed over 6 weeks. A mild foreign-body reaction, characterized by the presence of macrophages, and sporadically a lymphocyte, were observed in the first week of implantation. After 6 weeks the implant site is characterized by fibrous encapsulation with no signs of inflammation. The poly(ester amide)s tested are biocompatible, but their in vitro as well as in vivo degradation is very slow. PMID- 16315191 TI - Chemical modification of SWNT alters in vitro cell-SWNT interactions. AB - Single-walled carbon nanotubes (SWNT) have been the focus of considerable attention as a material with extraordinary mechanical and electrical properties. SWNT have been proposed in a number of biomedical applications, including neural, bone, and dental tissue engineering. In these applications, it is clear that surrounding tissues will come into surface contact with SWNT composites, and compatibility between SWNT and host cells must be addressed. This investigation describes the gross physical and chemical effects of different SWNT preparations on in vitro cell viability and metabolic activity. Three different SWNT preparations were analyzed: as purchased (AP-NT), purified (PUR-NT), and functionalized with glucosamine (GA-NT), over concentrations of 0.001-1.0% (wt/vol). With the exception of the lowest SWNT concentrations, increasing concentrations of SWNT resulted in a decrease of cell viability, which was dependent on SWNT preparation. The metabolic activity of 3T3 cells was also dependent on SWNT preparation and concentration. These investigations have shown that these SWNT preparations have significant effects on in vitro cellular function that cannot be attributed to one factor alone, but are more likely the result of several unfavorable interactions. Effects, such as destabilizing the cell membrane, soluble toxic contaminants, and limitations in mass transfer as the SWNT coalesce into sheets, may all play a role in these interactions. Using comprehensive purification processes and modifying the NT-surface chemistry to introduce functional groups or reduce hydrophobicity or both, these interactions can be significantly improved. PMID- 16315192 TI - Chitosan monomer promotes tissue regeneration on dental pulp wounds. AB - The present study was undertaken to evaluate the applicability of chitosan monomer (D-glucosamine hydrochloride) as a pulp capping medicament. Both in vitro and in vivo experiments were carried out to study the cell metabolism and wound healing mechanisms following the application of chitomonosaccharide. After 3 days of osteoblast culture, alkaline phosphatase (ALP) activity significantly increased in the chitosan group. Reverse transcription polymerase chain reaction analysis revealed that chitosan induced an increase in the expression of ALP mRNA after 3 days and bone morphogenetic protein-2 mRNA after 7 days of osteoblast incubation. Inflammatory cytokine, interleukin (IL)-8, synthesis in fibroblasts was strongly suppressed in the medium supplemented with chitosan monomer. Histopathological effects were evaluated in rat experiments. After 1 day, inflammatory cell infiltrations were observed to be weak when compared with the application of chitosan polymer. After 3 days, a remarkable proliferation of fibroblasts was seen near the applied chitosan monomer. The inflammatory cell infiltration had almost completely disappeared. After 5 days, the fibroblastic proliferation progressed, and some odontoblastic cells appeared at the periphery of the proliferated fibroblasts. These findings indicate that the present study is the first report that chitosan monomer acts as a biocompatibility stable medicament even at the initial stage of wound healing in comparison with the application of chitosan polymer. PMID- 16315193 TI - Rheological and cohesive properties of hyaluronic acid. AB - Hyaluronic acid (HA) is a naturally occurring polysaccharide with unique biomedical applications. We have studied the cohesive and rheological properties of HA of three molecular weights (0.35 x 10(6) -1.80 x 10(6) Da) and found that the cohesive nature of HA was highly dependent on molecular weight and solution concentration. To a first approximation, the cohesive nature of HA in solution correlates with concentration, independent of molecular weight. Several rheological parameters correlated with molecular weight: zero shear viscosity, complex viscosity, and the complex viscosity at the crossover point. The cohesive properties of the HA solutions, measured by dynamic aspiration (Poyer et al, J Cataract Refract Surg 1998;24:1130-1135), were found to decrease as the zero shear viscosity increases. The cohesive properties of HA polymer in solution were found to correlate with the high frequency complex viscosity and high frequency loss modulus independent of molecular weight. PMID- 16315194 TI - Dynamic chemical devices: photoinduced electron transfer and its ion-triggered switching in nanomechanical butterfly-type bis(porphyrin)terpyridines. AB - A series of butterfly-type molecular constructs has been prepared in good yield by using a double Stille coupling synthetic protocol. They are composed of a terpyridine (terpy) scaffold and two wings composed of appended porphyrins that are capable of switching from an extended W geometry to a compact U geometry upon cation coordination of the terpy unit. The porphyrin moieties exist in the constructs either as free bases or they can be sequentially metallated, thus giving rise to wings of different "colours". Stationary and time-resolved emission studies of the HZn, ZnAu and Zn2Au constructs show that the electronic properties are strongly dependent on the geometry. In the extended W conformation an energy-transfer process is seen from the free base to the Zn-metallated porphyrin. In the U conformation in Zn2Au the donor luminescence resulting from the singlet excited state of the Zn wing is strongly, quenched not only due to the heavy atom effect but also due to a fast electron-transfer process to the ground state of the Au wing. Furthermore, the binding of (alpha,omega)-diamine substrates to the Zn(II)-porphyrin sites can also influence the conformation of the system. For the Zn2Zn construct, single-crystal diffraction experiments with synchrotron radiation allowed the structure to be solved by direct methods and fully refined; it shows the expected U conformation. The central Zn atom is six coordinate, whereby the zinc atom is coordinated by the eta3-terpy ligand as well by monodentate and semi-chelating acetate anions. The structure is made rigid by hydrogen bonds involving the aqua ligands on the outer Zn centres and acetate oxygen atoms. The present system thus represents a double-trigger-modulated optomechanical switching device with selective substrate binding for either metal atoms or tailored ligands. Both energy- and electron-transfer processes can be controlled opening a means of improving the on/off ratio in future constructs. PMID- 16315195 TI - Origin of stereoinduction by chiral aminophosphane phosphinite ligands in enantioselective catalysis: asymmetric hydroformylation. AB - The origin of stereoinduction by chiral aminophosphane phosphinite (AMPP) ligands in asymmetric hydroformylation was investigated with a theoretical approach. The roles of the stereogenic center at the aminophosphane phosphorus atom (NP*) and of the chirality of the backbone were analyzed by considering three experimentally tested cases: 1) P-stereogenic yielding high ee, 2) P nonstereogenic yielding low ee, and 3) P-stereogenic yielding low ee. We succeeded in reproducing the experimentally observed trends for the three studied AMPP ligands. Our results indicated that alkene insertion into the rhodium hydride bond is the selectivity-determining step, and not alkene coordination. Additional calculations on model systems revealed that the different nonbonding weak-type interactions of styrene with the substituents of the NP* stereogenic center in an axial position is responsible for stereodifferentiation. The chirality of the AMPP backbone plays a secondary role. The rationalization of the stereochemical outcome is not straightforward, because two competitive equatorial/axial reaction paths, showing opposite asymmetric induction, must be considered. Construction of stereochemical models and evaluation of stereoinduction for novel ligand systems suggested that two prerequisites are required to improve the performance of AMPP-type ligands in asymmetric hydroformylation: 1) combination of stereorecognition and stereohindrance by substituents at the NP* atom, and 2) more rigid backbones. PMID- 16315196 TI - Silica self-assembly and synthesis of microporous and mesoporous silicates. AB - The microstructure of silica in basic aqueous solutions containing organic cations and prepared from monomeric precursors is reviewed and interpreted within the context of classical ideas of self-assembly of molecular aggregates. The solution properties can be understood by using the pseudo-phase separation approach coupled to the acid-base chemistry of silanol groups and the Poisson Boltzmann equation. The silica nanoparticles frequently observed in these systems have a core-shell structure with silica in the core and the organic cations at the shell. Individual particles are observed when the forces between particles are repulsive-as is the case for small cations such as tetramethylammonium or tetrapropylammonium-and extended structures such as M41S materials are formed when the forces are attractive--as is the case for surfactants such as cetyltrimethylammonium. These ideas are useful to understand the evolution of zeolite synthesis gels from nucleation to crystal growth. Although at room temperature the silica and the organic cations are segregated, upon heating the organic cations are embedded within the particles. This transformation signals the onset of structure direction whereby the size and geometry of the organic cation induce changes in the structure of silica that may lead to zeolite nuclei. PMID- 16315197 TI - Controlling the nature of mixed (phthalocyaninato)(porphyrinato) rare-earth(III) double-decker complexes: the effects of nonperipheral alkoxy substitution of the phthalocyanine ligand. AB - The half-sandwich rare-earth complexes [M(III)(acac)(TClPP)] (M = Sm, Eu, Y; TClPP = meso-tetrakis(4-chlorophenyl)porphyrinate; acac = acetylacetonate), generated in situ from [M(acac)3] x n H2O and H2(TClPP), were treated with 1,8,15,22-tetrakis(3-pentyloxy)phthalocyanine [H2{Pc(alpha-OC5H11)4}] (Pc = phthalocyaninate) under reflux in n-octanol to yield both the neutral nonprotonated and protonated (phthalocyaninato)(porphyrinato) rare-earth double decker complexes, [M(III){Pc(alpha-OC5H11)4}(TClPP)] (1-3) and [M(III)H{Pc(alpha OC5H11)4}(TClPP)] (4-6), respectively. In contrast, reaction of [Y(III)(acac)(TClPP)] with 1,4,8,11,15,18,22,25-octakis(1-butyloxy)phthalocyanine [H2Pc(alpha-OC4H9)8] gave only the protonated double-decker complex [Y(III)H{Pc(alpha-OC4H9)8}(TClPP)] (7). These observations clearly show the importance of the number and positions of substituents on the phthalocyanine ligand in controlling the nature of the (phthalocyaninato)(porphyrinato) rare earth double-deckers obtained. In particular, alpha-alkoxylation of the phthalocyanine ligand is found to stabilize the protonated form, a fact supported by molecular-orbital calculations. A combination of mass spectrometry, NMR, UV visible, near-IR, MCD, and IR spectroscopy, and X-ray diffraction analyses, facilitated the differentiation of the newly prepared neutral nonprotonated and protonated double-decker complexes. The crystal structure of the protonated form has been determined for the first time. PMID- 16315198 TI - Phospholipase cleavage of D- and L-chiro-glycosylphosphoinositides asymmetrically incorporated into liposomal membranes. AB - The nature of chiro-inositol-containing inositolphosphoglycans (IPGs), reported to be putative insulin mediators, was studied by examination of the substrate specificities of the phosphatidylinositol-specific phospholipase C (PI-PLC) and the glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) by using a series of synthetic D- and L-chiro-glycosylphosphoinositides. 3-O-alpha-D Glucosaminyl- (3) and -galactosaminyl-2-phosphatidyl-L-chiro-inositol (4), which show the maximum stereochemical similarity to the 6-O-alpha-D glucosaminylphosphatidylinositol pseudodisaccharide motifs of GPI anchors, were synthesized and asymmetrically incorporated into phospholipid bilayers in the form of large unilamellar vesicles (LUVs). Similarly, 2-O-alpha-D-glucosaminyl- (5) and -galactosaminyl-1-phosphatidyl-D-chiro-inositol (6), which differ from the corresponding pseudodisaccharide motif of the GPI anchors only in the axial orientation of the phosphatidyl moiety, were also synthesized and asymmetrically inserted into LUVs. The cleavage of these synthetic molecules in the liposomal constructs by PI-PLC from Bacillus cereus and by GPI-PLD from bovine serum was studied with the use of 6-O-alpha-D-glucosaminylphosphatidylinositol (7) and the conserved GPI anchor structure (8) as positive controls. Although PI-PLC cleaved 3 and 4 with about the same efficiency as 7 and 8, this enzyme did not accept 5 or 6. GPI-PLD accepted both the L-chiro- (3 and 4) and the D-chiro- (5 and 6) glycosylinositolphosphoinositides. Therefore, IPGs containing L-chiro-inositol only are expected to be released from chiro-inositol-containing GPIs if the cleavage is effected by a PI-PLC, whereas GPI-PLD cleavage could result in both L chiro- and D-chiro-inositol-containing IPGs. PMID- 16315199 TI - Unusual high-temperature structural behaviour in ferroelectric Bi2WO6. AB - The crystal structure of Aurivillius phase ferroelectric Bi2WO6 has been studied in detail as a function of temperature by using high-resolution powder neutron diffraction. In agreement with an earlier study, a transition from space group P2(1)ab to B2cb occurs at about 660 degrees C. This transition corresponds to the loss of one octahedral tilt mode within the perovskite-like WO4 layer of the structure. A second, reconstructive, phase transition occurs around 960 degrees C, corresponding to the ferroelectric Curie point; in contrast to previous suggestions, the structure of this high-temperature phase contains layers of stoichiometry WO4, with WO6 octahedra sharing edges and corners, and with the fluorite-like Bi2O2 layers remaining essentially unchanged. This structure is closely related to that of the ambient temperature phase of lanthanide-doped derivatives, for example, Bi0.7Yb1.3WO6 recently reported. This phase-transition behaviour is in stark contrast to that of other members of the Aurivillius family, such as SrBi2Ta2O9 and Bi4Ti3O12, which retain the archetypal Aurivillius connectivity at all temperatures. PMID- 16315200 TI - Mollusk shell formation: a source of new concepts for understanding biomineralization processes. AB - The biological approach to forming crystals is proving to be most surprising. Mollusks build their shells by using a hydrophobic silk gel, very acidic aspartic acid rich proteins, and apparently also an amorphous precursor phase from which the crystals form. All this takes place in a highly structured chitinous framework. Here we present ideas on how these disparate components work together to produce the highly structured pearly nacreous layer of the mollusk shell. PMID- 16315201 TI - A family of ferro- and antiferromagnetically coupled decametallic chromium(III) wheels. AB - The synthesis and crystal structures of a family of decametallic Cr(III) "molecular wheels" are reported, namely [Cr10(OR)20(O2CR')10] [R' = Me, R = Me (1), Et (2); R' = Et, R = Me (3), Et (4); R' = CMe3, R = Me (5), Et (6)]. Magnetic studies on 1-6 reveal a remarkable dependence of the magnetic behaviour on the nature of R. In each pair of complexes with a common carboxylate (R') the nearest neighbour CrCr magnetic exchange coupling is more antiferromagnetic for the ethoxide-bridged (R = Et) cluster than for the methoxide analogue. In complexes 2, 4 and 6 the overall coupling is weakly antiferromagnetic resulting in diamagnetic (S = 0) ground states for the cluster, whilst in 1 and 5 it is weakly ferromagnetic thus resulting in very high-spin ground states. This ground state has been probed directly in the perdeuterated version of 1 ([D]1) by inelastic neutron scattering experiments, and these support the S = 15 ground state expected for ferromagnetic coupling of ten Cr(III) ions, and they also indicate that a single J-value model is inadequate. The ground state of 5 is large but not well defined. The trends in J on changing R are further supported by density functional calculations on 1-6, which are in excellent agreement with experiment. The very large changes in the nature of the ground state between 1 and 2, and 5 and 6 are the result of relatively small changes in J that happen to cross J = 0, hence changing the sign of J. PMID- 16315202 TI - One-armed artificial receptors for the binding of polar tetrapeptides in water: probing the substrate selectivity of a combinatorial receptor library. AB - We have recently developed a new class of one-armed artificial receptors 1 for the binding of the polar tetrapeptide N-Ac-D-Glu-L-Lys-D-Ala-D-Ala-OH (EKAA) 2 in water using a combined combinatorial and statistical approach. We have now further probed the substrate selectivity of this receptor library 1 by screening a second tetrapeptide substrate (3) with the inverse sequence N-Ac-D-Ala-D-Ala-L Lys-D-Glu-OH (AAKE). This "inverse" substrate is also efficiently bound by our receptors, with K(ass) approximately 6000 M(-1) for the best receptors, as determined both by a quantitative on-bead binding assay and by UV and fluorescence titration studies in free solution. Hence, the inverse tetrapeptide 3 is in general bound two to three times less efficiently than the "normal" peptide 2 (K(ass) approximately 17,000 M(-1)), even though the complexation mainly involves long-range electrostatic interactions and both the receptor and substrate are rather flexible. Molecular modeling and ab initio calculations have been used to rationalize the observed substrate selectivity and to analyze the various binding interactions within the complex. PMID- 16315203 TI - Fast perfusion measurements in rat skeletal muscle at rest and during exercise with single-voxel FAIR (flow-sensitive alternating inversion recovery). AB - Non-invasive measurement of perfusion in skeletal muscle by in vivo magnetic resonance remains a challenge due to its low level and the correspondingly low signal-to-noise ratio. To enable accurate, quantitative, and time-resolved perfusion measurements in the leg muscle, a technique with a high sensitivity is required. By combining a flow-sensitive alternating inversion recovery (FAIR) sequence with a single-voxel readout, we have developed a new technique to measure the perfusion in the rat gastrocnemius muscle at rest, yielding an average value of 19.4 +/- 4.8 mL/100 g/min (n = 22). In additional experiments, perfusion changes were elicited by acute ischemia and reperfusion or by exercise induced by electrical, noninvasive muscle stimulation with varying duration and intensity. The perfusion time courses during these manipulations were measured with a temporal resolution of 2.2 min, showing increases in perfusion of a factor of up to 2.5. In a direct comparison, the results agreed closely with values found with microsphere measurements in the same animals. The quantitative and noninvasive method can significantly facilitate the investigation of atherosclerotic diseases and the examination of drug efficacy. PMID- 16315204 TI - Rapid mapping of flow velocity using a new PARSE method. AB - A new method for flow velocity mapping is presented here. Instead of the conventional approach of employing two images (velocity sensitive and control) to generate velocity information, in the new method one determines the velocity directly from a single-shot acquisition by solving an inverse problem. This technique is a variant of single-shot parameter assessment by retrieval from signal encoding (SS-PARSE). The results of simulation and phantom studies show strong agreement with the actual velocities. The prototype method can measure velocities in the range of -50 to 50 cm/s, which is roughly appropriate for future applications in dynamic blood flow measurement in carotid arteries. PMID- 16315205 TI - Localized in vivo isotropic-anisotropic correlation 1H NMR spectroscopy using ultraslow magic angle spinning. AB - In a previous work (1), the susceptibility broadening in the (1)H NMR metabolite spectrum obtained in a live mouse was separated from the isotropic information, which significantly increased the spectral resolution. This was achieved using ultraslow magic angle spinning (MAS) of the animal combined with a modified phase corrected magic angle turning (PHORMAT) pulse sequence. However, PHORMAT cannot be used for spatially selective spectroscopy. This article introduces a modified sequence called localized magic angle turning (LOCMAT) that makes this possible. Proton LOCMAT spectra were obtained from the liver and heart of a live mouse while the animal was spun at a speed of 4 Hz in a 2 Tesla field. It was found that even in this relatively low field, LOCMAT provided isotropic line widths that were a factor of 4-10 times smaller than those obtained in a stationary animal. Furthermore, the susceptibility broadening of the heart metabolites showed unusual features that are not observed in dead animals. The limitations of LOCMAT and possible ways to improve the technique are discussed. It is concluded that in vivo LOCMAT can significantly enhance the utility of NMR spectroscopy for biomedical research. PMID- 16315206 TI - Dual in vivo magnetic resonance evaluation of magnetically labeled mouse embryonic stem cells and cardiac function at 1.5 t. AB - Cell therapy has demonstrated the potential to restore injured myocardium. A reliable in vivo imaging method to localize transplanted cells and monitor their restorative effects will enable a systematic investigation of this therapeutic modality. The dual MRI capability of imaging both magnetically labeled mouse embryonic stem cells (mESC) and their restorative effects on cardiac function in a murine model of acute myocardial infarction is demonstrated. Serial in vivo MR detection of transplanted mESC and monitoring of the mESC-treated myocardium was conducted over a 4-week period using a 1.5 T clinical scanner. During the 4-week duration, the mESC-treated myocardium demonstrated sustained improvement of the left ventricular (LV) ejection fraction and conservation of LV mass. Furthermore, no significant difference of their restorative effects on the cardiac function was created by the magnetic labeling of mESC. Thus, in vivo MRI enables simultaneous detection of transplanted mESC and their therapeutic effect on the injured myocardium. PMID- 16315207 TI - Undersampled elliptical centric view-order for improved spatial resolution in contrast-enhanced MR angiography. AB - Although contrast-enhanced MR angiography (CE-MRA) has been successfully developed into a routine clinical imaging technique, there is still need for improved spatial resolution in a given acquisition time. Undersampled projection reconstruction (PR) techniques maintain spatial resolution with reduced scan times, and the elliptical centric (EC) view order provides high quality arterial phase images without venous contamination. In this work, we present a hybrid elliptical centric-projection reconstruction (EC-PR) technique to provide spatial resolution improvement over standard EC in a given time. The k-space sampling was performed by undersampling the periphery of the k(Y)-k(Z) phase encoding plane of an EC view order in a PR like manner. The sampled views were maintained on a rectilinear grid, and thus reconstructed by standard 3DFT. The non-sampled views were compensated either by zero-filling or performing a 2D homodyne reconstruction. Compared to a fully sampled k-space, the EC-PR sequence acquired in the same scan time provides a resolution improvement of about two, as shown by point spread function analysis and phantom experiments. The hypothesis that EC-PR provides improved resolution while retaining diagnostically adequate SNR was tested in 11 CE-MRA studies of the popliteal and carotid arteries and shown to be true (P < 0.03). PMID- 16315208 TI - Musculoskeletal tumors: use of proton MR spectroscopic imaging for characterization. AB - PURPOSE: To determine the value of multivoxel proton magnetic resonance spectroscopic imaging (MRSI) in distinguishing malignant skeletal tumors from benign tumors and normal bone marrow using the metabolite choline (Cho) as a marker for malignancy. MATERIALS AND METHODS: Pathologic specimens obtained from 13 patients who had undergone wide resection for skeletal tumors underwent evaluation by MRSI at 1.5 T. Coronal T1-weighted gradient-echo sequence obtained for localization purposes (TR/TE = 250/1.8 msec, field of view [FOV] = 18 x 18), and single-slice MRSI (TR/TE = 2000/272 msec, FOV = 18 x 18, 10-mm slice thickness) were performed. Water, lipid, and Cho images were reconstructed from MRSI data. Cho signal was measured in each specimen and expressed relative to background noise level (signal-to-noise ratio [SNR]) where noise was measured between 7.0 and 9.0 ppm. Cho SNRs were compared between areas containing malignant tumor and nonmalignant tissue (benign lesion or normal bone marrow) as determined by histopathology. RESULTS: Specimens included 13 skeletal sarcomas (seven osteosarcomas, three chondrosarcomas, one malignant fibrous histiocytoma, one fibrosarcoma, and one leiomyosarcoma). All specimens included a sample of normal bone marrow and two specimens also contained benign lesions. All sarcomas demonstrated a signal at 3.2 ppm assigned to Cho-containing metabolites in areas of malignancy. Peak Cho SNR was significantly different for areas containing histologically-proven malignancy compared to nonmalignant tissue (9.8 +/- 5.1 vs. 2.7 +/- 1.4, respectively, P < 0.002). CONCLUSION: These preliminary results indicate that MRSI at 1.5 T is a promising noninvasive method of differentiating malignant skeletal tumors from nonmalignant tissue. Using MRSI, Cho can be detected in skeletal tumors and may serve as a marker for malignancy. PMID- 16315209 TI - Detection of hepatocellular carcinoma by ferumoxides-enhanced MR imaging in cirrhosis: Incremental value of dynamic gadolinium-enhancement. AB - PURPOSE: To investigate the incremental value of dynamic gadolinium-enhancement performed immediately after ferumoxides-enhanced magnetic resonance (MR) imaging on the detection of hepatocellular carcinoma in patients with cirrhosis. MATERIALS AND METHODS: We retrospectively reviewed MR scans of 62 cirrhotic patients over a two-year period. Sequences included ferumoxides-enhanced T2 weighted fast spin echo followed by dynamic gadolinium-enhanced T1-weighted spoiled gradient echo. Two readers independently documented the presence of hepatocellular carcinoma on a three-point confidence scale, without and with gadolinium-enhanced images. The presence or absence of hepatocellular carcinoma was established by histopathology (58 patients) or follow-up imaging (four patients) over a mean period of nine months. RESULTS: A total of 71 hepatocellular carcinomas were found in 42 patients. There was no statistically significant difference in sensitivity for the diagnosis of hepatocellular carcinoma without vs. with gadolinium-enhanced images (68% vs. 74% for reader 1 and 62% vs. 73% for reader 2, respectively, P > 1.3). However, both readers showed a lower mean confidence for tumor detection without vs. with gadolinium enhanced images (2.3 vs. 2.7 for reader 1, 2.3 vs. 2.9 for reader 2, P < 0.01). CONCLUSION: In our study, the addition of dynamic gadolinium-enhancement to ferumoxides-enhanced MR imaging did not improve hepatocellular carcinoma detection, but the addition of gadolinium-enhancement is recommended if ferumoxides-enhanced imaging is used because it increased reader confidence. PMID- 16315210 TI - Anterior-inferior labral lesions of recurrent shoulder dislocation evaluated by MR arthrography in an adduction internal rotation (ADIR) position. AB - PURPOSE: To introduce and evaluate the usefulness of the adduction internal rotation (ADIR) position in MR arthrography for discriminating the following subtypes of Bankart lesions: classic Bankart lesions, anterior labroligamentous periosteal sleeve avulsion (ALPSA) lesions, and Perthes lesions. MATERIALS AND METHODS: The study group consisted of 25 patients who had been referred for MR arthrography of the shoulder and then underwent arthroscopy due to recurrent episodes of shoulder dislocation. MR arthrography was performed in three different positions: neutral, abduction external rotation (ABER), and ADIR. The authors evaluated the statistical significance of the method's discriminative diagnostic ability for the subtypes of Bankart lesions according to the position changes. RESULTS: Twenty-three patients had the following single or complex labral lesions: classic Bankart (N = 8), ALPSA (N = 3), complex classic Bankart/ALPSA (N = 8), and complex ALPSA/Perthes (N = 4). Two patients had no visible anteroinferior labral lesions. The performance of ADIR positioning for differentiating ALPSA lesions was superior to the neutral or ABER positioning. The difference of the discriminative lesion detection ability was statistically significant (P <.01) only for detecting ALPSA lesions in the ADIR position. CONCLUSION: MR arthrography in the ADIR position provides high accuracy for the diagnosis of ALPSA lesions, and complements routine MR arthrography when used to diagnose labroligamentous lesions in patients with recurrent shoulder dislocations. PMID- 16315211 TI - Relaxation times of breast tissue at 1.5T and 3T measured using IDEAL. AB - PURPOSE: To accurately measure T1 and T2 of breast fibroglandular tissue and fat at 1.5T and 3T, and note the partial volume effects of the admixture of fibroglandular tissue and fat on the relaxation rates using an approach termed iterative decomposition of water and fat with echo asymmetry and least squares estimation (IDEAL) imaging. MATERIALS AND METHODS: T1 and T2 values were measured on the right breasts of five healthy women at 1.5T and 3T. T1 data were collected using two sequences: inversion recovery without IDEAL, and inversion recovery with IDEAL. T2 data were collected using Hahn Echo scans. SNR and CNR analyses were conducted on collected data. RESULTS: T1 increased for both fat (21%) and glandular tissue (17%) from 1.5T to 3T. Thus, the TR and TI of breast protocols at 3T should be lengthened accordingly. SNR more than doubled for both tissue types from 1.5T to 3T. IDEAL imaging demonstrated the partial volume effects of fat and glandular tissue on measuring relaxation rates of independent tissue types. CONCLUSION: With separated fat and water images, more precise measurements can be made for the lipid component in fat, and the water component in fibroglandular tissue. PMID- 16315212 TI - Fat-suppressed three-dimensional dual echo Dixon technique for contrast agent enhanced MRI. AB - PURPOSE: To develop a fast T1-weighted, fat-suppressed three-dimensional dual echo Dixon technique and to demonstrate its use in contrast agent enhanced MRI. MATERIALS AND METHODS: A product fast three-dimensional gradient echo pulse sequence was modified to acquire dual echoes after each RF excitation with water and fat signals in-phase (IP) and opposed-phase (OP), respectively. An on-line reconstruction algorithm was implemented to automatically generate separate water and fat images. The signal to noise ratio (SNR) of the new technique was compared to that of the product technique in phantom. In vivo abdomen and breast images of cancer patients were acquired at 1.5 Tesla using both techniques before and after intravenous administration of gadolinium contrast agent. RESULTS: In phantom, the new technique yields a close to the theoretically predicted 41% increase in SNR in comparison to the product technique without fat suppression (FS). In vivo images of the new technique show noticeably improved FS and image quality in comparison to the images acquired of the same patients using the product technique with FS. CONCLUSION: The three-dimensional dual echo Dixon technique provides excellent image quality and can be used for T1-weighted, fat-suppressed imaging with contrast agent injection. PMID- 16315213 TI - Zero filled partial fourier phase contrast MR imaging: in vitro and in vivo assessment. AB - PURPOSE: To validate partial Fourier phase contrast magnetic resonance (PC MR) with full number of excitation (NEX) PC MR measurements in vitro and in vivo. MATERIALS AND METHODS: MR flow measurements were performed using a partial Fourier and a full NEX PC MR sequence in a flow phantom and in 10 popliteal and renal arteries of 10 different healthy volunteers. Average velocity, peak velocity, and flow results were calculated and compared with regression analysis. RESULTS: Excellent correlations in average velocities (r = 0.99, P < 0.001), peak velocities (r = 0.99, P < 0.001), and flow rates (r = 0.98, P < 0.001) were demonstrated in vitro between the two different acquisitions. For the popliteal arteries there was excellent correlation between peak velocities for both acquisitions (r = 0.98, P < 0.0001); the correlation of average velocity measurements when using all data points in the cardiac cycle for all volunteers was 0.96 (P < 0.001). For the renal arteries the same comparison resulted in a good correlation for average velocity (0.93, P < 0.001) and peak velocity measurements (r = 0.91, P = 0.002), although the correlation coefficient for flow rates was 0.88 (P = 0.004). Blurring of the vessel margins was consistently observed on magnitude images acquired with the partial Fourier method, causing overestimation of the vessel area and some error in the flow measurements. CONCLUSION: Partial Fourier PC MR is able to provide comparable average and peak velocity values when using 1 NEX PC MRI as a reference. PMID- 16315214 TI - A case of mesenteric cystic lymphangioma: fat saturation and chemical shift MR imaging. AB - Mesenteric cystic lymphangioma is an uncommon lesion that may be difficult to diagnose radiologically. In our case, a septate cystic mass showed fluid attenuation on a computed tomography scan. We were able to define the fat content within the cystic tumor by magnetic resonance (MR) imaging. The presence of fat within an intra-abdominal cystic mass is suggestive of a dermoid cyst, cystic lymphangioma, or lymphocele. The pathologic diagnosis revealed a cystic lymphangioma, which originated from the mesentery. Although the MR imaging features of dermoid cysts and lymphangiomas are well known, the demonstration of fat content by chemical shift and fat saturation MRI has not yet been reported for a cystic lymphangioma. PMID- 16315215 TI - MRI-compatible incubation chamber for cell culture experiments. AB - PURPOSE: To develop an incubation chamber that is compatible with MRI, while avoiding sources of error due to the experimental setup. MATERIALS AND METHODS: Two identical and gas-tight chambers were constructed of Plexiglas. The temperature and the CO(2) concentration were adjustable. Temperature variations within and between both chambers were measured. The pH values of the cell culture media were measured under different environmental settings. For each environment a colony formation test was carried out. The homogeneity of the magnetic field inside the chambers was estimated by phantom tests. RESULTS: The temperature variations within the chambers were <0.3 degrees C, and the variation between the chambers was on average <0.05 degrees C. After eight hours the pH values of the cell culture media were 7.47 and 7.48 in the reference and measurement chambers, respectively; 7.41 in the CO(2) incubator; and 8.73 in ambient air. In colony formation tests the survival fraction in the Plexiglas chamber was comparable to that in the CO(2) incubator. No distortions of the magnetic field from the incubation chamber were observed. CONCLUSION: The incubation system presented can provide the conditions of a CO(2) incubator without alteration of the magnetic flux density. PMID- 16315216 TI - Muzzle of South American Pleistocene ground sloths (Xenarthra, Tardigrada). AB - Sloths are among the most characteristic elements of the Cainozoic of South America and are represented, during the Pleistocene, by approximately nine genera of gigantic ground sloths (Megatheriidae and Mylodontidae). A few contributions have described their masticatory apparatus, but almost no attention has been paid to the reconstruction of the muzzle, an important feature to consider in relation to food intake, and particularly relevant in sloths because of the edentulous nature of the muzzle and its varied morphology. The relationship between dietary habits and shape and width of the muzzle is well documented in living herbivores and has been considered an important feature for the inference of alimentary styles in fossils, providing an interesting methodological tool that deserves to be considered for xenarthrans. The goal of this study was to examine models of food intake by reconstructing the appearance and shape of the muzzle in five species of Pleistocene ground sloths (Megatherium americanum, Glossotherium robustum, Lestodon armatus, Mylodon darwini, and Scelidotherium leptocephalum) using reconstructions of the nasal cartilages and facial muscles involved in food intake. The preservation of the nasal septum, and the scars for muscular attachment in the rostral part of the skulls, allow making a conservative reconstruction of muzzle anatomy in fossil sloths. Wide-muzzled ground sloths (Glossotherium and Lestodon) had a square, nonprehensile upper lip and were mostly bulk-feeders. The lips, coupled with the tongue, were used to pull out grass and herbaceous plants. Narrow-muzzled sloths (Mylodon, Scelidotherium, and Megatherium) had a cone-shaped and prehensile lip and were mixed or selective feeders. The prehensile lip was used to select particular plants or plant parts. PMID- 16315217 TI - Ultrastructural organization of the anal organs in the anal capsule of Craterostigmus tasmanianus Pocock, 1902 (Chilopoda, Craterostigmomorpha). AB - We describe the ultrastructural organization of the anal organs of Craterostigmus tasmanianus, which are located on the ventral side of the bivalvular anal capsule. Each part of the capsule bears four pore fields with several anal pores. The pores lead into a pore canal, which is surrounded by the single-layered epithelium of the anal organs. Each anal organ is composed of four different cell types: transporting cells of the main epithelium, junctional cells, isolated epidermal glands, and the cells forming the pore canal. The transporting cells exhibit infoldings of the outer cell membranes, forming a basal labyrinth and a poorly developed apical complex. The cells are covered by a specialized cuticle with a widened subcuticular layer. Only the cuticle of the main epithelium is covered by a mucous layer, secreted by the epidermal glands. The ultrastructural organization of the anal organ is comparable to the coxal and anal organs of other pleurostigmophoran Chilopoda. It is likely that the coxal and anal organs of the Pleurostigmophora are homologous, due to their identical ultrastructural organization. Differences concerning the location on the trunk of Pleurostigmophora are not sufficient to reject a hypothesis of homology. Anal organs are found not only in Craterostigmomorpha, but also in most adult Geophilomorpha, and in larvae and most adults of Lithobiomorpha. The anal organs of C. tasmanianus are thought to play an important role in the uptake of atmospheric water. J. Morphol. PMID- 16315218 TI - Quantitative determination of amorphous cyclosporine in crystalline cyclosporine samples by Fourier transform infrared spectroscopy. AB - The purpose of this study is the development of a quantification method to detect the amount of amorphous cyclosporine using Fourier transform infrared (FTIR) spectroscopy. The mixing of different percentages of crystalline cyclosporine with amorphous cyclosporine was used to obtain a set of standards, composed of cyclosporine samples characterized by different percentages of amorphous cyclosporine. Using a wavelength range of 450-4,000 cm(-1), FTIR spectra were obtained from samples in potassium bromide pellets and then a partial least squares (PLS) model was exploited to correlate the features of the FTIR spectra with the percentage of amorphous cyclosporine in the samples. This model gave a standard error of estimate (SEE) of 0.3562, with an r value of 0.9971 and a standard error of prediction (SEP) of 0.4168, which derives from the cross validation function used to check the precision of the model. Statistical values reveal the applicability of the method to the quantitative determination of amorphous cyclosporine in crystalline cyclosporine samples. PMID- 16315219 TI - A mathematical model to predict the size of the pellets formed in freeze pelletization techniques: parameters affecting pellet size. AB - A mathematical model was developed based on the theory of drop formation to predict the size of the pellets formed in the freeze pelletization process. Further the model was validated by studying the effect of various parameters on the pellet size such as viscosity of the pellet forming and column liquids, surface/interfacial tension, density difference between pellet forming and column liquids; size, shape, and material of construction of the needle tips and temperatures maintained in the columns. In this study, pellets were prepared from different matrices including polyethylene glycols and waxes. The column liquids studied were silicone oils and aqueous glycerol solutions. The surface/interfacial tension, density difference between pellet forming and column liquids and needle tip size were found to be the most important factors affecting pellet size. The viscosity of the column liquid was not found to significantly affect the size of the pellets. The size of the pellets was also not affected by the pellet forming liquids of low viscosities. An increase in the initial column temperature slightly decreased the pellet size. The mathematical model developed was found to successfully predict the size of the pellets with an average error of 3.32% for different matrices that were studied. PMID- 16315221 TI - The binding interaction of synthetic ozonide antimalarials with natural and modified beta-cyclodextrins. AB - The current studies were undertaken to explore the potential basis for a significant difference in the pharmacokinetic parameters after intravenous administration of a synthetic ozonide (OZ) antimalarial drug candidate (1) to rats when formulated in either Captisol (a sulfobutylether substituted beta cyclodextrin derivative ((SBE)(7)-beta-CD)) or a buffered aqueous vehicle. It was suspected that the differences may have been due to failure of 1 to rapidly dissociate from the cyclodextrin complex in vivo, perhaps due to an unusually tight binding within the cyclodextrin cavity. To address this hypothesis, the binding of representative synthetic OZ antimalarial drug candidates (including 1) with beta-cyclodextrin and (SBE)(7)-beta-CD was investigated by isothermal titration calorimetry and phase solubility analysis. It was found that each of the OZ compounds exhibited an exceptionally high binding constant ( approximately 10(6)/M) with both Cyclodextrins (CD). The nature of the complexation was investigated by molecular dynamics simulations and NMR to explore the mechanisms, which generated such high binding constants. The data suggested that the most probable cause of the unusually high binding constants was a very close fit within the cyclodextrin cavity that resulted in more favourable changes in both the enthalpy and entropy of the binding interaction, compared to published data for other drugs. PMID- 16315222 TI - Active dimer of Epratuzumab provides insight into the complex nature of an antibody aggregate. AB - Understanding the intermolecular products of antibodies as a consequence of host cell expression, aging, and heat-stress can be insightful especially when it involves the development of a stable biopharmaceutical product. The dimerized form of Epratuzumab (an IgG(1) antibody) with a molecular mass of approximately 300 kDa (twice the monomer antibody molecular weight of approximately 150 kDa) was examined to gain a better perspective of its properties pertaining to structure and activity. The nascent dimer was shown to partially dissociate upon incubation at 30 degrees C and 37 degrees C, exhibit no discernable alteration of structure (i.e., secondary or tertiary structure based on CD and 2nd derivative UV spectroscopy), have approximately 70% covalent forms (based upon CE-SDS results) and manifest twofold higher activity relative to the active monomer form (on a weight basis the dimer and monomer have equal activity). Interestingly, these properties were not attributed to a single dimer species, but rather to a more complex dimer assembly. The Epratuzumab dimer was digested with papain to reveal three uniquely dimerized aggregates. The relative molar distribution of Fab:Fab, Fc:Fc, and Fab:Fc was found to be 4:3:8, respectively. The data suggest that all three predominantly covalent dimer adducts are capable of full activity, shedding light on their complex nature and showing that their target specificity was unaltered. ESI-MS data indicated the presence of remnant levels of noncovalent dimers for all three dimerized forms. Material aged at 37 degrees C exhibited a similar papain digest molar distribution of the three dimerized forms, except with enhanced chemical heterogeneity and an increase in covalent forms to approximately 84%. PMID- 16315223 TI - Relative bioavailability estimation of carbamazepine crystal forms using an artificial stomach-duodenum model. AB - The in vitro dissolution of carbamazepine (CBZ) was investigated using an automated artificial stomach-duodenum (ASD) model. Successful simulation of the dog physiology in the fasted state showed that the rank order of the ASD estimated bioavailabilities is as follows: Form III > Form I > dihydrate. This result is in excellent agreement with those found in literature. Additional simulations comparing different gastric transit times during fasted and fed states are also discussed. PMID- 16315224 TI - Injectable formulation of disodium 1-[2-(carboxylato)pyrrolidin-1-yl]diazen-1-ium 1,2-diolate (PROLI/NO), an ultrafast nitric oxide donor prodrug. AB - PROLI/NO is an agent of structure XN(O)==NONa (X = L-prolyl) whose 2-s half-life for nitric oxide (NO) release at physiological pH makes it an excellent prodrug for localizing NO's therapeutic effects at the site of application, but a difficult one to formulate and certify as pure. Despite its extraordinary thermal and hydrolytic instability, however, PROLI/NO could be formulated as an injectable drug by dissolving it in cold 0.1 M sodium hydroxide containing 5% D mannitol, then quickly ultrafiltering and lyophilizing it in evacuated septum vials. No evidence for decomposition was seen in the contents of these evacuated vials when stored at -20 degrees C over a 140-day observation period, as judged by quantifying NO release in simulated infusate solutions (10 mM carbonate/bicarbonate, pH 10.5). The only hydrolysis products detected were NO, nitrite ion, proline, and N-nitrosoproline, all products of normal human physiological processes. PMID- 16315225 TI - Predicting the aerosol performance of dry powder inhalation formulations by interparticulate interaction analysis using inverse gas chromatography. AB - Previous studies have demonstrated the utility of inverse gas chromatography (IGC) in discriminating the differences in surface energy between salmeterol xinafoate (SX) powders prepared by conventional sequential batch crystallization and micronization and by supercritical fluid crystallization. In the present study, solubility parameters derived from IGC analysis at infinite dilution (zero coverage) were further utilized to evaluate the influence of solid-solid interactions on the in vitro aerosol performance of these SX samples, with or without the inclusion of a lactose carrier. To this end, the strength of cohesive SX-SX interactions and that of adhesive SX-lactose interactions were computed for the samples from the corresponding solubility parameters, and their fine particle fractions determined using a multi-stage liquid impinger. It was found that the aerosol performance of SX could be substantially improved by the addition of lactose carrier only if the adhesive SX-lactose interactions were stronger than the cohesive SX-SX interactions. The difference in strength between these two forms of interactions also displayed a significant correlation with the increase in fine particle fraction after the addition of lactose carrier. These results suggest that IGC-based interparticulate interaction measurements may serve as a useful means for predicting the aerosol performance of dry powder inhalation formulations. PMID- 16315227 TI - Requirements for room temperature shimming of the human brain. AB - Room temperature (RT) shims are used routinely in MRI to remove global and local B(0) field inhomogeneity introduced by the subject. Most clinical scanners use only second-order spherical harmonic terms, but with the increasing availability of very high field systems, third- and fourth-order terms are a serious consideration. However, choosing appropriate coil strengths is of critical importance in shim coil design since overspecification of the shim strengths can lead to a variety of problems, including shim coil self-resonance. In this study B(0) field map data collected over a period of 6 months (over 400 brain volumes) were analyzed to find the characteristic B-fields required to shim these brains. These data can be used to specify the coil requirements to effectively shim the human brain. PMID- 16315228 TI - Pharmaceutical and immunological evaluation of human papillomavirus viruslike particle as an antigen carrier. AB - We report the preparation and the immunogenicity of a conjugate vaccine obtained by chemically conjugating a variant of the extracellular peptide fragment of influenza type A M2 protein to the human papillomavirus (HPV) viruslike particle (VLP). Conjugates comprised of approximately 4,000 copies of the antigenic peptide per VLP are obtained as the result of the reaction between a C-terminal cysteine residue on the peptide and the maleimide-activated HPV VLP. The resulting conjugates have an average particle size slightly larger than the carrier and present enhanced overall stability against chemical and thermal induced denaturation. The M2-HPV VLP conjugates lost the binding affinity for anti-HPV conformational antibodies but retained reactivity to a M2-specific monoclonal antibody. The conjugate vaccine formulated with aluminum adjuvant and delivered in two doses of 30-ng peptide was found to be highly immunogenic and conferred good protection against lethal challenge of influenza virus in mice. These results suggest that HPV VLP can be used as a carrier for synthetic or small antigens for the development of subunit vaccines. PMID- 16315229 TI - Solubilization of two structurally related anticancer drugs: XK-469 and PPA. AB - The efficiency of a solubilization technique is determined by the physical chemical properties of the drug. This study investigates the solubilization on two structurally related anticancer drugs, XK-469 and PPA. XK-469 is much less polar than PPA with an intrinsic solubility of 0.000274 mg/mL, which is about 10,000 fold less than that of PPA. Fortunately, its physical-chemical properties make it much more formulatable. An ionizable drug can be solubilized by pH adjustment with cosolvency, micellization, or complexation. Both XK-469 and PPA are weak acids with pKa values of 2.7 and 2.9, respectively. Thus, they can be solubilized by pH adjustment. At pH 4.55, neither cosolvency, micellization nor complexation has much effect on the solubility of PPA. However, these techniques can significantly increase the solubility of XK-469. In fact, the solubility of XK-469 in 20% HPbetaCD at pH 4.55 is 5.85 mg/mL, which is more than 20,000 times greater than its intrinsic solubility. With the solubilization descriptors obtained from the experimental data for both unionized and ionized drug species at pH 1.0 and pH 4.55, the solubility of each drug at any pH and excipient concentration can be estimated. Then, a solubilization technique can be chosen for preparing a desired final drug concentration. PMID- 16315230 TI - Anthrax vaccine powder formulations for nasal mucosal delivery. AB - Anthrax remains a serious threat worldwide as a bioterror agent. A second generation anthrax vaccine currently under clinical evaluation consists of a recombinant Protective Antigen (rPA) of Bacillus anthracis. We have previously demonstrated that complete protection against inhalational anthrax can be achieved in a rabbit model, by intranasal delivery of a powder rPA formulation. Here we describe the preformulation and formulation development of such powder formulations. The physical stability of rPA was studied in solution as a function of pH and temperature using circular dichroism (CD), and UV-visible absorption and fluorescence spectroscopies. Extensive aggregation of rPA was observed at physiological temperatures. An empirical phase diagram, constructed using a combination of CD and fluorescence data, suggests that rPA is most thermally stable within the pH range of 6-8. To identify potential stabilizers, a library of GRAS excipients was screened using an aggregation sensitive turbidity assay, CD, and fluorescence. Based on these stability profiles, spray freeze-dried (SFD) formulations were prepared at pH 7-8 using trehalose as stabilizer and a CpG containing oligonucleotide adjuvant. SFD formulations displayed substantial improvement in storage stability over liquid formulations. In combination with noninvasive intranasal delivery, such powder formulations may offer an attractive approach for mass biodefense immunization. PMID- 16315231 TI - New application of air-drying techniques for studying Ephemeroptera and Plecoptera eggs by scanning electron microscopy. AB - Hexamethyldisilizane (HMDS) and tetramethylsilane are organic compounds that are volatile at ambient temperature and which can therefore be used for air-drying biological samples for SEM studies. The techniques using these compounds provide results that are comparable with those obtained by critical point drying, but which involve a very simple process that saves time and money. Both techniques were applied to SEM studies of Ephemeroptera and Plecoptera eggs in order to assess their suitability as alternative methods to critical point drying for these kinds of biological material. The results show no morphological differences between eggs HMDS air-dried and critical point-dried. PMID- 16315232 TI - Development of a protocol for multiple staining with fluorochromes to assess the functional status of boar spermatozoa. AB - The aim of this study was to design a simple and reliable method for the simultaneous evaluation of the nucleus, the acrosome, and the mitochondrial sheath of boar spermatozoa. Sperm samples coming from healthy and sexually mature Pietrain boars were incubated with two nuclear fluorochromes--bis-benzamide specific for viable cells, and propidium iodide specific for nonviable cells--the fluorochrome Mitotracker Green FM specific for functional mitochondria, and the lectin Trypsin inhibitor from Soybean (SBTI) conjugated with the fluorochrome Alexa Fluor 488 specific for proacrosin. The results obtained from assessing the functional status of the spermatozoa using fluorochromes were compared with the conventional sperm parameters of sperm vitality using the eosin exclusion test (EE test), and sperm motility and morphology using the computer-assisted semen analyzer SCA 2002Produccio. Applying the multiple staining test, it was found that the frequency of viable spermatozoa with intact acrosome and intact mitochondria was not different from the frequency of viable spermatozoa obtained with the EE test, and also correlated positively with the frequency of motile spermatozoa and the frequency of mature spermatozoa. Therefore, this technique is useful to characterize the status of boar spermatozoa by assessing the nuclear, acrosomal, and mitochondrial integrity. Moreover, it provides reliable diagnostic information about the fertility potential of boars. PMID- 16315233 TI - Environmental scanning electron microscopy observation of the ultrastructure of Demodex. AB - In this study, numbers of Demodex of hair follicles and sebaceous glands were prepared and the ultrastructure (especially the mouthparts) of Demodex was observed firstly with environmental scanning electron microscopy (ESEM). The most suitable treatment methods and optimal environmental condition for observing the genus samples were found. The samples were washed with detergent and rinsed with distilled water, and then were taken to the specimen stage, on which there was carbon adhesive tape, using special tools. When the temperature was at 5 degrees C and chamber pressure at 5 mbar respectively, the surface of the samples could be fully imaged without covering water or dehydration. The sample surfaces were plump and clear without postmortem changes and charging artifacts. Detailed information about each part of Demodex was observed by ESEM, and clear three dimensional images were recorded. The mouthparts of D. folliculorum were composed of a complex set of structures, which included a round oral opening, a sharp oral needle, and a special hypostome that looked like a longitudinal spindle in the central position. On the end segment of palpus, there were seven strong palpal claws located on each side of the mouthparts. D. folliculorum had special piercing mouthparts, while the mouthparts of D. brevis were a simpler structure. We could not observe the oral needle of D. brevis, and there were only five pairs of palpal claws on the end segment of palpus. The offensive organs of Demodex resulted in its pathogenic effects. After studying hundreds of Demodex, we identified both female and male species of D. folliculorum, but only females of D. brevis in our sample. PMID- 16315234 TI - Morphometrical study of plant vacuolar dynamics in single cells using three dimensional reconstruction from optical sections. AB - In higher plants, vacuoles increase their volumes in accordance with cell enlargement and occupy most of the cell volume. However, quantitative analyses of vacuolar contributions during changes in cell morphology have been hampered by the inadequacies and frequent artifacts associated with current three-dimensional (3-D) reconstruction methods of images derived from light microscopy. To overcome the limitations of quantifying 3-D structures, we have introduced 3-D morphometrics into light microscopy, adopting a contour-based approach for which we have developed an interpolation method. Using this software, named REANT, the morphological and morphometrical changes in protoplasts and vacuoles during plasmolysis could be investigated. We employed the tobacco (Nicotiana tabacum) BY 2 cell line No.7, expressing a GFP-AtVam3p fusion protein, BY-GV7, using GFP as a marker of vacuolar membranes (VMs). By vital staining of the plasma membrane (PM) of cells, we simultaneously obtained optical sections of both the PM and VM. We, therefore, reconstructed the 3-D structures of protoplasts and vacuoles before and after plasmolysis. We were able to identify the appearance of elliptical structures of VMs in the vacuolar lumen, and to determine that they were derived from cytoplasmic strands. From the 3-D structures, the volumes and surface areas were measured at the single cell level. The shrinkage of vacuoles accounted for most of the decrease in protoplast volume, while the surface area of the vacuoles remained mostly unchanged. These morphometrical analyses suggest that the elliptical structures are reservoirs for excess VMs that result from the response to rapid decreases in vacuolar and protoplast volumes. PMID- 16315235 TI - Hypoxia-induced differential apoptosis in the central nervous system of the sturgeon (Acipenser shrenckii). AB - Hypoxia is a frequent challenge to aquatic vertebrates as compared with that for their terrestrial counterparts. All vertebrates respond to hypoxia in a similar, but not identical manner, indicating that these responses appeared early in the evolution of vertebrates. The aim of this study is to find out the effects of hypoxia on apoptosis in the central nervous system (CNS) of sturgeon, an archaic fish. With the regional specialization of the CNS, we hypothesize that if cell death does occur, the response will vary between regions, i.e., some CNS areas will be more susceptible to hypoxia than the others would. Sturgeons (Acipenser shrenckii) were subjected to hypoxia by exposure to either air or hypoxic water. After 6- or 30-h recovery they were sacrificed and the following regions of the CNS: retina, olfactory lobe, optic tectum, pituitary, cerebellum, pons/medulla, and spinal cord were examined by the terminal transferase mediated dUTP nick end labeling technique and for the cleaved fragment of activated caspase-3 by Western blotting. In hypoxia-treated sturgeons, the retina, optic tectum, pituitary, and spinal cord were found to have significantly more apoptotic cells than did untreated sturgeons at both 6 and 30 h after the hypoxic insults, indicating prolonged damage. Apoptosis was confirmed by Western blotting of the cleaved fragment of activated caspase-3. Olfactory lobe, cerebellum, and pons/medulla had relatively few apoptotic cells. The CNS of sturgeon showed a differential pattern of apoptosis in response to hypoxia. PMID- 16315236 TI - Two-photon fluorescent microlithography for live-cell imaging. AB - Fluorescent dyes added to UV-cure resins allow the rapid fabrication of fluorescent micropatterns on standard glass coverslips by two-photon optical lithography. We use this lithographic method to tailor fiduciary markers, focal references, and calibration tools, for fluorescence and laser scanning microscopy. Fluorescent microlithography provides spatial landmarks to quantify molecular transport, cell growth and migration, and to compensate for focal drift during time-lapse imaging. We show that the fluorescent patterned microstructures are biocompatible with cultures of mammalian cell lines and hippocampal neurons. Furthermore, the high-relief topology of the lithographed substrates is utilized as a mold for poly(dimethylsiloxane) stamps to create protein patterns by microcontact printing, representing an alternative to the current etching techniques. We present two different applications of such protein patterns for localizing cell adhesion and guidance of neurite outgrowth. PMID- 16315237 TI - Ki67 and doublecortin positive cells in the human prefrontal cortices of normal aging and vascular dementia. AB - Immunohistochemical localizing of the proliferation of Ki67 nuclei and doublecortin positive cells were performed in the prefrontal cortex of normal aged and vascular dementia (multiple infarct dementia) patients. Positive Ki67 nuclei and doublecortin positive cells were observed in both groups, with slightly higher density in the prefrontal cortex of vascular dementia. When the Ki67 sites were superimposed with the neuronal specific enolase localizations, only about 5% of the cells was doubly labeled, indicating few proliferating cells were neurons. This percentage did not vary between specimens of normal aging and those of vascular dementia. PMID- 16315238 TI - GABAB receptor intracellular trafficking after internalization in Paramecium. AB - The number of neurotransmitter receptors on the plasma membrane is regulated by the traffic of intracellular vesicles. Golgi-derived vesicles provide newly synthesized receptors to the cell surface, whereas clathrin-coated vesicles are the initial vehicles for sequestration of surface receptors, which are ultimately degraded or recycled. We have previously shown that GABAB receptors display a punctuate vesicular pattern dispersed on the cell surface and throughout the cytoplasm and are internalized via clathrin-dependent and -independent endocytosis. Here we have studied constitutive GABAB receptor trafficking after internalization in Paramecium primaurelia by confocal laser scanning microscopy and multiple immunofluorescence analysis. After internalization, receptors are targeted to the early endosomes characterized by the molecular markers EEA1 and rab5. Some of these receptors, destined for recycling back to the plasma membrane, traffic from the early endosomes to the endosomal recycling compartment that is characterized by the presence of rab4-immunoreactivity (IR). Receptors that are destined for degradation exit the endosomal pathway at the early endosomes and traffic to the late endosome-lysosome pathway. In fact, some of the GABAB-positive compartments were identified as lysosomal structures by double staining with the lysosomal marker LAMP-1. GABAB vesicle structures also colocalize with TGN38-IR and rab11-IR. TGN38 and rab11 are proteins found in association with post-Golgi and recycling endosomes, respectively. PMID- 16315239 TI - Wavelength and alignment tests for confocal spectral imaging systems. AB - Confocal spectral imaging (CSI) microscope systems now on the market delineate multiple fluorescent proteins, labels, or dyes within biological specimens by performing spectral characterizations. However, we find that some CSI present inconsistent spectral profiles of reference spectra within a particular system as well as between related and unrelated instruments. We also find evidence of instability that, if not diagnosed, could lead to inconsistent data. This variability confirms the need for diagnostic tools to provide a standardized, objective means of characterizing instability, evidence of misalignment, as well as performing calibration and validation functions. Our protocol uses an inexpensive multi-ion discharge lamp (MIDL) that contains Hg+, Ar+, and inorganic fluorophores that emit distinct, stable spectral features, in place of a sample. An MIDL characterization verifies the accuracy and consistency of a CSI system and validates acquisitions of biological samples. We examined a total of 10 CSI systems, all of which displayed spectral inconsistencies, enabling us to identify malfunctioning subsystems. Only one of the 10 instruments met its optimal performance expectations. We have found that using a primary light source that emits an absolute standard "reference spectrum" enabled us to diagnose instrument errors and measure accuracy and reproducibility under normalized conditions. Using this information, a CSI operator can determine whether a CSI system is working optimally and make objective comparisons with the performance of other CSI systems. It is evident that if CSI systems of a similar make and model were standardized to reveal the same spectral profile from a standard light source, then researchers could be confident that real-life experimental findings would be repeatable on any similar system. PMID- 16315240 TI - Modeling truncated hemoglobin vibrational dynamics. AB - We present a study on the near equilibrium dynamics of two small proteins in the family of truncated hemoglobins, developed under the framework of a Gaussian network approach. Effective beta carbon atoms are taken into account besides Calphas for all residues but glycines in the coarse-graining procedure, without leading to an increase in the degrees of freedom (beta Gaussian Model). Normalized covariance matrix and deformation along slowest modes with collective character are analyzed, pointing out anticorrelations between functionally relevant sites for the proteins under study. In particular, we underline the functional motions of an extended tunnel-cavity system running inside the protein matrix, which provide a pathway for small ligands binding with the iron in the heme group. We give a rough estimate of the order of magnitude of the relaxation times of the slowest two overdamped modes and compare results with previous studies on globins. PMID- 16315241 TI - The impact of fluor-18-deoxyglucose-positron emission tomography in the management of colorectal liver metastases. AB - Fluor-18-deoxyglucose-positron emission tomography (FDG-PET) has emerged as a promising diagnostic modality in recurrent colorectal carcinoma. Whole-body FDG PET may be an accurate diagnostic modality to determine whether patients with recurrent hepatic disease are suitable candidates for curative resection. Reports on the use of FDG-PET in patients with recurrent colorectal carcinoma are scarce, especially those on colorectal liver metastases. To assess the usefulness of this emerging modality for the selection of patients to undergo resection for colorectal liver metastases, a systematic (meta)-analysis of the current literature was conducted. In the absence of randomized controlled clinical trials, a traditional meta-analysis could not be performed. An alternative strategy was designed to evaluate the current literature. After a literature search, an index score was devised to evaluate the articles with regard to the impact of FDG-PET in patients with colorectal liver metastases. The index scored articles on several items and, as such, could be considered an objective approach for the assessment of diagnostic, nonrandomized clinical trials. The proposed index proved to be an independent instrument for judging several research questions and was used systematically to address the sensitivity, specificity, and clinical impact of FDG-PET in patients with colorectal liver metastases. For FDG-PET, the pooled sensitivity and specificity results were 88.0% and 96.1%, respectively, for hepatic disease and 91.5% and 95.4%, respectively, for extrahepatic disease. For the 6 articles that reported the highest scores on the index, the sensitivity and specificity of FDG-PET for hepatic metastatic disease were 79.9% and 92.3%, respectively, and 91.2% and 98.4%, respectively, for extrahepatic disease, respectively. For computed tomography, the pooled sensitivity and specificity results were 82.7% and 84.1%, respectively, for hepatic lesions and 60.9% and 91.1%, respectively, for extrahepatic lesions. The percentage change in clinical management due to FDG-PET was 31.6% (range, 20.0 58.0%) in the articles that scored above the mean and reported this item. For the 6 highest scoring studies, the percentage change in clinical management was 25.0% (range, 20.0-32.0%). Despite apparent omissions in the literature, the combined sensitivity and specificity of FDG-PET clearly indicated that FDG-PET has added value in the diagnostic workup of patients with colorectal liver metastases. FDG PET can be considered a useful tool in preoperative staging and produced superior results compared with conventional diagnostic modalities, especially for excluding or detecting extrahepatic disease. PMID- 16315242 TI - Phase II study of high-dose chemotherapy before radiation in children with newly diagnosed high-grade astrocytoma: final analysis of Children's Cancer Group Study 9933. AB - BACKGROUND: High-grade astrocytomas (HGA) carry a dismal prognosis and compose nearly 20% of all childhood brain tumors. The role of high-dose chemotherapy (HDCT) in the treatment of HGA remains unclear. METHODS: In a nationwide study, The Children's Cancer Group (CCG) prospectively evaluated 102 children with HGA and postoperative residual disease for efficacy and toxicity of four courses of HDCT before radiotherapy (RT). Patients were randomly assigned to one of three couplets of drugs: carboplatin/etoposide (Regimen A), ifosfamide/etoposide (Regimen B), or cyclophosphamide/etoposide (Regimen C). After HDCT, all patients were to receive local RT followed by lomustine and vincristine. Twenty-six patients were excluded after central neuroradiographic review (n = 8) or pathology review (n = 18). RESULTS: Of 76 evaluable patients (median age, 11.95 yrs; range, 3-20 yrs), 30 patients relapsed during HDCT, and 11 others did not complete HDCT because of toxicity. Nonhematologic serious toxicities were common (29%), and 21% of patients did not receive RT. Objective response rates were not associated with amount of residual disease and did not statistically differ between regimens: 27% (Regimen A), 8% (Regimen B), and 29% (Regimen C). Overall survival (OS) was 24% +/- 5% at 5 years and did not differ between groups. Median time to an event was longest for Regimen A (283 days compared with 83 and 91 days for Regimens B and C, respectively). The five-year, event-free survival (EFS) rate for all patients was 8% +/- 3% and 14% +/- 7% for Regimen A (P = 0.07). CONCLUSIONS: OS and EFS were not affected by histologic grade. Patients who responded to HDCT had a nominally higher survival rate (P = 0.03 for trend). The authors conclude that these commonly used HDCT regimens provide no additional clinical benefit to conventional treatment in HGA, regardless of the amount of measurable residual tumor. PMID- 16315243 TI - Target ability and therapy efficacy of immunoliposomes using a humanized antihepatoma disulfide-stabilized Fv fragment on tumor cells. AB - Recently the use of peptides in bee venom (PBV) for cancer therapy has attracted considerable attention. However, PBV's extensive use is prohibited by its intense hemolytic activity. In this study, the sterically stabilized liposomal PBV (PBV SL) was prepared using soybean phosphatidylcholine, cholesterol, and cholesterol PEG-COOH. The humanized antihepatoma disulfide-stabilized Fv (hdsFv25) was reengineered, expressed, and coupled to sterically stabilized liposomes using the N-hydroxysuccinimide ester method. The hdsFv25-immunoliposomes (SIL [hdscFv25]) were immunoreactive as determined by ELISA assay. PBV-SIL [hdscFv25] can kill SMMC-7721 cells in vitro with higher efficiency than nontargeted liposomes. PBV SIL [hdsFv25] displayed high antitumor activity and resulted in a significant reduction in tumor size compared to nontargeted liposomes and PBV. These results indicated that this strategy should be applicable to applicable in the treatment of other cancers. PMID- 16315244 TI - Reduced tabletability of roller compacted granules as a result of granule size enlargement. AB - The mechanism for the frequently observed "loss of reworkability or tabletability" of dry-granulated (DG) powders was investigated in detail using microcrystalline cellulose (MCC). It was hypothesized that granule size enlargement is the primary mechanism to the phenomenon. Detrimental effects of size enlargement on tabletability of plastic materials are predictable based on the physical model of interparticulate bonding within a tablet. In absence of extensive fracture of particles/granules, larger particles/granules exhibit lower surface area available for bonding thus lower tensile strength when compressed under identical conditions. Size effects were first demonstrated using different grades of MCC powders, both whole and sieved, of different particle size distributions. Regardless grade and sieve fraction, larger particles always resulted in lower tabletability, that is, lower tensile strength at the same compaction pressure. It was subsequently shown that enlargement of granules also reduced powder tabletability regardless grade of MCC. Tabletability of sieved granules after roller compacted for one, two, and four times decreased monotonically with increasing granule size but independent of the total number of roller compaction. Moreover, tabletability of fine granules (44-106 microm) was higher than that of coarse MCC powder (Avicel PH-200). These results suggest that the primary mechanism for reduced tabletabilty of DG granules of MCC is granule size enlargement rather than "work-hardening." PMID- 16315245 TI - Preparation, characterization, and drug release in vitro of chitosan-glycyrrhetic acid nanoparticles. AB - A suitable carrier chitosan (CS) was used to prepare CS-Glycyrrhetic acid (GLA) nanoparticles under very mild conditions by polyelectrolyte complexation. These nanoparticles were well dispersed and stable in aqueous solution, and the physicochemical properties of which were investigated by FT-IR, dynamic light scattering, transmission electron microscope, fluorescence spectra and zeta potential. It was found that only when the weight ratio of CS to GLA was lower than 16.7, could the nanoparticles be formed. The prepared nanoparticles carried a positive charge and had the dried TEM-assessed size in the range from 20 to 30 nm. The mean hydrated diameter, size distribution and zeta potential of the nanoparticles could be controlled by some factors including the weight ratio of CS to GLA, the average molecular weight of CS and the pH value of the medium. It was also found that GLA encapsulation efficiency into the nanoparticles increased with the increase of the weight ratio of CS to GLA. The experiment of GLA release in vitro showed that the effect of CS encapsulation on GLA release was obvious and the CS-GLA nanoparticles system might be used to provide a continuous release. PMID- 16315246 TI - Mediastinal node and diaphragmatic targeting after intracavitary injection of avidin/99mTc-blue-biotin-liposome system. AB - A method for delivering drugs to sites of disease extension in mediastinal nodes is described. Mediastinal node and lymphatic distributions were determined after intracavitary injection of the avidin/biotin-liposome system in normal rats. The effect of the injected dose on lymphatic targeting of liposomes after intraperitoneal injection of (99m)Tc-blue-biotin-liposomes and intrapleural injection of avidin, and vice versa, is presented. Scintigraphic imaging was used to follow the movement of (99m)Tc-blue-biotin-liposomes to determine the pharmacokinetics and organ uptake. Tissue biodistribution studies were performed 22 h after injection of the (99m)Tc-blue-biotin-liposomes. Results indicated that independent of the cavity in which each agent was injected, a dose of 5.0 mg of each agent results in higher mediastinal node targeting (8%-10% ID/Organ) as compared with the injection of a 0.5 mg dose (2%-5% ID/Organ, p < 0.05). Targeting of diaphragm and associated lymphatics was observed when (99m)Tc-blue biotin-liposomes were injected in peritoneum and avidin in pleural space. In contrast, pleural, and pericardial lymphatic targeting was observed when (99m)Tc blue-biotin-liposomes were injected in pleural space and avidin in peritoneum. Intracavitary injection of the avidin/biotin-liposome system could potentially be used for the delivery of prophylactic drugs that could reduce tumor metastasis and infection spread to mediastinal nodes. PMID- 16315247 TI - Sildenafil citrate (Viagra) complexes with bivalent ions. AB - The interaction of Ca(2+)-ions with sildenafil citrate (Viagra) leads to the precipitation of a new polymorph variety of sildenafil base. Under the same conditions, Mg(2+), Zn(2+), and Cd(2+) ions form structurally related crystalline complexes of the composition Me(2+)C(28)H(34)N(6)O(11)S. Lattice parameters have been determined showing that magnesium compound belongs to an orthorhombic system, while the zinc and cadmium compounds are its monoclinic distortions. All three compounds are thermally stable, undergoing decomposition above 175 degrees C with the consequent formation of carbonates Me(2+)CO(3) and oxides. PMID- 16315248 TI - The predictive value of apoptosis protease-activating factor 1 in rectal tumors treated with preoperative, high-dose-rate brachytherapy. AB - BACKGROUND: The objective of this study was to assess the value of apoptosis protease-activating factor 1 (APAF-1) as a predictive marker of response in rectal tumors treated with preoperative, high-dose-rate endorectal brachytherapy. METHODS: Immunohistochemistry for APAF-1 was performed on 94 rectal tumor biopsy specimens from patients who were treated on a preoperative, high-dose-rate brachytherapy protocol. Tumors were considered positive when > 10% of tumor cells were immunoreactive. The association between APAF-1 expression and tumor response was made using the chi-square test. RESULTS: Forty-four tumors (43%) were positive for APAF-1. Thirty tumors had complete pathologic tumor regression after preoperative radiotherapy. Of these, 18 tumors were positive for APAF-1. A partial response occurred in 35 tumors. Eighteen tumors (51%) were positive for the protein. Only 8 of 29 nonresponsive tumors (28%) were immunoreactive for APAF 1. A significant association was found between complete tumor regression and positive APAF-1 status (P = 0.018). APAF-1 expression in partially responsive tumors was significantly greater than in nonresponsive tumors (P = 0.03). CONCLUSIONS: APAF-1 expression in pretreatment rectal tumor biopsy specimens may be useful as a predictive marker of response to preoperative radiotherapy in patients with rectal carcinoma. PMID- 16315249 TI - Rasputin used hypnosis: reply to "Russia's imperial blood". PMID- 16315250 TI - Monoclonal gammmopathy (MG) in extranodal marginal zone lymphoma (ENMZL). PMID- 16315251 TI - Medical care utilization and mortality in sickle cell disease: a population-based study. AB - The purpose of this study was to evaluate the pattern of medical care utilization and mortality in children and adults with sickle cell disease (SCD) in the state of Tennessee. Rates of hospitalization, emergency department visits, and deaths were measured in a cohort of adults and children with SCD enrolled in TennCare, Tennessee's Medicaid managed health care program, from January 1995 to December 2002. TennCare data linked to Tennessee vital records were used to define the population and identify the outcomes. For children less than 5 years of age, the mortality rate was similar to that of other black Tennessee children (P = 0.71). Among children, the death rate was highest in 10-19 years of age and was 8-fold higher than Tennessee's race- and age-specific rate. Among 20- to 49-year-old patients with SCD, mortality was significantly higher in males than in females (P < 0.001). As compared to the black population without SCD in TennCare, patients with SCD had 7-30 times higher rate of hospitalization and 2-6 times higher rates of emergency department visits (P < 0.001). The death rate in adolescents and young adults with SCD continues to be much higher than population-specific rates. Interventions to prevent morbidity and mortality related to SCD are urgently needed. PMID- 16315252 TI - Changes in the natural history of progressive multifocal leukoencephalopathy in HIV-negative lymphoproliferative disorders: impact of novel therapies. AB - The aims of this study were to evaluate the clinical characteristics of HIV negative patients affected by lymphoproliferative disorders (LPD) who developed progressive multifocal leukoencephalopathy (PML), to delineate the risk factors, and to analyze whether the new antineoplastic therapies are changing the natural history of this infectious disease. We retrospectively analyzed 46 cases with confirmed LPD-associated PML published from 1958 to 2004. Patients were stratified according to two different time periods: group A included patients diagnosed before 1989, and group B included patients diagnosed since 1990, after introduction of purine analogues. Group A patients (n = 22) had received alkylating agents and/or radiotherapy, and the majority (63.6%) had advanced Hodgkin disease. At univariate analysis, uncontrolled Hodgkin disease was the only risk factor for PML. In group B patients (n = 24), the most frequent treatments received were purine analogues (58.3%) and high-dose therapy with hematopoietic stem cell transplantation (33.3%; HDT/HSCT). B-cell chronic lymphocytic leukemia (45.8%) and aggressive non-Hodgkin lymphoma (24.9%) were the most frequent underlying LPDs. Patients treated with purine analogues were more likely to have active LPD, lower CD4 cell counts, and to be older and male than were HSCT recipients. The median interval from purine analogues or HDT/HSCT to PML was 11 months. In HDT/HSCT recipients, this interval was delayed for 10 months when peri-transplantation rituximab was used. Univariate analysis identified age >55 years, male sex, and CD4 cell counts 5 cm and < or =8 cm, 2 or 3 lesions at least 1 >3 cm but < or =5 cm with total tumor diameter of < or =8 cm, or 4 or 5 nodules all < or =3 cm with total tumor diameter < or =8 cm. Patients were eligible for living donor liver transplantation (LDLT) if tumors were downstaged to within proposed University of California, San Francisco (UCSF) criteria.13 A minimum follow-up period of 3 months after downstaging was required before cadaveric OLT or LDLT, with imaging studies meeting criteria for successful downstaging. Among the 30 patients enrolled, 21 (70%) met criteria for successful downstaging, including 16 (53%) who had subsequently received OLT (2 with LDLT), and 9 patients (30%) were classified as treatment failures. In the explant of 16 patients who underwent OLT, 7 had complete tumor necrosis, 7 met T2 criteria, but 2 exceeded T2 criteria. No HCC recurrence was observed after a median follow-up of 16 months after OLT. The Kaplan-Meier intention-to-treat survival was 89.3 and 81.8% at 1 and 2 yr, respectively. In conclusion, successful tumor downstaging can be achieved in the majority of carefully selected patients, but longer follow-up is needed to further access the risk of HCC recurrence after OLT. PMID- 16315295 TI - Orthotopic liver transplantation in human immunodeficiency virus (HIV)-positive patients: outcome of 7 patients from the Bonn cohort. AB - The outcome and clinical features of 7 HIV-positive patients who were liver transplanted at Bonn University in the era of highly active antiretroviral therapy (HAART) between 1997 and 2004, analyzed by retrospective chart review, are reported. Reasons for orthotopic liver transplantation (OLT) were end-stage liver disease due to chronic hepatitis C (n = 4) or hepatitis B (n = 1) or acute liver failure due to fulminant hepatitis B (n = 2). Immunosuppression was based on cyclosporine A and prednisone. HAART was reinitiated 1 month after transplantation, and immunosuppression was carefully adapted to account for drug drug interactions between cyclosporine A and protease inihibitors. Prednisone was withdrawn 5 months (median) after OLT when immunosuppression had been reliably established in the presence of HAART. One patient died 95 days after OLT due intrathoracic hemorrhage, whereas 6 patients were alive at a median of 24 months. A single episode of acute rejection was observed. The spectrum of postoperative complications was no different from HIV-negative patients apart from Kaposi's sarcoma and multicentric Castleman's disease in a single patient. Recurrent hepatitis B infection was efficiently prevented, whereas hepatitis C reinfection occurred in all 4 patients who had preexisting hepatitis C. Earlier reports on fatal courses of recurrent hepatitis C infection, high rates of organ rejection, and HAART-related liver toxicity were not observed in our patients. In conclusion, even though preliminary, our data suggest that outcomes after liver transplantation of HIV-infected patients can be improved. PMID- 16315296 TI - Validity of preoperative volumetric analysis of congestion volume in living donor liver transplantation using three-dimensional computed tomography. AB - Reconstruction of middle hepatic vein (MHV) tributaries is controversial in right lobe living donor liver transplantation (LDLT). This study aimed to evaluate the appropriateness of reconstructing MHV tributaries by volumetry using 3 dimensional computed tomography (3D-CT). Between November 2003 and January 2005, 42 donor livers (right-lobe graft, n = 25; left-lobe graft, n = 17) were evaluated using this software. The total congestion volume (CV) associated with the MHV tributaries and the inferior right hepatic vein (IRHV), and graft volume (GV) were calculated. In recipients with right-lobe grafts, CV/(right liver volume [RLV]) and (GV - CV)/(standard liver volume [SLV]) were compared between 2 groups: with reconstruction (n = 16) and without reconstruction (n = 9). To evaluate the influence of CV on the remnant right lobe in donors, total bilirubin was compared between 2 groups: high CV (CV > 20%, n = 13) or low CV (CV < or = 20%, n = 4). The mean CV/RLV ratio was 32.3 +/- 17.1% (V5, 15.2 +/- 9.9%; V8, 9.2 +/- 4.1%; and IRHV, 8.5 +/- 11.4%) and the maximum ratio was as high as 80.8%. The mean (GV - CV)/SLV ratio before reconstruction in patients with or without reconstruction resulted in 33.5 +/- 12.8% and 55.4 +/- 12.9%, respectively (P < 0.01). In donors, total bilirubin was significantly high in the high CV group on postoperative day 1 compared with the low CV group (P < 0.05). In conclusion, calculation of CV using 3D-CT software proved to be very useful. We concluded that this evaluation should be an integral part of procedure planning, especially for right-lobe LDLT. PMID- 16315297 TI - Recurrent hepatitis C after retransplantation: factors affecting graft and patient outcome. AB - Retransplantation (re-LT) of patients with recurrent hepatitis C virus (HCV) carries significant morbidity and mortality, negatively impacting on an already scarce donor allograft pool. In this study, we investigated the outcome of allografts and patients after re-LT due to recurrent HCV. Between 1989 and 2002, 47 patients were retransplanted at our institution due to HCV-related graft failure. Clinical HCV recurrence after re-LT was diagnosed when patients had acute liver enzyme elevation correlated with histological recurrence. The independent influence of these variables on survival was tested using Cox regression model. Chi-squared tests were used to examine the influence of individual demographic and pre/perioperative variables on recurrence. Thirty-one (66%) patients died after re-LT (median 2.2 months). Donor age >60, clinical HCV recurrence, and graft failure due to cirrhosis were significant risk factors for mortality (risk ratios of 3.6, 3.3, and 2.4, respectively). Pre-LT MELD score was lower among survivors (22+/- 5 vs. 27+/- 8). Following re-LT, 38 patients had at least one biopsy due to acute liver dysfunction; 19 of them (50%) had recurrence within the first 3 months. High-dose solumedrol was correlated with early recurrence. No association was found between time of recurrence after the first LT and time of recurrence after re-LT. In conclusion, patients with cirrhosis due to recurrent HCV undergoing re-LT have an extremely high mortality rate; older allografts should be avoided in retransplanting these patients. The timing of clinical recurrence after initial liver transplantation is not predictive of the timing of recurrence after re-LT. Patients experiencing early graft failure due to accelerated forms of HCV should not be denied re-LT with the expectation that a similar disease course will occur after re-LT. PMID- 16315298 TI - Effects of hyperbaric oxygen exposure on experimental hepatic ischemia reperfusion injury: relationship between its timing and neutrophil sequestration. AB - Recent studies have shown that hyperbaric oxygen therapy (HBOT) reduces neutrophil endothelial adherence in venules and also blocks the progressive arteriolar vasoconstriction associated with ischemia-reperfusion (I-R) injury in the extremities and the brain. In order to elucidate the effects of HBOT after I R in digestive organs, particularly in the liver, we evaluated the following: 1) the relationship between timing of HBOT and tissue damage; and 2) HBOT's effects on neutrophil sequestration. Using a hepatic I-R (45 minute) model in male rats, survival rate, liver tissue damage, and neutrophil accumulation within the sinusoids in the HBOT-treated group (Group H) were compared to those in the nontreated group (Group C). For the HBOT-treated group, HBOT was administered as 100% oxygen, at 2.5 atm absolute, for 60 minutes. When HBOT was given 30 minute after I-R, the survival rate was much better in Group H than in Group C. HBOT performed within 3 hours of I-R markedly suppressed increases in the malondialdehyde level in tissues of the liver and lessened the congestion in the sinusoids. In addition, HBOT just after I-R caused decreased number of cells stained by the naphthol AS-D chloroacetate esterase infiltrating into the sinusoids. HBOT 3 hours after reperfusion, however, showed no clear effects upon neutrophil sequestration compared to Group C. These results indicate that HBOT performed within 3 hours of I-R alleviates hepatic dysfunction and improves the survival rate after I-R. Herein, we propose 1 possible mechanism for these beneficial effects: early HBOT given before neutrophil-mediated injury phase may suppress the accumulation of neutrophils after I-R. In conclusion, we believe that the present study should lead to an improved understanding of HBOT's potential role in hepatic surgery. PMID- 16315299 TI - Roles and pitfalls of transplantation in human porphyria. PMID- 16315300 TI - Complications and use of intracranial pressure monitoring in patients with acute liver failure and severe encephalopathy. AB - Monitoring of intracranial pressure (ICP) in acute liver failure (ALF) is controversial as a result of the reported complication risk (approximately 20%) and limited therapeutic options for intracranial hypertension. Using prospectively collected information from 332 patients with ALF and severe encephalopathy, we evaluated a recent experience with ICP monitoring in the 24 centers constituting the U.S. ALF Study Group. Special attention was given to the rate of complications, changes in management, and outcome after liver transplantation (LT). ICP monitoring was used in 92 patients (28% of the cohort), but the frequency of monitoring differed between centers (P < 0.001). ICP monitoring was strongly associated with the indication of LT (P < 0.001). A survey performed in a subset of 58 patients with ICP monitoring revealed intracranial hemorrhage in 10.3% of the cohort, half of the complications being incidental radiological findings. However, intracranial bleeding could have contributed to the demise of 2 patients. In subjects listed for LT, ICP monitoring was associated with a higher proportion of subjects receiving vasopressors and ICP-related medications. The 30-day survival post-LT was similar in both monitored and nonmonitored groups (85% vs. 85%). In conclusion, the risk of intracranial hemorrhage following ICP monitoring may have decreased in the last decade, but major complications are still present. In the absence of ICP monitoring, however, patients listed for LT appear to be treated less aggressively for intracranial hypertension. In view of the high 30-day survival rate after LT, future studies of the impact of intracranial hypertension should also focus on long-term neurological recovery from ALF. PMID- 16315301 TI - A case of successful enteroscopic balloon dilation for late anastomotic stricture of choledochojejunostomy after living donor liver transplantation. AB - Biliary complications remain a major concern after living donor liver transplantation. We describe a pediatric case who underwent a successful endoscopic balloon dilatation of biliary-enteric stricture following living donor liver transplantation using a newly developed method of enteroscopy. The 7-year old boy with late biliary stricture of choledochojejunostomy was admitted 6 years after transplantation. Since percutaneous transhepatic cholangiography was technically difficult in this case, endoscopic retrograde cholangiography was performed using a double-balloon enteroscope under general anesthesia. The enteroscope was advanced retrograde through the duodenum, jejunum, and the leg of Roux-Y by the double-balloon method, and anastomotic stricture of choledochojejunostomy was clearly confirmed by endoscopic retrograde cholangiography and endoscopic direct vision. Balloon dilatation was performed and the anastomosis was expanded. Restenosis was not noted as of 2 years after the treatment. In conclusion, endoscopic balloon dilation of biliary-enteric anastomotic stricture using a new enteroscopic method can be regarded as an alternative choice to percutaneous transhepatic management and surgical re anatomists. PMID- 16315304 TI - High dose intravenous immunoglobulin treatment: mechanisms of action. AB - Intravenous immunoglobulin (IVIg) treatment was introduced as replacement therapy for patients with antibody deficiencies, but evidence suggests that a wide range of immune-mediated conditions could benefit from IVIg. The immunoglobulins are precipitated from human plasma by fractionation methods. In conclusion, the differences in basic fractionation methods and the addition of various modifications for purification, stabilization, and virus inactivation result in products significantly different from each other. PMID- 16315305 TI - Activation pattern of mitogen-activated protein kinases in early phase of different size liver isografts in rats. AB - Mitogen-activated protein kinases (MAPK) play a pivotal role in ischemia reperfusion injuries of heart and liver, but the activation pattern of MAPKs in the early phase of different size liver isografts remains unclear. The experiment is designed to investigate the activation pattern and role of MAPKs in isografts of the rat with different size liver transplantation. The animal models of different size graft liver transplantation (whole graft, 50% size, or 30% size, respectively) were established and the sham operation group served as a control. The recipients were sacrificed at 0.5-, 2-, 6-, and 24-hour time points after transplantation to harvest the graft specimens and blood samples. The serum aspartate amino transferase (AST), alanine amino transferase (ALT) and tumor necrosis factor-alpha (TNF-alpha) levels, and histological findings were evaluated. The expressions of the total and phosphorylated p46/p54 JNKs, p38 MAPK, and p42/p44 ERKs were detected by Western blot. The serum ALT and AST levels increased significantly at the 0.5-hour time point and maintained high with the peak levels at the 6-hour time point after liver transplantation. The different sizes of liver isografts did not change the expressions of total p46/p54JNKs, p38MAPK, and p42/p44 ERKs. While the expressions of phosphorylated p46/p54JNKs, p38 MAPK, and p42/p44 ERKs were either negative or mildly up regulated in the sham operation group, they were significantly activated in the transplanted liver at the 0.5-hour time point, especially in the 30% size liver transplantation group. In conclusion, the activation of three MAPKs in liver isografts correlates with graft size and the JNK and p38 MAPK are responsible for the graft injury while the ERK signal pathway maybe participate in the regulation of cell growth and differentiation after small-for-size liver transplantation. PMID- 16315306 TI - Preincubation of rat and human hepatocytes with cytoprotectants prior to cryopreservation can improve viability and function upon thawing. AB - Cryopreservation of human hepatocytes is important for the treatment of liver disease by hepatocyte transplantation and also for the use of hepatocytes as an in vitro model of the liver. One factor in the success of cryopreservation is the quality of cells before freezing. Preincubation of hepatocytes with cytoprotective compounds to allow recovery from the isolation process prior to cryopreservation, such as those that will boost cellular adenosine triphosphate (ATP) content or antioxidants, may improve the viability and function of cells upon thawing. Rat hepatocytes were used to investigate the effects of preincubation with 10 compounds: precursors (glucose, fructose, glutathione, and S-adenosyl-L-methionine), antioxidants (ascorbic acid and alpha-lipoic acid), and compounds with multiple effects (N-acetylcysteine, pentoxifylline, prostaglandin E(1), and tauroursodeoxycholic acid). Human hepatocytes were then used to investigate 5 of the original 10 compounds (glucose, fructose, alpha-lipoic acid, S-adenosyl-L-methionine, and pentoxifylline). Glucose preincubation (100-300 mM) improved the viability and attachment efficiency of rat hepatocytes and improved the viability and reduced lactate dehydrogenase (LDH) leakage of human hepatocytes. Fructose preincubation (100-300 mM) improved the viability and attachment efficiency of rat hepatocytes and improved the attachment efficiency of human hepatocytes. alpha-lipoic acid preincubation (0.5-5 mM) improved the viability and attachment efficiency of both rat and human hepatocytes. At a concentration of 2.5 mM alpha-lipoic acid also improved the albumin production of human hepatocytes. In conclusion, preincubation of hepatocytes prior to cryopreservation can improve the viability and function of thawed cells and may provide a method of obtaining better-quality cryopreserved hepatocytes for transplantation. PMID- 16315307 TI - The effect of HLA mismatches, shared cross-reactive antigen groups, and shared HLA-DR antigens on the outcome after pediatric liver transplantation. AB - The aim of this study was to analyze the effect of human leukocyte antigen (HLA) class I and HLA-DR mismatching, sharing cross-reactive antigen groups (CREGs), and sharing HLA-DR antigens on the outcome after pediatric liver transplantation. Outcome parameters were graft survival, acute rejection, and portal fibrosis. A distinction was made between full-size (FSLTx) and technical-variant liver transplantation (TVLTx). A total of 136 primary transplants were analyzed. The effect of HLA on the outcome parameters was analyzed by adjusted multivariate logistic and Cox regression analysis. HLA mismatches, shared CREGs, and shared HLA-DR antigens affected neither overall graft survival nor survival after FSLTx. Survival after TVLTx was superior in case of 2 mismatches at the HLA-DR locus compared to 0 or 1 mismatch (P = 0.01) and in case of no shared HLA-DR antigen compared to 1 shared HLA-DR antigen (P = 0.004). The incidence of acute rejection was not influenced by HLA. The incidence of portal fibrosis could be analyzed in 62 1-yr biopsies and was higher after TVLTx than FSLTx (P = 0.04). The incidence of portal fibrosis after TVLTx with 0 or 1 mismatch at the HLA-DR locus was 100% compared to 43% with 2 mismatches (P = 0.004). After multivariate analysis, matching for HLA-DR and matching for TVLTx were independent risk factors for portal fibrosis. In conclusion, an overall beneficial effect of HLA matching, sharing CREGs, or sharing HLA-DR antigens was not observed. A negative effect was present for HLA-DR matching and sharing HLA-DR antigens on survival after TVLTx. HLA-DR matching might be associated with portal fibrosis in these grafts. PMID- 16315308 TI - Hepatic and intestinal schistosomiasis after orthotopic liver transplant. AB - Schistosomiasis affects 200 to 250 million people worldwide. Hepatic schistosomiasis is a well-recognized cause of chronic liver disease and portal hypertension. There are no previous reports of schistosomiasis post liver transplantation. We report on 2 cases of schistosomiasis in liver transplant recipients--a case of gastric schistosomiasis and a case of hepatic schistosomiasis. A discussion of the pathology of schistosomal infection and a rationale for screening potential liver transplant recipients from endemic areas follows. PMID- 16315309 TI - Recurrent hepatic sarcoidosis post-liver transplantation manifesting with severe hypercalcemia: a case report and review of the literature. AB - Sarcoidosis is a systemic granulomatous disease primarily involving the lungs, lymph nodes, skin, eyes and nervous system; liver involvement is asymptomatic in most cases. However, once the patient develops clinical symptoms liver disease is usually progressive and may necessitate orthotopic liver transplantation. There are a few reports of asymptomatic recurrent sarcoidosis developing within the liver allograft. We report a case of early recurrence of sarcoidosis in the liver allograft diagnosed on biopsy in a patient who presented with severe hypercalcemia, kidney dysfunction, and increase in size of abdominal lymph nodes. The liver chemistry tests were within normal limits. The patient responded well to steroid treatment by normalizing serum calcium and creatinine levels and reducing lymph node size. To date, there has been no report in the literature of symptomatic recurrence of hepatic sarcoidosis following orthotopic liver transplantation. PMID- 16315310 TI - Bacterial, mycobacterial, and protozoal infections after liver transplantation- part I. PMID- 16315311 TI - Herpes simplex virus-associated acute liver failure: a difficult diagnosis with a poor prognosis. AB - We report 5 cases of acute liver failure related to herpes simplex (HSV) infection in 1 immunocompetent and 4 immunosuppressed patients. One patient was too ill for liver transplantation indication. Three patients, among the 4 listed, underwent liver transplantation. Three patients died 11 days to 1 year after transplantation and 2 patients died 2 to 3 days after admission. All presented with fever and none with skin lesions. The diagnosis of HSV-related hepatitis was made antemortem in only 2 patients on the basis of positive blood cultures and/or immunohistochemic findings. In the remaining patients, HSV diagnosis was made retrospectively on further histologic and virologic investigations. Primary HSV infection was certain or likely in all cases, including an HSV2 superinfection of an anti-HSV1-positive patient and two HSV superinfections of hepatitis B virus (HBV)-related chronic liver disease. In these latter patients, HSV diagnosis was totally unsuspected, despite fever. HSV superinfection has significantly contributed to liver dysfunction aggravation and death. In conclusion, the diagnosis of HSV hepatitis is difficult to establish in the absence of specific clinical signs. This may suggest the need for early administration of acyclovir in patients with suspected HSV hepatitis, without waiting for virologic confirmation. Diagnosis methods providing fast results (real-time polymerase chain reaction [PCR]) should be implemented. PMID- 16315312 TI - Chicken pox after pediatric liver transplantation. AB - Previous case series have reported serious complications of chicken pox (CP) after pediatric liver transplantation (PLT), mainly due to visceral dissemination. The goal of our study was to determine the incidence, risk factors, and outcomes of CP after PLT. A case-control study of all CP infections in pediatric transplant recipients followed at our center from September 1993 to April 2004 was performed. Data were collected before and after infection and at the same time points in age-, gender-, and transplant year-matched controls. Potential risk factors prior to CP and adverse outcomes after infection were compared between cases and controls. Twenty (6.2%) developed CP at a median of 1.8 yr (0.6-4.8) after PLT. All CP infections were cutaneous, with no evidence of organ involvement. Twelve were hospitalized: 9 only to receive intravenous acyclovir and 3 stayed > or =2 weeks for other complications. Risk factors were not statistically different among cases and controls. Of the outcomes analyzed, cases were significantly more likely to develop non-CP infections within one year of CP than controls (Hazard Ratio = 12.6, 95% confidence interval = 3.1-51.7; P < 0.001). These infections were often bacterial and occurred long after CP infection. In conclusion, CP is uncommon after PLT and has a low likelihood of organ dissemination. No risk factors were identified. Some cases required prolonged hospitalizations. Close monitoring for the development of late bacterial infections is warranted. PMID- 16315313 TI - Liver transplantation for erythropoietic protoporphyria liver disease. AB - In erythropoietic protoporphyria (EPP), there is excessive production of protoporphyrin, primarily in the bone marrow, resulting in increased biliary excretion of this heme precursor. Some patients will develop progressive liver disease that may ultimately require liver transplantation. However, excessive production of protoporphyrin by the bone marrow continues after transplantation, which may cause recurrent disease in the allograft. This study was performed to define post-transplant survival, the risk of recurrent disease, and specific management issues in patients transplanted for EPP liver disease. The patients studied consisted of twelve males and eight females, with an average age of 31 (range, 13-56) years at the time of transplantation. The estimated maximum MELD score prior to transplant was 21 (range, 15-29). Unique complications in the perioperative period were light induced tissue damage in four patients and neuropathy in six, requiring prolonged mechanical ventilation in four. Patient and graft survival rates were 85% at 1 year, 69% at 5 years, and 47% at 10 years. Recurrent EPP liver disease occurred in 11 of 17 patients (65%) who survived more than 2 months. Three patients were retransplanted at 1.8, 12.6, and 14.5 years after the initial transplant for recurrent EPP liver disease. In conclusion, the 5-year patient survival rate in patients transplanted for EPP liver disease is good, but the recurrence of EPP liver disease appears to diminish long term graft and patient survival. PMID- 16315314 TI - Impact of cytomegalovirus prophylaxis on rejection following orthotopic liver transplantation. AB - With improved cytomegalovirus (CMV) prophylaxis, CMV disease after liver transplantation has decreased dramatically, and patient and graft survival have improved. We examined the impact of CMV prophylaxis on biopsy proven rejection after orthotopic liver transplantation by analyzing data on 192 liver recipients over 5 years (1994-1999). Risk factors assessed for biopsy proven rejection including donor and recipient age, CMV serostatus; CMV prophylaxis; immunosuppression; bacteremia and blood product use were examined over a 2-year follow-up. Multivariate analysis of risk factors for rejection showed that bacteremia (HR 3.57, 95% CI 1.39-9.36, P=0.008), donor age (HR 1.20, 95% CI 1.06 1.36, per 10 year increase, P=0.004), and use of cyclosporine as initial immunosuppression compared to tacrolimus (HR 1.98, 95% CI 1.27-3.09, P=0.003) were associated with increased risk; ganciclovir prophylaxis for 3 months (HR 0.51, 95% CI 0.33 to 0.79, P=0.003) and recipient age (HR 0.78; 95% CI 0.63-0.96, for each 10 year increase, P=0.03) were associated with decreased risk. We conclude that, the use of CMV prophylaxis with ganciclovir significantly reduces the incidence of biopsy proven rejection in liver transplant recipients. PMID- 16315315 TI - Regulation of DNA repair gene expression in human cancer cell lines. AB - Although most advanced cancers are incurable, the majority of testicular germ cell tumors can be cured using cisplatin-based combination chemotherapy. The nucleotide excision repair (NER) pathway removes most DNA adducts produced by cisplatin, and the low levels of NER in testis tumor cells may explain why these cancers are curable. Three NER proteins: ERCC1, XPF, and XPA, are present at low levels in testis tumor cell lines, and addition of these proteins to protein extracts of testis tumor cells increases their in vitro DNA repair capacity to normal levels. The aim of this study was to identify the mechanism responsible for the low levels of these DNA repair proteins. The levels of the mRNA transcripts for ERCC1, XPF, and XPA were measured in a panel of 14 different human cancer cell lines, using real-time PCR. Three ERCC1 splice variants were identified and quantitated. Three alternative transcription start points (TSPs) were identified for ERCC1 but none were testis-specific. The significantly lower levels of ERCC1, XPF, and XPA protein in testis tumor cell lines cannot be explained solely by differences in transcriptional efficiency or mRNA stability. For ERCC1, post-transcriptional control by alternative splicing does not account for the testis-specific low levels of protein expression. Pulse-chase experiments showed that the half-life of ERCC1 protein in a testis tumor cell line was not significantly different to that in a prostate cancer cell line. Taken together, these results suggest that constitutive levels of these DNA repair proteins are controlled at the level of translation. PMID- 16315316 TI - Will my protein crystallize? A sequence-based predictor. AB - We propose a machine-learning approach to sequence-based prediction of protein crystallizability in which we exploit subtle differences between proteins whose structures were solved by X-ray analysis [or by both X-ray and nuclear magnetic resonance (NMR) spectroscopy] and those proteins whose structures were solved by NMR spectroscopy alone. Because the NMR technique is usually applied on relatively small proteins, sequence length distributions of the X-ray and NMR datasets were adjusted to avoid predictions biased by protein size. As feature space for classification, we used frequencies of mono-, di-, and tripeptides represented by the original 20-letter amino acid alphabet as well as by several reduced alphabets in which amino acids were grouped by their physicochemical and structural properties. The classification algorithm was constructed as a two layered structure in which the output of primary support vector machine classifiers operating on peptide frequencies was combined by a second-level Naive Bayes classifier. Due to the application of metamethods for cost sensitivity, our method is able to handle real datasets with unbalanced class representation. An overall prediction accuracy of 67% [65% on the positive (crystallizable) and 69% on the negative (noncrystallizable) class] was achieved in a 10-fold cross validation experiment, indicating that the proposed algorithm may be a valuable tool for more efficient target selection in structural genomics. A Web server for protein crystallizability prediction called SECRET is available at http://webclu.bio.wzw.tum.de:8080/secret. PMID- 16315317 TI - Structural specificity in a FGF7-affinity purified heparin octasaccharide required for formation of a complex with FGF7 and FGFR2IIIb. AB - Variations in sulfation of heparan sulfate (HS) affect interaction with FGF, FGFR, and FGF-HS-FGFR signaling complexes. Whether structurally distinct HS motifs are at play is unclear. Here we used stabilized recombinant FGF7 as a bioaffinity matrix to purify size-defined heparin oligosaccharides. We show that only 0.2%-4% of 6 to 14 unit oligosaccharides, respectively, have high affinity for FGF7 based on resistance to salt above 0.6M NaCl. The high affinity fractions exhibit highest specific activity for interaction with FGFR2IIIb and formation of complexes of FGF7-HS-FGFR2IIIb. The majority fractions with moderate (0.30-0.6M NaCl), low (0.14-0.30M NaCl) or no affinity at 0.14M NaCl for FGF7 supported no complex formation. The high affinity octasaccharide mixture exhibited predominantly 7- and 8-sulfated components (7,8-S-OctaF7) and formed FGF7-HS FGFR2IIIb complexes with highest specific activity. Deduced disaccharide analysis indicated that 7,8-S-OctaF7 comprised of DeltaHexA2SGlcN6S in a 2:1 ratio to a trisulfated and a variable unsulfated or monosulfated disaccharide. The inactive octasaccharides with moderate affinity for FGF7 were much more heterogenous and highly sulfated with major components containing 11 or 12 sulfates comprised of predominantly trisulfated disaccharides. This suggests that a rare undersulfated motif in which sulfate groups are specifically distributed has highest affinity for FGF7. The same motif also exhibits structural requirements for high affinity binding to dimers of FGFR2IIIb prior to binding FGF7 to form FGF7-HS-FGFR2IIIb complexes. In contrast, the majority of more highly sulfated HS motifs likely play FGFR-independent roles in stability and control of access of FGF7 to FGFR2IIIb in the tissue matrix. PMID- 16315318 TI - Gene expression in the cell cycle of human T lymphocytes: I. Predicted gene and protein networks. AB - The key genes involved in the cell cycle of human T lymphocytes were identified by iterative searches of gene-related databases, as derived also from DNA microarray experimentation, revealing and predicting interactions between those genes, assigning scores to each of the genes according to numbers of interaction for each gene weighted by significance of each interaction, and finally applying several types of clustering algorithms to genes basing on the assigned scores. All clustering algorithms applied, both hierarchical and K-means, invariably selected the same six "leader" genes involved in controlling the cell cycle of human T lymphocytes. Relations of the six genes to experimental data describing switching between stages of cell cycle of human T lymphocytes are discussed. PMID- 16315319 TI - Gene expression in the cell cycle of human T-lymphocytes: II. Experimental determination by DNASER technology. AB - Human lymphocytes gene expression before and after PHA stimulation is monitored by DNASER technology, a novel bioinstrumentation entirely constructed in our laboratories as previously reported. The validity of the DNASER measurements is confirmed by standard fluorescence microscopy equipped with CCD. The human lymphocytes gene expression here experimentally probed using commercially available DNA microarrays such as Human Starter, appears compatible both with independent bioinformatic prediction and with existing experimental data, pointing to MYC as the key gene in the G0-G1 transition induced by PHA in resting lymphocytes. It does not escape our notice that in cell biology and cancer research DNASER technology based on microarray constructed with few leader genes identified from bioinformatics represents a meaningful cost-effective route alternative to massive frequently misleading molecular genomics. PMID- 16315320 TI - Intracellular proteolytic activity of cathepsin B is associated with capillary like tube formation by endothelial cells in vitro. AB - The lysosomal cysteine protease cathepsin B is implicated in degradation of extracellular matrix (ECM), a crucial step in a variety of physiological and pathological processes, including tumor dissemination and angiogenesis. In this study, we analyzed the contribution of extracellular and intracellular cathepsin B activity on the formation of capillary-like tubular structures by human umbilical vein endothelial cells (HUVECs) grown on Matrigel matrix, using general and specific cysteine protease inhibitors. We demonstrated, by confocal assay using quenched fluorescent protein substrate DQ-collagen IV, that endothelial cells degrade ECM both intracellularly and pericellularly. Intracellular cathepsin B activity detected by degradation of Z-Arg-Arg cresyl violet substrate was co-localized with the products of DQ-collagen IV degradation in the perinuclear region and in the capillary-like tubular structures. Treatment of cells with membrane-permeable CA-074 Me effectively abolished intracellular cathepsin B activity, and resulted in reduced tube length (32.3+/-9.4% at 10 microM), total tubule area (49.6+/-12.4% at 10 microM), and the number of branch points of tubules (47.5+/-7.7% at 10 microM) in a dose-dependent manner. In contrast, CA-074 (0.1-10 microM), a membrane-impermeable cathepsin B specific inhibitor, general cysteine protease inhibitors chicken cystatin (5 microM) and E 64 (10 microM), and the metalloprotease inhibitor Minocycline (10 microM) showed no significant inhibitory effect in our angiogenesis model. These results show that, besides multiple regulatory molecules, intracellular cathepsin B also contributes to the neovascularization process and should be considered as a potential therapeutic target. PMID- 16315321 TI - Mechanical stimuli and nutrients regulate rapamycin-sensitive signaling through distinct mechanisms in skeletal muscle. AB - The mammalian target of rapamycin (mTOR) has been identified as a growth factor and nutrient-sensitive molecule that controls the translational machinery and cell growth. Rapamycin-sensitive (RS) signaling events have also been shown to be necessary for mechanical load-induced growth of skeletal muscle, but the mechanisms involved in the mechanical activation of RS signaling are not known. The finding that mechanical stimuli induce nutrient uptake in skeletal muscle raises the possibility that mechanically induced RS signaling is mediated via a nutrient-dependent mechanism. To investigate this hypothesis, skeletal muscles (ex vivo) were stimulated with nutrients or intermittent mechanical stretch and the phosphorylation of p70S6k [P-p70(389)], PKB [P-PKB], mTOR [P-mTOR(2481)], and p38 [P-p38] was assessed. In comparison to vehicle-treated controls, both nutrient and mechanical stimuli induced P-p70(389), neither stimulus altered P PKB or P-mTOR(2481), and only mechanical stimuli induced P-p38. The nutrient and mechanically induced increase in P-p70(389) was blocked by rapamycin, but only nutrient-induced signaling to P-p70(389) was blocked by wortmannin. Furthermore, the mechanically induced increase in P-p70(389) was not impaired by the removal of exogenous nutrients. Taken together, these results indicate that exogenous nutrients are not required for mechanically induced RS signaling and that nutrient and mechanical stimuli activate RS signaling through distinct upstream mechanisms. PMID- 16315322 TI - Satellite cell numbers in young and older men 24 hours after eccentric exercise. AB - We tested the hypothesis that the expansion of satellite cell numbers, 24 h after maximal eccentric knee extensor exercise, is blunted in older men. Muscle biopsies were obtained from the vastus lateralis of 10 young (23-35 years) and 9 older (60-75 years) men. Satellite cells were identified immunohistochemically using an antibody to neural cell adhesion molecule. After 92 maximal eccentric contractions, the mean number of satellite cells per muscle fiber increased to a greater extent among the young men (141%; P < 0.001) than older men (51%; P = 0.002) from preexercise levels. Similar results were obtained when satellite cells were expressed as a proportion of all sublaminar nuclei. We conclude that a single bout of maximal eccentric exercise increases satellite cell numbers in both age groups, with a significantly greater response among the young men. These data suggest that age-related changes in satellite cell recruitment may contribute to muscle regeneration deficits among the elderly. PMID- 16315323 TI - Pure neural leprosy: diagnostic value of the polymerase chain reaction. AB - Pure neural leprosy (PNL) is often difficult to diagnose when acid-fast bacilli (AFB) cannot be detected. We undertook the present study to evaluate use of the polymerase chain reaction (PCR) in diagnosing PNL. Fifty-eight patients (41 men and 17 women) suspected of pure neural leprosy (PNL) were examined. Patients were classified as borderline tuberculoid (BT, 40 cases) and polar tuberculoid (TT, 18 cases) types. Nerve biopsy was performed and was positive for AFB in 20 patients (all BT patients), i.e., 34.5% of total cases. DNA was extracted from the nerve biopsy samples and amplified using PCR for a specific repeated sequence of DNA from Mycobacterium leprae. PCR analysis was positive in the nerve samples from 29 patients (50%), 27 of the BT type, and 2 of the TT type patients. Further, PCR analysis was positive in 14 of 38 cases that were negative for AFB by nerve biopsy, of which 12 were of the BT type and 2 the TT type. PCR analysis proved to be a useful method to investigate pure neural leprosy, enabling confirmation of the diagnosis in more than a third of the cases that were negative for AFB by nerve biopsy. PMID- 16315324 TI - Assessment of cortical excitability using threshold tracking techniques. AB - Conventional paired-pulse transcranial magnetic stimulation (TMS) techniques of assessing cortical excitability are limited by fluctuations in the motor evoked potential (MEP) amplitude. The aim of the present study was to determine the feasibility of threshold tracking TMS for assessing cortical excitability in a clinical setting and to establish normative data. Studies were undertaken in 26 healthy controls, tracking the MEP response from abductor pollicis brevis. Short interval intracortical inhibition (SICI) occurred up to an interstimulus interval (ISI) of 7-10 ms, with two distinct peaks evident, at ISIs of < or =1 and 3 ms, followed by intracortical facilitation to an ISI of 30 ms. Long-interval intracortical inhibition (LICI) occurred at ISIs of 50-300 ms, peaking at 150 ms. The present study has confirmed the effectiveness of the threshold tracking TMS technique in reliably and reproducibly measuring cortical excitability. Simultaneous assessment of upper and lower motor neuronal function with threshold tracking techniques may help to determine the site of disease onset and patterns of progression in neurodegenerative diseases. PMID- 16315325 TI - Nanoliter scale microbioreactor array for quantitative cell biology. AB - A nanoliter scale microbioreactor array was designed for multiplexed quantitative cell biology. An addressable 8 x 8 array of three nanoliter chambers was demonstrated for observing the serum response of HeLa human cancer cells in 64 parallel cultures. The individual culture unit was designed with a "C" shaped ring that effectively decoupled the central cell growth regions from the outer fluid transport channels. The chamber layout mimics physiological tissue conditions by implementing an outer channel for convective "blood" flow that feeds cells through diffusion into the low shear "interstitial" space. The 2 microm opening at the base of the "C" ring established a differential fluidic resistance up to 3 orders of magnitude greater than the fluid transport channel within a single mold microfluidic device. Three-dimensional (3D) finite element simulation were used to predict fluid transport properties based on chamber dimensions and verified experimentally. The microbioreactor array provided a continuous flow culture environment with a Peclet number (0.02) and shear stress (0.01 Pa) that approximated in vivo tissue conditions without limiting mass transport (10 s nutrient turnover). This microfluidic design overcomes the major problems encountered in multiplexing nanoliter culture environments by enabling uniform cell loading, eliminating shear, and pressure stresses on cultured cells, providing stable control of fluidic addressing, and permitting continuous on-chip optical monitoring. PMID- 16315326 TI - Effect of substrate concentration on dual-species biofilm population densities of Klebsiella oxytoca and Burkholderia cepacia in porous media. AB - The long-term operation of bioremediation technologies relies on the success of the contaminant-degrading microorganism(s) to compete for available resources with microorganisms already present in an aquifer or those that may contaminate a bioreactor. Though research has been performed studying the interaction of multiple species in batch and chemostat reactors, little work has been done looking at multi-species interactions in environments that more closely resemble field-scale applications. The research presented herein examined the interaction of Burkholderia cepacia PR1-pTOM(31c), an aerobic trichloroethylene (TCE) degrading bacterium, with Klebsiella oxytoca, a facultative bacterium, in a flow through porous media (PM) reactor. Growth characteristics and population distributions in PM were compared to previously reported values from batch and chemostat reactors. The faster growing organism in batch experiments (K. oxytoca) did not always have the greater population density in dual-species PM experiments. The biofilm population distribution was influenced by substrate concentration, with B. cepacia having a greater dual-species population density than K. oxytoca at a low (30 mg/L dissolved organic carbon [DOC]) substrate concentration and K. oxytoca having a greater population density at a high (700 mg/L DOC) substrate concentration. This change in species population distribution with change in substrate concentration, which was not observed in batch reactors, was also observed in chemostat reactors. Therefore, manipulation of substrate concentration enabled the control of species dominance to the advantage of the TCE degrading population in this dual-species PM system and may provide a mechanism to enhance bioremediation scenarios involving TCE or other contaminants of concern. PMID- 16315327 TI - Oxidation of aminonitrotoluenes by 2,4-DNT dioxygenase of Burkholderia sp. strain DNT. AB - Aminonitrotoluenes form rapidly from the reduction of dinitrotoluenes (DNTs) which are priority pollutants and animal carcinogens. For example, 4-amino-2 nitrotoluene (4A2NT) and 2A4NT accumulate from the reduction of 2,4-DNT during its aerobic biodegradation. Here, we show that 2,4-DNT dioxygenase (DDO) from Burkholderia sp. strain DNT oxidizes the aminonitrotoluenes 2A3NT, 2A6NT, 4A3NT, and 5A2NT to 2-amino-3-nitrobenzylalcohol, 2-amino-4-nitro-m-cresol and 3-amino-5 nitro-p-cresol, 4-amino-3-nitrobenzylalcohol and aminonitrocresol, and 2-amino-5 nitro-o-cresol, respectively. 2A5NT and 3A4NT are oxidized to aminonitrocresols and/or aminonitrobenzylalcohols, and 4A2NT is oxidized to aminonitrocresol. Only 2A4NT, a reduced compound derived from 2,4-DNT, was not oxidized by DDO or its three variants. The alpha subunit mutation I204Y resulted in two to fourfold faster oxidization of the aminonitrotoluenes. Though these enzymes are dioxygenases, they acted like monooxygenases by adding a single hydroxyl group, which did not result in the release of nitrite. PMID- 16315328 TI - Diels-Alder ligation and surface immobilization of proteins. PMID- 16315329 TI - Complex sequence dependence by excess-electron transfer through DNA with different strength electron acceptors. PMID- 16315330 TI - Hollow and polygonous microtubes of monocrystalline indium germanate. PMID- 16315331 TI - Substrate ambiguity and catalytic promiscuity within a bacterial proteome probed by an easy phenotypic screen for aldehydes. PMID- 16315332 TI - Highly enantioselective addition of diphenylzinc to aliphatic and aromatic aldehydes catalyzed by a readily available H8-binol derivative. PMID- 16315333 TI - Characterization of tumour-infiltrating lymphocytes and apoptosis in colitis associated neoplasia: comparison with sporadic colorectal cancer. AB - The development of colorectal cancer is a major complication for patients with chronic idiopathic colitis. Colitis-associated tumours tend to occur at a younger age and be more aggressive than sporadic colorectal cancers. While we have previously associated the presence of tumour-infiltrating lymphocytes (TILs) and increased apoptosis in sporadic colorectal cancer with high-level microsatellite instability and improved prognosis, little is known of the relationship between these variables in colitis-associated colorectal cancer. The aim of this study was to correlate TILs and tumour cell apoptosis in colitis-associated neoplasms stratified according to microsatellite instability. Twenty tumour and 11 dysplastic samples resected from 21 patients with long-standing colitis were analysed for microsatellite instability at 10 microsatellite markers. TIL distribution (CD3, CD8) and function (granzyme B) were quantified by immunohistochemistry. Neoplastic cell apoptosis was assessed using the M30 CytoDEATH antibody. These findings were compared with 40 microsatellite stable (MSS) sporadic colorectal cancers previously evaluated for TILs and neoplastic apoptosis. Low-level microsatellite instability was found in 1/20 colitis associated tumours. All other colitis-associated lesions were designated MSS. CD3(+) and CD8(+) TIL counts were significantly higher in colitis-associated lesions compared with MSS sporadic colorectal cancer (p < 0.0001, p = 0.001 respectively). Despite their higher TIL density, colitis-associated tumours were more likely to present late (Dukes' stage C or D) (p = 0.02). Functionally, colitis-associated TILs demonstrated significantly less granzyme B expression compared to sporadic cancers (p = 0.002). The level of tumour cell apoptosis was similar between the two groups (sporadic, 1.53%; colitis cancers, 1.45%). In conclusion, MSS colitis-associated tumours have a higher prevalence of CD3(+)/CD8(+) TILs but no associated increase in tumour cell killing by apoptosis. Unlike cytotoxic T cells in sporadic colorectal cancer, TILs do not appear to enhance the prognosis of colitis-associated colorectal cancer. This may be related to an impairment of granzyme B expression within these lesions. PMID- 16315334 TI - What is the current evidence for antigen involvement in the development of chronic lymphocytic leukemia? AB - For many years it has been evident that B-cell chronic lymphocytic leukemia (CLL) displays preferential usage of individual immunoglobulin (Ig) variable heavy chain (V(H)) genes. The V(H)1-69 gene was the first to be reported overrepresented in a large number of CLL patients, where the V(H)1-69(+) CLL rearrangements showed characteristic molecular features, such as unmutated V(H) genes, usage of specific diversity/joining gene segments, and a longer than average complementarity determining region (CDR) 3 with certain common amino acid motifs. Also, biased usage of the V(H)3-07 and V(H)4-34 genes with specific rearrangement characteristics was reported in CLL. These findings led to the speculation that antigens could be involved during CLL development by triggering proliferation of B-cells with specific B-cell receptors (BCRs) leading to an increased risk of transforming events. Recently, we characterized a subset of CLL utilizing the V(H)3-21 gene that also displayed peculiar Ig features, e.g. very short and homologous CDR3s, predominant lambda expression and preferential V(lambda)2-14 gene usage. This V(H)3-21(+) subgroup also had poor prognosis despite the fact that two-thirds of cases carried mutated V(H) genes. Moreover, we and others have thereafter described further CLL subsets with very similar heavy and light chain gene rearrangement features. These latter findings of subsets expressing restricted BCRs have emphasized the hypothesis that antigens could play a role during the pathogenesis of CLL. Interestingly, recombinant antibodies produced from these restricted subsets showed similar cytoplasmatic reactivity within each group, thus suggesting recognition of a limited number of autoantigens. Further characterization of antigens is now necessary in order to understand their nature and exact role in CLL development. PMID- 16315335 TI - Spectral karyotyping, fluorescence in situ hybridization and molecular genetic analysis of de novo partial trisomy 7p (7p15.1 --> pter) and partial monosomy 9p (9p22 --> pter). PMID- 16315336 TI - Concomitant fetal anemia and thrombocytopenia due to anti-D and anti-HPA1a alloimmunization. PMID- 16315337 TI - Treatment with statins after acute myocardial infarction in patients >or=80 years: underuse despite general acceptance of drug therapy for secondary prevention. AB - PURPOSE: It has not been decided to what extent the results from statin trials should be transferred to clinical practice in the very old. The aim of the study was to assess the use of cardiovascular drugs after an acute myocardial infarction (MI), with particular focus on statins, in very old patients as compared to younger patients. METHOD: A sample of 901 acute MI patients was drawn from 16 hospitals in 1999/2000; the patients were followed up for 2.5 years. Information on demographic variables and drug therapy was obtained from hospital records, and in the follow-up period by direct patient contact or questionnaire. The main indications for prescribing the various cardiovascular drugs were recorded. RESULTS: At discharge, drug use in patients >or=80 and <80 years, respectively, was as follows: ACE-inhibitors 48 versus 32%, nitrates 55 versus 32%, diuretics 64 versus 26%, aspirin 72 versus 86%, and beta-blockers 67 versus 85%. A striking difference was found for statins: 9% in the very old and 72% in younger patients. The pattern of drug use generally remained unchanged after 2.5 years. Survival rates for patients >or=80 and <80 years: at discharge 72 versus 90%, after 2.5 years 34 versus 73%. CONCLUSIONS: Drug therapy was widely accepted for the indication secondary prevention after MI in patients above 80 years of age. The various cardiovascular drugs were prescribed to about the same extent for very old and younger patients. The exception was lipid lowering drugs which, despite the physicians' recognition of the indication secondary prevention in the very old patients, were prescribed to a limited extent. PMID- 16315338 TI - Endoscopic sphincterotomy and temporary internal stenting for bile leaks following complex hepatic trauma. AB - BACKGROUND: Biliary leak secondary to blunt or penetrating hepatic trauma and damage to the intrahepatic biliary tree remains a challenging problem. The role and safety of endoscopic retrograde cholangiopancreatography (ERCP) and stenting in this setting were studied. METHODS: All trauma victims who developed a bile leak secondary to hepatic trauma were included. Bile leak was defined as the appearance of bile in a surgical wound or intra-abdominal drain after surgery, following percutaneous drainage of a perihepatic bile collection, or evidence of a leak on hepatobiliary scintigraphy. ERCP was performed within 24 h of diagnosis and included biliary sphincterotomy and internal stenting. Recovery was defined as cessation of leakage. RESULTS: Between 1996 and 2004, six patients with penetrating injuries and five with blunt abdominal injuries were treated according to the study protocol. Eight underwent surgery to control bleeding or for additional intra-abdominal injuries. All bile leaks resolved completely within 10 days of ERCP. One patient died from pulmonary sepsis; ten recovered without hepatobiliary sequelae. CONCLUSION: ERCP, biliary sphincterotomy and temporary internal stenting, together with percutaneous drainage of intra abdominal or intrahepatic bile collections, represent a safe and effective strategy for the management of bile leaks following both blunt and penetrating hepatic trauma. PMID- 16315339 TI - Increased natural killer cell activity in patients with an abdominal aortic aneurysm. AB - BACKGROUND: Natural killer (NK) cells have an emerging role in the development of chronic disease and in the direction and maintenance of inflammatory responses. Abdominal aortic aneurysms (AAA) is a chronic inflammatory disorder of unknown aetiology. The aim was to investigate whether NK cells showed altered function in patients with an AAA. METHODS: The presence, phenotype and function of peripheral blood and tissue NK cells from patients with an AAA, peripheral vascular disease (PVD) and healthy age-sex-matched controls were assessed before and after surgery. RESULTS: Patients with an AAA had significantly higher (P < 0.010) percentages of peripheral blood NK cells (mean (95 per cent c.i.) 23.8 (2.6) per cent) than patients with PVD (17.4 (2.9) per cent) and control subjects (16.2 (2.8) per cent). The NK cells from patients with an AAA had increased cytotoxicity on a per cell basis towards both an NK-sensitive target cell line and human aortic smooth muscle cells. Increased NK cell proportions (22.7 (3.5) per cent) and cytotoxic activity, together with higher C-reactive protein values, persisted after successful AAA repair. CONCLUSION: These data support the hypothesis that increased NK cytotoxicity could be a contributing factor in the generation or potentiation of inflammation in patients with an AAA. PMID- 16315340 TI - Analysing duration of episodes of pharmacological care: an example of antidepressant use in Danish general practice. AB - Analysing duration of treatment episodes has become a standard task in many pharmacoepidemiological studies. However, such analyses are often carried out in a rather simplistic manner and more subtle issues are often ignored. In this paper, methods of analysing duration treatment episodes beyond simple analyses allowing investigation of the risk for certain events over time are demonstrated. In particular, the use of cumulative incidence functions, cause-specific hazard functions, hazard rate models and expected mortality in analysing duration of episodes is presented. We used these statistical techniques in analysing the early treatment history of patients who started a regular treatment with antidepressant drugs in the primary health care sector in Denmark. We have extracted some important features: The risk of discontinuing and switching treatment was very high around 10 weeks after starting treatment. After discontinuing the first treatment period, many patients soon started a second treatment period depending on the duration of the first treatment period with highest risk around 10 weeks. The mortality rate among the patients in treatment was about three times higher than the expected mortality. The risk of dying immediately after stopping treatment was about twice the expected mortality. The analysis suggests that: (1) there is a critical period for a first discontinuing, switching and restarting treatment around 10 weeks, (2) the GPs prescribing habits have more influence on the patterns than patient or drug characteristics, (3) over time Danish GPs tend to prolong the duration of first treatment period and avoid longer treatment breaks. PMID- 16315341 TI - Where East meets West. PMID- 16315342 TI - Balancing act. PMID- 16315343 TI - Lessons in professorship. PMID- 16315344 TI - Current awareness in prenatal diagnosis. PMID- 16315345 TI - The direct and indirect effects of insectivory by birds in two contrasting Neotropical forests. AB - A goal among community ecologists is to predict when and where trophic cascades occur. For example, several studies have shown that forest birds can limit arthropod abundances on trees, but indirect effects of bird predation (i.e. decreased arthropod damage to trees) are not always observed and their context is not well understood. Because productivity is one factor that is expected to influence trophic cascades, we compared the extent to which birds indirectly limit herbivore damage to trees in two lowland Neotropical forests that differed in seasonality of leaf production and rainfall. We compared the effects of bird predation on local arthropod densities and on damage to foliage through a controlled experiment using bird exclosures in the canopy and understory of two forests. We found that birds decreased local arthropod densities and leaf damage in the canopy of the drier site during periods of high leaf production, but not in the wetter forest where leaf production was low and sporadic throughout the year. Birds had no effect on arthropod abundances and leaf damage in the understory where leaf production and turnover rates were low. In support of these experimental interpretations, although we observed that arthropod densities were similar at the two sites, bird densities and the rate at which birds captured arthropods were greater at the drier, seasonally productive site. The influence of top-down predation by birds in limiting herbivorous insects appears to be conditional and most important when the production and turnover of leaves are comparatively high. PMID- 16315346 TI - Testimony before the Subcommittee on Criminal Justice, Drug Policy and Human Resources: Committee on Government Reform United States House of Representatives. PMID- 16315347 TI - Fourth International Workshop on immunology of pre-eclampsia, December 2004, Reunion, France. AB - The Fourth International Workshop on 'Reproductive Immunology, Immunological Tolerance and Immunology of Pre-eclampsia' was held in the Island of La Reunion in December 2004. Besides intense sharing of immunological data, it included also epidemiological studies from South Africa, USA and Australia, as well as genetic studies from South Africa and Mauritius, discussions on the future of graft transplant tolerance, implications of inositol-phosphoglycans in the pathogenesis of pre-eclampsia (PE), pathways to the inflammatory syndrome and other topics. Participants shared the belief that we may be witnessing significant steps forward in our comprehension of PE as an immunological event, with a prominent role for deregulation of the innate immune system, probably controlled by T cells and cytokine networks at the feto-placental interface. There was a growing consensus for an NK cell (KIR)-dependent event, with regulation exerted by T cells, as well as an important role for HLA-C presentation/recognition and HLA-G in mediating inflammatory cytokine imbalance, with an emphasis on IL-12 and IL-16 It is hoped that the next Workshop in 2006 will prove the immune involvement by dissecting the NK/Treg/Ts cell and HLA-CI HLA-G circuits leading to cytokine and vascular dysfunction. PMID- 16315348 TI - Follow-up skeletal surveys prove to be valuable in evaluation of child physical abuse. PMID- 16315349 TI - Utility of follow-up skeletal surveys in suspected child physical abuse evaluations. AB - OBJECTIVE: To evaluate the utility of a follow-up skeletal survey in suspected child physical abuse evaluations. METHODS: In this prospective study, follow-up skeletal surveys were recommended for 74 children who, after an initial skeletal survey and evaluation by the Child Abuse Team, were suspected victims of physical abuse. The number and location of the fractures were recorded for the initial skeletal survey and for the follow-up skeletal survey in each case. RESULTS: Forty-eight of the 74 (65%) children returned for a follow-up skeletal survey. The follow-up skeletal survey yielded additional information in 22 of 48 patients (46%). In three patients (6%) the additional information changed the outcome of cases; child abuse was ruled out in one of these patients and abuse was confirmed in two cases. In three other patients, the follow-up skeletal survey refuted tentative skeletal findings, but did not change the outcome because of other physical findings. CONCLUSION: A follow-up skeletal survey identified additional fractures or clarified tentative findings in children who were suspected victims of physical child abuse. The follow-up skeletal survey should be completed on all patients who have an initial skeletal survey performed for suspected physical child abuse and for whom child abuse is still a concern. PMID- 16315350 TI - Problems of studying extinction risks. PMID- 16315351 TI - Bibliography. Current world literature. Diagnostics and techniques. PMID- 16315352 TI - Interagency collaboration between child protection and mental health services: practices, attitudes and barriers. AB - OBJECTIVE: The aim of this paper is to examine some of the factors that facilitate and hinder interagency collaboration between child protection services and mental health services in cases where there is a parent with a mental illness and there are protection concerns for the child(ren). The paper reports on agency practices, worker attitudes and experiences, and barriers to effective collaboration. METHOD: A self-administered, cross-sectional survey was developed and distributed via direct mail or via line supervisors to workers in statutory child protection services, adult mental health services, child and youth mental health services, and Suspected Child Abuse and Neglect (SCAN) Teams. There were 232 completed questionnaires returned, with an overall response rate of 21%. Thirty-eight percent of respondents were statutory child protection workers, 39% were adult mental health workers, 16% were child and youth mental health workers, and 4% were SCAN Team medical officers (with 3% missing data). RESULTS: Analysis revealed that workers were engaging in a moderate amount of interagency contact, but that they were unhappy with the support provided by their agency. Principle components analysis and multivariate analysis of variance (MANOVA) on items assessing attitudes toward other workers identified four factors, which differed in rates of endorsement: inadequate training, positive regard for child protection workers, positive regard for mental health workers, and mutual mistrust (from highest to lowest level of endorsement). The same procedure identified the relative endorsement of five factors extracted from items about potential barriers: inadequate resources, confidentiality, gaps in interagency processes, unrealistic expectations, and professional knowledge domains and boundaries. CONCLUSIONS: Mental health and child protection professionals believe that collaborative practice is necessary; however, their efforts are hindered by a lack of supportive structures and practices at the organizational level. PMID- 16315353 TI - Large B-cell non-Hodgkin lymphoma. PMID- 16315354 TI - Predicting the use of single versus multiple types of violence towards children in a representative sample of Quebec families. AB - OBJECTIVE: The purpose of this study is to determine which factors best predict parental use of single versus multiple types of violence. METHODOLOGY: The study uses data from a Quebec telephone survey conducted with a representative sample of 2,469 mothers of children aged 0-17 years. The interview covered topics such as mother's attitudes and reports of violence towards children, mother's experiences of childhood violence, level of marital discord, and socioeconomic factors. A discriminant function analysis was performed using individual and family factors to predict membership in five distinct groups: (a) no violence reported; (b) reports of psychological aggression only; (c) reports of minor physical violence only; (d) reports of psychological aggression and minor physical violence; and (e) reports of psychological aggression, minor and severe physical violence. RESULTS: One-way ANOVAs revealed that the groups differed significantly on factors such as child's age and gender, mother's attitudes, mother's experiences of childhood violence, level of marital discord, and number of children in the household. Discriminant function analysis revealed that the groups differed in two ways. The first function maximally separated the report of psychological aggression from minor physical violence only. The second function discriminated between the combined reports of all three types of violence versus the reports of minor physical violence only. Mother's attitudes and experiences of childhood violence were the two factors that best distinguished among groups and best predicted group membership. CONCLUSION: These results offer new insights for the prediction of mother's reports of one type versus multiple types of violence and highlight the importance of distinguishing between types and combinations of family violence in future research. PMID- 16315355 TI - SA health minister urged to stop vitamin-peddling doctor. PMID- 16315356 TI - Uganda is learning from its Global Fund grant suspension. PMID- 16315357 TI - Victimization during childhood and revictimization in dating relationships in adolescent girls. AB - OBJECTIVE: Various experiences with violence during childhood and adolescence (parental violence, exposure to marital violence, sexual abuse within and outside the family, sexual harassment at school, community violence, involvement with violent or victimized peers, and previous dating violence) are examined as potential risk factors for psychological, physical, and sexual revictimization in adolescent girls' dating relationships. METHOD: A group of 917 teenage girls (mean age = 16.3) were recruited in 5 high schools located in low to middle socioeconomic areas. Participants were in the 10th and 11th grades, and each completed a self-administered questionnaire. Analyses were performed on the 622 participants who reported having at least one dating partner in the last 12 months. RESULTS: Prevalence rates for past victimization experiences varied from 13% to 43%. Regarding last-year dating victimization, prevalence rates varied from 25% to 37%, depending on the type of violence sustained. Results suggest that extrafamilial experiences with violence are stronger risk factors for recent dating victimization than intrafamilial experiences, especially being sexually harassed by male peers at school and being involved with violent or victimized peers during the year preceding the survey. However, it is important to differentiate between girls who are repeatedly victims of violence in a single, long-term relationship (repeat dating victimization), and girls who are revictimized by different partners (dating revictimization), the former sustaining more frequent physical and psychological violence than the latter. CONCLUSIONS: Findings underline the importance of early prevention of adolescent dating violence. Prevention programs should especially address extrafamilial experiences with violence as important risk factors for victimization in dating relationships, and teach girls strategies to break up abusive relationships. PMID- 16315358 TI - Multidimensional assessment of resilience in mothers who are child sexual abuse survivors. AB - OBJECTIVE: There has been relatively little attention given to positive adaptation following childhood sexual abuse (CSA), and typically such resilience has been explored primarily in the intrapersonal domain. This study explored questions about later resilience following CSA within a multidimensional framework by assessing resilience across intrapersonal, interpersonal, and intrafamilial domains. METHOD: This community sample consisted of 79 mothers with a history of CSA who had a child living at home with them. Participants completed four outcome measures (Center for Epidemiologic Studies--Depression Scale, Parenting Stress Index [PSI] Health Scale, PSI Parenting Competence Scale, and a measure of marital satisfaction). Risk and protective factors examined in relation to outcome included mother's age, socioeconomic status, severity of the CSA experience, coping strategy (avoidance, seeking social support, and problem solving), child characteristics, and spousal/partner support. RESULTS: Results indicated that when multiple adaptational domains were assessed, mothers showed discrepancies in how adequately they functioned across domains. While severity of the CSA experience was only weakly associated with outcome, use of avoidant coping emerged as a significant risk factor and was strongly and consistently associated with negative outcome across domains. Spousal/partner support was a strong protective factor and buffered the relationship between depressive symptoms and parenting competence. Difficult child characteristics were significantly associated with mothers' perceptions of physical health and parenting competence. CONCLUSIONS: The findings suggest the importance of comprehensive multimethod assessments of resilience and extend the knowledge of factors associated with positive outcome. The results highlight the need for further research exploring current contextual risk and protective factors associated with resilience in each domain. PMID- 16315359 TI - Probing the function of Mycobacterium tuberculosis catalase-peroxidase by site directed mutagenesis. AB - Catalase-peroxidase is a multi-functional heme-dependent enzyme which is well known for its ability to carry out both catalatic and peroxidatic reactions. Catalase-peroxidase from Mycobacterium tuberculosis(mtCP) is of particular interest because this enzyme activates the pro-antitubercular drug isoniazid. It is estimated that 2 billion people are infected with M. tuberculosis, the principal causative agent of tuberculosis, and that 2 million people die from the disease each year. The rise of drug-resistant strains continues to be of critical concern and it is well documented that mutations which reduce activity or inactivate mtCP lead to increased levels of isoniazid resistance in M. tuberculosis. The recent determination of the crystal structure for M. tuberculosis mtCP has aided the understanding of how the enzyme functions and provides a three-dimensional framework for testing hypotheses about the roles of various residues in the active site. Here we report site-directed mutagenesis studies of three conserved residues located near the heme of mtCP, His-108, Trp 107 and Trp-321 including the construction of the double mutant W107F-W321F. Resulting mutants have been purified and their catalatic and peroxidatic activities have been determined. Data are compared in the context of related studies aimed at dissecting the roles of these residues in the different activities of the enzyme. Analyses of single and double mutants studied here emphasise that the hydrogen bonding network surrounding the heme in the active site appears more important for maintenance of catalatic rather than peroxidatic activity in CP enzymes. PMID- 16315360 TI - Action through collaboration: a conversation with David Brailer. Interview by Robert Cunningham. AB - Progress toward widespread adoption of health information technology (HIT) by providers remains uncertain and fraught with obstacles. But since his appointment as national coordinator for HIT in May 2004, David Brailer has observed a flowering of initiatives at the state level and the stirring of grassroots demand for action by providers. Brailer sees the creation of a new leadership group to spur adoption of technical standards as an important step forward. He remains concerned that small provider organizations will fall behind larger ones and that IT adoption will move forward without adequate standardization, compromising opportunities for interoperability in the future. PMID- 16315361 TI - Using honey to heal a chronic wound in a patient with epidermolysis bullosa. AB - This case study details the healing of a chronic wound (20 years' duration) in a patient with dystrophic epidermolysis bullosa (EB). Many different dressings and creams had been used, and on occasions the wound began to heal but never progressed to closure. A honey impregnated dressing was used and the wound healed in 15 weeks. A brief overview of the dystrophic form of EB is given and some evidence for the efficacy of honey is presented. PMID- 16315362 TI - CA: nurses may have been negligent in delay: nurses' negligence not cause of patient's death. Arguelles v. Seton Medical Center, 2005 WL 2375628 -CA. PMID- 16315363 TI - Counting the years. PMID- 16315364 TI - Outside the box. This condition frequently goes unrecognised, despite its impact on patients and public health. PMID- 16315365 TI - Welfare writes. Mr Blunkett, you can keep your wellies--I won't be wading into muddy water. PMID- 16315366 TI - Uncertainty and precaution in European environmental risk assessment of chemicals. AB - It is recognised that there is a need for a proper treatment and transparency of uncertainty in risk assessment and management, especially in view of the upcoming proposed new chemical policy REACH, which delegates the responsibility for conducting risk assessments to industry. The current EU risk assessment for new and existing substances is largely deterministic and prudential measures are implicitly embedded in calculation schemes and rules. In this paper, a more probabilistic approach to risk assessment is advocated. The advantage is twofold: 1) inherent variability and other uncertainty pertaining to exposure and effects are transparently taken into account, while at the same time 2) issues of caution are explicitly transferred to the risk management phase. The result of a probabilistic risk assessment as suggested is improved transparency with quantitative and qualitative uncertainty estimates. Such uncertainty information can be used to discuss precautionary measures in the context of risk management. PMID- 16315367 TI - Sporadic inclusion-body myositis: a proposed key pathogenetic role of the abnormalities of the ubiquitin-proteasome system, and protein misfolding and aggregation. AB - Sporadic inclusion-body myositis (s-IBM), the most common muscle disease of older persons, is of unknown cause and there is no successful treatment. We summarize our most recent findings in s-IBM muscle fibers, which demonstrate abnormalities of the ubiquitin-proteasome system, and abnormal accumulation, misfolding and aggregation of proteins. We propose that these changes, possibly provoked by the aging intra-muscle fiber cellular milieu, and aggravated by the oxidative stress, play a key pathogenic role in s-IBM. This evidence strongly suggests that mechanisms other than the immune/inflammatory response play the important role in s-IBM muscle fiber degeneration. PMID- 16315368 TI - Contamination of nebulizers with environmental allergens. AB - BACKGROUND: A previous article described cockroach allergen in the nebulizer reservoir of an asthmatic patient who experienced a life-threatening exacerbation after nebulizer use. OBJECTIVE: To determine whether indoor allergens can be measured in home nebulizers. METHODS: As part of a large study examining nebulizer use in underserved asthmatic children, visiting nurses replaced nebulizer sets in patients' homes. Twenty used sets were randomly selected for analysis, without linkage to clinical or home environmental data. Nebulizer reservoirs and negative controls (buffer and albuterol) were extracted overnight with 2 mL of buffer. For positive controls, nebulizer sets were placed in homes with cats and dogs, and other reservoirs were intentionally contaminated with cat (Fel d 1), dog (Can f 1), cockroach (Bla g 1 and Bla g 2), and mouse (Mus m 1) skin test solutions. Extracts were tested for allergens in a masked manner using enzyme-linked immunosorbent assay. RESULTS: Of 17 reservoirs with adequate specimens for allergen detection, 5 (29%) had measurable levels for at least 1 of 5 allergens tested. One reservoir had measurable Can f 1, 2 had Bla g, 3 had Mus m 1, and none had Fel d 1 allergen. Two of 3 homes with cats where nebulizer setups were placed had measurable Fel d 1 in the reservoir, and 1 of 2 homes with dogs had measurable Can f 1. Reservoirs kept in sealed plastic bags had no detectable allergen. CONCLUSIONS: Indoor allergens can be found in the nebulizer equipment of children with asthma, with the potential for adverse consequences. Storing nebulizer sets in sealed plastic bags may prevent contamination. PMID- 16315369 TI - Predictors of hypertension and changes of blood pressure in HIV-infected patients. AB - OBJECTIVE: We assessed predictors of changes in systolic (SBP) and diastolic (DBP) blood pressure during follow-up and of the development of hypertension in HIV-infected individuals. METHODS: International cohort collaborative study (D:A:D) of established prospective cohorts of HIV-1-infected patients. Longitudinal analysis of changes in blood pressure (BP) was performed using mixed effects models in 17170 patients. Predictors of development of hypertension during follow-up (systolic BP > or =140 and/or diastolic BP > or =90 mmHg or initiation of antihypertensive treatment) were assessed using Cox models in 8 984 patients with a normal BP level at baseline. RESULTS: 73548 BP measurements with a median of 4 per patient (interquartile range [IQR]: 2-6) were recorded over a median follow-up of 2.3 years (IQR: 1.5-2.6). Risk factors significantly associated with a development of higher systolic BP and diastolic BP (differences > or =5 mmHg and P-values <0.001) during follow-up were: older age, male sex, higher body mass index (BMI) and use of BP-lowering drugs. In patients with normal BP at baseline, 1186 developed hypertension for an incidence of 72.1 per 1000 person-years (95% confidence interval: 68.2-76.0). Factors associated with development of hypertension were: male sex, higher BMI, older age, higher BP at baseline, high total cholesterol and clinical lipodystrophy. Cumulative duration of exposure to nucleoside reverse transcriptase inhibitors (P=0.75), protease inhibitors (P=0.92) as well as type of antiretroviral treatment at baseline (P=0.18) were not associated with a higher risk of hypertension. Cumulative duration of exposure to non-nucleoside reverse transcriptase inhibitors (NNRTIs) was significantly associated with lower risk of hypertension (hazard ratio=0.78 and 0.67 for those treated < or =10 months and >10 months compared with no exposure; P=0.005). CONCLUSIONS: Increased blood pressure in HIV-infected individuals is associated with established risk factors for hypertension. There was no evidence for an independent deleterious effect of any class of antiretroviral drugs on BP, although the use of NNRTIs was associated with a lower risk of development of hypertension. PMID- 16315370 TI - Bacterial genomic DNA isolation using sonication for microarray analysis. PMID- 16315371 TI - CpG analyzer, a Windows-based utility program for investigation of DNA methylation. PMID- 16315372 TI - Two-step cross-linking method for identification of NF-kappaB gene network by chromatin immunoprecipitation. AB - The chromatin immunoprecipitation (ChIP) assay has recently been exploited as a powerful and versatile technique for probing protein-DNA interactions within the chromatin environment. In this method, intact cells are fixed with a reversible DNA-protein cross-linking agent (formaldehyde), and associated DNA is enriched by immunoprecipitating a target DNA binding protein. The bound DNA in the immune complexes is then used to identify that specific DNA binding protein's endogenous genomic targets. Nuclear factor kappaB (NF-kappaB) is a highly inducible transcription factor that controls genetic networks important for pathogen- or cytokine-induced inflammation, immune response, and cellular survival. In our studies of the genetic network under control of the inducible NF-kappaB transcription factor, we found that the conventional ChIP technique using a single formaldehyde cross-linking step did not reproducibly cross-link it to DNA. As a result, we have developed a novel ChIP assay using a two-step cross-linking procedure, incorporating N-hydroxysuccinimide (NHS)-ester-mediated protein protein cross-linking prior to conventional DNA-protein cross-linking. We demonstrate that this technique is highly efficient, cross-linking virtually all NF-kappaB/Rel A into covalent complexes, resulting in quantitative and robust identification of inducible NF-kappaB family binding to a variety of validated NF kappaB-dependent genomic targets. To demonstrate the general utility of this two step cross-linking procedure, we performed enhanced capture of cytokine-inducible signal transducer and activator of transcription-3 (STAT3) binding to one of its known target genes. Our method represents a significant improvement in the efficiency of ChIP analysis in the study of endogenous targets for rare transcription factors. PMID- 16315373 TI - In vitro and in vivo antigen expression of Candida albicans. PMID- 16315374 TI - Evaluation of antitumor and antimetastatic activity of pepstatin A in some experimental tumor models. PMID- 16315375 TI - [Changes in distributive equity of health insurance contribution burden]. AB - OBJECTIVES: We analyzed the changes from 1996 to 2002 in distributive equity of the contribution burden in the Korean National Health Insurance. METHODS: The study subjects were a total of 8923 employee households and a total of 7296 self employed households over the period from 1996 to 2002. Those were the households meeting the two criteria as completing each annual survey and having no change in the job of head of the household during that period from the raw data of the Household Income and Expenditure Survey annually conducted by the Korean National Statistical Office. The unit of analysis was a household, and this was the standard for assessing the contribution that is now applied on a monthly basis. Deciles Distribution Ratio, Contribution Concentration Curve and Contribution Concentration Index were estimated as the index of inequality. Multiple regression analysis was conducted to compare the annual ability-to-pay elasticity of the contribution to the reference year of 1996 for three groups (all households, the employee households, and the self-employed households). RESULTS: For the index of inequality, the distributive equity of contribution was improved in all three groups. In particular, the employee group experienced a substantial improvement. Using multiple regression analysis, the ability-to-pay elasticity of the contribution in the employee group significantly increased (beta = 0.232, p < 0.0001) in the year 2002 as compared to the reference year of 1996. The elasticity in the self-employed group also significantly increased (beta = 0.186, p < 0.05), although its change was smaller than that in the employee group. CONCLUSIONS: The employee group had a greater improvement for the distributive equity of the contribution burden than the self-employed group. Within the observation period, there were two important integration reforms: one was the integration of 227 self-employed societies in 1998 and the other was the integration of 139 employee societies in 2000. We expected that the equity of the contribution burden would be improved for the self-employed group since the integration reform of 1998. However, it was not improved for the self-employed group until the year 2000. This result suggests that capturing exactly the beneficiaries' ability-to-pay such as income is the precedent for distributive equity of the contribution burden, although a more sophisticated imposition standard of contribution is needed. PMID- 16315376 TI - Attenuation of exaggerated exercise blood pressure response in African-American women by regular aerobic physical activity. AB - INTRODUCTION: A hyperreactive blood pressure response to exercise is a predictor of developing hypertension. The present study determined the influence of physical activity on an exaggerated exercise blood pressure response (EEBPR) in normotensive African-American women. METHODS: We screened 36 women 18-26 years of age for EEBPR defined as a > or = 50 mm Hg difference in systolic blood pressure at rest and during exercise at 50% peak oxygen uptake (VO2peak). Seven subjects demonstrated an EEBPR and participated in the study. Study participants trained for eight weeks on a bicycle ergometer at a work intensity of 70% VO2peak. Blood pressure, heart rate, cardiac output (CO), stroke volume (SV), and total peripheral vascular resistance (TPR) were determined at baseline and during submaximal exercise at power outputs of 30 W and 50% VO2peak. Subjects served as their own controls, and data were evaluated by using a paired t test at P<.05. RESULTS: Effectiveness of the intervention was shown by a significantly greater VO2peak associated with significant decrements in systolic and mean arterial pressures at power outputs of 30 W and 50% VO2peak. A significant decrement in heart rate was observed during exercise at 30 W. Significant increments in CO and SV and decrement in TPR were found during exercise at 50% VO2peak. CONCLUSION: The reduction in TPR associated with regular aerobic physical activity may attenuate the EEBPR and decrease the risk for hypertension in normotensive, young adult, African-American women. PMID- 16315377 TI - The relationship of racism to appraisals and coping in a community sample. AB - Ethnicity-related maltreatment (racism) is a significant stressor for many Americans and may contribute to racial disparities in health. Mechanisms linking this stressor to health status are not yet understood. This study tests the hypothesis that lifetime exposure to racism influences individuals' appraisals of and coping responses to new episodes of maltreatment. Participants included 420 Black and Latino patients and staff of community primary care practices in New York City. Participants completed the Brief Perceived Ethnic Discrimination Questionnaire--Community Version. They also completed measures of appraisals and anger coping modified to inquire about responses to new episodes of ethnicity related maltreatment. Individuals who had higher levels of lifetime exposure to discrimination were more likely to experience new episodes as threatening and potentially harmful. Exposure to ethnic discrimination was also positively related to the use of anger coping styles, but the magnitude of the relationship varied depending on the type of discrimination. Individuals who had been exposed to higher levels of workplace discrimination were more likely to suppress anger in new situations. Those who were exposed to ethnicity-related social exclusion or harassment were more likely to confront others and aggressively express their feelings. The significance of the relationship held even when controlling for mood and personality variables that might account for both racism and coping. No differences were found between Blacks and Latinos in the relationship of racism to appraisals and coping. These findings add to the growing empirical literature on strategies for coping with racism. PMID- 16315378 TI - A comparison of the sociodemographic, risk-behavior, and substance-abuse profile of young vs older HIV-infected Puerto Rican patients. AB - INTRODUCTION: As of May 2003, 17% of all reported AIDS cases in Puerto Rico had occurred among those 20-29 years of age. These individuals were likely initially infected with HIV in adolescence. The objectives of this study are to describe and compare the prevalence of the sociodemographic, risk-behavior, and substance use profile among patients infected with HIV during their adolescence (early) and compare them with patients infected at an older age (non-early). METHODS: This is a cross-sectional study of 3151 HIV/AIDS patients admitted to the retrovirus research center of our institution between 1992 and 2002. The variables we studied include the presence or absence of early infection, sociodemographic variables, risk-behavior variables, and substance-use variables. An early infected patient was defined as a patient with HIV/AIDS that reported his or her first positive HIV test result before the age of 21 years. Descriptive and differential analyses were performed. RESULTS: Five percent of our patients were early-infected (157/3151). A significantly higher proportion (P < or = .05) among the early-infected patients as compared to the older group was females, required inpatient hospital care at the time of study entry, and had less than a 12th grade education. In the interview a significantly higher proportion had antisocial behavior, had been in prison at some point in their life, and had used crack-cocaine. CONCLUSIONS: The early HIV-infected patients showed a different sociodemographic, risk-behavior, and substance-use profile. Knowledge of the specific characteristics of early HIV-infected patients could be used to develop primary prevention programs directed toward reducing HIV infection among young Puerto Ricans. PMID- 16315379 TI - Racial differences in institutionalization after hip fractures: California hospital discharge data. AB - OBJECTIVE: Hip fractures related to osteoporosis are a major cause of illness leading to institutionalization of elderly patients. Our objective was to determine the ethnic differences and the factors influencing the placement of hip fracture patients in extended-care facilities (ECFs). DESIGN: Secondary data analyses of California Hospital Discharge Data Program (CHDDP). PARTICIPANTS: 78,576 men and 246,177 women > or = 65 years of age with hip fractures related to osteoporosis were included for the analyses. METHODS: The main outcome variable was placement in ECFs as opposed to home. Other variables studied were age, gender, ethnicity, cormobid conditions, insurance, and length of stay in the hospital. RESULTS: Whites and African-American patients with hip fractures were more likely to be placed in ECFs compared to Hispanics. Older age, male gender, presence of two or more comorbid conditions, Medicare insurance, and shorter length of stay in the hospital were also significantly associated with placement in ECFs. CONCLUSION: Future studies should explore sociocultural and other factors that influence ethnic differences in the disposition of hip fracture patients. PMID- 16315380 TI - College-age, African-American males' misperceptions about weight status, body size, and shape. AB - This study examined actual and perceived weight status and body satisfaction of college-age African-American males. We predicted that, in general, males would be accurate about their weight status and satisfied with their body sizes and shapes. The study population was 123 male from a cross-sectional survey of 406 seniors who were scheduled to graduate in the spring 2003 from a historically Black university located in the mid-Atlantic region. These students were administered a survey on health and wellness that addressed a myriad of health concerns, most of which addressed weight, exercise, and related areas. This paper presents analyses on males who were overweight or obese according to the National Institutes of Health guidelines (body mass index > or = 25 kg/m2). 50.4% were overweight or obese (OW/O). Of the OW/O males, 59.7% inaccurately classified their own weight status as normal and chose ideal weights (F(2,59)=3.8, P<.04) and healthy weights (F(2,59)=8.0, P<.001) that were heavier than males with accurate weight perceptions. Specifically, OW/O males desired larger upper torsos (chi2=7.2, df=1, P<.01) and larger body parts (ie, arms, legs, chest area; F(2,59)=11.0, P<.0001). Inaccurate, overweight males were less likely to agree that losing weight supported healthiness (chi2=26.5, df=4, P<.001) or that losing weight would make them more attractive (chi2=14.4, df=4, P<.01). These findings point to the need for effective interventions and strategies for helping those affected and those interested in overweight prevention to understand the role of weight perception, body satisfaction, and their influence on weight loss and lifestyle changes for health. PMID- 16315381 TI - Oral health disparities and periodontal disease in Asian and Pacific Island populations. AB - INTRODUCTION: While oral health disparities exist in many ethnic groups in Hawaii, the challenge of developing research and intervention programs is hampered by the lack of a dental school and adequate state resources. OBJECTIVE: To use a collaboration model to establish a mentoring relationship with a research-intensive school of dentistry to reduce oral health disparities in Hawaii. METHODS: Collaborative interactions with the University of Hawaii School of Medicine (UH) and the University of North Carolina School of Dentistry at Chapel Hill (UNC) included bimonthly teleconferences, on-site planning and mentoring sessions, yearly conferences in Hawaii open to the community using UNC faculty, and on-site skills training sessions. The community was asked to participate in determining priorities for research through focus-group interactions. Two pilot investigations were also conducted. RESULTS: Both universities have been awarded grants to fund activities to support the combined intellectual and physical resources of multiple private, public, and community organizations to achieve the goal of improving the oral health status of the people of Hawaii. As a result of initial planning, two related grants have been submitted (one approved, one disapproved) to fund pilot studies on the oral health status of mothers and their babies in a rural community. These studies include both UH and UNC investigators. CONCLUSIONS: Health disparities occur among diverse ethnic groups in Hawaii, and links between general health and oral health continue to emerge. In spite of obstacles to designing effective research and intervention programs in Hawaii, UH fostered a collaborative relationship with a premiere dental research institution to develop competence in clinical research, conduct pilot studies, and obtain extramural funding for comprehensive studies. Direct involvement of community representatives in the research process is integral to the success of such studies and will continue to serve as the foundation of our community-based participatory research. The network partners have accomplished their primary goal of developing culturally appropriate methods for assessing determinants of oral health, oral health-related quality of life, and health outcomes in Asians and Pacific Islanders. PMID- 16315382 TI - Posttraumatic stress disorder symptoms and adherence among women living with HIV. AB - OBJECTIVES: Posttraumatic stress disorder (PTSD) is an anxiety disorder associated with serious traumatic events. People with PTSD tend to perceive medical interventions as intrusive. Factors such as dose complexity, number of pills, food requirements, patient-health provider relationship, and individual factors influence adherence. This study intends to examine if traumatic experiences affect the way women living with HIV adhere to their medical treatment. METHODS: Adherence to HIV medications was examined with a self-report scale that was administered to a group of women that attend the Maternal Infant Studies Center. Trauma symptoms were measured by using the Trauma Symptom Inventory. RESULTS: A total of 15 women were studied. Of these, 86.6% were found adherent and 13.4% nonadherent. Both the adherent and nonadherent activated the anger/irritability (7.7% vs 50%), sexual concerns (7.7% vs 50%), and tension reduction clinical (TRB) scales (15.4% vs 100%). The adherent women also activated the anxious arousal, depression, defensive avoidance, and dissociation scales. CONCLUSIONS: This study reveals differences in trauma-related symptoms among adherent and nonadherent patients. Increases in the TRB index of 100% among the nonadherent may reflect a need for psychological treatment. By lowering this index, they might improve their desire to live and adhere better to their treatment. Because of the limited sample size, we cannot generalize these findings. We will continue to collect data on a desired sample size of 85 women. PMID- 16315383 TI - Identifying nonrandom occurrences of simple sequence repeats in genomic DNA sequences. AB - Numerous disorders, including prostate cancer and muscular dystrophy, have been associated with nonrandom occurrences of certain simple sequence repeats (SSRs) found in genomic DNA. In a previous paper, we introduced probabilistic methods for identifying such SSRs that possess nonrandom distribution profiles. Here, we apply these methods to the distribution profiles of SSRs of monomers, dimers, trimers, and tetramers occurring in the human genomic sequence data. In particular, we show that the nonrandomness of SSRs is an exponential function of SSR length. We also demonstrate the existence of threshold SSR lengths for the significant nonrandomness (specifically, under/over-representation) of SSRs. These results are consistent with previous findings and demonstrate the potential use of the previously derived probabilistic methods in the identification of (putative disease associated) SSRs that exhibit significant deviations from random expectations. PMID- 16315384 TI - An overview of obesity and diabetes in the diverse populations of the Pacific. AB - BACKGROUND: Pacific islanders (PI) have some of the highest prevalence of obesity and type 2 diabetes in the world. Data related to PI are often reported as though the group is an aggregate; however, PI are a group of very diverse peoples. OBJECTIVE: To explore the obesity problem in PI by: 1) characterizing the major ethnic groups in the Pacific; 2) describing the most recent US census data on PI and their recent immigration patterns; and 3) synthesizing the literature on obesity in PI and type 2 diabetes mellitus. METHODS: Census information and anthropologic resources on the Pacific Islands and its peoples were obtained through an Internet search. Literature on obesity, diabetes, and body composition in PI were obtained through a systematic analysis of citations listed on Medline. RESULTS: Pacific Islanders (PI) are diverse people, culturally, biologically, and linguistically. Polynesians and some Micronesians have a very high prevalence of obesity and diabetes. Melanesians are among the most diverse ethnic groups in the world. Body mass index (BMI) cut-point values for Polynesians may be inaccurate. CONCLUSIONS AND RECOMMENDATIONS: The cause of obesity and diabetes in PI is unclear. Some factors involved are related to urbanization and migration. Obesity research should distinguish between island ethnicities and not aggregate all PI together. Research is needed to define body fat distribution of Polynesians in relationship to obesity co-morbidities, to determine normal growth of Polynesian children, to determine the relationship between growth abnormalities and adult obesity, and to determine the impact of immigration on PI. PMID- 16315385 TI - The hemotoxicity of para-substituted aniline analogs in dog and rat erythrocytes: a species comparison. AB - INTRODUCTION: Hemolytic anemia, the early removal of erythrocytes from the circulation, has been recognized as a side effect of drugs and other environmental chemicals. Formation of methemoglobin (MetHb) following chemical exposure is the first hemotoxic response in the induction of hemolytic anemia. The purpose of this study was to compare the species differences in the chemical induction of MetHb in dog and rat erythrocytes exposed to para-substituted halogenated aniline analogs (phenylhydroxylamine, p-bromo-, p-fluoro-, and p-iodo phenyhydroxylamine). METHODS: Whole blood was collected from a healthy, male Dalmatian dog that weighed 51 lbs and male Sprague-Dawley rats that weighed 100 125 g. Cells were washed (x3) with phosphate-buffered saline supplemented with glucose (pH 7.4). Methemoglobin (MetHb) induction was determined by treating aliquots with pre-specified micromoles of the test agents. Aliquots (75 microL) were removed from each treatment group at specific time points and mixed with cold hemolysis buffer for MetHb determination. Methemoglobin (MetHb) was determined spectrophotometrically at 635 nm. RESULTS: Methemoglobin (MetHb) levels in dog erythrocytes treated with the four analogs increased continuously over 180 minutes and showed no signs of declining. Methemoglobin (MetHb) levels in rat erythrocytes, however, immediately increased and continued to rise and fall before gradually approaching control levels. CONCLUSIONS: Our data demonstrated the species differences in the MetHb-inducing ability of the analogs tested in both dog and rat erythrocytes. The differences in the patterns associated with MetHb induction in the animal models used may be attributed to variations in the MetHb reductase enzyme in both species. PMID- 16315386 TI - Expression of IQ-motif genes in human cells and ASPM domain structure. AB - Genes encoding multiple IQ-motif proteins have been identified in the human genome and may be regulated by calmodulin (CaM). Three genes of unknown function, abnormal spindle-like primary microcephaly (ASPM), KIAA0036, and KIAA1023, were expressed strongly in nearly all transformed human cell lines and in a panel of 16 adult human tissues by reverse transcription polymerase chain reaction. However, ASPM gene expression was not detected in adult brain or skeletal muscle. To better understand function, the domain structure of ASPM was examined. Abnormal spindle-like primary (ASP) protein (abnormal spindle) of Drosophila spp, an orthologue of ASPM, is involved in mitosis, and mutations lead to abnormal spindles and inhibition of cytokinesis. Studies of ASP have indicated that a microtubule binding region exists on the N-terminal third of the protein. Reiterative searches of the protein database using PSI-BLAST identified a common putative microtubular binding domain of 240 residues designated as MTASP. This nearly "all alpha" domain occurs in >25 related proteins including ASP and ASPM. The major C-terminal region of MTASP is basic with conserved hydrophobic residues and terminates at a flanking actin binding (CH) domain. This region is somewhat similar to other microtubule binding proteins such as MAP1B, MAP2, and tau. Multiple IQ motifs and often a conserved C-terminal domain occur in the remaining sequence. The multidomain structure of ASPM suggests a role in the coordination of cell cycle events. The extensive expression of multiple IQ-motif genes and the absence of ASPM in nondividing adult brain and skeletal muscle also suggest a role in cell division. PMID- 16315387 TI - The prevalence of hypovitaminosis D among US adults: data from the NHANES III. AB - OBJECTIVE: Several epidemiologic and mechanistic studies suggest that 25(OH) D3 levels should be maintained above 70 nmol/L for a positive effect on the health of adults. Prior studies have noted low 25(OH) D3 levels in subsets of minority populations. The objective of this study is to examine the prevalence of adequate 25(OH) D3 levels among US adults. METHOD: Using data from the third National Health and Nutrition Examination Survey (NHANES III), we evaluated serum levels of 25(OH) D3 (nmol/L) among 15,390 adult participants > or = 18 years of age. Racial/ethnic grouping was by self-identification as White, Black or African American, and Hispanic. RESULTS: The mean levels of 25(OH) D3 were lower among the female than male participants (71.1 vs 78.7; P=.003) and among the elderly (> or = 65 years of age vs 40-59 and 18-39) than young participants. White men and women (83.0 and 76.0) had higher mean levels of vitamin D than Hispanic men and women (68.3 and 56.7; P<.0001) and than Black men and women (52.2 and 45.3; P<.0001), respectively. The prevalence of both mild-moderate and severe deficiency of vitamin D is higher among women (P<.0001) and minority populations (P<.0001). However, even among White men, 34% had low vitamin D levels. CONCLUSION: Serum levels of 25(OH) D3 are below the recommended levels for a large portion of the general adult population and in most minorities. Need exists for a critical review and probable revision of current recommendations for adult vitamin D intake to maintain adequate 25(OH) D3 levels. PMID- 16315388 TI - The impact of routine vitamin supplementation on serum levels of 25 (OH) D3 among the general adult population and patients with chronic kidney disease. AB - BACKGROUND: Vitamin D supplementation is recommended to maintain bone health in the general population and in particular in patients with chronic kidney disease (CKD). While the nutritional status of vitamin D is assessed by circulating levels of 25 (OH) D3, it is not routinely measured to ensure the adequacy of vitamin D supplementation. Current recommendations require the maintenance of serum levels of 25 (OH) D3 > or = 70 nmol/L. The objective of this study is to assess the effect of routine vitamin supplements on the serum levels of 25 (OH) D3 in the general population and among persons with CKD. METHOD: Using data from the third National Health and Nutrition Examination Survey (NHANES III) we assessed the adequacy of routine vitamin supplementation by assessing serum levels of 25 (OH) D3 among 15,390 adult participants, both with and without CKD. RESULTS: In the general population the participants with vitamin supplements had higher serum level of 25 (OH) D3 (79.47 vs 74.38 nmol/L) and a lower prevalence of vitamin D deficiency (39% vs 48%) than participants not taking any supplements. In the CKD subgroup, the prevalence of vitamin D deficiency was lower with supplements (49%), while greater without supplements (59%). Vitamin D deficiency was higher among women, elderly, and minorities as previously reported. In an adjusted regression model the odds of severe vitamin D deficiency (<25 nmol/L) was 1.43 (P=.0032) among CKD patients, with a trend toward higher rates among patients not taking vitamin supplements (odds ratio 1.47, P=.0557). CONCLUSION: Vitamin supplementation is associated with a lower prevalence of vitamin D deficiency and higher serum levels of 25 (OH) D3. However, the current dose of vitamin D in routine vitamin supplements is still insufficient to maintain adequate serum 25 (OH) D3 levels in a substantial portion of both the general and CKD populations. We must re-asses the dose of vitamin D in routine vitamin supplements in the United States. PMID- 16315389 TI - The research subject advocate at minority Clinical Research Centers: an added resource for protection of human subjects. AB - In early 2001, the National Institutes of Health (NIH) created the research subject advocate (RSA) position as an additional resource for human subjects protection at NIH-funded Clinical Research Centers (CRCs) to enhance the protection of human participants in clinical research studies. We describe the RSA position in the context of clinical research, with a particular emphasis on the role of the RSA in two of the five CRCs funded by the NIH Research Centers in Minority Institutions (RCMI) program. Through participation in protocol development, informed consent procedures, study implementation and follow-up with adverse events, the RSA works closely with research investigators and their staff to protect study participants. The RSA also conducts workshops, training and education sessions, and consultation with investigators to foster enhanced communication and adherence to ethical standards and safety regulations. Although we cannot yet provide substantive evidence of positive outcomes, this article illuminates the value of the RSA position in ensuring that safety of research participants is accorded the highest priority at CRCs. On the basis of initial results, we conclude that the RSA is an effective mechanism for achieving the NIH goal of maintaining the utmost scrutiny of protocols involving human subjects. PMID- 16315390 TI - Morehouse Community Physicians Network (CPN) hypertension registry: patterns of care and opportunities for targeted medical education. AB - BACKGROUND: Provider-focused strategies for improving outcomes in hypertension have produced mixed results. Studies suggest that the effectiveness of a chosen strategy increases when it is tailored to the specific situation. The hypertension registry includes data on African-American hypertensives who receive care in community-based primary care settings. We examined the registry to identify patterns of care and opportunities for provider-focused interventions to improve patient outcomes. METHODS AND RESULTS: The registry will include all records of hypertensive patients from 50 community-based primary care practices at full enrollment. Data from nine practices were manually abstracted into an electronic database and analyzed. Seven hundred and ten records were included in this report. Approximately 70% are female, average age 47 +/- 13 years, 5.3% are uninsured, and more than 60% have at least a high school education. Registry patients have multiple co-morbid conditions: 28% are diabetic, 8% have left ventricular hypertrophy, 5% have congestive heart failure, 6.5% have renal insufficiency, 5% have cerebrovascular disease, 3.5% have previous myocardial infarction and 2% have peripheral vascular disease. Among those with diabetes, mean glycosylated hemoglobin was 7.4 +/- 2. Pattern of antihypertensive use showed 43% on diuretics, 28% on calcium channel blockers, 24% on angiotensin converting enzyme inhibitors, 20% on beta blockers and 16% on angiotensin receptor blockers. Overall, 37% were at goal blood pressure and among those with diabetes, only 16% reached goal blood pressure. CONCLUSION: We conclude that the blood pressure control rates of African Americans in the registry trail those of the general population. This provides a unique opportunity to study the underlying factors and design tailored interventions to address this disparity in health outcome. PMID- 16315391 TI - The Community Physicians' Network (CPN): an academic-community partnership to eliminate healthcare disparities. AB - INTRODUCTION: Disparities in health care are maintained by three primary factors: 1) patient factors which include multiple risk factors and comorbidities; 2) healthcare practitioner factors comprising inconsistent application of practice guidelines due to a limited database of clinical trials of effective therapies in African Americans and other underrepresented minorities; and 3) barriers in the healthcare delivery system resulting in poor access to care. The Morehouse School of Medicine Community Physicians' Network (CPN) was established to address disparities in health care by focusing on provider-specific strategies. OBJECTIVES: To: 1) use disease-specific registries to identify treatment gaps and facilitate quality improvement processes among CPN practices; 2) develop practice specific and guideline-based educational messages to promote quality care; 3) engage and train CPN-physicians for participation in approved NIH, other government, and industry-supported clinical protocols; and 4) develop a data repository of all CPN-sponsored clinical trials that include significant numbers of African Americans and other underrepresented minorities. METHODS: The disease specific outpatient registries will have the following features: 1) data structures and data elements will use standard database codes and a data dictionary; 2) HIPPA-compliant data abstraction and data transfer tool; 3) baseline chart review to establish practice patterns and provide practice specific feedback; 4) annual update of registry; 5) data registry and repository maintained on Morehouse School of Medicine's secure servers; 6) registry publications will include only aggregate data, without identification of contributing practices; 7) an electronic medical records platform will be encouraged as the ultimate data management tool for CPN practices. In addition, up to three continuing medical education (CME) programs each year will feature national speakers and promote evidence-based practice guidelines. RESULTS: Eighty five primary care and subspecialty practices are actively enrolled in CPN with a total of 385,000 annual outpatient visits. The makeup of insurance status is: HMO/PPO (45%); Medicare only (19%); Medicare HMO (11%); Medicare plus (8%); Medicaid (6%); Uninsured (11%). CONCLUSIONS: The Community Physicians' Network will address specific gaps in the health care of African-American and other minority patients by promoting quality care among its members and by facilitating participation in approved clinical trial protocols. The unique academic community partnership is consistent with the NIH roadmap goal of eliminating healthcare disparities. PMID- 16315392 TI - An empowerment intervention for women living with HIV and its adaptation for women with a diagnosis of breast cancer. AB - Because of improved therapeutic strategies, both human immunodeficiency virus (HIV) infection and breast cancer can be considered life-threatening illnesses that eventually become a chronic condition. Both diagnoses carry a psychological impact, stigma, body alterations, intense medical evaluations, and therapies. Both conditions are prevalent among minorities, especially women of Hispanic heritage. This article describes an empowerment intervention originally designed for women living with HIV and adapted for women with a diagnosis of breast cancer. We will describe the conceptual framework for the intervention and the rationale for the chosen groups. The Women's Empowerment Intervention Model consists of a series of six full-day workshops in which multiple biopsychosocial dimensions are explored within the group, and diverse experiential activities are carried out related to the day's topics. The workshops were modified to deal with the specific issues that women confront when diagnosed with breast cancer. This model was chosen precisely because it deals with specific aspects of healing and living with a chronic illness. We propose that HIV should be viewed as a chronic condition and compared to other conditions that affect women's lives in similar ways. PMID- 16315393 TI - Mechanism, measurement, and prevention of oxidative stress in male reproductive physiology. AB - Numerous factors influence male fertility. Among these factors is oxidative stress (OS), which has elicited an enormous interest in researchers in recent period. Reactive oxygen species (ROS) are continuously produced by various metabolic and physiologic processes. OS occurs when the delicate balance between the production of ROS and the inherent antioxidant capacity of the organism is distorted. Spermatozoa are particularly sensitive to ROS as their plasma membrane contains polyunsaturated fatty acids (PUFA), which oxidizes easily. They also lack cytoplasm to generate a robust preventive and repair mechanism against ROS. The transition metal ions that are found in the body have a catalytic effect in the generation of ROS. Lifestyle behaviours such as smoking and alcohol use and environmental pollution further enhance the generation of ROS and thus, cause destructive effects on various cellular organelles like mitochondria, sperm DNA etc. This article analyzes the detrimental effects of OS on male fertility, measurement of OS and effective ways to decrease or eliminate them completely. We have also provided information on oxidative stress in other systems of the body, which may be applied to future research in the field of reproductive biology. PMID- 16315394 TI - Mouse model of male germ cell apoptosis in response to a lack of hormonal stimulation. AB - As a prerequisite for studies using mutant mice, we established a mouse model for induction of male germ cell apoptosis after deprivation of gonadotropins and intratesticular testosterone (T). We employed a potent long acting gonadotropin releasing hormone antagonist (GnRH-A), acyline, alone or in combination with an antiandrogen, flutamide for effective induction of germ cell apoptosis in mice. Combined treatment with continuous release of acyline (3 mg/kg BW/day) with flutamide (in the form of sc pellets of 25 mg) resulted in almost the same level of suppression of spermatogenesis, as judged by testis weight and by germ cell apoptotic index, in 2 weeks as that reported for rats after treatment with 1.25 mg/kg BW Nal-Glu GnRH-A for the same time period. Within the study paradigm, the maximum suppression of spermatogenesis occurred after a single sc injection of high (20 mg/kg BW) dose of acyline with flutamide. The combined treatment resulted in complete absence of elongated spermatids. Germ cell counts at stages VII-VIII showed a significant (P < 0.05) reduction in the number of preleptotene (27.1%) and pachytene spermatocytes (81.9%), and round spermatids (96.6%) in acyline + flutamide group in comparison with controls. In fact, treatment with a single high (20 mg/kg BW) dose of acyline combined with flutamide in mice achieved same or greater level of suppression, measured by germ cell counts at stages VII-VIII, in two weeks when compared with those reported after daily treatment with Nal-Glu GnRH-A for 4 weeks in rats. Both plasma and testicular T levels were markedly suppressed after administration of acyline alone either by miniosmotic pump or by a single sc injection. Addition of flutamide to acyline had no discernible effect on plasma or intratesticular T levels when compared with acyline alone. These results demonstrate that optimum suppression of spermatogenesis through increased germ cell death is only possible in mice if total abolition of androgen action is achieved and further emphasize the usefulness of acyline + flutamide treated mice as a suitable model system to study hormonal regulation of testicular germ cell apoptosis. PMID- 16315395 TI - Ovariectomy, hindlimb unweighting, and recovery effects on skeletal muscle in adult rats. AB - INTRODUCTION: The effect of female sex hormones on skeletal muscle is unclear. Reports suggest that female sex hormones have an anabolic effect, have no effect, or are detrimental to skeletal muscle. METHODS: To amplify ovarian hormones' potential effects on skeletal muscle, 6-mo-old virgin intact (sham), ovariectomized (OVX), and ovariectomized plus estrogen (OVX + E2) rats were hindlimb unweighted (HLU) for 4 wk. To investigate estrogen's influence on the recovery process, OVX-HLU, sham-HLU and OVX + E2 HLU rats were cage-recovered for 2 wk following 4 wk of HLU. Changes in muscle mass were determined for the soleus (SOL), extensor digitorum longus (EDL), quadriceps (QUAD), gastrocnemius (GAST), tibialis anterior (TA), plantaris (PLA), and flexor carpi radialis (FCR). Dry weights were determined for GAST and TA. Peak tetanic tension was ascertained in the postural SOL and locomotor PLA. RESULTS: HLU resulted in a significant (p < 0.05) decline in mass of all muscles studied except EDL and FCR. The magnitude of atrophy resulting from suspension was similar between groups including wet and dry muscle mass normalized to bodyweight. Following HLU there were no differences between intact and OVX rats for contractile characteristics. Reloading for 2 wk resulted in recovery of wet muscle mass in intact and OVX + E2 rats for SOL, PLA, GAST, QUAD, and TA and in dry mass for GAST and TA. Reloaded OVX-HLU rats failed to recover wet muscle mass in all but the SOL and failed to recover GAST and TA dry weight. For example, PLA mass values for intact (sham) rats were 350 +/- 33, 284 +/- 41, and 346 +/- 44 mg for the control, HLU, and recovery groups, respectively. PLA values for OVX rats were 378 +/- 29, 316 +/- 30, and 325 +/- 23 mg for the control, HLU, and recovery groups. DISCUSSION: Results suggest that ovarian function influences the recovery of skeletal muscle mass following a period of reduced physical activity. PMID- 16315396 TI - Bed rest effects on human calf hemodynamics and orthostatic intolerance: a model based analysis. AB - INTRODUCTION: Microgravity-induced orthostatic intolerance continues to be a primary problem after space missions. Its etiology remains uncertain despite significant research efforts over the past years. We hypothesized that calf hemodynamic parameters (compliance and resistance) are significantly affected by 14 to 16-d head-down bed rest (simulated microgravity), and their alterations play a role in the pathogenesis of orthostatic intolerance (OI) following bed rest. METHODS: To estimate these parameters, we developed a model-based approach to quantitatively simulate calf vascular response to venous occlusion, which only necessitates measurement of plethysmography data. In this study, plethysmography data were obtained from 29 subjects before and after 14-16 d of head-down bed rest. The subjects also underwent a tilt/stand test before and after bed rest. RESULTS: Statistical analyses demonstrated an increase in calf compliance (1.87 +/- 0.08, mean +/- SE, pre-bed rest; 2.16 +/- 0.10, end-bed rest) but no significant change in vascular resistance following bed rest. Compared with the tilt-intolerant subjects, those who were tilt-tolerant before bed rest had significantly higher calf compliance [2.00 +/- 0.09 (tolerant); 1.58 +/- 0.09 (intolerant)] and higher vascular resistance [7.79 +/- 0.18 (tolerant); 6.91 +/- 0.40 (intolerant)]. After bed rest, no such difference was detected. DISCUSSION: Based on these results, we validated the hypothesis that, instead of causing orthostatic intolerance, higher calf compliance before bed rest leads to recruitment of compensatory mechanisms (validated by the enhanced vascular resistance during venous occlusion) for a better toleration of orthostatic stress. With the absence of orthostatic challenge during bed rest, the difference in calf hemodynamic parameters is attenuated between the tilt-tolerant and tilt intolerant groups. PMID- 16315397 TI - Legal implications of on-the-job fatigue: can employers limit outside employment? PMID- 16315398 TI - Research from the National Institutes of Health, National Institute of Diabetes and Digestive Kidney Disease, and Charles R. Drew University of Medicine and Science High School Student Research Program--Summer 2004. PMID- 16315399 TI - Abstracts of the 25th Annual Meeting of the Society for Clinical Trials, 23-26 May 2004, New Orleans, Louisiana, USA. PMID- 16315401 TI - Abstracts of the 26th Annual Meeting of the Society for Clinical Trials, 22-25 May 2005, Portland, Oregon, USA. PMID- 16315400 TI - [significance of PTH (1-84) assay in the management of chronic renal failure and the current status and future of the treatment of renal osteopathy]. PMID- 16315402 TI - [Abstracts of the 80th Annual Meeting of the French Society of Surgical Orthopedics and Trauma]. PMID- 16315403 TI - Abstracts of the XI International Workshop on CLL, 16-18 September 2005, New York, USA. PMID- 16315405 TI - Abstracts of the Global Congress of Minimally Invasive Gynecology, 34th Annual Meeting of the American Association of Gynecologic Laparoscopists, Chicago, Illinois, USA, November 9-12, 2005. PMID- 16315404 TI - Abstracts of the European Academy of Childhood Disability Annual Meeting, 19-22 November 2005. PMID- 16315406 TI - Abstracts of the joint meeting of the Physiological Society, Scandinavian Physiological Society, and the German Physiological Society. 15-19 March 2002, Tubingen, Germany. PMID- 16315407 TI - Abstracts of the 13th Annual Meeting of the Canadian Society for Exercise Physiology, November 9-12, 2005, Gatineau, Quebec, Canada. PMID- 16315409 TI - Society for Leukocyte Biology list of members. PMID- 16315408 TI - Abstracts of the 38th Annual Meeting of the Society for Leukocyte Biology, September 21-24, 2005, Oxford, England. PMID- 16315410 TI - ISHP York 2005. Airway Diseases: A Shared Approach to Management in conjunction with the 6th International Symposium on Human Pepsin, York, United Kingdom, April 6-8, 2005. Abstracts. PMID- 16315411 TI - Abstracts of the 99th Annual Scientific Assembly of the Southern Medical Association, November 10-13, 2005, San Antonio, Texas, USA. PMID- 16315412 TI - Abstracts of the 7th Congress of the European Association for Clinical Pharmacology and Therapeutics, June 25-29, 2005, Poznan, Poland. PMID- 16315413 TI - [Abstracts of the XV Annual Congress of the SAMIG (Argentine Society of General Internal Medicine), 28-29 October 2005, Buenos Aires, Argentina]. PMID- 16315414 TI - Pneumonia LOS. PMID- 16315416 TI - Sepsis and Multiorgan Dysfunction. Abstracts of the 2nd International Congress of the German Sepsis Society, September 7-10, 2005, Weimar, Germany. PMID- 16315415 TI - [Abstracts of the XX Annual Meeting of Applied Research in Chagas Disease and the VIII Annual Meeting of Applied Research in Leishmaniasis. Uberaba, Brazil, 22-24 October 2004]. PMID- 16315417 TI - The American Society for Histocompatibility and Immunogenetics 31st Annual Meeting, October 17-21, 2005, Washington, DC, USA. Abstracts. PMID- 16315418 TI - [Abstracts of the 22nd Annual Meeting of the Brazilian Society of Odontology Research, 3-7 September 2005, Aguas de Lindoia, SP, Brazil]. PMID- 16315419 TI - Frontiers in Cancer Prevention Research. Abstracts of the 4th AACR International Conference, October 30-November 2, 2005, Baltimore, Maryland, USA. PMID- 16315420 TI - [Abstracts of the Clinical Research Club of Quebec 47th annual meeting, Bromont, Quebec, Canada, September 29-October 1, 2005]. PMID- 16315421 TI - Abstracts of the 7th European Congress of Chemotherapy and Infection, October 19 22, 2005, Florence, Italy. PMID- 16315422 TI - Abstracts of the 1st Central European Forum for Microbiology (CEFORM) and the annual meeting of the Hungarian Society for Microbiology, October 26-28, 2005, Keszthely, Hungary. PMID- 16315423 TI - Abstracts from the 28th Congress of the Scandinavian Society of Anaesthesiology and Intensive Care Medicine, Reykjavik, Iceland. PMID- 16315424 TI - 2005 APA Silver Awards: Silver Achievement Awards. Police Organization Providing Peer Assistance (POPPA), New York City--helping police officers cope with on-the job pressure. PMID- 16315425 TI - World Health Assembly concludes: adopts key resolutions affecting global public health. PMID- 16315426 TI - Modern methods of treating hip fractures. AB - Contemporary surgical and anaesthetic methods should enable almost all hip fractures to be treated surgically. Many of the implants used for the internal fixation of hip fractures or the arthroplasties for replacement of the hip joint have been in use for over 50 years. Marked improvements in surgical technique have led to lower fracture and surgical complications. Undisplaced intracapsular fractures and displaced fractures in the younger patient are best treated by internal fixation. For the elderly with a displaced intracapsular fracture then hemiarthroplasty is generally used. There is insufficient evidence that total hip replacements or bipolar hemiarthroplasties have any advantage over the more traditional designs. For trochanteric fractures the sliding hip screw remains the implant of choice, although the newer intramedullary nails are valuable for more specific fracture types and their use will become more common. Following surgery almost all patients should be allowed to mobilize without any restrictions on weight bearing or hip movements. PMID- 16315427 TI - Coordinated multidisciplinary rehabilitation after hip fracture. AB - PURPOSE: To review the topic of coordinated multidisciplinary rehabilitation after hip fracture from a research perspective and to provide information to guide the provision of rehabilitation services for patients with hip fracture. METHODS: Literature review including searches of Medline, Embase, Cochrane Collaboration and evidence based clinical guidelines, checking of references of publications and consultation with researchers. RESULTS: The research evidence is heterogeneous and remains inclusive. Programs that assist patients with hip fracture to regain function and return home as soon as feasible are likely to be effective as they appear to increase the percentage of patients who return home and remain there after hip fracture. Rehabilitation programs that achieve this are likely to be cost effective. These programs involve health professionals from multiple disciplines (nurses, allied health professionals and medical practitioners) who work collaboratively, may operate in several settings, and routinely provide specific treatments that are supported by strong evidence of effectiveness. CONCLUSIONS: Patients with hip fracture should be offered a coordinated a multidisciplinary rehabilitation program with the specific aim of regaining sufficient function to return to their pre-fracture living arrangements. PMID- 16315428 TI - Leadership alert: methods for reaching out to administrators, part I. PMID- 16315429 TI - Mentoring: growth for both participants. PMID- 16315430 TI - Accessing the internet--tips for hospital librarians. Researching herbs and nutritional supplements on the Internet. PMID- 16315431 TI - Report on a benchmarking project. PMID- 16315432 TI - Tips on preparing signs and posters. PMID- 16315433 TI - Why write? PMID- 16315434 TI - Project Bookmobile takes library to nurses. PMID- 16315435 TI - Librarian recalls early days of medical librarianship. PMID- 16315436 TI - A syringe in the hand is worth two birds in the bush. PMID- 16315437 TI - DHEA supplementation: the claims in perspective. AB - Deficiency of dehydroepiandrosterone (DHEA) is associated with lupus erythematosus, diabetes mellitus, Alzheimer disease, and some cancers, but we are not yet ready to conclude that prescribing supplemental DHEA is helpful in these or any other conditions. DHEA shows some promise in observational clinical studies and laboratory experiments, but we still need large-scale human studies to answer key questions. For now, we do not have enough evidence to recommend routine treatment with DHEA. As with other supplements, quality control is always a concern, and different brands may contain different amounts of active ingredient. PMID- 16315438 TI - Safety of statins: effects on muscle and the liver. AB - Although statin drugs can have adverse effects on muscles and the liver, these effects are uncommon. Caution is warranted in patients at higher risk, ie, those who are elderly, frail, or small; have multisystem disease; are receiving immunosuppressive drugs or other medications that interact with statins; or are receiving higher doses of a statin. PMID- 16315439 TI - Hepatitis C infection: a systemic disease with extrahepatic manifestations. AB - Mixed cryoglobulinemia, renal syndromes, lymphoproliferative disorders, Sjogren syndrome, porphyria cutanea tarda, and neuropathies are all strongly associated with hepatitis C virus (HCV) infection. Diabetes, thyroid disease, and the presence of autoantibodies in the serum are also linked to HCV, but less strongly. The pathophysiologic basis for most of these syndromes seems immunologic. Cirrhosis and chronic HCV infection seem to be risk factors. PMID- 16315440 TI - A 73-year-old woman with chronic nonhealing cellulitis. PMID- 16315441 TI - Anemia in chronic heart failure: can EPO reduce deaths? AB - Many patients with chronic heart failure also have anemia, an association that has been increasingly recognized in recent years. Whether treating anemia will improve outcomes in patients with heart failure has yet to be determined, however. The decision to use an agent to treat anemia in heart failure should be made on a case-by-case basis. PMID- 16315442 TI - What is a 'failure' of bisphosphonate therapy for osteoporosis? AB - Assessing the effectiveness of bisphosphonate therapy is problematic. Bone mineral density and markers of bone turnover are often used, but the true measure is prevention of new fractures. PMID- 16315443 TI - Influenza 2005-2006: vaccine supplies adequate, but bird flu looms. AB - Influenza vaccine supplies appear to be adequate for the 2005-2006 season, though delivery has been somewhat delayed. However, in the event of a pandemic of avian flu-considered inevitable by most experts, although no one knows when it will happen-the United States would be woefully unprepared. PMID- 16315444 TI - A 39-year-old man with sudden onset of chest pain. PMID- 16315445 TI - Pulmonary embolism imaging. PMID- 16315446 TI - A psychophysical forward masking comparison of longitudinal spread of neural excitation in the Contour and straight Nucleus electrode arrays. AB - The objective of the study was to compare the widths of forward masking profiles in subjects implanted with the Nucleus 24 Contour or straight electrode array. The Contour array is typically positioned closer to the modiolus than the straight array. Subjects were fourteen postlingually hearing-impaired adults with severe-profound hearing loss, seven used the Contour array and seven used the straight array. Forward masking profiles were measured at three positions along the array (apical, mid, and basal) using maskers at the 15% loudness level. It was hypothesized that masking profile widths would be more sensitive to differences in distance from the neural structures using low-level maskers. Masking width was calculated at the 50% point of the masking peak amplitude. There were no significant differences in masking widths between Contour and straight array subject groups. Current levels for hearing thresholds and maximum comfortable listening levels were significantly lower for the Contour array subjects. PMID- 16315447 TI - Auditory steady-state responses (ASSR): effects of modulation and carrier frequencies. AB - Presented are results relevant to extending the utility of the auditory steady state response (ASSR) in threshold estimation at high-frequency carriers and to the accuracy of thresholds estimated using modulation frequencies near 40 versus 80 Hz. Initially, efforts were directed at confirming various findings reported in the literature apropos effects of several basic ASSR parameters. Results supplement others' observations suggesting that ASSR detection limits overestimate behavioral thresholds for conventional audiometric (carrier) frequencies from 500 to 4000 Hz. Further investigation revealed even greater errors of threshold estimates for 8000 and 12000 Hz, by about 14 and 22 dB on average, respectively. Although suggesting high-frequency ASSR testing to be efficacious, technical advances and additional work is needed to establish clinical utility. Comparison of effects of modulation frequency suggested ASSR thresholds with 40 Hz modulation to fall closer to behavioral threshold than those estimated at 80 Hz. Consequently, when circumstances permit, 40 Hz ASSR testing may be the method of choice (e.g. in the assessment of malingers, who may be tested awake/alert). PMID- 16315448 TI - Speech understanding in quiet and noise, with and without hearing aids. AB - Speech recognition and cognitive functions important for speech understanding were evaluated by objective measures and by scores of perceived effort, with and without hearing aids. The tests were performed in silence, and with background conditions of speech spectrum random noise and ordinary speech. One young and one elderly group of twelve hearing-impaired subjects each participated. Hearing aid use improved speech recognition in silence (7 dB) and in the condition with speech as background (2.5 dB S/N), but did not change the perceived effort scores. In the cognitive tests no hearing aid benefit was seen in objective measures, while there was an effect of hearing aid use in scores of perceived effort, subjects reported less effort. There were no age effects on hearing aid benefit. In conclusion, hearing aid use may result in reduced effort in listening tasks that is not associated with improvement in objective scores. PMID- 16315449 TI - Predictive factors for development of severe tinnitus suffering-further characterisation. AB - The purpose of this study was to identify risk factors for Severe Tinnitus Suffering (STS) and to describe the results from psychiatric diagnostic interviews of STS positive and STS negative patients. Consecutive tinnitus patients (n = 127) completed the Nottingham Health Profile (NHP) and the Tinnitus Severity Questionnaire (TSQ) and answered general health questions at the first visit to the clinic and at eighteen months follow-up. Twenty-four months after the first tinnitus consultation, standardized diagnostic interviews (SCID-P), according to DSM-IIIR, were performed by an experienced psychiatrist. The calculated probability for STS was 93%, if three items from the NHP (18,32,33) were positive. STS positive patients had more anxiety disorders with more psychosocial stress and lower global assessment of function scores. Results suggest that the NHP may be a useful tool in the clinical management of tinnitus patients and emphasises the importance of identifying depressive and anxiety disorders in this population. PMID- 16315450 TI - Predisposing factors for inner ear hearing loss association with chronic otitis media. AB - The aim of the present study was to investigate the consequences of chronic otitis media on inner ear function. Retrospective analysis of conventional pure tone audiometry tests was carried out on 344 patients who were scheduled for surgical treatment of unilateral chronic otitis media without other risk factors for sensorineural hearing loss. Bone conduction thresholds of diseased ears were compared with those of contralateral, non-diseased ears. Selected clinical features were assessed among diseased ears to examine possible influences on inner ear function. Mean bone conduction threshold differences varied from 0.6 dB at 0.5 kHz to 3.7 dB at 4 kHz. These differences augmented with increasing duration of middle ear disease. Impaired hearing by bone conduction thresholds of diseased ears correlated with increased age at every frequency and with an interruption of the ossicular chain only at higher frequencies. The severity of sensorineural hearing loss correlated with longer duration of middle ear disease. Thus, surgical treatment of dry and apparently stable tympanic membrane perforation is warranted. PMID- 16315452 TI - The road not taken. PMID- 16315451 TI - Dead regions and noisiness of pure tones. AB - Some hearing-impaired subjects report pure tones as sounding highly distorted and noise-like. We assessed whether such reports indicate that the tone frequency falls inside a dead region (DR). Nine hearing-impaired and four normally hearing subjects rated pure tones on a scale from 1 to 7, where 1 indicates clear tone and 7 indicates noise. A white noise was presented as a reference for a sound that should be rated as 7. Stimuli covered the whole audible range of frequencies and levels. The noisiness ratings were, on average, higher for hearing-impaired subjects than for normally hearing subjects. For the former, the ratings were not markedly different for tones with frequencies just outside or inside a DR. However, ratings always exceeded 3 for tones falling more than 1.5 octaves inside a DR. The results indicate that judgement of a tone as sounding noise-like does not reliably indicate that the tone frequency falls in a DR. Both normally hearing and hearing-impaired subjects rated 0.125 kHz and 12 kHz tones as somewhat noise-like, independently of the existence of a DR. PMID- 16315453 TI - Risk and ontology. PMID- 16315454 TI - TennCare crisis an open door to comprehensive care. PMID- 16315455 TI - Volunteer physician liability protection? PMID- 16315456 TI - Important TennCare Changes. PMID- 16315457 TI - Meth in Tennessee: fighting the "devil's drug". PMID- 16315458 TI - Small town practice: caring for "Family". PMID- 16315460 TI - The greatest job. PMID- 16315459 TI - On being a doctor...where I'm from. PMID- 16315461 TI - Maintaining the idea. PMID- 16315462 TI - Technique in question. PMID- 16315463 TI - Playing "Russian roulette" with inhalants, pt. 2. PMID- 16315464 TI - Needs-based rural medical education training. PMID- 16315465 TI - The goose that laid the golden capsule, part III. PMID- 16315466 TI - The renal patient with cardiovascular disease--no longer a simple plumbing problem. AB - The clinical syndrome of chronic kidney disease (CKD) with coronary artery disease (CAD) is a clinical challenge. The risk of cardiovascular disease in patients with renal impairment appears to be far greater than in the general population. Despite the high prevalence of CKD related to cardiovascular disease (CVD), it remains understudied. Most of the current research comes from small community-based studies and retrospective reviews, assuming that patients with CKD will similarly benefit from treatments as patients with normal renal function. Most of the current clinical trials have excluded patients with CKD even though they represent a group of people at high risk for cardiovascular (CV) complications. CKD is associated with adverse CV outcomes and higher mortality even after adjustment for conventional risk factors. Declining renal function portends increasing CV risk and may be explained by several other factors that are being investigated, including inflammation, decreased vascular compliance, homocysteine, albuminuria and dyslipidemia. In addition, there is a lack of appropriate intervention in patients with CKD, despite established awareness of their high cardiovascular risk. PMID- 16315467 TI - What will you say about HPV? PMID- 16315468 TI - Suicide and the military. PMID- 16315469 TI - Into the abstract. PMID- 16315470 TI - Evaluating human factors in augmented reality systems. PMID- 16315471 TI - Designing a visualization framework for multidimensional data. PMID- 16315472 TI - Visualization of fluid mixing in microchannels. PMID- 16315473 TI - Moving mixed reality into the real world. PMID- 16315474 TI - Mixed reality in education, entertainment, and training. PMID- 16315475 TI - Using augmented reality to treat phobias. PMID- 16315476 TI - Pedestrian Tracking with shoe-mounted inertial sensors. PMID- 16315477 TI - Augmented reality projects in the automotive and aerospace industries. PMID- 16315478 TI - High-dynamic-range still-image encoding in JPEG 2000. PMID- 16315479 TI - Designing graphics programming interfaces for mobile devices. PMID- 16315480 TI - How to solve a quadratic equation. PMID- 16315481 TI - Dance movement: a focus on the technology. PMID- 16315482 TI - Preparation of proteins from different organs of Japanese morning glory by an in vivo electro-elution procedure. AB - An electro-elution procedure has been employed efficiently to collect proteins from stem segments, young leaves and roots of the Japanese morning glory. Electrophoretic conditions were optimised, including the size of segments (10-30 mm), the strength of the current for electro-elution (2.5-10 mA), and the exposure time of electro-elution (2-12 h). From the same quantity of organs, the in vivo electro-elution procedure permitted the collection of an amount of protein up to six times greater than that obtained with an earlier-reported centrifugation procedure. Both preparations were analysed by SDS-PAGE and showed similar protein profiles. This new technique provided an interesting insight into the large differences in both the quality and quantity of proteins between different organs of the plants. The average amount of protein collected from organs was 0.1 mg/g of tissue fresh weight. It is expected that this procedure may facilitate the discovery of new proteins with unique functions in extracellular matrices involved in the response of plants to various external stimuli. PMID- 16315483 TI - Speciation analysis and the assessment of bioavailability of manganese in phytomedicines by extraction with octanol and determination by flame atomic absorption spectrometry. AB - Trace elements in phytomedicines are present in the form of metallic complexes. Since n-octanol, a long-chain alkanol, presents a configuration similar to that of carbohydrates and lipids, the lipophilicity and absorptivity of organic medicines may be assessed from their distribution coefficients between octanol and water. This strategy has been used in order to define the species of manganese in a number of phytomedicines and to study the distribution of manganese in decoctions of phytomedicines in the stomach and the intestine. The concentrations of manganese in the original herbal materials and in octanol- and water-soluble fractions were determined by flame atomic absorption spectrometry following mixed acid digestion. The acidities of gastric and intestinal juices, the phytomedical composition and the compatibility of phytomedicines, i.e. the combination of single phytomedicines, greatly affected the manganese complexing ligands and determined the species and bioavailability of manganese. It is concluded that a knowledge of the level of octanol-soluble manganese in a phytomedicine could form the basis of dosage design in order to avoid manganese overload. PMID- 16315484 TI - Purification of betulinic acid from Eugenia florida (Myrtaceae) by high-speed counter-current chromatography. AB - A high yield of betulinic acid (up to 17% from the ethanolic extract) was found in the leaves of Eugenia florida collected in south-eastern Brazil, making this species a potential commercial source of the title compound. Extracts of E. florida were subjected to solvent partition, and rapid high-speed counter-current chromatography (HSCCC) was applied to the semi-crude extracts to afford betulinic acid in high purity. The mobile and stationary phases were derived from the two phase solvent system composed of n-hexane-ethyl acetate-methanol-water (10:5:2.5:1). The developing solvent system (stationary and mobile phases) for optimum HSCCC separation was chosen by dissolving the fraction to be chromatographed in the proposed solvent mixture and determining the amount of betulinic acid in each phase by densitometric TLC. Purified betulinic acid was characterized by 13C-NMR, GC-MS and co-injection of its methyl ester with standards in GC-FID. The HSCCC technique is commonly employed to isolate triterpene glycosides, but is applied in this study to an aglycone. PMID- 16315485 TI - Quantification of Aconitum alkaloids in aconite roots by a modified RP-HPLC method. AB - The three Aconitum alkaloids, aconitine (1), mesaconitine (2) and hypaconitine (3), are pharmacologically active but also highly toxic. A standardised method is needed for assessing the levels of these alkaloids in aconite roots in order to ensure the safe use of these plant materials as medicinal herbs. By optimising extraction, separation and measurement conditions, a reliable, reproducible and accurate method for the quantitative determination of all three Aconitum alkaloids in unprocessed and processed aconite roots has been developed. This method should be appropriate for use in the quality control of Aconitum products. The three Aconitum alkaloids were separated by a modified HPLC method employing a C18 column gradient eluted with acetonitrile and ammonium bicarbonate buffer. Quantification of Aconitum alkaloids, detected at 240 nm, in different batches of samples showed that the content of 1, 2 and 3 varied significantly. In general, the alkaloid content of unprocessed roots was higher than that of processed roots. These variations were considered to be the result of differences in species, processing methods and places of origin of the samples. PMID- 16315486 TI - HPLC coupled on-line to ESI-MS and a DPPH-based assay for the rapid identification of anti-oxidants in Butea superba. AB - A reversed-phase HPLC coupled on-line to a radical scavenging detection system and MS/MS was developed in order to combine separation, activity determination and structural identification of anti-oxidants in complex mixtures in one run. The sample was separated by HPLC and the eluate split into two flows. The major portion was fed into an electrospray ionisation MS/MS system, while the minor part was mixed with a free radical, 2,2'-diphenyl-1-picrylhydrazyl (DPPH), and the reaction determined spectrophotometrically. The negative peaks, which indicated the presence of anti-oxidant activity, were monitored by measuring the decrease in absorbance at 517 nm. The developed method was successfully applied to the identification of anti-oxidant compounds in a fraction, obtained by solid phase extraction, of an extract of a Thai medicinal plant, Butea superba Roxb. The anti-oxidant compounds were separated and identified as procyanidin B2, (-) epicatechin and procyanidin B5. PMID- 16315487 TI - On-line identification of unstable iridoids from Jamesbrittenia fodina by HPLC-MS and HPLC-NMR. AB - HPLC-UV-MS analysis of the methanol extract of Jamesbrittenia fodina (Wild) O. M. Hilliard (Scrophulariaceae) revealed the presence of different iridoid cinnamic esters; however, isolation of these constituents was prevented by instability problems. HPLC-UV-MS and HPLC-NMR analysis of the mixtures obtained after a tentative isolation indicated that, in the first instance, instability was due to a light-induced cis/trans isomerisation of the cinnamic moieties. Further investigation of related compounds showed an additional instability problem linked to other chemical transformations. A detailed HPLC-NMR-MS study of these fractions demonstrated that the modifications occurred on the rhamnose moiety of these iridoids. It could be concluded that the second type of instability was attributable to transesterification of the cinnamic moiety on the rhamnose unit. The recording of stop-flow HPLC-NMR spectra for specific HPLC peaks permitted the direct monitoring of these transformations. Based on these on-line data, six new unstable aucubin derivatives were efficiently characterised. PMID- 16315488 TI - Determination of cardamonin using a chemiluminescent flow-injection method. AB - A sensitive and selective flow injection chemiluminescence method for the determination of cardamonin over the range 1.0 x 10(-8) to 8.0 x 10(-6) g/mL is described. The method is based on the enhancement by cardamonin of the chemiluminescence of the reaction between cerium (IV) and rhodamine 6G in sulphuric acid medium. The optimised flow injection procedure yielded a detection limit for cardamonin of 8.8 x 10(-9) g/mL, whilst the relative standard deviations of intraday and inter-day precision were below 2.5%. The method has the advantages of high sensitivity and a wide linear range. It was successfully applied to the determination of cardamonin in Alpinia katsumadai Hayata. The mechanism of the chemiluminescence reaction is proposed. PMID- 16315489 TI - Fingerprinting of Cretaceous higher plant resins by infrared spectroscopy and gas chromatography coupled to mass spectrometry. AB - Ambers from Penacerrada (Basque Country, North Spain) have been analysed by IR spectroscopy and GC-MS in order to study the polymeric and solvent-soluble GC amenable fractions, respectively. All samples showed branched monoalkybenzenes, bicyclic sesquiterpenoids and tricyclic diterpenoids related to pimaric acids precursors, suggesting a paleobotanic origin from araucarian species. Molecules containing oxygenated polar functionalities were not found, which is in agreement with the Cretaceous origin of the samples and in good correspondence with the IR spectra and with the low oxygen content estimated from elemental composition analysis. In addition, no IR bands for exocyclic double bonds and very few GC amenable monounsaturated molecules were found, which provides further evidence of the loss of olefinic groups. These features show that the age-dependent loss of oxygenated and olefinic functions is a general process affecting the whole amber structure, not only the solvent soluble fraction. PMID- 16315490 TI - Characterisation of the phenolic profile of Boerhaavia diffusa L. by HPLC-PAD MS/MS as a tool for quality control. AB - Phenolic acids and flavonols of nine leaf and three root samples of Boerhaavia diffusa L., collected at different locations and subjected to several drying procedures, were characterised by reversed-phase HPLC-PAD-ESI/MS for the first time. Ten phenolic compounds were identified: 3,4-dihydroxy-5-methoxycinnamoyl rhamnoside, quercetin 3-O-rhamnosyl(1-->6)galactoside (quercetin 3-O robinobioside), quercetin 3-O-(2"-rhamnosyl)-robinobioside, kaempferol 3-O-(2" rhamnosyl)-robinobioside, 3,5,4'-trihydroxy-6,7-dimethoxyflavone 3-O-galactosyl(1 ->2)glucoside [eupalitin 3-O-galactosyl(1-->2)glucoside], caffeoyltartaric acid, kaempferol 3-O-robinobioside, eupalitin 3-O-galactoside, quercetin and kaempferol. Quantification was achieved by HPLC-PAD and two phenolic patterns were found for the leaves, in which quercetin 3-O-robinobioside or quercetin 3-O (2"-rhamnosyl)-robinobioside was the major compound. Caffeoyltartaric acid was only present in the root material where it represented the main phenolic constituent. The results obtained demonstrated that the geographical origin (particularly the nature of the soil), but not the drying process, influences the phenolic composition. PMID- 16315491 TI - HPLC comparison of supercritical fluid extraction and solvent extraction of coumarins from the peel of Citrus maxima fruit. AB - The efficiency of carbon dioxide supercritical fluid extraction (SFE) of the biologically active compounds imperatorin, meranzin and meranzin hydrate from the fruit peel of Citrus maxima Merr. has been compared with that of solvent extraction with acetone. Under the best SFE conditions tested for the three coumarins, which involved extraction at 50 degrees C and 27.6 MPa, the extractive efficiencies were 84, 76 and 18% for imperatorin, meranzin and meranzin hydrate, respectively. The presence of modifiers significantly affected the extraction efficiency: the highest extraction efficiency of the three coumarins was obtained with ethanol as modifier. PMID- 16315492 TI - Analysis of macamides in samples of Maca (Lepidium meyenii) by HPLC-UV-MS/MS. AB - The macamides are a distinct class of secondary metabolites that have so far been found only in Lepidium meyenii Walp. (Maca). Using HPLC-UV-MS/MS, the main macamides have been identified as n-benzylhexadecanamide, n-benzyl-(9Z) octadecenamide, n-benzyl-(9Z, 12Z)-octadecadienamide, n-benzyl-(9Z, 12Z, 15Z) octadecatrienamide and n-benzyloctadecanamide. The identities of n-benzyl-(9Z) octadecenamide and n-benzyl-(9Z, 12Z)-octadecadienamide were confirmed by comparison of chromatographic and spectral properties with synthetic analogues. Total macamides have been quantified by HPLC-UV in plant material from different vendors using n-benzylhexadecanamide as an external standard. The amount of macamides in the dried plant material ranged from 0.0016 to 0.0123%. PMID- 16315493 TI - HPLC-MS/MS analysis of willow bark extracts contained in pharmaceutical preparations. AB - Preparations containing willow bark extract are popular herbal remedies, but they are mostly standardised with respect to only one compound (usually salicin). RP HPLC using a C18-column eluted with water:methanol:tetrahydrofuran and coupled to electrospray triple-quadrupole MS and MS/MS was used for the characterisation of dried extracts of Salix spp. and for the identification of their constituents. Comparison with reference substances led to the identification of 13 compounds (saligenin, salicylic acid, salicin, isosalicin, picein, salidroside, triandrin, salicoylsalicin, salicortin, isosalipurposide, salipurposide, naringenin-7-O glucoside and tremulacin). Two pharmaceutical preparations containing willow bark extract, used in clinical trials and labelled Salix daphnoides and S. purpurea x daphnoides extracts, were compared using the described method and exhibited several clear differences, the most prominent of which was the possible presence of picein in the former preparation. The described method may be utilised for the characterisation of herbal medicines in order to ensure comparability of medication in further clinical trials. PMID- 16315494 TI - Phytochemical profiling of New and Old World Hypericum (St. John's Wort) species. AB - Botanical extracts of Hypericum perforatum L. (common St. John's Wort) are used in the USA and in Europe as a treatment for mild to moderate depression, although controversy surrounds the identity of the active constituent(s). RP-HPLC with photodiode array detection was used to separate and quantify nine compounds of pharmacological interest in extracts from 74 taxa of Hypericum native to the Old and New World. Chemical profiles of these constituents may be used to distinguish extracts of H. perforatum from those of other species of Hypericum, and to indicate species that may be of interest for further phytochemical investigation. PMID- 16315495 TI - Current awareness in phytochemical analysis. PMID- 16315496 TI - Providing a forum for PA authors practicing in surgery. PMID- 16315497 TI - A comprehensive approach primary headaches. PMID- 16315498 TI - Evaluation and management of peripheral edema. PMID- 16315499 TI - Endoscopic vessel harvesting in coronary artery bypass graft surgery. PMID- 16315500 TI - Advances in treating early breast cancer. PMID- 16315501 TI - Multiple "warts" on the lower legs of a 52-year-old woman. PMID- 16315502 TI - Case of the month. Diagnostic callenges from your case files. Bilateral central serous chorioretinopathy (CSCR). PMID- 16315504 TI - Addressing design for maintainability. PMID- 16315503 TI - Contemporary treatment options for overactive bladder. PMID- 16315505 TI - Insight into process steam issues. PMID- 16315506 TI - Long-term planning can unlock benefits. AB - Is short-term thinking still preventing UK hospitals from procuring surgical facilities that could offer greater flexibility and improved patient outcomes over time? Modular theatre supplier Maquet thinks so. PMID- 16315507 TI - Tuning in to the way ahead. AB - The airwaves bring Nicholas Marshall thoughts on the direction that design of healthcare premises should be taking. PMID- 16315508 TI - Sensor accuracy is clinically essential. PMID- 16315509 TI - Multi-benefit course launched. AB - To help reduce the large number of accidents and MRSA infections that arise in the NHS annually, the Institution of Occupational Safety and Health (IOSH) has launched a training course for all healthcare staff. Health Estate Journal reports. PMID- 16315510 TI - Anti-ligature design reduces patient risk. AB - Advice for facilities managers considering the removal of ligature points to reduce the risk of patient suicide and self-harm is provided by Laidlaw Solutions. PMID- 16315511 TI - Spreading the word on infection control. AB - Jason Rayfield reports from the Seventh Annual Infection Control Nurses Association Conference, where education, surveillance and the threat posed by emerging bacterial strains were high on the discussion agenda. PMID- 16315512 TI - Full-service community schools: a strategy--not a program. AB - The concept that drives the emerging full-service community school movement is this: Schools cannot address all the problems and needs of disadvantaged children, youth, and families. Community schools are operated jointly by school systems and community agencies, are open extended hours, and may provide the site for after-school programs, primary-care health services, mental health counseling, parent education and involvement, and community development. No two community schools are alike. They grow out of a planning process that involves all stakeholders, school personnel, community-based organizations, city and county government, parents, and students. The Quitman Street Community School in Newark, New Jersey, exemplifies this approach. PMID- 16315513 TI - The Children's Aid Society community schools: a full-service partnership model. AB - In 1989, the Children's Aid Society (CAS) created an unprecedented partnership with the New York City Board of Education by developing a comprehensive response to the pressing needs of children and families in the northern Manhattan neighborhood of Washington Heights. After three years of careful planning, CAS and the New York City public schools opened the first community school at Intermediate School 218, offering a full array of supports, services, and learning opportunities. Adding, on average, one partnership school per year and remaining very flexible in adapting its model to the individual needs of each community, CAS now has thirteen community schools around New York City. The model's flexibility is seen also in the success of its national and international adaptation-an intentional part of CAS's work. PMID- 16315514 TI - The New York City Beacons: rebuilding communities of support in urban neighborhoods. AB - Established in 1991 in New York City and now operating in at least seven other cities, Beacons are designed to rebuild communities of support for children and youth in urban neighborhoods. The Beacon framework is based on research findings and practitioner experience indicating that programs taking a youth development approach are more effective than those focused on "fixing" specific youth problems. Successful Beacon programs provide positive ways to meet young people's need for safety, a sense of belonging, and mastery; they also provide opportunities for decision making and contributing to others. There are currently eighty Beacons in New York City, serving about 140,000 youth and adults annually. Beacons have been replicated in several parts of the country, including Denver, Minneapolis, Oakland, Palm Beach County, Philadelphia, San Francisco, and Savannah. The Youth Development Institute of the Fund for the City of New York provides technical assistance and training to Beacons in New York City and in all seven replication sites. PMID- 16315515 TI - University-assisted community school program of West Philadelphia: democratic partnerships that make a difference. AB - The university-assisted community school model is showing results for children and youth in West Philadelphia. The University of Pennsylvania's (Penn's) Center for Community Partnerships has coordinated universitywide efforts, in partnership with the community, in order to create and develop community school programs. The Sayre program aims to become a university-assisted community school, with a comprehensive community problem-solving curriculum and communitywide program that is fully integrated across both the Sayre curriculum and the curriculum of a number of Penn's schools. The Penn-Sayre project demonstrates that higher education can be a permanent anchor for revitalizing schools and communities if the vast resources it possesses, particularly its faculty, students, and staff, are brought to bear in a coordinated fashion. PMID- 16315516 TI - Full-service community schools: cause and outcome of public engagement. AB - Public Education Network (PEN)-a national organization of local education funds (LEFs)-along with individuals working to improve public schools and build citizen support for quality public education, embarked on an initiative to address the academic, enrichment, and social support needs of young people to ensure their success in and out of school. PEN's Schools and Community Initiative called for the active participation of broad constituencies-policymakers, stakeholders, and the public-at-large-to create and implement their common vision for full-service community schools. By engaging broad constituencies across communities, LEFs have been able to build strong relationships between and among community institutions under a common vision. PMID- 16315517 TI - Anchors of the community: community schools in Chicago. AB - In partnership with Chicago's public and private sectors, Chicago Public Schools (CPS) has successfully implemented a citywide education reform effort, designed to transform Chicago's neighborhood schools into vibrant centers of the community. Mayor Richard M. Daley and Arne Duncan, CEO of CPS, launched the Community Schools Initiative in January 2002. What started as an idea that was developed by a local foundation has now grown into the largest-scale community school effort in the nation, with sixty-seven schools in operation and a plan to move to one hundred community schools by 2007. This initiative currently involves seventeen private funders, ten technical assistance providers, thirty-four community-based organizations that offer on-site services to children and families, and over three hundred additional community partnerships that provide one-day events such as health fairs and violence prevention workshops. PMID- 16315518 TI - The school community council: creating an environment for student success. AB - A model of community-school partnerships is developing within a school district in Evansville, Indiana. Based on a full-service community school philosophy, the model started in one elementary school in the Evansville-Vanderburgh School Corporation and has expanded into a districtwide initiative called the School Community Council. The council is made up of over seventy community organizations and social service agencies working together to establish full-service schools as places of community and to enhance youth and family development. PMID- 16315519 TI - Aligning systems to create full-service schools: the Boston experience, so far. AB - Leaders of full-service schools in Boston seek to expand the number and increase the impact of Boston's full-service schools, catalyzing a realignment of public resources and an expansion of private investment. The Full-Service Schools Roundtable, led by a dynamic staff and supported by the mayor and the superintendent of schools, is a steadily growing coalition of educators, public agencies, human service providers, and community leaders. Challenges for the Roundtable are to build the public will to invest in full-service schools; secure leadership from stakeholders; share accountability across sectors so that schools prioritize youth development and health, and service providers share responsibility for school success; and become a political force, championing the strategic realignment of public investments based on child outcomes. PMID- 16315520 TI - Schools uniting neighborhoods: the SUN initiative in Portland, Oregon. AB - The SUN Community Schools Initiative is a community-driven model that allows each school community to design the programs that fit neighborhood needs in Portland, Oregon. County and city governments, local school districts, and community agencies have jointly leveraged resources to support fifty-one community schools. The program is managed by the Multnomah County Department of School and Community Partnerships. The City Parks and Recreation Bureau oversees twelve sites staffed by city employees. Strong support across political systems aligns funding and reduces the fragmentation in existing funding patterns. PMID- 16315521 TI - California's Healthy Start: A solid platform for promoting youth development. AB - A school in Los Angeles County reports absences down by 30 percent and disciplinary actions down by 10 percent. A town near Fresno reports having 99 percent of their new kindergartners ready to start school on the first day of class because their immunization and school readiness outreach was so thorough. A school in San Diego reports youth tobacco use down from 15 percent to 3 percent, absences down by 10 percent, and detentions down by over 50 percent. Schools in Humboldt report a 30 percent improvement in math scores and a 40 percent improvement in reading scores. Young adults report that the assistance they received as teens through their school's Healthy Start program saved their lives and enabled them to be successful parents today. These are results from one of California's most successful education mandates-SB620 1991-California's Healthy Start. PMID- 16315522 TI - Building the community school movement: vision, organization, and leadership. AB - On a local level, creating and sustaining community schools requires leadership from local government, schools, businesses, and nonprofit organizations. These groups must provide the fuel and direction to move the community school strategy forward along a common vision and with strategic methods for financing. At the federal level, it must continue to build constituency for community schools if it is to succeed, although the community school movement has made great strides in recent years. There is not now a coherent federal framework to support the community school vision. The proposed Full Services Community Schools legislation would build a national constituency and legislate key principles advocated by the Coalition for Community Schools: developing districtwide community school strategies, focusing on results, and improving coordination of funding streams. PMID- 16315523 TI - Our future in red tape. PMID- 16315524 TI - Could one-to-one pay bargaining lead to higher nurses' salaries? PMID- 16315525 TI - 'It should be a given that students are respected'. AB - Nursing students deserve respect. Unfortunately, they are often excluded, resented or even bullied on placements, by qualified nurses as well as other staff. This can lead to poor self-confidence or, worse, the student leaving nursing altogether. Nursing Standards Charter for Change campaign calls for nursing students to be respected by their peers. PMID- 16315526 TI - Going the distance. AB - Over the past ten years, 4,500 nurses have embarked on distance learning courses run by the RCN Institute. In addition to increased knowledge, many have gained in self-confidence. Students have ranged in age from 21 to 72. The majority have caring commitments for children or older relatives. The institute suggests that the backing of partners is essential to success. Although the majority of students are UK-based, 11 per cent are from overseas. PMID- 16315527 TI - Single rooms and patient choice. AB - AIM: To determine whether the allocation of single rooms in a large NHS trust hospital was based on clear criteria and whether patient choice was reflected in this practice. METHOD: Structured interviews were conducted with nursing staff and patients. In addition, a survey was undertaken to quantify the level of demand. RESULTS: Nursing staff were mainly influenced by the clinical needs of patients. Even in an ideal situation nearly half the staff did not mention patient choice. Approximately half of the patients would have preferred a single room, which means that demand greatly exceeds supply. Most patients did not know why they had been allocated to a particular bed. CONCLUSION: More single rooms would allow greater choice but some shared areas should be maintained. Local guidance on allocating beds would help to manage the situation more fairly. PMID- 16315528 TI - Higher level practice in community nursing: part 1. AB - Part one of this article describes the development of the nurse practitioner role in the UK. Part two reports on a research study examining whether or not community nurse practitioners were able to achieve a 'higher level of practice' as envisaged by the United Kingdom Central Council and will be published in next week's Nursing Standard. PMID- 16315529 TI - Prone positioning in patients with acute respiratory distress syndrome. AB - Prone positioning is a technique used to treat patients with acute respiratory distress syndrome (ARDS). Nurses are ideally placed to initiate and manage prone positioning. This article examines the aetiology of ARDS and how nurses can identify this condition. Patient groups that benefit most from prone positioning are considered as well as the research evidence on the use of prone positioning in the treatment of ARDS. This article addresses nurse involvement in delivering therapy and suggests that nurses alert members of the multidisciplinary team to the possible use of this therapy. PMID- 16315530 TI - A proactive approach to asthma. AB - Asthma is a common chronic inflammatory condition the incidence of which appears to be increasing in the UK. It has a substantial impact on the lives of the people it affects. A proactive approach to asthma by healthcare professionals working with patients with this condition may substantially improve asthma management. PMID- 16315531 TI - Self-harm and suicide. PMID- 16315532 TI - First impressions count. PMID- 16315533 TI - Leading light. PMID- 16315534 TI - The mother load. PMID- 16315535 TI - Liaising with the university. PMID- 16315536 TI - Oncology nurses explore new innovations in nutritional care. PMID- 16315537 TI - Surviving through resiliency. PMID- 16315538 TI - Artificial nutrition and hydration at the end of life: whose decision is it? PMID- 16315539 TI - On the front lines. PMID- 16315540 TI - The new bankruptcy code: what it means for nephrology practices. PMID- 16315541 TI - Companies look to treat metabolic syndrome despite debate over its relevance. PMID- 16315542 TI - PKD: a progress report, new directions in research. PMID- 16315543 TI - Patient group aimed at improving quality of life. PMID- 16315544 TI - The emperor has no clothes (but has a catheter): a perspective on the state of chronic kidney disease care. AB - Dialysis providers are capable and interested in participating in the care of CKD patients. But dialysis providers do not, for the most part, become involved in the patients' care until CMS Form 2728 is completed and signed by the attending nephrologist. We believe that dialysis providers are interested in broadening their role, but only if the nephrology community believes this role is appropriate and supports such a resource collaboration. If CMS wants dialysis providers to get more involved in CKD patient management, it is extremely important that this effort not be another unfunded mandate that will result in an increase in the burden on current nursing, dietary, and social work dialysis staff providing current ESRD care. If dialysis providers are to collaborate on CKD care with the nephrology community, they must staff appropriately and there must be a definitive change in the Medicare reimbursement system to support the use of CKD guidelines that will result in a healthier and less costly patient presenting to the dialysis program at the point that they reach end-stage renal disease. PMID- 16315545 TI - The Wellpoint/Anthem approach to chronic kidney disease. PMID- 16315546 TI - Preparing for nephrology practice expansion. PMID- 16315547 TI - Wellness programming: nephrology social work expands its role in renal disease management. PMID- 16315548 TI - The West Nile Virus and the dialysis/transplant patient. PMID- 16315549 TI - Thought and discussion among patients about stopping dialysis. PMID- 16315550 TI - RRI conference offers updates on CKD, research and treatment. PMID- 16315551 TI - Contraception and sexual health, 2004/05. AB - Questions on contraceptive use and sexual health were included in the Office for National Statistics (ONS) Omnibus Survey in June, September and November 2004 and March 2005. Questions were asked of women aged 16-49 and men aged 16-69. The report based on this module contains comparisons with results from the ONS Omnibus surveys 1997 to 2004. Each year approximately 4,000 adults are interviewed in Great Britain. PMID- 16315552 TI - Leading causes of death in England and Wales--how should we group causes? AB - This article examines how best to identify the leading causes of mortality in England an Wales, by using different way of grouping causes of death, based on a list developed by the World Health Organization (WHO). Four different versions of this list are compared. The leading cause of death across all age groups depends on the ways in which common diseases and external causes are aggregated or disaggregated into groups. Areas of particular debate, examined in this article, are the grouping or splitting of accidents by mechanism and cancers by site within leading cause lists. These affect which causes appear in the top ten, and their order in different age groups. PMID- 16315553 TI - Person, place or time? The effect of individual circumstances, area and changes over time on mortality in men, 1995-2001. AB - This article uses the ONS Longitudinal Study to explore, for a cohort of adult males who were aged 26 or over in 1971, the relative influence on mortality in 1995-2001 of their place of residence and individual socioeconomic circumstances, at three censuses over a 20-year period. Factors examined in this analysis include social class, neighbourhoo deprivation (at ward level), unemployment, residence in the South East region in 1971 or 1981 housing tenure, and change in social class and housing tenure between 1971 and 1991. The variation in mortality attributable to the local authority district of residence in 1991 was also investigated. PMID- 16315554 TI - Life expectancy at birth by local authorities in England and Wales, 2002-2004. PMID- 16315555 TI - Infant and perinatal mortality by social and biological factors, 2004. PMID- 16315556 TI - Mortality statistics 2003: general. PMID- 16315557 TI - The vitamin D and sunlight controversy--we will wait and see. PMID- 16315558 TI - What is your diagnosis? Graft-versus-host disease. PMID- 16315559 TI - An umbilical polyp in an infant. AB - The persistence at birth of the omphalomesenteric (vitelline) duct may result in life-threatening consequences. Early identification of this congenital anomaly is essential for prompt surgical treatment to eliminate the risk of prolapse and herniation. A neonatal umbilical polyp may indicate the presence of an omphalomesenteric duct remnant. We describe the diagnosis and surgical treatment of an infant with an umbilical polyp. We also present an overview of the diagnosis and treatment of vitelline duct remnants and their associated anomalies. PMID- 16315560 TI - The use of topical imiquimod for the treatment of actinic keratosis: a status report. AB - Topical imiquimod 5% cream is approved for the treatment of actinic keratosis (AK), superficial basal cell carcinoma, and external genital warts. The drug's mechanism of action is via stimulation of innate and acquired immune responses, which ultimately leads to inflammatory cell infiltration within the field of drug application followed by apoptosis of diseased tissue. This article reviews available data on the use of topical imiquimod for AK. Topical imiquimod is an effective and safe treatment option for AK that produces complete eradication or marked reduction in the number of lesions in most patients. Subclinical lesions also emerge during treatment of the affected skin region ("field treatment"). In addition, there is evidence that topical imiquimod at least partially reverses some of the cellular, molecular, and genetic photocarcinogenic changes that develop in skin damaged by UV light. Recent evidence suggests that many patients who effectively are cleared of AK lesions after topical imiquimod use remain free of lesions for several months to 2 years or develop a minimal number of new AK lesions. PMID- 16315561 TI - Enhancing the care and treatment of skin of color, part 1: The broad scope of pigmentary disorders. AB - Scientific research and technologies related to skin pigmentation and dyschromias, which are often key skin concerns for patients of color, have led to recent developments in skin care and treatment. Differences and similarities between skin of color and white skin and current issues in the treatment of ethnic skin are reviewed. Recent research findings, such as the elucidation of the protease-activated receptor 2 (PAR-2) pathway and its role in pigmentation, and areas for further investigation, such as the pathogenesis of pseudofolliculitis barbae (PFB), also are discussed. Awareness of this information within the wider community of dermatologists, primary healthcare providers, and the media will help to accomplish the objective of stimulating new prospective research. PMID- 16315562 TI - Cold urticaria: a case report and review of the literature. AB - Cold urticaria represents a form of physical urticaria. The disorder is uncommon, and patients with the condition are at risk for systemic reactions and thus must be identified, counseled, and treated accordingly. Diagnosis principally is clinical and is confirmed by the results of cold stimulation tests such as placing an ice cube on the patient's forearm. Treatment primarily consists of preventive counseling, epinephrine autoinjections, and antihistamines. We present the case of a 9-year-old girl with acquired cold urticaria and review the literature. PMID- 16315563 TI - Scurvy masquerading as leukocytoclastic vasculitis: a case report and review of the literature. AB - Scurvy, a disease rarely seen in modern times, results from dietary deficiency of vitamin C and is characterized in adults by hemorrhagic diathesis, hair follicle abnormalities, and osteopenia. We present a 59-year-old man with perifollicular petechiae of the extremities, a painful lower extremity hematoma, and sacral osteopenia, who was repeatedly misdiagnosed with leukocytoclastic vasculitis. The patient's dietary history revealed several months of virtually no vitamin C intake. The patient rapidly improved with vitamin C replacement. We review the biochemical basis and pathophysiology of scurvy, clinical scenarios in which it occurs, clinical signs and radiologic features of the condition, and recommendations for its diagnosis and treatment. PMID- 16315564 TI - Eruptive syringomas: an unusual presentation on the buttocks. AB - Syringomas are benign, eccrine, sweat gland tumors. Eruptive syrin-gomas are rare variants that typically develop on the body's anterior surface. We present the case of a 21-year-old black man with multiple, eruptive, asymptomatic syringomas on his buttocks that had persisted for 3 months. This case is unusual in that the patient's age, race, and sex and the location of the lesions are not considered typical for eruptive syringomas. We also present a review of eruptive syringomas and recent literature. PMID- 16315565 TI - The Taylor Hyperpigmentation Scale: a new visual assessment tool for the evaluation of skin color and pigmentation. AB - The Taylor Hyperpigmentation Scale is a new visual scale developed to provide an inexpensive and convenient method to assess skin color and monitor the improvement of hyperpigmentation following therapy. The tool consists of 15 uniquely colored plastic cards spanning the full range of skin hues and is applicable to individuals with Fitzpatrick skin types I to VI. Each card contains 10 bands of increasingly darker gradations of skin hue that represent progressive levels of hyperpigmentation. This article describes the ongoing development of the Taylor Hyperpigmentation Scale and reports the results of a recent validation study of the use of this newly developed chart in individuals with skin of color. In the study, skin color and an area of hyperpigmentation in 30 subjects of white, African American, Asian, or Hispanic ancestry (approximately 5 from each of the 6 skin types) were evaluated by 10 investigators. The results of the study revealed significant variation among intraindividual and interindividual ratings by investigators of skin hue (P < .0001) and hyperpigmentation (P = .0008); however, most investigators rated the scale as useful and easy to use, and 60% stated they would use it in clinical practice to document the response of hyperpigmentation to therapeutic agents. A heuristic evaluation of the results of this study provided insight into essential considerations for the continued effort to develop a useful and simple scale for assessing skin color and pigmentation. PMID- 16315566 TI - Special considerations when conducting measurements with children. PMID- 16315567 TI - Meta-analysis of the reliability and validity of Part B of the Index of Work Satisfaction across studies. AB - Nurses' job satisfaction is a crucial factor in health care organizations. This study uses meta-analysis for reliability generalization and synthesis of construct validity of Part B of the Index of Work Satisfaction (IWS), a measure of job satisfaction. Meta-analysis was performed including assessments of study quality and descriptive coding of studies. Rater reliability was assessed for all coding and extraction of data. The mean reliability of Part B scores of the IWS based on 14 studies was .78 (df = 13, p < .05). The mean score reliability was .77 for university settings, .73 for community/acute care hospitals, .77 for multi-site studies, and .90 for other settings. For studies rated high and low quality, the mean score reliability was .77 and .83, respectively. Scores on Part B of the IWS correlated -.38 with turnover intent, .60 with organizational commitment, and -.53 with job stress. Scores on Part B of the IWS are reliable for measuring job satisfaction of nurses across samples. Construct validity needs additional testing. PMID- 16315568 TI - Questionnaire development to predict mammography intention among women in southeastern Louisiana. AB - This article summarizes the development and psychometric analysis of the Thoughts About Mammography (TAM) questionnaire to predict mammography intention using the Theory of Planned Behavior (TPB). Key themes, obtained from elicitation interviews (N = 45), were categorized, ranked and extracted for item construction. Initial pilot testing supported test-retest reliability (alpha = .85 to .97), internal consistency (alpha = .67 to .91), and content validity (0.86-1.00). After pilot testing, the TAM was administered to 302 rural women in southeastern (SE) Louisiana. The instrument was internally consistent (alpha = 0.77 to 0.92), construct valid (alpha = .18 to .64), and predicted 24% of the variance of mammography intention. The TAM adequately demonstrated reliability and validity to measure mammography intention in rural southeastern Louisiana women. PMID- 16315569 TI - Development of an instrument measuring family care. AB - This study developed and evaluated an instrument to measure the quality of care given by family members to a patient in the home. The Family Care Measure consisted of 11 items, each of which was rated on a visual analogue scale. Fifteen registered nurses rated the care given by families of 72 eligible patients for whom the nurses had provided nursing care. Estimation of the reliability resulted in a high internal consistency. With deletion of five items from the measure, the internal consistency reliability resulted in a Cronbach's alpha coefficient of .93. The six-item instrument correlated moderately, as predicted, with a scale of family coping (r = .76). With acceptable reliability and evidence of construct validity, this measure may be used in research studies, but further testing to assess its usefulness in clinical situations is recommended. PMID- 16315570 TI - Development and psychometric testing of the Relational Care Scale. AB - The purpose of this article is to report the development and testing of the self report Relational Care Scale (RCS). The RCS was specifically designed to evaluate the relational care that nursing staff provides to residents living in nursing homes. The scale's content validity was assessed with the assistance of experts and related research in the area of nurse-resident interactions, and then the scale was pilot tested with 30 residents of a long-term care (LTC) facility to determine its clarity and stability. The 6-item scale was then administered to a sample of 50 residents in an intervention study and then to 72 residents in a correlational study in two other LTC facilities in Ontario, Canada. Construct validity was demonstrated with positive correlations between RCS scores and a Relationship VAS measure (r = .63, p < .0001) and between RCS scores and an observational Relational Behavior Scale (r = .42, p < .001), which was also developed to assess relational care. Study results demonstrated that the RCS is a reliable and valid instrument that may be used to measure relational care to test interventions focusing on enhancing the relational care staff provides to residents. PMID- 16315572 TI - Take an advocacy role in preventing medical errors in your hospital. PMID- 16315571 TI - Psychometrics of the Laffrey Health Conception Scale for adolescents. AB - The purposes of this methodological study were to factor analyze the Laffrey Health Conception Scale (LHCS) and to assess construct validity of the instrument with early adolescents. The final sample consisted of 230 early adolescents, aged 12 to 14, who responded to instrument packets in classrooms in an urban middle school. Data obtained on the LHCS were subjected to principal components factor analysis with oblique rotation. A two-factor solution was accepted, which is consistent with early adolescents' conceptions of health. Factor I was labeled Wellness and Factor II was labeled Clinical Health. A higher order factor analysis yielded one factor with 26 items, labeled the LHCS for Early Adolescents. The 26-item LHCS had a coefficient alpha of .95. Construct validity was assessed by testing three theoretical propositions, which significantly linked health conception to social support, self-esteem, and positive health practices. The findings indicate that the LHCS is a reliable and valid measure of health conceptions in early adolescents. Results also offer flexibility to researchers interested in testing theory involving the constructs of the definition of health, wellness, and clinical health in early adolescents. PMID- 16315573 TI - Take a role in preventing, reporting medical errors. PMID- 16315574 TI - Community interventions aid self-care, prevention. PMID- 16315575 TI - Administrative support helps keep your LOS low. PMID- 16315577 TI - CPGs may not be best for older patients. PMID- 16315576 TI - ICU patients at risk for preventable errors. PMID- 16315578 TI - Program helps immigrants navigate health system. PMID- 16315579 TI - Achieve case management service excellence. PMID- 16315580 TI - Review of treatment options for lyme borreliosis. AB - Lyme borreliosis (Lyme disease) is the most common tick-borne bacterial infection and the incidence is increasing in parts of Europe and the USA. Prompt antimicrobial therapy using oral agents such as doxycycline or amoxicillin is successful among more than 90% of patients. Inadequate penetration of oral agents into the CNS may result in the development of overt neuroborreliosis. The parenteral agent ceftriaxone is the drug of choice for severe acute and chronic infections, due to good penetration into CSF, convenient single daily dosage regimen and proven high efficacy in clinical trials involving a wide variety of disseminated infections. Regardless of therapeutic agent, there appears to a small minority of patients (<10%) who do not respond; such cases may be due to long-term persistence of borrelial cysts and to misdiagnoses based solely on seropositivity. Several adjunct therapies are available, including hyperbaric oxygen therapy and immune system supplements, but clinical trials have yet to be conducted. PMID- 16315581 TI - Ceftriaxone prophylaxis in abdominal, cardiovascular, thoracic, orthopaedic, neurosurgical and general surgery: a review of practice 1996 - 2003. AB - The microbiology, efficacy and cost-effectiveness of ceftriaxone prophylaxis were compared with those of alternative antimicrobial agents in abdominal, cardiovascular, thoracic, orthopaedic, neurosurgical and general surgical procedures published since 1996. Ceftriaxone was compared with cefazolin +/- metronidazole, cefoxitin, cefuroxime, ceftazidime, cefotaxime, cefepime + metronidazole, penicillins, ticarcillin/clavulanic acid, ampicillin/sulbactam, vancomycin and combined clindamycin/gentamicin. Ceftriaxone, used primarily as 'single shot prophylaxis', was at least as clinically effective if not better than the comparative single- and multiple-dose agents over the broad range of surgical procedures. Furthermore the overall cost of ceftriaxone prophylaxis has often been shown to be markedly less than comparators, despite the relatively high acquisition cost of ceftriaxoe, when factors other than acquisition cost were considered. Advances in surgical techniques, the changes in bacterial ecology in hospitals, the spread of bacterial resistance and the substantial increase in the surgical population at risk suggest that third generation cephalosporins, particularly ceftriaxone, should be taken into consideration for surgical prophylaxis. PMID- 16315582 TI - Predicting the clinical efficacy of generic formulations of ceftriaxone. AB - Time above MIC (T>MIC) is regarded as the best pharmacokinetic/pharmacodynamic (PK/PD) parameter for predicting the clinical efficacy of cephalosporins. The concentration of non-protein-bound proprietary ceftriaxone (Rocephin, Roche) in body fluids exceeds this PK/PD criterion for the treatment of Streptococcus pneumoniae respiratory infections. However, the pharmaceutical quality of several generic products may be inferior to Rocephin. We have calculated the variations in fluid concentrations of 34 generic formulations of ceftriaxone and, by mathematical modelling, the implications for attainment of recommended PK/PD criteria, specifically: Treatment of S. pneumoniae infections based on the time that non-protein-bound ceftriaxone concentration in pleural fluid exceeds the CLSI (NCCLS) breakpoint of 4 mg/L for identification of resistant isolates. Impact upon Monte Carlo simulations in plasma for the treatment of S. pneumoniae infections based on T>MIC for 50% dosing interval. Rocephin exceeded the required PK/PD parameters at the mean and two standard deviation levels in both investigations. In contrast, most generic products failed to achieve required PK/PD levels in both investigations. As a consequence, some generic formulations of ceftriaxone may increase risks of clinical failure and/or emergence of resistant isolates. PMID- 16315583 TI - [Role of obligatory medical insurance in insuring victims of occupational accidents and patients with occupational diseases]. AB - The author described current system insuring victims of occupational accidents and patients with occupational diseases, with participation of all off-budget funds and health service budget, with consideration of work conditions and occupational morbidity in Rostov region. Analysis covered experience and efficiency of Obligatory Medical Insurance System (unique in this country, which participated in insuring occupational victims) in insuring victims of occupational accidents in the region. PMID- 16315584 TI - [Comparative analysis of methods evaluating ergonomic risks]. AB - The authors demonstrated appropriateness of some subjective methods evaluating strain in workers engaged into road building and determined work hardiness categories, considering peculiarities of individual methods. Calculations using empiric equations covered energy consumption during workers' dynamic activities. Empiric equations were proved to point to higher energy consumption, if compared to results of experimental methods. Objective evaluation of work hardiness involved heart rhythm monitoring through a device transforming heart rate into metabolic energy, kcal/min. PMID- 16315585 TI - [Hormonal mechanisms of peripheral neuropathy in vibration disease]. AB - Studies of peripheral limb nerves through stimulation electroneuromyography, evaluation of hormonal levels via radioimmune assay were performed in 59 patients having vibration disease caused by local vibration. Findings are obligatory signs of peripheral neuropathy and hypothyroid state without thyroid disease manifestations. Statistic analysis revealed reliable dependence between degree of motor axons functional deficit and level of serum triiodothyronine and thyroxin decrease--that could point to participation of thyroid hormones in peripheral neuropathy progression in vibration disease. PMID- 16315586 TI - [Studying leucocyte-endothelial mechanisms for diagnosis of vascular disorders in vibration disease]. AB - The article presents pathogenetic mechanisms of microangiopathy development in vibration disease. The authors specified leucocyte-endothelial interactions in vascular dysfunction caused by local vibrations, suggested additional methods to diagnose vascular disorders in vibration disease patients. PMID- 16315587 TI - [Cardiovascular state and efficiency of information processing depend from work load in operators of heat power stations]. AB - Increased working load appeared to cause unfavorable cardiovascular state in operators: dyastolic hypertension stage I, lower than average hemodynamic parameters, lower values of Kerdo index (within negative zone) vs those in lesser occupational load, more marked changes in parameters by the end of working shifts, evening-out of their regular diurnal periodicity. Lower efficiency of information processing was also seen. PMID- 16315588 TI - [Information value of "additional tasks" method to evaluate pilot's work load]. AB - "Additional task" method was used to evaluate pilot's work load in prolonged flight. Calculated through durations of latent periods of motor responses, quantitative criterion of work load is more informative for objective evaluation of pilot's involvement in his piloting functions rather than of other registered parameters. PMID- 16315589 TI - [Physiologic and hygienic characteristics of college teachers work]. AB - First series of studies covered analysis of lecture with registering number of words and movements complementary to them. The series 2 determined occupational activities of college teacher, according to contemporary hygienic classification, as highly intensive work requiring physiologic and managerial correction. PMID- 16315590 TI - [Clinical and electrocardiographic evaluation of cardiovascular system in personnel of chemical metallurgy plant in Siberian Chemical Complex]. PMID- 16315591 TI - [All-Russia scientific and practical conference "Ergonomics of Working Process" (September, 13-14, 2005)]. PMID- 16315592 TI - Progress in studies on the induction and differentiation of embryonic stem cells. AB - Embryonic stem cells (ESCs), which are isolated from the inner cell mass of the blastocyst stage embryo, have the potential to give rise to an entire organism and to generate every body cell type. Much improvement has been made in the field of induction and differentiation of ESCs during the last two years, such as the ESCs differentiation into germ cells (2003) and the cloning of human ESCs (2004), both of which were chosen respectively as one of the top ten achievements evaluated by academic journals. Great attention was also paid to the research of the new genes which could maintain ESCs in the undifferentiated state and the research of the induction and differentiation of ESCs. PMID- 16315593 TI - Alien DNA introgression and wheat DNA rearrangements in a stable wheat line derived from the early generation of distant hybridization. AB - Polyploidy has been found to be common in plants. Bread or common wheat (Triticum aestivum L., 2n=42) is a good example of allopolyploid made up of three diploid genomes A, B and D. In recent years, by the study of mimicking the origination of common wheat, it was found that changes of DNA sequence and gene expression occurred at the early stages of artificial allohexaploid between tetraploid wheat and Aegilops tauschii, which was probably favorable to genetic diploidization of new synthetic hexaploid wheat. Common wheat 99L2 is a new line stable in genetic, which was derived from the early self-pollinated generation of wide hybrids between common wheat and rye. In this study, it was found that at least two rye DNA segments had been introgressed into 99L2. This result suggested that a mechanism of alien DNA introgression may exist, which was different from the traditional mechanism of chromosome pairing and DNA recombination between wheat and alien species. Meanwhile, during the introgression process of alien rye DNA segments, the changes in DNA sequences of wheat itself occurred. PMID- 16315594 TI - Exogenous ethylene influences flower opening of cut roses (Rosa hybrida) by regulating the genes encoding ethylene biosynthesis enzymes. AB - The purpose of this paper is to investigate the differential responses of flower opening to ethylene in two cut rose cultivars, 'Samantha', whose opening process is promoted, and 'Kardinal', whose opening process is inhibited by ethylene. Ethylene production and 1-aminocyclopropane-1-carboxylate (ACC) synthase and oxidase activities were determined first. After ethylene treatment, ethylene production, ACC synthase (ACS) and ACC oxidase (ACO) activities in petals increased and peaked at the earlier stage (stage 3) in 'Samantha', and they were much more dramatically enhanced and peaked at the later stage (stage 4) in 'Kardinal' than control during vasing. cDNA fragments of three Rh-ACSs and one Rh ACO genes were cloned and designated as Rh-ACS1, Rh-ACS2, Rh-ACS3 and Rh-ACO1 respectively. Northern blotting analysis revealed that, among three genes of ACS, ethylene-in- duced expression patterns of Rh-ACS3 gene corresponded to ACS activity and ethylene production in both cultivars. A more dramatic accumulation of Rh-ACS3 mRNA was induced by ethylene in 'Kardinal' than that of 'Samantha'. As an ethylene action inhibitor, STS at concentration of 0.2 mmol/L generally inhibited the expression of Rh-ACSs and Rh-ACO in both cultivars, although it induced the expression of Rh-ACS3 transiently in 'Kardinal'. Our results suggests that 'Kardinal' is more sensitive to ethylene than 'Samantha'; and the changes of Rh-ACS3 expression caused by ethylene might be related to the acceleration of flower opening in 'Samantha' and the inhibition in 'Kardinal'. Additional results indicated that three Rh-ACSs genes were differentially associated with flower opening and senescence as well as wounding PMID- 16315595 TI - Isolation and annotation of 10828 putative full length cDNAs from indica rice. AB - We reported the isolation and identification of 10828 putative full-length cDNAs (FL-cDNA) from an indica rice cultivar, Minghui 63, with the long-term goal to isolate all full-length cDNAs from indica genome. Comparison with the databases showed that 780 of them are new rice cDNAs with no match in japonica cDNA database. Totally, 9078 of the FL-cDNAs contained predicted ORFs matching with japonica FL-cDNAs and 6543 could find homologous proteins with complete ORFs. 53% of the matched FL-cDNAs isolated in this study had longer 5'UTR than japonica FL cDNAs. In silico mapping showed that 9776 (90.28%) of the FL-cDNAs had matched genomic sequences in the japonica genome and 10046 (92.78%) had matched genomic sequences in the indica genome. The average nucleotide sequence identity between the two subspecies is 99.2%. A majority of FL-cDNAs (90%) could be classified with GO (gene ontology) terms based on homology proteins. More than 60% of the new cDNAs isolated in this study had no homology to the known proteins. This set of FL-cDNAs should be useful for functional genomics and proteomics studies. PMID- 16315596 TI - Functional analysis of a reproductive organ predominant expressing promoter in cotton plants. AB - Transgenic Bt insect-resistant cotton plants have high insect resistance in the early stage of development, but relatively low resistance in the late stage. Substituting a reproductive organ-specific promoter for the CaMV35S promoter presently being used could be an ideal solution. For the first time, the promoter sequence of ADP-ribosylation factor 1 (arf1) gene was isolated from Gossypium hirsutumY18 by means of inverse PCR. The sequencing result discovered the unique structure of the arf1 promoter, including four promoter-specific elements, the initiator, TATA box, CAAT box and GC box, and also an intron in 5'-untranslation region. Four plant expression vectors were constructed for functional analysis of the promoter. Based on the pBl121 plant expression vector, four truncated arf1 promoters took the place of the CaMV35S promoter. These vectors were different only in their promoter regions. They were introduced into cotton plants via pollen tube pathway. Histochemical GUS staining and fluorescence quantitative analyses were performed to examine the expression patterns of the GUS gene driven by the 4 arf1 truncated promoters in transgenic cotton plants respectively. The results showed that the arf1 promoter was a typical reproductive organ-specific promoter. Hopefully, the arf1 promoter can be a regulatory element for designing cotton reproductive organs with desired characteristics. PMID- 16315597 TI - Identification of prior candidate genes for Sclerotinia local resistance in Brassica napus using Arabidopsis cDNA microarray and Brassica-Arabidopsis comparative mapping. AB - Arabidopsis cDNA arrays were used to screen the local-defense-associated genes in oilseed rape (Brassica napus L.) at the challenge of Sclerotinia sclerotiorum. 61 genes with two-fold expression changes were screened out from the local tissue around the necrosis. Among them, 36 unique genes were up-regulated and 25 unique genes were down-regulated. RT-PCR and Northern blot results were consistent with the array results, suggesting Arabidopsis arrays were useful for transcriptional profiling of B. napus genes. Some of these genes were located in the interval of some QTLs for Sclerotinia resistance in B. napus by Brassica-Arabidopsis comparative mapping. These genes may have priority to be pursued for more intensive research. PMID- 16315598 TI - Differentiation of embryonic stem cells into corneal epithelium. AB - Our project was to determine whether embryonic stem (ES) cells could be induced to differentiate into corneal epithelia by superficial corneoscleral limbal stroma. To achieve this goal, ES-GFP cell line D3 was pre-induced by retinoic acid (RA). The pre-induced cells were seeded on deepithelialized superficial corneoscleral slices (SCSS) to form a monolayer, and divided into three groups. Group 1 was cultured and passaged in vitro for direct detection. Group 2 was exposed to air-liquid interfaces for 10 days and implanted into the subcutaneous layer of nude mice for 2 weeks for further induction in vivo. Group 3 was cultured in vitro without any inducing factors for control. There were no teratomas found in nude mice which were implanted with differentiated ES cells after two weeks. The differentiated cells showed an appearance of epithelia both in vitro and in vivo. Expression of CK3, P63 and PCNA was detected by immunohistochemical staining in the differentiated cells in group 1 and 2. Microvillis and zonula occludens were observed on the surface of the differentiated cells under an electron microscope. In the control group, ES cells differentiated freely without any inducing factors. Most cells were shed and formed a neuronal dendrite-like structure, and a minority of cells appeared polymorphic. These results demonstrate that ES cells can differentiate into corneal epithelia on the surface of SCSS under the controlled condition. Differentiated ES cells could be used as epithelial seeding cells for the reconstruction of ocular surface and corneal tissue engineering in the future. PMID- 16315599 TI - Research of blastocyte-like structure in chicken. AB - The chicken embryo is a classic model used to investigate embryonic development, gene expression, and tissue differentiation, and is also an important research tool in studying the animal functional genomics. The whole blastoderms of fresh unincubated eggs from White Leghorn chickens were collected with a paper ring, mechanically broken into small pieces and cultured in medium. Then the small pieces would develop into blastocyte-like structures (BLS), which could be facilitated by an addition of fetal bovine serum (FBS) to the primary culture and their diameter was nearly doubled from 12 to 24 h. The additional yolk had no positive effect on the development in the first 12 h but encouraged the BLSs attaching and inner cells differentiating instead in 24 h. The inner cells of the BLS showing a high alkaline-phosphatase activity similar to those in mouse embryonic stem (ES) cells and also expressing a large amount of the specific stage embryonic antigen-1 (SSEA-1) on the surface, which was known to be the characteristic of non-differentiated mouse and avian ES cells, could finally differentiate into nerve-like cells, fibroblast cells and so on in the medium. Leukemia inhibitory factor (LIF) facilitated the cells' proliferation and prevented differentiation in the suspended culture of the BLSs. So we drew the conclusion that the BLS obtained from broken blastoderm can be used to amplify avian ES cells so as to initiate a new method of producing transgenic chickens. PMID- 16315600 TI - Effect of the infectious laryngotracheitis virus (ILTV) glycoprotein G on virus attachment, penetration, growth curve and direct cell-to-cell spread. AB - The secreted alphaherpesvirus glycoprotein G (gG) works differently from other proteins. Analysis of the role of ILTV gG in virus attachment, penetration, direct cell-to-cell spread (CTCS) and the growth curve showed that gG or its antibody had no effect on ILTV attachment and penetration and that the gG antibody reduced the virus plaque size and the one-step growth curve on chicken embryo liver (CEL) cells, but gG did not affect the virus plaque size or the one step growth curve on CEL cells. Laser scanning confocal microscopy (LSCM) detection showed that ILTV gG is located in the perinuclear region and the membrane of the CEL cells. These results suggested that ILTV gG might contribute to direct cell-to-cell transmission. PMID- 16315601 TI - Effects of epoxyeicosatrienoic acids on levels of eNOS phosphorylation and relevant signaling transduction pathways involved. AB - Endothelial nitric oxide synthase (eNOS) is a key enzyme responsible for the regulation of vascular homeostasis. Many humor factors and mechanical forces can affect eNOS activity via phosphorylation modification but the mechanisms involved vary with stimuli applied. We have demonstrated that cytochrome P450 (CYP) epoxygenase-dependent metabolites of arachidonic acid, epoxyeicosatrienoic acids (EETs), can robustly up-regulate eNOS expression and its activity, however the relevant signaling pathways responsible for activity regulation are not well known. In this study, we explored the role of PI3 kinase (PI3K)/protein kinase B (Akt) signaling pathway in eNOS expression and its phosphorylation in response to EETs via direct addition of EETs into cultured bovine aorta endothelial cells (BAECs) and recombinant adeno-associated virus-mediated transfection of CYP epoxygenase genes CYPF87V and CYP2C11 to produce endogenous EETs followed by co treatment with PI3K or Akt inhibitor. Results show that both exogenous and endogenous EETs could remarkably enhance eNOS expression and its phosphorylation at Ser1179 and Thr497 residues; PI3K inhibitor LY294002 could inhibit EETs induced increase in eNOS-Ser(P)1179 but had no effect on the change of eNOS Thr(P)497, while Akt inhibitor could attenuate the increase in phosphor-eNOS at both residues; both of the two inhibitors could block EETs-enhanced eNOS expression. These results lead to conclusions: (i) EETs-mediated regulation of eNOS activity may be related with the changes of phosphorylation level at eNOS Ser1179 via P13K/Akt and eNOS-Thr497 via Akt; (ii) PI3K/Akt signaling pathway is involved in the up-regulation of eNOS expression by EETs. PMID- 16315602 TI - Effects of ELF magnetic fields on protein expression profile of human breast cancer cell MCF7. AB - Extremely Low Frequency Magnetic Fields (ELF MF) has been considered as a "possible human carcinogen" by International Agency for Research on Cancer (IARC) while credible mechanisms of its carcinogenicity remain unknown. In this study, a proteomics approach was employed to investigate the changes of protein expression profile induced by ELF MF in human breast cancer cell line MCF7, in order to determine ELF MF-responsive proteins. MCF7 cells were exposed to 50 Hz, 0.4 mT ELF MF for 24 h and the changes of protein profile were examined using two dimensional electrophoresis. Up to 6 spots have been statistically significantly altered (their expression levels were changed at least 5 fold up or down) compared with sham-exposed group. 19 ones were only detected in exposure group while 19 ones were missing. Three proteins were identified by LC-IT Tandem MS as RNA binding protein regulatory subunit, Proteasome subunit beta type 7 precursor and Translationally Controlled Tumor Protein. Our finding showed that 50 Hz, 0.4 mT ELF MF alternates the protein profile of MCF7 cell and may affect many physiological functions of normal cell and 2-DE coupled with MS is a promising approach to elucidating cellular effects of electromagnetic fields. PMID- 16315603 TI - Induction of xenogeneic islet transplantation tolerance by simultaneously blocking CD28-B7 and OX40-OX40L co-stimulatory pathways. AB - It has been demonstrated that prolonged graft survival can be achieved through inhibiting the activation of T cells, and addition of soluble CTLA4Ig and OX40Ig proteins to mixed lymphocyte reactions can effectively inhibit T cell proliferation. To explore the potential of this type of treatment in xenotransplantation, we infected streptozotocin-induced diabetic BalB/c mice (H 2d) (200 mg/kg, IV) with 5 x 10(8) pfu AdCTLA4Ig-IRES-OX40Ig on day 1 before islets transplantation through the tail vein. The results showed that this treatment prolonged the islet xenografts survival significantly. The reaction to exogenous glucose stimulation was normal and the cytokine secretion of the type Th1 cells was inhibited. The AdCTLA4Ig-IRES-OX40Ig-mediated treatment effectively induced the T cells into anergy and the Th1/Th2 cells into deviation. These results strongly supported the therapeutic potential of blockade of costimulation by AdCTLA4Ig-IRES-OX40Ig genes transfer in inducing the organ transplantation tolerance. PMID- 16315604 TI - Tyroserleutide tripeptide affects calcium homeostasis of human hepatocarcinoma BEL-7402 cells. AB - This study aimed to observe the effects of tyroserleutide (tyrosyl-seryl-leucine, YSL) on the growth of human hepatocarcinoma BEL-7402 that was transplanted into nude mice, and explore its anti-tumor mechanism preliminarily. YSL, at doses of 80 microg x kg(-1) x d(-1), 160 microg x kg(-1) x d(-1) and 320 microg x kg(-1) x d(-1) significantly inhibited the growth of the human hepatocarcinoma BEL-7402 tumor in nude mice, producing inhibition of 21.66%, 41.34%, and 34.78%, respectively. Ultra structure of BEL-7402 tumor in nude mice showed that YSL could induce tumor cells apoptosis and necrosis, cell organelle mitochondria and endoplasmic reticulum damage, and calcium overload. By confocal laser scanning microscopy and flow cytometry, we found that 10 microg/mL YSL rapidly induced an increase of the concentration of cytoplasmic free calcium in BEL-7402 cells in vitro, and maintained high concentrations of cytoplasmic free calcium for 1 h. Then the calcium concentration began to decrease after 2 h, and was lower than that of the control group at 4 h and 24 h (p < 0.05). YSL also decreased the mitochondrial transmembrane potential of BEL-7402 cells in vitro, but had no effect on the calcium homeostasis or mitochondrial transmembrane potential of Chang liver hepatocytes. So affecting calcium homeostasis, then inducing apoptosis and necrosis may be a mechanism by which YSL inhibits the tumor growth in animal model. PMID- 16315605 TI - New strategies for CNS repair. PMID- 16315606 TI - Heterogeneity of multiple sclerosis: implications for therapy targeting regeneration. PMID- 16315607 TI - The neuroprotective effect of inflammation: implications for the therapy of multiple sclerosis. PMID- 16315608 TI - Fibroblast growth factors in oligodendrocyte physiology and myelin repair. PMID- 16315609 TI - White matter progenitor cells reside in an oligodendrogenic niche. PMID- 16315610 TI - At the interface of the immune system and the nervous system: how neuroinflammation modulates the fate of neural progenitors in vivo. AB - Neural stem and progenitor cells express a variety of receptors that enable them to sense and react to signals emanating from physiological and pathophysiological conditions in the brain as well as elsewhere in the body. Many of these receptors and were first described in investigations of the immune system, particularly with respect to hematopoietic stem cells. This emerging view of neurobiology has two major implications. First, many phenomena known from the hematopoietic system may actually be generalizable to stem cells from many organ systems, reflecting the cells' progenitor-mediated regenerative potential. Second, regenerative interfaces may exist between diverse organ systems; populations of cells of neuroectodermal and hematopoietic origin may interact to play a crucial role in normal brain physiology, pathology, and repair. An understanding of the origins of signals and the neural progenitors' responses might lead to the development of effective therapeutic strategies to counterbalance acute and chronic neurodegenerative processes. Such strategies may include modifying and modulating cells with regenerative potential in subtle ways. For example, stem cells might be able to detect pathology-associated signals and be used as "interpreters" to mediate drug and other therapeutic interventions. This review has focused on the role of inflammation in brain repair. We propose that resident astroglia and blood-born cells both contribute to an inflammatory signature that is unique to each kind of neuronal degeneration or injury. These cells play a key role in coordinating the neural progenitor cell response to brain injury by exerting direct and indirect environmentally mediated influence on neural progenitor cells. We suggest that investigations of the neural progenitor-immunologic interface will provide valuable data related to the mechanisms by which endogenous and exogenous neural progenitor cells react to brain pathology, ultimately aiding in the design of more effective therapeutic applications of stem cell biology. Such improvements will include: (1) ascertaining the proper timing for implanting exogenous neural progenitor cells in relation to the administration of anti-inflammatory agents; (2) identifying what types of molecules might be administered during injury to enhance the mobilization and differentiation of endogenous and exogenous neural progenitor cells while also inhibiting the detrimental aspects of the inflammatory reaction; (3) divining clues as to which molecules may be required to change the lesioned environment in order to invite the homing of reparative neural progenitor cells. PMID- 16315611 TI - Remyelination and restoration of axonal function by glial cell transplantation. PMID- 16315612 TI - Gene and stem cell therapy for autoimmune demyelination. PMID- 16315613 TI - Novel gene therapeutic strategies for neurodegenerative diseases. AB - The convergent pathobiologic model of Parkinson's disease stipulates that disparate insults initiate a disease process that obligately share a common pathway leading to cell death. A combinatorial treatment which targets various steps in this pathway is likely to be the most successful therapeutic strategy. As advances are made in the field of neuroimaging and pharmacogenomics, early detection of sporadic PD will become a reality. Early intervention will likely spare more dopaminergic neurons and extend the quality of life for the patient. Continued advancements in the fields of pharmacology, neurosurgery, and gene therapy will strengthen the armamentarium available for the treatment of PD patients. PMID- 16315614 TI - Measuring injury and repair of myelin and neurons in multiple sclerosis. AB - Neuroprotection in MS needs to be considered in the context of several pathological processes: limitation of acute inflammatory injury to myelin and axons, remyelination, survival of demyelinated axons, and limitation of more diffuse, nonlesional pathology that affects myelin and axons. Advanced MRI techniques are capable of reporting on all of these different pathological features of MS and will be an important aspect of the assessment of neuroprotection strategies in MS, when these become available. PMID- 16315615 TI - Medical education in Malaysia. PMID- 16315616 TI - Medical education today: globalising with quality. AB - With globalization education has become a tradable service governed by the rules and regulations of GATS and worth trillions of dollars. International standards are rapidly being developed to facilitate cross border supply of services. In medical education, the WFME has produced International Guidelines on Quality in Medical Education which has a regional equivalent in the WHO Western Pacific Region, and the IIME has defined the minimum essential requirements of standards in medical education in seven core competences. Malaysia, having an explicit policy of making education a sector for revenue generation, has put in place regulatory frameworks and incentives to make the country a centre of educational excellence. Within the ambit of this national aspiration, medical education has grown phenomenally in the last decade. Standards and procedures for accreditation of medical schools in line with the world standards have been developed and implemented and policies are enforced to facilitate compliance to the standards. The ultimate goal is for medical schools to be self-accredited. In striving towards self-accreditation medical schools should be innovative in making changes in the three requirements of medical education. These are the intellectual and social imperatives and strategies for effective implementation. PMID- 16315617 TI - Accreditation of medical schools in Ireland: standards and procedures. AB - The Irish Medical Council has undertaken accreditation inspections of Irish medical schools on a regular basis since 1996. This document is a summary of the accreditation standards, a guide to the process for those involved and an overview of the complexity of the many elements involved in educating a doctor. It should be read in conjunction with previous Medical Council publications on medical education. It also provides the basis for the Evaluation System for Visitors 2003. The Medial Council's prime role is the protection of the public interest in relation to the practice of medicine. The Medical Council scrutinises medical schools. It has an important advocacy role with government, with the universities which operate medical schools and with the professionals involved to improve the standards and delivery of medical education. PMID- 16315618 TI - Accreditation of medical training in Australia and New Zealand. AB - The Australian Medical Council (AMC) accredits both Australian and New Zealand (NZ) medical courses and also college specialist training programmes. The common accreditation process allows mutual recognition of basic medical training and vocational training between Australia and New Zealand. The ultimate purpose of accreditation assure stakeholders including medical registration boards, health departments, students/trainees and the general community of the quality of the programs and the competence of those completing such training. AMC revised its own accreditation guidelines using the WFME standards as the model around which the new AMC standards were developed. The College Accreditation Process is similar to and builds on AMC experience in the medical school accreditation process. In conclusion, AMC accreditation has been successful in improving medical education in Australia and New Zealand and has been able to do so without the imposition of any exclusive educational model or philosophy. PMID- 16315619 TI - Future needs and wants in medical education--the UPM experience. AB - Medical schools have existed in Malaysia for a very long time. The majority of practicing doctors has trained locally. From the early nineteen sixties when the first medical school was established to the current 7 public medical faculties, the standards of local trained doctors have always been exemplary. Now with more need for doctors to serve the country and the mushrooming of medical schools, the question of needs and wants of future medical schools and medical education need to be addressed. In this paper I will try to highlight what we in Universiti Putra Malaysia have been working on to improve the Medical and Health Sciences faculty to achieve greater heights to reach the future in the shortest possible time. PMID- 16315620 TI - The role of the Malaysian Medical Council in medical education. AB - The Malaysian Medical Council (MMC) operates under the Medical Act of 1971, which defines its core functions related to (a) the registration and practice of medical practitioners (b) the period of compulsory service (c) provisions to be enacted for purposes of (a) and (b). In the early years the MMC used the list of recognised colleges or Universities that appeared in the list of degrees recognised by the General Medical Council of United Kingdom (GMC). Over the years the MMC has undertaken the role of granting recognition to other medical schools in the country and overseas, and added the name of these schools to the existing register of recognised medical degrees in the second schedule of the Act. For the purpose of recognition of medical schools the MMC endorsed a guideline on standards and procedures on accreditation developed in 1996, which was later realigned with international and regional guidelines, in 2000 and 2001. It is recommended that the MMC establishes an active functional 'Education Committee' and that the role of MMC in medical education should be clearly and explicitly stated in the Act. An amendment to the Act would require the MMC to be responsible not only for undergraduate medical education but medical education in its entire phase. PMID- 16315621 TI - Proceedings in a disciplinary action at the Malaysian Medical Council. AB - Members of the medical profession are expected to be well aware and abide by the revised code of ethics adopted by the Malaysian Medical Council on 9th December 1986. Under the Act Council may, in the exercise of its disciplinary jurisdiction, impose punishments related to misconduct or malpractices. When a complaint or information is made against any practitioner, the President shall forward such complaint to the Chairman of the Preliminary Investigation Committee. The procedure of the disciplinary inquiry is not exactly like those in the court of law but the same principle of justice is adhered to and all evidence used to make a decision must only be those that are admissible in accordance with the rule of evidence. PMID- 16315622 TI - Accreditation of medical schools: the question of purpose and outcomes. AB - Accreditation is a process by which official accrediting bodies evaluate institutions using a set of criteria and standards, following established procedures, to ensure a high quality of education needed to produce highly competent graduates. Additional objectives include (1) ensuring quality institutional functioning, (2) strengthening capabilities of educational institutions for service to the nation and (3) improving public confidence in medical schools. The accreditation process provides an opportunity for the institution to critically reflect upon all the aspects of its programme and the level of compliance or attainment of the requirements. The self-evaluation exercise, which identifies strengths and weaknesses, is perceived as formative. It is envisaged that eventually institutions will adopt a learning culture for curriculum development, implementation, monitoring and matching the outcomes. In conclusion, periodic accreditation activities can act as a "monitoring" system to ensure that the quality of medical education is maintained according to established standards. PMID- 16315623 TI - Many roads lead to Rome--is that true of the teaching and learning of pharmacology? Lessons from three medical schools in Malaysia. AB - In Malaysia many new medical schools (both public and private) have been set up in the last 12 years. As a result of global changes and local adjustments made in medical training, cross-breeds of different medical curricula have produced a wide spectrum of teaching-learning methods in these medical schools. In this paper, we have selected three medical schools--two public (Universiti Malaya and Universiti Putra Malaysia) and one private (International Medical University) to illustrate different approaches in the teaching-learning of pharmacology that exist in Malaysia. How do these different teaching-learning approaches affect the students' interest and ability to "master" pharmacology and in turn to develop a good prescribing practice? PMID- 16315624 TI - The use of OSCE (Objective Structured Clinical Examination) as an assessment tool -initial experiences at the University of Malaya. AB - Formative and summative student assessment has always been of concern to medical teachers, and this is especially important at the level of graduating doctors. The effectiveness and comprehensiveness of the clinical training provided is tested with the use of clinical cases, either with real patients who have genuine medical conditions, or with the use of standardised patients who are trained to simulate accurately actual patients. The Objective Structured Clinical Examination (OSCE) is one method of assessing the adequacy of clinical skills of medical students, and their level of competence. It can be used to test a variety of skills such as history taking (communication and interpersonal skills) and performing aspects of physical examination, undertaking emergency procedures, and interpreting investigational data. It can also be used to ensure an adequate depth and breadth of coverage of clinical skills expected of a graduating doctor. PMID- 16315625 TI - The Community Follow-up Project (CFUP). AB - The Community Follow-up Project (CFUP) is a project where medical students choose a hospital in-ward patient during their clinical ward-based attachments and follow-up this patient's progress after discharge from the hospital. The students do a series of home visits and also accompany their patients for some of their follow-ups at the hospital, government clinics, general practitioners' clinics and even to the palliative care or social welfare centres. The students assess the physical, psychological and social impact of the illness on the patient, family and community. By following their patients from the time their patients were in the hospital and back to their homes and community, the students are able to understand in depth the problems faced by patients, the importance of communication skills in educating patients on their illness and the importance of good communication between primary, secondary and tertiary care. PMID- 16315626 TI - The medical school curriculum at University Malaysia Sabah. AB - The integrated curriculum at the newly established medical school at University Malaysia Sabah is examined from aspects of the objectives of the medical training in achieving development of the required skills and knowledge as well as personal and professional development. The teaching is spread over five years with an emphasis on basic medical sciences in the first two years although the students are exposed to clinical skills right from the onset. A gradual transition to emphasis on the acquisition of clinical skills occurs from the third year onwards. However, community medicine and professional development are incorporated into the programme from the first year and are carried over to the final year. Although there are examinations to be passed in all the courses taught every semester, with a Cumulative Grade Point Average (CGPA) of 3.0 (65 percentile score) and the candidate has to pass all the examinations in that year to clear a particular year, two professional examinations are administered, one at the end of the Third Year (end of the Phase I of the Medical Programme) and another at the end of the Fifth or Final year (end of the Phase II of the Medical Programme). Programmes for Postings, Shadow House Officers (SHOP) and Population Health are also incorporated into the curriculum. Delivery of the courses involve Lectures, Self-Learning Packages (SLP), Small Group Discussions (SGD), Seminars, Debates, Dramas, Video clips, Special Study Modules (SSM), Computer-Aided Instruction (CAI), Problem-based Learning (PBL), Problem-solving Sessions (PSS) and Clinical Skills Learning (CSL). The examination involves elements of continuous assessment and final end of semester or end of phases I and II Professional Examinations. Practical may involve Objective Structured Practical Examinations (OSPE) and/or Objective Structured Clinical Examinations (OSCE). They may also involve viva voce and/or short and long case presentations and assessment of log book entries. PMID- 16315627 TI - Management studies in medical education. AB - In 1977, the World Health Assembly (WHA) set the social target--the "Health For All" goal and in 1995, urged member states to "re-orientate medical education and medical practice for "Health For All" (resolution WHA 48.8). This led to World Health Organisation to enunciate the "5-star doctor" needing skills in healthcare management, quality assurance and health economics. The Faculty of Medicine, University of Malaya introduced the New Integrated Curriculum (NIC) in 1995. The objective was aimed at producing a competent doctor with a holistic approach to the practice of medicine. This was to be achieved by having 3 strands of studies i.e. The Scientific Basis of Medicine (SBM), the Doctor, Patient, Health and Society (DPHS), and Personal and Professional Development (PPD) over the 5-year programme, split into 3 phases. Elements of the "5-star doctor" were introduced in strand 2--DPHS and strand 3--PPD. Management studies were introduced in the Personal and Professional Development (PPD) strand. This led to an instructional module--"Principles of Management in Health Care Services (PMGT)" comprising of the Management of Self, Resources and People and incorporating a three week field programme. Evaluation is undertaken at the end of the phase IIIA of the studies. This NIC approach will be able to produce a "5-star doctor", a team player, leader, communicator and an effective manager. PMID- 16315628 TI - Skills assessment for undergraduate orthopaedic education. PMID- 16315629 TI - Recognition of higher medical institutions in Russia. AB - The Russian Federation of higher medical institutions get State accreditation, if their activity conforms to criteria determined by the Ministry of Public Health and the Ministry of Education of the Russian Federation. Kursk State Medical University (KSMU) has a confirmed to requirement of accreditation by the Russian Federation, to conduct annually training of about 5000 students at 12 faculties. KSMU carries out pre-medical undergraduate and postgraduate training in the specialty "Doctor of medicine". For the first time in Russia KSMU was allowed to conduct a 6-year medical training with the use of English as an intermediary language by the Ministry of Public Health and the Ministry of Education. In this relation programmes of training teachers for conducting instruction with the use of an intermediary language (English) and training students Russian with the level necessary for free communication with Russian patients and staff of the clinics have been developed and realized. PMID- 16315630 TI - The Medical Council of India and its revolutionary, way of allowing Indian medical graduates from foreign universities to practice in India. AB - This paper attempts to sensitize the participants to understand the benefits of looking at the regulations of accreditation for medical courses in the neighbouring nearby countries. Deregulation of Medical Education like what they have done will bring enormous revenue benefits for the existing assets like the airports, hospitals, hotels resorts and the communication infrastructure of Malaysia. PMID- 16315631 TI - Recognition of medical universities in Ukraine. AB - The paper presents a general characteristic of the organization of teaching of medicine to foreign students at the Crimean State Medical University. The Crimea State Medical University is a state higher educational establishment having the 4th highest level of state accreditation. The University prepares junior specialists and bachelors in specialties such as nursing, orthopedic dentistry, pharmacy and doctors in general medicine, dentistry and clinical pharmacy. At present there are 1,500 foreign students from 34 countries studying at the university, with more than half are students from Malaysia. The quality of education at CSMU is evaluated by the State accreditation commission an authorized central executive power in the field of education and science. Textbooks and manuals written in English, which have passed expert evaluation at a state level and approved by the Ministry of Public Health of Ukraine, was given permission to be used by all medical schools of the country. PMID- 16315632 TI - [Pros and cons of the controlled clinical trials registry]. PMID- 16315633 TI - [Immunological characteristic of children vertically infected with HIV: a case control study]. AB - HIV infection in children causes a serious immunodeficiency with special characteristics that distinguish it from the adult, causing a global immune deficit. This is a case-control study between Cuban paediatric patients infected with HIV by vertical transmission and a control group of supposedly healthy children. Both groups were characterized from the clinical point of view and markers were used for evaluating the immunologic and virologycal state. Clinically 75% of patients present a pattern of precocious progression; from the total, only two stayed asymptomatic. All HIV-infected children receive antirretroviral treatment and three of them present values of viral load bigger than 100,000 cp/mL. The immune alterations found in the HIV-infected children compared with healthy children were: a cellular immune depletion with diminished counts of lymphocyte subsets T CD4+, CD16+/CD56+ and CD19+, an increase in subsets of CD3+, CD8+, CD8+/CD38+, CD3+/ CD95+ and a hypergammaglobulinemia due to prevalence of immunoglobulin gamma IgG (p < 0.05). On the other hand, there were not significantly differences in the serum levels of both C3 and C4, as well as in the haemolytic activity of the classic and alternate activation pathways of the complement system. This finding allowed better attention and treatment of paediatric HIV patients. PMID- 16315634 TI - [Relationship between morphological diagnosis of NASH (non-alcoholic steatohepatitis) and liver function tests in a group of patients with morbid obesity]. AB - The non-alcoholic steato-hepatitis (NASH) is a common disorder in obese, type 2 diabetics, female and patients with dislipidaemia. Hepatic biochemical test are abnormal. Despite the lack of its own morphological characteristics, NASH can be differentiated from other pathologies, the gold standard for diagnosis is liver biopsy. MATERIAL AND METHODS: We designed a retrolective, comparative, observational and cross-sectional study. Thirty-five obese subjects (11 men and 24 women) who underwent to bariatric surgery and liuer biopsy were included. Data were taken from clinical files, such as anthropometric and biochemical test. Those who had clinical history of related alcohol ingestion or liver damage related to drugs were excluded. A experimented pathologist classified the biopsies according to Brunt classification. Liver slides were classified according to 1) presence of NASH; 2) Inflammation and 3) Fibrosis. Differences between groups were analysed by ANOVA and Spearman correlation. RESULTS: We found differences between women (w) and men (m) for height (m: 1.71 +/- 0.9 vs. w:1.60 +/- 0.09m); weight (m:172.5 +/- 39.1 vs. w:126.9 +/- 24.1kg) and BMI (m:58.2 +/- 9.8 vs. w:49.8 +/- 9) , but not for NASH frequency. Nevertheless subjects with NASH (n = 29, 82.8%o) were older than those without NASH (38.3 +/- 9.6 vs. 29.5 +/- 5.2) and had higher aminotrasferases serum levels (AST: 33.1 +/- 19.2 vs. 23.7 +/- 6.3 ULIL; ALT: 36.5 +/- 19.8 vs. 20.3 +/- 7.6ULIL). NASH pa- tients and those with higher grade of histological inflammation had increment of transaminases and albumin levels. Fibrosis showed correlation only with AST (p = 0.020) and ALT (p = 0.002). CONCLUSION: The NASH frequency in patients who underwent to bariatric surgery for weight reduction is very high (82.8%) and exists correlation among liver test and histological findings but not with clinical because the clinical diagnosis is complicated. PMID- 16315635 TI - [Characteristics of Mexican asthmatic subjects attending an outpatient service]. AB - BACKGROUND: Despite the high prevalence of asthma in Mexico, studies describing general characteristics of these patients are scarce. OBJECTIVE: To analyze the sociodemographic, clinical and respiratory functional profile of asthmatic patients attending for the first time the outpatient service of a tertiary-level hospital, as well as to investigate treatments already received and the abandonment rate. METHODS: Adolescent and adult asthmatic subjects seen from February 2000 to November 2002 in the outpatient service of the National Institute of Respiratory Diseases, in Mexico City, were prospectively studied. Sociodemographic information was gathered and clinical and spirometric evaluations were carried out. The severity of the disease was classified according to international criteria. RESULTS: A total of 1403 patients between 12 and 82 years of age, with predominance of the female gender, were included in the analysis. An 88.7% of patients had the mildest forms of the disease, 10% had moderate asthma, and 1.3% had severe asthma. While 21.4% had never received anti asthma treatment, about one third of the remaining patients had received corticosteroids, independently of the asthma severity, indicating an over treatment of the intermittent form of the disease and an under-treatment of persistent forms. A high abandonment rate (65.7%) was observed. A positive bronchodilator test was observed in 30.3% of patients with intermittent asthma, though this proportion increased to about 67% in those with moderate and severe asthma. CONCLUSION: Mildest forms of the disease predominate among asthmatic patients attending for the first time an outpatient service, with a still very high frequency of inadequate use of corticosteroids. PMID- 16315636 TI - [Decrease of folic acid and cognitive alterations in patients with epilepsy treated with phenytoin or carbamazepine, pilot study]. AB - INTRODUCTION: Phenytoin and carbamazepine were the antiepileptic drugs most frequently used in Mexico and throughout the world. Epileptic patients who take these drugs have a variety of collateral effects including the decrease of folates plasmatic level. Low seric folic acid concentration has been associated with a decline in cognitive functions. The administration of a combined treatment with folic acid could ameliorate these difficulties. OBJECTIVE: To describe the effect of the folic acid in the cognitive function in epileptic patients who take phenytoin and carbamazepine. METHODS: We chose patient who have epilepsy and that are being treated with phenytoin, carbamazepine or both and formed two groups. The study group was treated with a daily dose of 5 mg of folic acid and the control group was administered placebo for a period of six months, with nine patients in each group of same age, sex, education level, epilepsy's evolution, frequency of seizures, EEG abnormalities and antiepileptic drugs plasma levels. We registered data at the beginning (basal) and at the end of the study. RESULTS: Measurements of basal folic acid plasma levels in both groups were under the referential value. The neuropsychological assessment at the beginning (Mini Barcelona test) showed a deficit in the verbal memory skills in both groups. After six months of treatment with folic acid (study group), the folic acid plasma level was 12.2 mg/mL (p < 0.01) higher than the basal value. Verbal memory test has improved with respect to the basal value (p < 0.05). The numbers of seizures and the plasma levels of the antiepileptic drugs remained unchanged. On the other hand, the group treated with placebo did not improve. CONCLUSION: Treatment with folic acid is safe and without side effects, it improved the cognitive function in patients with epilepsy treated with phenytoin and carbamazepine. PMID- 16315637 TI - [Prevalence of dental fluorosis and additional sources of exposure to fluoride as risk factors to dental fluorosis in schoolchildren of Campeche, Mexico]. AB - OBJECTIVE: To determine the prevalence and severity of dental fluorosis, and to evaluate supplementary fluoride sources as potential risk factors to fluorosis in school children aged 6-9. MATERIAL AND METHODS: A cross-sectional study was carried out in 320 children attending elementary schools protected by a public preventive dental program in the city of Campeche, Mexico. A self-administered questionnaire directed to the mothers was delivered through the schools and collected in the same way. The examiners were trained and calibrated (kappa > 0.90) in modified Dean's Index. The fluorosis community index (FCI) was calculated. Bivariate analyses were made with Chi2 test; odds ratios (OR) and 95% confidence intervals were calculated. Logistic regression was used in the final model. RESULTS: Fluorosis prevalence was 56.3%, with very mild fluorosis present in 45% of children, mild in 10%, and severe in just 1.3%. The FCI was 0.7. The multivariate model showed that the effect of the supplementary fluoride sources was different between children that started brushing with toothpaste before two years of age (OR = 6.15; IC 95% = 2.03-18.67) and after (OR = 2.14; IC 95% = 1.16 3.94). CONCLUSIONS: Fluorosis prevalence was high for mild modalities, and low for more severe levels. According to FCI the dental fluorosis constitute a public health problem in the studied sample. Exposure to diverse fluoride sources -above and beyond the fluoridated salt program- was a risk factor for dental fluorosis in this community. Results suggest that toothpaste use in children two years of age and younger should be cautious, limited to follow current guidelines, and supervised by parents to minimize the risk of dental fluorosis. PMID- 16315638 TI - [Clinical-pathology correlation aptitude of pneumopathology in undergraduate medical students]. AB - OBJECTIVE: 1) To develop and validate an instrument to evaluate the clinical pathology correlation aptitude in pneumopathology (CPCAP). 2) Compare two groups of students: those who had already taken the respiratory pathology course and students that hadn't. METHODS: An instrument with real anatomopathology respiratory cases was developed. The measurement instrument was validated through expert rounds. It included 116 items of the "true", "false", or "don't know" type, with an agreement of 80 % or more in the answerers between the judges. The internal consistency was determined with Spearman Brown proficiency, reaching a value of 0.78. The sample studied was all the groups of each hospital of an organic pathology undergraduate universitary course. Two sampled were taken, students who had already been through respiratory pathology (Group A), and students who hadn't (Group B). RESULTS: The sample studied was of 485 applicants, from 64 different universitary groups. These were divided in two groups, with 245 (A) and 240 (B) students each. Its global medians were 51 and 36 for groups A and B respectively (p < 0.001). In the 55 % of the subgroups when compared individually taking into account each one of the hospital where they study the test results were significantly better in group A. DISCUSSION: In a general way, students in group A show a major development into CPCAP than students in B. However, development is not enough to reach the expected by the educative program. The results suggest that the teachers put greater effort in the correlation between anatomopathological changes and the clinical and paraclinical manifestations of the patients. PMID- 16315639 TI - [Ligase chain reaction testing of pooled urine specimens to diagnose Chlamydia trachomatis infection]. AB - OBJECTIVE: The purpose of this study was to assess the utility and validity of pooling urine samples for molecular diagnosis of Chlamydia trachomatis infection. MATERIAL AND METHODS: Of 1,220 urine samples collected from Mexican female and male adolescents, 305 pools were composed of fourth individual samples each, based on a calculation of optimal pool size. These were processed by ligase chain reaction (LCR) for the detection of C. trachomatis. Positive and gray-zone pools were reanalyzed individually. Cost savings were calculated comparing actual costs of testing to the cost that would have been incurred testing all 1,220 samples individually. RESULTS: Pools results were: 56 positive, 19 gray-zones and 230 negative. Following individual retesting of positive and gray-zone pools, 59 cases of C. trachomatis infection were identified (4.8% prevalence). Thus, a total of 601 LCR tests were performed, for a 50.4% savings considering only the direct cost of the test. CONCLUSIONS: Our experience shows that sample pooling is both a reliable and convenient tool for CT surveillance in our setting. It should be considered in other similar settings where limited resources constraint surveillance of STIs. PMID- 16315640 TI - CD4 and CD8 T cell response to the rHSP60 from Klebsiella pneumoniae in peripheral blood mononuclear cells from patients with ankylosing spondylitis. AB - OBJECTIVE: To determine the processing pathways used by peripheral blood mononuclear cells (PBMC) and present the rHSP60Kp, and the T cell subpopulations involved in the response, in patients with ankylosing spondylitis (AS) METHODS: The lymphoproliferative response to the rHSP60Kp in PBMC from 14 HLA-B27+ AS patients and 15 B27- healthy controls was assessed by 3H-TdR incorporation. The processing pathways for the rHSP60Kp were analyzed by 3H-TdR incorporation in fresh PBMC from patients using homologous PBMC preincubated with the antigen and specific inhibitors: chloroquine, N-acetyl-L-leucil-L-leucil-L-nor-leucinal (LLnL) or brefeldin A (BFA), fixed with p-formaldehyde (fixed APC). The CD4+/CD8+ T cell subpopulation activated with the antigen was determined by three colours flow cytometry in PBMC from patients. RESULTS: Eight out of fourteen patients showed positive lymphoproliferative responses to the rHSP60Kp while none of the healthy controls responded (p < 0.012). In five patients S.I. was above 4.0. In these patients lymphoproliferation was lower when chloroquine and LLnL was used and it became negative with BFA, indicating that both pathways are used. CD4+ and CD8+ T cells populations expressed CD69 when activated by the rHSP60Kp. CONCLUSIONS: Our results suggest that CD4 and CD8 T cells participate in the response to the rHSP60Kp in B27+ AS patients. PMID- 16315641 TI - Activated protein C resistance and lupus anticoagulant activity induced by plasma and purified monospecific human IgG anti-beta2-glycoprotein-I antibodies. AB - INTRODUCTION: We investigated the activated protein C resistance (APCR) phenotype and the lupus anticoagulant (LA), activity induced by anti-beta2-glycoprotein-I (anti-beta2GP-I) antibodies. PATIENTS AND METHODS: We studied plasma and sera samples from 29 patients with persistently positive anti-beta2GP-I: 22 with thrombosis (12 with primary APS, 10 with APS secondary to SLE) and seven without thrombosis (all with SLE); 25 healthy subjects were studied as controls. We detected anticardiolipin antibodies (ACA); IgG (and its subclasses) and IgM anti beta2GP-I, on irradiated and non-irradiated plates by ELISA. APCR was assessed by the activated partial thromboplastin time (APTT)-based assay and by the modified test. The FV Leiden mutation was studied by PCR. LA determination included screening and confirmatory dRVVT. Serum anti-2 GP-I were affinity purified on sepharose columns and their isotype, subclass, and reactivity against various antigens were studied by ELISA. RESULTS: We found that titers of IgG anti-beta2GP I on irradiated plates were higher than on non-irradiated plates (p = 0.002), IgG2 was the predominant subclass. Fifteen patients (13 with thrombosis) had LA and 15 (also 13 with thrombosis) induced the APCR phenotype. Eleven (all with thrombosis) had both. Two patients were heterozygous for the Leiden mutation. Two purified antibodies, monospecific for beta2GP-I, induced an in vitro APCR phenotype and LA activity. CONCLUSIONS: Our results seem to indicate that the inhibition of the APC anticoagulant function by IgG2 anti-beta2GP-I with LA activity may be one of the responsible mechanisms of thrombophilia in patients with APS. PMID- 16315642 TI - [Genomic retinoblastoma perspectives: implications of tumor supressor gene RB1]. AB - In order to define the molecular and cellular bases of the development of retinoblastomas it is necessary to know its etiology, and to apply the advances in genome technology to this kind of neoplasia. Retinoblastomas are childhood tumors of the eye with an average incidence of one case in every 15,000-20,000 live births, which occur in sporadic and hereditary forms. The sporadic form appears regularly as a unilateral tumor, while in the familial form of the disease, tumors may be unilateral and bilateral. This neoplasia is characterized by leukocoria, strabism, and heterochromia. The retinoblastoma gene (RB1) is a molecular marker of retinoblastoma tumors. This gene is located in chromosome 13q14.2 and encodes a nuclear phosphoprotein (pRB) of 110 KDa, which plays a major role in cell proliferation control through cell cycle-regulated phosphorylation/dephosphorylation cycles of this protein. The RB1 gene is mainly affected by point mutations, which occur most frequently in exons 3, 8, 18 and 20. At the end of the last century, DNA technology has improved notably, allowing for its application to the study of a vast array of diseases. The aim of this work is to show the molecular aspects involved in retinoblastoma which are currently deciphering; this is possible thanks to new technology platforms that have been developed. This will allow us in a near future, to offer tests for the early diagnoses, prognoses, and the determination of individual predisposition towards this neoplasia. PMID- 16315643 TI - [Virus, immunosuppression, and the kidney transplant recipient]. PMID- 16315644 TI - [Pathogenesis of portal hypertension]. AB - It is now well established that portal hypertension is not a purely mechanical phenomenon. Primary hemodynamic alterations develop in the hepatic and systemic circulatory systems; these alterations in combination with mechanical factors contribute to the development of portal hypertension. In the hepatic circulation, these hemodynamic alterations are characterized by vasoconstriction and impaired hepatic vasodilatory responses, whereas in the systemic circulation, particularly in the splanchnic bed, vessels are hyperemic with increased flow. Thus, an increase in intrahepatic resistance in conjunction with increased portal venous inflow, mediated through splanchnic dilation, contributes to the development of portal hypertension. The ensuing development of elevated flow and transmural pressure through collateral vessels from the hypertensive portal vasculature into the lower pressure systemic venous circulation accounts for many of the complications, such as bleeding esophageal varices, observed with portal hypertension. The importance of the primary vascular origin of portal hypertension is emphasized by the utility of current therapies aimed at reversing these hemodynamic alterations, such as nitrates, which reduce portal pressure through direct intrahepatic vasodilatation, and ,B blockers and octreotide, which reduce splanchnic vasodilatation and portal venous inflow. New evidence concerning relevant molecular mechanisms of contractile signaling pathways in hepatic stellate cells and the complex regulatory pathways of vasoactive molecules in liver endothelial cells makes a better understanding of these processes essential for developing further experimental therapies for portal hypertension. This article examines the current concepts relating to cellular mechanism that underlie the hemodynamic alterations that characterize and account for the development of portal hypertension. PMID- 16315645 TI - [Tobacco and tobacco use disorder in the history of Mexico and Europe]. AB - The tobacco used for a long time by American and Caribbean natives was introduced into Europe at the end of XV century, and essentially during the XVI century, by travelers returning to their Fatherlands. After the tobacco industry was organized, several concerns arose regarding medical and social care for the workers in the tobacco factories. Medical and hygienic aspects were reflected in a whole chapter (the XVII) of the Ramazzini's Treatise on Medicine of Work, published in 1700. Concerning social care for the workers' families, the creation, April 1796, of nursery schools for the children of working women in tobacco factories of the New Spain must be recalled. In opposition to the predictions of some natives and visitors during the last centuries, with the passage of time, the tobacco habit instead of decreasing, became progressively more accentuated in all social classes. To aggravate conditions, at present, the noxious effects of the tobacco smoke are combined with those of environmental contamination. Recent epidemiological reports on the number and health conditions of smokers, as well as the National Antitobacco Program in Mexico, are mentioned. PMID- 16315646 TI - Direct likelihood analysis versus simple forms of imputation for missing data in randomized clinical trials. AB - BACKGROUND: In many clinical trials, data are collected longitudinally over time. In such studies, missingness, in particular dropout, is an often encountered phenomenon. METHODS: We discuss commonly used but often problematic methods such as complete case analysis and last observation carried forward and contrast them with broadly valid and easy to implement direct-likelihood methods. We comment on alternatives such as multiple imputation and the expectation-maximization algorithm. RESULTS: We apply these methods in particular to data from a study with continuous outcomes. The outcomes are modelled using a general linear mixed effects model. The bias with CC and LOCF is established in the case study and the advantages of the direct-likelihood approach shown. CONCLUSIONS: We have established formal but easy to understand arguments for a shift towards a direct likelihood paradigm when analysing incomplete data from longitudinal clinical trials, necessitating neither imputation nor deletion. PMID- 16315647 TI - Decision rule based multiplicity adjustment strategy. AB - To minimize potential controversies in determining the need for multiplicity adjustment for multiple hypotheses, we propose a decision rule based multiplicity adjustment strategy in this paper. Resorting to a predefined decision rule of a clinical trial, one may link the different hypotheses by their logical relationships and divide them into different families. A proper multiplicity adjustment procedure can then be developed by maintaining strong control of Type I error rate within each family. The paper applies the proposed multiplicity adjustment strategy to a published raloxifene clinical trial. PMID- 16315648 TI - Minority recruitment to the Selenium and Vitamin E Cancer Prevention Trial (SELECT). AB - BACKGROUND: Previous large chemoprevention studies have not recruited significant numbers of minorities. The Selenium and Vitamin E Cancer Prevention Trial (SELECT) is a large phase III study evaluating the impact of selenium and vitamin E on the clinical incidence of prostate cancer. Over 400 SELECT study sites in the USA, Canada, and Puerto Rico recruited men to this trial. The SELECT recruitment goal was 24% minorities, with 20% black, 3% Hispanic, and 1% Asian participants. The goal for black participants was set at 20% because of their proportion in the United States population and their prevalence of prostate cancer. METHODS: The minority recruitment strategies in SELECT were to: 1) consider minority recruitment during site selection; 2) expand the eligibility criteria by lowering the age criterion for black men and including men with controlled co-morbid illnesses; 3) develop a national infrastructure; 4) give additional funds to sites with the potential to increase black enrollment; and 5) provide resources to maximize free media opportunities to promote SELECT. RESULTS: SELECT recruitment began in August 2001 and was intended to last five years, but concluded two years ahead of schedule in June 2004. Of the 35 534 participants enrolled, 21% were minorities, with 15% black, 5% Hispanic, and 1% Asian. CONCLUSIONS: Careful planning, recruitment of large numbers of clinical centers and adequate resources accomplished by the combined efforts of the National Cancer Institute (NCI), Southwest Oncology Group (SWOG), SELECT Recruitment and Adherence Committee (RAC), SELECT Minority and Medically Underserved Subcommittee (MMUS), and the local SELECT sites resulted in attainment of the estimated sample size ahead of schedule and recruitment of the largest percentage of black participants ever randomized to a cancer prevention trial. PMID- 16315649 TI - Maximum information designs. AB - BACKGROUND: Expressions to determine the sample size N needed to provide power 1 beta to detect a difference between groups, say delta, involve other nuisance parameters, such as the variance of the observations for a test or means, or the control group probability for a test of proportions or the control group hazard rate for a logrank test of event-times. Designs where N is fixed are called maximum N or duration designs because the sample size and required duration of the study can be fixed, or estimated. However, such designs are expected, but not guaranteed, to provide the desired level of power to detect the specified difference delta at the study end because the true or estimated values of the nuisance parameters are unknown. Thus, the actual information to be accrued and the associated level of power are random variables with sample variation. METHODS: Expressions are developed to determine the amount of information needed to provide the desired level of power, regardless of the values of the nuisance parameters. RESULTS: The amount of information (I) in the observed data is readily quantified and can often be expressed as the inverse of the variance of the test statistic. Also, the total amount of information required to provide the desired level of power 1-beta to detect a difference delta with a test at level alpha is readily determined, designated as Ialpha,beta,delta. Under a maximum information design, the study is continued until the required total amount of information is accrued, or I = Ialpha,beta,delta. In this case, the sample size or duration are random variables, but each can be estimated under various projections about the underlying parameters (variance, probability, hazard) in advance. The implementation of a maximum information design for two and multiple group trials is described for a test of means, proportions and event-times using the logrank test. Application to other methods of analysis is described. CONCLUSIONS: A maximum information design provides greater assurance that the desired level of power will be attained. However, the exact study duration is unknown. Issues related to the implementation of such a design are discussed. PMID- 16315650 TI - There's a great deal of motion (and commotion) but are we going anywhere? PMID- 16315651 TI - Hospital RNs' and CNOs' perceptions of the impact of the nursing shortage on the quality of care. AB - The results of analyses of the 2002 and 2004 National Sample Surveys of RNs and the 2004 National Sample of CNOs document the widespread perception that the shortage of nurses is a major problem for the overall quality of patient care in hospitals. In the eyes of the majority of hospital RNs and CNOs surveyed, care processes involving communication, timely response to pages and telephone calls, delays in patient discharges, and the time patients had to wait for tests and procedures were all affected negatively by the nursing shortage. These results are not encouraging. PMID- 16315652 TI - The importance of level 1 trauma services in U.S. hospitals. AB - In this quantitative research study, the organizational characteristics, market factors, and profitability of U.S. acute care hospitals that provide the highest intensity of trauma services are assessed. Results indicate these hospitals are larger, have a higher occupancy rate, higher expenses per discharge, and a lower length of stay. Hospitals with Level 1 trauma centers have a positive return on assets. The study has managerial implications associated with individual hospital performance and policy implications on resource allocation. PMID- 16315653 TI - Characteristics of internationally educated nurses in the United States. AB - Among an estimated 2.7 million U.S. nurse workforce in March 2000, about 4% were internationally educated nurses. This secondary analysis of the 2000 National Sample Survey of Registered Nurses profiles this unique segment of the U.S. nurse workforce. Findings indicate that internationally educated nurses have distinctive demographical, educational, and employment characteristics when compared to U.S.-trained nurses. Implications of the findings are elaborated in lieu of the present U.S. nurse shortage. PMID- 16315654 TI - Crisis management policies and programs to prevent nursing-related medical disputes in Taiwanese hospitals. AB - Hospital nursing directors' levels of general satisfaction toward nursing-related crisis management and their institutions' overall crisis management policies and programs were investigated. The analyses showed that shorter length of institution operation in years, having more comprehensive assessments, being less comprehensive in the public communication aspect, and having higher levels related to the necessity of the public communication aspect contributed to nursing directors having higher general satisfaction levels toward nursing related crisis management. PMID- 16315655 TI - Targeting educational initiatives. AB - The nursing education department of a large teaching hospital faced the challenge of prioritizing the delivery of staff education programs after reductions in personnel and resources. Patient population data were used to guide curricula development. Results included improved clinical practice in a cost-effective manner. PMID- 16315656 TI - Measuring nursing workload in ambulatory care. PMID- 16315657 TI - Misery finds collaborative company: addressing the health workforce shortage through a multidisciplinary approach. PMID- 16315658 TI - In search of a reason to influence health policy. PMID- 16315659 TI - Establishing standards for intranet on-line education. AB - Following a clear set of on-line programming standards assists educators with consistency the training expectations from participants. Placing staff educational materials and health information online is an effective way to disseminate information and an efficient method of providing ongoing staff education. The presentation of on-line programming should be organized, comprehensive, and up-to-date. In addition, on-line programming should also be appealing, dynamic, and, above all practical. Today's clinical staff are pushed to the limits of time and resources. On-line training is one way to help staff meet their learning needs in today's reality. The inception of the UWMF on-line educational programs began in January 2002. Since that date employee participation has increased steadily. The time for Intranet on-line program development is part of the staff education department. For example, one area of programming is the mandatory educational credits needed for the organizations 100 certified medical assistants (CMAs). Figure 1 shows the nearly 3.5 years (January 2002 through May 2005) of participation. The 912 sessions made available 1,870 CEs for the CMAs. If the organization's CMAs were to pay for the continuing education credits (CEs) outside of the clinic setting, it could cost on average of dollar 10.00 per CE, or dollar 18,700 during the same 3-year period. Another aspect of economics is the cost in time away from work. On-line education allows employees to participate when time is available, versus having to leave work when traditional educational inservices are being held. This flexibility in participation can lead to better staff coverage. PMID- 16315660 TI - The Johnny Appleseeds of organizational change. PMID- 16315661 TI - Pathogens are the most frequent of frequent flyers. PMID- 16315662 TI - Exotic animal diseases bulletin. Trichinella. PMID- 16315663 TI - Clinicopathological findings associated with feline infectious peritonitis in Sydney, Australia: 42 cases (1990-2002). AB - OBJECTIVES: To review the clinicopathological findings in naturally-occurring, histopathologically confirmed cases of feline infectious peritonitis in client owned cats in Sydney, Australia, with the purpose of identifying factors assisting in the diagnosis of this complex disease syndrome and to characterise the disease as it occurs in this region. DESIGN: Retrospective clinical study: the clinical records of all cats with histopathologically confirmed feline infectious peritonitis at the University Veterinary Centre Sydney and a private cat hospital in Sydney between 1990 and 2002 were reviewed for signalment, history, physical findings, diagnostic test results and the distribution of histological lesions throughout the body at necropsy. RESULTS: Forty-two cats met the inclusion criteria. Significant features of this study that unique to the contemporary literature are i) the over-representation of certain breeds (Burmese, Australian Mist, British Shorthaired, and Cornish Rex) and the under representation of other breeds (Domestic Shorthaired, Persian); ii) the overrepresentation of males; iii) the tendency for effusive disease in Australian Mist cats and non-effusive disease in Burmese; iv) the even age distribution of disease seen in cats older than 2 years-of-age; and v) the presence of fulminant immune-mediated haemolytic anaemia in two cats in this study. CONCLUSION: The study highlights the diverse range of clinical manifestations and the complexities experienced by clinicians in diagnosing this fatal disease. Some aspects of the epidemiology and clinical manifestations of feline infectious peritonitis appear different to the disease encountered in Europe and North America, most notably the over-representation of specific breeds and the presence of immune-mediated haemolytic anaemia. PMID- 16315664 TI - Traumatic cerebral partial lobotomy in a Thoroughbred stallion. PMID- 16315665 TI - Right forelimb lameness in an 8-year-old rottweiler. PMID- 16315666 TI - Putative sporidesmin toxicity in an Eastern Grey kangaroo (Macropus giganteus). AB - A 2-year-old, captive, male Eastern Grey kangaroo (Macropus giganteus) died after progressive weight loss over a 4 week period. Biochemical analysis suggested hepatobiliary injury. At necropsy the liver was small, pale and firm. There were no abnormalities detected in other organs. Histopathological examination revealed a severe, diffuse, obliterative cholangiohepatopathy with advanced periportal fibrosis. This chronic hepatotoxicity was consistent with exposure to sporidesmin, the toxic metabolite in the spores of the fungus Pithomyces chartarum. Restricted grazing opportunities and heavy fungal pasture contamination may have precipitated sporidesmin toxicity in this animal. Sporidesmin toxicity has not previously been reported in this species. PMID- 16315667 TI - Subtotal agenesis of the cerebellum in a dog. PMID- 16315668 TI - Recent veterinary graduates over the last five decades: recollections and perceptions. AB - OBJECTIVE: To describe the attitudes of veterinary graduates over the last 50 years to their experiences as recent graduates DESIGN: A questionnaire, sent by mail. PROCEDURE: A questionnaire seeking information on experiences as a recent graduate was sent to about 100 veterinarians who graduated in or about 1950, 1960, 1970, 1980, 1990 and 2000, and 68% responded. Data were entered onto an Excel spreadsheet, and analysed with the SAS System 8 for Windows. RESULTS: Most veterinarians who graduated in 1970 and earlier were generally satisfied with their conditions, even though these were often taxing. In the 1970s the cattle industry faltered and rural practice suffered a downturn. Veterinary positions became scarce, and there were few opportunities for graduates to move on from an unsatisfactory workplace--as many were at that time. By 1980 many recent graduates were questioning their decision to study veterinary science. Many of them, working long hours under often stressful conditions, felt that that they were not receiving adequate help or support--or remuneration--from their employers, or respect from their clients. This also afflicted many who graduated in 1990, but by 2000 graduates were reporting an improvement. Their hours of work and the expectations placed on them were more manageable, and the level of help and support, and remuneration, were more acceptable. As a result they found their first year much more enjoyable than had those who had graduated since about 1970. Many of the factors contributing to this improvement resulted from changes in society generally. Specifically, both graduates and employers have become more conscious of what might reasonably be expected, an Award specified minimum levels, the AVA started accrediting New Graduate Friendly Practices, publishing a most informative New Graduate Guide, and helping in other ways. Remuneration, especially when compared with comparable professions, continues to be a source of discontent. Furthermore, despite improved conditions on average, many graduates still suffer under substandard conditions, and these can have an adverse effect on their health and their attitude to their career in veterinary science. CONCLUSIONS: Average conditions of employment of recent veterinary graduates have improved somewhat after a nadir in the 1970s and 1980s, but they are still substandard for some. Remuneration remains the greatest source of dissatisfaction for recent graduates. PMID- 16315669 TI - Mass screening for antimicrobial resistant Escherichia coli in dairy cows in northern New South Wales. AB - OBJECTIVE: To describe aspects of the epidemiology of antimicrobial resistance in Escherichia coli shed in the faeces of milking cows in a dairying region of New South Wales. DESIGN: A survey based on multi-stage sampling with repeated measures made within herds for estimating within-herd correlation of resistance status, and with repeated measures made on identical specimens for estimating test-retest reliability. PROCEDURE: From a population of 110 dairy herds, 30 were selected at random and from each herd between 5 and 10 faecal specimens were obtained from fresh manure pats. E coli from faecal specimens were grown on hydrophobic grid membrane filters (HGMF) and replicated onto chromogenic agar and agar containing antimicrobials (gentamicin, ampicillin, tetracycline and sulfamethoxazole). Image analysis was used to assess colony growth. Data were analysed descriptively, by generalised linear mixed models and by Taylor series linearisation to account for attributes of the survey design. RESULTS: Of the 10,279 E coli isolates assessed, 91% expressed no resistance, 7.3% were resistant to sulfamethoxazole, 3.6% to tetracycline, 2.2% to ampicillin and 0.09% to gentamicin. The most common multiple resistance phenotype was ampicillin tetracycline-sulfamethoxazole (1.8% of isolates). Most multiple resistant isolates appeared clustered within particular herds but were too rare to obtain valid estimates of variance, confidence intervals or intra-herd correlation. The estimated proportion of isolates in the population that were susceptible to all four antimicrobials was 97% (95% CI: 91% to 100%) and 55% of cows had no resistance detected in faecal E coli (95% CI: 27% to 83%). Within-herd correlation of shedding status (any resistance pattern) was absent and test retest reliability of the measurement system was estimated to be at the lower end of good (0.40) but increased to excellent (0.89) after excluding sulfamethoxazole resistance, which had a greater measurement error. CONCLUSION: Antimicrobial resistance was uncommon in E coli in the population of dairy cows studied. HGMF and image analysis is an effective tool for detecting rare forms of resistant E coli that are not uniformly distributed in livestock populations. PMID- 16315670 TI - Some physiological responses associated with reduced wool growth during blowfly strike in Merino sheep. AB - OBJECTIVE: To examine the effect on wool growth of physiological changes associated with experimental flystrike in superfine Merino wethers. DESIGN: An animal house study comparing experimentally fly struck sheep with non-infected control groups that were pair fed or fed ad libitum. PROCEDURE: Sheep in the blow fly larval challenge group were each artificially infected with 500 first instar larvae per day for 8 days. Infections were terminated with insecticide on day 10. For all sheep, live weight change, feed intake and body temperature were monitored and blood collected for analysis of haematology, cortisol, serum haptoglobin, serum amyloid A and the pro-inflammatory cytokines, TNFa, IL-1 b, IL 6 and IL-8. Wool growth and staple strength were examined 4 months after infection. RESULTS: Experimental fly strike included moderate fever, depressed feed intake, and elevated cortisol, IL-6, serum amyloid A and haptoglobin. Staple strength was significantly decreased in struck sheep and did not differ between wool from sites adjacent to and remote from strike lesions. CONCLUSIONS: The results suggest that depression in feed intake alone is not sufficient to account for the decline in staple strength seen during fly strike. Cortisol, IL-6 and metabolic consequences of the acute phase response are likely to be major contributors to the systemic effects of blowfly strike on wool. PMID- 16315671 TI - Idiopathic thrombocytopenia in Cavalier King Charles Spaniels. AB - OBJECTIVE: To determine the prevalence of asymptomatic idiopathic macrothrombocytopenia in the population of Cavalier King Charles Spaniels (CKCS) in New South Wales (NSW) and to determine if it exhibits an autosomal recessive inheritance pattern. We also aimed to determine if significant differences existed when counting platelets manually, by auto analyser or by blood smear estimation in CKCS and mixed breed dogs. METHODS: Blood was collected from 172 dogs (152 CKCS and 20 mixed breed) and placed into sodium-citrate anticoagulant. Platelet counts were performed manually, by auto analyser and by blood smear estimates in CKCS and mixed breed dogs. Blood smears were also examined for platelet clumping and erythrocyte, leukocyte and platelet morphology. Pedigree analysis was performed to determine if an autosomal recessive inheritance pattern was supported. RESULTS: A statistically significant difference was found in platelet counts between CKCS and mixed breed dogs (P < 0.0001). CKCS had a platelet count that was 32% that of the controls (95% confidence interval, 28 to 37%). There was no significant difference between methods used to count platelets. Thirty percent of CKCS had macrothrombocytes. Pedigree analysis and examination of obtained and expected segregation ratios from 17 CKCS families supported an autosomal recessive pattern of Mendelian inheritance. CONCLUSIONS: A high prevalence of idiopathic macrothrombocytopenia exists in CKCS in NSW and automated or blood smear estimates are sufficient to count platelet numbers. Data supports an autosomal recessive inheritance pattern. PMID- 16315674 TI - Seeing the big picture. PMID- 16315675 TI - What's next? PMID- 16315676 TI - Why "you snooze, you lose" doesn't work for nurses. PMID- 16315678 TI - 101 ways to improve nursing culture: development of others. PMID- 16315679 TI - Weaning: confusing messages from the Department of Health. PMID- 16315680 TI - Children with a prisoner in the family. PMID- 16315681 TI - Maternal nutrition in pregnancy and the first postnatal year--2. After the birth. AB - In the second of a two-part article, a senior research scientist discusses the mother's nutritional needs during lactation and the first year after the birth. The mother's needs for energy increase during breast-feeding, but conversely studies have shown that breast-feeding mothers do not necessarily lose weight. As in pregnancy, it is recommended that all lactating women should take a supplement that provides 10 microg per day of vitamin D. Healthy diets for breastfeeding women are essentially the same as for other adults. Parents are often receptive to advice at this time and this is a good opportunity for health professionals to encourage breast-feeding and a healthy diet as this may promote future health. PMID- 16315682 TI - Twins and more--1. Some current thinking on multiple births. AB - The incidence of multiple births is rising in many countries including the U.K., mainly as a result of treatment for infertility. Parents of twins, triplets or more require specific information, advice and support at all stages of parenting from diagnosis of a multiple pregnancy onwards. Specialist midwives have an important role to play in this, as do voluntary organisations such as the Multiple Births Foundation. The first part of this two-part article provides an update of some current thinking on multiple births, including twin-twin transfusion syndrome. Part two will provide information about the needs of families with a multiple birth in the early years. PMID- 16315683 TI - How to take a sexual history. AB - Under the National Strategy for Sexual Health and HIV, most patients seeking or requiring routine sexual health care are now offered the option of being treated by the primary health care team, rather than a specialised genito-urinary medicine clinic. Taking a sexual history and making a risk assessment is a key skill for making a diagnosis and care plan. This article offers a structured approach to this task, particularly for nurses, midwives and other community health professionals. It also describes the often sensitive core questions that the professional may need to ask in order to obtain an effective sexual history and determine the risks for a particular patient. PMID- 16315685 TI - Drug cupboard keys. PMID- 16315684 TI - Why did we bother? PMID- 16315686 TI - Incidentally. PMID- 16315687 TI - I find myself faced with the unhappy decision to resign after 42 years of membership. PMID- 16315688 TI - Non-heart beating organ donation. A case study. AB - The aim of this case study is to discuss the clinical management of a non-heart beating organ donor. This case study will concentrate in particular on the clinical assessment of the potential donor patient undertaken by the donor transplant coordinator (DTC) and the donation process up to the time of transplantation. The author will also describe the differences between heart beating and non-heart beating donors and discuss how transplantation can benefit renal recipient patients. PMID- 16315689 TI - Mentoring opens doors. AB - This article is based on the Daisy Ayris lecture given at the 2003 Congress by Sheila Allen, former Association of Perioperative Registered Nurses president. In it she explores the history and definition of mentoring. Mentors may be persons of rank, achievement and prestige, or they may be a knowledgeable colleague or teacher. They serve as a catalyst to transform as they instruct, counsel, guide and facilitate the development of others. PMID- 16315690 TI - Has not passed urine. But is the bladder full? AB - Is bladder volume of concern to the perioperative team when the patient is undergoing a procedure not related to the same anatomical area? A common scenario is that the patient is requested to void prior to leaving the ward and their word taken that they have done so. The patient then enters the theatre environment and the only time that urinary function is taken into account is if the blood pressure falls and kidney function needs to be monitored. PMID- 16315691 TI - A swab story. AB - This is a tale of when radio-opaque strips in surgical swabs aren't always radio opaque. Sue Lamont has a 30-year background as a theatre nurse and a theatre manager. Her current role is clinical procurement and risk management coordinator for Surgical Services at the University Hospitals of Leicester NHS Trust. A recent experience illustrated her job's many challenges. PMID- 16315692 TI - Space-time analogy for partially coherent plane-wave-type pulses. AB - In this Letter we extend the well-known space-time duality to partially coherent wave fields and, as a limit case, to incoherent sources. We show that there is a general analogy between the paraxial diffraction of quasi-monochromatic beams of limited spatial coherence and the temporal distortion of partially coherent plane wave pulses in parabolic dispersive media. Next, coherence-dependent effects in the propagation of Gaussian Schell-model pulses are retrieved from that of their spatial counterpart, the Gaussian Schell-model beam. Finally, the last result allows us to present a source linewidth analysis in an optical fiber communication system operating around the 1.55 microm wavelength window. PMID- 16315693 TI - Large optical photon sieve. AB - A photon sieve with 10(7) holes has been constructed for operation at optical wavelengths. Details of the design, fabrication, and performance of this device are presented. The 1 m focal-length, 0.1 m diameter element is diffraction limited over a significant bandwidth and has a moderate field of view. PMID- 16315694 TI - Ultrafast pulse retiming by cross-phase modulation in an anomalous-dispersion polarization-maintaining fiber. AB - Retiming of signal pulses by orthogonally polarized control pulses copropagating in an anomalous-dispersion low-birefringence polarization-maintaining fiber is demonstrated. Through walk-off-free cross-phase modulation in a 500 m long fiber, optical pulses at a 10 GHz rate are retimed by as much as +/- 2 ps. PMID- 16315695 TI - Wavelength-versatile subpicosecond pulsed lasers using Raman gain in figure-of eight fiber geometries. AB - A family of compact pulsed fiber lasers is described that employs broadband, wavelength-flexible Raman scattering in passively mode-locked figure-of-eight fiber laser geometries. Specifically, sources at 1.57, 1.33, and 1.41 microm with respective soliton durations of 440, 500, and 860 fs are reported. Operation is possible at other wavelengths with a suitable pump source, gain fiber, and components. PMID- 16315696 TI - Compensation for multimode fiber dispersion by adaptive optics. AB - Adaptive optics is used to compensate for modal dispersion in digital transmission through multimode fiber (MMF). At the transmitter, a spatial light modulator (SLM) controls the launched field pattern. An estimate of intersymbol interference (ISI) caused by modal dispersion is formed at the receiver and fed back to the transmitter, where the SLM is adjusted to minimize ISI. Error-free transmission of 10 Gbit/s non-return-to-zero signals through standard 50 microm graded-index MMFs up to 11.1 km long is demonstrated. It is shown that a single SLM can compensate for modal dispersion across a 600 GHz bandwidth. PMID- 16315697 TI - Investigation of dual-channel fiber-optic surface plasmon resonance sensing for biological applications. AB - A dual-channel fiber-optic sensor based on surface plasmon resonance (SPR) for self-referencing refractive-index measurements has been proposed. Most applications of fiber-optic SPR sensors are designed to measure the refractive index of a liquid or gas sample by measuring the signal from a single surface, the sensitivity and stability of which is easily affected by the fluctuation of external environmental conditions. We have designed a dual-channel fiber-optic surface sensor with two independent SPR signals from two areas of the same probe. A prototype sensor was fabricated and characterized. The preliminary experimental results demonstrate the characteristic responses of both SPR signals from two channels that independently correspond to the refractive index changes in the liquid samples with which they are in contact. The design could be extended to a multichannel sensor with further developments. The experimental results confirmed that one channel can be used as a reference sensor that could compensate for unexpected changes in bulk refraction or temperature and develop this sensor as a practicable high-sensitivity biosensing device. PMID- 16315698 TI - Efficient antiguiding of TE and TM polarizations in low-index core waveguides without the need for an omnidirectional reflector. AB - While an omnidirectional bandgap is necessary to reflect arbitrary polarized light with a planar periodic reflector at any angle of radiation incidence, we demonstrate that omnidirectionality is not at all necessary for efficient antiguiding of modes of any polarization in low-index core photonic bandgap (PBG) fibers and waveguides. For a given radiation decay rate into the reflector and multilayer bandgap size we characterize the phase space of fiber materials leading to the same effective guiding conditions. We demonstrate that low- and high-index-contrast PBG waveguides can antiguide problematic TM-like modes equally effectively. Finally, the possibility of TE-like guidance of a TM polarized mode is described. PMID- 16315699 TI - Thermally switchable and discretely tunable comb filter with a linearly chirped fiber Bragg grating. AB - We propose and experimentally demonstrate a novel switchable and discretely tunable comb filter based on a thermally induced linearly chirped fiber Bragg grating. Experimentally we achieved a thermally induced optical bandpass filter that has eight switchable passband peaks with peak separations of 1.6 and 3.2 nm, a very narrow bandwidth (as small as 10 pm) of each peak, a tunable range of 16.5 nm, and a greater than 25 dB rejection ratio. Two spectral peaks separated by only 0.8 nm were also obtained with this comb filter. This filter provides the unique advantages of an all-fiber structure, switchable spectral peaks, independent tuning of the center wavelength and wavelength spacing of the spectral peaks, low polarization sensitivity, simple structure, ease of fabrication, and low cost. PMID- 16315700 TI - High-power tunable single-frequency single-mode erbium:ytterbium codoped large core fiber master-oscillator power amplifier source. AB - We present a cladding-pumped single-frequency, single-mode erbium:ytterbium codoped fiber master-oscillator power amplifier source generating up to 151 W of continuous-wave output power at 1563 nm with 33% slope efficiency and 20 dB gain. This source was also tunable and had a stable operation range of 1546 to 1566 nm at an output power level in excess of 125 W. The doped fiber exploited a large core design for improved power handling and mitigation of stimulated Brillouin scattering. There was no sign of having stimulated Brillouin scattering even at the highest power. Despite a large core (V = 12), the output beam was nearly diffraction limited (M2 = 1.1). The source showed slight rollover at over 100 W of output power because of the onset of emission from ytterbium, centered at 1060 nm. PMID- 16315701 TI - Simultaneous measurement of refractive index and temperature based on a reflection-mode long-period grating and an intrinsic Fabry-Perot interferometer sensor. AB - We have demonstrated a single fiber probe for simultaneous measurement of external refractive index and temperature based on two interferometers: a reflection-mode long-period grating (LPG) for refractometry and an intrinsic Fabry-Perot interferometer (IFPI) for temperature measurement. Since the output signal of the combined structure is a superposition of signals produced by both sensors, which have different spatial frequencies, the original signals needed for the index and temperature measurements can be separated and recovered through digital filters. PMID- 16315702 TI - Spectral hole burning for wideband, high-resolution radio-frequency spectrum analysis. AB - We present experimental results for what is to our knowledge the first spectral hole-burning based rf spectrum analyzer to cover 10 GHz of rf analysis bandwidth. The rf signal of interest is modulated onto an optical carrier, and the resultant optical sidebands are burned into the inhomogeneously broadened absorption band of a Tm3+:YAG crystal. At the same time a second, frequency-swept laser reads out the absorption profile, which is a double-sideband replica of the rf spectrum, and thus the rf spectrum can be deduced after spectral calibration of the nonlinear readout chirp. This initial demonstration shows spectral analysis covering 10 GHz of bandwidth with >5500 spectral channels and provides 43 dB of dynamic range. PMID- 16315703 TI - No-loss bent light pipe with an equiangular spiral. AB - Bending a light pipe and maintaining the required volume and size, while at the same time preserving source energy and minimizing bend losses, is the basic criterion for designing a light pipe. An important requirement is the design of a light pipe that is capable of being bent without loss of light and with no limitations on acceptance angle. It is shown that in theory such a light pipe does exist, in a form corresponding to the so-called golden rectangle. Numerical simulations with six different no-loss light pipes with multiple bends are provided as a demonstration. PMID- 16315704 TI - Generalized ray-transfer matrix for an optical element having an arbitrary wavefront aberration. AB - A generalized ray-transfer matrix for describing the action of an optical element having an arbitrary wavefront aberration is obtained. In this generalized ray transfer matrix, the action of the aberrated optical element is represented by the product of radial ray-transfer matrices and tangential ray-transfer matrices. The refraction angle of an incident ray is calculated from the gradient of the wavefront aberration at the point of incidence, and the radial and tangential ray transfer matrices directly use the gradient as a matrix component. To show the validity of the generalized ray-transfer matrix, intercept heights from a spot diagram are calculated with the generalized ray-transfer matrix and compared with those calculated with commercial ray-tracing software. PMID- 16315705 TI - Demonstration of a multiwave coherent holographic beam combiner in a polymeric substrate. AB - We demonstrate an efficient coherent holographic beam combiner (CHBC) that uses angle multiplexing of gratings in a thick polymeric substrate. Our experimental results compare well with the theoretical model based on the coupled-wave theory of multiwave mixing in a passive medium. A CHBC of this type may prove useful in producing a high-power laser by combining amplified beams produced by splitting a master oscillator. Furthermore, the ability to angle multiplex a large number of beams enables a CHBC to be used in multiple-beam interferometry applications as a high-precision surface sensor. PMID- 16315706 TI - Volume holographic optical elements for point-to-point imaging with local cross talk. AB - A novel volume holographic optical element serving as an atypical mirror with a lateral magnification of 1 in both the horizontal and the vertical directions is proposed. Optical imaging is performed by point-to-point imaging with local cross talk, which prevents ghost diffraction spots from the holographic element of multiple gratings. PMID- 16315707 TI - Adaptive monogenic filtering and normalization of ESPI fringe patterns. AB - A technique is presented for filtering and normalizing noisy fringe patterns, which may include closed fringes, so that single-frame demodulation schemes may be successfully applied. It is based on the construction of an adaptive filter as a linear combination of the responses of a set of isotropic bandpass filters. The space-varying coefficients are proportional to the envelope of the response of each filter, which in turn is computed by using the corresponding monogenic image [Felsberg and Sommer, IEEE Trans. Signal Process. 49, 3136 (2001)]. Some examples of demodulation of real Electronic Speckle Pattern Interferometry (ESPI) images patterns are presented. PMID- 16315708 TI - Relative focus map estimation using blind deconvolution. AB - An automatic focus map extraction method is presented that uses a modification of blind deconvolution for estimation of localized blurring functions. We use these local blurring functions [so-called point-spread functions (PSFs)] for extraction of focus areas on ordinary images. In this inverse task our goal is not image reconstruction but the estimation of localized PSFs and the relative focus map. Thus the method is less sensitive than general deconvolution is to noise and ill posed deconvolution problems. The focus areas can be estimated without any knowledge of the shooting conditions or of the optical system used. PMID- 16315709 TI - Single-photon imaging at 20,000 frames/s. AB - A complete two-dimensional imaging system based on a silicon monolithic array of 60 single-photon counters is presented. The fabricated solid-state array is rugged and operates at low voltages. Detection efficiency is higher than 40% in the visible range, and cross talk among 50 microm pixels is lower than 10(-4). The complete system provides a maximum throughput of 20 kframes/s with truly parallel readout and nanosecond gating, thanks to the use of an integrated active quenching circuit for each pixel of the array. We report optical and electrical characterizations of the whole imaging system. PMID- 16315710 TI - Low-power optical vibration detection by photoconductance monitoring with a laser speckle pattern. AB - We present a quantitative evaluation of the performance of a photoconductance monitoring sensor array in a speckle-based vibration detection configuration. The device is found to be capable of detecting nanometer-amplitude vibrations in a single shot with incident intensities of only a few microwatts per square centimeter at kilohertz frequencies. This performance indicates that the photoconductance-monitoring array requires approximately 3 orders of magnitude lower intensity to achieve the same displacement sensitivity as competing technologies, such as photo-electromotive-force detectors. PMID- 16315711 TI - Evidence of thermal effects in a high-power Er3+-Yb3+ fiber laser. AB - We analyze the influence of heat generation caused by nonradiative transitions in a high-power 1.55 microm double-clad erbium-ytterbium fiber laser on the Stark level population. At strong pumping rates, 1 microm lasing can start as a result of parasitic reflections. We present a model that allows us to simulate the effect of self-generated heat on the Stark level population by using the MacCumber relation. Heat generation plays a significant role and improves the 1.5 microm laser's efficiency by increasing the 1 microm lasing threshold. PMID- 16315712 TI - Experimental investigation of an inhomogeneous loss and its influence on multiwavelength fiber lasers. AB - Inhomogeneous loss generated by multimode laser linewidth broadening in an optical fiber is experimentally studied. With this mechanism, multiwavelength lasing is achieved by use of either fiber Raman gain or erbium-doped fiber gain. Through various pump powers and optical filter bandwidths, the relationship between inhomogeneous loss and the performance of a multiwavelength fiber laser is studied, and a physical explanation is provided. PMID- 16315713 TI - Modulation and the linewidth enhancement factor of a diode-pumped Nd:YVO4 laser. AB - A technique based on an FM/AM method is proposed for measuring the linewidth enhancement factor a of a diode-pumped Nd:YVO4 laser. A standard rate equation model is complemented with temperature effects to explain the observed behavior under pump modulation. The result alpha = 0.25 +/- 0.13 agrees with values deduced from recent optical injection experiments. PMID- 16315714 TI - Interactions between spherical nanoparticles optically trapped in Laguerre Gaussian modes. AB - When a Laguerre-Gaussian (LG) laser mode is used to trap nanoparticles, the spatial disposition of the particles about the beam axis is determined by a secondary mechanism that engages the input radiation with the interparticle potential. This analysis, based on the identification of a range-dependent laser induced energy shift, elicits and details features that arise for spherical nanoparticles irradiated by a LG mode. Calculations of the absolute minima are performed for LG beams of variable topological charge, and the results are displayed graphically. It is shown that more complex ordered structures emerge on extension to three- and four-particle systems and that similar principles will apply to other kinds of radially structured optical mode. PMID- 16315715 TI - Evanescent-wave bonding between optical waveguides. AB - Forces arising from overlap between the guided waves of parallel, microphotonic waveguides are calculated. Both attractive and repulsive forces, determined by the choice of relative input phase, are found. Using realistic parameters for a silicon-on-insulator material system, we estimate that the forces are large enough to cause observable displacements. Our results illustrate the potential for a broader class of optically tunable microphotonic devices and microstructured artificial materials. PMID- 16315716 TI - Two-photon autofluorescence spectroscopy and second-harmonic generation of epithelial tissue. AB - A spectroscopy system is developed for studying the two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) of epithelial tissue in backscattering geometry. Our findings show that TPEF signals from epithelial and underlying stromal layers exhibit different spectral characteristics, providing information on the biomorphology and biochemistry of tissue. The SHG signal serves as a sensitive indicator of collagen to separate the epithelial layer from underlying stroma. The polarization dependence of the SHG signal reveals a well ordered orientation of collagen fibers in the stromal layer. The results demonstrate the potential of depth-resolved TPEF and SHG in determining the pathology of epithelial tissue. PMID- 16315717 TI - Gold nanocages as contrast agents for spectroscopic optical coherence tomography. AB - We describe gold nanocages as a new class of potential contrast agent for spectroscopic optical coherence tomography (OCT). Monodispersed gold nanocages of an approximately 35 nm edge length exhibit strong optical resonance, with the peak wavelength tunable in the near-infrared range. We characterized the optical properties of the nanocage by using OCT experiments along with numerical calculations, revealing an absorption cross section approximately 5 orders of magnitude larger than conventional dyes. Experiments with tissue phantoms demonstrated that the nanocages provide enhanced contrast for spectroscopic as well as conventional intensity-based OCT imaging. PMID- 16315718 TI - Fractal mechanisms of light scattering in biological tissue and cells. AB - We use fractal continuous random media to model visible and near-infrared light scattering by biological tissue and cell suspensions. The power law of the reduced scattering coefficient, the anisotropy factor of scattering, and the phase function are derived with good agreement with experimental results. Implications for spectroscopic tissue diagnosis are discussed. PMID- 16315719 TI - Imaging of small nanoparticle-containing objects by finite-element-based photoacoustic tomography. AB - We present semiquantitative photoacoustic images of small nanoparticle-containing objects having a wide range of contrast levels relative to the background. The images are obtained by a finite-element reconstruction algorithm that is based on the Helmholtz-like photoacoustic wave equation in the frequency domain. Our reconstruction approach is an iterative Newton method coupled with combined Marquardt and Tikhonov regularizations that can extract the spatial distribution of relative optical absorption property in heterogeneous media. We demonstrate experimental images in single- and multiple-object configurations with a circular scanning photoacoustic tomographic system. The results obtained show that millimeter-size nanoparticle-containing objects can be clearly detected in terms of position, size, and relative optical properties. PMID- 16315720 TI - Highly efficient third-order optical nonlinearities in donor-substituted cyanoethynylethene molecules. AB - We investigated the third-order nonlinear optical properties of several donor substituted cyanoethynylethene molecules in the zero-frequency limit. We observed nonlinearities that are extraordinarily large relative to the small molecular mass of these molecules and that are within a factor of 50 from the fundamental limit. At a wavelength of 1.5 microm, the rotational average of the third-order molecular polarizability is 53 +/- 13 x 10(-48) m5 V(-2) (3.8 x 10(-33) esu) for the best molecule, which corresponds to 1.7 x 10(-48) m5 V(-2) per delocalized electron. The high nonlinear efficiency of these molecules is due to the compact two-dimensional conjugated system and the effective donor-acceptor substitution patterns. PMID- 16315721 TI - Polarization multiplexed write-once-read-many optical data storage in bacteriorhodopsin films. AB - In polymeric films of bacteriorhodopsin (BR) a photoconversion product, which was named the F620 state, was observed on excitation of the film with 532 nm nanosecond laser pulses. This photoproduct shows a strong nonlinear absorption. Such BR films can be used for write-once-read-many (WORM) optical data storage. We demonstrate that a photoproduct similar or even identical to that obtained with nanosecond pulses is generated on excitation with 532 nm femtosecond pulses. This photoproduct also shows strong anisotropic absorption, which facilitates polarization storage of data. The product is thermally stable and is irretrievable to the initial B state either by photochemical reaction or through a thermal pathway. The experimental results indicate that the product is formed by a two-photon absorption process. Optical WORM storage is demonstrated by use of two polarization states, but more polarization states may be used. The combination of polarization data multiplexing and extremely short recording time in the femtosecond range enables very high data volumes to be stored within a very short time. PMID- 16315722 TI - Generation of a radially polarized laser beam by use of a conical Brewster prism. AB - To generate a radially polarized laser beam we designed and fabricated a new Brewster optical element that consists of convex and concave conical prisms. The lateral surface of the convex conical prism was coated with a dielectric multilayer (SiO2 and Ta2O5) to enhance polarization selectivity. By combining two prisms we obtained a conical Brewster prism without beam divergence owing to refraction. A radially polarized TEM01* (R-TEM01*) mode laser beam was demonstrated when this prism was used inside a Nd:YAG laser cavity. PMID- 16315723 TI - Vertically coupled whispering-gallery-mode resonator waveguide. AB - We theoretically describe a novel configuration of a vertically coupled whispering-gallery-mode resonator waveguide. The waveguide is formed as a chain of doughnut-shaped low-contrast ripples on the surface of a vertical cylinder made from an optically transparent material. One can manipulate the properties of the waveguide by changing the shape of the resonators and the separation distance between them. Application of the structure as an optical delay line is discussed. PMID- 16315724 TI - Tunable asymmetrical Fano resonance and bistability in a microcavity-resonator coupled Mach-Zehnder interferometer. AB - We propose a simple microresonator scheme for a Mach-Zehnder interferometer in which a microresonator is side coupled to one arm and a phase shifter is introduced into the other arm, to produce an asymmetric Fano-resonance line shape. In this system, a phase shifter is used to control the variation of the asymmetric line shape, with another reverse resonance next to a resonance minimum over a very narrow frequency range, which results from the interference between a direct channel and a high-Q resonance indirect channel. We also theoretically investigate the novel bistability characteristic based on these shapes. PMID- 16315725 TI - Coherence characteristics of electrically excited tandem organic light-emitting diodes. AB - A tandem organic light-emitting diode structure, excited electrically in the pulsed domain and confined within a double spatial filter configuration, is observed to emit a low-divergence beam (deltatheta approximately 2.53 mrad, or approximately 1.1 times the diffraction limit) with a near-Gaussian spatial distribution. The emission originates from the laser dye Coumarin 545 T, which is used as a dopant. Spectral coherence was determined by use of a double-slit interferometer. The interferometric distribution from our device approximates the interferometric pattern obtained from well-known lasers emitting at lambda approximately 540 nm. PMID- 16315726 TI - Optical Dyakonov surface waves at magnetic interfaces. AB - We address the existence and properties of lossless surface waves that form at interfaces between magnetic and birefringent media. We show that the angular domain of existence of Dyakonov surface waves for magnetic interfaces is significantly larger than that for nonmagnetic ones. Our results have important implications for the experimental generation of surface waves and for their potential applications based on guided-to-leaky transitions. PMID- 16315727 TI - Beaming of light and enhanced transmission via surface modes of photonic crystals. AB - We report beaming and enhanced transmission of electromagnetic waves by use of surface corrugated photonic crystals. The modes of a finite-size photonic crystal composed of dielectric rods in free space have been analyzed by the plane-wave expansion method. We show the existence of surface propagating modes when the surface of the finite-size photonic crystal is corrugated. We theoretically and experimentally demonstrate that the transmission through photonic crystal waveguides can be substantially increased by the existence of surface propagating modes at the input surface. In addition, the power emitted from the photonic crystal waveguide is confined to a narrow angular region when an appropriate surface corrugation is added to the output surface of the photonic crystal. PMID- 16315728 TI - Influence of propagation and external phase in sequential two-photon absorption of femtosecond pulses. AB - We study the influence of shaping the phase of the optical field when time delayed femtosecond pulse pairs excite a sequential two-photon transition in rubidium atoms. Propagation through this optically dense medium modifies the pulse profile. When an external phase is introduced into one of the pulses, partial cancellation of the two-photon absorption signal is observed for temporal delays much greater than the pulse duration. This behavior is qualitatively explained in terms of a negative group delay. PMID- 16315729 TI - Photon-number fluctuation and correlation of bound soliton pairs in mode-locked fiber lasers. AB - Quantum photon-number fluctuation and correlation of bound soliton pairs in mode locked fiber lasers are studied on the basis of the complex Ginzburg-Landau equation model. We find that, depending on their phase difference, the total photon-number noise of the bound soliton pair can be larger or smaller than that of a single soliton, and the two solitons in the soliton pair have a corresponding positive or negative photon-number correlation. It is predicted for the first time to our knowledge that out-of-phase soliton pairs can exhibit less noise as a result of negative correlation. PMID- 16315730 TI - 10 kHz detection of CO2 at 4.5 microm by using tunable diode-laser-based difference-frequency generation. AB - A compact, high-speed tunable, diode-laser-based mid-infrared (MIR) laser source has been developed for absorption spectroscopy of CO2 at rates up to 10 kHz. Radiation at 4.5 microm with a mode-hop-free tuning range of 80 GHz is generated by difference-frequency mixing the 860 nm output of a distributed-feedback diode laser with the 1064 nm output of a diode-pumped Nd:YAG laser in a periodically poled lithium niobate crystal. MIR absorption spectroscopy of CO2 with a detection limit of 44 ppm m at 10 kHz is demonstrated in a C2H4-air laminar diffusion flame and in the exhaust of a liquid-fueled model gas-turbine combustor. PMID- 16315731 TI - Nonlinear absorption of ultrashort laser pulses in thin metal films. AB - Self-consistent simulations of the ultrafast electron dynamics in thin metal films were performed. A regime of nonlinear oscillations was observed that corresponds to ballistic electrons bouncing back and forth against the films' surfaces. When an oscillatory laser field is applied to the film, the field energy is partially absorbed by the electron gas. Maximum absorption occurs when the period of the external field matches the period of the nonlinear oscillations, which, for sodium films, lies in the infrared range. Possible experimental implementations are discussed. PMID- 16315732 TI - State licensure update: giving voice to the value and vision. PMID- 16315733 TI - Future career paths--is it time to phase out AS-CLT programs? PMID- 16315734 TI - Clinical doctorate in laboratory science. PMID- 16315735 TI - The clinical laboratory practitioner. AB - OBJECTIVE: This study was designed to investigate potential areas of practice for the clinical laboratory scientist (CLS) and to propose a graduate curriculum to prepare the practitioner for an advanced level of practice. DESIGN: Meta-analysis of PharmD, physician assistant, physical therapy, and nurse practitioner curricula focusing on academic and clinical advanced practice was used to develop an educational model and curriculum for a professional doctorate in clinical laboratory science (CLS). MAIN OUTCOME MEASURE: (1) New educational model for CLS advanced practice; (2) A proposed curriculum for a Doctorate of Clinical Laboratory Science degree. RESULTS: A new curriculum model was adapted from established healthcare educational models. CONCLUSION: Although there is a need for a baccalaureate degree in CLS there is also a role for expanded education and responsibilities for CLS practitioners. The CLS Advanced Practitioner design focuses on moving students from the baccalaureate level to the doctoral level and prepares the individual to become an integral part of the healthcare team. PMID- 16315736 TI - Effect of adverse storage conditions on performance of glucometer test strips. AB - OBJECTIVE: A study was conducted to assess the impact of adverse storage environments, i.e., not manufacturer recommended, on the performance of reagent test strips used with a point of care testing (POCT) glucometer to measure whole blood glucose levels. DESIGN/SETTING: Glucose reagent test strips were placed in open, i.e., uncapped, and closed, i.e., capped vials. These vials were those used by the manufacturer to package and store the reagent test strips. One of each type of vial was placed in the manufacturer-recommended storage environment at room temperature and the adverse environments (incubator, direct light to mimic sunlight exposure, humidity, and refrigerated). The Accu-Chek Easy glucometer and reagent test strips as well as Accu-Chek Easy high and low glucose control solutions, manufactured by Roche, were used for this study. MAIN OUTCOME MEASURES: On day-3, day-7, and then once every 7 days, one strip from each vial in each environment was tested with the same glucometer using both a high and a low glucose control. The strip was considered failed for a type of vial and storage environment when either control was out of the reference range on a regular testing day and still out of range when tested the subsequent day. Testing continued up to 50 days. RESULTS: For the tested environments it was found that, overall, test strip stability lasted longer for closed vials than open vials. For open vials in adverse storage conditions, the refrigerator environment offered the longest stability at 35 to 50 days and direct light and humidity offered the shortest periods of stability at 3 to 14 days. CONCLUSIONS: The results of this study support the manufacturer's recommendations to store POCT glucose test strips in their original vial, capped, and at room temperature, though refrigeration may offer an alternative storage environment with acceptable stability. As compliance with testing, quality control, and storage instructions is often an issue with POCT, the manufacturers of these systems for blood glucose measurement should design storage systems that allow the patient to store the glucose meter and the reagent strips in the same location. Manufacturers may also need to consider designing storage systems that are more portable, knowing that patients must take the glucose meters and test strips with them when they travel. Roche's Accu-Chek Compact system is an example of such a design. The glucose test strips are incorporated into a drum that is stored in the Accu-Chek meter itself. When a patient performs a fingerstick blood glucose measurement, the drum advances to move a test strip outside the meter. When the test is complete, the test strip is ejected for disposal. Future studies to clarify the effect of adverse storage conditions, particularly refrigeration, on the integrity of POCT test systems and reagent strips is warranted with currently marketed brands. PMID- 16315737 TI - Improving the accuracy of specimen labeling. AB - Accurate specimen identification is a challenge in all hospitals. A mislabeled specimen can lead to devastating consequences for a patient. In an effort to decrease the risk of potential harm caused by labeling errors, Children's Hospitals and Clinics of Minnesota successfully implemented a Zero Tolerance Laboratory Specimen Labeling process. After months of studying, charting, networking, and communicating with all stakeholders the new process led to a 75% reduction in laboratory specimen labeling errors. PMID- 16315738 TI - Piloting case-based instruction in a didactic clinical immunology course. AB - OBJECTIVES: To assess (1) the effect of case-based instructional modules on student critical thinking, class attendance, and satisfaction and (2) student opinion of case formats. DESIGN/SETTING/PARTICIPANTS: University-based upper division course in clinical immunology and serology. The course was taught by the same instructor for two consecutive semesters with the intervention introduced in the second semester. Sixty-seven students experienced the intervention and 56 students were in the baseline cohort. INTERVENTION: Nine cases were interspersed between lectures during the semester. Each case took one 50-minute class in which students worked in groups of five or six. MAIN OUTCOME MEASURES: Student performance on five critical thinking multiple-choice examination questions and percent student attendance on case days versus lecture days were analyzed using the Mann-Whitney test. Student ratings on course evaluations were analyzed using t-test comparing semesters with and without intervention. Student opinion of cases was obtained through surveys and a focus group. RESULTS: Student performance on critical thinking exam questions was similar in the two groups. Student attendance was significantly higher on case days (95.6%) versus lecture days (80.3%; p < 0.0001). Only composite ratings for instructor involvement, student-instructor interaction, and course organization were significantly improved in the semester with cases compared to the semester with lecture only (p < 0.0001 for all indices). CONCLUSIONS: Although case studies did not significantly improve student performance on critical thinking questions, they still proved to be a valuable instructional method. Student attendance, student instructor interaction, and instructor involvement in the course were all positively affected by incorporation of case studies. Discussion of cases also helped to uncover student misconceptions of course material. PMID- 16315739 TI - Screening for diabetes: sensitivity and positive predictive value of risk factor total. AB - OBJECTIVE: Screening for diabetes is recommended for individuals > or =45 years of age, or earlier if they manifest > or = one specific risk factors. This study examined the sensitivity and positive predictive value (PPV) of risk factor total for identifying individuals with diabetes and prediabetes. DESIGN: Subjects were interviewed to assess the presence of risk factors. Fasting plasma glucose levels were obtained. SETTING: The study occurred at a health fair in Greensburg, PA. PATIENTS: Six hundred sixty-one Caucasians between the ages of 19 and 100. RESULTS: Using the criterion of screening individuals with > or = one risk factors detected 100% of both diabetics and prediabetics. This dropped to 91.2% when screening individuals with > or = two factors. The PPV of the risk factor total was poor (80% of individuals with a total of four factors were not diabetic). The ability of the risk factor total to predict individuals with impaired glucose metabolism (prediabetics + diabetics) was considerably better, and increased almost linearly with the risk factor total. Of the subjects with normal glucose values, the mean glucose increased as the risk factor total increased. CONCLUSION: While the sensitivity of using > one risk factor as an algorithm to screen is 100% for identifying diabetics, the PPV of risk factor analysis for identifying diabetics is poor. The same algorithm works well to identify at-risk individuals, presumably allowing early intervention and education. PMID- 16315740 TI - Students' perceptions of laboratory science careers: changing ideas with an education module. AB - OBJECTIVE: To assess the effectiveness of a Web-based education module in changing students' perceptions of laboratory science careers. DESIGN: Perception was measured with a short examination and then a Web-based exercise was presented. Following the exercise, the test was administered again. Frequency data from the pre-test and post-test were compared for changes in perception. The correlated pre-test/post-test pairs were also examined for opinion changes and these were analyzed for significance. SETTING: Large parochial high schools in New Orleans, Louisiana. A small team visited the schools during their appointed class times for biology. PARTICIPANTS: Study participants were high school biology students in grades 9-10. Two-hundred-forty-five students participated (149 male and 96 female). INTERVENTIONS: A Web-based exercise on blood film examination was presented to the students in a classroom setting (www.mclno.org/labpartners/index_03.htm). The exercise contained focused messages about: (1) the numbers of healthcare workers acquiring AIDS from on-the-job exposure and (2) common career paths available to the laboratory science workforce. MAIN OUTCOME MEASURES: The shift in perception of: What medical service generates the most diagnostic data. Which professional group performs laboratory tests. The risk of acquiring AIDS while working in the healthcare setting. Interest in a science-related career. How much education is required to work in a science-related field. RESULTS: The intervention significantly shifted perception in all areas measured except that of interest in a science-related career. CONCLUSIONS: Many students perceive that the risk of acquiring AIDS while working in the healthcare setting is "high". Web-based presentations and similar partnerships with science teachers can change perceptions that might lead to increased interest in clinical laboratory science careers. PMID- 16315741 TI - The reemergence of pertussis in immunized populations: a case study. AB - OBJECTIVE: To present a case of classical pertussis occurring in previously vaccinated male siblings, 11 and 13 years of age, living in El Paso TX; also to present an overview and update of the changing epidemiology of pertussis including pathophysiology, diagnosis, and treatment. DESIGN: Nasopharyngeal swabs and blood samples were collected from two male siblings, 11 and 13 years of age, presenting with cold-like symptoms and persistent cough during the second week of infection. Nasopharyngeal swabs were plated onto Bordet-Gengou agar plates and incubated for 48 hours. Blood samples were analyzed for the presence of antibodies (IgM and IgA) against Bordetella pertussis antigens using indirect enzyme linked immunosorbent assay (ELISA). SETTING: Cultures and serological analysis was conducted at the University of Texas at El Paso, Clinical Laboratory Science Program Research facility. RESULTS: Bacterial cultures of both children were positive for Bordetella pertussis and the sera revealed positive IgM and IgA antibodies (> 11 PANBIO UNITS) against a mixture of antigens including: pertussis toxin, filamentous hemagglutinin, pertactin, and fimbriae. CONCLUSIONS: Pertussis immunity wanes overtime, leaving most adolescents and adults susceptible to infection. Physicians must be prepared to diagnose and treat pertussis in any age group regardless of vaccination status. PMID- 16315742 TI - Laboratory managers' views on attrition and retention of laboratory personnel. AB - OBJECTIVE: This study was undertaken to provide information on the current shortage of clinical laboratory employees and to identify strategies for retaining laboratory employees. DESIGN: A paper survey was distributed to 800 clinical laboratory managers. SETTING: The survey was sent to laboratory managers at their work sites. PATIENTS OR OTHER PARTICIPANTS: 190 usable surveys were returned for a response rate of 24%. INTERVENTIONS: Surveys were mailed in March 2003. MAIN OUTCOME MEASURES: The number of CLTs and CLSs considered fully staffed at the laboratory managers' institutions, the numbers of CLTs and CLSs who left the institutions in a five-year period, and the reasons employees left were tabulated. The managers' responses to questions on the factors that they considered most important in retaining laboratory employees were tabulated and categorized. RESULTS: In this five-year period (1998-2002), 5% of employees left their jobs annually. Over 60% of laboratory employees who left did so in the first five years of practice. The top five reasons that employees left their jobs were: (1) new laboratory job, (2) moved/family obligations, (3) retirement, (4) left the field entirely, and (5) employee was fired. In the first year of practice, 15% of the employees who left were fired. Between one and five years of practice, 7.3% left because of the hours or shift, 6.7% left to pursue further education for a non-laboratory career, and 6.7% left the field entirely. In the group of employees who left between five and ten years, 13.5% left the field entirely and 5.2% left for sales or clinical trials positions. Over 40% of the employees with more than ten years of experience who left did so because of retirement. CONCLUSION: Most laboratory employees who left did so to take another laboratory position; however, reasons for leaving vary with years of experience. The number of laboratory employees leaving the profession exceeds the number of new graduates entering the profession making the retention of employees essential. Laboratory managers identified salary as the most important retention factor. PMID- 16315743 TI - A qualitative assessment of systematic instructional design training by CLS faculty members. AB - OBJECTIVE: To determine the perceived value clinical laboratory science (CLS) faculty members gave to their participation in workshops on the use of a modified systematic instruction design (SID) model to develop curriculum and on-line courses. DESIGN: A survey assessing the perceived value of SID training was sent to 27 CLS faculty members. The survey asked the respondents to assess the value of the training that they received in developing their skills in Web-based, distance learning course development and teaching, and expanding their skills in traditional course development and teaching. The eight components of SID were listed and the respondents rated each component as to its value to them on a 5 point Likert scale of 5 = very valuable to 1 = not very valuable. In addition to rating the value of each SID component, the respondents were asked if they would like more training in any of the eight components. RESULTS: A majority of the 18 respondents (67%) reported that the training in SID was valuable to them. A strong majority of the respondents indicated that their training in goal and instructional analyses (96%), media selection (94%), and aligning objectives, assessments, and instructional strategies (94%) were valuable to their distance education programs and their traditional teaching skills. CONCLUSION: Faculty members who actively participated in SID training valued their new skills in developing distance education courses as well as improving their traditional teaching activities. Research is needed on the effect these new teaching skills have on student learning. PMID- 16315744 TI - From single cell gene-based diagnostics to diagnostic genomics: current applications and future perspectives. AB - Molecular diagnostics is a branch of clinical diagnostics that uses primarily DNA or RNA as a biomarker for clinical testing. It combines various gene-based amplification technologies with highly sophisticated detection methods for the clinical diagnosis of a vast variety of diseases including infectious diseases, cancer, and inherited diseases. The principal application of gene-based amplification technology is to identify pathogen or gene-specific nucleic acid sequences that are used as surrogate markers for the identification of either infectious pathogens or alteration of disease-related genes. There are generally three classes of gene-based amplification technologies: target-based, e.g., PCR; probe-based, e.g., LCR; and signal-based, e.g., bDNA. Real-time detection of PCR allows us to quantify amplified amplicons with a broad dynamic range and it offers a unique way to detect genetic mutations. Other technologies such as immuno-PCR and bio-barcode assay (BCA) combine different amplification tactics offering extreme detection sensitivity ranging from femtogram (10(-15)) to zeptogram (10(-21)). Even though quantum dots technology is in its infant stage, its potential to further increase diagnostic sensitivity and specificity is likely beyond our current imagination. Future diagnostic technologies include the use of genomic and proteomic approaches especially in pure cell types or even in the single-cell level, which open up endless new possibilities for gene-based diagnostics at entirely different levels. In this article, principles of various current gene-based amplification and detection technologies along with their clinical applications are discussed. New technologies that could potentially be used in future gene-based diagnosis are introduced. PMID- 16315745 TI - Molecular-based laboratory testing and monitoring for human immunodeficiency virus infections. AB - Applications of laboratory testing for human immunodeficiency virus type 1 (HIV 1) infection have made significant impact on clinical care of HIV-infected patients globally. As these technologies continue to evolve and new technologies emerge, unique and highly sensitive nucleic acid-based testing methods will offer more and better means for us to guide physicians in anti-retroviral treatment strategies and clinical management of HIV infected patients. In this review we discuss a variety of current molecular-based methods that are available for HIV testing including diagnosis, monitoring disease progression, and detection of drug resistance to anti-retroviral therapy. Newer approaches that could be used in future HIV testing are also introduced. PMID- 16315746 TI - Molecular diagnostics of inherited thrombosis. AB - Thrombophilia can best be defined as a disorder of coagulation that contributes to a predisposition towards thrombosis. Although the term thrombophilia has been used to describe arterial thrombosis, its most common usage has been in reference to venous thromboembolism (VTE). Thrombophilia can be a consequence of both acquired and inherited or genetic causes. Acquired causes include conditions such as surgery, cancer, and prolonged immobilization, while genetic causes have been linked to the inherited deficiencies of antithrombin, protein C, and protein S. The identification of the genetic basis of these inherited causes of thrombophilia ushered in a new way of thinking about thrombosis and the importance of its genetic component. Interest in the genetic basis of VTE was accelerated with the subsequent discovery of factor V Leiden, prothrombin G20210A, and MTHFR C677T. These single nucleotide polymorphisms (SNPs) and other genetic variants associated with VTE have become fixtures in the molecular diagnosis of inherited thrombophilia. Because of the large volume of current and anticipated future genetic testing, there has been a push to develop many different genotyping methods which are now used in both clinical and research settings. The identification of new genetic variants that may either directly or indirectly affect coagulation or the anticoagulant pathway, may greatly advance the understanding and clinical management of thrombophilia. PMID- 16315747 TI - [Changing patterns of communicable diseases in Korea]. AB - Before twentieth centuries and during early twentieth centuries, communicable diseases were the major cause of morbidity and mortality in Korea. But reliable data are not available. After 1975, the overall morbidity and mortality from communicable diseases, rapidly declined. Recently many new pathogenic microbes were recognized: L. monocytogenes, Hantaan virus, Y. pseudotuberculosis, P. multocida, L. pneumophilia, Human immunodeficiency virus (HIV), G. seoi, H. capsulatum, C. burnetii, V. cholerae 0139, C. parvum, F. tularensis, E. coli 0157:H7, B. burgdorferi, S. Typhimurium DT104, Rotavirus, hepatitis C virus and so on. Since the first HIV infection recognized in 1985, the reported cases of infection and deaths from HIV/AIDS have been steady increased each year. Legionnaire's disease, E. coli 0157:H7 colitis, listeriosis and crytosporidiasis have been occurring just sporadically among immunocompromized cases. Many re emerging communicable diseases were occurred in Korea: leptospirosis, malaria, endemic typhus, cholera, tsutsugamushi disease, salmonellosis, hepatitis A, shigellosis, mumps, measles, acute hemorrhagic conjunctivitis, brucellosis and so on. Leptospirosis and tsutsugamushi diseases have been noticed as major public health problems since 1980s. The malaria that had been virtually disappeared for a decade has reappeared from 1993 with striking increase of patients in recent 3 4 years. The distributions of salmonella and shigella serotypes have been changed a lot in recent few decades. Furthermore rapid emergence of antibiotic-resistant bacterial strains induces more difficult and complex problems in control of communicable diseases. We must recognize on the importance of environment and ecosystem conservation and careful prescription of anti-microbial agent in order to prevent communicable diseases. PMID- 16315748 TI - [Epidemiology and control of injury]. AB - Injury has recently become a major world-wide health problem. Injury related deaths occur in many actively working young people and produce major social and economical losses. However health related specialists do not recognize the importance of injury and there have not been many studies related to this problem. This research studied the trends of injury related research in Korea, mortality rate and prevalence rate, socio-economical losses and control in Korea and out of the country, based on literature from Korea and without such as statistical yearly reports on causes of deaths and OECD health reports, as well as WHO web sites. Studies in Korea about injury were very few, with 9 in the 1960's, 5 in the 1980's, 4 in the 1990's and 5 in 2000's. Mortality rate of injury was higher in Korea than in England, USA or Japan, especially in car accidents, suicide and falls. In Korea, the yearly trends in mortality rates were highest in car accidents but those rates are falling, suicide is steadily rising, with highest rate in 2003. Falls is in second rank with no change in rates. In 2003, the ten causes of death in Korea were suicide in 5th rank, transport accidents in 7th rank, and falls in 10th rank. Considering age groups, in the teens, transport accidents were 1st rank, in the 20's and 30's, suicide was 1st rank, and although there were some differences, falls, drowning, assault, fire were in the top 10. Prevalence rates of injury could not be known, but in 2001, according to the National Health and Nutrition Survey, lifelong injury was 10%, and yearly major injury was 1.3%, major injury for two weeks was 0.1%, and minor injury was 10%. In other foreign countries, injury has become to be recognized as a major health related problem, and much programs are being set up to reduce injury related deaths and injuries. WHO is putting much effort in prevention of violence and transport accidents, and in the USA, Canada and Europe, there are injury surveillance systems. Recently, as suicide is increasing rapidly and providing much problems, each country are managing suicide prevention programs. In Korea, Ministry of Construction and Transportation is managing and guiding the policies for prevention of transport accidents. In September of 2004, the Ministry of Health and Welfare has set up a 5 year plan of suicide prevention. PMID- 16315749 TI - [Impact of risk adjustment with insurance claims data on cesarean delivery rates of healthcare organizations in Korea]. AB - OBJECTIVES: To propose a risk-adjustment model from insurance claims data, and analyze the changes in cesarean section rates of healthcare organizations after adjusting for risk distribution. METHODS: The study sample included delivery claims data from January to September, 2003. A risk-adjustment model was built using the 1st quarter data, and the 2nd and 3rd quarter data were used for a validation test. Patients' risk factors were adjusted using a logistic regression analysis. The c-statistic and Hosmer-Lemeshow test were used to evaluate the performance of the risk-adjustment model. Crude, predicted and risk-adjusted rates were calculated, and compared to analyze the effects of the adjustment. RESULTS: Nine risk factors (malpresentation, eclampsia, malignancy, multiple pregnancies, problems in the placenta, previous Cesarean section, older mothers, bleeding and diabetes) were included in the final risk-adjustment model, and were found to have statistically significant effects on the mode of delivery. The c statistic (0.78) and Hosmer-Lemeshow test (chi2=0.60, p=0.439) indicated a good model performance. After applying the 2nd and 3rd quarter data to the model, there were no differences in the c-statistic and Hosmer-Lemeshow chi2. Also, risk factor adjustment led to changes in the ranking of hospital Cesarean section rates, especially in tertiary and general hospitals. CONCLUSION: This study showed a model performance, using medical record abstracted data, was comparable to the results of previous studies. Insurance claims data can be used for identifying areas where risk factors should be adjusted. The changes in the ranking of hospital Cesarean section rates implied that crude rates can mislead people and therefore, the risk should be adjusted before the rates are released to the public. The proposed risk-adjustment model can be applied for the fair comparisons of the rates between hospitals. PMID- 16315750 TI - [Field study of concentrations and emissions of particulate contaminants by types of swine houses in Korea]. AB - OBJECTIVES: Particulate contaminants, such as total and respirable dusts, can harm the health of farm workers via several routes. The principal aims of this field study were to determine the concentrations and emissions of particulate contaminants: total and respirable dusts, in the different types of swine houses used in Korea, and allow objective comparison between Korea and the other countries in terms of swine housing types. METHODS: The swine houses investigated in this research were selected with respect to three criteria: the manure removal system, ventilation mode and growth stage of pigs. Measurements of total and respirable dust concentrations and emissions in the swine houses were carried out on 5 housing types at 15 different farm sites per housing type. The swine houses investigated were randomly selected from farms situated within the central districts in Korea: province of Kyung-gi, Chung-buk and Chung-nam. RESULTS: The total and respirable dust concentrations in the swine houses averaged 1.88 and 0.64 mg/m3, ranging from 0.53 to 4.37 mg/m3 and from 0.18 to 1.68 mg/m3, respectively. The highest concentrations of total and respirable dusts were found in the swine houses with deep-litter bed systems: 2.94 mg/m3 and 1.14 mg/m3, while the lowest concentrations were found in the naturally ventilated buildings with slats: 0.83 mg/m3 and 0.24 mg/m3, respectively (p<0,05). All the swine houses investigated did not exceed the threshold limit values (TLVs) for total (10 mg/m3) and respirable (2.5 mg/m3) dusts. The mean emissions of total and respirable dusts, per pig (75 kg in terms of live weight) and area (m2), from the swine houses were 97.33 and 9.55 mg/h/pig and 37.14 and 12.83 mg/h/ m2, respectively. The swine houses with deep-litter bed systems showed the highest emissions of total and respirable dusts (p<0.05). However, the emissions of total and respirable dusts from the other swine houses were not significantly different (p>0.05). CONCLUSION: The concentrations and emissions of total and respirable dusts were relatively higher in the swine houses managed with deep-litter bed systems and ventilated naturally of the different swine housing types tested. In further research, more farms than the number used in this research should be investigated, which will present objective and accurate data on the concentrations and emissions of total and respirable dusts in Korean swine houses. In addition, personal sampling should be performed to objectively assess the exposure level of farm workers to particulate contaminants. PMID- 16315751 TI - [Census population vs. registration population: which population denominator should be used to calculate geographical mortality]. AB - OBJECTIVES: Studies on the geographical differences in mortality tend to use a census population, rather than a registration population, as the denominator of mortality rates in South Korea. However, an administratively determined registration population would be the logical denominator, as the geographical areas for death certificates (numerator) have been determined by the administratively registered residence of the deceased, rather than the actual residence at the time of death. The purpose of this study was to examine the differences in the total number of a district population, and the associated district-specific mortality indicators, when two different measures as a population denominator (census and registration) were used. METHODS: Population denominators were obtained from census and registration population data, and the numbers of deaths (numerators) were calculated from raw death certificate data. Sex- and 5-year age-specific numbers for the populations and deaths were used to compute sex- and age-standardized mortality rates (by direct standardization methods) and standardized mortality ratios (by indirect standardization methods). Bland-Altman tests were used to compare district populations and district specific mortality indicators according to the two different population denominators. RESULTS: In 1995, 9 of 232 (3.9%) districts were not included in the 95% confidence interval (CI) of the population differences. A total of 8 (3.4%) among 234 districts had large differences between their census and registration populations in 2000, which exceeded the 95% CI of the population differences. Most districts (13 of 17) exceeding the 95% CI were rural. The results of the sex- and age-standardized mortality rates showed 15 (6.5%) and 16 (6.8%) districts in 1995 and 2000, respectively, were not included in the 95% CI of the differences in their rates. In addition, the differences in the standardized mortality ratios using the two different population denominators were significantly greater among 14 districts in 1995 and 11 districts in 2002 than the 95% CI. Geographical variations in the mortality indicators, using a registration population, were greater than when using a census population. CONCLUSION: The use of census population denominators may provide biased geographical mortality indicators. The geographical mortality rates when using registration population denominators are logical, but do not necessarily represent the exact mortality rate of a certain district. The removal of districts with large differences between their census and registration populations or associated mortality indicators should be considered to monitor geographical mortality rates in South Korea. PMID- 16315752 TI - [The relations of socioeconomic status to health status, health behaviors in the elderly]. AB - OBJECTIVES: To analyze the relationships of socioeconomic status(SES) to health status and health behaviors in the elderly. METHODS: Data were obtained from self administered questionnaire of 4,587 persons, older than 65 years, living in a community. We measured the sociodemographic characteristics, socioeconomic status, health status (subjective health status, acute disease, admission experience, dental state, chronic disease etc.), activities of daily living (ADL), instrumental activities of daily living (IADL), and mini-mental state examination-Korean (MMSE-K). Binary and multinominal logistic regression analyses were employed to analyze factors affecting on the socioeconomic status of the elderly. RESULTS: With regard to the SES and health status, those with a low SES had poorer subjective health states and lower satisfaction about their physical health. Also, acute disease experiences, admission rates and tooth deciduation rates were higher in those of low SES. In the view of physical and cognitive functions, the ADL, IADL and MMSE-K scores were also lower in those of low SES. However, with regard to health behaviors, lower smoking and alcohol drinking rates were found in the low SES group, and a similar trend was shown with regular physical exercise, eating breakfast, and regular physical health check-up. From these findings, we surmise that those with low SES have a poorer health condition and less money to spend on health, therefore, they can not smoke or drink alcohol, exercise and or have a physical health check-up. CONCLUSION: This study suggests that socioeconomic status plays an important role in health behaviors and status of the elderly. Low socioeconomic status bring about unhealthy behavior and poor health status in the elderly. Therefore, more specific target oriented(esp. low SES persons) health promotion activities for the elderly are very important to improve not only their health status, but their health inequity also. PMID- 16315753 TI - [An epidemiological study for child pedestrian traffic injuries that occurred in school-zone]. AB - OBJECTIVES: Pedestrian traffic injuries have been an important cause of childhood mortality and morbidity for decades. The aim of this study was to describe the epidemiological characteristics of child pedestrian traffic injuries that occurred during 2000 in one metropolitan city and its school-zones, and to determine the factors associated with those accidents. METHODS: A cross-sectional study was performed in 2001. Police records were used to identify the cases of pedestrian injury. Children aged between 6 and 15 years, injured during road walking, were included in this study. A direct survey of the environmental factors within the school-zones in study area (n=116) was also performed. Self administered questionnaires, via mail and telephone surveys, were used to assess the safety education programmes. The schools were divided into two groups according to the occurrence of pedestrian traffic injuries in their school-zone. RESULTS: Pedestrian injuries (n=597) were found to account for 3.2% of all traffic injuries in the subject area. The epidemiological characteristics were not significantly different between genders. There were some significant risk factors within the environmental factors, such as local road (OR: 2.3, 95% CI=1.05-5.35), heavy traffic volume (OR: 2.2, 95% CI=1.00-5.04), poor visibility of speed-limit signs (OR: 2.8, 95% CI=1.25-6.42), no separation of pedestrian routes from cars (OR: 2.6, 95% CI=1.02-6.75) and barriers on the pedestrian routes (OR: 2.2, 95% CI=1.01-5.08). Only one factor, that of education in a safety-park (OR: 0.3, 95% CI=0.09-0.96), was significantly associated in the traffic and pedestrian safety education factors. CONCLUSION: Significant associations with pedestrian injury risk were identified in some of the modifiable environmental factors than in the educational factors. PMID- 16315754 TI - [A study of immune response to hepatitis B vaccine & HBV DNA in isolated anti-HBc positive subjects]. AB - OBJECTIVES: The aim of this study was to evaluate the response to a hepatitis B vaccination, and investigate the HBV DNA in subjects with isolated anti-HBc. METHODS: 34 subjects with persistent isolated anti-HBc were included in the study. 32 subjects negative for HBsAg, anti-HBs and anti-HBc were included as a control group. They were all vaccinated with Hepaccine at 0, 1 and 2 months, and anti-HBs titers were measured 1 month after the 1st and 3rd vaccinations (1 and 3 months). The HBV-DNA was tested by polymerase chain reaction in subjects with isolated anti-HBc. RESULTS: After the 1st & 3rd vaccinations, the anti-HBs titers > or = 10 mlU/ml were 70.6 & 70.6% in isolated anti-HBc group, and 34.4 & 81.2% in the control group, respectively. There were statistically significant differences after the 1st vaccination, but none after the 3rd, between the two groups. In the isolated anti-HBc and control groups, the primary, amnestic and no responses were 0 vs. 46.9%, 55.9 vs. 6.3% and 29.4 vs. 18.8%, respectively. The HBV DNA was not detected in all subjects with isolated anti-HBc. CONCLUSION: None of the subjects with isolated anti-HBc had a false positive result (primary response); therefore, they should be excluded from vaccination programs in Korea. To differentiate between immunity and occult infections, a single dose of vaccine, with a follow-up anti-HBs test, is preferable for subjects with isolated anti-HBc. An amnestic response indicates late immunity, and no response a suspect occult infection. PMID- 16315755 TI - [Estimating the burden of diseases due to high alcohol consumption in Korea]. AB - OBJECTIVES: This study estimated the burden of disease due to high alcohol consumption using DALY, a composite indicator recently developed by the Global Burden of Disease study group. The results were analyzed by age and sex. METHODS: Firstly, high alcohol consumption-related diseases, and their relative risk (RR), were selected. Secondly, population attributable fractions (PAFs) were computed using formulae, including the relative risk (RR) and prevalence of exposure (Pe). Thirdly, the DALYs of high alcohol consumption-related diseases were estimated. Lastly, the attributable burdens of diseases due to high alcohol consumption wereconcluded as being the sum of the products that multiplied the DALYs of high alcohol consumption-related diseases by their population attributable fraction (PAF). RESULTS: The burden of high alcohol consumption in Korea was 2992.3 person years (PYs) per 100,000 persons in men, and 1426.6 in women. For men, the high alcohol consumption-induced diseases with the five biggest burdens were liver cirrhosis, hypertensive disease, liver cancer, cerebral infarction and intracerebral hemorrhage. For women, these were cerebral infarction, intracerebral hemorrhage, hypertensive disease, liver cirrhosis and liver cancer. CONCLUSION: This study highlighted the attributable fraction of diseases due to exposure to high alcohol consumption, by quantifying the results of exposure to risk factors. Therefore, it is now possible to assess interventions for risk factors in quantifiable terms in each population. Finally, measuring the risk factor burdens was expected to contribute to priority setting and effective resource allocation in public health policy. PMID- 16315756 TI - [Epidemiologic investigation on sporadic occurrence of shigellosis in a subcounty of Cheongwon County in Chungbuk Province in 2003]. AB - OBJECTIVES: This study was undertaken to investigate the source of infection and mode of transmission of shigellosis, which occurred sporadically among residents and students in a subcounty of Cheongwon county, Chungbuk province, Korea, from June 4 to July 3 2003. METHODS: 692 subjects completed a questionnaire and provided a swab for microbiological examinations and 7 environmental specimens were examined for bacterial organisms. PFGE (pulsed-field gel electrophoresis) and fingerprinting were performed to find the genetic relationship among the temporally associated sporadic isolates. RESULTS: A total of 29 patients had symptoms consistent with the case definition, with 13 confirmed and 16 suspected cases. The frequency of diarrhea was 6 times or more a day (80.8%), with a duration of 1 to 4 days (88.5%) in most cases. The most common symptoms accompanying the diarrhea were fever (80.9%) followed by abdominal pain (76.9%), headache (65.4%), chill (61.5%), vomiting (46.2%) and tenesmus (15.4%). The epidemic curve was characteristic of a person-to-person transmission. The PFGE and fingerprinting demonstrated identical or similar DNA patterns among the 3 Shigella sonnei isolates (A51, A53 and A61 types) found in this outbreak. CONCLUSION: A genetically identical strain of S. sonnei was estimated to be the cause of this outbreak, and the mode of transmission was most likely person-to person. PMID- 16315757 TI - [Epidemiologic investigation on an outbreak of shigellosis in Seongju-gun, Korea, 2003]. AB - OBJECTIVES: An outbreak of shigellosis occurred among students and staff of S primary and middle school, Seongju-gun, in 2003. This investigation was carried out to institute an effective counterplan, and study the infection source and transmission of the shigellosis. METHODS: The authors conducted a questionnaire survey among 235 students and staff from S preschool, primary and middle school relating to the ingestion of school lunch and the manifestation of symptoms. Also, the author investigated the drinking water, feeding facility and reconstructed cooking process of the food presumed to be the cause of the shigellosis. The diarrhea cases were defined as confirmed cases and those cases who had had diarrhea more than one time, accompanied with symptoms such as fever, vomiting and tenesmus. RESULTS: From rectal swabs 20 people, between June 28 and July 4, 2003, were confirmed with shigellosis. The diarrhea attack rate was 40.0%. Those who had ingested tomatoes and cubed radish kimchi had significantly higher diarrhea attack rates (p<0.05), with the relative risk of tomatoes being 2.69 (95% CI: 0.98-7.42). The major cause of shigellosis was presumed to be from contaminated tomatoes due to cooking with rubber gloves containing holes. CONCLUSION: The cooks in charge of school lunches must make doubly sure to not only attend to their sanitation, but also to manage the table wear and items used in providing school lunches. The health care authority should introduce higher leveled criteria for health care among cooks, so that they cannot cook when the have a case of any infectious disease. PMID- 16315758 TI - [Effects of the severe asian dust events on daily mortality during the spring of 2002, in Seoul, Korea]. PMID- 16315759 TI - [What will be the proper criteria for impaired fasting glucose for Korean men? Based on medical screening data from a general hospital]. AB - OBJECTIVES: Recently, the American Diabetes Association (ADA) redefined the criteria of prediabetes, which has lowered the diagnostic level of fasting plasma glucose (FPG) from 110 to 125 mg/dl, down to levels between 100 to 125mg/dl. The purpose of this study was to determine the predictive cutoff level of FPG as a risk for the development of diabetes mellitus in Korean men. METHODS: A retrospective cohort study was conducted on 11,423 (64.5%) out of 17,696 males < or =30 years of age, and who met the FPG of < or = 125 mg/dl and hemoglobin A1c of < or = 6.4% criteria, without a history of diabetes, and who were enrolled at the screening center of a certain university hospital between January and December 1999. The subjects were followed from January 1999 to December 2002 (mean follow-up duration; 2.3(+/-0.7) years). They were classified as normal (FPG <100mg/dl), high glucose (FPG > or =100 mg/dl and < 110 mg/dl) and impaired fasting glucose (FPG 2110 mg/dl and < 125 mg/dl) on the basis of their fasting plasma glucose level measured in 1999. We compared the incidence of diabetes between the 3 groups by performing Cox proportional hazards model and used receiver operating characteristic analyses of the FPG level, in order to estimate the optimal cut-off values as predictors of incident diabetes. RESULTS: At the baseline, most of the study subjects were in age in their 30s to 40s (mean age, 41.8(+/-7.1) year). The incidence of diabetes mellitus in this study was 1.19 per 1,000 person-years (95% CI=0.68-1.79), which was much lower than the results of a community-based study that was 5.01 per 1,000 person-years. The relative risks of incident diabetes in the high glucose and impaired fasting glucose groups, compared with the normal glucose group, were 10.3 (95% CI=2.58-41.2) and 95.2 (95% CI= 29.3-309.1), respectively. After adjustment for age, body mass index, and log triglyceride, a FPG greater than 100 mg/dl remained significant predictors of incident diabetes. Using the receiver operating characteristic (ROC) curve, the optimal cutoff level of FPG as a predictor of incident diabetes was 97.5 mg/dl, with a sensitivity and a specificity of 81.0% and 86.0%, respectively. CONCLUSION: These results suggest that lowering the criteria of impaired fasting glucose is needed in Korean male adults. Future studies on community-based populations, including women, will be required to determine the optimal cutoff level of FPG as a predictor of incident diabetes. PMID- 16315760 TI - [Factors affecting re-smoking in male workers]. AB - OBJECTIVES: This study was performed to examine the factors affecting re-smoking in male workers. METHODS: A self-administrated questionnaire survey was conducted during April 2003 to examine the smoking state of 1,154 employees of a company that launched a smoking cessation campaign in 1998. Five hundred and eighty seven persons, who had stopped smoking for at least one week, were selected as the final study subjects. This study collected data on smoking cessation success or failure for 6 months, and looked at the factors having an effect on resmoking within this period. This study employed the Health Belief Model as its theoretical basis. RESULTS: The re-smoking rate of the 587 study subjects who had stopped smoking for at least one week was 44.8% within the 6 month period. In a simple analysis, the resmoking rates were higher in workers with a low age, on day and night shifts, blue collar, of a low rank, where this was their second attempt at smoking cessation and for those with a shorter job duration (p<0.05). Of the cues to action variables in the Heath Belief Model, re-smoking was significantly related with the perceived susceptibility factor, economic advantages of smoking cessation among the perceived benefits factor, the degree of cessation trial's barrier of the perceived barriers factor, smoking symptom experience, recognition of the degree of harmfulness of environmental tobacco smoke and the existence of chronic disease due to smoking (p<0.05). In the multiple logistic regression analysis for re-smoking, the significant variables were age, perceived susceptibility for disease, economic advantages due to smoking cessation, the perceived barrier for smoking cessation, recognition on the degree of harmfulness of environmental tobacco smoke, the existence of chronic disease due to smoking and the number of attempts at smoking cessation (p<0.05). CONCLUSION: From the result of this study, for an effective smoking ban policy within the work place, health education that improves the knowledge of the adverse health effects of smoking and the harmfulness of environmental tobacco smoke will be required, as well as counter plans to reduce the barriers for smoking cessation. PMID- 16315761 TI - Physiology and pathology of NMDA receptors. AB - Ionotropic glutamate receptors of the N-methyl-D-aspartate (NMDA) subtype are highly expressed in the central nervous system and are involved in excitatory synaptic transmission and synaptic plasticity. Prolonged activation of NMDA receptors can lead to excitotoxicity, which is implicated in the pathogenesis of neurodegeneration occurring in various acute and chronic disorders of the central nervous system. Recent advances in understanding the function, pharmacology, genetics and structure of NMDA receptors has promoted a search for new compounds that could be therapeutically used. These compounds act on agonist binding sites, either apart from them or directly within the ion channel pore. Members of the last group are called open channel blockers, and some of them, such as memantine and ketamine, are already clinically used. Kinetic modeling of NMDA receptor activity was employed to define the effects of various groups of modulators. Quantifying the action of these substances by kinetic parameters can help us to reveal the molecular mechanism of action at the receptor and to characterize the dependence of its action on the mode of NMDA receptor activation. Two modes are considered: phasic activation, induced by synaptically released glutamate, and tonic activation, which is expected to occur under pathological conditions when low, but sustained levels of glutamate activate NMDA receptors. The aim of our review is to summarize the recent data about the structural and functional properties of NMDA receptors and their role in long-term potentiation and excitotoxicity. PMID- 16315762 TI - The impact of diabetes mellitus on skeletal health: an established phenomenon with inestablished causes? AB - Diabetes mellitus and osteoporosis affect a large proportion of older adults. In this context, diabetes may influence the bone in multiple pathways, some with contradictory effects. These mechanisms include changes in insulin and insulin like growth factors levels, hypercalciuria associated with glycosuria, reduced renal function, obesity, higher concentrations of advanced glycation end products in collagen, angiopathies, neuropathies and inflammation. Although it is assumed that the decreased bone strength in diabetes may contribute to fracture risk, a very high number of available clinical and/or epidemiological studies as well as animal model studies brought about heterogeneous or even contradictory results on the skeletal involvement in patients with diabetes mellitus. In addition, bone mineral density (BMD) is a convenient predictor for fracture and the type 1 diabetes is associated with modest reductions in BMD. However, type 2 diabetes can be related to the elevated BMD. The immediate improvement in these discrepancies is to consider the complex pathophysiology of diabetes as well as influences of gender, age, treatment and duration of the disease. It is important also to improve further the choice of investigated biochemical markers and the standardization of the bone mass measurements. Along these lines, several recent cohort studies undeniably indicated that diabetes itself is associated with increased risk of osteoporosis. PMID- 16315763 TI - Changing of facial skeleton for treatment of obstructive sleep apnoea syndrome. AB - Obstructive sleep apnoea syndrome (OSAS) is a potentially life-threatening disorder. It is characterized by at least five episodes of apnoea or hypopnoea during sleep lasting for more than 10 seconds. Apnoea or hypopnoea are accompanied by respiratory efforts. Changes of the facial skeleton by mandibular or maxillo-mandibular advancement belong to surgical techniques which might affect moderate and severe OSAS. In the surgical procedure mandible alone or the upper and lower jaws are moved forward by at least 10 mm. Thus also muscles fixed to the facial skeleton and upper airway dilatators are moved forward. The discussion also mentions possible complications and limitations of this surgical technique. PMID- 16315764 TI - Combined pelvic osteotomy in the treatment of both deformed and dysplastic acetabulum three years--prospective study. AB - Kuwait has a very high incidence of all types of developmental dysplasia of the hip. One of the most complicated forms is the deformed dysplastic acetabulum, which has not only abnormal orientation but it also has lost its shape by the pressure of the subluxed femoral head. The outer part of the socket involves the upper part of the original acetabulum, thus giving it a bipartite appearance. Clear border like edge separates this outer part from the inner part, which represents the lower part of the original acetabulum and it has no direct contact with the femoral head. Combined pelvic osteotomy (CPO) restores both the original shape and treats the malalignment of the acetabulum. Twelve children, aged between 18 and 54 months underwent CPO, two of them bilaterally. The follow up is between 36 - 60 months. In all cases, concentric stable reduction with good acetabular cover was obtained. PMID- 16315765 TI - Advanced glycoxidation end products in patients with multiple sclerosis. AB - Advanced glycoxidation end products (AGEs) play an important role in the pathogenesis of neurodegenerations and we studied if AGEs could represent a useful marker in patients with multiple sclerosis (MS). AGE-products were assessed in cerebrospinal fluid (CSF) and serum of 31 patients with MS and 8 controls. We did not find any statistically significant differences in patients with MS and controls either in CSF or in serum. We have observed a significant association between pentosidine and total AGEs as well as a relationship of both to the protein content in CSF in MS patients. Despite of the involvement of both oxidative stress and RAGE (receptor for AGEs) in the pathogenesis of MS and its experimental model, neither pentosidine nor total AGE were shown as useful markers in this indication. Other compounds and ligands of RAGE are probably of higher significance in MS. PMID- 16315766 TI - Chromatographic analysis in bacteriologic diagnostics of blood cultures, exudates, and bronchoalveolar lavages. AB - This article summarizes our previously achieved and published results. The method for the determination of bacterial volatile fatty acid patterns (VFA) in clinical samples was elaborated. It employs gas chromatography (GC), solvent extraction or head-space solid phase microextraction (SPME). This method was validated by analyses of reference bacterial strains. After cultivation in defined media, aerobic and facultative anaerobic bacteria provided profiles with a low or none acid content, while anaerobic bacteria provided characteristic but medium dependent profiles with a higher acid content. This method was used for the analyses of clinical samples of total 375 blood cultures, 205 suppurative and apyogenous exudates, and 210 bronchoalveolar lavages (BALs). These analyses enabled within 30 minutes the detection of microbes, probably non-sporulating anaerobes not found by false-negative cultivation, in 11.2% of blood cultures, in 20.0% of exudates, and in 9.0 to 20.0% of BALs. Using the mass spectrometry (MS) methods, a number of other components with unclear diagnostic importance were found in BAL samples, in particular hydrogen cyanide, methanol, ethanol, hexanol, acetone, cyclohexanone, acetonitrile, formaldehyde, acetaldehyde, ethyl acetate, and other esters. Cyclohexanone, occurring mainly in BALs of patients with pneumonia, undergoing intensive care, may originate as a residual solvent from the plastic parts of the ventilation apparatus. PMID- 16315767 TI - Smoking and pregnancy: prevalence, knowledge, anthropometry, risk communication. AB - Information about smoking impact on pregnancy as well as clear advice to stop and a treatment offer should be included into the health care of pregnant women. We contacted them within the first 3 days after delivery. In the sample of 265 women 23.8% (63/265) smoked during their pregnancy (51.9% with basic education only, 25.8% with high school and 5.0% with university education), and 7% of the sample (19/265) did not quit by the delivery. Only 68% of smokers (43/63) were asked about their smoking habits during the pregnancy by their gynaecologist, and both smokers and non-smokers had insufficient information about the impact of smoking on their baby (40% of smokers and 32% of non-smokers had no idea at all), the doctor was the source of this kind of information only in 5% (13/265) cases. Smokers' children had a lower average birth weight (3.084 g in smokers vs. 3.325 g in non-smokers, p=0.02) and were smaller (49.3 cm vs. 50.5 cm respectively, p=0.02). Smokers' bodyweight increased more than non-smokers' during their pregnancy (gaining 14.8 kg vs. 12.9 kg respectively, p=0.02), they also underwent the delivery at a lower age (27.6 vs. 30.0 years respectively, p<0.01) and after a non-significantly shorter duration of pregnancy (38.7 weeks vs. 39.0 weeks respectively, p=0.53). If 23.8% of pregnant women smoke and 90.6% of them would like to stop, it is a missed opportunity for education and treatment of tobacco dependence by their physicians. PMID- 16315768 TI - GIST of pancreas. AB - Authors present the case (from the year 2003) of a 38 years old patient, female with large tumor in the abdominal cavity. Preoperative differential diagnostic examination showed either tumor of pancreas or left liver lobe. The surgery confirmed large tumor body of the pancreas. Histological and immunohistochemical examination determined gastrointestinal stromal tumor (GIST) of the pancreas from autonomous nerves with malignant character. Tumor was radically removed. In the postoperative period regular follow up was done at the department of oncology, abdominal ultrasound every 3 months, CT scan every 6 month. 2.5 years after operation the patient has no problems, CT scan is without signs of recurrent diseases. The analysis of GIST was performed in agreement with contemporary knowledge. According to the references, our case report GIST of pancreas is the first of this kind (till 2003). PMID- 16315769 TI - Detection of new diagnostic markers in pathology by focus on growth-regulatory endogenous lectins. The case study of galectin-7 in squamous epithelia. AB - Lectins represent one of pivotal regulators of the cell proliferation The potential of galectin-7 as a new prognostic marker was studied in normal and transformed squamous epithelia of both ectodermal (epidermis, cornea vs. trichoepithelioma, basal and squamous cell carcinoma) and endodermal (vocal fold epithelium vs. carcinoma) origin. Studies on the cultured cells were also performed. Expression of galectin-7 seems to be connected to the process of stratification, no matter of origin of epithelium. Its expression is significantly reduced in malignant cells, thus galectin-7 might be a differentiation marker of epithelial malignancies. PMID- 16315770 TI - The effect of "mind on body". PMID- 16315771 TI - Ms. Reena Bose attends workshop on HIV/AIDS. PMID- 16315772 TI - Preparing nursing students for leadership positions. PMID- 16315773 TI - Self instructional module and cardiac angiography. Effectiveness of self instructional module on cardiac angiography for patients undergoing cardiac angiography in a selected hospital. PMID- 16315775 TI - Effectiveness of standard endotracheal suctioning technique on patients with mechanical ventilators. PMID- 16315776 TI - The privileges of being a 'behind the screen nurse'. PMID- 16315777 TI - A missing link in sarcoidosis. PMID- 16315778 TI - A critical assessment of treatment options for idiopathic pulmonary fibrosis. AB - BACKGROUND: To date, no management approach has proven to be efficacious for the treatment of idiopathic pulmonary fibrosis (IPF). Consequently, therapeutic options remain controversial and confusing for many clinicians. We sought to formally review available evidence on treatment options for IPF and to have a diverse panel of physicians rate the "appropriateness," "inappropriateness," or "uncertainty" of some of the available therapeutic options. METHODS: The RAND/UCLA Appropriateness Method was used to review and rate multiple clinical scenarios for the treatment of IPF. The panel was composed of nine physicians from geographically diverse areas who received a systematic review on the risks and benefits of commonly used treatments for IPF as background. RESULTS: A total of 324 clinical scenarios were rated: 25% as appropriate; 39%, uncertain; and 36%, inappropriate. The panel disagreed about 12% of the therapy indications in the final ratings, falling from 26% in the first-round ratings. CONCLUSIONS: Key themes emerged from the consensus process. Lacking evidence for a definitive therapy, it was considered most appropriate to enroll eligible patients in clinical trials and refer eligible patients for transplant evaluation. For patients without access to clinical trials, the committee was not unanimous regarding treatment recommendations. It was considered inappropriate for patients with a confident diagnosis of IPF to be treated with corticosteroids as the sole agent: corticosteroids should be used in conjunction with azathioprine. With progressive disease despite such combination use, there was agreement for the use of interferon gamma-1b in patients unwilling or unable to participate in available clinical trials. PMID- 16315779 TI - Role of the berylliosis-associated HLA-DPGlu69 supratypic variant in determining the response to beryllium in a blood T-cells beryllium-stimulated proliferation test. AB - BACKGROUND AND AIM OF THE WORK: Exposure to beryllium (Be) compounds may cause sensitization as revealed by blood T-cell proliferation against Be in a standardized Be-stimulated T-cell proliferation test (BeLPT). Further, susceptibility to Be hypersensitivity has been associated with the expression of HLA-DP allelic variants carrying a glutamate residue at position 69 of the beta chain (HLA-DPGlu69) in more than 80% of affected subjects and, at lower frequency, with other HLA-DP, -DQ and -DR alleles/polymorphisms. The aim of this study was to assess whether heterozygous or homozygous carriage of the HLA DPGlu69 marker may dictate the intensity of the T-cell response to Be in vitro. METHODS: The results of the blood Be-LPT, performed in a single laboratory on 165 subjects (86 Be-exposed controls, 38 Be-sensitized without lung granulomas and 41 berylliosis cases) identified at a single large Be production factory, were analyzed for their realtionship with the HLA-DPGlu69 status as determined by high resolution HLA-DP typing. RESULTS: Be-sensitized subjects carrying HLA-DPGlu69 presented a significantly higher T-cell response to Be (mean SI: 24.6 +/- 38.7) than the HLA-DPGlu69-negative subjects (mean SI: 11.8 +/- 6.6, p = 0.021). Furthermore, HLA-DPGlu69-positive subjects presented a higher frequency of positive Be-LPT tests (mean frequency 0.36 +/- 0.23) compared to the HLA-DPGlu69 negatives (mean frequency: 0.22 +/- 0.15; p = 0.002). HLA-DPGlu69 homozygosity was not associated with an increased response to Be in the BeLPT CONCLUSIONS: These results indicate that the expression of HLA-DPGlu69 determines higher T cell proliferation rates and more consistent resposes to Be in vitro in the BeLPT test, both in the homozygous and the eterozygous state, possibly leading to an underestimation of the numbers of HLA-DPGlu69-negative sensitized subjects within exposed populations. PMID- 16315780 TI - NOD2/CARD15 gene polymorphisms in idiopathic pulmonary fibrosis. AB - BACKGROUND: Micro-organisms, behaving in a non-infectious fashion, may be among the exogenous factor(s) believed to trigger idiopathic pulmonary fibrosis (IPF). One possible strategy to identify an individual's susceptibility to such microbial triggers, which are likely to be ubiquitous, is to investigate the molecular processes involved in their recognition. NOD2/CARD15 is a specific pattern recognition receptor protein, whose genetic variants have been previously associated with susceptibility to Crohn's disease. AIM: The aim of this work was to determine the frequencies of the three major NOD2/CARD 15 gene mutations (R702W, G908R and 1007fsinC) in a series of 76 subjects affected by IPF, and to compare them with those found in three groups of controls: a group with sarcoidosis (a disorder in which an involvement of the NOD2/CARD15 gene has already been investigated and rejected in different ethnic groups; 67 subjects) and two groups of healthy subjects (218 and 208 subjects, respectively), matched for gender, age, and ethnicity. RESULTS: We found no differences in frequencies of NOD2/CARD15 gene polymorphisms among the four groups investigated. CONCLUSION: We conclude that the NOD2/CARD15 gene is not likely to be involved in susceptibility to IPF in Italians. PMID- 16315781 TI - Neurotrophin system activation in bronchoalveolar lavage fluid immune cells in pulmonary sarcoidosis. AB - BACKGROUND AND AIM OF THE STUDY: Pulmonary sarcoidosis is a chronic granulomatous disease characterized by macrophage and CD4+ T-cell accumulation at the site of inflammation. Analysis of the cytokine network has substantially improved knowledge on immunopathogenesis of sarcoidosis. We hypothesize that neurotrophins (NTs), nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and NT 3, besides their importance in immune system activities, participate in chronic inflammatory disorders and in repair processes. METHODS: The expression of NTs and NT receptors was assessed in broncho alveolar lavage (BAL) macrophages, CD4 and CD8 T-cells, from 10 patients with pulmonary sarcoidosis, using molecular biology, Western blotting and immunocytochemistry. RESULTS: Increased levels of NTs and of high affinity NT receptor (Trks) transcripts and proteins in BAL macrophages, CD4+ and CD8+ T-cells from pulmonary sarcoidosis patients were demonstrated in comparison with healthy controls. Contrarily to healthy controls, in pulmonary sarcoidosis the expression of NGF was increased in alveolar macrophages as well as NGF and BDNF in CD4+ and CD8+ T-cells. An increased expression of TrkA, TrkB and TrkC receptors was also noticeable. Furthermore, BDNF expression in alveolar macrophages and NT-3 expression in the three different BAL immune cell populations investigated were induced during sarcoidosis. A significant correlation was observed between CD4:CD8 ratio, lymphocytosis, radiological stage and CD4 and CD8 NT expression. CONCLUSIONS: These findings suggest that NTs are exaggeratedly expressed in BAL immune cells in pulmonary sarcoidosis and may participate in the progression of disease modulating immune cell functions. PMID- 16315782 TI - Angiotensin-converting enzyme 2 (ACE2) haplotypes are associated with pulmonary disease phenotypes in sarcoidosis patients. AB - BACKGROUND AND AIM OF THE STUDY: Angiotensin II (Ang II) formation by angiotensin converting enzyme (ACE) or other enzymes has shown to exhibit profibrotic properties in a variety of fibrotic diseases. A homologue of ACE called ACE2 has been shown to counteract the formation of Ang II. Genetic variation in the components involved in Ang II formation may underlie the progression of pulmonary sarcoidosis. METHOD: Seven ACE2 SNPs, located on the X-chromosome, were investigated using SSP-PCR and haplotypes were constructed. Gender-matched analyses of sarcoidosis patients (80 males/64 females) and controls (110 males/218 females) were performed to correlate disease susceptibility and pulmonary disease phenotypes with ACE2 genotypes and haplotypes. RESULTS: ACE2 SNPs or haplotypes were not associated with susceptibility for sarcoidosis. Haplotype 4 was only present in sarcoid males without parenchymal involvement (frequency: 0.19) and absent in males with parenchymal involvement (p = 0.006; pcorr. = 0.05; degrees of freedom (df) = 1; OR = 0). No significant difference was observed between haplotype 4 frequencies in females with (0.08) or without (0.13) parenchymal involvement (p = 0.5). Although not significant after correction, analysis of the patient group with fibrosis showed that males with haplotype 5 (0.27) were predominant over those with haplotype 5 of the groups without fibrosis (0.03); p = 0.01; pc = 0.08; df = 1; OR = 11.4. Females with fibrosis vs. no fibrosis revealed no difference between haplotype 5 frequencies: 0.05 vs. 0.03; p = 0.37; pc = 1; df = 1. CONCLUSION: These results suggest that ACE2 might be involved in the progression of pulmonary sarcoidosis which may depend on gender. Subsequent studies using larger groups are needed to confirm these findings. PMID- 16315783 TI - Investigation of IL-18 and IL-12 in induced sputum of patients with IPF before and after treatment with interferon gamma-1b. AB - BACKGROUND AND AIM: Interleukin-18 (IL-18) has been identified as an interferon gamma (IFN-gamma) mediator, promoting a T helper 1 (Th1) response. Th1 response is characterized by increased expression of IFN-gamma, interleukin-12 (IL-12) and interleukin-18 (IL-18). The present study aims to evaluate the role of Th1 cytokines by monitoring changes in Induced Sputum (IS) samples, before and after treatment with IFN-gamma-1b in patients with IPF. PATIENTS AND METHODS: Fifteen patients with histologically confirmed IPF/UIP (12 male, 3 female) of median age 65 yr were prospectively studied. Ten healthy subjects (5 female, 5 male) of median age 61 yr served as control group. Patients were assigned to receive IFN gamma-1b 200 microg (15 patients) subcutaneously three times per week for 12 months. Induced sputum (IS) IL-12 and IL-18 levels were measured before and after IFN-gamma-1b treatment in IPF patients as well as in healthy controls, using ELISA immunoassay. RESULTS: The IL-18 levels were significantly higher in IPF samples before treatment than in healthy controls (57.05 +/- 6.9 pg/ml vs. 41.07 +/- 8.16 pg/ml, p < 0.05). A statistically significant decrease was detected in the IL-18 levels after IFN-gamma-1b treatment (57.05 +/- 6.9 vs. 42.8 +/- 5.1 pg/ml, p = 0.04). The IL-12 supernatant levels measured before and after IFN gamma-1b treatment were not significantly different. CONCLUSIONS: Our results may illustrate the potential role of IL-18 as an inflammatory molecule in the pathogenesis of IPF. Moreover, decrease of IL-18 levels in IPF patients, after 12 months of therapy could possibly be explained as IL-18 downregulation after IFN gamma-1b treatment. Extended studies are needed to determine the precise role of IL-12 and IL-18 during IPF. PMID- 16315784 TI - National survey on status of steroid therapy for cardiac sarcoidosis in Japan. AB - AIM: This investigation was undertaken to clarify the current status of steroid therapy for cardiac sarcoidosis in Japan. METHODS: A questionnaire survey was conducted throughout Japan concerning cases in which steroid therapy had been administered. Replies describing 52 cases (15 men, 37 women; mean age +/- SD, 59.8 +/- 14.5 years) were analyzed. RESULTS: Of the 49 patients whose New York Heart Association (NYHA) functional classification was reported, 29 (55.8%) were in class I; 13 (25.0%) class II; 4 (7.7%) class III and 3 (5.8%) class IV. The most common initial steroid dose (used in 35 cases, or 67.3%) was 30 mg/day or 60 mg on alternate days. In most cases (85.4%), this dose was continued for 1 month followed by tapering by 5 mg every 2 to 4 weeks until reaching the maintenance dose of 5 to 10 mg/day. Steroid therapy was reported to result in improvement in 54%, no change in 40%, and deterioration in 6%. CONCLUSION: This nationwide questionnaire survey indicated fairly uniform patterns of steroid therapy for cardiac sarcoidosis in Japan, with clinical improvement in over one-half of cases and possible stabilization in most others. PMID- 16315785 TI - Effect of beclomethasone dipropionate (BDP) as extrafine aerosol on bronchoalveolar lavage (BAL) lymphocytes in chronic sarcoidosis. AB - BACKGROUND AND AIM OF THE STUDY: Patients with pulmonary sarcoidosis can benefit from inhaled corticosteroids. In this study we assessed the effect of beclomethasone dipropionate (BDP), administered as extrafine, HFA(hydrofluoroalkane)-driven aerosol with high peripheral deposition, on bronchoalveolar lavage (BAL) lymphocyte numbers, as a marker of the disease. METHODS: Fifteen patients with newly-diagnosed pulmonary sarcoidosis (Stages I III) received either BDP 800 microg daily (n = 6) or placebo (n = 9) for 6 months in a parallel-group design. Before and after treatment, clinical and radiological states were assessed, bronchoalveolar lavage (BAL) performed, and the cellular composition of BAL fluid as well as cytokine production by BAL lymphocytes determined. RESULTS: BDP caused a decrease in the percentage of BAL lymphocytes in the BDP (p < 0.05) but not the placebo group. HLA-DR expression on lymphocytes was diminished after BDP (p < 0.05), while intracellular cytokine production by lymphocytes was not altered. Chest radiography suggested an improvement in the BDP group. There was also a rise (p < 0.05) in the diffusing capacity for carbon monoxide. CONCLUSIONS: Though based on a small group of patients, the present findings suggest that in patients with pulmonary sarcoidosis of stage II and minor functional impairment, inhalation of high doses of BDP as extrafine, peripherally deposited aerosol is associated with a reduction in the number of BAL lymphocytes, in parallel with improvements in other markers of the disease. PMID- 16315786 TI - Lung transplantation for end-stage pulmonary sarcoidosis: outcome in a series of seven consecutive patients. AB - BACKGROUND: From 1992 through October 2004, in total 379 lung transplantations (LTx) have been performed at this center. The aim was to describe the course of LTx in 7 consecutive patients with end-stage pulmonary sarcoidosis. METHODS: There were 5 men and 2 women. Figures are median (range). Pretransplantation data: Age at debut of sarcoidosis: 32 y (17-44). Duration of sarcoidosis until LTx: 13 y (10-30). None had significant extrapulmonary sarcoidosis. Chest X-ray showed stage III-IV sarcoidosis. All patients were NYHA class III-IV. Pulmonary function in per cent of predicted: FEV1 20% (15-75), FVC 43% (27-75), DLCO 26% (17-60). RESULTS: All patients had single lung transplantation (SLTx) at a median age of 51 y (45-55). The perioperative course was uncomplicated. Follow-up was 30 months (11-135). A 47-year-old woman died 22 months post-LTx due to bronchiolitis obliterans syndrome (BOS) with graft failure. Maximum pulmonary function tests at 2-6 months post-LTx: FEV1 57% (52-91), FVC 69% (63-110). Acute rejection episodes (A2-A3) were median 3 (0-4). BOS at follow-up: 1 patient has BOS grade 0-p, 1 patient grade 1, 2 patients had grade 2, and 2 patients had grade 3. Recurrence of sarcoidosis in the transplanted lung occurred in 3 patients 1-6 months after SLTx, without deterioration in lung function. CONCLUSIONS: Lung transplantation in patients with end-stage pulmonary sarcoidosis does not differ significantly with respect to morbidity or mortality compared with patients having other lung diseases. Recurrence of sarcoidosis in the transplanted lung occurs in approximately 50% of the patients, but apparently has no significant impact on lung function or on short-term outcome. PMID- 16315787 TI - Pulmonary involvement in an adult female affected by type B Niemann Pick disease. AB - We describe the case of a 33-year-old woman who presented with a pattern of diffuse micronodular opacities with centrolobular distribution at high resolution chest tomography (HRCT) performed after exposure to the smoke of a home fire. An abdominal CT scan showed the presence of 3 rounded hypodense lesions in the spleen parenchyma. A bronchoalveolar lavage (BAL) was performed, showing the presence of lipid laden cells in the aspirated fluid. A video-assisted thoracoscopic biopsy confirmed the presence of the foamy cells filling the alveolar spaces and showed a preserved background pulmonary architecture; the bronchiolar epithelium presented a cytoplasm with a particularly clear colour. Foam cells were present in the bone marrow aspirate too, that contained also sea blue histiocytes. These data induced us to consider the diagnosis of a metabolic storage disorder, in the first instance Niemann-Pick type B, that was confirmed by low acid lysosomal sphingomyelinase activity on cultured fibroblasts: 5.22 nmol/mg prot/h (n.v. 92 +/- 18.2). Lung involvement is an important complication of Niemann-Pick disease, presenting with the clinical and radiological features of a diffuse lung disease. PMID- 16315788 TI - Combination of 18F-fluoro-2-deoxyglucose positron emission tomography and magnetic resonance imaging in assessing cardiac sarcoidosis. PMID- 16315789 TI - The effects of fatigue and treatment with methylphenidate on sustained attention in sarcoidosis. PMID- 16315790 TI - Parotitis as first presentation of localized (initial phase) and generalized Wegener's granulomatosis. PMID- 16315791 TI - Treatment of pulmonary hypertension secondary to alveolar microlithiasis with Bosentan: a case report. PMID- 16315792 TI - Nurses split over physician assistants. PMID- 16315794 TI - Full prescribing raises nursing to a new level. PMID- 16315793 TI - Speak out now if you want to save the NHS. PMID- 16315795 TI - All change in primary care. PMID- 16315796 TI - Preventing cervical cancer by vaccinating against HPV. AB - There was widespread media interest in a report on the successful results from a trial of a vaccine against virus strains responsible for cervical cancer. This article examines the background to the report, considers the actual outcomes and comments on the significance and likely impact of the vaccine. PMID- 16315797 TI - Alzheimer's disease. PMID- 16315798 TI - Drug calculations. PMID- 16315799 TI - Understanding and dealing with parental vaccine concerns. AB - Parents often worry more about vaccines than the diseases they prevent. This article highlights the issues that commonly concern parents and suggests appropriate information that nurses can give to reassure them and promote vaccination. PMID- 16315800 TI - ACE inhibitors. PMID- 16315801 TI - Using light to treat age-related macular degeneration. AB - Age-related macular degeneration (AMD) is a potentially blinding disease that can be treated by verteporfin photodynamic therapy. This article discusses the specialist ophthalmic nurse's role in the assessment, treatment and support of patients with AMD, highlighting key elements in the care pathway and the benefit of prompt triage in safeguarding remaining vision. PMID- 16315803 TI - Education should not be a luxury. PMID- 16315802 TI - Costs and benefits of private finance initiative schemes. AB - The private finance initiative (PFI) is the biggest building programme in the history of the NHS. It aims to raise the quality of health care facilities by utilising the skills and expertise of companies in the private sector. This article outlines what PFI involves, how it works and the benefits to the NHS in raising the quality of health care facilities. PMID- 16315804 TI - New guideline on preventing and managing pressure ulcers. AB - New guidance on the prevention and management of pressure ulcers has been developed collaboratively by the National Institute for Health and Clinical Excellence (NICE) and the Royal College of Nursing. Julie Stevens and Will Gray provide the background to its development and discuss the key recommendations. PMID- 16315805 TI - Silver dressings: absorption and antibacterial efficacy. AB - Alan Lansdown and colleagues report on a small study that aimed to illustrate patterns of silver release and distribution in wound sites using non-invasive methods. It demonstrated that silver released into a wound bed is absorbed by exudate or dead tissue and that patients' health is not compromised PMID- 16315806 TI - Using finger-toe pulse oximetry to assess arterial blood flow. AB - National guidelines (RCN, 1998; Clinical Resource Efficiency Support Team, 1998; SIGN, 1998) suggest that an assessment of ankle brachial pressure index using Doppler ultrasound is necessary to assess arterial blood flow in patients with leg ulcers or healed ulcers. Robin Cooper describes a newer, more simple test of arterial blood flow using a pulse oximeter, and discusses the advantages and disadvantages of each method. PMID- 16315807 TI - Understanding wound dressings: hydrocolloids. AB - In the third article in this series looking at wound dressings, Jacqui Fletcher describes hydrocolloid dressings. PMID- 16315808 TI - Nursing a patient with frostbite. AB - Frostbite is a condition that is associated with people living in countries with an extremely cold climate. However, it can affect people in the UK who are involved in winter sports, and is also associated with the homeless, people with a psychiatric illness, and those who misuse drugs and alcohol. Angela Davies describes the assessment and management of a patient with frostbite and identifies the challenges of managing these complex tissue injuries PMID- 16315809 TI - Boosting the health of the workforce. PMID- 16315810 TI - Research for best nursing. PMID- 16315811 TI - Needle phobia--changing venepuncture practice in ambulatory care. AB - Needle phobia is a term used in practice to describe an anticipatory fear of needle insertion. A proportion of children display high levels of fear, pain and behavioural distress when exposed to, or anticipating, needle insertion. A difficult routine venepuncture in our ambulatory care unit led staff to review practice and develop a three-step approach to overcoming 'needle phobia': relaxation, control and graded exposure. These developments have resulted in the unit becoming a local referral centre for children and young people between the ages of 5-19 years with this problem. Time and skill are needed to prevent or overcome this distressing problem which can be caused by health care professionals not listening to children and young people. PMID- 16315812 TI - Hospice care in Romania. PMID- 16315813 TI - Developing numeracy in child branch students. AB - The Nursing and Midwifery Council (2002a) reports that one of the most common examples of professional misconduct relates to the failure to administer medicines safely. In educating child-branch nurses, we have a duty to enhance students' calculation skills so that they are safe and can provide the best possible care to children. This paper presents a framework for numeracy within a child-branch curriculum, describing a practical approach to developing students' skills in the mathematical calculations they are likely to encounter in clinical practice. A pass mark of 70 per cent in relevant numeracy tests is considered too low for child branch students. Early identification of mathematical ability and provision of relevant tutorial support will ensure that the majority of students achieve the required standard. PMID- 16315814 TI - Using books to prepare children for surgery. PMID- 16315815 TI - Parental satisfaction with a nurse-led emergency assessment unit. AB - Parents of children dischared from the paediatric emergency assessment unit of a large regional children's unit were surveyed to discover their perceptions of the care experience and post discharge support. Data from 40 parents revealed that the family journey through the unit was generally positive, although not without problems. The majority of children were referred from their general practitioner, and most were seen by a paediatrician within an hour of arrival. Morning-after phone calls and 24-hour access after discharge increased satisfaction. However, nearly a third of families reported they were not shown the facilities of the unit and just under half mentioned lack of privacy as a concern. An action plan to address these issues has been subsequently implemented and will be evaluated periodically. PMID- 16315816 TI - Restraining children for clinical procedures. PMID- 16315817 TI - Obtaining urine for culture from non-potty-trained children. AB - The accurate diagnosis of urinary tract infection (UTI) in young children is important for initial management and the prevention of long-term damage to the developing kidney. Various methods are used for urine collection from the non potty-trained age group, including catheterisation, suprapubic aspiration, clean catch sampling, adhesive bag sampling and urine collection pads. Research literature was examined to identify which method is most effective. The themes that evolved from the review were parents' and nurses' views on ease of use, reliability of sample obtained and cost-effectiveness. Although not the most convenient method, clean-catch gives more reliable results making it also the most cost effective approach. Change management strategies are required to overcome barriers to research implementation and to ensure best practice. PMID- 16315818 TI - Reliability and validity in practice: assessment tools. PMID- 16315819 TI - The "animal rights" movement's cruelty to humans. PMID- 16315820 TI - The Walter C. Randall Lecture and an agenda for ethics. PMID- 16315821 TI - Looking beyond the surface: a beginner's guide to the surgical literature. PMID- 16315822 TI - Protected carotid artery stenting (PCAS): a short medical technology assessment. AB - After a period of experimenting with angioplasty and stenting, carotid artery stenting under embolic protection (PCAS) is becoming a viable alternative for carotid endarterectomy (CEA). A standard literature review showed that, at January 2005, there was no evidence that PCAS is more effective than CEA. The high costs of stent and protection device makes PCAS then inferior to CEA. PCAS may be the sole possible option in patients with symptomatic carotid artery stenosis unfit for surgery, where the high risk of stroke overrides uncertainty about health effects and overrides cost-effectiveness. These are a few patients per year in Belgium. Several randomised controlled trials comparing PCAS and CEA are now recruiting patients. To have answers on key questions of cost effectiveness, it is of paramount importance that these trials recruit and publish rapidly. The KCE (Belgian HealthCare Knowledge Center/Centre Federal d'Expertise des Soins de Sante/Federaal Kenniscentrum voor de Gezondheidszorg) therefore advises cooperation with these trials. Outside these trials and compassionate use in the few symptomatic patients unfit for CEA, the use of PCAS raises serious ethical questions. PMID- 16315823 TI - Accreditation and quality approach in operating theatre departments: the French approach. AB - Since 1996, French health establishments are subjected to a process of evaluating the quality of care, called "accreditation". This process was controlled by ANAES, which, after January 1st, 2005 became the Haute Autorite de Sante (HAS). The accreditation is characterized by a dual process of self-assessment and external audit, leading to four levels of accreditation. In spite of requiring a time-consuming methodology, this approach provides an important means of consolidating the development of the quality approach and re-stimulating the compliance of establishments with standards of safety and vigilance. The professional teams of many French operating theatre departments have been able to use the regulatory and restricting framework of accreditation to organize quality approaches specific to the operative system, supported by the organizational structures of the department such as the operating suite committee, departmental boards and the steering group. Based on quality guidelines including a commitment from the manager and operating suite committee, as well as a quality flow chart and a quality system, these teams describe the main procedures for running the operating theatre. They also organize the follow-up of incidents and undesirable events, along with the risks and points to watch. Audits of the operative system are planned on a regular basis. The second version of the accreditation process considerably reinforces the assessment of professional practices by evaluating the relevance, the risks and the methods of managing care for pathologies. It will make it possible to implement assessments of the health care provided by operating theatre departments and will reinforce the importance of search for quality. PMID- 16315824 TI - The law on e-Health: draft proposal for an electronic medical prescription. PMID- 16315825 TI - The challenge of electronic medical prescriptions to the rule of confidentiality and to the respect of patient's privacy. PMID- 16315826 TI - The protection of the patient's private life: a vast normative landscape. First part. AB - The debate on the protection of people's private life is stronger than ever, particularly in the health care sector, where choices have to be made in order to decide between the conflicting interests at stake. This Article gives an overview of the general provisions that aim, for the time being, at guaranteeing this protection to the patient. His or her right to respect for private life, which aims at protecting not only the confidentiality of his or her private realm and relationships with others, but also his or her freedom of choice regarding this private realm and these relationships, is indeed expressed, in various forms, in a large number of texts of every possible origin. It would be a mistake to regard these as a heap of redundant rules. These texts can be distinguished according to their content, the time of their elaboration and their authors. Although complex, the normative structure that they form is not void of effectiveness. A next article will examine how they are extended and supplemented by provisions trying to answer, at the level of the respect of private life, to the growing use of computers in medical practice. PMID- 16315827 TI - The protection of the patient's private life: the computer challenge. Second part. AB - Today, medical practice is invaded by a growing number of technologies of all kinds, among which computer techniques have an important place. Although they have significant advantages, for instance in terms of medical record management, they give rise to several problems, particularly concerning the confidentiality of the patient's data with regards to third party. A great number of specific provisions, complementary to the general texts protecting private life (examined in the first part of this two parts article), endeavour to solve these problems. It is true that these provisions are recent, have various origins and often appear as rules difficult to understand. Yet, they are partially inspired by a common logic. Relying on these common features, the authors make two suggestions for the future, in order to avoid that the growing computerisation of medical practice eventually destabilises the health care relationship: a) Any dictatorship of confidentiality must be rejected. b) Stimulating a sense of professionalism is most likely the way to avoid an anarchic and unrealistic development of rules aimed at regulating the health care relationship. PMID- 16315828 TI - Pelvic anatomy for colorectal surgeons. AB - INTRODUCTION: recent advances in rectal surgery include total mesorectal excision and preservation of the autonomic pelvic nerves, so that colorectal surgeons have to get some information on the embryology of the rectum, on the complex anatomy of the pelvic floor and on the distribution of lymphatic nodes. Embryology includes the formation of the hindgut during the first month of the embryo and the subsequent formation of the rectum and anal canal. The mesorectum contains the blood vessels and the lymphatic nodes. To totally excise the mesorectum, the surgeon should follow the "holy plane" described by Heald, between the perirectal fascia and the pelvic fascia. Doing this, the surgeon has the best chance to preserve the autonomic pelvic nerves that comprise the superior hypogastric plexus, the right and left hypogastric nerves and the right and left inferior hypogastric plexuses. PMID- 16315829 TI - The usual causes of left ventricular outflow tract obstruction below the aortic valve in normal ventriculoarterial connection: review of the physiopathology and surgical implications. AB - Subaortic narrowing leading to left ventricular outflow tract obstruction (LVOTO) is a significant pathology that may be encountered pre- or postoperatively in both acquired or congenital cardiac disease. Through a review of the morphological features and mechanisms in the different clinical situations, the anatomic, dynamic and mixed forms of subaortic obstruction are emphasized. Knowledge of the substrate of LVOTO, its physiopathology and natural history allows to plan accordingly the surgical repair and the postoperative management. PMID- 16315830 TI - Infected aortic aneurysms. PMID- 16315831 TI - Outlook of non operated type B aortic dissection with special reference to the incidence of degenerative abdominal aortic aneurysm (AAA). One center study. AB - OBJECTIVES: to determine the value of pharmacological treatment of type B aortic dissection (B AD) in face of new forms of treatment. DESIGN: this is a retrospective study of the period from 1990 to 2000. Files of 81 patients have been reviewed and completed by questionnaires. RESULTS: Two B AD died after admission without any treatment, 10 were operated on with 7 discharged alive (group I); 69 received hypotensive agents and beta-blockers, 65 were discharged alive (group II). Late mortality of the group I is 3/7, not related with B AD. Late mortality after mean follow-up of 56.8 months is 27/65 with 4/27 related to B AD (4 ruptures, 2 operated on). Non fatal secondary surgery amounts 5 in 4 patients. Total B AD aortic events comprise 8/65 patients. Type A AD were operated on successfully (8 : 4 before B AD, and 4 after B AD). Degenerative abdominal aortic aneurysms were present, operated (9) or not (3), in the history of patients and 3 more appear subsequently. At 10 years, actuarial survival is 40% +/- 18. CONCLUSION: in non-complicated cases of B AD, medical treatment is a reasonable choice, provided that a strict follow-up of the thoracic abdominal aorta is performed. PMID- 16315832 TI - Pedal bypass for limb salvage. AB - BACKGROUND: Pedal bypass grafting is often the only method of limb salvage in patients with chronic critical lower limb ischemia due to atherosclerotic obliteration of the crural arteries, including patients with diabetic foot gangrene. It involves arterial reconstruction with distal anastomosis to one of the pedal arteries. MATERIAL AND METHODS: Between January 2000 and June 2004, 54 pedal bypasses were performed in 53 patients with chronic critical lower limb ischemia. Forty-seven (87%) patients had gangrene or ischemic ulcer, 36 (68%) had diabetes. In some of the patients (16.7%), previous percutaneous transluminal angioplasty (PTA) of the crural arteries had failed. Preoperative angiographic findings were unsatisfactory in the majority of the patients; the plantar arch was not visualized in 36 (66.7%) limbs. RESULTS: In the period investigated (54 months) 11 grafts (20.4%) failed. Early thrombectomy resulting in long-term graft patency salvaged five limbs. One limb with graft occlusion occurring after foot ulcer healing was also salvaged. However, one amputation had to be performed despite a patent graft. The perioperative mortality rate was 3.8%. Cumulative primary and secondary graft patency rates and limb-salvage rates at 54 months were 76%, 78% and 81%, respectively. CONCLUSION: Pedal bypass grafting is a safe method with very good long-term outcomes. The absence of the pedal arteries or plantar arch on preoperative angiograms need not be taken as a contraindication to pedal vascular reconstruction. In discussions on the plantar arch it is recommended to discriminate between its actual absence and a mere "angiographic" absence. PMID- 16315833 TI - Closure of atrial septal defects: is there still a place for surgery? AB - BACKGROUND: The purpose of this study is to assess the current management of atrial septal defect closure in an era of increasing feasibility of transcatheter device occlusion. METHODS: Atrial septal defect (ASD) closure was performed surgically through complete sternotomy in 165 patients (group 1) and through partial inferior sternotomy in 53 patients (group 2). Transcatheter device occlusion was achieved in 82 patients with only ASD type II and patent foramen ovale (group 3). RESULTS: Overall complications were minor and more frequent in group 1: 26.7% versus 13.2% in group 2 and 14.6% in group 3 (p = 0.04). Compared to complete sternotomy, a partial sternotomy led to less chest tube loss (7.1 +/- 2.9 versus 11.6 +/- 14.5 ml/kg) (p < 0.05) and less postoperative pericardial effusion (11.3% versus 13.5%)(p = 0.55). ASD closure was effective in 99.4% in group 1, 100% in group 2 but only in 86.6% in group 3 (p < 0.05). Two major complications of device implantation required early surgery: 1 femoral arteriovenous fistula and 1 device embolization. Hospital stay was significantly shorter in group 3, as well as in group 2 compared to group 1 (8.3 +/- 4.2 versus 5.9 +/- 1.1 versus 2.1 +/- 7.3 days) (p < 0.05). Midterm results were excellent, with only 1 non-cardiac death and 1 re-operation for residual shunt in group 1, and 1 device removal for thrombosis in group 3. CONCLUSION: Transcatheter device occlusion has become an established treatment for ASD closure, achieving optimal results in older children and adults with anatomically suited ASD type II and PFO. However, a partial inferior sternotomy offers a valuable and complementary operative approach for all ASD variants, maintaining the predictable success of surgery, with the obvious advantages of minimal access in terms of morbidity, cosmetics and hospital stay. PMID- 16315834 TI - Thoracoscopic talc pleurodesis for the treatment of spontaneous pneumothorax. AB - BACKGROUND: During the Belgian Surgical Week in May 2004 some controversy existed on the optimal treatment of spontaneous pneumothorax. Doubts raised about the safety of talc in performing pleurodesis because of reported complications. METHODS: A retrospective analysis of a consecutive series of thoracoscopic pleurodesis with talc was performed. Patients operated for spontaneous pneumothorax were analysed focusing on complications and freedom of recurrence. A literature search was performed on complications from the use of talc to treat pleural disease. RESULTS: From September 1999 till August 2004 forty-one patients had a thoracoscopic pleurodesis with talc. In 21 patients this was performed for recurrent malignant pleural effusion and in 20 for spontaneous pneumothorax. In 5 of these 20 patients we faced a secondary spontaneous pneumothorax. In seven patients an apical pulmonary wedge resection of bullae was performed. No intra operative or serious postoperative complications were seen in these patients. All drains were removed after 4 to 6 days. No episodes of respiratory insufficiency occurred. No recurrence was encountered during a mean follow-up of 22.7 months. CONCLUSION: Thoracoscopic talc pleurodesis for spontaneous pneumothorax was effective and safe in our experience. In the literature no convincing evidence against the use of talc to treat pleural disease was found. Although some cases of ARDS are attributed to the pleural administration of talc, the incidence of complications after talc poudrage appears to be low. PMID- 16315835 TI - The influence of the implant size on the outcome of unconstrained total knee arthroplasty. AB - INTRODUCTION: The influence of implant size on the results in unconstrained total knee arthroplasty has never been investigated before. PATIENTS AND METHODS: 133 patients with unconstrained total knee arthroplasty were examined (HSS Score, Knee Society Score) with consideration of the different sizes of femoral and tibial components and the thicknesses of the tibial inlays. The medium follow up was 41.2 months (8 months to 74 months). RESULTS: Although the different component sizes gained different results, the differences were not significant. The combination of two component sizes did not lead to a significant change of the results. The results of the Knee Society Score seemed to become worse with thinner polyethylene tibial inlays although these differences were not significant. CONCLUSIONS: In conclusion, implant size had no significant influence on the outcome of total knee arthroplasty. The combination of two implant sizes showed no disadvantage. PMID- 16315836 TI - The role of pre-operative B mode ultrasound in the evaluation of the axillary lymph node metastases in the initial staging of breast carcinoma. AB - The lymph node status of a breast cancer is one of the main prognostic criterias. This status is very important to determine the therapeutic approach. Physical examination alone is not sufficient to assess axillary metastases. Mammographic examination can give us an idea about breast cancer and axillary involvement. Ultrasonographic evaluation can improve the sensitivity of clinical and mammographic examination in assessing axillary lymph node status. 42 patients operated on for breast cancer between January 2000-January 2003 were included in this prospective study. In the study, we used axillary B mode ultrasound to evaluate the axillary lymph nodes. There are several sonographic features to categorize them. Axillary B mode ultrasound was performed to evaluate the axillary lymph nodes for metastatic involvement. In the evaluation of lymph nodes, the sonographic criteria were centric echogenity, thickening of cortex, length/width ratio (L/W) and the diameter of lymph nodes. Hyperechogenic hilus was accepted as a benign finding. The thickening of the cortex less than 50% of the thickening of the centric echogenic hilus was also accepted as a benign finding. L/W ratio below 2 and parameters above 2 cm were accepted as malignant findings. 168 lymph nodes in 42 patients were evaluated pre-operatively with axillary B mode ultrasound. As a result, these lymph nodes were defined as benign in 19 patients (45.2%) and malignant in 23 patients (54.8%). Axillary lymph node status was found as benign in 18 patients (42.9%) and malignant in 24 patients (57.1%) pathologically . Comparative results of ultrasound and axillary lymph node status can be seen on Table III. As a result, the sensitivity of axillary B mode ultrasound to show the metastases was found as 79.1%, specificity was 77.7%, positive predictive value 82.6% and negative predictive value 73.6%. We think some better results may be obtained in the future and these developments may affect the surgeon's decisions concerning axillary dissection for breast cancer operations. PMID- 16315837 TI - Comparison of direct trocar and veress needle insertion in the performance of pneumoperitoneum in laparoscopic cholecystectomy. AB - Veress needle insertion, direct trocar insertion and open technique are different methods of establishing pneumoperitoneum to perform a successful laparoscopic procedure. We conducted this study to compare the use of a Veress needle and direct trocar insertion to create pneumoperitoneum. 274 laparoscopic cholecystectomy operations were evaluated. There were no significant differences in the age and gender between Veress needle and direct trocar entry groups. In this study we have seen that the complication rate while performing pneumoperitoneum by direct trocar entrance was less than in Veress needle usage. Direct trocar entrance also reduces the operation time. In laparoscopic cholecystectomy the direct trocar entrance method is a more reliable and less time consuming method than Veress needle usage. PMID- 16315838 TI - Noncurative total gastrectomy and oesophagogastrectomy in the treatment of advanced gastric carcinoma in a country with high incidence. AB - The role of extensive resectional surgery, including total gastrectomy for the palliation of advanced gastric cancer is controversial. This study shows operative results with complications and mortality occurring after total gastrectomy in patients with advanced stage gastric carcinoma. The study included 83 (48 males and 35 females, median age was 54.6 +/- 11.4 years) patients who underwent palliative total gastrectomy or oesophagogastrectomy (distal oesophagectomy in continuity with total gastrectomy). The reason for nonradical treatment was a too locally advanced disease. There was no case of carcinoma without serosal extension. Only five patients were free of histological lymph node metastases. A total of 72 (86.7%) early postoperative complications, including 17 self-limited wound complications, and 21 pulmonary complications were noted. Dehiscence of the oesophagojejunal anastomosis was noted in 7 patients, 3 of whom subsequently died. A total of 8 (9.6%) patients died in the postoperative period. The mean survival period was 12.8 +/- 0.8 months for all patients. It was 18.16 +/- 2.04 months in stage IIIA patients, 13.37 +/- 0.79 months in stage IIIB, and 7.51 +/- 0.97 months in stage IV patients. Total gastrectomy is a relatively safe procedure even when performing as a palliative procedure, with acceptable mortality and low lethal complication rate, and should be considered an alternative option in palliative treatment of advanced gastric cancer. PMID- 16315839 TI - Fibromuscular dysplasia of the superior mesenteric artery--case report and review of the literature. AB - Fibromuscular dysplasia is a multifactorial arteriopathy most commonly affecting the renal and carotid arteries. In this report we present a case of visceral artery involvement, causing occlusion of the superior mesenteric artery and celiac trunk and resulting in visceral ischemia. Treatment consisted of superior mesenteric artery reimplantation. Visceral artery FMD can present as occlusive or aneurysmal disease and treatment depends on patient characteristics and symptoms. PMID- 16315840 TI - Multinodular goitre with thyroid hemiagenesis: a case report and review of the literature. AB - Thyroid hemiagenesis is a very rare abnormality, in which one thyroid lobe fails to develop. Most of the patients have an associated thyroid disease. The true prevalance of thyroid hemiagenesis is not known, but it is estimated to be 0.02% in normal children. We report a forty-four-year-old female patient with a multinoduler goitre in the right lobe, associated with hemiagenesis of the left lobe. PMID- 16315841 TI - Simultaneous bilateral breast cancer in a male: a case report and review of the literature. AB - A 63-year-old male presented with a bilateral breast carcinoma. Both axillae showed metastases. The patient was treated with a bilateral modified breast amputation. Study of the literature shows that synchronous bilateral breast cancer in males is uncommon. Even in a male with changing breast tissue a carcinoma has to be considered. In male breast cancer, adherence to diagnostic and treatment guidelines for female breast cancer is generally advocated. In males, hormonal causes have to be excluded. PMID- 16315842 TI - Resection of extrahepatic hepatocellular carcinoma metastasis can result in long term survival. AB - Hepatocellular carcinoma (HCC) is one of the most common primary cancers in the world and the third most common cause of cancer mortality world-wide. Surgery is the gold standard in the treatment of patients with HCC. The prognosis is mainly determined by the underlying liver disease and recurrent rates. In the Western World, up to 30% of the patients with HCC have a non-cirrhotic liver. The main prognostic factor in this special group of patients are the recurrences. Most recurrences are intrahepatic; however, 30% of the recurrences are extrahepatic. The role of resection in case of intrahepatic recurrences is widely accepted, particularly in the non-cirrhotic liver. The role of resection in extrahepatic HCC recurrences is not well established and unknown among many physicians. We present two patients with HCC in a non-cirrhotic liver with extrahepatic recurrences and long-term survival after resection. The corresponding literature support an aggressive approach in case of extrahepatic HCC recurrence in selected cases: resectable metastasis, preserved liver function, absence of intracranial metastasis and control of the primary tumour. Further research is warranted because of the limited number of reports and the absence of randomized trials. PMID- 16315843 TI - Paravertebral schwannoma with high uptake of fluorodeoxyglucose on positron emission tomography. AB - A paravertebral mass was discovered in a 27-year-old woman, while investigating a painful shoulder and arm. CT, MRI and fine needle aspiration cytology (FNAC) pointed in the direction of a benign mass, but positron emission tomography (PET) showed a high uptake of [(18)F]fluorodeoxyglucose (FDG), which was indicative of a malignant lesion. Pathological analysis of the thoracoscopically resected tumour gave us the final diagnosis of a benign schwannoma. This report demonstrates that a high uptake of FDG in a non-malignant mediastinal tumour is possible. PMID- 16315844 TI - A completely displaced Salter-Harris I fracture of the distal radial epiphysis with intact ulna: a case report and review of the literature. AB - We present a 12-year-old boy with a displaced fracture of the distal radial epiphysis type I according to the Salter-Harris classification. The distal radial epiphysis was completely dorsally displaced, while the distal ulna remained intact. The injury was result of a fall on the patient's outstretched hand. Closed reduction was applied and the forearm was immobilized with a long arm cast. In the radiological re-examination a week later, a complete re-displacement of the fracture was detected. Open reduction followed and interposed flexor tendons (flexor pollicis longus and flexor carpi radialis) were found in the anatomical position of the distal radial epiphysis. The displaced epiphysis was reduced and fixed with two Kirschner wires and a long arm cast for a period of 6 weeks. The follow-up examination 2 years later showed that the movement range of the wrist joint was not limited and skeletal growth of the radius was not disturbed. The failure of closed reduction in these fractures is due to anatomical obstacles such as periosteum, flexor tendons and pronator quadratus interposition. Repeated forceful manipulations to achieve closed reduction must be avoided because of the potential for a number of complications, such as growth arrest, compartment syndrome, and avascular necrosis of the epiphysis. PMID- 16315845 TI - Fixation of a periprosthetic humeral fracture with CCG-cable system. AB - We report a case of periprosthetic fracture of the shoulder, treated operatively. A special plate/cable system (ECG) was used with an excellent outcome. This easy technique is recommended for such fractures. PMID- 16315846 TI - Destroyed lung due to aspiration of phenolphthalein. AB - A 76-year-old female, presented with a recurrent cough and haemoptysis after an aspiration of a 'grain de Valls'. Chest radiography and computed tomography demonstrated an atelectasis of the medial segment of the right middle pulmonary lobe. Bronchoscopy was negative for any malignancy but the onco-PET-scan showed a pathologic hypermetabolic lesion in the right middle lobe. A lobectomy was performed and pathology revealed a 'destroyed lung' with images of foreign body reactions and microabscesses without any sign of malignancy. Postoperatively, clinical improvement with complete resolution of the symptoms was observed. With this case report we demonstrate the irritative and clastogenic characteristics of phenolphtalein, the main active ingredient of the 'grain de Valls'. This case also illustrates an example of a false positive PET-scan. PMID- 16315847 TI - Tunga penetrans. A case report and review of the literature. AB - Tungiasis is an infection caused by infestation of Tunga Penetrans into the epidermis. Although, at the moment, presentation of this condition seems rare in Europe and Northern America, tungiasis is highly endemic in other parts of the world. We describe a case of Tunga Penetrans infection in the foot of a 22-year old Dutch female returning from Kenya, and present a review of the literature. Now that intercontinental travel is increasing, physicians in other parts of the world will be expected to recognize and treat this exotic infection. Knowledge of human tungiasis is mandatory in order to recognize the symptoms, achieve correct diagnosis and ensure adequate medical treatment to our patients. PMID- 16315848 TI - Atypical cause of forearm skin ulceration in a leukaemic child: mucormycosis. A case report. AB - Primary cutaneous mucormycosis is an uncommon, deep and aggressive fungal infection occurring mainly in immunosuppressed or diabetic patients. Rapid diagnosis and therapy are necessary to prevent a fatal outcome. An eight-year-old leukaemic child presented with a dark necrotic ulcer on the volar-ulnar aspect of the left forehand. The lesion had developed over seven days, beginning as a vasculo-haemorrhagic erythematous plaque. There was no known history of trauma to the area, but skin necrosis from external compression caused by an intravenous line could not be completely ruled out. The lesion rapidly progressed to a 5 x 8 cm painful necrotic ulcer with an erythematous border. Treatment with ichthyol dressing and intra-venous antibiotherapy failed to improve the condition. Wide debridement was performed and specimens were sent for microbiology and pathology examinations. Microscopic examination demonstrated broad, irregularly walled, non septate fungal hyphae that were consistent with Mucor. Amphotericin B was administrated intravenously (1 mg/kg/day) and hydrogel and hydrophile adhesive polyurethane foam dressings were applied. After 12 days, the soft tissue defect was covered with a split-thickness skin graft, harvested from the lateral aspect of the thigh. Diagnosis of this infection is based on complete histopathological and microbiological studies. Awareness, and a high index of suspicion are required because of the potential fulminant and fatal course. PMID- 16315849 TI - [Anatomy of the splint bones and their surrounding area particularly in consideration of fascial attachments]. AB - This study concerns the vestigial metacarpal and metatarsal bones, so called splint bones. The soft tissues, which stabilize the splint bones were carefully examined. The palmar metacarpal/plantar metatarsal fascia, the metacarpal/metatarsal interosseous ligaments and the fibres, which exit distally from the splint bone, were examined in depth. Cadaveric distal limbs were collected from 5 slaughtered or euthanized horses. The legs were examined both, microscopically and macroscopically. The palmar metacarpal/plantar metatarsal fascia is a very stable structure, which connects the splint bone with its environment. The connection between splint and McIII/MtIII, established through the metacarpal/ metatarsal interosseous ligaments, showed impressive differences between specimens examined. Macroscopic examination of the soft tissue in the distal region showed a band structure, which originates from the distal end of the splint bone and extends in dorsal and distal direction towards the proximal sesamoid bone. PMID- 16315850 TI - [Investigation about the clinical use of brushite- and hydroxylapatite-cement in sheep]. AB - For future clinical use as synthetic bone replacement, an injectable brushite (chronOS-Inject) and hydroxylapatite-(Biobon) cement were compared in a drill hole model in 10 sheep over time at 2, 4, 6, 8, 16 and 24 weeks. Results were compared regarding their practical use, biocompatibiliy, resorption mechanism and subsequent new bone formation. The cements were filled into drill holes (psi 8 x 13mm) of the proximal and distal humerus, and femur and the samples evaluated macroscopically, radiologically and microscopically including histomorphometrical quantification of percentages of new bone, fibrous tissue and remnants of cements. The cement area decreased continuously from 2 to 24 weeks with chronOS Inject, as well as the area of granules. Inversely, the subsequent new bone formation increased from 2-24 weeks accordingly. With Biobon the cement area decreased slower between 2 and 24 weeks, and the new bone formation was less. Both cements were well integrated into the bone in long bones. chronOS-Inject demonstrated good biocompatibility and was almost completely replaced through bone within 24 weeks. Biobon was resorbed considerably slower and initially a slight inflammatory reaction including bone resorption was observed within the adjacent host bone. PMID- 16315851 TI - [The significance of elbow dysplasia (ED) for breeding in Bernese Mountain Dogs in Germany]. AB - Results from the elbow dysplasia screening program in Bernese Mountain Dogs of Germany were analysed in respect to its relevance for genetic evaluation and breeding. In total 2677 gradings were used. The grading was performed radiographically according to the recommendations of the International Elbow Working Group (IEWG). 75.8% of the dogs were free from visible dysplastic signs, 10.8%, 6.8% and 5.5% were classified to be of Grade 1, 2 and 3, respectively. 1.1% were classified as a borderline case between ED-free and Grade 1. A slight reduction of ED could be observed over years. Males had a 3.1 higher rate of dysplasia than females. Treating ED as a numerical trait, coded proportional to the severity of clinical relevant signs, gave a heritability estimation of 0.188 and a maternal effect of 0.07. From different mating combinations it was found, that ED average was higher in the progeny if one mate was affected but there was no increase in the prevalence with increasing grade of ED in the affected mates. About 10% more affected dogs could be observed, if one mate is affected. Heritability for dichotomic coding the trait was found to be 0.20. From these results can be suggested, that the differentiation in various grades of ED to describe the clinical relevance for the specific dog is helpfull, however, from a breeders point of view, dogs with ED should be treated equally for genetic evaluation. PMID- 16315852 TI - Alveolar echinococcosis in the zoological garden Basle. AB - Alveolar echinococcosis (AE) is a rare, but potentially severe zoonotic disease caused by Echinococcus (E.) multilocularis. Recent findings indicated an increasing importance of AE for non-human primates living in regions endemic for E. multilocularis. The death of five cynomolgus monkeys (Macaca fascicularis) and a lowland gorilla (Gorilla g. gorilla) due to AE raised concern about the incidence of this parasite in the Basle Zoo. Consequently, a project was initiated to investigate the prevalence amongst an affected group of cynomolgus monkeys, as well as in foxes and mice. Three out of 46 monkeys were seropositive for E. multilocularis antigen. In two of these monkeys and in another three animals, which were seronegative, structures compatible with metacestodal cysts were observed using ultrasonography. Seven out of 35 free roaming foxes caught at the zoo were positive for an intestinal E. multilocularis copro-antigen ELISA, four of them shed taeniid eggs simultaneously. No lesions compatible with AE were present in 50 necropsied mice from the zoo area. These results indicate that the fox population is a potential source to introduce E. multilocularis and may thus represent a risk for the zoo animals. PMID- 16315853 TI - [Immigration of dentists and its effects on the dental profession in Hungary]. AB - There were a very remarkable ethnic and demographic shift in the social composition of dental health care providers in Hungary after the collapse of the old regime in 1989. Hungary is surrounded by states with large ethnic Hungarian minorities blocked in their free migration up to the late 1980's. Although the non-educated members of the Hungarian diaspora did not leave these areas in mass numbers, a great number of highly educated people (among them preeminently health care professionals) moved to the mother state mainly for economic reasons. Dentistry comparing to other medical professions has a special financial advantage because of the considerable share of private financing in the Hungarian dental business. Unfortunately, no statistical analysis was made on the impact of this professional migration in the last 15 years. The present article based on a cross-sectional survey conducted at the end of 2004 demonstrates the proportion of immigrants (mean 7.81 %) among the total number of dental health care providers (5670 to a 10.2 million population) There are 2764 area bounded solo practices contracted to the National Health Insurance (NHI), the other 2907 professionals are engaged in the free market business, and/or they are public employees of the universities with dental faculty as well. As expected, the share of immigrants is higher in NHI-practices, but it varies in a wide range (1.9 - 23.6%) amidst the different counties (19 units plus the capital). Nevertheless, the immigrants may also participate in the free market segment of the dentistry. The variations can be explained by geographic, internal migrational and local economic reasons. PMID- 16315854 TI - [Neurophysiological background of pain in the orofacial area: review of the literature]. AB - The article presents an overview on the peripheral and central neural mechanisms underlying pain in the orofacial area. First a definition of pain and a description of general aspects of orofacial pain are presented. Characteristics of acute and chronic pain are also described. The study highlights the sense organs, the molecular mechanisms and categories of primary afferents involved in peripheral events of orofacial pain. After describing the brain-stem nuclei participating in trigeminal pain and their functional role, primary afferents involved in nociceptive sensation from the tooth pulp, explanations of dentinal sensitivity and differences between the brain-stem endings of primary afferents among different species are discussed in details. The role of higher brain centres, with a special emphasis on the thalamus and somatosensory cortex in the development of orofacial pain sensation is considered. The last section provides a review about how the activities of nociceptive neurons are controlled by higher brain centres and neurochemicals involved in pain transmission. PMID- 16315855 TI - [Age estimation of children in south-western Hungary using the modified Demirjian method]. AB - The biological age of a child, in contrast to his or her chronological age, may be described in terms of skeletal or dental maturity, the development of secondary sex characters etc. These factors can be applied separately or together to assess the degree of physiological maturity of a growing child. The determination of this physiological age and its correlation to the chronological age is of importance within many fields, such as archaeology, forensic medicine, endocrinology, orthodontics etc. Dental maturity was studied on 203 panoramic radiographs, taken between 1985 and 1995, of healthy Hungarian children aged 2.9 to 17.5 years from the south-west Transdanubian part of Hungary. The revised method of Demirjian was used for determination of the maturity score of each individual. For each tooth a score (on a scale A-H) was given according to the radiographic development, from first appearance of calcification to the closure of the apex. This score was then transformed into a self-weighted maturity score for the dental stages (0-19.3) as presented in the paper. The individual scores were then added giving a total maturity score (0-100). This total maturity score represents the biological maturity of the child and may then be compared with the known chronological age. The total maturity scores of the Hungarian population was plotted against the chronological (real) age, and a curve was fitted using the Lowess method of locally weighted least-square method. Our results show that each population needs its own standards. Hungarian boys and girls are approximately 1 year ahead of their French-Canadian counterparts at the age of 6 8. At older ages, the difference decreases. Similar to other studies, we found high individual variance up to 2 years, in tooth development, in the Hungarian population. PMID- 16315856 TI - [Organ transplant patients' complex periodontal treatment]. AB - The number of organ transplant patients has increased in recent years. The Cyclosporin-A (CsA) has been used at least for 20 years to control graft rejections. As many organ transplant patients also take Ca channel blocking medication this might potentiate the CsA associated gingival hyperplasia. The outcome of the comprehensive periodontal treatment was evaluated in 14 kidney and liver transplant patients. If gingival overgrowth showed no marked reduction after thorough professional supra and subgingival plaque control either gingivectomy or flap surgery were indicated. The average age of the organ transplant patients was 38 years. 10 was man and 4 female. The daily dose of CsA ranged between 150-200 mg, and each patient took also Ca channel blocking drugs, 5-20 mg/day. S/RP alone was successful in two cases. Gingivectomy was performed in 10 cases and flap operation was indicated in two cases. Among patients with high compliance and excellent individual oral hygiene no recurrence occurred, while two patients with very low level of oral hygiene showed immediate gingival overgrowth after surgery. It is supposed that dental plaque is an important contributing factor in the development of drug induced gingival overgrowth and meticulous plaque control should be one of the most important treatment modalities in controlling drug-induced gingival changes. PMID- 16315857 TI - [Ten years of experience with the complex periodontal and orthodontic rehabilitation of patients with periodontal diseases]. AB - In periodontal diseases where the loss of periodontal tissue is very extensive, displacement of the teeth may be observed. In the interest of complete healing, periodontal treatment must be followed by orthodontic treatment in such patients. If the basic rules relating to adult patients with a reduced, but healthy periodontium are adhered to, the orthodontics can be carried out with certainty in adults who have been rehabilitated from a periodontal aspect. An improved long term result may be observed in the patients who participate in such complex rehabilitation. Teeth that have been restored to their correct position are not only aesthetically more attractive, but also function more appropriately, are easier to keep clean, and can be treated more simply. PMID- 16315858 TI - [Gardner-syndrome: case report]. AB - The major symptoms of Gardner-syndrome are described by the authors (multiple osteomas, desmoid tumors, colon polyps with malignant tendency) with a case appearing in their department. The importance of early diagnosis is also emphasized. PMID- 16315860 TI - [Report from the commemoration of the 50th anniversary of the founding of the School of Dentistry at Semmelweis University]. PMID- 16315861 TI - CAN: programs & tools to improve your workplace. PMID- 16315862 TI - [Intracellular glutathione (GSH) of monocytes (Mo) directs the development of peritumoral stroma in patients with colon carcinoma]. AB - BACKGROUND: One of the unresolved issues related to immunotherapy for cancers is how to predict the degree of growth of tumor stroma at the periphery of the tumor. We examined the feasibility of predicting stromal growth by measuring the glutathione (GSH) level in monocytes as an indicator of the local redox state of cancer, reflecting anti-tumor immune responses. METHODS: Blood was drawn preoperatively from 21 patients with colorectal cancer. Monocytes were isolated from each blood sample and observed under a fluorescent microscope after fluorescent dye staining. The chromatic responses were rated on a three-grade scale: strongly positive, moderately positive and weakly positive. The monocytes were counted in each grade, and the GSH score was calculated. After surgery, pathology specimens of resected tissue were observed under a microscope to measure the percentage of the surface area of stroma in a given visual field at a magnification of x200. The percentage of stromal surface area was measured for three visual fields (near the deepest area of the tumor) to calculate the stroma score. RESULTS: The mean GSH score was 290.2. The mean stroma score was 60.8. There was a significant correlation between the GSH score and the stroma score (r = 0.421, p < 0.05). CONCLUSION: The monocyte GSH level may serve as a predictor of stromal growth. PMID- 16315863 TI - [Therapeutic potential of antisense (AS) Bcl-2 as a chemosensitizer for patients with gastric and breast carcinoma]. AB - BACKGROUND AND OBJECTIVE: Bcl-2 confers resistance to apoptosis resulting in reduction of the effectiveness of chemotherapy. We examined the effect of antisense (AS) Bcl-2 in combination with anticancer drug for targeting therapy against Bcl-2 in gastric and breast cancer cells. MATERIALS AND METHODS: One human gastric cancer cell line (MKN-45) and three breast cancer cell lines (BT 474, ZR-75-1 and MDA-MB-231) were examined. The effects of antisense Bcl-2 phosphorothioate oligonucleotides (AS ODNs: 18 mer) on chemosensitivity were tested in vitro and in vivo. Chemosensitivity was evaluated by the MTT assay, and the antitumor effect assessed in vivo by the success of xenograft transplantation into nude mice. RESULTS: Treatment with AS Bcl-2 ODNs resulted in sequence specific reduction in protein expression, which was compared to controls. Treatment of MKN-45 cells with AS Bcl-2 increased sensitivity to adriamycin (ADM), cisplatin (CDDP), and paclitaxel (PTX) in vitro and in vivo. Similarly, treatment of BT-474, ZR-75-1, and MDA-MB-231 cells with AS Bcl-2 increased chemosensitivity to ADM, mitomycin C (MMC), PTX, and docetaxel (DOC). This occurred in the setting of increased Bax and cleaved poly (ADP-ribose) polymerase, as well as decreased Bcl-2 and pAkt. AS Bcl-2 ODNs induced splenomegaly in association with increased serum IL-12, expression of CD80, CD83, CD86, and CD27, which was attenuated by methylation of the CpG-motifs of AS Bcl 2, suggesting the involvement of immunomodulatory effect of AS Bcl-2 through pDC and B cell although methylated CpG-failed to negate the increased antitumor effect of AS Bcl-2. CONCLUSION: Targeted therapy against Bcl-2 protein using AS ODNs might enhance the effects of chemotherapy in patients with gastric and breast cancer. PMID- 16315864 TI - [Study of indoleamine 2,3-dioxygenase expression in patients with breast cancer]. AB - We evaluated the clinical significance of indoleamine 2,3-dioxygenase (IDO) in breast cancer. Operative specimens obtained from 30 patients with breast cancer were investigated by semiquantitative RT-PCR with specific primers against IDO. The correlations among IDO expression, clinicopathologic factors and prognosis were studied. The expression of IDO was observed in 100%, both of the cancer specimens and the non-cancer specimens. The IDO expression of the cancer specimens was higher than the non-cancer specimens. The expression of IDO did not correlate to histologic classification, tumor size, lymphatic invasion, venous invasion and lymph nodes metastasis, but correlated to clinical stage and the serum level of immunosuppressive acidic protein (IAP). There were no correlations for a survival rate after surgery between the high IDO level group and the one. The serum IDO levels of cancer patients were higher than that of a healthy volunteer measured by semiquantitative RT-PCR and HPLC. It is suggested that the expression of IDO in breast cancer patients may play a critical role for immunosuppression in those patients. PMID- 16315865 TI - [Evaluation of host immunity and side effects in breast cancer patients treated with adjuvant chemotherapy (FEC therapy)]. AB - BACKGROUND: FEC (5-FU+epirubicin+cyclophosphamide) therapy has been used as adjuvant chemotherapy for breast cancer patients with nodes positive. The aim of this study was to evaluate host immunity and side effects of the FEC therapy. The effect of oral administration of Lentinus edodes mycelia (LEM) was also observed. METHODS: Ten patients were enrolled in this study. The treatment with 5-FU (500 mg/m2), epirubicin (75 mg/m2) and cyclophosphamide (500 mg/m2) was administered every 21 days for 2 cycles, and LEM (9 g/day po) was administered during the 2nd cycle. RESULTS: NK cell activity and the number of white blood cells decreased on the 7th day after the therapy, and they recovered on the 21st day. However, this NK cell activity and the number of white blood cells didn't decrease when the FEC therapy was used with LEM po. CONCLUSIONS: FEC 75 therapy has made some impacts on host immunity, and LEM with the FEC 75 therapy might have prevented host immunity. PMID- 16315866 TI - [Repeated administration of alpha-galactosyl ceramide attenuates of natural killer T cells]. AB - alpha-Galactosyl ceramide (alpha-GalCer) is well known as a specific ligand which can activate natural killer T (NKT) cells. This drug potentially induces anti tumor effect of NKT cells, and clinical trials for alpha-GalCer in cancer patients are ongoing in the world. The aim of this study is to investigate how repeated alpha-GalCer injection affects the activation of NKT cells in mice. The initial administration of alpha-GalCer triggered a rapid production of both IFN gamma and IL-4 in NKT cells and induced subsequent apoptosis in the majority of those cells. Meanwhile, no increase in IFN-gamma production was observed after further injections, and NKT cells maintained the low level secretion of IL-4 since the first injection. After repeated alpha-GalCer administration, activation markers on NKT cells were down-regulated and autologous cytotoxicity against liver cells decreased, suggesting that repeated stimulation attenuates the response of NKT cells to the ligand. These data indicate that prudent discussion is required to determine the dosing interval of alpha-GalCer in clinical applications. A further study is needed for establishing effective methods of sustained NKT cell activation. PMID- 16315867 TI - [Radiation, 5-FU and OK-432: inhibitory effect of IL-10 and TGF-beta]. AB - We investigated the effect of 5-FU and radiation in OK-432-induced cytokine production. Stimulation of peripheral blood mononuclear cells (PBMCs) with OK-432 (1 micro/ml) for 24 h induced Th1-type cytokines (IFN-gamma, TNF-alpha, IL-12, IL 18) as well as IL-10 and TGF-beta. When the PBMCs were stimulated by 5-FU (5 microg/ml) or X-ray (2 Gy) simultaneously with OK-432, production of IL-10 and TGF-beta was significantly inhibited, while no significant change in Th1 cytokine production was observed. Although OK-432 also enhanced the expression of the genes encoding SOCS-1 and SOCS-3, which are negative regulators for cytokine signaling, this was reduced by 5-FU or X-irradiation. Induction of IL-10 and TGF beta by OK-432 was significantly decreased by adding antisense ODN for SOCS-1 or that for SOCS-3. Radiation and 5-FU induce Th1-dominant state by inhibiting the OK-432-induced production of IL-10 and TGF-beta mediated by regulation of SOCS-1 and SOCS-3 expression, and are suggested to increase anti-cancer immunity. PMID- 16315868 TI - [Anti-tumor immunity induced by OK-432-derived DNA]. AB - We have tried to identify the effective components of OK-432, a Streptococcus derived anti-cancer immunotherapeutic agent. In the current study, we investigated the effect of OK-432-derived DNA (OK-DNA) in augmenting anti-cancer immune response. Analysis of OK-DNA with the restriction enzymes Hpa II and Msp I revealed that OK-DNA contained unmethylated CpG motifs. OK-DNA induced Th1-type cytokines, such as IFN-gamma and IL-12, and augmented killer cell activities in vitro on human peripheral blood mononuclear cells, whereas the methylated OK-DNA did not. Cytokines were also produced by OK-DNA-stimulated splenocytes derived from wild-type mice but not from TLR9-deficient mice. In the in vivo study, a peritumoral administration of OK-DNA resulted in a significant inhibition of tumor growth in syngeneic tumor-bearing wild-type and TLR4-deficient mice but not in TLR9-deficient mice. Anti-tumor effect of OK-432 in TLR9-deficient mice was significantly but partially reduced as compared with that in wild-type mice, while the effect of OK-432 was almost completely eliminated in TLR4-deficient mice. These findings suggest that unmethylated CpG-DNA in OK-432 functions as an active component in OK-432-induced anti-cancer immunity via TLR9, at least in part. PMID- 16315869 TI - [Induction of apoptosis in human head and neck cancer cell lines by an active component of OK-432 through p53-independent pathway via toll-like receptor (TLR) 4 signaling]. AB - OK-PSA, an active component of OK-432, induces anti-tumor immunity via Toll-like receptor (TLR) 4/MD-2 complex. In the current study, we evaluated the effect of the OK-PSA on human head and neck cancer cell lines. Twelve cancer cell lines including 7 squamous cell carcinoma (SCC) cell lines and 5 salivary gland cancer (SGC) cell lines were examined. The quantitative real-time PCR analysis revealed that TLR4 mRNA was expressed in all 12 cell lines, and that MD-2 mRNA was expressed in 5 cell lines. OK-PSA stimulation resulted in the activation of NF kappaB in the 4 SCC cell lines which express both TLR4 and MD-2 genes, and in 5 SGC cell lines which express at least TLR4 gene independently of MD-2 expression. In these OK-PSA-responsive cell lines, OK-PSA activated caspase-1, caspase-3 and caspase-8, and induced apoptosis. OK-PSA-induced apoptosis were observed even in a SGC cell line in which p53 is mutated and its function is impaired. These findings strongly suggest that OK-PSA induces apoptosis by the activation of caspases through p53-independent pathway via TLR4 signaling in head and neck cancer cells. PMID- 16315870 TI - [Clinical evaluation of intrapleural or peritoneal repetitive administration of Lentinan and OK-432 for malignant effusion]. AB - LTN and OK-432 combined therapy is effective for controlling Th1/Th2 balance. We tried a repetitive administration of LTN and OK-432 in the pleural or peritoneal cavity for patients with malignant effusion. Of all 11 lesions of the 10 cases, 7 revealed complete remission and 1 revealed partial response. The level of IL-12 (p70) and IFN-gamma in ascites of two gastric cancer patients after the second administration of LTN and OK-432 was much higher than those after the first administration, whereas the level of IL-10 was not suppressed strongly. In 8 lesions that we could confirm complete remission or partial response, 7 lesions were improved after two or three administrations of LTN and OK-432. In conclusion, a repetitive intracavital administration of LTN and OK-432 is effective for malignant effusion. PMID- 16315871 TI - [Usefulness in predicting parameters for the selection of responders who received immunochemotherapy with PSK in patients with colorectal cancer]. AB - It is very important for immunotherapy to release Th2-dominated immunological conditions in patients with malignant diseases. In the present study, we assessed the intracellular cytokine profiles of CD4 positive cells in peripheral blood in patients with colorectal cancer using a flow cytometric analysis and we investigated whether Th2-dominated immunological conditions could be released by PSK. Peripheral blood samples were collected preoperatively from 57 patients with colorectal cancer before and after the oral administration of PSK (3g/day x 1 week). After the PSK treatment, CD4(+)IL-10(+)T-cell percentages decreased significantly, whereas no significant change occurred in proportions of CD4(+)IL 6(+)T-cells. In the after/ before PSK treatment percentages, the ratio of CD4(+)IL-10(+)T-cells were significantly lower in non-recurrent patients compared with recurrent patients, whereas no significant difference was seen in the ratio of CD4(+)IL-6(+) T-cells. These results suggest that the after/before percentage ratio of CD4(+)IL-10(+)T-cells may be useful predicting parameters for the selection of responders. PMID- 16315872 TI - [Two case reports on intra-tumor injection therapy of dendritic cells]. AB - DC (dendritic cells) vaccine therapy against cancer has attracted attention in recent years. However, the existence of the immunosuppressive state in cancer individuals leads to anergy and failure in cytotoxic T cell (CTL) induction and DC migration to the target organ. It has been reported that injected intra-tumor DC is expected to work phagocytosis of the tumor as a localized effect, the consequent CTL induction in the tumor and the regional lymphnodes, resulting in a systemic effect. Two cases reported in this article were performed with intra tumor DC injection therapy by means of EUS (endoscopic ultrasonography) which indicated interesting immunoreaction. PMID- 16315873 TI - [Intratumoral injection of immature dendritic cells (DC) for cancer patients]. AB - We proceeded with DC immunotherapy for 21 cancer patients. Immature dendric cells were injected intratumorally to the 16 patients, and three good and effective cases were obtained: case 1: A 69-year-old male patient with papilla-vater carcinoma, case 2: A 49-year-old female patient with gastric cancer, case 3: A 66 year-old male patient with malignant melanoma. PMID- 16315874 TI - [The effect of cimetidine mainly increases CD4+ cells of peripheral blood T lymphocytes]. AB - Cimetidine, one of the most popular histamine-2 receptor antagonists, has been reported to improve survival in gastrointestinal cancer patients and to activate cell-mediated immune response in surgical patients. NKT cells are a population of T cells that share characteristics with natural killer cells, and their main functions are production of immunoregulatory cytokines and cytolytic activities. In this study, we aimed to investigate the effect of cimetidine on the cell mediated immunoresponse. Six healthy adult volunteers were given 800 mg of cimetidine per day orally, and their blood samples were taken prior to and at days 1, 3, 5, and 7 days post-administration of cimetidine. Leukocyte counts and differentials were obtained by the conventional hemogram, and the leukocyte subsets were analyzed by flow cytometry. Cimetidine administration caused leukocytosis, dependent on the increase of neutrophils, as well as of the CD3 positive T lymphocytes, and the subset of CD4-positive cells among them. On the other hand, the NK cell subpopulation was decreased, and the NKT cell subpopulation was not affected. The present results suggest that cimetidine is a modulator of the cellular immunity, and may be used as the activator of the tumor specific immunoresponse. PMID- 16315875 TI - [An interesting change of E-selectin in cimetidine administration during anticancer drug use]. AB - E-selectin is an adhesion molecule developed as an important material for hematogenous metastasis of cancer cells on vascular endothelial cells. It is expected that if we can restrain a manifestation of E-selectin then hematogenous metastasis can be restrained. We divided gastric cancer and the colorectal cancer patients, who performed chemotherapy, into two groups of cimetidine administrated group and a non-administration group, and reviewed whether cimetidine inhibited an expression of E-selectin on vascular endothelial cells by measuring E-selectin in plasma. We experienced one example that showed an interesting change of E selectin and the quantity of E-selectin in plasma fell during the cimetidine dosage. However, we report that E-selectin has risen after the cimetidine dosage was cancelled in the cimetidine administrated group. PMID- 16315876 TI - [Fusion vaccine therapy by IL-2-gene-transduced dendritic cells and tumor cells]. AB - We evaluated the usefulness of fusion vaccine prepared from IL-2-gene-transduced splenic dendritic cells (DCs) and fibrosarcoma tumor cells (QRsP) in treating of lung metastasis. The IL-2 or LacZ gene was transferred into spleen-derived DCs using an adenoviral vector. Irradiated QRsP tumor cells were fused with IL-2 gene transduced DCs (fusion/IL-2) or LacZ gene transduced DCs (fusion/LacZ) by polyethyleneglycol. These fusion cells expressed major histocompatibility complex (MHC) class I and MHC class II, CD86, CD11c and CD8alpha. IFN-gamma and cytotoxic T lymphocyte (CTL) activity of splenic lymphocytes in mice vaccinated with fusion cells increased significantly as compared with those of DC or tumor cells vaccinated mice. CTL levels in fusion/IL-2-vaccinated mice were higher than that in fusion/LacZ-vaccinated mice. The number of lung metastasis in the fusion/IL-2 or fusion/LacZ-vaccineatd mice was significantly lower than that in mice vaccinated with DCs, tumor or PBS. The introduction of the IL-2 gene into fusion cells produced more potent therapeutic effects. Our results suggest that the fusion cells prepared from IL-2 gene transduced spleen derived DCs and tumor cells have the ability to induce therapeutic effect against lung metastasis. PMID- 16315877 TI - [The effects of direct hemoperfusion using the filtration column filled with the adsorption fiber for immunosuppressive substances on cell-mediated immunity in tumor-bearing rats]. AB - The patients with advanced cancer often lose their anti-tumor immune responses due to the increase of some immunosuppressive substances in the blood, such as cytokines and proteins derived from cancer cells or immune cells. We developed the adsorption fiber in transforming growth factor (TGF)-beta for the treatment to remove immunosuppressive substances and investigated the effects of direct hemoperfusion using the filtration column filled with this adsorption fiber in tumor-bearing rats. On day 0, KDH-8 tumor cells (1 x 10(6) cells/rat) were implanted subcutaneously into the back of WKAH/Hkm rats. On day 21 after tumor implantation, the rats underwent the direct hemoperfusion with this filtration column for 60 minutes. On day 28, the rats were sacrificed and the natural killer (NK) activities of their spleen cells were examined. As a result, the rats that underwent this treatment showed a significant increase in their NK activities compared with those of rats who underwent direct hemoperfusion with an empty column or had no treatment. Therefore, we indicated the possibility of a new immunotherapy technique against cancer using a direct hemoperfusion column filled with an adsorption fiber for immunosuppressive substances. PMID- 16315878 TI - [Soothing effect of Ganoderma lucidum antlered form on cyclophosphamide-induced adverse reaction]. AB - The immunological functions of Ganoderma lucidum antlered form (AF) (Rokkaku Reishi in Japanese), a variant type of Ganoderma lucidum, were investigated in C57BL/6 mice treated with cyclophosphamide (CY). Ganoderma lucidum AF alleviated CY-induced decrease in body weight and abnormal increase in blood neutrophil level, when the mice were fed a diet containing 2.5% Ganoderma lucidum AF starting one week before CY treatment (150 mg/kg, ip). The recovery of CD8+ and NK1.1+ cells in the spleen was accelerated in Ganoderma lucidum AF group compared to the control group. Ganoderma lucidum AF also both alleviated CY-induced splenic lymphopenia and suppressed the abnormal increase in splenocytes 7 days after CY treatment. These results suggest that ingestion of Ganoderma lucidum AF is beneficial for improvement of quality of life reduced by anti-cancer chemotherapeutic drugs such as CY. PMID- 16315879 TI - [Host immunity in colorectal cancer patients treated with TS-1 plus CPT-11]. AB - TS-1/CPT-11 treatment for the patients with advanced colorectal cancer is expected to be equal to FOLFOX and FOLFIRI regimen. The aim of this study was to investigate the relationship between host immunity and this regimen. Four patients were enrolled in this study. Host immunity was investigated before and after chemotherapy. PHA response, serum IAP level and the productions of any cytokines did not change significantly before and after the chemotherapy. Although these changes were not significant: both Th1/Th2 ratio and NK cells ratio were decreased, and the ratio of suppressor T cells was increased. These results suggest that TS-1/CPT-11 regimen may influence host immunity a little bit worse. PMID- 16315880 TI - [Radiofrequency ablation and percutaneous ethanol injection in patients with unresectable hepatocellular carcinoma--a comparative study using Japan Integrated Staging Score (JIS score) system]. AB - The patients were classified according to the Japan Integrated Staging (JIS) score and a comparison examination of the long-term therapeutic response of 149 unresectable hepatocellular carcinomas: 90 of them given percutaneous ethanol injection (PEI), and 59 of them given radiofrequency ablation (RFA) who were diagnosed with 3 cm or less in diameters of tumor and less than three nodules, or 5 cm or less single nodule, was carried out in retrospective. To all the tumors exceeding 3 cm in diameter, we added transcatheter arterial embolization (TAE) prior to PEI or RFA. Ninety four percent of PEI cases and 25% of RFA cases received a combination therapy with TAE, respectively. The three-year survivals were 86%, 76%, and 56% in PEI and 96%, 83%, and 68% in RFA for JIS-0, 1, and 2, respectively. The five-year survivals were 69% and 53% in PEI and 95% and 83% in RFA for JIS-0 and 1, respectively. The significant difference in the probability of survival was not confirmed among both treatments. We conclude that the combination therapy, such as combined use of TAE, PEI and RFA as local treatment, can expect almost equivalent effectiveness for unresectable hepatocellular carcinoma. PMID- 16315881 TI - [Radiofrequency ablation for hepatocellular carcinoma and liver metastases]. AB - We evaluated the efficacy of radiofrequency ablation (RFA) therapy for 29 patients with 36 hepatocellular carcinoma (HCC) nodules and 16 patients with 38 metastatic hepatic nodules. The mean tumor size was 26.4 mm. The primary lesions of patients with metastatic liver tumors were 9 colon cancer, 2 rectal cancer, 2 breast cancer, 2 gastric cancer, and 1 esophageal cancer. All nodules were treated using a Cool-tip RFA system. US-guided RFA was performed for 44 nodules, CT-guided RFA for 24 nodules, and intra-operative US-guided RFA for 6 nodules. In a mean observation period of 13.5 months, the mean complete ablation rate and the mean distant recurrence rate were 83.3% and 30.6% for HCC and 65.8% and 31.6% for metastatic nodules, respectively. The mean complete ablation rate of HCC was significantly higher than that of metastatic nodules (p < 0.05). The mean complete ablation rates of both HCC and metastatic hepatic nodules 3 cm or smaller in diameter were significantly higher than those of both tumors larger than 3 cm in diameter (p < 0.05). The mean distant recurrence rate of HCC in patients who have multiple nodules was 62.5% and it was significantly higher than that in patients who have a single nodule (28.6%) (p < 0.05). The mean complete ablation rate of metastatic nodules by intra-operative US guided RFA was 100% and it was statistically higher than that by other image guided RFA (p < 0.05). PMID- 16315882 TI - [Endoscopic radiofrequency ablation for liver metastasis of colorectal cancer]. AB - The application of radiofrequency ablation (RFA) for liver metastasis of colorectal cancer has not yet acquired an established status in clinical cancer therapy research. Removing as much tumor tissue as possible is desirable, but some cases do not allow optimal surgical ablation due to general condition of the patient and tumor status. We introduced endoscopic RFA for liver cancer in 2003, and have applied the procedure to 6 cases with H1 or H2 liver metastases of colorectal cancer to which surgical ablation could not be applied due to the poor general health of patients. Mean tumor diameter was 22.9 mm, and mean number of tumors per patient was 1.2. Tumor location was: S4, n = 2; S5, n = 1; S4, n = 1; S7, n = 2; and S8, n = 1. Mean frequency of session was 3.0. No complications occurred in any cases, and no reoperations were required. Although no recurrence of tumors in the vicinity of ablation was observed, 2 cases of each lung metastasis and intrahepatic recurrence were identified. Intrahepatic recurrence underwent hepatic arterial infusion (HAI) chemotherapy for simultaneous metastatic hepatic tumors (H2) prior to RFA, and relapses occurred in the metastatic focus where the efficacy of HAI was observed. At this point, 2 deaths were reported, 1 each from cancer and other diseases, and mean duration of survival after the procedure was 451.2 days. These results indicate that endoscopic RFA with good local control should be an available treatment for cases involving colorectal cancer with metastasis to the liver in which surgical ablation is difficult to apply. PMID- 16315883 TI - [A clinical study of endoscopic local coagulation therapy for hepatocellular carcinoma (HCC)]. AB - Eighteen subjects with cases of HCC who underwent endoscopic local coagulation therapy at Hiroshima City Hospital between 1998 and 2004 were studied and compared with 6 cases of HCC patients who underwent laparoscopic partial hepatectomy during the same period. The subjects composed of 10 cases of laparoscopic microwave coagulation therapy (L-MCT), 5 cases of laparoscopic radio frequency ablation therapy (L-RFA), and 3 cases of thoracoscopic microwave coagulation therapy (T-MCT). The operation time was 114 minutes for L-MCT and 92 minutes for L-RFA, both of which were significantly shorter than 208 minutes for resection cases. No complications were developed in any of the cases and the postoperative hospitalization period of the cases was 13.1 days, 8.2 days, and 13.0 days, respectively. Although each case of local recurrence was observed both in L-MCT and L-RFA groups, one case displayed observation difficulty from the liver surface and the other required a daughter nodule. The three-year survival rate was 71.4%, while the five-year survival rate was 53.6%. Endoscopic local coagulation therapy is not too invasive and useful for hepatocellular carcinoma in which percutaneous RFA is difficult. However, it is well indicated when the scope is evident with observation being feasible from the liver surface. PMID- 16315884 TI - [A new approach to local ablation therapy for hepatocellular carcinoma--hand assisted laparoscopic radio-frequency ablation]. AB - We have developed a radio-frequency ablation using hand-assisted laparoscopic surgery (HALS-RFA) for hepatocellular carcinoma, and it applied for 10 patients who were difficult to be treated by common laparoscopic surgery (LS) from November 2001. The reasons for selection of HALS-RFA were severe adhesions in 3 patients, anatomical location at the hepatic dome in 3 patients, lesions adjacent to other organs or vasculatures in 3 patients, and co-operation for another organ in 1 patient. We compared efficacies of HALS to other ablation therapeutic techniques with LS and open surgery (OS). The average amount of bleeding was 85 ml in HALS, 14 ml in LS and 319 ml in OS. The amount of HALS was lesser than that of OS, but not significant. Post operative maximal CRP level was 3.2 mg/dl, 4.9 mg/dl and 10.5 mg/dl in HALS, LS, and OS, respectively, with a statistical significance between HALS and OS. Complications occurred in 2 out of 10 cases (20%) in HALS, 2 out of 62 cases (3.2%) in LS and 3 out of 9 cases (33%) in OS. Recurrence in the treated site was encountered in 1 out of 10 cases (10%) in HALS, 2 out of 62 cases (3.2%) in LS and none (0%) of OS. Based on these findings, we conclude that HALS-RFA seems to be a minimally invasive and highly effective treatment. PMID- 16315885 TI - [Evaluation of biliary stenting therapy for malignant biliary stricture at our hospital]. AB - The clinical results of biliary stenting therapy for non-resectable malignant biliary stricture were assessed for patency periods and survival periods in consideration of the types of stents and disease. The subjects were 51 patients with non-resectable malignant biliary stricture due to disease of the pancreas and biliary tract treated at our hospital between April 2001 and March 2005. No significant differences were observed in the patency periods and survival periods among the various types of stents. In regard to the type of disease, the survival period for biliary tract cancer was longer (616 +/- 246 days) than those for other diseases. There was no difference in the number of times of stenting among the various stents, and jaundice was effectively reduced. No patients in our series died of failure to reduce jaundice. We therefore considered it necessary to choose a proper stent indwelling route and a stent type according to stricture site, disease type and PS (performance status) at the time of stenting. The usefulness of the biliary stenting therapy for non-resectable malignant biliary stricture was suggested. PMID- 16315886 TI - [The best choice of biliary stents in hilar distal strictures due to inoperable malignacies]. AB - Recently, the treatment using biliary stents play an important role to improve the quality of life (QOL) in patients with inoperable malignant biliary obstruction. Long-term patency is preferable, but the convenience of treatment for stent obstruction should also be considered. In this study, we investigated which stent is currently the most appropriate in hilar and extrahepatic bile duct obstruction. In hilar obstruction, metallic stents (MS) had significantly longer patency than plastic tube stents (TS). As for extrahepatic bile duct obstruction, covered MS (CMS) and MS had significantly longer patency than TS. On the other hand, many difficulties occurred when the MS was obstructed. In some of these patients, it was difficult to place the internal drainage again and their QOL was severely impaired. CMS was more convenient because it could be removed when it was obstructed. These results may suggest that MS should be used carefully in the hilar obstruction and that CMS is the best for the treatment of extrahepatic bile duct obstruction. PMID- 16315887 TI - [Low-dose cisplatin plus 5-fluorouracil with radiotherapy for unresectable upper and hilar bile duct carcinoma]. AB - We report herein the value of a combined treatment modality for bile duct cancer (intraluminal brachytherapy: ILBT+extrabeam radiation therapy+chemotherapy+expandable metallic biliary stent), employed for cases in which excision of the upper and the hilar bile duct cancer was not possible. The combined treatment modality group did not attain a two-year survival rate according to the Kaplan Meier method. They were not able to surpass the treatment results of the excision group which ended in radical cure level of C. Moreover, with the combined modality treatment, the addition of radiation therapy significantly improved the cumulative survival rate. In conclusion, the first choice of treatment for the upper and the hilar bile duct cancer is excision, and it is necessary to thoroughly examine the possibility of excision. Additionally, to surpass the treatment results of the excision group that ended in a radical cure level of C, it is necessary to include radiation therapy and chemotherapy using new anti-cancer drugs in a combined treatment modality. PMID- 16315888 TI - [Intra-arterial infusion chemotherapy for advanced cancer--40 years of experience]. AB - During the period of 1990-2005, 701 patients with the hepatocellular carcinoma were treated with intra-arterial infusion of 5-FU and epi-adriamycin with or without Lipiodol chemoembolization employing an implantable infusion port system. In 70% of the patients treated, an objective response was observed with marked regression of tumor and decrease in tumor marker (AFP and PIVKA-II). Also 1,091 patients with the metastatic liver cancer of colon, rectum, stomach and pancreas were treated with the same procedure employing 5-FU, mitomycin C, adriamycin, or epi-adriamycin. In more than 80% of the patients treated, an objective response was observed with marked regression of tumor and decrease in tumor marker (CEA, CA19-9, TPA, DUPAN-2, SPan-1). Intra-arterial infusion chemotherapy employing an implantable port system also proved to be a promising treatment modality for most of the intractable head and neck cancer, breast cancer and a few of the pancreas cancer. PMID- 16315889 TI - [Clinical outcome of the use of self-expandable metallic stents (SEMS) for palliation of patients with malignant upper GI obstruction caused by incurable gastric cancer]. AB - Ten patients with malignant upper gastrointestinal obstruction caused by incurable gastric cancer underwent an endoscopic placement of self-expandable metallic stents for palliation. There were 6 male and 4 female patients aged 39 98 (median, 65.5). The site of obstruction was the cardia in 2, the pylorus in 4, the duodenum in 1, and surgical anastomosis in 3 patients. All the patients were unable to take food orally. Stent deployment was technically successful in all patients. After the stent insertion, 80% of patients resumed solid diet. The median survival time after the insertion was 129 days. The median length of home stay after the procedure was 54.5 days. Although chemotherapy was added in 5 patients after the insertion, there were no significant differences either in survival time or in home stay between the patients with and without chemotherapy. No procedure-related mortality was observed. Mediastinitis occurred in 1 patient, and the remaining 9 patients had no serious complications. Stent migration was observed in 2 patients, and stent occlusion due to tumor ingrowth occurred in 4 patients. PMID- 16315890 TI - [Palliation of malignant bile duct obstruction with metallic stent-clinical effectiveness in aged patients]. AB - To evaluate the clinical effectiveness of metallic stent in the palliation of malignant bile duct obstruction in aged patients, 30 patients over 65 years of age with malignant biliary obstruction were investigated retrospectively. Overall survival duration after the stent placement was 13-1,275 (mean: 278, median: 169) days. The period of tube-free on the outpatient basis after stent insertion was 0 1,162 (mean: 192, median: 121) days. The estimated cost savings by eliminating hospitalization was greater than the stent cost. Four patients survived over 18 months despite their advanced clinical stages. It seems difficult to develop guidelines for the indication of stent placement in the treatment of malignant bile duct obstruction for aged patients. PMID- 16315891 TI - [Evaluation of the endoprosthesis by metallic stent and tube stent for malignant biliary stenosis]. AB - We evaluated the effect of biliary endoprotheses for 20 malignant stenosis patients by an expandable metallic stent and hydrophilic heparinized tube (H-PSD) connected to an implantable port (IP), which reduces bacterial adherence. Group A consisted of 6 patients of cholangiocarcinoma who underwent hepatic arterial infusion chemotherapy associated with radiotherapy. Groups B and C consisted of 8 and 6 patients of stage IVa and IVb pancreatic carcinoma, respectively, who underwent hepatic and splenic arterial infusion chemotherapy following transcatheter peripancreatic arterial embolization. The 50% patent time was 12 months, 6 months and 7 months in groups A, B and C and the 50% overall survival time was 16 months, 23 months and 13 months, respectively. There were two complications, 1 case of infection around the IP in which the IP was withdrawn, and 3 cases of cholangitis in which we had easy access to the bile duct via IP. This technique appears to offer significant benefit in selecting patients with this type of biliary obstruction. PMID- 16315892 TI - [A change of tumor PyNPase level at intraperitoneal and intravenous administrations of paclitaxel]. AB - A level of PyNPase activity was measured after intraperitoneal (ip) and intravenous (i.v.) administrations of paclitaxel on the animal model. Nude mice received the subcutaneous implantation of WiDr cells. About 3 weeks later, the ip and i.v. administrations of paclitaxel were performed 2 times at 20 mg/kg and 15 mg/kg, respectively. About 1 week later, the mice were sacrificed. The level of PyNPase activity was measured by the ELISA method. The level of PyNPase of ip and i.v. was higher than that of the control group, but the level of PyNPase revealed no significant difference between ip and i.v.. This result suggested that intraperitoneal administration of paclitaxel enhanced an efficiency of 5'-DFUR and capecitabine as much as intravenous administration of paclitaxel. PMID- 16315893 TI - [Effect of intraperitoneal chemotherapy on experimental peritoneal dissemination of gastric cancer]. AB - In vitro chemosensitivity test using a collagen-gel method was done on 165 primary gastric cancers. All of 5-FU, CBDCA, CDDP and docetaxel showed a high sensitivity. The effects of per oral (po) administration of TS-1, a combination of po TS-1 and intraperitoneal (ip) administration of CDDP, ip 5-FU and ip docetaxel, were evaluated in athymic mice bearing peritoneal dissemination of a gastric cancer cell line (MKN-45-P that shows a high rate of metastasis to the peritoneal cavity of nude mice). Nude mice were inoculated by ip with 10(7) MKN 45-P cells. No survival benefit was obtained after po administration of TS-1 (12 mg/kg) alone or ip CDDP alone. However, a combination of po TS-1 (8 mg/kg x 10 days, from day 3) and ip CDDP (3.5 mg/kg, day 6 and 13) showed a significant survival improvement than that of po TS-1 or ip CDDP treatment alone. ip administration of 30 mg/kg (3 times/week x 3 weeks) or 15 mg/kg (6 times/week x 3 weeks) of 5-FU significantly improved the survival of mice bearing MKN-45-P. 5-FU concentration of ascites after ip administration of 30 mg/kg of 5-FU was 600-fold higher than po administration of 12 mg/kg of TS-1 at peak level. ip injections of docetaxel of 8 mg/kg, and 2 mg/kg improved the survival of 4 and 1 mice, respectively, and they were tumor-free on day 90. Survival of mice treated with ip injection of CBDCA (100 mg/kg, on day 3, or 50 mg/ kg on day 3 and 10) was significantly better than the control group. These results suggest the potential of po TS-1 + ip CDDP, ip 5-FU, ip docetaxel and ip CBDCA administration for the treatment of peritoneal dissemination of gastric cancer. PMID- 16315894 TI - [Safety and efficacy of chemotherapy using TS-1 followed by paclitaxel for advanced or recurrent gastric cancer with peritoneal dissemination]. AB - There has been no standard treatment for gastric cancer with peritoneal dissemination. We have used TS-1 followed by paclitaxel for advanced or recurrent gastric cancer patients with peritoneal dissemination since January 2002. Twenty three patients were enrolled to our prospective study and 19 of 23 patients completed the protocol. There were less severe adverse events concerning paclitaxel despite of the second line therapy of TS-1, and 80 percent of all therapeutic courses was at an outpatient clinic. The median time to progression was 199 days. The median survival time was 363 days in all the enrolled patients, and was 436 days in 19 patients who completed the protocol. Chemotherapy using TS 1 followed by paclitaxel is considered to be safe and effective for gastric cancer with peritoneal dissemination. PMID- 16315895 TI - [Clinical significance of washing cytology in patients with advanced gastric cancer]. AB - Since 1988, we have performed intraoperative peritoneal washing cytology (CY) of the Douglas pouch in 1,255 patients (pts) with gastric cancer to diagnose latent peritoneal dissemination. Patients were classified as follows: 914 pts in P0, CY0; 100 pts in P0, CY1; 90 pts in P1; 53 pts in P2; and 98 pts in P3. Usually Papanicolaou's staining method was used and immunocytochemical staining of CEA and MOC-31 was carried out to make a definitive diagnosis. Prognostic factors were determined in 638 pts without liver metastasis by multivariate analysis. Curability, depth of tumor invasion, lymph node metastases, histological type and CY were independent prognostic factors. The survival of P0, CY1 pts who had undergone curative surgery was better than those with non-curative surgery. In conclusion, CY can serve as an independent predictor of prognosis, so immunocytochemical staining is important. P0, CY1 pts without residual cancer should be treated with multimodal therapy. PMID- 16315896 TI - [Microwave ablation for multiple bilobar liver tumors from colorectal cancer]. AB - AIM: The aim of study was to evaluate the effectiveness of microwave ablation for multiple bilobar liver tumors from colorectal cancer. METHODS: Data from the patients undergoing a hepatic resection with or without microwave ablation at our department were analyzed retrospectively. Survival was estimated using the Kaplan Meier method and compared using the logrank test. RESULTS: Seventeen percent of patients with combined resection/ablation at sites of initial recurrence at the liver and an extra hepatic metastasis in 11%. The difference between patients with combined resection/ablation and with resection was not significant (p = 0.84). Recurrence occurred near the resection or ablation line in 12.5% within combined resection/ablation. No significant differences were found for overall survivals: disease-free survival between resection/ablation group and with resection alone. CONCLUSION: Microwave ablation plus hepatic resection is suggested to be equally effective as hepatic resection, and it expanded indications for surgery to treat multiple bilobar liver tumors from colorectal cancer. PMID- 16315897 TI - [Cryotherapy of the resection edge after hepatectomy of colorectal liver metastases]. AB - Obtaining a one-centimeter negative margin is an important factor in preventing disease recurrence after surgery for hepatic tumors. Cryotherapy of the resected edge has been used to achieve optimal margin clearance in cases in which the alternative would be an extended high-risk liver resection. As a concrete method, cryotherapy was delivered with a liquid nitrogen based compact system (CRY-AC, Brymill Co., USA). The resection edge with involved or inadequate resection margins was ablated directly by using the flat probe for 3 minutes per 1 place. Between 2002 and present, a total of 14 patients with colorectal liver metastases underwent edge cryotherapy. Although there was no hemorrhage from the stump, postoperative leak of the bile and stump recurrence were recognized in each patient. Since cryotherapy has features that make the vascular difficult to be damaged, the complication was not recognized in the patient with exposed vascular in the resected edge. By extending the follow-up period, we want to examine whether the edge recurrence could be controlled or not. PMID- 16315898 TI - [A case of complex endometrial hyperplasia with atypia treated by microwave endometrial ablation]. AB - Complex endometrial hyperplasia with atypia (CEHA) often precedes an endometrial carcinoma. A nulli-gradiva woman at high risk due to various complications underwent microwave endometrial ablation (MEA) at a frequency of 2.45 GHz for treatment of CEHA as an alternative to hysterectomy. The endometrium near the internal orifice was conserved to avoid hematometra. Endometrial biopsy specimen from the neighborhood of the internal orifice did not show any signs of endometrial hyperplasia postoperatively. Two years after the operation, endometrial biopsy revealed recurrence. The second MEA was performed. MRI one month after the second operation revealed that the uterine lining was completely replaced by avascular area without signs of regrowing endometrium or endocevix. Eighteen months have passed without recurrence. PMID- 16315899 TI - [Antitumor effects of intratumoral injection of Basidiomycetes preparations]. AB - The antitumor effects of Basidiomycetes preparations in an experimental mouse model, the "double grafted tumor system" were analyzed. Some BRMs prevented metastases by utilizing the anti-tumor immunological cascade reactions, which activates macrophages in the body. The following Basidiomycetes preparations were analyzed: PSK was a hot water extract of cultured mycelia from Coliolus versicolor and a protein bound beta-glucan. Matsumax was extracted from mycelia of Tricholoma matsutake and was a protein bound (38%) a-glucan. The Agaricus preparation was extracted from fruit bodies of Agaricus blazei and a protein bound (17%) a-glucan, beta-glucan. Himematsutake preparation was extracted from fruit body of Agaricus blazei (Himematsutake) and a protein bound (5%) glucan. Lentinan was purified from fruit bodies of Lentinus edodes and is a purified beta glucan. PSK cured both primary and metastatic tumors in the double grafted tumor system. Lentinan inhibited the growth of neither primary nor metastatic tumors. Matsumax and Agaricus preparation cured primary tumor and inhibited the growth of metastatic tumor. Himematsutake preparation inhibited the growth of primary tumor. Immunosuppresive acidic protein (IAP) is produced by activated mactrophates. The PSK, Matsumax, Agaricus preparation and Himematsutake preparation induced IAP but Lentinan did not. PMID- 16315900 TI - [Local ablation therapy for hepatocellular carcinoma (HCC) on the liver surface: radio-frequency ablation aimed at tumor marginal pre-ablation under endoscopic surgery]. AB - RFA for the hepatocellular carcinoma localized on the surface of the liver tends to have some complications such as bleeding, an ejection of tumor and a heat injury to other internal organs even if percutaneous RFA seemed to be done easily. Therefore, we should first choose the RFA treatment under endoscopic (either laparoscope or thoracoscope) surgery for the hepatocellular carcinoma localized on the surface of the liver. Moreover, a direct central puncture should be avoided from the viewpoint of securing a margin, prevention of bleeding and rise in the intratumorale pressure. Now, we selected the unique operation method of RFA: First, the tumor is confirmed under the endoscope, and the tumor range is marked with the endoscopic echo. Second, several times of RFA applied to the tumor surroundings are done, and the margin is secured with avoiding a direct central puncture. If tumor diameter is over 2.5 cm, central ablation of the tumor is considered to be necessary, we can directly puncture the center of the tumor without bleeding since the tumor already has the congelation by surrounding heat effect. We have done RFA by this way for 29 patients with HCC since April 1st, 2004. The complications such as a heat injury to the neighboring organ could be well prevented. An enough margin of ablation about 1 cm around the tumor was confirmed by the postoperative CT image. There was no local recurrence during the average observation period of 290 days, and a severe post operative complication has not occurred. The average of hospitalized period after the operation was about 10 days. Therefore, pre-surrounding ablation preceding central puncture under the endosope for hepatocellular carcinoma on the liver surface is a feasible technique. PMID- 16315901 TI - [Utility of convex echo probe in laparoscopic radio frequency ablation therapy for hepatocellular carcinoma]. AB - We have examined the utility of the convex echo probe, which has the fine gutter of a puncture needle in laparoscopic radio frequency ablation therapy. When we use a flexible linear echo probe in RFA treatment, we have to puncture tumor with the hand piece in free hand. But it is difficult to treat in the case of HCC which is located in S1 and the lower area of S5 and S6 because we have a narrow space where colon, duodenum and netz are close for safe and exact puncturing of the tumor. We used a convex echo probe in RFA to the above mentioned area of the liver. We punctured with the hand piece exactly and easily without preliminary puncturing of the tumor. So we can perform RFA treatment successfully and safely by a choice of an appropriate echo probe. PMID- 16315902 TI - [Experience of radiofrequency ablation therapy for hepatocellular carcinoma]. AB - There are various therapeutic options for hepatocellular carcinoma. Radiofrequency ablation (RFA) was introduced to Japan in 1998, and has become popular in percutaneous local treatment for HCC as a treatment with the advantage of both percutaneous ethanol injection therapy and percutaneous microwave coagulation therapy. In this study, we investigated the efficacy and complication of RFA for HCC. Seventeen patients underwent percutaneous or open RFA with a Cool tip needle (Radionics Co. Ltd) from April 2001 to May 2005. All tumors were solitary and the average diameter of 17 tumors was 2.24 cm. Fifteen tumors were completely ablated, but two weren't. Local recurrence occurred in only one patient and the rate of local recurrence was 6.7%. Mild complication occurred in some patients, but critical complication did not occur in any patients. In this study, it was considered that RFA could be performed safely and was a good treatment for HCC with high efficacy. After investigating the long-term results and indication of RFA, it was suggested that RFA might be reestablished as an effective treatment for HCC. PMID- 16315903 TI - [Results of local ablation therapy for liver metastases from colorectal cancer using radiofrequency ablation and microwave coagulation therapy (RFA/MCT)]. AB - In addition to hepatectomy and chemotherapy with intra-arterial infusion, local ablation therapy using RFA and MCT has been used for treatment of liver metastases derived from colorectal cancer. We investigated the results of local ablation therapy in such cases conducted at our department. SUBJECTS AND METHODS: This study was conducted in 9 out of 21 patients with liver metastases derived from colorectal cancer, who underwent local ablation therapy with RFA from September 2001. As adjuvant therapy, the divided administration of low-doses CDDP/FU was conducted in all cases. RESULTS: RFA and MCT were used in 9 cases (25 lesions) and in 15 cases (82 lesions), respectively. The three cases of RFA were combined with MCT. The severity of liver metastasis was H1 and H2 in one case each and H3 in seven cases. The tumor diameter was 1.0-4.7 cm (mean, 2.7 cm) and patients underwent cauterization from 1-6 times (mean, 2.4 times). Although there were no critical complications, pyrexia and increased AST/ALT were found. The three-year survival rate was 50%, and thus favorable prognoses were obtained. CONCLUSION: There were no notable complications associated with local ablation therapy for liver metastases derived from colorectal cancer, and this method was thus indicated to be effective for local control in cases in which hepatectomy is impossible. PMID- 16315904 TI - [Radio-frequency ablation therapy for metastatic liver tumors from colorectal carcinoma]. AB - Radio-frequency ablation therapy (RFA) as a treatment for metastatic liver tumors from colorectal carcinoma was examined. METHODS: Ten patients with a total of 30 liver metastases from colorectal carcinoma were treated using a Cool-tip RF system from March 2003 to December 2004. RESULTS: Patients had a mean age of 69.8 years and the mean diameter of the metastatic lesions was 29.5 mm (range, 5-82). Two patients had received RFA therapy 2 times, and another 2 patients had received 3 times. Critical complications were not seen, though 5 therapies were performed using CT-guided trans-pulmonary puncture. The rate of partial recurrence was 23.1% and the average observation period was 14.8 months. The partial recurrence had occurred within the mean period of 6.2 months. Although after multimodal therapy was given, it is suggested that repeated RFA for the liver metastasis would improve survival rates. CONCLUSION: RFA is a safe and effective treatment for metastatic liver tumors from colorectal carcinoma as multimodal therapy. PMID- 16315905 TI - [Radiofrequency ablation (RFA) in colorectal cancer with hepatic metastases]. AB - We considered the appropriateness of RFA, which was performed in three cases of colorectal cancer with hepatic metastases accompanied by liver cirrhosis. Case 1 involved a patient with sigmoid colon cancer ss, n1 (+) with severe hepatic dysfunction and synchronous hepatic metastases (S5, S6, S8) in which RFA was performed. After 1 year and 6 months, recurrence (S3, S4) was detected in the residual liver, and the patient is currently undergoing the IFL (CPT-11/5 FU/Leucovorin) treatment. In case 2, following a partial hepatic resection, RFA was performed for cecal cancer ss, n2(+) with synchronous hepatic metastases (S5, S6, S8). After 11 months, recurrence (S5, S6, S7) occurred in residual liver and again RFA was performed following a partial hepatic resection. Lung metastases have occurred and currently IFL (CPT-11/5-FU/Leucovorin) and WHF treatments are underway. In case 3, 4 years and 8 months after cancer of the descending colon ss, n1 (+), RFA was performed on asynchronous hepatic metastases (S5, S7, S8). The patient died of peritonitis carcinomatosa one year after RFA. In all three cases, metastases were identified by dynamic CT as low density masses with no blood flow. Necrosis in all three metastases and local control had been achieved. There were no severe complications. Under the current conditions, local coagulation methods including RFA are appropriate in those cases in which resection are not possible such as multiple metastases with severe hepatic dysfunction, etc. PMID- 16315906 TI - [Radiofrequency ablation of liver metastasis from breast cancer]. AB - As the hepatic metastasis from breast cancer has a tendency to have an extrahepatic lesion, systemic therapy therefore becomes acclimatization. However, local therapy is regarded as one of the choices if there is no extrahepatic lesion. We present three cases of liver metastasis from the breast treated by radiofrequency ablation (RFA). Case 1: A 65-year-old woman was treated by left mastectomy in 1997. Radiation exposure was performed for lung metastasis, and a weekly paclitaxel therapy was administered in 2001. We performed RFA percutaneously for liver metastasis of 2.8 cm in 2002. The aggravation spread to the lung lesion and she died after RFA within one year. The liver metastasis finally enlarged to 4 cm in size. Case 2: A 36-year-old woman was treated by left mastectomy (Stage IIIa), and was followed by chemotherapy in 2000. We performed RFA for metastasis of 2 cm of liver (S7) percutaneously in 2001, and didn't recognize a recurrence to date for 3 years and 8 months. Case 3: A 43-year-old woman was treated by left mastectomy (Stage IIIa), and followed by chemotherapy in 2003. We performed RFA for a liver metastasis of 3.5 x 4 cm under laparotomy in 2004. She has been disease free for 15 months. PMID- 16315907 TI - [Clinicopathological significance of pyrimidine nucleoside phosphorylase (PyNPase) and dihydropyrimidine dehydrogenase (DPD) in advanced colorectal cancer]. AB - We examined clinicopathological characteristics and prognoses of seventy advanced colorectal cancer cases by measuring pyrimidine nucleoside phosphorylase (PyNPase) and dihydropyrimidine dehydrogenase (DPD) in tumor and normal tissue. PyNPase activities in cancerous tissue obtained from resected were 82.7 +/- 41.9 U/mg protein, which were significantly higher than 37.2 +/- 24.0 U/mg protein in normal tissue (p < 0.001). On the other hand, DPD activities in cancerous tissue were significantly lower in normal tissue (p < 0.05). In cases with lymphnode metastases, PyNPase activities of cancerous tissue were significantly higher than that of no lymphnode metastases cases (p < 0.05). In cases with grade 2 side effects or higher by oral adjuvant chemotherapy, DPD activities in normal tissue were significantly lower than that of other cases (p < 0.05). With regard to Dukes' B and C cases that were resected curatively, PyNPase activities of cancerous tissue of higher group's prognosis were worse than that of the lower group. In the group received 5'-DFUR as adjuvant chemotherapy, non-recurrent survival rate of the group exhibiting higher PyNPase activities was better than that of the lower group. PMID- 16315908 TI - [Chemotherapy for patients with advanced/recurrent gastric cancer and the practice at our institute]. AB - Chemotherapy results for all 157 patients with advanced/recurrent gastric cancer at our institute were analyzed based on survival duration. The median survival time (MST) of all 157 patients was 426 days. The break down of the 157 patients are as follows: 59 patients with recurrence after curative operation, 47 patients with a radical grade C after non-curative operation and 51 patients with unresectable gastric cancer, and whose MST's were 590, 610 and 215 days, respectively. The standard chemotherapy for advanced/recurrent gastric cancer has not been established. Our data showed that chemotherapy for gastric cancer should be performed as a means of clinical study. If the patients had any exclusion criteria of clinical study or a proper clinical study did not exist, we would have recommended the TS-1 based regimen as the first-line chemotherapy, and the taxane based regimen as the second-line chemotherapy. PMID- 16315909 TI - [A study of recurrent pancreatic cancer with metastatic liver tumors after pancreatectomy]. AB - BACKGROUND: Pancreatectomy is the best treatment for pancreatic cancer. However, there is a high risk of post-operative complications, such as local recurrence, metastatic lymphadenopathy, carcinomatosa peritonitis and liver metastasis. Presently, there is no significant treatment that has yet had a strong impact on recurrent pancreatic cancer. This study was conducted concerning recurrent pancreatic cancer with metastatic liver tumors after pancreatectomy. SUBJECTS AND METHODS: Between April 1998 and March 2005, our institute treated recurrent pancreatic cancer patients with liver metastasis. This study consisted of groups who received therapy and those who did not, and their subsequent survival rate was also studied. RESULTS: Nine cases had a recurrent pancreatic cancer with metastatic liver tumors. According to the treatment modality, the study population was classified into two groups: one group consisted of 4 patients with no treatment and the other group consisted of 5 patients who received treatment with gemcitabine (GEM). The mean survival rate for the group who received no treatment was 6.6 months. The mean survival rate for the group who received the treatment was 22.3 months. In addition, one year survival rate for those who received no treatment was 0%. The one year survival rate for those who received the treatment was 75%. The group with treatment showed better results in overall mean survival rates, as well as a higher one-year survival rate than the group with no treatment. The GEM administration included both intraarterial infusion and intravenous infusion. CONCLUSIONS: Prolongation of the survival period was obtained by administration of GEM for recurrent pancreatic cancer with metastatic liver tumors and inoperable advanced pancreatic cancer. We attempted to utilize the active treatment for recurrent pancreatic cancer with metastatic liver tumors. Thus, it was indicated that the therapy can be effective against recurrent pancreatic cancer with metastatic liver tumors. PMID- 16315910 TI - [Indication and meaning of hepatic resection for metastatic liver tumors from gastric cancer]. AB - To clarify the efficacy and indication of hepatectomy in gastric cancer metastasis to the liver, we studied 26 liver metastases patients undergoing hepatectomy. The cumulative 5-year survival rations were 25.3%. A survival analysis showed that prognoses of patients n(-) or T2 group were statistically better than other group. Three patients underwent a repeat liver resection from gastric metastasis and survived for about thirty months. There was an example using chemotherapy which made a liver tumor removal operation possible to the patient who had multiple liver metastases. In conclusion, liver resection is a beneficial option and multimodal therapy including a repeat liver resection and systemic chemotherapy, may be important for improving the prognosis of patients with liver metastases from gastric cancer. PMID- 16315911 TI - [Effectiveness of intraperitoneal chemotherapy using new-aged drugs for the peritoneal dissemination of gastric cancer]. AB - We evaluated the effectiveness of intraperitoneal chemotherapy for peritoneal dissemination of gastric cancer. A total of 104 patients with primary gastric cancer with peritoneal dissemination were enrolled in this study. In 72 of the 104 patients with gastrectomy performed, 5 patients underwent CDDP-ip (50-100 mg/100-200 ml), 16 patients underwent CHPP (CDDP 300 mg, MMC 30 mg, ETP 150 mg/8 l/42-43 degrees C/60 min), and 17 patients underwent CMV-ip (CDDP 150 mg, MMC 20 mg, ETP 100 mg/1 l/60 min). The prognosis of patients who underwent CMV-ip was significantly better than those who received other therapies. In 26 patients with severe peritoneal dissemination who were treated with TS-1 (60 mg/m2) and taxane ip (docetaxel 40-60 mg/body or paclitaxel 120-180 mg/body), 9 of 18 responders performed complete cytoreduction. The median survival time (MST) in these 9 patients was 944 days and a 2-year survival rate was 63%. A multimodal therapy consisting of systemic chemotherapy, intraperitoneal chemotherapy and complete cytoreductive surgery may provide good prognosis to the gastric cancer patients with peritoneal dissemination. PMID- 16315912 TI - [Safety and efficacy of hypotonic CDDP intraperitoneal administration for gastric cancer with peritoneal dissemination]. AB - We examined safety and efficacy of hypotonic CDDP intraperitoneal administration followed by systemic chemotherapy using MTX/5-FU and UFT. Between 1998 and 2004, seven patients who had histologically proven gastric adenocarcinoma with peritoneal metastases underwent palliative gastrectomy at Niigata University Medical Hospital. For residual peritoneal tumors, 100 mg/body of CDDP diluted with distilled water was intraperitoneally administered to the patients before closure of abdominal wall and was drained 30 to 60 minutes after administration. During the postoperative period, a patient suffered from intraperitoneal abscess and another patient had a renal dysfunction with an increasing level of serum Cr (2.1 mg/dl). As adverse effects of the following systemic chemotherapy, three patients had grade 3 anemia and one had grade 3 leukopenia. The median time to progression was 109 days and the median survival time was 248 days. Although intraperitoneal CDDP administration is safe to be carried out intraoperatively, the effect on survival is not better than new anticancer drugs, such as TS-1 and paclitaxel. PMID- 16315913 TI - [The efficacy of intra-peritoneal chemotherapy for the patients with cytology positive gastric cancer]. AB - In this study, we assessed efficacy of repeated intra-peritoneal chemotherapy with CDDP for patients with cytology positive gastric cancer. The median survival time was 338 days, 1-year survival was 60% and 2-year survival was 45% of the patients. The POCY1 patients with repeated intra-peritoneal chemotherapy yielded a tendency to extend the survival rate than the patients without repeated intra peritoneal chemotherapy (p = 0.06). But, no differences were found between the survival rate of P1CY1 patients with or without repeated intra-peritoneal chemotherapy. The patients using more than 3 anti-cancer drugs yielded a tendency to have a better prognosis than the patients using 2 or less anti-cancer drugs (p = 0.09). There was a possibility to which the multidiscipline treatment was effective for the POCY1 gastric cancer patients. PMID- 16315914 TI - [Evaluation of peritoneal cytology in colorectal carcinoma]. AB - We investigated the incidence of free cancer cells in the peritoneal washings of 278 patients who had undergone surgery for colorectal cancer to evaluate its influence as a prognostic factor of the disease. Twenty-two cases (7.9%) were found to have malignant positive cytology (CY(+)). The rate of CY(+) in the cases with peritoneal dissemination (P(+)) was significantly higher than that in P(-) (66.7% vs 3.8%). In 244 cases, those who had tumors exposing to the peritoneum, both CY(+) and P(+) were observed highly in poorly differentiated adenocarcinoma. Among 18 P(+) cases, the rates of CY(+) were higher in both P3 and cur C than in P1, 2 and cur B. When restricted to 260 P(-) cases, CY(+) was observed more often in stage IV cases (14.3% vs 1.8%). Rate was significantly high in M+ (66.7%). Prognosis of 4 P(-) CY(+) cur A cases was as follows; 2 survived for a long time with no recurrence (20 and 60 months), 1 had curable liver metastases after half a year and obtained a 2 year disease free period after surgery, and another one died with brain, liver, and peritoneal recurrence one year later. The incidence of CY(+) is correlated with P(+); CY(+) increased when P(+) is extended more highly and incurable. CY(+) alone doesn't become a prognostic factor for peritoneal recurrences, because CY(+) is found rarely in curable P(-) cases. However, CY(+) is also associated with far advanced cancer with remote metastases, therefore we should consider the risk of such metastases for CY(+) cases with curable colorectal cancer. PMID- 16315915 TI - [Analysis of Reg IV expression in peritoneal dissemination of gastric cancer using real-time RT-PCR]. AB - We had performed a global analysis of the gene expression of gastric cancer cell lines established from malignant ascites to identify the novel markers for the detection of micro-metastasis in peritoneal cavity. One of the up-regulated genes is Reg IV, which is a member of the Reg gene family belonging to calcium dependent lectin (C-type lectin) gene superfamily. But the role of Reg IV in peritoneal dissemination is still unclear. We have examined the potential of Reg IV as a novel marker for the detection of peritoneal micro-metastases of gastric cancer. Reg IV expression was examined by quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Mean Reg IV mRNA expression levels in surgically resected specimens (n = 41) were more than 20-times higher than those in normal mucosa from those patients. Furthermore, Reg IV mRNA expression level in the peritoneal wash was strongly higher in peritoneal metastasis compared to those without peritoneal metastasis. These results suggest that Reg IV may be involved in peritoneal dissemination of gastric cancers and Reg IV would be a potential novel marker for peritoneal dissemination of gastric cancers. PMID- 16315916 TI - [A case of ascities decrease in malignant peritoneal mesothelioma by weekly intra peritoneal administration of cisplatin and paclitaxel]. AB - We report a case of malignant peritoneal mesothelioma (MPM) in a 63-year-old man. He had body weight loss and abdominal distension for one month, and was admitted to our hospital. Abdominal sonography showed a large mass occupying the right lower abdomen and an existence of a lot of ascites. Computed tomography and magnetic resonance image showed a lot of ascites and omentum cake. Cytology of the ascites was Class V but its histological classification was unknown. Then we performed biopsy of the tumor into the omentum with abdominal sonography. The histological diagnosis was MPM because the tumor cells showed positive for calretinin. He received a combination chemotherapy of weekly intra-peritoneal administration of cisplatin (70 mg/ day) and paclitaxel (100 mg/day). The ascites was decreased and per os (PO) was possible, but omentum cake was not changed. MPM was poor in prognosis and the control of ascites was difficult. We suggest that the chemotherapy of intra-peritoneal administration was a better procedure than others to control ascites with malignant tumors. PMID- 16315917 TI - [Intraperitoneal and intrapleural gemcitabine in patients with advanced pancreatic cancer]. AB - We performed intraperitoneal and intrapleural dosing gemcitabine (GEM) to eight patients with advanced pancreatic cancer having peritoneal or pleural carcinomatosis and evaluated its actions and safety. GEM (500 mg/m2) was infused into the abdominal cavity or thoracic cavity after drainage of peritoneal or pleural effusion. We checked the change of serum GEM concentration and the side effects after the GEM administration. Then, we repeated the GEM administration observing their systematic symptoms and evaluated the alteration of peritoneal or pleural effusion and cytology. Plasma concentration of GEM by infusing into the abdominal cavity or thoracic cavity was lower than by intravenous injection. In three of the five cases of peritoneal carcinomatosis, intraperitoneal administration revealed a decrease of peritoneal effusion. In two of the three cases of pleural carcinomatosis, intrapleural administration revealed a decrease of pleural effusion. Four cases had leukocytopenia of grade 1/2, three cases had thrombocytopenia, and two cases had alopecia as side effects, although all of them were minor side effects. Intraperitoneal and intrapleural dosing GEM had minor side effects and could improve QOL for the patients with advanced pancreatic cancer associated with peritoneal or pleural carcinomatosis. PMID- 16315918 TI - [Hepatic arterial infusion chemotherapy using W-spiral (WS) catheter]. AB - The authors investigated the usefulness of W-Spiral (WS) catheters for hepatic arterial infusion (HAI) chemotherapy especially from the viewpoint of a removable catheter after cessation of chemotherapy. A WS catheter was successfully inserted into the hepatic artery without a coil fixation in 40 out of 47 patients with hepatic malignant disease. A stable position of the catheter tip was maintained throughout the period in all cases. In 20 patients, the catheter was easily and uneventfully removed after chemotherapy, which resulted in improved patients' quality of life. An electron microscope examination revealed that there was no fibrin network formation on the surface of the catheter. Stenosis of the hepatic artery was confirmed in 7 out of 12 patients with 5 Fr catheter by 3DCT angiography, whereas patency of the artery was well preserved in 7 of 8 cases with 3.3 Fr catheter. This catheter has a special shape-memory alloy in its tip, which allows a preferable fixation without coils and removal of the catheter if desired. HAI using this catheter and subsequent removal of it is a reasonable strategy especially for patients who need limited courses of chemotherapy. PMID- 16315919 TI - [Five cases of locally advanced rectal cancer or local recurrence performed intra arterial infusion chemotherapy via the internal iliac artery]. AB - Intra-arterial infusion chemotherapy via the internal iliac artery was performed in 5 patients with locally advanced rectal cancer or recurrent rectal cancer. Arterial infusion chemotherapy was conducted into the internal iliac artery via bilateral femoral artery following a blood flow change with a coil. 5 FU 500 mg and l-leucovorin 125 mg/m2 were injected weekly. An average time of injections or its duration was 40 (17-74) times or 12.8 (5-23) months, respectively. Disappearance or improvement of symptoms was observed in 4 cases. A decrease of tumor size observed by CT was in 2 cases and a decrease of blood CEA level was in 3 cases. As for the complication of arterial infusion chemotherapy, dermatopathy was found in all of the cases, and sensory disturbance of lower extremities was in 3 cases, infection was in 2 cases and catheter obstruction was in 2 cases. A decrease of dosage or abundance of continuation was done during the course due to complications. Two patients with primary cancer died 1-3 years after the treatment, and 2 patients with recurrence died 7 months to 1 year after the treatment. One patient with primary cancer is continuing the treatment for the last 2 years though multiple metastatic diseases have been confirmed. PMID- 16315920 TI - [Chemotherapy for liver metastasis of colorectal cancer]. AB - In colorectal cancer, liver metastasis is the most common and most important prognostic factor. Although surgical resection is the first choice of treatment for liver metastasis of colorectal cancer, there are many cases we cannot choose the surgical treatment. The chemotherapy is very important in such cases. We examined 18 cases of unresectable liver metastases from colorectal cancer which were adapted a hepatic arterial infusion of 5-FU (HAI) with a weekly high-dose infusion method (WHF) as the first-line treatment, and then systemic chemotherapy of CPT-11 in combination with 5-FU as the second-line treatment. The response rate of this treatment is 72% (13/18) and the 1-, 2-, 3-year survival rates were 100% (16/16), 83% (10/12), and 50% (5/10), respectively. The combination chemotherapy of HAI with systemic chemotherapy using CPT-11 seemed to be an effective treatment method. PMID- 16315921 TI - [The docetaxel radiosensitization experience for the treatment of unresectable esophageal cancer]. AB - BACKGROUND: Docetaxel is an increasingly important drug for the treatment of esophageal cancer. The docetaxel radiosensitization has been established in cancer cell lines. The therapeutic response and toxicity of a weekly docetaxel in combination with radiotherapy for unresectable esophageal cancer were examined. METHODS: Ten patients with locally advanced or metastatic squamous cell esophageal cancer were recruited in the following protocol. The median age was 65.7 years. Patients received radiation in 2 Gy single daily fractions to a total dose of 60 Gy. Docetaxel (10 mg/m2) was administered weekly for 6 consecutive weeks. RESULTS: One patient could not be evaluated. The overall response rate was 77% with 11% CR and 66% PRs. Mild grade 2 leukocytes toxicity was observed in 2/10 patients, which enforced the treatment absence for 7-14 days. Grade 2 stomatitis was noted in one patient. No severe grade 3 adverse effects were observed. CONCLUSION: It is concluded that low dose docetaxel with radiotherapy is feasible and, a high response rate can be expected. Toxicity is modest, and this protocol may be useful for the outpatients or neoadjuvant chemotherapy. PMID- 16315922 TI - [The evaluation of chemoradiotherapy to unresectable hepatobiliary cancers]. AB - Today, there are few effective treatment options to unresectable hepatobiliary cancers. We enforced chemoradiotherapy to 7 patients who had far advanced hilar cholangiocarcinomas or hepatobiliary cancers because they could not receive radical operations. Then, we examined the effectiveness of this therapy. The radiation method was a conformation radiotherapy to 5 patients and the combination of conformation radiotherapy and RALS to 2 patients irradiated once with the radiation of 2-3 Gy. The menu of chemotherapy was the combination of 5 FU (1,250 2,500 mg/week) and CDDP (10-50 mg/week) by intravenous infusion or injection to the hepatic artery in all patients. By this therapy, 6 out of 7 patients were able to live for one year or more, and the median survival time was 1.41 years (0.65-2.65). Only two patients were judged as clinical PR by computed tomography, but the value of the tumor marker after radiotherapy was 85.7% (6/7), a decrease of 1/3 or less before the treatment. As for side effects, nausea, vomit, and appetite loss were observed in some patients. However, they were not so severe and the treatment could be easily restarted. Though various examinations are necessary, the chemoradiotherapy is expected to be useful for unresectable hepatobiliary cancers. PMID- 16315923 TI - [Gemcitabine concurrent with radiation for locally advanced pancreatic cancer]. AB - Gemcitabine (GEM) concurrent with radiation is clinically not well defined. We herein report four cases of chemo-radiotherapy against locally advanced pancreatic cancer using low-dose GEM concurrent with extra-beam radiation. A total of eight cases entered the study. Three were resected and five were non resected cases. Intraoperative radiation was carried out in every case using an 8 or 10 centimeter cone with a radiation dose of 25 Gy. Postoperative radiation was 2 Gy per day on weekdays for 5 weeks. Four cases were concurrent with low-dose GEM (40 mg/m2) twice a week, whereas the other four were radiation only. With the use of GEM concurrent with radiation, tumor markers decreased more than 80 percent regardless of the tumor resection. CT scan confirmed a necrotic change and the decrease of the tumor size. In conclusion, low dose GEM concurrent with radiation therapy may be a promising therapeutic choice for the local control of advanced pancreatic cancers. PMID- 16315924 TI - [Weekly docetaxel in combination with concomitant radiotherapy for esophageal cancer]. AB - Docetaxel (DOC) has demonstrated an activity as a radiation sensitizer in head and neck cancer. We have experienced four cases of esophageal cancer treated by weekly DOC in combination with concomitant radiotherapy. Three cases of advanced esophageal cancer and a case of relapsed esophageal cancer after administration of 5-FU and CDDP were enrolled in this pilot trial. The schedule of treatment included a weekly administration of DOC (10 mg/m2/w) and six weeks of radiotherapy in 2.0 Gy daily fractions up to 60 Gy. Combination treatments were completed successfully without any side effects except a case of developed grade 1 dysphagia. Two cases showed a complete response and others showed a partial response. One case showed a complete response developed abdominal para aortic lymph node and liver recurrence 12 months after the treatment. However, the targeted lesion within radiotherapy for this patient still showed a complete response. A weekly administration of low dose DOC in combination with radiotherapy was very effective for advanced esophageal cancer. DOC has demonstrated an activity as a radiation sensitizer. PMID- 16315925 TI - [A case report of complete obstruction of the left main bronchus caused by recurrent esophageal carcinoma with pulmonary embolism of right lung after surgery for esophageal cancer]. AB - A 54-year-old woman complained of dyspnea, due to complete obstruction of the left main bronchus caused by recurrent esophageal carcinoma, was transferred to our department about two months after curative resection (with preoperative chemotherapy for T4 cancer) in July 2004. She suddenly developed a severe shortness of breath with anxiety, and arterial blood gas analysis revealed a PaO2 of 25 mmHg (FiO2 1.0). The presence of pulmonary embolism was diagnosed by pulmonary perfusion scintigraphy. Thrombolytic therapy with urokinase was started to keep the air way. Fogarty catheter and bronchoscopic Nd-YAG laser treatment was performed. After that, an expandable metallic stent (EMS) was placed at the site of obstruction without any troubles, and there was a striking improvement in her condition of respiration and atelectasis. The radiation therapy was initiated and completed safely. The tumor lesion had disappeared on CT scan after the radiation therapy (a total dose of 50 Gy). We experienced a case that could be rescued from an advanced respiratory failure caused by one side air way obstruction and another side's blood circulation disorder. PMID- 16315926 TI - [A long-term survival case with multiple liver metastases from duodenal gastrointestinal stromal tumor that was drastically reduced by the treatment with imatinib]. AB - Constitutive activation of c-kit receptor tyrosine kinase is a critical factor in the pathogenesis of gastrointestinal stromal tumors. Imatinib mesylate (IM), a selective tyrosine kinase inhibitor, has been shown in clinical studies to work against such tumors. But there is little information on whether a combination of IM and surgical treatment can prolong survival in a case of unresectable multiple liver metastases. We report a case of postoperative recurrence of gastrointestinal stromal tumor (GIST) treated by the tyrosine kinase inhibitor IM and surgical treatment. Therefore, we discuss some important implications. This 37-year-old Japanese man underwent a pancreaticoduodenectomy for GIST of the duodenum in January 2003. The postoperative course was good at first, but 3 months after the initial operation, MRI showed multiple liver metastases. The patient was treated with 400 mg of IM once daily with only weak liver dysfunction as a side effect. The initial response to treatment of CR continued for 18 months. Currently, IM is the first-line therapy for non-resectable GISTs. As the mechanisms of recurrence and resistance to imatinib in GIST remain unclear, they should be intensively performed in the sight of both clinical and molecular biological viewpoints. Further examination in more cases of recurrent GIST is also necessary to estimate the effectiveness of treatment with IM. PMID- 16315927 TI - [The appreciation and limitation of regional gastrectomy for early gastric cancer]. AB - We performed limited surgical treatments, including modified radical gastrectomy (D1+ alpha) for 7 patients, segmental gastrectomy for 9 patients and regional gastrectomy for 13 patients from September 1999 to December 2004. Eligibility criteria were as follows: 1) mucosal gastric cancer without an ulcer and/or scar; 2) contraindication to EMR; 3) located at M or L area; 4) negative for lymph node metastasis in clinical and surgical findings. Regional gastrectomy, especially in the patients whose gastric cancer located at the side of greater curvature, improved postoperative remnant gastric function according to the average of half emptying times determined by RI scintigraphy, endoscopic examination and so on. However, the accuracy of mucosal cancer without lymph node metastasis in clinical and surgical findings was 69% and there was a patient with recurrence of lymph node after regional gastrectomy. These limited surgical treatments were thought to be limited due to more careful diagnosis as to sentinel lymph node navigation surgery in the future. PMID- 16315928 TI - [A case of intraperitoneal paclitaxel administration in a gastric cancer patient with severely impaired renal function]. AB - This study was designed to evaluate the pharmacokinetics and toxicity of paclitaxel, administered via an intraperitoneal route for a gastric cancer patient with renal dysfunction. The patient was a woman in her 50's, who had been diagnosed with severe renal dysfunction but no treatment history was known. She complained of dyspnea for a large quantity of ascites and was urgently hospitalized. It was diagnosed as gastric cancer with peritoneal dissemination. At this hospital, PTX was administered weekly intraperitoneally through an infusion port without complication. This result suggested that intraperitoneal PTX chemotherapy for a patient with renal dysfunction was a safe treatment. PMID- 16315929 TI - [A trial of TS-1 administration on the basis of the pharmacokinetic study for an advanced gastric cancer patient with impaired renal function]. AB - TS-1 is an antitumor drug including 5-chloro-2,4 dihydroxypyridine (CDHP), which inhibits dihydriopyrimidine dehydrogenase (DPD) activity selectively in metabolism of 5-FU. However, TS-1 therapy tends to increase adverse events for patients with impaired renal function due to excessively high blood concentration of 5-FU, because CDHP is mainly excreted into the urine. In a 67-year-old male with advanced gastric cancer, renal dysfunction occurred during TS-1 administration as its adverse event. We studied the pharmacokinetics of 5-FU, which were analyzed on the T1/2 value and the AUC (0-infinity) of 5-FU with a single and consecutive TS-1 administration, and estimated an optimal TS-1 administration regimen for this patient. The regimen is 60 mg/body/day given in one divided dose for 28 days consecutively followed by 14 days rest. This regimen enabled a continuation of TS-1 treatment for the patient. In conclusion, individual dose adjustment using pharmacokinetic study of 5-FU might be beneficial to patients with impaired renal function. PMID- 16315930 TI - [An effective weekly paclitaxel administration for gastric cancer with malignant ascites--a case report]. AB - A 71 year-old woman underwent total gastrectomy for advanced gastric cancer of p stage IV (pathological findings: por1 type 3 pT3, pN3 (12p: 1/1, 16b1 int: 3/3, 16b1 lat: 2/2), P1, CY1, H0) in March 2002. She was treated with the double modulation therapy of MTX/CDDP/5-FU intraperitoneally after the surgery. After leaving the hospital, she was carrying out the chemotherapy with MTX/5-FU continually. In August 2002, she became hospitalized once again because an appetite decrease and diarrhea appeared. CT of abdomen showed that malignant ascites had obviously accumulated, and she was admitted. Because it was conceivable in all cases of an inflammation by the chemical stimulation that originated in an anticancer drug, we suspended the intraperitoneal chemotherapy. Paclitaxel 90 mg/body administration was started intravenously on a weekly basis from the end of the same month. Those symptoms improved and she was discharged from the hospital, and was continued the paclitaxel administration. In CT of the abdomen that was taken in November in 2002, malignant ascites had obviously been decreasing and disappeared completely after that. PMID- 16315931 TI - [A case of advanced gastric cancer with liver and lung metastases markedly responded to TS-1-based sequential chemotherapy]. AB - A 61-year-old man with multiple liver and lung metastases from advanced gastric cancer was admitted to our hospital. We selected the patient from the outpatient department and started a single administration of TS-1 as a first line chemotherapy. TS-1 was markedly effective and the CEA level decreased 4,528 ng/ml to 44 ng/ml only in three months. The clinical response was assessed to be partial response (PR) comparable to complete response (CR) according to the tumor regression effect. The effect had been continued for almost six months. Because the CEA level elevated again, we estimated that the tumor acquired a drug tolerance to TS-1. Therefore, we applied CDDP with TS-1 as a second line chemotherapy. Unfortunately, it was not effective. Then we combined paclitaxel (PTX) to TS-1. The CEA level was remarkably reduced again, but transiently. Thereafter, we continued a sequential administration of TS-1 plus other drugs (CPT-11, docetaxel and MMC). Over 2 years, the patient is alive with a good quality of life. PMID- 16315932 TI - [A case of gastric cancer with liver metastasis resected after the combination chemotherapy with CDDP and TS-1]. AB - A 44 year-old woman presented with epigastralgia in March 2004 was diagnosed as having type II gastric cancer by gastrofiberscope, and histological diagnosis of biopsy specimens was group V (tub2, por1). Since multiple liver metastases were observed in subsegments 2, 3, and 5 by abdominal ultrasonography, systemic chemotherapy was first conducted for tumor down sizing. The patient was treated by three cycles of the one month regimen (CDDP 70 mg/m2, day 8, for 24hrs, and TS 1 80 mg/m2, day 1-14; 2 week cessation of the drugs). CT scan taken on May 16, 2005 revealed that the tumor diameters in subsegments 2, 3, and 5 were 1 cm, 3 cm, and 1.5 cm, respectively. On September 14, liver tumors were markedly shrunk. Tumor in subsegment 2 became undetectable, and the diameters in those in subsegments 3 and 5 were 1.5 cm, and 0.5 cm, respectively. On September 28, a distal gastrectomy associated with S3 partial hepatectomy and microwave coagulation therapy for S5 tumor was performed without any macroscopic residual lesions. The prognosis of liver metastasis of gastric cancer is generally poor, and there is no comprehensive therapy. The marked clinical response in this patient suggests that this combination therapy with CDDP and TS-1 might be a promising preoperative chemothepapy for unresectable gastric cancer. PMID- 16315933 TI - [Rectal stenosis due to Schnitzler metastasis following surgery for gastric cancer--a case successfully treated with TS-1 and CDDP combination chemotherapy]. AB - The patient, a 40-year-old woman, underwent total gastrectomy and excision of the pancreatic tail, spleen and gallbladder for gastric cancer in September 2000. The lesion was judged to be P1, SE, H0, N2 and Stage IV and the patient was managed on a regular schedule as an outpatient. In September 2004, she passed blood stained feces and rectal palpation detected a hard nodule at the anterior rectal wall. A fiber optic examination of the sigmoid colon detected an ulcerous lesion with a hemorrhage at the anterior rectal wall. A biopsy revealed the lesion to be Group V poorly differentiated adenocarcinoma. Starting in October 2004, 100 mg/day of TS-1 was administered for 3 weeks; intravenous drip infusion of 100 mg/body of CDDP was conducted in the second week for a period of 24 hours. After 3 courses of this regimen, a fiber optic examination of the colon conducted in February 2005 no longer detected the rectal tumor, leaving only a cicatrix. Upon a CT examination, the para-aortic lymph nodes that had been enlarged were notably reduced in size and an improvement was eminent in the hypertrophic rectal wall. The patient no longer experienced constipation or melena. Her clinical course is being observed while an oral administration of 100 mg/day of TS-1 continues. PMID- 16315934 TI - [A case of pseudo aneurysm of common hepatic artery related to hepatic arterial infusion chemotherapy]. AB - A 87-year-old male underwent the extended hepatic left lobectomy and the partial hepatectomy (S7, S8) for liver hepatic metastasis from the rectal cancer in March 2003. The reserver was implanted by the GDA-coil method from the right femoral artery, and WHF was enforced 10 times. Epigastric and back pain appeared from January 2004. The pain became strong gradually. MRI and CT scan were examined after hospitalization. These imaging tests showed the pseudo aneurysm of common hepatic artery. Therefore, an emergency angiography was performed, and pseudo aneurysm of the common hepatic artery was embolized with 36 metallic coils. After the embolization, the pain had disappeared suddenly. The patient was discharged the 7th day after embolization. Hepatic arterial infusion chemotherapy is considered safe with respect to blood and non blood toxicity, which was compared with systemic chemotherapy. However, there were also complications like this case, and caution is required. PMID- 16315935 TI - [A case of peritoneal dissemination disappeared by CPT-11 + TS-1 combination chemotherapy]. AB - A patient is a 35-year-old man. By a diagnosis of descending colon cancer, descending colon ablative operation and D1 lymph node dissection were performed on April 22, 2004. It was P3H0N1SE, Stage IV in perioperative findings. Abdominal CT showed peritoneal dissemination of 1.7 cm at the right under the abdominal wall wound and 1.2 cm in the rectovesical pouch on May 18, 2004. CPT-11 + TS-1 combination chemotherapy was started on June 22nd. In the five weeks of the combination chemotherapy, continuous infusion of CPT-11 (150 mg/body day 1 and 15) was twice administered, and oral administration of TS-1 (120 mg/body/day) was given for 3 weeks (day 1-21). Peritoneal dissemination disappeared after the two course end, and we judged it as CR. Furthermore, we were certain that we obtained CR after the three course end. The adverse event was only neutropenia of grade 1. The fourth course was not administered, but recurrence has not been observed. Abdominal CT showed no recurrence on March 3, 2005 since the combination chemotherapy ended 6 months ago. PMID- 16315936 TI - [A case of an increase in resectability with preoperative chemotherapy TS-1 combined with CPT-11 for unresectable rectal cancer in downstaging]. AB - A 50-year-old woman visited our hospital with a chief complaint of lower abdominal mass. The patient was diagnosed with rectal cancer using colonoscopy and also diagnosed with unresectable rectal cancer because abdominal CT revealed metastases to the liver, lung and lymph node located porta hepatis. The patient was treated with TS-1 combined with CPT-11. The TS-1 (80 mg/m2) was orally administered for 2 weeks and followed by a 2 week interval, and CPT-11 (80 mg/m2) was simultaneously administered biweekly. One cycle of chemotherapy was 28 days. The patient experienced grade 1 leukocytopenia and neutropenia. Abdominal CT revealed partial response after 2 cycles. After 6 cycles, the patient was subjected to curative operation. Pathological efficacy was Grade 1a at lymph node metastasis and Grade 3 at liver metastasis. TS-1 combined with CPT-11 regimen was very feasible and convenient, and obtained a good compliance. So this regimen was promising for unresectable colorectal cancer. In the future, this regimen will be verified in phase III clinical trial and compared with FOLFIRI and FOLFOX regimens. PMID- 16315937 TI - [A case of metastatic colon cancer treated with oxaliplatin combination chemotherapy]. AB - A 77-year-old male had been operated for ascending colon cancer with liver metastases. After hepatic artery injection therapy and CPT-11 plus 5'-DFUR combination therapy, oxaliplatin-based systemic chemotherapy was performed. Consequently, the tumor size was controlled for about 10 months. During the chemotherapy, we observed grade 3 neurological toxicity, but not grade 3-4 blood toxicity. Thus, the QOL of the patient was properly maintained. We conclude that the oxaliplatin-based chemotherapy may be useful for patients suffering 5-FU and CPT-11 resistant metastatic colorectal cancer in Japan. PMID- 16315938 TI - [A case of anal fistula cancer probably developing from intraluminal dissemination of rectal cancer]. AB - A 63-year-old man with a history of anal fistula was admitted to our hospital because the anal pain didn't disappear after the operation. On digital examination, a hard mass measuring 3.0 cm in diameter was found at the anal canal. Biopsy of the mass showed moderately differentiated adenocarcinoma. Colonoscopy revealed another rectal cancer at 15 cm from anal verge. Biopsy of the tumor also showed moderately differentiated adenocarcinoma, resembling the anal canal tumor. Because the histological findings of both tumors were nearly identical, we considered that cancer cells from the rectal cancer had been implanted and developed the metastatic tumor in the anal fistula. The patient underwent anterior resection for the rectal cancer, and a local resection for the anal canal cancer. Immunohistochemical staining for Ki-67, p53, Muc2, CD10, CK-7, and CK-20 revealed similar patterns in both tumors. Additionally, genetic analysis for p53, K-ras, and MSI revealed similar patterns in both tumors. We may suggest from these results that cancer cells from the rectal cancer had been implanted and developed the metastatic tumor in the anal canal. PMID- 16315939 TI - [A case of chemoradiation with tissue expander for local reccurence of rectal cancer]. AB - The patient was a 64-year-old female who underwent an abdomino-perineal resection for advanced lower rectal carcinoma. The tumor was a mucinous adenocarcinoma, type 2 in the Japanese classification of colorectal carcinoma, and was measured 4.5 cm in size. Histologically, the tumor was considered to be stage II (a2, n1, P0, H0). She received adjuvant chemotherapy with 5-FU/LV. After 7 months from the surgery, the patient developed a hip pain. Pelvic CT scan revealed a 4.5 x 6.0 cm mass anterior to the sacrum. Consequently, a diagnosis of local recurrence with sacrum invasion was made. An operation, involving tissue expander placement in the pelvis and pelvic floor was made by dexon mesh, was performed. Radiotherapy was given to the pelvic region at a total dose of 65 Gy (given 2.5 or 3 Gy each x 25 fragments). Chemotherapy with CPT-11 (100 mg/body) and 5'-DFUR (800 mg/body) was administered for 3 cycles. CEA (ng/ml) was decreased within a normal range from maximum 13.8. No progression was detected on CT, and Hip pain had disappeared. Abscess in the pelvis by tumor necrosis was managed conservatively. In this case, tumor marker is within a normal range over 2 years and QOL was improved, suggesting that chemoradiation with a tissue expander may be effective. PMID- 16315940 TI - [Two cases of postoperative local skin recurrence of breast cancer successfully treated with chemotherapy of irinotecan hydrochloride (CPT-11)]. AB - Breast cancer is a neoplasm which is considered well-responsive to chemotherapy, radiation therapy and endocrine therapy. However, recurrent breast cancer will grow developing a tolerance to the variety of therapies. We reported two cases of recurrent breast cancer which showed a good response to CPT-11. These tumors were already refractory to anthracycline and taxanes. A weekly CPT-11 infusion therapy was effective to the recurrent skin and soft tissue lesions (CR and PR), and the tumor markers (CEA and CA15-3) markedly declined. CPT-11 is a useful treatment for multiple drug resistant recurrent breast cancer. PMID- 16315941 TI - [A case of breast cancer detected by MRI mammography after Hollywood syndrome]. AB - A-64-year-old woman, who had been treated with augmentation mammaplasty 40 years ago, came to our hospital complaining of left breast pain. The mass was ill defined, located in the upper outer quadrant area of her breast, and was 2 cm in diameter. MRI examination showed that the tumor had a spiculation and an irregular edge. There were no regional lymph nodes in her axilla and supra subscapular. The diagnosis was Class IV by the fine needle aspiration biopsy cytology. We diagnosed the left breast cancer being in T2N0M0, Stage IIA, then we carried out Bt (Auchincloss method) and Sentinel lymph node biopsy (SLNB). There were metastatic cancer cells in the sentinel lymph node. So, we added level II lymph nodes dissection. The histological diagnosis was papillotubular carcinoma, f+, n+ (8/11). The endocrine receptor status of the tumor was ER+, PgR+ and the HER2/neu score was 0. There was paraffinoma in the non-cancer area. We dosed 6 cycles of FEC chemotherapy (CPA 800 mg, EPI 80 mg, 5-FU 750 mg/body x 1 cycle). We recognized no side effects of the chemotherapy for the patient. PMID- 16315942 TI - [Combination immunotherapy using autologous tumor-stimulated lymphocytes and trastuzumab (Herceptin) for the patients with recurrent breast cancer]. AB - PURPOSE: We report two patients with refractory recurrent breast cancer (HER2/neu: +) postoperatively, who had failed response to the available conventional chemotherapy of CAF (cyclophosphamide, adriamycin, 5-fluorouracil) and docetaxel, etc. They markedly responded to the combination immunotherapy using intraperitumoral injections of autologous tumor cell-stimulated T lymphocyte (AuTL) and trastuzumab (Herceptin), an anti-HER2 monoclonal antibody. METHODS: AuTLs were administrated directly into the recurrent tumor by intraperitumoral injections biweekly and trastuzumab was infused systemically every week. The treatments were repeated for 6 and 11 injections in the patients, respectively. The total administered T cells had reached to 3.8 x 10(9) and 6.4 x 10(9), respectively. The dosage of trastuzumab was 2 mg/kg in each patient. RESULTS: The carcinomatous pleural effusion had disappeared and was well controlled in patient 1 and a marked regression in injected fields in comparison to the size of the recurrent tumor before treatment was observed in patient 2. The tumor marker proteins (CEA, CA15-3, TPA) had also decreased significantly. The adverse effects of the immunotherapy were tolerable with grades 1-2 infusion reaction of fever, tachycardia and hypotension, but no cardiac dysfunction was observed. CONCLUSIONS: Clinical responses of recurrent breast cancer were observed in two patients after receiving the intra-peritumoral AuTL injection plus trastuzumab immunotherapy. These results showed that refractory recurrent breast cancer may be controlled effectively and safely by repeating the cellular immunotherapy combined with trastuzumab and suggested utility of combining these agents in clinical trial. PMID- 16315943 TI - [A case of advanced breast cancer with multiple lung and liver metastases successfully treated with multi-disciplinary therapy]. AB - We report a case of advanced breast cancer with multiple lung and liver metastases (T4bN1M1) achieving a significant improvement of QOL by multi disciplinary therapy. The patient was a 63-year-old woman with slight jaundice who had ascites and an ulcerative breast lump with multiple lung and liver metastases. A core needle biopsy for breast tumor led to a diagnosis of an invasive ductal carcinoma positive for HER2/neu protein expression. She received 6 cycles of tri-weekly docetaxel (60 mg/m2) and weekly trastuzumab. Although the ascites and the jaundice disappeared after chemotherapy, the response for breast tumor, metastatic sites in the lung and the liver were less satisfactory. Fifteen months later, she received radiation therapy so that metastasis in the brain was recognized. But she had no neurological symptoms. Multi-disciplinary therapy can improve patient's QOL and the clinical outcomes in Stage IV advanced breast cancer. PMID- 16315944 TI - [Two patients of recurrent breast cancer with carcinomatous pleurisy well controlled pleural effusion]. AB - We report two patients of recurrent breast cancer with carcinomatous pleurisy well controlled pleural effusion. One patient is a 49-year-old woman. She underwent radical mastectomy for right breast cancer in September 1993. She suffered from multiple liver metastases in June 2000, so CEF therapy contained hepatic arterial infusion chemotherapy and extended right lobectomy of the liver were performed in December 2001. Right pleural effusion was detected in December 2003, then, pleurodesis was carried out with OK-432 after thoracic drainage. After pleurodesis, a weekly paclitaxel therapy was started and she was taking the regimen continuously. Another patient is a 55-year-old woman. She underwent radical mastectomy for left breast cancer in September 1999. Local recurrent lesions on the left chest and left pleural effusion were found in May 2003. After thoracic drainage, infectious pleurisy was complicated, so the drainage tube was removed after the therapy for preventing infection. After pleurodesis, CE therapy followed by peroral chemo-endocrine therapy was performed. Both of the two patients are receiving outpatient treatment without recurrent pleural effusion as of July 2005. PMID- 16315945 TI - [A case of resectable solitary liver metastasis from breast cancer]. AB - We report a case of resectable solitary liver metastasis from breast cancer. A 53 year-old woman underwent a pectoral muscle-preserving mastectomy for T1 N0M0 (Stage I) left breast cancer in May 2000. Histopathologic diagnosis was medullary carcinoma. Thereafter, the patient was followed up with adjuvant chemotherapy. A solitary tumor in the right lobe of the liver at S6 was found 10 months after mastectomy by ultrasonography. With a diagnosis of liver metastasis from breast cancer, right hepatectomy was performed in May 2001. At present, she remains disease free for 4 years after hepatectomy. We think that a surgical procedure for liver metastasis from breast cancer, if possible, is beneficial in prolonged survival. PMID- 16315946 TI - [A case of successful treatment using "wrapping therapy" for huge hepatocellular carcinoma]. AB - This case is about a male in his 50's. In May 2004, he consulted a nearby doctor with abdominal pain and was pointed out a huge mass in the liver. He was diagnosed as hepatocellular carcinoma over 10 cm in diameter with chronic hepatitis type B. Because the tumor concurred with intra-hepatic metastasis along with lymph node metastasis, we planned transhepatic arterial embolization (TAE) as a first choice of treatment and performed twice. However, because of the tail inside part of the tumor was supplied from the gastroduodenal artery (GDA) and right gastroepiploic artery (RGEA) which were nutrient arteries, an effective TAE was impossible. The enhancement lesion that seemed viable was shown by enhanced CT. Therefore, a wrapping therapy (surgical decollateralization+silicon membrane sheeting) was performed under laparotomy for the purpose of cutting off blood supply to the part of the viable lesion in August of the same year. The tail inside part of the tumor that seemed viable fell into necrosis by wrapping alone, and the serum PIVKA-II level decreased within normal limit. As of ten months after the treatment, the patient is still alive without aggravation. PMID- 16315947 TI - [A long-term survival of the patient with hepatocellular carcinoma and advanced portal vein and bile duct tumor thrombosis successfully treated with multimodal treatments]. AB - We reported a 60-year-old male patient with hepatocellular carcinoma (HCC) of 5cm in diameter with advanced tumor thrombosis in the left main trunk of portal vein and bile duct. He was treated with multimodal treatments resulting in a long-term survival of more than 4 years. At first, he was treated with transcatheter arterial chemoembolization (TACE) in April 1999, but the therapeutic effect was insufficient. Therefore, we performed an extended left hepatic lobectomy in July. Since six HCCs appeared in a posterior segment in January 2000, we achieved microwave coagulation therapy under laparotomy. Because of diffuse relapse of HCCs in the same segment of the liver, we performed hepatic arterial chemotherapy (HAC) using low-dose CDDP and 5-FU. As a result, complete disappearance of the tumors was observed. A new lesion appeared in S7 in January 2001. We performed TACE, but relapsed in June, so we selected percutaneous radio-frequency ablation under CO2 angiography. Since a recurrent tumor was detected at the same therapeutic site with invasion to the diaphragm in September 2002, we performed a partial liver resection with synchronous excision of the diaphragm. We continued TACE and systemic chemotherapy for relapses in and out of the liver. Accordingly, he lived for over four years. We conclude that a long-term survival in this patient can be attributable to appropriate treatment selections and timing, such as hepatic resection, TACE, HAC and ablation therapies based on changes in diagnostic imaging and tumor markers. In addition, we have to pay attention to keep good hepatic reserve in order to continue treatment for recurrences of HCC. PMID- 16315948 TI - [Multiple hepatocellular carcinomas successfully treated by combined therapies]. AB - We report a case of advanced multiple hepatocellular carcinomas (HCC), successfully treated by combination therapies of transarterial chemoembolization (TAE), operation and radiofrequency ablation therapy (RFA). A 59 year-old man was detected with an increased serum alpha-fetoprotein (AFP) level and an ultrasonography (US) revealed three nodules of HCC at the right lobe in August 1998. He was successfully treated with a combination therapy of TAE and percutaneous ethanol injection therapy (PEIT) at the beginning. However, HCC had recurred quite often. TAE had been repeated for twelve times before right trisegmentectomy was performed in November 2002. Thirteen months after the operation, CT revealed a new lesion at subsegment 3, which was treated by RFA. Furthermore, a metastatic single lesion had appeared at the left lung, which was resected. He has still been well for 82 months since the first detection of HCC. PMID- 16315949 TI - [A case report of hepatocellular carcinoma (Vp4)--an attempt to reduce residual tumor thrombus using combination therapy (hepatic arterial infusion, hepatic arterial embolization and radiation)]. AB - A 57-year-old man was found to have elevated levels of HCC markers during an observation of chronic hepatitis C. Diffused hepatoma was involved in the posterior lobe, and tumor thrombus extended into the main portal vein (Vp4). Posterior segmentectomy and tumor thrombectomy were performed. But, CT scan 45 days after the operation showed an enhancement at the residual tumor thrombus in the posterior branch. The patient received a hepatic arterial infusion of 5-FU, followed by hepatic arterial embolization. Then, we chose radiation therapy to the tumor thrombus. The most recent CT showed no enhancement at the reduced tumor thrombus. There have been almost no reports of treatment for residual portal thrombus. Careful observations are necessary in such patients. PMID- 16315950 TI - [Sustained complete remission with reductive surgery plus percutaneous isolated hepatic perfusion (PIHP) for bilobar multiple hepatocellular carcinoma with portal venous tumor thrombus--a case report]. AB - For patients with multiple bilobar hepatocellular carcinoma (m-HCC) and/or advanced portal venous tumor thrombus (Vp3, 4), there has been no effective therapy, and the survival of more than 6 months was exceptional. Under these circumstances, we have developed a dual treatment (dual Tx) that combines reductive hepatectomy with percutaneous isolated hepatic perfusion (PIHP) for such patients. This dual Tx offers the high-rate of mid- and long-term survival in a subset of patients who had previously a dismal prognosis. Herein, we report a patient with Vp4 m-HCC who was successfully treated with dual Tx and survived for more than 2 years with a complete remission of hepatic tumors. A 53-year-old man had main tumors in the right lobe liver and multiple bilobar intrahepatic metastases (IM) with portal venous tumor thrombus reaching the portal trunk. He underwent an extended right hepatectomy with portal venous tumor thrombectomy, and subsequently PIHP twice in a 3-month period after reductive hepatectomy. After dual Tx, he had sustained complete remission for more than 2 years. He died because of obstruction of the superior vena cava by recurrent tumors in the mediastinum. His clinical course after treatment strongly indicates that the dual Tx should become a major treatment option for patients with Vp3, 4 m-HCC. PMID- 16315951 TI - [A case of multiple hepatocellular carcinoma with rapidly progressing bilateral portal vein tumor thrombosis--a complete remission achieved with dual treatment of reductive surgery plus percutaneous isolated hepatic perfusion]. AB - We herein report a case of multiple advanced hepatocellular carcinoma (HCC) with rapidly progressing portal vein tumor thrombosis (PVTT). All of the hepatic tumors have completely disappeared for more than two years by a dual treatment with reductive surgery plus percutaneous isolated hepatic perfusion (PIHP). A 55 year-old man was referred to our institution on June 30, 2003. The abdominal CT scan demonstrated multiple massive HCC in the entire liver with PVTT reaching the portal trunk (Vp4). Two weeks later, the PVTT rapidly progressed to the umbilical portion of the left portal vein, and to the confluence of the superior mesenteric vein and to the splenic vein. Thus, we semi electively performed an extended right hepatectomy together with thrombectomy of the PVTT. Subsequently, he underwent a repeated PIHP (1st; doxorubicin 90 mg/m2, 2nd doxorubicin 65 mg/m2). This treatment produced complete tumor clearance of all of the residual tumors in the left liver. In March 2005, he underwent partial pneumonectomy for a metastatic lung. This again resulted in normalization of serum AFP and PIVKA-II levels. Dual treatment is considered to be the strongest therapeutic modality for multiple advanced HCC with severe PVTT. In addition, a close follow-up is required because in such far advanced cases, metastatic lesions most likely recur in the liver but also in the distant organs. PMID- 16315952 TI - [A successful case of more than 5 years of disease free survival in hepatocellular carcinoma with tumor thrombus to the right main branch of portal vein after repeated hepatic resection following transhepatic arterial embolization (TAE)]. AB - A case is a male in his 50's. He received hepatic resections twice for hepatocellular carcinoma. Recurrence was pointed out in the residual liver with tumor thrombus to the right branch of the portal vein. The serum level of AFP was 648 ng/ml. We performed transhepatic arterial embolization (TAE) with Epi-ADM, CDDP, Lipiodol and spongel through the right hepatic artery before re hepatectomy. Posterior segmentectomy with an extraction of portal vein thrombus was performed. Pathological findings showed complete necrosis not only in the main tumor but in the portal vein thrombus also. He is alive for more than 5 years without recurrence after surgeries following pre-operative TAE. PMID- 16315953 TI - [A case of HCC with inferior caval vein tumor thrombus and multiple pulmonary metastases that remarkably responded to combination therapy of TS-1 and interferon-alpha]. AB - A 56-year-old male was admitted to our hospital for hepatoma with portal vein thrombus and multiple intrahepatic metastases. He underwent an extended left lobectomy and a partial resection of the liver in May 2002. After two weeks from the surgery, he received intra arterial 5-FU infusion chemotherapy combined with subcutaneous interferon-alpha injection to treat the lesions in the residual liver. Four months after the surgery, hepatic vein tumor thrombus appeared in the remnant liver and it extended to the inferior caval vein. And another 4 months later, multiple pulmonary metastases were detected with computed tomography and they grew rapidly in the view of their sizes and numbers. Because the combined therapy of 5-FU/interferon-alpha was not effective to distant metastases, we started a new regimen of oral administration of TS-1 and a subcutaneous interferon-alpha injection. After 1 treatment cool, hepatic vein thrombus was markedly reduced the size and vascularity in the CT. Multiple pulmonary metastases also decreased in their sizes and numbers. No adverse effect was seen during this treatment. It was suggested that a combination therapy of TS-1 and interferon-alpha may be one of the most effective treatment modalities against advanced HCC with distant metastasis. PMID- 16315954 TI - [A case report of recurrence of liver metastases from colorectal cancer, which seemed to have vanished for a time by intra-aortic chemotherapy]. AB - A 66-year-old man underwent lower anterior resection for rectum cancer with multiple liver metastases. The diagnosis was Stage IV well-differentiated tubular adeno carcinoma. n1H3M(-). As adjuvant chemotherapy, we chose intra-arterial infusion of 5-FU 1,500 mg/body/week from July 2002 to February 2003. The metastatic focuses have become smaller and smaller until they seemed to be scars. In July 2003, there appeared to be a liver metastasis again, which emerged at the same place of the first metastatic focus. We tried intra arterial infusion of DXR in addition to intra-venous infusion of CPT-11, but it seemed to have no effect. Finally, we performed a partial resection of the liver in March 2005. There were many metastatic legions that emerged, but we successfully carried through partial resections for all legions. The uniqueness of this case was the liver metastasis that had appeared at the same place of the first metastatic focus. As the strategy for liver metastasis of colon cancer, it is better to perform a surgical resection as soon as the focus becomes resectable. PMID- 16315955 TI - [Hepatic arterial infusion (HAI) chemotherapy achieved a complete response (CR) for multiple liver metastases of colorectal cancer--two case reports]. AB - We encountered two cases of concurrent multiple liver metastases of colorectal cancer in two patients who achieved a complete response (CR) to hepatic arterial infusion (HAI) chemotherapy. The first case is a 64-year old man who was found to have anemia, and a diagnosis of ascending colon cancer was made. There were 6 metastatic lesions in both lobes of the liver. A right hemicolectomy was performed. Postoperative chemotherapy consisted of 2 g of oral UFT-E as well as 3 mg of CDDP and 500 mg of 5-FU administered by HAI. The metastatic lesions disappeared after 9 courses of chemotherapy were carried out. A partial hepatectomy was performed in the scarred area. Histopathological examination revealed no cancer cells. No recurrence has been observed for 4 years and 10 months since achieving CR. The second case is a 69-year-old man who had thin stool and bloody stool. Rectal cancer was diagnosed. Five metastatic lesions were noted in both lobes of the liver. A low anterior resection was performed. l-LV 25 mg and 5-FU 500 mg were given postoperatively by HAI. After three courses of chemotherapy postoperatively, metastatic lesions disappeared. No recurrence has been noted for 2 years and 1 month since achieving CR. HAI chemotherapy is considered one of the useful treatment options for patients with multiple liver metastases of colorectal cancer. PMID- 16315956 TI - [A successful surgical treatment for solitary pulmonary and adrenal metastases after hepatic resection to hepatocellular carcinoma--a case report]. AB - We report a 65-year-old man who received a successful surgical treatment for both pulmonary and adrenal metastases after curative resection to hepatocellular carcinoma (HCC). He received a partial hepatic resection for HCC of the right hepatic lobe. Thirty-eight months after the first hepatic resection, a metastatic lesion of the right pulmonary lobe was detected by computed tomography (CT). He was orally administered of UFT (600 mg/day). After 6 months of the chemotherapy, a metastatic lesion of lung became decreased in size. However, a metastatic lesion of the right adrenal gland was detected by abdominal CT scan. Fifty six months after the first operation, we performed right adrenalectomy. A further 4 months later, we performed partial resection of the right pulmonary lobe. Eight months after the pulmonary resection, intrahepatic recurrence was detected and he received transcatheter arterial embolization (TAE) twice. Eighty one months after the first operation, he died of liver failure due to tumor progression. Surgical resection for metastases from HCC resulted in long-term survival even if there were extrahepatic metastases in two different sites. PMID- 16315957 TI - [Hand-assisted laparoscopic adrenalectomy to a solitary adrenal metastasis from lung cancer]. AB - A 75-year-old man with right chest pain was diagnosed with primary lung cancer in the right apical portion, and was treated with chemoradiotherapy because of a synchronous left adrenal tumor of 1.6 cm. Since the adrenal tumor did not increase in size for three months and there were no other relapses, the right upper lobectomy of the lung with the excision of the chest wall was performed. Afterward, an enlargement of the left adrenal tumor was encountered; he was admitted to our hospital for an operation. For the metastatic adrenal tumor from lung cancer, we performed a hand-assisted laparoscopic adrenalectomy. He recovered rapidly and returned to the previous hospital in two weeks after the operation. After the first report in 1992, the laparoscopic adrenalectomy has been established as the curative operation to adrenal benign tumor. The indication is being expanded to the malignancy because of the improvement of operation techniques and advancement of the operation equipments. We conclude that the laparoscopic adrenalectomy for malignant tumor is a safe, curative, and clinically useful surgical technique. PMID- 16315958 TI - [Successful treatment of combined intraarterial (5-fluorouracil and adriamycin and cisplatin) infusion chemotherapy for advanced hepatocellular carcinoma with multiple intrahepatic metastases and/or portal vein thrombosis--two case reports]. AB - Hepatic arterial infusion chemotherapy has been often selected as a therapeutic option for advanced hepatocellular carcinoma with multiple intrahepatic metastases and/or portal vein thrombosis. We successfully treated and obtained CR in the 2 cases of far advanced hepatocellular carcinoma with intraarterial infusion chemotherapy (FAP). Case 1 was a 71-year-old male who had advanced hepatocellular carcinoma with intrahepatic metastasis (IM3) which was recurrent after two surgeries. He received hepatic arterial infusion chemotherapy (FAP: 5 fluorouracil 500 mg/day: continuous infusion, day 1-5, adriamycin 10 mg/day, day 1, CDDP 10 mg/day, day 1). After 10 courses, abdominal CT revealed that the viable lesions had completely disappeared (CR). This patient is still alive with no recurrence after 21 months from the beginning of this treatment. Case 2 was a 74-year-old male who had advanced hepatocellular carcinoma with portal vein thrombi (Vp4) and intrahepatic metastasis (IM3). He received FAP arterial infusion chemotherapy with the same protocol as case 1. After 8 courses of this therapy, CT revealed that these lesions had disappeared (CR). This patient is still alive with no recurrence after 9 months from the beginning of this treatment. For 15 patients with advanced hepatocellular carcinoma using a same protocol, the response rate of this therapy was 33.3% (CR & PR). These findings suggested that combined arterial infusion chemotherapy of FAP may be feasible and a promising modality for the advanced HCC with intrahepatic metastases and/or portal vein thrombosis. PMID- 16315959 TI - [The hepatic artery injection chemotherapy and prostaglandin E1 administration for hepatocellular carcinoma invading the biliary tract with jaundice]. AB - A 69-year-old man had radiofrequency ablation therapy (following RFA) for type C cirrhosis with hepatoma (following HCC) of S7 in November 2001. Afterward the patient was followed as an outpatient, but he had been admitted to our hospital due to jaundice confirmed in March 2004. His abdominal wall appeared to be soft and flat, and we could not detect a tumor mass by palpating either. Even though he exhibited no actual symptom of anemia, jaundice was found in the bulbar conjunctiva at the time of admission. Laboratory findings showed a mild inflammation and anemia on his admission, and biochemical data showed a rise of hepatobiliary enzyme with jaundice. A rise of tumor marker (AFP, PIVKA-II) was recognized, too. We performed percutaneous transhepatic bile duct drainage (following PTBD) to decrease jaundice because abdominal echography and CT showed an extension of tumor thrombosis in bile duct and right hepatic duct by HCC of S8. However, a check of T-Bil. was 7.29 mg/dl and showed some slight decrease. Therefore, we administered prostaglandin E1 (following PGE1) at first with an intra-arterial injection catheter aiming to protect the hepatocyte. One week later, we performed hepatic artery injection chemotherapy (CDDP+5-FU) for four weeks. We confirmed a manifested improvement in T-Bil to be 1.92 mg/dl at the end of hepatic artery injections as well as a manifested decrease in hepatobiliary enzyme. We confirmed a decrease of HCC of S8 by abdominal CT, and the response rate was PR. Afterward the patient was conservatively treated even though pancytopenia was present, and was discharged from the hospital in June 2004. The hepatic artery injection chemotherapy used together with PGE1 was effective for the HCC patient with jaundice. PMID- 16315960 TI - [Efficacy of the combination therapy in early stage of recurrent cholangio celluler carcinoma (CCC)]. AB - We report a case of cholangio celluler carcinoma (CCC) with a good quality of life, in spite of the recurrence of peritoneum and portal hepatic lymph nodes (PHLN), due to the combination therapy that consisted of hepatic arterial infusion, systemic chemotherapy, radiation therapy and an insertion of a metallic stent into the bile duct. The patient was a 61-year-old man. Left hepatectomy was done due to multiple CCC. For the purpose of preventing the recurrence of CCC in residual liver, we performed an arterial infusion therapy. Ten months later, metastases to peritoneum and portal hepatic lymph nodes were found, so the resection of the peritoneum was performed, and cells in ascites were defined to be positive with cytology. After 2 years from the first operation, the size of portal hepatic lymph nodes had grown and both the billilbin and tumor marker levels had increased, so we started systemic chemotherapy, radiation therapy and insertion of a metallic stent into the bile duct. The tumor marker level decreased in a short time. Consequently, we inserted a metallic stent into the bile duct and radiation therapy was performed. Until the patient's death due to peritonitis carcinomatosa, the recurrence in residual liver occurred only once in three years after the first operation, and portal hepatic lymph nodes did not grow for two years after the recurrence. PMID- 16315961 TI - [A case of unresectable cholangiocellular carcinoma treated with surgery followed by combination chemotherapy]. AB - The patient was a 44-year-old man, who was investigated for lateral abdominal pain and liver dysfunction, and subsequently referred to our department with a diagnosis of unresectable intrahepatic cholangiocellular carcinoma (CCC). Radiological examinations revealed the huge mass in the right lobe of the liver with intrahepatic metastasis in the left lobe. The main tumor was surgically removed, but the metastases were not removed. A month after the operation, a subcutaneous implant reservoir was indwelled for repeated transcatheter hepatic arterial chemo infusion therapy (5-fluorouracil 500 mg/day continuous infusion, day 1-5, and CDDP 10 mg/day, day 1) from the right femoral artery. After 15 courses of home anti-cancer chemotherapy, abdominal CT revealed that the size of intrahepatic metastasis in the left lobe of the liver had not shown growth, whereas other metastitic sites popped up in the caudate lobe, which were free of chemical agent flow. There was no major complication related to the chemotherapy throughout the post-treatment course. Although he maintained a good level of QOL, he refused further chemotherapy due to depression. He died of liver failure 7 months after the operation. In conclusion, volume reduction surgery followed by transcatheter hepatic arterial chemo infusion might be promising as an effective therapy for non resectable CCC. PMID- 16315962 TI - [Two cases of successful local control with intermittent hepatic arterial infusion therapy using 5-FU and external radiation therapy for unresectable advanced gall bladder cancer]. AB - Unresectable advanced gall bladder cancer has a very poor prognosis. No good chemotherapeutic regimen is available. We report two cases of unresectable advanced gall bladder cancer patients who received the combination of intermittent hepatic arterial infusion chemotherapy and external radiation therapy. A 45-year-old male was alive for 576 days, and a 69-year-old male was alive for 246 days after the first diagnosis. They have been able to maintain a good quality of life without any severe adverse effects. PMID- 16315963 TI - [A case of advanced pancreatic cancer treated with chemoradiation]. AB - A 56-year-old man is presented with diarrhea, which he had experienced since February 2004. He was diagnosed as having advanced pancreatic cancer by enhanced abdominal CT scan in May 2004. He was diagnosed with unresectable pancreatic cancer, and treated with a combination of radiation (3 Gy/day) and injections of gemcitabine (GEM) 1,200 mg/week (800 mg/m2, BSA 1.6). Abdominal CT scan revealed a minor response (tumor diameter 5.7 x 4.8 --> 5.2 x 4.4). Accordingly, the improvement of performance status and reduction in serum levels of arcinoembryonic/carbohydrate antigen 19-9 (CA19-9) were observed. In July 2004, chemotherapy and radiotherapy were switched to GEM+UFT (UFT 360 mg/day, a total of 4,320 mg, GEM 1,200 mg according to the body mass, a total of 2,400 mg). The patient's performance state was stable for 6 months but serum levels of CA19-9 increased from March 2005, and he complained of diarrhea and back-pain. Therefore, the combination chemotherapy with GEM and cisplatin (CDDP) was started in April 2005, but there was no clinical effect. GEM and TS-1 are currently being administered. Pancreatic cancer is one of the worst prognoses of any malignant disease. Although the prognosis of unresectable pancreatic cancer is very poor, we presented a case where performance status and survival benefits were obtained by undergoing chemoradiation with GEM and combination chemotherapy with UFT and GEM. PMID- 16315964 TI - [A case of successful resection for recurrent intraductal papillary mucinous adenocarcinoma]. AB - Intraductal papillary-mucinous neoplasms (IPMN) of the pancreas have recently been defined and classified by the World Health Organization. In addition, many cases of IPMN continue to be recognized frequently. We report a case of a 65-year old female who underwent surgical resection of the pancreas twice within a period of 6 months for primary and recurrent IPMN. We first performed distal pancreatectomy with splenectomy for IPMN in the pacreatic body. A histopathological study revealed invasive adenocarcinoma and the negative margin of the pancreatic duct. A cystic mass in the uncinate process of the gland was identified by CT scan 6 months after the initial surgery. Complete pancreaticoduodenectomy was performed. Recurrent IPMN consisted of adenosquamous cell carcinoma. She currently is alive without evidence of disease at 3 years from the second resection. Recurrent disease in the residual pancreas suggests that a long-term surveillance is critical. PMID- 16315965 TI - [Portal vein stenting for portal hypertension caused by local recurrence following hepato-pancreatoduodenectomy for bile duct cancer]. AB - The patient was a 73-year-old man diagnosed with bile duct cancer. He underwent hepato-pancreatoduodenectomy and a combined resection of portal vein in July 2002. The post operative course was uneventful. In December 2003 he had a loss of consciousness, and was admitted to our hospital as an emergency patient. Abdominal CT scan and angiography revealed the presence of portal vein stenosis due to local recurrence of bile duct cancer resulting in portal hypertension. Thus, a metallic stent was placed in the portal vein to maintain portal blood flow. A non-covered metallic stent, 10 mm in diameter and 80 mm in length (SMART stent, Cordis Endovascular), was deployed through the stenotic portal vein. Portography after the stent placement showed a relief of the portal vein stenosis. Intravenous heparin administration was performed at a dose of 5,000 IU per day for 5 days after the stent placement, and this was followed by oral warfarin potassium administration. The clinical course was uneventful and the patient was discharged 21 days after portal stent placement. Portal stenting successfully improved portal hypertension due to stenosis of the portal vein without any complication. Our experience suggests that the portal vein stenting using the SMART stent is useful palliative therapy for portal vein stenosis. PMID- 16315966 TI - [A patient with middle bile duct cancer who showed rapid obstruction on contrast radiographs 1 week after insertion of a non-covered stent, but this case obtained a long-term stent opening and long survival after re-insertion of a covered stent]. AB - A 73-year-old female with jaundice was referred to our department and admitted. Blood examination on admission showed an abnormal liver function and an increase in CA19-9. Ultrasonography of the abdomen revealed a well-delineated round mass in the middle bile duct. After admission, PTBD was performed, and a complete obstruction was observed in the middle bile duct. Cytological examination of the bile demonstrated adenocarcinoma. Based on these findings, a diagnosis of middle bile duct cancer was made. Because of delirium and frequent wandering during the night due to dementia, a radical operation was abandoned, and a non-covered stent was placed. However, since 1 week after insertion of a non-covered stent, contrast radiography showed tumor protrusion (rapid obstruction) in the stent, and a covered stent was placed in the stent. The stent was open for 486 days after stent re-insertion. PTBD was performed for re-obstruction, but she died 596 days after admission. In patients for whom surgery is impossible for reasons other than tumor factors, internal fistula construction using a metallic stent, which neither impairs QOL nor shorten survival, can be indicated. PMID- 16315967 TI - [A case report of esophageal cancer with tracheoesophageal fistula--improving the quality of life by using a covered self-expandable metallic stent]. AB - We present the case of a 64-year-old male who was diagnosed with esophageal cancer with tracheal invasion and distant lymph node metastases, and he received chemoradiation therapy. The therapy resulted in complete remission. However, he was unable to eat anything because of missed swallowing caused by a large tracheoesophageal fistula. The placement of a covered self-expandable metallic stent (SEMS) improved his quality of life and palliated dysphagia for 3 months. Stenting in the cervical or upper esophagus may cause discomfort. However, the placement of a covered SEMS is one of the useful palliative treatments for esophageal cancer with tracheoesophageal fistula. PMID- 16315968 TI - [A successful treatment of percutaneous radio frequency ablation for advanced thyroid cancer]. AB - The patient was a 69-year-old woman. She received chemotherapy and radiation for thyroid tumor (undifferentiated cancer) following an operation in 1998. The chemotherapy was regularly repeated for relapse. The tumor increased gradually and came to cause dysphagia. She initially rejected nutritional management that depends on intravenous hyper alimentation or gastrostomy. After receiving an informed consent, we performed percutaneous radio frequency ablation (RFA) for the tumor using a Cool-tip needle on April 2, 2003. The algorithm of RFA was 9 min: 30 --> 120 W, 12 min: 50 --> 110 W, 9 min: 50 --> 100 W. After 2 days, the covered stent was implanted in the esophagus and an oral intake was started. Although the operation for undifferentiated thyroid cancer is controversial, we performed RFA and esophageal stenting for improving of QOL. This is the first case report in Japan. PMID- 16315969 TI - Quantitative motor unit action potentials (QMUAP) in whiplash patients with neck and upper-limb pain. AB - INTRODUCTION: Needle EMG studies in patients with whiplash symptoms to document presence of neural injury, such as cervical radiculopathy, are not available. OBJECTIVE: To document presence of motor unit action potential (MUAP) parameter changes compatible with neurogenic involvement in symptomatic-limb muscles in whiplash induced acute and chronic pain states. DESIGN: Retrospective review. SETTING: Out patient quantitative electromyography (QEMG) laboratory. PARTICIPANTS: 72 consecutive patients (mean age 43.6 years) who underwent QEMG between 1/2001 - 12/2004 for electrodiagnosis of neck and upper-limb pain related to auto-accidents (mean symptom duration 15.3 months). INTERVENTIONS: QEMG with MUAP parameter analysis. MAIN OUTCOME MEASURES: Amplitude, duration, size-index (SI), percentage of polyphasic units and firing rate analysis in chosen bilateral muscles representing C3-C8 myotomes. RESULTS: No spontaneous activity noted in muscles examined. Without subsetting symptom duration, symptomatic-side: asymptomatic-side MUAP parameter comparison showed significant increase in MUAP frequency on the symptomatic-side C6-muscle (10.4 +/- 1.3 vs. 9.7 +/- 1.4, p = < 0.05), and C7 muscle (10.0 +/- 1.2 vs. 9.1 +/- 1.1, p < or = 0.01). The symptomatic-side C6-muscle also showed increased percentage of polyphasic-MUAPs (23.7 +/- 13.1 vs. 15.9 +/- 12.7, p < 0.001). Patients with acute symptom duration (< 6 months, mean duration 3.4 +/- 1.4 months, N=26), showed increased percentage of polyphasic MUAPs in the symptomatic-side C6-muscle (27.0 +/- 12.3% vs. asymptomatic 22.2 +/- 17.6%, p < or = 0.05). In patients with chronic symptom duration (> or =6 months, mean duration 22.0 +/- 17.9 months, N=46), symptomatic side C6-muscle showed increased polyphasic MUAPs (22.2 +/- 13.2%) vs. asymptomatic (15.5 +/- 11.6%), p < or = 0.02, and higher firing rates (10.3 +/- 1.7 vs. 9.6 +/- 1.3 respectively, p < or = 0.003). CONCLUSIONS: QEMG changes suggest neural injury in symptomatic side C6 and C7 innervated muscles, even in the absence of spontaneous activity. In acute and chronic pain patients a higher percentage of polyphasic MUAPs is noted in the symptomatic side C6 muscle. In chronic pain patients higher MUAP frequencies are noted in the symptomatic side C6 muscle. PMID- 16315970 TI - Chaotic analysis of electromyography signal at low back and lower limb muscles during forward bending posture. AB - The present study aims to perform further investigation on muscular activity during forward bending posture by applying a nonlinear dynamic (chaotic) analysis method. The objective is to determine the characteristics of the low back and lower limb muscle electromyography (EMG) signal under chaotic analysis while maintaining a certain posture. Twelve subjects were asked to maintain postures of six stages in bending angles from 0 to 180 degrees, and the EMG signals of erector spinae (ES) at L1 and L5 levels, hamstrings, and gastrocnemius were recorded. Two important concepts to characterize deterministic chaos, Correlation Dimension (D2) and Lyapunov Exponents (lambda, LE), were applied to observe the chaotic characteristic of the EMG signals, and the results were also compared to the FFT based total power value. The EMG signals in all observed muscles during bending posture showed results of positive LE and high D2 at 5.5 to 7.5, which led us to classify EMG as a high dimension chaotic signal. The result obtained showed that the correlation dimension could be used as a reliable method to compare the EMG signal in various postures (or muscle contraction conditions). However, Lyapunov exponents did not show a significant difference of comparison result thus leading to the conclusion that LE could not be a reliable measure for high dimension chaotic system, such as an EMG signal. It was also shown that in both light and deep bending, the EMG signal of the low back muscles was of the same complexity level due to the D2 result. It was evident that somehow the low back muscle remained loaded in all bending stages which was contrary to the hypothesis that the low back muscle was less active during the deep bending, as was the case in most of the previous studies. The reason of such phenomenon was elucidated with use of the theory of muscular functional differentiation, including corticalization and spinalization. PMID- 16315971 TI - Peripheric and automatic neuropathy in children with type 1 diabetes mellitus: the effect of L-carnitine treatment on the peripheral and autonomic nervous system. AB - OBJECTIVES: To determine neuropathy frequency with electromyography (EMG) in asymptomatic diabetic children, and to demonstrate whether the electromyographical abnormalities noted improve after L-carnitine treatment. PATIENTS AND METHODS: This study was carried out on 51 type 1 diabetes mellitus patients (of whom, 26 were female; average age 12) and 21 healthy children as the control group. Thirty four patients, whose nerve conduction velocity (NCS) was diagnosed as pathological, were treated with L-carnitine (dosage: 2 g/m2/day) for two months and their NCS checked at the end of the treatment period. RESULTS: At least one electrophysiological parameter was abnormal in 38 out of 51 patients (74.6%). At the end of the treatment, Stage 1 a patients (NCS pathologic and neurologic examination normal) demonstrated a 44% improvement in all pathologic NCS parameters and a 50% improvement in sympathetic skin responses (SSR), while in Stage 1 b patients (NCS and neurologic examination pathologic) a matching ratio of improvement was detected in SSR but no definite improvement was noted in the all pathologic NCS parameters. CONCLUSION: Starting carnitine treatment in the early stages may be more effective in the treatment of sub-clinical neuropathy. A two-month treatment period may not be sufficient in detecting an electrophysiological improvement in cases where neurological deficits had been determined PMID- 16315972 TI - The effect of infrared laser on sensory radial nerve electrophysiological parameters. AB - OBJECTS: It has been claimed that laser may have bio-stimulation effect on the nerve tissues. This study has been designed to investigate the effect of different doses of infra-red (IR) laser exposure on the electrophysiological parameters of sensory nerves. METHODS AND SUBJECTS: Forty healthy subjects (20-35 years old) with no history of neurological conditions participated in this study. IR laser (780 nm, 20 mw) was applied over five blocks (1 cm2 each and 0.5 J/cm2) of 5 cm length of the left and right superficial radial nerve. The IR laser radiation was started from proximal to distal in the right hand and vise versa in the left hand. Antidromic sensory nerve conduction velocity was evaluated before and after first (0.5 J), third (1.5 J) and fifth (2.5 J) exposures. During the test, we measured the onset and peak latency, amplitude and duration of sensory action potentials. RESULTS: Paired t-test was used to assess the difference between pre- test and post- test data. After IR laser exposure with all doses, significant increased in latencies was observed (P < 0.001), while significant decreased in amplitude and duration was found only in the group who received the doses of 1.5 and 2.5 J of exposure (P < 0.001). There was no difference between right and left hands. CONCLUSION: Our results showed that the minimal dose of IR laser may not produce enough effects on the sensory nerves, while the higher doses such as 1.5 and 2.5 J may activate the mechanism of nerve blockage. PMID- 16315973 TI - Acute pure motor quadriplegia: is it dengue myositis? AB - BACKGROUND AND AIM: In view of paucity of comprehensive evaluation about dengue infection producing quadriplegia, we report the clinical, laboratory and neurophysiological studies in these patients. SUBJECTS AND METHODS: Seven out of 16 patients with dengue infection presented with quadriplegia and they were subjected to a detailed clinical history and examination. Diagnosis of dengue was based on characteristic clinical and positive serum IgM ELISA. Blood counts, serum chemistry, CSF analysis and nerve conduction and electromyographic (EMG) studies were performed in all. Outcome was defined at the end of 1 month into complete, partial and poor on the basis of activities of daily living RESULTS: The age of the patients ranged between 9 and 42 years and 2 were females. Fever was present in all and myalgia in 5 patients. Weakness developed within 3-5 days of illness, which was severe in 4 and moderate in 3 patients. Hypotonia and hyporeflexia were present in 5 patients. Nerve conduction and EMG studies were normal in all except one whose EMG was myopathic. Serum CPK and SGPT were raised in all and serum bilirubin in 3 patients. All the patients had coagulopathy and 6 had thrombocytopenia. Muscle biopsy in 1 patient was suggestive of myositis. Six patients improved completely and one had poor recovery who needed ventilatory support. CONCLUSION: Dengue virus infection may result in acute pure motor quadriplegia due to myositis. In an endemic area it should be considered in the differential diagnosis of acute flaccid paralysis. PMID- 16315974 TI - Widespread electromyographic abnormalities in patients with monomelic amyotrophy: a detailed EMG study. AB - OBJECTIVE: To evaluate subclinical electromyographic changes in unaffected muscles of the patients with monomelic amyotrophy (MMA). PATIENTS AND METHODS: 35 patients of MMA with single limb atrophy (30 patients with upper limb involvement and 5 patients with lower limb involvement) were studied at All India Institute of Medical Sciences, New Dellhi, from September 2000 to September 2002. All the patients were evaluated clinically, by detailed three limb electromyography (EMG) and by MRI scan of the spine to rule out other disorders. RESULTS: The mean age of 35 male patients was 24.17 (+/-6.8) years and the mean duration of illness was 3.64 (+/-2.7) years. Patients with upper limb involvement had segmental pattern of atrophy, predominantly distal or proximal. EMG revealed evidence of subclinical diffuse chronic reinnervative changes. All the patients (100%) had bilateral chronic reinnervative changes and 50% had chronic reinnervative changes in three limbs. CONCLUSIONS: Patients with clinically single limb MMA were found to have evidence of widespread chronic reinnervative changes on EMG. PMID- 16315975 TI - Phase and frequency coordination between neuron firing as an integrative mechanism of human CNS self-organization. AB - It was shown by surface electromyography (sEMG) from spinal cord injury and Parkinson's disease patients that FF-type motor units fire repeatedly at a rate of 10 Hz and FR-type motor units with approximately 3 action potentials per impulse train at 4 to 5 Hz. Such oscillatory firing was compared with the firing of alpha1, alpha2, and alpha3-motoneurons, innervating FF FR, and S-type muscle fibres, which fire oscillatory approximately at 10 Hz, 4 to 7 Hz and 1 Hz, as measured earlier with the single nerve-fibre action potential recording method from motoneuron axons. Repeated firings with impulse trains consisted of 1, 2-5, and 20-50 action potentials per impulse train respectively. Oscillatory firing of motoneurons has now been demonstrated in humans with two methods. In spinal cord injury patients it was observed that the FF-type motor units, firing normally at 10 Hz, fired for higher activations also at approximately 20 Hz. A motor program burst was generated by recruiting for the burst time motor units to fire oscillatory. It was further shown that FF-type motor units fired with relative phase and frequency coordination but non-synchronously. The coordination was between motor units firing of the same muscle and between muscles of the right and left arm. With the single-nerve fibre action potential recording method it was found earlier that alpha and gamma-motoneurons and muscle spindle afferents fired in relative coordination. There is indication that relative phase and frequency coordination is an integrative mechanism for the self-organization of the neuronal networks of the human central nervous system (CNS). PMID- 16315976 TI - [Thermophilic microbial metal reduction]. AB - Thermophilic microorganisms can reduce Fe(III), Mn(IV), Cr(VI), U(VI), Tc(VII), Co(III), Mo(VI), Au(I, III), and Hg(II). Ferric iron and Mn(IV) can be used as electron acceptors during growth; the physiological role of the reduction of the other metals is unclear. The process of microbial dissimilatory reduction of Fe(III) is the most thoroughly studied. Iron-reducing prokaryotes have been found in virtually all of the recognized types of terrestrial ecosystems, from hot continental springs to goethermally heated subsurface sediments. Thermophilic iron reducers do not belong to a phylogenetically homogenous group and include representatives of many bacterial and archaeal taxa. Iron reducing thermophiles can couple Fe(III) reduction with oxidation of a wide spectrum of organic and inorganic compounds. In the thermophilic microbial community, they can fulfil both degradative and productive functions. Thermophilic prokaryotes probably carried out global reduction of metals on Earth in ancient times, and, at the same time, they are promising candidates for use in modern biotechnological processes. PMID- 16315977 TI - [Dependence of the genotypic characteristics of Acidithiobacillus ferrooxidans on the physical, chemical, and electrophysical properties of pyrites]. AB - Comparison of Acidithiobacillus ferrooxidans strains TFV-1 and TFBk with respect to their capacity to oxidize pyrite 1, with hole-type (p-type) conductivity, or pyrite 2, with an electron-type (n-type) conductivity, showed that, at a pulp density of 1%, both before and after its adaptation to the pyrites, strain TFBk, isolated from a substrate with a more complex mineral composition, grew faster and oxidized the pyrites of both conductivity types more efficiently than strain TFV-1, which was isolated from a mineralogically simple ore. At a pulp density of 3-5%, the oxidation of pyrite 1 by strain TFV-1 and both of the pyrites by strain TFBk began only after an artificial increase in Eh to 600 mV. If the pulp density was increased gradually, strain TFBk could oxidize the pyrites at its higher values than strain TFV-1, with the rate of pyrite 2 oxidation being higher than that of pyrite 1. During chemical oxidation of both of the pyrites, an increase was observed in the absolute values of the coefficients of thermoelectromotive force (KTEMF); during bacterial-chemical oxidation, the KTEMF of pyrite 1 changed insignificantly, whereas the KTEMF of pyrite 2 decreased. PMID- 16315978 TI - [Dependence of the genotypic characteristics of Acidithiobacillus ferrooxidans on the physical, chemical, and electrophysical properties of pyrites]. AB - This study focused on the effect of physical, chemical, and electrophysical properties of two pyrites, pyrite 1, which had hole-type (p-type) conductivity, and pyrite 2, with electron-type (n-type) conductivity, on the genotypic characteristics of Acidithiobacillus ferrooxidans strains TFV-1 and TFBk, which were isolated from different substrates. After the adaptation of the strains to the pyrites at a pulp density of 1%, pulsed-field electrophoresis revealed changes in the chromosomal DNA of strain TFV-1 adapted to pyrite 1 and strain TFBk adapted to either of the pyrite types. In pyrite-adapted strain TFBk, the plasmid composition was the same as after growth on a medium containing ferrous iron, whereas, in strain TFV-1, changes in plasmid sizes or both in plasmid sizes and plasmid number occurred. After an increase in the density of the pyrite 2 pulp from 1 to 10%, the plasmid number increased from three to four, and, after an increase in the density of the pyrite 1 pulp from 1 to 7%, the plasmid number increased from two to six. PMID- 16315979 TI - [The effect of succinate on respiration, transamination, and pyruvate formation in cells of the yeast Dipodascus magnusii]. AB - The effect of succinate on the growth and respiration of the yeast Dipodascus magnusii VKM Y-1072, which is auxotrophic for thiamine and biotin, was studied. The addition of succinate to a culture grown on glucose was found to activate the respiration of cells on various substrates by enhancing the processes related to transamination reactions. In this case, aerobic fermentation (ethanol production) decreased, whereas pyruvate production increased. When succinate was added to the medium as the sole carbon source, it supported yeast growth in the absence of one of the two vitamins, thiamine or biotin, but not both. The yeast metabolism was completely respiratory, without any signs of aerobic fermentation. A drastic rise in pyruvate production in the yeast grown on glucose in the presence of succinate and the absence of biotin are also indicative of metabolic changes. PMID- 16315980 TI - [Mechanisms of interaction between DNA and chemical analogues of microbial anabiosis autoinducers]. AB - The alkylhydroxybenzene (AHB) autoregulatory factors d1 (fd1) of microorganisms have been found to directly interact with highly polymeric DNA. This circumstance results in changes, related to alterations in the topology of this macromolecule, in DNA physicochemical properties. The physicochemical properties of DNA in the presence of chemical analogues of microbial AHBs (methylresorcinol; hexylresorcinol; and 2-(4-hydroxyphenyl)ethane-1-ol, also known as tyrosol) were investigated using adsorption spectrophotometry, fluorometry, heat denaturation, viscosimetry, and electrophoresis in agarose gel. A number of concordant effects pointing to DNA-AHB interactions were revealed that manifesed themselves in the hypochromic properties of the resulting complexes, an increase in their melting temperature and viscosity, a decrease in their electrophoretic mobility, and a change in the fluorescent properties of AHBs upon complexation with DNA. Such alterations were particularly significant in the presence of hexylresorcinol, which possessed the maximum alkyl radical length among the fd1 analogues tested. Using atomic force microscopy, we visualized the micelle-like DNA nanostructures forming in the presence of AHBs. The results obtained provided the basis for developing a hypothetical model of the interaction between the biopolymer macromolecule and low-molecular-weight AHBs that takes into account the differences in the hydrophobicity of individual AHB homologues functioning as ligands. In terms of our model, we discuss AHB involvement in the stabilization of DNA and alteration of its topology, i.e., in the process related to intragenomic rearrangements, which account for the intrapopulational variability of bacteria, including dissociation processes. PMID- 16315981 TI - [Determination of the structure of the repeated unit of the Azospirillum brasilense SR75 O-specific polysaccharide and homology of the lps loci in the plasmids of Azospirillum brasilense strains SR75 and Sp245]. AB - The structural identity of the repeated unit in O-specific polysaccharides (OPSs) present in the outer membrane of strain SR75 of the bacterium Azospirillum brasilense, isolated from wheat rhizosphere in Saratov oblast, and the OPSs of previously studied A. brasilense strain Sp245, isolated from surface-sterilized wheat roots in Brazil, has been demonstrated. Plasmid profiles, DNA restriction, and hybridization assays suggested that A. brasilense strains SR75 and Sp245 have different genomic structures. It was shown that homologous lps loci of both strains was localized in their plasmid DNA. This fact allows us to state that, despite their different origin, the development of the strains studied was convergent. Presumably, the habitation of these bacteria in similar ecological niches influenced this process in many respects. PMID- 16315982 TI - [Characterization of lipopolysaccharides from Pseudomonas fluorescens IMB 2108 (biovar II) and IMB 2111 (biovar IV) with O-chains represented by alpha-glucan]. AB - Results of studies of the structurally unique O-chains of lipopolysaccharides, which were isolated from the dry biomass of Pseudomonas fluorescens IMB 2108 (biovar II) and IMB 2111 (biovar IV) by the Westphal technique and purified by repeated ultracentrifugation, are reported. The bulk of the lipopolysaccharide preparations contained S- and R-molecules at an average molar ratio of 1: 2. The main components of the hydrophobic moiety of lipid A were 3-hydroxydecanoic, 2 hydroxydodecanoic, 3-hydroxydodecanoic, dodecanoic, hexadecanoic, and octadecanoic acids, as well as hexadecenoic and octadecenoic acids. Glucosamine and phosphoethanolamine were identified as components of the hydrophilic moiety of lipid A. The degree of lipid A phosphorylation amounted to 3-4%. Fractions of the core oligosaccharide contained glucose, galactose, mannose, rhamnose, arabinose, glucosamine (only in strain IMB 2108), alanine, phosphoethanolamine, phosphorus, and 2-keto-3-deoxyoctulosonic acid (KDO). Heptose was present in trace amounts. O-specific polysaccharide chains were represented by a linear polymer of D-glucose units, which were linked together via alpha-(1,4) glycoside bonds. The existence of P. fluorescens strains that have alpha-1,4-glucan as the O-chain of their lipopolysaccharides has not been described before. PMID- 16315983 TI - [Clostridium alkalicellum sp. nov., an obligately alkaliphilic cellulolytic bacterium from a soda lake in the Baikal region]. AB - The first anaerobic alkaliphilic cellulolytic microorganism has been isolated from the Verkhnee Beloe soda lake (Buryatiya, Russia) with pH 10.2 and a salt content of up to 24 g/l. Five strains were characterized. Strain Z-7026 was chosen as the type strain. The cells of the isolate are gram-positive spore forming rods. A mucous external capsule is produced. The microorganism is obligately alkaliphilic, growing in a pH range of 8.0-10.2, with an optimum at pH 9.0. Sodium ions and, in carbonate-buffered media, sodium chloride are obligately required. The microorganism is slightly halophilic; it grows at 0.017-0.4 M Na+ with an optimum at 0.15-0.3 M Na+. The metabolism is fermentative and strictly anaerobic. Cellulose, cellobiose, and xylan can be used as growth substrates. Plant and algal debris can be fermented. Lactate, ethanol, acetate, hydrogen, and traces of formate are produced during cellulose or cellobiose fermentation. Yeast extract or vitamins are required for anabolic purposes. The microorganism fixes dinitrogen and is nitrogenase-positive. It is tolerant to up to 48 mM Na2S. Growth is not inhibited by kanamycin or neomycin. Chloramphenicol, streptomycin, penicillin, ampicillin, ampiox, bacillin, novobiocin, and bacitracin suppress growth. The DNA G+C content is 29.9 mol %. According to the nucleotide sequence of its 16S rRNA gene, strain Z-7026 is phylogenetically close to the neutrophilic cellulolytic bacteria Clostridium thermocellum (95.5%), C. aldrichii (94.9%), and Acetivibrio cellulolyticus (94.8%). It is proposed as a new species: Clostridium alkalicellum sp. nov. PMID- 16315984 TI - [Description of Desulfotomaculum nigrificans subsp. salinus as a new species, Desulfotomaculum salinum sp. nov]. AB - This study focused on the physiological, chemotaxonomic, and genotypic characteristics of two thermophilic spore-forming sulfate-reducing bacterial strains, 435T and 781, of which the former has previously been assigned to the subspecies Desulfotomaculum nigrificans subsp. salinus. Both strains reduced sulfate with the resulting production of H2S on media supplemented with H2 + CO2, formate, lactate, pyruvate, malate, fumarate, succinate, methanol, ethanol, propanol, butanol, butyrate, valerate, or palmitate. Lactate oxidation resulted in acetate accumulation; butyrate was oxidized completely, with acetate as an intermediate product. Growth on acetate was slow and weak. Sulfate, sulfite, thiosulfate, and elemental sulfur, but not nitrate, served as electron acceptors for growth with lactate. The bacteria performed dismutation of thiosulfate to sulfate and hydrogen sulfide. In the absence of sulfate, pyruvate but not lactate was fermented. Cytochromes of b and c types were present. The temperature and pH optima for both strains were 60-65 degrees C and pH 7.0. Bacteria grew at 0 to 4.5-6.0% NaCl in the medium, with the optimum being at 0.5-1.0%. Phylogenetic analysis based on a comparison of incomplete 16S rRNA sequences revealed that both strains belonged to the C cluster of the genus Desulfotomaculum, exhibiting 95.5-98.3% homology with the previously described species. The level of DNA-DNA hybridization of strains 435T and 781 with each other was 97%, while that with closely related species D. kuznetsovii 17T was 51-52%. Based on the phenotypic and genotypic properties of strains 435T and 781, it is suggested that they be assigned to a new species: Desulfotomaculum salinum sp. nov., comb. nov. (type strain 435T = VKM B 1492T). PMID- 16315985 TI - [Isolation of bacteria of the family enterobacteriaceae from plant tissues]. AB - Bacteria of the family Enterobacteriaceae were isolated from the tissues of a number of wild and cultivated plants. All the cultures isolated had a broad spectrum of resistance to antibiotics and were highly adhesive to human erythrocytes. The studies conducted indicate the possibility of concentration of microorganisms pathogenic for humans in plant tissues. PMID- 16315986 TI - [Characterization of communities of heterotrophic bacteria associated with healthy and diseased corals in Nha Trang Bay (Vietnam)]. AB - A comparative investigation of the heterotrophic microflora of 11 species of healthy corals and of white-band-diseased and yellow-band-diseased corals inhabiting the reefs of Nha Trang Bay (Vietnam), which has been exposed to anthropogenic impact, was performed. Fifty-nine strains of heterotrophic bacteria isolated on Y/K and Endo media were investigated and characterized. All the isolates were identified at the genus level by consideration of the results of analysis of their phenotypic properties, determination of the molar percent of G+C bases in their DNA, and the composition of fatty acids of their lipids. In the composition of the microflora of tissues of healthy corals, gamma proteobacteria prevailed, with halomonads being dominant among them. In addition, the gram-negative bacteria included Pseudomonas and Vibrio spp., members of the Cytophaga-Flavobacterium-Bacteroides (CFB) phylogenetic cluster, and Moraxella sp. The gram-positive bacteria revealed included Bacillus, Staphylococcus, Halococcus, and Micrococcus spp., and coryneform bacteria. In the composition of the microflora of the tissues of affected corals, bacteria of the family Enterobacteriaceae and of the genera Planococcus and Arthrobacter, which were not revealed in healthy hydrobionts, were found. The anthropogenic impact is not the sole factor determining the infection of corals. PMID- 16315987 TI - [Seasonal changes in the structure of the anoxygenic phototrophic bacterial community in Lake Mogilnoe, a relict lake on Kil'din Island in the Barents Sea]. AB - An anaerobic phototrophic bacterial community in Lake Mogilnoe, a relict lake on Kil'din Island in the Barents Sea, was studied in June 1999 and September 2001. Irrespective of the season, the upper layer of the anaerobic zone of this lake had a specific species composition of sulfur phototrophic bacteria, which were dominated by the brown-colored green sulfur bacterium Chlorobium phaeovibrioides. The maximum number of phototrophic sulfur bacteria was observed in June 1999 at a depth of 9 m, which corresponded to a concentration of bacteriochlorophyll (Bchl) e equal to 4.6 mg/l. In September 2001, the maximum concentration of this pigment (3.4 mg/l) was found at a depth of 10 m. In both seasons, the concentration of Bchl a did not exceed 3 microg/l. Purple sulfur bacteria were low in number, which can be explained by their poor adaptation to the hydrochemical and optical conditions of the Lake Mogilnoe water. In June 1999, the water contained a considerable number of Pelodictyon phaeum microcolonies and Prosthecochloris phaeoasteroides cell chains, which was not the case in September 2001. A 16S rDNA based phylogenetic analysis of pure cultures of phototrophic bacteria isolated from the lake water confirmed that the bacterial community is dominated by Chl. phaeovibrioides and showed the presence of three minor species, Thiocvstis gelatinosa, Thiocapsa sp., and Thiorhodococcus sp., the last of which is specific to Lake Mogilnoe. PMID- 16315988 TI - [Geographical races of certain species of ascomycetous yeasts in the Moscow and Novosibirsk regions]. AB - Strains of three species of the ascomycetous yeasts Hanseniaspora guilliermondii, Torulaspora delbrueckii, and Debaryomyces hansenii, isolated from the above ground parts of plants in similar biocenoses of distant geographic regions (Moscow and Novosibirsk regions), have been investigated. The strains in each species were indistinguishable with respect to phenotypic features and general DNA characteristics as determined by restriction analysis. However, comparison of the strains using PCR analysis with nonspecific primers revealed considerable intraspecific variability. From their electrophoretic patterns, the strains of the three species studied were found to cluster in accordance with the region of isolation. This phenomenon is interpreted as an example of the existence of geographical races in the major eurytopic species of yeasts. PMID- 16315989 TI - [Succession of chitinolytic microorganisms in chernozem soil]. AB - The chitinolytic prokaryotic and eukaryotic microbial complex of chernozem soil has been investigated in the course of a succession initiated by the introduction of chitin and humidification. The dynamics of the cell numbers of chitinolytic microorganisms and of their biomass was assessed by fluorescent microscopy and by inoculation of selective media. Emission of carbon dioxide and nitrous oxide, as well as dinitrogen fixation, was assessed by gas chromatography. It was found that, when the succession was initiated by the introduction of both chitin and humidification, it resulted in greater cell numbers and biomass of chitinolytic microorganisms and higher levels of CO2 and N2O emission and of nitrogen fixation than when the succession was initiated by humidification alone. As compared to the control samples, a significant (twofold) increase in the prokaryote cell number and biomass was found on the fourth day of the succession initiated by humidification and introduction of chitin. One week after the initiation of succession, the fungal biomass and length of mycelium were twice as high as those in the control samples. These results led to the conclusion that chitin utilization in chernozem soil starts during the initial stages of succession and is performed by both prokaryotic and eukaryotic microorganisms. PMID- 16315990 TI - [Methane oxidation in landfill cover soil]. AB - Methane oxidation in the cover soil of the Khmet'evo municipal landfill in Moscow oblast was investigated. Methane emission from the experimental parcel of the site was highly inhomogeneous. At a depth of 45-60 cm, the pore gas mainly consisted of CH4 (60-70%) and CO2 (30-40%). In the upper horizons of the cover soil, the concentration of these gases sharply decreased. Techniques for estimation of the methane-oxidizing activity in the cover soil of the landfill were tested. The rate of methane oxidation in the soil, the factor limiting methane emission from the surface of the site, correlated with the cell number of culturable methanotrophic bacteria. The method of indirect immunofluorescence revealed ten known species of methanotrophic bacteria in enrichment cultures obtained from samples of the cover soil. Our results also indicate the presence of unknown psychrotolerant methanotrophs that are active at the low temperatures characteristic of Moscow oblast. PMID- 16315991 TI - [Methanol and glucose metabolism in Beijerinckia mobilis]. PMID- 16315992 TI - [The role of succinate dehydrogenase in bacterial resistance to the delta endotoxins of Bacillus thuringiensis]. PMID- 16315993 TI - [Relationship between the carbohydrate specificity of lectins and the carbohydrate composition of the Lentinus edodes (Berk.) Sing [Lentinula edodes (Berk.) Pegler] mycelium at different stages of its morphogenesis]. PMID- 16315994 TI - [Interrelation of the biosyntheses of lipids, lipoxygenase, and lipase in cultured streptomycetes]. PMID- 16315995 TI - [Photodynamic therapy in age-related macular degeneration]. AB - INTRODUCTION: Exudative form of age-related macular degeneration is the leading cause of legal blindness in elderly. PURPOSE: The purpose of this study is to report 3.5 years experience with photodynamic therapy in this disorder. PATIENTS AND METHODS: Patient files of all patients underwent photodynamic therapy with verteporfin in Department of Ophthalmology, Semmelweis University, Budapest in a 3.5 years period were reviewed. Changes in visual acuity were considered as main outcome variable compared to natural history data. RESULTS: From April, 2000 to September 2003, 302 patients were treated using photodynamic therapy with verteporfin for exudative form of age-related macular degeneration. The mean change of visual acuity were 2.6 and 2,7 lines at 12 and 24 months, respectively, while according to the natural history data 4.2 and 4.5 could have been expected. After 2 years, 83% of the patients had visual acuity better than or equal to 20/200 (this is the level of legal blindness), while only 33% was expected according to natural history data. Visual acuity of 20/80 or better (practical ambulatory vision) was found in 39% of the treated eyes (12% in natural history data). There was no severe adverse event. CONCLUSIONS: Photodynamic therapy was found to reduce the risk of severe visual loss. The authors' results confirm the data from the international trials. To achieve good results, good patient compliance and continuous access to the treatment are indispensible. PMID- 16315996 TI - [Genetics of primary biliary cirrhosis]. AB - The primary biliary cirrhosis is a chronic cholestatic liver disease, which is characterised by non-suppurative destruction of interlobular bile ducts. The precise etiopathogenesis of primary biliary cirrhosis remains unknown. Evidence suggest that genetic and environmental factors seem to be important. It shows strong heritability according to familial occurrence and monozygotic twins concordance. There is an increase in the degree of monosomy of the X chromosome in female subjects with PBC. There is an association with HLA-DR8 (DR1*08) antigen at least in some populations. Correlation the PBC with polymorphisms of HLA class I, II, III alleles, genes encoding for molecules influencing immune tolerance, apoptosis, cytokine expression, will require additional studies. PMID- 16315997 TI - [Clinical significance of subchorionic and retroplacental hematomas detected in the first trimester of pregnancy]. AB - AIMS: To evaluate the long-term clinical significance of intrauterine hematomas detected in the first trimester of pregnancy in a general obstetric population. METHODS: A prospective study was designed to compare the perinatal outcome in 187 pregnant women with intrauterine hematomas to 6488 controls in which hematomas were not detected at first trimester by ultrasound examination. RESULTS: The incidence of intrauterine hematoma in the first trimester in a general obstetric population was 3.1%. A retroplacental position of the hematoma was significantly correlated with an increased risk for adverse maternal and neonatal complications. The presence or absence of symptoms of threatened abortion did not affect these outcomes. The rates of operative vaginal delivery (RR: 1.9; CI: 1.1 3.2) and cesarean section (RR: 1.4; CI: 1.1-1.8) were significantly greater in the hematoma group as compared to the control group, as well as the rates of pregnancy induced hypertension (RR: 2.1; CI: 1.5-2.9) and preeclampsia (RR: 4.0; CI: 2.4-6.7). Placental abruption (RR: 5.6; CI: 2.8-11.1), and the incidence of placental separation abnormalities was also significantly more frequent in the hematoma group (RR: 3.2; CI: 2.2-4.7). Perinatal complications, including the rate of preterm delivery (RR: 2.3; CI: 1.6-3.2), intrauterine growth restriction (RR: 2.4; CI: 1.4-4.1), fetal distress (RR: 2.6; CI: 1.9-3.5), meconium stained amniotic fluid (RR: 2.2; CI: 1.7-2.9), and NICU admission (RR: 5.6; CI: 4.1-7.6) were also significantly increased in this group. Furthermore, the frequency of intrauterine demise and perinatal mortality was increased in the hematoma group, but this difference did not reach statistical significance (p = 0.6 and p = 0.2). CONCLUSION: The authors' study suggests that the presence of an intrauterine hematoma during the first trimester may identify a population of patients at increased risk for adverse pregnancy outcome. PMID- 16315998 TI - [Biparietal diameter/kidney length ratio in cases with chronic hypoxic state]. AB - AIMS: The object of this study was to investigate the fetal biparietal diameter/kidney length ratio in normal and hyperechogenic kidneys during the third trimester of gestation. MATERIAL AND METHODS: The selected cases were characterized by pregnancy-associated hypertension and/or proteinuria, which was associated with fetal hypoxia due to the depression of placental blood flow. Depending on the renal manifestation of intrauterine chronic hypoxia, cases were divided into two study groups. Group I was composed of 21 fetuses with pregnancy associated hypertension and/or proteinuria and hyperechogenic renal medullae. Group II consisted of 162 fetuses with pregnancy-associated hypertension and/or proteinuria and normal echoic kidney. Both study groups included pregnant women from the third trimester. RESULTS: Fetal renal hyperechogenicity correlated with the pathological growth of fetal kidney. The fetal biparietal diameter/kidney length ratio was significantly lower in cases of hyperechogenicity. CONCLUSIONS: The fetal renal hyperechogenicity is a relevant indicator of diminution of fetal renal perfusion. This can lead to abnormal development of the affected kidney and can result in a pathological reduction of biparietal diameter/kidney length ratio, which may also be an in utero indicator of subsequent intrauterine and neonatal complications. Detailed ultrasound examinations of renal parenchyma and kidney length seem to be a useful method in the prenatal diagnosis of decreased renal perfusion and of intrauterine hypoxia and serve to detect pathological conditions in utero. PMID- 16315999 TI - [Lajos Markusovszky Memorial Meeting of Orvosi Hetilap, 2005]. PMID- 16316000 TI - [Breast, body constitution, fashion and breast cancer]. PMID- 16316001 TI - [Base excision repair: AP endonucleases and DNA polymerases]. AB - The DNA base lesions in living cells occur permanently and with high frequency as a result of the action of exogenous and endogenous factors. The main mechanism providing removal of such lesions is base excision repair. PMID- 16316002 TI - [Genetic collection of meiotic mutants of rye Secale cereale L]. AB - Genetic collection of meiotic mutants of winter rye Secale cereale L. (2n = 14) was created. Mutations were detected in inbred F2 generations after self fertilization of the F1 hybrids, obtained by individual crossing of rye plants (cultivar Vyatka) or weedy rye with plants from autofertile lines. The mutations cause partial or complete plant sterility and are maintained in collection in a heterozygous state. Genetic analysis accompanied by cytogenetic study of meiosis has revealed six mutation types. (1) Nonallelic asynaptic mutations sy1 and sy9 caused the formation of only axial chromosome elements in prophase and anaphase. The synaptonemal complexes (SCs) were absent, the formation of the chromosome "bouquet" was impaired, and all chromosomes were univalent in meiotic metaphase I in 96% (sy1) and 67% (sy2) of cells. (2) Weak asynaptic mutation sy3, which hindered complete termination of synapsis in prophase II. Subterminal asynaptic segments were always observed in the SC, and at least one pair of univalents was present in metaphase I, but the number of cells with univalents did not exceed 2%. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19, which caused partially nonhomologous synapsis: change in pairing partners and fold-back chromosome synapsis in prophase I. In metaphase I, the number of univalents varied and multivalents were observed. (4) Mutation mei6, which causes the formation of ultrastructural protrusions on the lateral SC elements, gaps and branching of these elements. (5) Allelic mutations mei8 and mei10, which caused irregular chromatin condensation along chromosomes in prophase I, sticking and fragmentation of chromosomes in metaphase I. (6) Allelic mutations mei5 and mei10, which caused chromosome hypercondensation, defects of the division spindle formation, and random arrest of cells at different meiotic stages. However, these mutations did not affect the formation of microspore envelopes even around the cells, whose development was blocked at prophase I. Analysis of cytological pictures of meiosis in double rye mutants reveled epistatic interaction in the mutation series sy9 > sy1 > sy3 > sy19, which reflects the order of switching these genes in the course of meiosis. The expression of genes sy2 and sy19 was shown to be controlled by modifier genes. Most meiotic mutations found in rye have analogs in other plant species. PMID- 16316003 TI - [The uncertainty of "fitness:" what prevents understanding of the role of genetic exchange]. AB - The evolutionary development of highly organized species is attained through an increase in average survival of individuals, whereas the evolution of primitive species involves only an increase in fecundity (Zavadsky, 1958, 1961). However, in population genetics, survival (or ecological resistance) and fecundity are regarded as components of a single character, fitness. Employment of the notion of fitness, which lacks a strict definition, hinders understanding of the mechanism of progressive evolution as the process that enhances ecological resistance of organisms. The notion of fitness also exacerbates understanding the role of genetic exchange, since the primary advantage of genetic recombination and sexual reproduction apparently is producing of progeny with high ecological resistance rather than with high genetic diversity as such. Thus, the regular genetic exchange ensures restoration of the level of ecological resistance characteristic for the species, and on the macroevolutionary scale leads to the formation of new genomes and new species with high ecological resistance. PMID- 16316004 TI - [Secretion of biologically active human granulocyte colony-stimulating factor (G CSF) in milk of transgenic mice]. AB - Two constructs were devised, containing the full-length gene of the human granulocyte colony-stimulating factor (G-CSF) fused with the 5' and 3' flanking promoter sequences of bovine alpha-S1-casein gene. Both constructs contained a 1518-bp fragment that included exons 18 and 19 and 320 bp of the 3' flanking region of bovine gene @CSN1S1, but differed in size of the 5' flanking sequences, which were of 721 bp, and exon 1 in construct pGCm1 and 2001 bp and exon 1 and intron 1 in construct pGCm2. With both constructs, transgenic mice were produced. The transgene expression was assessed using RT-PCR and immunochemically from the production of human G-CSF in milk of lactating females. Secretion of human G-CSF into the milk varied in a wide range, from 0.8 microg/ml to over 1 mg/ml, in mice with construct pGCml and was low (up to 60 microg/ml) or absent in mice with construct pGCm2. G-CSF glycosylation was incomplete in mice with transgene pGCml and complete in mice with pGCm2. G-CSF of transgenic mouse milk was shown to stimulate the formation and growth of granulocyte-containing colonies in human umbilical blood cell culture and be close or identical in physiological activity to the natural human G-CSF. PMID- 16316005 TI - [A2144G is the main mutation in the 23S rRNA gene of Helicobacter pylori associated with clarithromycin resistance]. AB - To detect point mutations A2115C, A2143G/C, and A2143G in the 23S rRNA gene of Helicobacter pylori associated with resistance of the microorganism to clarithromycin, a new powerful way of analysis was used. This method involved the reaction of minisequencing followed by MALDI-TOF mass spectrometry of reaction products. In ten analyzed clarithromycin-resistant clinical isolates of H. pylori obtained in Russia, the resistance was found to be mediated only by mutation A2144G in the 23S rRNA gene. PMID- 16316006 TI - [Genetic diversity of Mongolian gazelle Procapra guttorosa Pallas, 1777]. AB - The mitochondrial DNA D-loop hypervariable fragment sequence polymorphism was examined in 27 Mongolian gazelles from Mongolia, Russia, and China. Intraspecific polymorphism of the D-loop fragment examined was demonstrated. All haplotypes described were unique. The average nucleotide diversity (pi) for the mtDNA fragment investigated constituted 5.85 +/- 2.92%. A relatively high number of insertions and deletions was observed. In particular, a haplotype with the 77-bp insertion was described. The data obtained point to high genetic diversity of Mongolian populations. There was no correlation between the distribution of haplotypes examined and geographical location of the animal tissue sampling sites. PMID- 16316007 TI - [Visualization of chromosome territories in interphase nuclei of ovarian nurse cells in Calliphora erythrocephala Mg. (Diptera: Calliphoridae)]. AB - Analysis of localization of chromosomes 2, 3, and 6 of Calliphora erythrocephala Mg. in ovarian nurse cell nuclei with different chromatin structure has shown that the regions of DNA probe hybridization reduced with increasing chromatin compaction. Hybridization of DNA probes of chromosomes 3 and 6 to secondary reticular nuclei demonstrated that chromosomes retain their territories in the nuclei when the chromatin acquires a reticular structure. These results suggest regular organization of the chromosomal apparatus at all stages of the endomitotic cycle, including the stage of highly polyploid reticular nuclei. FISH of DNA probe of the chromosome 2 telomeric region to secondary reticular nuclei revealed a peripheral distribution of the signal. Zones of more intensive DNA probe hybridization have been distinguished. These zones probably are the regions of accumulation of telomeric and (or) centromeric chromosome regions. PMID- 16316008 TI - [Analysis of genetic diversity of spring durum wheat (Triticum durum Desf.) cultivars released in Russia in 1929-2004]. AB - Based on genealogical analysis, the genetic diversity of 78 spring durum wheat cultivars released in Russia in 1929-2004 have been examined. The temporal trends of change in diversity were studied using series of n x m matrices (where n is the number of the cultivars and m is the number of original ancestors) and calculating coefficients of parentage in sets of cultivars released in particular years. The pool of original ancestors of spring durum wheat cultivars includes 90 landraces and old varieties, more than a half (57%) of which originate from European countries, including Russia and Ukraine (45%). The original ancestors strongly differ in the frequency of presence in the cultivar pedigrees. Landraces Beloturka, Sivouska, Kubanka (T. durum Desf.), Transbaikalian emmer, Yaroslav emmer (T. dicoccum Schuebl.), Poltavka (T. aestivum L.), and the original ancestors of cultivars Kharkov 46, Narodnaya, and Melanopus 1932 enter in the pedigrees of more than half of cultivars created within the framework of various breeding programs. At that, their distribution by cultivars from different breeding centers strongly varies. Analysis of temporal dynamics of genetic diversity, based on genetic profiles and coefficients of parentage, has shown that the genetic diversity of Russian durum wheats increased during the period examined. Nevertheless, genetic erosion of the local material-a loss of approximately 20% of the pool of Russian original ancestors-has been found. The contribution of the original ancestors to the pedigrees of different cultivars, constructed in different breeding centers and recommended for cultivation in different regions, has been estimated. The variation of the released cultivars was highest in the Lower Volga region and lowest in the Ural region. In all, the lower threshold of genetic diversity in all regions does not reach the critical level, corresponding to the similarity of half-sibs. The set of modern cultivars included in the Russian Official List 2004 has a cluster structure. PMID- 16316009 TI - [Variability in size of the nuclear genome in pygmy wood mouse Sylvaemus uralensis (Rodentia, Muridae)]. AB - Earlier, in an integral genetic study, the Asian and European races were distinguished within the species Sylvaemus uralensis (pygmy wood mouse) and the European race was divided into the East European and South European forms. Each of these groups differed from the others, in particular, in the quantity of the centromeric heterochromatin in karyotypes of the animals. To establish the pattern of its changes in S. uralensis, in the present study the DNA content in splenocyte nuclei in all races and forms of pygmy wood mice was assessed using DNA flow cytometry. The heterochromatin amount in karyotypes and genome size were shown to be correlated. The East European chromosomal race of S. uralensis (Central Chernozem and Non-Chernozem regions of Russia, Crimea Peninsula, Middle Volga region, and Southern Ural) and the Asian race of this species (East Kazakhstan, Uzbekistan, and East Turkmenistan), which have respectively the highest and the lowest amounts of centromeric heterochromatin in the karyotype, exhibit the greatest difference in the DNA content in the genome. On average, the difference is approximately 8% in males and 6.7% in females; in both cases, the ranges of variability were distinctly different. Against the general background of the trait variation, the Asian race, whose members have the smallest DNA amount in their cells, looks homogeneous. The genome of the South European chromosomal form of S. uralensis (Caucasus, Transcaucasia, Carpathians, and Balkan Peninsula), which exhibits an intermediate content of the centromeric heterochromatin in the karyotype, is smaller that the genome of the East European race (by 3.2% in the group of males and by 1.9%, in the group of females), but larger than that of the Asian race (by 5% in either sex). Thus, the variability of size of centromeric C-blocks in pygmy wood mouse is likely to be associated with elimination (or, conversely, an increase in the amount) of the genetically inert chromatin. It is suggested that a significant contribution to the variability of genome size in S. uralensis is made by heterochromosomes, or, more precisely, their variable regions, which seem to be largely heterochromatic. PMID- 16316010 TI - [The effect of mutation dominant spotting-Yurlovo (KitW-Y) on spermatogenesis, early embryogenesis, and fertility of C57BL/6JY mice]. AB - The effect of mutation KitW-Y found in C57BL/6 mice on fertility, spermatogenesis, and early embryogenesis of mice have been studied. If heterozygotes KitW-Y/+ are crossed with wild-type mice, fertility decreases by 20%. Homozygotes Kitw-Y/KitW-Y and compounds KitW-Y/KitSsm are nonviable. The study of spermatogenesis in KitW-Y/+ mice has demonstrated a negative effect of this mutation on spermatocytes. Histological examination of the testes of mutant males has shown local empty spaces in seminal ducts. Electron microscopic examination of synaptonemal complexes have demonstrated desynapsis disturbance in some nuclei at the diplotene stage of meiotic prophase I. However, these disturbances do not cause a decrease in the number of fertilized oocytes/ova. The decrease in fertility is accounted for disturbances of early embryogenesis. In vivo and in vitro analyses of early embryogenesis have demonstrated that cleavage divisions are asynchronous in KitW-Y/+ heterozygous embryos. Some of these embryos die before implantation, and others cleave more rapidly than wildtype embryos, which give them selective advantage during the postimplantation period of embryogenesis. The pattern of KitW-Y expression during spermatogenesis and embryogenesis mimics potential human pathology, which makes these mutants an interesting and valuable object for genetics and developmental biology. PMID- 16316011 TI - TGFalpha reactivates imprinted Igf2 in the parthenogenetic mice embryos and placenta. AB - Imprinted genes play important roles in the mammalian development. In the parthenogenetic embryos (PE) there is only expression of maternally expressed genes. Therefore, PEs are appropriate experimental models to study genomic imprinting controlling mechanisms. The maternally expressed H19 and paternally expressed Igf2 are reciprocally imprinted genes in normal embryos. Here we studied effect of transforming growth factor alpha (TGFalpha) treatment in vitro (10 ng/ml at the morula stage) on the expression of Igf2/H19 locus in mice PE (9.5-days of gestation, 25 somites) and their placentas (PP). Using RT-PCR we showed that TGFalpha reactivated maternally imprinted Igf2 gene in parthenogenetic embryos and placentas. In spite of similar Tgfalpha expression in the pre-implantation stages, its expression in the 9.5-day parthenogenetic embryos is significantly less than in normal embryos (NE). In our experiments it was shown that reactivation of Igf2 gene occurred independently of H19 gene. In vitro TGFalpha treatment of mouse PE reactivated paternally expressed Igf2 gene in the PE and PP. In the PE and PP both Igf2 and H19 were expressed. It seems that TGFalpha can play an important role as modulator of the Igf2/H19 locus. PMID- 16316012 TI - [Genetic demographic structure of the Lugansk (Ukraine) population: age, ethnicity, education, and occupation]. AB - Data from marriage records from registry office archives of 1960, 1985, 1990, and 2000 have been used to calculate the genetic demographic parameters of the Lugansk city population. The population is mostly Ukrainian and Russian. In the years studied, Ukrainians accounted for 49.4, 44.0, 44.6, and 48.7% of the population and Russians, for 46.0, 52.1, 50.7, and 45.6%, respectively. The total proportion of the two ethnic groups decreased from 97.0 to 94.6% for 40 years. The marriage contingency coefficient (K) with respect to ethnicity decreased from 0.16 to 0.07 in the period between 1960 and 1985 and increased from 0.10 to 0.24 between 1990 and 2000. The frequency of interethnic marriages (48.4, 50.9, 52.0, and 60.0%, respectively, in the years studied) indicate that the outbreeding component has been constantly increasing in the Lugansk population. The marriage assortativeness indices (A') for Ukrainians were 10.2, 4.8, 2.1, and 6.0% in 1960, 1985, 1990, and 2000, respectively. For pooled minor ethnic groups, A' values in these years were 25.6, 12.4, 18.9, and 6.6%, respectively. The marriage contingency coefficient (K) with respect to occupation increased in the period between 1960 and 1990 (0.14, 0.16, and 0.19 in 1960, 1985, and 1990, respectively), and K with respect to education decreased (0.35, 0.30, and 0.26, respectively). PMID- 16316013 TI - [Cytogenetic analysis of spermatozoa in a patient with chromosome constitution 45,X/46,X,r(Y) by intracytoplasmic injection into mouse oocytes]. AB - The chromosome set of human spermatozoa was studied by intracytoplasmic injection into mouse oocytes. A total of 85 metaphase plates of male pronuclei of a patient with chromosome constitution 46,X,r(Y)/45,X and 108 metaphase plates of patients with normal sperm parameters (control group) were examined. The ratio between X- and Y-bearing chromosomes in the 46,X,r(Y)/45,X patient and in the control group did not differ from 1:1. A significant increase in the rates of diploidy, hypoploidy, hyperploidy of sex chromosomes, and chromosome structure rearrangements in spermatozoa of the patient in comparison with spermatozoa in the control group was recorded. PMID- 16316014 TI - [The population gene pool of Yakutia: cartographic analysis based on the polymorphism of immune and biochemical markers]. AB - The gene pool of the indigenous population of Sakha Republic (Yakutia) has been studied within the borders of this republic coinciding with the main area of Yakuts, which was formed by the end of the 19th century and have remained stable until the present time. Maps of the geographic variation of the integrated characteristics of the Yakut gene pool, including the principal components, parameters of genetic diversity, and genetic distances from the "average" Yakut population are presented. It has been demonstrated that ethnographers' reports on intense internal assimilation in modem Yakutia agree with genetic data. The stratification of the Yakut gene pool reflected in the maps of two principal components corresponds to the observed general (H(T)) and interpopulation (FST) gene diversities. PMID- 16316015 TI - [Relationship between the reproductive capability of a subdivided population and its genetic structure: computer simulation, the simplest multilocus model]. AB - An original computer model, simulating joint genetic and demographic dynamics of subdivided populations, is proposed. The model accounts for the reverse effect of the genetic structure on the reproductive capability of a population, which is based on a postulated limited set of biallelic loci, controlling variation in an adaptive quantitative trait. The model allows to simulate spreading of the population, originating from a single small colony, which involves genetic and demographic equilibrium (a normal population-genetic process) and reorganization of the genetic structure of the subdivided population upon anthropogenic pressure, associated with a decrease in its reproductive capability (an adverse population-genetic process). PMID- 16316016 TI - [Genetic analysis of anthocyanin of the anthers and culm pigmentation in common wheat]. AB - Anthocyanin pigmentation of various organs develops during plant ontogeny in response to adverse and damaging abiotic and biotic stressors (environmental factors). Using the monosome method, the genes responsible for anther and culm anthocyanin pigmentation (Pan1 and Pc2, respectively) were localized to 7D chromosome in introgressive lines from crosses between common wheat Triticum aestivum L. and the species Triticum timopheevii Zhuk. Genetic analysis of ten common wheat genotypes using testers carrying genes Pan1, Pn1, and Pn2 showed that these genotypes contained Pan1 and Pn2 genes. Visual examination of plants from 70 and 76 varieties of respectively winter and spring common wheat revealed anthocyanin pigmentation of anthers and culms in 36 varieties. Pan1 and Pn2 genes were presumably introduced into common wheat from Aegilops tauschii (Fig.) Tzvel., a donor of the D genome. PMID- 16316017 TI - [Population identification of Helicobacter pilory isolates from Russia]. AB - Using multilocus sequence typing (MLST), 22 Helicobacter pylori isolates from Russia have been characterized. All of the Russian strains were assigned to a single population, hpEurope. PMID- 16316018 TI - Presence of depolarization-induced suppression of inhibition in a fraction of gabaergic synaptic connections in rat neocortical cultures. AB - Brief depolarization of postsynaptic neurons in hippocampus and cerebellum results in a transient depression of GABAergic inhibitory input, called "depolarization-induced suppression of inhibition" (DSI). We studied whether a similar phenomenon occurs in the rat neocortical neurons. Using patch-clamp technique in neocortical cell cultures we examined the effects of a 5-second depolarization of postsynaptic neurons on evoked GABAergic inhibitory post synaptic currents (IPSCs). We found that the depolarization evoked a suppression of IPSC amplitude in 6 out of 26 neuronal pairs tested. The suppression of IPSC amplitude lasted for approximately 70 seconds and was accompanied by changes of paired-pulse ratio and IPSC coefficient of variation (CV), which is suggestive of a presynaptic mechanism. These results are in agreement with previous observations in hippocampal cell cultures and suggest that neocortical neurons express DSI. PMID- 16316019 TI - [Human sleep in terms of individual typological features]. AB - The authors consider sleep of healthy adults in terms of the theory of the integrated individuality and assume that the basic parameters of sleep architecture vary depending on gender, somatotype and psychophisiological features. Recent literature concerned with individual differences in sleep organization is reviewed, and the possible mechanisms of the distinctions are justified. PMID- 16316020 TI - [A role of the basal ganglia in the occurrence of visual hallucinations (a hypothetical mechanism)]. AB - A hypothetical mechanism of the basal ganglia involvement in visual hallucinations is proposed. According to this mechanism, hallucination is the result of modulation of the efficacy of corticostriatal synaptic inputs and changes in spiny cell activity due to the rise of striatal dopamine concentration (or due to other reasons). These changes cause an inhibition of neurons in the substantia nigra pars reticulata and subsequent disinhibition of neurons in the superior colliculus and pedunculopontine nucleus (including its cholinergic cells). In the absence of afferentation from the retina this disinhibition leads to activation of neurons in the lateral geniculate nucleus, pulvinar and other thalamic nuclei projecting to the primary and highest visual cortical areas, prefrontal cortex, and also back to the striatum. Hallucinations as conscious visual patterns are the result of selection of signals circulating in several interconnected loops each of which includes one of above mentioned neocortical areas, one of thalamic nuclei, limbic and one of visual areas of the basal ganglia, superior colliculus and/or pedunculopontine nucleus. According to our model, cannabinoids, opioids and ketamine may lead to hallucinations due to their promotional role in the LTD of cortical inputs to GABAergic spiny cells of striatal striosomes projecting to dopaminergic neurons, disinhibition of the lasts, and increase in striatal dopamine concentration. PMID- 16316021 TI - [The dynamics of the direct current potentials of the brain during sustained attention in 10-11-year-old children]. AB - The dynamics of direct current potentials of the brain was studied in 10-11-year old children during sustained attention to successive presentation of series of Shulte tables. Children were examined twice: before and after the series of training to fast reading. A gradual increase in the level of direct current potentials during sustained attention was observed. The increase was more pronounced in children with excessive than in children with moderate reactions to the loading. After the series of training to fast reading, the increase in the level of direct current potentials was reduced in both groups. This aftertraining neurophysiological phenomenon was combined with a transformation of psychophysiological characteristics: a decrease in the time of viewing of Shulte tables and increase in the speed of reading. It is suggested that the shifts of direct current potentials reflects the dynamics of intensity of the cerebral energy metabolism. PMID- 16316022 TI - [Assessment of recognition of emotions by healthy persons and patients with focal brain pathology]. AB - To assess the involvement of different structures of the human brain into successive stages of the recognition of the principal emotions by facial expression, we examined 48 patients with local brain lesions and 18 healthy adult subjects. It was shown that at the first (intuitive) stage of the recognition, premotor areas of the right hemisphere and temporal areas of the left hemisphere are of considerable importance in the recognition of both positive and negative emotions. In this process, the left temporal areas are substantially involved into the recognition of anger, and the right premotor areas predominantly participate in the recognition of fear. In patients with lesions of the right and left brain hemispheres, at the second (conscious) stage of recognition, the critical attitude to the assessment of emotions drops depending on the sign of the detected emotion. We have confirmed the hypothesis about a correlation between the personality features of the recognition of facial expressions and the dominant emotional state of a given subject. PMID- 16316023 TI - [EEG correlates of the change in information processing strategy in visual imagery]. AB - Cnanger of the spatial organization of biopotentials (spatial synchronization and disorder, spectral power and coherence) were analyzed during mental creating of visual images from two simple elements: the angle the oblique line. With the transition from the first to the fourth task, the total number of used elements increased from the number suitable for simultaneous presentation and conscious processing (less than 7 +/- 2) to a much higher number. Changes in the characteristics of the spatial organization of biopotentials associated with the increase in the number of the elements can be explained by a change in the information processing strategy, transformation of information processing strategy, i.e. the transition from the left-hemispheric successive (conscious) analyses to the right-hemispheric simultaneous (unconscious) processing. It was shown that this change in the information processing strategy was accompanied by an increase in the index of spatial disorder sensitive to complicated nonlinear processes. Subjects were divided in two groups with different forms of the reorganization of interhemispheric and fronto-occipital relationships of biopotentials. These data are interpreted in terms of different involvement of the unconscious intellectual processes of different layers (subconsciousness and superconsciousness) in the change in information processing strategy. PMID- 16316024 TI - [The role of the cognitive context in the conservatism of unconscious visual sets]. AB - Rigidity of a verbal set was compared in three series of experiments: (1) pseudowords were presented at the set-forming stage and were changed for common words at the testing stage; (2) in the same conditions as in 1, an additional task of target localization in the matrix by two distinctive features was introduced; (3) in the additional target localization task, the spatial component was strengthened whereas image recognition component was reduced. The results confirmed our hypothesis about the context-dependence of the rigidity of the visual set. This characteristic substantially depends on a working memory loading and cognitive tasks performed by a subject, in particular, the relationship between the degrees of involvement of the ventral and dorsal visual systems into the cortical processing of sequential verbal and nonverbal visual stimuli. The experimental paradigm can serve as a model for the investigation of the roles of the ventral and dorsal visual systems in the recognition function. PMID- 16316025 TI - [Asymmetry of the amplitude-temporal properties of visually-guided saccades in monkeys depending on the complexity of the spatial scheme of the visual stimulation]. AB - The properties of visually-guided saccades were studied in three monkeys (Macaca rhesus). Traditional single-step GAP-OVERLAP temporal paradigms were used. Target stimuli were presented in accordance with two spatial stimulation schemes: (i) along the horizontal meridian (one-dimensional scheme) and (ii) within rectangle part of the visual field (two-dimensional scheme). We revealed asymmetric focuses of short- and long-latency saccades in the two-dimensional scheme of stimulation. The MANOVA revealed that the factor of dimensionality influenced saccadic latency to a greater extent than the factor of lateralization of stimuli (presenting in the right or left visual hemifields). Accuracy of saccades decreased with increasing in their eccentricity in both spatial schemes of stimulation. PMID- 16316026 TI - [Individual typological features of the dog behavior during the choice between food and avoidance of electrocutaneous stimulation]. AB - Two conditioned reflexes (CR) on flash were elaborated in five dogs in three experimental series. In one of them pressing on pedal were constantly reinforced with two pieces of liver, in other--escape or avoidance of skin stimulation with lifting of leg. In a third series, series of reinforcement choice, in front of animal with electrodes, fasting on leg, put the pedal. During of choice series defensive reactions were dominated in two dogs, alimentary reactions--three of them. For restoration alimentary behaviour in dog with great dominating of defensive reactions were many times reinforced with--food "passive" pressing legs on pedal during conditional stimuli, dog with less dominating of these reactions for restoration were enough a few coupling of conditional stimuli with food. With intensification electrocutaneus stimulation of legs or decreasing alimentary reinforcement we a success of diminishing of degree of alimentary reaction domination in one dog and in other ones temporarily changed of these domination on prevalence of defensive reactions. The results were compared with data, obtained in these dogs with technique of choice between probability and value of alimentary reinforcement. This comparison were permitted us of ranging dogs on scale "carefulness-riskiness", revealed of individual typological features of these animals. PMID- 16316027 TI - [Regional features of electrical reactions (in the frequency band of 1-225 Hz) in the cerebral cortex to conditioned stimuli in the course of instrumental conditioning]. AB - Power spectra of short-term (less than 1 s) EEG-reactions (in the frequency band of 1-225 Hz) were studied in dogs in the course of instrumental food conditioning. These reactions were observed in different areas of the cortex in response to positive and differentiated conditioned stimuli. Regional features between the spectra were found both in the power level and frequency structure. The power of the reactions in the visual and parietal areas of the left hemisphere was higher than in the motor areas. Power spectra of reactions to differentiated stimuli were significantly lower than the spectra of reactions to positive stimuli mainly owing to the high-frequency components (80-225 Hz). In these both cases, prestimulus power spectra did not differ. The frequency structure of corresponding EEG-reactions consisted of individual spectral peaks, mainly both gamma (30-80 Hz) and higher-frequency (80-225 Hz) bands. PMID- 16316028 TI - [Effect of neurotensin on the behavior of rats with damaged serotoninergic neurons under immobilization stress]. AB - It was shown that the immobilization of animals has led to reducing of vertical and horizontal locomotor activity in the "open field" and decreasing of number of conditioned food-procuring reactions into T-maze. The damages of serotoninergic neurons produced via local injections of selective neurotoxin 5, 7 dihydroxytriptamine into dorsal raphe nucleus intensified behavior alterations. Neurotensin administrations reduced effects of neurotoxin: the rats locomotor activity and quantity of conditioned reactions into T-maze were kept at the phone level just after immobilization as well as next two days. The results indicate the important protective significance of neurotensinergic brain structures for ensuring of adaptive behavior of animals with damaged serotoninergic neurons under emotional stress conditions. It is supposed that neurotensin normalizing influences on behavior is connected to a restoration of balance of dopamine-and serotoninergic brain structures interaction. PMID- 16316029 TI - [Effect of vibrissae removal on the defensive behavior of rats in early ontogeny]. AB - Influence of restricted sensory afferentation in rats forming of defensive reactions was studied by vibrissectomy from 9 to 20 day of postnatal ontogeny. Defensive withdrawal reaction in response to touch from 10 to 18 day, duration of freezing on 20 day, "open field" behavior on 25 day were measured. Intensity of withdrawal reactions, freezing duration, flight reaction and emotionality in "open field" were lower in vibrissectomized rats with comparison to control. PMID- 16316030 TI - [Modulation of the basal forebrain neural activity related to efficiency of reaching for food movements in rat]. AB - In the course of learning, single unit activity of basal forebrain neurons was recorded in freely moving rats. Modulation of the neural activity related to efficiency of reaching for food movements was observed between a 40% of the recorded neurons. The firing-rate of such neurons increased during retrieval of the food. Conversely, the activity of these neurons decreased or did not change during unsuccessful movements. In the most neurons (57.1%) the modulation of the activity was observed at the second half of time interval of the movement, but before finishing its. Some neurons (35.7%) changed their activity after the movements. PMID- 16316031 TI - [Conditioning in resuscitated rats with originally different behavioral patterns]. AB - Male white rats divided in three groups on the basis of their behavior in the elevated plus-maze were subjected to 10-min cardiac arrest. Analysis of the results of their subsequent food and active avoidance conditioning revealed the effects of the following factors: (1) a factor of initial typological features of the higher nervous activity, (2) a factor of general brain mechanisms of postresuscitation, and (3) a factor of postresuscitation features in rats with different behaviors, which represented changes in different mechanisms of conditioned reflex performance in these animals. PMID- 16316032 TI - [Recovery of motor skills after the caudate lesion in rats with the different prefered forelimb: the role of intense retraining]. AB - Rats were trained for instrumental reaching (17 mm) of a sunflower seed from a horizontal tube 12 mm in diameter. After training, rats were divided into groups, "right-" and "left-handers", by the forelimb preference. Unilateral electrolytic lesions of the head of the caudate nucleus contralateral to the prefered forelimb were performed. After surgery, animals were retrained to perform the food retrieval reaction by the same forelimb (reaching reaction by the "intact" forelimb was prohibited by a special bracelet). Both right- and left-handers were divided into groups of rare and intense retraining. Over the course of five months, animals of the groups of rare training were tested once a week, whereas the intense retraining was performed three or four times a week. Rats with right and left forelimb preference were shown to recover the reaching skill with different rates. In general, animals with left-side caudate lesion (right handers) recovered the skill better both under condition of spontaneous recovery (rare testing) and intense retraining. The results suggest different mechanism of skill recovery after the right- and left-side brain lesion. PMID- 16316033 TI - [Crossintervalograms for analysis of electrical activity of the brain]. AB - A method of construction of crossintervalograms for electroencephalograms is proposed. Particular points or EEG fragments of particular shape are used as time locked events, and the intervals between these reference points are analyzed. The method is theoretically substantiated. Examples of crossintervalograms constructed for EEG extrema and derivative EEGextrema are given. The information validity of these crossintervalograms is demonstrated and their features are indicated. Comparison crossintervalograms with crosscorrelograms is performed. The possibilities of further development and applications of a method are specified. It is suggested that the method will be useful for investigation of operative interaction of brain subsystems. PMID- 16316034 TI - [Use of the peptide strategy to study mechanisms of transferring the hormone signal to the cell]. PMID- 16316035 TI - [Difference in the gene expression of antibacterial peptides abaecin, hymenoptaecin, defensin in bees Apis mellifera and Apis mellifera caucasica]. PMID- 16316036 TI - [Comparative study of fluoroorganic ammonium reversible inhibitors of cholinesterases in various animals]. PMID- 16316037 TI - [Evolutionary conservative and species specific antigenic determinants of in mammals thyroglobulins]. PMID- 16316038 TI - [Effect of hypoxia on physiological and biochemical blood values in sea fishes]. PMID- 16316039 TI - [Acetylcholinesterase activity in the olfactory bulb of Esox lucius and its role in the development of the long posttetanic potentiation]. PMID- 16316040 TI - [Cardioelectric field on the body surface of birds at the activation of the atrial myocardium]. PMID- 16316041 TI - [Locomotion disorders in the progeny of ambidextrous rat females with a psychogenic trauma]. PMID- 16316042 TI - [Placenta type interaction and evolutionary trends of the uterus development in the Cestoda]. PMID- 16316043 TI - [Projection of olfactory bulbs and non-olfactory structures of the telencephalon in the amygdalar complex of the turtle Testudo horsfieldi: study using anterograde tracer technique]. PMID- 16316044 TI - [Three dimensional arrangement of the AB2 type epithelia using bird cochlear epithelium as an example]. PMID- 16316046 TI - [Study of evolutionary physiology at the Petersburg State University]. PMID- 16316047 TI - Sixth meeting of the Technical Advisory Group on the Global Elimination of Lymphatic Filariasis, Geneva, Switzerland, 20-23 September 2005. PMID- 16316048 TI - [The fifty-year old whole body autoradiography]. AB - In this paper the method of whole body autoradiography is reviewed. Methods of obtaining sections and visualization of the latent images are described. Features of detection methods are compared and the evaluation methods of autoradiograms are given. The usage of the procedure in the pre-clinical stage of drug development is illustrated by means of a few examples obtained in our laboratory. PMID- 16316049 TI - [Application of metal ions and their complexes in medicine II. Application of platina complexes in the treatment of tumor]. AB - The rapid development of inorganic medical chemistry opens enormous potential for various applications of a range of inorganic substances in the medicine. Thus inorganic chemistry offers real possibilities to pharmaceutical industries, which used to be dominated by organic chemistry alone. The field has particularly been stimulated by the success-story of cisplatin, which is the World's best selling anticancer drug. Nowadays orally administered Pt(IV) complexes with reduced toxicity, and activity against resistant tumors are on various phases of clinical trial. PMID- 16316050 TI - [HPLC investigation of antioxidant components in Solidago herba]. AB - Representatives of Solidago species have been used in European phytotheraphy for centuries as a component of urological and antiphlogistical remedies. Solidago canadensis L. (Asteraceae) contains a wide range of active ingredients, such as flavonoids, saponins, hydroxycinnamates and mineral elements, which are responsible for its characteristic anti-inflammatory, spasmolytic and diuretic properties. Quality control of collected Solidaginis herba were performed according to the instructions of the X. German Pharmacopoea, while different LC MS technologies were applied to evaluate the exact phenoloid composition. Three flavonol aglycons (quercetin, kaempferol and isorhamnetin) connected to several sugar components (glucose, rhamnose, galactose and rutinose), caffeoylquinic acid and a caffeoyl-shikimic acid glycoside were identified in the samples. Quercetin 3-O-beta-glucoside (isoquercitrin), quercetin-3-O-beta-galactoside (hyperoside), quercetin-3-O-beta-rhamnoside (quercitrin), quercetin-3-O-beta-rutinoside (rutin), kaempferol-3-O-beta-rhamnoside (afzelin), kaempferol-3-O-beta-rutinoside (nicotiflorin), caffeoil-quinic acid (chlorogenic acid) were identified in sample "A", while the presence of quercetin, quercetin-3-O-beta-glucoside (isoquercitrin), quercetin-3-/6"-O-acetyl-/-beta-glucopiranoside, quercetin-3-O beta-rutinoside (rutin), kaempferol, kaempferol-3-O-beta-glucoside (astragalin), kaempferol-3-/6"-O-acetyl-/-beta-glucopiranoside, isorhamnetin, isorhamnetin-3 /6"-O-acetyl-/-beta-glucopiranoside, isorhamnetin-3-O-beta-rutinoside (narcissin), caffeoil-quinic acid (chlorogenic acid), caffeoil-shikimic acid glucoside (dattelic acid-glucoside) were confirmed in sample "B". According to the occurrence of acetyl-glycosides and the diversity of sugar component of flavonoid glycosides Solidaginis herba samples chemotaxonomically were classified into different varieties. Incidence of acetyl-glycosidic flavonoids and absence of flavonoid galactosides and rhamnosides in the sample "B" together give support for the taxonomic recognition of varietases Solidago canadensis L. var. canadensis and var. scabra. Sample "A" was identified as Solidago canadensis L. var. canadensis, while sample "B" has proved to be belong to variety Solidago canadensis L. var. scabra. Due to the same flavonoid aglycons and the large amounts of flavonol glycosides occurring in each drug, phytochemical characteristics of investigated samples proved to be very similar. PMID- 16316051 TI - [Utilization study of clozapine, olanzapine and risperidone in Hungary]. AB - Authors examined the use of clozapine, olanzapine and risperidone by reviewing retrospectively their prescribing practice in the period of 1999-2003 in Hungary. The aim of the study was to analyze the utilization aspects of the use of the three novel antipsychotics. Data showed a fast growing trend in the use of olanzapine and risperidone while the use of clozapine is decreasing. The purpose of the present study was to follow the temporal development trends of use of the three examined novel antipsychotics which could result in a cost-effective drug therapy. PMID- 16316052 TI - [Pharmaceutical chemistry of sedato-hypnotics and anxiolytics]. AB - The paper represents the 2nd part of a series about agents acting on the central nervous system. As previously, the material is divided into chapters of hystory, preparation; structure-properties-activity; therapeutical use; analysis. PMID- 16316053 TI - [Some aspects of intraspecific variability of the closely related species of the Dermacentor marginatus complex (Acari: Ixodidae) as demonstration of microevolutionary process]. AB - Individual and geographical variability of male conscutum pattern in five closely related species of the Dermacentor marginatus complex (D. marginatus, D. ushakovae, D. niveus, D. silvarum, D. nuttalli) has been examined. The pattern has been examined in three to six geographical samples within each specific distribution range. It was established that the pattern of the conscutum central area is subjected to individual and geographical variability on an extreme degree and gives very few intraspecific differential characters, but it shows interspecific differences in some cases. However the dark patches of marginal flank (npkappadelta and 3deltakappa respectively, see Fig. 3) show statistically significant differences in length in all geographical samples of the first four species. Characteristic features of ixodid ticks morpogenesis rule out to a considerable extent the possibility to trace geographical variability of the same characters by all stages of ontogenesis. In these cases, when it is possible, the tendencies to variability on different stages do not always coincide. There is no coordination within the species in the degree of differences of examined geographical samples by pattern and morphological characters, i. e. very complex polymorphis is observed. In the context of the evolution that kind of polymorphism of the geographical samples has provided the species with high adaptive potential for conquest of the extensive range and maintaining large numbers. The question of allopatric speciation of the closely related D. marginatus-D. silvarum with small secondary area of transgression their ranges, and sympatric speciation of the D. ushakovae-D. niveus is postulated. PMID- 16316054 TI - [The peculiarities of the biology of ticks inhabiting the environs of Tomsk City]. AB - The presence of ticks Ixodes persulcatus and I. pavlovskyi occidentalis is revealed in urban parks and the forests directly adjoined to buildings. I. pavlovskyi occidentalis is found to be the dominant species nearby the city. The both species are occurred almost simultaneously, and the peak of their abundance usually falls on 2nd or 3rd ten-day period of May. In the city the ticks species ceased occurring in registers in the end of May, and in the adjacent forests they disappeared in the end of June. I. pavlovskyi occidentalis has not been found in the forests remote from the city at the distance more than 15 kilometers. Besides the above species the ticks Dermacentor reticulatus and Haemaphysalis concinna are occurred rarely in the territory examined. PMID- 16316055 TI - [Six new species of microsporidia of the genus Amblyospora (Microspora: Amblyosporidae) from blood sucking mosquitoes (Diptera: Culicidae) from the west Siberia]. AB - Microsporidia of the genus Amblyospora parasiting the adipose body of mosquito larvae of the genus Aedes and Culex has been studied with both light and electron microscopy. Six new species of microsporidia are described based on ultrastructural characteristics of spores and sporogony stages. Amblyospora flavescens sp. n. Mature spores are egg-shaped. The spore wall with three layers, about 165 nm. Exospore is two-membranous. Subexospore is absent. Endospore is electron-translucent. Polaroplast consists of three parts: lamellar, large vesicular, lamellar. The anisofilar polar filament with 10--11 coils (3 1/2 + 2 1/2 + 4-5). Fixed spores are 6.3 +/- 0.1 x 4.24 +/- 0.1 microm. Amblyospora kolarovi sp. n. Mature spores are egg-shaped. The spore wall with three layers, about 265-315 nm. Exospore shapes tucks on the surface of spore. It is two membranous. Subexospore is quagge, structural. Endospore is electron-translucent. Polaroplast consists of two parts: lamellar and large vesicular. The anisofilar polar filament with 11-13 coils (3 + 8-10). Fixed spores are 5.4-5.6 x 3.5-4.2 microm. Amblyospora orbiculata sp. n. Mature spores are widely egg-shaped. On a back pole there is a small concavity. The spore wall with three layers, about 155 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is absent. Endospore is electron-translucent. Polaroplast consists of three parts: lamellar, vesicular, lamellar. Polar filament is anisofilar, with 11 1/2 coils (4 1/2 + 1 + 6). Fixed spores are 6.3 +/- 0.1 x x 4.0 +/- 0.1 microm. Amblyospora rugosa sp. n. Mature spores are egg-shaped. On a back pole there is a small concavity. The spore wall with three layers, about 225 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is quaggy, structural. Endospore is electron-translucent. Polaroplast lamellate. Polar filament is anisofilar, with 17 1/2 coils (3 1/2 + 1 + 13). Fixed spores are 5.3 +/- 0.1 x 3.7 +/- 0.1 microm. Amblyospora undata sp. n. Mature spores are egg shaped. The spore wall is three-layered, about 220 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is quaggy, structural. Endospore is electron-translucent. Polaroplast lamellate. The anisofilar polar filament with 8 coils (3 + 5). Fixed spores are 5.0 +/- 0.1 x 3.0 +/- 0.1 microm. Amblyospora urski sp. n. Mature spores have widely oval form. The back pole is concave. The spore wall with three layers, about 280 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is quaggy, structural. Endospore is electron-translucent. Polaroplast lamellate. Polar filament is anisofilar, with 6 coils (2 + 4). Fixed spores are 4.4 +/- 0.1 x 2.9 +/- 0.1 microm. PMID- 16316056 TI - [Phenotypic variability of a perch parasite-cestode proteocephalus percae (Muller, 1780) (Proteocephalidea) in different parts of the species range]. AB - Discrete variability of four P. percae characters of the main cestode functional complexes was identified. Phenotypic diversity of P. percae from different parts of the distribution range was analysed. Research revealed low geographic variability and high stability of the dominant variations in combination with morphometric plasticity. The conclusion was made that the patterns of P. percae morphological variability were shaped by the common fate and long-standing co evolutionary relations between the parasite and the host--the perch Perca fluviatilis. PMID- 16316057 TI - [Influence of the phase of the number and demographic structure of the water vole population on its infection by helminthes]. AB - In this work we have analyzed results of the long-term investigations of the helminthes distribution in the various demography groups of the water vole population (Arvicola terrestris L.) in North Baraba (Novosibirsk region). The data on the dominant parasites of these rodents: trematodes Notocotylus noyeri (Joyeux, 1922), cestodes Limnolepis transfuga Spassky et Merkuscheva, 1967, nematodes Capillaria wioletti Ruchljadeva, 1950, Longistriata minuta (Dujarden, 1845) and Heligmosomum costellatum (Dujarden, 1845) have shown that the number of parasites in biocenosis are connected with different factors, such as the demographic structure of the host population, the alternation of hosts number and conditions of the environment (dry and damp phases of the climatic cycle). In the dry phase the main parasitize load N. noyeri, L. transfuga and C. wioletti connects with the breeding group; in the damp period - with immature rodents. Independently of the phase climatic cycle and the density of the water vole population the great bulk of nematodes L. minuta and H. costellatum was uncovered of the immature individuals. In any case the number of helminthes changed synchronously with such of the its host. PMID- 16316058 TI - [The influence of infections with trematodes Bunocotyle progenetica (Hemiuridae) and Cryptocotyle cancavum (Heterophyidae) onto mortality of littoral mollusks Hydrobia ulvae (Gastropoda: Prosobranchia) after freezing]. AB - The mortality of prosobranch snails Hydrobia ulvae uninfected and infected with the parthenites of two species of Trematodes (Bunocotyle progenetica and Cryptocotyle cancavum) after freezing in seawater of different salinity with following thawing has been studied. Reduced salinity exceeds mortality of snails under freezing significant (p < 0.01). The mortality of mollusks increases significant (p < 0.01) with the increase of the exposure time. The mortality of females exceeds the mortality of male significant (p < 0.01) within the group of 2-years individuals. The negative effect of trematode invasion to the vitality of mollusks after freezing was not showed. Trematode infestation registered in March 2004 and in September 2003 is not significant differ (Levakin, 2004). Thus any important differential mortality of individuals H. ulvae infected with parthenites of this Trematodes is not present during a winter time. The conclusion is conformed by the results of our experiments and well accords to features of realization of life cycles of these Trematodes under conditions of the White Sea intertidial zone, where infection is possible only within the worm season. PMID- 16316059 TI - [Patterns of host-parasite interactions between the nematode Heligmosomum mixtum (Schulz, 1952) and the bank vole (Clethrionomys glareolus Schreber, 1780)]. AB - Some aspects of the host-parasite interaction of the nematode Heligmosomum mixtum and the bank vole have been studied. The dependence of infestation on the host sex, age and weight, the seasonal and annual abundance dynamics are investigated. It has been found that the bank vole is the main host of H. mixtum; the distribution of H. mixtum abundances in the host population is described by a negative binomial distribution model. The greatest infestation is characteristic for mature bank voles; males are infected more heavily than females. Infestation with the nematode increases alongside with the host weight. Seasonal dynamics of the nematode abundance in a bank vole population appears as a curve with a maximum in January and a minimum in August; the infestation curve for 1-month-old animals captured from June through October has two peaks (in July and October), with a minimum in June. The number of parasites in the ecosystem over a long-term period changed synchronously with the host abundance. The synchronism in the host parasite system dynamics is possible when the parasite has a short life span, and does not influence the host abundance. PMID- 16316060 TI - [Trematoda of the genus aporocotyle (Sangyinicolata: Aporocotylidae) from fish far eastern seas]. AB - According to our research there are two species of trematoda of genus Aporocotyle: A. simplex and A. theragrae in the far eastern seas. PMID- 16316061 TI - [A study of possible transovarial and transphase transmission of borreliae by the tick Dermacentor reticulatus (Ixodidae)]. AB - A study of possible transovarial and transphase transmission of borreliae by the tick Dermacentor reticulatus is carried out. The possibility of borreliae transmission by the ticks of this species being infected spontaneously as well as experimentally is shown in principle. Although borreliae are preserved in the organism of D. reticulatus, low values of the infestation are established in the different stages of D. reticulatus development and in all steps of the study. D. reticulatus may be involved in the process of borreliae circulation in natural foci of tick-born borreliosis, but this species has no a significant importance for the maintenance of it. PMID- 16316062 TI - [The distribution of the urban mosquitoes, Culex pipiens pipiens f. molestus (Diptera: Culicidae) in Petrozavodsk City]. AB - The distribution of the urban mosquitoes, Culex pipiens pipiens f. molestus in Petrozavodsk city was studied for the first time. Immature mosquito stages were found in 21 of 68 basements flooded with polluted waters, the occurrence of the mosquitoes varied in different districts of the sity. The mean siphonal index of the mosquito larvae from 6 underground populations varied from 3.8 up to 4.2, i. e. within the range typical for C. p. p. f. molestus. PMID- 16316063 TI - HIV testing in the United States, 2002. AB - OBJECTIVE: This report presents national estimates of testing for Human immunodeficiency virus, or HIV, the virus that causes acquired immunodeficiency syndrome (AIDS). The objectives are to present nationally representative estimates of the degree of self-reported lifetime and recent HIV testing among persons 15-44 years of age in the United States. The report also contains data on sources of testing, reasons for tests, and whether HIV counseling was obtained. METHODS: Data from the 2002 NSFG, conducted by the Centers for Disease Control and Prevention (CDC), National Center for Health Statistics (NCHS), are based on interviews with a national sample of the household population of the United States. In-person, face-to-face interviews were conducted in the homes of 12,571 males and females 15-44 years of age in 2002. Most of the data used for this report were collected by an interviewer who asked the questions and entered the answers into a laptop computer. RESULTS: One-half of men and women 15-44 years of age in 2002 reported that they had been tested at least once (other than through blood donation), and 15.1 percent had been tested in the past 12 months, which is equivalent to 17-20 million tests per year among 15-44 year olds. Testing is more common in some population subgroups than others, for example, among African Americans and persons with increased risk for HIV. Private physicians and HMOs were the largest provider of tests, accounting for 45 percent of recent tests. Public sources accounted for 22 percent of tests. A minority of recently tested respondents (29 percent) reported talking with a health professional about the HIV test after being tested. Among women who had recently been pregnant, 69 percent reported being tested for HIV during prenatal care. Persons 15-44 years of age with increased risk for HIV, defined by drug-related or sex-related behavior, had higher reported testing during their lifetime and in the past 12 months than those not at higher risk. However, one-third of this higher risk group reported that they had never had an HIV test, equivalent to 4.1-5.5 million untested, at-risk persons aged 15-44 years, and a majority of higher risk persons had not been tested in the past year. PMID- 16316064 TI - Pharmaceutical development of a parenteral lyophilised dosage form for the novel anticancer agent C1311. AB - C1311 (5-[[2-(diethylamino)ethyl]amino]-8-hydroxyimidazo [4,5,1-de]-acridin-6-one dihydrochloride trihydrate) is the lead compound from the group of imidazoacridinones, a novel group of rationally designed anticancer agents. C1311 shows significant cytotoxic activity in vitro and in vivo toward a range of colon tumours. The aim of the present study is to develop a sterile and stable, injectable pharmaceutical product for C1311 to be used in phase I clinical trials. C1311 drug substance was structurally and analytically characterised by chromatographic, spectrometric, and diffraction techniques. C1311 was freely soluble in water, and its stability was investigated in several liquid and lyophilised formulations with or without the use of buffering, tonicity, and bulking agents. The final product, containing 100 mg/vial C1311 (as anhydrous free base), was stable for at least 3 months under accelerated storage conditions and at the designated long-term storage condition of 5 +/- 3 degrees C in the dark. The drug is currently used in phase I clinical trials. PMID- 16316065 TI - Optimization of the freeze-drying cycle: adaptation of the pressure rise analysis model to non-instantaneous isolation valves. AB - The principal aim of this study is to extend to a pilot freeze-dryer equipped with a non-instantaneous isolation valve the previously presented pressure rise analysis (PRA) model for monitoring the product temperature and the resistance to mass transfer of the dried layer during primary drying. This method, derived from the original MTM method previously published, consists of interrupting rapidly (a few seconds) the water vapour flow from the sublimation chamber to the condenser and analysing the resulting dynamics of the total chamber pressure increase. The valve effect on the pressure rise profile observed during the isolation valve closing period was corrected by introducing in the initial PRA model a valve characteristic function factor which turned out to be independent of the operating conditions. This new extended PRA model was validated by implementing successively the two types of valves and by analysing the pressure rise kinetics data with the corresponding PRA models in the same operating conditions. The coherence and consistency shown on the identified parameter values (sublimation front temperature, dried layer mass transfer resistance) allowed validation of this extended PRA model with a non-instantaneous isolation valve. These results confirm that the PRA method, with or without an instantaneous isolation valve, is appropriate for on-line monitoring of product characteristics during freeze drying. The advantages of PRA are that the method is rapid, non-invasive, and global. Consequently, PRA might become a powerful and promising tool not only for the control of pilot freeze-dryers but also for industrial freeze-dryers equipped with external condensers. PMID- 16316066 TI - Statistical properties of the USP dissolution test with pooled samples. AB - The Montecarlo simulation method is used to study the statistical properties of the USP pooled dissolution test. In this paper, the statistical behavior of the dissolution test for pooled samples is studied, including: a) the operating characteristic curve showing the probability of passing the test versus the mean amount dissolved, b) the influence of measurement uncertainty on the result of the test, c) an analysis of the dependence of the statistical behavior on the underlying distribution of the individual amounts dissolved, d) a comparison of the statistical behavior of the unit dissolution test versus the pooled dissolution test, e) the average number of stages needed to reach a decision presented as a function of parameters of the lot, f) the relative influence of the three stages of the test on the probability of acceptance. PMID- 16316067 TI - Impact of Blow/Fill/Seal process variables in determining rate of vial contamination by air dispersed microorganisms. AB - Controlled challenges of air dispersed spores of Bacillus subtilis NCIMB 8649 have been generated in a custom-built challenge room housing a Blow/Fill/Seal machine filling filter-sterilized trypticase soy broth into 5.5 cm3 low density polyethylene vials. The effects on the rate of vial contamination of systematic changes in the process variables, rate of provision of ballooning air, delay in the application of mould vacuum and duration of transfer of the open vial, have been examined. Overall, the findings show that the conditions of vial formation can affect appreciably the rate of vial contamination from airborne spores. The indications are that heat lethality, associated with the elevated temperature required for polymer extrusion and vial formation, has a role in determining such contamination. PMID- 16316068 TI - Simple HPLC determination of benzalkonium chloride in ophthalmic formulations containing antazoline and tetrahydrozoline. AB - A simple and rapid analytical procedure for routine quantification of n-C12H25 and n-C14H29 benzalkonium chloride (C-12 and C-14 BKC) homologs in ophthalmic formulations containing antazoline HCl and tetrahydrozoline HCl by high performance liquid chromatography was developed and validated. The ophthalmic solution samples can be directly analyzed by reversed-phase on HiQ-Sil C18 column (4.6 mm x 150 mm, i.d., 5-microm particle size) with acetonitrile-sodium acetate buffer (pH 5.0; 0.2 M) (70:30, v/v) as mobile phase. UV Detection was carried out at 262 nm. The method was linear over the selected concentration and ranged from 0.03 to 0.10 mg/ml (r2 = 0.9999) and from 0.01 to 0.05 mg/ml (r2 = 0.9979) for C 12 and C-14 BKC homologs, respectively. The mean percent recoveries were 100.2 and 102.6 and the percent CV values were 1.3 and 3.5 for C-12 and C-14 BKC homologs, respectively. The results demonstrated the good linearity, accuracy, and precision. The method was applied to determine two commercial ophthalmic formulations, and the percent label amounts of total BKC contents were found to be 99.7 and 103.2. PMID- 16316069 TI - PDA technical report no. 42: Process validation of protein manufacturing. Parenteral drug Association. PMID- 16316070 TI - [Ego strength level and treatment for alcoholics]. AB - In this study, the level of ego strength between alcoholics and normal controls were compared by means of response obtained from TAT (Thematic Apperception Test) Card 1. Further, the ego strength level of alcoholics were classified into four Groups (A, B, C, and D). The characteristics of these groups were compared in terms of factors such as the ego strength level, the frequency in use of denial, the motivation for a treatment and the participation rate to a self-help group and the treatment effects. PMID- 16316071 TI - [Effects of ethanol ingestion on glucose tolerance in people with different genotypes of ALDH2]. AB - Effects of alcohol drinking on glucose tolerance were investigated in male healthy subjects (20-36 years old) with different phenotypes of aldehyde dehydrogenase 2 (ALDH2). Oral glucose tolerance test was performed in each subject twice with and without simultaneous oral ingestion of ethanol. Simultaneous oral ingestion of ethanol (10 g) significantly enhanced the early increase in plasma insulin concentration at 30 min after ingestion of glucose (100 g) in subjects with normal ALDH2 phenotype, while the increase in early insulin response due to alcohol was slight and not significant in those with atypical ALDH2. On the other hand, the early increase in plasma glucose concentration at 30 min after glucose inges-tion was not affected by simultaneous ethanol drinking in both groups of subjects with normal and atypical ALDH2 phenotypes. Thus, drinking alcohol together with a simple sugar causes enhancement of early insulin response, which is more prominent in people with normal ALDH2 phenotype than in those with atypical ALDH2 phenotype. PMID- 16316072 TI - [Social networks in drinking behaviors among Japanese: support network, drinking network, and intervening network]. AB - PURPOSE: The national representative sample was analyzed to examine the relationship between respondents' drinking practice and the social network which was constructed of three different types of network: support network, drinking network, and intervening network. METHOD: Non-parametric statistical analysis was conducted with chi square method and ANOVA analysis, due to the risk of small samples in some basic tabulation cells. RESULTS: The main results are as follows: (1) In the support network of workplace associates, moderate drinkers enjoyed much more sociable support care than both nondrinkers and hard drinkers, which might suggest a similar effect as the French paradox. Meanwhile in the familial and kinship network, the more intervening care support was provided, the harder respondents' drinking practice. (2) The drinking network among Japanese people for both sexes is likely to be convergent upon certain types of network categories and not decentralized in various categories. This might reflect of the drinking culture of Japan, which permits people to drink everyday as a practice, especially male drinkers. Subsequently, solitary drinking is not optional for female drinkers. (3) Intervening network analysis showed that the harder the respondents' drinking practices, the more frequently their drinking behaviors were checked in almost all the categories of network. A rather complicated gender double-standard was found in the network of hard drinkers with their friends, particularly for female drinkers. Medical professionals played a similar intervening role for men as family and kinship networks but to a less degree than friends for females. CONCLUSION: The social network is considerably associated with respondents' drinking, providing both sociability for moderate drinkers and intervention for hard drinkers, depending on network categories. To minimize the risk of hard drinking and advance self-healthy drinking there should be more research development on drinking practice and the social network. PMID- 16316073 TI - [Nationwide survey of alcohol drinking and alcoholism among Japanese adults]. AB - PURPOSE: To investigate the characteristics of alcohol use among Japanese adults and prevalence of alcohol dependence in Japan, we conducted a nationwide survey on alcohol drinking behavior and alcohol dependence among Japanese adults using a representative sampling method. METHODS: We sampled 3500 adults from throughout the entire country using a stratified random sampling method with two-step stratification, and carried out a home visit interview survey. A total of 2547 people (72.8%) responded to the survey. The survey period was June, 2003. The questionnaire contained questions about the frequency and quantity of alcohol use, 'hazardous use of alcohol' and 'alcohol dependence' according to the ICD-10 definition, several screening scales on problem use of alcohol (CAGE, KAST, AUDIT), life-time prevalence of 24 alcohol related diseases, smoking status, dysgryphia, and nightcap drinking. RESULTS: The number of respondents was, 1184 males, and 1363 females. Lifetime alcohol drinking, and weekly drinking, and daily drinking rates were 95.1%, 64.4%, and 36.2% for males, 79.0%, 27.5%, and 7.5% for females, respectively. Average daily alcohol consumption was 3.7 units for males, and 2.0 units for females (1 unit = 10 g pure alcohol). The proportion of drinkers who drank alcohol 4 units or more daily was 28.9% for males, and 7.6% for females, and that for 6 units or more was 12.7% for males, and 3.4% for females. The proportion of flasher was 41.2% for males, and 35.0% for females. Among screening questions, problem drinking was most frequently identified using AUDIT (score 12 points or more, 150 persons), followed by KAST (2 points or more, 100 persons) and CAGE (2 points or more, 98 persons). The number of subjects who met the ICD-10 criteria for alcohol dependence was 24, while the number who engaged in hazardous alcohol use was 64. CONCLUSIONS: This study revealed that problem drinking and alcohol dependence are a serious problem in Japanese general population. The problem of females drinking may be growing. The government should emphasize the prevention of alcohol drinking problems in adults and continue the conduct of nationwide prevalence surveys to monitor the problem. PMID- 16316074 TI - Symptomatological structure of volatile solvent-induced psychosis: is "solvent psychosis" a discernible syndrome? AB - Solvent-induced psychosis has been clinically identified among patients suffering from dependence on volatile solvents and those in psychotic state due to chronic solvent use. To clarify the symptomatological difference between solvent-induced psychosis and schizophrenia, the principal component analysis with VARIMAX rotation was applied to the point and duration estimates of symptoms observed among the solvent group and among the schizophrenic group. There were no significant group differences in age and family history of any psychosis. The study findings are as follows: (1) It is difficult to distinguish two groups based on the prevalence rates of symptoms alone. (2) However, the principal component VARIMAX rotation analysis of the prevalence and duration observing among the solvent group revealed seven factors consisting of "amotivation", "intoxication", "emotional instability", "delusion", "hallucination", "disinhibition" and "memory". The seven factors explained 75.4% of the variance of the symptoms in this group. (3) The same analysis applied to the data from the schizophrenic group showed six factors consisting of "thought progression", "emotional instability", "amotivation (or negative symptoms)", "delusion", "hallucination" and "anxiety". These factors explained 62.9% of the variance in the data of the schizophrenic group. These results support clinical observations the "amotivational syndrome" may be a characteristic feature of patients suffering from solvent-induced psychosis. The results also suggest "solvent psychosis" is a discernible syndrome, and is distinctive from psychotic symptoms of typical schizophrenia. PMID- 16316075 TI - [Progress in diagnosis of viral infectious diseases: impact of rapid diagnosis kits]. PMID- 16316076 TI - [A case of metabolic syndrome with coronary artery disease advanced over 13 years. Case report]. AB - A 61-year-old man was diagnosed with obesity, diabetes mellitus, and hyperlipidemia associated with insulin resistance in 1988. His condition was complicated with asymptomatic coronary artery disease in 1992. His coronary artery disease gradually progressed during the subsequent 13 years of observation, and he underwent percutaneous coronary intervention four times and also received a coronary artery bypass graft. This is a case of metabolic syndrome with multiple risk factors for arteriosclerosis as visceral obesity, insulin resistance, diabetes, hypertension and hyperlipidemia, in which the recent rapid progression of coronary artery disease might be associated with the discontinuation of statin after coronary artery bypass graft, accompanied with hyper-LDL-choleterolemia. Patients with metabolic syndrome require most comprehensive and strict therapies against multiple risk factors. PMID- 16316077 TI - Self-assembly of patchy particles into diamond structures through molecular mimicry. AB - Fabrication of diamond structures by self-assembly is a fundamental challenge in making three-dimensional photonic crystals. We simulate a system of model hard particles with attractive patches and show that they can self-assemble into a diamond structure from an initially disordered state. We quantify the extent to which the formation of the diamond structure can be facilitated by "seeding" the system with small diamond crystallites or by introducing a rotation interaction to mimic a carbon-carbon antibonding interaction. Our results suggest patchy particles may serve as colloidal "atoms" and "molecules" for the bottom-up self assembly of three-dimensional crystals. PMID- 16316078 TI - Evolution of surface morphologies in multivariant assemblies of surface-tethered diblock copolymers after selective solvent treatment. AB - We study systematically the topography behavior of PHEMA-b-PMMA block as a function of the PHEMA and PMMA block lengths after selectively collapsing the top (PMMA) block by using surface-anchored assemblies of poly(2-hydroxyethyl methacrylate-b-methyl methacrylate), PHEMA-b-PMMA, block copolymer with orthogonally varying lengths of each block. Our experimental results are in excellent qualitative agreement with topology diagrams predicted by self consistent field calculations of Zhulina and co-workers. PMID- 16316079 TI - Formation of cubic phases from large unilamellar vesicles of dioleoylphosphatidylglycerol/monoolein membranes induced by low concentrations of Ca2+. AB - We developed a new method for the transformation of large unilamellar vesicles (LUVs) into the cubic phase. We found that the addition of low concentrations of Ca(2+) to suspensions of multilamellar vesicles (MLVs) of membranes of monoolein (MO) and dioleoylphosphatidylglycerol (DOPG) mixtures (DOPG/MO) changed their L(alpha) phase to the cubic phases. For instance, the addition of 15-25 mM Ca(2+) to 30%-DOPG/70%-MO-MLVs induced the Q(229) phase, whereas the addition of > or =28 mM Ca(2+) induced the Q(224) phase. LUVs of DOPG/MO membranes containing > or =25 mol % DOPG were prepared easily. Low concentrations of Ca(2+) transformed these LUVs in excess buffer into the Q(224) or the Q(229) phase, depending on the Ca(2+) concentration. For example, 15 and 50 mM Ca(2+) induced the Q(224) and Q(229) phase in the 30%-DOPG/70%-MO-LUVs at 25 degrees C, respectively. This finding is the first demonstration of transformation of LUVs of lipid membranes into the cubic phase under excess water condition. PMID- 16316080 TI - Growth of gold nanoparticles in human cells. AB - Gold nanoparticles of 20-100 nm diameter were synthesized within HEK-293 (human embryonic kidney), HeLa (human cervical cancer), SiHa (human cervical cancer), and SKNSH (human neuroblastoma) cells. Incubation of 1 mM tetrachloroaurate solution, prepared in phosphate buffered saline (PBS), pH 7.4, with human cells grown to approximately 80% confluency yielded systematic growth of nanoparticles over a period of 96 h. The cells, stained due to nanoparticle growth, were adherent to the bottom of the wells of the tissue culture plates, with their morphology preserved, indicating that the cell membrane was intact. Transmission electron microscopy of ultrathin sections showed the presence of nanoparticles within the cytoplasm and in the nucleus, the latter being much smaller in dimension. Scanning near field microscopic images confirmed the growth of large particles within the cytoplasm. Normal cells gave UV-visible signatures of higher intensity than the cancer cells. Differences in the cellular metabolism of cancer and noncancer cells were manifested, presumably in their ability to carry out the reduction process. PMID- 16316081 TI - Monitoring of the self-assembled monolayer of 1-hexadecanethiol on a gold surface at nanomolar concentration using a piezo-excited millimeter-sized cantilever sensor. AB - In this paper, we describe a new method of measuring alkanethiol monolayer formation on a gold surface. A gold-coated millimeter-sized rectangular-shaped lead zirconate titanate (PZT) cantilever of dimensions 3.5 x 2 x 0.05 mm, previously shown to detect a picogram level of mass change, was used to measure the adsorption kinetics of 1-hexadecanethiol in ethanol over six orders of concentration range (1 nM to 10 mM) in real time. The flexural mode of cantilever vibration, 45.5 +/- 0.01 kHz, was monitored during the self-assembly. The total resonant frequency change obtained for the 1 nM, 10 nM, 100 nM, 1 microM, 4 mM, 8 mM, and 10 mM thiol concentrations were 116 +/- 2 (n = 2), 225 (n = 1), 270 +/- 10 (n = 2), 440 +/- 10 (n = 2), 900 +/- 10 (n = 2), 900 +/- 10 (n = 2), and 900 +/- 10 (n = 2) Hz, respectively. These results compare favorably to literature results in that the rate of the monolayer formation is concentration-dependent and the exponential change during adsorption follows the reversible first-order Langmuir kinetic model. The rate constants of adsorption and desorption were 0.061 M(-1) s(-1) and 3.61 x 10(-4) s(-1), respectively. The significance of the results is that millimeter-sized PZT cantilevers can be used in real-time for characterizing self-assembly of monolayer formation at nanomolar concentration levels. In addition, at 1 nM, the adsorption was found not to be diffusion limited. PMID- 16316082 TI - Self-assembly and oligomerization of alkyne-terminated molecules on metal and oxide surfaces. AB - We report the self-assembly and subsequent oligomerization of organic molecules based on terthiophenes bearing a terminal alkyne moiety. Molecules with thioacetate and phosphonic acid functional groups were synthesized, enabling molecular self-assembly on metal (Au and Pd) and metal oxide [Al(2)O(3), HfO(2), and indium tin oxide (ITO)] surfaces, respectively. The molecules were assembled from solution and then oligomerized using either 2,5-norbornadiene-rhodium(I) chloride dimer or UV light. UV-vis and infrared absorption spectroscopies and electrochemical techniques show that the molecules assemble to form dense monolayers on the substrate surfaces and oligomerize under the action of a catalyst or UV light. PMID- 16316083 TI - Effects of compression and shearing on the microstructure of polymer-immobilized non-close-packed colloidal crystalline arrays. AB - We have examined the changes in the optical properties and microstructure of polymer-immobilized non-close-packed colloidal crystalline arrays with compression and shearing stress. The optical properties and microstructures of the arrays were measured by angle-resolved reflection spectroscopy. The spectra indicate an increase in the refractive index and a decrease in the interplane spacing with compression, however, indicating an increase in the interplane spacing with shearing stress. These results show that compression decreases the interplane spacing without moving the inner-plane position, while shearing stress increases the interplane spacing by moving the position of the spheres in the same plane. PMID- 16316084 TI - Multicomponent cationic lipid-DNA complex formation: role of lipid mixing. AB - Multicomponent cationic lipid-DNA complexes (lipoplexes) were prepared by adding linear DNA to mixed lipid dispersions containing two populations of binary cationic liposomes and characterized by means of small angle X-ray scattering (SAXS). Four kinds of cationic liposomes were used. The first binary lipid mixture was made of the cationic lipid (3'[N-(N',N'-dimethylaminoethane) carbamoyl]cholesterol (DC-Chol) and the neutral helper lipid dioleoylphosphocholine (DOPC) (DC-Chol/DOPC liposomes), the second one of the cationic 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and the neutral dioleoylphosphatidylethanolamine (DOPE) (DOTAP/DOPE liposomes), the third one of DC-Chol and DOPE (DC-Chol/DOPE liposomes), and the fourth one of DOTAP and DOPC (DOTAP/DOPC liposomes). Upon DNA-induced fusion of liposomes, large lipid mixing at the molecular level occurs. As a result, highly organized mixed lipoplexes spontaneously form with membrane properties intermediate between those of starting liposomes. By varying the composition of lipid dispersions, different DNA packing density regimes can also be achieved. Furthermore, occurring lipid mixing was found to induce hexagonal to lamellar phase transition in DOTAP/DOPE membranes. Molecular mechanisms underlying experimental findings are discussed. PMID- 16316085 TI - Formation of spatially patterned colloidal photonic crystals through the control of capillary forces and template recognition. AB - We report the formation of microscopic patterns of substrate-supported, 3D planar colloidal crystals using physical confinement in conjunction with surfaces displaying predetermined binary patterns of hydropholicity. The formation process involves a primary self-assembly wherein nano- and microscale colloids order into a photonic fcc lattice via capillary interactions followed by a secondary template-induced crystal cleavage step. Following this method, arbitrary arrays of pattern elements, which preserve structural and orientational properties of the parent crystal, can be easily obtained. PMID- 16316086 TI - Facile preparation of nanoelectrode ensembles using amphiphilic block copolymer film. AB - Taking advantage of both the self-organizing characteristics and the amphiphilic property of poly(styrene-block-ethyleneoxide) (P(S-b-EO)), we have realized one step fabrication of nanoelectrode ensembles (NEEs). By choosing the electrolyte solution elaborately, only the poly(ethylene oxide) (PEO) segment in the P(S-b EO) film was swollen to serve as nanoscale tunnels for solvated ions, whereas the polystyrene segment remained robust to be the electrode mask. An electrochemical analysis indicated that a transition between linear diffusion and nonlinear diffusion could be observed due to the small diffusion coefficient of ferrocene in PEO nanodomains. PMID- 16316087 TI - Width of the hexadecane-water interface: a discrepancy resolved. AB - An existing discrepancy in the determined values of the width of the hexadecane water interface as determined from X-ray reflection (XR) and recent neutron reflection (NR) measurements has been resolved. The NR-determined width reported here is 6.0 +/- 1.0 Angstroms, which compares extremely well with the XR determined value of 6.0 +/- 0.2 Angstroms. The origin of the discrepancy has been attributed to the thermal history of the interface rather than sample purity or other technique-dependent reasons. PMID- 16316088 TI - First principles computational study for understanding the interactions between ssdna and gold nanoparticles: adsorption of methylamine on gold nanoparticulate surfaces. AB - We conducted a computational adsorption study of methylamine on various surface models of a gold nanoparticle which is facetted by multiple [111] and [100] planes. In addition to these flat surfaces, our models include the stepped surfaces (ridges) formed along the intersections of these planes. Binding on the flat surfaces was fairly weak, but substantially stronger on the ridges by an average of 4.4 kcal/mol. This finding supports the idea that ssDNA's interaction with gold nanoparticles occurs through the amines on the purine/pyrimidine rings. Also, this typically undesirable interaction between DNA and gold nanoparticles is expected to increase as the particle size decreases. Our analysis suggests that particle size is an important controlling parameter to reduce this interaction. PMID- 16316089 TI - Poly[bis(2,2,2-trifluoroethoxy)phosphazene] superhydrophobic nanofibers. AB - Nanofibers of poly[bis(2,2,2-trifluoroethoxy)phosphazene] were produced by electrospinning from solutions in tetrahydrofuran, methylethyl ketone, and acetone. The fiber diameter varied from 80 nm to 1.4 microm by changes in the concentration of the polymer solution. The electrospun nonwoven mats showed enhanced surface hydrophobicity compared to spun cast films with up to a 55 degrees increase in water contact angle. The hydrophobicity varied with fiber diameter and surface morphology, with contact angles to water being in the range of 135 degrees -159 degrees. A low value of hysteresis (<4 degrees) was recorded for the superhydrophobic surfaces. The extremely high hydrophobicity of these mats is a combined result of a fluorinated surface and the inherent surface roughness of an electrospun mat. PMID- 16316090 TI - Stable dispersions of silver nanoparticles in carbon dioxide with fluorine-free ligands. AB - Iso-stearic acid, a short, stubby compound with branched, methylated tails has been shown to have high solubility in carbon dioxide. Tail solvation by carbon dioxide makes iso-stearic acid a good choice for use as a ligand to sterically stabilize metallic nanoparticles. Iso-stearic acid coated silver nanoparticles have been stably dispersed in carbon dioxide with hexane cosolvent. Neat carbon dioxide has successfully dispersed iso-stearic acid coated silver nanoparticles that had been deposited on either quartz or polystyrene surfaces. These results are the first reports of sterically stabilized nanoparticles in carbon dioxide without the use of any fluorinated compounds. PMID- 16316091 TI - Continuous microfluidic reactors for polymer particles. AB - This article provides an overview of our work in the area of the synthesis of polymer particles in continuous microfluidic reactors. The method includes (a) the generation of highly monodisperse monomer droplets in a microfluidic flow focusing device and (b) in-situ solidification of these droplets by means of photopolymerization. We discuss the effect of monomer properties on the emulsification process, the effect of the polymerization rate on the production of high-quality particles, the role of the material of the microfluidic device in droplet formation, and the synthesis of particles with different shapes and compositions. We also demonstrate the production of highly ordered arrays of polymer particles achieved by photopolymerization of the dynamic lattices of monomer droplets in microfluidic channels. The article is concluded with a summary of future research directions in the production of polymer colloids in microfluidic reactors. PMID- 16316092 TI - Frequency-dependent shape changes of colloidal clusters under transverse electric field. AB - We have studied clustering of colloidal particles under the influence of an ac electric field as a function of frequency. The field was applied in a direction perpendicular to the confining walls. Two regimes are observed, a low frequency regime where the clusters are isotropic with a local triangular order, as reported earlier in the literature, and a new high-frequency regime where the clusters are highly elongated (anisotropic) with no local order. The crossover from one regime to the other occurs at a critical frequency, f(c). The threshold field for the cluster formation, E(th), increases with frequency in both the regimes. An increase in the particle size leads to a reduction in both E(th) and f(c). We present evidence to show that the elongated structures seen at high frequency are related to the field inhomogeneities at imperfections on the conducting surface. We also propose a possible mechanism based on hydrodynamic flow considerations to explain the formation of these clusters. PMID- 16316093 TI - Tetrabutylammonium alkyl carboxylate surfactants in aqueous solution: self association behavior, solution nanostructure, and comparison with tetrabutylammonium alkyl sulfate surfactants. AB - A series of long and ultralong chain tetrabutylammonium alkyl carboxylate (TBACm, TBA = tetrabutylammonium ion; Cm = carboxylate ion C(m-1)H(2)(m-1)CO(2)(-) of total carbon number m) surfactants have been obtained by direct neutralization of the fatty acids with m = 12, 14, 18, 22, and 24 by tetrabutylammonium hydroxide. Time-resolved fluorescence quenching has been used to determine the micelle aggregation number (N) of the surfactants with m = 12, 14, and 18 in the temperature range 10-50 degrees C and of the surfactants with m = 22 and 24 in the temperature range 25-60 degrees C. In all instances the values of N were well below those that can be calculated for the maximum spherical micelle formed by surfactants with the same alkyl chain as the investigated surfactants on the basis of the oil drop model for the micelle core. The microstructure of selected solutions of TBAC22 was examined using transmission electron microscopy at cryogenic temperature and compared to the microstructure of solutions of TBA dodecyl and tetradecyl sulfates. These observations generally confirmed the findings of TRFQ. The self-association behavior of these anionic surfactants with TBA counterions is explained on the basis of the large size and the hydrophobicity of the tetrabutylammonium ions. The important differences in behavior that have been evidenced between tetrabutylammonium alkyl carboxylates and alkyl sulfates are discussed in terms of differences in distribution of the surfactant electrical charge on the headgroup and alkyl chain predicted by quantum chemical calculations (Langmuir 1999, 15, 7546). PMID- 16316094 TI - Self-diffusion NMR studies of the host-guest interaction between beta cyclodextrin and alkyltrimethylammonium bromide surfactants. AB - Diffusion measurements by nuclear magnetic resonance (NMR) spectroscopy were used to investigate the host-guest association between beta-cyclodextrin (CD) and alkyltrimethylammonium bromide surfactants with different chain lengths, ranging from 6 up to 16 carbons. The scope and limitations of the method in the study of formation of inclusion complexes are discussed. The influences of the presence of CD in the micellization process have been studied, and the apparent critical micellar concentration and the self-diffusion coefficients of the species present in the systems have been calculated. The stoichiometries of the different complexes have been determined. Evidence for the formation of a 2:1 complex in the case of C(16)TAB has been found. PMID- 16316095 TI - A new method for obtaining adsorption isotherms on colloidal suspensions via electrokinetic sonic amplitude measurement. AB - The standard methods for obtaining adsorption isotherms on colloidal suspensions are usually very time consuming and involve a large number of steps and assumptions that increase the experimental errors. In this work, an alternative method is proposed to evaluate the adsorption behavior of electrosteric stabilized systems based on electrokinetic sonic amplitude signal measurements. The new method, entitled "zeta-sorption", is noticeably less time-consuming when compared to conventional procedures but showed great precision and reliability confirmed by comparison with data obtained from conventional routes on alumina polyacrylate and alumina-citric acid aqueous suspensions. The experimental conditions that restrict the applicability of the new method were identified and justified by discussing the possible ion exchanges. PMID- 16316096 TI - Rheology of asphaltene-toluene/water interfaces. AB - The stability of water-in-crude oil emulsions is frequently attributed to a rigid asphaltene film at the water/oil interface. The rheological properties of these films and their relationship to emulsion stability are ill defined. In this study, the interfacial tension, elastic modulus, and viscous modulus were measured using a drop shape analyzer for model oils consisting of asphaltenes dissolved in toluene for concentrations varying from 0.002 to 20 kg/m(3). The effects of oscillation frequency, asphaltene concentration, and interface aging time were examined. The films exhibited viscoelastic behavior. The total modulus increased as the interface aged at all asphaltene concentrations. An attempt was made to model the rheology for the full range of asphaltene concentration. The instantaneous elasticity was modeled with a surface equation of state (SEOS), and the elastic and viscous moduli, with the Lucassen-van den Tempel (LVDT) model. It was found that only the early-time data could be modeled using the SEOS-LVDT approach; that is, the instantaneous, elastic, and viscous moduli of interfaces aged for at most 10 minutes. At longer interface aging times, the SEOS-LVDT approach was invalid, likely because of irreversible adsorption of asphaltenes on the interface and the formation of a network structure. PMID- 16316097 TI - Transient electrophoresis of spherical particles at low potential and arbitrary double-layer thickness. AB - A theoretical study is presented for the dynamic electrophoretic response of a charged spherical particle in an unbounded electrolyte solution to a step change in the applied electric field. The electric double layer surrounding the particle may have an arbitrary thickness relative to the particle radius. The transient Stokes equations modified with the electrostatic effect which govern the fluid velocity field are linearized by assuming that the system is only slightly distorted from equilibrium. Semianalytical results for the transient electrophoretic mobility of the particle are obtained as a function of relevant parameters by using the Debye-Huckel approximation. The results demonstrate that the electrophoretic mobility of a particle with a constant relative mass density at a specified dimensionless time normalized by its steady-state quantity decreases monotonically with a decrease in the parameter kappaa, where kappa(-1) is the Debye screening length and a is the particle radius. For a given value of kappaa, a heavier particle lags behind a lighter one in the development of the electrophoretic mobility. In the limits of kappaa --> infinity and kappaa = 0, our results reduce to the corresponding analytical solutions available in the literature. The electrophoretic acceleration of the particle is a monotonic decreasing function of the time for any fixed value of kappaa. In practical applications, the effect of the relaxation time for the transient electrophoresis is negligible, regardless of the value of kappaa or the relative mass density of the particle. PMID- 16316098 TI - Tethered bilayer lipid membranes based on monolayers of thiolipids mixed with a complementary dilution molecule. 1. Incorporation of channel peptides. AB - Tethered bilayer lipid membranes (tBLMs) are described based on the self-assembly of a monolayer on template stripped gold of an archea analogue thiolipid, 2,3-di o-phytanyl-sn-glycerol-1-tetraethylene glycol-d,l-alpha-lipoic acid ester lipid (DPTL), and a newly designed dilution molecule, tetraethylene glycol-d,l-alpha lipoic acid ester (TEGL). The tBLM is completed by fusion of liposomes made from a mixture of diphytanoylphosphatidyl choline (DPhyPC), cholesterol, and 1,2 diphytanoyl-sn-glycero-3-phosphate (DPhyPG) in a molar ratio of 6:3:1. Melittin and gramicidin are incorporated into these tBLMs as shown by surface plasmon resonance (SPR) and electrochemical impedance spectroscopy (EIS) studies. Ionic conductivity at 0 V vs Ag|AgCl, 3 M KCl, measured by EIS measurements are comparable to the results obtained by other research groups. Admittance plots as a function of potential are discussed on a qualitative basis in terms of the kinetics of ion transport through the channels. PMID- 16316099 TI - pH-dependence of the properties of hydrophobically modified polyvinylamine. AB - A series of N-alkyl or N-benzyl substituted polyvinylamines (PVAm) were prepared and the properties of aqueous solutions were measured as functions of pH. The polymer solutions showed almost no surface activity under acidic conditions whereas surface tension was reduced to 40-50 mN/m around pH 9. Increasing either the degree of hydrophobic substitution or the hydrophobic chain length lowered the pH at which surface tension lowering was observed. Hydrophobic substitution also shifted plots of the degree of ionization versus pH toward lower pH which means lower pH values were required to achieve a given value of polymer charging. The hydrophobically modified PVAm associated in water giving species whose apparent diameter measured by dynamic light scattering decreased with increasing pH, whereas the electrophoretic mobilities of the associated species increased with decreasing pH. Although many hydrophobically modified and pH sensitive polymers have been described in the literature for applications in biomaterials, drug release and as pH sensitive surfactants, the hydrophobically modified PVAms are particularly attractive because they are easily prepared from commercially available polyvinylamines. PMID- 16316100 TI - Gas-solid coexistence in highly charged colloidal suspensions. AB - Aqueous suspensions of highly charged polystyrene particles with different volume fractions have been investigated for structural ordering and phase behavior using static light scattering (SLS) and confocal laser scanning microscope (CLSM). Under deionized conditions, suspensions of high-charge-density colloidal particles remained disordered whereas suspensions of relatively low charge density showed crystallization by exhibiting iridescence for the visible light. Though for the unaided eye crystallized suspensions appeared homogeneous, SLS measurements and CLSM observations have revealed their inhomogeneous nature in the form of the coexistence of voids with dense ordered regions. CLSM investigations on disordered suspensions showed their inhomogeneous nature in the form coexistence of voids with dense disordered (amorphous) regions. Our studies on highly charged colloids confirm the occurrence of gas-solid transition and are in accordance with predictions of Monte Carlo simulations using a pair-potential having a long-range attractive term [Mohanty, P. S.; Tata, B. V. R. J. Colloid Interface Sci. 2003, 264, 101]. On the basis of our experimental and simulation results, we argue that the reported reentrant disordered state [Yamanaka et al. Phys. Rev. Lett. 1998, 80, 5806 and Toyotama et al. Langmuir 2003, 19, 3236] in charged colloids observed at high charge densities is a gas-solid coexistence state. PMID- 16316101 TI - Direct observation of thiolate displacement reactions on Au(111): the role of physisorbed disulfides. AB - Line-of-sight mass spectroscopy (LOSMS) has been used to study the displacement reaction of ( radical3x radical3)R30 degrees methylthiolate on Au(111) by butylthiolate. The reaction was carried out at room temperature and constant saturation coverage, by exposing the methylthiolate-covered surface to dibutyl disulfide gas. The adsorbed methylthiolate was desorbed as dimethyl disulfide and the cross product methylbutyl disulfide, both identified by LOSMS. This shows that reaction occurs between adsorbed thiolates of different chain length at room temperature, while the kinetics indicate that a rapid equilibrium is established between immobile, chemisorbed thiolates, and highly mobile, physisorbed disulfides. PMID- 16316102 TI - Inhibition of aluminum oxyhydroxide precipitation with citric acid. AB - Citric acid has been shown to act as an agent for increasing the solubility of aluminum oxyhydroxides in aqueous solutions of high (>2.47 mol/mol) hydroxide-to aluminum ratios. Conversely, citric acid also colloidally stabilizes particles in aqueous suspensions of aluminum-containing particles. Solutions of aluminum chloride, with and without citric acid added, were titrated with NaOH(aq). The presence and size of particles were determined using quasi-elastic light scattering. In solutions that contained no citric acid, particles formed instantaneously when NaOH(aq) was added but these were observed to rapidly diminish in size, disappearing at OH/Al ratios below 2.5 mol/mol. When the OH/Al ratio was raised beyond 2.5 by adding more NaOH(aq), suspensions of colloidally stable particles formed. Large polycations containing 13 aluminum atoms were detected by (27)Al solution NMR in citric-acid-free solutions with OH/Al ratios slightly lower than 2.5. In comparison, adding citric acid to solutions of aluminum chloride inhibited the formation of large aluminum-containing polycations. The absence of the polycations prevents or retards the subsequent formation of particles, indicating that the polycations, when present, act as seeds to the formation of new particles. Particles did not form in solutions with a citric acid/aluminum ratio of 0.8 until sufficient NaOH(aq) was added to raise the OH/Al ratio to 3.29. By comparison, lower amounts of citric acid did not prevent particles from forming but did retard the rate of growth. PMID- 16316103 TI - Polymer surfaces with reversibly switchable ordered morphology. AB - Honeycomb macroporous films fabricated by the "breath figures" method were composed of poly2-vinylpyridine (P2VP) distributed in the holes of polystyrene (PS). The porous films exhibited reversible behavior responding to water and different solvent vapors. When the porous film was treated with water, the honeycomb pattern would change to the hexagonal islandlike pattern. Once heated to remove the water, the honeycomb pattern emerged again. When the porous film was exposed to different solvent vapors, the same reversible process appeared. Carbon disulfide (CS(2)), toluene (TOL), and tetrahydrofuran (THF) solvent vapors induced the honeycomb pattern into the ordered islandlike pattern, and ethanol, chloroform, methyl ethyl ketone (MEK), and dimethylformamide (DMF) solvent vapors made the islandlike pattern come back to the honeycomb pattern. The hygroscopic property of P2VP and the polymer-solvent interaction are the driving force for the reversibly switchable morphology. The appropriate control of the hole depth is very crucial in determining the reversible changes. PMID- 16316104 TI - Neutron reflection from the liquid-liquid interface: adsorption of hexadecylphosphorylcholine to the hexadecane-aqueous solution interface. AB - Adsorption of water-soluble, zwitterionic n-hexadecylphosphorylcholine (C(16)PC) amphiphiles has been examined at the hexadecane-aqueous solution interface using neutron reflectivity (NR) and interfacial tension measurements. The results of both methods indicate that the limiting area per surfactant molecule at the interface at the critical micelle concentration (cmc) is 40 +/- 5 Angstroms(2). In the NR measurements, two isotopic contrasts have been employed to determine the adsorption isotherm and to explore the structure of the interfacial region. Single-layer model fitting to both isotopic contrasts was only possible for the single sub-cmc concentration studied, where a film thickness of 60 +/- 5 Angstroms was obtained; consistent single-layer model fits to both contrasts for concentrations greater than the cmc were not possible, leading to the requirement of a two-layer model with an overall film thickness close to 60 +/- 2 Angstroms. This film thickness is appreciably greater than the fully extended C(16)PC molecular length and cannot be explained purely in terms of thermal broadening. A further result is that the reflectivity data indicate that, as the C(16)PC concentration increases, the amount of water on the hexadecane side of the interfacial region increases, in contrast to intuitive expectation. These findings are interpreted by conjecturing a structural model in which a trilayer of C(16)PC molecules is formed at the interface with the water concentrated in the region occupied by the headgroups. PMID- 16316105 TI - Interactions between a solid spherical particle and a chemically heterogeneous planar substrate. AB - The Derjaguin-Landau-Verwey-Overbeek (DLVO) interaction forces between a chemically heterogeneous substrate and a spherical particle approaching the surface are calculated using a simple model. This model decomposes the heterogeneous substrate into nanoscale subunits and applies a pairwise summation of forces between each subunit and the approaching particle to determine the net DLVO force. This approach leads to a three-dimensional description of the forces arising from the chemical heterogeneity of the surface. In this three-dimensional realm, we observe the emergence of a substantial lateral force at specific separation distances from the substrate. More specifically, the lateral forces become significantly larger than the normal forces at separation distances between those of the repulsive barrier and the secondary minimum of the DLVO interaction energy curve. These lateral forces are most pronounced at high electrolyte concentrations, particularly at biological salt concentrations of approximately 0.1 M. Furthermore, the lateral forces are found to be significantly higher when the particle is near the edge of a heterogeneous region of the substrate. On the basis of the evidence of this study, and depending on the characteristics of the system, both the physical roughness and the chemical heterogeneity of a surface can significantly affect how a particle will interact with it. PMID- 16316106 TI - A [4+2]-like cycloaddition of methyl methacrylate on Si(100)-2 x 1. AB - The attachment of methyl methacrylate (MMA) on Si(100)-2x1 was investigated using high-resolution electron energy loss spectroscopy (HREELS), X-ray photoelectron spectroscopy (XPS), ultraviolet photoelectron spectroscopy (UPS), and density functional theory (DFT) calculations. The HREELS spectra of chemisorbed MMA show the disappearance of characteristic vibrations of C=O (1725 cm(-1)) and C(sp(2)) H (3110, 1400, and 962 cm(-1)) coupled with the blue shift of the C=C stretching mode by 34 cm(-1) compared to those of physisorbed molecules. These results clearly demonstrate that both C=C and C=O in MMA directly participate in the interaction with the surface to form a SiCH(2)C(CH(3))=C(OCH(3))OSi species via a [4+2]-like cycloaddition. This binding configuration was further supported by XPS, UPS, and DFT studies. PMID- 16316107 TI - Hydrophobization of glass surface by adsorption of poly(dimethylsiloxane). AB - Silica or glass particles are introduced in a poly(dimethylsiloxane) (PDMS) matrix for various applications. A particular feature of these systems is that PDMS adsorbs on the surface of the dispersed particles, thus rendering them more hydrophobic with time. The mechanism of this process of in situ hydrophobization is still poorly understood. The major aims of the present study are (1) to quantify the rate of surface hydrophobization by PDMS and, on this basis, to discuss the mechanism of the process; (2) to compare the contact angles of surfaces that are hydrophobized by different procedures and are placed in contact with different fluid interfaces-PDMS-water, hexadecane-water, and air-water; and (3) to check how the type of surfactant affects the contact angles, viz., the effective hydrophobicity of the surface. We present experimental results for the kinetics of hydrophobization of glass surfaces, which are characterized by measuring the three-phase contact angle of glass-surfactant solution-PDMS. The data reveal two consecutive stages in the hydrophobization process: The first stage is relatively fast and the contact angle increases from 0 degrees to about 90 degrees within several minutes. This stage is explained with the physical adsorption of the PDMS chains, as a result of hydrogen-bond formation with the surface silanol groups. The second stage is much slower and hours or days are required at room temperature to reach the final contact angle (typically, 150-160 degrees). This stage is explained as grafting of the PDMS molecules on the surface by chemical reaction with the surface silanol groups. If the glass surface had been pretreated by hexamethyldisilazane (HMDS), so that CH(3) groups had blocked most of the surface silanol groups, the first stage in the hydrophobization process is almost missing-the contact angle slowly changes at room temperature from about 90 degrees up to 120 degrees. The experiments aimed to compare several hydrophobization procedures showed that PDMS ensures larger contact angle (more hydrophobic surface) than grafted alkyl chains. The contact angles at the PDMS-water and hexadecane-water interfaces were found to be very similar to each other, and much larger than that at the air-water interface. Interestingly, we found that the ionic surfactants practically do not affect the contact angle of PDMS-hydrophobized surface, whereas the nonionic surfactants reduce this angle. Similar trends are expected with silica surfaces, as well. PMID- 16316108 TI - Controlling polymer adhesion with "pancakes". AB - Topographical patterns are used to selectively tune the adhesion of polymers. Although nature has provided guidance, relatively little is known of how topographic patterns can be intelligently used not only to enhance adhesion but, more importantly, to tune adhesion. We demonstrate that properly designed, low aspect-ratio posts can alter adhesion from 20% to 400% the value of conventional adhesion descriptors for nonpatterned interfaces. This control is not related to the magnitude of interfacial area but, rather, to altering the local separation processes at an interface by geometry. We establish general relationships that govern the interaction between material properties, pattern length scales, and the control of adhesion. These relationships provide insight into the mechanisms of adhesion for examples in nature, such as the gecko, while also providing concrete guidance for the future design of "smart" adhesives and coatings with nanoscale patterns. PMID- 16316109 TI - Tribological properties of alkylsilane self-assembled monolayers. AB - In this study, we perform molecular dynamics simulations of adhesive contact and friction between alkylsilane Si(OH)(3)(CX(2))(10)CX(3) and alkoxylsilane Si(OH)(2)(CX(2))(10)CX(3) (where X = H or F) self-assembled monolayers (SAMs) on an amorphous silica substrate. The alkylsilane SAMs are primarily hydrogen-bonded or physisorbed to the surface. The alkoxylsilane SAMs are covalently bonded or chemisorbed to the surface. Previously, we studied the chemisorbed systems. In this work, we study the physisorbed systems and compare the tribological properties with the chemisorbed systems. Furthermore, we examine how water at the interface of the SAMs and substrate affects the tribological properties of the physisorbed systems. When less than a third of a monolayer is present, very little difference in the microscopic friction coefficient mu or shear stresses is observed. For increasing amounts of water, the values of mu and the shear stresses decrease; this effect is somewhat more pronounced for fluorocarbon alkylsilane SAMs than for the hydrocarbon SAMs. The observed decrease in friction is a consequence of a slip plane that occurs in the water as the amount of water is increased. We studied the frictional behavior using relative shear velocities ranging from v = 2 cm/s to 2 m/s. Similar to previously reported results for alkoxylsilane SAMs, the values of the measured stress and mu for the alkylsilane SAM systems decrease monotonically with v. PMID- 16316110 TI - Polarization of water near dipolar surfaces: a simple model for anomalous dielectric behavior. AB - A model for the electrostatic interactions in water in the vicinity of a surface is suggested, which accounts, within the Poisson-Boltzmann mean field approach, for the screening of the charges and for the coupling interactions between neighboring dipoles. When the water molecules near a solid surface are assumed to be organized in icelike layers, the polarization is not a continuous function but exists only at the discrete positions of the water molecules. The particular positions of the water molecules in the icelike structure govern the manner in which the average water dipoles align with each other. On the basis of this model, one could explain the nonmonotonic behavior of the polarization and the electrical potential as well as the anomalous dielectric response of water (the nonproportionality of the polarization and the macroscopic electric field), which were obtained recently via molecular dynamics simulations. PMID- 16316111 TI - Polyelectrolyte modified solid surfaces: the consequences for ionic and mixed ionic/nonionic surfactant adsorption. AB - This paper describes how the cationic polyelectrolyte, polyDMDAAC (poly(dimethyl diallylammonium chloride)), is used to manipulate the adsorption of the anionic surfactant SDS and the mixed ionic/nonionic surfactant mixture of SDS (sodium dodecyl sulfate)/C(12)E(6) (monododecyl hexaethylene glycol) onto the surface of hydrophilic silica. The deposition of a thin robust polymer layer from a dilute polymer/surfactant solution promotes SDS adsorption and substantially modifies the adsorption of SDS/C(12)E(6) mixtures in favor of a surface relatively rich in SDS compared to the solution composition. Different deposition conditions for the polyDMDAAC layer are discussed. In particular, at higher solution polymer concentrations and in the presence of 1 M NaCl, a thicker polymer layer is deposited and the reversibility of the surfactant adsorption is significantly altered. PMID- 16316112 TI - Reactivity control of carboxylic acid-terminated self-assembled monolayers on gold: acid fluoride versus interchain carboxylic anhydride. AB - Reactions that occur at interfaces often show different behaviors from their solution analogues. In this paper, we demonstrated how proximity effect, one of the unique phenomena at interfaces, could control the product distributions of interfacial reactions. Self-assembled monolayers (SAMs) of 16 mercaptohexadecanoic acid on gold surfaces were treated with cyanuric fluoride and pyridine, which are generally used for forming acid fluorides from carboxylic acids in the solution-based reaction. After the treatment, two different products, acid fluorides (AFs) and interchain carboxylic anhydrides (ICAs), were controllably obtained at surfaces under different reaction conditions with keeping the reagents the same. Various factors, such as the concentrations of reagents, reaction time, and additives, affected the product distribution (or the reaction pathway) at surfaces. We found that one of the key factors in controlling the reaction pathway was a relative contribution from the proximity effect of adjacent carboxylic acid chains in the SAMs (kinetic control) and the equilibrium shift (thermodynamic control). The relative reactivity of AF- and ICA presenting surfaces toward primary amines, such as undecylamine and [((6 aminohexyl)amino)carbonyl]ferrocene, was also investigated, in terms of the number and the ordering of the amines coupled onto the surfaces. PMID- 16316113 TI - Photoinitiated chemical vapor deposition of polymeric thin films using a volatile photoinitiator. AB - Photoinitiated chemical vapor deposition (piCVD) is an evolutionary CVD technique for depositing polymeric thin films in one step without using any solvents. The technique requires no pre- or post-treatment and uses a volatile photoinitiator to initiate free-radical polymerization of gaseous monomers under UV irradiation. Glycidyl methacrylate (GMA) was used as a test monomer for its ability to undergo free-radical polymerization, and 2,2'-azobis(2-methylpropane) (ABMP) was used as the photoinitiator, as it is known to produce radicals when excited by photons. GMA and ABMP vapors were fed into a vacuum chamber in which film growth was observed on a substrate exposed to UV irradiation. The resulting poly(glycidyl methacrylate) (PGMA) thin films were comprised of linear chains and had high structural resemblance to conventionally polymerized PGMA, as shown by the high solubility in tetrahydrofuran and the infrared and X-ray photoelectron spectroscopy measurements. The introduction of ABMP into the vacuum chamber significantly increased growth rates. The maximum growth rate achieved was approximately 140 nm/min and represents a 7-fold enhancement over the case without ABMP. The molecular weight was found to increase with increasing monomer to-initiator (M/I) feed ratio, and the polydispersity indexes (PDIs) of the samples were between 1.8 and 2.2, lower than the values obtained in conventional batch polymerization but in agreement with the theoretical expressions developed for low-conversion solution-phase polymerization, which are applicable to continuous processes such as piCVD. Molecular-weight distributions can be narrowed by filtering out wavelengths shorter than 300 nm, which induce branching and/or cross-linking. The strong dependence of the molecular weight on the M/I ratio, the rate enhancement due to the use of a radical photoinitiator, the good agreement between the experimental, and the theoretical PDIs provide evidence of a free-radical mechanism in piCVD. The clear films obtained in this work had number-average molecular weights between 12 500 and 97 000 g/mol. The similarities in growth conditions, growth rates, and molecular weights between the initiated CVD, a previously reported thermal process able to synthesize a wide range of polymers, and the piCVD of PGMA suggest that piCVD can also be used to produce those polymers and potentially others whose monomers undergo free radical mechanisms. This paper serves as an introduction to the technique by demonstrating piCVD's ability in synthesizing high-molecular-weight PGMA thin films with narrow molecular-weight distributions from vapors of GMA and ABMP in a single, dry step under UV irradiation. PMID- 16316114 TI - Enhanced and selective adsorption of mercury ions on chitosan beads grafted with polyacrylamide via surface-initiated atom transfer radical polymerization. AB - Enhanced and selective removal of mercury ions was achieved with chitosan beads grafted with polyacrylamide (chitosan-g-polyacrylamide) via surface-initiated atom transfer radical polymerization (ATRP). The chitosan-g-polyacrylamide beads were found to have significantly greater adsorption capacities and faster adsorption kinetics for mercury ions than the chitosan beads. At pH 4 and with initial mercury concentrations of 10-200 mg/L, the chitosan-g-polyacrylamide beads can achieve a maximum adsorption capacity of up to 322.6 mg/g (in comparison with 181.8 mg/g for the chitosan beads) and displayed a short adsorption equilibrium time of less than 60 min (compared to more than 15 h for the chitosan beads). Coadsorption experiments with both mercury and lead ions showed that the chitosan-g-polyacrylamide beads had excellent selectivity in the adsorption of mercury ions over lead ions at pH < 6, in contrast to the chitosan beads, which did not show clear selectivity for either of the two metal species. Mechanism study suggested that the enhanced mercury adsorption was due to the many amide groups grafted onto the surfaces of the beads, and the selectivity in mercury adsorption can be attributed to the ability of mercury ions to form covalent bonds with the amide. It was found that adsorbed mercury ions on the chitosan-g-polyacrylamide beads can be effectively desorbed in a perchloric acid solution, and the regenerated beads can be reused almost without any loss of adsorption capacity. PMID- 16316115 TI - Relationship between interfacial transfer and adsorption-desorption of surface active bis-ammonium ions at a liquid/liquid interface. AB - Cyclic voltammograms and interfacial tension-applied potential curves were recorded at the interface between water containing surface-active bis-quaternary ammonium ions, bis-A(2+), and an organic solvent such as 1,2-dichloroethane or nitrobenzene. An ordinary diffusion-controlled voltammetric wave for the transfer of bis-A(2+) from aqueous phase to organic phase, the first wave, was followed by a typical adsorption-related wave, the second wave. It was found from the potential dependence of the interfacial tension of bis-A(2+) that the second wave was due to the desorption of bis-A(2+) toward the organic phase. The influence of the structure of bis-A(2+) on voltammograms was investigated, and the potential for the first wave was found to depend on both the length of the side chain and that of the spacer chain, whereas the potential for the second wave depended on the latter only. The thermodynamic relations among three processes of the ion transfer, adsorption, and desorption were discussed based on the experimental results. PMID- 16316116 TI - Surface hydration and its effect on fluorinated SAM formation on SiO2 surfaces. AB - Substrate hydration is demonstrated to be crucial to film quality during self assembled (SA) film deposition of tridecafluoro-1,1,2,2, tetrahydrooctyltrichlorosilane (FOTS) from the vapor phase. The surface hydration was studied by thermogravimetric analysis, and a model was developed to predict the conditions necessary to desorb all of the water adsorbed on a fused silica surface without significantly altering the concentration of the surface hydroxyl groups. The nature of the SA film was investigated as a function of the degree of rehydration of the dehydrated silica surface. The wettability and microstructure of the SA films were examined by water contact angle, ellipsometry, X-ray photoelectron spectroscopy, and atomic force microscopy. There is an optimum degree of substrate hydration, on the order of 1-1.2 monolayers of adsorbed water, required to produce a dense, durable and uniform FOTS film with high water repellency and a smooth surface. PMID- 16316117 TI - Photochemical reaction of trimethyl acetate on Pt/TiO2(110). AB - Nanometer-sized Pt particles were prepared on an atomically flat surface of rutile TiO(2). Trimethyl acetate (TMA) adsorbed on the Pt-modified surface was photochemically decomposed under ultraviolet light irradiation in a vacuum. Residing TMA anions were imaged by a scanning tunneling microscope to deduce the local rate of decomposition. Increasing the number density of Pt particles led to an enhancement of the initial reaction rate. The degree of this enhancement did not depend on the distance from the Pt particles. PMID- 16316118 TI - Synthesis of high-density grafted polymer layers with thickness and grafting density gradients. AB - A novel approach was developed for the synthesis of tethered polymer layers with thickness and grafting density gradients. Poly(glycidyl methacrylate) (PGMA) was employed as a primary anchoring layer to attach the polymer chains to the surface of a silicon wafer. A linear temperature gradient heated stage was used for the generation of a gradual variation in the thickness of the anchoring PGMA film along the substrate. The obtained gradient was translated into the polymerization initiator gradient via the reaction between the epoxy groups of PGMA and the carboxyl functionality of 2-bromo-2-methylpropionic acid (BPA). The attachment of BPA to the surface modified with the monolayer of PGMA was confirmed by X-ray photoelectron spectroscopy experiments. To complete the experimental procedures, surface-initiated atom transfer radical polymerization was performed to synthesize the grafted polymer layers with thickness and surface densities that were varied along the substrate. The grafting density of the samples created in this three-step process ranged from 0.75 +/- 0.05 to 1.5 +/- 0.25 chains/nm(2). It was estimated, from a comparison of the surface densities of the initiator and the attached polymer, that the efficiency of the initiation from the surface was on the order of 5-10% and was dependent upon the surface concentration of the initiator and the time of polymerization. PMID- 16316119 TI - Surface enhanced vibrational (IR and Raman) spectroscopy in the design of chemosensors based on ester functionalized p-tert-butylcalix[4]arene hosts. AB - Surface-enhanced IR (SEIR) and Raman scattering (SERS) have been employed to study the adsorption of ester functionalized tert-butyl calix[4]arenes on Ag and Au nanostructured surfaces as well as their complexes with pyrene. The influence of adsorption and complexation with pyrene on the host calixarene structure was tested for two different calixarene molecules bearing carboethoxy groups (CH(3)CH(2)COOCH(2)-) in the low rim at positions 1,3- and 1,2,3,4-. The results obtained with SEIR were compared to those obtained with SERS, to better understand the interaction mechanism of the studied calixarenes with the metallic surfaces and the ligand as well as to investigate the structure/selectivity relationship of these two surface techniques in the analysis of recognition problems in which these ester functionalized calixarene molecules are involved. PMID- 16316120 TI - FTIR spectroscopy study of CO adsorption on Pt-Na-mordenite. AB - Different carbonyls are formed after CO adsorption at ambient temperature on a Pt Na-mordenite (Pt-Na-MOR) sample. Pt(3+)(CO)(2) dicarbonyls (nu(s) at 2205 cm(-1) and nu(as) at 2167 cm(-1)) are decomposed without formation of monocarbonyls. The respective mixed-ligand species, Pt(3+)((12)CO)((13)CO), formed after (12)CO (13)CO coadsorption, display bands at 2192 and 2131 cm(-1), in excellent agreement with the theoretically calculated values. Pt(2+)-CO species absorb at 2145 cm(-1) and are not able to accept a second CO molecule. Pt(+)-CO carbonyls are characterized by a band at 2111 cm(-1). Under CO equilibrium pressure, these species are converted into dicarbonyls (nu(s) at 2135 cm(-1) and nu(as) at 2101 cm(-1)). The respective mixed-ligand species, Pt(+)((12)CO)((13)CO), manifest bands at 2123 and 2069 cm(-1), in good agreement again with the theory. Different carbonyls of metallic platinum are observed below 2100 cm(-)(1). In addition, weakly adsorbed CO was registered as Na(+)-CO complexes (2177 and 2165 cm(-1)) and Na(+)-OC-Na(+) species (2138 cm(-1)). It was found that during desorption of CO platinum was reduced, ultimately to metal. However, heating in a NO + O(2) mixture leads to reoxidation of the metal particles and restoration of the initial state of the sample. PMID- 16316121 TI - Interaction between poly[(R)-3-hydroxybutyrate] depolymerase and biodegradable polyesters evaluated by atomic force microscopy. AB - Adsorption of PHB depolymerase from Ralstonia pickettii T1 to biodegradable polyesters such as poly[(R)-3-hydroxybutyrate] (PHB) and poly(l-lactic acid) (PLLA) was investigated by atomic force microscopy (AFM). The substrate-binding domain (SBD) with histidines within the N-terminus was prepared and immobilized on the AFM tip surface via a self-assembled monolayer with a nitrilotriacetic acid group. Using the functionalized AFM tips, the force-distance measurements for polyesters were carried out at room temperature in a buffer solution. In the case of AFM tips with immobilized SBD and their interaction with polyesters, multiple pull-off events were frequently recognized in the retraction curves. The single rupture force was estimated at approximately 100 pN for both PLLA and PHB. The multiple pull-off events were recognized even in the presence of a surfactant, which will prevent nonspecific interactions, but reduced when using polyethylene instead of polyesters as a substrate. The present results provide that the PHB depolymerase adsorbs specifically to the surfaces of polyesters and that the single unbinding event evaluated here is mainly associated with the interaction between one molecule of SBD and the polymer surface. PMID- 16316122 TI - Interactions between nonpolar surfaces coated with the nonionic surfactant n dodecyl-beta-D-maltoside. AB - The forces acting between nonpolar surfaces coated with the nonionic surfactant n dodecyl-beta-D-maltoside (beta-C(12)G(2)) were investigated at concentrations below and above the critical micelle concentration. The long-range and adhesive forces were measured with a bimorph surface force apparatus (MASIF). It was found that the effect of hydrodynamic interactions had to be taken into account for an accurate determination of the short-range static interactions. The results were compared with disjoining pressure versus thickness curves that were obtained earlier with a thin film pressure balance (TFPB). This comparison led to the conclusion that the charges observed at the air-water interface are not due to charged species present in the surfactant sample. In addition, it was observed that the stability of thin liquid films crucially depends on the surfactant's bulk concentration (c) and thus on the packing density in the adsorbed layer. The force barrier preventing removal of the surfactant layer from between two solid liquid interfaces increases with increasing c, while for foam films it is the stability of the Newton black film that increases with c. Finally, the results obtained for beta-C(12)G(2) were compared with those obtained for the homologue n decyl-beta-d-maltoside (beta-C(10)G(2)) as well as with those obtained for nonionic surfactants with polyoxyethylene moieties as polar groups. PMID- 16316123 TI - Motion of a drop on a solid surface due to a wettability gradient. AB - The hydrodynamic force experienced by a spherical-cap drop moving on a solid surface is obtained from two approximate analytical solutions and used to predict the quasi-steady speed of the drop in a wettability gradient. One solution is based on approximation of the shape of the drop as a collection of wedges, and the other is based on lubrication theory. Also, asymptotic results from both approximations for small contact angles, as well as an asymptotic result from lubrication theory that is good when the length scale of the drop is large compared with the slip length, are given. The results for the hydrodynamic force also can be used to predict the quasi-steady speed of a drop sliding down an incline. PMID- 16316124 TI - Morphology transitions in nonionic surfactant adsorbed layers near their cloud points. AB - The structure of adsorbed layers of several polyoxyethylene alkyl ether (C(n)E(m)) nonionic surfactants on silica and graphite surfaces has been imaged using atomic force microscopy as a function of temperature up to their cloud points. For all surfactants with a cloud point within the experimentally accessible range, the adsorbed layer morphology on silica evolved from globules at low temperatures first into rods and then a mesh with increasing temperature. This mesh structure was retained even when the solutions were heated above their cloud points into the two-phase coexistence region. Only C(12)E(3) was observed to form a laterally unstructured bilayer. On graphite, all surfactants formed straight, parallel hemicylinders at all temperatures examined. PMID- 16316125 TI - Remarkable reduction of irreversible fouling and improvement of the permeation properties of poly(ether sulfone) ultrafiltration membranes by blending with pluronic F127. AB - Hydrophilic modification of ultrafiltration membranes was achieved through blending of Pluronic F127 with poly(ether sulfone) (PES). The chemical composition and morphology changes of the membrane surface were confirmed by water contact angle, X-ray photoelectron spectroscopy, scanning electron microscopy, and protein adsorption measurements. The decreased static water contact angle with an increase in the Pluronic F127 content indicated an increase of surface hydrophilicity. XPS analysis revealed enrichment of PEO segments of Pluronic F127 at the membrane surface. The apparent protein adsorption amount decreased significantly from 56.2 to 0 microg/cm(2) when the Pluronic F127 content varied from 0% to 10.5%, which indicated that the blend membrane had an excellent ability to resist protein adsorption. The ultrafiltration experiments revealed that the Pluronic F127 content had little influence on the protein rejection ratio and pure water flux. Most importantly, at a high Pluronic F127 content membrane fouling, especially irreversible fouling, has been remarkably reduced. The flux recoveries of blend membranes reached as high as 90% after periodic cleaning in three cycles. PMID- 16316126 TI - Separating surface and solvent effects and the notion of critical adsorption energy in the adsorption of phenolic compounds by activated carbons. AB - A modified form of the Freundlich equation in which the solute equilibrium concentration is normalized with respect to the solute solubility is analyzed and applied to adsorption isotherms of phenol, 4-nitrophenol, 4-chlorophenol, and 2 chlorophenol at different values of pH on commercial activated carbon before and after oxidation. The analysis confirms the importance of normalizing the solute equilibrium concentration when analyzing the adsorption isotherms, and it is suggested that a parameter, K(F10), obtained by taking 10% solubility as the reference point when applying the Freundlich equation, is probably the best comparative estimate of the relative adsorption capacity of the carbon for different phenolic compounds. In combination with the Freundlich exponent, n(F), estimates of the adsorption capacity at any other reference point can then be obtained. Analysis of the experimental results also indicates a need to distinguish between two regimes of adsorption, characterized by an adsorption energy, E(ads), greater than or less than a critical value, E(ca). When E(ads) > E(ca), the shape of the adsorption isotherm is determined by solute-solid interactions. On the other hand, when E(ads) < E(ca), solute-solution interactions become more important. PMID- 16316127 TI - Ring-shaped morphology in solution-cast polystyrene-poly(methyl methacrylate) block copolymer thin films. AB - We report observation of ring-shaped morphology formed in thin films of a cylinder-forming polystyrene-b-poly(methyl methacrylate) (PS-b-PMMA) diblock copolymer cast from 1,1,2,2-tetrachloroethane (Tetra-CE) solution via relatively fast solvent evaporation rates, in which Tetra-CE is a good solvent for both blocks but preferential affinity for the minority PMMA block. We studied the microstructure of a set of solution-cast block copolymer films dried with different solvent evaporation rates, R. The control with different R leads to keeping microstructures in different solution concentrations (phi) and bringing mechanical strain fields with different strength in the film, for which faster evaporation rates result in microstructures of lower solution concentrations and mechanical strain fields of higher strength. As R decreases from rapid evaporation (approximately 0.1 mL/h), the film microstructure evolved from an intermediate ringlike morphology sequentially to ring-shaped morphologies including loose and tight rings and then inverted phase of PS spheres in a PMMA matrix and finally reached the equilibrium phase, namely, cylinders of PMMA in a PS matrix. In view of the influence of the film constraints, the microstructure of a film with a terraced free surface profile has been examined. The results indicate that the ring-shaped morphology can form as long as the film thickness is larger than a critical value of about one microdomain spacing. In the case where the film thickness is larger than that value, the nature of solvent and the kinetics of solvent evaporation are shown to be mainly responsible for the ring shaped morphology formation. PMID- 16316128 TI - Dielectric analysis of a nanoscale particle in an aqueous solution of low electrolyte concentration. AB - The dielectric spectra of aqueous suspensions of nanoscale silica particles (8 and 24 nm) with low electrolyte concentrations were investigated as a function of the particle concentration. Obvious dispersions observed in the frequency range of 10-10(5) kHz are explained by the multiple effects of the interfacial polarization and the polarization of counterions by using a two-step model and the corresponding dielectric analytical method arising from the combination of Hanai's method and O'Konski's theory. The phase parameters, which reflect the inner properties of constituent phases of the system, are calculated and discussed in detail. The validity of the two-step model was tested in terms of the standard electrokinetic model deduced from pure theories. PMID- 16316129 TI - Self-assembly of model DNA-binding peptide amphiphiles. AB - Peptide amphiphiles combine the specific functionality of proteins with the engineering convenience of synthetic amphiphiles. These molecules covalently link a peptide headgroup, typically from an active fragment of a larger protein, to a hydrophobic alkyl tail. Our research is aimed at forming and characterizing covalently stabilized, self-assembled, peptide-amphiphile aggregates that can be used as a platform for the examination and modular design and construction of systems with engineering biological activity. We have studied the self-assembly properties of a model DNA-binding amphiphile, having a GCN4 peptide as the headgroup and containing a polymerizable methacrylic group in the tail region, using a combination of small-angle X-ray scattering, small-angle neutron scattering, and cryo- transmission electron microscopy. Our results reveal a variety of morphologies in this system. The peptide amphiphiles assembled in aqueous solution to helical ribbons and tubules. These structures transformed into lamella upon DNA binding. In contrast with common surfactants, the specific interaction between the headgroups seems to play an important role in determining the microstructure. The geometry of the self-assembled aggregate can be controlled by means of adding a cosurfactant. For example, the addition of SDS induced the formation of spherical micelles. PMID- 16316130 TI - Interaction of azide ion with hemin and cytochrome c immobilized on Au and Ag nanoparticles. AB - This paper presents a set of investigations on the binding of a metabolic inhibitor, azide with prosthetic heme group of biomolecules, hemin chloride (Hem) and cytochrome c (Cyt c) immobilized on Au and Ag nanoparticles. A variety of spectroscopic tools have been used to understand the chemistry occurring on the nanoparticle surface. While the nature of binding of the model system, hemin has been investigated by UV-visible, fluorescence, FTIR, and Raman spectroscopies, the azide binding has been studied in detail by MALDI-TOF MS. Hemin binding on the nanoparticle surface occurs through the carboxylic acid groups. The hemin N(3) adduct on the nanoparticle surface has been detected by mass spectrometry and its fragments have been studied by post source decay analysis. The chemistry of hemin on the nanoparticle surface has been compared with that of the protein, Cyt c. Azide binding of Cyt c requires thermal activation due to reduced accessibility of the heme center, unlike in the case of hemin. The binding chemistry is similar for free Cyt c and Cyt c bound to the nanoparticles. PMID- 16316131 TI - Proteolytic cleavage reveals interaction patterns between silica nanoparticles and two variants of human carbonic anhydrase. AB - To characterize the sites on the protein surface that are involved in the adsorption to silica nanoparticles and the subsequent rearrangements of the protein/nanoparticle interaction, a novel approach has been used. After incubation of protein with silica nanoparticles for 2 or 16 h, the protein was cleaved with trypsin and the peptide fragments were analyzed with mass spectrometry. The nanoparticle surface area was in 16-fold excess over available protein surface to minimize the probability that the initial binding would be affected by other protein molecules. When the fragment patterns obtained in the presence and absence of silica nanoparticles were compared, we were able to characterize the protein fragments that interact with the surface. This approach has allowed us to identify the initial binding sites on the protein structure and the rearrangement of the binding sites that occur upon prolonged incubation with the surface. PMID- 16316132 TI - Probing the conformational features of a phage display polypeptide sequence directed against single-walled carbon nanohorn surfaces. AB - Single-walled carbon nanohorns (SWNHs) are interesting carbon nanostructures that have applications to science and technology. Using M13 phage display technology, polypeptides directed again SWNHs surfaces have been created for a number of nanotechnology and pharmaceutical purposes, yet the molecular mechanism of polypeptide sequence interaction and binding to SWNHs surfaces is not known. Recently, we identified a linear 12-AA M13 phage pIII sequence, NH-12-5-2 (DYFSSPYYEQLF), that binds with high affinity to SWNHs surfaces. To probe the structure of this pIII tail polypeptide further, we investigated the conformation of a model peptide representing the 12 AA NH-12-5-2 sequence. At neutral pH, the NH-12-5-2 model polypeptide is conformationally labile and exhibits two-state conformational exchange involving the D1-S5 N-terminal segment. Simultaneous with this conformational exchange process is the observation that the P6 residue exhibits imido ring conformational variation. In the presence of the structure stabilizing solvent, TFE, or at pH 2.5, both the exchange process and Pro ring motion phenomena disappear, indicating that the structure of this peptide sequence can be stabilized by extrinsic factors. Interestingly, we observe NMR parameters (ROEs, (3)J coupling constants) for NH-12-5-2 in 90% v/v TFE that are consistent with the presence of a partial helical structure, similar to what was observed at low pH in our earlier CD experiments. We conclude that the NH-12-5-2 model polypeptide sequence possesses an inherent conformational instability that involves the D1-S5 sequence segment and the P6 residue but that this instability can be offset by extrinsic factors (e.g., charge neutralization, imido ring interconversion, and hydrophobic-hydrophobic interactions). These nonbonding interactions may play a role in the recognition and binding of this phage sequence region to SWNHs surfaces. PMID- 16316133 TI - Layer-by-layer assembly of nacre-like nanostructured composites with antimicrobial properties. AB - In a recent report, we have presented the layer-by-layer (LBL) assembly of a biomimetic nanostructured composite from Na(+)-montmorillonite clay nanosheets and poly(diallylmethylammonium chloride) (Tang, Z.; Kotov, N.; Magonov, S.; Ozturk, B. Nat. Mater. 2003, 2, 413). The structure, deformation mechanism, and mechanical properties of the material are very similar to those of natural nacre and lamellar bones. This fact prompts further investigation of these composites as potential bone implants. LBL assembly affords preparation of multifunctional composites, and here we demonstrate that not only mechanical strength, but also antibacterial activity, can be introduced in these implantable materials by alternating clay layers with starch-stabilized silver nanoparticles. The resulting composite showed excellent structural stability with no detectable levels of silver lost over a 1 month period. Evaluation of the antibacterial properties showed almost complete growth inhibition of E. coli over an 18 h period. The amount of silver eluted from the LBL composite over a 1 month period was determined to be only 0.5-3.0 microg/L. This concentration of silver did not prevent the growth of the mammalian tissue cultures. The LBL composite has shown biocompatibility with the human osteoblast cell line. PMID- 16316134 TI - Light-induced surface wettability of a tethered DNA base. AB - We show that the DNA base thymine and other uracil derivatives, when alkylated with a hydrocarbon chain and assembled at a gold interface, dimerize when subjected to UV irradiation. The process is robust and reversible and is accompanied by a substantial decrease in wettability as well as a marked decrease in acidity constant when the dimer is formed. There is a concerted molecular reorientation that accompanies photodimerization, with the dimer displaying a marked affinity for the gold surface. The spatial requirements for this reorganization are satisfied during dimerization by the reduction in intermolecular distance that occurs when the cyclobutane ring is formed between adjacent bases. The structural changes observed here for a tethered DNA base provide a direct route for exploring reactions, in two dimensions, that are of central interest in biology. PMID- 16316135 TI - Correlation between the structure of water in the vicinity of carboxybetaine polymers and their blood-compatibility. AB - The structure and hydrogen bonding of water in the vicinity of carboxybetaine homopolymer (poly[1-carboxy-N,N-dimethyl-N-(2'-methacryloyloxyethyl)methanaminium inner salt] (PolyCMB), and a random copolymer of CMB and n-butyl methacrylate, Poly(CMB-r-BMA), with various molecular weights were analyzed in their aqueous solutions and thin film with contours of O-H stretching of Raman and attenuated total reflection infrared (ATR-IR) spectra, respectively. The relative intensity of the collective band (C value) corresponding to a long-range coupling of O-H stretchings of the Raman spectra for aqueous solution of Poly(CMB-r-BMA) was very close to that for pure water, which is in contrast with the smaller C value in aqueous solution of ordinary polyelectrolytes. The number of hydrogen bonds collapsed by the presence of one monomer residue (N(corr) value) of PolyCMB and Poly(CMB-r-BMA) (CMB, 45 mol %) (M(w), 1.14 x 10(4) and 1.78 x 10(4), respectively) could be calculated from the C value. The N(corr) values were much smaller than those for ordinary polyelectrolytes and close to those for nonionic water-soluble polymers such as poly(ethylene glycol) and poly(N vinylpyrrolidone). Furthermore, a water-insoluble Poly(CMB-r-BMA) with a large BMA content (M(w) = 347 kD, CMB 27 mol %) could be cast as a thin film (thickness, ca. 10 microm) on a ZnSe crystal for the ATR-IR analyses. At an early stage of sorption of water into the Poly(CMB-r-BMA) film, the O-H stretching band of IR spectra for the water incorporated in the film was similar to that for free water, which is in contrast with the drastic change in the O-H stretching band of water incorporated in polymer films such as poly(methyl methacrylate) (PMMA) and poly(n-butyl methacrylate) (PBMA). The theoretical vibrational frequency for water molecules hydrating a betaine molecule calculated by using a density functional method supported the experimental results. The adhesion of human platelets to Poly(CMB-r-BMA) films was much less than that to PMMA and PBMA. With an increase in the content of CMB residue, the number of platelets adhered to the Poly(CMB-r-BMA) film drastically decreased and then gradually increased, probably due to the increase in the roughness of the film surface. These results suggest that the carboxybetaine monomer residues with a zwitterionic structure do not significantly disturb the hydrogen bonding between water molecules in both aqueous solution and thin film systems, resulting in the excellent blood compatibility of the carboxybetaine polymers. PMID- 16316136 TI - Influence of a neoglycolipid and its PEO-lipid moiety on the organization of phospholipid monolayers. AB - The surface properties of the neoglycolipid (GlcNAcE(3)G(28)) and of its PEO lipid (E(3)G(28)) moiety mixed with phospholipids (dipalmitoylphosphatidylcholine, DPPC; distearoylphosphatidylcholine, DSPC; diarachidoylphosphatidylcholine, DAPC; and dibehenoylphosphatidylcholine, DBPC) were studied in Langmuir monolayers at various mixture compositions and surface pressures. The pi-A isotherms of the pure compounds revealed that because of the presence of the sugar group in its molecule, GlcNAcE(3)G(28) collapsed at a higher surface pressure and occupied a larger molecular area than the PEO-lipid moiety. It was also observed that the presence of the PEO-lipid (E(3)G(28)) in the mixtures triggered a strong alteration of both phospholipid pi-A isotherm profiles and surface diffraction spectra, an indication that the disordering of the initially structured phospholipid monolayers took place. Unlike E(3)G(28), GlcNAcE(3)G(28) did not disorganize phospholipid monolayers but generated a partial segregation of the film-forming components. The calculated excess free energies of mixing (DeltaG(exc)) for GlcNAcE(3)G(28)-phospholipid mixtures enabled us to predict the stability of such systems. PMID- 16316137 TI - Lipase surface diffusion studied by fluorescence recovery after photobleaching. AB - We have analyzed surface diffusion properties of a variant of Thermomyces lanuginosa lipase (TLL) on hydrophilic silica and silica methylated with dichlorodimethylsilane (DDS) or octadecyltrichlorosilane (OTS). For this study a novel method for analysis of diffusion on solid surfaces was developed. The method is based on fluorescence recovery after photobleaching using confocal microscopy. When a rectangular area of the sample was photobleached, fluorescence recovery could be analyzed as one-dimensional diffusion, resulting in simplified mathematical expressions for fitting the data. The method was initially tested by measuring bovine serum albumin diffusion on glass, which led to a diffusion coefficient in good correspondence to earlier reports. For the analysis of TLL diffusion, ellipsometry data of TLL adsorption were used to calibrate fluorescence intensity to surface density of lipase, enabling measurements of the diffusion coefficient at different surface densities. The average diffusion coefficient was calculated in two time intervals after adsorption. Mobile fraction and diffusion coefficient were lowest on the OTS surface, when extrapolated to infinite surface dilution. Moreover, the diffusion rate decreased with time on the hydrophobic surfaces. Our observations can be explained by the surface dependence on the distribution of orientations and conformations of adsorbed TLL, where the transition from the closed to the catalytically active open and more hydrophobic structure is important. PMID- 16316138 TI - An aqueous-based surface modification of poly(dimethylsiloxane) with poly(ethylene glycol) to prevent biofouling. AB - We report a simple modification of poly(dimethylsiloxane) (PDMS) surfaces with poly(ethylene glycol) (PEG) through the adsorption of a graft copolymer, poly(l lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) from aqueous solution. In this approach, the PDMS surface was treated with oxygen plasma, followed by immersion into aqueous solution containing PLL-g-PEG copolymers. Due to the hydroxyl/carboxylic groups generated on the PDMS surface after oxygen plasma, the polycationic PLL backbone is attracted to the negatively charged surface and PEG side chains exhibit an extended structure. The PEG/aqueous interface generated in this way revealed a near-perfect resistance to nonspecific protein adsorption as monitored by means of optical waveguide lightmode spectroscopy (OWLS) and fluorescence microscopy. PMID- 16316139 TI - E-cadherin tethered to micropatterned supported lipid bilayers as a model for cell adhesion. AB - Cell-cell adhesion is a dynamic process requiring recruitment, binding, and reorganization of signaling proteins in the plane of the plasma membrane. Here, we describe a new system for investigating how this lateral mobility influences cadherin-based cell signaling. This model is based on tethering of a GPI-modified E-cadherin protein (hEFG) to a supported lipid bilayer. In this report, membrane microfluidics and micropatterning techniques are used to adopt this tethered protein system for studies with the anchorage-dependent cells. As directly formed from proteoliposomes, hEFG exhibits a diffusion coefficient of 0.6 +/- 0.3 microm(2)/s and mobile fraction of 30-60%. Lateral structuring of the supported lipid bilayer is used to isolate mobile proteins from this mixed mobile/immobile population, and should be widely applicable to other proteins. MCF-7 cells seeded onto hEFG-containing bilayers recognize and cluster this protein, but do not exhibit cell spreading required for survival. By micropatterning small anchors into the supported lipid bilayer, we have achieved cell spreading across the bilayer surface and concurrent interaction with mobile hEFG protein. Together, these techniques will allow more detailed analysis of the cellular dynamics involved in cadherin-dependent adhesion events. PMID- 16316140 TI - Homogeneously distributed CdS nanoparticles in nafion membranes: preparation, characterization, and photocatalytic properties. AB - Stable crystalline CdS nanoparticles were synthesized in Nafion ionomer membranes by using thioacetamide (TAA) as a nonionic precursor. Unlike the ionic precursors such as Na(2)S, TAA could diffuse into the cationic-exchangeable ionomer membranes much more uniformly. This led to the formation of homogeneously distributed CdS nanoparticles in the Nafion membranes, which was confirmed by elemental mapping with energy-dispersive X-ray (EDAX) analysis. Results from the characterizations on the physical properties, the chemical stability, and the photocatalytic properties of these CdS nanoparticles embedded in Nafion membranes are presented and discussed. The parallel data from the CdS nanoparticles in Nafion membranes prepared from the ionic Na(2)S precursor are also shown for comparison. PMID- 16316141 TI - Influence of initial order on the microscopic mechanism of electric field induced alignment of block copolymer microdomains. AB - We investigate the mechanism of microdomain orientation in concentrated block copolymer solutions exposed to a dc electric field by in situ synchrotron small angle X-ray scattering (SAXS). As a model system, we use concentrated solutions of a lamellar polystyrene-b-polyisoprene block copolymer in toluene. We find that both the microscopic mechanism of reorientation and the kinetics of the process strongly depend on the initial degree of order in the system. In a highly ordered lamellar system with the lamellae being aligned perpendicular to the electric field vector, only nucleation and growth of domains is possible as a pathway to reorientation and the process proceeds rather slowly. In less ordered samples, grain rotation becomes possible as an alternative pathway, and the process proceeds considerably faster. The interpretation of our finding is strongly corroborated by dynamic self-consistent field simulations. PMID- 16316142 TI - Hydrophobic dendrimers as templates for au nanoparticles. AB - We report the synthesis, characterization, and extraction of Au dendrimer encapsulated nanoparticles (DENs) prepared in organic solvents. DENs composed of 31 and 55 Au atoms were prepared using organic solvents and poly(amidoamine) (PAMAM) dendrimer templates modified on their periphery with dodecyl groups. The spectral and microscopic properties of the resulting materials were identical to those prepared using water-soluble PAMAM dendrimers. It was possible to extract the organic-soluble DENs into water using the water-soluble thiols tiopronin and glutathione. The properties of the resulting monolayer-protected clusters were nearly identical to those of the precursor DENs. A mechanistic model for the extraction process is discussed. The synthetic methodology reported here provides a convenient method for preparing DENs of non noble metals such as Ni and Fe. PMID- 16316143 TI - Self-supported particle-track-etched polycarbonate membranes as templates for cylindrical polypyrrole nanotubes and nanowires: an X-ray scattering and scanning force microscopy investigation. AB - Self-supported particle-track-etched polycarbonate membranes with nearly perfect cylindrical pores are used for the preparation of similarly perfect cylindrical polypyrrole nanowires and nanotubes. A complete investigation of the structural properties that result at different stages of the preparation route of polypyrrole nanowires and nanotubes is based on a combination of real and reciprocal space techniques. Nanoporous membranes with nominal pore size ranging from 5 to 150 nm and pore density up to 10(9) pores/cm(2) made from 21-microm thick polycarbonate films are used. Polypyrrole nanotubes or nanowires are synthesized inside the pores. A real-space picture of the nanomaterial results from scanning force microscopy (SFM) on ultrathin sections made in two directions to obtain structures in the sample surface as well as perpendicular to the surface. From a model-based fit to the small-angle and ultra-small-angle X-ray scattering (SAXS/USAXS) data, the geometric pore structure is obtained and compared to values determined with scanning electron microscopy (SEM). Nanopores, nanowires, and nanotubes are described by uniform solid cylinders or hollow tubes, which are oriented highly parallel to each other and exhibit a small size distribution. Below a critical pore diameter, solid nanowires are produced whereas above this limit hollow nanotubes result. PMID- 16316144 TI - Swollen poly(dimethylsiloxane) (PDMS) as a template for inorganic morphologies. AB - We report a series of silica, titania, and zirconia microstructures synthesized within swollen poly(dimethylsiloxane) (PDMS). Voids created by solvent-swelling the polymer are used to template the product. The inorganic morphologies range from spheres to networks, depending upon the nature of the polymer, its degree of swelling, and the synthetic conditions. Organic solvents as well as pure metal alkoxide liquids have been used to swell the polymer. Once the alkoxide precursor is inside the swollen polymer, water is introduced to bring about hydrolysis and condensation polymerization. The product is a textured metal oxide within a PDMS matrix. Scanning electron microscopy (SEM), optical microscopy, nuclear magnetic resonance (NMR), and powder X-ray diffraction (PXRD) were used to characterize the products. Microstructures formed in this manner have potential use as an inexpensive route to catalysts, fillers, capsules, or membranes for separations. PMID- 16316145 TI - Surface functionalization of ordered mesoporous carbons--a comparative study. AB - Hexagonally structured mesoporous carbons C15 and CMK-5 and cubically structured carbon C48 were synthesized using ordered silica SBA-15 and MCM-48 as templates and carbon precursors of different structures. The surfaces of these ordered carbons were chemically functionalized by employing an approach, in which the selected diazonium compounds were in situ generated and reacted with the carbon frameworks of the mesoporous carbons. The aromatic organic molecules containing chlorine, ester, and alkyl groups were covalently attached to the surface of these ordered mesoporous carbons. The presence of functional groups on the modified carbons was confirmed with Fourier transform infrared spectroscopy, thermogravimetric analysis, and nitrogen adsorption. The BET-specific surface area and the pore width of ordered carbons were significantly reduced, whereas the primary structure of these ordered carbons and their unit cells were intact. Basically, the density of grafted functional groups is related to the specific surface area of the sample, particularly the surface area of mesopores. The surface functionalization reaction takes place only on the external surface of carbon C15, while it occurs on both of the internal and external surface of CMK-5 carbon with the nanopipe structure. The presence of the micropores in CMK-5 carbon should be responsible for its lower grafting density because the small micropores are inaccessible in the reaction. It was also proposed that the preferred adsorption/reaction in C48 may be related to the observed unsymmetrical degradation of the XRD patterns for the functionalized C48 samples. The chemical modification process considerably reduced the primary mesopores in these ordered carbons by approximately 1-1.5 nm, affording carbons with micropores in the cases of C15 and C48, and mixed micropores and small mesopores in the case of CMK-5. A grafting density of approximately 0.9-1.5 micromol/m(2) was achieved under current research. PMID- 16316146 TI - Electrochemical control of the interactions of polymers and redox-active surfactants. AB - We report the characterization of aqueous solutions (0.1 M Li(2)SO(4)) of dilute ethyl(hydroxyethyl) cellulose (EHEC) mixed with the redox-active surfactant (11 ferrocenylundecyl)trimethylammonium bromide (FTMA) by measurements of clouding temperatures and dynamic light scattering. The investigation sought to test the hypothesis that FTMA, which forms micelles in aqueous solution in its reduced state but not in its oxidized state, would permit reversible control over the formation of polymer-surfactant complexes in solution. Our measurements revealed that low concentrations of reduced FTMA enhance aggregation, whereas high concentrations of reduced FTMA disperse polymer-surfactant aggregates. This behavior is qualitatively similar to both dodecyltrimethylammonium bromide and sodium dodecyl sulfate and reflects cooperative interactions between FTMA and EHEC. In contrast, oxidized FTMA was found not to promote EHEC aggregation at low concentrations of oxidized FTMA nor disperse EHEC aggregates at high concentrations. Measurements of dynamic light scattering revealed that the reduction of oxidized FTMA in solutions of EHEC containing 0.1-0.3 mmolal FTMA causes an increase in the sizes of polymer-surfactant aggregates of more than 1 order of magnitude. Our cloud point measurements also revealed that clouding can be induced isothermally via the electrochemical reduction of oxidized FTMA at room temperature at FTMA concentrations between 0.4 and 10 mmolal. In contrast, at concentrations of FTMA greater than 15 mmolal, the reduction of oxidized FTMA induces the clearing of EHEC solutions. We conclude that aggregation of EHEC in dilute solutions can be controlled by the manipulation of the oxidation state of FTMA. PMID- 16316147 TI - Gold nanoparticle localization at the core surface by using thermosensitive core shell particles as a template. AB - We report novel thermosensitive hybrid core-shell particles via in situ gold nanoparticle formation using thermosensitive core-shell particles as a template. This method for the in situ synthesis of gold nanoparticles with microgel interiors offers the advantage of eliminating or significantly reducing particle aggregation. In addition, by using thermosensitive microgel structures in which the shell has thermosensitive and gel properties in water--whereas the core itself is a water-insoluble polymer--we were able to synthesize the gold nanoparticles only at the surface of the core, which had reactive sites to bind metal ions. After the gold nanoparticles were synthesized, electroless gold plating was carried out to control the thickness of the gold nanoshells. The dispersions of the obtained hybrid particles were characterized by dynamic light scattering and UV-vis absorption spectroscopy, and the dried particles were also observed by electron microscopy. Adaptation of the technique shown here will create a number of applications as optical, electronic, and biomedical functional materials. PMID- 16316148 TI - Dynamics and density profile of water in nanotubes as one-dimensional fluid. AB - The transport and structural properties of water confined in nanotubes with different diameters were studied by molecular dynamics (MD) simulation. The effects of pore size, molecule-wall interaction, and the helicity of CNT on the diffusivity, thermal conductivity, and shear viscosity as well as density profile were analyzed. For diffusivity, in model NT > in armchair CNT > in zigzag CNT at similar conditions. However in contrast to the diffusivity, the thermal conductivity and the shear viscosity increase as the pore size decreases, in zigzag CNT > in armchair CNT > (or approximately ) in model NT. The ordered layer distribution of water molecules in nanotubes is clear. It suggests the structure of fluid in the zigzag CNTs is more ordered, and more solidlike. In the nanotubes, where the molecule and the pore dimensions are of similar order of magnitude, the nature of water-water and water-wall interactions, the confinement effect of space, and the helicity of CNT become more significant. PMID- 16316149 TI - Anionic microemulsion-mediated low temperature synthesis of anisotropic silicalite-1 nanocrystals. AB - The low-temperature (368 K) synthesis of silicalite-1 nanocrystals in anionic microemulsions is reported. In the presence of AOT/isooctane mixtures silicalite 1 nanocrystals can be formed that are coffin-shaped and approximately 100 x 40 x 200 nm in size. This is in contrast to samples made without the microemulsion under the same conditions where irregular spherical particles approximately 100 nm in diameter are formed. The current work shows that, in contrast to previous work in this area, the anionic microemulsions cannot stabilize colloidal silica due to the strong repulsive electrostatic forces between the anionic silicate species and the surfactant headgroup. The crystal morphology of the silicalite-1 obtained is also shown to be sensitive to the surfactant identity as syntheses using SDS/heptane/butanol mixtures lead to different morphologies. It is also possible to uncouple zeolite nucleation from growth in these systems. This was demonstrated by adding a solution containing 25 nm silicalite-1 nanocrystals to the AOT/isooctane mixture, which leads to large micron-sized spheres of silicalite-1 containing large mesopores. This report demonstrates that anionic microemulsions lead to fundamentally different crystal habits than the nonionic or cationic microemulsions investigated previously. The future outlook for the use of microemulsion-mediated zeolite growth is also discussed. PMID- 16316150 TI - Dielectrophoretic force on a sphere near a planar boundary. AB - The small gap distance separating a spherical colloidal particle in electrophoretic motion from a planar nonconducting surface is a required parameter for calculating its electrophoretic mobility. In the presence of an externally applied electric field, this gap distance is determined by balancing the van der Waals, electrical double layer interaction, and gravitational forces with a dielectrophoretic (DEP) force. Here, the DEP force was determined analytically by integration of the Maxwell stress over the surface of the particle. The account of this force showed that its previous omission from the analysis always resulted in underpredicted gap distances. Furthermore, the DEP force dominated under conditions of low particle density or high electric field strength and led to much higher gap distances on the order of a few microns. In one particular case, a combination of low particle density and small particle size produced two possible equilibrium gap distances for the particle. However, the particle was unstable in the second equilibrium position when subjected to small perturbations. In general, larger particles had smaller gap sizes. The effects of four other parameters on gap distance were studied, and gap distances were found to increase with lower particle density, higher electric field strength, higher particle and wall zeta potentials, and lower Hamaker constants. Retardation effects on van der Waals attraction were considered. PMID- 16316151 TI - Steric repulsion between internal aqueous droplets and the external aqueous phase in double emulsions. AB - A theoretical model for analyzing the steric repulsion energy between internal aqueous droplets and the external aqueous phase in double emulsions, which results from the steric interaction between the surfactant molecules adsorbed at the two interfaces, has been established. The steric interaction is dependent on the separation distance between the internal aqueous droplets and the external aqueous phase, the thicknesses of the two adsorbed surfactant layers, and the size of the internal aqueous droplets and the oil globules, all of which determine the extent of the compression of the adsorbed surfactant molecules. The thickness of each of the two surfactant layers have the same effect on the steric repulsion, and stronger steric interaction can be achieved with thicker adsorbed layers, which can effectively prevent coalescence between the internal aqueous droplets and the external aqueous phase. Increasing the internal aqueous droplet size can produce stronger steric repulsion; however, larger oil globules will weaken the steric repulsion, indicating that a more stable double-emulsion system can be achieved by preparing the system with smaller oil globules and larger internal aqueous droplets. PMID- 16316154 TI - Surface plasmon resonance imaging measurements of the inhibition of Shiga-like toxin by synthetic multivalent inhibitors. AB - A variety of new methodologies to pattern biomolecules on surfaces and to detect binding events are currently being developed for high-throughput assay applications. Carbohydrates serve as attachment sites for toxins, bacteria, and viruses. Immobilized carbohydrate units can thus be used to directly detect these agents or as a platform for inhibitor assessment. In this work, modified glycosides were patterned on gold surfaces to monitor the binding of the homopentameric B5 cell-recognition subunit of the Shiga-like toxin (SLT). Binding was detected with the label-free method of surface plasmon resonance (SPR) imaging. Two synthetic multivalent inhibitors were used in order to effect inhibitory binding, and SPR imaging is presented as a simple alternative to ELISA for the study of toxin inhibition. In contrast to existing methods for the study of carbohydrate-protein interactions, in particular ELISA, the use of micropatterned sensor surfaces is shown to be advantageous due to a decrease in complications and manual labor from numerous blocking, washing, and labeling steps. Carbohydrate receptor density on the sensor surface was optimized in order to effect the maximum binding of the SLT. The IC50 values determined were in the low-nanomolar range for each of the two inhibitors studied. PMID- 16316155 TI - Pyrosequencing in a microfluidic flow-through device. AB - To explore genome variation meaningfully, there is a critical need for a high throughput and inexpensive platform for DNA analysis. Pyrosequencing is a nonelectrophoretic bioluminometric DNA sequencing method that uses a four-enzyme mixture reaction to monitor nucleotide incorporation in real time. Currently, the commercialized pyrosequencing technique is limited to a 96-microtiter plate format. However, high throughput and inexpensive pyrosequencing is required to meet the need of screening large numbers of samples. We present here DNA pyrosequencing on a nanoliter-volume microfluidic platform. The microfluidic approach involves the trapping of the DNA on microbeads in an on-chip filter chamber and flow-through of the pyrosequencing reagents to monitor the reaction in real time. Two single-nucleotide polymorphisms were successfully scored to evaluate the microfluidic platform. In addition to significantly reducing reagent costs, microfluidic systems promise to improve the read length by eliminating intermediate product accumulation by constant removal of unincorporated nucleotides and elimination of dilution effects at each reaction cycle in the current plate format. Although only one filter chamber was used in this study, the platform should be readily adaptable to parallel analyses of nanoliter samples using filter chamber arrays to obtain high-throughput DNA analysis. PMID- 16316156 TI - Inhibitor screening using immobilized enzyme reactor chromatography/mass spectrometry. AB - We describe the coupling of capillary-scale monolithic enzyme reactor columns directly to a tandem mass spectrometer for screening of enzyme inhibitors. A two channel nanoLC system is used to continuously infuse substrate or substrate/inhibitor mixtures through the column, allowing continuous variation of inhibitor concentration by simply altering the ratio of flow from the two pumps. In the absence of inhibitor, infusion of substrate leads to formation of product, and both substrate and product ions can be simultaneously monitored in a quantitative manner by MS/MS. The presence of inhibitor leads to a decrease in product and an increase in substrate concentration in the column eluent. Knowing the product/substrate ratio and the total analyte concentration (P + S), the concentration of product eluting, and hence the relative enzyme activity, can be determined. Both IC50 and KI values can then be obtained by direct MS detection of the effect of inhibitors on relative activity. Inhibitor screening is demonstrated using reusable, sol-gel derived, monolithic capillary columns containing adenosine deaminase, directly interfaced to ESI-MS/MS. On-column enzyme activity was assessed by monitoring inosine and adenosine elution. It is shown that the method can be used for automated screening of the effects of compound mixtures on ADA activity and to determine the KI value of the known inhibitor, erythro-9-(2-hydroxy-3-nonyl)adenine, even when the compound is present within a mixture. PMID- 16316157 TI - Microfluidic biosensor for the serotype-specific detection of dengue virus RNA. AB - The development of a microfluidic biosensor with fluorescence detection for the rapid, sensitive, and serotype-specific detection of Dengue virus is presented. The biosensor chip consists of poly(dimethylsiloxane) (PDMS) substrate with fabricated microchannels and a glass substrate used to seal the microchannels. These two substrates are packaged within a pressure-closed Plexiglas housing to provide a watertight reversible sealing at the PDMS-glass interface. The ability to reversibly seal the device permits easy disassembly and quick interchange of the device parts, which is ideal for developmental purposes. The biosensor employs a magnetic bead-based sandwich hybridization system in conjugation with liposome amplification for the specific detection of nucleic acids. The concentrations of the various biosensor components were optimized using a synthesized fragment of Dengue virus RNA. To evaluate the sensitivity of the assay, two detection systems, based on fluorescence measurements of intact and lysed liposomes, were analyzed. The entire analysis was complete within 20 min (including incubation time) with RNA detection limits of 0.125 nM and 50 pM for intact and lysed liposome detection systems, respectively. Subsequently, the biosensor was applied to the analysis of actual RNA obtained from Dengue virus serotypes 1-4. The resulting signals were compared to those obtained using standard electrochemiluminescence detection and shown to correspond perfectly with respect to serotype identification. PMID- 16316158 TI - Production of ultrafine particles by nanosecond laser sampling using orthogonal prepulse laser breakdown. AB - The particle size distribution and composition of nanosecond laser-generated aerosols from brass samples in atmospheric argon has been measured by low pressure impaction and subsequent quantitative analysis of the aerosols by total reflection X-ray fluorescence. Ablation was performed applying a Nd:YAG laser at 1.06 microm both without and with a prepulse plasma breakdown generated by a second Nd:YAG laser at 2-60 micros prior to the ablation pulse. The beam of the prepulse laser had orthogonal direction to the ablation laser beam, and the breakdown was produced 2.5 mm above the ablation spot. Ultrafine aerosol particles (<50 nm) were generated in the double-pulse experiment representing practically the total mass impacted, while in single-pulse ablation the proportion of large particles (>0.1 microm) was dominating. The predominance of ultrafine aerosols in the prepulse experiment indicates that particle formation from vapor-phase condensation is the major mechanism, while the appearance of large particles in single-pulse ablation points at fragmentary evaporation in the laser-produced plasma. It was also shown that the total mass impacted in double pulse ablation increases almost linearly with the power of the prepulse laser. The better atomization and the larger sample mass ablated can be assumed to be the main reasons for the increase of the line intensities in double-pulse laser induced breakdown spectrometry with orthogonal prepulses reported by several research groups. PMID- 16316159 TI - An NMR approach applicable to biomolecular structure characterization. AB - A generalized 2D correlation NMR (GEN2D-NMR) scheme capable of substantially reducing the experimental time for two-dimensional correlation NMR experiments is described. The experimental time used in GEN2D-NMR is shortened to less than one tenth of that required in traditional double Fourier transform 2D-NMR (FT2D-NMR) for a 13C-13C spin diffusion experiment for Nephila edulis spider threads. Experimentally, one needs to acquire many fewer points in GEN2D-NMR than that in FT2D-NMR in the evolution time domain. By keeping other acquisition pulse sequence and parameters the same in both methods, the GEN2D-NMR technique can produce a 2D-NMR spectrum using fewer points along the evolution time domain equivalent to that produced by the FT2D-NMR technique using more points. GEN2D NMR would provide a significant advantage for those molecules that are difficult to highly isotropically label, such as proteins, polypeptides, and polymers, or those which become unstable in a prolonged measurement time. PMID- 16316160 TI - Covalent micropatterning of poly(dimethylsiloxane) by photografting through a mask. AB - A new photografting method to micropattern a covalent surface modification on poly(dimethylsiloxane) (PDMS) provides advantages in simplicity and efficiency. To accomplish the entire process on the benchtop, the PDMS was initially treated with benzophenone dissolved in a water/acetone mixture. This process permitted limited diffusion of the photoinitiator into the PDMS surface. Polymerization of acrylic acid was initiated by exposure of the benzophenone-implanted PDMS to UV radiation through a photomask with a thin aqueous layer of acrylic acid sandwiched between the PDMS and photomask. This procedure resulted in patterned poly(acrylic acid) (PAA) on the PDMS surface. In the modified regions, PAA and PDMS formed an interpenetrating polymer network extending 50 microm into the PDMS with an X-Y spatial resolution of 5 microm. The carboxyl groups of the PAA graft could be derivatized to covalently bond other molecules to the patterned PAA. Two bioanalytical applications of this micropatterned surface were demonstrated: (1) a guide for cell attachment and growth and (2) a substrate for immunoassays. 3T3 cells were shown to selectively localize to modified surface regions where they could be cultured for up to 7 days. Additionally, the micropatterned surface was used to immobilize either protein A or antibody for heterogeneous immunoassays. PMID- 16316161 TI - Engineering camel single-domain antibodies and immobilization chemistry for human prostate-specific antigen sensing. AB - The specificity and affinity characteristics of antibodies make them excellent probes in biosensor applications. Unfortunately, their large size, unstable behavior, and random immobilization properties create numerous problems. The single-domain antigen-binding fragment derived from heavy-chain antibodies of camelids (termed VHH) offers special advantages in terms of size, stability, and ease of generating different antibody constructs. In this study, we show the potential of those VHHs in sensing human prostate-specific antigen (hPSA) by SPR technology. Different VHH constructs were immobilized onto commercial and custom built sensor surfaces by metal chelation, biotin-streptavidin interaction, or covalent coupling. The detection of subnanogram per milliliter hPSA concentrations could be attained on a covalently coupled three-dimensional dextran surface. Moreover, the ratio of different hPSA isoform concentrations could be assessed via a sandwich assay and resulted in the detection of clinically significant antigen concentrations within 15 min. In addition, for the first time, the intrinsic protein stability is presented as an important probe design factor, since our results reveal that higher intrinsic stability offers higher resistance to harsh regeneration conditions. In conclusion, we present VHHs as a novel class of biosensor probes rivaling conventional antibodies and their derived antibody fragments. PMID- 16316162 TI - In vivo fluorescence detection of glucose using a single-walled carbon nanotube optical sensor: design, fluorophore properties, advantages, and disadvantages. AB - In this work, several aspects of in vivo glucose detection using a nanotube-based optical sensor are considered. The optical properties of commonly used organic and nanoparticle fluorescent probes are compared with respect to quantum yield, human tissue penetration, and photobleaching stability. The latter two factors are shown to dominate sensor viability and require a near-infrared nanoparticle fluorophore for practical device operation. The dynamics of a model optical sensor are compared to a flux-measuring electrochemical sensor of equal area using a mathematical simulation of a healthy patient ingesting three predefined meals per day. Both sensors demonstrate an approximately linear response to blood glucose levels. It is shown that the optical sensor, which transduces glucose concentration, not flux, directly is significantly more stable to membrane biofouling. PMID- 16316163 TI - Silicon microfabricated column with microfabricated differential mobility spectrometer for GC analysis of volatile organic compounds. AB - A 3.0-m-long, 150-microm-wide, 240-microm-deep channel etched in a 3.2-cm-square silicon chip, covered with a Pyrex wafer, and coated with a dimethyl polysiloxane stationary phase is used for the GC separation of volatile organic compounds. The column, which generates approximately 5500 theoretical plates, is temperature programmed in a conventional convection oven. The column is connected through a heated transfer line to a microfabricated differential mobility spectrometer. The spectrometer incorporates a 63Ni source for atmospheric-pressure chemical ionization of the analytes. Nitrogen or air transport gas (flow 300 cm(3)/min) drives the analyte ions through the cell. The spectrometer operates with an asymmetric radio frequency (RF) electric field between a pair of electrodes in the detector cell. During each radio frequency cycle, the ion mobility alternates between a high-field and a low-field value (differential mobility). Ions oscillate between the electrodes, and only ions with an appropriate differential mobility reach a pair of biased collectors at the downstream end of the cell. A compensation voltage applied to one of the RF electrodes is scanned to allow ions with different differential mobilities to pass through the cell without being annihilated at the RF electrodes. A unique feature of the device is that both positive and negative ions are detected from a single experiment. The combined microfabricated column and detector is evaluated for the analysis of volatile organic compounds with a variety of functionalities. PMID- 16316164 TI - Direct analysis of pharmaceutical drug formulations using ion mobility spectrometry/quadrupole-time-of-flight mass spectrometry combined with desorption electrospray ionization. AB - A novel approach to the rapid analysis of pharmaceutical drug formulations using hyphenated ion mobility spectrometry (IMS) and time-of-flight mass spectrometry (ToF-MS) that requires no sample pretreatment or chromatographic separation is described. A modified quadrupole time-of-flight mass spectrometer containing an ion mobility drift cell was used for gas-phase electrophoretic separation of ions prior to ToF-MS detection. The generation of sample ions directly from tablets and cream formulations was effected by desorption electrospray ionization (DESI) using a modified electrospray ion source. The analysis of a range of over-the counter and prescription tablet formulations is described, including histamine H2 receptor antagonist (ranitidine), analgesic (paracetamol), opiate (codeine), and aromatase inhibitor anticancer (anastrozole) drugs. The successful determination of active drugs from soft formulations, such as an antiseptic cream (chlorhexidine) and a nicotine-containing skin patch, is also presented. Limits of detection for the active drugs using the DESI/IMS/ToF-MS method fell within the high-picomole to nanomole range. In all cases, the use of ion mobility drift tube separation showed increased selectivity for active drug responses (present as low as 0.14% w/w) over excipient responses such as poly(ethylene glycol). Tandem mass spectrometric analysis of precursor ions separated by IMS allowed positive confirmation of active drugs with little loss of ion mobility efficiency. The ability to analyze hard or soft pharmaceutical formulations directly by DESI combined with ion mobility spectrometry/mass spectrometry in approximately 2 min demonstrates the potential applicability of this novel method to pharmaceutical screening of low-molecular-weight drug formulations with high selectivity over the formulation vehicle. PMID- 16316165 TI - MASPIC: intensity-based tandem mass spectrometry scoring scheme that improves peptide identification at high confidence. AB - Algorithmic search engines bridge the gap between large tandem mass spectrometry data sets and the identification of proteins associated with biological samples. Improvements in these tools can greatly enhance biological discovery. We present a new scoring scheme for comparing tandem mass spectra with a protein sequence database. The MASPIC (Multinomial Algorithm for Spectral Profile-based Intensity Comparison) scorer converts an experimental tandem mass spectrum into a m/z profile of probability and then scores peak lists from potential candidate peptides using a multinomial distribution model. The MASPIC scoring scheme incorporates intensity, spectral peak density variations, and m/z error distribution associated with peak matches into a multinomial distribution. The scoring scheme was validated on two standard protein mixtures and an additional set of spectra collected on a complex ribosomal protein mixture from Rhodopseudomonas palustris. The results indicate a 5-15% improvement over Sequest for high-confidence identifications. The performance gap grows as sequence database size increases. Additional tests on spectra from proteinase-K digest data showed similar performance improvements demonstrating the advantages in using MASPIC for studying proteins digested with less specific proteases. All these investigations show MASPIC to be a versatile and reliable system for peptide tandem mass spectral identification. PMID- 16316166 TI - Enrichment and identification of cysteine-containing peptides from tryptic digests of performic oxidized proteins by strong cation exchange LC and MALDI TOF/TOF MS. AB - The extreme complexity of sample and uninformative fragmentation of peptides in MS/MS experiments are two of several real challenges faced by proteomics. In this work, a strategy aimed at tackling these two problems is presented. Briefly, proteins were first oxidized by performic acid to cleave the disulfide bonds and simultaneously convert cysteine residue into its sulfonic form. Then the resultant sulfonic peptides were enriched by SCX chromatography, exploiting the negative solution charge of sulfonic group. The sulfonic peptide could be easily detected by MALDI-MS in negative mode and showed both enhanced fragmentation efficiency and a simplified spectrum in MALDI-MS/MS experiment in positive mode. The strength of the strategy was demonstrated by applying it to bovine serum albumin. Potential use of the strategy in proteomics was also discussed. PMID- 16316167 TI - Equilibrium sampling through membranes of freely dissolved copper concentrations with selective hollow fiber membranes and the spectrophotometric detection of a metal stripping agent. AB - A sensitive spectrophotometric method for the determination of freely dissolved copper concentrations in aqueous samples after preconcentration with hollow fiber membrane extraction has been developed. The method is based on the equilibrium sampling through a selective membrane into an acceptor solution containing 4 (pyridyl-2-azo)resorcinol (PAR), which serves as stripping agent and metal indicator. Negligible extraction of interferences and equilibrium enrichment of copper allowed for selective spectrophotometric determination of the Cu-PAR complex. Some important extraction parameters such as acceptor composition, shaking, equilibrium time, and sample volume were studied. The optimized methodology showed good linearity in the range of 5-100 microg/L, an enrichment factor of 93, good repeatability and reproducibility (RSDs < 6%, n = 6), and a detection limit of 4 microg/L. The cationic metals Ni2+, C(2+, Cd2+, Fe3+, Pb2+, Zn2+, and Mn2+ were shown not to interfere with the measurement of Cu2+. Measurements on samples containing mixtures of various ligands and cations were in good agreement with theoretically calculated concentrations, and the method was also applied to environmental samples. The developed technique requires less labor and less sophisticated equipment than conventional methods typically based on atomic absorption spectrometry or ICP. PMID- 16316168 TI - Using volatile solvents for ion formation in liquid molecular beam expansion mass spectrometry. AB - Cocrystallization between analyte and matrix is required by matrix-assisted laser desorption/ionization and can represent a significant limitation of the technique. A molecular beam expansion, mass spectrometric method has been developed to explore the possibility of using pure solvents as matrix to avoid cocrystallization. Two kinds of solvent, liquid CS2 and liquid or supercritical CO2, have been studied with 266-nm UV laser irradiation. We successfully ionized a number of compounds, including caffeine, guanine, cholesterol, and mixed fullerenes. Under some conditions, the mass spectra reflect parent radical cations formed by photoionization. Under other conditions, protonated, sodiated (and with CS2 even sulfated) ions are seen reflecting a nonunimolecular ionization process. When UV-transparent CO2 is used as a solvent, only analyte molecules with a UV chromophore are detected. However, with UV-absorbing CS2, we demonstrate ionization of molecules lacking a UV chromophore. This work provides strong evidence that one can form solvent clusters containing analyte, that laser photoionization of the solvent precedes ionization of the analyte, and that solvent evaporation along with the indirect ionization leads to reduced parent ion fragmentation. The exploration of this now demonstrated concept with other solvents would appear fruitful for future work. PMID- 16316169 TI - Mixing crowded biological solutions in milliseconds. AB - In vitro studies of biological reactions are rarely performed in conditions that reflect their native intracellular environments where macromolecular crowding can drastically change reaction rates. Kinetics experiments require reactants to be mixed on a time scale faster than that of the reaction. Unfortunately, highly concentrated solutions of crowding agents such as bovine serum albumin and hemoglobin that are viscous and sticky are extremely difficult to mix rapidly. We demonstrate a new droplet-based microfluidic mixer that induces chaotic mixing of crowded solutions in milliseconds due to protrusions of the microchannel walls that generate oscillating interfacial shear within the droplets. Mixing in the microfluidic mixer is characterized, mechanisms underlying mixing are discussed, and evidence of biocompatibility is presented. This microfluidic platform will allow for the first kinetic studies of biological reactions with millisecond time resolution under conditions of macromolecular crowding similar to those within cells. PMID- 16316170 TI - Combination of sustained off-resonance irradiation and on-resonance excitation in FT-ICR. AB - Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry is becoming more widely used among the mass spectrometric techniques and has excellent figures of merit. Ion activation and fragmentation via sustained off-resonance irradiation (SORI) collision-induced dissociation (CID) is commonly used in FT ICR. However, one of the limitations of SORI-CID is that only low-energy processes are typically observed in the product ion spectra. Here we present another option for performing CID in FT-ICR, a combination of SORI and on resonance excitation (RE), termed SORI-RE. In comparison to SORI, this method produces more abundant ions resulting from higher energy fragmentation pathways. The result is the observation of a significant abundance of both higher and lower energy fragmentation pathways in the same mass spectrum. The comparison of SORI, RE, and SORI-RE spectra may lead to mechanistic insights as the relative abundances of certain fragment ions change as a function of internal energy deposition. This technique is simple to incorporate in existing instruments, does not require hardware or software modification, and requires only an additional 20 40 ms acquisition time. The technique is illustrated for a peptide (YGGFL), two disaccharides differing in the position of the glycosidic linkage (2alpha mannobiose, 3alpha-mannobiose), an oligosaccharide (Alditol XT), a small protein (ubiquitin), and an inorganic cation (UO2+). Examples of higher energy fragmentation pathways enhanced by SORI-RE include the formation of immonium ions and oligosaccharide cross-ring cleavages. PMID- 16316171 TI - Analysis of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl) 1-butanol in urine by extraction on a molecularly imprinted polymer column and liquid chromatography/atmospheric pressure ionization tandem mass spectrometry. AB - The tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) is present in the urine of tobacco users and, at lower concentrations, in the urine of nonsmokers exposed to secondhand smoke. NNAL is a valuable biomarker of human exposure to the carcinogenic nitrosamines in tobacco and tobacco smoke, but its presence at low concentrations in urine requires sensitive and often complex analytic procedures. In this report, we describe the development of an efficient method for the analysis of NNAL in human urine using liquid chromatography/atmospheric pressure ionization tandem mass spectrometry (LC/MS/MS) combined with a novel sample cleanup based on a molecularly imprinted polymer (MIP) column developed specifically for this assay. Our results suggest that this combination of MIP column extraction and LC/MS/MS can provide a sensitive and relatively simple analytical method suitable for application to epidemiologic investigations of health risks associated with the exposure to tobacco smoke or SHS in both smokers and nonsmokers. PMID- 16316172 TI - Measurement of the isotope enrichment of stable isotope-labeled proteins using high-resolution mass spectra of peptides. AB - Stable isotope-enriched molecules are used as internal standards and as tracers of in vivo substrate metabolism. The accurate conversion of measured ratios in the mass spectrometer to mole ratios is complicated because a polyatomic molecule containing enriched atoms will result in a combinatorial distribution of isotopomers depending on the enrichment and number of "labeled" atoms. This effect could potentially cause a large error in the mole ratio measurement depending on which isotope peak or peaks were used to determine the ratio. We report a computational method that predicts isotope distributions over a range of enrichments and compares the predicted distributions to experimental peptide isotope distributions obtained by Fourier transform ion cyclotron resonance mass spectrometry. Our approach is accurate with measured enrichments within 1.5% of expected isotope distributions. The method is also precise with 4.9, 2.0, and 0.8% relative standard deviations for peptides containing 59, 79, and 99 atom % excess (15)N, respectively. The approach is automated making isotope enrichment calculations possible for thousands of peptides in a single muLC-FTICR-MS experiment. PMID- 16316173 TI - Determination of endocrine disrupting compounds in marine water by nanoliquid chromatography/direct-electron ionization mass spectrometry. AB - We present a new method for the determination of 29 endocrine-disrupting compounds in marine water. This method is based on a solid-phase extraction preconcentration technique, followed by a nanoscale liquid chromatography/direct electron ionization (EI) mass spectrometric analysis. Direct-EI is a novel technique for the rapid conversion of a GC/MS into an efficient and reliable LC/MS for EI detection. The capability to acquire EI mass spectra of the analytes, and to operate in selected ion monitoring mode during real sample analyses, allows certain identification and precise quantification. In addition, this method is not influenced by the polarity of the analytes and does not require different detection modes (positive and negative) for identification with API techniques. Limits of detection of the method span from 0.4 to 118.7 ng.L( 1), corresponding to an instrumental detection limit of 0.005-1.260 ng. Linear regression and recovery experiment data, together with their standard deviations, are also presented. Marine water samples were collected along the middle-western Adriatic Coast (Italy), near the shore and at the mouth of rivers and canals. PMID- 16316174 TI - Potential of gas chromatography coupled to triple quadrupole mass spectrometry for quantification and confirmation of organohalogen xenoestrogen compounds in human breast tissues. AB - The potential of gas chromatography coupled to tandem mass spectrometry (GC/MS/MS) with a triple quadrupole analyzer (QqQ) has been investigated for the accurate and sensitive determination of xenoestrogens in human breast tissues. Special emphasis has been given to the confirmation of the identity of compounds detected in the samples analyzed in order to avoid the reporting of false positives. The work has been focused on the determination of approximately 30 organochlorine compounds (PCBs and pesticides) and organobromine compounds (polybrominated diphenyl ethers) in adipose breast tissue and in tumoral fragment. Analytes were extracted by dissolving the samples in hexane, and the extracts were purified by automated normal-phase HPLC prior to GC/MS/MS analysis. Three isotopically labeled standards were added before extraction as surrogates for the quality control of the analyses. Accuracy and precision were evaluated by means of recovery experiments using adipose breast tissue spiked at three concentration levels, with satisfactory results for most analytes. The excellent selectivity and sensitivity of QqQ in selected reaction monitoring mode allowed us satisfactory quantification and confirmation at levels as low as 5-25 ng/g, i.e., the lowest concentration level for which the method was fully validated. Two MS/MS transitions were selected for each analyte, using the concentration ratio obtained from them as a confirmatory parameter. The developed methodology was applied to the analysis of 51 breast samples (26 adipose tissues and 25 tumoral fragments), giving as a result the detection and confirmation of several organochlorine compounds in both types of samples. Due to its adequate analytical characteristics, the optimized method fits with the requirements of accurate quantification and reliable confirmation of the identity of compounds detected according to the most recent European Guidelines. As an ultimate unequivocal confirmation, several selected samples were reanalyzed by gas chromatography coupled to mass spectrometry with a time-of-flight (TOF) analyzer. Confirmation of analytes present at higher concentrations was successful with mass error less than 5 mDa. However, confirmation by TOF MS was not possible al low concentrations (i.e., at the few ng/g level) as a consequence of its lower sensitivity compared with that of triple quadrupole in selected reaction monitoring mode. PMID- 16316175 TI - Multiplexed flow cytometric immunoassay for influenza virus detection and differentiation. AB - Microsphere-based immunoassay by flow cytometry has gained popularity lately in protein detection and infectious disease diagnosis due to its capacity for multiplexed analysis and simple assay format. Here, we demonstrated the power of microsphere-based immunoassay for high-sensitivity detection and accurate differentiation of influenza viruses. The effects of sample volume and bead number on the assay sensitivity of viral antigen detection were studied. Compared to enzyme-linked immunosorbent assays, flow-based bead assays provided approximately 10-fold lower detection limit for viral particle detection and performed similarly for recombinant viral hemagglutinin protein detection. A four plexed assay for influenza virus typing and influenza B virus sublineage characterization was developed to demonstrate the potential for multiplexed viral antigen detection and differentiation. PMID- 16316176 TI - Photocatalytic degradation-excitation-emission matrix fluorescence for increasing the selectivity of polycyclic aromatic hydrocarbon analyses. AB - The application of photocatalysis enhancement to calibration of fluorescence excitation-emission matrixes (EEMs) with parallel factor (PARAFAC) analysis is described. In this study, three- and four-way PARAFAC analysis was employed to extract the fluorescent species' spectra from overlapping EEMs. Time-dependent photocatalysis degradation of the polycyclic aromatic hydrocarbons (PAHs) was employed to create an additional dimension for analysis. The consequent four dimension degradation-EEM data cubes have greater selectivity for each PAH than do three-dimension EEM data cubes alone. On a scale of 0 to 1, with 0 being completely collinear spectra and 1 being orthogonal spectra, including the time dependent measurements increased the selectivity an average of 21%, from 0.73 to 0.87. PMID- 16316177 TI - Antioxidant sensors based on DNA-modified electrodes. AB - TiO2/ITO modified electrodes were developed to quantitatively photooxidize adsorbed ds-DNA and to study the effect of antioxidants as ds-DNA protecting agents. TiO2 films are used for efficient ds-DNA immobilization, for ds-DNA oxidation through photogenerated hydroxyl radicals, and as electrodes for amperometric sensing. The films, prepared by a sol-gel process, are deposited on ITO glass electrodes. Damages occurring after ds-DNA oxidation by ROS are detected by adding MB as an intercalant probe and by monitoring the electrochemical reduction current of the intercalated redox probe. The MB electrochemical signal is found to be sensitive enough to monitor ds-DNA structure changes, and the electrochemical sensor has been applied to the evaluation of the antioxidant properties of glutathione and gallic acid. PMID- 16316178 TI - pH-sensitive solid-state electrode based on electrodeposited nanoporous platinum. AB - The nanoporous platinum oxide (H1-ePtO) as a hydrogen ion-selective sensing material is reported. Bare nanoporous platinum oxides exhibit near-Nernstian behavior (e.g., -55 mV/pH in PBS), ignorable hysteresis, a short response time, and high precision, which are remarkably better than those of flat platinum oxides. The electrode potential of a nanoporous platinum oxide responds exclusively to hydrogen ion, which implies its usefulness as a solid-state pH sensor. In the present study, the performance of nanoporous platinum oxide was investigated and compared with that of IrOx in terms of selectivity and the influences of ionic strength, temperature, complexing ligands, and surfactants. H1-ePtO functions well as a pH-sensing solid-state material, and it is viewed as a promising alternative to IrOx. Interference by redox couples was successfully suppressed by covering the H1-ePtO surface with a protective layer, e.g., an electropolymerized polyphenol thin film. Since the nanoporous platinum oxide with such a protective layer is particularly suitable for miniaturization and micropatterning, our findings suggest its usefulness in applications such as solid-state pH sensors embedded in chip-based microanalysis systems. PMID- 16316179 TI - Microfluidic electrophoresis chip coupled to microdialysis for in vivo monitoring of amino acid neurotransmitters. AB - Microfluidic electrophoresis devices were coupled on-line to microdialysis for in vivo monitoring of primary amine neurotransmitters in rat brain. The devices contained a sample introduction channel for dialysate, a precolumn reactor for derivatization with o-phthaldialdehyde, a flow-gated injector, and a separation channel. Detection was performed using confocal laser-induced fluorescence. In vitro testing revealed that the initial device design had detection limits for amino acids of approximately 200 nM, relative standard deviation of peak heights of 2%, and separations within 95 s with up to 30,200 theoretical plates when applying an electric field of 370 V/cm. A second device design that allowed electric fields of 1320 V/cm to be applied while preserving the reaction time allowed separations within 20 s with up to 156,000 theoretical plates. Flow splitting into the electrokinetic network from hydrodynamic flow in the sample introduction channel was made negligible for sampling flow rates from 0.3 to 1.2 microL/min by placing a 360-microm-diameter fluidic access hole that had flow resistance (0.15-7.2) x 10(8)-fold lower than that of the electrokinetic network at the junction of the sample introduction channel and the electrokinetic network. Using serial injections, the device allowed the dialysate stream to be analyzed at 130-s intervals. In vivo monitoring was demonstrated by using the microdialysis/microfluidic device to record glutamate concentrations in the striatum of an anesthetized rat during infusion of the glutamate uptake inhibitor l-trans-pyrrolidine-2,4-dicarboxylic acid. These results prove the feasibility of using a microfabricated fluidic system coupled to sampling probes for chemical monitoring of complex media such as mammalian brain. PMID- 16316180 TI - Combining peptide modeling and capillary electrophoresis-mass spectrometry for characterization of enzymes cleavage patterns: recombinant versus natural bovine pepsin A. AB - Nowadays there is an increasing number of recombinant enzymes made available to industry. Before replacing the use of natural enzymes with their cognate recombinant counterparts, one important issue to address is their actual equivalence. For a given recombinant proteolytic enzyme, its equivalence can be investigated by comparing its cleavage specificity with that obtained from the natural enzyme. This is mostly done by analyzing the fragments (i.e., peptidic map) attained after enzymatic digestion of a given protein used as substrate. The peptidic maps obtained are typically characterized using separation techniques together with MS and MS/MS systems. However, these procedures are known to be difficult and labor-intensive. In this work, the combined use of a theoretical model that relates electrophoretic behavior of peptides to their sequence together with capillary electrophoresis-mass spectrometry (CE-MS) is proposed to characterize in a very fast and simple way the cleavage specificity of new recombinant enzymes. Namely, the effectiveness of this procedure is demonstrated by analyzing in few minutes the fragments obtained from a protein hydrolysated using recombinant and natural pepsin A. The usefulness of this strategy is further corroborated by CE-MS/MS. The proposed procedure is applicable in many other proteomic studies involving CE-MS of peptides. PMID- 16316181 TI - Use of semiselective TOCSY and the pearson correlation for the metabonomic analysis of biofluid mixtures: application to urine. AB - The authors recently proposed an approach to the metabonomic analysis of biofluid mixtures based on the use of the selective TOCSY experiment (Sandusky, P.; Raftery, D. Anal. Chem. 2005, 77, 2455). This method has some significant advantages over standard metabonomic analysis. However, when analyzing overlapped components, the selective TOCSY method can suffer from the relatively high likelihood of simultaneous excitation of several spin systems at once. This multiple excitation can cause problems both with the purity of the individual TOCSY peaks observed and with their assignment into specific spin systems. To address this problem, the possibility of using a more selective excitation is initially explored. Unfortunately, in most cases, greater spin system selectivity can only be gained at the expense of sensitivity. This is obviously an unacceptable tradeoff when dealing with biofluid samples. However, the application of the Pearson product moment correlation to the TOCSY peak integral intensities provides a test for individual TOCSY peak purity and allows for the assignment of the peaks into spin systems. The specific application of this two stage "semiselective" TOCSY method to rat and human urine is presented. Significantly, it is also demonstrated that the use of semiselective TOCSY spectra as data inputs for PCA calculations provides a more sensitive and reliable method of distinguishing small differences in biofluid composition than the standard metabonomic approach using complete 1D proton NMR spectra of urine samples. PMID- 16316182 TI - Species-specific isotope dilution analysis and isotope pattern deconvolution for butyltin compounds metabolism investigations. AB - A methodology for the study of the absorption and metabolism of butyltin compounds in laboratory animals using isotopically enriched species was developed. The method is based on the oral administration of 119Sn-labeled monobutyltin (MBT), 118Sn-labeled dibutyltin (DBT), and 117Sn-labeled tributyltin (TBT) to the animals and the measurement of both the concentration and isotopic composition of these compounds in the different tissues by GC-ICPMS. The degradation of butyltin compounds during their metabolism was computed using least-squares isotope pattern deconvolution, and their concentration was measured by reverse isotope dilution analysis using natural-abundance MBT, DBT, and TBT standards. Male Wistar rats were used as models to evaluate the proposed methodology. Preliminary toxicological results obtained with one rat indicate that TBT is highly absorbed (64.4%), and it is found in all organs with relatively high levels in stomach and intestines. The apparent absorption of DBT was 27.3% and was mainly found in liver, kidney, and intestines. However, a large proportion of the found DBT is formed from the degradation of TBT (approximately 40% of the found DBT in liver is degraded TBT). The apparent absorption of MBT was found to be 12.5%, and the originally administered MBT was mainly recovered in the feces. However, MBT was clearly detected in liver, kidney, stomach, intestines, and urine as degradation products of DBT and TBT. Although a significant variability from rat to rat is expected to be obtained, the analytical variability provided by this methodology is small enough to yield meaningful biological results. The results obtained demonstrate that the developed methodology is able to follow qualitatively, quantitatively, and simultaneously the specific metabolic pathways of different species of a given element. PMID- 16316183 TI - A comprehensive two-dimensional retention time alignment algorithm to enhance chemometric analysis of comprehensive two-dimensional separation data. AB - A comprehensive two-dimensional (2D) retention time alignment algorithm was developed using a novel indexing scheme. The algorithm is termed comprehensive because it functions to correct the entire chromatogram in both dimensions and it preserves the separation information in both dimensions. Although the algorithm is demonstrated by correcting comprehensive two-dimensional gas chromatography (GC x GC) data, the algorithm is designed to correct shifting in all forms of 2D separations, such as LC x LC, LC x CE, CE x CE, and LC x GC. This 2D alignment algorithm was applied to three different data sets composed of replicate GC x GC separations of (1) three 22-component control mixtures, (2) three gasoline samples, and (3) three diesel samples. The three data sets were collected using slightly different temperature or pressure programs to engender significant retention time shifting in the raw data and then demonstrate subsequent corrections of that shifting upon comprehensive 2D alignment of the data sets. Thirty 12-min GC x GC separations from three 22-component control mixtures were used to evaluate the 2D alignment performance (10 runs/mixture). The average standard deviation of first column retention time improved 5-fold from 0.020 min (before alignment) to 0.004 min (after alignment). Concurrently, the average standard deviation of second column retention time improved 4-fold from 3.5 ms (before alignment) to 0.8 ms (after alignment). Alignment of the 30 control mixture chromatograms took 20 min. The quantitative integrity of the GC x GC data following 2D alignment was also investigated. The mean integrated signal was determined for all components in the three 22-component mixtures for all 30 replicates. The average percent difference in the integrated signal for each component before and after alignment was 2.6%. Singular value decomposition (SVD) was applied to the 22-component control mixture data before and after alignment to show the restoration of trilinearity to the data, since trilinearity benefits chemometric analysis. By applying comprehensive 2D retention time alignment to all three data sets (control mixtures, gasoline samples, and diesel samples), classification by principal component analysis (PCA) substantially improved, resulting in 100% accurate scores clustering. PMID- 16316184 TI - Characterization of synthetic polymers by electrospray-assisted pyrolysis ionization-mass spectrometry. AB - Rapid characterization of synthetic polymers based on the differences in the appearance of the polar pyrolysate ions was achieved by electrospray-assisted pyrolysis ionization-mass spectrometry. The pyrolytical products produced by a commercial Curie point pyroprobe were conducted to the tip of a capillary, where charged methanol droplets were generated continuously by electrospray. Polar components in the pyrolysates may react directly with the protonated methanol ions or fuse with the charged methanol droplets; electrospray ionization proceeds from the fused droplets to generate protonated analyte ions. The mass spectra obtained through this approach were used to rapidly distinguish the polymer standards that differ in the nature of building units, degrees of polymerization, and copolymerization coefficients. PMID- 16316185 TI - Dioxin-binding pentapeptide for use in a high-sensitivity on-bead detection assay. AB - The purpose of this study is to develop a dioxin detection method using a short peptide alternative to an immunoantibody. A full peptide library consisting of 2.5 million possible amino acid combinations was constructed by a solid-phase split synthesis approach using 19 natural amino acids. The peptide beads were subjected to a competitive binding assay between 2,3,7-trichlorodibenzo-p-dioxin and N-NBD-3-(3',4'-dichlorophenoxy)-1-propylamine (NBD-DCPPA) in a buffer containing 20% 1,4-dioxane. Two almost identical pentapeptides, FLDQI and FLDQV, that could bind dioxin were screened from the combinatorial library. NBD-DCPPA and the peptide synthesized on resin beads could be utilized to determine dioxin concentrations. The fluorescence intensity of the beads was measured using fluorescence microscopy to make a calibration curve for the dioxin concentrations. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TeCDD) could also detected in the presence of 30% 1,4-dioxane. To optimize the peptide sequence, a one-amino acid-substituted library was prepared using amino acids including nonnatural amino acids. The internal amino acids, LDQ, could not be substituted by any other amino acids. This result indicates that these three side chains are essential to recognize dioxins. The peptide C terminus substituted by phenylglycine showed a 10 times lower detection limit of 2,3,7,8-TeCDD of 150 pM (50 pg/mL) than the original sequence FLDQV. The cross reactivity of the dioxin binding peptides including the secondary derivatives was investigated. Some polycyclic aromatic hydrocarbons bound to the peptide beads, but nonchlorinated dibenzo-p-dioxin and PCB did not. From these results, we demonstrate the potential of short peptides as a practical sensor material targeting low molecular weight compounds such as dioxin. PMID- 16316186 TI - Reduction of the nonspecific binding of a target antibody and of its enzyme labeled detection probe enabling electrochemical immunoassay of an antibody through the 7 pg/ml-100 ng/mL (40 fM-400 pM) range. AB - We describe a simple, potentially low-cost, amperometric, enzyme-amplified, sandwich-type immunoassay, monitoring IgG at a concentration as low as approximately 7 pg/mL with a dynamic range of 10(4). The assay utilizes a screen printed carbon electrode on which a redox hydrogel and avidin are co electrodeposited. To neutralize nonspecifically binding positively charged microdomains of the avidin, two polyanions, poly(acrylic acid-co-maleic acid) and poly(acrylic acid), are applied. These polyanions bind to the film not only electrostatically but also by Michael addition reaction to cysteine, lysine, or arginine functions of the avidin. The electrode is then made specific for the analyte, for which rabbit IgG was chosen, by conjugating the film-bound avidin to biotin-labeled anti-rabbit IgG. After exposure to the tested solution and capture of rabbit IgG, the sandwich is completed by conjugation of horseradish-peroxidase (HRP)-labeled anti-rabbit IgG. Electrical contact between the HRP and the electrode-bound hydrogel results in the formation of an electrocatalyst for the electroreduction of H2O2 to water. The application of the poly(acrylic acid-co maleic acid) and the poly(acrylic acid) reduces the nonspecific adsorption associated noise, lowers the detection limit from 3 ng/mL (approximately 20 pM analyte antibody concentration) to approximately 7 pg/mL (approximately 40 fM analyte antibody concentration), and also expands the dynamic range to 10(4). PMID- 16316187 TI - Making broad proteome protein measurements in 1-5 min using high-speed RPLC separations and high-accuracy mass measurements. AB - The throughput of proteomics measurements that provide broad protein coverage is limited by the quality and speed of both the separations as well as the subsequent mass spectrometric analysis; at present, analysis times can range anywhere from hours (high throughput) to days or longer (low throughput). We have explored the basis for proteomics analyses conducted on the order of minutes using high-speed capillary RPLC combined through on-line electrospray ionization interface with high-accuracy mass spectrometry (MS) measurements. Short 0.8 microm porous C18 particle-packed 50-microm-i.d. capillaries were used to speed the RPLC separations while still providing high-quality separations. Both time-of flight (TOF) and Fourier transform ion cyclotron resonance (FTICR) MS were applied for identifying peptides using the accurate mass and time (AMT) tag approach. Peptide RPLC relative retention (elution) times that were generated by solvent gradients that differed by at least 25-fold were found to provide relative elution times that agreed to within 5%, which provides the basis for using peptide AMT tags for higher throughput proteomics measurements. For fast MS acquisition speeds (e.g., 0.2 s for TOF and either approximately 0.3 or approximately 0.6 s for FTICR), peptide mass measurement accuracies of better than +/-15 ppm were obtained with the high-speed RPLC separations. The ability to identify peptides and the overall proteome coverage was determined by factors that include the separation peak capacity, the sensitivity of the MS (with fast scanning), and the accuracy of both the mass measurements and the relative RPLC peptide elution times. The experimental RPLC relative elution time accuracies of 5% (using high-speed capillary RPLC) and mass measurement accuracies of better than +/-15 ppm allowed for the confident identification of >2800 peptides and >760 proteins from >13,000 different putative peptides detected from a Shewanellaoneidensis tryptic digest. Initial results for both RPLC-ESI-TOF and RPLC-ESI-FTICR MS were similar, with approximately 2000 different peptides from approximately 600 different proteins identified within 2-3 min. For <120-s proteomic analysis, TOF MS analyses were more effective, while FTICR MS was more effective for the >150-s analysis due to the improved mass accuracies attained using longer spectrum acquisition times. PMID- 16316188 TI - Identification of unusual bacterial glycosylation by tandem mass spectrometry analyses of intact proteins. AB - The characterization of protein glycosylation can be a complex and time-consuming procedure, especially for prokaryote O-linked glycoproteins, which often comprise unusual oligosaccharide structures with no known glycosylation motif. In this report, we describe a "top-down" approach that provides information on the extent of glycosylation, the molecular masses, and the structure of oligosaccharide residues on bacterial flagella, important structural proteins involved in the motility of pathogenic bacteria. Flagella from four bacterial pathogens, namely, Campylobacter jejuni, Helicobacter pylori, Aeromonas caviae, and Listeria monocytogenes, were analyzed by this top-down mass spectrometry approach. The approach needs minimal sample preparation and can be performed within a few minutes compared to the tedious and often time-consuming "bottom-up" approach involving proteolytic digestion and LC-MS-MS analyses of the suspected glycopeptides. Multiply protonated protein precursor ions subjected to low-energy collisional activation in a quadrupole time-of-flight instrument showed extensive and specific gas-phase deglycosylation resulting in the formation of abundant oxonium ions with very few fragment ions from peptidic bond cleavages. Structural information on individual carbohydrate residues is obtained using a second generation product ion scan of oxonium ions formed by collisional activation of the intact protein ions in the source region. The four bacterial flagella examined differed not only by the extent of glycosylation but also by the nature of carbohydrate substituents. For example, the flagellin from the Gram-positive bacterium, L. monocytogenes showed O-linked GlcNAc residues at up to 6 sites/protein monomer. In contrast, the three Gram-negative bacterial pathogens C. jejuni, H. pylori and A. caviae displayed up to 19 Ser/Thr O-linked sites modified with residues structurally related to N-acetylpseudaminic acid (Pse5Ac7Ac) and in the case of Campylobacter include a novel N-acetylglutamine substituent on Pse5Am7Ac. PMID- 16316189 TI - De novo sequencing of neuropeptides using reductive isotopic methylation and investigation of ESI QTOF MS/MS fragmentation pattern of neuropeptides with N terminal dimethylation. AB - A stable-isotope dimethyl labeling strategy was previously shown to be a useful tool for quantitative proteomics. More recently, N-terminal dimethyl labeling was also reported for peptide sequencing in combination with database searching. Here, we extend these previous studies by incorporating N-terminal isotopic dimethylation for de novo sequencing of neuropeptides directly from tissue extract without any genomic information. We demonstrated several new sequencing applications of this method in addition to the identification of the N-terminal residue using the enhanced a(1) ion. The isotopic labeling also provides easier and more confident de novo sequencing of peptides by comparing similar MS/MS fragmentation patterns of the isotopically labeled peptide pairs. The current study on neuropeptides shows several distinct fragmentation patterns after N terminal dimethylation which have not been reported previously. The y((n-1)) ion is enhanced in multiply charged peptides and is weak or missing in singly charged peptides. The MS/MS spectra of singly charged peptides are simplified due to the enhanced N-terminal fragments and suppressed internal fragments. The neutral loss of dimethylamine is also observed. The mechanisms for the above fragmentations are proposed. Finally, the structures of the immonium ion and related ions of N(alpha), N(epsilon)-tetramethylated lysine and N(epsilon)-dimethylated lysine are explored. PMID- 16316190 TI - Development of mass spectrometric method for analysis of cyclic nitramine explosives DTIW and HNIW. AB - Mass spectrometric (MS) methods are used for the analysis of two novel nitramine explosives-hexanitrohexaazaisowurzitane (HNIW) and 4,10-dinitro-2,6,8,12-tetraoxa 4,10-diazaisowurzitane (DTIW). The methods include electrospray (ESI) and atmospheric pressure chemical ionization techniques for liquid chromatography/MS (LC/MS), chemical ionization for direct introduction (DCI), and gas chromatography/MS (CI-GC/MS). It is found that HNIW (438 Da) is detectable using both positive and negative modes of DCI and in the negative mode ESI-MS. Several anions were found to complex with HNIW, e.g., CF3CO2-, Cl-, Br-, I-, NO3-, and NO2-. On the other hand, DTIW could only be detected using positive DCI and CI GC/MS, where an MH+ ion (m/z 263) was formed. The fragmentation pathways of the two nitramines were further studied by MS2 experiments. Apparently, the main fragmentation pathway of the MH+ ion of DTIW involves the loss of nitrous acid. Several anion adducts of HNIW that were studied dissociate to afford neutral HNIW and the added anions. However, Cl-, Br-, I-, and NO2- afford a series of fragments that resulted from the dissociation of the isowurzitane structure. For these anions, limit of detection was also found. To understand some of the HNIW fragmentation pathways, DFT calculations were used. PMID- 16316191 TI - Ion-selective supported liquid membranes placed under steady-state diffusion control. AB - Supported liquid membranes are used here to establish steady-state concentration profiles across ion-selective membranes rapidly and reproducibly. This opens up new avenues in the area of nonequilibrium potentiometry, where reproducible accumulation and depletion processes at ion-selective membranes may be used to gain valuable analytical information about the sample. Until today, drifting signals originating from a slowly developing concentration profile across the ion selective membrane made such approaches impractical in zero current potentiometry. Here, calcium- and silver-selective membranes were placed between two identical aqueous electrolyte solutions, and the open circuit potential was monitored upon changing the composition of one solution. Steady state was reached in approximately 1 min with 25-microm porous polypropylene membranes filled with bis(2-ethylhexyl) sebacate doped with ionophore and lipophilic ion exchanger. Ion transport across the membrane resulted on the basis of nonsymmetric ion-exchange processes at both membrane sides. The steady-state potential was calculated as the sum of the two membrane phase boundary potentials, and good correspondence to experiment was observed. Concentration polarizations in the contacting aqueous phases were confirmed with stirring experiments. It was found that interferences (barium in the case of calcium electrodes and potassium with silver electrodes) induce a larger potential change than expected with the Nicolsky equation because they influence the level of polarization of the primary ion (calcium or silver) that remains potential determining. PMID- 16316192 TI - Detection of human immunodeficiency virus type 1 DNA sequence using plasmonics nanoprobes. AB - This paper describes the use of plasmonics-based nanoprobes that act as molecular sentinels for DNA diagnostics. The plasmonics nanoprobe comprises a metal nanoparticle and a stem-loop DNA molecule tagged with a Raman label. The nanoprobe utilizes the specificity and selectivity of the DNA hairpin probe sequence to detect a specific target DNA sequence of interest. In the absence of target DNA, the stem-loop configuration maintains the Raman label in proximity to the metal nanoparticle, inducing an intense surface-enhanced Raman scattering (SERS) effect that produces a strong Raman signal upon laser excitation. Upon hybridization of a complementary target DNA sequence to the nanoprobe, the stem loop configuration is disrupted, causing the Raman label to physically separate from the metal nanoparticle, thus quenching the SERS signal. The usefulness and potential application of the plasmonics nanoprobe for diagnosis is demonstrated using the gag gene sequence of the human immunodeficiency virus type 1 (HIV-1). We successfully demonstrated the specificity and selectivity of the plasmonics nanoprobes to detect PCR amplicons of the HIV gene. The potential for combining the spectral selectivity and high sensitivity of the SERS process with inherent molecular specificity of DNA hairpins to diagnose molecular target sequences in homogeneous solutions is discussed. PMID- 16316193 TI - Enhanced characterization of complex proteomic samples using LC-MALDI MS/MS: exclusion of redundant peptides from MS/MS analysis in replicate runs. AB - Due to the complexity of proteome samples, only a portion of peptides and thus proteins can be identified in a single LC-MS/MS analysis in current shotgun proteomics methodologies. It has been shown that replicate runs can be used to improve the comprehensiveness of the proteome analysis; however, high-intensity peptides tend to be analyzed repeatedly in different runs, thus reducing the chance of identifying low-intensity peptides. In contrast to commonly used online ESI-MS, offline MALDI decouples the separation from MS acquisition, thus allowing in-depth selection for specific precursor ions. Accordingly, we extended a strategy for offline LC-MALDI MS/MS analysis using a precursor ion exclusion list consisting of all identified peptides in preceding runs. The exclusion list eliminated redundant MS/MS acquisitions in subsequent runs, thus reducing MALDI sample depletion and allowing identification of a larger number of peptide identifications in the cumulative dataset. In the analysis of the digest of an Escherichia coli lysate, the exclusion list strategy resulted in a 25% increase in the number of unique peptide identifications in the second run, in contrast to simply pooling MS/MS data from two replicate runs. To reduce the increased LC analysis time for repeat runs, a four-column multiplexed LC system was developed to carry out separation simultaneously. The multiplexed LC-MALDI MS provides a high-throughput platform to utilize the exclusion list strategy in proteome analysis. PMID- 16316194 TI - Analysis of solids, liquids, and biological tissues using solids probe introduction at atmospheric pressure on commercial LC/MS instruments. AB - Direct analysis of samples using atmospheric pressure ionization (API) provides a more rapid method for analysis of volatile and semivolatile compounds than vacuum solids probe methods and can be accomplished on commercial API mass spectrometers. With only a simple modification to either an electrospray (ESI) or atmospheric pressure chemical ionization (APCI) source, solid as well as liquid samples can be analyzed in seconds. The method acts as a fast solids/liquid probe introduction as well as an alternative to the new direct analysis in real time (DART) and desorption electrospray ionization (DESI) methods for many compound types. Vaporization of materials occurs in the hot nitrogen gas stream flowing from an ESI or APCI probe. Ionization of the thermally induced vapors occurs by corona discharge under standard APCI conditions. Accurate mass and mass-selected fragmentation are demonstrated as is the ability to obtain ions from biological tissue, currency, and other objects placed in the path of the hot nitrogen stream. PMID- 16316195 TI - Fabricating RNA microarrays with RNA-DNA surface ligation chemistry. AB - A novel surface attachment strategy that utilizes RNA-DNA surface ligation chemistry to create renewable RNA microarrays from single-stranded DNA (ssDNA) microarrays on gold surfaces is demonstrated. The enzyme T4 DNA ligase was used to catalyze the formation of a phosphodiester bond between 5'-phosphate-modified ssDNA attached to the surface and the 3'-hydroxyl group of unlabeled RNA molecules from solution in the presence of a complementary template DNA strand. Surface plasmon resonance imaging (SPRI) measurements were performed to characterize the ligation process as well as to verify the bioactivity of the ssRNA microarray in terms of (i) the hybridization adsorption of complementary DNA onto the RNA array to form a surface RNA-DNA heteroduplex and (ii) the hydrolysis of the RNA microarrays with either ribonuclease S or ribonuclease H (RNase H). The hydrolysis of the surface-bound RNA with RNase H required the presence of a surface heteroduplex and, upon completion, regenerated the original 5'-phosphate-terminated ssDNA array elements. These ssDNA array elements could be ligated again to create a new RNA microarray. These RNA microarrays can be used in the study of RNA-protein/RNA/aptamer bioaffinity interactions and for the enzymatically amplified SPRI detection of DNA in the presence of RNase H. PMID- 16316196 TI - Perchlorate isotope forensics. AB - Perchlorate has been detected recently in a variety of soils, waters, plants, and food products at levels that may be detrimental to human health. These discoveries have generated considerable interest in perchlorate source identification. In this study, comprehensive stable isotope analyses (37Cl/35Cl and 18O/17O/16O) of perchlorate from known synthetic and natural sources reveal systematic differences in isotopic characteristics that are related to the formation mechanisms. In addition, isotopic analyses of perchlorate extracted from groundwater and surface water demonstrate the feasibility of identifying perchlorate sources in contaminated environments on the basis of this technique. Both natural and synthetic sources of perchlorate have been identified in water samples from some perchlorate occurrences in the United States by the isotopic method. PMID- 16316197 TI - Two generalizable routes to terminal carbido complexes. AB - Desulfurization of the thiocarbonyl ligand in square pyramidal [Ru(CS)Cl2(PCy3)2] (1-S) via sulfur atom abstraction using [Mo(H)(eta2-Me2CNAr)(N[i-Pr]Ar)2] forms [Ru(C)Cl2(PCy3)2] (1) cleanly over several hours in benzene; isolated yield is 55%. Complex 1 is also formed in 87% isolated yield upon reaction of [Ru(CHR)(PCy3)2Cl2] (R = p-C6H4Me, 2; Ph, 3) with vinyl acetate in dichloromethane. Complex 1-S is re-formed quantitatively from 1 upon treatment with elemental sulfur in CH2Cl2, but is prepared most conveniently by treatment of crude [Ru(CS)Cl2(PPh3)2(OH2)] with excess PCy3 in toluene. Nearly quantitative conversion of 1 to [Ru(CO)Cl2(PCy3)2] (1-O) occurs upon addition of dimethyldioxirane solution in acetone to 1 dissolved in CH2Cl2 at ca. -90 degrees C. PMID- 16316198 TI - Incorporating lanthanide cations with cadmium selenide nanocrystals: a strategy to sensitize and protect Tb(III). AB - The electronic structure of CdSe semiconductor nanocrystals has been used to sensitize Tb3+ in solution by incorporation of Tb3+ cations into the nanocrystals during synthesis. Doping of luminescent Tb3+ metal ions in semiconductor nanocrystals utilizes the positive attributes of both species' photophysical properties, resulting in a final product with long luminescence lifetimes, sharp emission bands, high absorptivities, and strong resistance to decomposition. This strategy also helps protect the lanthanide cations from nonradiative deactivation from C-H, N-H, and O-H oscillators of solvent molecules or traditional organic lanthanide ligands, leading to long Tb3+ luminescence lifetimes. This new type of nanomaterial synergistically combines the photophysical properties of nanocrystals and Tb3+. PMID- 16316199 TI - Synthesis, structure, and reactivity of unexpectedly stable spiroepoxy-beta lactones obtained by epoxidation of 4-alkylidene-2-oxetanones. AB - We describe the first synthesis of spiroepoxy-beta-lactones obtained via epoxidation of ketene dimers. These compounds display unexpected stability that may be due to a double anomeric effect garnered from analysis of bond lengths by X-ray crystallography of one spirocycle in comparison to calculated bond lengths of related structures. These new strained intermediates display interesting reactivity leading to a butenolide, an alpha-hydroxyketone, a triol, an alpha chloroketone, and an alpha-azidoketone. PMID- 16316200 TI - In situ fabrication of three-dimensional chemical patterns in fused silica separation capillaries with polymerized phospholipids. AB - We report a new molecular approach for in situ generation of micron scale, chemically and biochemically functionalized patterns inside three-dimensional, completely enclosed fluidic channels. The formation of chemical patterns is based upon a combination of lipid bilayer self-assembly and UV photopolymerization of photoreactive, cross-linkable phospholipids. Using this approach, we have functionalized capillaries of varying inner diameters with a range of chemistries useful for protein and peptide immobilization. Here, we demonstrate the ability to produce small molecule and protein-based chemical patterns. PMID- 16316201 TI - Air-breathing laminar flow-based microfluidic fuel cell. AB - This communication reports the design and characterization of an air-breathing laminar flow-based microfluidic fuel cell (LFFC). The performance of previous LFFC designs was cathode-limited due to the poor solubility and slow transport of oxygen in aqueous media. Introduction of an air-breathing gas diffusion electrode as the cathode addresses these mass transfer issues. With this design change, the cathode is exposed to a higher oxygen concentration, and more importantly, the rate of oxygen replenishment in the depletion boundary layer on the cathode is greatly enhanced as a result of the 4 orders of magnitude higher diffusion coefficient of oxygen in air as opposed to that in aqueous media. The power densities of the present air-breathing LFFCs are 5 times higher (26 mW/cm2) than those for LFFCs operated using formic acid solutions as the fuel stream and an oxygen-saturated aqueous stream at the cathode ( approximately 5 mW/cm2). With the performance-limiting issues at the cathode mitigated, these air-breathing LFFCs can now be further developed to fully exploit their advantages of direct control over fuel crossover and the ability to individually tailor the chemical composition of the cathode and anode media to enhance electrode performance and fuel utilization, thus increasing the potential of laminar flow-based fuel cells. PMID- 16316202 TI - A rhodamine-based fluorescent and colorimetric chemodosimeter for the rapid detection of Hg2+ ions in aqueous media. AB - A rhodamine-based fluorescent and colorimetric chemodosimeter for the rapid detection of Hg2+ ions in aqueous media was developed. The system, which utilizes an irreversible Hg2+-promoted oxadiazole forming reaction, responds instantaneously at room temperature in a 1:1 stoichiometric manner to the amount of Hg2+. The selectivity of this system for Hg2+ over other metal ions is remarkably high, and its sensitivity is below 2 ppb in aqueous solutions. PMID- 16316203 TI - Bifunctional activation and racemization in the catalytic asymmetric Aza-Baylis Hillman reaction. AB - The mechanism of bifunctional activation in the asymmetric aza-Baylis-Hillman (aza-BH) reaction was studied using NMR spectroscopic techniques. The reaction involves rate-limiting proton transfer in the absence of added protic species, but exhibits no autocatalysis. Bronsted acidic additives lead to substantial rate enhancements through acceleration of the elimination step. Furthermore, it was found that phosphine catalysts either alone or in combination with protic additives can cause racemization of the aza-BH product by proton exchange at the stereogenic center. This indicates that the spatial arrangement of a bifunctional chiral catalyst for the asymmetric aza-BH reaction is crucial not only for the stereodifferentiation within the catalytic cycle but also for the prevention of subsequent racemization. PMID- 16316204 TI - Tetraplatinated artificial oligopeptides afford high affinity intercalation into dsDNA. AB - This paper reports the binding of an artificial tetrapeptide to which are tethered four Pt(II) complexes (i.e., [Pt(tpy)(py)]48+) with a 12 base pair duplex DNA oligonucleotide. Isothermal Titration Calorimetry reveals that two tetrametallic peptides stoichiometrically bind to each DNA duplex with a binding constant, KB, of 1.7 x 106 M-1, with a change in free energy of -8.5 kcal/mol. This KB represents an affinity 2 orders of magnitude greater than that of the monometallic analogue [Pt(tpy)(pic)]2+ for the same dsDNA sequence. The metalated peptides bind by intercalation into the DNA, partially unwinding the helix while stabilizing the structure, causing an increase in the dsDNA melting temperature of 25 degrees C. PMID- 16316205 TI - Equilibrium/nonequilibrium initial configurations in forward/reverse electron transfer within mixed-metal hemoglobin hybrids. AB - In a protein-protein electron transfer (ET) photocycle, the "forward" ET reaction is initiated with the excited complex, [3DA], in an equilibrium ensemble of configurations, the majority of which exhibit less than the maximal ET matrix element. In contrast, the charge-separated intermediate complex is formed in a nonequilibrium set of configurations with maximal ET matrix elements and would be expected to return to the ground state with the largest rate constant possible unless conformational interconversion first "breaks the connection" and the complex converts to less-reactive substates. According to this analysis, the forward and back ET reactions should show a differential response to viscosity, and the latter could even show an increased rate constant under conditions which suppress departure from the reactive configuration(s). We now report that the viscosity dependences of forward and back ET rate constants for the photocycle within the [alpha2(Zn),beta2(Fe3+N3-)] mixed-metal hemoglobin hybrid at pH 7 show the anticipated behavior: kf decreases as viscosity increases, but, in sharp contrast, kb increases strongly. PMID- 16316206 TI - The bis-urea motif as a tool to functionalize self-assembled nanoribbons. AB - Here we present a surfactant molecule (1) containing an ammonium headgroup, in which a bis-ureido group is incorporated in its hydrocarbon chain. Due to strong hydrogen bonding interactions, 1 forms well-defined highly ordered ribbon-like aggregates in water. Moreover, we demonstrate that these ribbons can be functionalized via a modular approach through molecular recognition of other bis urea containing molecules. The dye disperse orange and biotin were coupled to matching bis-ureido groups and incorporated into the ribbon structure. The anchoring of different functionalities in a modular approach proved to be possible using the molecular recognition capabilities of the bis-ureido moiety, thereby opening possibilities to a wide range of applications. PMID- 16316207 TI - Absolute molecular weight of polyaniline. AB - The absolute molecular weight of polyaniline in the pernigraniline, emeraldine, and leucoemeraldine oxidation states has been measured by light scattering and the exact number of aniline repeat units determined for the first time. Using potential-time profiling to monitor the chemical oxidative polymerization of aniline using ammonium peroxydisulfate oxidant, all three oxidation states of polyaniline can be synthesized in one step and the evolution of polymer molecular weight monitored. The pernigraniline intermediate formed during the chemical oxidative polymerization of aniline increases by 17-20% when it is converted to emeraldine, which is consistent with a two-step polymerization mechanism. These findings establish a solid experimental framework to chemically synthesize block copolymers of polyaniline by using different monomers to intercept the reaction at the pernigraniline oxidation state. PMID- 16316208 TI - Halobenzenes and Ir(I): kinetic C-H oxidative addition and thermodynamic C-Hal oxidative addition. AB - A (PNP)Ir fragment undergoes facile, room-temperature oxidative addition of C-H bonds in arenes and haloarenes in preference to aromatic carbon-halogen bonds. This preference, however, is determined to be kinetic in nature. Oxidative addition of C-Cl and C-Br is preferred thermodynamically. The products of the C Cl or C-Br oxidative addition are separated from the C-H oxidative addition products by a high activation barrier and are only accessible at >100 degrees C. Of the C-H oxidative addition products of chlorobenzene, the isomer with the o ClC6H4 ligand has the lowest energy. PMID- 16316209 TI - High resolution printing of DNA feature on poly(methyl methacrylate) substrates using supramolecular nano-stamping. AB - In recent years, a large number of devices based on organic and biological materials have been developed. To scale-up the production of these systems to industrially acceptable standards, there is a need to develop soft-material stamping approaches with the needed resolution, complexity, and versatility. We have recently developed a DNA-based stamping method (supramolecular nano stamping, SuNS) that has superior resolution and can print multiple molecules at the same time. A similar technique was independently developed by Crooks and co workers. Here we show that SuNS can be used to efficiently print DNA features on a polymeric substrate (poly(methyl methacrylate), PMMA) with a 40 nm point resolution and a coverage that exceeds 100 mum2. The stamped PMMA substrate was also used as a master to print on a gold substrate. With PMMA being optically clear and electrically insulating, future applications of SuNS to print DNA micro and nanoarrays are envisioned. PMID- 16316210 TI - A cobalt(III)-salen complex with an axial substituent in the diamine backbone: stereoselective recognition of amino alcohols. AB - A cobalt(III)-salen complex (3) with an axial substituent on the diamine backbone has been synthesized. Crystal structure reveals that the axial substituent (p nitrophenyl group) is positioned in close proximity to the metal binding site. The stereoselectivity of the cobalt complex for binding amino alcohols increases with increasing steric bulk of the amino alcohol from alaninol (2.9) to valinol (6.2) and t-leucinol (36.0). PMID- 16316211 TI - Remarkable phosphine-effect on the intramolecular aldol reactions of unsaturated 1,5-diketones: highly regioselective synthesis of cross-conjugated dienones. AB - We report a phosphine-mediated intramolecular aldol cyclization of unsaturated diketones that proceeds with extremely high levels of regioselectivity for the cross-conjugated bicyclic dienone products. The sense of regioselectivity observed in this reaction is complementary to that obtained using traditional aldol conditions. Experimental evidence that supports the involvement of a phosphine Michael adduct is described. PMID- 16316212 TI - N2 provides insight into the mechanism of H-C(sp3) bond cleavage. AB - Exchange of deuterium in d6-benzene with all C-H sites in (PNP)Ru(OTf), where PNP is N(SiMe2CH2PtBu2)2 and OTf is OSO2CF3, is rapid at 22 degrees C. Although intact planar triplet (PNP)Ru(OTf) binds N2 only very weakly, these reagents are observed to react rapidly to give a diamagnetic 1:1 adduct whose structure has one tBu C-H bond cleaved: the carbon binds to Ru but the hydrogen is on the PNP nitrogen, creating a secondary amine ligand bound to RuII. It is suggested that the benzene C-D cleavage and the N2 product of tBu C-H bond heterolysis both derive from a common intermediate, [HN(SiMe2CH2PtBu2)(SiMe2CH2PtBuCMe2CH2)] Ru(OTf); the formation energy and structure of this species are discussed on the basis of DFT results. PMID- 16316213 TI - Synthesis of alpha-P-modified nucleoside diphosphates with ethylenediamine. AB - This report describes a one-pot synthesis of alpha-P-borano-, alpha-P-thio-, and alpha-P-seleno-modified nucleoside diphosphate analogues that are otherwise difficult to obtain. The key step involves the intramolecular nucleophilic attack by an amino group in 5 to remove the gamma-phosphate. The absolute configurations of P-diastereomers were confirmed by analysis of their 1H NMR. Affinity studies revealed that the nucleoside boranodiphosphates are potentially useful in antiviral research. PMID- 16316214 TI - Preparation and characterization of unique inorganic-organic hybrid mesoporous materials incorporating arenetricarbonyl complexes [-C6H4M(CO)3-] (M = Cr, Mo). AB - Unique inorganic-organic hybrid mesoporous materials incorporating arenetricarbonyl complexes [-C6H4M(CO)3-] (M = Cr, Mo) within their mesoporous frameworks have been successfully prepared by a simple CVD treatment of the phenylene-bridged organosilica mesoporous materials with metal hexacarbonyls [M(CO)6] (M = Cr, Mo). FT-IR and UV-vis investigations revealed that the arenetricarbonyl complexes exist stably even at temperatures above 473 K under vacuum. PMID- 16316215 TI - Controlling molecular diffusion in self-spreading lipid bilayer using periodic array of ultra-small metallic architecture on solid surface. AB - Diffusion of target molecules incorporated in the self-spreading lipid bilayer was controlled by the introduction of periodic array of metallic architecture on solid surface. Retardation of the progress of target molecules became significant when the size of gap between small metal architectures was less than a few hundred nanometers. The self-spreading dynamics of the lipid bilayer depending on the size of the small gap were analyzed semiquantitatively. Estimated change in the driving force of the spreading layer suggests that highly localized compression of the spreading layer causes selective segregation of molecules. PMID- 16316216 TI - Catalytic addition of terminal alkynes to carbodiimides by half-sandwich rare earth metal complexes. AB - The catalytic addition of terminal alkynes to carbodiimides has been achieved for the first time by use of half-sandwich rare earth metal complexes, such as {Me2Si(C5Me4)(NPh)}Y(CH2SiMe3)(THF)2, which offers a straightforward, atom economic route to the N,N'-disubstituted propiolamidines which contain a conjugated C-C triple bond, a new family of amidines which were difficult to prepare by other means. A rare earth metal amidinate species was confirmed to be a true catalytic species in this process, thus demonstrating for the first time that an amidinate unit, though being often used as an ancillary ligand for various organometallic complexes, can itself participate in a catalytic reaction under appropriate conditions. PMID- 16316217 TI - Zirconium-mediated SN2' substitution of allylic ethers: regio- and stereospecific formation of protected allylic amines. AB - A new zirconium-mediated, regio- and stereospecific SN2' substitution of allylic ethers with a nitrogen nucleophile has been developed. Cbz-protected amine products were isolated in high yield from reactions with a wide range of Z allylic ethers. A mechanism of the allylic substitution consistent with the results of the kinetics and kinetic isotope effect studies was proposed. PMID- 16316218 TI - In situ crystallization of low-melting ionic liquids. AB - Single crystals of five very low-melting ionic liquids, [emim]BF4 (mp -1.3 degrees C), [bmim]PF6 (+1.9 degrees C), [bmim]OTf (+6.7 degrees C), [hexpy]NTf2 ( 3.6 degrees C), and [bmpyr]NTf2 (-10.8 degrees C), have been grown using a combined calorimetric and zone-melting approach and their crystal structures determined by X-ray diffraction. PMID- 16316219 TI - Homopolymer micelles in heterogeneous solvent mixtures. AB - Amphiphilic homopolymers containing a hydrophilic and a hydrophobic functionality in each monomer unit have been shown to form polar or apolar containers depending on the solvent environment. When presented with a mixture of solvents, these polymeric containers are capable of releasing certain guest molecules. The fundamental mechanism behind these properties is investigated, and the utility of these assemblies in separations has been demonstrated with an example. PMID- 16316220 TI - Synthesis of (-)-quinocarcin by directed condensation of alpha-amino aldehydes. AB - An enantioselective synthesis of the natural antiproliferative agent quinocarcin was achieved by the directed condensation of optically active alpha-amino aldehyde intermediates. Condensation of the N-protected alpha-amino aldehyde 1, prepared in eight steps (19% yield) from (R,R)-pseudoephedrine glycinamide, with the C-protected alpha-amino aldehyde derivative 2, prepared in seven steps (34% yield) from (R,R)-pseudoephedrine glycinamide, afforded the corresponding imine in quantitative yield. Without isolation, direct treatment of this imine intermediate with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) and hydrogen cyanide led to cleavage of the fluorenylmethoxycarbonyl (Fmoc) protective group followed by addition of cyanide (Strecker reaction) to form the bis-amino nitriles 3 as a mixture of diastereomers, in 91% yield. Treatment of the diastereomers 3 with trimethylsilyl cyanide and zinc chloride in 2,2,2-trifluoroethanol at 60 degrees C led to stepwise cyclization to form the tetracyclic product 4 (42% yield from 1 and 2). The latter intermediate was transformed into (-)-quinocarcin (1) in five steps (45% yield). The yield of quinocarcin was 19% from 1 and 2 (7 steps), and 4% from pseudoephedrine glycinamide (15 steps). PMID- 16316221 TI - Cucurbit[10]uril. AB - Melamine diamine 1 is able to displace CB[5] from the CB[10].CB[5] complex resulting in CB[10].12 and precipitated CB[5].1. We were able to isolate free CB[10] by treatment of CB[10].1 with acetic anhydride followed by washing with MeOH, DMSO, and water. The spacious cavity of CB[10] is able to complex large guests, including a cationic calix[4]arene derivative in its 1,3-alternate form (CB[10].1,3-alt-3). The addition of adamantane carboxylic acid (4) to CB[10].3 triggers a conformational change during the formation of termolecular complex CB[10].cone-3.4. PMID- 16316222 TI - A selective, noncovalent assay for base flipping in DNA. AB - Base flipping, the conformational change of a nucleobase to an extrahelical position, is a key step in the enzymatic repair of damaged DNA. An assay that can detect the flipped-out species in free solution without covalent modification of the DNA would be desirable. The design and synthesis of a simple, sensitive, and rapid assay using specific noncovalent binding to pyrimidines by zinc-cyclen and a commonly used fluorescent reporter group, dansyl, is reported. The binding of the zinc-cyclen unit to a flipped-out thymine base results in a change in the fluorescent properties of the dansyl group that is distinct from nonspecific binding to duplex DNA or intercalation into either the flipped-in or flipped-out species. The assay was tested using fluorescence spectroscopy and detection at 533 +/- 5 nm with normal and abasic duplex DNA as negative and positive controls. The data obtained are fitted to a one-site binding model to determine the equilibrium constant for the two-step process involving base flipping and binding to be approximately 10-6 M. PMID- 16316223 TI - The predictably elusive form II of aspirin. AB - The elusive form II of aspirin has been obtained during co-crystallization experiments with levetiracetam or acetamide, and it has been characterized by IR, DSC, HPLC, and single-crystal X-ray diffraction. PMID- 16316224 TI - Tandem au-catalyzed 3,3-rearrangement-[2 + 2] cycloadditions of propargylic esters: expeditious access to highly functionalized 2,3-indoline-fused cyclobutanes. AB - The treatment of readily available propargylic indole-3-acetates with a catalytic amount of AuCl(PPh3)/AgSbF6 leads to tandem activations of the propargylic esters and the in situ generated allenylic esters, resulting in expeditious access to highly functionalized cyclobutanes with fused 2,3-indoline and gamma-lactone rings and an exocyclic E-double bond through sequential 3,3-rearrangement and [2 + 2] cyclization. PMID- 16316225 TI - SO2:H2O surface complex found at the vapor/water interface. AB - A weakly bonded SO2:H2O surface complex is found at the vapor/water interface prior to the reaction and dissolution of SO2 into the aqueous phase. The results have important implications for understanding the formation of atmospheric aerosols and understanding the atmospheric sulfur cycle. PMID- 16316226 TI - Direct imaging and spectroscopic characterization of stimulus-responsive microgels. AB - Scanning transmission X-ray microscopy has been employed to visualize pH responsive acid-swellable microgel particles directly in their swollen state in aqueous acidic solution. Moreover, NEXAFS studies confirm that the nitrogen atoms of these cationic microgel particles are completely protonated at low pH. PMID- 16316227 TI - Determination of the size distribution of sonoluminescence bubbles in a pulsed acoustic field. AB - A simple method is described for determining the size of sonoluminescence bubbles generated by acoustic cavitation. The change in the intensity of sonoluminescence, from 4 ms pulses of 515 kHz ultrasound, as a function of the "off" time between acoustic pulses, is the basis of the method. The bubble size determined in water was in the range of 2.8-3.7 mum. PMID- 16316229 TI - The fate of dicationic states in molecular clusters of benzene and related compounds. AB - Calculations employing density functional theory indicate that, rather than undergoing fragmentation, dicationic clusters of benzene, hexafluorobenzene, and naphthalene produced by sequential one-electron or sudden double-ionization experiments on the neutrals can relax via the formation of inter-ring covalent C C bonds, along with a series of proton transfers that enable a substantial reduction of inter- and intramolecular Coulomb repulsions. The theoretically predicted chemically bound structures correspond to deep local energy minima on the potential energy surface pertaining to the lowest electronic state of the dications and can therefore be regarded as metastable (kinetically long-lived) species. This discovery invalidates on theoretical grounds the liquid-droplet model of multiply charged clusters and sheds very unexpected light on possible consequences in chemistry of the intermolecular Coulombic decay (ICD) mechanism [Cederbaum, L. S.; et al. Phys. Rev. Lett. 1997, 79, 4778; Jahnke, T.; et al. Phys. Rev. Lett. 2004, 93, 163401] for deep inner-valence ionized states. Propagation of charge rearrangement reactions and proton transfers to several monomers may eventually lead to the formation of rather extended dicationic assemblies. PMID- 16316228 TI - QZ1 and QZ2: rapid, reversible quinoline-derivatized fluoresceins for sensing biological Zn(II). AB - QZ1, 2-[2-chloro-6-hydroxy-3-oxo-5-(quinolin-8-ylaminomethyl)-3H-xanthen-9 yl]benzoic acid, and QZ2, 2-[6-hydroxy-3-oxo-4,5-bis-(quinolin-8-ylaminomethyl) 3H-xanthen-9-yl]benzoic acid, two fluorescein-based dyes derivatized with 8 aminoquinoline, have been prepared and their photophysical, thermodynamic, and zinc-binding kinetic properties determined. Because of their low background fluorescence and highly emissive Zn(II) complexes, QZ1 and QZ2 have a large dynamic range, with approximately 42- and approximately 150-fold fluorescence enhancements upon Zn(II) coordination, respectively. These dyes have micromolar K(d) values for Zn(II) and are selective for Zn(II) over biologically relevant concentrations of the alkali and alkaline earth metals. The Zn(II) complexes also fluoresce brightly in the presence of excess Mn(II), Fe(II), Co(II), Cd(II), and Hg(II), offering improved specificity for Zn(II) over di(2-picolyl)amine-based Zn(II) sensors. Stopped-flow kinetic investigations indicate that QZ1 and QZ2 bind Zn(II) with k(on) values of (3-4) x 10(6) M(-1) s(-1), compared to (6-8) x 10(5) M(-1) s(-1) for select ZP (Zinpyr) dyes, at 4.3 degrees C. Dissociation of Zn(II) from QZ1 and QZ2 occurs with k(off) values of 150 and 160 s(-1), over 5 orders of magnitude larger than those for ZP probes, achieving reversibility on the biological (millisecond) time scale. Laser scanning confocal and two-photon microscopy studies reveal that QZ2 is cell-permeable and Zn(II)-responsive in vivo. Because of its weaker affinity for Zn(II), QZ2 responds to higher concentrations of intracellular Zn(II) than members of the ZP family, illustrating that binding affinity is an important parameter for Zn(II) detection in vivo. PMID- 16316230 TI - Combined experimental and DFT-TDDFT computational study of photoelectrochemical cell ruthenium sensitizers. AB - We report a combined experimental and computational study of several ruthenium(II) sensitizers originated from the [Ru(dcbpyH(2))(2)(NCS)(2)], N3, and [Ru(dcbpyH(2))(tdbpy)(NCS)(2)], N621, (dcbpyH(2) = 4,4'-dicarboxy-2,2' bipyridine, tdbpy = 4,4'-tridecyl-2,2'-bipyridine) complexes. A purification procedure was developed to obtain pure N-bonded isomers of both types of sensitizers. The photovoltaic data of the purified N3 and N621 sensitizers adsorbed on TiO(2) films in their monoprotonated and diprotonated state, exhibited remarkable power conversion efficiency at 1 sun, 11.18 and 9.57%, respectively. An extensive Density Functional Theory (DFT)-Time Dependent DFT study of these sensitizers in solution was performed, investigating the effect of protonation of the terminal carboxylic groups and of the counterions on the electronic structure and optical properties of the dyes. The calculated absorption spectra are in good agreement with the experiment, thus allowing a detailed assignment of the UV-vis spectral features of the two types of dyes. The computed alignments of the molecular orbitals of the different complexes with the band edges of a model TiO(2) nanoparticle provide additional insights into the electronic factors governing the efficiency of dye-sensitized solar cell devices. PMID- 16316231 TI - Hydration state of nonionic surfactant monolayers at the liquid/vapor interface: structure determination by vibrational sum frequency spectroscopy. AB - The OH stretching region of water molecules in the vicinity of nonionic surfactant monolayers has been investigated using vibrational sum frequency spectroscopy (VSFS) under the polarization combinations ssp, ppp, and sps. The surface sensitivity of the VSFS technique has allowed targeting the few water molecules present at the surface with a net orientation and, in particular, the hydration shell around alcohol, sugar, and poly(ethylene oxide) headgroups. Dramatic differences in the hydration shell of the uncharged headgroups were observed, both in comparison to each another and in comparison to the pure water surface. The water molecules around the rigid glucoside and maltoside sugar rings were found to form strong hydrogen bonds, similar to those observed in tetrahedrally coordinated water in ice. In the case of the poly(ethylene oxide) surfactant monolayer a significant ordering of both strongly and weakly hydrogen bonded water was observed. Moreover, a band common to all the surfactants studied, clearly detected at relatively high frequencies in the polarization combinations ppp and sps, was assigned to water species located in proximity to the surfactant hydrocarbon tail phase, with both hydrogen atoms free from hydrogen bonds. An orientational analysis provided additional information on the water species responsible for this band. PMID- 16316232 TI - Tubular configurations and structure-dependent anisotropic strains in GaS multi walled sub-microtubes. AB - Bending strains and bonding structures of GaS multiwalled submicrotubes have been examined by transmission electron microscopy. Experimental observations reveal that the strain involved in building the GaS tubes is more complicated than the theoretical prediction and appears anisotropic and dependent on the tubular configurations. The armchair tube bears a larger lattice compression than the zigzag tube. The hexagonal GaS semiconducting compound degrades its crystal symmetry upon bending, leading to the anisotropy of the bonding structures. Curving of GaS sheets to form tubes is found to be dominated by the structures and electrostatic fields on the sheet surface, which eventually gives a well controlled tubular structure showing preferred zigzag and close-to-zigzag configurations. Finally, interlayer packing structures of multiwalled tubes are examined. PMID- 16316233 TI - Intermolecular charge transfer between heterocyclic oligomers. Effects of heteroatom and molecular packing on hopping transport in organic semiconductors. AB - For electron or hole transfer between neighboring conducting polymer strands or oligomers, the intrinsic charge-transfer rate is dictated by the charge-resonance integral and by the reorganization energy due to geometric relaxation. To explain conduction anisotropy and other solid-state effects, a multivariate, systematic analysis of bandwidth as a function of intermolecular orientations is undertaken for a series of oligoheterocycles, using first-principles methods. While cofacial oligomers show the greatest bandwidths at a given intermolecular C-C contact distance, for a fixed center-to-center intermolecular distance, tilted pi stacking increases pi-overlap (particularly for LUMO orbitals) and decreases electrostatic repulsion, yielding optimum tilt angles for packing of approximately 40-60 degrees at small intermolecular separations. The calculations also reveal that bandwidths and intrinsic mobilities of holes and electrons in conjugated oligoheterocycles can be quite comparable. PMID- 16316234 TI - Spectroscopic studies of the anaerobic enzyme-substrate complex of catechol 1,2 dioxygenase. AB - The basis of the respective regiospecificities of intradiol and extradiol dioxygenase is poorly understood and may be linked to the protonation state of the bidentate-bound catechol in the enzyme/substrate complex. Previous ultraviolet resonance Raman (UVRR) and UV-visible (UV-vis) difference spectroscopic studies demonstrated that, in extradiol dioxygenases, the catechol is bound to the Fe(II) as a monoanion. In this study, we use the same approaches to demonstrate that, in catechol 1,2-dioxygenase (C12O), an intradiol enzyme, the catechol binds to the Fe(III) as a dianion. Specifically, features at 290 nm and 1550 cm(-1) in the UV-vis and UVRR difference spectra, respectively, are assigned to dianionic catechol based on spectra of the model compound, ferric tris(catecholate). The UVRR spectroscopic band assignments are corroborated by density functional theory (DFT) calculations. In addition, negative features at 240 nm in UV-vis difference spectra and at 1600, 1210, and 1175 cm(-1) in UVRR difference spectra match those of a tyrosinate model compound, consistent with protonation of the axial tyrosinate ligand when it is displaced from the ferric ion coordination sphere upon substrate binding. The DFT calculations ascribe the asymmetry of the bound dianionic substrate to the trans donor effect of an equatorially ligated tyrosinate ligand. In addition, the computations suggest that trans donation from the tyrosinate ligand may facilitate charge transfer from the substrate to yield the iron-bound semiquinone transition state, which is capable of reacting with dioxygen. In illustrating the importance of ligand trans effects in a biological system, the current study demonstrates the power of combining difference UVRR and optical spectroscopies to probe metal ligation in solution. PMID- 16316235 TI - Shell click-crosslinked (SCC) nanoparticles: a new methodology for synthesis and orthogonal functionalization. AB - A new methodology for the preparation of well-defined core-shell nanoparticles was developed, based upon the employment of a multifunctional crosslinker to coincidently stabilize supramolecular polymer assemblies and imbed into the shell unique chemical functionalities. Amphiphilic diblock copolymers of poly(acrylic acid)(80)-b-poly(styrene)(90) that had been assembled into micelles and partially functionalized throughout the corona with alkynyl groups were utilized as Click readied nanoscaffolds for the formation of shell Click-crosslinked nanoparticles (SCCs). Divergently grown dendrimers of the zero, first, second, and third generations having increasing numbers of azide terminating groups ((N(3))(2)-[G 0], (N(3))(4)-[G-1], (N(3))(8)-[G-2], and (N(3))(16)-[G-3], respectively) were investigated as crosslinkers via Click reactions with the alkynyl groups to form covalent linkages throughout the block copolymer micelle corona, thus forming a crosslinked shell. The crosslinking reactions were characterized by (1)H NMR and IR spectroscopies, differential scanning calorimetry (DSC), and dynamic light scattering (DLS) measurements. Only the first generation dendrimer ((N(3))(4)-[G 1]) possessed a sufficient balance of polyvalency and water solubility to achieve crosslinking and establish a robust nanostructure. The resulting SCC was further characterized with atomic force microscopy (AFM), transmission electron microscopy (TEM), and analytical ultracentrifugation (AU). The dendritic crosslinker is important as it also allows for the incorporation of excess functionality that can undergo complementary reactions. Within the shell of this SCC the remaining azide termini of the dendrimer crosslinker were then consumed in a secondary Click reaction with an alkynyl-functionalized fluorescein to yield a fluorescently labeled SCC that was characterized with DLS, AFM, TEM, AU, UV vis, and fluorescent measurements as a function of pH. PMID- 16316236 TI - Intervalence transitions in the mixed-valence monocations of bis(triarylamines) linked with vinylene and phenylene-vinylene bridges. AB - (E)-4,4'-Bis[bis(4-methoxyphenyl)amino]stilbene, 1, (E,E)-1,4-bis[4-[bis(4 methoxyphenyl)amino]styryl]benzene, 2, and two longer homologues, (E,E,E)-4,4' bis[4-[bis(4-methoxyphenyl)amino]styryl]stilbene, 3, and (E,E,E,E)-1,4-bis(4-[4 [bis(4-methoxyphenyl)amino]styryl]styryl)benzene, 4, have been oxidized to their mono- and dications using tris(4-bromophenyl)aminium hexachloroantimonate. The intervalence charge-transfer (IVCT) band of 1(+) is narrow and asymmetric and exhibits only weak solvatochromism. Analysis of this band indicates that 1(+) is a class-III or class-II/III borderline mixed-valence species. In contrast, a broad, strongly solvatochromic IVCT band is observed for 2(+), indicating that this species is a class-II mixed-valence species. The assignment of 1(+ ) and 2(+) as symmetric class-III and unsymmetric class-II species, respectively, is also supported by AM1 calculations. Hush analysis of the IVCT bands of both 1(+) and 2(+) gives larger electronic couplings, V, than for their analogues in which the double bonds are replaced with triple bonds. The diabatic electron-transfer distance, R, in 1(+) can be estimated by comparison of the V estimated by Hush analysis and from the IVCT maximum; it is considerably less than the geometric N N separation, a result supported by quantum-chemical estimates of R for 1(+) 4(+). In 3(+) and 4(+), the IVCT is largely obscured by an intense absorption similar to a band seen in the corresponding dications and to that observed in the monocation of a model compound, (E,E,E)-1-[bis(4-methoxyphenyl)amino]-4-[4-[4-(4 tert-butylstyryl)styryl]styryl]benzene, 5, containing only one nitrogen redox center; we attribute this band to a bridge-to-N(+) transition. The corresponding dications 1(2+)-4(2+) show a complementary trend in the coupling between redox centers: the shortest species is diamagnetic, while the dication with the longest bridge behaves as two essentially noninteracting radical centers. PMID- 16316237 TI - N-triazinylammonium tetrafluoroborates. A new generation of efficient coupling reagents useful for peptide synthesis. AB - A new generation of triazine-based coupling reagents (TBCRs), designed according to the concept of "superactive esters", was obtained by treatment of 4-(4,6 dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium (DMTMM) chloride with lithium or silver tetrafluoroborate. The structure of 4-(4,6-dimethoxy-1,3,5-triazin-2 yl)-4-methylmorpholinium tetrafluoroborate was confirmed by X-ray diffraction. Activation of carboxylic acids by using this reagent proceeds via triazine "superactive ester". The coupling reagent was successfully used for the synthesis of Z-, Boc-, and Fmoc-protected dipeptides derived from natural and unnatural sterically hindered amino acids and for fragment condensation, in 80-100% yield and with high enantiomeric purity. The manual SPPS of the ACP(65-74) peptide fragment (H-Val-Gln-Ala-Ala-Ile-Asp-Tyr-Ile-Asn-Gly-OH) proceeded significantly faster than with TBTU or HATU, as well as the automated SPPS of the same fragment gave a purer product than by using TBTU or PyBOP. The reagent was also demonstrated to be efficient in on-resin head-to-tail cyclization of constrained cyclopeptides, in SPPS synthesis of Aib peptides, and in the synthesis of esters from appropriate acids, alcohols, and phenols. The high efficiency and versatility of this new generation of TBCRs confirm, for the first time, the usefulness of the concept of "superactive esters" in rational design of the structure of coupling reagents. PMID- 16316239 TI - Nature of the chemical bond between metal atoms and oxide surfaces: new evidences from spin density studies of K atoms on alkaline earth oxides. AB - We have studied the interaction of K atoms with the surface of polycrystalline alkaline-earth metal oxides (MgO, CaO, SrO) by means of CW- and Pulsed-EPR, UV Vis-NIR spectroscopies and DFT cluster model calculations. The K adsorption site is proposed to be an anionic reverse corner formed at the intersection of two steps, where K binds by more than 1 eV, resulting in thermally stable species up to about 400 K. The bonding has small covalent and large polarization contributions, and the K atom remains neutral, with one unpaired electron in the valence shell. The interaction results in strong modifications of the K electronic wave function which are directly reflected by the hyperfine coupling constant, (K)a(iso). This is found to be a very efficient "probe" to measure the degree of metal-oxide interaction which directly depends on the substrate basicity. These results provide an original and general model of the early stages of the metal-support interaction in the case of ionic oxides. PMID- 16316238 TI - Temperature-dependent studies of NO recombination to heme and heme proteins. AB - The rebinding kinetics of NO to the heme iron of myoglobin (Mb) is investigated as a function of temperature. Below 200 K, the transition-state enthalpy barrier associated with the fastest (approximately 10 ps) recombination phase is found to be zero and a slower geminate phase (approximately 200 ps) reveals a small enthalpic barrier (approximately 3 +/- 1 kJ/mol). Both of the kinetic rates slow slightly in the myoglobin (Mb) samples above 200 K, suggesting that a small amount of protein relaxation takes place above the solvent glass transition. When the temperature dependence of the NO recombination in Mb is studied under conditions where the distal pocket is mutated (e.g., V68W), the rebinding kinetics lack the slow phase. This is consistent with a mechanism where the slower (approximately 200 ps) kinetic phase involves transitions of the NO ligand into the distal heme pocket from a more distant site (e.g., in or near the Xe4 cavity). Comparison of the temperature-dependent NO rebinding kinetics of native Mb with that of the bare heme (PPIX) in glycerol reveals that the fast (enthalpically barrierless) NO rebinding process observed below 200 K is independent of the presence or absence of the proximal histidine ligand. In contrast, the slowing of the kinetic rates above 200 K in MbNO disappears in the absence of the protein. Generally, the data indicate that, in contrast to CO, the NO ligand binds to the heme iron through a "harpoon" mechanism where the heme iron out-of-plane conformation presents a negligible enthalpic barrier to NO rebinding. These observations strongly support a previous analysis (Srajer et al. J. Am. Chem. Soc. 1988, 110, 6656-6670) that primarily attributes the low temperature stretched exponential rebinding of MbCO to a quenched distribution of heme geometries. A simple model, consistent with this prior analysis, is presented that explains a variety of MbNO rebinding experiments, including the dependence of the kinetic amplitudes on the pump photon energy. PMID- 16316240 TI - Effects of turn stability on the kinetics of refolding of a hairpin in a beta sheet. AB - As part of our continuing study of the effects of the turn sequence on the conformational stability as well as the mechanism of folding of a beta-sheet structure, we have undertaken a parallel investigation of the solution structure, conformational stability, and kinetics of refolding of the beta-sheet VFIVDGOTYTEV(D)PGOKILQ. The latter peptide is an analogue of the original Gellman beta-sheet VFITS(D)PGKTYTEV(D)PGOKILQ, wherein the TS(D)PGK turn sequence in the first hairpin has been replaced by VDGO. Thermodynamics studies revealed comparable conformational stability of the two peptides. However, unlike the Gellman peptide, which showed extremely rapid refolding of the first hairpin, early kinetic events associated with the refolding of the corresponding hairpin in the VDGO mutant were found to be significantly slower. A detailed study of the conformation of the modified peptide suggested that hydrophobic interactions might be contributing to its stability. Accordingly, we surmise that the early kinetic events are sensitive to whether the formation of the hairpin is nucleated at the turn or by sequestering of the hydrophobic residues across the strand, before structural rearrangements to produce the nativelike topology. Nucleation of the hairpin at the turn is expected to be intrinsically rapid for a strong turn. However, if the process must involve collapse of hydrophobic side chains, the nucleation should be slower as solvent molecules must be displaced to sequester the hydrophobic residues. These findings reflect the contribution of different forces toward nucleation of hairpins in the mechanism of folding of beta-sheets. PMID- 16316241 TI - In situ preparation of protein-"smart" polymer conjugates with retention of bioactivity. AB - Protein-polymer conjugates are widely used in biotechnology and medicine, and new methods to prepare the bioconjugates would be advantageous for these applications. In this report, we demonstrate that bioactive "smart" polymer conjugates can be synthesized by polymerizing from defined initiation sites on proteins, thus preparing the polymer conjugates in situ. In particular, free cysteines, Cys-34 of bovine serum albumin (BSA) and Cys-131 of T4 lysozyme V131C, were modified with initiators for atom transfer radical polymerization (ATRP) either through a reversible disulfide linkage or irreversible bond by reaction with pyridyl disulfide- and maleimide-functionalized initiators, respectively. Initiator conjugation was verified by electrospray-ionization mass spectroscopy (ESI-MS), and the location of the modification was confirmed by muLC-MSMS (tandem mass spectrometry) analysis of the trypsin-digested protein macroinitiators. Polymerization of N-isopropylacrylamide (NIPAAm) from the protein macroinitiators resulted in thermosensitive BSA-polyNIPAAm and lysozyme-polyNIPAAm in greater than 65% yield. The resultant conjugates were characterized by gel electrophoresis and size exclusion chromatography (SEC) and easily purified by preparative SEC. The identity of polymer isolated from the BSA conjugate was confirmed by (1)H NMR, and the polydispersity index was determined by gel permeation chromatography (GPC) to be as low as 1.34. Lytic activities of the lysozyme conjugates were determined by two standard assays and compared to that of the unmodified enzyme prior to polymerization; no statistical differences in bioactivity were observed. PMID- 16316242 TI - Energetic characterization of short helical polyalanine peptides in water: analysis of 13C=O chemical shift data. AB - Measured at 2 degrees C in water, NMR chemical shifts of (13)C=O labeled central alanine residues of peptides W-Lys(5)-(t)L(3)-Ala(n)-(t)L(3)-Lys(5)NH(2), n = 9, 11, 13, 15, 19 and W-Lys(5)-(t)L(3)-a-Ala(n)-A-Inp-(t)L(2)-Lys(5)NH(2) (a = D Ala; (t)L = tert-leucine; Inp = 4-carboxypiperidine) are used to assign jt(L) and ct(L), the N- and C-terminal (t)L capping parameters and length-dependent values for w(Ala)(n), the alanine helical propensity for Ala(n) peptides. These parameters allow Lifson-Roig characterization of the stabilities of Ala(n)() helices in water. To facilitate chemical shift characterization, different (13)C/(12)C ratios are incorporated into specific Ala sites to code up to six residue sites per peptide. Large left/right chemical shift anisotropies are intrinsic to helical polyalanines, and a correcting L-R-based model is introduced. Capping parameters jt(L) = ct(L) lie in the range of 0.3 to 0.5; the (t)L residues are thus moderately helix-destabilizing. For helical conformations of lengths shorter than eight residues, assigned values for w(Ala) approach 1.0 but increase monotonically with length to a value of 1.59 for w(Ala)(19). PMID- 16316243 TI - A quantum dynamical treatment of symmetry-induced kinetic isotope effects in the formation of He2+. AB - Kinetic isotope effects for He(2)(+) formation are calculated quantum dynamically using high-quality Born-Oppenheimer (BO) potentials for two electronic states of He(2)(+) and an accurate treatment of all nonadiabatic BO corrections. The two potentials are coupled only when the helium isotopes are different, and the calculations reveal that this coupling is sufficient to allow the two sets of distinguishable reactants, (4)He(+) + (3)He or (3)He(+) + (4)He, to yield He(2)(+) with comparable efficiency over a wide temperature range. Consequently, the potential coupling provides a significant formation rate enhancement for the low isotopic symmetry reactants, as compared to the symmetrical cases (e.g., (4)He(+) + (4)He or (3)He(+) + (3)He). The computed symmetry-induced kinetic isotope effects (SIKIEs) are in substantial agreement with the available experimental results and represent the first theoretical demonstration of this unusual kinetic phenomenon. Possible application of SIKIE to ozone formation and other chemical systems is discussed. PMID- 16316244 TI - Coordinative properties of highly fluorinated solvents with amino and ether groups. AB - Despite the widespread use of perfluorinated solvents with amino and ether groups in a variety of application fields, the coordinative properties of these compounds are poorly known. It is generally assumed that the electron withdrawing perfluorinated moieties render these functional groups rather inert, but little is known quantitatively about the extent of their inertness. This paper reports on the interactions between inorganic monocations and perfluorotripentylamine and 2H-perfluoro-5,8,11-trimethyl-3,6,9,12-tetraoxapentadecane, as determined with fluorous liquid-membrane cation-selective electrodes doped with tetrakis[3,5 bis(perfluorohexyl)phenyl]borate salts. The amine does not undergo measurable association with any ion tested, and its formal pK(a) is shown to be smaller than -0.5. This is consistent with the nearly planar structure of the amine at its nitrogen center, as obtained with density functional theory calculations. The tetraether interacts very weakly with Na(+) and Li(+). Assuming 1:1 stoichiometry, formal association constants were determined to be 2.3 and 1.5 M( 1), respectively. This disproves an earlier proposition that the Lewis base character in such compounds may be nonexistent. Due to the extremely low polarity of fluorous solvents and the resulting high extent of ion pair formation, a fluorophilic electrolyte salt with perfluoroalkyl substituents on both the cation and the anion had to be developed for these experiments. In its pure form, this first fluorophilic electrolyte salt is an ionic liquid with a glass transition temperature, T(g), of -18.5 degrees C. Interestingly, the molar conductivity of solutions of this salt increases very steeply in the high concentration range, making it a particularly effective electrolyte salt. PMID- 16316245 TI - 3H-benzophosphepine complexes: versatile phosphinidene precursors. AB - The synthesis of a variety of benzophosphepine complexes [R = Ph, t-Bu, Me; ML(n )()= W(CO)(5), Mo(CO)(5), Cr(CO)(5), Mn(CO)(2)Cp] by two successive hydrophosphinations of 1,2-diethynylbenzene is discussed in detail. The first hydrophosphination step proceeds at ambient temperature without additional promoters, and subsequent addition of base allows full conversion to benzophosphepines. Novel benzeno-1,4-diphosphinanes were isolated as side products. The benzophosphepine complexes themselves serve as convenient phosphinidene precursors at elevated, substituent-dependent temperatures (>55 degrees C). Kinetic and computational analyses support the proposal that the phosphepine-phosphanorcaradiene isomerization is the rate-determining step. In the absence of substrate, addition of the transient phosphinidene to another benzophosphepine molecule is observed, and addition to 1,2-diethynylbenzene furnishes a delicate bidentate diphosphirene complex. PMID- 16316246 TI - Scope and mechanistic study of the ruthenium-catalyzed ortho-C-H bond activation and cyclization reactions of arylamines with terminal alkynes. AB - The cationic ruthenium hydride complex [(PCy(3))(2)(CO)(CH(3)CN)(2)RuH](+)BF(4)( ) was found to be a highly effective catalyst for the C-H bond activation reaction of arylamines and terminal alkynes. The regioselective catalytic synthesis of substituted quinoline and quinoxaline derivatives was achieved from the ortho-C-H bond activation reaction of arylamines and terminal alkynes by using the catalyst Ru(3)(CO)(12)/HBF(4).OEt(2). The normal isotope effect (k(CH)/k(CD) = 2.5) was observed for the reaction of C(6)H(5)NH(2) and C(6)D(5)NH(2) with propyne. A highly negative Hammett value (rho = -4.4) was obtained from the correlation of the relative rates from a series of meta substituted anilines, m-XC(6)H(4)NH(2), with sigma(p) in the presence of Ru(3)(CO)(12)/HBF(4).OEt(2) (3 mol % Ru, 1:3 molar ratio). The deuterium labeling studies from the reactions of both indoline and acyclic arylamines with DCCPh showed that the alkyne C-H bond activation step is reversible. The crossover experiment from the reaction of 1-(2-amino-1-phenyl)pyrrole with DCCPh and HCCC(6)H(4)-p-OMe led to preferential deuterium incorporation to the phenyl substituted quinoline product. A mechanism involving rate-determining ortho-C-H bond activation and intramolecular C-N bond formation steps via an unsaturated cationic ruthenium acetylide complex has been proposed. PMID- 16316247 TI - Critical influence of adsorption geometry in the heterogeneous epoxidation of "allylic" alkenes: structure and reactivity of three phenylpropene isomers on Cu(111). AB - It has long been conjectured that the difficulty of heterogeneously epoxidizing higher alkenes such as propene is due to the presence in the molecule of "allylic" H atoms that are readily stripped off by the oxygenated surface of the metal catalyst resulting in combustion. Here, taking advantage of the intrinsically higher epoxidation selectivity of Cu over Ag under vacuum conditions, we have used three phenylpropene structural isomers to examine the correlation between adsorption geometry and oxidation chemistry. It is found that under comparable conditions alpha-methylstyrene, trans-methylstyrene, and allylbenzene behave very differently on the oxygenated Cu(111) surface: the first undergoes extensive epoxidation accompanied by relatively little decomposition of the alkene; the second leads to some epoxide formation and extensive alkene decomposition; and the third is almost inert with respect to both reaction pathways. This reactive behavior is understandable in terms of the corresponding molecular conformations determined by near-edge X-ray absorption fine structure spectroscopy and density functional theory calculations. The proximity to the surface of the C=C function and of the allylic H atoms is critically important in determining reaction selectivity. This demonstrates the importance of adsorption geometry and confirms that allylic H stripping is indeed a key process that limits epoxidation selectivity in such cases. PMID- 16316248 TI - Enantioselective addition of secondary phosphines to methacrylonitrile: catalysis and mechanism. AB - A highly enantioselective intermolecular hydrophosphination reaction is described. The (Pigiphos)nickel(II)-catalyzed reaction of secondary phosphines and methacrylonitrile gives chiral 2-cyanopropylphosphines in good yield and high enantiomeric excess (ee's up to 94%; (R)-(S)-Pigiphos = bis[(R)-1-[(S)-2 (diphenylphosphino)ferrocenyl]ethyl}cyclohexylphos phine). We propose a mechanism involving coordination of methacrylonitrile to the dicationic nickel catalyst followed by 1,4-addition of the phosphine, and then, rate-determining proton transfer. This mechanism is supported by (a) the experimentally determined rate law (rate = k'[Ni][methacrylonitrile][t-Bu(2)PH]), (b) a large primary deuterium isotope effect k(H)/k(D) = 4.6(1) for the addition of t-Bu(2)PH(D) at 28.3 degrees C in toluene-d(8), (c) the isolation and characterization of the species [Ni(kappa(3)-Pigiphos)(kappaN-methacrylonitrile)](2+), and (d) DFT calculations of model compounds. PMID- 16316249 TI - Light-activated hydrogel formation via the triggered folding and self-assembly of a designed peptide. AB - Photopolymerization can be used to construct materials with precise temporal and spatial resolution. Applications such as tissue engineering, drug delivery, the fabrication of microfluidic devices and the preparation of high-density cell arrays employ hydrogel materials that are often prepared by this technique. Current photopolymerization strategies used to prepare hydrogels employ photoinitiators, many of which are cytotoxic and require large macromolecular precursors that need to be functionalized with moieties capable of undergoing radical cross-linking reactions. We have developed a simple light-activated hydrogelation system that employs a designed peptide whose ability to self assemble into hydrogel material is dependent on its intramolecular folded conformational state. An iterative design strategy afforded MAX7CNB, a photocaged peptide that, when dissolved in aqueous medium, remains unfolded and unable to self-assemble; a 2 wt % solution of freely soluble unfolded peptide is stable to ambient light and has the viscosity of water. Irradiation of the solution (260 < lambda < 360 nm) releases the photocage and triggers peptide folding to produce amphiphilic beta-hairpins that self-assemble into viscoelastic hydrogel material. Circular dichroic (CD) spectroscopy supports this folding and self-assembly mechanism, and oscillatory rheology shows that the resulting hydrogel is mechanically rigid (G' = 1000 Pa). Laser scanning confocal microscopy imaging of NIH 3T3 fibroblasts seeded onto the gel indicates that the gel surface is noncytotoxic, conducive to cell adhesion, and allows cell migration. Lastly, thymidine incorporation assays show that cells seeded onto decaged hydrogel proliferate at a rate equivalent to cells seeded onto a tissue culture-treated polystyrene control surface. PMID- 16316250 TI - Vibrational spectroscopy and structures of Ni+(C2H2)(n) (n =1-4) complexes. AB - Nickel cation-acetylene complexes of the form Ni(+)(C(2)H(2))(n), Ni(+)(C(2)H(2))Ne, and Ni(+)(C(2)H(2))(n)Ar(m) (n = 1-4) are produced in a molecular beam by pulsed laser vaporization. These ions are size-selected and studied in a time-of-flight mass spectrometer by infrared laser photodissociation spectroscopy in the C-H stretch region. The fragmentation patterns indicate that the coordination number is 4 for this system. The n = 1-4 complexes with and without rare gas atoms are also investigated with density functional theory. The combined IR spectra and theory show that pi-complexes are formed for the n = 1-4 species, causing the C-H stretches in the acetylene ligands to shift to lower frequencies. Theory reveals that there are low-lying excited states nearly degenerate with the ground state for all the Ni(+)(C(2)H(2))(n) complexes. Although isomeric structures are identified for rare gas atom binding at different sites, the attachment of rare gas atoms results in only minor perturbations on the structures and spectra for all complexes. Experiment and theory agree that multiple acetylene binding takes place to form low-symmetry structures, presumably due to Jahn-Teller distortion and/or ligand steric effects. The fully coordinated Ni(+)(C(2)H(2))(4) complex has a near-tetrahedral structure. PMID- 16316251 TI - Search for improved host architectures: application of de novo structure-based design and high-throughput screening methods to identify optimal building blocks for multidentate ethers. AB - This paper presents a computational approach to the deliberate design of improved host architectures. The approach, which involves the use of computer-aided design software, is illustrated by application to cation hosts containing multiple aliphatic ether oxygen binding sites. De novo molecule building software, HostDesigner, is interfaced with molecular mechanics software, GMMX, providing a tool for generating and screening millions of potential bidentate building block structures. Enhanced cation binding affinity can be achieved when highly organized building blocks are used to construct macrocyclic hosts. PMID- 16316252 TI - DNA molecular motor driven micromechanical cantilever arrays. AB - The unique ability of living systems to translate biochemical reactions into mechanical work has inspired the design of synthetic DNA motors which generate nanoscale motion via controlled conformational change. However, while Nature has evolved intricate mechanisms to convert molecular shape change into specific micrometer-scale mechanical cellular responses, the integration of artificial DNA motors with mechanical devices presents a major challenge. Here we report the direct integration between an ensemble of DNA motors and an array of microfabricated silicon cantilevers. The forces exerted by the precise duplex to nonclassical i-motif conformational change were probed via differential measurements using an in-situ reference cantilever coated with a nonspecific sequence of DNA. Fueled by the addition of protons, the open to close stroke of the motor induced 32 +/- 3 mN/m compressive surface stress, which corresponds to a single motor force of approximately 11 pN/m, an order of magnitude larger than previous classical hybridization studies. Furthermore, the surface-tethered conformational change was found to be highly reversible, in contrast to classical DNA motors which typically suffer rapid system poisoning. The direction and amplitude of motor-induced cantilever motion was tuneable via control of buffer pH and ionic strength, indicating that electrostatic forces play an important role in stress generation. Hybrid devices which directly harness the multiple accessible conformational states of dynamic oligonucleotides and aptamers, translating biochemical energy into micromechanical work, present a radical new approach to the construction of "smart" nanoscale machinery and mechano biosensors. PMID- 16316253 TI - Strong solute-solute dispersive interactions in a protein-ligand complex. AB - The contributions of solute-solute dispersion interactions to binding thermodynamics have generally been thought to be small, due to the surmised equality between solute-solvent dispersion interactions prior to the interaction versus solute-solute dispersion interactions following the interaction. The thermodynamics of binding of primary alcohols to the major urinary protein (MUP I) indicate that this general assumption is not justified. The enthalpy of binding becomes more favorable with increasing chain length, whereas the entropy of binding becomes less favorable, both parameters showing a linear dependence. Despite the hydrophobicity of the interacting species, these data show that binding is not dominated by the classical hydrophobic effect, but can be attributed to favorable ligand-protein dispersion interactions. PMID- 16316254 TI - Mechanistic insights into an unprecedented C-C bond activation on a Rh/Ga bimetallic complex: a combined experimental/computational approach. AB - The unusual rearrangement of [RhCp(GaCp)(CH(3))(2)] (1c) to [RhCp(C(5)Me(4)Ga(CH(3))(3))] (2) is presented and its mechanism is discussed in detail. (13)C MAS NMR spectroscopy revealed that the title reaction proceeds cleanly not only in solution but also in solid state, which supports a unimolecular reaction pathway. On the basis of (1)H, (13)C, and ROESY NMR spectroscopy as well as isolation and structural elucidation of the hydrolysis product, the compound [RhCp(endo-eta(4)-C(5)Me(5)GaMe(2))] (3a) was identified as a crucial reaction intermediate. DFT calculations on the B3LYP level of theory support this assignment and suggest a concerted C-C bond activation mechanism that topologically takes place at the gallium center. Furthermore, two fluxional processes of the reaction intermediate 3a were studied experimentally as well as by computational methods. First, a mechanism takes place similar to a ring slipping process that exchanges a GaMe(2) group between adjacent ring carbon atoms within the same Cp ring. This process proceeds at a rate comparable to the NMR time scale and indeed is calculated to be energetically very favorable. Second, a unimolecular exchange process of the GaMe(2) group between the two Cp rings of 3a could be experimentally proven by the introduction of phenyl substituents as a label into the Cp ligands at both sites, the rhodium as well as the gallium center. A series of experiments including deuteration studies and competition reactions was performed to substantiate the suggested mechanism being in accordance with DFT calculations on possible transition states. PMID- 16316255 TI - A complete set of NMR chemical shifts and spin-spin coupling constants for L Alanyl-L-alanine zwitterion and analysis of its conformational behavior. AB - With the aid of labeling with stable isotopes ((15)N and (13)C) a complete set of chemical shifts and indirect spin-spin coupling constants was obtained for the zwitterionic form of L-alanyl-L-alanine in aqueous solution. Different sensitivities of the NMR parameters to the molecular geometry were discussed on the basis of comparison with ab initio (DFT) calculated values. An adiabatic two dimensional vibrational wave function was constructed and used for determination of the main chain torsion angle dispersions and conformational averaging of the NMR shifts and coupling constants. The quantum description of the conformational dynamics based on the density functional theory and a polarizable continuum solvent model agrees reasonably with classical molecular dynamics simulations using explicit solvent. The results consistently evidence the presence of a single form in the aqueous solution with equilibrium main chain torsion angle values (psi = 147 degrees, varphi = -153 degrees), close to that one found previously in an X-ray study. Under normal temperature the torsion angles can vary by about 10 degrees around their equilibrium values, which leads, however, to minor corrections of the NMR parameters only. The main chain heavy atom chemical shifts and spin-spin coupling constants involving the alpha-carbon and hydrogen atoms appear to be most useful for the peptide structural predictions. PMID- 16316256 TI - Three-dimensional self-organization of supramolecular self-assembled porphyrin hollow hexagonal nanoprisms. AB - A self-assembly technique assisted with surfactant is developed to fabricate one dimensional (1D) nanostructure of zinc meso-tetra (4-pyridyl) porphyrin. The so prepared nanostructure appears in a shape of hollow hexagonal nanoprism with uniform size. The length and aspect ratio of the nanoprisms is easily tunable by controlling the stoichiometric ratio of porphyrin over surfactant. The internal structure of the nanoprisms is well described by XRD. Furthermore, as a result of dispersivity and regular geometric shape, these nanoprisms can readily self organize into an ordered, smectic three-dimensional (3D) architecture through simple evaporation of the solvent. The results should be significant in porphyrin crystallization and porphyrin application in optoelectronic device, catalysis, drug delivery, and molecular filtration. PMID- 16316257 TI - Weak intramolecular interactions in ethylene glycol identified by vapor phase OH stretching overtone spectroscopy. AB - Vapor phase OH-stretching overtone spectra of ethylene glycol were recorded to investigate weak intramolecular hydrogen bonding. The spectra were recorded with conventional absorption spectroscopy and laser photoacoustic spectroscopy in the first to fourth OH-stretching overtone regions. The room-temperature spectra are dominated by two conformers that show weak intramolecular hydrogen bonding. A less abundant third conformer, with no sign of hydrogen bonding, is also observed. Vapor phase spectra of the ethylene-d(4) glycol isotopomer were also recorded and used to identify an interfering resonance between CH-stretching and OH-stretching states in the fourth overtone. Anharmonic oscillator local mode calculations of the OH-stretching transitions have provided an accurate simulation of the observed spectra. The local mode parameters were calculated with coupled cluster ab initio methods. The calculations facilitate assignment of the different conformers in the spectra and illustrate the effect of the intramolecular hydrogen bonding. PMID- 16316258 TI - Kinetic capillary electrophoresis (KCE): a conceptual platform for kinetic homogeneous affinity methods. AB - We propose kinetic capillary electrophoresis (KCE) as a conceptual platform for the development of kinetic homogeneous affinity methods. KCE is defined as the CE separation of species that interact during electrophoresis. Depending on how the interaction is arranged, different KCE methods can be designed. All KCE methods are described by the same mathematics: the same system of partial differential equations with only initial and boundary conditions being different. Every qualitatively unique set of initial and boundary conditions defines a unique KCE method. Here, we (i) present the theoretical bases of KCE, (ii) define four new KCE methods, and (iii) propose a multimethod KCE toolbox as an integrated kinetic technique. Using the KCE toolbox, we were able to, for the first time, observe high-affinity (specific) and low-affinity (nonspecific) interactions within the same protein-ligand pair. The concept of KCE allows for the creation of an expanding toolset of powerful kinetic homogeneous affinity methods, which will find their applications in studies of biomolecular interactions, quantitative analyses, and selecting affinity probes and drug candidates from complex mixtures. PMID- 16316259 TI - Catalyst-controlled asymmetric synthesis of fostriecin and 8-epi-fostriecin. AB - Catalytic asymmetric synthesis of the natural antibiotic fostriecin (CI-920) and its analogue 8-epi-fostriecin and evaluation of their biological activity are described. We used four catalytic asymmetric reactions to construct all of the chiral centers of fostriecin and 8-epi-fostriecin; cyanosilylation of a ketone, Yamamoto allylation, direct aldol reaction, and Noyori reduction, two of which were developed by our group. Catalytic enantioselective cyanosilylation of ketone 13 produced the chiral tetrasubstituted carbon at C-8. Both enantiomers of the product cyanohydrin were obtained with high enantioselectivity by switching the center metal of the catalyst from titanium to gadolinium. Yamamoto allylation constructed the C-5 chiral carbon in the alpha,beta-unsaturated lactone moiety. A direct catalytic asymmetric aldol reaction of an alkynyl ketone using LLB catalyst constructed the chirality at C-9 with the introduction of a synthetically versatile alkyne moiety, which was later converted to cis-vinyl iodide, the substrate for the subsequent Stille coupling for the triene synthesis. Noyori reduction produced the secondary alcohol at C-11 from the acetylene ketone 6 with excellent selectivity. Importantly, all the stereocenters were constructed under catalyst control in this synthesis. This strategy should be useful for rapid synthesis of stereoisomers of fostriecin. PMID- 16316260 TI - Kinetically controlled synthesis of triangular and hexagonal nanoplates of palladium and their SPR/SERS properties. AB - The rapid reduction of Na(2)PdCl(4) by ethylene glycol in the presence of poly(vinyl pyrrolidone) (PVP) has recently been demonstrated as a convenient method of generating Pd cubooctahedra and twinned nanoparticles. Here we describe a new procedure where Pd triangular or hexagonal nanoplates could be selectively synthesized by manipulating the reduction kinetics of the polyol process. More specifically, the reduction rate was substantially reduced through the introduction of Fe(III) species and the O(2)/Cl(-) pair, two wet etchants for Pd(0). The etching power of the O(2)/Cl(-) pair could be further enhanced by adding an acid to lower the pH of the reaction solution. Unlike the previously reported synthesis of Ag and Au nanoplates, light was found to have no indispensable role in the formation of Pd nanoplates. Both triangular and hexagonal nanoplates of Pd exhibited surface plasmon resonance (SPR) peaks in the visible region, and their positions matched with the results of discrete dipole approximation (DDA) calculation. Thanks to their sharp corners and edges, these Pd nanoplates could serve as active substrates for surface-enhanced Raman scattering (SERS). PMID- 16316263 TI - Special section on theoretical models and conceptual analyses: second installment. AB - The Journal of Applied Psychology's call for theoretical models and conceptual analyses brought a terrific response. The first set of articles accepted in response to the call appeared in the December 2004 issue. This installment contains the second set of articles. PMID- 16316264 TI - Adaptation-level theory, opponent process theory, and dispositions: an integrated approach to the stability of job satisfaction. AB - Research suggests that the stability of job satisfaction is partially the result of dispositions (J. J. Connolly & C. Viswesvaran, 2000; C. Dormann & D. Zapf, 2001; T. A. Judge & J. E. Bono, 2001a; T. A. Judge, D. Heller, & M. K. Mount, 2002). Opponent process theory (R. L. Solomon & J. D. Corbit, 1973, 1974) and adaptation-level theory (H. Helson, 1948) are alternative explanations of this stability that explain how environmental effects on job satisfaction dissipate across time. On the basis of an integration of these explanations, the authors propose that dispositions (a) influence employees' equilibrium or adaptation level of job satisfaction, (b) influence employees' sensitivity to workplace events, and (c) influence the speed at which job satisfaction returns to equilibrium after one is exposed to a workplace event. Research and applied implications are discussed. PMID- 16316265 TI - An episodic process model of affective influences on performance. AB - In this article, the authors present a model linking immediate affective experiences to within-person performance. First, the authors define a time structure for performance (the performance episode) that is commensurate with the dynamic nature of affect. Next, the authors examine the core cognitive and regulatory processes that determine performance for 1 person during any particular episode. Third, the authors describe how various emotions and moods influence the intermediary performance processes, thereby affecting performance. In the final section of the article, the authors discuss limitations, future research directions, and practical implications for their episodic process model of affect and performance. PMID- 16316266 TI - Accounting for subordinate perceptions of supervisor power: an identity dependence model. AB - The authors present a model that explains how subordinates perceive the power of their supervisors and the causal mechanisms by which these perceptions translate into subordinate outcomes. Drawing on identity and resource-dependence theories, the authors propose that supervisors have power over their subordinates when they control resources needed for the subordinates' enactment and maintenance of current and desired identities. The joint effect of perceptions of supervisor power and supervisor intentions to provide such resources leads to 4 conditions ranging from highly functional to highly dysfunctional: confirmation, hope, apathy, and progressive withdrawal. Each of these conditions is associated with specific outcomes such as the quality of the supervisor-subordinate relationship, turnover, and changes in the type and centrality of various subordinate identities. PMID- 16316267 TI - Understanding the emergence of state goal orientation in organizational work groups: the role of leadership and multilevel climate perceptions. AB - This article attends to a broad range of practically significant employee motivations and provides insight into how to enhance individual-level performance by examining individual-level state goal orientation emergence in organizational work groups. Leadership and multilevel climate processes are theorized to parallel each dimension of state goal orientation to cue and ultimately induce the corresponding achievement focus among individual work group members. It is argued that the patterns of leader behavior, which elucidate the leader's achievement priority, shape group members' psychological and work group climate to embody this priority. Resulting multilevel climate perceptions signal and compel group members to adopt the ascribed form of state goal orientation. The quality of the leader-member exchange relationship is viewed as a means to clarify leader messages in the formation of group members' psychological climate and internalize these cues in the emergence of state goal orientation. Considerations for future research and practice are discussed. PMID- 16316268 TI - A motivated action theory account of goal orientation. AB - Rapid organizational change is increasing the pressure on employees to continually update their skills and adapt their behavior to new organizational realities. Goal orientation is a promising motivational construct that may explain why some individuals adapt to change better. Unfortunately, the current goal orientation literature is in a state of conceptual and methodological disarray. This presentation reviews the goal orientation literature and identifies numerous conceptual ambiguities, including definitional inconsistencies, dimensional inconsistencies, and inconsistencies in the conceptualization of stability. These conceptual ambiguities result in a confusing array of goal orientation measures and manipulations and ultimately an incoherent empirical database. A dynamic self-regulation model of goal orientation, termed motivated action theory, is presented to integrate the various conceptual perspectives and to provide guidelines for future goal orientation research. PMID- 16316269 TI - The cultural mosaic: a metatheory for understanding the complexity of culture. AB - Workforce population trends have increased the numbers and kinds of culturally diverse people who work together. Researchers in organizational behavior have often examined culture through values; however, cultural values can be based on collections of people other than traditional nation states. A cultural mosaic is presented as a framework to identify demographic, geographic, and associative features underlying culture. An individual's unique collage of multiple cultural identities yields a complex picture of the cultural influences on that person. Developments in chaos and complexity theories are proposed as a theoretical base for study on the complexity of culture at the individual level. Additional developments in network theory serve as a theoretical base for cultural research at the group level. The cultural mosaic is described as a complex system with localized structures, linking cultural tiles in ordered and chaotic ways. Research propositions examining multiple cultural identities at individual and group levels are discussed. PMID- 16316270 TI - The effects of sexual harassment on turnover in the military: time-dependent modeling. AB - Sexual harassment has consistently negative consequences for working women, including changes in job attitudes (e.g., lower satisfaction) and behaviors (e.g., increased work withdrawal). Cross-sectional evidence suggests that harassment influences turnover intentions. However, few studies have used actual turnover; rather, they rely on proxies. With a sample of 11,521 military servicewomen with turnover data spanning approximately 4 years, the authors used the appropriate method for longitudinal turnover data--Cox's regression--to investigate the impact of harassment on actual turnover. Experiences of harassment led to increased turnover, even after controlling for job satisfaction, organizational commitment, and marital status. Among officers, harassment also affected turnover over and above rank. Given turnover's relevance to organizational bottom lines, these findings have important implications not only for individual women but also for organizations. PMID- 16316271 TI - Adaptation of teams in response to unforeseen change: effects of goal difficulty and team composition in terms of cognitive ability and goal orientation. AB - Halfway through a 3-hour experiment in which 64 3-person teams needed to make a series of decisions, a communications channel began to deteriorate, and teams needed to adapt their system of roles in order to perform effectively. Consistent with previous research, team composition with respect to members' cognitive ability was positively associated with adaptation. Adaptation was also influenced by interactions of team goal difficulty and team composition with respect to team members' goal orientation. Teams with difficult goals and staffed with high performance orientation members were especially unlikely to adapt. Teams with difficult goals and staffed with high-learning orientation members were especially likely to adapt. Supplemental analyses provided insight into the observed effects in that the difficulty of team goals and members' goal orientation predicted interpersonal, transition, and action processes, all of which predicted team adaptation. PMID- 16316272 TI - Organizational justice and Black applicants' reactions to affirmative action. AB - Using organizational justice as a guiding framework, the authors studied perceptions of affirmative action programs by presumed beneficiaries. Three conceptual issues were addressed: (a) the content of different affirmative action plans; (b) the 3-way interaction among distributive, procedural, and interactional justice; and (c) the distinction between outcome favorability and distributive justice. These ideas were tested with a sample of Black engineering students who responded to 1 of 6 plans. Participants distinguished among the various plans, with some policies being viewed as more fair than others. In addition, a 3-way interaction among the 3 types of organizational justice was observed. Specifically, the 2-way interaction between distributive and interactional fairness was only significant when procedural justice was low. Implications for organizational justice and for the design of affirmative action programs are discussed. PMID- 16316273 TI - The Great Eight competencies: a criterion-centric approach to validation. AB - The author presents results of a meta-analysis of 29 validation studies (N=4,861) that uses the Great Eight competency factors (Kurz & Bartram, 2002) as the criterion measurement framework. Predictors of the Great Eight competencies based only on personality scales show moderate to good correlations with line-manager ratings for all 8 of the competencies. On their own, ability tests correlate with 4 of the 8 competencies, and together ability and personality data yield operational validities ranging from 0.20 to 0.44 for the 8 competencies. Operational validities for aggregated predictors with aggregated criteria were estimated to be 0.53. The value of differentiating the criterion space and of relating predictor variables to criterion variables in a one-to-one fashion is discussed. PMID- 16316274 TI - In search of the elusive U-shaped performance-turnover relationship: are high performing Swiss bankers more liable to quit? AB - This project revisits the perennial debate over the relationship between job performance and turnover. Disputing traditional findings, C. Trevor, B. Gerhart, and J. Boudreau (1997) observed that high and low performers quit more than do average performers. They further challenged received wisdom by showing that promotions can induce turnover, especially among poor performers, by signaling ability. The authors sought to replicate and extend these unconventional findings by exploring curvilinear and moderating effects on the performance-exit relationship among 11,098 Swiss nationals employed in a bank. Survival regression revealed that performance is curvilinearly related to quits and that bonus pay deterred superior performers from leaving more than did pay increases. Further, the average number of job levels advanced per promotion rather than promotion rate increased quit risks. Cultural and organizational moderators of performance termination associations and effective strategies for retaining top performers are discussed. PMID- 16316275 TI - Linking organizational resources and work engagement to employee performance and customer loyalty: the mediation of service climate. AB - This study examined the mediating role of service climate in the prediction of employee performance and customer loyalty. Contact employees (N=342) from 114 service units (58 hotel front desks and 56 restaurants) provided information about organizational resources, engagement, and service climate. Furthermore, customers (N=1,140) from these units provided information on employee performance and customer loyalty. Structural equation modeling analyses were consistent with a full mediation model in which organizational resources and work engagement predict service climate, which in turn predicts employee performance and then customer loyalty. Further analyses revealed a potential reciprocal effect between service climate and customer loyalty. Implications of the study are discussed, together with limitations and suggestions for future research. PMID- 16316276 TI - Organizational error management culture and its impact on performance: a two study replication. AB - The authors argue that a high-organizational error management culture, conceptualized to include norms and common practices in organizations (e.g., communicating about errors, detecting, analyzing, and correcting errors quickly), is pivotal to the reduction of negative and the promotion of positive error consequences. Organizational error management culture was positively related to firm performance across 2 studies conducted in 2 different European countries. On the basis of quantitative and qualitative cross-sectional data from 65 Dutch organizations, Study 1 revealed that organizational error management culture was significantly correlated with both organizational goal achievement and an objective indicator of economic performance. This finding was confirmed in Study 2, using change-of-profitability data from 47 German organizations. The results suggest that organizations may want to introduce organizational error management as a way to boost firm performance. PMID- 16316277 TI - A meta-analysis of the relationship between organizational citizenship behavior and counterproductive work behavior. AB - Job performance is increasingly being seen to encompass constructs such as organizational citizenship behavior (OCB) and counterproductive work behavior (CWB). To clarify the OCB-CWB relationship, a meta-analysis was conducted. Results indicate a modest negative relationship (p = -0.32). The relationship strength did not increase appreciably when the target of the behavior (the organization vs. other employees) was the same. Moreover, OCB and CWB exhibited somewhat distinct patterns of relationships with antecedents. The OCB-CWB relationship was moderated by the source of the ratings, the presence of antithetical items, and the type of response options. An employee-centric perspective is proposed whereby both OCB and CWB are perceived as adaptive behavior. Implications for organizations are discussed. PMID- 16316278 TI - Emotional display rules and emotional labor: the moderating role of commitment. AB - The authors examined whether commitment to emotional display rules is a necessary condition for emotional display rules to affect behavior at work. Results using structural equation modeling revealed that display rule commitment moderated the relationships of emotional display rule perceptions with surface acting, deep acting, and positive affective delivery at work, such that the relationships were strong and positive when commitment to display rules was high and weak when commitment to display rules was low. These findings suggest that motivation plays a role in the emotional labor process in that individuals must be committed to display rules for these rules to affect behavior. PMID- 16316279 TI - The mediating role of self-efficacy in the development of entrepreneurial intentions. AB - The purpose of this study was to investigate the mediating role of self-efficacy in the development of students' intentions to become entrepreneurs. The authors used structural equation modeling with a sample of 265 master of business administration students across 5 universities to test their hypotheses. The results showed that the effects of perceived learning from entrepreneurship related courses, previous entrepreneurial experience, and risk propensity on entrepreneurial intentions were fully mediated by entrepreneurial self-efficacy. Contrary to expectations, gender was not mediated by self-efficacy but had a direct effect such that women reported lower entrepreneurial career intentions. The authors discuss practical implications and directions for future research. PMID- 16316280 TI - The dynamic spillover of satisfaction between work and marriage: the role of time and mood. AB - Previous research has indicated important linkages between work and family domains and roles. However, the nature of the dynamic spillover between job and marital satisfaction remains poorly understood. The current study tests both the concurrent and lagged associations between job and marital satisfaction at a within-individual level of analysis using a diary study of 76 fully employed, married adults. The authors further examine the mediating role of mood in this spillover process. Consistent with their hypotheses, findings indicate both a concurrent and a lagged (job to marital and marital to job) job satisfaction marital satisfaction association at the within-subject level of analysis and lend some support for the mediating role of mood (most notably positive affect) in these associations. The authors hope these findings stimulate new research that uses more complex designs and comprehensive theoretical models to investigate work-family links. PMID- 16316281 TI - Performance differences among four organizational commitment profiles. AB - The authors drew from prior research on organizational commitment and from configural organizational theory to propose a framework of affective and continuance commitment profiles. Using cluster analyses, the authors obtained evidence for 4 of these profiles in an energy industry sample (N=970) and a sample of 345 employed college students. The authors labeled the clusters: allied (i.e., moderate affective and continuance commitment), free agents (moderate continuance commitment and low affective commitment), devoted (high affective and continuance commitment), and complacent (moderate affective and low continuance commitment). Using a subset of the employed student sample (n=148), the authors also found that the free agents received significantly poorer supervisor ratings of performance, organizational citizenship behavior, and antisocial behavior than any other group. PMID- 16316282 TI - Contingent leadership and effectiveness of trauma resuscitation teams. AB - This research investigated leadership and effectiveness of teams operating in a high-velocity environment, specifically trauma resuscitation teams. On the basis of the literature and their own ethnographic work, the authors proposed and tested a contingency model in which the influence of leadership on team effectiveness during trauma resuscitation differs according to the situation. Results indicated that empowering leadership was more effective when trauma severity was low and when team experience was high. Directive leadership was more effective when trauma severity was high or when the team was inexperienced. Findings also suggested that an empowering leader provided more learning opportunities than did a directive leader. The major contribution of this article is the linkage of leadership to team effectiveness, as moderated by relatively specific situational contingencies. PMID- 16316283 TI - Intentions to work during terminal illness: an exploratory study of antecedent conditions. AB - Facing a terminal illness is an unimaginably difficult experience, yet many individuals intend to work despite their prognosis. However, research has not systematically examined the potential antecedents underlying such intentions. Using behavioral intention theory as an underlying framework, this study hypothesized that reasons for working (intrinsic and extrinsic), the will to live, disability severity, accessibility of travel, and age would predict intentions to work during terminal illness. A representative sample of medically diagnosed amyotrophic lateral sclerosis (a.k.a. Lou Gehrig's disease) patients with a mean life expectancy of approximately 3 years participated (mean age=57.8 years). Controlling for length of diagnosis, employment status, and demographic variables, results indicated that intrinsic reasons were particularly strong predictors of intentions, followed by age, disability severity, and accessibility of travel. Exploratory findings also indicated that behavioral intentions were positively related to future employment status, consistent with past theory. ((c) 2005 APA, all rights reserved). PMID- 16316284 TI - The advice and influence networks of transformational leaders. AB - Existing transformational leadership research has focused primarily on the behaviors of leaders and their effects on followers. The authors extended this research by examining the social networks of managers who exhibit transformational leadership behaviors. Their focus was on the network of relationships that managers develop and whether they hold key positions in the organization's informal social networks. In a field study using data from 39 managers and 130 nonmanagement employees of 6 organizations, the authors found that managers who score higher on transformational leadership tend to hold more central positions in organizational advice and influence networks. Furthermore, the direct reports of these leaders were also more central in informal organizational networks. These results illuminate one of the ways that managers who exhibit transformational leadership behaviors may exert influence in organizations. PMID- 16316285 TI - Visual search and the collapse of categorization. AB - Categorization researchers typically present single objects to be categorized. But real-world categorization often involves object recognition within complex scenes. It is unknown how the processes of categorization stand up to visual complexity or why they fail facing it. The authors filled this research gap by blending the categorization and visual-search paradigms into a visual-search and categorization task in which participants searched for members of target categories in complex displays. Participants have enormous difficulty in this task. Despite intensive and ongoing category training, they detect targets at near-chance levels unless displays are extremely simple or target categories extremely focused. These results, discussed from the perspectives of categorization and visual search, might illuminate societally important instances of visual search (e.g., diagnostic medical screening). PMID- 16316286 TI - How does attention select and track spatially extended objects? New effects of attentional concentration and amplification. AB - Real-world situations involve attending to spatially extended objects, often under conditions of motion and high processing load. The present experiments investigated such processing by requiring observers to attentionally track a number of long, moving lines. Concurrently, observers responded to sporadic probes as a measure of the distribution of attention across the lines. The results revealed that attention is concentrated at the centers of lines during tracking, despite their uniformity, and that this center advantage grew as the lines became longer: Not only did observers get worse near the endpoints, but they became better at the lines' centers, as if attention became more concentrated as the objects became more extended. These results begin to show how attention is flexibly allocated in online visual processing to extended dynamic objects. PMID- 16316287 TI - Integration of multiple speech segmentation cues: a hierarchical framework. AB - A central question in psycholinguistic research is how listeners isolate words from connected speech despite the paucity of clear word-boundary cues in the signal. A large body of empirical evidence indicates that word segmentation is promoted by both lexical (knowledge-derived) and sublexical (signal-derived) cues. However, an account of how these cues operate in combination or in conflict is lacking. The present study fills this gap by assessing speech segmentation when cues are systematically pitted against each other. The results demonstrate that listeners do not assign the same power to all segmentation cues; rather, cues are hierarchically integrated, with descending weights allocated to lexical, segmental, and prosodic cues. Lower level cues drive segmentation when the interpretive conditions are altered by a lack of contextual and lexical information or by white noise. Taken together, the results call for an integrated, hierarchical, and signal-contingent approach to speech segmentation. PMID- 16316288 TI - Grammatical gender effects on cognition: implications for language learning and language use. AB - In 4 experiments, the authors addressed the mechanisms by which grammatical gender (in Italian and German) may come to affect meaning. In Experiments 1 (similarity judgments) and 2 (semantic substitution errors), the authors found Italian gender effects for animals but not for artifacts; Experiment 3 revealed no comparable effects in German. These results suggest that gender effects arise as a generalization from an established association between gender of nouns and sex of human referents, extending to nouns referring to sexuated entities. Across languages, such effects are found when the language allows for easy mapping between gender of nouns and sex of human referents (Italian) but not when the mapping is less transparent (German). A final experiment provided further constraints: These effects during processing arise at a lexical-semantic level rather than at a conceptual level. PMID- 16316289 TI - Encoding multielement scenes: statistical learning of visual feature hierarchies. AB - The authors investigated how human adults encode and remember parts of multielement scenes composed of recursively embedded visual shape combinations. The authors found that shape combinations that are parts of larger configurations are less well remembered than shape combinations of the same kind that are not embedded. Combined with basic mechanisms of statistical learning, this embeddedness constraint enables the development of complex new features for acquiring internal representations efficiently without being computationally intractable. The resulting representations also encode parts and wholes by chunking the visual input into components according to the statistical coherence of their constituents. These results suggest that a bootstrapping approach of constrained statistical learning offers a unified framework for investigating the formation of different internal representations in pattern and scene perception. PMID- 16316290 TI - How subjective grouping of options influences choice and allocation: diversification bias and the phenomenon of partition dependence. AB - The authors argue that people's tendency to diversify their allocations of money and consumption choices over alternatives gives rise to decisions that vary systematically with the subjective grouping of available options. These subjective groupings are influenced by subtle variations in the presentation of options or elicitation of preferences. Studies 1-4 demonstrate such "partition dependence" in allocations of money to beneficiaries, consumption experiences to future time periods, and choices to a menu of consumption options. Study 5 documents weaker partition dependence among individuals with greater relevant experience discriminating among options, and Study 6 shows that the effect is attenuated among participants with stronger or more accessible intrinsic preferences. PMID- 16316291 TI - The automaticity of visual statistical learning. AB - The visual environment contains massive amounts of information involving the relations between objects in space and time, and recent studies of visual statistical learning (VSL) have suggested that this information can be automatically extracted by the visual system. The experiments reported in this article explore the automaticity of VSL in several ways, using both explicit familiarity and implicit response-time measures. The results demonstrate that (a) the input to VSL is gated by selective attention, (b) VSL is nevertheless an implicit process because it operates during a cover task and without awareness of the underlying statistical patterns, and (c) VSL constructs abstracted representations that are then invariant to changes in extraneous surface features. These results fuel the conclusion that VSL both is and is not automatic: It requires attention to select the relevant population of stimuli, but the resulting learning then occurs without intent or awareness. PMID- 16316292 TI - Moderators of the relationship between implicit and explicit evaluation. AB - Automatic and controlled modes of evaluation sometimes provide conflicting reports of the quality of social objects. This article presents evidence for 4 moderators of the relationship between automatic (implicit) and controlled (explicit) evaluations. Implicit and explicit preferences were measured for a variety of object pairs using a large sample. The average correlation was r=.36, and 52 of the 57 object pairs showed a significant positive correlation. Results of multilevel modeling analyses suggested that (a) implicit and explicit preferences are related, (b) the relationship varies as a function of the objects assessed, and (c) at least 4 variables moderate the relationship: self presentation, evaluative strength, dimensionality, and distinctiveness. The variables moderated implicit-explicit correspondence across individuals and accounted for much of the observed variation across content domains. The resulting model of the relationship between automatic and controlled evaluative processes is grounded in personal experience with the targets of evaluation. PMID- 16316293 TI - Putting some feeling into it--the conceptual and empirical relationships between the classic and emotional Stroop tasks: comment on Algom, Chajut, and Lev (2004). AB - D. Algom, E. Chajut, and S. Lev presented a series of definitional, conceptual, and empirical arguments in support of their conclusion that the classic and emotional Stroop effects are, in their words, "unrelated phenomena" (p. 336), such that the term emotional Stroop effect is a misnomer in reference to the relatively greater interference in ink color naming of emotional versus neutral words. These are strong claims. In this comment, the author critically examines each component of Algom et al.'s case and argues that, in fact, none of these components represents compelling evidence in support of their eventual conclusions. PMID- 16316295 TI - Relations among categorization, induction, recognition, and similarity: comment on Sloutsky and Fisher (2004). AB - V. M. Sloutsky and A. V. Fisher reported 5 experiments documenting relations among categorization, induction, recognition, and similarity in children as well as adults and proposed a new model of induction, SINC (similarity, induction, categorization). Those authors concluded that induction depends on perceptual similarity rather than conceptual knowledge. Despite the useful contributions of this work, there are some important limitations. The experimental designs examined a limited range of phenomena that are not the most revealing about the use of nonperceptual information. The main results involved a simple triad task, for which the SINC model's predictions are equivalent to the predictions of previous models of inductive reasoning. It is also unclear whether the SINC model can account for the observed relations between similarity and recognition. Implications for future work on induction and related cognitive activities are discussed. PMID- 16316297 TI - Pharmacogenomics in cardiovascular disease: the role of single nucleotide polymorphisms in improving drug therapy. AB - Pharmacogenomics is the study of how an individual's genetic inheritance affects the body's response to drugs. Pharmacogenomics holds the promise that drugs might one day be tailor-made for individuals and adapted to an individual's genetic makeup. Several studies have shown that both adverse and beneficial responses to cardiovascular drugs can be influenced by single nucleotide polymorphisms in genes coding for metabolising enzymes, drug transporters and drug targets. Despite the large amount of data about gene-drug interactions, the translation of pharmacogenomics in clinical practise is slow. To improve this, there is a need of new technology and large prospective trials allowing for simultaneous analysis of multiple genetic variants in molecular pathways that could affect drug disposition and action. PMID- 16316298 TI - The PPAR genes, cardiovascular disease and the emergence of PPAR pharmacogenetics. AB - The peroxisome proliferator-activated receptors (PPARs) comprise a group of related transcription factors that serve to regulate a number of cellular processes that are central to cardiovascular health and disease. Two large bodies of work strongly implicate the PPARs as key factors in normal cardiovascular physiology and in cardiovascular pathophysiology: i) studies demonstrating associations between PPAR and abnormal cardiovascular phenotypes; and ii) pharmacological studies assessing the effects of specific PPAR agonists in clinical trials. With the abundance of data available from these studies as a background, PPAR pharmacogenetics has become a promising and rapidly-advancing field. This review summarises the current state of understanding of PPAR pharmacogenetics and its profound implications for the individualisation of therapy for patients with a diverse group of cardiovascular diseases. PMID- 16316299 TI - Does pharmacogenetics have the potential to allow the individualisation of immunosuppressive drug dosing in organ transplantation? AB - The immunosuppressive drugs used in organ transplantation have a narrow therapeutic index, with rejection occurring as a consequence of underdosing and infection, malignancy and a number of drug-specific side effects with excessive dosing. Significant heterogeneity in the dose of drug required to achieve therapeutic blood concentrations adds to the complexity of the problem, which has been partly resolved by therapeutic drug monitoring. Single nucleotide polymorphisms have been identified in genes encoding metabolic enzymes, drug efflux pumps and drug targets for most of the drugs in widespread use. A pharmacogenetic approach to immunosuppressive drug prescribing remains to be tested. Based on current evidence, the most promising strategy would be use of the cytochrome P450 3A5 expressor genotype to guide initial dosing with tacrolimus. PMID- 16316300 TI - Pharmacogenetics of colorectal cancer. AB - Colorectal cancer is one of the most common types of cancer in both men and women. Multiple chemotherapy combinations exist; however, there is currently no strategy for individualised therapy selection prior to treatment. Genetic polymorphisms in genes involved in the metabolism, transport or targets for the commonly used chemotherapy drugs (5-fluorouracil, irinotecan and oxaliplatin) have been described. Many require validation in large prospective trials before they can be used as markers for outcome and/or toxicity. This review describes the data available on polymorphisms in key genes that are associated with chemotherapy toxicity and response in colorectal cancer. PMID- 16316301 TI - Pharmacoeconomics of antifungal pharmacotherapy--challenges and future directions. AB - The frequency and severity of invasive fungal infections have been increasingly recognised and new antifungal therapies have expanded the therapeutic armamentarium available to manage such infections. Antifungal agents comprise a significant portion of antibiotic expenditures at major medical centres, prompting adoption of cost-containment measures and treatment guidelines. This paper reviews available data regarding the costs associated with managing fungal infections, including pharmacoeconomic analyses that have been performed in the setting of documented fungal infections, as well as prophylactic and empiric use of antifungal agents. The challenges of performing such studies are discussed, as well as the limitations of published investigations. Finally, recommendations are made regarding the design and implementation of future pharmacoeconomic analyses that can help establish the true costs of managing invasive fungal infections in at-risk patient populations. PMID- 16316302 TI - Cryptococcal disease and HIV infection. AB - In the era of highly active antiretroviral therapy for the treatment of HIV infection, the dramatic reductions in mortality and morbidity associated with immune reconstitution have included a marked decline in the incidence of opportunistic infections. Cryptococcus neoformans is a yeast that causes predominantly neurological disease in immunocompromised individuals, in particular those with HIV infection. It continues to be an important diagnosis in developing areas and amongst late presenters in parts of the world with access to highly active antiretroviral therapy. This article reviews the epidemiology, clinical features and management of cryptococcal disease in HIV-infected patients, particularly focusing on the history of, current guidelines for and future developments in antifungal therapy. PMID- 16316303 TI - Effective measures for controlling trypanosomiasis. AB - African trypanosomiasis, otherwise known as sleeping sickness in humans and 'Nagana' in cattle, is a disease that is resurgent in Africa. Research on the disease suggests that the development of a vaccine is still far away; even existing drugs are becoming ineffective on account of the emergence of drug resistant trypanosomes. All this contributes to heavy economic losses and a sociopolitical crisis in the continent, thus underscoring the pressure to intensify research for inexpensive, less toxic and affordable trypanocides. This review discusses the current treatment of trypanosomiasis and the progress made towards the effective control of trypanosomiasis. PMID- 16316304 TI - Dopamine agonists for the treatment of restless legs syndrome. AB - The restless legs syndrome is a neurological disorder characterised by the urge to move the extremities associated with paraesthesias, which are partially or totally relieved by movement, a worsening of symptoms at rest and in the evening or at night and, as a consequence, sleep disturbances. Restless legs syndrome is common and affects 1-10% of the population. The aetiology of restless legs syndrome is unknown, but, beside genetic factors, the dopaminergic system may play a crucial role. Current treatment strategies are not curative, but may nevertheless produce an effective and lasting relief of symptoms. Evidence-based and clinical guidelines identify dopamine agonists as a first-line treatment for daily restless legs symptoms, and such substances are in the process of registration for this indication. In May 2005, the first dopamine agonist, ropinirole, was approved by the FDA. This review discusses treatment studies of dopamine agonists in patients with restless legs syndrome and focuses on long term treatment with dopamine agonists. PMID- 16316305 TI - Molecularly-targeted therapies for non-small cell lung cancer. AB - Targeting cell-signalling pathways that confer survival advantage to cancer cells has become a major focus of investigation for the treatment of various malignancies. Non-small cell lung cancer (NSCLC), a disease with wide molecular heterogeneity, has become a main testing ground for the evaluation of various targeted agents. Inhibition of the epidermal growth factor pathway with erlotinib results in improved survival and symptom control for patients with advanced NSCLC who progressed following one or two prior chemotherapy regimens. Gefitinib, the first epidermal growth factor receptor (EGFR) inhibitor to be approved by the FDA, failed to demonstrate survival advantage over placebo in a large Phase III trial for patients with advanced NSCLC. The results of this study have raised several important clinical and biological issues that may be relevant for the development of other targeted agents. Recent identification of mutations in the ATP-binding pocket of the EGFR is the first step towards proper patient selection for therapy with an EGFR tyrosine kinase inhibitor. In addition, predictive potential has also been seen with EGFR gene amplification. It is unclear whether monoclonal antibodies against the EGFR may be active independent of the EGFR mutation, as the site of action is different from tyrosine kinase inhibitors. A recent randomised clinical trial that combined the antiangiogenic agent bevacizumab with chemotherapy has demonstrated survival advantage over chemotherapy alone for certain subsets of patients with advanced NSCLC. The exciting results of this study represent an important advance in the treatment of patients with advanced NSCLC. PMID- 16316306 TI - NSAIDs and gastrointestinal complications: new clinical challenges. AB - The use of NSAIDs is associated with significant risk of upper gastrointestinal ulcer, bleeding, perforation and stricture. NSAIDs vary in their analgesic and anti-inflammatory properties; choice of NSAID, dosage and duration are the cornerstone of rationale therapy. Proton pump inhibitors are widely used to reduce the risk of serious events, despite the paucity of data that indicates that they are effective for this indication. The existing recommendations for prevention of gastrointestinal toxicity are reviewed in this article, in light of new clinical challenges posed by the emerging data regarding competing cardiovascular risk. Strategies are proposed for common clinical prescribing dilemmas, new clinical risk groups are identified and preventative strategies for these special populations are recommended. PMID- 16316307 TI - Motion sickness in migraine sufferers. AB - Motion sickness commonly occurs after exposure to actual motion, such as car or amusement park rides, or virtual motion, such as panoramic movies. Motion sickness symptoms may be disabling, significantly limiting business, travel and leisure activities. Motion sickness occurs in approximately 50% of migraine sufferers. Understanding motion sickness in migraine patients may improve understanding of the physiology of both conditions. Recent literature suggests important relationships between the trigeminal system and vestibular nuclei that may have implications for both motion sickness and migraine. Studies demonstrating an important relationship between serotonin receptors and motion sickness susceptibility in both rodents and humans suggest possible new motion sickness prevention therapies. PMID- 16316308 TI - Intravenous alteplase for acute ischaemic stroke. AB - Thrombolysis with intravenous alteplase is the only validated and approved treatment for acute ischaemic stroke. It is currently licensed for use within 3 h of stroke onset. This treatment improves functional outcome without increasing mortality, although it can initially cause a devastating intracerebral haemorrhage. Risk factors for this complication have been identified and postmarketing studies have shown an acceptable safety profile when the guidelines for drug prescription and administration are rigorously applied. Intravenous alteplase is weakly effective in recanalising major intracranial artery occlusions and more potent strategies of reperfusion are needed. Ongoing clinical trials are evaluating alteplase combined with transcranial ultrasound and intravenous microbubbles, alteplase at reduced doses combined with intravenous glycoprotein IIb/IIIa inhibitors and intravenous alteplase at a reduced dose followed by intra-arterial recanalisation. PMID- 16316309 TI - Clofarabine in the treatment of acute myeloid leukaemia and acute lymphoblastic leukaemia: a review. AB - Clofarabine, a synthesised adenosine nucleoside, has recently demonstrated single agent activity in the acute leukaemias. Originally developed to capture the best qualities of cladribine and fludarabine, clofarabine contains halogenated carbons, rendering it resistant to inactivating enzymes and maintaining its stability in acidic environments. Like other adenosine nucleosides, clofarabine acts by inhibiting ribonucleotide reductase and DNA polymerase, thereby depleting the amount of intracellular deoxynucleoside triphosphates available for DNA replication and also resulting in premature DNA chain termination. Clofarabine has also been shown to induce apoptosis in transformed cell lines, indicating that clofarabine results in cell death in both cycling and non-cycling cells. Interest in the development of clofarabine was initially hampered by the availability of other active nucleoside analogues for the treatment of haematological malignancies. However, the results of several early-phase trials evaluating the use of clofarabine in acute leukaemias in adults and children have rekindled enthusiasm for further investigation into its use. This article describes the development, pharmacology, toxicity and clinical activity of clofarabine, as well as discuss its potential role in the treatment of acute leukaemia. PMID- 16316310 TI - Docetaxel in ovarian cancer. AB - Docetaxel, a semisynthetic taxane, is a potent inhibitor of cell replication and, similar to paclitaxel, promotes in vitro assembly of stable microtubules and, therefore, prevents the depolymerisation process. Docetaxel has a higher affinity for the tubulin subunit and is associated with a 100-fold greater phosphorylation of BCL-2 inducing apoptosis. Docetaxel in combination with carboplatin demonstrates similar activity to paclitaxel/carboplatin in the upfront management of advanced ovarian cancer with less neurological, but greater haematological toxicity. This article reviews the rationale and indications for the use of docetaxel in ovarian cancer. PMID- 16316311 TI - Fluvoxamine: a selective serotonin re-uptake inhibitor for the treatment of obsessive-compulsive disorder. AB - Fluvoxamine is the selective serotonin re-uptake inhibitor with the largest database in the treatment of obsessive-compulsive disorder, a severe, and often chronic, anxiety disorder associated with substantial impairment in functioning. The selective serotonin re-uptake inhibitors represent a first-line treatment in patients with obsessive-compulsive disorder. These agents work primarily by blocking the re-uptake of serotonin into the presynaptic nerve terminal, which is believed to be mediated by their effects on the serotonin transport system. In the last two decades, the anti-obsessional effect of fluvoxamine has been tested in several double-blind, placebo-controlled and active-comparison studies, demonstrating its superior efficacy over obsessions and compulsions compared with non-serotonergic antidepressants (i.e., desipramine) and equal efficacy to clomipramine (a tricyclic antidepressant with potent serotonin re-uptake inhibition) and other selective serotonin re-uptake inhibitors (paroxetine and citalopram). However, compared with clomipramine, the selective serotonin re uptake inhibitor fluvoxamine showed fewer side effects and better tolerability. This reflects the poor affinity of this compound for adrenergic, muscarinic, cholinergic or histaminergic receptors. PMID- 16316312 TI - Levosimendan for the treatment of acute heart failure syndromes. AB - Levosimendan is a novel calcium-sensitising agent that has been shown to have beneficial inotropic, metabolic and vasodilatory effects in the treatment of acute and advanced chronic heart failure. Levosimendan binds to troponin-C in cardiomyocytes and, thereby, improves cardiac contractility without disturbing the metabolic status of the heart and increasing myocardial oxygen demand or provoking fatal cardiac arrhythmias. Levosimendan also opens ATP-sensitive potassium channels, causing peripheral arterial and venous dilatation, and increasing coronary flow reserve. When it is given as a short-term therapy, levosimendan enhances cardiac output, reduces systemic vascular resistance and lowers pulmonary capillary wedge pressure. Clinical outcomes were significantly reduced in decompensated or postmyocardial infarction heart failure patients who received levosimendan, compared with those on dobutamine or placebo. Recent investigations focusing on the anti-inflammatory and antiapoptotic actions of levosimendan in the failing heart indicate that improvement of cardiac contractile performance is closely related with the drug-induced reduction of circulating pro-inflammatory cytokines and apoptosis inducers. The most common adverse effects of levosimendan treatment are hypotension and headache. Overall, levosimendan represents an effective and safe option for the treatment of decompensated heart failure patients. PMID- 16316314 TI - Two exo-beta-D-glucosaminidases/exochitosanases from actinomycetes define a new subfamily within family 2 of glycoside hydrolases. AB - A GlcNase (exo-beta-D-glucosaminidase) was purified from culture supernatant of Amycolatopsis orientalis subsp. orientalis grown in medium with chitosan. The enzyme hydrolysed the terminal GlcN (glucosamine) residues in oligomers of GlcN with transglycosylation observed at late reaction stages. 1H-NMR spectroscopy revealed that the enzyme is a retaining glycoside hydrolase. The GlcNase also behaved as an exochitosanase against high-molecular-mass chitosan with K(m) and kcat values of 0.16 mg/ml and 2832 min(-1). On the basis of partial amino acid sequences, PCR primers were designed and used to amplify a DNA fragment which then allowed the cloning of the GlcNase gene (csxA) associated with an open reading frame of 1032 residues. The GlcNase has been classified as a member of glycoside hydrolase family 2 (GH2). Sequence alignments identified a group of CsxA-related protein sequences forming a distinct GH2 subfamily. Most of them have been annotated in databases as putative beta-mannosidases. Among these, the SAV1223 protein from Streptomyces avermitilis has been purified following gene cloning and expression in a heterologous host and shown to be a GlcNase with no detectable beta-mannosidase activity. In CsxA and all relatives, a serine aspartate doublet replaces an asparagine residue and a glutamate residue, which were strictly conserved in previously studied GH2 members with beta galactosidase, beta-glucuronidase or beta-mannosidase activity and shown to be directly involved in various steps of the catalytic mechanism. Alignments of several other GH2 members allowed the identification of yet another putative subfamily, characterized by a novel, serine-glutamate doublet at these positions. PMID- 16316315 TI - Weak acid and alkali stress regulate phosphatidylinositol bisphosphate synthesis in Saccharomyces cerevisiae. AB - Weak organic acids are used as food preservatives to inhibit the growth of spoilage yeasts, including Saccharomyces cerevisiae. Long-term adaptation to weak acids requires the increased expression of the ATP-binding cassette transporter Pdr12p, which catalyses the active efflux of the weak acids from the cytosol; however, very little is known about the signalling events immediately following application of weak acid stress. We have investigated the effects of weak acids on two stress-responsive signalling molecules, PtdIns(3,5)P2 and PtdIns(4,5)P2, which in S. cerevisiae are synthesized by Fab1p and Mss4p respectively. At low extracellular pH, benzoic acid, sorbic acid and acetic acid all cause a transient reduction in PtdIns(3,5)P2 accumulation and a more persistent rise in PtdIns(4,5)P2 levels. The increase in PtdIns(4,5)P2 levels is accompanied by a reorganization of the actin cytoskeleton. However, changes in PtdInsP2 levels are independent of weak acid-induced Pdr12p expression. In contrast, changing the extracellular medium to alkaline pH provokes a prolonged and substantial rise in PtdIns(3,5)P2 levels. As PtdIns(3,5)P2 synthesis is required for correct vacuole acidification, it is possible that levels of this molecule are modulated to maintain intracellular pH homoeostasis in response to weak acid and alkali stresses. In conclusion, we have expanded the repertoire of stress responses that affect PtdInsP2 levels to include weak acid and alkali stresses. PMID- 16316316 TI - Molecular immune mechanism of C-phycocyanin from Spirulina platensis induces apoptosis in HeLa cells in vitro. AB - C-phycocyanin (C-PC), a water-soluble protein pigment, isolated from Spirulina platensis, is of great importance because of its various medical and pharmacological properties. In the present study, we first investigated the effect of highly purified C-PC on growth and proliferation of HeLa cells in vitro. The results indicated that there was a significant decrease in the number of cells that survived for HeLa cells treated with C-PC compared with control cells untreated with C-PC. Further electron-microscopic studies revealed that C PC could induce characteristic apoptotic features, including cell shrinkage, membrane blebbing, microvilli loss, chromatin margination and condensation into dense granules or blocks. Agarose electrophoresis of genomic DNA of HeLa cells treated with C-PC showed fragmentation pattern (DNA ladder of oligomers of 180 200 bp) typical for apoptotic cells. Flow-cytometric analysis of HeLa cells treated with different concentrations of C-PC demonstrated an increasing percentage of cells in sub-G0/G1 phase. In addition, we found that C-PC could promote the expression of Fas and ICAM-1 (intercellular cell-adhesion molecule 1) protein, while it held back the Bcl-2 (B-cell lymphocytic-leukaemia proto oncogene 2) protein expression. This suggested that C-PC could induce the activation of pro-apoptotic gene and downregulation of anti-apoptotic gene expression and then facilitate the transduction of tumoural apoptosis signals that resulted in the apoptosis of HeLa cells in vitro. Caspases 2, 3, 4, 6, 8, 9, and 10 were activated in C-PC-treated HeLa cells, which suggested that C-PC induced apoptosis was caspase-dependent. C-PC treatment of HeLa cells also resulted in release of cytochrome c from the mitochondria into the cytosol that was related to apoptosis of C-PC-treated HeLa cells. PMID- 16316317 TI - Expression of adrenomedullin mRNA is altered with gestation and labour in human placenta and fetal membranes. AB - The aim of the present study was to investigate whether placental and fetal membrane AdM (adrenomedullin) mRNA expression changes with gestation and human labour, as we have previously found labour-associated changes in AdM content in fetal membranes [Al-Ghafra, Gude, Brennecke and King (2003) Clin. Sci. 105, 419 423]. Placentas and fetal membranes were collected either at term or pre-term from women either in-labour or not-in-labour, and AdM mRNA abundance was measured in tissue extracts by Northern blot analysis. Increases were found in the relative abundance of amniotic tissue AdM mRNA in both in-labour and not-in labour groups at term compared with those at pre-term, and there were positive correlations with gestational age. Relative abundance of choriodecidual tissue AdM mRNA was also significantly elevated in the not-in-labour groups between pre term and term tissues, although there was no significant correlation with gestational age. However, placental AdM mRNA expression was neither significantly increased at term (compared with pre-term) nor correlated with gestational age. In addition, there were significant increases in AdM mRNA in amnion and choriodecidua in the in-labour group compared with the not-in-labour group for both pre-term and term gestations. There was no difference in AdM mRNA in placental tissues between labour groups. In conclusion, the present study provides evidence that AdM production by fetal membranes is increased in amniotic and choriodecidual tissues at term, compared with pre-term, and in response to labour. PMID- 16316318 TI - TNF-alpha levels are associated with skin capillary recruitment in humans: a potential explanation for the relationship between TNF-alpha and insulin resistance. AB - The mechanism by which TNF-alpha (tumour necrosis factor-alpha) may cause insulin resistance is not clear. On the basis of experiments in rats, TNF-alpha has been suggested to cause defects in capillary function, with a decreased access of insulin and glucose to tissues. To test this hypothesis in humans, we assessed serum TNF-alpha concentrations, skin capillary recruitment and insulin sensitivity in a group of 37 healthy adults. In addition, we measured these variables in 21 of their prepubertal children. Serum TNF-alpha levels were measured by sandwich enzyme immunoassay, and insulin sensitivity was assessed with the hyperinsulinaemic euglycaemic clamp technique. Capillary recruitment during post-occlusive reactive hyperaemia was evaluated by videomicroscopy. In the adults, serum TNF-alpha levels were associated with both capillary recruitment (r=-0.40, P=0.02) and insulin sensitivity (r=-0.33, P=0.05). In addition, capillary recruitment was associated with insulin sensitivity (r=0.34, P=0.04). Regression analysis showed that the association between TNF-alpha and insulin sensitivity [-0.527 mg.kg(-1) of body weight.min(-1) per pmol/l per pg/ml TNF-alpha (95% confidence interval, -1.066 to 0.011); P=0.05] decreased by 30% after adjustment for capillary recruitment. In the children, neither capillary recruitment (r=0.33, P=0.2) nor insulin sensitivity (r=-0.24, P=0.4) was significantly associated with TNF-alpha. In conclusion, in adults, but not in children, serum TNF-alpha levels are associated with capillary recruitment during post-occlusive hyperaemia, which, in part, can explain the relationship between TNF-alpha and insulin resistance. Our data suggest that these relationships are initiated during growth from childhood to adulthood. PMID- 16316319 TI - Dimerization is required for SH3PX1 tyrosine phosphorylation in response to epidermal growth factor signalling and interaction with ACK2. AB - SH3PX1 [SNX9 (sorting nexin 9)] is a member of SNX super-family that is recognized by sharing a PX (phox homology) domain. We have previously shown that SH3PX1, phosphorylated by ACK2 (activated Cdc42-associated tyrosine kinase 2), regulates the degradation of EGF (epidermal growth factor) receptor. In mapping the tyrosine phosphorylation region, we found that the C-terminus of SH3PX1 is required for its tyrosine phosphorylation. Further analysis indicates that this region, known as the coiled-coil domain or the BAR (Bin-amphiphysin-Rvs homology) domain, is the dimerization domain of SH3PX1. Truncation of as little as 13 amino acid residues at the very C-terminus in the coiled-coil/BAR domain of SH3PX1 resulted in no dimerization, no ACK2-catalysed and EGF-stimulated tyrosine phosphorylation and no interaction with ACK2. The intracellular localization of SH3PX1 became dysfunctional upon truncation in the BAR domain. Taken together, our results indicate that the dimerization, which is mediated by the BAR domain, is essential for the intracellular function of SH3PX1. PMID- 16316321 TI - What we CAN do about chronic allograft nephropathy: role of immunosuppressive modulations. AB - Given the potency of modern immunosuppressive agents, kidney transplantation across alloantingen barriers is a routine phenomenon with excellent 1-year graft survival in most centers. However, the improvement in 1-year graft survival has not been matched by improvements in long-term graft function and chronic allograft nephropathy (CAN) remains the second commonest cause of graft attrition over time. Calcineurin inhibitors, namely cyclosporine A (CyA) and tacrolimus, have been implicated as causal agents in the development of the fibrotic processes that are the hallmarks of CAN. Many studies have, therefore, concentrated on the improvement of long term graft function through the modulation of immunosuppressive therapy. It is the purpose of this review to describe and appraise the available evidence for the prevention and management of CAN through modulation of immunosuppressive agents. PMID- 16316322 TI - Nuclear receptors and their coregulators in kidney. AB - Nuclear receptors are transcription factors that are essential in embryonic development, maintenance of differentiated cellular phenotypes, metabolism, and apoptosis. Dysfunction of nuclear receptor signaling leads to a wide spectra of proliferative, reproductive, and metabolic diseases, including cancers, infertility, obesity, and diabetes. In addition, many proteins have been identified as coregulators which can be recruited by DNA-binding nuclear receptors to affect transcriptional regulation. The cellular level of coregulators is crucial for nuclear receptor-mediated transcription and many coregulators have been shown to be targets for diverse intracellular signaling pathways and posttranslational modifications. This review provides a general overview of the roles and mechanism of action of nuclear receptors and their coregulators. Since progression of renal diseases is almost always associated with inflammatory processes and/or involve metabolic disorders of lipid and glucose, cell proliferation, hypertrophy, apoptosis, and hypertension, the importance of nuclear receptors and their coregulators in these contexts will be addressed. PMID- 16316323 TI - Interaction between Rf-1 and Rf-4 quantitative trait loci increases susceptibility to renal damage in double congenic rats. AB - BACKGROUND: Five quantitative trait loci (QTLs), Rf-1 to Rf-5, were found in Fawn Hooded hypertensive (FHH) rats influencing susceptibility to renal damage. Previously, we found that single transfer of the Rf-1 QTL from FHH rats onto the renal-resistant August x Copenhagen Irish (ACI) strain caused a small increase in renal susceptibility. To investigate the separate role of the Rf-4 QTL and its interaction with Rf-1, we generated a single congenic strain carrying Rf-4 and a double congenic carrying both Rf-1 and Rf-4. METHODS: Differences in renal susceptibility between ACI, Rf-1A, and Rf-4 single congenics and Rf-1A+4 double congenics were assessed using four different treatments: control (two-kidney), two-kidney with l-arginine analogue N-nitro-l-arginine methyl ester (L-NAME) induced hypertension, unilateral nephrectomy, and unilateral nephrectomy + L NAME. In separate experiments, renal blood flow (RBF) autoregulation was compared between two-kidney ACI and congenic rats. RESULTS: Compared to ACI, Rf-1A rats developed more renal damage, while Rf-4 rats did not. The most severe renal damage was found in the Rf-1A+4 double congenic rats. Analysis of variance (ANOVA) demonstrated a significant interaction between the Rf-1A and Rf-4 QTLs. The magnitude of the interaction varied with the type and duration of the treatment. The RBF autoregulation was impaired in Rf-1A single and Rf-1A+4 double congenics, while in Rf-4 single congenics it was similar to that of ACI controls. CONCLUSION: These findings indicate that the Rf-1 QTL directly influences renal susceptibility and autoregulation. In contrast, the Rf-4 QTL shows no direct effects, but significantly increases susceptibility to renal damage via an interaction with Rf-1. PMID- 16316324 TI - Glucocorticoids protect and enhance recovery of cultured murine podocytes via actin filament stabilization. AB - BACKGROUND: Nephrotic syndrome is a common kidney disease in both children and adults that is characterized by dramatic structural changes in the actin-rich foot processes of glomerular podocytes. Although glucocorticoids are the primary treatment for nephrotic syndrome, neither the target cell nor mechanism of action of glucocorticoids in nephrotic syndrome is known. For the last 30 years glucocorticoids have been presumed to act by reducing the release of soluble mediators of disease by circulating lymphocytes. In contrast, we hypothesized that glucocorticoids exert their beneficial effects in nephrotic syndrome by direct action on podocytes. METHODS: Cultured murine podocytes were treated with glucocorticoids in the presence and absence of mifepristone (to inhibit glucocorticoid-induced transcriptional activation) and challenged using our previously reported in vitro model of puromycin aminonucleoside (PAN)-induced podocyte injury, as well as by direct disruption of actin filaments with latrunculin and cytochalasin. Cell viability, actin filament distribution, total polymerized actin content, and actin-regulating guanine triphosphatase (GTPase) activities were measured. RESULTS: We demonstrated that treatment of cultured murine podocytes with the glucocorticoid dexamethasone both protected and enhanced recovery from PAN-induced injury. Dexamethasone also increased total cellular polymerized actin, stabilized actin filaments against disruption by PAN, latrunculin, or cytochalasin, and induced a significant increase in the activity of the actin-regulating GTPase RhoA. CONCLUSION: These data suggest that, contrary to the current therapeutic paradigm, the beneficial effects of glucocorticoids in nephrotic syndrome may result, at least in part, from direct effects on podocytes leading to enhanced actin filament stability. PMID- 16316325 TI - Aluminum posttranscriptional regulation of parathyroid hormone synthesis: a role for the calcium-sensing receptor. AB - BACKGROUND: Calcium regulates parathyroid hormone (PTH) gene expression by a posttranscriptional mechanism, as well as parathyroid gland growth through the activation of the calcium-sensing receptor. Aluminum decreases both parathyroid cell proliferation and PTH levels by an unknown mechanism. METHODS: To investigate the possible role of calcium-sensing receptor in the aluminum-induced PTH inhibition we used human embryonic kidney (HEK-293) cells transiently transfected with the human calcium-sensing receptor. We used a parathyroid gland tissue culture model to investigate whether the effect of aluminum in PTH mRNA was a transcriptional mechanism and also its possible role in calcium-sensing receptor expression. RESULTS: We found that Al activated the calcium-sensing receptor with higher efficiency than calcium, its biologic ligand. Aluminum inhibited PTH gene expression by a posttranscriptional mechanism, but only when low calcium is present in the medium. Finally, we found that aluminum is also able to decrease calcium-sensing receptor mRNA levels by a posttranscriptional mechanism; however, no effect was observed on calcium-sensing receptor protein. CONCLUSION: These findings indicate that aluminum impairs parathyroid function through a calcium-like mechanism due to the lack of specificity of the calcium sensing receptor. Additionally, aluminum decreases parathyroid calcium-sensing receptor mRNA levels, and the regulatory mechanism was posttranscriptional. These findings demonstrate for the first time a regulatory effect in the calcium sensing receptor by one of its ligands. PMID- 16316326 TI - Transcriptional response in the unaffected kidney after contralateral hydronephrosis or nephrectomy. AB - BACKGROUND: Unilateral loss of kidney function is followed by compensatory contralateral growth. The early, genome-wide transcriptional response of the untouched kidney to unilateral ureteral obstruction (UUO) or unilateral nephrectomy is unknown. METHODS: Twelve adult male Sprague-Dawley rats were subjected to UUO and twelve rats to unilateral nephrectomy. At time points 12, 24, and 72 hours after insult four rats each were sacrificed and the contralateral kidney harvested for genome-wide gene expression analysis, transcription factor analysis, and histomorphology. RESULTS: Microarray studies revealed that the majority of differentially expressed transcripts were suppressed in UUO and unilateral nephrectomy compared to control kidneys. The function of these suppressed genes is predominantly growth inhibition and apoptosis suggesting a net pro-hypertrophic response. Insulin-like growth factor 2 (IGF-2)-binding protein was one of the few activated genes. We observed a distinctly different molecular signature between UUO and unilateral nephrectomy at the three time points investigated. The early response in UUO rats suggests a counterbalance to the nonfiltering kidney by activation of transport pathways such as the aquaporins. Unilateral nephrectomy kidneys, on the other hand, respond immediately to contralateral nephrectomy by activation of cell cycle regulators such as the cyclin family. Several genes with weakly defined function were found to be associated with either UUO or unilateral nephrectomy. Transcription factor analysis of the identified transcripts suggests common regulation at least of some of these genes. All kidneys showed normal histology. CONCLUSION: Release of growth inhibition by nephrectomy leads to immediate cell cycle activation after unilateral nephrectomy, whereas UUO kidneys counterbalance filtration failure by activation of several transporters. PMID- 16316327 TI - Cyclosporine A and NAC on the inducible nitric oxide synthase expression and nitric oxide synthesis in rat renal artery cultured cells. AB - BACKGROUND: The immunosuppressor cyclosporine A (CsA) presents the nephrotoxicity as its major side effect that is mostly attributed to a renal vasoconstriction. This may be due to an excessive generation of vasoconstrictors like reactive oxygen species (ROS), or due to a reduction of vasodilators such as the nitric oxide, which in turn, can be caused by increased amounts of ROS. We evaluated the effect of CsA and the antioxidant N-acetylcysteine (NAC) on inducible nitric oxide synthase (iNOS) mRNA expression and nitric oxide synthesis, in rat renal artery vascular smooth muscle cells (rVSMCs) primary culture. METHODS: In cells treated during 72 hours with CsA (10 microg/mL), its vehicle (control) (10 microL/mL), Escherichia coli lipopolysaccharide (LPS) (100 microg/mL), CsA + LPS, NAC (6.13 mmol/L), or CsA + NAC, we determined the nitric oxide synthesis (Griess and chemiluminescence methods), iNOS expression [reverse transcription-polymerase chain reaction (RT-PCR)] and cell viability (acridine orange method). RESULTS: In rVSMCs, LPS increased nitric oxide and iNOS expression; CsA decreased basal and LPS-induced nitric oxide and iNOS expression; NAC increased nitric oxide and blunted the nitric oxide reduction caused by CsA, with no effect on iNOS. CsA reduced cell viability. CONCLUSION: In this study, CsA reduced nitric oxide synthesis in rVSMCs, both through iNOS down-regulation and reduction of cell viability, which could be responsible for the vasoconstrictive effect of the CsA. In the effect of CsA on nitric oxide, probably a role is also played by free radical production, as this effect was blunted by NAC. PMID- 16316328 TI - Chromosomal mapping of hyperserum IgA and glomerular IgA deposition in a high IgA (HIGA) strain of DdY mice. AB - BACKGROUND: The high IgA (HIGA) strain of ddY mice is an inbred model of IgA nephropathy (IgAN), established by selective mating of outbred ddY mice. HIGA mice show high levels of serum IgA and glomerulonephritis with mesangial IgA deposition. To identify the genetic loci responsible for hyperserum IgA and glomerular IgA deposition in this strain, quantitative trait loci analysis was carried out. METHODS: By crossing HIGA with BALB/c mice, 244 F2 generations were produced. Serum IgA levels and glomerular IgA deposition were examined at 40 weeks of age. Genetic markers were typed at 105 microsatellites and the quantitative trait loci of hyperserum IgA and glomerular IgA deposition were confirmed using Map Manager QTX software. RESULTS: Two significant quantitative trait loci of hyperserum IgA were identified on chromosome 2 [logarithm of odds (LOD) = 5.01] and chromsome 4 (LOD = 4.45), and a suggestive quantitative trait locus of hyperserum IgA was located on chromosome 1 (LOD = 3.49). On chromosome 15, a significant quantitative trait locus of glomerular IgA deposition was identified (LOD = 4.40) without the hyperserum IgA locus. Serum IgA level was weakly correlated with the intensity of glomerular IgA in 244 F2 mice; however, the quantitative trait loci of hyperserum IgA were not significantly associated with glomerular IgA deposition. CONCLUSION: These findings indicate that, in HIGA mice, glomerular IgA deposition is mainly regulated by a quantitative trait locus on chromosome 15, and hyperserum IgA synergistically but weakly affect glomerular IgA deposition. The immune disturbance similar to IgAN was revealed to be under multigenic control in HIGA mice. PMID- 16316329 TI - In high glucose protein kinase C-zeta activation is required for mesangial cell generation of reactive oxygen species. AB - BACKGROUND: We postulated that in mesangial cells exposed to high glucose, protein kinase C-zeta (PKC-zeta) is necessary for the generation of reactive oxygen species (ROS) by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and that the requirement of PKC-zeta for filamentous (F)-actin disassembly may involve ROS. To identify signaling mechanisms relevant to PKC zeta activation and ROS generation, including phosphoinositide 3 kinase (PI3 kinase), we examined mesangial cell stimulation with platelet-derived growth factor (PDGF). METHODS: In primary rat mesangial cells cultured in 5.6 mmol/L or 30 mmol/L d-glucose, PKC-zeta expression was identified with immunoblotting and activity was analyzed in cell membrane immunoprecipitates and by confocal immunofluorescence imaging. ROS generation was measured by dichlorofluorescein fluorescence using confocal microscopy and was inhibited by transfection of antisense against NADPH subunits p22(phox) or p47(phox) or with Tempol. F-actin disassembly was observed by dual-channel confocal fluorescence imaging. PI3 kinase activity was detected by immunoblotting of phosphorylated Akt. RESULTS: In high glucose, generation of NADPH oxidase-dependent ROS was dependent on PKC zeta. Conversely, sustained PKC-zeta activity was dependent on ROS generation, suggesting a positive feedback. PKC-zeta-dependent F-actin disassembly in high glucose required ROS generation. PDGF stimulated NADPH oxidase generation of ROS through a PKC-zeta mechanism that was independent of Akt phosphorylation and remained unchanged in high glucose. CONCLUSION: In high glucose, mesangial cell PKC-zeta is required for ROS generation from NADPH oxidase similar to PDGF stimulation of PKC-zeta-dependent ROS generation through a pathway independent of PI3 kinase. F-actin disassembly in high glucose also requires ROS. A positive feedback loop occurs between ROS and the activation of PKC-zeta in high glucose. PMID- 16316330 TI - Microarray and bioinformatic detection of novel and established genes expressed in experimental anti-Thy1 nephritis. AB - BACKGROUND: Microarray technology is a powerful tool that can probe the molecular pathogenesis of renal injury. In this present study microarray analysis was used to monitor serial changes in the renal transcriptome of a rat model of mesangial proliferative glomerulonephritis. Administration of anti-Thy1 antibody results in phases of acute mesangial injury (day 2), cell proliferation (day 5), matrix expansion (days 5 and 7), and subsequent healing (day 14). METHODS: Using Affymetrix (RAE230A) microarrays coupled with sequential primary biologic function-focused and secondary "baited" global cluster analysis, a cohort of established and putative novel modulators of mesangial cell turnover was identified. RESULTS: Cluster analysis of proliferative genes identified a number of gene expression profiles. The most striking pattern was increased gene expression at day 5, a cluster that included platelet-derived growth factor (PDGF), cyclins and transforming growth factor-beta (TGF-beta). The gene expression patterns identified by primary focused cluster analysis were used as bioinformatic bait and resulted in the identification of novel families of genes such as the S100 family. The expression of established and novel genes was confirmed using reverse transcription-polymerase chain reaction (RT-PCR). Next, in vivo gene expression was compared to PDGF-stimulated mesangial cells in vitro revealing similar patterns of dysregulation. CONCLUSION: Transcriptomic analysis defined both known and novel molecules involved in mesangial cell proliferation in vitro and in vivo and defined a panel of molecules that are potential contributors to mesangial cell dysfunction in glomerular disease. PMID- 16316331 TI - Kidney involvement in a nongenetic rat model of type 2 diabetes. AB - BACKGROUND: Rats fed a high fat diet and given a low dose of streptozotocin (STZ) (35 mg/kg) develop type 2 diabetes with insulin resistance, hyperinsulinemia, moderate hyperglycemia, hyperlipidemia, and salt-sensitive hypertension. We postulated that rats with noninsulinopenic (type 2) diabetes develop lesions of diabetic nephropathy significantly more prominent than those seen in classic insulinopenic (type 1) diabetic rats. METHODS: Rats were fed regular chow or high fat diet (60% calories from fat and 70% animal fat). After 5 weeks, rats fed regular chow received vehicle (controls) or 55 mg/kg STZ (type 1 diabetes mellitus). Rats fed high fat diet received vehicle (high fat) or low dose STZ, 35 mg/kg (type 2 diabetes mellitus). Rats were sacrificed 14 weeks after STZ/vehicle injection. RESULTS: Blood glucose, systolic blood pressure, and urinary protein excretion were significantly higher in both diabetes groups than in controls. Serum insulin levels (ng/mL) were higher in type 2 diabetes than in type 1 diabetes groups (0.49 +/- 0.12 vs. 0.07 +/- 0.07) (P= 0.01). Percentage of sclerosed glomeruli was significantly higher in type 2 diabetes group than in control and type 1 diabetes groups. Fibronectin expression was significantly increased in high fat, type 1 and type 2 diabetes groups compared to controls. The expression of type IV collagen, connective tissue growth factor (CTGF), and transforming growth factor-beta (TGF-beta) was significantly increased in high fat and type 2 diabetes groups compared to controls. CONCLUSION: Rats fed a high fat diet and given a low dose of STZ developed diabetes (with normal/high insulin levels), hypertension, and proteinuria. Kidney lesions in this type 2 model appear to be more pronounced than in type 1 diabetic rats despite lower blood glucose levels and proteinuria. We present a nongenetic rat model of type 2 diabetes mellitus and nephropathy. PMID- 16316332 TI - Determinants of tubular bone marrow-derived cell engraftment after renal ischemia/reperfusion in rats. AB - BACKGROUND: Ischemia/reperfusion (I/R) injury is a major cause of acute renal failure (ARF). ARF is reversible, due to an innate regenerative process, which is thought to depend partly on bone marrow-derived progenitor cells. The significance of these cells in the repair process has been questioned in view of their relatively low frequency. Here, we hypothesize that the severity of renal damage and the postischemic recovery time are determinants of tubular bone marrow derived cell (BMDC) engraftment. METHODS: We used a model of unilateral renal I/R in F344 rats reconstituted with R26-human placental alkaline phosphatase (hPAP) transgenic bone marrow, in which we quantified and characterized tubular BMDC engraftment with increasing severity of damage and in time. RESULTS: After I/R injury, BMDC engrafted the tubular epithelium and acquired an epithelial phenotype. Tubular epithelial BMDC engraftment increased with longer ischemic time, indicating that tubular epithelial BMDC engraftment increases with the severity of damage. The number of circulating progenitor cells doubled early after I/R injury and was followed by a transient increase in tubular epithelial BMDC engraftment. The latter positively correlated with morphological recovery of the kidney over time. CONCLUSION. The extent of tubular BMDC engraftment depends on the severity of renal damage and follows a distinct time course after I/R injury. Therefore, the severity of damage and time course need to be taken into account when interpreting data on the role of tubular BMDC engraftment in renal repair after I/R injury. PMID- 16316333 TI - Toll-like receptor-4: renal cells and bone marrow cells signal for neutrophil recruitment during pyelonephritis. AB - BACKGROUND: The molecular mechanisms of pathogen recognition that initiate infective pyelonephritis are poorly understood. Toll-like receptor-4 (TLR4) mutant mice infected with uropathogenic Escherichia coli lack renal CXCL2 mRNA expression, subsequent neutrophil recruitment, and renal abscess formation. METHODS: We used a bone marrow transplant approach in order to investigate the contribution of TLR4 in intrinsic renal cells or bone-marrow-derived immune cells to neutrophil recruitment during infective pyelonephritis. RESULTS: Both chimera either expressing mutant tlr4 in intrinsic renal cells and wild-type tlr4 in bone marrow-derived cells or vice versa showed an impaired response to uropathogenic E. coli infection in terms of leukocyturia and renal abscess formation when compared to tlr4 wild-type mice with congenic bone marrow transplants. CONCLUSION: These data suggest that TLR4 is required on both intrinsic renal cells (e.g., tubular epithelial cells) and bone marrow-derived immune cells for the control of ascending uropathogenic E. coli infection by initiating chemokine driven renal neutrophil recruitment. PMID- 16316334 TI - Prediction of urinary protein markers in lupus nephritis. AB - BACKGROUND: Lupus nephritis is divided into six classes and scored according to activity and chronicity indices based on histologic findings. Treatment differs based on the pathologic findings. Renal biopsy is currently the only way to accurately predict class and activity and chronicity indices. We propose to use patterns of abundance of urine proteins to identify class and disease indices. METHODS: Urine was collected from 20 consecutive patients immediately prior to biopsy for evaluation of lupus nephritis. The International Society of Nephrology/Renal Pathology Society (ISN/RPS) class of lupus nephritis, activity, and chronicity indices were determined by a renal pathologist. Proteins were separated by two-dimensional gel electrophoresis. Artificial neural networks were trained on normalized spot abundance values. RESULTS: Biopsy specimens were classified in the database according to ISN/RPS class, activity, and chronicity. Nine samples had characteristics of more than one class present. Receiver operating characteristic (ROC) curves of the trained networks demonstrated areas under the curve ranging from 0.85 to 0.95. The sensitivity and specificity for the ISN/RPS classes were class II 100%, 100%; III 86%, 100%; IV 100%, 92%; and V 92%, 50%. Activity and chronicity indices had r values of 0.77 and 0.87, respectively. A list of spots was obtained that provided diagnostic sensitivity to the analysis. CONCLUSION: We have identified a list of protein spots that can be used to develop a clinical assay to predict ISN/RPS class and chronicity for patients with lupus nephritis. An assay based on antibodies against these spots could eliminate the need for renal biopsy, allow frequent evaluation of disease status, and begin specific therapy for patients with lupus nephritis. PMID- 16316335 TI - Biochemical monitoring of mTOR inhibitor-based immunosuppression following kidney transplantation: a novel approach for tailored immunosuppressive therapy. AB - BACKGROUND: Immunosuppressive therapy with the mammalian target of rapamycin (mTOR) inhibitors requires a fine balance between allograft maintenance and drug related side effects. METHODS: In this study we examined the feasibility of monitoring TOR inhibitor-based immunosuppression by assessment of the phosphorylation status at the Thr(389) site of the p70S6 kinase in peripheral blood mononuclear cells (PBMCs). At total of 36 patients with renal transplants and 8 healthy controls were enrolled. RESULTS: We found that sirolimus treatment was associated with a pronounced inhibition of p70S6 kinase phosphorylation, as compared to healthy donors or otherwise immunosuppressed patients. In sirolimus treated patients, phosphorylation of the p70S6 kinase was significantly inhibited when sirolimus trough levels were > 6 ng/mL. In contrast, for trough levels <6 ng/mL, the degree of inhibition of p70S6 kinase phosphorylation showed a high degree of interindividual variability. We recorded a total of five clinical relevant rejection episodes in this patient category. Intriguingly, rejecters uniformly maintained a high degree of phosphorylation independent of the sirolimus trough level whereas non-rejecters showed a significant inhibition of phosphorylation. CONCLUSION: Therefore, the phosphorylation status of the p70S6 kinase appears to provide more relevant information on the desired effect of sirolimus in target cells as compared to trough level measurements. Moreover, this assay provides an opportunity to safely titer down sirolimus levels to avoid overimmunosuppression and, on the other hand, to identify patients with insufficient TOR inhibitor therapy that are at risk for rejection. PMID- 16316336 TI - Rapamycin worsens renal function and intratubular cast formation in protein overload nephropathy. AB - BACKGROUND: Rapamycin (sirolimus) is associated with functional nephrotoxicity in some patients with nephrotic glomerular diseases but the pathophysiologic mechanisms are not known. This study investigated the effects of rapamycin on renal function and structure in protein overload nephropathy. METHODS: Rats with protein overload nephropathy [induced by bovine serum albumin (BSA), 2.1 g by daily intraperitoneal injection, day 0 to day 3] received daily intraperitoneal injections of either vehicle [dimethyl sulfoxide (DMSO)], rapamycin (0.2 mg/kg, an inhibitor of mammalian target of rapamycin), or roscovitine (3.5 mg/kg, a small molecule cyclin-dependent kinase inhibitor) (N= 9 each) from day -3 to day 3. RESULTS: In protein overload nephropathy, rapamycin caused severe acute renal failure and mild hypercholesterolemia (both P < 0.05). Rapamycin dramatically increased intratubular cast formation, and proximal tubular epithelial cells were swollen and engorged with increased cytoplasmic protein droplets. The number of 5 bromo-2'-deoxyuridine (BrdU)-positive tubular epithelial cells increased by more than 20-fold on day 3 in protein overload nephropathy, and this was attenuated by 65% with rapamycin (P < 0.05), whereas roscovitine was ineffective. Rapamycin increased the protein expression of p27(kip1) in tubular epithelial cells, but did not alter D-type cyclin expression or apoptosis. CONCLUSION: Rapamycin caused a specific pattern of acute renal injury characterized by increased intratubular cast formation in protein overload nephropathy. This could be due to disruption of a potentially important compensatory mechanism in nephrotic glomerular diseases involving tubular epithelial cell protein endocytosis and proliferation. PMID- 16316337 TI - Role of altered renal lipid metabolism and the sterol regulatory element binding proteins in the pathogenesis of age-related renal disease. AB - BACKGROUND: There are well-known changes in age-related renal function and structure, including glomerulosclerosis and decline in glomerular filtration rate (GFR). The purpose of this study was to identify a potential role for lipids in mediating age-related renal disease. METHODS: Mice of five different age groups (3, 6, 12, 19, and 23 months old) were studied. RESULTS: We have found that in C57BL/6 mice there was a progressive increase in age-related glomerulosclerosis [increase in periodic acid-Schiff (PAS) staining and accumulation of extracellular matrix proteins including type IV collagen and fibronectin], increased glomerular basement thickness and podocyte width and effacement, and increased proteinuria. These changes were associated with age-related increase in lipid accumulation as determined by increased Oil Red O staining in kidney sections. Biochemical analysis indicated that these lipid deposits corresponded to significant increases in renal triglyceride and cholesterol content. We have also found significant age-related increases in the nuclear transcription factors, sterol regulatory element-binding proteins (SREBP-1 and SREBP-2), protein abundance and increased expression or activity of their target enzymes that play an important role in lipid synthesis. CONCLUSION: Our results indicated that there was an age-related increase in renal expression of SREBP-1 and SREBP-2 with resultant increases in lipid synthesis and triglyceride and cholesterol accumulation in the kidney. Because we have previously shown that increased expression of SREBPs in the kidney per se results in glomerulosclerosis and proteinuria, our data suggested that increased SREBPs' expression resulting in increased renal lipid accumulation may play an important role in age-related nephropathy. PMID- 16316338 TI - Immunolocalization of fibroblast growth factor-1 (FGF-1), its receptor (FGFR-1), and fibroblast-specific protein-1 (FSP-1) in inflammatory renal disease. AB - BACKGROUND: The fibroblast growth factor (FGF) family has functions in development, cell proliferation, migration, and differentiation. While FGF-2 induces fibrosis, the role of FGF-1 in inflammation and fibrosis is less defined. We examined the expression of FGF-1 and FGF receptor (FGFR-1) to determine if renal diseases with varying etiologies of inflammation, including lupus nephritis (LN), acute interstitial nephritis (AIN) and acute rejection superimposed on chronic allograft nephropathy (CAN), showed varying patterns of expression. We also examined the expression of fibroblast-specific protein-1 (FSP-1), which has been linked to epithelial-mesenchymal transition (EMT) and fibrosis, to determine whether it was linked to potential profibrotic and inflammatory FGF-1 mechanisms. METHODS: Proliferative LN (PLN) (N= 12), nonproliferative lupus nephritis (NPLN) (N= 5), AIN (N= 6), CAN (N= 4), and normal kidneys (N= 3) were studied. FGF, FGFR 1, and FSP-1 were localized by immunohistochemistry, and intensity scored on a 0 to 3+ scale. Double staining with CD68 and separate immunohistochemical staining for CD4 and CD8 with serial sections analysis were done to identify if T lymphocytes or macrophages showed staining for FGF-1 and FGFR-1 or FSP-1. RESULTS: In normal kidneys, FGF-1 was expressed in mesangial cells (0.67 +/- 0.58), glomerular endothelial (0.67 +/- 0.58), visceral, and parietal epithelial cells (1.67 +/- 0.58). FGFR-1 showed a similar pattern of staining but also was expressed in tubular epithelium, and arterial endothelium and smooth muscle. Expression of FGF-1 was increased over normal in glomerular parenchymal cells only in CAN in podocytes (2.30 +/- 0.58 vs. 3.00 +/- 0.00) (P < 0.05) and parietal epithelial cells (1.67 +/- 0.58 vs. 2.25 +/- 0.50) (P < 0.05). Infiltrating glomerular and interstitial inflammatory cells in diseased glomeruli also expressed FGF-1 and FGFR-1. Tubular cells expressed slightly increased FGFR 1 in renal diseases vs. normal, whereas tubules remained negative for FGF-1 in diseased kidneys. FSP-1 expression was prominent in the interstitium in all kidneys with interstitial inflammation, and most prominent in CAN. Interstitial FSP-1+ cells were consistent with a myofibroblast-type morphology, and did not stain with CD-68. FSP-1 expression was closely associated with inflammatory cells expressing FGF-1 and FGFR-1. FSP-1 also showed positivity within crescents and occasional podocytes in PLN. CONCLUSION: The expression of FGF-1 and FGFR-1 in infiltrating lymphocytes and macrophages, and of FGFR-1 in tubules, is supportive, but does not prove causality, of the possibility that FGF-1 might have both autocrine and paracrine functions in renal inflammation. However, the initial stimulus for renal inflammation, whether immune complex, hypersensitivity or rejection, did not alter expression patterns of FGF-1 or its receptor. The colocalization of inflammatory infiltrates with interstitial fibrosis supports the possibility of a contribution of FGF-1 for chemotaxis and associated fibrosis, further supported by interstitial FSP-1 expression closely associated with these inflammatory cells expressing FGF-1 and FGFR-1. PMID- 16316339 TI - Lipid peroxidation is not the underlying cause of renal injury in hyperoxaluric rats. AB - BACKGROUND: Hyperoxaluria is a major risk factor of calcium oxalate stone disease and renal injury is thought to be a significant initiating event. However, the relationship among oxidative stress, renal tubule injury and hyperoxaluria in the progression of nephrolithiasis is unclear, especially in animal models. In the current study, we assess the role of oxidative stress in renal tubular damage in a rat model of chronic hyperoxaluria (HYP) and chronic renal failure induced by hyperoxaluria (HRF) compared to control rats. METHODS: Urinary excretion of renal tubular enzymes, including lactate dehydrogenase (LDH), alkaline phosphatase (AP), N-acetyl-beta-D-glucosaminidase (NAG), and alpha- and mu-glutathione-S transferase (alpha-GST and mu-GST, respectively) was quantified in four groups of Sprague-Dawley rats. The study included normal controls, those made hyperoxaluric with ethylene glycol administration (HYP), unilateral nephrectomized controls, and unilateral nephrectomized rats administered ethylene glycol (HRF). Levels of catalase, superoxide dismutase (SOD), glutathione peroxidase (GP), and glutathione transferase (GST) in the renal cortex were measured after 4 weeks and lipid peroxidation was assessed by measuring 8-isoprostane in the urine and lipid hydroperoxide in the renal cortex. RESULTS: Urinary excretion of NAG, AP, and LDH was elevated after 2 and 4 weeks in the HYP and HRF groups. Urinary levels of mu GST, a marker of distal tubule damage, were elevated in HRF rats after 4 weeks. alpha-GST levels were similar between control and HYP rats but were lower in HRF rats. Levels of catalase, SOD, GP, and GST in the renal cortex were similar among control, HYP, and unilateral nephrectomized control rats, but were attenuated in the HRF rats after 4 weeks. Renal cortical content of lipid hydroperoxide and urinary 8-isoprostane levels were similar among all groups after 4 weeks. CONCLUSION: Ethylene glycol-induced hyperoxaluria in Sprague-Dawley rats is accompanied by enzymuria, which is suggestive of renal tubular damage. The antioxidant capacity of the renal cortex in HYP rats is similar to that of control rats after 4 weeks of treatment; however, this capacity is significantly attenuated in rats that are in renal failure induced by hyperoxaluria, although significant lipid peroxidation is not evident. These results suggest that lipid peroxidation is not the underlying cause of renal injury in hyperoxaluric rats. PMID- 16316340 TI - Involvement of endoplasmic reticulum (ER) stress in podocyte injury induced by excessive protein accumulation. AB - BACKGROUND: An imbalance between protein load and folding capacity is referred to as endoplasmic reticulum (ER) stress. As a defense mechanism, cells express ER stress inducible chaperons, such as oxygen-regulated proteins 150 (ORP150) and glucose-regulated proteins (GRPs). While ER stress is important in various diseases, a pathophysiologic role for ER stress in kidney disease remains elusive. Here we investigate expression of ER stress proteins in cultured rat podocytes as well as in our recently developed animal model of abnormal protein retention within the ER of podocytes (i.e., megsin transgenic rat). METHODS: The expression of ER stress inducible proteins (ORP150, GRP78, or GRP94) in cultured podocytes treated with tunicamycin, A23187, SNAP, hypoxia, or hyperglycemia, and the renal tissues or isolated glomeruli from megsin transgenic rats was analyzed by Western blotting analysis, immunohistochemistry, or confocal microscopy. RESULTS: Cultured podocytes demonstrated that treatment with tunicamycin, A23187, and SNAP, but not hypoxia or hyperglycemia, up-regulate expression of ER stress proteins. Extracts of isolated glomeruli from megsin transgenic rats reveal marked up-regulation of ER stress chaperones in podocytes, which was supported by immunohistochemical analysis. Confocal microscopy revealed that ER stress in podocytes was associated with cellular injury. Podocytes of transgenic rats overexpressing a mutant megsin, without the capacity for polymerization within the ER, do not exhibit ER stress or podocyte damage, suggesting a pathogenic role of ER retention of polymerized megsin. CONCLUSION: This paper implicates a crucial role for the accumulation of excessive proteins in the podocyte ER in the induction of ER stress and associated podocyte injury. PMID- 16316341 TI - Plasminogen activator inhibitor-1 gene deficiency attenuates TGF-beta1-induced kidney disease. AB - BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) stimulates the deposition of extracellular matrix (ECM), which is a hallmark in end-stage renal disease. The importance of TGF-beta1-induced changes in protease activity in this process is not fully elucidated. TGF-beta1 up-regulates plasminogen activator inhibitor type 1 (PAI-1), which lowers matrix degradation. Our aim was to investigate the importance of PAI-1 in TGF-beta1-induced kidney disease. METHODS: TGF-beta1 transgenic mice were bred with PAI-1 gene deficient mice. The effect of PAI-1 gene knockout on TGF-beta1-induced glomerular disease was investigated by measuring morphologic changes in the glomeruli. Interstitial changes were assessed by measurement of total collagen content and expression and localization of ECM components. Finally, protease activity was evaluated by plasmin activity measurement and by gel and in situ gelatin zymography. RESULTS: TGF-beta1 elevated PAI-1 expression fourfold. PAI-1 gene deficiency attenuated the TGF beta1-induced mesangial expansion and basement membrane thickening. Furthermore, PAI-1 knockout diminished collagen accumulation in TGF-beta1-positive mice. The expression of both collagen type I and III were reduced. Interestingly, no difference in protease activity could be ascertained as cause of the decreased ECM accumulation. CONCLUSION: We show that PAI-1 gene deficiency attenuates TGF beta1-induced kidney disease, decreasing both glomerular and interstitial ECM deposition. Thus, PAI-1 mediates some of the biological effects of TGF-beta1 in vivo. However, we could not find evidence supporting the notion that the effect was mediated through increased protease activity. PMID- 16316342 TI - Transcriptional analysis of the molecular basis of human kidney aging using cDNA microarray profiling. AB - BACKGROUND: The molecular basis of renal aging is not completely understood. METHODS: We used global gene expression monitoring by cDNA microarrays to identify age associated genes in human kidney samples. Our samples included young (8 weeks-8 years, N= 4), adult (31-46 years, N= 7), and old kidneys (71-88 years, N= 9). RESULTS: Old kidneys had more glomerulosclerosis, tubular atrophy, interstitial fibrosis, and fibrous intimal thickening in small arteries. We identified approximately 500 genes that were differentially expressed among the three age groups. Old kidneys appeared to have increased extracellular matrix turnover and a nonspecific inflammatory response, combined with a reduction in processes dependent on energy metabolism and mitochondrial function. Quantitative supervised bioinformatics analyses of adult and old kidney expression data correlated the expression of 255 gene profiles with renal pathology scores. Microarray class prediction analysis (PAM) identified 50 unique genes that segregated old kidneys into two distinct clusters: those more similar within age class (OO, N= 5) versus old kidneys more similar to adult kidneys (OA, N= 4). The expression of six functionally significant genes was further validated by quantitative reverse transcription-polymerase chain reaction (RT-PCR) (FN1, MMP7, TNC, SERPIN3A, BPHL, CSPG2) in the experiment group and, subsequently, confirmed independently in 17 additional old and adult age-stratified test kidney samples. The p53 inducible gene, CSPG2, performed best in separating OO kidneys from adults and OA samples in this analysis. CONCLUSION: The method described in this study using independent validation samples can be envisioned to test utility of the identified genes in assessing age-related changes that contribute to decline in renal function. PMID- 16316343 TI - Fibrate prevents cisplatin-induced proximal tubule cell death. AB - BACKGROUND: In previous studies we have shown that cisplatin inhibits peroxisome proliferator-activated receptor-alpha (PPAR-alpha) activity and consequently fatty acid oxidation, and these events precede proximal tubule cell death. In addition the use of fibrate class of PPAR-alpha ligands ameliorate renal function by preventing both inhibition of fatty acid oxidation and proximal tubule cell death. METHODS: LLC-PK1 cells were treated with cisplatin and apoptosis was established by the presence of nuclear fragmentation and by cell cycle analysis. Proximal tubular cells treated with cisplatin and bezafibrate were subjected to sub cellular fractionation and the presence of Bax, Bcl-2, cytochrome c, and active caspase-3 in the cytosolic and mitochondrial membrane fractions was determined by Western blot analysis. PPAR-alpha activity was measured by determining luciferase activity after transfection of LLC-PK1 cells with TK-Luc 3x PPAR response elements (PPRE), and the accumulation of nonesterified free fatty acids was measured in lysates obtained from cells treated with cisplatin and bezafibrate. RESULTS: Incubation of LLC-PK1 cells with 25 micromol/L cisplatin for 18 hours induced 41.5% apoptosis measured by cell cycle analysis. Cisplatin-induced apoptosis was significantly suppressed by bezafibrate, a fibrate class of PPAR-alpha ligand. Bezafibrate treatment of LLC-PK1 cells prevented cisplatin-induced translocation of proapoptotic Bax from the cytosol to the mitochondrial fraction, and increased the expression of antiapoptotic molecule Bcl-2. Cisplatin-induced inhibition of PPAR-alpha activity was accompanied by increased accumulation of nonesterified free fatty acids. Pretreatment with bezafibrate prevented both the inhibition of PPAR-alpha activity and the accumulation of nonesterified free fatty acids induced by cisplatin. Finally, bezafibrate prevented cisplatin-induced release of cytochrome c from the mitochondria to the cytosol, and the cleavage of procaspase-3 to active caspase-3. CONCLUSION: Bezafibrate treatment inhibits cisplatin-mediated tubular injury by preventing the activation of various cellular mechanisms that lead to proximal tubule cell death. These findings support our previous observations where the use of fibrates represents a novel strategy to ameliorate proximal tubule cell death in cisplatin-induced acute renal failure. PMID- 16316344 TI - Calcium-binding proteins annexin A2 and S100A6 are sensors of tubular injury and recovery in acute renal failure. AB - BACKGROUND: Rise in cellular calcium is associated with acute tubular necrosis, the most common cause of acute renal failure (ARF). The mechanisms that calcium signaling induce in the quiescent tubular cells to proliferate and differentiate during acute tubular necrosis have not been elucidated. METHODS: Acute tubular necrosis induced in mice by single intravenous injection of uranyl nitrate and examined after 1, 3, 7, and 14 days. Renal function was monitored and kidneys were evaluated by histology, immunohistochemistry, Western blotting, in situ hybridization, and real-time reverse transcription-polymerase chain reaction (RT PCR). Models of folic acid induced-ARF and ischemic/reperfusion (I/R) injury were similarly investigated. RESULTS: Analysis of mRNA expression of intracellular calcium and phospholipid-binding proteins demonstrated selective expression of S100A6 and Annexin A2 (Anxa2) in the renal cortex with marked elevation on day 3, and gradually decline on day 7 and further attenuation on day 14. Similarly, the expression of both proteins, as demonstrated by immunohistochemistry and Western blot analysis, was increased and reached the peak level on day 7 and then gradually declined by day 14. Vimentin, a marker of dedifferentiated cells, was highly expressed during the recovery phase. Combined in situ hybridization immunohistochemistry revealed colocalization of both S100A6 and Anxa2 with proliferating cell nuclear antigen (PCNA). The universality of this phenomenon was confirmed in two other mouse acute tubular necrosis models, the ischemic reperfusion injury and folic acid-induced ARF. CONCLUSION: Collectively, these findings demonstrate that S100A6 and Anxa2 expression, initiated in response to tubular injury, persist in parallel throughout the recovery process of tubular cells in acute renal failure. PMID- 16316345 TI - Altered expression of rat renal cortical OAT1 and OAT3 in response to bilateral ureteral obstruction. AB - BACKGROUND: Bilateral ureteral obstruction (BUO) is characterized by the development of hemodynamic and tubular lesions. However, little is known about the expression of organic anion renal transporters. The objective of this work was to study the renal excretion of p-aminohippurate (PAH) and the cortical renal expression of the organic anion transporter 1 (OAT1) and organic anion transporter 3 (OAT3) in BUO rats. METHODS: Male Wistar rats underwent bilateral obstruction of the proximal ureters for 24 hours (BUO) or sham operation. After 24 hours of ureteral releasing, the following studies were performed: PAH renal excretion employing conventional clearance techniques and OAT1 and OAT3 abundance (homogenates, intracellular and basolateral plasma membrane fractions from renal cortex) using immunoblotting and immunocytochemical techniques (light microscopic and confocal immunofluorescence microscopic analysis). RESULTS: BUO rats showed a lower renal excretion of PAH. In obstructed kidneys, immunoblotting revealed a significant decrease in the abundance of both OAT1 and OAT3 in basolateral plasma membranes from renal cortex. An increase of OAT1 expression was observed in homogenates and in intracellular membrane fractions in kidneys from BUO rats compared with sham-operated ones, indicating an internalization of this carrier. Immunocytochemical techniques confirmed these results. On the contrary, OAT3 expression was reduced both in homogenates and in intracellular membrane fractions in obstructed kidneys. CONCLUSION: BUO was associated with down regulation of OAT1 and OAT3 in basolateral plasma membranes from proximal tubule cells, thus these carriers may play important roles in the impaired organic anion excretion displayed in the obstructed kidney. PMID- 16316346 TI - Induction of protective genes by cobalt ameliorates tubulointerstitial injury in the progressive Thy1 nephritis. AB - BACKGROUND: We previously demonstrated that chronic hypoxia has pivotal roles in the progression of tubulointerstitial injury from the early stage of the uninephrectomized Thy1 nephritis model. We have also shown that pretreatment of cobalt confers renoprotection in the ischemia/reperfusion (I/R) injury, in association with the up-regulation of hypoxia-inducible factor (HIF)-regulated genes. Here, we tested the hypothesis that cobalt administration not only attenuates acute ischemic insult, but also ameliorates tubulointerstitial injury secondary to chronic hypoxia. METHODS: We applied sustained cobalt treatment to the uninephrectomized Thy1 nephritis model at 3 to 5 weeks, when tubular hypoxia appeared. Histologic evaluation, including glomerular and peritubular capillary networks, was made at 8 weeks. HIF activation was confirmed by real-time polymerase chain reaction (PCR) analyses for HIF-regulated genes, such as erythropoietin (EPO), vascular endothelial growth factor (VEGF), and heme oxygenase 1 (HO-1). Up-regulation of HIF-1alpha and HIF-regulated genes was also verified by Western blotting analysis. To elucidate responsible mechanisms of cobalt in the amelioration of tubuloniterstitial injury, terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) staining was conducted at 5 weeks. A combination therapy with angiotensin receptor blocker (ARB), olmesartan, was also challenged. RESULTS: Although the intervention did not change glomerular structural damage or urinary protein excretion rate, tubulointerstitial injury was improved in cobalt treated animals when compared with the vehicle-treated group. The amelioration was associated with the parallel up-regulation of renoprotective, HIF-regulated gene expression. TUNEL staining revealed that the number of apoptotic cells was reduced in the cortex by cobalt administration, suggesting that renoprotection was achieved partly through its antiapoptotic properties. Furthermore, it was demonstrated that cobalt treatment exerts additional renoprotective effects with the ARB treatment in this model. CONCLUSION: Maneuvers to activate HIF in the ischemic tubulointerstitium will be a new direction to future therapeutic strategies. PMID- 16316347 TI - Constriction velocities of renal afferent and efferent arterioles of mice are not related to SMB expression. AB - BACKGROUND: Constriction of renal arterioles contributes significantly to the control of perfusion and glomerular filtration. Afferent but not efferent arterioles express smooth muscle myosin heavy chain B (SMB) (with a 5'-insert of seven amino acids). The aim of the present study was to investigate (1) the constriction characteristics of afferent and efferent arterioles under physiologic load and (2) whether expression of SMB may causally contribute to these constriction characteristics. METHODS: We compared constriction parameters [constriction amplitude, maximal rate of constriction velocity ("dc/dt(max)"), and time to half-maximal constriction (t(1/2)) of in vitro perfused renal afferent and efferent arterioles of wild-type (smb(+/+)] and homozygous SMB knockout [smb(-/-)] mice upon stimulation with angiotensin II (Ang II) (10(-8) mol/L) and potassium chloride (KCl) (100 mmol/L). SMB expression was investigated by double-labeling immunofluorescence. RESULTS: Contraction amplitude and dc/dt(max) of mouse afferent arterioles upon Ang II stimulation were significantly greater compared to efferent arterioles. However, constriction amplitudes, dc/dt(max), and t(1/2) of afferent as well as efferent arterioles upon Ang II stimulation were similar in smb(+/+) and smb(-/-) mice. Constriction amplitudes upon KCl stimulation of afferent arterioles were similar in both smb(+/+) and smb(-/-) mice. Furthermore, KCl-induced dc/dt(max) and t(1/2) of afferent arterioles were similar in both smb(+/+) and smb(-/-) mice. SMB expression could be detected in afferent but not efferent arterioles in smb(+/+) mice. No SMB expression in either arteriole could be observed in smb(-/-) mice. CONCLUSION: Our results suggest that the presence of different alternatively 5' spliced smooth muscle-myosin heavy chain (SM-MHC) isoforms does not dominate the different contractile features of physiologically loaded renal afferent or efferent arterioles. PMID- 16316348 TI - Nitric oxide preserves the glomerular protein permeability barrier by antagonizing superoxide. AB - BACKGROUND: The interaction of nitric oxide with superoxide (O2-) is a major O2- scavenging mechanism that can minimize O2 (-)-mediated oxidative stress. Glomeruli produce both nitric oxide and O2- and generation of both radicals is increased in various forms of glomerular disease. O2- increases glomerular capillary permeability to albumin (P(alb)). The present studies tested the hypothesis that nitric oxide opposes this effect, thereby preserving the glomerular protein permeability barrier. METHODS: P(alb) was determined in isolated rat glomeruli by measuring the change in glomerular volume in response to an experimental oncotic gradient. Changes in P(alb) in response to O2- generated by tumor necrosis factor-alpha (TNF-alpha) or xanthine/xanthine oxidase (X/XO) was assessed under conditions of nitric oxide depletion and repletion. RESULTS: Incubation of rat glomeruli with the nitric oxide synthase (NOS) inhibitor L-N(G)-monomethyl arginine (L-NMMA) increased P(alb.) This effect was reversed by the nitric oxide donor diethylenetriamine NONOate (DETA-NONOate) and by the superoxide dismutase (SOD) mimetic Tempol. O2- generated after incubation with TNF-alpha or X/XO increased P(alb). This effect was blocked by DETA-NONOate. CONCLUSION: We demonstrate that nitric oxide protects the glomerular filtration barrier from injury caused by O2- and suggest that inhibition of nitric oxide synthesis could enhance O2(-)-mediated oxidative injury under pathologic conditions. PMID- 16316349 TI - Increase in heme oxygenase-1 levels ameliorates renovascular hypertension. AB - BACKGROUND: The heme oxygenase system (HO-1 and HO-2) catalyzes the conversion of heme to free iron, carbon monoxide (CO), a vasodepressor, and biliverdin, which is further converted to bilirubin, an antioxidant. HO-1 induction has been shown to suppress arachidonic acid metabolism by cytochrome P450 (CYP450) monooxygenases and cyclooxygenases (COX), and to decrease blood pressure in spontaneously hypertensive rats (SHR). The Goldblatt 2K1C model is a model of renovascular hypertension in which there is increased expression of COX-2 in the macula densa and increased renin release from the juxtaglomerular apparatus of the clipped kidney. We examined whether HO-1 overexpression, as a prophylactic approach, would attenuate renovascular hypertension and evaluated potential mechanisms that may account for its effect. METHODS: 2K1C rats were treated with cobalt protoporphyrin (CoPP) or tin mesoporphyrin (SnMP) one day before surgery and weekly for three weeks thereafter. We measured systolic blood pressure, HO activity, HO-1, HO-2, COX-1 and COX-2 protein expression, heme content, and nitrotyrosine levels as indices of oxidative stress. Urinary prostaglandin excretion (PGE2), plasma renin activity (PRA), and plasma aldosterone levels were also measured. RESULTS: CoPP administration induced renal HO-1 expression by 20 fold and HO activity by 6-fold. This was associated with a reduction in heme content, nitrotyrosine levels, COX-2 expression and urinary PGE2 excretion, and attenuation of the development of hypertension in the 2K1C rats. There was no decrease in plasma renin activity; however, plasma aldosterone levels were significantly lower. In the 2K1C SnMP-treated rats, blood pressure was significantly higher than that of untreated 2K1C rats throughout the study, and the difference in the size of the smaller left clipped kidney compared to the nonclipped right kidney was significantly increased. CONCLUSION: These findings define an action of prolonged HO-1 induction to interrupt and counteract the influence of the renin-angiotensin-aldosterone system (RAAS) to increase in blood pressure in the 2K1C model of renovascular hypertension. Multiple mechanisms include a decrease in oxidative stress as indicated by the decrease in cellular heme and nitrotyrosine levels, an anti-inflammatory action as evidenced by a decrease in COX-2 and PGE2, interference with the action of angiontensin II (Ang II) as evidenced by an increase in PRA in the face of a decrease in PGE2 and aldosterone, as well as the inhibition of aldosterone synthesis. PMID- 16316350 TI - Glycated and carboxy-methylated proteins do not directly activate human vascular smooth muscle cells. AB - BACKGROUND: Advanced glycation end products (AGEs) accumulate in patients with diabetes, particularly at sites of vascular damage and within atherosclerotic lesions, but whether they have direct actions on vascular smooth muscle cells (VSMCs) is controversial. METHODS: AGEs were constructed and characterized by protein content, level of modification, fluorescence, and molecular size. Human VSMCs were derived from different vascular beds. Glucose consumption, de novo protein synthesis, and proteoglycan biosynthesis were measured using a colorimetric assay and metabolic radiolabeling. Receptor for AGEs (RAGE) expression was assessed by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: Treatment with AGEs under low or high glucose conditions showed no change in cellular glucose consumption or in cellular protein synthesis under low glucose conditions. Treatment of VSMCs with Nepsilon-(carboxymethyl)lysine in the presence of low glucose increased [35S] sulfate incorporation into secreted proteoglycans by 72% (P < 0.001) and 67% (P < 0.001); however, the control proteins also increased [35S]-sulfate incorporation into proteoglycans by 56% (P < 0.01), with similar effects observed under high glucose conditions. Human VSMCs showed no difference in response to glycated and non-glycated protein. Protein and gene expression of RAGE in VSMC was approximately 50-fold lower compared to HMEC-1 and U937 cells, consistent with the immunohistochemical staining of RAGE in vivo. CONCLUSION: VSMCs show very low levels of RAGE expression; thus, activation of VSMCs by AGEs does not occur. In diabetes, RAGE expression in VSM may increase to the extent that it becomes activated by AGEs in a manner that would contribute to the process of atherosclerosis. PMID- 16316351 TI - Effect of hereditary obesity on renal expressions of NO synthase, caveolin-1, AKt, guanylate cyclase, and calmodulin. AB - BACKGROUND: Obesity has emerged as a major cause of diabetes, cardiovascular disease, and renal insufficiency worldwide. Obese Zucker rats exhibit hyperphagia, obesity, insulin resistance, hyperlipidemia, and glomerulosclerosis and are frequently used as a model to study hereditary form of metabolic syndrome. Nitric oxide plays a major role in preservation of renal function and structure. The present study was designed to test the hypothesis that renal disease in this model may be associated with down-regulation of endothelial (eNOS) and neuromal NO synthases (nNOS) in the kidney. The study further sought to explore expressions of caveolin-1, phospho AKt, and calmodulin, which regulate activities of constituitive NOS isoforms, as well as soluble guanylate cyclase (sGC), which is involved in NO signaling. METHODS: Twenty-two-week-old male obese and lean Zucker rats were studied. Body weight, serum lipids, urine albumin excretion, and renal tissue abundance of the above proteins were determined. RESULTS: Serum glucose and arterial pressure were unchanged, whereas urinary NO metabolite (NO(chi)) excretion and renal tissue nitrotyrosine abundance were markedly reduced (denoting depressed NO production) in the obese versus lean Zucker rats. This was accompanied by significant glomerulosclerosis, tubulointerstitial damage, renal immune cell infiltration, marked down regulations of renal tissue eNOS and nNOS, mild reduction of caveolin-1, and unchanged calmodulin, phospho-AKt, and sGC. CONCLUSION: Hereditary obesity can result in down-regulations of kidney eNOS and nNOS, marked reduction of NO production, and glomerulosclerosis prior to the onset of frank diabetes and hypertension. PMID- 16316352 TI - Early detection of progressive renal dysfunction in patients with coronary artery disease. AB - BACKGROUND: An association between renal hemodynamic dysfunction and coronary artery disease (CAD) has been documented in chronic renal failure; however, no information is available in CAD patients with normal glomerular filtration rate (GFR). This study was aimed at evaluating early abnormalities and outcome of renal function in CAD patients. METHODS: In 15 nondiabetic patients with normal renal function and no significant stenoses in renal arteries, and having undergone coronary arteriography, we studied systemic and renal hemodynamics before and after a vasodilating stimulus induced by aminoacid (AA) infusion. A control group (C) consisted of 15 sex- and age-matched kidney donors. The statistical adequacy of the sample size was preliminarily verified. Renal clearances were repeated after two years. RESULTS: At baseline, GFR (mL/min/1.73 m2) averaged 81.4 +/- 3.8 in CAD and 83.7 +/- 1.4 in C (P= NS); RPF (mL/min/1.73 m2) was 297 +/- 22 in CAD and 456 +/- 15 in C (P < 0.0001); filtration fraction was higher in CAD (P < 0.001). Plasma renin activity was higher in CAD (P < 0.005). The number of coronary stenoses was inversely correlated with RPF but not with GFR. In CAD, at variance with C, AA did not induce any increment of GFR, while RPF increased without achieving the unstimulated value of C. Blood pressure was comparable in CAD and C at baseline and not modified by AA. After two years, a significant decrease in GFR (-14%, P < 0.001) and RPF (-15%, P < 0.001) occurred only in CAD, and in either group, the response to AA did not differ from that detected at baseline. CONCLUSION: In CAD patients with normal GFR, reduction in renal perfusion and absence of renal functional reserve likely represent early markers of progressive renal dysfunction. PMID- 16316353 TI - Quantitative estimation of renal blood flow by power Doppler ultrasonography in renovascular hypertensive dogs. AB - BACKGROUND: We estimated the value of power Doppler (PD) imaging analysis for the quantitative assessment of renal cortical blood flow (RCBF) in chronic two kidney, one-clip (2K-1C) hypertensive dogs. METHODS: To evaluate the correlation between RCBF and PD signals, RCBF and the mean pixel intensity (MPI) of PD signals were simultaneously obtained at same region in renal cortex under progressive constriction of left main renal artery in five mongrel dogs. RCBF was measured by electrolytic hydrogen gas clearance method, and PD images were transferred to computer and analyzed by the image-analysis software Openlab. To assess the value of quantitative PD imaging analysis on RCBF in renovascular hypertension, in six mongrel dogs with chronic 2K-1C hypertension, PD images in both of clipped kidneys (CK) and non-clipped kidneys (NK) were obtained and analyzed before and 60 minutes after the intravenous infusion of captopril or sodium nitroprusside at 10-minute intervals. RESULTS: There was a linear correlation between RCBF and MPI (r= 0.878, P < 0.0001). MPI in both CK and NK significantly increased after the infusion of captopril, while no significant change was observed in both CK and NK after the infusion of sodium nitroprusside, despite similar reduction of mean arterial blood pressure. CONCLUSION: Our data suggest that the acute inhibition of angiotensin-converting enzyme increased RCBF in both CK and NK of chronic 2K-1C dogs. The quantitative analysis of PD flow signals in kidney is noninvasive and a useful method to evaluate regional changes of renal tissue blood flow in various renal diseases. PMID- 16316354 TI - Role of AT-1 receptor in regulation of vascular MCP-1, IL-6, PAI-1, MAP kinase, and matrix expressions in obesity. AB - BACKGROUND: Metabolic syndrome has emerged as the major cause of atherosclerosis. The associated atherosclerosis is accompanied by and, in part, due to inflammation. In an attempt to explore the molecular sources of vascular inflammation and possible involvement of renin-angiotensin system, we studied obese Zucker rats, which exhibit all features of metabolic syndrome. METHODS: Seven-week-old male obese Zucker rats were randomized to losartan-treated (100 mg/L drinking H2O) and untreated groups. Lean Zucker rats served as controls. After four months, aortas were obtained and processed for various determinations by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot and immunohistochemical analysis for collagen type IV. RESULTS: Compared to the lean controls, obese Zucker rats showed significant increases in collagen staining, as well as expressions of collagen, fibronectin, plasminogen activator inhibitor-1, and two major proinflammatory mediators (i.e., interleukin-6 and monocyte chemoattractant protein-1). This was associated with significant increases in p38 and ERK1/2 mitogen activated protein kinase activities, as well as marked up regulation of angiotensin II type 1 receptor (AT-1R) mRNA expression. These abnormalities were prevented by administration of the AT-1R blocker (ARB). CONCLUSION: The untreated obese Zucker rats exhibit increased matrix protein accumulation in the aorta and marked up-regulations of proinflammatory and profibrotic pathways. These abnormalities are associated with up-regulation of AT 1R and are prevented by AT-1R blockade pointing to the potential role of AT-1R activation. PMID- 16316355 TI - Prealbumin, mortality, and cause-specific hospitalization in hemodialysis patients. AB - BACKGROUND: Prealbumin (transthyretin) is a hepatic secretory protein thought to be important in the evaluation of nutritional deficiency and nutrition support. Prior studies have suggested that the serum prealbumin concentration is independently associated with mortality in hemodialysis patients, even with adjustment for serum albumin and other nutritional parameters. METHODS: To determine whether prealbumin was independently associated with mortality and morbidity (cause-specific hospitalization) in hemodialysis patients, we analyzed data on 7815 hemodialysis patients with at least one determination of serum prealbumin during the last three months of 1997. Unadjusted, case mix-adjusted, and multivariable-adjusted relative risks of death were calculated for categories of serum prealbumin using proportional hazards regression. We also determined whether the prealbumin concentration was associated with all-cause, cardiovascular, infection-related, and vascular access-related hospitalization. RESULTS: The relative risk (RR) of death was inversely related to the serum prealbumin concentration. Relative to prealbumin > or =40 mg/dL, the adjusted RRs of death were 2.41, 1.85, 1.49, and 1.23 for prealbumin <15, 15-20, 20-25, and 25 30 mg/dL, respectively. The adjusted RRs of hospitalization due to infection were 2.97, 1.95, 1.81, and 1.61 for prealbumin <15, 15-20, 20-25, and 25-30 mg/dL, respectively. The adjusted RRs of vascular access-related hospitalization were 0.48, 0.52, 0.58, and 0.71 for prealbumin <15, 15-20, 20-25, and 25-30 mg/dL, respectively. While serum albumin was strongly associated with mortality and all cause hospitalization, it was not associated with hospitalization due to infection, and lower levels were associated with higher rather than lower rates of vascular access-related hospitalization. CONCLUSION: In hemodialysis patients, lower prealbumin concentrations were associated with mortality and hospitalization due to infection, independent of serum albumin and other clinical characteristics. Higher prealbumin concentrations were associated with vascular access-related hospitalization. In light of these findings, more intensive study into the determinants and biological actions of prealbumin (transthyretin) in end stage renal disease is warranted. PMID- 16316356 TI - Health-related quality of life and estimates of utility in chronic kidney disease. AB - BACKGROUND: Health-related quality of life and estimates of utility have been carefully evaluated in persons with end-stage renal disease. Fewer studies have examined these parameters in persons with chronic kidney disease (CKD). METHODS: To determine the relations among kidney function, health-related quality of life, and estimates of utility, we administered the Kidney Disease Quality of Life Short Form 36 (KDQOL-36), Health Utilities Index (HUI)-3, and Time Trade-off (TTO) questionnaires to 205 persons with CKD. Persons with CKD stages 4 and 5 (estimated GFR <30 mL/min/1.73 m2, N= 115) were tested two to eight times over the subsequent two years. The relations among estimated glomerular filtration rate (eGFR), and changes in health-related quality of life and utility over time were estimated using mixed effect regression models. Models were adjusted for age, sex, race, and diabetes. RESULTS: Mean scores on the KDQOL-36 generic components, HUI-3, and TTO suggested considerable loss of function and well-being in CKD relative to population norms. On cross-sectional analysis, lower levels of kidney function were associated with significantly lower scores on the SF-12 Physical Health Composite (P= 0.002), the Burden of Kidney Disease subscale (P < 0.0001), and the Effects of Kidney Disease subscale (P < 0.0001) of the KDQOL 36trade mark. Kidney function was significantly associated with the TTO (P= 0.008) and global HUI-3 utility (P= 0.016) although these associations were attenuated after adjustment for diabetes. A decline in eGFR was associated with a significant increase in the reported Burden of Kidney Disease (5.0 point change per year per mL/min/1.73 m2 decline in eGFR) and with marginally significant changes in the Dexterity and Pain attributes of the HUI-3. Mean HUI-3 scores for persons with CKD stages 4 and 5, absent dialysis, were in the range previously reported for persons with stroke and severe peripheral vascular disease. CONCLUSION: Health-related quality of life and estimates of utility are distressingly low in persons with CKD. Self-reported outcomes should be considered when evaluating health policy decisions that affect this population. PMID- 16316357 TI - Progressive accumulation of lanthanum in the liver of normal and uremic rats. AB - BACKGROUND: The loss of renal function in chronic kidney disease (CKD) patients leads to a variety of metabolic disturbances that promote the development of secondary hyperparathyroidism (SH) and bone disease. One of these alterations, hyperphosphatemia, is a persistent problem in patients with CKD and is an important factor in the development and severity of SH as well as coronary artery and cardiac valve calcification. To help control serum phosphorus (P), phosphate binders are used to minimize intestinal absorption of P in these patients. Since the use of aluminum- and calcium-based phosphate binders can result in severe side effects, other phosphate binders have been proposed. One of these is lanthanum (La) carbonate. METHODS: Since there is some controversy as to the degree La is absorbed by the intestine, whether it is deposited in tissues, and what effect its prolonged use would have in patients with chronic renal failure, we studied normal and uremic rats fed a control diet or a diet with La carbonate (1.5% elemental La) for 45, 90, or 110 days. RESULTS: As expected, plasma creatinine increased after the induction of uremia and continued to increase with time. At 45 and 90 days, dietary La produced a reduction in plasma P in normal rats (N + La) [45 days: 3.15 +/- 0.35 vs. normal control (NC) 4.41 +/- 0.15 mg/dL, P < 0.01; 90 days: 3.20 +/- 0.30 vs. 4.09 +/- 0.15 mg/dL, P < 0.05], but not at 110 days. Plasma P was initially lower (45 days) in both groups of La treated rats (U + La: 3.47 +/- 0.28 vs. UC: 4.91 +/- 0.27 mg/dL, P < 0.01), but was not at 90 and 110 days. Whole blood La content was not increased in rats fed the La diet and is, therefore, not a good indicator of the La content of tissue. La was increased in kidney, femur, and especially liver in normal and uremic rats fed the La diet. Uremia markedly enhanced tissue accumulation of La (liver, 110 days: N + La: 849 +/- 152 vs. U + La: 2676 +/- 479 ng/g wet wt, P < 0.01), which increased in a time-dependent manner. The increase in La in the liver of normal rats was 44-fold compared to control and 98-fold in the liver of uremic rats compared to uremic control. CONCLUSION: La accumulates in tissues, most strikingly in the liver, and renal failure accelerates the accumulation. While this was a relatively short-term study, La accumulation increased in a time dependent manner. The effect of long-term La treatment on patients with chronic renal insufficiency needs to be evaluated. PMID- 16316358 TI - Intravenous iron increases labile serum iron but does not impair forearm blood flow reactivity in dialysis patients. AB - BACKGROUND: There are concerns about adverse vascular effects of intravenous iron by inducing oxidative stress. We therefore examined the effect of a single high dose of intravenous iron on endothelial function and biochemical markers of iron homeostasis. METHODS: In a randomized, placebo-controlled, double-blind, parallel group study, forearm blood flow (FBF) was assessed by strain-gauge plethysmography in 38 peritoneal dialysis patients before and after a single intravenous infusion of 300 mg iron sucrose. RESULTS: Iron infusion increased total (Delta 601 microg/100 mL, CI 507, 696) and non-transferrin-bound iron (Delta 237.2 micromol/L, CI 173.6, 300.8) approximately 10-fold, as well as redox active iron nearly five-fold (Delta 0.76 micromol/L, CI 0.54, 0.98). After iron infusion basal FBF was 59% higher than after placebo. FBF response to acetylcholine before and after iron infusion was 263 +/- 32% and 310 +/- 33%, corresponding to 304 +/- 43% and 373 +/- 29% in the placebo group, respectively. Before and after iron or placebo infusion, glyceryl-trinitrate increased resting FBF to 232 +/- 22% and 258 +/- 21% in the iron group, and to 234 +/- 18% and 270 +/- 30% in the placebo group. L-N-monomethyl-arginine decreased FBF to 70 +/- 4% and 72 +/- 3% before and after iron, and to 74 +/- 4% and 73 +/- 4% before and after placebo infusions, respectively. Despite higher basal FBF after iron infusion, absolute and relative FBF changes in response to vasoactive substances were not significantly different between iron and placebo groups. CONCLUSION: Our data suggest that 300 mg intravenous iron sucrose has a vasodilatory effect, but does not impair vascular reactivity in dialysis patients, despite a significant increase in non-transferrin-bound and redox-active iron. PMID- 16316359 TI - Antiproteinuric effect of oral paricalcitol in chronic kidney disease. AB - BACKGROUND: Proteinuria is a marker of cardiovascular and renal disease in patients with chronic kidney disease (CKD), and reduction in proteinuria has been associated with improved cardiovascular and renal outcomes. While active vitamin D and its analogs have been shown to have renal protective effects in animals, these hormones have not been shown to reduce proteinuria in CKD patients. METHODS: In three double-blind, randomized, placebo-controlled studies to evaluate the safety and efficacy of oral paricalcitol, 220 CKD stage 3 and 4 patients with secondary hyperparathyroidism (SHPT) were randomized to oral paricalcitol (N= 107, mean dose 9.5 microg/week) or placebo (N= 113) and followed for up to 24 weeks. In conjunction with other safety measures, proteinuria was measured by dipstick and read by an automated reader at the beginning and end of trial. We subsequently analyzed the dipstick data to evaluate the effect of paricalcitol on proteinuria. RESULTS: At baseline, proteinuria was present in 57 patients randomized to oral paricalcitol and 61 patients randomized to placebo (NS). At the final visit, 29/57 (51%) of the paricalcitol patients compared to 15/61 (25%) placebo patients had reduction in proteinuria, P= 0.004 (odds for reduction in proteinuria 3.2 times greater for paricalcitol patients, 95% CI 1.5 6.9). For the patients who had both proteinuria at baseline and parathyroid hormone (PTH) suppression (end point defined as 2 consecutive > or =30% decreases in iPTH from baseline), 27/51 (53%) patients had a reduction in the proteinuria in the paricalcitol group and 0/7 (0%) had a reduction in proteinuria in the placebo group. Reduction of proteinuria favored patients on paricalcitol, regardless of age, sex, race, diabetes mellitus, hypertension, or use of therapies to block the renin-angiotensin-aldosterone system (RAAS). CONCLUSION: Our results demonstrate that the reduction in proteinuria was associated with paricalcitol treatment, and the reduction in proteinuria was independent of concomitant use of agents that block the RAAS. Paricalcitol as a potential pharmacologic means of reducing proteinuria in CKD patients warrants further investigation. PMID- 16316360 TI - Beneficial impact of spironolactone in diabetic nephropathy. AB - BACKGROUND: Aldosterone has been suggested to play a role in the initiation and progression of diabetic nephropathy. Currently recommended treatment with angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor blockers [renin-angiotensin system (RAS) blockade] does not suppress circulating aldosterone sufficiently. We therefore aimed to evaluate the short-term effect of aldosterone antagonism with spironolactone on albuminuria and blood pressure in diabetic nephropathy. METHODS: Twenty Caucasian type 1 diabetic patients with persistent macroalbuminuria despite antihypertensive treatment, including RAS blockade, completed this double-masked, randomized cross-over trial. Patients were treated in random order with spironolactone 25 mg once daily and matched placebo for two months, respectively, on top of usual antihypertensive treatment. After each treatment period albuminuria, 24-hour blood pressure, and glomerular filtration rate (GFR) were determined. RESULTS: Spironolactone on top of usual antihypertensive treatment induced a 30% (95% CI 17 to 41) reduction in albuminuria from [geometric mean (95% CI)] 831 (624 to 1106) mg/24-hour on placebo treatment (P < 0.001), and a reduction in fractional albumin clearance of 35% (20 to 46, P < 0.001). Twenty-four-hour blood pressure showed an insignificant reduction of [mean reduction (95% CI)] 8 (-1 to 17)/3 (-0.2 to 7) mm Hg (P < 0.10). There was an insignificant reversible reduction in GFR during treatment with spironolactone. On spironolactone treatment, one patient was excluded due to hyperkalemia (plasma potassium 5.7 mmol/L) and one due to orthostatic dizziness. Otherwise treatment was well tolerated. CONCLUSION: Our results suggest that spironolactone treatment on top of recommended antihypertensive treatment reduces blood pressure and may offer additional renoprotection in type 1 diabetic patients with diabetic nephropathy. PMID- 16316361 TI - Prevalence of decreased kidney function in Chinese adults aged 35 to 74 years. AB - BACKGROUND: Chronic kidney disease (CKD) is a major public health burden in Western countries but little is known about its impact in developing countries. We estimated the prevalence and absolute burden of CKD in the general adult population in China. METHODS: A cross-sectional survey was conducted in a nationally representative sample of 15,540 Chinese adults aged 35 to 74 years in 2000 and 2001. Serum creatinine was measured using the modified kinetic Jaffe reaction method at a central laboratory calibrated to the Cleveland Clinic Foundation laboratory. Glomerular filtration rate (GFR) was estimated using the simplified equation developed by the Modification of Diet in Renal Disease study. CKD was defined as an estimated GFR <60 mL/min/1.73m2. RESULTS: Overall, the age standardized prevalences of GFR 60 to 89, 30 to 59, and <30 mL/min/1.73m2 were 39.4%, 2.4%, and 0.14%, respectively, in Chinese adults aged 35 to 74 years. The overall prevalence of CKD (GFR <60 mL/min/1.73m2) was 2.53%, representing 11,966,653 persons (1.31% or 3,185,330 men and 3.82% or 8,781,323 women). The age specific prevalence of CKD was 0.71%, 1.69%, 3.91%, and 8.14% among persons 35 to 44, 45 to 54, 55 to 64, and 65 to 74 years old, respectively. The age standardized prevalence of CKD was similar in urban (2.60%) and rural (2.52%) residents but was higher in south China (3.05%) than in north China (1.78%) residents. CONCLUSION: Although the prevalence of CKD in China was relatively low, the population absolute burden is substantial. These data warrant a national program aimed at detection, prevention, and treatment of CKD in China. PMID- 16316362 TI - A randomized, controlled trial comparing IV iron sucrose to oral iron in anemic patients with nondialysis-dependent CKD. AB - BACKGROUND: Although iron deficiency frequently complicates anemia in patients with nondialysis-dependent CKD (ND-CKD), the comparative treatment value of IV iron infusion and oral iron supplementation has not been established. METHODS: In a randomized, controlled multicenter trial, we compared the efficacy of iron sucrose, given as 1 g in divided IV doses over 14 days, with that of ferrous sulfate, given 325 mg orally thrice daily for 56 days in patients with ND-CKD stages 3 to 5, Hb < or =11 g/dL, TSAT < or =25%, and ferritin < or =300 ng/mL. Epoetin/darbepoetin therapy, if any, was not changed for eight weeks prior to or during the study. RESULTS: The proportion of patients achieving the primary outcome (Hb increase > or =1 g/dL) was greater in the IV iron treatment group than in the oral iron treatment group (44.3% vs. 28.0%, P= 0.0344), as was the mean increase in Hb by day 42 (0.7 vs. 0.4 g/dL, P= 0.0298). Compared to those in the IV iron group, patients in the oral iron treatment group showed a greater decline in GFR during the study (-4.40 vs. -1.45 mL/min/1.73m2, P= 0.0100). No serious adverse drug events (ADE) were seen in patients administered IV iron sucrose as 200 mg IV over two to five minutes, but drug-related hypotension, including one event considered serious, occurred in two females weighing less than 65 kg after 500 mg doses were given over four hours. CONCLUSION: IV iron administration using 1000 mg iron sucrose in divided doses is superior to oral iron therapy in the management of ND-CKD patients with anemia and low iron indices. PMID- 16316363 TI - Effects of single-nucleotide polymorphisms in MTHFR and MTRR on mortality and allograft loss in kidney transplant recipients. AB - BACKGROUND: Plasma total homocysteine (tHcy) is associated with cardiovascular outcomes in kidney transplant recipients (KTR). The methylenetetrahydrofolate reductase (MTHFR) 677C>T polymorphism, an important determinant of plasma tHcy concentrations, could therefore constitute an important prognostic marker. METHODS: We prospectively followed 710 KTR over >6 years. The MTHFR677C>T, MTHFR1298A>C, MTHFR1793G>A, and MTRR66A>G polymorphisms were analyzed. Demographic, clinical, and transplant-related information was obtained, and patients were followed-up using the Austrian Dialysis and Transplant Registry. Using Cox regression, we established the independent relations of each genotype to the risk of death from any cause, and/or kidney allograft loss. RESULTS: During a median follow-up of 6.1 years, 154 participants died and 260 kidney allografts were lost. Compared to patients with the MTHFR677CC genotype, patients with MTHFR677CT had an adjusted relative mortality risk of 1.02 (95%CI 0.70 1.47), and those with MTHFR677TT of 0.98 (95%CI 0.52-1.85). Compared to MTHFR677CC, the relative risks of kidney allograft loss were 0.93 (95%CI 0.70 1.23; MTHFR677CT) and 0.78 (95%CI 0.47-1.30; MTHFR677TT), respectively. None of the other genotypes were associated with the risks studied, either. These findings did not depend on whether we controlled for tHcy levels. CONCLUSION: This study does not support the routine use of MTHFR or MTRR genotyping for prognostic evaluation or risk-stratification in kidney transplant recipients. PMID- 16316364 TI - Double target comparison of blood-side methods for measuring the hemodialysis dose. AB - BACKGROUND: Despite the fact that urea kinetic modelling has been successfully applied to quantify the hemodialysis since the beginning of the 1980s, there is not a consensus yet concerning which is the most proper dialysis dose index and the method for calculating it. In this work, we propose that a combined measurement of the dialysis dose from two complementary perspectives of the removal process should provide a more complete description of dialysis than a measurement alone. This hypothesis is reviewed and the measuring methods are compared. METHODS: A cross-sectional randomized clinical study over 98 stable ESRD patients submitted to thrice-weekly hemodialysis was carried out with the aim of comparing 16 blood-side methods for measuring the hemodialysis dose from patient and dialyzer perspectives. The availability of urea rebound measurements and computational resources have been taken into account. RESULTS: The outcomes point to four novel blood-side methods as the most accurate for measuring the effective dialysis system Kt/V (mKt/V) in clinical conditions. Their limits of agreement (mean +/- 2.SD) range from 1.93 +/- 2.09% for a non-iterative method without the urea rebound measurement (BUN3) to -0.08 +/- 0.58% for an iterative method with BUN3. The best non-iterative blood-side method for measuring the equilibrated Kt/V (eKt/V) is the second generation formula of Daugirdas (-2.42 +/ 1.05%) when BUN3 is available and the rate equation of Daugirdas and Schneditz ( 1.74 +/- 7.91%) when BUN3 is not available. The difference mKt/V-eKt/V is significant and positive, and increases with the dialysis dose in a personalized manner. CONCLUSION: We have confirmed the arguments that support the hypothesis of the study. The best blood-side methods for the combined measurement of dialysis dose as a function of the available resources have been determined. PMID- 16316365 TI - Acute plasma ghrelin and leptin responses to oral feeding or intraperitoneal hypertonic glucose-based dialysate in patients with chronic renal failure. AB - BACKGROUND: Chronic renal failure (CRF) is associated with increased plasma levels of ghrelin and leptin, but the regulation of the secretion of these hormones has been insufficiently studied, in this setting. The aim of this study was to analyze the acute effects of oral feeding or intraperitoneal 3.86% glucose based dialysate infusion on plasma ghrelin and leptin levels in patients with CRF undergoing peritoneal dialysis (PD). METHODS: Following a crossover design, 10 patients and eight healthy controls underwent a standardized oral intake, a 3.86% glucose-based dialysate PD exchange (patients) and placebo oral intake. We scrutinized acute changes in plasma ghrelin, leptin, glucose, insulin, and growth hormone (GH) levels. RESULTS: In patients, total ghrelin decreased modestly immediately after oral feeding (nadir 90.6% of baseline, range 85.1, 94.5, P= 0.03) or the PD exchange test (92.2%, range 58.7, 101.9, P= 0.05) (median). Response to oral feeding was markedly blunted when compared with healthy individuals (73.8%, range 56.1, 89.1, P= 0.007) (P < 0.005 vs. patients). Plasma acyl-ghrelin had a less marked but more persistent decay after the PD exchange test (nadir 80.4%, range 55.1, 96.3, P= 0.02) than after oral intake (64.4%, range 45.6, 82.3, P= 0.005); again, changes were more intense in normal controls (47.4%, range 32.1, 67.3, P= 0.01) (P < 0.05 vs. patients). Leptin levels decreased slightly (P < 0.05) after the PD exchange in patients, but did not respond acutely to oral feeding in patients or controls. CONCLUSION: Ghrelin secretion is partially refractory to the acute inhibitory effect of oral feeding in patients with CRF undergoing PD therapy. A 3.86% glucose-based PD exchange results in a significant decrease of plasma ghrelin levels. Plasma leptin levels are not acutely affected by oral feeding in patients with CRF or healthy individuals. PMID- 16316366 TI - Outcomes of tunneled femoral hemodialysis catheters: comparison with internal jugular vein catheters. AB - BACKGROUND: Tunneled femoral vein dialysis catheters are used as a last resort when all other options for a permanent vascular access or thoracic central vein catheter have been exhausted. There is little published literature on the complications or outcomes of tunneled femoral catheters. METHODS: Using a prospective, computerized vascular access database, we identified all tunneled femoral dialysis catheters placed at the University of Alabama at Birmingham during a five-year period. The clinical features, catheter patency, and complications in these patients were compared to those observed in a group of sex , age-, and date-matched control patients with tunneled internal jugular vein dialysis catheters. RESULTS: During the study period, 27 patients received a tunneled femoral dialysis catheter, accounting for 1.9% of all tunneled catheters placed. Only 7 patients (26%) were able to convert to a new permanent dialysis access. The primary catheter patency (time from placement to exchange) was substantially shorter for femoral catheters than for internal jugular dialysis catheters (median survival, 59 vs. >300 days, P < 0.0001). Infection-free survival was similar for both groups (P= 0.66). Seven patients with femoral catheters (or 26%) developed an ipsilateral deep vein thrombosis, but catheter use was possible with anticoagulation. CONCLUSION: Tunneled femoral dialysis catheters have a substantially shorter primary patency, but a similar risk of catheter-related bacteremia, as compared with internal jugular vein catheters. An ipsilateral lower extremity deep vein thrombosis occurs commonly after placement of a femoral dialysis catheter, but does not preclude continued catheter use. PMID- 16316367 TI - Adventitial remodeling with increased matrix metalloproteinase-2 activity in a porcine arteriovenous polytetrafluoroethylene grafts. AB - BACKGROUND: We hypothesized the source of early proliferating cells contributing to venous stenosis formation in a porcine hemodialysis grafts is the adventitia and media, and migration of these cells is greatest within the first two weeks following graft placement, resulting in increased matrix metalloproteinase-2 (MMP 2) activity. METHODS: Polytetrafluoroethylene grafts from the iliac artery to the ipsilateral iliac vein were placed in 23 pigs and 5-Bromo-2'-deoxyuridine (BrdU) was given at 24 and 48 hours after surgery to assess cell proliferation and migration. Angiography and magnetic resonance angiography was performed. Animals were euthanized on day three (N= 6), day seven, (N= 5), day 14 (N= 6), and days 19 to 26 (N= 6) after graft placement, and stenotic tissue and unaffected contralateral iliac vein were removed for zymography and immunostaining. RESULTS: Migration of cells derived from the adventitia and media peaked at day 14. Adventitial diameter of the stenotic vein decreased, while the intima to media ratio increased. MMP-2 activity peaks at day seven in the adventitia and days 19 to 26 in the intima. CONCLUSION: These results confirm our hypothesis that the source of cells resulting in venous stenosis formation is derived from the adventitia and media, with cell migration being greatest within the first two weeks after graft placement with translocation of these cells into the intima at four weeks. MMP-2 activity peaks at day seven in the adventitia and again at days 19 to 26 in the intima. A key to limiting venous stenosis formation may lie in inhibiting MMP-2 by adventitial and medial targeting. PMID- 16316368 TI - Is there a pharmacologic basis for combination renin axis blockade? PMID- 16316369 TI - The response to loss of function of one kidney: it all depends upon what is happening next door. PMID- 16316371 TI - The need for contamination control in studies on lanthanum biodisposition. PMID- 16316372 TI - Lanthanum pharmacokinetics: are rat data misleading? PMID- 16316375 TI - The need for careful interpretation of animal data on lanthanum. PMID- 16316376 TI - Consideration of use of neuraminidase inhibitors such as oseltamivir and zanamivir in IgA nephropathy. PMID- 16316377 TI - Thrombocytopenia induced by nicotinamide in hemodialysis patients. PMID- 16316378 TI - Muscle IGF-I levels in hemodialysis patients. PMID- 16316379 TI - Anti-proteinase 3 antibody binding to neutrophils as demonstrated by confocal microscopy. PMID- 16316380 TI - Daily dialysis, nocturnal dialysis, and randomized controlled trials: are we asking the right questions? PMID- 16316382 TI - Serum fetuin-A and aortic calcification in hemodialysis patients. PMID- 16316385 TI - The global challenge of chronic kidney disease. PMID- 16316388 TI - Proceedings of International Parvovirus Meeting 2005. July 15-16, 2005. Leipzig, Germany. PMID- 16316389 TI - An annotated historical account of canine parvovirus. AB - A brief annotated history of canine parvovirus-type 2 (CPV-2) and its variants is summarized with emphasis on the most significant contributions of individuals involved in the initial recognition of CPV-2 and subsequent discoveries that have advanced our knowledge of the nature and evolution of this novel canine virus. Time has obscured the observations of many veterinary clinicians and researchers throughout the world who sensed the presence of a new disease when CPV-2 first made its appearance in 1978 and then, within 1-2 years, spread worldwide. Since 1979, nearly 600 articles, papers, numerous text chapters and monographs have been published on the subject of CPV-2. The early history is well known by veterinary infectious diseases specialists and noteworthy publications are recorded on the National Library of Medicine (USA) website, PubMed and in review articles. Because of the great number of publications, it is not practicable to cite them individually; however, reference is made to certain individuals, reviews and selected papers that I consider particularly relevant to the history of progress in the understanding of CPV-2 and the disease it causes. The clinical disease caused by CPV-2 and its variants, the immune response to infection or vaccines, host range and the development of practical diagnostic assays are noted in historical context. The basic biological properties and the physical, molecular and antigenic structure of CPV-2 and its variants are also discussed briefly. Finally, key players who have contributed to the antigenic and DNA sequence (evolutionary) relationships between CPV-2 and the other autonomous parvoviruses of carnivores are noted and hypotheses regarding the origin and evolution of CPV-2 and its variants are mentioned. PMID- 16316390 TI - Surveillance activity for canine parvovirus in Italy. AB - Recent identification of unusual canine parvovirus (CPV) mutants in cats and dogs suggests that CPV type 2 (CPV-2), which emerged suddenly in the late 1970s, is undergoing continual genetic and antigenic variations. A peculiarity of parvoviruses is that single-nucleotide substitutions may determine drastic phenotypic changes. The effects of either natural or artificial mutations on CPV phenotypic properties have been largely investigated, and this sets up CPV as an interesting model to study virus evolution. By monitoring the evolution of CPV-2 in Italy, we observed the onset and quick spread of a Glu-426 mutant, antigenically different from the pre-existing variants that were partially displaced within a few years of the initial identification of the new mutant. The identification of CPV-2 variants raises several questions concerning their impact on the efficacy of the current CPV-2 vaccines, based on the original CPV-2 strain that no longer exists in the field. PMID- 16316391 TI - New approaches for the molecular characterization of canine parvovirus type 2 strains. AB - Characterization of the canine parvovirus type 2 (CPV-2) is sometimes ambiguous, frequently requiring more than one technique for definitive prediction of the viral type. Taking into account the single-nucleotide polymorphisms encountered in the VP2-protein gene between types 2a and 2b and between type 2b and Glu-426 mutant (type 2c), two different minor groove binder (MGB) probe assays were developed for rapid identification of the CPV-2 variants. A total of 315 samples collected from dogs with diarrhoea were screened for CPV-2 by a real-time polymerase chain reaction (PCR) assay capable of detecting all CPV-2 types. In order to compare the type-specific assays with the traditional techniques [haemagglutination inhibition with monoclonal antibodies, PCR-restriction fragment-length polymorphism (RFLP), sequence analysis] for prediction of CPV-2 antigen specificity, the 203 samples tested CPV-2 positive were analysed using the different methods. The results showed a 100% concordance between the MGB probe assays and the combined conventional methods, with 116 samples characterized as type 2a, 32 as type 2b and 55 as type 2c. Therefore, the MGB probe assays represent a quick, reliable tool for prediction of CPV-2 antigen specificity, with regard to the more time-consuming assays currently used. PMID- 16316392 TI - Antibody levels and protection to canine parvovirus type 2. AB - The relationship between maternally derived antibody (MDA) levels and protection to canine parvovirus (CPV) infection in pups is reported. Twelve pups with a wide range of haemagglutination inhibiting (HI) titres of MDA to CPV were divided into four groups, with each group balanced for antibody titres. The dogs were inoculated with a field CPV-2b strain and clinical signs, virus shedding and antibody response were assessed. The CPV was not detected in the faeces of dogs with HI titres of 320 at any time. In dogs with HI titres up to 160, active CPV replication after challenge was demonstrated by real-time polymerase chain reaction. The successful infection of dogs with HI titres of 80 and 160 was confirmed by seroconversion, evaluated at day 14 post-infection. These findings demonstrated that CPV infection could also occur in the presence of MDA HI titres (> or =80) usually considered fully protective. PMID- 16316393 TI - Tissue distribution of two field isolates and two vaccine strains of porcine parvovirus in foetal organs after experimental infection of pregnant sows as determined by real-time PCR. AB - The aim of this study was to investigate the tissue distribution of two different field isolates and two vaccine strains of porcine parvoviruses (PPV) in infected piglets after transplacental infection. The viral load in 10 different foetal organs was determined by real-time polymerase chain reaction assays with SYBR Green targeting the viral VP2 gene and the genomic c-myc gene in 12 foetuses. The viral load in foetal tissues differed greatly among the different parvoviruses. Between one virulent field isolate compared with the other field isolate and the vaccine strains, the detected viral copy number differed in an order of magnitude of 10(9). The virulent isolate contained PPV in all 10 organs with viral loads varying between 10(11) and 10(15) per 10(6) cells. Concerning the other field isolate and the two vaccine strains, if PPV was detected, in most of the cases the highest viral load was found in foetal kidneys with a maximum viral load of 10(3) per 10(6) cells. Additionally, PPV was found in the heart of one foetus, in the liver and duodenum of one foetus and in the thymus of one foetus with viral loads varying between 10(2.1) and 10(3.5) per 10(6) cells. In completely mummified foetuses with no discriminable organs of foetuses infected with the vaccine strains and the less virulent isolate, PPV was present in very low amounts or even below the detection limit. PMID- 16316394 TI - Oncolytic potential of rodent parvoviruses for cancer therapy in humans: a brief review. AB - Summary Rodent parvoviruses are promising candidates for oncolytic virotherapy of cancer in humans because of their oncotropism (preferential killing of transformed cells) in the absence of pathogenicity. Here, we give an overview concerning the possible application of parvovirus H-1 for cancer therapy, with specific emphasis on malignant brain tumours in humans. PMID- 16316395 TI - Pathogenesis of aleutian mink disease parvovirus and similarities to b19 infection. AB - Aleutian mink disease parvovirus (ADV) is an unusual member of the autonomous parvoviruses in both its replication and pathogenesis. Infection of newborn mink kits results in an acute disease typified by virus replication in type II pneumocytes in the lung. This replication is permissive and cytopathic, characterized by the production of high levels of viral replicative intermediates and infectious progeny. However, infection of adult Aleutian mink leads to a chronic form of the disease termed Aleutian disease (AD). In this case, virus replication occurs predominantly in lymph node macrophages and is restricted, with viral DNA replication, RNA transcription, protein expression and production of infectious progeny occurring at low levels. B19 is the only autonomous parvovirus known to infect humans. The primary site of virus replication in both children and adults is in erythrocyte precursors in the blood and bone marrow, although viral genomes have been detected in various other tissues. B19 infection often causes a self-limiting disease although persistent infection of B19 can occur in both immuno-compromised and -competent people. Perhaps the most striking similarity between infection with ADV or with B19 is the important role the humoral immune response to infection has in pathogenesis. It can be both protective and pathogenic. Due to of the central role of antibody in the disease caused by either virus, understanding the specific roles of antibody production in protection, antibody-mediated enhancement of infection, the establishment of persistent infection and immune-mediated pathology will provide insight into the pathogenesis of these infections. A second similarity between the two viruses is the ability to establish persistent infection. Persistence of ADV is associated with restricted replication. Although many cellular factors may contribute to restricted virus replication, the interactions between the major non-structural protein, NS1, and the cells are likely to be critical. Parallels exist between the expression and post-translational modification of ADV and B19 NS1 proteins that may contribute to restriction of virus replication. Thus, a study of the regulation of NS1 expression and its interactions with cell signalling pathways may lead to increased understanding of the restricted replication of these two viruses, and perhaps of persistent infection. PMID- 16316396 TI - Pathogenesis of parvovirus B19 infection: host gene variability, and possible means and effects of virus persistence. AB - Since conducting follow-up studies of patients with acute symptomatic parvovirus B19 infection which showed that a significant proportion of patients develop prolonged arthritis and chronic fatigue syndrome (CFS), we have become interested in the mechanisms of this phenomenon. We showed that these cases have high levels of pro-inflammatory cytokines in their circulation and that this correlates with the symptoms. However, the underlying mechanisms were not apparent, and we have used various approaches to begin studying this phenomenon. DNA polymorphisms were looked for and several were shown to be more common in these subjects compared with controls; these occur within genes of both the immune response [human leucocyte antigen (HLA)-DRB1, HLA-B, transforming growth factor (TGF)-beta1] and those involved in several other cellular functions (predominantly the cytoskeleton and cell adhesion). Interestingly, one particular single-nucleotide polymorphism (SNP) which is associated with symptomatic B19 infection occurs in the Ku80 gene which has recently been shown to be a B19 co-receptor. B19 persistence is probably the key to this phenomenon, and some new data are presented on short regions of sequence homology (17-26 bp) between human, mouse and rat parvoviruses and their respective hosts which occur in many host genes. This homology may provide a foothold for virus persistence and may also play a role in the genesis of disease through gene disruption. Finally, we used microarrays and TaqMan real-time polymerase chain reaction in 108 normal persons to study human gene expression in persons who are B19-seropositive versus B19 seronegative (age- and sex-matched) to examine the hypothesis that gene regulation may be altered in subjects harbouring the B19 virus DNA. Six genes were found to be differentially expressed with roles in the cytoskeleton (SKIP, MACF1, SPAG7, FLOT1), integrin signalling (FLOT1, RASSF5), HLA class III (c6orf48), and tumour suppression (RASSF5). These results have implications not only for B19 but also for other persistent viruses as well and confirmation is required. In conclusion, these disparate findings contribute to our understanding of the pathogenesis of B19 disease. We are using these studies as a starting point to study the phenomenon of chronic immune activation following B19 infection. PMID- 16316397 TI - Human parvovirus B19: a new emerging pathogen of inflammatory cardiomyopathy. AB - The human parvovirus B19 (PVB19), an erythrovirus causing diverse clinical manifestations ranging from asymptomatic or mild to more severe outcomes such as hydrops fetalis, is the only known human pathogenic parvovirus so far. Although enteroviruses have long been considered the most common cause of inflammatory cardiomyopathy, PVB19 is emerging as a important candidate. Recent studies have indicated an association of PVB19 with paediatric and adult inflammatory cardiac disease. However, whether or not PVB19 has an impact on inflammatory cardiomyopathy in adult patients is still unclear. The first hints for a possible aetiopathogenetic role of the PVB19-infection and the development of cardiac dysfunction were demonstrated by molecular biology utilizing in situ hybridization (ISH) and polymerase chain reaction (PCR). According to available evidence, PVB19-associated inflammatory cardiomyopathy is characterized by infection of endothelial cells of small intracardiac arterioles and venules, which may be associated with endothelial dysfunction, impairment of myocardial microcirculation, and penetration of inflammatory cells into the myocardium. PMID- 16316398 TI - Pathophysiology and aetiological diagnosis of inflammatory myocardial diseases with a special focus on parvovirus B19. AB - Inflammatory processes induced by viral or bacterial infections are believed to be one of the major pathogenetic mechanisms in myocardial diseases. Although the reason for progression to myocardial failure is not fully understood, postulated mechanisms include persistent viral infection alone or in combination with autoimmune processes. A variety of cardiotropic viruses have been identified to elicit myocarditis, with enteroviruses and adenoviruses as the most frequent causative agents in children and adolescents. However, parvovirus B19 (PVB19) has recently emerged as another potential pathogen in adult patients associated with inflammatory heart disease. Many dimensions of inflammatory heart disease coexist while different phases of the disease progress simultaneously: phase 1 is dominated by viral infection, phase 2 by the onset of (probably) multiple autoimmune reactions, and phase 3 by the progression to cardiac dilatation without the role of an infectious agent and cardiac inflammation. Taking these mechanisms into account, screening for viral and bacterial genome by polymerase chain reaction (PCR) and detection of inflammatory infiltrates by immunohistochemistry are considered crucial for establishing an aetiological diagnosis, thereby allowing initiation of specific therapeutic strategies. In a large cohort of 3345 consecutive patients with left ventricular dysfunction evaluated over a period of 10 years, prevalence of PVB19, coxsackievirus (CVB), human cytomegalovirus (HCMV), influenza A virus and adenovirus (ADV) genome was assessed by PCR. Inflammatory infiltrates within the myocardium were detected by immunohistochemistry according to the WHF criteria and by histopathology according to the Dallas criteria of myocarditis. For control, endomyocardial samples of patients with arterial hypertension were studied. Parvovirus B19 was the most often detected virus in all patient subgroups, with positivity ranging from 17% to 33%. Except for PVB19, CVB RNA (3%), ADV (2%) and CMV (3.9%) were the most frequently detected viral genomes. Interestingly, detection of PVB19 genome was significantly correlated with inflammatory heart disease and reduced ejection fraction. Importantly, an aetiological diagnosis requires the immunohistochemical and molecular biological investigation of endomyocardial biopsies. Such an approach may change the management of these diseases in the future. One of the aims of the study was to reveal the underlying dominant pathophysiological mechanisms in a for deciding on the most approriate therapy. PMID- 16316399 TI - Parvovirus B19 VP2-proteins produced in Saccharomyces cerevisiae: comparison with VP2-particles produced by baculovirus-derived vectors. AB - The capsids of human parvovirus B19 consist of two structural proteins, the minor capsid protein VP1 and the major-capsid protein VP2. The latter which constitutes for 95% of the capsid are able to form virus-like particles (VLPs) in yeast without the presence of VP1-proteins. VP2-proteins produced in Saccharomyces cerevisiae have the capacity to form VLPs in the absence of VP1-proteins. These yeast-derived VLPs resemble native virus or recombinant VP2-VLPs produced by baculovirus systems in respect of size, molecular weight and of antigenicity as shown by antigen-capture ELISA and T-cell proliferation tests. Regarding costs, yield and ease of handling particle production in yeast represents an alternative to the recombinant baculovirus expression system which is so far the source for VP2-VLPs of human parvovirus B19. PMID- 16316400 TI - Human parvovirus B19 infection and antiphospholipid-syndrome: the two sides of one medal? PMID- 16316401 TI - CD4(+) T-cell responses against the VP1-unique region in individuals with recent and persistent parvovirus B19 infection. AB - To date cellular immune responses against parvovirus B19 (B19) have not been studied extensively. The aim of this study was to examine the T-cell response against the VP1-unique region as the immunodominant part of the viral structural protein VP1 in individuals with different courses of B19 infection. Therefore, a group of 13 parvovirus-positive probands was separated into subgroups characterized for recent or acute, past or persistent infection by means of the presence of specific immunoglobulin (Ig)M and IgG isotypes and of viral DNA in blood and tissue. Transiently transfected B-cells expressing VP1-unique region were used in ELISpot assays to investigate T-cell responses directed against the VP1-unique region in peripheral blood mononuclear cells (PBMC) of individual donors. Significant numbers of interferon-gamma (IFN-gamma) secreting lymphocytes were detectable in PBMC of all individuals with recent, acute or persistent B19 infection, but not in PBMC of donors with past B19 infection and seronegative individuals. A more detailed analysis of IFN-gamma producing cells by intracellular cytokine staining by flow cytometry revealed, that CD4(+) T cells but not CD8(+) cytotoxic lymphocytes (CTL) were the major subpopulation of IFN gamma producing cells. These data strongly suggest the need of virus protein production for the maintenance of VP1-unique region-specific CD4(+) T-helper cell responses in B19-infected individuals. PMID- 16316402 TI - Human parvovirus B19 serology and avidity using a combination of recombinant antigens enables a differentiated picture of the current state of infection. AB - In order to improve serodiagnostic methods for the determination of the state of human parovirus B19 infection, a new test system, recomLine Parvovirus, based on the use of recombinant antigens, has been developed and evaluated. The test system combines the advantages of enzyme-linked immunosorbent assay (ELISA) methods with those of the Western blot technique. For the recombinant line assay, five antigens of human parvovirus B19 that were recombinantly produced in Escherichia coli were applied directly on nitrocellulose membranes: VP2, the aminoterminal and the carboxyterminal domain of VP1 (VP-N and VP-C), VP-1S another fragment of VP-N and NS1. In addition, empty virus particles isolated from eukaryotic cell cultures were also applied. The recombinant-line assay was used to detect human IgG and IgM antibodies directed against human parvovirus B19. In addition, the avidity of the IgG antibodies was investigated. The recombinant line assay was evaluated using 87 human serum samples of patients recently infected with human parvovirus B19 including 10 samples of three infection time courses and 100 serum samples of healthy blood donors. All results were compared with commercially available ELISAs. In the case of discrepancies, Western blot analysis was performed. The data revealed the recombinant line assay to be highly sensitive and specific. The individual determination of the human immune response against several recombinant antigens covering the structural proteins of human parvovirus B19 gives a deeper insight into the actual status of infection. In addition, the determination of IgG avidity against these individual recombinant antigens enables a more precise and differentiated picture of the infection event. PMID- 16316403 TI - GATA factors as key regulatory molecules in the development of Drosophila endoderm. AB - Essential roles for GATA factors in the development of endoderm have been reported in various animals. A Drosophila GATA factor gene, serpent (srp, dGATAb, ABF), is expressed in the prospective endoderm, and loss of srp activity causes transformation of the prospective endoderm into ectodermal foregut and hindgut, indicating that srp acts as a selector gene to specify the developmental fate of the endoderm. While srp is expressed in the endoderm only during early stages, it activates a subsequent GATA factor gene, dGATAe, and the latter continues to be expressed specifically in the endoderm throughout life. dGATAe activates various functional genes in the differentiated endodermal midgut. An analogous mode of regulation has been reported in Caenorhabditis elegans, in which a pair of GATA genes, end-1/3, specifies endodermal fate, and a downstream pair of GATA genes, elt-2/7, activates genes in the differentiated endoderm. Functional homology of GATA genes in nature is apparently extendable to vertebrates, because endodermal GATA genes of C. elegans and Drosophila induce endoderm development in Xenopus ectoderm. These findings strongly imply evolutionary conservation of the roles of GATA factors in the endoderm across the protostomes and the deuterostomes. PMID- 16316404 TI - Further extension of mammalian GATA-6. AB - Mammalian GATA-6, which has conserved tandem zinc fingers (CVNC-X(17)-CNAC)-X(29) (CXNC-X(17)-CNAC), is essential for the development and specific gene regulation of the heart, gastrointestinal tract and other tissues. GATA-6 recognizes the (A/T/C)GAT(A/T)(A) sequence, and interacts with other transcriptional regulators through its zinc-finger region. The mRNA of GATA-6 uses two Met codons in frame as translational initiation codons, and produces L- and S-type GATA-6 through leaky ribosome scanning. GATA-6 is subjected to cAMP-dependent proteolysis by a proteasome in a heterologous expression system. These protein-based characteristics of GATA-6 will be helpful for the identification of target genes, together with determination of the in vivo binding sites for GATA-6 and understanding of the complex network of gene regulation mediated by GATA-6. PMID- 16316405 TI - Molecular mechanisms for thyroid hormone-induced remodeling in the amphibian digestive tract: a model for studying organ regeneration. AB - During amphibian metamorphosis the digestive tract is extensively remodeled under the control of epithelial-connective tissue interactions. At the cellular level, larval epithelial cells undergo apoptosis, while a small number of stem cells appear, actively proliferate, and then differentiate to form adult epithelium that is analogous to its mammalian counterpart. Therefore the amphibian digestive tract is a unique model system for the study of postembryonic organ regeneration. As amphibian intestinal remodeling can be triggered by thyroid hormone (TH), the molecular mechanisms involved can be studied from the perspective of examining the expression cascade of TH response genes. A number of these genes have been isolated from the intestine of Xenopus laevis. Recent progress in the functional analysis of this cascade has shed light on key molecules in intestinal remodeling such as matrix metalloproteinase-11, sonic hedgehog, and bone morphogenetic protein-4. These genes are also thought to play key roles in organogenesis and/or homeostasis in both chick and mammalian digestive tract, suggesting the existence of conserved mechanisms underlying such events in terrestrial vertebrates. In this article, we review our recent findings in this field, focusing on the development of adult epithelium in the X. laevis intestine. PMID- 16316406 TI - Hairy is a cell context signal controlling Notch activity. AB - Notch signaling regulates cell fate determination and many developmental processes. Here we report that lateral inhibition, a major mechanism for Notch activity, is modulated by Hairy, a bHLH-WRPW protein. In Xenopus, Notch can have from inhibitory, permissive to enhancing roles in muscle or neural differentiation. These cell context-dependent effects correlate with Hairy expression levels from high to low, respectively, in the cells. Moreover, Notch effects can be altered upon manipulation of Hairy expression. We propose that Hairy provides a cell context in which a cell can interpret Notch and other extrinsic signals by controlling responsiveness of its target genes; this mode of Hairy-Notch interaction may apply in other systems. PMID- 16316407 TI - Pax3 transcripts in melanoblast development. AB - The transcription factor encoded by PAX3 is among the first expressed in the embryo, with a key role in development of the melanocytic lineage. Re-expression of PAX3, consistently observed in cutaneous malignant melanoma (CMM) as compared to normal melanocytes, appears linked to progression of CMM. Previous research has identified PAX3d (encoded by exons 1-9) as the predominant isoform present in CMM, together the with an alternate isoform PAX3c (encoded by exons 1-8). We investigated the expression of Pax3c and Pax3d transcripts during mouse development. The reverse transcription-polymerase chain reaction and immunohistochemistry experiments presented here implicate these transcripts in melanoblast development and demonstrate significant spatial and temporal differences in their expression. Differences in expression were also noted during active hair regrowth in adult skin, which is accompanied by proliferation and migration of melanoblasts into the hair cortex to color new hair. Results indicate that the defined spatial and temporal expression of Pax3d may be linked to either melanoblast proliferation or migration during melanogenesis. PMID- 16316408 TI - Expression and phosphorylation of TOPK during spermatogenesis. AB - Among normal organs and tissues, the MAPKK-like mitotic protein kinase TOPK is expressed exclusively in the testis. We analyzed the expression and phosphorylation of TOPK to address the functional role of this kinase during spermatogenesis. TOPK protein is expressed mainly in the cytosol of spermatocytes and spermatids, but not in spermatids and spermatogonia in situ. TOPK-Thr-9, a cdk1/cyclin B target residue, was specifically phosphorylated during mitotic and meiotic phases, while TOPK-Thr-198, a key amino acid for the ATP pocket, was constantly phosphorylated irrespective of the cell cycle. These data indicate that spermatogenic germ cells with vital proliferation activity express TOPK. As TOPK-Thr-9 was phosphorylated during both mitosis and meiosis, TOPK was indicted to play a role in cytokinesis and/or chromosomal segregation but not in DNA replication. PMID- 16316409 TI - A novel function of differentiation revealed by cDNA microarray profiling of p75NTR-regulated gene expression. AB - The expression of the p75 neurotrophin receptor (p75NTR) is diminished in epithelial cells during progression of prostate cancer in vivo and in vitro. Previous studies have demonstrated a role for p75NTR as a tumor suppressor in prostate growth. To better understand the molecular mechanism of p75(NTR) on tumor suppression, we utilized a complementary deoxyribonucleic acid microarray composed of approximately 6,000 human cancer-related genes to determine the gene expression pattern altered by re-introduction of p75NTR into PC-3 prostate tumor cells. Comparison of the transcripts in the neo and p75NTR-transfected cells revealed 52 differentially expressed genes, of which 21 were up-regulated and 31 were down-regulated in the presence of p75NTR. Based on the known biological functions of the p75NTR-regulated genes, we observed that p75NTR modulated the expression of genes that are critically involved in the regulation of differentiation as well as cell adhesion, signal transduction, apoptosis, tumor cell invasion, and metastasis. Several differentially expressed genes identified by microarray were selected for confirmation using quantitative real-time polymerase chain reaction. Immunoblot analysis further confirmed increased cellular retinoic acid-binding protein I (CRABPI) and IGFBP5 protein levels and decreased level of PLAUR protein with increasing p75NTR protein expression. As CRABPI was elevated far more than any other genes, we observed that the retinoids, all-trans retinoic acid and 9-cis retinoic acid, that bind CRABPI, promoted nitroblue tetrazolium-associated functional cell differentiation in p75NTR PC-3 cells, but not in neo control PC-3 cells. Subsequent examination of the retinoic acid receptors (RARs) expression levels demonstrated an absence of RAR-beta in the neo control cells and re-expression in the p75NTR expressing cells, consistent with previous findings where RAR-beta is believed to play a critical role as a tumor suppressor gene that is lost during de-differentiation of prostate epithelial cells. Whereas the RAR-alpha and -gamma protein levels remained unchanged, retinoid X receptor (RXR)-alpha and -beta also exhibited increasing protein levels with re-expression of the p75NTR protein. Moreover, the ability of p75NTR siRNA to knockdown levels of RAR-beta, RXR-alpha, and RXR-beta supports the specificity of the functional involvement of p75NTR in differentiation. Hence, re-expression of the p75NTR appears to partially reverse de-differentiation of prostate cancer cells by up-regulating the expression of CRABPI for localized sequestration of retinoids that are available to newly up regulated RAR-beta, RXR-alpha, and RXR-beta. PMID- 16316410 TI - Negative regulation of Gli1 and Gli2 activator function by Suppressor of fused through multiple mechanisms. AB - During animal development, the Hedgehog (Hh) signal transduction pathway plays critical roles in cell fate determination and tissue patterning. In humans, aberrant Hh signaling has been linked to several genetic disorders and cancers. Binding of Hh to its receptor initiates a signaling cascade, which ultimately results in the activation of the Gli/Ci transcription factors. Suppressor of fused (Su(fu)) is a Gli/Ci-interacting protein, which acts as a negative regulator of Hh signaling in Drosophila and vertebrates. Su(fu) is also implicated as a tumor suppressor as its mutations have been found in medulloblastoma and prostate cancer. Su(fu) is thought to act by preventing the nuclear accumulation of Gli/Ci, however, mechanistic insight into its mode of action has remained elusive. We demonstrate here that Su(fu) prevents the nuclear accumulation of Gli1 and Gli2 through multiple mechanisms. While Su(fu) itself is not subject to CRM1-dependent regulation, Su(fu) sequesters Gli1 in the cytoplasm mostly through a mechanism that depends on the activity of the nuclear export protein CRM1. In contrast, CRM1-mediated export is not required for Su(fu) to sequester Gli2. Furthermore, we show that the N-terminus of Su(fu) is sufficient for Gli inactivation in the absence of cytoplasmic sequestration. Together, these observations reveal that Su(fu) regulates the activity of Gli1 and Gli2 through distinct cytoplasmic and nuclear mechanisms. PMID- 16316411 TI - Caspase-14 but not caspase-3 is processed during the development of fetal mouse epidermis. AB - The activation of caspases is a central step in apoptosis and may also be critical for terminal differentiation of epidermal keratinocytes (KC). In particular, caspase-3 has been implicated in the differentiation of embryonic KC as well as in programmed cell death of KC, and caspase-14 has been suggested to function in the formation or homeostasis of the stratum corneum (SC). To test the putative roles of these proteases, we determined their expression level and activation status during development of fetal mouse epidermis. The level of procaspase-3 did not change significantly during epidermal development, and enzyme activation was undetectable at any timepoint investigated. Despite the lack of active caspase-3, the newly formed stratum granulosum and the regressing periderm contained cells positive in the terminal deoxynucleotidyl transferase mediated fluorescein-dUTP nick end labeling assay, indicating that nuclear DNA was degraded without activation of caspase-3, thereby arguing against a proteolytic function of caspase-3 in embryonic KC differentiation. By contrast, caspase-14 increased in abundance from embryonic day 14.5 (E14.5) onwards and consistently localized to the suprabasal layers of fetal epidermis. The caspase 14 pro-enzyme was processed into its catalytic subunits, a step required for enzyme activity, on day E17.5, coinciding with SC formation. Thus, processing of procaspase-14 is not confined to air-exposed mature skin but also occurs during epidermal development in utero. In summary, this study demonstrates that caspase 14, but not caspase-3 activation coincides temporally and spatially with embryonic KC differentiation, suggesting a role for caspase-14 in terminally differentiated KC. PMID- 16316412 TI - Members of the lysyl oxidase family are expressed during the development of the frog Xenopus laevis. AB - Lysyl oxidase (Lox) is a copper-dependent amine oxidase that catalyzes the cross linking of collagen and elastin fibers in the extracellular matrix (ECM). In mammals, four closely related Lox-like enzymes have been described that share a highly conserved catalytic domain with Lox. We have characterized Xenopus laevis cDNAs for Lox, Loxl-1, and Loxl-3, and show that they are expressed during early embryonic development. Using RT-PCR we detected maternal transcripts for Xloxl-1, but levels remained low until tailbud stages. Transcripts for Xlox and Xloxl-3 were not detected until early neurulae, although transcripts for Xlox remained at low levels until tailbud stages. Whole mount in situ hybridization showed that transcripts for Xloxl-1 and Xloxl-3 are localized in the notochord, while transcripts for Xlox are found in the notochord, somites, and head. X. laevis Lox like enzymes were inhibited by incubating embryos, from cleavage stages to tadpole stages, in beta-aminopropionitrile, a specific inhibitor of the catalytic domain. The resulting embryos appeared to differentiate normally but suffered from poor collagen fiber formation. Defects included kinks in the notochord, a posterior shift of the somites, abnormal gut coiling, and the formation of edemas. Our data suggest that Lox-related enzymes are required for the proper formation of the ECM during X. laevis development. PMID- 16316413 TI - Lack of a laterality phenotype in Pkd1 knock-out embryos correlates with absence of polycystin-1 in nodal cilia. AB - The invariant asymmetric placement of thoracic and abdominal organs in the vertebrates is controlled by the left-asymmetric activity of the Nodal signaling cascade during embryogenesis. In the mouse embryo asymmetric induction of nodal is thought to be dependent on functional monocilia on the ventral node cells and on the Pkd2 gene, which encodes the calcium channel polycystin-2 (PC2). In humans mutations in PKD2 and PKD1 give rise to polycystic kidney disease. The PC1 and PC2 proteins are thought to function as part of a multifactorial complex. Localization of both proteins to the primary renal cilium suggested a function on cilia of the ventral node. Here we investigated Pkd1 knock-out embryos for laterality defects and found wild-type organ morphogenesis and normal expression of nodal and Pitx2. While PC2 localized to nodal cilia, no ciliary localization of PC1 was detected in mouse embryos. This finding was confirmed in an archetypical mammalian blastodisc, the rabbit embryo. Thus, absence of PC1 localization to cilia corresponded with a lack of laterality defects in Pkd1 knock-out embryos. Our results demonstrate a PC1-independent function of PC2 in left-right axis formation, and indirectly support a ciliary role of PC2 in this process. PMID- 16316415 TI - The Nobel prize in physiology or medicine 2005. PMID- 16316416 TI - Human CD56bright and CD56dim natural killer cell subsets respond differentially to direct stimulation with Mycobacterium bovis bacillus Calmette-Guerin. AB - Mycobacterium bovis bacillus Calmette-Guerin (BCG) is capable of directly stimulating several effector functions of human natural killer (NK) cells in the absence of interleukin-12 and professional antigen presenting cells. To assess the contribution of two main human NK-cell subsets (CD56(dim) and CD56(bright)) to the overall in vitro NK-cell response to BCG, peripheral blood mononuclear cells depleted of nylon wool-adherent cells or purified NK cells were stimulated with live BCG. By combining intranuclear bromodeoxyuridine (BrdU) staining and analysis of CD56 marker intensity, statistically higher percentages of BrdU(+) cells were found among the CD56(bright) subset than the CD56(dim) subset after 6 days of stimulation with BCG. Similarly, evaluation of intracellular interferon gamma (IFN-gamma) revealed that CD56(bright) cells were those mainly involved in IFN-gamma production in response to BCG. In contrast, the CD56(dim) subset contained higher levels of perforin and granzyme A, two key molecules for exocytosis-mediated cytotoxicity, than the CD56(bright) subset. Although 16-20-h stimulation with BCG did not substantially alter the expression of cytotoxic molecules by the two subsets, a decrease in perforin content was observed in the CD56(dim), but not in the CD56(bright) subset, following 4-h incubation with the NK-sensitive target K562 cell line. This decrease in perforin content correlated with the induction by BCG-stimulated NK cells, of early markers of apoptosis on target cells to a greater extent than unstimulated cells suggesting a major role for the CD56(dim) subset in cytotoxic activity in response to BCG. Taken together, these results demonstrate that CD56(bright) and CD56(dim) human NK-cell subsets exert different functional activities in response to a live bacterial pathogen. PMID- 16316417 TI - Detection of an autologous ligand for mannan-binding lectin on human B lymphocytes. AB - Mannan-binding lectin (MBL) is a collectin and a major soluble pattern recognition protein. MBL can distinguish self from nonself and altered self using its C-type carbohydrate recognition domain and may also interact via its collagen like region with autologous cells. Recently, it was found that MBL could bind to adherent cells (monocytes) and dendritic cells in a specific and sugar-sensitive manner. We have now investigated the interaction of MBL with fresh human peripheral blood cells and report binding to B lymphocytes and natural killer cells. The binding to B lymphocytes was studied in detail and was compared with the binding of MBL to monocytes and dendritic cells. Binding of MBL to B cells was evident at physiological MBL and calcium concentrations but was optimal at supraphysiological MBL concentrations. It was readily inhibited by autologous serum, mannan, mannose, GlcNAc and (to a lesser extent) galactose but not by C1q. A similar, but not identical, inhibition profile was observed with dendritic cells, but monocytes were not sensitive to mannose or mannan. We conclude that MBL is capable of binding to differently glycosylated ligands on several autologous cell types via its carbohydrate-recognition domain. We speculate that this could have functional significance at extravascular sites, but perhaps only in individuals possessing MBL genotypes conferring MBL sufficiency. PMID- 16316418 TI - The role of ERK5 in T-cell signalling. AB - The mitogen-activated protein kinase (MAPK) ERK5 plays an important role in mammary epithelial proliferation, endothelial cell survival and normal embryonic development. In nonhaematopoietic cells, mitogenic and stress signals activate the ERK5 cascade. Here, we investigated the role of the ERK5 pathway in T-cell activation and show that primary and leukaemic T cells express ERK5, whose activating phosphorylation is induced by antibodies against CD3 but not by phorbol myristate acetate treatment. ERK5 localized in the cytosol and nucleus in quiescent and activated T cells. In the latter, ERK5 phosphorylation was mainly observed in the nucleus. Selective activation of the ERK5 cascade by transfecting constitutively active MEK5 and wildtype ERK5 induced a reporter gene driven by the IL-2 promoter while barely affecting CD69 expression. These results suggest a new role for the ERK5 cascade in intracellular signalling in T cells. PMID- 16316419 TI - Analysis of epitopes on the unrelated proteins thyrotropin receptor and alpha 1 antitrypsin which are recognized by A10 monoclonal antibody. AB - In a previous study, we noticed an unexpected reaction of an antithyrotropin receptor ectodomain (ETSHR)-reactive monoclonal antibody, A10, with alpha1 antitrypsin (antitrypsin). Presently, we decided to probe the structural basis of this cross-reactivity. Recombinant ETSHR, antitrypsin, synthetic peptides corresponding to the region of similarity in these proteins (EEDFRV and EEDFHV, respectively) and a set of peptides related to this region, N- and C-terminally elongated, were used in the study. Comparing the values of the dissociation constants, we found that the affinity of peptides corresponding to the region of similarity to monoclonal antibody A10 was the same in spite of a difference in one residue (R 38 in ETSHR and H 209 in antitrypsin), whereas a change of E 206 to R in antitrypsin-related peptide dramatically decreased the affinity. The whole binding site of A10 in ETSHR as well as in antitrypsin was larger than the region of similarity. We propose that residues ECHQEEDFV represent the monoclonal antibody A10 epitope. They form an almost continuous sequence of residues 30-37 and 39 in ETSHR. The monoclonal antibody A10 binding site on antitrypsin is shorter. It comprises amino acids 205-208 and 210, from the region of similarity with, probably, additional two residues, H-287 and E 363. PMID- 16316420 TI - Mycoplasma mycoides ssp. mycoides biotype small colony-secreted components induce apoptotic cell death in bovine leucocytes. AB - Contagious bovine pleuropneumonia, caused by Mycoplasma mycoides ssp. mycoides biotype small colony (MmmSC), is one of the most serious cattle diseases in Africa. Several observations suggested that MmmSC had evolved an efficient way to escape the bovine immune responses by triggering host-cell cytotoxicity. This study was implemented to determine whether the cytotoxic effect was due to apoptotic cell death. To that end, bovine blood cells were cultured for up to 3 days in the presence of viable or heat-killed MmmSC compared to unstimulated cultures. The findings provided evidence for a viable MmmSC-induced, time dependent apoptosis in bovine blood leucocytes, whereas heat-killed MmmSC had no effect. Morphological and physiological changes (evidenced by TUNEL and annexin V staining) typical of apoptosis were observed in response to viable MmmSC. All the lymphocyte subsets as well as the monocyte/granulocyte subset exhibited extensive apoptosis after exposure to viable MmmSC. Our results demonstrated a potential role for MmmSC-secreted components as pathogenic factors able to induce programmed cell death in bovine blood leucocytes. PMID- 16316421 TI - Neutrophil activation in anti-proteinase 3-positive vasculitis--a prospective study. AB - In vitro studies have demonstrated that antineutrophil cytoplasmic antibodies (ANCA) have the capacity to activate neutrophils. Whether circulating neutrophils in patients with vasculitis are activated is under debate. Eight consecutive patients with antiproteinase 3 (PR3) positive acute vasculitis were included in this prospective study. Neutrophil expression of adhesion molecules, Fc-receptors and the ANCA-antigen PR3 was analysed and clinical characteristics were documented at inclusion and after 1, 3, 6 and 9 months in the same individuals. As additional markers of inflammation and endothelial activation interleukin-8 and soluble vascular cell adhesion molecule-1 in serum were analysed at the same time points. The expression of adhesion molecules on circulating neutrophils, CD62L and CD11b after in vitro N-formyl-methionyl-leucyl-phenylalanine stimulation was significantly decreased at diagnosis and after 1 month but returned to normal levels after 3-9 months. The neutrophil expression of Fc receptor IIIb (CD 16) was decreased at diagnosis but normalized after 1-9 months. The main finding was an activated neutrophil adhesion phenotype at diagnosis and after 1 month, with normalized expression of adhesion molecules at 3-9 months. A pathological regulation of adhesion molecules may have implications on the endothelial damage seen in vasculitis. PMID- 16316422 TI - Human primary adenotonsillar naive phenotype CD45RA CD4 T lymphocytes undergo apoptosis upon stimulation with a high concentration of CD3 antibody. AB - Young children need to develop immune tolerance to harmless foreign antigens such as digested nutrients and various inhaled airborne antigens. Because of its anatomical location, pharyngeal adenotonsillar tissue is a potential site for the establishment of this immune tolerance. To characterize possible mechanisms of peripheral immune tolerance, we studied human primary adenotonsillar naive phenotype CD45RA(+) CD4(+) T cells, which represent cells that have not previously encountered foreign antigens. It was found that these CD45RA(+) CD4(+) T cells expressed higher levels of the activation marker CD69 as compared with peripheral blood CD45RA(+) CD4(+) T cells. Upon stimulation with a high concentration of CD3 antibody, which mimics the encounter of a high antigen dose, adenotonsillar CD45RA(+) CD4(+) T lymphocytes, but not peripheral blood CD45RA(+) CD4(+) T cells, underwent apoptosis. After 6 h stimulation with a high concentration of CD3 antibody, over 25% of the cells were apoptotic. Interfering with the Fas-FasL interaction with recombinant Fas or an antibody against Fas ligand partially inhibited apoptosis. Our study results suggest that high concentrations of antigens, such as various nutrients and airborne antigens, may induce peripheral immune tolerance by selectively deleting naive phenotype CD45RA(+) CD4(+) T cells via T-cell receptor-triggered apoptosis in human adenotonsillar tissue. PMID- 16316423 TI - Elevated serum levels of soluble E-cadherin in patients with primary Sjogren's syndrome. AB - The aim of this study was to investigate serum levels of soluble E-cadherin (sE cadherin) in relation to lymphocytic organization and to characterize the expression of E-cadherin and integrin alphaEbeta7/CD103 in salivary gland epithelium of patients with Sjogren's syndrome (SS). Serum levels of sE-cadherin were significantly increased in SS compared to non-SS and nonsignificantly in germinal centre (GC)+ compared to GC- patients. Membrane-bound E-cadherin was detected on the majority of acinar and ductal epithelial cells in both SS and non SS. alphaEbeta7/CD103-positive cells were found scattered in focal infiltrates and GC, and in small clusters close to ductal and acinar epithelium at an increased level in SS compared to non-SS. Interestingly, E-cadherin-positive cells were detected randomly dispersed in focal lymphocytic infiltrates in 10/21 patients. By double-labelling, the cells with the E-cadherin-positive component were identified as CD68(+) macrophages. Elevated serum levels of sE-cadherin indicate an increased epithelial cell turnover and shedding, and sE-cadherin deserves further analysis as a potential diagnostic tool for SS. PMID- 16316424 TI - Influenza specific serum IgE is present in non-allergic subjects. PMID- 16316425 TI - Verbal intrusions precede memory decline in adults with Down syndrome. AB - BACKGROUND: Verbal intrusion errors are irrelevant responses made in the course of verbal memory retrieval or language production that have been associated with disruption of executive functions and the prefrontal cortex. They have been observed to occur more frequently both with normal aging and with neurodegenerative diseases such as Alzheimer's disease (AD). The purpose of this study was to longitudinally examine the production of verbal intrusions among middle-aged adults with Down syndrome (DS) and unspecified intellectual disability (ID) to determine whether producing verbal intrusions at one point in time was related to subsequent verbal memory performance. Because of the combination of a relative deficit in verbal working memory (WM), premature aging, and higher risk of AD among adults with DS, it was predicted that they would make more verbal intrusions than adults with unspecified ID. METHODS: Word List recall (WLR), the Selective Reminding Test (SRT), and the Cued Recall Test (CRT), were administered three times at 18-month intervals during a 3-year period. In Analysis 1, aetiology differences in making intrusion errors were examined. Twenty-three adults with unspecified ID in the moderate to mild range [time 1(T1) mean age = 47.2 years] and 42 adults with DS (T1 mean age = 44.3) participated. WLR is a serial WM task beginning at two word sequences and progressively increasing by one word every three trials. WLR intrusions were analysed because they were least likely to include 'educated guesses' because this test is not based on semantic categories. In Analysis 2, we only examined participants with DS. They were divided into two groups, 16 individuals who made at least one intrusion error at T1 (T1 mean age = 45.8) and 26 who did not (T1 mean age = 43.3). Longitudinal performance for these groups was analysed to determine whether the group that intruded at T1 did more poorly on subsequent memory tests. RESULTS: A higher proportion of responses comprised intrusions for the group with DS and a higher percentage of the participants with DS made at least one intrusion error when compared with participants with unspecified ID (74% and 44% respectively). Those participants with DS who made at least one intrusion error at T1 showed a subsequent decline in performance on both WLR and the SRT. CONCLUSIONS: The production of intrusion errors during a verbal WM task is a characteristic of middle-aged adults with DS. This suggests compromised executive function and control of inhibition within the verbal modality for this group. Further, verbal intrusions are a qualitative aspect of verbal processing that merit attention in considering the issue of deficiencies of language and verbal WM abilities among people with DS. Last, and perhaps most importantly, although not definitive diagnostically, an increase in verbal intrusions is a potentially noteworthy signal when evaluating the cognitive health of adults with DS. PMID- 16316426 TI - The outcomes of an intervention study to reduce the barriers experienced by people with intellectual disabilities accessing primary health care services. AB - BACKGROUND: People with intellectual disabilities (IDs) experience significant health inequalities compared with the general population. The barriers people with IDs experience in accessing services contribute to these health inequalities. Professionals' significant unmet training needs are an important barrier to people with IDs accessing appropriate services to meet their health needs. METHOD: A three group, pre- and post-intervention design was used to test the hypothesis that a training intervention for primary health care professionals would increase the knowledge and self-efficacy of participants. The intervention had two components - a written training pack and a 3-hour face-to-face training event. One group received the training pack and attended the training event, a second group received the training pack only, and a third group did not participate in the training intervention. Research measures were taken prior to the intervention and 3 months after the intervention. Statistical comparisons were made between the three groups. RESULTS: The participants in the training intervention reported that it had a positive impact upon their knowledge, skills and clinical practice. As a result of the intervention, 35 (81.4%) respondents agreed that they were more able to meet the needs of their clients with IDs, and 33 (66.6%) reported that they had made changes to their clinical practice. The research demonstrated that the intervention produced a statistically significant increase in the knowledge of participants (F = 5.6, P = 0.005), compared with the group that did not participate in the intervention. The self-efficacy of the participants that received both components of the intervention was significantly greater than the group that did not participate in the training (t = 2.079, P = 0.04). Participation in the two components of the training intervention was associated with significantly greater change in knowledge and self-efficacy than those receiving the training pack alone. CONCLUSION: This intervention was effective in addressing the measured training needs of primary health care professionals. Future research should directly evaluate the positive benefits of interventions on the lives of people with IDs. PMID- 16316427 TI - A measure of food seeking in individuals with Prader-Willi syndrome. AB - BACKGROUND: Individuals with Prader-Willi syndrome (PWS), a chromosome 15 genetic disorder, often have a significant preoccupation with food and problem behaviour related to food seeking is often prevalent. METHODS: In the present study, we compared how individuals with PWS responded on a survey regarding the acceptability of food in various locations that varied according to degree of appropriateness for human consumption (e.g. food on a plate, food in a garbage can). For a subgroup of participants, we observed how they actually responded when placed in a room with food items placed in the same locations depicted in the survey. In the first part of the study, three groups (25 typically developing individuals, 7 individuals with intellectual disability (ID), and 19 individuals with PWS) responded to a visual survey to determine the degree of acceptability of food items in various locations (e.g. on a table near a hairbrush, on the floor behind a toy box, in a trash can). In the second part of the study, these food items (popcorn, jelly beans) were placed in the 12 locations described above. Nine individuals diagnosed with PWS (deletion type) and three individuals with ID were given some break time in the room for 15 min. The amount of food consumed, the time spent food seeking, and time spent interacting with materials were measured. RESULTS: Results of the survey indicated that the PWS group differed significantly with regard to how they responded on the survey from the typically developing group, but did not differ significantly from the ID group. Results of the food seeking observations indicated that only three individuals with PWS ate a significant number of items. The three individuals did not differ from the rest of the group according to IQ or compulsivity score; however, they had significantly lower body mass index (BMI) scores and were younger than the other participants. CONCLUSIONS: The findings from the survey indicate that individuals with PWS are able to discriminate the appropriateness of eating items in more or less contaminated areas; however, the amount of time spent seeking food and the amount of food covertly consumed appeared to depend more directly on age and BMI. PMID- 16316428 TI - The general picture of supportive health environments for persons with intellectual disabilities among 121 disability welfare institutions in Taiwan. AB - BACKGROUND: Little information is available on the provision of supportive health environments for persons with intellectual disabilities (ID) in institutions. The aim of this study was to present an overview of supportive environments for health in institutions in Taiwan. METHODS: A cross-sectional survey was conducted to examine the perceptions of 121 Taiwanese Institutional Directors on their setting's implementation of supportive healthy physical, social, and economic environments. RESULTS: Analyses showed that first-aid kits (97.5%) and medicine cabinets (85.5%) were the most common health facilities in institutions. Seventy three per cent of institutions had set up specific areas to be used for rehabilitation practice, while only 43.1% thought their rehabilitation equipment/devices adequate for their real needs. Eighty-eight per cent of institutions implemented health promotion plans for people with ID, while 76.6% had appropriated specific health promotion plans. Sixty-three institutions (52.1%) reported employment of skilled nurses to serve people with ID, and these institutions showed statistically significant differences in implementation of each health facility. CONCLUSIONS: The present paper is the first to analyze supportive environments for health in disability institutions in Taiwan. An important focus of future research will be the extension of the present findings to consider the appropriateness of each area of supportive environments for improving the quality of institutional care for people with ID. PMID- 16316429 TI - The behavioural phenotype of Angelman syndrome. AB - BACKGROUND: The purpose of this review is to examine the notion of a behavioural phenotype for Angelman syndrome and identify methodological and conceptual influences on the accepted presentation. METHODS: Studies examining the behavioural characteristics associated with Angelman syndrome are reviewed and methodology is described. RESULTS: Potential bias in the description of the phenotype emerges with the use of case and cohort studies with the absence of comparison groups. A trend in the literature from a direct gene effect to a socially mediated effect on laughter is evident. CONCLUSION: Evidence for a behavioural phenotype of Angelman syndrome has begun to emerge. However, by adopting the concept of a 'behavioural phenotype', attention may become biased towards the underlying biological basis of the syndrome, with developmental and environmental factors being overlooked. PMID- 16316430 TI - Preferences in individuals with Angelman syndrome assessed by a modified Choice Assessment Scale. AB - BACKGROUND: Individuals with Angelman syndrome (AS) seem to have a strong preference for water-related items. Until present, preference assessment in AS has not been reported. METHODS: An adapted Dutch version of the Choice Assessment Scale (CAS) was administered by parents and other caregivers to 105 individuals with AS. The CAS was adapted by adding several items related to water and by adding a sub-scale describing activities and materials that individuals may avoid or escape. RESULTS: The five sub-scales and total scale of the modified CAS had good internal consistency. Water-related items were more often scored as preferred than non-water-related items. No associations were found between sub scale and total scale scores and demographic characteristics (e.g. genetic subtype, age). CONCLUSIONS: This study shows that people with AS often have strong preferences for water-related items. The modified CAS is a reliable and sensitive instrument to assess client preferences. PMID- 16316431 TI - The prevalence of intellectual disability in Limburg, the Netherlands. AB - BACKGROUND: Current changes in care philosophy and diversity in care arrangements caused the need for a new estimate of the number of people with intellectual disability (ID), based on recent data. Previous estimates were based on client registrations, which was thought unreliable at this time. This manuscript studies the question how many people with ID can be found in (part of) the Netherlands. METHODS: Identification of people with ID through a combination of general practice (GP) data bases and service registrations in the province of Limburg. RESULTS: The prevalence of people with ID appeared to be between 0.64% and 0.70%. About 0.21-0.27% were living with family or on their own; 55% of them did not use common ID services. CONCLUSION: Results are based on a combination of identification methods. Thirteen per cent of uncertain cases led to minimum and maximum estimates of the population. Limitations of the method and alternative ways of data collection are discussed. PMID- 16316432 TI - A long-term population-based clinical and morbidity review of Prader-Willi syndrome in Western Australia. AB - BACKGROUND: An investigation of the clinical morbidity and genetic profiles of individuals with Prader-Willi syndrome (PWS) in Western Australia (WA) was undertaken as part of a wider study into the effects of intellectual disability (ID) on the life course of individuals. METHODS: All persons with a diagnosis of PWS were identified from the records of the Disability Services Commission of WA (DSC). The DSC client files formed the main data source, and were supplemented by information from other state health data sets. The analysis was retrospective and quantitative in nature. RESULTS: A total of 56 individuals were identified, 10 of whom exhibited normal methylation patterns and so were analysed separately (PWS like). The ages of the PWS group ranged from 0.9 to 48.3 years, with six persons deceased. Most people with PWS (76%) had mild or moderate ID, and 70% lived in their family home. The birth prevalence of the disorder was 1 in 29 500 births. Respiratory disorders, dentistry and gastrointestinal disorders were common reasons for hospital admission, with epilepsy or convulsions also reported at moderate frequency. The PWS-like group shared many clinical features in common with PWS patients, the principal exceptions being hypotonia and feeding difficulties in infancy. CONCLUSIONS: The estimated birth prevalence of PWS was lower than expected; however, the case ascertainment method may have excluded some individuals. Older people with PWS were generally living in sheltered accommodation. As the cohort ages, demand for places in similar accommodation will increase, adding to the existing burden on service providers. Substantial future increases in the use of medical services and hospital-based care also are predicted with the onset of age-associated disorders. PMID- 16316433 TI - Development and evaluation of the CAHPS hospital survey. PMID- 16316434 TI - Measuring hospital care from the patients' perspective: an overview of the CAHPS Hospital Survey development process. AB - OBJECTIVE: To describe the developmental process for the CAHPS Hospital Survey. STUDY DESIGN: A pilot was conducted in three states with 19,720 hospital discharges. METHODS OF ANALYSIS: A rigorous, multi-step process was used to develop the CAHPS Hospital Survey. It included a public call for measures, multiple Federal Register notices soliciting public input, a review of the relevant literature, meetings with hospitals, consumers and survey vendors, cognitive interviews with consumer, a large-scale pilot test in three states and consumer testing and numerous small-scale field tests. FINDINGS: The current version of the CAHPS Hospital Survey has survey items in seven domains, two overall ratings of the hospital and five items used for adjusting for the mix of patients across hospitals and for analytical purposes. CONCLUSIONS: The CAHPS Hospital Survey is a core set of questions that can be administered as a stand alone questionnaire or combined with a broader set of hospital specific items. PMID- 16316435 TI - Review of the literature on survey instruments used to collect data on hospital patients' perceptions of care. AB - OBJECTIVE: To review the existing literature (1980-2003) on survey instruments used to collect data on patients' perceptions of hospital care. STUDY DESIGN: Eight literature databases were searched (PubMED, MEDLINE Pro, MEDSCAPE, MEDLINEplus, MDX Health, CINAHL, ERIC, and JSTOR). We undertook 51 searches with each of the eight databases, for a total of 408 searches. The abstracts for each of the identified publications were examined to determine their applicability for review. METHODS OF ANALYSIS: For each instrument used to collect information on patient perceptions of hospital care we provide descriptive information, instrument content, implementation characteristics, and psychometric performance characteristics. PRINCIPAL FINDINGS: The number of institutional settings and patients used in evaluating patient perceptions of hospital care varied greatly. The majority of survey instruments were administered by mail. Response rates varied widely from very low to relatively high. Most studies provided limited information on the psychometric properties of the instruments. CONCLUSIONS: Our review reveals a diversity of survey instruments used in assessing patient perceptions of hospital care. We conclude that it would be beneficial to use a standardized survey instrument, along with standardization of the sampling, administration protocol, and mode of administration. PMID- 16316436 TI - What do consumers want to know about the quality of care in hospitals? AB - OBJECTIVE: To guide the development of the Consumer Assessments of Healthcare Providers and Systems (CAHPS) Hospital Survey by identifying which domains of hospital quality included in a survey of recent hospital patients, and which survey items within those domains, would be of greatest interest to consumers and patients. DATA SOURCES/STUDY SETTING: Primary data were collected in four cities (Baltimore, Los Angeles, Phoenix, and Orlando), from a demographically varied mix of people of whom most, but not all, had recently been hospitalized or had a close loved one hospitalized. STUDY DESIGN/DATA COLLECTION METHOD: A total of 16 focus groups were held in these four cities. Groups were structured to be homogeneous with respect to type of health care coverage (Medicare, non Medicare), and type of recent hospital experience (urgent admission, elective admission, maternity admission, no admission). They were heterogeneous with respect to race/ethnicity, gender, and educational attainment. In addition to moderated discussions, focus group participants completed a pregroup questionnaire and various paper and pencil exercises during the groups. PRINCIPAL FINDINGS: A wide range of features were identified by participants as being relevant to hospital quality. Many were consonant with domains and items in the CAHPS Hospital Survey; however, some addressed structural features of hospitals and hospital outcomes that are not best derived from a patient experience survey. When shown the domains and items being considered for inclusion in the CAHPS Hospital Survey, participants were most interested in items relating to doctor communication with patients, nurse and hospital staff communication with patients, responsiveness to patient needs, and cleanliness of the hospital room and bathroom. Findings were quite consistent across groups regardless of location and participant characteristics. CONCLUSIONS: Consumers and patients have a high degree of interest in hospital quality and found a very high proportion of the items being considered for the CAHPS Hospital Survey to be so important they would consider changing hospitals in response to information about them. Hospital choice may well be constrained for patients, but publicly reported information from a patient perspective can also be used to support patient discussions with facilities and physicians about how to ensure patients have the best hospital experience possible. PMID- 16316437 TI - Role of cognitive testing in the development of the CAHPS Hospital Survey. AB - OBJECTIVE: To describe how cognitive testing results were used to inform the modification and selection of items for the Consumer Assessment of Health Providers and Systems (CAHPS) Hospital Survey pilot test instrument. DATA SOURCES: Cognitive interviews were conducted on 31 subjects in two rounds of testing: in December 2002-January 2003 and in February 2003. In both rounds, interviews were conducted in northern California, southern California, Massachusetts, and North Carolina. STUDY DESIGN: A common protocol served as the basis for cognitive testing activities in each round. This protocol was modified to enable testing of the items as interviewer-administered and self-administered items and to allow members of each of three research teams to use their preferred cognitive research tools. DATA COLLECTION/EXTRACTION METHODS: Each research team independently summarized, documented, and reported their findings. Item-specific and general issues were noted. The results were reviewed and discussed by senior staff from each research team after each round of testing, to inform the acceptance, modification, or elimination of candidate items. PRINCIPAL FINDINGS: Many candidate items required modification because respondents lacked the information required to answer them, respondents failed to understand them consistently, the items were not measuring the constructs they were intended to measure, the items were based on erroneous assumptions about what respondents wanted or experienced during their hospitalization, or the items were asking respondents to make distinctions that were too fine for them to make. Cognitive interviewing enabled the detection of these problems; an understanding of the etiology of the problem informed item revisions. However, for some constructs, the revisions proved to be inadequate. Accordingly, items could not be developed to provide acceptable measures of certain constructs such as shared decision making, coordination of care, and delays in the admissions process. CONCLUSIONS: Cognitive testing is the most direct way of finding out whether respondents understand questions consistently, have the information needed to answer the questions, and can use the response alternatives provided to describe their experiences or their opinions accurately. Many of the candidate questions failed to meet these standards. Cognitive testing only evaluates the way in which respondents understand and answer questions. Although it does not directly assess the validity of the answers, it is a reasonable premise that cognitive problems will seriously compromise validity and reliability. PMID- 16316438 TI - Methods used to streamline the CAHPS Hospital Survey. AB - OBJECTIVE: To identify a parsimonious subset of reliable, valid, and consumer salient items from 33 questions asking for patient reports about hospital care quality. DATA SOURCE: CAHPS Hospital Survey pilot data were collected during the summer of 2003 using mail and telephone from 19,720 patients who had been treated in 132 hospitals in three states and discharged from November 2002 to January 2003. METHODS: Standard psychometric methods were used to assess the reliability (internal consistency reliability and hospital-level reliability) and construct validity (exploratory and confirmatory factor analyses, strength of relationship to overall rating of hospital) of the 33 report items. The best subset of items from among the 33 was selected based on their statistical properties in conjunction with the importance assigned to each item by participants in 14 focus groups. PRINCIPAL FINDINGS: Confirmatory factor analysis (CFA) indicated that a subset of 16 questions proposed to measure seven aspects of hospital care (communication with nurses, communication with doctors, responsiveness to patient needs, physical environment, pain control, communication about medication, and discharge information) demonstrated excellent fit to the data. Scales in each of these areas had acceptable levels of reliability to discriminate among hospitals and internal consistency reliability estimates comparable with previously developed CAHPS instruments. CONCLUSION: Although half the length of the original, the shorter CAHPS hospital survey demonstrates promising measurement properties, identifies variations in care among hospitals, and deals with aspects of the hospital stay that are important to patients' evaluations of care quality. PMID- 16316439 TI - Exploratory factor analyses of the CAHPS Hospital Pilot Survey responses across and within medical, surgical, and obstetric services. AB - OBJECTIVES: To estimate the associations among hospital-level scores from the Consumer Assessments of Healthcare Providers and Systems (CAHPS) Hospital pilot survey within and across different services (surgery, obstetrics, medical), and to evaluate differences between hospital- and patient-level analyses. DATA SOURCE: CAHPS Hospital pilot survey data provided by the Centers for Medicare and Medicaid Services. STUDY DESIGN: Responses to 33 questionnaire items were analyzed using patient- and hospital-level exploratory factor analytic (EFA) methods to identify both a patient-level and hospital-level composite structures for the CAHPS Hospital survey. The latter EFA was corrected for patient-level sampling variability using a hierarchical model. We compared results of these analyses with each other and to separate EFAs conducted at the service level. To quantify the similarity of assessments across services, we compared correlations of different composites within the same service with those of the same composite across different services. DATA COLLECTION: Cross-sectional data were collected during the summer of 2003 via mail and telephone from 19,720 patients discharged from November 2002 through January 2003 from 132 hospitals in three states. PRINCIPAL FINDINGS: Six factors provided the best description of inter-item covariation at the patient level. Analyses that assessed variability across both services and hospitals suggested that three dimensions provide a parsimonious summary of inter-item covariation at the hospital level. Hospital-level factor structures also differed across services; as much variation in quality reports was explained by service as by composite. CONCLUSIONS: Variability of CAHPS scores across hospitals can be reported parsimoniously using a limited number of composites. There is at least as much distinct information in composite scores from different services as in different composite scores within each service. Because items cluster slightly differently in the different services, service specific composites may be more informative when comparing patients in a given service across hospitals. When studying individual-level variability, a more differentiated structure is probably more appropriate. PMID- 16316440 TI - Patterns of unit and item nonresponse in the CAHPS Hospital Survey. AB - OBJECTIVE: To examine the predictors of unit and item nonresponse, the magnitude of nonresponse bias, and the need for nonresponse weights in the Consumer Assessment of Health Care Providers and Systems (CAHPS) Hospital Survey. METHODS: A common set of 11 administrative variables (41 degrees of freedom) was used to predict unit nonresponse and the rate of item nonresponse in multivariate models. Descriptive statistics were used to examine the impact of nonresponse on CAHPS Hospital Survey ratings and reports. RESULTS: Unit nonresponse was highest for younger patients and patients other than non-Hispanic whites (p<.001); item nonresponse increased steadily with age (p<.001). Fourteen of 20 reports of ratings of care had significant (p<.05) but small negative correlations with nonresponse weights (median -0.06; maximum -0.09). Nonresponse weights do not improve overall precision below sample sizes of 300-1,000, and are unlikely to improve the precision of hospital comparisons. In some contexts, case-mix adjustment eliminates most observed nonresponse bias. CONCLUSIONS: Nonresponse weights should not be used for between-hospital comparisons of the CAHPS Hospital Survey, but may make small contributions to overall estimates or demographic comparisons, especially in the absence of case-mix adjustment. PMID- 16316441 TI - Equivalence of mail and telephone responses to the CAHPS Hospital Survey. AB - OBJECTIVE: To estimate the effect of survey mode (mail versus telephone) on reports and ratings of hospital care. DATA SOURCES/STUDY SETTING: The total sample included 20,826 patients discharged from a group of 24 distinct hospitals in three states (Arizona, Maryland, New York). We collected CAHPS data in 2003 by mail and telephone from 9,504 patients, of whom 39 percent responded by telephone and 61 percent by mail. STUDY DESIGN: We estimated mode effects in an observational design, using both propensity score blocking and (ordered) logistic regression on covariates. We used variables derived from administrative data (either included as covariates in the regression function or used in estimating the propensity score) grouped in three categories: individual characteristics, characteristics of the stay and hospital, and survey administration variables. DATA COLLECTION/EXTRACTION METHODS: We mailed a 66-item questionnaire to everyone in the sample and followed up by telephone with those who did not respond. PRINCIPAL FINDINGS: We found significant (p<.01) mode effects for 13 of the 21 questions examined in this study. The maximum magnitude of the survey mode effect was an 11 percentage-point difference in the probability of a "yes" response to one of the survey questions. Telephone respondents were more likely to rate care positively and health status negatively, compared with mail respondents. Standard regression-based case-mix adjustment captured much of the mode effects detected by propensity score techniques in this application. CONCLUSIONS: Telephone mode increases the propensity for more favorable evaluations of care for more than half of the items examined. This suggests that mode of administration should be standardized or carefully adjusted for. Alternatively, further item development may minimize the sensitivity of items to mode of data collection. PMID- 16316442 TI - Assessment of the equivalence of the Spanish and English versions of the CAHPS Hospital Survey on the quality of inpatient care. AB - OBJECTIVE: To describe translation and cultural adaptation procedures, and examine the degree of equivalence between the Spanish and English versions of the Agency for Healthcare Research and Quality's (AHRQ) Consumer Assessments of Healthcare Providers and Systems (CAHPS) Hospital Survey (H-CAHPS) of patient experiences with care. DATA SOURCES: Cognitive interviews on survey comprehension with 12 Spanish-speaking and 31 English-speaking subjects. Psychometric analyses of 586 responses to the Spanish version and 19,134 responses to the English version of the H-CAHPS survey tested in Arizona, Maryland, and New York in 2003. STUDY DESIGN: A forward/backward translation procedure followed by committee review and cognitive testing was used to ensure a translation that was both culturally and linguistically appropriate. Responses to the two language versions were compared to evaluate equivalence and assess the reliability and validity of both versions. DATA COLLECTION/EXTRACTION METHODS: Comparative analyses were carried out on the 32 items of the shortened survey version, focusing on 16 items that comprise seven composites representing different aspects of hospital care quality (communication with nurses, communication with doctors, communication about medicines, nursing services, discharge information, pain control, and physical environment); three items that rate the quality of the nursing staff, physician staff, and the hospital overall; one item on intention to recommend the hospital. The other 12 items used in the analyses addressed mainly respondent characteristics. Analyses included item descriptives, correlations, internal consistency reliability of composites, factor analysis, and regression analysis to examine construct validity. PRINCIPAL FINDINGS: Responses to both language versions exhibit similar patterns with respect to item-scale correlations, factor structure, content validity, and the association between each of the seven qualities of care composites with both the hospital rating and intention to recommend the hospital. Internal consistency reliability was slightly, yet significantly lower for the Spanish-language respondents for five of the seven composites, but overall the composites were generally equivalent across language versions. CONCLUSIONS: The results provide preliminary evidence of the equivalence between the Spanish and English versions of H-CAHPS. The translated Spanish version can be used to assess hospital quality of care for Spanish speakers, and compare results across these two language groups. PMID- 16316443 TI - Case-mix adjustment of the CAHPS Hospital Survey. AB - OBJECTIVES: To develop a model for case-mix adjustment of Consumer Assessment of Healthcare Providers and Systems (CAHPS) Hospital survey responses, and to assess the impact of adjustment on comparisons of hospital quality. DATA SOURCES: Survey of 19,720 patients discharged from 132 hospitals. METHODS: We analyzed CAHPS Hospital survey data to assess the extent to which patient characteristics predict patient ratings ("predictive power") and the heterogeneity of the characteristics across hospitals. We combined the measures to estimate the impact of each predictor ("impact factor") and selected high impact variables for adjusting ratings from the CAHPS Hospital survey. PRINCIPLE FINDINGS: The most important case-mix variables are: hospital service (surgery, obstetric, medical), age, race (non-Hispanic black), education, general health status (GHS), speaking Spanish at home, having a circulatory disorder, and interactions of each of these variables with service. Adjustment for GHS and education affected scores in each of the three services, while age and being non-Hispanic black had important impacts for those receiving surgery or medical services. Circulatory disorder, Spanish language, and Hispanic affected scores for those treated on surgery, obstetrics, and medical services, respectively. Of the 20 medical conditions we tested, only circulatory problems had an important impact within any of the services. Results were consistent for the overall ratings of nurse, doctor, and hospital. Although the overall impact of case-mix adjustment is modest, the rankings of some hospitals may be substantially affected. CONCLUSIONS: Case-mix adjustment has a small impact on hospital ratings, but can lead to important reductions in the bias in comparisons between hospitals. PMID- 16316447 TI - Asymmetric synthesis of chiral 4-substituted 5,5-diethyl oxazolidin-2-ones as potential effective chiral auxiliaries. AB - Four new chiral 4-substituted 5,5-diethyl oxazolidin-2-ones (5a-5d) as potential effective chiral auxiliaries were synthesized from readily available amino acids via asymmetric etherification, Boc protection, Grignard reaction and cyclization reactions in four steps with overall yields of 50-60%. PMID- 16316448 TI - Facile solid-phase synthesis of C-terminal peptide aldehydes and hydroxamates from a common Backbone Amide-Linked (BAL) intermediate. AB - C-Terminal peptide aldehydes and hydroxamates comprise two separate classes of effective inhibitors of a number of serine, aspartate, cysteine, and metalloproteases. Presented here is a method for preparation of both classes of peptide derivatives from the same resin-bound Weinreb amide precursor. Thus, 5 [(2 or 4)-formyl-3,5-dimethoxyphenoxy]butyramido-polyethylene glycol-polystyrene (BAL-PEG-PS) was treated with methoxylamine hydrochloride in the presence of sodium cyanoborohydride to provide a resin-bound methoxylamine, which was efficiently acylated by different Fmoc-amino acids upon bromo-tris-pyrrolidone phosphonium hexafluorophosphate (PyBrOP) activation. Solid-phase chain elongation gave backbone amide-linked (BAL) peptide Weinreb amides, which were cleaved either by trifluoroacetic acid (TFA) in the presence of scavengers to provide the corresponding peptide hydroxamates, or by lithium aluminum hydride in tetrahydrofuran (THF) to provide the corresponding C-terminal peptide aldehydes. With several model sequences, peptide hydroxamates were obtained in crude yields of 68-83% and initial purities of at least 85%, whereas peptide aldehydes were obtained in crude yields of 16-53% and initial purities in the range of 30-40%. Under the LiAlH4 cleavage conditions used, those model peptides containing t-Bu protected aspartate residues underwent partial side chain reduction to the corresponding homoserine-containing peptides. Similar results were obtained when working with high-load aminomethyl-polystyrene, suggesting that this chemistry will be generally applicable to a range of supporting materials. PMID- 16316449 TI - Improved solid-phase synthesis of alpha,alpha-dialkylated amino acid-rich peptides with antimicrobial activity. AB - A homologous series of nonapeptides and their acetylated versions were successfully prepared using solid-phase synthetic techniques. Each nonapeptide was rich in alpha,alpha-dialkylated amino acids [one 4-aminopiperidine-4 carboxylic acid (Api) and six alpha-aminoisobutyric acid (Aib) residues] and also included lysines or lysine analogs (two residues). The incorporation of the protected dipeptide 9-fluorenylmethyloxycarbonyl (Fmoc)-Aib-Aib-OH improved the purity and overall yields of these de novo designed peptides. The helix preference of each nonapeptide was investigated in six different solvent environments, and each peptide's antimicrobial activity and cytotoxicity were studied. The 3(10)-helical, amphipathic design of these peptides was born out most prominently in the N-terminally acetylated peptides. Most of the peptides exhibited modest activity against Escherichia coli and no activity against Staphylococcus aureus. The nonacetylated peptides (concentrations < or =100 microM) and the acetylated peptides (concentrations < or = 200 microM) did not exhibit any significant cytotoxicity with normal (nonactivated) murine macrophages. PMID- 16316450 TI - The effects of sucrose on stability of human brain natriuretic peptide [hBNP (1 32)] and human parathyroid hormone [hPTH (1-34)]. AB - Although the effect of sucrose on the physical stability of proteins has been well documented, its impact on their chemical stability is largely unknown. The aim of this study was to investigate the potential effects of sucrose on the structural conformation of human brain natriuretic peptide [hBNP (1-32)] and the synthetic human parathyroid hormone [hPTH (1-34)], and link these effects to chemical degradation pathways of these peptides. The stability of hBNP (1-32) and hPTH (1-34) was studied at pH 5.5. Aggregation was monitored using size exclusion high-performance liquid chromatography (SE-HPLC), whereas oxidation and deamidation products were measured by reversed phase (RP) HPLC. Fourier transform infrared (FT-IR) spectroscopy was used to study the peptides' conformation. Sucrose retarded aggregation, deamidation, and oxidation of hBNP (1-32) and hPTH (1-34), with a maximum effect at relatively high concentrations (as much as 1 m). FT-IR spectroscopy indicated that sucrose maintained the native conformation of hBNP (1-32) and induced small conformation changes in the hPTH (1-34) structure. Sucrose enhanced the stability of hBNP (1-32) and hPTH (1-34) in liquid formulations. The stabilizing effect of sucrose was due to a large extent to retardation of oxidation and deamidation of hBNP (1-32) and hPTH (1-34). PMID- 16316451 TI - Malignancy-induced autoimmunity to MUC1: initial antibody characterization. AB - Numerous reports document the existence of autoantibodies to MUC1 in the circulation of individuals with breast and other solid malignancies, with the majority of researchers utilizing MUC1 peptides in their detection. This report documents the purification, using peptide and whole molecule, and characterization of such autoantibodies from an individual with an unusual, highly MUC1-positive, myosarcoma. Purification of autoantibodies from serum was performed using affinity chromatography against either MUC1 peptide or whole molecule MUC1 [derived both from the patient (Pt-MUC1) and from a pool of sera from patients with advanced breast cancer (ABC-MUC1)]. Enzyme-linked immunosorbent assays (ELISAs) were used to compare specificity of purified autoantibodies. Peptide epitopes were determined by Ptifcan system against 7-mer peptides covering the 20 amino acid repeat of the MUC1 extracellular domain. Substantially higher amounts of autoantibodies were isolated when purifying against Pt-MUC1 rather than either ABC-MUC1 or peptide. Whole molecule purified autoantibodies demonstrated an increased specificity for tumour-derived MUC1. Pt MUC1 autoantibodies were of both the immunoglobulin (Ig)G and IgM class, whilst autoantibodies purified against ABC-MUC1 and MUC1 peptide were IgG only. A greater range of peptide epitopes was defined by those autoantibodies purified against whole molecule. This report presents data indicating the presence of autoantibodies to MUC1 in an individual diagnosed with a MUC1 over-expressing myosarcoma. Confirmation of these autoantibodies as being specific for tumour associated MUC1 is given. Further, it suggests that, although autoantibodies are present that recognize core protein determinants, the initial, and dominant, immunizing epitope is not purely pretentious in nature. PMID- 16316452 TI - The role of charged amphipathic helices in the structure and function of surfactant protein B. AB - Surfactant protein B (SP-B) is essential for normal lung surfactant function. Theoretical models predict that the disulfide cross-linked, N- and C-terminal domains of SP-B fold as charged amphipathic helices, and suggest that these adjacent helices participate in critical surfactant activities. This hypothesis is tested using a disulfide-linked construct (Mini-B) based on the primary sequences of the N- and C-terminal domains. Consistent with theoretical predictions of the full-length protein, both isotope-enhanced Fourier transform infrared (FTIR) spectroscopy and molecular modeling confirm the presence of charged amphipathic alpha-helices in Mini-B. Similar to that observed with native SP-B, Mini-B in model surfactant lipid mixtures exhibits marked in vitro activity, with spread films showing near-zero minimum surface tensions during cycling using captive bubble surfactometry. In vivo, Mini-B shows oxygenation and dynamic compliance that compare favorably with that of full-length SP-B. Mini-B variants (i.e. reduced disulfides or cationic residues replaced by uncharged residues) or Mini-B fragments (i.e. unlinked N- and C-terminal domains) produced greatly attenuated in vivo and in vitro surfactant properties. Hence, the combination of structure and charge for the amphipathic alpha-helical N- and C terminal domains are key to SP-B function. PMID- 16316453 TI - Secondary structure analysis of synthetic peptides of the extracellular domain of the human follicle-stimulating hormone receptor. AB - Follicle-stimulating hormone receptor (FSHR) is a glycoprotein hormone receptor and possesses a large extracellular domain (ECD) instrumental in hormone binding. The ECD is characterized by the presence of leucine-rich repeat (LRR) structures made up of alpha-helices flanked by beta-strands. Our previous studies with the synthetic peptides corresponding to the potentially surface-oriented regions of the ECD had led to the identification of some of these regions in either FSH binding or FSH-induced cAMP production or both. This study was undertaken with an aim to correlate the findings made in vitro with the secondary structures of the respective peptides. Accordingly, all peptides were screened for their secondary structures in different biochemical environments. This study correlates the observed alpha-helical signature with the previously demonstrated activity in signal generation for peptides 15-31 and 216-235 hFSHR, while FSH binding is correlated with the maintenance of beta-sheet structure in peptides 285-300 and 297-310 hFSHR as observed in vitro. PMID- 16316454 TI - Biological activities of cecropin B-thanatin hybrid peptides. AB - Ten kinds of hybrid peptides containing the N-terminal residues of cecropin B (CB) and C-terminal of thanatin (TH) were constructed and expressed as gluthathion S-transferase (GST)-fusion proteins. Variants were screened for the better biological activity, which was paralleled with the degree of growth inhibition of the transformant cells. The hybrid CB-TH g was selected as the best one among those hybrids by in vivo monitoring method and was chemical synthesized for in vitro antimicrobial activity analysis. The hybrid peptide showed rescued activity against several test strains when compared with the truncated isoforms of TH, suggesting that the peptides with different structure and mechanism could be used as templates for hybrid peptides design. PMID- 16316455 TI - Optimization of aromatic side chain size complementarity in the hydrophobic core of a designed coiled-coil. AB - The coiled-coil structure plays an important roles, especially in protein assembly. Previously we constructed AAB-type heterotrimeric coiled-coils by manipulating the packing in the hydrophobic core using Trp and Ala residues, where one Trp and two Ala residues were placed in the hydrophobic core instead of three Ile residues. To optimize the packing complementarity in the hydrophobic core, we investigated the effects of introducing various aromatic amino acids on the formation of an AAB-type heterotrimeric coiled-coil, by circular dichroism, thermal stability, and nuclear magnetic resonance (NMR) studies. We found that the Phe residue was more suitable for heterotrimeric coiled-coil formation than the Trp residue, when combined with two Ala residues, whereas the Tyr and His residues did not induce the coiled-coil structure efficiently. PMID- 16316456 TI - Structure-activity relationship of neuropeptide gamma derived from mammalian and fish. AB - This study of relationship between structure and biologic activity was performed using five neuropeptide gammas [NPgamma; mammalian-NPgamma (M-NPgamma), trout NPgamma (T-NPgamma), goldfish-NPgamma (G-NPgamma), bowfin-NPgamma (B-NPgamma), and shark-NPgamma (S-NPgamma)]. Circular dichroism (CD) spectra showed that all peptides took random structure in buffer solution. In neutral and acidic liposomes, M-NPgamma, T-NPgamma, B-NPgamma, and S-NPgamma still adopted random structure, while G-NPgamma had an alpha-helical structure. The biologic activity of NPgammas has been estimated by their effects on the intestinal motility and arterial relaxation. The intestinal motility was investigated with rat duodenum (RD), carp intestine (CI), and guinea-pig ileum (GPI). The arterial relaxing effect was tested with guinea-pig aorta (GPA) and rat mesenteric artery (RMA). In RD, the order of potency compared with the EC50 value was M-NPgamma >> S-NPgamma >> B-NPgamma >> G-NPgamma >> T-NPgamma. G-NPgamma was the most contractile agent in CI. S-NPgamma was the most contractile agent in GPI. Using an arterial relaxing test, the order of potency was G-NPgamma >> T-NPgamma >> B-NPgamma >> S NPgamma >> M-NPgamma in GPA, and all NPgammas remarkably reduced relaxing activity in RMA. Despite their structural similarities to NPgammas, G-NPgamma has high affinity to tachykinin receptor-binding sites in GPA and CI, indicating an alpha-helical structure may have a critical role for receptor binding. However, an alpha-helical structure does not play a critical role in recognizing receptor binding sites in RD and GPI. PMID- 16316457 TI - Synthesis and biologic activity of conformationally constrained analogs of L 363,301. AB - We report the synthesis, biological activity and conformational analysis of analogs of the cyclic hexapeptide L-363,301, c[Pro6-Phe7-D-Trp8-Lys9-Thr10-Phe11] (numbering as in the native hormone somatostatin-14). The d-Trp in position 8 was replaced with (2R,3S)- and (2R,3R)-beta-MeTrp respectively, with an added methyl group in the beta position of Trp. The objective of our study was to determine the potency and selectivity generated by the added constraint in the beta position of the d-Trp upon binding to human somatostatin receptors hsst1-5. We synthesized the building blocks enantioselectively and incorporated them into the peptides by SPPS. Competition binding assays revealed that both compounds 2 and 3 were selective for hsst2 over hsst5. The (2R,3S) analog 2 was approximately 30 times more potent at hsst2 than the (2R,3R) analog 3. Interestingly, the (2R,3R) compound showed no binding affinity at hsst5. PMID- 16316458 TI - How do caregivers know when to take their child for immunizations? AB - BACKGROUND: Childhood vaccinations help reduce and eliminate many causes of morbidity and mortality among children. The objective of this study was to compare 4:3:1:3:3 (4+ doses of diphtheria and tetanus toxoids and pertussis vaccine, 3+ doses of poliovirus vaccine, 1+ doses of measles-containing vaccine, 3+ doses of Haemophilus influenzae type b vaccine, and 3+ doses of hepatitis B vaccine) coverage among children whose caregivers learned by different methods when their child's most recent immunization was needed. METHODS: Between July 2001 and December 2002, a portion of households receiving the National Immunization Survey were asked how they knew when to take the child in for his/her most recent immunization. Responses were post-coded into several categories: 'Doctor/nurse reminder at previous immunization visit', 'Shot card/record', 'Reminder/recall', and 'Other'. Respondents could give more than one answer. Children who did not receive any vaccines, had < or = 1 visits for vaccinations, or whose caregiver did not provide an answer to the question were excluded from analyses. Chi-square analyses were used to compare 4:3:1:3:3 coverage among 19-35 month old children. RESULTS: Children whose caregivers indicated that a doctor/nurse told them at a previous immunization visit when to return for the next immunization had significantly greater 4:3:1:3:3 coverage than those who did not choose the response (77.2% vs. 70.1%, p < 0.01). However, no significant difference in coverage was found between households that did/did not indicate that reminder/recalls (71.0% vs. 75.5%, p = 0.24) helped them remember when to take their child for their most recent immunization visit; only borderline significance was found between those that did/did not choose shot cards (70.6% vs. 76.2%, p = 0.07). CONCLUSION: A doctor or nurse's reminder during an immunization visit of the next scheduled immunization visit effectively encourages caregivers to bring children in for immunizations, providing an inexpensive and easy way to effectively increase immunization coverage. PMID- 16316459 TI - First-line therapy with gemcitabine and paclitaxel in locally, recurrent or metastatic breast cancer: a phase II study. AB - BACKGROUND: This phase II study evaluated the efficacy and safety of gemcitabine (G) plus paclitaxel (T) as first-line therapy in recurrent or metastatic breast cancer. METHODS: Patients with locally, recurrent or metastatic breast cancer and no prior chemotherapy for metastatic disease received G 1200 mg/m2 on days 1 and 8, and T 175 mg/m2 on day 1 (before G) every 21 days for a maximum of 10 cycles. RESULTS: Forty patients, 39 metastatic breast cancer and 1 locally-advanced disease, were enrolled. Their median age was 61.5 years, and 85% had a World Health Organization performance status (PS) of 0 or 1. Poor prognostic factors at baseline included visceral involvement (87.5%) and > or =2 metastatic sites (70%). Also, 27 (67.5%) patients had prior adjuvant chemotherapy, 25 of which had prior anthracyclines. A total of 220 cycles (median 6; range, 1-10) were administered. Of the 40 enrolled patients, 2 had complete response and 12 partial response, for an overall response rate of 35.0% for intent-to-treat population. Among 35 patients evaluable for efficacy the response rate was 40%. Additional 14 patients had stable disease, and 7 had progressive disease. The median duration of response was 12 months; median time to progression, 7.2 months; median survival, 25.7 months. Common grade 3/4 toxicities were neutropenia in 17 (42.5%) patients each, grade 3 leukopenia in 19 (47.5%), and grade 3 alopecia in 30 (75.0%) patients; 1 (2.5%) patient had grade 4 thrombocytopenia. CONCLUSION: GT exhibited encouraging activity and tolerable toxicity as first-line therapy in metastatic breast cancer. Phase III trials for further evaluation are ongoing. PMID- 16316460 TI - A liver fibrosis cocktail? Psoriasis, methotrexate and genetic hemochromatosis. AB - BACKGROUND: Pathologists are often faced with the dilemma of whether to recommend continuation of methotrexate therapy for psoriasis within the context of an existing pro-fibrogenic risk factor, in this instance, patients with genetic hemochromatosis. CASE PRESENTATIONS: We describe our experience with two male psoriatic patients (A and B) on long term methotrexate therapy (cumulative dose A = 1.56 gms and B = 7.88 gms) with hetero- (A) and homozygous (B) genetic hemochromatosis. These patients liver function were monitored with routine biochemical profiling; apart from mild perivenular fibrosis in one patient (B), significant liver fibrosis was not identified in either patient with multiple interval percutaneous liver biopsies; in the latter instance this patient (B) had an additional risk factor of partiality to alcohol. CONCLUSION: We conclude that methotrexate therapy is relatively safe in patients with genetic hemochromatosis, with no other risk factor, but caution that the risk of fibrosis be monitored, preferably by non-invasive techniques, or by liver biopsy. PMID- 16316461 TI - The distance-profile representation and its application to detection of distantly related protein families. AB - BACKGROUND: Detecting homology between remotely related protein families is an important problem in computational biology since the biological properties of uncharacterized proteins can often be inferred from those of homologous proteins. Many existing approaches address this problem by measuring the similarity between proteins through sequence or structural alignment. However, these methods do not exploit collective aspects of the protein space and the computed scores are often noisy and frequently fail to recognize distantly related protein families. RESULTS: We describe an algorithm that improves over the state of the art in homology detection by utilizing global information on the proximity of entities in the protein space. Our method relies on a vectorial representation of proteins and protein families and uses structure-specific association measures between proteins and template structures to form a high-dimensional feature vector for each query protein. These vectors are then processed and transformed to sparse feature vectors that are treated as statistical fingerprints of the query proteins. The new representation induces a new metric between proteins measured by the statistical difference between their corresponding probability distributions. CONCLUSION: Using several performance measures we show that the new tool considerably improves the performance in recognizing distant homologies compared to existing approaches such as PSIBLAST and FUGUE. PMID- 16316462 TI - An algorithm for the determination and quantification of components of nucleic acid mixtures based on single sequencing reactions. AB - BACKGROUND: Determination and quantification of nucleic acid components in a mixture is usually accomplished by microarray approaches, where the mixtures are hybridized against specific probes. As an alternative, we propose here that a single sequencing reaction from a mixture of nucleic acids holds enough information to potentially distinguish the different components, provided it is known which components can occur in the mixture. RESULTS: We describe an algorithm that is based on a set of linear equations which can be solved when the sequencing profiles of the individual components are known and when the number of sequenced nucleotides is larger than the number of components in the mixture. We have implemented the procedure for one type of sequencing approach, pyrosequencing, which produces a stepwise output of peaks that is particularly suitable for the procedure. As an example we use signature sequences from ribosomal RNA to distinguish and quantify several different species in a mixture. Using simulations, we show that the procedure may also be applicable for dideoxy sequencing on capillary sequencers, requiring only some instrument specific adaptations of protocols and software. CONCLUSION: The parallel sequencing approach described here may become a simple and cheap alternative to microarray experiments which aim at routine re-determination and quantification of known nucleic acid components from environmental samples or tissue samples. PMID- 16316463 TI - Efficient single nucleotide polymorphism discovery in laboratory rat strains using wild rat-derived SNP candidates. AB - BACKGROUND: The laboratory rat (Rattus norvegicus) is an important model for studying many aspects of human health and disease. Detailed knowledge on genetic variation between strains is important from a biomedical, particularly pharmacogenetic point of view and useful for marker selection for genetic cloning and association studies. RESULTS: We show that Single Nucleotide Polymorphisms (SNPs) in commonly used rat strains are surprisingly well represented in wild rat isolates. Shotgun sequencing of 814 Kbp in one wild rat resulted in the identification of 485 SNPs as compared with the Brown Norway genome sequence. Genotyping 36 commonly used inbred rat strains showed that 84% of these alleles are also polymorphic in a representative set of laboratory rat strains. CONCLUSION: We postulate that shotgun sequencing in a wild rat sample and subsequent genotyping in multiple laboratory or domesticated strains rather than direct shotgun sequencing of multiple strains, could be the most efficient SNP discovery approach. For the rat, laboratory strains still harbor a large portion of the haplotypes present in wild isolates, suggesting a relatively recent common origin and supporting the idea that rat inbred strains, in contrast to mouse inbred strains, originate from a single species, R. norvegicus. PMID- 16316464 TI - Comparison of laboratory and immediate diagnosis of coagulation for patients under oral anticoagulation therapy before dental surgery. AB - BACKGROUND: Dental surgery can be carried out on patients under oral anticoagulation therapy by using haemostyptic measures. The aim of the study was a comparative analysis of coagulation by laboratory methods and immediate patient diagnosis on the day of the planned procedure. METHODS: On the planned day of treatment, diagnoses were carried out on 298 patients for Prothrombin Time (PT), the International Normalised Ratio (INR), and Partial Thromboplastin Time (PTT). The decision to proceed with treatment was made with an INR < 4.0 according to laboratory results. RESULTS: Planned treatment did not go ahead in 2.7% of cases. Postoperatively, 14.8% resulted in secondary bleeding, but were able to be treated as out-patients. 1.7% had to be treated as in-patients. The average error between the immediate diagnosis and the laboratory method: 95% confidence interval was -5.8 +/- 15.2% for PT, -2.7 +/- 17.9 s for PTT and 0.23 +/- 0.80 for INR. The limits for concordance were 9.4 and -21.1% for PT, 15.2 and -20.5 s for PTT, and 1.03 and -0.57 for INR. CONCLUSION: This study showed a clinically acceptable concordance between laboratory and immediate diagnosis for INR. Concordance for PT and PTT did not meet clinical requirements. For patients under oral anticoagulation therapy, patient INR diagnosis enabled optimisation of the treatment procedure when planning dental surgery. PMID- 16316465 TI - Development of antibodies to human embryonic stem cell antigens. AB - BACKGROUND: Using antibodies to specific protein antigens is the method of choice to assign and identify cell lineage through simultaneous analysis of surface molecules and intracellular markers. Embryonic stem cell research can be benefited from using antibodies specific to transcriptional factors/markers that contribute to the "stemness" phenotype or critical for cell lineage. RESULTS: In this report, we have developed and validated antibodies (either monoclonal or polyclonal) specific to human embryonic stem cell antigens and early differentiation transcriptional factors/markers that are critical for cell differentiation into definite lineage. CONCLUSION: These antibodies enable stem cell biologists to conveniently identify stem cell characteristics and to quantitatively assess differentiation. PMID- 16316466 TI - Astrocyte reactivity to Fas activation is attenuated in TIMP-1 deficient mice, an in vitro study. AB - BACKGROUND: Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a multifunctional secreted protein with pleiotropic actions, including the inhibition of matrix metalloproteinases (MMPs), cell death/survival and growth promoting activities. After inflammatory challenge, the levels of TIMP-1 are highly and selectively upregulated in astrocytes among glial cells, but little is know about its role in these neural cells. We investigated the influence of TIMP 1 null mutation in the reactivity of cultured astrocytes to pro-inflammatory stimuli with TNF-alpha and anti-Fas antibody. RESULTS: When compared to WT, mutant astrocytes displayed an overall increased constitutive gelatinase expression and were less responsive to Fas-mediated upregulation of MMP-9, of monocyte chemoattractant protein-1 (MCP-1) and of intercellular cell adhesion molecule-1 (ICAM-1), all markers of astrocyte inflammatory response. In contrast, TNF-alpha treatment induced all these factors similarly regardless of the astrocyte genotype. The incorporation of 3H-thymidin, a marker of cell proliferation, increased in wild-type (WT) astrocytes after treatment with anti Fas antibody or recombinant TIMP-1 but not in mutant astrocytes. Finally, lymphocyte chemotaxis was differentially regulated by TNF-alpha in WT and TIMP-1 deficient astrocytes. CONCLUSION: We provide evidence that the alteration of the MMP/TIMP balance in astrocytes influences their reactivity to pro-inflammatory stimuli and that Fas activation modulates the expression of members of the MMP/TIMP axis. We hypothesise that the Fas/FasL transduction pathway and the MMP/TIMP system interact in astrocytes to modulate their inflammatory response to environmental stimuli. PMID- 16316467 TI - Characterization of Toll-like receptors in primary lung epithelial cells: strong impact of the TLR3 ligand poly(I:C) on the regulation of Toll-like receptors, adaptor proteins and inflammatory response. AB - BACKGROUND: Bacterial and viral exacerbations play a crucial role in a variety of lung diseases including COPD or asthma. Since the lung epithelium is a major source of various inflammatory mediators that affect the immune response, we analyzed the inflammatory reaction of primary lung epithelial cells to different microbial molecules that are recognized by Toll-like receptors (TLR). METHODS: The effects of TLR ligands on primary small airway epithelial cells were analyzed in detail with respect to cytokine, chemokine and matrix metalloproteinase secretion. In addition, the regulation of the expression of TLRs and their adaptor proteins in small airway epithelial cells was investigated. RESULTS: Our data demonstrate that poly(I:C), a synthetic analog of viral dsRNA, mediated the strongest proinflammatory effects among the tested ligands, including an increased secretion of IL-6, IL-8, TNF-alpha, GM-CSF, GRO-alpha, TARC, MCP-1, MIP 3alpha, RANTES, IFN-beta, IP-10 and ITAC as well as an increased release of MMP 1, MMP-8, MMP-9, MMP-10 and MMP-13. Furthermore, our data show that poly(I:C) as well as type-1 and type-2 cytokines have a pronounced effect on the expression of TLRs and molecules involved in TLR signaling in small airway epithelial cells. Poly(I:C) induced an elevated expression of TLR1, TLR2 and TLR3 and increased the gene expression of the general TLR adaptor MyD88 and IRAK-2. Simultaneously, poly(I:C) decreased the expression of TLR5, TLR6 and TOLLIP. CONCLUSION: Poly(I:C), an analog of viral dsRNA and a TLR3 ligand, triggers a strong inflammatory response in small airway epithelial cells that is likely to contribute to viral exacerbations of pulmonary diseases like asthma or COPD. The pronounced effects of poly(I:C) on the expression of Toll-like receptors and molecules involved in TLR signaling is assumed to influence the immune response of the lung epithelium to viral and bacterial infections. Likewise, the regulation of TLR expression by type-1 and type-2 cytokines is important considering the impact of exogenous and endogenous TLR ligands on Th1 or Th2 driven pulmonary inflammations like COPD or asthma, respectively. PMID- 16316468 TI - Large mobile thrombus in non-atherosclerotic thoracic aorta as the source of peripheral arterial embolism. AB - The presence of thrombi in the atherosclerotic and/or aneurysmatic aorta with peripheral arterial embolism is a common scenario. Thrombus formation in a morphologically normal aorta, however, is a rare event. A 50 years old woman was admitted to the mergency department for pain, coldness, and anesthesia in the the left foot. She had a 25 years history of cigarette smoking, a history of postmenopausal hormone replacement therapy (HRT), hypercholesterolemia and hyperfibrinogenemia. An extensive serologic survey for hypercoagulability, including antiphospholipid antibodies, and vasculitis disorders was negative. Transesophageal echocardiography revealed a large, pedunculated and hypermobile thrombus attached to the aortic wall 5 cm distal of the left subclavian artery. The patient was admitted to the surgery department, where a 15 cm long fresh, parietal thrombus could be removed from the aorta showing no macroscopic wall lesions or any other morphologic abnormalities. This case report demonstrates the possibility of evolving a large, pedunculated thrombus in a morphologically intact aorta in a postmenopausal woman with thrombogenic conditions such as hyperfibrinogenemia, hypercholesterolemia, smoking and HRT. For these patients, profiling the individual risk and weighing the benefits against the potential risks is warranted before prescribing HRT. PMID- 16316469 TI - Pharmacological reversal of endothelin-1 mediated constriction of the spiral modiolar artery: a potential new treatment for sudden sensorineural hearing loss. AB - BACKGROUND: Vasospasm of the spiral modiolar artery (SMA) may cause ischemic stroke of the inner ear. Endothelin-1 (ET-1) induces a strong, long-lasting constriction of the SMA by increasing contractile apparatus Ca2+ sensitivity via Rho-kinase. We therefore tested several Rho-kinase inhibitors and a cell permeable analogue of cAMP (dbcAMP) for their ability to reverse ET-1-induced constriction and Ca2+-sensitization. METHODS: The present study employed SMA isolated from gerbil temporal bones. Ca2+sensitivity was evaluated by correlating vascular diameter and smooth muscle cell [Ca2+]i, measured by fluo-4 microfluorometry and videomicroscopy. RESULTS: The Rho-kinase inhibitors Y-27632, fasudil, and hydroxy-fasudil reversed ET-1-induced vasoconstriction with an IC50 of 3, 15, and 111 micromol/L, respectively. DbcAMP stimulated a dose-dependent vasodilation (Ec50 = 1 mmol/L) and a reduction of [Ca2+]i (EC50 = 0.3 micromol/L) of ET-1-preconstricted vessels (1 nmol/L). Fasudil and dbcAMP both reversed the ET-1-induced increase in Ca2+ sensitivity. CONCLUSION: Rho-kinase inhibition and dbcAMP reversed ET-1-induced vasoconstriction and Ca2+-sensitization. Therefore, Rho-kinase inhibitors or cAMP modulators could possess promise as pharmacological tools for the treatment of ET-1-induced constriction, ischemic stroke and sudden hearing loss. PMID- 16316470 TI - Melatonin, immune function and aging. AB - Aging is associated with a decline in immune function (immunosenescence), a situation known to correlate with increased incidence of cancer, infectious and degenerative diseases. Innate, cellular and humoral immunity all exhibit increased deterioration with age. A decrease in functional competence of individual natural killer (NK) cells is found with advancing age. Macrophages and granulocytes show functional decline in aging as evidenced by their diminished phagocytic activity and impairment of superoxide generation. There is also marked shift in cytokine profile as age advances, e.g., CD3+ and CD4+ cells decline in number whereas CD8+ cells increase in elderly individuals. A decline in organ specific antibodies occurs causing reduced humoral responsiveness. Circulating melatonin decreases with age and in recent years much interest has been focused on its immunomodulatory effect. Melatonin stimulates the production of progenitor cells for granulocytes-macrophages. It also stimulates the production of NK cells and CD4+ cells and inhibits CD8+ cells. The production and release of various cytokines from NK cells and T-helper lymphocytes also are enhanced by melatonin. Melatonin presumably regulates immune function by acting on the immune-opioid network, by affecting G protein-cAMP signal pathway and by regulating intracellular glutathione levels. Melatonin has the potential therapeutic value to enhance immune function in aged individuals and in patients in an immunocompromised state. PMID- 16316471 TI - A cross-sectional study of the association between persistent organochlorine pollutants and diabetes. AB - BACKGROUND: Experimental evidence supports the hypothesis that persistent organochlorine pollutants (POPs) may cause type 2 diabetes mellitus, whereas there is no fully convincing epidemiological evidence for such an association. In Sweden the most important source of POP exposure is fatty fish. We have assessed the association between serum levels of POPs and prevalence of diabetes in Swedish fishermen and their wives, with high consumption of fatty fish from the Baltic Sea. METHODS: In 196 men (median age 60 years) and 184 women (median age 64 years), we analyzed 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153) and 1,1 dichloro-2,2-bis(p-chlorophenyl)-ethylene (p,p'-DDE) in serum using gas chromatography-mass spectrometry. The participants were asked if they had diabetes and, if so, since which year and about medication and diet. The Odds Ratios (OR) for diabetes with respect to continuous exposure variables were analyzed with logistic regression, adjusting for potential confounders. Moreover trends of diabetes prevalence with respect to trichotomized exposure variables were tested with Jonckheere-Terpstra's test. RESULTS: Six percent of the men and 5% of the women had diabetes. After confounder adjustment CB-153 was significantly associated with diabetes prevalence using both categorized and continuous exposure data (an increase of 100 ng/g lipid corresponded to an OR of 1.16, 95% confidence interval [CI] 1.03, 1.32, p = 0.03). Similar associations were observed for p,p'-DDE (an increase of 100 ng/g lipid corresponded to an OR of 1.05, 95% CI 1.01, 1.09, p = 0.006). Gender stratified analyses showed among men consistent positive associations with CB-153, but a more ambiguous pattern with respect to DDE. In contrast, among the women the associations with p,p'-DDE were stronger than with CB-153. CONCLUSION: The study provides support that POP exposure might contribute to type 2 diabetes mellitus. PMID- 16316472 TI - Long-term use of non-steroidal anti-inflammatory drugs and the risk of myocardial infarction in the general population. AB - BACKGROUND: Recent data indicate that chronic use of coxibs leads to an increased occurrence of thrombotic cardiovascular events. This raises the question as to whether traditional non-steroidal anti-inflammatory drugs (tNSAIDs) might also produce similar hazards. Our aim has been to evaluate the association between the chronic use of tNSAIDs and the risk of myocardial infarction (MI) in patients. METHODS: We performed a nested case-control analysis with 4,975 cases of acute MI and 20,000 controls, frequency matched to cases by age, sex, and calendar year. RESULTS: Overall, current use of tNSAID was not associated with an increased risk of MI (RR:1.07;95%CI: 0.95-1.21). However, we found that the relative risk (RR) of MI for durations of tNSAID treatment of >1 year was 1.21 (95% CI, 1.00-1.48). The corresponding RR was 1.34 (95% CI, 1.06-1.70) for non-fatal MI. The effect was independent from dose. The small risk associated with long-term use of tNSAIDs was observed among patients not taking low-dose aspirin (RR: 1.29; 95% CI, 1.01-1.65). The effect of long-term use for individual tNSAIDs ranged from a RR of 0.87 (95% CI, 0.47-1.62) with naproxen to 1.38 (95% CI, 1.00-1.90) with diclofenac. CONCLUSION: This study adds support to the hypothesis that chronic treatment with some tNSAIDs is associated with a small increased risk of non fatal MI. Our data are consistent with a substantial variability in cardiovascular risks between individual tNSAIDs. PMID- 16316473 TI - Primary care use of antipsychotic drugs: an audit and intervention study. AB - BACKGROUND: Concerns regarding the use of antipsychotic medication in secondary care suggested an examination of primary care prescribing. AIM: To audit and intervene in the suboptimal prescribing of antipsychotic drugs to primary care patients. DESIGN OF STUDY: Cross-sectional prevalence: subsequent open treatment intervention. SETTING: Seven of the 29 practices in the Eastern Hull Primary Care Trust. METHODS: Criteria for best practice were developed, against which prescribing standards were tested via audit. Patients identified as suboptimally prescribed for were invited to attend an expert review for intervention. RESULTS: 1 in 100 of 53,000 patients was prescribed antipsychotic treatment. Diagnoses indicating this were impossible to ascertain reliably. Half the regimes failed one or more audit criteria, leaving diagnosis aside. Few practices agreed to patients being approached: of 179 invitations sent, only 40 patients attended. Of 32 still taking an antipsychotic drug, 26 required changes. Mean audit criteria failed were 3.4, lack of psychotic disorder diagnosis and problematic side effects being most frequent. Changes were fully implemented in only 16 patients: reasons for complete or partial failure to implement recommendations included the wishes or inaction of patients and professionals, and worsening of symptoms including two cases of antipsychotic withdrawal syndrome. CONCLUSION: Primary care prescribing of antipsychotic drugs is infrequent, but most is unsatisfactory. Intervention is hampered by pluralistic reluctance: even with expert guidance, rationalisation is not without risk. Use of antipsychotic drugs in primary care patients whose diagnosis does not warrant this should be avoided. HOW THIS FITS IN: This study adds to concerns regarding high levels of off licence use of potentially harmful medication. It adds evidence of major difficulties in rationalizing suboptimal regimes despite expert input. Relevance to the clinician is that it is better to avoid such regimes in the first place especially if there is no clear 'exit strategy': if in doubt, seek a specialist opinion. PMID- 16316474 TI - Outcome following emergency surgery for refractory severe ulcerative colitis in a tertiary care centre in India. AB - BACKGROUND: Steroid-based intensive medical therapy for severe ulcerative colitis is successful in 60-70% of such patients. Patients with complications or those refractory to medical therapy require emergency colectomy for salvage. Little is known about the impact of timing of surgical intervention and surgical outcomes of such patients undergoing emergency surgery in India where the diagnosis is often delayed or missed in patients who are poor, malnourished and non-compliant to medical treatment. METHODS: The clinical records of all patients undergoing emergency surgery for severe ulcerative colitis or its complication in the Department of GI surgery AIIMS, New Delhi, India, between January 1985 and December 2003 were retrieved and data pertaining to demographic features, duration of intensive medical therapy, presence of complications, time from admission to emergency surgery, surgical procedure, in-hospital morbidity and mortality and follow up status extracted. RESULTS: A total of 72 patients underwent emergency surgery (Subtotal colectomy: 60; ileostomy alone under local anaesthesia: 12). Poor nutritional status was seen in 61% of the patients. Twenty one patients (29%) underwent emergency surgery for complications of severe ulcerative colitis such as colonic perforation (spontaneous 6, iatrogenic 4), massive lower gastrointestinal haemorrhage (5), toxic megacolon (4) and large bowel obstruction (2). The remaining patients (n = 51) underwent emergency surgery following failed intensive therapy; 17 underwent surgery < or = 5 days (Group I) and 34 were operated > 5 days (Group II) after initiation of intensive therapy. In this group all the post-operative deaths (n = 8) occurred in those who were operated after 5 days. The difference in mortality in these two groups (i.e. surgical intervention < or = or > 5 days) was statistically significant {0/17 (Group I) vs 8/34 (Group II); p = 0.03}. Overall, 12 patients died (in hospital mortality: 16.7%). The mortality was higher (10/43; 23.3%) in our early experience (i.e. 1985-1995) when compared to our subsequent experience (2/29; 6.9%) (1996-2003). A total of 48 patients (including 3 awaiting a restorative procedure) are alive on follow up (66.7%; 3 patients lost to follow up). A restorative procedure could be successfully completed in 81% of the survivors of the emergency procedure. CONCLUSION: To optimize the outcome, a combined team of physicians and surgeons should be involved in the management of patients with severe ulcerative colitis with focus on nutritional support, correction of metabolic derangements, close clinical monitoring and timely assessment for the need for emergency surgery. This retrospective analysis shows that improved results can be achieved with experience and by following a policy of early surgical intervention within 5 days, especially in patients who have failed intensive medical therapy. PMID- 16316476 TI - Association of the functional -1019C/G 5-HT1A polymorphism with prefrontal cortex and amygdala activation measured with 3 T fMRI in panic disorder. AB - Serotonergic genes have been implicated in the pathogenesis of panic disorder and amygdala function in response to fearful stimuli. Regional brain activation on visual presentation of emotional facial stimuli was investigated in 20 patients with panic disorder by means of fMRI at 3 T. All patients were genotyped for the functional -1019C/G 5-HT1A and 5-HTTLPR polymorphisms. In patients homozygous for the 5-HT1A -1019G risk allele (n=5), fearful stimuli were associated with a decreased activation of right prefrontal cortex regions. Patients homozygous for the 5-HT1A -1019G risk allele or patients carrying the short risk allele of the 5 HTTLPR (n=13) showed higher amygdala activation in response to happy faces. This exploratory study suggests a role of the functional -1019C/G 5-HT1A and 5-HTTLPR polymorphisms on prefrontal cortex and amygdala activation patterns in response to emotional facial stimuli. These serotonergic polymorphisms might increase the risk for panic disorder by contributing to an altered processing of emotional stimuli. PMID- 16316477 TI - Magnetoencephalography (MEG): a non-invasive tool for studying cortical effects in psychopharmacology. AB - Magnetoencephalography (MEG) is a non-invasive method for studying magnetic fields generated by simultaneously firing neurons outside the skull. The skull, scalp and brain tissue do not distort magnetic fields, so the cortical activity can be easily measured. MEG is starting to be used to explore the effects of various psychopharmacological agents on resting brain, sensory and cognitive processing. Scopolamine and agents enhancing GABA functions have shown differential effects on cortical neural oscillations. Further, with GABA, serotonin, dopamine and acetylcholine transmissions have differential effects on early cortical and pre-attentional processing in the auditory and frontal cortices. Monoamines also differently regulate the activity of the somatosensory cortex. Taken together, MEG with a resolution of milliseconds allows exploration of focal cortical effects of psychopharmacological agents giving information different from other brain-imaging modalities. PMID- 16316478 TI - A meta-analysis of profile and time-course of symptom change in acute schizophrenia treated with atypical antipsychotics. AB - The profile and time-course of symptom response in acute schizophrenia is unclear. For the present study, we hypothesized that the time-course would be nonlinear. A meta-analysis was performed using randomized, controlled clinical trials of five atypical antipsychotics reported in nine electronic databases. Studies were of subjects experiencing an acute exacerbation of illness, with multiple BPRS or PANSS data-points as outcome measures. A mixed factorial repeated-measures ANOVA was used. Twenty-one published clinical trials were identified. Reduction in total symptoms from baseline to 4 wk was associated with a linear decline in symptomatology (F=23.4, d.f.=1, 7, p=0.002) without attenuation of effect. In contrast, from baseline to 6 wk the linear symptom reduction (F=76.5, d.f.=1, 12, p<0.001) eventually flattened at the end of the trial (F=87.2, d.f.=1, 12, p<0.001). Secondary analyses showed a similar pattern for typical antipsychotics, and the same profile for risperidone and olanzapine as for atypical agents as a whole. Inclusion of LOCF data altered the results at 4 wk, but not 6 wk; completion rates had no effect on results. In conclusion, this meta-analysis confirms our hypothesis for 6-wk data. The profile of symptom change is one of linear symptom reduction until 4 wk, with a flattening of treatment effects by 6 wk. A curvilinear profile of schizophrenia symptom reduction has possible implications with respect to trial design and clinical decision-making. PMID- 16316479 TI - Nicotine blocks stress-induced impairment of spatial memory and long-term potentiation of the hippocampal CA1 region. AB - The effect of chronic nicotine treatment on chronic psychosocial stress-induced impairment of short-term memory and long-term potentiation (LTP) was determined. An "intruder" stress model was used to induce psychosocial stress for 4-6 wk, during which rats were injected with saline or nicotine (1 mg/kg s.c.) twice a day. The radial arm water maze memory task was used to test hippocampus-dependent spatial memory. Chronic psychosocial stress impaired short-term memory without affecting the learning phase or long-term memory. Concurrent chronic nicotine treatment prevented stress-induced short-term memory impairment. In normal rats chronic nicotine treatment had no effect on learning and memory. Extracellular recordings from the CA1 region of anaesthetized rats showed severe reduction of LTP magnitude in stressed rats, which was normalized in nicotine-treated stressed rats. Nicotine had no effect on LTP in control animals. These results showed that chronic nicotine treatment improved hippocampus-dependent spatial memory and LTP only when impaired by stress. PMID- 16316480 TI - Secular trends in first hospitalizations for major mood disorders with comorbid substance use. AB - In the past half-century, the incidence of major mood disorders reportedly has risen, and onset age, diminished. Substance-use disorders (SUDs) are commonly comorbid with mood disorders and may influence their course and outcome. Since secular relationships of these disorders remain unknown, we evaluated medical records of 421 patients (233 men, 188 women) at first-lifetime admissions for major depression, mania or mixed bipolar episodes at a public hospital in Italy during 1978-2002, updated diagnoses to DSM-IV criteria, and compared selected factors between subjects with vs. without SUDs, seeking evidence of secular trends in SUDs in association with early mood disorders. SUD was diagnosed in 122 patients (29.0%). SUD risk was greatest in young males. Relative risk (RR) of diagnostic association with SUDs ranked: mixed states (RR 1.80), mania (RR 1.06), depression (RR 0.55). Annual rates of comorbid SUD and mood disorders increased continuously over the 25-yr epoch for all substances (r=0.640, p<0.001). Age at onset of illness and at first hospitalization (averaging 36 yr and 38 yr) were unrelated to year of hospitalization in this relatively brief sampling, but patients with SUD were younger at onset and admission, overall. Clinical Global Impression (CGI) ratings of illness severity at hospitalization and discharge were stable across years. These findings indicate a secular increase of comorbid SUD among first admissions for mood disorder, especially in young males, with a parallel increase in the proportion of bipolar disorder diagnoses over the past quarter of a century. PMID- 16316481 TI - Exposure to amphetamine in rats during periadolescence establishes behavioural and extrastriatal neural sensitization in adulthood. AB - Drug abuse during adolescence may predispose towards later adult substance abuse and major depressive disorders (Brook et al., 2002). The purpose of the present study was to characterize whether behavioural sensitization to amphetamine occurred in adult rats following adolescent exposure to amphetamine [low (2 mg/kg.d) or high (10 mg/kg.d) i.p. for 10 d] and the pattern of neural activation associated with sensitized behaviour, in male Sprague-Dawley rats. Following initial treatment (post-natal days 33-41) and a subsequent 4-wk period of abstinence, rats initially treated with either amphetamine regime showed a similar sensitized locomotor activity upon re-challenge with amphetamine (1.5 mg/kg i.p.) compared to rats acutely challenged with this dose of amphetamine. Fos-IR expression in the "high" sensitized group was significantly greater than acutely challenged rats in all quadrants of the CPu. Both "low" and "high" sensitized groups demonstrated heightened Fos expression relative to acutely challenged rats in the medial and central amygdala, as well as rostroventrolateral medulla, whereas Fos-IR in the locus coeruleus and substantia nigra pars reticulata was significantly increased only in the "high" sensitized group compared to acute. Double-labelling for tyrosine hydroxylase confirmed an absence of Fos-IR in A9 and A10 regions. The present study has shown a robust and persistent sensitization in adulthood to amphetamine re-challenge following initial adolescent exposure in rats. This manifestation of sensitization apparently results in widespread neural activation in limbic and autonomic structures. PMID- 16316482 TI - Prevention of relapse in generalized anxiety disorder by escitalopram treatment. AB - Escitalopram has demonstrated a robust and dose-dependent efficacy in the treatment of generalized anxiety disorder (GAD) for up to 3 months. In the present study, the efficacy and tolerability of escitalopram in the prevention of relapse in GAD was investigated. A total of 491 patients with a primary diagnosis of GAD and a Hamilton Anxiety (HAMA) total score>or=20 received 12 wk of open label treatment with a fixed dose of escitalopram (20 mg/d). Of these, 375 patients responded (HAMA total scoreor=15, or lack of efficacy, as judged by the investigator. The results of the primary analysis showed a clear beneficial effect of escitalopram relative to placebo on the time to relapse of GAD (log-rank test, p<0.001). The risk of relapse was 4.04 times higher for placebo-treated patients than for escitalopram-treated patients; the proportion of patients who relapsed was statistically significantly higher in the placebo group (56%) than in the escitalopram group (19%) (p<0.001). Escitalopram was well tolerated and 7% of the escitalopram-treated patients withdrew due to adverse events, vs. 8% of the placebo patients. The incidence of discontinuation symptoms with escitalopram during tapered withdrawal was low; the symptoms primarily being dizziness (10-12%), nervousness (2-6%), and insomnia (2 6%). Escitalopram 20 mg/d significantly reduced the risk of relapse and was well tolerated in patients with GAD. PMID- 16316483 TI - Selective effects of typical antipsychotic drugs on SNAP-25 and synaptophysin in the hippocampal trisynaptic pathway. AB - Recent studies indicate that levels of presynaptic proteins are altered in the post-mortem brain in schizophrenia. In particular, the hippocampus exhibits reduced levels of synaptophysin and the SNARE protein SNAP-25. The effects of treatment with antipsychotic drugs on levels of SNAP-25 in the hippocampus remains unknown. To determine the effects of typical antipsychotic drugs on levels of synaptophysin and SNAP-25 in the hippocampus, rats were treated with chlorpromazine, haloperidol or trifluoperazine for 21 d. Quantitative immunohistochemistry was used to measure immunoreactivity within the trisynaptic circuit of the hippocampus. Trifluoperazine decreased synaptophysin within the Schaffer collateral region of the radiatum lacunosum in CA1, while haloperidol and chlorpromazine increased SNAP-25 throughout the trisynaptic pathway of the hippocampus, with strongest effects in the mossy fibre region of CA3. These results indicate that presynaptic proteins represent a potential molecular substrate for the effects of antipsychotic drugs on hippocampal synaptic connectivity. PMID- 16316484 TI - Facial expression perception: an objective outcome measure for treatment studies in mood disorders? AB - Facial expressions are important cues used in social communication. Studies in both patients with mood disorders and healthy volunteers have shown that facial expression perception can vary according to current mood state. Interpretation or perception of facial expressions can also be altered by administration of certain psychopharmacological agents. Novel drug development at present is restricted by the lack of valid and sensitive markers of mood state. This review suggests that measurement of facial expression perception may prove to be a useful experimental tool for assessing efficacy of antidepressant treatments. PMID- 16316485 TI - Treatment of behavioural, cognitive and circadian rest-activity cycle disturbances in Alzheimer's disease: haloperidol vs. quetiapine. AB - This 5-wk, open-label, comparative study investigated the effects of quetiapine and haloperidol on behavioural, cognitive and circadian rest-activity cycle disturbances in patients with Alzheimer's disease (AD). Out of a total of 30 patients enrolled in the study, there were 22 completers, 11 in the quetiapine group (mean age 81.9+/-1.8 yr, mean baseline MMSE 19.9+/-1.3, mean dose 125 mg) and 11 in the haloperidol group (mean age 82.3+/-2.5 yr, mean baseline MMSE 18.1+/-1.3, mean dose 1.9 mg). As shown in the Neuropsychiatric Inventory, both medications reduced delusion and agitation, whereas quetiapine additionally improved depression and anxiety. Haloperidol worsened aberrant motor behaviour and caused extrapyramidal symptoms. In the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) neuropsychological test battery which assessed cognitive parameters, quetiapine improved word recall; significant interaction terms revealed differences between quetiapine and haloperidol in word-list memory and constructional praxis. According to the Nurses' Observation Scale for Geriatric Patients (NOSGER) quetiapine improved instrumental activities of daily living. Actimetry documented the circadian rest-activity cycle before and after treatment. Sleep analysis revealed that patients receiving quetiapine had shorter wake bouts during the night, whereas patients receiving haloperidol had fewer though longer immobile phases. The study provides evidence that quetiapine at a moderate dose may be efficacious in treating behavioural disturbances in AD, with better tolerability than haloperidol. PMID- 16316486 TI - Divalproex sodium vs. placebo in the treatment of repetitive behaviours in autism spectrum disorder. AB - Autism is a neurodevelopmental disorder characterized by impairment in three core symptom domains: socialization, communication, and repetitive/stereotyped behaviours. Other associated symptom domains are also affected including impulsivity/aggression, self-injury, anxiety, and mood lability. Divalproex has been shown to have efficacy in treating epilepsy, bipolar disorder, mood lability, and impulsive aggression. The present study evaluated the use of divalproex in the treatment of repetitive, compulsive-like symptoms of autism spectrum disorder (ASD). Thirteen individuals with ASD participated in an 8-wk, double-blind, placebo-controlled trial of divalproex sodium vs. placebo. There was a significant group difference on improvement in repetitive behaviours as measured by the Children's Yale-Brown Obsessive Compulsive Scale (C-YBOCS) (p=0.037) and a large effect size (d=1.616). This study provides preliminary support for the use of divalproex in treating repetitive behaviours in ASD. Further research is needed to evaluate the specificity and mechanism of action of these findings. PMID- 16316487 TI - Clozapine and olanzapine, but not haloperidol, suppress serotonin efflux in the medial prefrontal cortex elicited by phencyclidine and ketamine. AB - N-methyl-D-aspartate (NMDA) receptor antagonists such as phencyclidine (PCP) and ketamine can evoke psychotic symptoms in normal individuals and schizophrenic patients. Here, we have examined the effects of PCP (5 mg/kg) and ketamine (25 mg/kg) on the efflux of serotonin (5-HT) in the medial prefrontal cortex (mPFC) and their possible blockade by the antipsychotics, clozapine, olanzapine and haloperidol, as well as ritanserin (5-HT2A/2C receptor antagonist) and prazosin (alpha1-adrenoceptor antagonist). The systemic administration, but not the local perfusion, of the two NMDA receptor antagonists markedly increased the efflux of 5-HT in the mPFC. The atypical antipsychotics clozapine (1 mg/kg) and olanzapine (1 mg/kg), and prazosin (0.3 mg/kg), but not the classical antipsychotic haloperidol (1 mg/kg), reversed the PCP- and ketamine-induced increase in 5-HT efflux. Ritanserin (5 mg/kg) was able to reverse only the effect of PCP. These findings indicate that an increased serotonergic transmission in the mPFC is a functional consequence of NMDA receptor hypofunction and this effect is blocked by atypical antipsychotic drugs. PMID- 16316488 TI - Molecular and morphological characterization of Echinococcus in cervids from North America. AB - Many issues concerning the taxonomy of Echinococcus have been resolved in recent years with the application of molecular tools. However, the status of Echinococcus maintained in transmission cycles involving cervid intermediate hosts remains to be determined. The recent characterization of the parasite from cervids in Finland has highlighted the paucity of data available, particularly that from North America. In this study, we have characterized a large number of Echinococcus isolates from cervids from Western Canada on the basis of morphology and molecular genetic techniques. Our results support earlier studies suggesting that Echinococcus of cervid origin is phenotypically and genetically distinct to Echinococcus maintained in domestic host assemblages, and also confirms that Echinococcus of cervid origin does not constitute a genetically homogeneous group. However, our data do not support the existence of 2 distinct genotypes (strains/subspecies) with separate geographical distributions. Our data appear to support the existence of only 1 species in cervids, but additional isolates from cervids and wolves in other endemic regions should be characterized before a final decision is made on the taxonomic status of Echinococcus in cervids. PMID- 16316489 TI - A modelling analysis of pertussis transmission and vaccination in Rio de Janeiro, Brazil. AB - Pertussis is an infectious respiratory disease for which mass vaccination is an effective preventive strategy. In many developed countries, where high vaccination coverage has been maintained for approximately 50 years, re-emergence of the disease has been observed in all age groups. In the municipality of Rio de Janeiro (RJ), where vaccination started in the 1980s, surveillance data show no sign of disease re-emergence. We developed a mathematical model that incorporates the major demographic aspects of a large urban centre in a developing nation, in addition to the most important epidemiological aspects of disease transmission. Parameter values were estimated based on RJ demographic and vaccine coverage data. Overall, all vaccination strategies determined a major decrease (over 95% decrease when compared to the pre-vaccine era) in the incidence of primary infections (occurring in individuals who have never been immunized through infection or vaccine). On the other hand, the strategies (a) three doses at age 2 11 months, (b) three doses plus booster at age 12-23 months, (c) three doses plus booster at age 4-5 years, and (d) three doses plus both boosters, differently affected the incidence of secondary infections (occurring in previously infected/vaccinated individuals). Given that the immunity against pertussis wanes with time and that the infectious agent has not been eliminated from the population, it is expected that pertussis will continue to be a problem in RJ. Actually, since immunity acquired from vaccine wanes faster than disease-acquired immunity and the possibility of natural boosters has decreased with mass vaccination, an increase in the incidence of secondary infections among older age groups is expected (and predicted by the model). Possible explanations as to why this dynamics is not captured by the RJ surveillance system are discussed. A poorly effective surveillance system and a lack of awareness regarding loss of immunity and the possibility of pertussis infection in older age groups are among them. Finally, we bring attention to the need of (i) field studies for the measurement of pertussis incidence in adolescents and adults; (ii) better understanding of the transmission dynamics currently occurring in RJ, and (iii) re-evaluation of vaccination strategies with the possible introduction of acellular vaccines for the vaccination of older individuals. PMID- 16316490 TI - Most Campylobacter subtypes from sporadic infections can be found in retail poultry products and food animals. AB - The subtypes of Campylobacter isolates from human infections in two Danish counties were compared to isolates from retail food samples and faecal samples from chickens, pigs and cattle. During a 1-year period, 1285 Campylobacter isolates from these sources were typed by two methods: 'Penner' heat-stable serotyping and automated ribotyping (RiboPrinting). C. jejuni was the dominating species, but C. coli was more prevalent among food and chicken isolates (16%) compared to human isolates (4%). In total, 356 different combined sero-ribotypes (subtypes) were found. A large subtype overlap was seen between human isolates and isolates from food (66%), chickens (59%) and cattle (83%). This was verified by PFGE typing of 212 isolates representing selected subtypes. All frequent (n>3) subtypes found in food were also present in humans. Sixty-one per cent of the isolates from domestically acquired infections had subtypes that were also found in food as opposed to 31% of travel-associated infections. The results showed differences in the various Campylobacter populations, e.g. the Danish population as reflected in the domestically acquired infections and the Danish-produced food was more uniform than the isolates originating from outside the country. The study shows that most C. jejuni subtypes found in poultry food samples, broiler chickens, and cattle were represented in the domestically acquired cases, indicating that C. jejuni from these reservoirs are likely sources of human infections in Denmark. PMID- 16316491 TI - Exposure to school children as a risk factor in a community outbreak of hepatitis A in young adults: a case control study. AB - To investigate risk factors during a community outbreak of hepatitis A we carried out a case- control study of 35 cases and 49 matched controls using an interviewer-administered questionnaire on clinical history, travel, household details including domestic toilet facilities, infectious contacts, and food history. Of 99 cases notified in the city during the outbreak year, 50 (51%) were young adults age 15-34 years. Hepatitis A infection was independently associated with household contact with a case (P=0.0005), and sharing a household with children in primary school (OR 3.4, 95% CI 1.2-9.5, P=0.008) with risk increasing with number of primary-school pupils in the household (chi(2) for linear trend 6.47, P=0.01). We concluded that in a population with a low prevalence of hepatitis A, adults who live in the same household as primary-school-age children are at increased risk of acquiring the infection during community outbreaks. PMID- 16316492 TI - Seroepidemiology of diphtheria and pertussis in Luxembourg in 2000. AB - A large serosurvey was carried out in Luxembourg in 2000-2001, to determine the population immunity against a number of vaccine-preventable infections including diphtheria and pertussis. Immunity to diphtheria and pertussis was assessed using an in-house neutralization assay and a commercial ELISA test respectively. Mean pertussis antibody activity decreased from 4 to 8 years of age, reflecting the effects of waning of vaccine-induced immunity. Mean pertussis antibody activity increased during adolescence due to infection in previously vaccinated individuals and levelled out after approximately 20 years of age. For adults>25 years age, a statistically significant 30% difference in mean antibody activity between men and women was observed. The proportion of seronegatives for diphtheria among children and adolescents aged<20 years was 2.5% reflecting the high vaccination coverage. The proportion seronegative for diphtheria tended to increase with age such that 42% of individuals aged>40 years were seronegative. Our study supports the recently introduced acellular pertussis vaccine booster at 6 years to reduce pertussis transmission in school-aged children and adolescents. PMID- 16316493 TI - Detailed analysis of the genetic evolution of influenza virus during the course of an epidemic. AB - The genetic variability of influenza virus is usually studied with sequences selected over numerous years and countries, and rarely within a single season. Here we examined the viral evolution and the correlation between genetic and clinical features during an epidemic. From a French prospective household-based study in 1999-2000, 99 infected patients were randomly selected. The HA1 genomic domain was sequenced. Phylogenetic analysis showed the existence of two groups of A/H3N2 viruses. We found no distinct pattern of genomic evolution within either group according to time. A spatial correlation with the nucleotide distances was shown. The average nucleotide diversity was 3.4x10-3 nucleotides per site, and did not differ between the groups. A lower number of segregating sites was observed in patients who experienced influenza-like symptoms during the previous epidemic. These results suggest that the influenza virus undergoes regular HA1 nucleotide changes, but without clonal expansion of mutant strains within a single epidemic. PMID- 16316494 TI - The effect of mass influenza immunization in children on the morbidity of the unvaccinated elderly. AB - The objectives of these studies were to analyse the effect of mass influenza immunization in children on the morbidity of unvaccinated non-institutionalized elderly during an influenza epidemic. A mass vaccination campaign with vaccine was conducted in children aged 3-6 years attending kindergartens (57.4% of 6374) and aged 7-17 years attending schools (72% of 34237) in two communities of the Moscow region. The clinical effectiveness of vaccination was 60.9% for kindergartens and 68.8% for schools. There were 3.4 times fewer episodes of influenza-like illnesses and 1.7-2.6 fewer episodes in all seven diseases which are possible complications of influenza out of the 10 evaluated diseases in 158451 unvaccinated non-institutionalized elderly people during the influenza epidemic compared with the control communities. The differences were found to be statistically significant. Mass vaccination of children attending child institutions brought about a significant reduction of both influenza-like illnesses in children and influenza-associated illnesses in unvaccinated non institutionalized elderly persons living in the home setting. PMID- 16316495 TI - Chlamydia pneumoniae and Mycoplasma pneumoniae pneumonia: comparison of clinical, epidemiological characteristics and laboratory profiles. AB - The purpose of our retrospective 3-year study was to analyse and compare clinical and epidemiological characteristics in hospitalized patients older than 6 years with community-acquired pneumonia (CAP) caused by Chlamydia pneumoniae (87 patients) and Mycoplasma pneumoniae (147 patients). C. pneumoniae and M. pneumoniae infection was confirmed by serology. C. pneumoniae patients were older (42.12 vs. 24.64 years), and were less likely to have a cough, rhinitis, and hoarseness (P<0.001). C. pneumoniae patients had higher levels of C-reactive protein (CRP), and aspartate aminotransferase (AST) than M. pneumoniae patients (P<0.001). Pleural effusion was recorded more frequently in patients with M. pneumoniae (8.84 vs. 3.37%). There were no characteristic epidemiological and clinical findings that would distinguish CAP caused by M. pneumoniae from C. pneumoniae. However, some factors are indicative for C. pneumoniae such as older age, lack of cough, rhinitis, hoarseness, and higher value of CRP, and AST. PMID- 16316496 TI - Serological evidence of West Nile Virus (WNV) in mammalian species in Turkey. AB - In this study, the sera collected from a variety of mammalian species (ass-mules, cat, cattle, dog, horse, human and sheep) in 10 representative provinces of Turkey, were surveyed for the presence of neutralizing antibodies to West Nile virus (WNV). Overall, 1 of 40 (2.5%) ass-mules, 4 of 100 (4%) cattle, 43 of 114 (37.7%) dogs, 35 of 259 (13.5%) horses, 18 of 88 (20.4%) humans and 1 of 100 (1%) sheep, tested positive for WNV-neutralizing antibodies. The results indicate that a wide range of mammals are exposed to a West Nile-related virus and this could contribute to the long-term survival of this virus in the absence of overt disease. PMID- 16316497 TI - Epidemiological findings and medical, legal, and public health challenges of an investigation of severe soft tissue infections and deaths among injecting drug users -- Ireland, 2000. AB - In May 2000, public health authorities in Dublin, Ireland, identified a cluster of unexplained severe illness among injecting drug users (IDUs). Similar clusters were also reported in Scotland and England. Concurrent investigations were undertaken to identify the aetiology and source of the illnesses. In Dublin, 22 IDUs were identified with injection-site inflammation resulting in hospitalization or death; eight (36%) died. Common clinical findings among patients with severe systemic symptoms included leukaemoid reaction and cardiogenic shock. Seventeen (77%) patients reported injecting heroin intramuscularly in the 2 weeks before illness. Of 11 patients with adequate specimens available for testing, two (18%) were positive by 16S rDNA PCR for Clostridium novyi. Clinical and laboratory findings suggested that histotoxic Clostridia caused a subset of infections in these related clusters. Empiric treatment for infections among IDUs was optimized for anaerobic organisms, and outreach led to increased enrolment in methadone treatment in Dublin. Many unique legal, medical, and public health challenges were encountered during the investigation of this outbreak. PMID- 16316505 TI - Analysis of organic films and interfacial layers by infrared spectroscopic ellipsometry. PMID- 16316506 TI - Off-line direct deposition gas chromatography/surface-enhanced Raman scattering and the ramifications for on-line measurements. AB - Surface-enhanced Raman spectra (SERS) of molecules separated by gas chromatography (GC) were measured off-line by condensing the analyte on a moving, liquid-nitrogen-cooled ZnSe window on which a 5 nm layer of silver had been formed by physical vapor deposition. After the components that eluted from the chromatograph had been deposited, the substrate was allowed to warm up to room temperature and transferred to the focus of a Raman microspectrometer where the spectrum of each component was measured. Band intensities in the spectrum of 3 ng of caffeine prepared in this way were approximately the same as in the spectrum of bulk caffeine. By making some logical assumptions, it was shown that identifiable GC/SERS spectra of 30 pg of many molecules could be measured over a 300 cm(-1) region in real-time and that if an optimized substrate were used the minimum identifiable quantity could be reduced to 1 pg or less. PMID- 16316507 TI - High-speed gas chromatography-multiplex coherent Raman analysis of BTEX. AB - Gas chromatography-multiplex coherent Raman (GC-MCR) is a new tandem technique that can be used for the high-speed analysis of volatile mixtures. BTEX serves as a useful and challenging test sample because of the similarity in boiling point and spectroscopic properties of its constituents. The ability to spectroscopically resolve isomers (e.g., m-xylene and p-xylene) allows GC-MCR to sacrifice chromatographic resolution for speed. The result is the analysis of BTEX in less than 5 min, which is relatively fast compared with other tandem GC techniques. PMID- 16316508 TI - Read-noise characterization of focal plane array detectors via mean-variance analysis. AB - Mean-variance analysis is described as a method for characterization of the read noise and gain of focal plane array (FPA) detectors, including charge-coupled devices (CCDs), charge-injection devices (CIDs), and complementary metal-oxide semiconductor (CMOS) multiplexers (infrared arrays). Practical FPA detector characterization is outlined. The nondestructive readout capability available in some CIDs and FPA devices is discussed as a means for signal-to-noise ratio improvement. Derivations of the equations are fully presented to unify understanding of this method by the spectroscopic community. PMID- 16316509 TI - Raman and near-infrared spectroscopy for quantification of fat composition in a complex food model system. AB - Raman and near-infrared (NIR) spectroscopy have been evaluated for determining fatty acid composition and contents of main constituents in a complex food model system. A model system consisting of 70 different mixtures of protein, water, and oil blends was developed in order to create a rough chemical imitation of typical fish and meat samples, showing variation both in fatty acid composition and in contents of main constituents. The model samples as well as the pure oil mixtures were measured using Raman and NIR techniques. Partial least squares regression was utilized for prediction, and fatty acid features were expressed in terms of the iodine value and as contents of saturated, monounsaturated, and polyunsaturated fatty acids. Raman spectroscopy provided the best results for predicting iodine values of the model samples, giving validated estimation errors accounting for 2.8% of the total iodine value range. Both techniques provided good results for predicting the content of saturated, monounsaturated, and polyunsaturated fatty acids in the model samples, yielding validated estimation errors in the range of 2.4-6.1% of the total range of fatty acid content. Prediction results for determining fatty acid features of the pure oil mixtures were similar for the two techniques. NIR was clearly the best technique for modeling content of main constituents in the model samples. PMID- 16316510 TI - Forensic analysis of architectural finishes using fourier transform infrared and Raman spectroscopy, part I: the resin bases. AB - The ability of Raman spectroscopy and Fourier transform infrared (FT-IR) microscopy to discriminate between resins used for the manufacture of architectural finishes was examined in a study of 39 samples taken from a commercial resin library. Both Raman and FT-IR were able to discriminate between different types of resin and both split the samples into several groups (six for FT-IR, six for Raman), each of which gave similar, but not identical, spectra. In addition, three resins gave unique Raman spectra (four in FT-IR). However, approximately half the library comprised samples that were sufficiently similar that they fell into a single large group, whether classified using FT-IR or Raman, although the remaining samples fell into much smaller groups. Further sub division of the FT-IR groups was not possible because the experimental uncertainty was of similar magnitude to the within-group variation. In contrast, Raman spectroscopy was able to further discriminate between resins that fell within the same groups because the differences in the relative band intensities of the resins, although small, were larger than the experimental uncertainty. PMID- 16316511 TI - Forensic analysis of architectural finishes using fourier transform infrared and Raman spectroscopy, part II: white paint. AB - White household paints are commonly encountered as evidence in the forensic laboratory but they often cannot be readily distinguished by color alone so Fourier transform infrared (FT-IR) microscopy is used since it can sometimes discriminate between paints prepared with different organic resins. Here we report the first comparative study of FT-IR and Raman spectroscopy for forensic analysis of white paint. Both techniques allowed the 51 white paint samples in the study to be classified by inspection as either belonging to distinct groups or as unique samples. FT-IR gave five groups and four unique samples; Raman gave seven groups and six unique samples. The basis for this discrimination was the type of resin and/or inorganic pigments/extenders present. Although this allowed approximately half of the white paints to be distinguished by inspection, the other half were all based on a similar resin and did not contain the distinctive modifiers/pigments and extenders that allowed the other samples to be identified. The experimental uncertainty in the relative band intensities measured using FT IR was similar to the variation within this large group, so no further discrimination was possible. However, the variation in the Raman spectra was larger than the uncertainty, which allowed the large group to be divided into three subgroups and four distinct spectra, based on relative band intensities. The combination of increased discrimination and higher sample throughput means that the Raman method is superior to FT-IR for samples of this type. PMID- 16316512 TI - Hydrogen-deuterium exchange in bovine serum albumin protein monitored by fourier transform infrared spectroscopy, part I: structural studies. AB - The structure of the amide I band of bovine serum albumin (BSA) was determined using an H/D exchange experiment. The difference between the dry and hydrated exchange spectrum revealed the fine structure of the amide I band. The band at 1717 +/- 2 cm(-1) is due to the vibration of the COOH moieties from the protein side chains. Band components at 1682 +/- 2 cm(-1), 1655 +/- 2 cm(-1), and 1637 +/ 2 cm(-1) are assigned to the vibrations of the backbone C=O. These three bands belong to vibrations of three different populations of amide groups differing in the number of established H-bonds. The connectivity between the frequencies of various amide vibrations was determined by two-dimensional generalized correlation spectroscopy and spectral decomposition. About 7% of the whole exchangeable hydrogen atom population (NH, NH2, and OH groups from backbone and side chains) remains unexchanged, and these hydrogen atoms belong mainly to the NH groups, which are H-bonded to specific C=O groups. Moreover, this study concerns the approximately 10% of hydrogen atoms belonging to a particular HN...O=C population with a characteristic amide A frequency at 3290 cm(-1) and an amide I band at 1655 +/- 2 cm(-1), usually attributed to the alpha-helical structure that remains unexchanged. At higher temperature the exchange is more efficient. Upon heating, a further 4% of these NH groups are deuterated. The comparison of the exchange spectrum at higher temperature with the structural changes of the protein at the same temperature implies that the change in overall dynamics of the protein improve the level of exchange. PMID- 16316513 TI - Hydrogen-deuterium exchange in bovine serum albumin protein monitored by Fourier transform infrared spectroscopy, part II: kinetic studies. AB - The set of infrared spectra recorded at different levels of hydrogen-deuterium (H/D) exchange as a function of time were processed using spectral decomposition. The most precise information about H/D exchange of the NH groups of the protein backbone was retrieved by observing the intensity change of the nearest C=O stretching vibration. The H/D exchange at the protein backbone begins with NH groups bonded to C=O with a characteristic frequency of 1683 cm(-1). These amide groups were initially free and are the first to accept H-bonds from water during the hydration process. The NH groups, which are connected to C=O groups with a characteristic band at 1657 cm(-1), exchange at a slower rate. For both populations the pairs of comparable exchange rates were calculated with rate constants of 10(-3) min(-1), 0.014 min(-1), 0.0046 min(-1), and 0.09 min(-1). The appearance of two different exchange rates for each population is a consequence of the distinct exposure of particular molecular groups to the solvent. Two additional bands sensitive to exchange are attributed to NH bending modes in the side chains and are located at 1610 cm(-1) and 1585 cm(-1). These NH groups undergo H/D exchange at the beginning of the exchange with exchange rates of 0.019 min(-1) and 0.17 min(-1), respectively. The calculated exchange rate for hydrating water molecules is 0.037 min(-1). The distribution of water's exchange rate is extremely broad and covers almost the entire interval of the time dependent experiment. From the efficiency of the exchange (96%) it is evident that some parts of the protein are completely forbidden to water molecules. PMID- 16316514 TI - Determination of the polymorphic forms of bicifadine hydrochloride by differential scanning calorimetry-thermogravimetric analysis, X-ray powder diffraction, attenuated total reflectance-infrared spectroscopy, and attenuated total reflectance-near-infrared spectroscopy. AB - The pharmaceutical compound bicifadine hydrochloride, which has been found to crystallize in two polymorphic forms, has been characterized by thermal analysis, X-ray powder diffraction (XRPD), infrared (IR) spectroscopy, and near-infrared (NIR) spectroscopy. A series of 22 sample mixtures of polymorph 1 and polymorph 2 were prepared and calibration models for the quantitation of these binary mixtures have been developed for each of the XRPD, attenuated total reflectance (ATR)-IR, and ATR-NIR analytical techniques. The quantitative results were obtained using a partial least squares (PLS) algorithm, which predicted the concentration of polymorph 1 from the XRPD spectra with a root mean standard error of prediction (RMSEP) of 4.4%, from the IR spectra with a RMSEP of 3.8%, and from the NIR spectra with a RMSEP of 1.4%. The studies indicate that when analyses are carried out on equivalent sets of spectra, NIR spectroscopy offers significant advantages in quantitative accuracy as a tool for the determination of polymorphs in the solid state and is also more convenient to use than both the ATR-IR and XRPD methods. Density functional theory (DFT) B3LYP calculations and IR spectral simulation have been used to determine the nature of the vibrational modes that are the most sensitive in the analysis. PMID- 16316515 TI - Multicomponent peak modeling of protein secondary structures: comparison of gaussian with lorentzian analytical methods for plant feed and seed molecular biology and chemistry research. AB - The objective of this study was to compare Gaussian and Lorentzian multicomponent peak modeling methods in quantification of protein secondary structures of various plant seed and feed tissues within intact tissue at a cellular and subcellular level using the advanced synchrotron light sourced Fourier transform infrared (FT-IR) microspectroscopy (S-FTIR). This experiment was performed at the beamline U10B at the National Synchrotron Light Source (NSLS) in Brookhaven National Laboratory (BNL), U.S. Dept of Energy (NSLS-BNL, NY). The results show that in the comparison of the Gaussian and Lorentzian multi-peak modeling methods, the Gaussian method is more accurate for fitting multi-peak curves of protein secondary structures than the Lorentzian method, with higher modeling R(2) values (0.92 versus 0.89, P < 0.05). There were no large differences (P > 0.05) in the quantification of the relative percentage alpha-helices, beta sheets, and others in protein secondary structures of the plant seed tissues, with averages of 30.2%, 40.4%, and 29.4%, respectively. However, there are significant differences (P < 0.05) in the quantification of the ratios of sheet alpha-helix (1.42 versus 1.60; SEM = 0.058) in protein secondary structures of the plant seed tissues. With synchrotron FT-IR microspectroscopy, the ultrastructural-chemical makeup and nutritive characteristics could be revealed at a high spatial resolution. Synchrotron-based FT-IR microspectroscopy revealed that the secondary structure of protein differed between the plant seed tissues in terms of relative percentage and ratio of protein secondary structures (alpha helix and beta-sheet) within cellular dimensions. The results also show that the flaxseed tissues contained higher (P < 0.05) percentage alpha-helix (38.6 versus 24.0%) beta-sheet (45.3 versus 36.9%), lower (P < 0.05) percentage of other secondary structures (16.1% versus 39.0%), and higher (P < 0.05) ratios alpha helix beta-sheet (0.90 versus 0.69) than the winterfat seed tissues. It must be mentioned that the relative percentages of protein secondary structure may not reflect the true secondary structure. However, the purpose of modeling the relative percentage of secondary structure was to detect the variety of differences among seed/feed/plant tissues and their relation to nutritive value and digestive behavior. The results demonstrate the potential of highly spatially resolved synchrotron-based FT-IR microspectroscopy to reveal protein secondary structures of the plant seed/feed tissues. Further study is needed to quantify the relationship between protein secondary structures and nutrient availability and digestive behavior of various varieties of plant seed tissues. Information from the infrared probing of protein secondary structures can be valuable as a guide to maintaining protein nutritive value and quality for animal and human use. PMID- 16316516 TI - Scatter correction of transmission near-infrared spectra by photon migration data: quantitative analysis of solids. AB - The scope of this work is a new methodology to correct conventional near-infrared (NIR) data for scattering effects. The technique aims at measuring the absorption coefficient of the samples rather than the total attenuation measured in conventional NIR spectroscopy. The main advantage of this is that the absorption coefficient is independent of the path length of the light inside the sample and therefore independent of the scattering effects. The method is based on time resolved spectroscopy and modeling of light transport by diffusion theory. This provides an independent measure of the scattering properties of the samples and therefore of the path length of light. This yields a clear advantage over other preprocessing techniques, where scattering effects are estimated and corrected for by using the shape of the measured spectrum only. Partial least squares (PLS) calibration models show that, by using the proposed evaluation scheme, the predictive ability is improved by 50% as compared to a model based on conventional NIR data alone. The method also makes it possible to predict the concentration of active substance in samples with other physical properties than the samples included in the calibration model. PMID- 16316517 TI - Analysis of sticky cotton by near-infrared spectroscopy. AB - "Stickiness" in cotton is a major problem affecting throughput in cotton gins and spinning mills alike. Stickiness is thought to be caused by the deposition of sugars by insects, principally aphid and whitefly, on the open boll. Fourier transform near-infrared (FT-NIR) spectroscopy was used to develop models for sugar content from high-pressure liquid chromatography (HPLC), thermodetector, and mini-card data. A total of 457 cotton samples were selected to represent both Upland and Pima varieties and cotton processing before and after ginning. The Unscrambler was used to develop the models. A successful model was made to determine the mini-card value and successfully detect "stickiness". The standard error of cross-validation (SECv) was 0.26 with an R2 of 0.96. The model was not improved by increasing the range of "stickiness" as measured by the mini-card from the usual 0-3 scale to a scale of 0-8. If a value is determined to be greater than 1 it will be difficult to blend bales at a spinning plant "opening line" to allow for maximum efficiency of spinning. PMID- 16316518 TI - Near-infrared combination and overtone bands of CH in CHX3, CHX(2)-CHX2, and CHX(2)-CX(2)-CHX2. AB - In the present report we studied spectral characteristics of the near-infrared combination and overtone bands of CH vibrations of a CH sequence. The near infrared bands of the CH in CHX3 (X, halogen), which were interpreted in terms of the CH stretching and CH deformation fundamentals without any ambiguity, typically showed how the frequency and intensity of a combination or an overtone depend on the vibrational excited state. In the CH-C-CH of CHX2CX2CHX2, the vibrations of one CH are isolated from those of the other CH, and the combination and overtone bands were similarly interpreted as those of the CH, although each of the combination bands was split into two because of non-degeneracy of the CH deformation. In the CH-CH of CHX2CHX2, the CH deformations only have coupled modes. The first combination showed four narrowly separate bands, which were reasonably interpreted on the basis of the CH stretching and the coupled CH deformation modes. We demonstrated that the first combination of coupled modes as well as the combination of up to, at least, the third order of isolated modes have the nature of the characteristic bands. PMID- 16316519 TI - Visible and near-infrared calibrations for quality assessment of fresh phase I and II mushroom (Agaricus bisporus) compost. AB - Previous studies have shown that visible and near-infrared spectra (Vis-NIR) of dry and milled compost can be used for generating partial least squares (PLS) calibrations of phase II compost parameters including ammonia, nitrogen dry matter (NDM), dry matter (DM), pH, conductivity, carbon, microbial population, and potential productivity. The objective of this study was to develop robust calibrations for some of the key parameters from the spectra of fresh phase I and II composts. Samples of substrates from six commercial production yards were obtained during winter and summer months of 2000-2004 to monitor changes in quality and were analyzed for the test factors. Vis-NIR reflectance measurements of fresh samples (740) were made over the range of 400-2500 nm. After mathematical pretreatments, PLS calibrations of the key parameters were developed using the NIR (1100-2500 nm) and visible and NIR (400-2500 nm) regions and subsequently validated using an independent sample set of 123 phase I and II samples obtained during 2004-2005. The phase I and II standard errors of laboratory measurements of ammonia, pH, conductivity, DM, NDM, and ash were lower than the standard error of predictions of the same parameters, respectively, by the best NIR or Vis-NIR models. The degree of precision for some of the calibrations, especially ammonia, NDM, and DM, is suitable for composters to monitor changes in quality parameters during production. The laboratory measurement errors for phase I samples were greater than those of the phase II samples, except for ash, due to a higher degree of heterogeneity in the substrate. The calibrations, especially for pH, conductivity, and ash, need to be improved with new sample sets. A major advantage of NIR spectroscopy is the ability to assess substrate quality for a range of target parameters simultaneously, within a few hours of receiving the samples. The main drawbacks are the expensive instrumentation, expertise, and training necessary for operating the spectrometer and a dedicated chemometrician required for maintaining the equations compared to the reference methods. PMID- 16316520 TI - Partial least squares based decomposition of five spectrally overlapping factors. AB - The development of a multi-sensory fiber-optic based fluence rate probe (MSP) for light monitoring and dosimetry during photodynamic therapy (PDT) created the need for a robust multivariate signal analysis algorithm capable of quantifying the intensity of five component spectra, representing the sensors, which display a large degree of spectral overlap. Partial least squares (PLS) analysis, as an option for such an analysis algorithm, was evaluated through simulations in the presence of three types of noise, which experimentally may limit the accuracy of PLS quantification of component spectra contributions. Random, or white noise, background was varied over a range of 0-15% relative intensity. A non-random (Gaussian) background vector was simulated, as an experimentally relevant spectral interferent, over a range of 0-7% relative intensity. Spectral variation was modeled by a spectral shift of +/-1-5 nm. Procedures for selecting preferred combinations of fluorophores, with minimum possible spectral overlap, were developed. To illustrate the importance of this selection process, PLS performance results were compared for two possible combinations of five fluorophores, as well as for the combination of three fluorophores currently in experimental use with MSPs. Experimentally anticipated worst-case quantifications were identified for all three types of simulated noise (5% random background, 0.5% Gaussian background, and +/-2 nm spectral shift). The effects of single and combined types of noise were evaluated by independently varying each type of simulated noise over the experimentally relevant range. The mean percentage error in determining the fluorophore contributions and hence quantification of the fluence rate were compared with the required performance standard of better than 10% error for optical power meters in medical use. The PLS algorithm provided an accuracy of 7 +/- 2% for five fluorophores and 3.3 +/- 0.8% for three fluorophores, indicating that PLS is an appropriate choice for this application. PMID- 16316521 TI - In situ high-temperature visible microspectroscopy for volcanic materials. AB - In situ high-temperature visible microspectroscopy has been developed in order to study color change kinetics of volcanic materials. Olivine thin sections put on a synthetic alumina plate are heated on a heating stage at 600-800 degrees C under a visible microspectroscope. Changes in visible absorption spectra are monitored every 60 s for 5 hours. The obtained high-temperature visible spectra showed a gradual increase with time in absorbance in the shorter wavelength region (400 600 nm). The 430 nm absorbance (ligand field transition of Fe3+ increased more with time at higher temperatures. Assuming diffusional transport in plane sheets, apparent diffusion coefficients were determined at temperatures of 600-800 degrees C. The activation energy for this diffusion in olivine is 208 +/- 17 kJ/mol. This activation energy value is similar to those for the metal vacancy diffusion in olivine. This newly developed in situ high-temperature visible microspectroscopy can provide kinetic measurements of visible spectral change of materials at high temperatures such as volcanic materials. PMID- 16316522 TI - Observation of large photoacoustic signal phase changes during a diffusion process. AB - The phase of the photoacoustic signal is known to be a sensitive and accurate means to investigate, both qualitatively and quantitatively, static multilayer heterogeneous systems. According to theory, the maximum phase delay for a very weakly absorbing homogeneous sample should be within 45 degrees of a very strongly absorbing sample, while for heterogeneous samples the phase delay can be greater than 45 degrees. Here we report the observation of photoacoustic phase delays greater than 350 degrees by extending the use of step-scan phase modulation photoacoustic spectroscopy to study a non-repetitive dynamic system in situ, in real time. These large phase delays correspond to sampling several thermal diffusion lengths into the sample. The model system used in this study consisted of a hydrocarbon grease diffusing through a porous Teflon film. The progress of the diffusion was tracked by monitoring both the photoacoustic signal magnitude and the phase of the hydrocarbon grease after isolation from the Teflon film signal contributions at two different phase modulation frequencies. PMID- 16316523 TI - [Certain problems in diagnosis and treatment of chronic renal diseases]. PMID- 16316524 TI - [NPHS1 mutations in a Chinese family with congenital nephrotic syndrome]. AB - OBJECTIVE: Congenital nephrotic syndrome (CNS) is defined as heavy proteinuria or nephrotic syndrome occurring before 3 months of age. It is characterized by early onset, resistance to steroid therapy and progressing to end-stage renal disease (ESRD). In recent years, several genes associated with CNS have been identified, such as NPHS1, NPHS2 and WT1. The mutations of these genes have been identified in the patients with CNS in Finland, other European countries, North Africa, North America, and Asia, respectively. However, the investigation of the above genes has not been performed in Chinese CNS patients. In this study, NPHS1 mutations in a Chinese family with CNS were detected and analyzed. METHODS: There were two CNS patients in the investigated family. The proband, a 45-day-old boy, was born at fullterm and weighed 2700 g at birth. The placenta weighed 450 g. At the age of 10 days, generalized edema, proteinuria, hypoproteinemia, and hypoalbuminemia were found without renal insufficiency. The proband's sister, with the same phenotype and normal renal function, underwent renal biopsy at 5 years of age. Their parents and elder half-sister all had normal phenotypes. Genomic DNA samples were extracted from peripheral bloods of the proband, his family members and 50 unrelated, normal individuals. All 29 exons and exon-intron boundaries of NPHS1 were detected in the proband by polymerase chain reaction (PCR), direct DNA sequencing, and restriction enzyme analysis. RESULTS: Three heterozygous mutations of NPHS1, namely, G928A (D310N), 1893-1900del 8 (CGAAACCG), and G2869C (V957L) were identified in the proband. These mutations involved exons 8, 14, and 21. The same genotype was found in the proband's sister who had the same phenotype, but was not detected in proband's elder half-sister who had normal phenotype. Fifty normal individuals had no these mutations. The proband's mother with normal urinalysis had G928A (D310N) heterozygous mutation, and the father with normal urinalysis had two heterozygous mutations of 1893 1900del 8 (CGAAACCG) and G2869C (V957L). At the same time, three types of single nucleotide polymorphisms (SNPs), E117K (rs3814995), S1105S (rs2071327), and IVS27+45c > t, were confirmed in the proband. Another variant, IVS8+68 a > g had also been found. CONCLUSION: This is the first report about NPHS1 mutations in Chinese CNS kindred. These three heterozygous mutations of NPHS1 are novel genetic defects of CNS, which have not been described before. PMID- 16316525 TI - [Renal impairment in patients with methylmalonic aciduria: a review of five cases]. AB - OBJECTIVE: The renal impairment in children with methylmalonic aciduria has seldom been reported. To improve knowledge in this aspect, clinical data of five cases with methylmalonic aciduria with renal involvement were analyzed and the results are reported in this paper, which may be of some help in early diagnosis, treatment and in achieving favorable prognosis. METHODS: Urine methylmalonic acid was measured by gas chromatography-mass spectrometry analysis, if the content exceeded the normal range and vitamin B12 deficiency was excluded, the diagnosis of methylmalonic aciduria was confirmed. Homocysteine in plasma was also measured with fluorescence polarization immunoassay to make sure if concomitant homocysteinemia existed. From January 2002 to January 2005, five patients who had renal impairment were diagnosed as methylmalonic aciduria by urinary organic acid analysis. Among them, three were male, two were female, aged from seven months to 26 years, with average of 13 years. Three were presented to pediatric nephrology clinic with hematuria, proteinuria or edema, the other two were presented to pediatric neurology clinic first for psychomotor retardation. Their clinical features, laboratory findings, treatment regimens and prognosis were analyzed and summarized. RESULTS: All the five patients with methylmalonic aciduria were found to have various degrees of renal impairment, manifested as hematuria or proteinuria. Among them, two cases had gross hematuria and three had microscopic hematuria. Edema was found in two cases and hypertension occurred in one case. Early indicators of renal damage, such as microalbunminuria, N-acetyl-beta-D glucosaminidase, transferrin and alpha-microglobulin showed glomerular and tubular dysfunction. Clinically nephrotic syndrome was diagnosed in one case, the other four cases were diagnosed as glomerulonephritis, and two cases had renal failure. Renal biopsy was performed in one case, tubulo-interstitial damage and mesangial proliferation appeared. Mental retardation and psychomotor disorder were chief nervous system complaints. Leukodystrophy was the main finding on imaging. Megaloblastic anemia was found in three cases. All the five patients were cobalamin-responsive type. Renal impairment was alleviated following treatment, edema and gross hematuria as well as hypertension disappeared later, proteinuria diminished, renal function improved, central nervous system symptoms and hematopoietic function ameliorated. CONCLUSION: In patients with hematuria, proteinuria or renal failure of unknown origin, metabolic screening and urinary organic acid analysis should be performed as early as possible to confirm the diagnosis. PMID- 16316526 TI - [Localized expressing tendency of nuclear transcription factor kappa-B, pro fibrosis genetic factors and fibronectin mRNA in renal tissues in proteinuria overload nephrotic young rats]. AB - OBJECTIVE: Terminal stage renal failure is the final common fate of chronic nephropathies independent of the type of initial insult. Abnormally filtered proteins have an intrinsic renal toxicity linked to their over-reabsorption by proximal tubular cells and activation of tubular-dependent pathways of interstitial inflammation and fibrosis. The functional importance of tubulointerstitial events in progressive renal disease is supported by evidence that the severity of tubular interstitial damage strongly correlates with the risk of renal failure. The present study aimed to investigate the expressive tendency of some pro-fibrosis genetic factors mRNA, including thrombospondin-1 (TSP-1), transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF) and the major component of extracellular matrix-fibronectin (FN) in renal tissues at different time points during early stage of renal lesions caused by proteinuria in bovine serum albumin (BSA) injection-induced proteinuria overload nephrotic young rats and the significance of these factors on tubulointerstitial fibrosis development. METHODS: Female young Wistar rats aged 3 4 weeks with proteinuria overload nephrosis induced by BSA (1.0 g/d) injected intraperitoneally were used as experimental models. The 80 young rats were divided into control group (n = 40) and BSA injected group (n = 40). At different time points (weeks 1, 2, 3 and 4), the urinary protein was measured by Coomassie brilliant blue colorimetric assay; the renal tissues morphologic changes were evaluated after HE staining; the P(65)/Rel-A, TSP-1, TGF-beta1 and CTGF mRNA expression in renal tissues was determined by in situ hybridization method and the FN mRNA expression was detected by Northern blot. The experimental data were evaluated by statistics software SPSS10.0. RESULTS: (1) Three to four weeks after BSA injection, heavy proteinuria was observed in the rats of BSA group (week 3: 104.3 +/- 21.8 mg/24 h; week 4: 131.1 +/- 18.3 mg/24 h). The proteinuria deteriorated progressively afterwards. Histopathological examination revealed that inflammatory cells infiltrated into tubulointerstitial areas extensively, protein casts were seen in tubules and edema occurred in tubulointerstitial areas. (2) In situ hybridization showed that NF-kappaB (P(65)/Rel-A) mRNA expression was up-regulated progressively in nuclei of tubular epithelial cells, the semi-quantitative scores (at 1, 2, 3 and 4 weeks) were 2.33 +/- 0.20, 2.76 +/ 0.12, 2.96 +/- 0.19, and 3.76 +/- 0.18, respectively (F = 37.34, P < 0.01). (3) At 1 week after BSA injection, the TSP-1 mRNA expression appeared in glomeruli and increased, but was light in tubulointerstitial areas, its expressive peak was observed at week 2, and declined to mild after weeks 3 and 4. The semi quantitative scores at different time points suggested that TSP-1 mRNA was expressed mainly in early stage of lesion in this model, then, this tendency turned to a flat roof smoothly. (4) TGF-beta1 and CTGF mRNA was up-regulated simultaneously in tubular epithelial cells (F = 8.80, P < 0.01 F = 19.41, P < 0.01). (5) Northern blotting showed that FN mRNA was considerably up-regulated at second week in the kidneys of rats in BSA group, 2.7-fold higher at week 4 than that at week 1 in BSA group rats, and was 3.6-fold higher than that of control group. CONCLUSION: The present study suggested that NF-kappaB (P(65)/Rel-A) mRNA expression and its activity was enhanced significantly by proteinuria-loading and synchronized with high expression of TSP-1, TGF-beta1, and CTGF mRNA in the kidney, at the same time, FN mRNA was up-regulated in renal tissues and an aggravating tendency in tubulointerstitial lesions was observed in nephrotic young rats with heavy proteinuria. PMID- 16316527 TI - [Antineutrophil cytoplasmic antibody-associated glomerulonephritis]. PMID- 16316528 TI - [Multidimensional diagnoses of attention deficit hyperactivity disorder]. PMID- 16316529 TI - [Roles of stem cells/progenitor cells in renal repair and regeneration]. PMID- 16316530 TI - [Advances of research on childhood brain development diseases--from what we know about Rett syndrome]. PMID- 16316531 TI - [Advances in molecular biological diagnosis and genotyping of group B streptococcus infection]. PMID- 16316532 TI - [Summary of sessions for pediatric nephrology, The Third World Congress of Nephrology]. PMID- 16316533 TI - [Determination of positive bronchodilating response using impulse oscillation system in children]. AB - OBJECTIVE: Impulse oscillation system (IOS) as an approach to lung function determination is suitable for children, especially for preschool children in that it only requires the individual to be examined to breathing stably with tidal volume. However, until now there have been no uniformly agreed criteria for positive bronchial reversibility test in clinical practice. A screening method was applied to seek for answer when this question in the hope of providing objective evidence for clinical diagnosis of respiratory diseases, esp. asthma. METHODS: One hundred and fifty-six children patients during asthma attack were recruited randomly for this study from January to December, 2004. These patients included 103 boys and 53 girls aged from 5 to 18 years with mean age of 8.84 +/- 2.58 years, who visited the doctors in the Clinical and Educational Centre for Asthma, Capital Institute of Pediatrics and were diagnosed as asthma. Maximal expiratory flow volume and IOS lung function were determined followed by bronchodilator reversibility test in all patients. The sensitivity and specificity of IOS parameters, total respiratory impedance (Zrs), respiratory resistance at 5 Hz (R5) and reactance at 5 Hz (X5) for diagnosing asthma at different improvement levels after inhalation of bronchodilator were calculated respectively using forced expiratory volume in first second (FEV(1)) and maximal mid-expiratory flow (MMEF) as gold standard. And the regression equation between parameters of lung function which were determined with the two different methods was analyzed. RESULTS: After inhalation of bronchodilator, the lung function of the children was improved significantly (P < 0.01), with the FEV(1), MMEF increase and Zrs, R5, and X5 decrease. There were significantly positive correlations (P < 0.01) between the improvement rate of parameters of lung functions determined with the two methods, and the highest correlation coefficients existed between X5 and FEV(1) and MMEF (respectively 0.676, 0.571), correlation coefficients between X5 and Zrs ranked second (0.519 and 0.505, respectively). When an increase of FEV(1) equal to or greater than 15%, or increase of MMEF equal to or greater than 30% was used as reference for positive bronchial reversibility, the sensitivity and specificity were relatively high for considering positive bronchial reversibility test with at least 20% decrease of Zrs, R5, and 30% decrease of X5 with IOS lung function. The sensitivity and specificity of Zrs, R5 and X5 calculated from FEV(1) were 0.62, 0.51; 0.49, 0.54; 0.70, and 0.54, respectively. The sensitivity and specificity of Zrs, R5 and X5 calculated from MMEF were 0.63, 0.54; 0.53, 0.60; 0.60, and 0.58, respectively. The regression equation showed that the decrease of Zrs, R5 and X5 corresponding to 15% increase of FEV(1) and 30% increase of MMEF were 21.7%, 21.3%; 19.9%, 19.5%; 30.1%, and 29.6%, respectively. CONCLUSION: When lung function is determined with IOS in children, only when decrease of Zrs and R5 is equal to or more than 20% and decrease of X5 is 30% or more after inhalation of bronchodilator, can the bronchial reversibility test be considered as positive. PMID- 16316534 TI - [Peroneal muscular atrophy in a case]. PMID- 16316535 TI - [Feasibility and predicted equations of spirometry in Shenzhen preschool children]. AB - Morbidity of chronic lung disease in young children is relatively high, and could increase in the future. Pulmonary function testing is used for clinical assessment of patients with suspected or obvious pulmonary disease to assess the severity of dysfunction and to evaluate therapeutic effectiveness. In the recent few years, forced expiratory parameters assessing lung function have been measured in older children. In order to assess abnormalities of lung function in preschool patients with respiratory disorders based on changes of forced expiratory parameters, adequate reference values are needed. However, such data in healthy preschool children remain scant in the literature. OBJECTIVE: The aim of this study was to characterize the spirometry of preschool children and establish the normal lung function prediction equations for Chinese preschool children. METHODS: A survey in 343 healthy preschool children (184 boys and 159 girls) aged 3 to 6 years (73 children aged 3 years, 96 children aged 4 years, 91 children aged 5 years and 83 children aged 6 years) was carried out in Shenzhen in 2004. Eleven flow volume tests parameters, i.e., forced vital capacity (FVC), forced expiratory volume at 0.5 second (FEV(0.5)), forced expiratory volume at 0.75 second (FEV(0.75)), forced expiratory volume at one second (FEV(1)), maximal mid expiratory flow rate (FEF(25%-75%)), peak expiratory flow (PEF), forced expiratory time (FET100%) were measured by using COSMED spirometry produced in Italy. Stepwise multiple regressions and non-linear regressions were carried out with the statistical software SPSS10.0 for Windows to identify the best predictors of lung function parameters using standing height, weight, age and gender as potential determinants. RESULTS: Spirometric tests could be successfully carried out by using imagery methods in the following percentages of children: 69.9% of 3 to 4 years old, 70.8% of 4 to 5 years old, 92.3% of 5 to 6 years old and 91.6% of 6 to 7 years old children, 77.7% of the selected population (217/279) of children performed at least two acceptable tests respectively. The average forced expiratory time (FET) was 1.61 +/- 0.52 sec (x +/- s), the 5th percentile value was 0.9 sec, 18 of 279 (6.5%) children produced a FET less than 1 second. Forced expiratory volume in 0.50 and 0.75 sec (FEV(0.5), FEV(0.75)) were thus measured necessary in preschool children. All lung volumes and flow rates increased with age, height as well as weight growth in both gender groups. The correlation among most lung function parameters was higher in height than in age and weight in boys. Whereas the correlation among most lung function parameters was higher in age than in height and weight in girls. The regression equations of lung function were established. By comparison with the equations derived from this study and from studies by Nystad, it was found that there was a difference between the predicted values. CONCLUSION: Spirometric testing is feasible in preschool children by using imagery methods and may be useful for both clinical practice and research. The correlation among most lung function parameters was higher in height than in age and weight in boys. Whereas the correlation among most lung function parameters was higher in age than in height and weight in girls. PMID- 16316536 TI - [Effects of immunoglobulin on the neuronal expression of IL-1beta and IL-1ra and the neuronal death at hippocampus in rats with convulsion induced by pentylenetetrazol]. AB - OBJECTIVE: To study the effects of immunoglobulin on the neuronal expression of IL-1beta and IL-1ra and the neuronal death at hippocampus in rats with convulsion induced by pentylenetetrazol. METHODS: The epilepsy model was established by injecting intraperitoneally pentylenetetrazol (PTZ) into Wistar rats. Forty-five rats were randomly divided into three groups, normal control group, PTZ plus intravenous immunoglobulin (PTZ-IVIG); PTZ plus normal saline (PTZ-NS). Neuronal death was assessed by light microscopy with the hematoxylin-eosin (HE) staining and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). IL-1beta and IL-1ra expressions were examined by histochemistry. RESULTS: The ratio of IL-1beta/IL-1ra at hippocampal CA(1) region in PTZ-IVIG group (0.5 +/- 0.1) was significantly lower than that in PTZ-NS group (1.9 +/- 0.5, t = 12.9, P < 0.05). Apoptotic cell numbers at the hippocampal CA(1) region were significantly decreased in the PTZ-IVIG group, compared to PTZ-NS group (t = 27.1, P < 0.05). The numbers of positive cells were 16.4 +/- 3.3/1000 microm(2) in the former and 41.7 +/- 3.5/1000 microm(2) in the latter. Necrotic cell numbers at the hippocampal CA(1) region were significantly decreased in the PTZ IVIG group (19.0 +/- 2.6/1000 microm(2)), compared to PTZ-NS group (42.3 +/- 4.9/1000 microm(2), t = 20.9, P < 0.05). CONCLUSION: Immunoglobulin could inhibit neuronal death induced by convulsion and its possible mechanism might be the regulation of IL-1 system in neurons. PMID- 16316537 TI - [Immunoregulatory effects of the Lonicera aquatic extract in the ovalbulmin sensitized BALB/c mice]. AB - OBJECTIVE: To evaluate the immunoregulatory effects of the Lonicera water extract in the ovalbulmin (OVA)-sensitized BALB/c mice and to explore feasibility of treating food allergy with the traditional Chinese medicine (TCM). METHODS: Forty female BALB/c mice aged 6 weeks fed with ovalbulmin-free feed, were randomly divided into 5 groups with 8 mice in each. Four groups were sensitized with OVA intraperitoneally two times and challenged intragastrically four times. Groups H, M and L were treated respectively with high (100 mg/100 ml), medium (50 mg/100ml) and low (25 mg/100 ml) concentration of the Lonicera water extract at a dose of 0.3 ml/10 g body weight just 4 hours after the first challenge and then twice daily for 10 consecutive days. The mice in group Ch were used as positive control and were sensitized intraperitoneally and treated with normal saline solution intragastrically daily. The mice in NS group were used as negative control without sensitization and challenge. Just 1 hour after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. Histological examinations on the jejunum specimens were performed after either HE or toluidine blue staining, the levels of histamine in gut of the mice were assayed with a fluorescent method; the IFN-gamma and IL-4 production in peripheral lymph node mononuclear cell (PLNMC) and the OVA-specific IgE levels in serum were measured by using ELISA; the mRNA expression of IL-12p40 in PLNMC of the mice was evaluated by RT-PCR; the footpad swelling reactions were assessed for the OVA-induced delayed hypersensitivity. RESULTS: (1) The inflammatory reactions were significantly inhibited in the mice of group H and M; the accumulated and degranulated mast cells in lamina propria were significantly reduced in the mice by gavage with 100% or 50% of the Lonicera extract, concomitant with the increased percentage of the intact mast cells. (2) The release of histamine in gut in the mice of group H and M was significantly reduced. (3) Either the IL-4 production and the ratio of IL-4/IFN-gamma in PLNMC or the IFN-gamma generation was significantly reduced in group H and M. (4) IL 12p40 mRNA expression in PLNMC was significantly reduced in group H and M. (5) The levels of OVA-specific IgE in serum were reduced in the mice of group H and M. (6) The footpad swelling reactions induced in the allergic mice were significantly inhibited after giving the Lonicera extract of the three different concentrations. CONCLUSION: The Lonicera extract showed significant immunoregulatory effects in OVA-induced allergic mice model in this study. Lonicera extract may be of potential research value in treatment of both IgE and none IgE mediated food allergy. PMID- 16316538 TI - [Construction and effect of the recombinant pshRNA plasmid against respiratory syncystial virus M2-1 gene]. AB - OBJECTIVE: Respiratory syncystial virus (RSV) is the most common cause of lower respiratory infections in infants worldwide. There is no reliable vaccine or antiviral drug against RSV at present. RNA interference (RNAi) technology is a potent method to degrade expression of the cognate mRNA. In order to inhibit the replication of RSV at gene level, the effects of specific RNAi against M2-1 gene of RSV on inhibition of viral replication in cell culture system was observed in this study. METHODS: RSV M2-1 gene, which plays a key role in RSV transcription, was chosen in this study and was used as target gene and recombinant plasmid pshRNA7816 targeting the mRNA of RSV M2-1 gene coding sequence was constructed. The pshRNA7816 was transfected into Hep2 cells. The effects of the pshRNA7816 on changes of cytopathogenic effect (CPE) of Hep2 cell induced by RSV infection were observed microscopically. Viral plaque forming assay and MTT assay were used to detect the viral titer change and protective function of the pshRNA7816 on RSV infected Hep2 cell. RESULTS: The recombinant RNAi plasmid pshRNA7816 which targets the mRNA of RSV M2-1 gene was successfully constructed. The pshRNA7816 significantly reduced CPE of RSV infected Hep2 cells, reduced the viral titer of RSV in the cells (P < 0.001). The pshRNA7816 raised the survival rate of RSV infected Hep2 cells (P < 0.001). Non-specific pshRNA plasmid did not show anti RSV effects (P > 0.05). CONCLUSION: The recombinant pshRNA7816 plasmid which targeted the mRNA of RSV M2-1 gene showed a significant and specific anti-RSV effect. PMID- 16316539 TI - [Clinical analysis of six cases with juvenile primary fibromyalgia syndrome]. AB - OBJECTIVE: To study the clinical features of juvenile primary fibromyalgia syndrome (FMS) and to evaluate outcome after treatment. METHODS: Six patients with juvenile primary FMS were registered in department of rheumatology and their clinical data were assessed, including degree of pain (visual analog scale, VAS), fatigue, depression, anxiety, sleep disturbances, arthrodynia, subjective joint swelling, abdominal pain, irritable bowel symptoms, urinary urgency, dysmenorrhea, morning stiffness, paresthesias, illness changes with weather, feeling worse with exercise, laboratory examination and outcome of treatment. RESULTS: Abdominal pain was the first symptom in 5 of the cases with juvenile primary FMS, diffuse aching and left knee pain were the first symptoms in one patient. All the 6 patients were misdiagnosed prior to their rheumatological evaluation. Diffuse aching, fatigue, sleep disturbances, illness changes with weather and feeling worse with exercise existed in all the 6 patients (100%), the mean pain score was 8.8 and the mean initial tender points (TP) count was 13.7. Arthrodynia, subjective joint swelling, abdominal pain, irritable bowel symptoms and urinary urgency existed in 5 of the 6 patients (83%). Dysmenorrhea existed in 4 (67%), depression in 3 (50%), morning stiffness in 2 (33%), paresthesias in 2 (33%) and anxiety in 2 (33%), respectively. The results of laboratory examination were normal and the outcomes of treatment were good. CONCLUSION: Juvenile primary FMS may not be a rare disease and the clinicians should pay more attention to it for avoiding misdiagnosis. PMID- 16316540 TI - [Primary IgA nephropathy in 46 children: association of clinical and pathological findings with prognosis]. PMID- 16316541 TI - [Preliminary study on rotavirus viremia in children and its clinical characteristic]. PMID- 16316542 TI - [Endoscopic variceal ligation in pediatric patients with esophageal variceal bleeding]. PMID- 16316543 TI - [Effect of topiramate on Tourette's syndrome in children]. PMID- 16316544 TI - [A case with childhood systemic lupus erythematosus with pulmonary hemorrhage as the sole initial clinical manifestation]. PMID- 16316545 TI - [A case with voltage dependent-renal tubular acidosis type IV complicated with nephrolith]. PMID- 16316546 TI - [Macrophage activation syndrome in a patient with systemic type juvenile idiopathic arthritis]. PMID- 16316547 TI - [Summary of 2005 national symposium on community acquired respiratory tract infections in children]. PMID- 16316548 TI - [Call for multi-discipline research into atherosclerotic renal artery stenosis]. PMID- 16316550 TI - [Indications for percutaneous renal angioplasty: a word of caution]. PMID- 16316552 TI - [The present status of percutaneous renal angioplasty in treatment of renal artery stenosis]. PMID- 16316553 TI - [Interventional therapy of renal artery stenosis in cardiology arena is practical and feasible]. PMID- 16316554 TI - [The choices and value of treatment strategies for atherosclerotic renal artery stenosis]. PMID- 16316555 TI - [Strengthening basic clinical skill is detrimental to integrated development of physicians]. PMID- 16316557 TI - [Changes in clinical characteristics and interventional modalities of renal artery stenosis in recent years]. AB - OBJECTIVE: To observe the changes of clinical characteristics and interventional methods of patients with renal artery stenosis in recent 17 years. METHODS: Comparing the clinical characteristics and interventional treatment of patients diagnosed renal artery stenosis by the means of renal angiography in recent 17 years. RESULTS: The numbers of renal artery stenosis were increasing year by year. the average ages before the year of 2000 was younger than that after 2000 (P < 0.05) and the main cause of renal artery stenosis was aorto-arteritis at that time, however, it turned to atherosclerosis after the year of 2000. Same patients indicated renal angiography due to plash pulmonary edema, unstable angina on abnormal serum creatinine. The interventional methods to renal artery stenosis shifted from balloon angioplasty only before the year of 2000 to stent implantation post-balloon dilatation after the year of 2000. The distal protective device was more used during the procedure. CONCLUSIONS: The numbers of patients indicated renal angiography had been increasing in the two decades. The main causes of renal artery stenosis had been considered arthro-sclerosis instead of aorta-arteritis before. The methods of treatment to renal artery stenosis had been changed to renal stenting post-balloon angioplasty after the year of 2000. PMID- 16316558 TI - [Effects of cilostazol on long-term clinical outcomes after coronary stenting]. AB - OBJECTIVE: To evaluate the effects of cilostazol on long-term clinical outcomes in patients underwent coronary stent implantation. METHODS: One hundred patients who underwent coronary stenting were randomly assigned to receive cilostazol 200 mg/d for 6 months (n = 50) or ticlopidine 500 mg/d for 1 month (n = 50). Aspirin 100 mg/d was administrated concomitantly with cilostazol or ticlopidine. Angiographic follow-up was carried out at 6 months and clinical follow-up for 3 years after stenting. RESULTS: Angiographic restenosis occurred in 5 of 34 patients (14.7%) in cilostazol group and 10 of 37 patients (27.0%) in ticlopidine group (P = 0.204). At the end of three-year follow-up, the incidence of major adverse cardiac and cerebral events (MACCE) was greatly reduced in cilostazol group compared with ticlopidine group (16% vs 36%, P = 0.023). Changes of Seattle angina questionnaire (SAQ) physical limitation score showed no significant difference between two groups (21.8 +/- 12.3 vs 16.8 +/- 15.9, P = 0.086). However, changes the improvement of angina frequency score much more was significant in cilostazol group (22.6 +/- 12.7) compared with that in ticlopidine group (16.1 +/- 13.3, P = 0.015). Recurrent angina occurred in 38% of patients in cilostazol group and 54% in ticlopidine group, respectively (P = 0.105). Readmission due to cardiac and cerebral vascular diseases was much less in cilostazol group than that in ticlopidine group (20% vs 40%, P = 0.029). CONCLUSIONS: Cilostazol treatment significantly reduced MACCE and improved the quality of life pf patients in three-year clinical follow-up after coronary stenting. PMID- 16316559 TI - [The impact of rabeprazole treatment on the quality of life in patients with reflux esophagitis and non-erosive reflux disease]. AB - OBJECTIVE: To compare the health-related quality of life (HRQL) in the patients with reflux esophagitis (RE) and non-erosive reflux disease (NERD) treated with rabeprazole in the multi-center open study. METHODS: All patients were treated with rebaprazole (10 mg, bid, ac) for eight weeks from Dec. 2002 to June 2003. 74 patients with RE; and 37 patients with NERD defined as negative endoscopic finding, the Demeester scores of 24 h pH monitoring of esophagus > 14.27 and reflux symptoms score > 6, were enrolled in. The impacts on HRQL (SF-36 questionnaire) and GERD-HRQL were assessed before and after therapy. RESULTS: At baseline, HRQL in NERD patients was impaired greater than in RE patients. After therapy, the symptoms were improved significantly in both groups. The quality of life was improved in 7 subscales in RE patients. However it was much lower in NERD patients. The scale of GERD-HRQL decreased significantly in RE patients than in NERD patients. CONCLUSIONS: NERD causes a more significant impairment in the quality of life than RE, which can be attenuated partly after 8 w rabeprazole therapy, unlike the satisfactory results favored in RE. Further research is needed to more completely understand the value of rabeprazole therapy for NERD. PMID- 16316560 TI - [The prognostic value of end-stage liver disease model in liver cirrhosis]. AB - OBJECTIVES: The prognostic ability of the model for end-stage liver disease (MELD) has been validated in many countries, but its abilities remain uncertain in China. Our aim is to evaluate the abilities of MELD in prognosis of liver cirrhosis. METHODS: A cohort of 315 patients with liver cirrhosis were retrospectively studied and followed up at least for one year. MELD score was obtained for each patient according to the modified formula by Kamath P.S.. The area under the receiver operating characteristic (ROC) curve (AUC) was used to compare MELD and Child-Turcotte-Pugh (CTP) score and classification in predicting accuracy. Kaplan-Meier survival analysis was used to compare the mortality in subgroups ranked by the MELD score. RESULTS: The AUC values generated by the ROC curves for the MELD were 0.95, 0.85 and 0.83 respectively in predicting 3, 6 month and 1 year survival, and were all more than 0.80 in predicting longer time survival, whereas the AUC was 0.82, 0.78, 0.74 for CTP score and 0.70, 0.66, 0.61 for CTP classification respectively in predicting 3, 6 month and 1 year survival. The differences of AUCs between the MELD and CTP classification were significant in predicting 6 month and 1, 3 and 4 year survival, but were not significant in predicting other time point survival. The differences of AUCs between the MELD and CTP score were not significant in predicting survival. In the subgroups of patients ranked by MELD score lower than 10, 10 to 20, 20 to 30, and more than 30, the survival rate was significant different (P = 0.000). CONCLUSIONS: MELD is a useful prognosis indicator for the liver cirrhosis. The ability of prognosis by MELD is similar to Child-Turcotte-Pugh classification and score. PMID- 16316561 TI - [The reliability of ischemic stroke subtype classification using the TOAST criteria]. AB - OBJECTIVE: The aim of this study was to assess the reliability of the classification of acute cerebral infarction based on the TOAST system. METHODS: 300 patients with ischemic stroke were recruited into this study. Ischemic stroke was classified into five subtypes according on clinical manifestations, MRI, MRA and DSA. The investigators classified all the patients into five etiologic subtypes. The data was analyzed by the statistics software of SPSS 11.5. RESULTS: The value of Kappa about the classification's reliability was 0.8, P = 0.028. CONCLUSIONS: The reliability for the classification of acute cerebral was excellent. This value could be improved with the development of the diagnosis tools and the physicians' recognition. PMID- 16316562 TI - [An analysis of multi-system involvement and renal pathology of anti-neutrophil cytoplasmic autoantibodies-related vasculitis]. AB - OBJECTIVE: To analyze the clinical and pathological characteristics of anti neutrophil cytoplasmic autoantibodies (ANCA) associated systemic vasculitis (AASV). METHODS: 426 patients with AASV diagnosed in recent 8 years in our center were retrospectively studied and their clinical and pathological data were analyzed. RESULTS: Of the 426 patients, 70 were cytoplasmic ANCA (cANCA) positive, all of them recognized proteinase 3 (PR3); 354 were perinuclear ANCA (pANCA) positive, all of them recognized myeloperoxidase (MPO). Only 47.2% (201/426) of the patients were diagnosed within three months. Clinically, 87.1% (371/426) had renal involvement and 68.0% (83/122) had crescentic glomerulonephritis. 61.0% (260/426) had lung involvement. Other clinical manifestations included gastrointestinal symptoms (43.0%), nervous system disorder (15.7%), ophthalmic (19.5%), otologic (23.2%) and nasal involvement (8.2%). The prevalences of fatigue and renal involvement were significantly higher in patients with pANCA than those in patients with cANCA (63.8% vs 47.1%, P < 0.01; 88.7% vs 78.6%, P < 0.05, respectively). The prevalences of ophthalmic, nasal involvement, skin rash and arthralgia were significantly higher in patients with cANCA than those in patients with pANCA (31.4% vs 16.9%, P < 0.01; 21.4% vs 5.6, P < 0.01; 28.6% vs 16.4%, P < 0.05; 44.3% vs 28.0%, P < 0.01, respectively). The patients were treated with corticosteroid and cyclophosphamide. The remission rate in induction phase was 88.5%. CONCLUSIONS: AASV is a common autoimmune disease in Chinese. For patients with multi-system damage, an ANCA test should be performed in order to make an early diagnosis and start therapy in time. PMID- 16316563 TI - [The pulmonary expression of granulocyte-macrophage colony-stimulating factor and surfactant protein in adult idiopathic pulmonary alveolar proteinosis]. AB - OBJECTIVE: To investigate the expression of granulocyte-macrophage colony stimulating factor (GM-CSF), GM-CSF/IL-3/IL-5 receptor common beta chain (GM CSFbetacR), surfactant protein (SP)-A and SP-D in adult patients with idiopathic pulmonary alveolar proteinosis (PAP), so as to demonstrate the possible association of GM-CSF, GM-CSFbetacR, SP-A and SP-D with human PAP. METHODS: The protein expression of GM-CSF, GM-CSFbetacR, SP-A and SP-D in the alveolar macrophages and alveolar type II epithelial cells was detected by immunohistochemical staining. RESULTS: The age was 51.8 +/- 9.5 years old for patients with idiopathic PAP (6; males 4, females 2) and 48.8 +/- 12.6 years old for the control adult individuals (6; males 4, females 2). There was no difference in the expression of GM-CSF (P = 0.818) and SP-D (P = 0.24) in alveolar macrophages between patients with idiopathic PAP and the controls. The expression of GM-CSFbetacR (P = 0.002) and SP-A (P = 0.004) in alveolar macrophages of patients with idiopathic PAP was less than that in the controls. There was no expression of GM-CSF in alveolar type II epithelial cells of patients with idiopathic PAP and the controls. There was a little expression of GM-CSFbetacR in alveolar type II epithelial cells of the controls. There was no expression of GM-CSFbetacR in alveolar type II epithelial cells of patients with idiopathic PAP. There was no difference in the expression of SP-A (P = 0.818) and SP-D (P = 0.485) in alveolar type II epithelial cells between patients with idiopathic PAP and the controls. CONCLUSION: The decreased protein expression of GM-CSFbetacR in alveolar macrophages may be one of the mechanisms causing dysfunction of the macrophages in adult patients with idiopathic PAP. PMID- 16316564 TI - [The application of gonadotropin in treatment of male central hypogonadism]. AB - OBJECTIVE: To observe the efficacy of human chorionic gonadotrophin (hCG) and hCG plus human menopausal gonadotropin (HMG) for central hypogonadism in male patients. METHODS: 64 men with central hypogonadism were recruited in this study, including 19 patients with Kallmann syndrome, 41 patients with idiopathic hypogonadotrophic hypogonadism (IHH) and 4 patients with hypogonadism after brain surgery. 33 patients were treated with hCG 1500 IU intramuscularly twice a week, whereas 31 patients were treated with intramuscular hCG 1500 IU plus HMG 75 IU twice a week, for at least 6 months. RESULTS: After treatment, all patients felt stronger physically and 42/64 patients developed beard, pubes or armpit hair. The testis volume enlarged significantly [(3.08 +/- 2.44) ml vs (8.92 +/- 5.37) ml, P < 0.001], and serum follicle-stimulating hormone, luteinizing hormone and testosterone concentrations were higher significantly than those before treatment (P < 0.05). 6/64 patients underwent spermatorrhea and 2 patient were found to have spermatogenesis. If judged by the testis volume, 52 patients (81.2%) were effective and 12 patients were ineffective. CONCLUSIONS: For male patients with the central hypogonadism, hCG and hCG plus HMG can promote the pubertal development and maturation of second sex characteristics, as well as enhance the physical strength; in some patients both androgen production and spermatogenesis can be achieved. PMID- 16316566 TI - [The clinical characteristics of primary intracranial germinoma]. AB - OBJECTIVE: To analyse the clinical characteristics of intracranial germinoma. METHODS: Retrospective analysis was applied to study the clinical characteristics of 26 intracranial germ cell tumor patients admitted to our hospital during 1991 2003. The clinical, biochemical and imaging profiles including human chorionic gonadotropin-beta, alphafetoprotein, MRI and CT as well as treatments were analysed. RESULTS: 26 intracranial germ cell tumor patients were admitted to our hospital during 1991-2003, accounting for 0.9% of all intracranial tumors (3020 cases) at the same time. Among these patients 19 cases (73.1%) were primary intracranial germinoma, 9 patients (47.4%) were female and 10 patients (52.6%) were male. 13 patients (68.4%) were younger than 20 years. 14 patients (73.7%) had headache, vomiting and nausea, 8 patients (42.1%) had diabetes insipidus, 5 patients (26.3%) had hypopituitarism. 9 patients' tumors (47.3%) were in pine region, 7 patients' tumors (38.8%) were in sellar region. 14 patients (73.7%) were treated wit radiotherapy and all of them were discharged with good condition. 10 patients were treated with operation (7 patients accepted radiotherapy after operation) and 2 of them died after operation. CONCLUSIONS: Intracranial germinoma mainly affects female children and adolescents, pine and suprasellar regions are the commonly involved regions, the most common manifestations of intracranial germinoma are headache, vomiting, nausea, diabetes insipidus and hypopituitarism. Radiotherapy has good efficacy in the treatment of intracranial germinoma. PMID- 16316567 TI - [A randomized controlled and multicenter clinical study of ferrous L-threonate in treatment of iron deficiency anemia]. AB - OBJECTIVE: To study the clinical efficacy and side effects of ferrous L-threonate for treatment of iron deficiency anemia (IDA). METHODS: It is a multicentral, randomized, double blind, double placebo and paralled comparative study with positive control. One hundred and forty IDA patients diagnosed according to the standard criteria in three hospitals were randomly divided into a test group (ferrous L-threonate plus placebo ferrous succinate) and a positive control group (ferrous succinate plus placebo ferrous L-threonate). Some iron parameters were examined 1, 4 and 8 weeks after medication. Hemoglobin, reticulocyte and other parameters for safety observation were collected every two weeks. RESULTS: For the 2 groups, self comparison showed significant difference (P < 0.01). The total efficacy is 98.44% and 97.01% respectively with no difference. Hemoglobin rised rapidly and gradually and reached a peak in week 8, the change was statistically significant (P < 0.01). Changes of iron parameters also showed significant difference. Side-effects were similar in both groups (13.85% and 14.71%, P > 0.05). CONCLUSION: The effect of ferrous L-threonate in IDA treatment is significant and rapid. Side-effects are few and minimal. PMID- 16316569 TI - [The role of CD25 antibody in unrelated hematopoietic stem cell transplantation]. AB - OBJECTIVE: To explore the role of CD(25) antibody on engraftment and graft-versus host disease (GVHD) prophylaxis in unrelated hematopoietic stem cell transplantation (UHSCT). METHODS: CD(25) 1 mg/kg was given on day 1, day 4 post transplantation in 27 patients of UHSCT. RESULTS: Hematopoietic recovery was obtained in 26 patients. One patient died before hematopoietic recovery. Acute GVHD occurred in 17 patients and 6 patients with II or more than degree acute GVHD (23%). Three patients experienced relapse and other 3 patients with serious infection. In these 26 eligible patients, 19 patients got disease free survival. CONCLUSIONS: CD(25) antibody plays an important role on engraftment and GVHD prophylaxis in the treatment of UHSCT and does not increase rate of leukemia relapse. It provides a way of GVHD prophylaxis in unrelated and HLA mismatched hematopoietic stem cell transplantation. PMID- 16316588 TI - The role of inflammation as a pathogenic factor in the development of renal disease in diabetes. PMID- 16316590 TI - Newer agents for the treatment of painful diabetic peripheral neuropathy. AB - Neuropathic pain is responsible for a significant amount of the morbidity associated with generalized and focal peripheral neuropathies in diabetes. It is a consequence of alterations in neuronal function, chemistry, and structure that occur secondary to nerve injury. A variety of agents from diverse pharmacologic classes, the so-called adjuvant analgesics, have been used to treat neuropathic pain. These include antidepressants, first- and second-generation anticonvulsants, antiarrhythmic agents, topical agents, N-methyl-d-aspartate receptor antagonists, and the opioid analgesics. The availability of several newer agents, used alone or in combination, has resulted in the successful alleviation of neuropathic pain in many patients. Recent advances in the understanding of pain mechanisms at multiple central nervous system levels should pave the way toward more effective treatment modalities with less prominent side effects. PMID- 16316591 TI - Recent developments in the assessment of efficacy in clinical trials of diabetic neuropathy. AB - A large number of measures may be employed in clinical practice and for epidemiologic studies to quantify and risk stratify diabetic patients with neuropathy. However, not all measures are suitable for assessing the benefits of therapeutic intervention. Therefore, for the purpose of this review we focus on measures that may be employed to define the efficacy of interventions in clinical trials of human diabetic neuropathy. Two major types of end points are used: 1) those that assess symptoms for defining efficacy in painful diabetic neuropathy, and 2) those that assess neurologic deficits that assess the effects of treatments that may prevent further degeneration or promote repair. PMID- 16316592 TI - The pivotal role of offloading in the management of neuropathic foot ulceration. AB - Lower extremity amputations among persons with diabetes are generally preceded by neuropathic foot ulcerations. Healing of diabetic ulcerations in a timely manner is of central importance in any plan for amputation prevention. With sufficient vascular supply, appropriate debridement, and infection control, the primary mode of healing a diabetic neuropathic foot ulcer is pressure dispersion. The total contact cast has been deemed by many to be the gold standard in offloading; however, modification of a standard removable cast walker to ensure patient compliance may be as efficacious in healing diabetic foot ulcers as the total contact cast. Combining an effective, easy to use offloading device that ensures patient compliance with advanced wound healing modalities may form a formidable team in healing ulcers and potentially averting lower limb amputations. PMID- 16316593 TI - Theories concerning the pathogenesis of the acute charcot foot suggest future therapy. AB - The acute Charcot foot is characterized by bone fracture and dislocation, and is a rare complication of distal symmetrical neuropathy in diabetes. The cause is unknown, but it cannot be attributed solely to loss of protective sensation. However, recent advances in understanding the mechanisms of osteoclast activation have suggested that the key abnormality may lie in an enhanced inflammatory response to injury, which is itself linked to increased bone lysis. The recognition that the acute Charcot foot is essentially an inflammatory arthropathy suggests new options for the management of this potentially devastating condition. PMID- 16316594 TI - 2005: The International Diabetes Federation focuses on the diabetic foot. AB - The International Diabetes Federation (IDF) has proclaimed 2005 to be the "Year of the Diabetic Foot." Together with the International Working Group on the Diabetic Foot, the IDF has launched a year-long campaign to raise awareness of the diabetic foot worldwide. In this article, both organizations are briefly described, an overview of the aims of the global awareness campaign is given, and the activities to promote World Diabetes Day in 2005 are outlined. Attention is also paid to present and future developments to improve the standards of diabetic foot care worldwide. PMID- 16316596 TI - Lipids and diabetic renal disease. AB - Diabetic nephropathy is commonly associated with dyslipidemia, but the role of lipids in the progression of this disorder remains unresolved. In particular, the role of lipid-lowering drugs, such as 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors and fibrates, as renoprotective agents is not clarified. Experimental studies have demonstrated that dietary lipids promote renal injury and that statins, independent of their lipid-lowering effects, confer renoprotection via effects on intrarenal hemodynamics and renal cytokine and chemokine expression. Clinical studies have in general been underpowered, but a recent meta-analysis and findings from the Heart Protection Study suggest that statins may be renoprotective. Nevertheless, with the convincing antiatherosclerotic effects of these agents, including in the setting of diabetes, they should be widely administered in the diabetic population with or at risk for nephropathy. PMID- 16316597 TI - Pathogenic role of nitric oxide alterations in diabetic nephropathy. AB - Diabetic nephropathy is the most frequent cause of terminal renal failure, requiring renal replacement therapy. Although a number of factors may contribute to the development of renal disease in diabetes, the recent past has witnessed an explosive growth in literature pertaining to the role of nitric oxide in diabetic nephropathy. However, there are significant controversies in the findings of these studies partly because of the complex metabolic pathways involved in the generation and fate of nitric oxide in the diabetic kidney. The following discussion presents a critical and balanced review of the current understanding of this subject. PMID- 16316598 TI - Early nephropathy in type 1 diabetes: a new perspective on who will and who will not progress. AB - Impaired renal function and end-stage renal disease (ESRD) affect up to a third of patients with type 1 diabetes. Thus, strategies for early detection and for preventative interventions are of critical importance. A model of diabetic nephropathy was developed in the 1980s that placed paramount importance on the finding of microalbuminuria as an early marker of a committed process of progressive kidney disease in diabetes. However, recent studies have provided evidence that microalbuminuria is a marker of dynamic, rather than fixed, kidney injury. Preliminary studies into early renal function decline, a process measured in early nephropathy using a simple assay for cystatin C to calculate the slope of glomerular filtration rate change over time, suggest that it is a more proximal marker than microalbuminuria of a person's trajectory toward impaired renal function and ESRD. Therefore, early renal function decline, rather than microalbuminuria, may be considered as the early marker of the committed process underlying progressive diabetic nephropathy. PMID- 16316599 TI - Proteomics and diabetic nephropathy. AB - Diabetes mellitus is acknowledged to be a group of metabolic diseases and heterogeneous in natural history, pathogenesis, response to treatment, and disease progression and remission. Diabetic nephropathy (DN) accounts for approximately 40% of all newly diagnosed cases of end-stage renal disease. The complexity of diabetes and its complications requires a broad-based, unbiased, scientific approach such as proteomics. Recently, proteomics (the systematic analysis of protein identity, quantity, and function) has been applied to the study of DN. Proteomic investigations into diabetic kidney disease have identified new mechanisms of diabetic renal pathology, as well as potential urinary markers of DN. Other current proteomic advances in understanding DN include identifying the role of advanced glycation end products in decreased mitochondrial respiration and also the rapid development of mass spectrometric methods for protein and peptide markers of DN development and markers to pharmacologic therapies. Proteomic analysis has only recently been applied to the study of DN, yet it has shown substantial potential. PMID- 16316600 TI - PPAR-gamma agonists and diabetic nephropathy. AB - Diabetic nephropathy is a clinical syndrome of albuminuria, declining glomerular filtration rate, and increased risk of cardiovascular disease. Multiple mechanisms have been implicated in its pathogenesis. Although current therapies appear to be effective, treatment of diabetic nephropathy remains suboptimal. This review summarizes the recently emerging evidence suggesting that peroxisome proliferator-activated receptor-gamma agonists may prove to be effective therapeutic agents in the treatment of diabetic renal complications. PMID- 16316601 TI - [Seven-year clinical experience with the SVL/Beznoska implant for total knee arthroplasty]. AB - PURPOSE OF THE STUDY: The aim of the study was to evaluate our seven-year experience with the anatomic, non-constrained SVL/Beznoska implant for total knee replacement. MATERIAL: A total of 374 cemented total knee replacements, using the SVL/Beznoska implant, were evaluated in 333 patients treated at the First Orthopedic Clinic Charles University in Prague, during a seven-year period. In addition, a patellar replacement was used in five of these patients. Included in the evaluation were also eight non-cemented and hybrid SVL/Beznoska implants used 6 years ago. METHODS: The aim of the study was to evaluate the results in terms of implant function. Therefore, the longevity of implants was assessed by Kaplan Meier's survival analysis. Another criterion was the flexion achieved. To be able to compare our clinical results involving the SVL/Beznoska implant with other, foreign types of implants, the patients were evaluated on the basis of the New Jersey Orthopaedics Hospital Knee Evaluation System. RESULTS: The Kaplan-Meier's survival score was evaluated. The mean maximum flexion achieved in the patients with SVL implants was 107 degrees . The evaluation by the New Jersey Orthopaedics Hospital Knee Evaluation System showed excellent outcomes in 269 (72 %) patients, good outcomes in 79 (21 %) patients, satisfactory in 19 (5 %) and poor in 7 (2 %) patients. There was no mechanical failure of the implant, such as break of an implant or dislocation of a PE component. Six infected implants and four aseptic loosenings were recorded. Three patients diagnosed with an infected total knee replacement had been treated for rheumatoid arthritis for a long time. One patient was on dialysis for renal insufficiency and one had erysipelas. DISCUSSION: The results of our group with the SVL/Beznoska implant were compared with those obtained from the group of 63 patients who underwent total knee arthroplasty with the LCS implant. In these patients, the mean maximum flexion achieved was 105 degrees , and the criteria of the Knee Evaluation System showed excellent outcomes in 81 %, good in 16 % and satisfactory in 3 % of the patients. No unsatisfactory outcome was recorded. Considering the fact that the LCS implant was used only in younger and active patients, the outcomes achieved with the SVL/Beznoska system can be regarded as very good. Of the six patients with infectious complications, three were treated with immunosuppressive therapy for rheumatoid arthritis, one was on dialysis for renal insufficiency and one experienced a flare-up of erysipelas. CONCLUSIONS: The most important outcome of the evaluation of 374 total knee replacements, using the SVL implant carried out at the First Orthopedic Department of the First Faculty of Medicine, Charles University, was the finding of a perfect function of all components in all patients treated. None of the complications occurring in our group was associated with the use of this implant. PMID- 16316602 TI - [Pathologic proximal femoral fractures in children in an unicameral bone cyst]. AB - PURPOSE OF THE STUDY: Proximal femoral fractures in children are rare, pathologic fractures being extremely rare. Despite many meanings these fractures are still "unsolved" there are some definite rules for treatment of true accidental injuries. Pathologic fractures are outstanding with their extremely rare incidence. The aim of the study is to overview a large clinical material, find out the incidence of this pathologic fracture, the extent and shape of the unicameral bone cyst (UBC), specific therapeutic approach, technical problems of eventual osteosynthesis, number of reoperations and sequels. MATERIAL: Altogether 49 children with 50 accidental and pathologic fractures of proximal part of the femur treated in the Regional Pediatric Trauma Centre of the Department of Pediatric and Trauma Surgery, 3rd Faculty of Medicine, Charles University, Prague. METHODS: Retrospective and prospective study of children (0 to 15 years of age) treated with proximal femoral accidental and pathologic fractures during the 20 year period (from August 1984 to November 2004). Classification of fractures according to Delbet and Colonna. Diagnosis of bone cyst with plain X rays, eventually CT scans. RESULTS: During the 20 years period (August 1984 to November 2004) 49 children with 50 proximal femoral fractures were treated in the Department. Four patients sustained a pathologic fracture through an unicameral bone cyst. Two of these latter children were treated by an open reduction and osteosynthesis with the use of the proximal femoral AO-ASIF angled-plate and two children nonoperatively using skeletal traction because of impossibility of insertion of the osteosynthetic material without a damage of the growth plate. Subsequent operations of the UBC were necessary in these two children. All four patients recovered well without sequels. DISCUSSION: Pathologic fractures in UBC are usually treated nonoperatively and the cyst itself is treated after fracture healing. Proximal femoral impairment is the exception from this rule because of weigh bearing necessity. However, there may be problems with insertion of the implant when the cyst is very near to capital physis and traction treatment is then the method of choice with a delay of operative treatment of the cyst. CONCLUSIONS: Pathologic proximal femoral fractures in UBC are extremely rare and need individual approach. Some of them should be operated on the others primarily treated by traction with secondary operation of the cyst. Complications can be frequent. PMID- 16316603 TI - [Resection hip arthroplasty--mid- and long-term results]. AB - PURPOSE OF THE STUDY: The study deals with resection hip arthroplasty, which is currently one of the principal orthopedic surgery procedures.The aim of the study was to evaluate the mid- and long-term results of resection arthroplasty carried out for different indications and to ascertain effects of the extent of proximal femur resection on the functional outcome. MATERIAL: In the period from 1979 to 2000, we carried out 159 resection hip arthroplasties for septic and aseptic loosening of total hip arthroplasty (THA), septic arthritis or osteomyelitis. Of these only the patients who underwent no THA implantation or reimplantation within 2 years of the operation were evaluated. We assessed 102 patients (71 women and 31 men) with 105 resection arthroplasties for the following indications: 14 cases of aseptic loosening of the THA, 72 cases of septic loosening and 19 cases of hip inflammation. The average follow-up was 70.8 months (range, 26 months to 20 years). The average age of the patients at the time of surgery was 62.5 years (29 to 86 years). METHODS: We recorded the patient's subjective evaluation of the surgery outcome, shortening of the extremity, and the range of hip motion. Harris hip scores were used to assess the functional outcome. We also evaluated, and statistically analysed, the extent of resection of the proximal femur on X-ray images and its effect on the eventual shortening of the extremity; this was done on the basis of the Grauer classification system. RESULTS: The average Harris hip score was 63.2 points (range, 47 to 88). None of the joints achieved excellent outcomes; 8.5 % were good, 22 % were satisfactory and 73 % were poor. On pain evaluation, 35 % of the patients were completely free from pain, 18 % reported mild pain not interfering with daily activities, 27 % experienced moderate pain responding to common analgetics and 11 % had severe pain necessary to be treated with strong analgetics. Persistent rest pain not responding to analgetics was in 9 % of the patients. The average shortening of the extremity for the whole group was 4.1 cm (range, 3.5- 7 cm): the average values for shortening in type I, type II and type II resections were 3.25 cm (3-5 cm), 4.1 cm (3-6 cm) and 5.3 cm (4.5/7 cm), respectively. The range of passive flexion was on average 81 degrees (range, 45-105 degrees ). Limping and a positive Trendelenburg's test were recorded in all patients and all also had to use walking aids. The outcome of surgery was evaluated as satisfactory by 35 % and as partially satisfactory by 43 % of the patients. DISCUSSION: Resection arthroplasty is a reliable technique for pain reduction, but it results in a considerable alteration of hip function. Pain in the hip after surgery is bearable, as 80 % of our patients reported only mild or moderate pain alleviated by common analgetics, or complete absence of pain. This is in agreement with the results of Stoklasa (84 %) and Stedry (79 %). However, the subjective evaluation of surgery by patients is different; Petty has reported 14 % of satisfied patients, while Ahlgren and Bohler described 100 % of satisfied patients. Hip stability and less shortening of the extremity are more important for patients' satisfaction than a certain mild degree of pain. The best subjective evaluation was reported by the patients who had resection at the femoral neck level and who also had the lowest average shortening of the extremity. But only 55 % of the satisfied or partially satisfied patients reported mild or no pain. CONCLUSIONS: Resection arthroplasty is, in indicated cases, a reliable method which, in spite of certain drawbacks, produces good functional outcomes and makes the patients satisfied. It is, without doubt, an integral part of the current orthopedic surgery repertoire. We recommend a cautious approach to repeated attempts at THA reimplantation or to reconstruction procedures. PMID- 16316604 TI - [Allograft for surgical reconstruction of the cruciate ligaments of the knee - part 1]. AB - PURPOSE OF THE STUDY: The growing numbers of anterior cruciate ligament (ACL) reconstructions performed by an increasing number of surgeons have resulted in an increased number of failed reconstructed ligaments. For repeat surgery, autologous tissues are most frequently used, namely BTB graft, hamstring tendons or quadriceps tendon construct. However, these alternative methods have certain disadvantages and therefore we decided to use BTB allografts from cadaverous donors. The risk of disease transmission due to allograft implantation has been reported to be low, but a thorough serological screening of donors is the prerequisite. We used BTB allografts first in revision ACL surgery only, but because of good results we started using theme for reconstruction of both cruciate ligaments and, in some cases, also for primary reconstruction. MATERIAL: In the period from 2002 to 2004, patellar ligaments harvested from 23 cadaverous donors were used at the orthopedic ward of the Ceske Budejovice hospital to prepare 87 BTB grafts, of which 42 were implanted. In 57 % of the procedures, an allograft was used in revision surgery carried out for the failure of a ;previously reconstructed ACL in 10 % it was used in complete reconstruction of both cruciate ligaments, and in 14 % it was used for primary ACL reconstruction in indicated cases. In 19 % of the cases, allograft was used when autologous graft failed or was damaged during the primary operation. METHODS: Graft harvesting, storage, handling and implantation have been carried out in accordance with the practices included in Act no. 285/2002 Coll. Serological examination of the donors and bacteriological assays of the grafts were performed according to the current regulations. The implantation of BTB allografts in ACL reconstruction was carried out by the standard method used in reconstructive procedures. RESULTS: Out of 87 BTB allografts prepared, 16 were discarded because of positive culture findings. Two allografts could not be used because the screening was positive for CMV infection. No infectious complications, poor healing or a systemic response to the allograft implanted were recorded in any of the patients. DISCUSSION: The use of allografts is a method suitable for revision surgery in failed ACL reconstruction. It allows us to avoid further weakening of the structures associated with the knee joint. The size of allograft bony blocks permits treatment of defects in the tibial or the femoral tunnel. CONCLUSIONS: The use of a BTB allograft is a reliable and safe procedure. It has advantages not only in revision ACL reconstruction, but also in reconstruction of both cruciate ligaments. Also primary ACL reconstruction in indicated cases achieved good results. PMID- 16316605 TI - [BTB allograft for revision surgery of the anterior cruciate ligament - part 2]. AB - PURPOSE OF THE STUDY: The study evaluates our experience with revision anterior cruciate ligament (ACL) surgery, with emphasis placed on the use of allografts. MATERIAL: In the 2002-2004 period, 421 reconstructions of the anterior cruciate ligament, using patellar BTB or hamstring tendon autografts, were carried out in our orthopedic ward. In the same period we performed 24 revision ACL reconstructions (6 %) with BTB allografts; 19 were in men and five in women. Revision surgery after ACL reconstruction with a BTB graft fixed with a non absorbable interference screw and with a hamstring tendon graft was performed in 18 and 5 patients, respectively. METHODS: We distinguish four steps in the revision procedure: 1) diagnosis and analysis of the ACL reconstruction failure; 2) preoperative planning and surgery timing (one- or two-stage procedure, graft type, fixation method); 3) operative procedure; 4) postoperative care including rehabilitation. Revision surgery is indicated on the basis of subjective complaints (instability), and the results of clinical examination (Lachman's and pivot-shift tests) and imaging methods (X-ray, MRI, arthroscopy). Causes of failure are categorized as 1) traumatic (major trauma, too early weight-bearing, minor trauma due to rehabilitation); 2) surgery-related (erroneous position of the tibial and/or femoral tunnels, insufficient tensioning of the graft and its insufficient fixation); 3) biological (poor ;graft incorporation and restructuring, infection); and 4) combination of all previous causes. Errors in tunnel position are differentiated according to the part of the tunnel (tibial, femoral or both) tunnel direction (ventral, dorsal, lateral or medial) and the degree of malposition (mild, moderate or serious). We perform one-stage surgery when the position of tunnels is correct, with the exception of revision due to infection, and in all malpositions but for a dorsally positioned tibial tunnel. In a moderate degree of femoral tunnel ventral malposition, we make decisions individually. A two-stage procedure consists of removal of the failed graft and fixation material and spongioplasty followed by revision surgery. The results of revision reconstruction greatly depend on a correct isometric position. We extend the tunnel, if it is in a mild-degree malposition, and create a new, smaller tunnel, if the malposition is severe. Fixation, with either the Rigidfix system or interference screws, is also selected according to the direction and degree of malposition. In the last 3 years, we used exclusively grafts harvested from cadavers. RESULTS: In the group of 24 patients undergoing revision ACL reconstruction, the right knee was treated in 13. The previous ACL reconstruction was done with BTB grafts in 18 patients, with hamstring tendons in 5 patients and one patient underwent reconstruction by Harnach's method in an outside institution. The average time between the primary reconstruction and revision surgery was 27 months (range, 4 to 169 months). We found a surgery-associated error in 12 cases. poor graft restructuring in 3 and involvement of traumatic etiology in 11 cases. One patient underwent revision surgery because of infection. We used one-stage procedures in 20 patients and two-stage procedures in four patients. We fixed the graft with femoral interference screws and the Rigidfix system in 17 and 7 patients, respectively, and with tibial interference screws in 23 patients (absorbable screw completed with cancellous screw in one patient). Only in one patient did we use the tibial Rigidfix system. The average follow-up was 16 months. No infection, thromboembolic disease or synovialitis were recorded. One patient experienced a recurrent failure of the graft and one patient was treated for the Cyclops lesion. The average Lysholm scores were 78.25 (range, 48-97); 87.5 % of the patients were satisfied with the results and the same proportion of patients would undergo the surgery again. The overall results appeared poorer due to the patients in whom revision ACL reconstruction was performed on arthritic joints. DISCUSSION: The outcomes of revision surgery are worse than those in primary reconstruction. We regard allografts with massive bony blocks, adjusted as required, as an optimal method. The risk of disease transmission is low, operative time is shorter, incision is smaller and further trauma to the treated or the other, healthy knee due to graft harvest is avoided. Only patients without signs of gonarthrosis who have motivation are indicated for revision surgery, because they can be expected to cooperate well in the postoperative period. CONCLUSIONS: Revision ACL surgery should be performed by surgical teams with sufficient experience in this field. The crucial point is the analysis of ACL reconstruction failure with further procedure planning. The use of BTB allografts from the local tissue bank proved efficient in our hospital. In the hands of experienced surgeons, allograft offer great prospects for ACL reconstruction with good outcome. PMID- 16316606 TI - [Correlation of long-term clinical and radiological results after meniscectomies]. AB - PURPOSE OF THE STUDY: Although surgical treatment of meniscus injuries has made great progress, meniscectomy remains the most frequently indicated intervention. The aim of this retrospective study was to evaluate the long-term radiological results of isolated subtotal and total meniscectomies and compare them with the clinical results. MATERIAL: The assessed group included 22 men and 8 women at an average age of 35 years; nine and 21 knees, respectively, were treated by isolated subtotal and total meniscectomy in the period from 1987 to 1989. The lateral meniscus was operated on in six knees and the medial meniscus in 24 knees. Patients with other injuries to the knee joint or with a systemic disease were not evaluated. The average follow-up was 16 years. The results were compared with the other, non-treated knee. METHODS: The results were assessed by means of the Lysholm scoring system which takes into account limping, support requirement, walking distances, instability, edema, stair ascent and descent, ability to squat and pain. Radiographic osteoarthritis of the knee was classified by the Kellgren and Lawrence system. A grade greater or equal to 2 was taken for a clear sign of arthritis. In addition, the anatomical axis of the lower extremity was evaluated. Anteroposterior projection of both knees under load was made in a standing position on long films and lateral projection was obtained separately on short films. RESULTS: The average Lysholm scores were 87 points (range, 41-100) and 91 points (range, 67-100) in the treated and non-treated knees, respectively. The outcome of meniscectomy was excellent in 14 (47 %), good in eight (27 %), satisfactory in five (16 %) and poor in three (10 %) knees. In 12 patients (40 %) the state of the treated knee was evaluated as being equal to that of the non treated knee and in eight patients (27 %) it was even better. Osteoarthritis of the affected compartment was found in eight treated knees and that of the corresponding compartment on the opposite side in four knees. Grade 2 Kellgren- Lawrence radiographic changes were present in four, grade 3 in two and grade 4 in two knees treated by meniscectomy. The anatomic femoro-tibial angle differed between the treated and non-treated side, by 4 degrees on average (range, 2 degrees to 10 degrees ), in 13 (43 %) patients. DISCUSSION: Johnson et al. found at follow-up for about 17 years that osteoarthritis developed more often in the treated than in healthy contralateral knees (40 % as against 6 %). Tapper and Hoover reported that 45 % of men and 10 % of women were without subjective complaints at 10 to 30 years of follow up. In the relevant Czech literature, only Pasa et al. have recently paid attention to the development of lesions in knees treated by meniscectomy, but they have largely evaluated changes occurring after partial arthroscopic meniscectomies. CONCLUSIONS: The present study showed that osteoarthritis after subtotal or total meniscectomy developed in 27 % of the patients at an average follow-up of 16 years. In less than half of the patients, a deviation of the anatomical axis occurred in the treated lower extremity, as compared with the other healthy side. Lysholm scores worse in the treated than the non-treated knee were found in one third of the patients.However, a deviated knee joint axis and/or osteoarthritic lesions did not always correlate with deteriorated clinical findings. PMID- 16316607 TI - [Opening-wedge osteotomy of the proximal tibia]. AB - PURPOSE OF THE STUDY: The aim of this study was to evaluate the accuracy of correction and short-term outcomes of treatment by opening-wedge tibial osteotomy with the use of allograft and a modified Puddu plate fixation. MATERIAL: Between February 2001 and March 2003, 33 operations on 30 patients, at an average age of 54 years, with gonarthrosis and varus deformity were evaluated. Grade III arthritis was diagnosed in 24 knees (73 %) and grade II in nine knees (27 %). METHODS: The preoperative planning as well as intraoperative correction were based on the limb mechanical axis, as shown on an X-ray image of the whole lower extemity. Tibial osteotomy was performed from the medial approach and the position was fixed, in contrast to the original method, with an allograft and a T plate. The corrrection was planned so that the mechanical limb axis should intersect the articular space at 62 % of its length laterally. The limb was postoperatively immobilized for 4 weeks in a knee brace allowing walking. Limited weight bearing was recommended from the third week. The outcome of treatment was evaluated at 11 months of follow-up by the Lysholm scoring system. RESULTS: Correction of the limb mechanical axis involved the axis intersecting the plane of the joint space on average at 60.2 % of its length, as against the planned 62 %; the average deviation from the planned correction was 2.1 degrees. Lysholm scores improved on average by 23 points. Improvement by 20 points was regarded as a good outcome and was achieved in 31 joints (94 %). Two joints (6 %) showed poor outcomes due to associated complications. The complications included a fissure of the lateral tibial plateau in one knee, failed stabilization with subsequent loss of correction and pseudoarthrosis development in one joint, prolonged healing with a partial loss of correction (5 degrees) in one patient, and superficial infection in one patient. DISCUSSION: In patients with knee arthritis and varus deformity, the achievement of optimal correction of the mechanical limb axis is the prerequisite for a long-term good outcome. Optimal correction, as characterized by Puddu, is attained when the mechanical axis intersects the joint space at 62 % of its length laterally. Hernigou et al. define the hip-knee-ankle angle in the range of 183 degrees -186 degrees as an optimal postoperation correction. In our experience, osteotomy combined with our modification of fixation is an exact method for correcting the axis; in addition, it is simple and is associated with a low occurrence of postoperative complications. In view of a possible total knee arthroplasty, there is less disturbance in the tibial plateau-diaphysis alignment and the tibial tuberosity level is preserved. CONCLUSIONS: Opening-wedge tibial osteotomy with the use of a Puddu plate and allograft in our modification is an effective, exact and simple method of correcting the lower extremity axis, and is associated with few complications. We do not recommend this method for interventions planned to achive corrections of more than 16 degrees, because these have a higher occurrence of complications. The use of allograft seems to have good prospects; another option for achieving large correction includes application of an LCP plate. PMID- 16316608 TI - [Juvenile unstable severe slip of the proximal femoral epiphysis: case report and review of treatment options]. AB - The authors present the case of a severe unstable slip of the proximal femoral epiphysis in a 14-year-old boy. A serious metabolic disorder diagnosed in the boy led to postponement of necessary surgical intervention. The primary development of necrosis of the proximal femoral epiphysis during the 6 months following the slip is reported here. A good outcome was eventually achieved by three-plane valgus, flexion and derotational osteotomy of the femur at second-stage surgery. The authors emphasize the prognostic importance of stability assessment in the displaced epiphysis. PMID- 16316609 TI - [Unilateral dislocations of the thoracic and lumbar spine]. AB - Unilateral dislocation is a trauma typical of the cervical spine. Case reports on unilateral dislocation of the thoracic or lumbar vertebrae can be found in the relevant literature only rarely and they often describe this condition associated with multiple trauma or combined injuries. Although unilateral dislocation is an unstable injury with rotation involved, injury to the spinal cord or spinal nerve roots is not common. Diagnosis is based on radiographic and CT examination. Therapy includes open reduction and instrumented spondylodesis of the injured segment. The cases of two patients with unilateral dislocations in the thoracic and the lumbar spine, respectively, treated at the Department of Spinal Surgery, Motol Teaching Hospital in Prague, are reported here. PMID- 16316610 TI - If only it were simple. PMID- 16316611 TI - Chronic myelogenous leukemia. PMID- 16316612 TI - Resistance to imatinib: mechanisms and management. AB - Imatinib, a specific small molecule inhibitor of the Abl kinase, has become the standard drug therapy for chronic myelogenous leukemia in all phases. More than 80% of newly diagnosed patients with chronic phase attain a complete cytogenetic response (CCR). Although remissions in patients with early disease are generally durable, acquired resistance after an initial response is common in advanced disease. Reactivation of Bcr-Abl signaling is almost invariably present at the time of relapse, consistent with re-establishment of the initial pathogenetic mechanism. Mutations in the kinase domain (KD) of Bcr-Abl that impair drug binding and increased expression of Bcr-Abl have been identified as major mechanism of acquired drug resistance. The fact that Bcr-Abl remains central to disease pathogenesis at the time of relapse implies that it also remains the optimal drug target. Alternative Abl kinase inhibitors with increased potency and activity against most Bcr-Abl KD mutants are currently undergoing phase I/II clinical testing, with encouraging early results. Despite the high rates of CCR, persistence of residual leukemia as assessed by reverse transcription polymerase chain reaction is the rule even in patients with chronic phase, suggesting that even these patients may remain at risk of relapse. Understanding the mechanisms underlying disease persistence will be crucial for developing strategies to eradicate residual leukemia. PMID- 16316613 TI - Cancer- and treatment-related anemia. PMID- 16316614 TI - Parenteral iron therapy: a single institution's experience over a 5-year period. AB - Many patients require parenteral iron therapy for optimal correction of anemia, including cancer patients who require erythropoietic drugs. Available parenteral iron therapy options include iron dextran, iron gluconate, and iron sucrose. The purpose of this study is to summarize our institution's experience with parenteral iron therapy over a 5-year period, with a focus on comparative safety profiles. All patients receiving parenteral iron therapy over this period were included in the analysis. Chi-squared test and Fisher's exact test were used to compare the adverse event rates of each product. A total of 121 patients received 444 infusions of parenteral iron over this period. Iron dextran was the most commonly used product (85 patients) and iron sucrose was the least used (2 patients). Iron gluconate was used by 34 patients. Overall adverse event rates per patient with iron dextran and iron gluconate were 16.5% and 5.8%, respectively (P = .024). Premedication with diphenhydramine and acetaminophen before infusions of iron dextran reduced adverse event rates per infusion from 12.3% to 4.4% (P = .054). Test doses of iron dextran were used 88% of the time for initial infusions of iron dextran. All adverse events for all parenteral iron products were mild or moderate. There were no serious adverse events and no anaphylaxis was observed. Our results suggest that, if test doses and premedications are used, iron dextran is an acceptable product to treat iron deficiency. PMID- 16316615 TI - Cancer patient survival and erythropoietin. AB - Cancer patients are frequently anemic. Treatment of anemic patients with erythropoiesis-stimulating proteins (ESPs) such as epoetin and darbepoetin is associated with benefits that include a reduced transfusion risk and improved quality of life. The recent reports of two randomized trials in which ESP treatment was associated with a decreased survival raised valid concerns regarding the safety of these agents in oncology practice. Reports of erythropoietin receptors on non-hematologic human tumor cells have increased the level of concern and provided a relatively simple model for the effects of ESPs on tumor progression and resistance to treatment. This article reviews available data, which lead to a number of conclusions: 1) the two trials suggesting a negative impact on survival have serious methodologic issues that may compromise interpretation; 2) when used to treat rather than prevent anemia in cancer patients, ESPs show no significant negative impact on survival outcomes; 3) with the exception of erythroleukemia cell lines, the presence of functional erythropoietin receptors on human tumor cells has not been conclusively shown; and 4) a sound theoretical basis exists, supported by preclinical evidence, that any effect of ESP therapy on tumor outcomes may depend on baseline hemoglobin levels, with different effects when anemic and non-anemic individuals are treated. For the present, it is prudent to withhold ESP therapy unless hemoglobin concentrations fall below 12 g/dL and to titrate treatment to maintain a target of 12 g/dL, with adjustments in therapy to insure that levels do not exceed 13 g/dL. PMID- 16316616 TI - Benefits associated with an early hemoglobin response to epoetin alfa therapy in the treatment of chemotherapy-related anemia. AB - Although previous studies have recognized that timely correction of anemia is desirable, no published research quantifies the association between the timeliness of the hemoglobin rise and patients' outcomes. This study evaluates whether anemic patients with cancer who are receiving chemotherapy and who experienced an early response to epoetin alfa (> or = 1 g/dL hemoglobin increase at the end of 4 weeks of treatment) experienced better clinical and drug utilization outcomes compared with patients who did not experience an early response. Three large, open-label, community studies of epoetin alfa for the treatment of chemotherapy-related anemia were retrospectively analyzed to assess the association of early hemoglobin response to subsequent transfusion requirements, subsequent hemoglobin response, quality of life, and epoetin alfa dosage administered over the study. Two epoetin alfa dosing regimens were evaluated: 10,000 units 3 times weekly with potential escalation to 20,000 units, and 40,000 units once weekly with potential escalation to 60,000 units. In all studies, patients who experienced an early hemoglobin response had statistically lower subsequent transfusion requirements, higher rates of subsequent hemoglobin response, shorter time to hemoglobin response, better improvements in quality of life scores, and lower average weekly epoetin alfa dose than patients who did not experience an early hemoglobin response. Similar proportions of patients experienced early response in the 3-times weekly and once-weekly epoetin alfa regimens. This ad hoc analysis found that early hemoglobin response to epoetin alfa therapy was associated with improved clinical benefits and drug utilization. Early hemoglobin response may therefore be considered as a desired goal of epoetin alfa therapies. PMID- 16316617 TI - Assessment of mosquito larvicidal potency of cyclic lipopeptides produced by Bacillus subtilis strains. AB - In this study, mosquito larvicidal potency of cyclic lipopeptides (CLPs) secreted by two Bacillus subtilis strains were determined. LC50 of the crude CLPs secreted by B. subtilis DM-03 and DM-04 strains against third instar larvae of Culex quinquefasciatus was 120.0+/-5.0 and 300.0+/-8.0mg/l respectively post 24 h of treatment. Physico-chemical factors such as pH of water, incubation temperature, heating and exposure to sunlight hardly influenced the larvicidal potency of these CLPs. Present study provided the evidence that B. subtilis lipopeptides were safe to Indian major carp Labeo rohita, a non-target aquatic organism. These properties of B. subtilis CLPs can be exploited for the formulation of a safer, novel biopesticide for effective control of mosquito larvae. PMID- 16316618 TI - A novel protein refolding method using a zeolite. AB - We have succeeded in developing a simple and effective protein refolding method using the inorganic catalyst, beta-zeolite. The method involves the adsorption of proteins solubilized with 6M guanidine hydrochloride from inclusion body (IB) preparations onto the zeolite. The denaturant is then removed, and the proteins in the IBs are released from the zeolite with polyoxyethylene detergent and salt. All of the IBs tested (11 different species) were successfully refolded under these conditions. The refolded proteins are biochemically active, and NMR analysis of one of the proteins (replication protein A 8) supports the conclusion that correct refolding does occur. Based on these results, we discuss the refolding mechanism. PMID- 16316619 TI - Identification of heavy metal complexes of a hexapeptide inhibitor of the human immunodeficiency virus integrase protein by using a voltammetric approach. AB - Complexation of the hexapeptide Hys-Cys-Lys-Phe-Trp-Trp, inhibitor of the human immunodeficiency virus integrase protein, with the heavy metal ions Cd2+, Pb2+, and Zn2+ has been investigated using differential pulse polarography. In the case of Pb2+, no significant complexation is detected, whereas in the cases of Cd2+ and Zn2+, strong and electrochemically inert ML2 complexes predominate. In contrast, ML complexes are present in a low proportion or are absent. When possible, the corresponding conditional stability constants have been determined at both pH 7.0 and pH 7.5, showing that Zn2+ complexes are slightly more stable than Cd2+ complexes. PMID- 16316620 TI - A complete set of hyaluronan fragments obtained from hydrolysis catalyzed by hyaluronidase: Application to studies of hyaluronan mass distribution by simple HPLC devices. AB - Hyaluronan (HA) has different biological functions according to its molar mass; short HA fragments are involved in inflammation processes and angiogenesis, whereas native HA is not. Physicochemically, studies of native HA hydrolysis catalyzed by bovine testicular hyaluronidase (HAase) have suggested that kinetic parameters depend on HA chain length. To study the influence of HA chain length in more detail, and to try to correlate the physicochemical and biological properties of HA, HA hydrolysis catalyzed by HAase was used in a new procedure to obtain HA fragments of different molar masses. HA fragments (10-mg scale) with a molar mass from 800 to 300,000 g mol(-1) were prepared, purified using low pressure size exclusion chromatography (SEC), lyophilized, and characterized in molar mass by either mass spectrometry or HPLC-SEC-multiangle laser light scattering. The polydispersity index of the purified fractions was less than 1.25. The complete set of HA standards obtained was used to calibrate our routine HPLC-SEC device using only a refractive index (RI) detector. We showed that the N acetyl-d-glucosamine reducing end assay and the calibrated HPLC-SEC-RI gave equivalent kinetic data. In addition, the HPLC-SEC-RI furnished the mass distribution of the polysaccharide during its hydrolysis. PMID- 16316621 TI - Measurement of electric current evoked by substrate transport via bi-directional H+/oligopeptide transporter over-expressed in HeLa cells: electrogenic efflux and existence of a newly observed channel-like state. AB - In the present study, we measured an electric current induced by substrate transport in a HeLa cell over-expressing a human intestinal di/tri-peptide transporter using the whole-cell patch-clamp technique. Gly-Sar, a typical substrate, induced an inward current associated with its uptake, which showed concentration-dependency following Michaelis-Menten-type kinetics with an apparent K(0.5) of 1.3mM as well as voltage-dependency. An outward current accompanying the efflux of Gly-Sar was also observed after washing out the cell. This outward current was voltage-dependent and was reduced by the inward proton gradient. In the case of hydrophobic dipeptides such as Gly-Phe and Gly-Leu, a distinctive current was observed: after washing out the cells, no outward current was observed, but rather, an 'inward leak' current was sustained in spite of the absence of transportable substrate. This leaky current was abolished by the perfusion of Gly-Sar and subsequent washing. It is considered that the hydrophobic substrate sticks within the substrate-binding site and causes the newly observed state, or the 'inward leak' current. PMID- 16316622 TI - Sustained phenotypic reversion of junctional epidermolysis bullosa dog keratinocytes: Establishment of an immunocompetent animal model for cutaneous gene therapy. AB - Gene transfer represents the unique therapeutic issue for a number of inherited skin disorders including junctional epidermolysis bullosa (JEB), an untreatable genodermatose caused by mutations in the adhesion ligand laminin 5 (alpha3beta3gamma2) that is secreted in the extracellular matrix by the epidermal basal keratinocytes. Because gene therapy protocols require validation in animal models, we have phenotypically reverted by oncoretroviral transfer of the curative gene the keratinocytes isolated from dogs with a spontaneous form of JEB associated with a genetic mutation in the alpha3 chain of laminin 5. We show that the transduced dog JEB keratinocytes: (1) display a sustained secretion of laminin 5 in the extracellular matrix; (2) recover the adhesion, proliferation, and clonogenic capacity of wild-type keratinocytes; (3) generate fully differentiated stratified epithelia that after grafting on immunocompromised mice produce phenotypically normal skin and sustain permanent expression of the transgene. We validate an animal model that appears particularly suitable to demonstrate feasibility, efficacy, and safety of genetic therapeutic strategies for cutaneous disorders before undertaking human clinical trials. PMID- 16316623 TI - Lovastatin-induced RhoA modulation and its effect on senescence in prostate cancer cells. AB - Lovastatin inhibits a 3-hydroxy 3-methylglutaryl coenzyme A reductase and prevents the synthesis of cholesterol precursors, such as farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), responsible for important cell signaling in cell proliferation and migration. Recently, the anti-cancer effect of lovastatin has been suggested in various tumor types. In this study, we showed that a low dose lovastatin induced senescence and G1 cell cycle arrest in human prostate cancer cells. Addition of GGPP or mevalonate, but not FPP, prevented the lovastatin-induced G1 phase cell cycle arrest and cell senescence. We found that constitutively active RhoA (caRhoA) reversed lovastatin-induced senescence in caRhoA-transfected PC-3 cells. Thus, we postulate that modulation of RhoA may be critical in lovastatin-induced senescence in PC-3 cells. PMID- 16316624 TI - Interaction of myocilin with the C-terminal region of hevin. AB - Myocilin, a matricellular protein, is mutated in glaucoma. Here we report the identification and characterization, by the yeast two-hybrid system, of a putative interacting protein with myocilin. One of the positive clones exhibited 100% identity with the carboxyl-terminal (C-t) region of hevin, a member of the BM-40/SPARC/osteonectin family of extracellular matrix proteins. Protein interaction was assayed, in doubly transfected 293-T cells, by Western blot and fluorescent microscopy. Western blot analysis of the culture medium and lysates from cotransfected cells indicated that myocilin causes intracellular accumulation of hevin-C-t and impairs its secretion. This effect on hevin-C-t was augmented when coexpressed with the myocilin P370L mutant, known to cause a severe form of glaucoma. By fluorescent microscopy, myocilin localizes with hevin C-t in the Golgi in cotransfected 293-T cells and with hevin-wt in the ocular ciliary epithelium. Overall, these results suggested that the C-t of hevin contains important determinants for interaction with myocilin. PMID- 16316625 TI - Depletion of CPAP by RNAi disrupts centrosome integrity and induces multipolar spindles. AB - We previously identified a novel microtubule-destabilizing motif in CPAP that can disassemble microtubules. To examine further the CPAP function in human cells, we used siRNA to knockdown its expression. Our results showed that CPAP depletion arrested cells in mitosis and induced apoptosis. Interestingly, more than 40% of these mitotic cells had multiple spindle poles. Furthermore, inhibition of the kinesin Eg5 in CPAP-depleted cells resulted in monopolar spindles, indicating that Eg5 function is required for multipolar spindle formation in the absence of CPAP. Together, our results reveal a structural role for CPAP to maintain centrosome integrity and normal spindle morphology during cell division. PMID- 16316626 TI - Translocation of positively charged copoly(Lys/Tyr) across phospholipid membranes. AB - Much attention has recently been paid to the study of positively charged polypeptides as a possible carrier for therapeutic protein or DNA delivery to cells. In this study, we have investigated the translocation of positively charged copoly(Lys/Tyr) (MW=72000, DP=385) across lipid membranes constituted from egg-phosphatidylcholine (EPC), dioleoyl-phosphatidylethanolamine (DOPE), as well as soybean phospholipids (SBPL) using zeta potential method, circular dichroism spectroscopy (CD), electrophysiology technique, fluorescence spectroscopy, and confocal laser scanning microscopy. Results of zeta potentials show that copoly(Lys/Tyr) associate with lipid membranes and become gradually saturated on the membranes either hydrophobically or electrostatically or both. CD studies demonstrate that the copoly(Lys/Tyr) takes and remains beta-sheet conformation during its interaction with liposome membranes, indicating that the translocation process should be carpet-mode like. Data from the electrophysiology technique reveal that positively charged copoly(Lys/Tyr) can cause transmembrane currents under an applied voltage, confirming its transfer across lipid membranes. Fluorescence spectroscopy results display a three-step mechanism of translocation across membrane: adsorption, transportation, and desorption, which has been verified by results from confocal laser scanning microscopy. We provided the first direct observation that the positively charged polypeptides, copoly(Lys/Tyr), can translocate through SBPL and EPC/DOPE lipid bilayer membranes. In addition, we found that the translocation efficiency of copoly(Lys/Tyr) was higher on the EPC/DOPE lipid membrane than on the SBPL lipid membrane. PMID- 16316627 TI - Oncogenic protein UnpEL/Usp4 deubiquitinates Ro52 by its isopeptidase activity. AB - UnpEL (also known as Usp4 or Unph) is an oncogenic protein, because its expression with a strong promoter results in the tumorigenic transformation of NIH3T3 cells injected into nude mice. Although the structure of UnpEL is that of a deubiquitinating enzyme, neither its precise function in mammalian cells nor the mechanism of UnpEL-mediated tumorigenesis is known. Here, we show that UnpEL functions as a deubiquitinating enzyme in human HEK293T cells and its isopeptidase activity deconjugates ubiquitin specifically from a UnpEL interacting protein Ro52. We further show that UnpEL translocates to the cytoplasmic rod-like structures and colocalizes with Ro52 when Ro52 is overexpressed in HEK293 cells. These results suggest that UnpEL colocalizes with the unubiquitinated form of Ro52 to the cytoplasmic rod-like structures, where it keeps Ro52 unubiquitinated. The continuous deubiquitination of Ro52 might be involved in tumorigenesis. PMID- 16316628 TI - p16INK4a and p14ARF methylation as a potential biomarker for human bladder cancer. AB - Promoter hypermethylation is one of the putative mechanisms underlying the inactivation of negative cell-cycle regulators. We examined whether the methylation status of p16(INK4a) and p14(ARF), genes located upstream of the RB and p53 pathway, is a useful biomarker for the staging, clinical outcome, and prognosis of human bladder cancer. Using methylation-specific PCR (MSP), we examined the methylation status of p16(INK4a) and p14(ARF) in 64 samples from 45 bladder cancer patients (34 males, 11 females). In 19 patients with recurrent bladder cancer, we examined paired tissue samples from their primary and recurrent tumors. The methylation status of representative samples was confirmed by bisulfite DNA sequencing analysis. The median follow-up duration was 34.3 months (range 27.0-100.1 months). The methylation rate for p16(INK4a) and p14(ARF) was 17.8% and 31.1%, respectively, in the 45 patients. The incidence of p16(INKa) and p14(ARF) methylation was significantly higher in patients with invasive (>or=pT2) than superficial bladder cancer (pT1) (p=0.006 and p=0.001, respectively). No MSP bands for p16(INK4a) and p14(ARF) were detected in the 8 patients with superficial, non-recurrent tumors. In 19 patients with tumor recurrence, the p16(INK4a) and p14(ARF) methylation status of the primary and recurrent tumors was similar. Of the 22 patients who had undergone cystectomy, 8 (36.4%) manifested p16(INKa) methylation; p16(INK4a) was not methylated in 23 patients without cystectomy (p=0.002). Kaplan-Meier analysis revealed that patients with p14(ARF) methylation had a significantly poorer prognosis than those without (p=0.029). This is the first study indicating that MSP analysis of p16(INK4a) and p14(ARF) genes is a useful biomarker for the pathological stage, clinical outcome, and prognosis of patients with bladder cancer. PMID- 16316629 TI - Roles of distinct cysteine residues in S-nitrosylation and dimerization of DJ-1. AB - A significant proportion of early onset parkinsonism is inherited as an autosomal recessive trait (AR-EP). DJ-1 was identified as one of the causative genes for AR EP (PARK7), and DJ-1 protein has been implicated in oxidative stress response through oxidation of one of the three cysteine residues (i.e., Cys106). However, the individual roles of these cysteine residues remained unclear. We show by a systematic mutagenesis analysis that Cys46 and Cys53 of DJ-1, but not Cys106, are susceptible to S-nitrosylation in vitro as well as in cultured cells. Furthermore, alanine substitution of Cys46 diminished dimerization of DJ-1, a fundamental feature of this protein. These results indicate that distinct cysteine residues of DJ-1 harbor differential roles in relation to its structure and function. PMID- 16316630 TI - Evaluation of viral and mammalian promoters for driving transgene expression in mouse liver. AB - Fifteen luciferase plasmid constructs driven by various promoters including cytomegalovirus (CMV), Rous sarcoma virus (RSV), human serum albumin (SA), alpha 1 antitrypsin (AAT), cytochrome P450 CYP1A2, CYP2C9, CYP2C18, CYP2D6, CYP3A4, mouse CYP2b10, human amyloid precursor protein (APP), chicken beta actin (ACT), nuclear factor kappa B (NFkappaB), and heat shock protein 70 (HS) promoters were hydrodynamically introduced into mouse hepatocytes, and the level and persistence of luciferase gene expression were examined. Eight hours post-gene transfer, the CMV and AAT promoters showed the highest activity, followed by the CYP2D6, HS, and RSV promoters which were slightly less active. The human serum albumin promoter exhibited the lowest activity among the promoters examined. The time course of gene expression showed a two-phase decline in luciferase activity with a rapid phase within the first 5-7 days and a slower decline thereafter. Results from Southern and Northern blot analyses revealed a good correlation between the decline of luciferase activity and the decrease in mRNA level, suggesting promoter silencing as the possible mechanism for the observed transient luciferase gene expression. Inclusion of EBN1 and oriP sequences of Epstein-Barr virus into the plasmid extended the period of active transcription for about one week. These results provide important information concerning the role of promoters in regulating transgene expression and for the proper design of plasmids for gene expression and gene therapy. PMID- 16316631 TI - High-fat diet feeding impairs both the expression and activity of AMPKa in rats' skeletal muscle. AB - OBJECTIVE: To investigate the effects of high-fat feeding on the expression and activity of AMPK in rats' skeletal muscle. METHODS: Total 40 male Wistar rats were randomly divided into three groups and received either a rat maintenance diet (Control group) or an isocaloric rich-fat diet (HF group and MET group) for five months. Metformin was administered orally with the daily dose of 300mg in MET group during the last month of high-fat feeding. Hyperinsulinemic-euglycemic clamp study was performed to estimate whole-body insulin sensitivity. The ability of insulin-stimulated glucose uptake in isolated skeletal muscle was detected just before execution. mRNA levels of AMPKa1, AMPKa2, and Glut4 of rats' skeletal muscle were determined using real-time PCR. Protein contents of AMPKa, P-AMPKa, P ACC, and Glut4 in rats' skeletal muscle were measured using Western blot. RESULTS: (1) Hyperinsulinemic-euglycemic clamp study revealed a significantly impaired insulin action at the whole-body level after high-fat feeding (p<0.01). Also, both basal and insulin-stimulated glucose uptake in isolated skeletal muscle decreased after high-fat feeding (p<0.05), indicating onset of high-fat induced insulin resistance. (2) Five months of high-fat treatment induced a significant decrease of AMPKa protein contents and AMPKa2 mRNA levels in rats' skeletal muscles (p<0.05), while it did not alter AMPKa1 mRNA levels. Protein levels of P-AMPKa also decreased after high-fat feeding (p<0.01). These data suggest that high-fat exposure might impair AMPKa expression and activities. (3) P-ACC protein contents, mRNA and protein levels of Glut4 in rats' skeletal muscles also decreased after high-fat treatment (p<0.05). (4) Compared with HF group, although no significant alternations of AMPKa expression in rats' skeletal muscles were detected, P-AMPKa levels revealed a 162% increase after metformin treatment (p<0.05), demonstrating the AMPK-activating effect of metformin. Accompanied with activation of AMPKa, rats in MET group exhibited significantly elevated P-ACC contents, Glut4 mRNA and protein levels, and an obviously enhanced insulin sensitivity at both whole-body and skeletal muscle levels (p<0.05). CONCLUSIONS: High-fat feeding impaired both the expression and activities of AMPKa, while activating AMPKa by metformin obviously ameliorated high-fat induced insulin resistance, thus indicating a possible role of AMPKa in lipotoxicity. PMID- 16316632 TI - Characterization of multimetric variants of ubiquitin carboxyl-terminal hydrolase L1 in water by small-angle neutron scattering. AB - Here, we illustrated that the morphological structures of ubiquitin carboxyl terminal hydrolase L1 (UCH-L1) variants and Parkinson's disease (PD) exhibit good pathological correlation by a small-angle neutron scattering (SANS). UCH-L1 is a neuro-specific multiple functional enzyme, deubiquitinating, ubiquityl ligase, and also involved in stabilization of mono-ubiquitin. To examine the relationship between multiple functions of UCH-L1 and the configuration of its variants [wild type, I93M (linked to familial Parkinson's disease), and S18Y (linked to reduced risk of Parkinson's disease)], in this report, we proposed that these were all self-assembled dimers by an application of a rotating ellipsoidal model; the configurations of these dimers were quite different. The wild-type was a rotating ellipsoidal. The globular form of the monomeric component deformed by the I93M mutation. Conversely, the S18Y polymorphism promoted the globularity. Thus, the multiple functional balance is closely linked to the intermolecular interactions between the UCH-L1 monomer and the final dimeric configuration. PMID- 16316633 TI - The role of pharmacodynamic research in the assessment and development of new antibacterial drugs. AB - Antibacterial resistance continues to increase world wide, with some bacterial pathogens exhibiting resistance to virtually all available drugs. As the plague of antibacterial resistance continues to grow and create serious therapeutic problems, it is essential that the development of new antibacterial agents continue. Pharmacodynamic research plays an important role in the development of new antibacterial agents, as pharmacodynamic data can help define the clinical potential of a new drug and identify the strengths and weaknesses in comparison to other drugs already on the market. Furthermore, pharmacodynamic experiments can help focus the clinical phases of drug development by providing key information on the pharmacodynamic parameters that influence efficacy and the pharmacodynamic targets that should be achieved to optimize clinical success. Characterization of these pharmacodynamic properties for a new drug in development can help direct the design of the best dose and dosing strategy for clinical trials. This review will focus on the tools, methods, and strategies used to characterize the pharmacodynamics of antibacterial agents and aide in their development for clinical use. PMID- 16316634 TI - Enhancing the antiproliferative effect of topoisomerase II inhibitors using a polypeptide inhibitor of c-Myc. AB - Topoisomerase II inhibitors are widely used in cancer chemotherapy. However, their use is limited by severe adverse effects to normal tissues, including cardiotoxicity. One approach to reduce the cytotoxicity in normal tissues may be to sensitize cancer cells to the toxicity of these agents, allowing them to be administered in a lower and safer dose. A hallmark of many types of cancer is overexpression of c-Myc, and a molecule which targets c-Myc will affect the cancer cells more significantly than the normal tissues. This report demonstrates that pretreatment of cells with a polypeptide, which inhibits c-Myc transcriptional function causes cells to be more susceptible to the topoisomerase II inhibitors doxorubicin and etoposide. Inhibition of c-Myc and Max dimerization by this polypeptide leads to as much as a 2-fold reduction in the doxorubicin and etoposide IC(50) in three different cell lines tested. Furthermore, the c-Myc inhibitor affects the cell cycle distribution of MCF-7 breast cancer cells by enhancing the G(0)/G(1) accumulation induced by doxorubicin and etoposide. We have shown that this effect is not due to enhanced drug accumulation or inhibited drug efflux. Rather, it is likely due to the transcriptional consequences of c Myc inhibition, specifically reduction in the levels of the polyamine synthesizing enzyme ornithine decarboxylase. In summary, our results suggest that polypeptides, which inhibit c-Myc transcriptional function, may prove to be a useful tool in combination therapy with topoisomerase II inhibiting drugs. PMID- 16316635 TI - Serotonin induces pulmonary artery smooth muscle cell migration. AB - The chronic phase of pulmonary arterial hypertension (PAH) is associated with vascular remodeling, especially thickening of the smooth muscle layer of large pulmonary arteries and muscularization of small pulmonary vessels, which normally have no associated smooth muscle. Serotonin (5-hydroxytryptamine, 5-HT) has been shown to induce proliferation and hypertrophy of pulmonary artery smooth muscle cells (PASMC), and may be important for in vivo pulmonary vascular remodeling. Here, we show that 5-HT stimulates migration of pulmonary artery PASMC. Treatment with 5-HT for 16h increased migration of PASMC up to four-fold as monitored in a modified Boyden chamber assay. Increased migratory responses were associated with cellular morphological changes and reorganization of the actin cytoskeleton. 5-HT induced alterations in morphology were previously shown in our laboratory to require cAMP [Lee SL, Fanburg BL. Serotonin produces a configurational change of cultured smooth muscle cells that is associated with elevation of intracellular cAMP. J Cell Phys 1992;150(2):396-405], and the 5-HT4 receptor was pharmacologically determined to be the primary activator of cAMP in bovine PASMC [Becker BN, Gettys TW, Middleton JP, Olsen CL, Albers FJ, Lee SL, et al. 8 Hydroxy-2-(di-n-propylamino)tetralin-responsive 5-hydroxytryptamine4-like receptor expressed in bovine pulmonary artery smooth muscle cells. Mol Pharmacol 1992;42(5):817-25]. We examined the role of the 5-HT4 receptor and cAMP in 5-HT induced bovine PASMC migration. PASMC express 5-HT4 receptor mRNA, and a 5-HT4 receptor antagonist and a cAMP antagonist completely blocked 5-HT-induced cellular migration. Consistent with our previous report that a cAMP-dependent Cl( ) channel is required for 5-HT-induced morphological changes in PASMC, phenylanthranilic acid, a Cl(-) channel blocker, inhibited actin cytoskeletal reorganization and migration produced by 5-HT. We conclude that 5-HT stimulates PASMC migration and associated cytoskeletal reorganization through the 5-HT4 receptor and cAMP activation of a chloride channel. PMID- 16316636 TI - Increased number of aldosterone-sensitive NTS neurons in Dahl salt-sensitive rats. AB - Dahl salt-sensitive rats develop severe hypertension during a high-sodium diet, but the basis of their salt-sensitive phenotype is not completely understood. A subset of neurons in the nucleus tractus solitarius (NTS) are uniquely sensitive to the adrenal steroid hormone aldosterone, which is critically involved in sodium homeostasis, due to their expression of the enzyme 11-beta-hydroxysteroid dehydrogenase type 2 (HSD2). The number of HSD2 neurons in the NTS was counted in prehypertensive 7-week-old Dahl salt-sensitive rats and compared with two control strains: Dahl salt-resistant and Sprague-Dawley rats. Dahl salt-sensitive rats had more HSD2 neurons than age-matched Dahl salt-resistant and Sprague-Dawley rats (24% and 21%, respectively). Cell counts were also made in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats; the number of HSD2 neurons in both of these strains was similar to the values obtained for Sprague-Dawley rats. The increased number of HSD2-immunoreactive neurons counted in Dahl salt sensitive rats suggests that they may have a greater number of aldosterone sensitive NTS neurons. Alternatively, an increase in HSD2 expression in Dahl salt sensitive rats could increase the overall immunoreactivity, permitting detection of more of these neurons. In either case, the roughly 20% increase in HSD2 neurons in the NTS of prehypertensive Dahl salt-sensitive rats is a novel factor associated with their salt-sensitive phenotype. These neurons may play a role in regulating sodium appetite, which is abnormally suppressed in Dahl salt-sensitive rats. PMID- 16316637 TI - Changes in reflex responses of the genioglossus muscle during sleep in rabbits. AB - Changes in reflex responses in the genioglossus (GG) muscle evoked by electrical stimulation of the inferior alveolar nerve and GG muscle tone (background activity, BGA) were investigated during sleep-wakefulness stages in rabbits. The GG muscle showed two types of electromyographic activity patterns: a respiration related phasic activity and non-respiration-related activity. GG reflex responses and BGA exhibited a stage-dependent decrease as they were constantly suppressed from quiet wakefulness to non-rapid eye movement sleep to rapid eye movement sleep (REMS). Degree of suppression of reflexes was much larger than that of BGA regardless of GG activity patterns. When amplitude of reflex responses was compared between with and without rapid eye movements during REMS, no difference between the conditions was noted. These results suggest that excitability of the GG muscle is affected by sleep stages by not only a modulation of excitability in motoneurons but also in interneurons involved in the reflex arc. PMID- 16316638 TI - Human cytomegalovirus inhibits Akt-mediated eNOS activation through upregulating PTEN (phosphatase and tensin homolog deleted on chromosome 10). AB - OBJECTIVES: Atherosclerosis is the leading cause of death in the United States, and human cytomegalovirus (HCMV) infection may play a role in the development of this disease. Diminished expression and/or activity of endothelial nitric oxide synthase (eNOS) are an early event in atherogenesis. In the current study, we investigated the effects of HCMV infection on eNOS activation in human aortic endothelial cells (HAECs). METHODS AND RESULTS: We found that HCMV inhibited eNOS phosphorylation/activation in HAECs. The signaling upstream of eNOS involving Akt and PDK1 were also suppressed by the HCMV infection. Moreover, HCMV infection increased the expression of PTEN (phosphatase and tensin homolog deleted on chromosome 10). Silencing PTEN expression with specific siRNA reversed the inhibitory effects on eNOS activation in HCMV-infected cells indicating the involvement of PTEN in mediating HCMV's inhibitory effects. Next we observed that the activation of p38 MAPK stress signaling pathway mediates HCMV's effects on PTEN up-regulation and eNOS inactivation. CONCLUSIONS: In summary, our findings suggest that inhibition of eNOS leading to endothelial dysfunction may be a basis of the pro-atherogenic effects of HCMV. Importantly, upregulation of PTEN and activation of stress signal p38 MAPK are involved in HCMV's inhibitory effects on eNOS activation. PMID- 16316639 TI - Polymorphisms of glutathione S-transferase M1, T1, and P1 in patients with HBV related liver cirrhosis, chronic hepatitis, and normal carriers. AB - OBJECTIVES: Persistent hepatitis B virus (HBV) infection often leads to the development of chronic hepatitis and cirrhosis. The role of host genetic factors in chronic HBV infection is not fully understood. We studied the influence of glutathione S-transferase (GST) M1, T1, and P1 polymorphisms in patients with different stages of HBV infections. METHODS: The sample population included 41 HBV normal carriers, 37 patients with chronic hepatitis, and 38 patients with cirrhosis (infected with HBV) compared to a control group (n = 59). PCR-based procedures were performed in the studied populations to confirm the genotypes of GSTT1, M1, and P1. Odds ratio analysis tests were used for statistical evaluation. RESULTS: We found that the frequency of GSTP1-Val (105)/Val (105) genotype was significantly higher in patients with liver cirrhosis (27%) than HBV normal carriers (2.4%; OR 14.8, 95% CI 1.8-122.5) and the frequency GSTP1-Val (105)/Ile (105) genotype was significantly higher in patients with liver cirrhosis (59.5%) than HBV normal carriers (19.5%; OR 6.1, 95% CI 2.1-16.7). The genotype GSTP1-Val (105)/Val (105) was more frequent in patients with chronic hepatitis (19.4%) than HBV normal carriers (2.4%; OR 9.65, 95% CI 1.1-82.8). Patients with cirrhosis also had a higher frequency of the GSTM1 null genotype (71.1%) than HBV normal carriers (27.5%; OR 6.5, 95% CI 2.4-17.4) and the GSTM1 null genotype was more frequent in patients with chronic hepatitis (64.9%) than HBV normal carriers (27.5%OR 4.9, 95% CI 1.8-12.8). The frequency of GSTT1 genotype was similar in all groups. CONCLUSION: These results suggest that in HBV infection, inheritance of the null GSTM1 and GSTP1-Val (105) polymorphisms involves a host genetic factor that is relevant to disease progression. PMID- 16316640 TI - Quantification of urinary allantoin by capillary zone electrophoresis during recombinant urate oxydase (rasburicase) therapy. AB - OBJECTIVES: Rasburicase (Fasturtec) is used to prevent or treat hyperuricemia associated with chemotherapy. We developed a capillary zone electrophoresis method to measure urinary allantoin, the degradation product of uric acid by rasburicase. DESIGN AND METHODS: Electrophoresis was performed using a P/ACE 5500 system (Beckman) with a fused silica capillary tube and a UV-visible detector set at 214 nm. Urine samples from 10 patients with non-Hodgkin's lymphoma were analyzed to validate the technique. RESULTS: Using a sodium tetraborate running buffer, urinary allantoin was separated from related compounds and internal standard in less than 30 min. The method was linear up to 1.25 g/L (quantification limit: 30 mg/L); precision was below 10%. The total amount of allantoin excreted in patients treated by rasburicase ranged from 1.5 g to 7.9 g/4 days. CONCLUSION: This CZE assay is a simple, rapid and reproducible method to measure allantoin in urine. Different elimination profiles have been found in patients treated with rasburicase. PMID- 16316641 TI - Abnormalities in neural crest cell migration in laminin alpha5 mutant mice. AB - Although numerous in vitro experiments suggest that extracellular matrix molecules like laminin can influence neural crest migration, little is known about their function in the embryo. Here, we show that laminin alpha5, a gene up regulated during neural crest induction, is localized in regions of newly formed cranial and trunk neural folds and adjacent neural crest migratory pathways in a manner largely conserved between chick and mouse. In laminin alpha5 mutant mice, neural crest migratory streams appear expanded in width compared to wild type. Conversely, neural folds exposed to laminin alpha5 in vitro show a reduction by half in the number of migratory neural crest cells. During gangliogenesis, laminin alpha5 mutants exhibit defects in condensing cranial sensory and trunk sympathetic ganglia. However, ganglia apparently recover at later stages. These data suggest that the laminin alpha5 subunit functions as a cue that restricts neural crest cells, focusing their migratory pathways and condensation into ganglia. Thus, it is required for proper migration and timely differentiation of some neural crest populations. PMID- 16316642 TI - Retinoic acid activates myogenesis in vivo through Fgf8 signalling. AB - Retinoic acid (RA) has been shown to regulate muscle differentiation in vitro. Here, we have investigated the role of RA signalling during embryonic myogenesis in zebrafish. We have altered RA signalling from gastrulation stages onwards by either inhibiting endogenous RA synthesis using an inhibitor of retinaldehyde dehydrogenases (DEAB) or by addition of exogenous RA. DEAB reduces expression of the myogenic markers myoD and myogenin in somites, whereas RA induces increased expression of these genes and strongly induces premature myoD expression in the presomitic mesoderm (psm). The expression dynamics of myf5 in presomitic and somitic mesoderm suggest that RA promotes muscle differentiation, a role supported by the fact that RA activates expression of fast myosin, while DEAB represses it. We identify Fgf8 as a major relay factor in RA-mediated activation of myogenesis. We show that fgf8 expression in somites and anterior psm is regulated by RA, and find that in the absence of Fgf8 signalling in the acerebellar mutant RA fails to promote myoD expression. We propose that, in the developing embryo, localised synthesis of RA by Raldh2 in the anterior psm and in somites activates fgf8 expression which in turn induces the expression of myogenic genes and fast muscle differentiation. PMID- 16316643 TI - Fog1 is required for cardiac looping in zebrafish. AB - To further our understanding of FOG gene function during cardiac development, we utilized zebrafish to examine FOG's role in the early steps of heart morphogenesis. We identified fragments of three fog genes in the zebrafish genomic database and isolated full-length coding sequences for each of these genes by using a combination of RT-PCR and 5'-RACE. One gene was similar to murine FOG-1 (fog1), while the remaining two were similar to murine FOG-2 (fog2a and fog2b). All Fog proteins were able to physically interact with GATA4 and function as transcriptional co-repressors. Whole-mount in situ hybridization revealed fog1 expression in the heart, the hematopoietic system, and the brain, while fog2a and fog2b expression was restricted to the brain. Injection of zebrafish embryos with a morpholino directed against fog1 resulted in embryos with a large pericardial effusion and an unlooped heart tube. This looping defect could be rescued by co-injection of mRNA encoding murine FOG-1, but not by mRNA encoding FOG-1 lacking the FOG repression motif. Taken together, these results demonstrate the importance of FOG proteins for zebrafish cardiac development and suggest a previously unappreciated role for FOG proteins in heart looping that is dependent on the FOG repression motif. PMID- 16316644 TI - Glucose concentration-dependent potentiation of insulin secretion by a new chemical entity, KCP256. AB - The insulinotropic activity of KCP256 [(R)-8-benzyl-2-cyclopentyl-7, 8-dihydro-4 propyl-1H-imidazo[2,1-i]purin-5(4H)-one hydrochloride] was examined using MIN6 cells (a pancreatic beta-cell line) and pancreatic islets isolated from rats. Unlike sulfonylurea anti-diabetic drugs, KCP256 dose-dependently (0.1-10 microM) enhanced insulin secretion from MIN6 cells and its insulinotropic effect was exerted only at high concentrations of glucose (8.3-22 mM) but not at low concentrations of glucose (3.3-5.5 mM). Furthermore, the action mechanism of KCP256 was different because, unlike sulfonylurea drugs, KCP256 did not displace the binding of [3H]glibenclamide, and did not inhibit the 86Rb+ efflux nor K(ATP) channel activity. In isolated islets, KCP256 also enhanced insulin secretion in a dose- and a glucose-concentration-dependent manner. Plasma levels of insulin after glucose challenge in KCP256-administrated rats were higher than those in vehicle-administrated animals, indicating that KCP256 can enhance insulin secretion in vivo. Since the insulinotropic activity of KCP256 only occurs at high concentrations of glucose, this novel drug may exhibit a decreased risk of drug-induced hypoglycemia compared with sulfonylurea drugs when treating patients with diabetes. PMID- 16316645 TI - Detailed pharmacological characterization of GT-2331 for the rat histamine H3 receptor. AB - Histamine H(3) receptor antagonists are potential therapeutic agents for cognitive dysfunction, epilepsy, hypersomnia and obesity. GT-2331 (4-[(R,R)-2 (5,5-dimethyl-1-hexynyl)cyclopropyl]-1H-imidazole) was originally identified as a potent histamine H(3) receptor antagonist. However, recent reports demonstrated a complex pharmacology for GT-2331. To further understand the pharmacological profile of GT-2331, we characterized GT-2331 using various in vitro and in vivo assays. In vitro, GT-2331 behaved as a full agonist on adenylyl cyclase inhibition and as a partial agonist on [(35)S]GTPgammaS binding at the recombinant rat histamine H(3) receptor. In contrast, in vivo, GT-2331 had no effect on brain histamine turnover while the histamine H(3) receptor agonist R alpha-methylhistamine significantly decreased histamine turnover. Furthermore, GT 2331 completely blocked R-alpha-methylhistamine-induced water intake, suggesting that GT-2331 behaves as a full antagonist. Thus, GT-2331 displayed the spectrum of pharmacological activities from full agonism to full antagonism, these observations suggest that histamine H(3) receptor ligands need to be carefully evaluated in various paradigms. PMID- 16316646 TI - Excitatory amino acid receptors are involved in morphine-induced synchronous oscillatory discharges in the locus coeruleus of rats. AB - Our previous studies demonstrated that morphine not only decreases the firing rate of locus coeruleus neurons, but that it induces synchronous oscillatory discharges in the locus coeruleus. In the present study, we examined the role of excitatory amino acid input in the mechanisms of the morphine-induced synchronous oscillation in the locus coeruleus. Using a multiple-electrode recording technique, locus coeruleus neuronal activities were recorded under halothane anesthesia in adult Sprague-Dawley rats. Among 175 locus coeruleus neurons recorded after intracerebroventricular (i.c.v.) injection of morphine (26 nmol), 88 of them exhibited both decreased firing rates and synchronous oscillatory discharges. The morphine-induced oscillation and synchrony were reversed by i.c.v. injection of the non-selective excitatory amino acid receptor antagonist kynurenic acid, the selective NMDA receptor antagonist DL-2-amino-5 phosphonopentanoic acid (AP-5), or the non-NMDA receptor antagonist 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX), but not saline vehicle. These results suggest that excitatory amino acid input contributes to the morphine-induced synchronous oscillatory activity in the locus coeruleus. The results also provide us a pharmacology tool to study the influence of blockade of the locus coeruleus synchrony on neurotransmitter release and synaptic plasticity in the locus coeruleus target areas. PMID- 16316647 TI - Characterization of muscarinic receptor binding and inhibition of salivation after oral administration of tolterodine in mice. AB - The current study was undertaken to characterize the effects of oral administration of tolterodine on muscarinic receptor binding in the bladder and submaxillary gland and on salivation in mice. In the in vitro experiment, tolterodine and its metabolite (5-hydroxymethyl metabolite: 5-HM) competed concentration-dependently with [N-methyl-(3)H]-scopolamine ([(3)H]NMS) in the mouse bladder, submaxillary gland and heart, and the potencies of both agents were greater than that of oxybutynin. After oral administration of tolterodine (6.31, 21.0 micromol/kg) and oxybutynin (76.1 micromol/kg), there was a dose and time-dependent increase in K(d) values for specific [(3)H]NMS binding in the bladder, prostate, submaxillary gland, heart, colon and lung, compared with control values, suggesting significant muscarinic receptor binding in each tissue. The K(d) increase in each tissue by oral oxybutynin reached a maximum value of 0.5 h after oral administration and then rapidly declined, while that by tolterodine was greatest 2 h after the administration and it was maintained for at least 6 or 12 h, depending on the dose and on the tissue. Thus, muscarinic receptor binding of oral tolterodine was slower in onset and of a longer duration than that of oxybutynin. Also, oral oxybutynin showed relatively greater receptor binding in the submaxillary gland as compared with other tissues, but such high selectivity to the exocrine gland muscarinic receptors was not observed by oral tolterodine. Oral administration of tolterodine and oxybutynin reduced significantly the pilocarpine-induced salivary secretion in mice, and the attenuation of oral tolterodine appeared more slowly and it was more persistent than that of oral oxybutynin. The antagonistic effect of oral tolterodine on the dose-response curves to pilocarpine was significantly weaker than that of oxybutynin. These data suggest that oral tolterodine, compared with the case of oral oxybutynin, binds more selectively to muscarinic receptors in the mouse bladder than in the submaxillary gland, which may be advantageous in treating patients with overactive bladder. PMID- 16316648 TI - A mutation in the local anaesthetic binding site abolishes toluene effects in sodium channels. AB - Toluene is a solvent of abuse that inhibits cardiac sodium channels in a manner that resembles the action of local anaesthetics. The purpose of this work was to analyze toluene effects on skeletal muscle sodium channels with and without beta1 subunit (Nav1.4+beta1 and Nav1.4-beta1, respectively) expressed in Xenopus laevis oocytes and to compare them with those produced in the F1579A mutant channel lacking a local anaesthetic binding site. Toluene inhibited Nav1.4 sodium currents (IC50=2.7 mM in Nav1.4+beta1 and 2.2 mM in Nav1.4-beta1 in a concentration dependent way. Toluene (3 mM) blocked sodium currents in Nav1.4 channels proportionally throughout the entire current-voltage relationship producing inactivation at more negative potentials. Minimal inhibition was produced by 3 mM toluene in F1579A mutant channels. Recovery from inactivation was slower both in Nav1.4 and F1579A channels in the presence of 3 mM toluene. The solvent blocked sodium currents in a use-dependent and frequency-dependent manner in Nav1.4 channels. A single mutation in the local anaesthetic binding site of Nav1.4 channels almost abolished toluene effects. These results suggest that this site is important for toluene action. PMID- 16316649 TI - GR 63799X, an EP3 receptor agonist, induced S phase arrest and 3T6 fibroblast growth inhibition. AB - The importance of arachidonic acid metabolites on the control of cell growth, particularly those derived from cyclooxygenase pathway has long been recognized. Recently, we observed that prostaglandin E(2) (PGE(2)) interaction with EP(1) and EP(4) receptors is involved in serum-induced 3T6 fibroblast growth due to their effect at various levels of the cell cycle machinery. This study shows that prostanoid EP(3) receptor was expressed in 3T6 fibroblast. We studied the role of EP(3) receptor agonist GR 63799X in serum-induced 3T6 cell proliferation. This was concentration-dependent inhibit (IC(50) approximately 10 microM) to a complete inhibition without any cytotoxic or proapoptotic effect. The prostanoid EP(3) receptor agonist treatment decreased the G(0)/G(1) and G(2)/M populations whereas cells were accumulated in S phase. This arrest in S phase was associated with a decrease in cyclin B levels and the enhancement of p21 expression. Our data show that EP(3) agonist decreases cAMP levels in our experimental conditions. Interestingly, the S arrest caused by prostanoid EP(3) receptor agonist seems to be cAMP dependent, at least in part, because forskolin treatment allowed S-arrested cells to progress through cell cycle and consequently growth. Thus, our results suggest that PGE(2) EP(3) receptor interaction may be involved in serum-induced 3T6 fibroblast growth due to their effects on cAMP levels and on cell cycle machinery of the S phase. PMID- 16316650 TI - A role for cannabinoid receptors, but not endogenous opioids, in the antinociceptive activity of the CB2-selective agonist, GW405833. AB - Several recent reports have demonstrated a role for selective cannabinoid CB2 receptor agonists in pain modulation, showing both analgesic and antihyperalgesic activities. While the mechanism of action is poorly understood, it has been postulated that these effects may be indirect, involving release of endogenous opioids. We have previously reported that administration of the selective cannabinoid CB2 receptor agonist GW405833 (2,3-dichloro-phenyl)-[5-methoxy-2 methyl-3-(2-morpholin-4-yl-ethyl)-indol-1-yl]-methanone) to rats elicits potent and efficacious antihyperalgesic effects against neuropathic and inflammatory pain and, at high dose (100 mg/kg), is analgesic and ataxic [Valenzano, K.J., Tafesse, L., Lee, G., Harrison, J.E., Boulet, J., Gottshall, S.L., Mark, L., Pearson, M.S., Miller, W., Shan, S., Rabadi, L., Rotstheyn, Y., Chaffer, S.M., Turchin, P.I., Elsemore, D.A., Toth, M., Koetzner, L., Whiteside, G.T., 2005. Pharmacological and pharmacokinetic characterization of the cannabinoid receptor 2 agonist, GW405833, utilizing rodent models of acute and chronic pain, anxiety, ataxia and catalepsy. Neuropharmacology 48, 658-672]. In the current study, we confirm these properties using mouse models and investigate the role of cannabinoid CB2 receptors using knockout animals. Furthermore, we provide evidence that the antinociceptive properties of GW405833 are opioid independent. GW405833 elicited robust antihyperalgesic effects in mouse models of inflammatory (Freund's complete adjuvant) and neuropathic (Seltzer) pain. In contrast, GW405833 showed no antihyperalgesic activity against Freund's complete adjuvant mediated inflammatory pain in cannabinoid CB2 receptor knockout mice. As in rats, high-dose GW405833 (100 mg/kg) showed both analgesic and sedative activities in wild-type mice, activities that were also apparent in cannabinoid CB2 receptor knockout mice. In rats, neither the antihyperalgesic effect in the Freund's complete adjuvant model nor the analgesic effects in tail flick and hot plate assays were inhibited by pre-treatment with the non-selective opioid receptor antagonist, naltrexone. These data demonstrate that the antihyperalgesic effects of GW405833 are mediated via the cannabinoid CB2 receptor, whereas the analgesic and sedative effects are not. Furthermore, these data suggest that the mechanism of action for GW405833 does not depend on the release of endogenous opioids. PMID- 16316651 TI - Pharmacological characterization of the active synthetic factor Xa inhibitors M55551 and M55165. AB - Factor Xa plays an important role in blood coagulation and is widely regarded as an attractive target for antithrombotic drug development. M55551 and M55165 (1 arylsulfonyl-3-piperazinone derivatives) are novel synthetic factor Xa inhibitors. In vitro, M55551 and M55165 competitively inhibited factor Xa with K(i) values of 3.2 nM and 2.3 nM, respectively, and prolonged clotting time in human and rat plasma. Pharmacokinetic analysis of these compounds revealed that M55551 was intravenously active with a short half-life (0.2 h) and that M55165 exhibited good bioavailability (31%) with a long half-life (3.9 h). Therefore, the antithrombotic effects of M55551 and M55165 were compared with those of the intravenous anticoagulant argatroban and the oral anticoagulant warfarin. Intravenous administration of M55551 and oral administration of M55165 inhibited thrombus formation at 0.3 mg/kg and 10 mg/kg, respectively, without significant prolongation of bleeding time. In contrast, although argatroban (0.3 mg/kg) and warfarin (1 mg/kg) also inhibited thrombus formation, significant prolongation of bleeding time was observed at dosages of 3 mg/kg and 1 mg/kg, respectively. These results suggest that M55551 and M55165 are potent factor Xa inhibitors that are active upon intravenous and oral administration, respectively, and that may prove clinically useful for the treatment of thrombosis while minimizing bleeding risks. PMID- 16316652 TI - Quercetin, a bioflavonoid, accelerates TNF-alpha-induced growth inhibition and apoptosis in MC3T3-E1 osteoblastic cells. AB - The bioflavonoid quercetin is believed to play an important role in preventing bone loss by affecting osteoclastogenesis and regulating many systemic and local factors including hormones and cytokines. This study examined how quercetin acts on tumor necrosis factor-alpha (TNF-alpha)-mediated growth inhibition and apoptosis in MC3T3-E1 osteoblastic cells. Tritium uptake assay showed that a quercetin treatment accelerated TNF-alpha-induced inhibition of DNA synthesis in the cells in a dose-dependent manner. Both the 3-(4,5-dimethylthiazol-2yl-)-2,5 diphenyl tetrazolium bromide and trypan blue staining assays also showed the quercetin-mediated facilitation of TNF-alpha-induced cytotoxicity in the cells. Apoptosis assays revealed an accelerating effect of quercetin on TNF-alpha induced apoptosis in MC3T3-E1 cells. In addition, Fas activation and poly (ADP ribose) polymerase cleavage are thought to be closely associated with the TNF alpha-induced apoptosis and its acceleration by the quercetin treatment in the cells. Collectively, this study showed that quercetin accelerates the TNF-alpha induced growth inhibition and apoptosis in MC3T3-E1 osteoblastic cells. PMID- 16316653 TI - Cannabinoid CB2 receptor agonist activity in the hindpaw incision model of postoperative pain. AB - The identification of peripherally expressed CB2 receptors and reports that the selective activation of cannabinoid CB2 receptors produces antinociception without traditional cannabinergic side effects suggests that selective cannabinoid CB2 receptor agonists might be useful in the management of pain. In a rat hindpaw incision model, we examined the antiallodynic activity of the selective cannabinoid CB2 receptor agonists AM1241 (3-30 mg/kg i.p.), GW405833 (3 30 mg/kg i.p.), and HU-308 (0.3-30 mg/kg i.p.). The rank order for efficacy in the hindpaw incision model following a dose of 10 mg/kg, i.p. was AM1241 > GW405833 = HU-308, and the selective cannabinoid CB2 receptor antagonist, SR144528, reversed the antiallodynic effect of HU-308. Together, these data suggest that selective cannabinoid CB2 receptor agonists might represent a new class of postoperative analgesics. PMID- 16316654 TI - Pharmacogenomics and cardiovascular drugs: need for integrated biological system with phenotypes and proteomic markers. AB - Personalized medicine is based on a better knowledge of biological variability, considering the important part due to genetics. When trying to identify involved genes and their products in differential cardiovascular drug responses, a five step strategy is to be followed: 1) Pharmacokinetic-related genes and phenotypes (2) Pharmacodynamic targets, genes and products (3) Cardiovascular diseases and risks depending on specific or large metabolic cycles (4) Physiological variations of previously identified genes and proteins (5) Environment influences on them. After summarizing the most well-known genes involved in drug metabolism, we will take as example of drugs, the statins, considered as very important drugs from a Public-Health standpoint, but also for economical reasons. These drugs respond differently in human depending on multiple polymorphisms. We will give examples with common ApoE polymorphisms influencing the hypolipemic effects of statins. These drugs also have pleiotropic effects and decrease inflammatory markers. This illustrates the need to separate clinical diseases phenotypes in specific metabolic pathways, which could propose other classifications, of diseases and related genes. Hypertension is also a good example of clinical phenotype which should be followed after various therapeutic approaches by genes polymorphisms and proteins markers. Gene products are under clear environmental expression variations such as age, body mass index and obesity, alcohol, tobacco and dietary interventions which are the first therapeutical actions taken in cardiovascular diseases. But at each of the five steps, within a pharmacoproteomic strategy, we also need to use available information from peptides, proteins and metabolites, which usually are the gene products. A profiling approach, i.e., dealing with genomics, but now also with proteomics, is to be used. In conclusion, the profiling, as well as the large amount of data, will more than before render necessary an organized interpretation of DNA, RNA as well as proteins variations, both at individual and population level. PMID- 16316655 TI - Development of antinociceptive tolerance and physical dependence following morphine i.c.v. infusion in mice. AB - The chronic i.c.v. infusion of morphine has been reported for rats but not for mice. In the current report, the antinociceptive tolerance to both i.c.v. morphine infusion and s.c. implantation of morphine pellets in mice was compared. Physical dependence after i.c.v. morphine infusion was also evaluated. Osmotic minipumps were filled with morphine (50 mM), connected to i.c.v. cannulae, and implanted s.c. to deliver 50 nmol/h for 3 days (i.e., 3.6 micromol total). Robust jumping precipitated by naloxone (1 mg/kg, s.c.) indicated the development of physical dependence. Tolerance to i.c.v., i.t., and i.v. morphine (6.3-, 2.0-, and 4.4-fold, respectively) was observed using the tail flick test. Mice implanted with pellets containing 75 mg morphine for 3 days (i.e., approximately 260 micromol total) were also tolerant to morphine (6.5-, 7.5- and 18-fold, respectively). Thus, the tolerance developed using the two methods was not identical. These results allow comparison of morphine tested by 3 different routes (i.c.v., i.t., and i.v.) after chronic morphine treatment by two routes (i.c.v. and s.c.) in a single study. PMID- 16316656 TI - Lack of direct role of parkin in the steady-state level and aggregation of alpha synuclein and the clearance of pre-formed aggregates. AB - Mutations in parkin and alpha-synuclein (alpha-syn) are linked to heritable forms of Parkinson's disease (PD). Recently, it has been shown that parkin mitigates alpha-syn-induced neuronal cell death in animal and tissue culture models, suggesting that there is a functional relationship between these two proteins. Although the mechanism by which parkin protects cells from alpha-syn-induced cytotoxicity remains elusive, it is tempting to speculate that parkin might directly regulate the normal metabolism and aggregation of alpha-syn. In the current study, we show that neither the suppression of endogenous parkin expression nor ectopic overexpression affects the steady-state levels of endogenous alpha-syn expression, overall aggregation of this protein, or breakdown of pre-formed aggregates in human neuroblastoma cells. These results suggest that parkin is not directly involved in the metabolism of alpha-syn, its aggregation, or the clearance of pre-formed aggregates. PMID- 16316657 TI - Correlation of 8- and 12-h urinary protein with 24-h urinary protein in pre eclampsia. PMID- 16316658 TI - Associations between physical activity, peripheral atherosclerosis and bone status in healthy Japanese women. AB - The aim of this cross-sectional study was to investigate whether physical activity and bone status may affect arterial thickening and stiffening in healthy Japanese women. Healthy women (n = 149; mean age, 54 years) were recruited from those who participated in a local health check program at the Osaka City University Hospital. Physical activity was assessed by physical functioning score of SF-36, and bone status by bone mineral density (BMD) in lumbar spine and calcaneus osteo-sono index (OSI). Arterial wall thickening assessed by intima media thickness (IMT) in common carotid artery (CA) and femoral artery (FA), and arterial wall stiffening by peak wave velocity (PWV) in heart-carotid (hc) and heart-femoral (hf) as central segment and in heart-brachial (hb) and femoral ankle (fa) as peripheral segment, respectively. By Spearman Rank correlation, lumbar spine BMD was correlated negatively with CA IMT (rho = -0.225, p < 0.05) and FA IMT (rho = -0.215, p < 0.05), and calcaneus OSI with FA IMT (rho = -0.330, p < 0.0001) but not CA IMT (rho = -0051, p = 0.5335). Both lumbar spine BMD and calcaneus OSI correlated negatively with PWV in all segments (all p < 0.05). Physical functioning score correlated weakly but significantly in a negative manner with all PWV segments (all p < 0.05) but not IMT. Multiple regression analyses revealed a significant association of calcaneus OSI (beta = -0.240, p = 0.0039) but not lumbar spine BMD (beta = -0.067, p = 0.4541) with FA IMT, although neither lumbar spine BMD nor calcaneus OSI was associated with CA IMT. Furthermore, physical functioning score was independently associated with hb and fa PWV but not hc and hf PWV, suggesting the preferential association with peripheral segment including lower extremities. Neither lumbar spine BMD nor calcaneus OSI was associated with any segment of PWV. In conclusion, it was suggested that calcaneus OSI might be associated with arterial wall thickening preferentially in femoral artery, and that physical activity may be associated with arterial wall stiffening in peripheral segment including lower extremity but not in central segment in healthy Japanese women. PMID- 16316659 TI - A nonlinear biphasic viscohyperelastic model for articular cartilage. AB - Experiments on articular cartilage have shown nonlinear stress-strain curves under finite deformations as well as intrinsic viscous effects of the solid phase. The aim of this study was to propose a nonlinear biphasic viscohyperelastic model that combines the intrinsic viscous effects of the proteoglycan matrix with a nonlinear hyperelastic constitutive equation. The proposed equation satisfies objectivity and reduces for uniaxial loading to a solid type viscous model in which the actions of the springs are represented by the hyperelastic function proposed by Holmes and Mow [1990. J. Biomechanics 23, 1145-1156.]. Results of the model, that were efficiently implemented in an updated Lagrangian algorithm, were compared with experimental infinitesimal data reported by DiSilverstro and Suh [2001. J. Biomechanics 34, 519-525.] and showed acceptable fitting for the axial force (R(2)=0.991) and lateral displacement (R(2)=0.914) curves in unconfined compression as well as a good fitting of the axial indentation force curve (R(2)=0.982). In addition, the model showed an excellent fitting of finite-deformation confined compression stress relaxation data reported by Ateshian et al. [1997. J. Biomechanics 30, 1157-1164.] and Huang et al. [2005. J. Biomechanics 38, 799-809.] (R(2)=0.993 and R(2)=0.995, respectively). The constitutive equation may be used to represent the mechanical behavior of the proteoglycan matrix in a fiber reinforced model of articular cartilage. PMID- 16316660 TI - iPLA2 inhibitor blocks negative inotropic effect of HIV gp120 on cardiac myocytes. AB - Recent improvements in survival from AIDS have been accompanied by an increased recognition of the potential importance of other manifestations of HIV infection, including cardiomyopathy. Mechanisms responsible for HIV cardiomyopathy are unknown, but may include direct effects of HIV proteins on the heart. We previously provided support for direct effects of HIV proteins by demonstrating a negative inotropic effect of the HIV coat protein, gp120, on isolated adult rat ventricular myocytes (ARVM). We now report that this negative inotropic effect of HIV gp120 is mediated by a signaling pathway involving p38 MAP kinase, iPLA2 and troponin I. Exposure of ARVM to HIV gp120 resulted in maximal activation of iPLA2 by 60 min as reflected in hydrolysis of arachidonyl thiophosphatidylcholine that was completely blocked by the iPLA2 inhibitor, bromoenol lactone (BEL) or the p38 MAP kinase inhibitor, SB203580. The negative inotropic effect of gp120 was blocked by BEL, as well as SB203580. BEL did not block gp120 stimulated phosphorylation of p38 MAP kinase itself, and/or its downstream effectors, ATF2 or MAPKAP2. However, BEL did block gp120-stimulated phosphorylation of troponin I. Thus, the negative inotropic effect of HIV gp120 requires activation of p38 MAP kinase and iPLA2; as well as troponin I phosphorylation. Activation of this novel p38 MAP kinase-iPLA2-troponin I signaling pathway may contribute to HIV cardiomyopathy. PMID- 16316661 TI - Crystal structure of penicillin-binding protein 1a (PBP1a) reveals a mutational hotspot implicated in beta-lactam resistance in Streptococcus pneumoniae. AB - Streptococcus pneumoniae is a major human pathogen whose infections have been treated with beta-lactam antibiotics for over 60 years, but the proliferation of strains that are highly resistant to such drugs is a problem of worldwide concern. Beta-lactams target penicillin-binding proteins (PBPs), membrane associated enzymes that play essential roles in the peptidoglycan biosynthetic process. Bifunctional PBPs catalyze both the polymerization of glycan chains (glycosyltransfer) and the cross-linking of adjacent pentapeptides (transpeptidation), while monofunctional enzymes catalyze only the latter reaction. Although S. pneumoniae has six PBPs, only three (PBP1a, PBP2x, PBP2b) are major resistance determinants, with PBP1a being the only bifunctional enzyme. PBP1a plays a key role in septum formation during the cell division cycle and its modification is essential for the development of high-level resistance to penicillins and cephalosporins. The crystal structure of a soluble form of pneumococcal PBP1a (PBP1a*) has been solved to 2.6A and reveals that it folds into three domains. The N terminus contains a peptide from the glycosyltransfer domain bound to an interdomain linker region, followed by a central, transpeptidase domain, and a small C-terminal unit. An analysis of PBP1a sequences from drug-resistant clinical strains in light of the structure reveals the existence of a mutational hotspot at the entrance of the catalytic cleft that leads to the modification of the polarity and accessibility of the mutated PBP1a active site. The presence of this hotspot in all variants sequenced to date is of key relevance for the development of novel antibiotherapies for the treatment of beta-lactam-resistant pneumococcal strains. PMID- 16316662 TI - IL-15 is elevated in serum and cerebrospinal fluid of patients with multiple sclerosis. AB - Interleukin-15 (IL-15) is a novel proinflammatory cytokine having similar biological activities to IL-2 which is implicated in the pathogenesis of multiple sclerosis. It is produced by activated blood monocytes, macrophages and glial cells. There is little information about the involvement of IL-15 in the development of multiple sclerosis (MS). The objective of our study was to measure IL-15 serum and cerebrospinal fluid (CSF) levels in MS patients and to correlate serum and CSF IL-15 concentrations with clinical parameters of the disease. CSF IL-15/Serum IL-15 ratio (c/s IL-15 ratio) was introduced to assess the origin of elevated IL-15 levels. MATERIALS AND METHODS: We measured serum and CSF IL-15 levels in 52 patients with MS and 36 age and gender matched patients with inflammatory (IND) and non-inflammatory neurological diseases (NIND) studied as control groups. IL-15 levels were correlated with clinical parameters as duration, disability, MRI activity and clinical subtypes of the disease. RESULTS: MS patients were found to have significantly higher serum IL-15 levels compared with IND (p=0.00016) and NIND patients (p=0.00045). Elevated levels of IL-15 were also found in CSF samples from MS patients compared with patients with IND (p=0.00034) and NIND (p=0.0003). Among MS subgroups there were no statistically different IL-15 serum and CSF concentrations. No significant correlation of serum and CSF IL-15 concentrations with MRI activity, disability assessed by EDSS score and duration of the disease were also found. C/S IL-15 ratio was found lower in MS patients compared with IND (p=0.01) and not significantly different compared with NIND patients (p=0.14) suggesting that systemic activation might be the source of high CSF IL-15 levels in MS patients. CONCLUSIONS: Our findings suggest a possible role of IL-15 in the immunopathogenetic mechanisms of MS. PMID- 16316663 TI - Could neonatal testosterone replacement prevent alterations induced by prenatal stress in male rats? AB - The present study was designed to examine whether testosterone replacement is able to prevent some effects of maternal restraint stress--during the period of brain sexual differentiation--on endocrine system and sexual behavior in male rat descendants. Pregnant rats were exposed to restraint stress for 1 h/day from gestational days 18 to 22. At birth, some male pups from these stressed rats received testosterone propionate. The neonatal testosterone replacement was able to prevent the reduction in anogenital distance at 22 days of age observed in pups from stressed pregnant rats as well as prevents the decrease in testosterone levels during the adulthood of these animals. Testosterone replacement in these males also presented an improvement in sexual performance. In this way, testosterone replacement probably through increasing neonatal level of this hormone was able to prevent the later alterations caused by the prenatal stress during the period of brain sexual differentiation. PMID- 16316664 TI - The preventive effect of vitamin D3 on radiation-induced hair toxicity in a rat model. AB - Our aim is to investigate the protective effect of vitamin D3 especially from radiation-induced hair toxicity. A model of skin radiation injury was developed and a single fraction of 20 Gy Gamma irradiation was applied to the right dorsal skin of fourteen rats. All animals were randomly divided into 2 groups: Group I: irradiation alone (n = 7) and Group II: irradiation and 0.2 microg vitamin D3 given IM (n = 7). Fifty days after post-irradiation rats were sacrificed. The outcomes were evaluated on the basis of histopathological findings and immunohistochemical staining for Vitamin D receptor (VDR) in skin and hair follicles. The number of hair follicles in the radiation field for the group of animals irradiated without pretreatment was significantly lower than outside of the irradiated area (p = 0.016) as it is expected. Contrarily the number of hair follicles did not show significant difference in the pretreated group between the irradiated field and outside of the fields (p = 0,14). Skin of the vitamin D3 pretreated group demonstrated stronger immunoreactivity for VDR compared to irradiation alone group. These results indicate that administration of vitamin D3 may protect hair follicles from radiation toxicity. Further clinical trials should be conducted to prove the preventive effect of vitamin D3 as well as dosing and timing of the agent on radiation-induced alopecia. PMID- 16316665 TI - Induction of apoptosis by pterocarpans from Platymiscium floribundum in HL-60 human leukemia cells. AB - (+)-2,3,9-Trimethoxy-pterocarpan (1) (+)-3,9-dimethoxy-pterocarpan [(+) homopterocarpin] (2), (+)-3-hydroxy-9-methoxy-pterocarpan [(+)-medicarpin] (3) and (+)-3,4-dihydroxy-9-methoxy-pterocarpan [(+)-vesticarpan] (4) are cytotoxic pterocarpans isolated from the native Brazilian plant Platymiscium floribundum. The purpose of the present study was to examine whether induction of apoptosis and/or inhibition of DNA synthesis is involved in the cytotoxicity of these pterocarpans in human leukemia cells. The effect on cell viability determined using the trypan exclusion assay revealed that all compounds tested reduced the number of viable cells, while only in the presence of 3 and 4, there was an increase of nonviable cells. The analysis of membrane integrity and morphological modifications by flow cytometry in the presence of these two compounds indicated that treated cells undergo necrosis, while 1 and 2 trigger apoptosis. DNA synthesis seemed to be affected since BrdU incorporation was inhibited in a dose dependent manner in the presence of all tested compounds. Pterocarpan treatment also induced an increase in the amount of subdiploid DNA, indicating internucleosomal DNA breakdown, mitochondrial depolarization and caspase-3 activation, which indicate apoptosis induction. PMID- 16316666 TI - Polychlorinated biphenyls (PCBs) in marine organisms from four harbours in Guam. PMID- 16316667 TI - A call for standardized protocol of coral recruitment research and outlines for its conception. PMID- 16316668 TI - Quantifying the L-arginine paradox in vivo. AB - NO and PO(2) microelectrodes were used to quantify the effects of increased availability of L-arginine in an exteriorized rat mesentery and small intestine microcirculatory preparation in n = 16 rats. During short periods of elevated L arginine added to the superfusion bath, transient changes in perivascular NO or PO(2) were measured at 171 perivascular sites near intestinal arterioles and venules, simultaneously with tissue perfusion using laser Doppler flowmetry (LDF). Excess L-arginine increased perivascular NO over twofold, by 411 +/- 42 nM above the baseline of 329 +/- 30 nM (P < 0.0001), and increased tissue perfusion by 35.5 +/- 7.5% (P < 0.0001). No difference between arterioles and venules was observed in the magnitude or time course of the NO responses. Both increases and decreases in perivascular PO(2) were observed after excess L-arginine, with a similar increase in tissue perfusion by 42.0 +/- 12.3% (P < 0.0001). Our NO measurements confirm that increased bioavailability of L-arginine causes a significant increase in NO production throughout the microcirculation of this preparation, with increased tissue perfusion, and provides direct in vivo evidence for the L-arginine paradox. PMID- 16316669 TI - Endocannabinoids at the spinal level regulate, but do not mediate, nonopioid stress-induced analgesia. AB - Recent work in our laboratories has demonstrated that an opioid-independent form of stress-induced analgesia (SIA) is mediated by endogenous cannabinoids [Hohmann et al., 2005. Nature 435, 1108]. Non-opioid SIA, induced by a 3-min continuous foot shock, is characterized by the mobilization of two endocannabinoid lipids--2 arachidonoylglycerol (2-AG) and anandamide--in the midbrain periaqueductal gray (PAG). The present studies were conducted to examine the contributions of spinal endocannabinoids to nonopioid SIA. Time-dependent increases in levels of 2-AG, but not anandamide, were observed in lumbar spinal cord extracts derived from shocked relative to non-shocked rats. Notably, 2-AG accumulation was of smaller magnitude than that observed previously in the dorsal midbrain following foot shock. 2-AG is preferentially degraded by monoacylglycerol lipase (MGL), whereas anandamide is hydrolyzed primarily by fatty-acid amide hydrolase (FAAH). This metabolic segregation enabled us to manipulate endocannabinoid tone at the spinal level to further evaluate the roles of 2-AG and anandamide in nonopioid SIA. Intrathecal administration of the competitive CB1 antagonist SR141716A (rimonabant) failed to suppress nonopioid SIA, suggesting that supraspinal rather than spinal CB1 receptor activation plays a pivotal role in endocannabinoid mediated SIA. By contrast, spinal inhibition of MGL using URB602, which selectively inhibits 2-AG hydrolysis in the PAG, enhanced SIA through a CB1 selective mechanism. Spinal inhibition of FAAH, with either URB597 or arachidonoyl serotonin (AA-5-HT), also enhanced SIA through a CB1-mediated mechanism, presumably by increasing accumulation of tonically released anandamide. Our results suggest that endocannabinoids in the spinal cord regulate, but do not mediate, nonopioid SIA. PMID- 16316670 TI - Use of an inverse agonist radioligand [3H]A-317920 reveals distinct pharmacological profiles of the rat histamine H3 receptor. AB - Selective radioligands for histamine H(3) receptors have been used to characterize H(3) receptor pharmacology by radioligand binding assays and to determine H(3) receptor distribution by tissue autoradiography. Here we report the synthesis and receptor binding characterization of [(3)H]A-317920 (furan-2 carboxylic acid(2-[4-[3-([3,5-(3)H]4-cyclopropanecarbonyl-phenoxy)-propyl] piperazin-1-yl]-1-methyl-2-oxo-ethyl)-amide), a high affinity inverse agonist radioligand for the rat H(3) receptor. The binding of [(3)H]A-317920 to rat cortical and cloned H(3) receptors revealed fast on- and slower off-rate kinetics with calculated K(d) values in agreement with those determined in saturation binding assays (0.2 nM for both receptors). Further, we compared [(3)H]A-317920 with the agonist [(3)H](N)-alpha-methylhistamine ([(3)H]NalphaMH) as radioligand tools to study receptor pharmacology. Agonists and antagonists displaced [(3)H]NalphaMH with one-site binding characteristics and Hill slopes approached unity. In contrast, although antagonists exhibited one-site binding, [(3)H]A 317920 displacement by agonists was best fit by two-site binding models, and the potencies of the high affinity, GDP-sensitive sites correlated with the potencies defined in [(3)H]NalphaMH binding. Unlike [(125)I]iodoproxyfan, [(3)H]A-317920 exhibits potent and selective binding to rat H(3) receptors with low binding to non-H(3) sites, including cytochrome P450. These findings show that [(3)H]A 317920 is a potent rat H(3) receptor antagonist radioligand and has utility for studying H(3) receptor pharmacology. PMID- 16316671 TI - An epidemiological study of familial neurocysticercosis in an endemic Mexican community. AB - Neurocysticercosis (NC) caused by Taenia solium is a frequent parasitic disease of the central nervous system. It is highly endemic in many developing countries, where many people are exposed but few become infected. Here, the relevance of age, gender, and genetic and exposure factors on NC susceptibility was studied in 649 inhabitants of a rural community of Mexico. Endemicity was confirmed by the high prevalence of pig cysticercosis (32.8%) and human seroprevalence (43.8%). Human NC cases were diagnosed by computerised tomography scans. A questionnaire to evaluate risk factors was applied and familial relationships between participants were registered. An overall NC frequency of 9.1% (59/649) was found. NC frequency increased with age but did not associate with gender. Most NC cases were asymptomatic. None of the evaluated risk factors were associated with NC. No familial aggregation was detected when studying all cases, although a significant relationship between mother and child in cases with multiple parasites was found. These findings point to the fact that human NC in high exposure conditions is not simply related to exposure factors and they do not support the participation of a major gene in single-cyst NC. Rather, our results point to a complex interaction of genetic and environmental factors involved in NC. PMID- 16316672 TI - In vitro binding of anthrax protective antigen on bacteriophage T4 capsid surface through Hoc-capsid interactions: a strategy for efficient display of large full length proteins. AB - An in vitro binding system is described to display large full-length proteins on bacteriophage T4 capsid surface at high density. The phage T4 icosahedral capsid features 155 copies of a nonessential highly antigenic outer capsid protein, Hoc, at the center of each major capsid protein hexon. Gene fusions were engineered to express the 83-kDa protective antigen (PA) from Bacillus anthracis fused to the N terminus of Hoc and the 130-kDa PA-Hoc protein was expressed in Escherichia coli and purified. The purified PA-Hoc was assembled in vitro on hoc(-) phage particles. Binding was specific, stable, and of high affinity. This defined in vitro system allowed manipulation of the copy number of displayed PA and imposed no significant limitation on the size of the displayed antigen. In contrast to in vivo display systems, the in vitro approach allows all the capsid binding sites to be occupied by the 130-kDa PA-Hoc fusion protein. The PA-T4 particles were immunogenic in mice in the absence of an adjuvant, eliciting strong PA-specific antibodies and anthrax lethal toxin neutralizing antibodies. The in vitro display on phage T4 offers a novel platform for potential construction of customized vaccines against anthrax and other infectious diseases. PMID- 16316673 TI - Cell-to-cell movement of Alfalfa mosaic virus can be mediated by the movement proteins of Ilar-, bromo-, cucumo-, tobamo- and comoviruses and does not require virion formation. AB - RNA 3 of Alfalfa mosaic virus (AMV) encodes the movement protein (MP) and coat protein (CP). Chimeric RNA 3 with the AMV MP gene replaced by the corresponding MP gene of Prunus necrotic ringspot virus, Brome mosaic virus, Cucumber mosaic virus or Cowpea mosaic virus efficiently moved from cell-to-cell only when the expressed MP was extended at its C-terminus with the C-terminal 44 amino acids of AMV MP. MP of Tobacco mosaic virus supported the movement of the chimeric RNA 3 whether or not the MP was extended with the C-terminal AMV MP sequence. The replacement of the CP gene in RNA 3 by a mutant gene encoding a CP defective in virion formation did not affect cell-to-cell transport of the chimera's with a functional MP. A GST pull-down technique was used to demonstrate for the first time that the C-terminal 44 amino acids of the MP of a virus belonging to the family Bromoviridae interact specifically with AMV virus particles. Together, these results demonstrate that AMV RNA 3 can be transported from cell-to-cell by both tubule-forming and non-tubule-forming MPs if a specific MP-CP interaction occurs. PMID- 16316674 TI - Bioremediation of crystal violet using air bubble bioreactor packed with Pseudomonas aeruginosa. AB - Seven water and sediment samples were collected and tested for decolorizing crystal violet. Pseudomonas aeruginosa was the most effective isolate for dye decolorization. The LC50 of the crystal violet (115 mg/l) was measured using Artemia salina as a biomarker. The effect of different heavy metals on crystal violet decolorization was investigated. Cd2+ and Fe3+ ions showed marginal enhancement of the decolorization process, the rate was 1.35 mg/l/h compared to (1.25 mg/l/h) for the control. Phenol and m-cresol showed no effect on crystal violet decolorization, meanwhile p-cresol and p-nitrophenol reduced the decolorization rate to 1.07 and 0.01 mg/l/h, respectively. P. aeruginosa cells were immobilized by entrapment in agar-alginate beads. The beads were cultivated and reused in Erlenmeyer flask and in an air bubble column bioreactor and they enhanced the crystal violet decolorization rate to 3.33 and 7.5 mg/l/h, respectively. PMID- 16316675 TI - Development of an advanced biological treatment system applied to the removal of nitrogen and phosphorus using the sludge ceramics. AB - To develop a method of forming lake sediment into sludge ceramics with porosity and good biological adhesion for use as a medium for microorganisms in wastewater treatment, a study of the effects of forming conditions was conducted by adjusting the water content of sludge and compounding some additives. By adjusting the water content of the raw material at the kneading/pelletizing step to 40-42% and adding 3% waste glass to the raw materials to make up for the lack of flux, a sludge ceramic with a density in terms of specific gravity of saturated surface dry aggregate of about 1400 kg m(-3) was formed. In addition, to develop a small-scale wastewater treatment system capable of removing nitrogen and phosphorus, a sludge ceramic was applied as a medium for biological filtration. The results indicated that the BOD removal nitrification rate were superior to those of conventional ceramic media, reached at 95.3% and 87.4%, respectively. The introduction of iron electrolysis resulted in high treatment performance achieving BOD levels of 10 mg L(-1) or less, T-N of 10 mg L(-1) or less and T-P of 1 mg L(-1) or less. PMID- 16316676 TI - Creating and using the urgent metadata catalogue and thesaurus. AB - The Urban Regeneration and the Environment Research Programme (URGENT) required a system for cataloguing its datasets and enabling its scientific community to discover what data were available to it. This community was multidisciplinary in nature and therefore needed a range of facilities for searching. Of particular importance were facilities to help those unfamiliar with specialist terminology. To meet these needs, four applications were designed and developed: a Metadata Capture Tool for describing datasets in compliance with the National Geospatial Data Framework (NGDF) standard, a Term Entry Tool for creating an ISO compliant thesaurus, a Thesaurus Builder for merging thesauri and a Search Tool. To encourage users to help in cataloguing data, the capture tools were written as stand alone applications, which users could keep and use to build their own metadatabases. The tools contained export and import facilities that allowed the URGENT Data Centre to build a central database and publish it upon the web. During the development work, it was found necessary to extend the NGDF standard as it could not adequately describe time variant or 3-D atmospheric datasets. The four applications met their design objectives. However, a number of ergonomic issues will need to be addressed if the system is to meet the needs of the much larger up coming programmes. The main challenges will be moving from the NGDF standard to the ISO standard, hence bringing the work into line with the recommendations of the INSPIRE Project, and merging the metadatabase with the scientific database, which enable metadata maintenance to be semi-automated. PMID- 16316677 TI - Metabolites from the biodegradation of di-ester plasticizers by Rhodococcus rhodochrous. AB - The plasticizers di-2-ethylhexyl phthalate (DEHP) and di-2-ethylhexyl adipate (DEHA) are ubiquitous in the environment and undergo partial biodegradation in the presence of soil micro-organisms. The validity of a proposed pathway for the degradation of these plasticizers by Rhodococcus rhodochrous has been confirmed by the identification of 2-ethylhexanal in gas phase emissions. Complete analyses of the aqueous and gas phases were able to account for more than 98% of the 2 ethylhexanol component of the DEHA added at the beginning of a growth study. Of this, 25% was either 2-ethylhexanol or 2-ethylhexanal that had been stripped into the gas phase and, at most, only 2% of the 2-ethylhexanol component could have been removed by mineralization. It is concluded that plasticizers are of significant environmental concern due to the resistance of the metabolites to biodegradation and their known health impacts. Two of the metabolites are of added concern due to their volatility and their potential impact on indoor air quality. PMID- 16316678 TI - Leaching of heavy metals (Cu, Ni and Zn) and organic matter after sewage sludge application to Mediterranean forest soils. AB - In Mediterranean dry and semiarid areas, soil organic matter is often depleted due to ancient and intensive human activity. Under these conditions the use of sewage sludge as a land reclamation technique may be a means to revert desertification processes and to enhance soil function and nutrient cycling. However, applications of heavy metal-contaminated sewage sludges can significantly increase potentially toxic metal concentrations in soils and metal transfer to freshwater and plants. The aims of this study are 1) to investigate the leaching of Cu, Zn and Ni from three contrasted Mediterranean forest soils (a basic loam, a basic clay, and an acid loam) treated with sewage sludges and 2) to explore the relationships between metal mobility and soil properties and with the leaching of organic matter. The selected soils were incubated in columns (5 replicatesx3 soilsx3 treatments). Treatments were (a) soil application of low metal content sewage sludge (LMS), (b) soil application of metal-enriched sewage sludge (MES), and (c) control. The sewage sludge application represented a dose of 6 kg dry weight m-2. Soil columns were incubated at room temperature for 110 days and were irrigated weekly with deionised water to make a total of a 1130 mm. Leachates were collected and analysed for pH, EC, organic carbon Cu, Ni and Zn concentrations. The concentration of metals and organic matter in the leachates depended on the soil characteristics and on the type of sewage sludge added to the soil. Basic soils with a high amount of clay showed the highest metal retention capacity, while acid soils with low clay content showed the lowest. Of the three metals studied, Ni exhibited the greatest mobility. Zn mobility was also rather high, particularly in the acid soil. Despite the fact that basic soils showed greater OM content than the acid soil, organic carbon in leachates after sludge addition was of the same order of magnitude in all studied soils. OM mobility may enhance the leaching of metals while the OM bound to soil particles may enhance the retention of metals. The rate of leaching per unit of metal in the soil, for Cu in all soils and for Zn in the basic soils, did not increase even in soils amended with metal-enriched sewage sludge. However, the total amount of Cu, Zn, and Ni leached through the 30 cm columns of the metal-enriched sewage sludge increased in comparison to the control soils. In spite of this fact, metal concentrations were in most cases well below limit for drinking water quality criteria of Spanish legislation. Thus, after one single addition of metal enriched sewage sludge, for the studied Mediterranean soils, the risk of heavy metal pollution of groundwater appears to be low. PMID- 16316679 TI - A structural interpretation of voltage-gated potassium channel inactivation. AB - After channel activation, and in some cases with sub-threshold depolarizing stimuli, Kv channels undergo a time-dependent loss of conductivity by a family of mechanisms termed inactivation. To date, all identified inactivation mechanisms underlying loss of conduction in Kv channels appear to be distinct from deactivation, i.e. closure of the voltage-operated activation gate by changes in transmembrane voltage. Instead, Kv channel inactivation entails entry of channels into a stable, non-conducting state, and thereby functionally reduces the availability of channels for opening. That is, if a channel has inactivated, some time must expire after repolarization of the membrane voltage to allow the channel to recover and become available to open again. Dramatic differences between Kv channel types in the time course of inactivation and recovery underlie various roles in regulating cellular excitability and repolarization of action potentials. Therefore, the range of inactivation mechanisms exhibited by different Kv channels provides important physiological means by which the duration of action potentials in many excitable tissues can be regulated at different frequencies and potentials. In this review, we provide a detailed discussion of recent work characterizing structural and functional aspects of Kv channel gating, and attempt to reconcile these recent results with classical experimental work carried out throughout the 1990s that identified and characterized the basic mechanisms and properties of Kv channel inactivation. We identify and discuss numerous gaps in our understanding of inactivation, and review them in the light of new structural insights into channel gating. PMID- 16316680 TI - Leukaemia inhibitory factor enhances sheep fertilization in vitro via an influence on the oocyte. AB - LIF is twice transiently expressed in the mouse uterus, first at the time of ovulation and again just prior to implantation, and studies have demonstrated a beneficial influence of this cytokine on embryo development in several species. We have investigated the effect of LIF on gametes in vitro, on the hypothesis that the ovulatory peak of LIF can exert an influence on gametes present within the oviduct. We also investigated the effect of LIF on in vitro fertilization and embryo development, in oocytes from adult sheep and from prepubertal lambs that lack the preovulatory hormone surge and that are unable to sustain early embryonic development. A higher rate of pronuclear-stage embryos derived from both, adult and prepubertal female, was obtained when in vitro fertilization was performed in the presence of LIF, and there was an improved cleavage of parthenogenetic embryos when incubated with LIF immediately following activation. In contrast, LIF was found to have no influence on the viability of ram semen. In vitro fertilized two-cell stage embryos from adult sheep and prepubertal lambs, cultured in defined medium enriched with LIF, both reached the blastocyst stage at similar rates to control embryos. However, LIF exerted a positive influence on the quality of the blastocysts as revealed by significantly higher number of ICM cells and total number of cells. Together, these data demonstrate that LIF exerts a beneficial effect on sheep oocytes and embryos in vitro, but only at stages concomitant with steroid hormones surges. PMID- 16316682 TI - Multi-scale modeling to predict ligand presentation within RGD nanopatterned hydrogels. AB - The adhesion ligand RGD has been coupled to various materials to be used as tissue culture matrices or cell transplantation vehicles, and recent studies indicate that nanopatterning RGD into high-density islands alters cell adhesion, proliferation, and differentiation. However, elucidating the impact of nanopattern parameters on cellular responses has been stymied by a lack of understanding of the actual ligand presentation within these systems. We have developed a multi-scale predictive modeling approach to characterize the adhesion ligand nanopatterns within an alginate hydrogel matrix. The models predict the distribution of ligand islands, the spacing between ligands within an island and the fraction of ligands accessible for cell binding. These model predictions can be used to select pattern parameter ranges for experiments on the effects of individual parameters on cellular responses. Additionally, our technique could also be applied to other polymer systems presenting peptides or other signaling molecules. PMID- 16316681 TI - Nerve growth factor expression by PLG-mediated lipofection. AB - Biomaterials capable of efficient gene delivery provide a fundamental tool for basic and applied research models, such as promoting neural regeneration. We developed a system for the encapsulation and sustained release of plasmid DNA complexed with a cationic lipid and investigated their efficacy using in vitro models of neurite outgrowth. Sustained lipoplex release was obtained for up to 50 days, with rates controlled by the fabrication conditions. Released lipoplexes retained their activity, transfecting 48.2+/-8.3% of NIH3T3 cells with luciferase activity of 3.97x10(7)RLU/mg. Expression of nerve growth factor (NGF) was employed in two models of neurite outgrowth: PC12 and primary dorsal root ganglia (DRG) co-culture. Polymer-mediated lipofection of PC12 produced bioactive NGF, eliciting robust neurite outgrowth. An EGFP/NGF dual-expression vector identified transfected cells (GFP-positive) while neurite outgrowth verified NGF secretion. A co-culture model examined the ability of NGF secretion by an accessory cell population to stimulate DRG neurite outgrowth. Polymer-mediated transfection of HEK293T with an NGF-encoding plasmid induced outgrowth by DRG neurons. This system could be fabricated as implants or nerve guidance conduits to support cellular and tissue regeneration. Combining this physical support with the ability to locally express neurotrophic factors will potentiate regeneration in nerve injury and disease models. PMID- 16316683 TI - Oxidative degradation in highly cross-linked and conventional polyethylene after 2 years of real-time shelf aging. AB - Previous studies have reported oxidative degradation of conventional polyethylene (PE) components during shelf aging, following radiation. However, no studies have yet reported data concerning the effect of real-time shelf aging in the manufacturer's packaging on the oxidative degradation of commercially available highly cross-linking PE components. The null hypothesis tested was that in either highly cross-linked or conventional PE acetabular components there would be no significant difference in the amount of oxidative degradation between time zero PE liners and PE liners that had been real-time shelf aged for 2 years in their respective packaging. The results of the study indicated that after 2 years of real-time shelf aging, negligible oxidative degradation occurred with minimal changes in oxidation index, density, and percent crystallinity in commercially available highly cross-linked and conventional PE acetabular liners. These data suggested that oxidative degradation was not a clinical issue in the highly cross linked and conventional PE components examined after 2 years of real-time shelf aging. It is likely that current manufacturing and packaging technologies have limited the previous clinical concerns related to oxidative degradation during shelf aging of highly cross-linked and conventional PE components. PMID- 16316684 TI - In vivo dynamic MRI measurement of the noradrenaline-induced reduction in placental blood flow in mice. AB - PURPOSE: We developed a new model for in vivo placental perfusion measurements based on dynamic MRI in mice. As noradrenaline has been implicated in the pathogenesis of preeclampsia, we examined whether it reduced placental perfusion in mice, and whether such a reduction could be detected with our MRI model. MATERIALS AND METHODS: Mice at 16 days of gestation were injected intramuscularly with saline or noradrenaline solution. A conventional gadolinium chelate was then injected IV, and a single-slice T1-weighed 2D Fast SPGR sequence was acquired for 200 s. Signal intensity was measured on all the images and converted into contrast agent tissue concentrations in the maternal left ventricle (input function) and placentas. A one-compartment model was developed using compartmental and numerical modeling software. Mean blood flow (F) was calculated from a transfer constant. RESULTS: Twenty-six mice were studied, yielding a total of 55 MRI measurements of placental perfusion (29 in the control group and 26 in the noradrenaline group). Mean placental blood flow (F) was significantly lower in the noradrenaline group (0.72+/-0.84 ml/min/g of placenta) than in the control group (1.26+/-0.54 ml/min/g of placenta). CONCLUSION: Noradrenaline reduces placental perfusion in mice. Our MRI dynamic model might be useful for detecting and investigating abnormal placental blood flow, thereby avoiding the need for invasive procedures and animal sacrifice. PMID- 16316685 TI - Community- and hospital-acquired infections necessitating ICU admission: spectrum, co-morbidities and outcome. AB - OBJECTIVES: To evaluate the spectrum of infections, co-morbidities and outcome of patients admitted into an intensive care unit (ICU) due to community- (CAI) or hospital-acquired infections (HAI). METHODS: A 14-month prospective study in a medical-surgical ICU in a tertiary level teaching hospital. RESULTS: Three hundred and thirty-five patients were included, of whom 74.9% had an infection on admission; 59.3% had CAI and 40.7% HAI, while 84 patients did not have any infection (NI). The most common infections in the CAI and HAI groups (G) were pneumonia and gastrointestinal infections. Secondary bacteremia (p<0.001), severe sepsis and septic shock (p=0.048) were more prevalent in CAIG, while histories of transient ischaemic attack or stroke (p=0.03), immunosuppressive medications (p=0.009) or malignancies (p<0.001) were more common in HAIG. APACHE II scores and ICU or hospital mortalities did not differ between the groups. The median hospital stay was longer in HAIG (24 days) than in CAIG (15) or NIG (17.5), p<0.001. CONCLUSIONS: Patients in CAIG had more often secondary bacteremia or severe sepsis or septic shock, whereas HAIG patients had more often a history of cerebrovascular problems, malignancies and immunosuppressive treatments. Eighty percent of these infection patients requiring ICU treatment survived. PMID- 16316686 TI - Prevotella bivia necrobacillosis following infectious mononucleosis. AB - A case of Lemierre's syndrome is reported. Although Fusobacterium species are commonly associated with this presentation, Prevotella bivia was the causative micro-organism identified in this case. The finding that disseminated anaerobic sepsis followed primary EBV infection led to the construction of a hypothetical model of infection. PMID- 16316687 TI - Arcanobacterium haemolyticum bacteraemia and soft-tissue infections: case report and review of the literature. AB - Five cases of bacteraemia with Arcanobacterium haemolyticum in association with soft-tissue infections are reported from immunocompetent patients. A review of all A. haemolyticum isolates from our institution over a three-year period reveals that 94% were cultured from patients with soft-tissue infections, and 72% of all patients had concomitant diabetes mellitus. Arcanobacterium haemolyticum is an uncommon cause of bacteraemic soft-tissue infections, and diabetes mellitus may be an associated risk factor. PMID- 16316688 TI - An acute method for multielectrode recording from the interior of sulci and other deep brain areas. AB - Most current techniques for multielectrode recording involve chronically implanting planar or staggered arrays of electrodes. Such chronic implants are suited for studying a stable population of neurons over long periods of time but exploratory studies of the physiological properties of cortical subdivisions require the ability to sample multiple neural populations. This makes it necessary to penetrate frequently with small multielectrode assemblies. Some commercial systems allow daily penetrations with multiple electrodes, but they tend to be bulky, complex and expensive, and some make no provision for piercing the barrier of fibrous tissue that often covers the brain surface. We describe an apparatus for inserting bundles of 3-16 electrodes on a daily basis, thus allowing different neural populations to be sampled. The system is designed to allow penetration through a thick dura mater into deep brain structures. We discuss a simple method for performing multielectrode recording from cortical areas buried inside sulci using acute implantations of a bundle of electrodes. Our results show that it is possible to obtain stable recordings for at least 4h and that repeated implantations yield an average of two neurons per electrode with every electrode in the bundle picking up at least one single neuron in 70% of the implantations. PMID- 16316689 TI - Air-driven eye shutter system for vision experiments using awake behaving animals. AB - When conducting physiological and behavioral experiments for vision research, we often need to present stimuli monocularly. In this report, we describe a new eye shutter system that was developed to occlude individual eyes during experiments with awake behaving animals. The eye of choice is occluded by an air pressure driven opaque mask, the opening and closing of which is controlled by computer. This new eye shutter system has several advantages over other systems: (1) it can completely shield the eye from light using a physically opaque eye mask; (2) contamination of electrophysiological recordings by electrical noise is avoided because the actuator is not electrical; (3) the left and right eyes can be randomly occluded in each trial; (4) space for apparatus in front of the animal's face is not required. We used this eye shutter system to record neural activity from the primary visual cortex of awake monkeys and confirmed that this system is suitable for the monocular presentation of visual stimuli during electrophysiological recordings from awake animals. PMID- 16316690 TI - Transgenic Plasmodium berghei sporozoites expressing beta-galactosidase for quantification of sporozoite transmission. AB - Malaria transmission occurs during a blood-meal of an infected Anopheles mosquito. Visualization and quantification of sporozoites along the journey from the mosquito midgut, where they develop, to the vertebrate liver, their final target organ, is important for understanding many aspects of sporozoite biology. Here we describe the generation of Plasmodium berghei parasites that express the reporter gene lacZ as a stable transgene, under the control of the sporozoite specific CSP promoter. Transgenic sporozoites expressing beta-galactosidase can be simply visualized and quantified in an enzymatic assay. In addition, these sporozoites can be used to quantify sporozoites deposited in subcutaneous tissue during natural infection. PMID- 16316691 TI - Acute neutrophil activation in direct stenting: comparison of stable and unstable angina patients. AB - BACKGROUND: Polymorphonuclear neutrophils have been implicated in the pathophysiology of atherosclerosis. A substantial body of evidence has emerged to implicate the role of specific leucocyte derived enzyme myeloperoxidase in atherogenesis, since its initiation through progression until destabilization. The aim of the study was to determine the presence of polymorphonuclear neutrophils activation after coronary stenting, to compare this activation between stable and unstable setting and to evaluate the kinetic relation of this activation with inflammatory response following atherosclerotic plaque rupture. METHODS: Myeloperoxidase, lactoferrin, elastase, C-reactive protein and cytokine plasma levels were assessed in 15 patients undergoing direct coronary stenting for unstable angina (Group A) and compared to 11 patients undergoing this procedure for stable angina (Group B). Serial sampling starting before arteriography and continued for 24 h was carried out in all patients. RESULTS: A significant elevation in myeloperoxidase and lactoferrin levels was observed after stenting in both group A (p<0.0001) and group B (p<0.0001), but was higher in group A. Interleukin-8, interleukin-12 and interleukin-6 levels increased temporarily after stenting in the 2 groups. Baseline values of C-reactive protein were similar in the 2 groups and a progressive increase was observed after the intervention. CONCLUSIONS: Direct coronary artery stenting is associated with an early polymorphonuclear neutrophils activation followed by release of inflammatory cytokines (interleukin-6, interleukin-8, interleukin-12) and C reactive protein elevation in both stable and unstable patients. We conclude that stenting by itself is associated with myeloperoxidase liberation with a significantly enhanced response in unstable patients. PMID- 16316692 TI - Is an advanced age an additive risk factor to the prothrombotic or hypercoagulable state in atrial fibrillation? PMID- 16316693 TI - Cardiopulmonary exercise test evidence of isolated right coronary artery disease. AB - Isolated right coronary artery disease is usually difficult to diagnose because of frequent negativity of standard exercise stress test. We report a case of isolated coronary artery stenosis which was not detected by standard ECG stress testing. The cardiopulmonary exercise test showed a peculiar pattern: abrupt flattening in VO2/Work relationship, plateau in O2 pulse and 90 s afterwards a plateau in heart rate, probably related to vagal afferent stimulation by ischaemia of the postero-inferior wall of the left ventricle. Multidetector computed tomography of coronary vessels and coronary angiography confirmed isolated critical stenosis of middle right coronary artery. After revascularization by PTCA, normalization of cardiopulmonary exercise test was obtained. PMID- 16316694 TI - Non-dippers are associated with adverse cardiac remodeling and dysfunction (R1). AB - BACKGROUND: Non-dippers are known to carry a high risk of cardiovascular complications due to higher cumulative 24-h pressure load over time. In this study, we hypothesized that non-dippers would be associated with adverse cardiac remodeling and left ventricular (LV) dysfunction in treated hypertensive patients. MATERIALS AND METHODS: A total of 150 outpatients treated with antihypertensive drugs for at least 1 year were enrolled. The patients were classified as non-dippers if their daytime ambulatory systolic and diastolic BP did not decrease by at least 10% during the night. LV diastolic function was assessed by measuring mitral inflow velocity (E), early diastolic mitral annulus tissue velocity (E') and LV systolic function was assessed by measuring systolic tissue velocity (S'), longitudinal systolic strain and strain rate. RESULTS: The Cornell voltage product, LA volume index and LV mass index was significantly higher in non-dippers. Early diastolic mitral annulus tissue velocity (E') was lower and E/E' was higher in non-dippers suggestive of diastolic dysfunction. Systolic tissue velocity (S'), systolic strain and strain rate were significantly lower in non-dippers suggestive of systolic dysfunction. The non-dipper status, controlled for age, sex and LV mass index showed significant correlation with E' (beta=-0.203, P=0.002), E/E' (beta=0.354, P<0.001), S' (beta=-0.231, P=0.002), strain (beta=-0.162, P=0.040) and strain rate (beta=-0.186, P=0.015). CONCLUSIONS: This study showed the non-dippers in treated hypertensive patients were associated with adverse cardiac remodeling and early LV dysfunction. Further studies to demonstrate the long term prognostic significance of this finding is warranted. PMID- 16316695 TI - Home-based cardiac rehabilitation compared with centre-based rehabilitation and usual care: a systematic review and meta-analysis. AB - BACKGROUND: To determine the effectiveness of home-based cardiac rehabilitation programmes compared with (i) usual care and (ii) supervised centre-based cardiac rehabilitation on mortality, health related quality of life and modifiable cardiac risk factors of patients with coronary heart disease. METHODS: Systematic review and meta-analysis of randomised controlled trials. MAIN OUTCOME MEASURES: mortality, smoking cessation, exercise capacity, systolic blood pressure, total cholesterol, psychological status, and health related quality of life. RESULTS: Eighteen included trials for home versus usual rehabilitation and six trials of home versus supervised centre-based rehabilitation were identified. The home based interventions were clinically heterogeneous, trials often small, with quality poorly reported. Compared with usual care, home-based cardiac rehabilitation had a 4 mm Hg (95% CI 6.5, 1.5) greater reduction in systolic blood pressure, and a reduced relative risk of being a smoker at follow-up (RR 0.71, 95% CI 0.51, 1.00). Differences in exercise capacity, total cholesterol, anxiety and depression were all in favour of the home-based group. In patients post-myocardial infarction exercise capacity was significantly improved in the home rehabilitation group by 1.1 METS (95% CI 0.2, 2.1) compared to usual care. The comparison of home-based with supervised centre-based cardiac rehabilitation revealed no significant differences in exercise capacity, systolic blood pressure and total cholesterol. CONCLUSIONS: Current evidence does not show home-based cardiac rehabilitation to be significantly inferior to centre-based rehabilitation for low-risk cardiac patients. However, the numbers of patients included are less than 750 and ongoing trials will contribute to the debate on the acceptability, effectiveness and cost-effectiveness of home-based cardiac rehabilitation. PMID- 16316696 TI - Elevated matrix metalloproteinase expression after stent implantation is associated with restenosis. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) play a key role in intimal growth and is responsible for ventricular remodeling after stent implantation. However, little is known about the relationship between early MMPs expression post-stent implantation and follow-up restenosis. METHODS: We investigated the serial changes of serum MMP-9, MMP-2 and tissue inhibitor of metalloproteinase-1 (TIMP 1) in 16 control subjects with normal coronary angiography (control) and 40 patients before and on the 1st, 3rd and 7th day after uncomplicated stent implantation. Follow-up angiography was performed at 6 months after stent implantation. RESULTS: Serum MMP-2 level was higher in patients with restenosis on the 1st day post-stent implantation and returned to pre-operation level thereafter. Serum MMP-9 levels consistently increased in patients with restenosis up to 7th day post-stent implantation; MMP-9 levels in the 1st, 3rd and 7th day after stent implantation were positively correlated to the late loss index 6 months after stent implantation. CONCLUSIONS: Increased serum MMP-9 level is associated with increased risk of restenosis post-stent implantation. PMID- 16316697 TI - Intermittent right axis deviation in the presence of complete left bundle branch block. PMID- 16316698 TI - Primary impairment of left ventricular function in Marfan syndrome. AB - BACKGROUND: Cardiovascular involvement in Marfan syndrome is mainly characterized by progressive dilatation of the proximal aorta. Whether left ventricular dysfunction is present in these patients is not clear at present. OBJECTIVES: Assess left ventricular function in patients with Marfan syndrome, free of significant valvular heart disease, using a combination of MRI and Tissue Doppler imaging (TDI). METHODS AND RESULTS: A total of 26 Marfan patients (mean age=32.0+/-10.9, 12 men) without significant valvular heart disease, and 26 age- and sex-matched controls were studied. Left ventricular volumes and ejection fraction were measured with magnetic resonance imaging. Systolic and diastolic function parameters were assessed using conventional echocardiography and TDI. When compared to controls, Marfan patients showed impairment of left ventricular contractile function as expressed by a reduced ejection fraction (53.5+/-9.0% vs. 59.6+/-6.7%, p=0.009), an increased end-systolic volume (36.0+/-9.5 vs. 29.5+/ 6.7 ml/m(2), p=0.007), and reduced peak systolic velocities at the basal septal and lateral myocardial wall (5.2+/-1.4 vs. 6.4+/-1.3 cm/s, p=0.003 and 6.0+/-2.2 vs. 7.5+/-2.3 cm/s, p=0.03, respectively). Diastolic function was impaired with an increased deceleration time of the E wave (171+/-41 ms vs. 141+/-36 ms, p=0.006). Peak early diastolic velocity at the mitral valve annulus was significantly lower (9.6+/-2.4 cm/s vs. 11.9+/-3.3 cm/s, p=0.006). CONCLUSION: These data provide evidence for mild, but significant impairment of left ventricular systolic and diastolic function in Marfan patients, not related to valvular heart disease. PMID- 16316699 TI - Coronary risk factors and inflammation in patients with coronary artery disease and internal cardioverter defibrillator implants. AB - BACKGROUND: The internal cardioverter defibrillator (ICD) is increasingly used to treat ventricular tachyarrhythmias in patients with coronary artery disease (CAD). The burden of coronary risk factors and inflammation is however not well studied in these high risk patients. STUDY AIMS: The aim of the present study was to describe the prevalence of coronary risk factors (including lipid values) and inflammation (including high sensitive-C-reactive protein, hs-CRP) in patients with CAD and ICD implants. METHODS: Baseline clinical characteristics and laboratory results of all eligible patients for the Cholesterol Lowering and Arrhythmias Recurrences after Internal Defibrillator Implantation trial (CLARIDI trial) were used. All patients had documented CAD, an ICD implant and were not yet treated with statins. Coronary risk factors, lipid values, glycated haemoglobin (HbA(1c)) and hs-CRP levels were determined. RESULTS: In the 110 included patients (mean age 68+/-9 years, LVEF 40+/-17%, NYHA class II-III in 47%), a high prevalence of coronary risk factors was documented: current smoking in 18%, body mass index > or =30 kg/m(2) in 16%, blood pressure > or =140/90 mm Hg in 40%, history of diabetes in 12%, and HbA(1c) > or =6% in 16% of patients not known with diabetes. A total cholesterol >175 mg/dl was found in 76% of patients and an LDL cholesterol >100 mg/dl in 83%. Finally, median hs-CRP was 4.8 mg/l (interquartile range 2.5-13.9 mg/l). Hs-CRP values > or =2 mg/l were noted in 83% of all patients and in 68% of patients who had an ICD implant more than 6 months before inclusion. CONCLUSION: In CAD patients with ICD implants, the burden of coronary risk factors is high, often unrecognized and/or under-treated. Persistent inflammation is found in the majority of these patients. PMID- 16316700 TI - Development of a semi-automated colorimetric assay for screening anti-leishmanial agents. AB - MTS or {3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl} 2H-tetrazolium, inner salt) is converted into soluble formazan by mitochondrial dehydrogenase of viable cells, thus serving as an indicator of cell viability. Accordingly, a MTS-based assay was developed to evaluate anti-leishmanial activity in Leishmania promastigotes from strains responsible for visceral, cutaneous or mucocutaneous leishmaniasis. The assay was initially optimized for the appropriate wavelength (490 nm), culture medium (M-199), incubation time (3 h) and temperature (37 degrees C). Increasing absorbance with increasing cell density confirmed linearity of the assay that was maintained up to 2.5 x 10(6) cells/200 microl. The growth kinetics of six L. donovani strains and six non-L. donovani strains consistently indicated higher absorbances in the L. donovani strains highlighting the importance of strain-specific customization of the MTS assay. The IC(50) values (i.e., the concentration at which 50% of growth was inhibited) of amphotericin B, miltefosine and pentamidine isethionate obtained by the MTS assay corroborated with previously published data. Taken together, the MTS assay thus permits a simple, reproducible and reliable semi-automated method for evaluating cell viability, effective for drug-screening and growth kinetic studies. PMID- 16316701 TI - Immobilization of hydrocarbon-oxidizing bacteria in poly(vinyl alcohol) cryogels hydrophobized using a biosurfactant. AB - A simple biosurfactant-based hydrophobization procedure for poly(vinyl alcohol) (PVA) cryogels was developed allowing effective immobilization of hydrocarbon oxidizing bacteria. The resulting partially hydrophobized PVA cryogel granules (granule volume 5 microl) contained sufficient number (6.5 x 10(3)) of viable bacterial cells per granule, possessed high mechanical strength and spontaneously located at the interface in water-hydrocarbon system. Such interfacial location of PVA granules allowed high contact of immobilized biocatalyst with hydrophobic substrate and water phase, thus providing bacterial cells with mineral and organic nutrients. As a result, n-hexadecane oxidation efficiency of 51% after 10 day incubation was achieved using immobilized biocatalyst. PVA cryogels with increased hydrophobicity can be used for immobilization of bacterial cultures performing oxidative transformations of water-immiscible organic compounds. Immobilization of in situ biosurfactant producing Rhodococcus bacteria into PVA cryogel is discussed. PVA cryogel granules with entrapped alkanotrophic rhodococcal cells were stable after 10-month storage at room temperature. PMID- 16316702 TI - Monitoring changes in nisin susceptibility of Listeria monocytogenes Scott A as an indicator of growth phase using FACS. AB - Listeria monocytogenes has previously been shown to adapt to a wide variety of environmental niches, principally those associated with low pH, and this compromises its control in food environments. An understanding of the mechanism(s) by which L. monocytogenes survives unfavourable environmental conditions will aid in developing new food processing methods to control the organism in foodstuffs. The present study aimed to gain a further understanding of the physiological basis for the differential effects of one control strategy, namely the use of the lantibiotic nisin. Using propidium iodide (PI) to probe membrane integrity it was shown that L. monocytogenes Scott A was sensitive to nisin (8 ng mL(-)) but this was growth phase dependent with stationary phase cells (OD600=1.2) being much more resistant than exponential phase cells (OD600=0.38). We demonstrate that, using a combination of techniques including fluorescence activated cell sorting (FACS), the membrane adaptations underpinning nisin resistance are triggered much earlier (OD600<0.5) than the onset of stationary phase. The significance of these findings in terms of mechanism and application are discussed. PMID- 16316703 TI - Molecular tools to detect the IncJ elements: a family of integrating, antibiotic resistant mobile genetic elements. AB - The IncJ group of enterobacterial mobile genetic elements, which include R391, R392, R705, R997 and pMERPH, have been shown to be site-specific integrating elements encoding variable antibiotic and heavy metal resistance genes. They insert into a specific 17-bp site located in the prfC gene, encoding peptide release factor 3, in Escherichia coli and other hosts. A key feature of known IncJ elements is the presence of a site-specific recombination module consisting of an attachment site on the element and an integrase-encoding gene of the tyrosine recombinase class, which promotes integration between the attachment site on the element and a similar site on the host chromosome. We have cloned and sequenced the integrases from a number of known IncJ elements and designed PCR primers for specific amplification of this gene. Using conserved regions of enterobacterial prfC genes upstream and downstream of the insertion site, and conserved sequences at the ends of the integrated IncJ elements, we have designed specific primers to amplify across the integrated IncJ attL and attR junction fragments. Alignment of over 30 enterobacterial prfC-like genes indicates that the primers designed to amplify attR junction would amplify IncJ element: host junctions from a wide variety of hosts. The IncJ elements have been shown to sensitise recA(+)E. coli K12 strains to UV irradiation. A simple and rapid procedure for demonstrating this effect is described. These tools should enable the rapid detection of such elements in clinical and environmental settings. PMID- 16316704 TI - Occurrence and daily intake of ochratoxin A of organic and non-organic rice and rice products. AB - Ochratoxin A (OTA) was extracted from 84 rice samples and rice products by using accelerated solvent extraction (ASE) and analysed with liquid chromatography coupled with fluorescence detection. Samples were collected from rice cultivars, local markets and supermarkets; 64 were of non-organic and 20 of organic production. 7.8% of non-organic samples had OTA levels from 4.3 to 27.3 microg/kg and in 30% of organic samples was detected the presence of this mycotoxin varying from 1.0 to 7.1 microg/kg. OTA presence was confirmed by methyl-ester derivatization. Rice and rice products labelled with denomination of origin (DO) were not detected OTA due to the fact that its production has implemented food safety measures such as good agricultural practices (GAPs), good manufacturing practices (GMPs), and the hazard analysis and critical control point (HACCP) system. Estimated daily intake of OTA was 0.17 ng/kg b.w./day. This value reflects that the analysed samples have a minimal contribution to toxicological risk. PMID- 16316705 TI - Liver targeting of catalase by cationization for prevention of acute liver failure in mice. AB - To achieve hepatic delivery of CAT for the prevention of CCl4-induced acute liver failure in mice, two types of cationized CAT derivatives, HMD- and ED-conjugated CAT, were developed. Slight structural changes occurred during cationization and the number of increased free amino groups was 3.1 in HMD-CAT and 13.6 in ED-CAT. 111In-cationized CAT derivatives showed an increased binding to HepG2 cells, and were rapidly taken up by the liver. H2O2-induced cytotoxicity in HepG2 cells was significantly prevented by preincubation of the cells with cationized CAT derivatives. A bolus intravenous injection of the cationized CAT derivatives reduced the hepatotoxicity induced by CCl4 in mice. The ED-CAT, which showed more rapid and greater binding to the liver than the HMD-CAT, exhibited more beneficial effects as far as all the parameters examined (serum GOT, GPT, LDH and hepatic GSH) were concerned, suggesting that a high degree of cationization is effective in delivering CAT to the liver to prevent CCl4-induced hepatotoxicity. These results suggest that cationized CAT derivatives are effective in preventing acute liver failure, and ED-based cationization is a suitable method for developing liver-targetable cationized CAT derivatives, because it provides CAT with a high degree of cationization and a high remaining enzymatic activity. PMID- 16316706 TI - Regulatory aspects of drug approval for macular degeneration. AB - Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries with approximately 15 million people with the disease in the United States. AMD is characterized as a progressive degenerative disease of the macula. There are two forms of AMD: neovascular and non-neovascular. The non neovascular form of AMD is more common and leads to a slow deterioration of the macula with a gradual loss of vision over a period of years. The neovascular form of the disease is responsible for the majority of cases of severe vision loss and is due to proliferation of abnormal blood vessels behind the retina. These blood vessels leak blood and fluid into the retina, which results in visual abnormalities. The development of these abnormal blood vessels is due in part to the activity of VEGF (vascular endothelial growth factor) and its inhibition is expected to impact on the onset and/or severity of vision loss associated with the proliferation of abnormal blood vessels. Age-related macular degeneration is an underserved treatment population. Visudyne (verteporfin for injection), Novartis Ophthalmics, is FDA approved for the treatment of patients with predominantly classic subfoveal choroidal neovascularization due to age-related macular degeneration, pathologic myopia or presumed ocular histoplasmosis. Pegaptanib sodium (Macugen, Eyetech Pharmaceuticals, Inc. and Pfizer, Inc.) is indicated for the treatment of neovascular (wet) age-related macular degeneration. This article will review the approval pathway for these two products and that required of future products indicated for macular degeneration. PMID- 16316707 TI - Complete resection in lung cancer surgery: proposed definition by R. Rami-Porta, C. Wittekind and P. Goldstraw. PMID- 16316708 TI - Neuroprotection by tetramethylpyrazine against ischemic brain injury in rats. AB - In traditional Chinese medicine, Ligusticum wallichii Franchat (Chuan Xiong) and its active ingredient tetramethylpyrazine (TMP) have been used to treat cardiovascular diseases and to relieve various neurological symptoms such as ischemic deficits. However, scientific evidence related to their effectiveness or precise modes of neuroprotective action is largely unclear. In the current study, we elicited the neuroprotective mechanisms of TMP after focal cerebral ischemic/reperfusion (I/R) by common carotid arteries and middle cerebral artery occlusion model in rats. TMP was administrated 60 min before occlusion via intraperitoneal injection. TMP concentration-dependently exhibited significant neuroprotective effect against ischemic deficits by reduction of behavioral disturbance. Neuronal loss and brain infarction in the ischemic side of rats were markedly lowered by treatment with TMP. Cerebral I/R-induced internucleosomal DNA fragmentation, caspase-8, caspase-9, and caspase-3 activation, and cytochrome c release were reduced by TMP treatment. Western blot analysis revealed the down regulation of Bcl-2 and Bcl-xL and the up-regulation of Bax and Bad by cerebral I/R insult. Among them, only the alteration in Bcl-xL expression was reversed by TMP treatment. Moreover, the activation of microglia and/or recruitment of inflammatory cells within the ischemic side and the consequent production of monocyte chemoattractant protein 1 (MCP-1) were suppressed by TMP pre-treatment. Our findings suggest that TMP might provide neuroprotection against ischemic brain injury, in part, through suppression of inflammatory reaction, reduction of neuronal apoptosis, and prevention of neuronal loss. PMID- 16316709 TI - Smad-dependent alterations of PPT cholinergic neurons as a pathophysiological mechanism of age-related sleep-dependent memory impairments. AB - In humans, memory impairments are highly prevalent in the aged population, but their functional and structural origins are still unknown. We hypothesized that circadian rhythm alterations may predict spatial memory impairment in aged rats. We demonstrate an association between sleep/wake circadian rhythm disturbances (non-REM sleep fragmentation) and spatial memory impairments in aged rats. We show by light and electron microscopy that these age-related disruptions in circadian rhythm and spatial memory are also associated with degeneration of cholinergic neurons of the pedunculopontine nucleus (PPT), a structure known to be involved in sleep and cognitive functions and which is altered during aging. Finally, we demonstrate that a trophic deregulation of the PPT occur in aged impaired rats, involving an over activation of the TGFbeta-Smad cascade, a signalling pathway involved in neurodegeneration. In conclusion these results provide a new pathophysiological mechanism for age-related sleep-dependent memory impairments opening the ground for the development of new therapeutic approaches of these pathologies. PMID- 16316710 TI - Very small size proteoliposomes derived from Neisseria meningitidis: An effective adjuvant for generation of CTL responses to peptide and protein antigens. AB - The development of potent adjuvants, conditioning innate and adaptative immunity, particularly CTL responses, has become currently a hot point in the rational design of vaccines for cancer immunotherapy. We have described a new approach, in which gangliosides are incorporated into vesicles from Neisseria meningitidis to form Very Small Size Proteoliposomes (VSSP). VSSP is a good alternative to the existing adjuvants for use in whole cells vaccines since it promotes 80% tumour rejection and growing delay in the CT26 and F3II tumour models respectively. Also VSSP induces activation of CTL responses to co-injected trimmed peptides and soluble proteins. This phenomena is facilitated by the cross-presentation of exogenous antigen and do not need cooperation of CD4 T cells for primary CD8 T cells expansion. PMID- 16316711 TI - DNA vaccines for therapy of tuberculosis: where are we now? AB - DNA vaccines that were being investigated in mice for prophylactic use against tuberculosis were soon found also to be surprisingly effective as treatment against established infection. The immune system was stimulated to kill the bacteria, even including the persistent latent bacteria that are otherwise refractory to the immune system and antibacterial chemotherapeutic drugs alike. Subsequent results from a range of laboratories using diverse DNA vaccines in diverse murine models of infection have been very varied, ranging from enhanced pathology, through negligible effects, to major additive benefit from combined vaccine and chemotherapy. This review summarises the data and assesses future prospects. PMID- 16316712 TI - The vaccine potential of Streptococcus pneumoniae surface lectin- and non-lectin proteins. AB - We have fractionated S. pneumoniae surface proteins into lectin and non-lectin fractions and tested their ability to elicit protective immune responses in the mouse model system. The total cell wall protein fraction (CW-T) was separated into lectin (CW-L), and non-lectin (CW-NL) fractions and used for immunization of mice. Immunized mice were challenged intranasally or intraperitoneally with S. pneumoniae strain WU2 (serotype 3). CW-T, CW-NL and CW-L and adjuvant only vaccination protected 55%, 43%, 44% and 0% of the intranasally challenged mice, respectively and 67%, 86%, 11% and 0% of mice challenged intraperitoneally, respectively. Immunogenic proteins in each fraction were sequenced and identified using MALDI-TOF. CW-L proteins provided a significantly better protection against intranasal inoculation and CW-NL proteins provided a significantly better protection from intraperitoneal inoculation. Proteins identified by sera from mice immunized with the cell-wall derived fractions may constitute candidates for future development of anti S. pneumoniae vaccines. PMID- 16316713 TI - Dengue tetravalent DNA vaccine inducing neutralizing antibody and anamnestic responses to four serotypes in mice. AB - We developed a dengue tetravalent DNA vaccine consisting of plasmids expressing premembrane and envelope genes of each of four serotypes of dengue viruses. BALB/c mice immunized twice with the tetravalent vaccine at a dose of 100 microg (25 microg for each serotype) using a needle-free jet injector developed neutralizing antibodies against all serotypes. There was no interference among the four components included in this combination vaccine. Tetravalent vaccine immunized mice showed anamnestic neutralizing antibody responses following challenge with each dengue serotype: responses to challenges from serotypes different to those used for neutralization tests were also induced. PMID- 16316714 TI - Recasting research into children's experiences of parental mental illness: beyond risk and resilience. AB - Children who live with a mentally ill parent are viewed primarily as being 'at risk' of developing a mental illness themselves and those who remain well are considered extraordinarily resilient. This particular risk/resilience discourse is embedded within larger contemporary discourses about risk and childhood. Childhood is seen as a critical period of development during which children need protection due to their physical and psychological vulnerabilities. In this paper, the implications of this dominant casting of children are explored and it is argued that the conceptual repertoire about those living with a mentally ill parent should be expanded. A critique of the literature that established the risk/resilience discourse is followed by a discussion of research about parenting with a mental illness within which children are surprisingly absent. Recent thinking about children arising out of the 'new' social studies of childhood is summarized to illustrate its resistance to the hegemonic image of children as passive, developing, 'unfinished' persons. A recasting of children as complex young persons who have competencies as well as vulnerabilities linked to their developmental stages, would lead to different lines of inquiry about children's experiences of mental illness in a parent. PMID- 16316715 TI - Effect of gender on antinociceptive effect of paroxetine in hot plate test in mice. AB - PURPOSE: Many studies have reported both a gender difference in the rates of depression and its treatment by using any of the widely used antidepressant drug groups. Some studies suggest that females respond more poorly to tricyclic antidepressants than males and appear to respond better to selective serotonin reuptake inhibitors (SSRI). There is no study investigating the analgesic/antinociceptive effects of antidepressant drugs on the basis of gender difference. In this study, we aimed to investigate the antinociceptive effect of paroxetine on the basis of gender difference. METHODS: The antinociceptive effect of paroxetine was tested using hot plate test in Balb/c mice (30-40 g). The animals were divided into eight groups on the basis of gender. FINDINGS: While paroxetine did not induce an antinociceptive effect in both sex at a dose of 1 mg kg(-1), it showed significant antinociceptive effects in both sex at a dose of 5 or 10 mg kg(-1). None of the doses of paroxetine revealed a gender difference in its antinociceptive action. CONCLUSION: There are several studies showing positive or negative evidence on the gender difference of paroxetine's antidepressant effect, but in the literature there is no study about the gender difference of paroxetine's or any other SSRI drug's antinociceptive effect. In conclusion, our results do not show any gender difference in antinociceptive effect of paroxetine that may be important especially when it would be used as an adjuvant agent in some painful conditions. PMID- 16316716 TI - Plasma clozapine levels and clinical response in treatment-refractory Chinese schizophrenic patients. AB - PURPOSE: To evaluate clinical efficacy of clozapine in relation with its plasma level in a group of Chinese patients with treatment-resistant schizophrenia. In addition, the relationship between plasma level and side effects were examined. METHOD: Fifty-one patients with treatment-resistant schizophrenia were put on a fixed dose of clozapine at 300 mg/day for 6 weeks. Non-responders to week 6 received 500 mg/day in subsequent 6 weeks. Responders to week 6 continued to receive 300 mg/day. Clozapine plasma levels were checked at weeks 6 and 12. FINDINGS: No association was found between clozapine plasma level, response and side effects. Sodium valproate was found to elevate clozapine plasma level while lowering norclozapine/clozapine ratio. CONCLUSION: Clozapine plasma level was not found to be associated with response and side effect in Chinese treatment resistant schizophrenic patients. Various explanations were postulated for the lack of relationship observed between clozapine plasma level and response in this population. PMID- 16316717 TI - A capability model of individual differences in frontal EEG asymmetry. AB - Researchers interested in measuring individual differences in affective style via asymmetries in frontal brain activity have depended almost exclusively upon the resting state for EEG recording. This reflects an implicit conceptualization of affective style as a response predisposition that is manifest in frontal EEG asymmetry, with the goal to describe individuals in terms of their general approach or withdrawal tendencies. Alternatively, the response capability conceptualization seeks to identify individual capabilities for approach versus withdrawal responses during emotionally salient events. The capability approach confers a variety of advantages to the study of affective style and personality, and suggests new possibilities for the approach/withdrawal motivational model of frontal EEG asymmetry and emotion. Logical as well as empirical arguments supportive of this conclusion are presented. PMID- 16316718 TI - Biochemical hypothyroidism--a new finding in mirror syndrome? PMID- 16316719 TI - Metabolic response to exogenous ethanol in yeast: an in vivo NMR and mathematical modelling approach. AB - The understanding of the metabolic behaviour of complex systems such as eukaryotic cells needs the development of new approaches that are able to deal with the complexity due to a large number of interactions within the system. In this paper, we applied an approach based on the combined use of in vivo NMR experiments and mathematical modelling in order to analyze the metabolic response to ethanol stress in a wild-strain of Saccharomyces cerevisiae. Considering the cellular metabolic processes resulting from activation, inhibition, and feed-back activities, we developed a model able to describe the modulation of the whole system induced by an external stress due to increasing concentrations of exogenous ethanol. This approach was able to interpret the experimental results in terms of metabolic response to exogenous ethanol in the yeast. The robustness and flexibility of the model enables it to work correctly at different initial exogenous ethanol concentrations. PMID- 16316720 TI - Vascular cognitive impairment in patients with late-onset seizures after an ischemic stroke. AB - BACKGROUND: Cognitive impairment and seizures are both common conditions in patients with cerebrovascular disease. PURPOSE: The present study investigates whether the occurrence of late-onset seizures, following an ischemic stroke, contributes to vascular cognitive impairment. PATIENTS AND METHODS: The mean Mini Mental State Examination (MMSE) and the median modified Rankin (mR) scores were compared between 125 patients who developed late-onset seizures (66 with a single seizure and 59 with repeated seizures or epilepsy) following an ischemic stroke and 125 patients who did not during, at least, a 2-year follow-up. RESULTS: There were no differences in age, gender, etiology and degree of neurological impairment on admission for their stroke between the groups with and without seizures. Although the mean MMSE score was similar between both groups the median mR score was significantly higher in the seizure patients. Comparing the patients with a single seizure to the non-seizure ones showed the same results. On the other hand, comparison of the patients with epilepsy to the non-seizure group revealed, in addition to the higher median mR score, a significantly lower mean MMSE score in the former group. CONCLUSION: Repeated seizures following an ischemic stroke promote vascular cognitive impairment. PMID- 16316721 TI - Triptolide induces Bcl-2 cleavage and mitochondria dependent apoptosis in p53 deficient HL-60 cells. AB - Triptolide, a bioactive component of the Chinese medicinal herb Tripterygium wilfordii Hook F., induces p53-mediated apoptosis in cancer cells. This study demonstrated that triptolide activated an alternative p53-independent apoptotic pathway in HL-60 cells. In the absence of an intact p53 and without changing Bax level, at nM range triptolide induced apoptosis with concomitant DNA fragmentation, S phase cell cycle arrest, mitochondrial cytochrome c release and the activation of caspases. Besides, both caspases 8 and 9 were activated and the simultaneous inhibition of both was required to completely block triptolide's apoptotic effect. Importantly, triptolide induced the appearance of a truncated 23kD Bcl-2 which was inhibited by the general caspase inhibitor Z-VAD-FMK. In the MCF-7 cells that possessed the wild type p53 but lacked caspases 3, triptolide induced cell death with an increase in p53 but Bcl-2 remained unaltered. On the other hand, transfected cells overexpressing the 28kD Bcl-2 became more resistant to triptolide and upon triptolide treatment accumulated in the G(1) instead of S phase. After 36h treatment, triptolide activated JNK pathways, at the same time inactivated the ERK and p38 pathways. However, SP600125, a specific JNK inhibitor, could not inhibit the triptolide-mediated cleavage of caspase 3, indicated that activation of JNK might not be related to the apoptotic effects of triptolide. Our data suggest that in the absence of an intact p53 and without altering Bax level triptolide induces apoptosis activates a positive amplification loop involving caspase-mediated Bcl-2 cleavage/activation, mitochondrial cytochrome c release and further activation of caspases. PMID- 16316722 TI - Myxoid leiomyosarcoma of the liver. AB - This is the first report of a myxoid leiomyosarcoma arising in a cirrhotic liver. The tumor was resected from a 64-year-old man. On gross examination, it was soft and hemorrhagic. The tumor was composed of deceptively benign-looking smooth muscle cells with clear cytoplasm suspended in a myxoid stroma with foci of hemorrhage. Immunohistochemistry and electron microscopy confirmed that this was a smooth muscle cell neoplasm. The abundance of glycogen and ultrastructural signs of smooth muscle differentiation were considered consistent with an immature smooth muscle cell phenotype consistent with the diagnosis of myxoid leiomyosarcoma. Since myxoid leiomyosarcomas are aggressive tumors, it is important to recognize them histologically and also bear in mind that these tumors can occur even in unusual extrauterine locations such as a cirrhotic liver. PMID- 16316723 TI - Chronic ecstasy (MDMA) use is associated with deficits in task-switching but not inhibition or memory updating executive functions. AB - The study aimed to establish whether prior ecstasy (MDMA) use caused impairments in tasks which targeted specific executive functions and whether the magnitude of impairments were correlated with the level of drug use. Subjects who had used ecstasy and cannabis (N=33), subjects who had used cannabis but not ecstasy (N=17) and subjects who had used neither drug (N=18) were tested on a standard and modified form of the Stroop Colour-Word test to look for impairments in inhibition and in task-switching, respectively. A Keep Track Task was used to assess working memory updating ability. Ecstasy users, but neither of the other groups were significantly impaired (slower reaction times) on the task-switching version of the Stroop test (p<.05), but not on the standard version or on the Keep Track Task. Regression analysis showed a significant correlation between level of lifetime ecstasy use and magnitude of the switching-impairment. The findings are interpreted in line with recent theoretical views which regard executive function as composed of multiple independent cognitive components such as inhibition, switching and memory updating, only some of which may be affected by drug use or other factors. PMID- 16316724 TI - Alteration of RPL14 in squamous cell carcinomas and preneoplastic lesions of the esophagus. AB - Allelic loss on chromosome 3p occurs frequently in esophageal cancer. The human ribosomal protein L14 gene (RPL14) is located on chromosome 3p21.3. In the present study, we investigated alteration of RPL14 at both the genomic DNA and RNA levels in 129 Chinese esophageal squamous cell carcinomas (ESCC) and 17 dysplasia adjacent to tumor tissues by a combination of tissue microdissection, microsatellite analysis of the intragenic marker, reverse transcriptase polymerase chain reaction (RT-PCR) and direct sequencing. In the tested informative cases, loss of heterozygosity (LOH) of RPL14 was observed in 29 out of 68 (43%) tumors. Decreased expression of the gene was detected in 31 out of 49 (63%) carcinomas. No mutation was found in the remaining RPL14 allele of the tumors with LOH. We examined subsequently the allelic status of RPL14 in the dysplasia (preneoplastic lesions) between malignant tissues and histologically normal epithelia. Of 17 tested dysplasia in which the tumors showed LOH, eight (47%) displayed the same allelic loss as their corresponding tumors, seven (41%) exhibited microsatellite instability (MSI), and only two retained both the RPL14 alleles. The data suggest that alteration of RPL14 occurred frequently in ESCC and might be an earlier event in the tumorigenesis of the esophagus. Analysis to RPL14 gene may contribute to the early detection of ESCC as a potential molecular marker. PMID- 16316725 TI - An intronic polyadenylation site in human and mouse CstF-77 genes suggests an evolutionarily conserved regulatory mechanism. AB - Human CstF-77 is one of the three subunits of cleavage stimulation factor (CstF) that is essential for mRNA polyadenylation. Its Drosophila homologue, suppressor of forked [su(f)], contains an intronic poly(A) site, which can lead to a short transcript without a stop codon. By both bioinformatic searches and validation with molecular biology experiments, we found that human and mouse CstF-77 genes also contain an intronic poly(A) site, which can be utilized to produce short CstF-77 transcripts lacking sequences encoding domains that are involved in many of the CstF-77 functions. The genomic sequence surrounding the poly(A) site is highly conserved among all vertebrates, but is not present in non-vertebrate species. Using public Serial Analysis of Gene Expression (SAGE) data, we found that the intronic poly(A) site is utilized in a wide range of tissues. This finding indicates that vertebrates may employ a similar alternative polyadenylation mechanism to modulate CstF-77, highlighting the importance of the regulation of CstF-77 in various species. PMID- 16316726 TI - Tenebrio molitor antifreeze protein gene identification and regulation. AB - The yellow mealworm, Tenebrio molitor, is a freeze susceptible, stored product pest. Its winter survival is facilitated by the accumulation of antifreeze proteins (AFPs), encoded by a small gene family. We have now isolated 11 different AFP genomic clones from 3 genomic libraries. All the clones had a single coding sequence, with no evidence of intervening sequences. Three genomic clones were further characterized. All have putative TATA box sequences upstream of the coding regions and multiple potential poly(A) signal sequences downstream of the coding regions. A TmAFP regulatory region, B1037, conferred transcriptional activity when ligated to a luciferase reporter sequence and after transfection into an insect cell line. A 143 bp core promoter including a TATA box sequence was identified. Its promoter activity was increased 4.4 times by inserting an exotic 245 bp intron into the construct, similar to the enhancement of transgenic expression seen in several other systems. The addition of a duplication of the first 120 bp sequence from the 143 bp core promoter decreased promoter activity by half. Although putative hormonal response sequences were identified, none of the five hormones tested enhanced reporter activity. These studies on the mechanisms of AFP transcriptional control are important for the consideration of any transfer of freeze-resistance phenotypes to beneficial hosts. PMID- 16316727 TI - A new scrambled gene in the ciliate Uroleptus. AB - In the germline micronucleus of spirotrichous ciliates, the gene segments, or macronuclear destined sequences (MDSs), that give rise to the somatic macronucleus are interrupted by internal eliminated sequences (IESs). For some genes, the MDSs are not arranged sequentially, but rather are scrambled, in the micronucleus. Three scrambled genes have been extensively studied in many species: actin I, alpha-telomere binding protein, and DNA polymerase alpha. However, in the past decade, no new scrambled genes have been reported, and the prevalence of scrambled genes is still an important question. To screen for scrambled genes, we completely sequenced 11 macronuclear chromosomes in the spirotrich Uroleptus sp., and then pursued their micronuclear organization. This allowed us to identify new scrambled genes, which also display novel features. In this study we describe one of these newly discovered scrambled genes. This gene, tentatively named USG1 (Unknown Scrambled Gene 1), encodes a putative protein of 1016 aa. While the function of this protein product is not clear, dN/dS calculated from the two alleles suggests the encoded protein is under purifying selection. USG1 consists of 16 germline MDSs, of which 14 are located on one locus. The other locus, which is at least 3 kb away from the main locus, contains two scrambled MDSs separated by a nonscrambled IES. Curiously, one MDS and its outgoing (3') pointer (direct repeat) overlap intron splice sites, indicating that these DNA sequences may be under dual (or multiple) constraints. Our findings identify a new scrambled gene in the micronuclear genome of a spirotrichous ciliate, and suggest that even more complicated structures may be present. PMID- 16316728 TI - The molecular epidemiology of Mycobacterium bovis infections in Tanzania. AB - A molecular epidemiological study to determine the zoonotic importance of bovine tuberculosis was carried out in Tanzania. Specimens from human cases of tuberculosis as well as from slaughtered cattle were collected from regions with a high proportion of extrapulmonary tuberculosis. In order to determine the similarity of strains from the two sources, molecular typing techniques, namely RFLP and spoligotyping, were used to determine the genetic profile of the strains involved. The results of pTBN12 typing of M. bovis from cattle and man has shown a rather heterogeneous population of this species spread all over Tanzania, assuming that the present sample is representative. There were 13 different pTBN12 RFLP types encountered. The genetic relatedness between the pTBN12 RFLP patterns indicated a high degree of relatedness (86%) between the dominant pTBN12 genotypes existing in Tanzania. There were 13 different spoligotypes found in this study, whose genetic relatedness was also high (79%). DNA profiles were also confirmed by IS986 RFLP, which revealed that strains have 1-13 copies of IS986. Geographically, there was overlap between pTBN12 RFLP and spoligotypes amongst strains isolated from various parts of Tanzania. The diversity of the RFLP and spoligotype patterns observed in Tanzania probably reflects the extensive internal movements of cattle belonging to pastoralists. The evidence of overlap between DNA fingerprints of M. bovis from cattle and man has once more highlighted a need for synergy of veterinary and medical policies in the control of tuberculosis in Tanzania and probably in other developing countries. PMID- 16316729 TI - Mitoxantrone-loaded BSA nanospheres and chitosan nanospheres for local injection against breast cancer and its lymph node metastases. I: Formulation and in vitro characterization. AB - Positively charged mitoxantrone (MTO) was absorbed by negatively charged blank bovine serum albumin (BSA) and chitosan (CS) nanospheres to form MTO-BSA-NS and MTO-CS-NS, respectively. In addition to other conditions, values of pH of every step were optimized. On optimized conditions, MTO-BSA-NS of a mean size of 77 nm with an encapsulation yield (EY) of (98.86+/-1.43)% [drug loading rate (DL) (19.82+/-0.29)%] and MTO-CS-NS of a mean size of 75 nm with an EY of (97.57+/ 1.00)% [DL (9.78+/-0.10)%] were obtained. After lyophilization and sterilization by (60)Co, the mean size increased about 10% but no significant change was observed in EY and DL. Tests for in vitro release in physiological saline or physiological saline containing 0.5% (w/v) ascorbic acid by a dialysis bag showed sustained release and little burst effect. PMID- 16316730 TI - Influence of pH, buffer species, and storage temperature on physicochemical stability of a humanized monoclonal antibody LA298. AB - The purpose of this work is to study the effect of pH, buffer species, and temperature on the physicochemical stability of a humanized monoclonal antibody LA298. The study was carried out in solution state of the antibody in the presence of different buffer species at different pH values and storage temperature. No significant changes in total protein content were observed for any of the solutions with different buffers at different pH values when stored for 8 weeks at both 5 degrees C and 25 degrees C or at 37 degrees C for 1 week. Known asparagines (Asn55) deamidation of LA298 was found to be dependent on pH, buffer type, and temperature. The estimated rate constant of the double heavy chain Asn55 deamidation in phosphate buffer at pH 6.5 and 7.0 was much higher than that in citrate buffer under the same storage conditions. However, comparable results were obtained for single heavy chain Asn55 deamidation in citrate and phosphate buffer. Aggregation of LA298 was not significant for samples at different pH values, buffers, and temperatures as the monomer of LA298 decreased dramatically over time. Less decrease in monomeric LA298 was observed in citrate buffer, pH 5.0-5.5. In conclusion, to minimize deamidation and loss of LA298 monomer, it is important to optimize its solution pH, buffer species, and storage temperature. PMID- 16316731 TI - [Morbidity and cost of rotavirus infections in France]. AB - BACKGROUND: Rotavirus is the most common cause of severe diarrhea in children. Morbidity and mortality related to rotavirus infection is not well known in temperate countries in general, and in France in particular. OBJECTIVES: The aim of this study was estimate the morbidity, mortality, and cost related to the rotavirus infection in France, in order to assess the potential impact of a vaccination program. METHODS: A birth cohort was followed until 5 years of age using a decision tree model. Rotavirus infection incidence rates were modeled according to age, seasons, and breast-feeding status. RESULTS: Based on estimates from a decision model, we found that in France, rotavirus infection was responsible for 300,000 annual episodes of acute diarrhea, 138,000 visits to general practitioners, 18,000 hospitalizations, and 9 deaths. The annual direct cost related to rotavirus infection care was estimated at 28 million euros. CONCLUSION: This study demonstrates the high morbidity and cost of care associated with rotavirus infection in France. The decision tree model developed in this study could be used in the future to estimate the potential effectiveness, cost and cost-effectiveness of childhood vaccination strategies using new rotavirus vaccines. PMID- 16316732 TI - MR imaging in probably benign lesions (BI-RADS category 3) of the breast. AB - PURPOSE: To investigate the role of dynamic magnetic resonance (MR) imaging in the evaluation of probably benign lesions (BI-RADS category 3) and its contribution to patient management. MATERIALS AND METHODS: Dynamic breast MR imaging was performed in 56 lesions assessed as probably benign in mammography of 43 patients. In MR imaging, T2-weighted turbo spin echo (TSE) with fat suppression sequence followed by pre- and post-contrast T1-weighted 3D-FLASH sequences were used. MR imaging findings were scored using 0-2 point criterion scale. The lesions were divided into five groups according to their total score (0 point: group 1, negative; 1-2 points: group 2, benign; 3 points: group 3, probably benign; 4-5 points: group 4, suspicious for malignancy; 6-8 points: group 5, highly suggestive of malignancy). Histopathologic verification of lesions in group 4 and above was obtained. Lesions in group 3 were either biopsied or followed up by mammography or MR imaging. Lesions in group 1-2 were followed by mammography of 6-month intervals for 2 years. The sensitivity, specificity, accuracy, and positive and negative predictive values of MR imaging in the determination of malignancy in BI-RADS category 3 lesions were calculated. RESULTS: Fifty-six findings (45 mass, 9 breast tissue, 2 focal enhancement) in 43 patients were detected in MR imaging. According to their total score, 41 lesions (73.2%) and breast tissue had 0 point (group 1); 10 lesions (17.8%) had 1-2 points (group 2); 2 lesions (3.6%) had 3 points (group 3); 2 lesions (3.6%) had 4 and 5 points (group 4); and 1 lesion (1.8%) had 6 points (group 5). Ten lesions (of six in groups 1 and 2, one in group 3, three in groups 4 and 5) were histopathologically confirmed. Out of 10 lesions, only 1 (1.8%) with 4 points in group 4 was diagnosed as invasive ductal carcinoma. Other lesions followed with mammography or MR imaging did not change. The sensitivity, specificity, accuracy, positive and negative predictive values of MR imaging in the determination of malignancy in BI-RADS category 3 lesions were calculated as 100, 96.4, 96.4, 33.3, and 100%, respectively. CONCLUSION: In the evaluation of BI-RADS category 3 lesions, dynamic MR imaging does not provide additional information with low positive predictive value similar to that of short interval mammography follow up. PMID- 16316733 TI - Leucobacter luti sp. nov., and Leucobacter alluvii sp. nov., two new species of the genus Leucobacter isolated under chromium stress. AB - Two strains designated RF6(T) and RB10(T) were isolated, from activated sludge and from river sediments, respectively, both systems receiving chromium contaminated water. Phylogenetic analysis showed that strain RF6(T)and strain RB10(T) represented two new species of the genus Leucobacter. Strain RB10(T) can be distinguished from RF6(T) by its ability to grow at 37 degrees C, by showing a different optimum pH, by cell wall amino acids different relative amount and by having the fatty acid strait C16:0 as the third most abundant fatty acid. On the basis of the distinct peptidoglycan composition, 16S ribosomal DNA sequence analysis, DNA-DNA reassociation values, and phenotypic characteristics we are of the opinion that strain RF6(T) represents a new species of the genus Leucobacter for which we propose the name Leucobacter luti (CIP 108818(T)=LMG 23118) and that strain RB10(T) represents an additional new species of the same genus for which we propose the name Leucobacter alluvii (CIP 108819(T)=LMG 23117). PMID- 16316734 TI - Evaluation of intra-specific diversities in Oenococcus oeni through analysis of genomic and expressed DNA. AB - In winemaking Oenococcus (O.) oeni is the most frequent species of lactic acid bacteria (LAB) associated with malolactic fermentation (MLF). Several studies have demonstrated that O. oeni is a quite homogeneous species and strains are difficult to differentiate especially when isolates from the same region are analyzed. In this study, the molecular biodiversity of O. oeni isolated from wines of the same region (Aglianico produced in Basilicata Region, Southern Italy) was evaluated with the aim of designing a molecular approach for discrimination and characterization of the isolates at the strain level. Three molecular techniques were applied: random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), restriction endonucleases analysis-pulsed field gel electrophoresis (REA-PFGE) and differential display PCR (DD-PCR). The results obtained by RAPD-PCR confirmed the difficulty in differentiating isolates. By means of REA-PFGE a higher polymorphism, often related to the origin (winery) of strains, was revealed. However, on analyzing strains isolated from the same winery, only in some cases was more than one REA-PFGE pattern obtained. By analyzing dendrograms constructed on the basis of DD-PCR profiles differentiation of strains isolated from the same winery, in some cases, could be accomplished. The reliability of the DD-PCR in the differentiation of closely related strains suggests that this method could represent an alternative and/or additional tool to other molecular methods, such as REA-PFGE, for fine characterization of oenococcal strains. PMID- 16316735 TI - Evaluation of recombinant human interferon alpha-2b structure and stability by in gel tryptic digestion, H/D exchange and mass spectrometry. AB - Stability and structure of recombinant interferon alpha-2b (rHuINF alpha-2b) was studied by mass spectrometry (MALDI-TOF and Q-TOF MS), chromatography (LC-UV-FLD DAD, LC-MS) and CD spectroscopy. Besides analysis of the substance according to Ph. Eur. methods, two additional mass spectrometric methods were developed. The aim of both methods was to estimate structure-stability relationship connected to methionine oxidation or protein degradation. Preservation or degradation of protein structure was confirmed by H/D exchange in four separate experiments. Kinetics of deuterium incorporation into macromolecule was monitored over 2670 min. Isoforms of rHuINF alpha-2b were separated by 2D gel electrophoresis. In-gel digestion with trypsin and mass spectrometric analysis, performed on four separated isoforms at the mass corresponding to the mass of rHuINF alpha-2b with oxidized methionines, confirmed oxidation of all methionines to a different extent. Another four isoforms observed in 2D gel are most likely dimers of the same macromolecules with scrambled disulphide bridges. Oxidation and dimerisation are consequences of protein interaction with oxidizing reagents in polyacrilamide gel. PMID- 16316736 TI - Simultaneous determination of nine saponins from Panax notoginseng using HPLC and pressurized liquid extraction. AB - A HPLC and pressurized liquid extraction (PLE) method was developed for simultaneous determination of nine saponins, including notoginsenoside R1, ginsenoside Rg1, Re, Rf, Rb1, Rc, Rb2, Rb3 and Rd in Panax notoginseng. The analysis was performed on C18 column with water-acetonitrile gradient elution and the investigated saponins were authenticated by comparing retention time and mass spectra with their reference compounds. Several methods including PLE, ultrasonication, soxhlet extraction and immersion were used for sample preparation and their extraction efficiency was compared. The results showed that PLE has the highest extraction efficiency and repeatability, which would be valuable on standardization of sample preparation for quality control of Chinese medicines. The developed HPLC and PLE is an effective approach for simultaneously quantitative determination of sapoinins in P. notoginseng, which could be used for quality control of P. notoginseng and its preparations. PMID- 16316737 TI - Structural studies of impurities of risperidone by hyphenated techniques. AB - During the impurity profile of risperidone (RSP), a more polar impurity (RSP-1) and a more non-polar impurity (RSP-2) were detected in LC-MS with respect to risperidone. These impurities were isolated, enriched and were subjected to mass and NMR spectral studies. Based on the spectral data, RSP-1 and RSP-2 were characterized as risperidone N-oxide and 9-methylene risperidone, respectively. The formation of these impurities is rationalized. The structures of both the impurities were unambiguously confirmed by single crystal XRD Studies. PMID- 16316738 TI - Validated LC/MS/MS assay for curcumin and tetrahydrocurcumin in rat plasma and application to pharmacokinetic study of phospholipid complex of curcumin. AB - To study pharmacokinetic properties of curcumin, a fast sensitive assay method was developed to determine curcumin and its metabolite tetrahydrocurcumin in rat plasma. The assay was based on tandem mass spectrometry detection (LC/MS/MS). Salbutamol was used as the internal standard (IS). The method had the lower limit of quantitation (LLOQ) of 0.5 ng/ml in rat plasma, which corresponds to 2.5 pg for the 5 microl injection volume. Good linearity was got to 500 ng/ml. The precision, accuracy, recovery and applicability were found to be adequate for pharmacokinetic studies. Phospholipid complex of the natural compound curcumin was prepared in order to improve its bioavailability. Complex formation resulted in an obvious increase in bioavailability of curcumin in rat in vivo according to the assay by above LC/MS/MS method. PMID- 16316739 TI - Expansion of GAA trinucleotide repeats in mammals. AB - We have previously shown that GAA trinucleotide repeats have undergone significant expansion in the human genome. Here we present the analysis of the length distribution of all 10 nonredundant trinucleotide repeat motifs in 20 complete eukaryotic genomes (6 mammalian, 2 nonmammalian vertebrates, 4 arthropods, 4 fungi, and 1 each of nematode, amoebozoa, alveolate, and plant), which showed that the abundance of large expansions of GAA trinucleotide repeats is specific to mammals. Analysis of human-chimpanzee-gorilla orthologs revealed that loci with large expansions are species-specific and have occurred after divergence from the common ancestor. PCR analysis of human controls revealed large expansions at multiple human (GAA)(30+) loci; nine loci showed expanded alleles containing >65 triplets, analogous to disease-causing expansions in Friedreich ataxia, including two that are in introns of genes of unknown function. The abundance of long GAA trinucleotide repeat tracts in mammalian genomes represents a significant mutation potential and source of interindividual variability. PMID- 16316740 TI - Sequence context analysis in the mouse genome: single nucleotide polymorphisms and CpG island sequences. AB - A genome-wide view of sequence mutability in mice is still limited, although biologists usually assume the same scenario for mice as for humans. In this study, we examined the sequence context in the local environment of 482,528 mouse single nucleotide polymorphisms (SNPs). We found that CpG-containing short sequences, in general, had more representation in the local sequences of SNPs compared to the genome sequences. The extent of this overrepresentation was stronger in mice than in humans, which is inconsistent with previous observations of the weaker neighboring-nucleotide biases on mouse SNPs. To exclude the CpG effect, we compared the distribution patterns of short sequences among the six categories of SNPs. The results revealed an even stronger pattern in the CpG containing group for C/G substitution compared to for A/G or C/T substitutions. We next performed the first genome-wide sequence context analysis of SNPs in the mouse CpG islands. SNPs occurring at CpG sites were 3.14-fold less prevalent than expected, suggesting the suppression of methylation-dependent deamination in the CpG islands. The extent of this suppression was less in mice than in humans. Finally, compared with humans, the observations of a greater deficit of CpG dinucleotides, a stronger overrepresentation of CpG-containing n-mers surrounding the polymorphic sites, and a higher SNP/genome ratio of CpG dinucleotides in the mouse genome support the "loss of CpG islands" model in the mouse lineage. PMID- 16316741 TI - Uterine rupture after 400 micrograms misoprostol oral administration in the first trimester of pregnancy. PMID- 16316743 TI - Effect of neonatal isolation on outcome following neonatal seizures in rats--the role of corticosterone. AB - Emerging evidence indicates that early maternal care permanently modifies the activity of hypothalamic-pituitary-adrenal (HPA) axis and is a critical factor in determining the capacity of the brain to compensate for later encountered insults. The purpose of this study was to determine the role of corticosterone (CORT) in the detrimental effects of neonatal isolation (NI) on seizures. Rats were assigned randomly to the following five groups: (1) control (CONT) rats; (2) NI rats that underwent daily separation from their dams from postnatal day 2 (P2) to P9; (3) status epilepticus (SE) rats, induced by lithium-pilocarpine (Li-Pilo) model at P10; (4) NI plus SE (NIS) rats and (5) NISM rats, a subset of NIS rats receiving metyrapone (100 mg/kg), a CORT synthesis inhibitor, immediately after SE induction. At P10, plasma CORT levels were compared at baseline in CONT and NI rats and in response to Li-Pilo-induced SE among SE, NIS and NISM rats. We evaluated the spatial memory in the Morris water maze at P50 approximately 55, the expression of hippocampal cyclic adenosine monophosphate (cAMP)-responsive element-binding protein phosphorylation at serine-133 (pCREBSer-133) at P55, hippocampal neuronal damage at P80 and seizure threshold at P100. The isolated rats exhibited higher CORT release in response to SE than non-isolated rats, and the NIS rats had greater cognitive deficits and decreased seizure threshold compared to the CONT, NI and SE groups. By contrast, the NISM group, compared to the NIS group, showed a normal CORT response to SE and better spatial memory but no difference in seizure threshold. Compared to the CONT group, the hippocampal pCREBSer-133 level was significantly reduced in all experimental groups (NI, SE, NIS, NISM) with no differences between groups. All rats were free of spontaneous seizures later in life and had no discernible neuronal loss in the hippocampus. Results in this model demonstrate repetitive NI enhances response of plasma CORT to SE, and exacerbates the neurological consequences of neonatal SE. Amelioration of neurological sequelae following reduction of the SE-induced excessive rise in plasma CORT implicates CORT in the pathogenesis of NI increasing the vulnerability to seizures. PMID- 16316744 TI - Zonisamide as adjunctive therapy for refractory partial seizures. AB - Zonisamide (Zonegran) has been used extensively worldwide (>2 million patient years experience) for the effective treatment of a broad range of epilepsy indications. Four randomised, placebo-controlled trials (duration or=300 mg/day to be efficacious in treating refractory partial seizures in adults. In a pivotal European study, zonisamide 500 mg/day was significantly superior to placebo in reducing the frequency of complex partial seizures (-51% versus -16%), all partial seizures and all seizures, with dose dependent benefit provided over a 100-500 mg/day dose range. Supporting trials have confirmed significant increases in reduction in median seizure frequency (up to 41%) and responder rates (35-42%) compared with placebo following zonisamide 400-600 mg/day, enabling 20-27% of patients to attain >or=75% reduction in seizure frequency. Pooled data from all four placebo-controlled trials demonstrate an excellent tolerability and safety profile; adverse events are generally of mild-moderate severity with few leading to discontinuation, and incidence of serious adverse events is comparable to placebo. These data support the use of zonisamide in combination with commonly used antiepileptic drugs to provide efficacious and well-tolerated treatment for patients with refractory partial seizures. PMID- 16316745 TI - Antimicrobial potentiality of the thioxanthene flupenthixol through extensive in vitro and in vivo experiments. AB - The antipsychotic thioxanthene flupenthixol, possessing a trifluoromethyl substituent at position 2, exhibited a distinct antibacterial property against 352 strains of bacteria from 3 Gram-positive and 13 Gram-negative genera. The minimum inhibitory concentration (MIC) of flupenthixol was determined by the National Committee for Clinical Laboratory Standards agar dilution method. MICs ranged from 10-100 microg/mL in most of the strains, whilst some strains were inhibited at even lower concentrations. The mode of action of this drug was found to be bacteriostatic against Staphylococcus aureus and Vibrio cholerae. In the in vivo experiments, this drug was capable of contributing significant protection (P < 0.001) to a Swiss strain of white mice challenged with 50 median lethal dose of a mouse-virulent strain at a drug concentration of 15 microg/mouse. In addition, flupenthixol remarkably reduced the number of viable bacteria in organ homogenates and blood of mice treated with this drug. PMID- 16316746 TI - A double-blind, randomised, controlled clinical trial of acetyl-L-carnitine vs. amisulpride in the treatment of dysthymia. AB - AIM: Evaluation of the effect of acetyl-L-carnitine (ALCAR) vs. amisulpride measured by total Hamilton Depression Rating Scale score (HAM-D(21)) in patients with pure dysthymia (DSM IV). Two hundred and four patients were randomised and treated with ALCAR 500 mg b.i.d. or amisulpride 50 mg u.i.d. in a double-blind study, for 12 weeks. RESULTS: A solid improvement of HAM-D(21) was observed in both treatment groups throughout the study. The results did not disclose statistically significant differences between treatments, although the confidence interval for the non-inferiority of the primary end-point exceeded the pre established limit of 2 by 0.46 points. According to a non-inferiority margin of 3 (considered acceptable by recent published data) the primary end-point could have been fully satisfied. CDRS, MADRS and CGI, employed to further measure the clinical outcome, reported similar results in both treatment groups. The greater tolerability of ALCAR is of clinical relevance considering the chronicity of dysthymia, which often requires prolonged treatment. PMID- 16316747 TI - On the problem of field-gradient NMR measurements of intracrystalline diffusion in small crystallites--water in NaA zeolites as an example. AB - Necessary conditions for measuring intracrystalline diffusion in small crystal size systems via field-gradient NMR are discussed. As an illustrative case self diffusion coefficients of water adsorbed in NaA zeolites (average crystal diameter about 1 microm) have been measured by 1H-NMR stimulated echoes in static magnetic field gradients of up to 180 T/m in the temperature range of 254-344 K. Obtaining intracrystalline diffusion coefficients necessitates a sufficiently high spatial resolution only provided by such large field gradients. PMID- 16316748 TI - [Nephrotic syndrome in Congolese adolescents: clinical and pathological findings]. PMID- 16316749 TI - In vitro uptake of gelatin nanoparticles by murine dendritic cells and their intracellular localisation. AB - The long term goal of this study is to develop an efficient nanoscopic vaccine delivery system, based on the biodegradable and natural polymer gelatin, to deliver therapeutic protein antigens along with adjuvants into dendritic cells (DCs). In this study, gelatin nanoparticles were tested for qualitative and quantitative uptake in murine DCs in vitro. A second aim of this study was to prove that the carrier system is able to deliver tetramethylrhodamine conjugated dextran (TMR-dextran), as a model drug into the DCs. The TMR-dextran was incorporated during the preparation of the gelatin nanoparticles. DCs were generated from murine bone marrow cells by an established ex vivo technique. Flow cytometry showed that 88% of the cells positive for the specific murine DC marker CD11c took up TMR-dextran loaded gelatin nanoparticles, whereas only 4% of the soluble form of TMR-dextran was taken up. Double color confocal laser scanning microscopy (CLSM) showed that gelatin nanoparticles were phagocytosed by DCs and the triple color CLSM showed that the TMR-dextran was localized mainly in lysosomes as expected, but partly also outside the lysosomes, presumably in the cytoplasm. An in vitro release study of TMR-dextran from gelatin nanoparticles demonstrated that there was hardly any release in phosphate buffered saline (PBS), but by trypsin-assisted degradation of gelatin nanoparticles resulted in the release of about 80% of the TMR-dextran from the particles. These results suggest that gelatin nanoparticles hold promise as a new biocompatible tool for vaccine delivery to DCs, with applications in cancer immunotherapy. PMID- 16316750 TI - The wide diversity and complexity of peptides bound to class II MHC molecules. AB - The identification and quantitation of peptides selected by class II MHC molecules during natural processing of proteins is of key importance in understanding the repertoire and distribution of T cells. The examination of peptides selected by class II MHC molecules has depended greatly on mass spectrometry, a powerful technique that identifies and sequences peptides in complex mixtures with great sensitivity and precision. Such analysis has resulted in the identification of several factors, including the repertoire of peptides selected by MHC molecules during natural processing of proteins, motifs important for selection of processed peptides, conformational isomers of peptide-MHC complexes, and post-translational changes to the peptides. PMID- 16316751 TI - Cdc48p is UBX-linked to ER ubiquitin ligases. AB - Proteasome-mediated turnover of misfolded secretory and transmembrane proteins at the cytoplasmic face of the endoplasmic reticulum (ER) membrane is dependent on a AAA-ATPase complex formed by the ubiquitin-selective chaperone Cdc48p in Saccharomyces cerevisiae and mammals by the Cdc48p homologue p97. Two new papers reveal that the Ubx2 protein physically links ER-membrane-integrated ubiquitin ligases to Cdc48p, and that it is essential for degradation of substrates that are ubiquitylated at the cytoplasmic face of the ER. PMID- 16316752 TI - A response to issues raised in a recent paper concerning the Oxford knee score. AB - In a recently published paper, authors were critical of the Oxford Knee Score (OKS) patient based measure. We discuss a number of the interesting issues that this paper raised and point out some obvious misunderstandings. The OKS, whilst not perfect, has been shown in independent comparative studies to perform more satisfactorily than other measures. It remains one of a small number of measures with satisfactory measurement properties. It would be a great pity if clinicians were deterred from using the OKS on the basis of comments made in the recent publication. PMID- 16316753 TI - Soybean mosaic virus helper component-protease enhances somatic embryo production and stabilizes transgene expression in soybean. AB - Soybean mosaic virus (SMV) helper component protease (HC-Pro), a suppressor of post-transcriptional gene silencing, was evaluated for its ability to enhance production of soybean hygromycin-resistant somatic embryos (HR-SEs), and stabilize transgene expression. Immature soybean cotyledonary explants were co cultured with Agrobacterium tumefaciens strain KYRT1 harboring either pCAMBIA1302, carrying a hygromycin phosphotransferase gene (hpt) and a gene encoding green fluorescent protein; pCAMBIA1305.1, carrying hpt and beta glucuronidase (uidA) genes; pG2-HC-Pro, a derivative of pCAMBIA1305.1 containing SMV G2 HC-Pro; or pG5-HC-Pro, a derivative of pCAMBIA1305.1 containing SMV G5 HC Pro, but lacking uidA. Significantly (rho<0.02) higher numbers of HR-SEs were obtained from explants transformed with Agrobacterium harboring either pG2-HC-Pro or pG5-HC-Pro than with either of the vector controls (pCAMBIA1302 or pCAMBIA1305.1). Beta-glucuronidase (GUS) expression was significantly (rho<0.003) higher in 50-day-old transgenic plants expressing GUS along with SMV-HC-Pro and in SMV-infected GUS transgenic plants than in transgenic plants expressing GUS alone. Together, these data suggest that SMV-HC-Pro enhanced recovery of HR-SEs by suppressing silencing of the hygromycin phosphotransferase gene. PMID- 16316754 TI - Budded karyoplasts from multinucleated fibroblast cells contain centrosomes and change their morphology to mitotic cells. AB - Cell changes to accelerated rates of growth are determined by random, new gene mutations that have their origin in vitro in a morphologically visible process of cell alterations. It is a continuous process that, step by step, transforms the normal cell into extended life (EL) cells. These latter cells with limited life spans can further transform to immortalized cells (i.e., cell lines) by the same sequence of morphological cell changes. In contrast to human epithelial cells, fibroblasts in culture have not given rise spontaneously to EL cells. Therefore, it was assumed that some of the cell changes (i.e., cell indicators of the process of transformation) might not be present in near senescent fibroblast cell cultures. As a positive control to normal fibroblast expansion to senescence, the same cells were stressed by inadequate nutrition and confluence. Another positive control was cell indicators induced by SV40 infections. The consecutive sequence of the cell indicators in the transformation process reported previously were: (1) large polyploid cells with nuclei that contained more than two genomes, (2) fragmentation/amitosis of the polyploid nuclei to bi- and multinucleated cells (MNCs), and (3) nuclear budding (i.e., karyoplasts) from MNCs that gave rise to EL cell colonies with various longevities beyond the senescent phase. The present study shows that MNCs and karyoplasts were present in the near-senescent fibroblast cell cultures. Furthermore, new data on the following aspect of the cell transformation process are presented: (1) association of the nuclear fragmentation process with death of cells, (2) cytological markers that distinguish between fragmentation-MNCs versus MNCs from cell fusion, (3) cytological changes of karyoplasts that go through mitotic division to produce daughter cells, and (4) presence of two centrosomes (spindle polar bodies) in the budded karyoplasts. These new findings are discussed in regard to the nuclear fragmentation process in polyploid cells which gives rise to smaller, viable nuclei in MNCs with reduced numbers of whole genomes. PMID- 16316755 TI - Cell reproduction and genome multiplication in the proliferative and invasive trophoblast cell populations of mammalian placenta. AB - Spatiotemporal "time-table" of ways of cell reproduction (mitosis, restitutional mitosis, endomitosis, endoreduplication) of trophoblast cell populations is described. The populations of mitotically active trophoblast cells (diploid and low-polyploid) are located mostly out of contact with maternal tissues. In rodent placenta they mainly switch from mitotic cycle to polyploidizing (restitutional) mitoses and reach 4c-8c. Thereafter they switch to endoreduplication and reach 16c-64c. Following a series of endoreduplication cycles a part of this cell population sets apart and penetrates deeply into the decidualized endometrium and myometrium, their capabilities for replication being lost progressively (in rodent--256c-1024c). The invasive trophoblast cells that reach 256c-1024c via endoreduplication simultaneously form a barrier between semiallogenic fetal and maternal tissues. Arrest of mitoses and complete repression of DNA replication after a series of endoreduplication cycles makes hardly probable the renewal of mitotic activity in the deeply invading tertiary giant trophoblast cells, thereby preventing the possibility of their ectopic expanding in the maternal tissues during the normal pregnancy. PMID- 16316756 TI - Neosis--a paradigm of self-renewal in cancer. AB - We recently described a novel form of cell division termed neosis, which appears to be the mode of escape of cells from senescence and is involved in the neoplastic transformation and progression of tumors (Cancer Biol & Therap 2004;3:207-18). Neosis is a parasexual somatic reduction division and is characterized by (1) DNA damage-induced senescence/mitotic crisis and polyploidization, (2) followed by production of aneuploid daughter cells via nuclear budding, (3) asymmetric cytokinesis and cellularization conferring extended, but, limited mitotic life span to the offspring, and (4) is repeated several times during tumor growth. The immediate neotic progeny are termed the Raju cells, which seem to transiently display stem cell properties. The Raju cells immediately undergo symmetric mitotic division and become mature tumor cells. Exposure of tumor cells to genotoxic agents yields neosis-derived Raju cell progenies that are resistant to genotoxins, thus contributing to the recurrence of drug-resistant tumor growth. Similar events have been described in the literature under different names through several decades, but have been neglected due to the lack of appreciation of the significance of this process in cancer biology. Here we review and interpret the literature in the light of our observations and the recent advances in self-renewal in cancer. Neosis paradigm of self-renewal of cancer growth is consistent with the telomere attrition, aging and origin of cancer cells after reactivation of telomerase, and constitutes an alternative to the cancer stem cell hypothesis. We summarize the arguments favoring Raju cells and not cancer stem cells, as the source of self-renewal in cancer and present a comprehensive hypothesis of carcinogenesis, encompassing various aspects of cancer biology including senescence, tumor suppressor genes, oncogenes, cell cycle checkpoints, genomic instability, polyploidy and aneuploidy, natural selection, apoptosis, endoapoptosis, development of resistance to radiotherapy and chemotherapy leading tumor progression into malignancy. PMID- 16316757 TI - Getting the right blood to the right patient: the contribution of near-miss event reporting and barrier analysis. AB - Safety and reliability in blood transfusion are not static, but are dynamic non events. Since performance deviations continually occur in complex systems, their detection and correction must be accomplished over and over again. Non conformance must be detected early enough to allow for recovery or mitigation. Near-miss events afford early detection of possible system weaknesses and provide an early chance at correction. National event reporting systems, both voluntary and involuntary, have begun to include near-miss reporting in their classification schemes, raising awareness for their detection. MERS-TM is a voluntary safety reporting initiative in transfusion. Currently 22 hospitals submit reports anonymously to a central database which supports analysis of a hospital's own data and that of an aggregate database. The system encourages reporting of near-miss events, where the patient is protected from receiving an unsuitable or incorrect blood component due to a planned or unplanned recovery step. MERS-TM data suggest approximately 90% of events are near-misses, with 10% caught after issue but before transfusion. Near-miss reporting may increase total reports ten-fold. The ratio of near-misses to events with harm is 339:1, consistent with other industries' ratio of 300:1, which has been proposed as a measure of reporting in event reporting systems. Use of a risk matrix and an event's relation to protective barriers allow prioritization of these events. Near-misses recovered by planned barriers occur ten times more frequently then unplanned recoveries. A bedside check of the patient's identity with that on the blood component is an essential, final barrier. How the typical two person check is performed, is critical. Even properly done, this check is ineffective against sampling and testing errors. Blood testing at bedside just prior to transfusion minimizes the risk of such upstream events. However, even with simple and well designed devices, training may be a critical issue. Sample errors account for more than half of reported events. The most dangerous miscollection is a blood sample passing acceptance with no previous patient results for comparison. Bar code labels or collection of a second sample may counter this upstream vulnerability. Further upstream barriers have been proposed to counter the precariousness of urgent blood sample collection in a changing unstable situation. One, a linking device, allows safer labeling of tubes away from the bedside, the second, a forcing function, prevents omission of critical patient identification steps. Errors in the blood bank itself account for 15% of errors with a high potential severity. In one such event, a component incorrectly issued, but safely detected prior to transfusion, focused attention on multitasking's contribution to laboratory error. In sum, use of near-miss information, by enhancing barriers supporting error prevention and mitigation, increases our capacity to get the right blood to the right patient. PMID- 16316758 TI - The reality of self-sufficiency. AB - In this article, some points concerning the self-sufficiency in Europe will be discussed. After the definition of the self-sufficiency, the situation in Central and Eastern Europe will be briefly presented, as well as the problems connected with national and community self-sufficiency. PMID- 16316759 TI - [The impact of 18F-fluorodeoxyglucose positron emission tomography on the 3D conformal radiotherapy planning in patients with non-small cell lung cancer]. AB - Traditional radiation treatment planning relies on density imaging such as Computed Tomography for anatomic information of various structures of interest including target and normal tissues. However, the difficulties to distinguish malignant from normal tissue on CT slides often leads to inaccurate outlining of the GTV and/or to geographic misses. 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) has shown an increase in both sensitivity and specificity over CT in locoregional staging of patients with non-small cell lung cancer (NSCLC). The co registration of FDG-PET images to the data of the CT planning offers the radiation oncologist the possibility to include functional information into the target outlining. For the treatment of patients with NSCLC, it has been shown that the use of FDG-PET images: 1) modified the shape and volume of radiation fields in 22-62% of cases, mainly due to a better nodal staging and distinction of atelectasis from tumor and; 2) significatively reduced the interobserver and intraobserver variability. This paper reviews the results reported in the literature. Challenges and proposed solutions are discussed. PMID- 16316760 TI - [Uterine arteriovenous malformation. A rare cause of recurrent metrorrhagia]. PMID- 16316761 TI - [The consensus statement "Pregnancy and tobacco" (Lille, 7-8 October, 2004)]. PMID- 16316763 TI - [Assisted reproductive technologies after conservative management of borderline or invasive ovarian tumours]. PMID- 16316764 TI - [Tocolysis with nifedipine: its use in current practice]. PMID- 16316765 TI - [A prospective study of the provision of emergency contraception in family planning centers in Val-de-Marne]. PMID- 16316766 TI - [Tocolysis with nifedipine: its use in current practice]. PMID- 16316767 TI - [A prospective study of the provision of emergency contraception in family planning centers in Val-de-Marne]. PMID- 16316768 TI - [Laparoscopic treatment of borderline ovarian tumor: analysis of 54 patients and clinical outcomes]. PMID- 16316769 TI - [Biochemical markers of bone remodeling]. AB - Bone markers are aimed to assess bone cells activity. Some are specific of bone formation (osteocalcin, bone alkaline phosphatase, extension peptides of type I procollagen); others are specific of bone resorption (deoxypyridinoline and peptidebound forms.). Their main applications are: evaluation of fracture risk and follow-up of antiosteoporotic treatments. Their use in clinical practice is difficult: choice of the most appropriate bone marker, relevance of changes, and may require the specialist's opinion. PMID- 16316770 TI - [GnRH analogues and myomas: which strategy?]. PMID- 16316771 TI - [Prognosis work-up in prenatal medicine: the example of Down's syndrome]. PMID- 16316772 TI - [Ovarian endometrioma and infertility: when not to treat?]. PMID- 16316773 TI - [GnRH analogues and myomas: somewhat for]. PMID- 16316774 TI - Long-term treatment outcome of oral premalignant lesions. AB - The purpose of the present retrospective study was to learn the long-term outcome of oral premalignant lesions, leukoplakia and erythroplakia, with or without surgical intervention and to relate the outcome to factors supposed to be significant for malignant development including clinical type, demarcation, size, site, presence of epithelial dysplasia, smoking and surgery. A total of 269 lesions in 236 patients were included. Ninety-four lesions were surgically removed, 39 lesions (41%) being homogenous and 46 (49%) non-homogenous leukoplakias whereas nine (5%) were erythroplakias. Seventy-three percent of the lesions were associated with tobacco habits. The mean size of the lesions was 486 mm(2), and 71% of the lesions showed a degree of epithelial dysplasia. After excision the defects were closed primarily by transposition of mucosal flaps or they were covered by free mucosal or skin grafts. A few defects were left for secondary healing. After surgical treatment the patients were followed (mean 6.8 yrs, range 1.5-18.6 yrs), and new biopsies taken in case of recurrences. One hundred and seventy five lesions had no surgical intervention, 149 lesions (85%) being homogenous and 20 (11%) non-homogenous leukoplakias, and 6 (3%) erythroplakias. Eighty-one percent of the lesions were associated with smoking. The mean size of the lesions was 503 mm(2) and 21 of the lesions (12%) exhibited epithelial dysplasia. Sixty-five lesions were not biopsied. These patients were also followed (mean 5.5 yrs, range 1.1-20.2 yrs), and biopsies taken in case of changes indicative of malignant development. All patients were encouraged to quit smoking and candidal infections were treated. The possible role of different variables for malignant development was estimated by means of logistic regression analysis. Following surgical treatment 11 lesions (12%) developed carcinoma after a mean follow-up period of 7.5 yrs. Non-homogenous leukoplakia accounted for the highest frequency of malignant development, i.e. 20%, whereas 3% of the homogenous leukoplakias developed carcinomas. Surgically treated lesions with slight, moderate, severe and no epithelial dysplasia developed carcinoma with similar frequencies, i.e. 9-11%. Without surgical intervention 16% of the 175 lesions disappeared whereas seven lesions (4%) developed carcinoma after a mean observation period of 6.6 yrs. The highest frequency of malignant development (15%) was seen for non-homogenous leukoplakias, this figure being 3% for homogenous leukoplakias. Fourteen percent of lesions with slight epithelial dysplasia developed malignancy and 2% of lesions with no dysplasia showed malignant transformation. Logistic regression analysis showed a seven times increased risk (OR = 7.0) of non-homogenous leukoplakia for malignant development as compared with homogenous leukoplakia and a 5.4 times increased risk for malignant development for lesions with a size exceeding 200 mm(2). No other examined variables including presence of any degree of epithelial dysplasia, site, demarcation, smoking and surgical intervention were statistically significant factors for malignant development. PMID- 16316775 TI - Clinical and prognostic value of [(18)F]FDG-PET for surveillance of oral squamous cell carcinoma after surgical salvage therapy. AB - [(18)F]FDG-PET was found to be useful for recurrence detection in patients with oral squamous cell carcinoma (OSCC), as a negative PET scan predicted a favorable outcome and survival. Here, we evaluate PET performance in the management of OSCC patients with recurrent/second primary disease after potentially curative second line therapy. Forty one OSCC patients underwent salvage surgery and 31/41 had received radiation therapy. Thirty five/41 developed recurrent and 6/41 second primary OSCC. Patients had PET evaluation 8.4months (median) after surgery and were followed for at least 6months until disease recurrence or death. For surviving patients, the median follow-up was 33.6months after PET. In OSCC patients who had undergone potentially curative second-line therapy, PET had an overall sensitivity of 85% (92% for recurrence or second primaries, 88% for lymph node failure and 73% for distant metastases). Overall survival was 71% in the PET negative group and 35% in the PET positive group (p<0.01, log-rank test). Moderate glucose metabolism (standardized uptake value4) suggested promising outcome, while SUV>4 indicated a fatal disease course. The data suggest that [(18)F]FDG-PET can facilitate re-staging and clinical management in "high-risk" patients with OSCC. PMID- 16316776 TI - Leukocyte activation by malarial pigment. AB - Malarial pigment, a unique hemozoin crystal composed of unit cells of heme dimers, is present in large amounts in circulating monocytes and neutrophils and can persist unchanged in macrophages for several months. In the present study, we investigated the effect of hemozoin not only on macrophages, but also on neutrophils. We used beta-hematin (BH), a chemically synthetic crystal structurally identical to hemozoin, for these studies. In vitro, BH up-regulated the expression of tumor necrosis factor-alpha in whole blood and in isolated peritoneal macrophages, indicating that hemozoin is able to stimulate monocytes. BH stimulated murine peritoneal neutrophils to express macrophage inflammatory protein-2 (MIP-2), a homologue of human interleukin-8 that is used as a marker of neutrophil activation. Injecting BH into the peritoneal cavity resulted in a dose dependent migration of neutrophils and a high level of myeloperoxidase activity of peritoneal cells. Finally, BH directly induced neutrophil chemotaxis in vitro. Taken together, these results suggest that the malarial pigment hemozoin can activate leukocytes and may participate in the pathology of severe malaria. PMID- 16316777 TI - Comparison of oral fluid and serum ELISAs in the determination of IgG response to natural human papillomavirus infection in university women. AB - BACKGROUND: Venipuncture (phlebotomy) is an obstacle to subject recruitment and ongoing participation in cohort studies investigating human papillomavirus (HPV) infection. Anti-HPV antibodies are not only detected in serum but also in oral fluid. OBJECTIVES: To evaluate if oral fluid specimens can be used in lieu of blood specimens for determining HPV antibody status. STUDY DESIGN: One hundred and seven paired oral fluid and blood specimens from female university students were tested in a HPV16 ELISA and compared to sexual history and serial genital HPV16 DNA status. RESULTS: ELISA results were in agreement in 97% (104/107) of paired sera and oral fluid. Of six women with positive anti-HPV16 serum samples, only three had positive oral fluid specimens. However, the specificity of the oral fluid test was 100% compared to the blood test. CONCLUSIONS: Detection of antibodies in oral fluid correlated with antibodies but was less sensitive than sera. A larger validation study is required to fully characterize the oral fluid assay. PMID- 16316778 TI - Adefovir added to lamivudine for hepatitis B recurrent infection in refractory B cell chronic lymphocytic leukemia on prolonged therapy with Campath-1H. AB - We describe a case of severe reactivation of occult hepatitis B virus infection in a 49-year-old man, who was treated with high doses of chlorambucil for a Binet stage A B-cell chronic lymphocytic leukemia (B-CLL). The patient was initially treated with lamivudine and subsequently with lamivudine and adefovir dipivoxil combination therapy to control viral replication and allow for long-term anti cancer chemotherapy with alemtuzumab (Campath-1H), which was introduced to rescue for a B-CLL relapse. During 20 months of anti-HBV therapy, ALT and HBV-DNA levels progressively declined and B-CLL was successfully kept under control by long-term alemtuzumab administration. PMID- 16316779 TI - Hepatitis C virus superinfection in hepatitis B virus chronic carriers: a reciprocal viral interaction and a variable clinical course. AB - BACKGROUND: The virological and clinical impact of hepatitis C virus (HCV) superinfection in chronic hepatitis B virus (HBV) carriers has been poorly characterized. OBJECTIVE: To evaluate the viral interaction, clinical presentation and course of the disease in four HBsAg/HBV-DNA positive chronic hepatitis patients who developed acute HCV infection. STUDY DESIGN: To evaluate clinical, virological and laboratory data for at least 6 months from the onset of acute HCV infection in patients with chronic HBV infection. RESULTS: Three patients with acute HCV infection had a normal clinical course, but the remaining patient had severe disease with ascites and a marked decrease in prothrombin activity. In all cases, plasma HBV-DNA, which had been detectable prior to the HCV infection, was no longer detectable when the acute HCV infection occurred. The inhibition exerted by HCV on HBV-DNA persisted throughout the follow-up period in three patients, but was temporary in the one patient who experienced an acute exacerbation of chronic HBV infection. HCV-RNA became persistently undetectable in two patients and reduced to low levels in the other two. CONCLUSIONS: Acute HCV infection in the four HBV chronic carriers was characterized by a reciprocal inhibition of HBV-HCV genomes and, in one case, by a severe course of disease. PMID- 16316780 TI - Development of the 40Hz steady state auditory evoked magnetic field from ages 5 to 52. AB - OBJECTIVE: Adults exhibit strong auditory 40 Hz magnetic steady state responses (SSR). Although EEG measured SSR has been studied in children, the developmental course of the magnetic SSR is unknown. METHODS: Sixty-nine healthy subjects ranging in age from 5 to 52 years participated in a magnetoencephalographic (MEG) study. Stimuli were monaural 500 ms duration click trains with a 25 ms inter click interval. Contralateral magnetic responses for both hemispheres were recorded with a 37-channel MEG system. Responses were averaged and examined using wavelet-based time-frequency analysis. Source analyses were also conducted on a subset of the data. RESULTS: Gamma power from 200 to 500 ms post-stimulus onset was computed and was significantly related to subject age in both hemispheres. Hemispheric asymmetry was observed for the anterior-posterior SSR source locations, suggestive of asymmetry similar to that previously described for the SSR and other auditory evoked magnetic field components. CONCLUSIONS: The 40 Hz power findings are generally consistent with previous EEG studies of steady state responses in children showing age-related changes in the 40 Hz SSR. SIGNIFICANCE: Age-related changes in the strength of the magnetic 40 Hz SSR may continue to develop well beyond early childhood, which should be taken into consideration in planning future studies using adolescents and young adults. PMID- 16316781 TI - Alpha and beta changes in cortical oscillatory activity in a go/no go randomly delayed-response choice reaction time paradigm. AB - OBJECTIVE: Predictable movements induce oscillatory changes over the contralateral motor cortex that begin before the movement, but their significance has not been fully established. We studied non-phase-locked changes in cortical oscillatory activity in a S1-centred double-stimulus go/no go paradigm with random interstimulus interval. METHODS: About 58 reference-free EEG channels were analyzed by means of Gabor transforms in a group of 10 healthy subjects. A 2000 Hz tone (S1go, 84% probability) indicated the subject to make a brisk wrist extension after a second 2000 Hz tone (S2go). The S1-S2 interval was either 1.5, 3 or 4.5 s. A 1000 Hz tone (S1 no go, 16% probability) indicated the subject not to move (and wait for another S1 tone). RESULTS: A frontal 15 Hz synchronization was observed after S1 in all conditions. No further significant changes were observed in the no go condition. A small pre-S2 alpha and beta desynchronization could be observed only in the 3 and 4.5s-interval go conditions, being larger in the latter. CONCLUSIONS: These results suggest that the predictability of the timing of a movement influences the appearance of the pre-movement oscillatory changes; not only motor planning (the 'go' decision) is necessary, but also an estimation of when to move. SIGNIFICANCE: Our findings provide new insight on the relationship between the decision-making process, movement, and cortical oscillatory activity. PMID- 16316782 TI - Aging changes and gender differences in response to median nerve stimulation measured with MEG. AB - OBJECTIVE: The current study uses magnetoencephalography (MEG) to characterize age-related changes and gender differences in the amplitudes and timing of cortical sources evoked by median nerve stimulation. METHODS: Thirty-four healthy subjects from two age groups: 20-29 and >64 years of age were examined. After measuring the MEG responses, we modeled the data using a spatio-temporal multi dipole modeling approach to determine the source locations and their associated timecourses. RESULTS: We found early, large amplitude responses in the elderly in primary somatosensory (approximately 20 ms) and pre-central sulcus timecourses (approximately 22 ms) and lower amplitude responses in the elderly later in primary somatosensory (approximately 32 ms) and contralateral secondary somatosensory timecourses (approximately 90 ms). In addition, females had larger peak amplitude responses than males in the contralateral secondary somatosensory timecourse (approximately 28 and 51 ms). CONCLUSIONS: These results show that the median nerve stimulation paradigm provides considerable sensitivity to age- and gender-related differences. The results are consistent with the theory that increased amplitudes identified in the elderly may be associated with decreased inhibition. SIGNIFICANCE: The results emphasize that an examination of two discrete age groups, collapsed across gender, cannot provide a complete understanding of the fundamental changes that occur in the brain across the lifetime. PMID- 16316784 TI - Glucocorticosteroid therapy decreases CD14-expression and CD14-mediated LPS binding and activation of monocytes in patients suffering from systemic lupus erythematosus. AB - In order to study the possible action of glucocorticosteroids (GCS) on the CD14/Toll like receptor mediated activation of monocytes the CD14-expression, CD14-mediated LPS binding and activation of these cells of patients suffering from Systemic Lupus Erythematosus receiving no, low dose or pulse steroid treatment was studied. The CD14-expression was determined on whole blood monocytes by flow cytometry, while the LPS-binding of an FITC-LPS preparate and the LPS-induced TNFalpha secretion were tested on isolated monocytes. The CD14 dependent and -independent LPS-binding and activation were evaluated with the help of a blocking anti-CD14 mAb. Our results showed that the CD14-expression, CD14-dependent LPS-binding and activation were significantly inhibited by the in vivo applied pulse steroid therapy. In contrast, the CD14-independent LPS-binding and activation were not altered by the GCS treatment. Our data provide further in vivo evidence for a possible new way of GCS therapy is able to initiate its anti inflammatory action. PMID- 16316783 TI - Cytokines sing the blues: inflammation and the pathogenesis of depression. AB - Increasing amounts of data suggest that inflammatory responses have an important role in the pathophysiology of depression. Depressed patients have been found to have higher levels of proinflammatory cytokines, acute phase proteins, chemokines and cellular adhesion molecules. In addition, therapeutic administration of the cytokine interferon-alpha leads to depression in up to 50% of patients. Moreover, proinflammatory cytokines have been found to interact with many of the pathophysiological domains that characterize depression, including neurotransmitter metabolism, neuroendocrine function, synaptic plasticity and behavior. Stress, which can precipitate depression, can also promote inflammatory responses through effects on sympathetic and parasympathetic nervous system pathways. Finally, depression might be a behavioral byproduct of early adaptive advantages conferred by genes that promote inflammation. These findings suggest that targeting proinflammatory cytokines and their signaling pathways might represent a novel strategy to treat depression. PMID- 16316785 TI - Histopathological and behavioral improvement of murine mucopolysaccharidosis type VII by intracerebral transplantation of neural stem cells. AB - The therapeutic efficacy of neural stem cell transplantation for central nervous system (CNS) lesions in lysosomal storage disorders was explored using a murine model of mucopolysaccharidosis type VII (MPS VII). We used fetal neural stem cells derived from embryonic mouse striata and expanded in vitro by neurosphere formation as the source of graft materials. We transplanted neurospheres into the lateral ventricles of newborn MPS VII mice and found that donor cells migrated far beyond the site of injection within 24 h, and some of them could reach the olfactory bulb. A quantitative measurement indicated that the GUSB activity in the brain was 12.5 to 42.3% and 5.5 to 6.3% of normal activity at 24 h and 3 weeks after transplantation. In addition, histological analysis revealed a widespread decrease in lysosomal storage in the recipient's hippocampus, cortex, and ependyma. A functional assessment with novel-object recognition tests confirmed improvements in behavioral patterns. These results suggest that intracerebral transplantation of neural stem cells is feasible for treatment of CNS lesions associated with lysosomal storage disorders. PMID- 16316786 TI - Nitric oxide production by hemocytes of larva and pharate prepupa of Galleria mellonella in response to bacterial lipopolysaccharide: Cytoprotective or cytotoxic? AB - Nitric oxide production by the hemocytes of the last instar larvae and sessile pharate prepupa of Galleria mellonella (Lepidoptera: Pyralidae) was demonstrated in vitro in response to preparations of bacterial lipopolysaccharide (LPS) from Escherichia coli using the Griess reaction. Augmented, dose dependent nitric oxide production was observed in the pharate prepupal hemocytes compared with larval hemocytes. This was partially reversed in a dose dependent manner with S methyl thiourea (SMT), a specific inhibitor of inducible nitric oxide synthase (iNOS). A decrease in NO production was also observed when non-selective inhibitors such as N(G)-nitro-L-arginine (L-NAME) and N-omega-nitro-L-arginine (L NNA) were used, albeit the inhibition was not to the extent of SMT. Challenge with the entomopathogenic Gram-negative bacterium Photorhabdus asymbiotica also enhanced NO production by hemocytes of both stages. SMT, alone or in combination with P. asymbiotica significantly decreased levels of NO production. However, it was observed that phenoloxidase activity (a cascade for innate immune responses) was independent of NO production stimulation. NO donors, S-nitroso-N-acetyl penicillamine (SNAP) and diethylenetriamine NO adduct (DETA/NO) at various concentrations (100-500 microM) resulted in the lysis of hemocytes dose dependently. The nitrite production in these cases was however similar to LPS stimulation (10 microg/mL) and 1.5-3 fold lower than those observed upon P. asymbiotica (2.5 x 10(7) cfu/mL) stimulation. Survival analysis (Kaplan-Meier) following injection of P. asymbiotica alone or in combination with SMT revealed that only 12.5% (median survival 25.5 h) of co-injected larvae of G. mellonella survived in comparison to 28.6% (median survival 29 h) survivors in P. asymbiotica alone-injected groups till the end of the study. In contrast, co injected pharate prepupa survived longer (median survival 28 h) than the P. asymbiotica alone-injected individuals (median survival 24 h); however, both co injected and P. asymbiotica-injected groups showed 100% mortality at the end of the study. Based on the above, we propose that although NO production is involved in cellular immune responses of this insect to bacterial infection it does not appear to be a part of the signalling pathway that initiates the prophenoloxidase (PPO) cascade, and the extended NO production/overproduction by pharate prepupal hemocytes could result in cytotoxic rather than cytoprotective effects compared with larval hemocytes. PMID- 16316787 TI - A beta-galactose-specific lectin isolated from the marine worm Chaetopterus variopedatus possesses anti-HIV-1 activity. AB - A 30 kDa beta-galactose-specific lectin named CVL was isolated from the polychaete marine worm Chaetopterus variopedatus (Annelida) and its anti-HIV-1 activity in vitro was determined. Results showed that CVL inhibited cytopathic effect induced by HIV-1 and the production of viral p24 antigen. The EC(50) values were 0.0043 and 0.057 microM, respectively. Time-of-addition analysis of anti-HIV-1 activity indicated its action was at the early stage of virus replication. CVL could blocked the cell-to-cell fusion process of HIV infected and uninfected cells with an EC(50) of 0.073 microM. The inhibition of HIV-1 entry into host cells was demonstrated by using fluorescence-based real-time quantify PCR. At CVL concentration of 0.33 microM and 0.07 microM, 86% and 21% virus attachment were blocked, respectively. The anti-HIV-1 action of CVL might relate to blockade of HIV-1 entry into cells. PMID- 16316788 TI - Center for clinical studies in a surgical department--an approach for more evidence-based medicine. AB - The concept of evidence-based medicine (EBM) has been introduced into the field of surgery in Germany since 1996, initially focussing on the application of the classical five-step-concept of EBM to the individual patient. Despite the international efforts of the Cochrane Collaboration, few systematic reviews address surgical procedures and the number of high quality randomized-controlled trials (RCTs) are low. External evidence in surgery is a major problem and requires new strategies in research and patient care. Therefore, in the Department of Surgery at the University of Heidelberg a clinical trial unit was founded to create more evidence via two approaches. First, patients admitted to hospital should be treated within RCTs. Since May 2002 1808 patients have been enrolled into a total of 38 pharmaceutical (19 trials with 470 patients enrolled) and investigator initiated (19 trials with 1338 patients enrolled) studies. Second, the clinical site develops and conducts surgical efficacy RCTs and follows new standards including publication and registration of the protocols. Within 3 years it was possible to change practice in an academic setting from the traditional concept to an evidence-based approach. PMID- 16316789 TI - Enriched analgesic efficacy studies: an assessment by clinical trial simulation. AB - BACKGROUND AND OBJECTIVE: Enrichment strategies which select subjects who appear to respond to the drug have been used in drug studies to demonstrate clinical efficacy. We have used clinical trial simulation techniques to examine factors that are relevant in clinical trial design based on enrichment where poor responders are excluded from the double-blind phase of the study. METHODS: Simulations were performed for an analgesic trial design involving an open-dose titration phase (enrichment phase) followed by a double-blind, randomized, placebo-controlled maintenance phase. Enrichment was examined by excluding subjects above a predefined pain score (cutoff) from analysis of efficacy for the maintenance phase. Cutoff pain scores ranging from 4 to 7 on a 0 to 10 categorical scale were examined. A database consisting of chronic pain patients who participated in studies with a new formulation of buprenorphine was used to build the simulation model. Since no data were available for the key model variable "correlation between treatment and placebo response", values of 0.25, 0.5, and 0.75 were used for the simulations. RESULTS: A correlation between treatment and placebo effect ranging from 0.75 to 0.25 will cause the likelihood of trial success to vary from 50% to 95%. This model also shows that recruitment efficiency will decrease with the use of lower cutoff pain scores. CONCLUSION: Prior to using enrichment techniques, investigators must consider the correlation between treatment effect and placebo response to optimize clinical trial design. PMID- 16316790 TI - TNFalpha and PTH utilize distinct mechanisms to induce IL-6 and RANKL expression with markedly different kinetics. AB - Parathyroid hormone (PTH) and tumor necrosis factoralpha (TNFalpha) are bone resorptive agents that upregulate interleukin-6 (IL-6) and RANKL production by osteoblasts. IL-6 mRNA expression induced by PTH is rapid and transient in osteoblasts both in vitro and in vivo. This study found that IL-6 secretion induced by PTH is also rapid and transient. The induction of RANKL mRNA by PTH is also rapid and transient although with an extended time course compared to that of IL-6 mRNA. In contrast, the effects of TNFalpha are biphasic. During the first 2 h of stimulation with TNFalpha, the responses are similar to those induced by PTH. This is followed by a period of relatively low IL-6 and RANKL mRNA levels and little IL-6 secretion. A late phase of increased IL-6 and RANKL mRNA expression occurs 12-24 h after stimulation with TNFalpha leading to a significant increase in IL-6 secretion. A similar biphasic pattern of activation of p38 MAP kinase is induced by TNFalpha. p38alpha/beta activation is required for the increased RANKL mRNA during the early phase of stimulation by TNFalpha but not in the late phase. In contrast, p38alpha/beta activation is not required for increased IL-6 mRNA or IL-6 protein secretion in either the early or late phases of stimulation by TNFalpha. Blocking the increases in IL-6 transcription completely eliminates IL-6 secretion induced during the early phases of stimulation by either PTH or TNFalpha. Consistent with the dependence on transcription, IL-6 mRNA is rapidly degraded with half-lives of 10-14 min following stimulation with either PTH or TNFalpha. In contrast to IL-6, RANKL mRNA is substantially more stable with half-lives of 40-60 min. Taken together, our results show that TNFalpha and PTH utilize distinct mechanisms to induce IL-6 and RANKL expression with markedly different kinetics. The more extensive effect of TNFalpha likely reflects that TNFalpha stimulates IL-6 production and bone resorption in pathological situations. In contrast, the less extensive effect of PTH likely reflects that it acts in physiological situations where it is important to minimize the potential adverse effects of high levels of IL-6 on bone and/or surrounding tissues. PMID- 16316792 TI - The adolescent and substance abuse: current concepts. PMID- 16316793 TI - An experimental study of the testing of occlusal patterns on the same denture bases. 1952. PMID- 16316794 TI - Replacement of two mandibular central incisors using a zirconium resin-bonded fixed partial denture: a clinical report. AB - Maxillary and mandibular anterior tooth loss may cause esthetic and functional problems. Successful treatment for these situations may necessitate a minimally invasive technique to preserve sound remaining abutment tooth structure. This article describes the use of zirconium resin-bonded fixed partial denture as a conservative treatment option for the replacement of 2 mandibular central incisors. PMID- 16316795 TI - The use of all-ceramic restorations to splint periodontally compromised teeth: a clinical report. AB - This clinical report describes the use of an all-ceramic system to restore periodontally compromised teeth. A metal-free post-and-core system and all ceramic crowns were used to splint the teeth and fulfill the esthetic demands. PMID- 16316796 TI - The use of a resin-bonded prosthesis while maintaining the diastemata: a clinical report. AB - A dental arch with diastemata and missing lateral incisors represents a challenge because of discrepancies in tooth size. If an implant-supported prosthesis is not possible or not selected as a treatment option, a modified resin-bonded fixed partial denture may be a treatment option to eliminate the esthetic problems of conventional fixed partial dentures. This clinical report describes the procedures to replace congenitally missing lateral incisors with a resin-bonded prosthesis, while maintaining diastemata. PMID- 16316797 TI - Immediate dentin sealing improves bond strength of indirect restorations. AB - STATEMENT OF PROBLEM: Delayed dentin sealing is traditionally performed with indirect restorations. With this technique, dentin is sealed after the provisional phase at the cementation appointment. It was demonstrated that this chronology does not provide optimal conditions for bonding procedures. Immediate dentin sealing (IDS) is a new approach in which dentin is sealed immediately following tooth preparation, before making the impression. PURPOSE: The purpose of this study was to determine whether there were differences in microtensile bond strength to human dentin using IDS technique compared to delayed dentin sealing (DDS). MATERIAL AND METHODS: Fifteen freshly extracted human molars were obtained and divided into 3 groups of 5 teeth. A 3-step etch-and-rinse dentin bonding agent (DBA) (OptiBond FL) was used for all groups. The control (C) specimens were prepared using a direct immediate bonding technique. The DDS specimens were prepared using an indirect approach with DDS. Preparation of the IDS specimens also used an indirect approach with IDS immediately following preparation. All teeth were prepared for a nontrimming microtensile bond strength test. Specimens were stored in water for 24 hours. Eleven beams (0.9 x 0.9 x 11 mm) from each tooth were selected for testing. Bond strength data (MPa) were analyzed with a Kruskal-Wallis test, and post hoc comparison was done using the Mann-Whitney U test (alpha=.05). Specimens were also evaluated for mode of fracture using scanning electron microscope (SEM) analysis. RESULTS: The mean microtensile bond strengths of C and IDS groups were not statistically different from one another at 55.06 and 58.25 MPa, respectively. The bond strength for DDS specimens, at 11.58 MPa, was statistically different (P=.0081) from the other 2 groups. Microscopic evaluation of failure modes indicated that most failures in the DDS group were interfacial, whereas failures in the C and IDS groups were both cohesive and interfacial. SEM analysis indicated that for C and IDS specimens, failure was mixed within the adhesive and cohesively failed dentin. For DDS specimens, failure was generally at the top of the hybrid layer in the adhesive. SEM analysis of intact slabs demonstrated a well-organized hybrid layer 3 to 5 microm thick for the C and IDS groups. For DDS specimens the hybrid layer presented a marked disruption with the overlying resin. CONCLUSIONS: When preparing teeth for indirect bonded restorations, IDS with a 3-step etch-and rinse filled DBA, prior to impression making, results in improved microtensile bond strength compared to DDS. This technique also eliminates any concerns regarding the film thickness of the dentin sealant. PMID- 16316798 TI - Finite element analysis of mechanism of cervical lesion formation in simulated molars during mastication and parafunction. AB - STATEMENT OF PROBLEM: The mechanical theory of cervical lesion formation is popular; however, the mechanism of formation of these lesions is not fully explained. PURPOSE: The aim of this study was calculation of the stresses and Tsai-Wu strength ratio in the cervical area of the mandibular molar during grinding, clenching, and mastication, as well as theoretical investigation of the mechanism of cervical lesion formation in teeth. MATERIAL AND METHODS: A 2 dimensional finite element model of the mandibular first molar and crown of the opposing maxillary molar in the frontal section was developed. Computational simulation of mastication of a bolus with high elastic modulus, including grinding and clenching, was performed. Pairs of contact elements were used between the bolus and occlusal surfaces of the teeth. The analysis was nonlinear. During these simulations, the pressure exerted on the occlusal surface and the state of stresses in the mandibular molar were calculated. To evaluate the strength of anisotropic tooth tissues, the Tsai-Wu failure criterion was applied. This criterion considers the difference in strength of materials due to tensile, compressive, and shear stresses. RESULTS: Significant pressures were exerted on lingual cusps of the mandibular molar model during computer simulations of physiological and pathological load. In enamel elements close to the buccal cemento-enamel junction (CEJ) of the studied tooth, tensile stresses were observed which exceeded the strength of the enamel. In this area, the Tsai-Wu strength ratio reached values higher than 1. According to the Tsai-Wu criterion, these elements were damaged and, thus, were removed from the computer tooth model. During subsequent modeling of the tooth with the initiated cervical lesion, the Tsai-Wu ratio exceeded 1 along the dentino-enamel junction (DEJ), creating an overhang of enamel in the cervical area. Application of minimal horizontal force caused a fracture of this fragile, unsupported enamel fragment. CONCLUSIONS: Overloading of theoretical teeth by computer simulation resulted in enamel damage at the CEJ and led to initiation of a cervical lesion. Subsequent overloading resulted in enamel destruction along the DEJ. The overhanging enamel fragment may easily be chipped. This process was repeated during subsequent tooth overloading and caused enlarging of the lesion. PMID- 16316799 TI - An analysis of maxillary anterior teeth: facial and dental proportions. AB - STATEMENT OF PROBLEM: The size and form of the maxillary anterior teeth are important in achieving pleasing dental and facial esthetics. However, little scientific data have been defined as criteria for evaluating these morphological features. PURPOSE: This study analyzed the clinical crown dimensions of maxillary anterior teeth to determine whether consistent relationships exist between tooth width and several facial measurements in a subset of the Turkish population. MATERIAL AND METHODS: Full-face and anterior tooth images of 100 Turkish dental students viewed from the front and engaged in maximum smiling were recorded with digital photography under standardized conditions. Gypsum casts of the maxillary arches of the subjects were also made. The dimensions of the anterior teeth, the occurrence of the golden ratio, the difference between the actual and perceived sizes, and the relationship between the anterior teeth and several facial measurements by gender were analyzed using the information obtained from both the computer images and the casts. One-sample, 2-sample, and paired t tests, and repeated-measures analysis of variance and Duncan multiple-range tests were performed to analyze the data (alpha=.05). RESULTS: The dimensions of the central incisors (P<.05) and canines (P<.01) varied by gender. The existence of the so called "golden proportion" for the maxillary anterior teeth as a whole was not found. Significant differences emerged when the mean ratios between various perceived widths were compared with their ideal golden ratios (P<.01). Proportional relationships between the bizygomatic width and the width of the central incisor, and the intercanine distance and the interalar width in women were observed. CONCLUSION: The maxillary central incisor and canine dimensions of men were greater than those of women in the Turkish population studied, with the canines showing the greatest gender variation. Neither a golden proportion nor any other recurrent proportion for all anterior teeth was determined. Bizygomatic width and interalar width may serve as references for establishing the ideal width of the maxillary anterior teeth, particularly in women. PMID- 16316800 TI - The effect of investment material type on the contamination zone and mechanical properties of commercially pure titanium castings. AB - STATEMENT OF PROBLEM: Different types of investment materials affect the formation of a surface contamination zone within commercially pure titanium (cpTi) castings. This contamination zone may possibly alter the mechanical properties of cast titanium, which may be problematic for castings used in the fabrication of removable and fixed prostheses. PURPOSE: The purpose of this study was to evaluate the effect of different types of investments on the extent of contamination zone and the modulus of elasticity, yield strength, elongation, and hardness of cpTi castings. MATERIAL AND METHODS: Forty wax patterns were fabricated according to ISO 9693 for tensile testing. The patterns were divided into 2 groups of 20 patterns each, invested, and cast in pairs using cpTi. The first group (P) was invested with a phosphate-bonded silica-based investment material (Ticoat S+L), and the second group (M), with a magnesia-alumina investment material (Rematitan Ultra). Investment materials were examined by x ray diffraction analysis (XRD). One specimen from each group was sectioned and prepared for metallographic observation. The extent of the contamination zone was determined by scanning electron microscopy, using back-scattering electron imaging and energy dispersive spectroscopy analysis, as well as microhardness testing. The tensile strength of the specimens was determined in a universal testing machine. From the derived tensile curves, the modulus of elasticity, yield strength, and percentage elongation were calculated and statistically evaluated among the groups using the Student t test (alpha=.05). Three fractured specimens from each group were examined by scanning electron microscopy to determine the mode of fracture. RESULTS: XRD analysis showed that silica and magnesia were the dominant phases of Ticoat S+L and Rematitan Ultra, respectively. The contamination zone was found to extend 50 to 80 mum for the P specimens and 15 to 20 mum for the M specimens. No significance difference was found for the modulus of elasticity (P=85 +/- 11 GPa, M=79 +/- 13 GPa), whereas significant differences were found for the yield strength (P=462 +/- 48 MPa, M=321 +/- 54 MPa; P<.001) and percentage elongation (P=12 +/- 2, M=21 +/- 7; P=.002) between the groups tested. The fracture mode was brittle externally and ductile internally for both groups. CONCLUSIONS: According to the results of this study, the extent of the contamination zone as well as the yield strength and percentage elongation of the cpTi castings were significantly affected by the type of the investment material. PMID- 16316801 TI - Influence of polymerization mode on flexural properties of esthetic resin luting agents. AB - STATEMENT OF PROBLEM: Dual-polymerized esthetic resin luting agents have become popular. However, it is not clear whether the autopolymerized versions of such products have adequate strength to meet clinical requirements. PURPOSE: The aim of this study was to investigate the effect of different polymerization modes on the flexural properties of esthetic resin luting agents. MATERIAL AND METHODS: Ten esthetic resin luting agents were studied. Materials with 3 polymerization modes (dual, auto, and light) were Appeal, Calibra, Illusion, Lute-It, and Variolink-II; those with 2 polymerization modes (dual and auto) were Bistite-II DC, Cement-It-C&B, Clearfil-DC-Cement, Linkmax, and Nexus2-Dual-Syringe. Five flexural strength specimens (2 x 2 x 25 mm) were made for every available polymerization mode for each material. The specimens were stored in distilled water for 24 hours at 37 degrees C. The specimens were then tested for flexural strength (MPa) and flexural modulus (GPa) using the 3-point bending method on a universal material testing machine at a cross-head speed of 0.5 mm/min. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (alpha=.05). RESULTS: Flexural strength values were the highest for dual-polymerized Nexus2-Dual-Syringe (155 MPa), whereas the values were lowest for autopolymerized Bistite-II-DC (56 MPa) and light-polymerized Appeal (63 MPa). Flexural moduli values ranged from 4.3 to 10.0 GPa. The polymerization mode and luting agent influenced flexural strength and modulus significantly (P<.05). CONCLUSION: Dual-polymerized resin luting agents had higher or equal flexural strength compared to the autopolymerized mode. All the groups tested passed the flexural strength requirement of the ISO 4049 specification. PMID- 16316802 TI - Evidence-based considerations for removable prosthodontic and dental implant occlusion: a literature review. AB - The dental literature is filled with discussions of dental occlusion, occlusal schemes, philosophies, and methods to correct and restore the diseased, worn, or damaged occlusion. Traditionally, these discussions have been empirical in nature and not based on scientific evidence. Due to the empirical nature of the literature, the study of occlusion has been extremely complex and troublesome to both pre- and post-doctoral students. The introduction of osseointegrated implants has further complicated the situation. Dentists may apply the principles of occlusion for the natural dentition directly to implant-supported and retained restorations. Although this may be successful, this rationale may result in overly complex or simplified treatment protocols and outcomes. There is an emerging body of scientific literature related to dental implant therapy that may be useful in formulating treatment protocols and prosthesis designs for implant supported restorations. This review focuses on some of the "classic" removable prosthodontic literature and the currently available scientific literature involving removable prosthodontic occlusion and dental implant occlusion. The authors reviewed the English peer-reviewed literature prior to 1996 in as comprehensive manner as possible, and material after 1996 was reviewed electronically using MEDLINE. Electronic searches of the literature were performed in MEDLINE using key words-animal studies, case series, clinical trials, cohort studies, complete denture occlusion, dental implant function, dental implant occlusion, dental implant occlusion research, dental implant functional loading, dental implants, dental occlusion, dental occlusion research, denture function, denture occlusion, dentures, implant function, implant functional loading, implant occlusion, occlusion, and removable partial denture occlusion-in various combinations to obtain potential references for review. A total of 5447 English language titles were obtained, many of which were duplicates due to multiple searches. Manual hand searching of the MEDLINE reference list was performed to identify any articles missed in the original search. PMID- 16316803 TI - A new mathematical process for the calculation of average forms of teeth. AB - STATEMENT OF PROBLEM: Qualitative visual inspections and linear metric measurements have been predominant methods for describing the morphology of teeth. No quantitative formulation exists for the description of dental features. PURPOSE: The aim of this study was to determine and validate a mathematical process for calculation of the average form of first maxillary molars, including the general occlusal features. MATERIAL AND METHODS: Stone replicas of 174 caries free first maxillary molar crowns from young patients ranging from 6 to 9 years of age were measured 3-dimensionally with a laser scanning system at a resolution of approximately 100,000 points. Then, the average tooth was computed, which captured the common features of the molar's surface quantitatively. This new method adapts algorithms both from computer science and neuroscience to detect and associate the same features and same surface points (correspondences) between 1 reference tooth and all other teeth. In this study, the method was tested for 7 different reference teeth. The algorithm does not involve any prior knowledge about teeth and their features. RESULTS: Irrespective of the reference tooth used, the procedure yielded average teeth that showed nearly no differences (less than +/-30 microm). CONCLUSION: This approach provides a valid quantitative process for calculating 3-dimensional (3D) averages of occlusal surfaces of teeth even in the event of a high number of digitized surface points. Additionally, because this process detects and assigns point-wise feature correspondences between all library teeth, it may also serve as a basis for a more substantiated principal component analysis evaluating the main natural shape deviations from the 3D average. PMID- 16316804 TI - Die pin channel cleaner. PMID- 16316805 TI - A method to make an interocclusal record for mandibular advancement devices. PMID- 16316806 TI - A technique for the manipulation of prosthetic screws intraorally. PMID- 16316807 TI - An alternative technique for fabricating a single-implant-supported crown. PMID- 16316809 TI - Nutrition and primary prevention of breast cancer: foods, nutrients and breast cancer risk. AB - Worldwide, each year approximately one million women are newly diagnosed with breast cancer (BC), in Germany 65 new cases per 100,000 inhabitants are registered, yearly. The fact that incidence has been rising in parallel with economic development indicates that environmental factors might play a role in the causation of BC. Migrational data have pointed to nutrition as one of the more relevant external factors involved. Preventive dietary advice often includes a reduction of alcohol, red meat and animal fat and increasing the intake of vegetables, fruit and fibre and lately, phyto-estrogens from various sources. Clearly, the scientific basis for these recommendations appears sparse. The available prospective data from epidemiological studies and interventional trials do not support the overall hypothesis that higher fat-intakes are a relevant risk factor for BC development, more important seems the relative distribution of various fatty acids. A non-vegetarian eating habit (consumption of animal products) per se does not elevate BC risk, while consumption of broiled or deep fried meats cannot be ruled out as a risk factor in genetically susceptible individuals. It appears prudent to abstain from regular and increased alcohol consumption. This should be particularly true for pubescent girls, in whom glandular breast tissue is particularly vulnerable. In general, if alcohol is consumed on a regular basis, a sufficient supply of fresh vegetables and fruit is essential. While there is no overall protective effect of a high fruit and vegetable consumption speculation remains over possible beneficial effects of certain subcategories, especially brassica vegetables like broccoli, cauliflower and cabbage. In essence, regional differences in BC incidence are probably partially attributable to life long dietary habits. There is no need to adopt a foreign dietary plan in order to protect oneself against BC. Traditional western diets also have their beneficial ingredients that should be regular constituents in our meals. Lignans from traditionally made sourdough rye bread, linseed/flaxseed and berries are local sources of potentially canceroprotective phyto-estrogens. Furthermore, indole-3-carbinol rich cabbage species might contribute to BC protection by diet. Nevertheless, clear cut recommendations for or against single nutrients or secondary plant metabolites are not yet possible, lacking sufficient data on individual bioavailability, safety and long term outcome. BC prevention by dietary means therefore relies on an individually tailored mixed diet, rich in basic foods and traditional manufacturing and cooking methods. PMID- 16316810 TI - Teratogenic potential of vaginal metronidazole treatment during pregnancy. AB - OBJECTIVE: To study the human teratogenic potential of vaginal metronidazole treatment during pregnancy. DESIGN AND SETTING: The analysis of cases with different congenital abnormalities and their matched controls in the population based data set of the large Hungarian Case-Control Surveillance of Congenital Abnormalities (HCCSCA), 1980-1996. STUDY PARTICIPANTS: Pregnant women (38,151) who had newborn infants without any congenital abnormalities (control group), 22,843 pregnant women who had newborn infants or fetuses with congenital abnormalities. MAIN OUTCOME MEASURES: Twenty-two congenital abnormality groups. RESULTS: The prevalence of vaginal metronidazole treatment during pregnancy was 1.7% in the case group (388 pregnant women), while it was 1.5% in the control group (570 pregnant women) [crude prevalence odds ratio (POR) with 95% confidence interval (CI): 1.1, 1.0-1.3]. Comparisons of cases and their matched controls showed an association between vaginal metronidazole treatment during the second and third months of gestation and congenital hydrocephalus (adjusted POR with 95% CI: 10.7, 1.1-104.5), however, based only on five cases. The evaluation of medically recorded metronidazole treatment did not confirm this association. CONCLUSION: Our finding can only be regarded as a signal for the possible association between vaginal treatment with metronidazole during pregnancy and congenital hydrocephalus. PMID- 16316811 TI - Predictive value of glucose-insulin ratio in PCOS and profile of women who will benefit from metformin therapy: obese, lean, hyper or normoinsulinemic? AB - OBJECTIVE: The aim of the present study is to evaluate sub-groups of PCOS patients who will benefit from metformin therapy and to find out any predictors of ovulation in PCOS sub-groups. METHOD: In the current prospective-randomized, placebo-controlled, double-blind study, PCOS patients (n = 116) were divided into six main groups according to glucose to insulin ratio (G-I ratio mg/10(-4) U) and body mass index (BMI kg/m2) as: Group 1: normoinsulinemic (G-I ratio > or = 4.5 mg/10(-4) U), lean (BMI < 25) (n = 37); Group 2: normoinsulinemic, overweight (BMI: 25-29.9) (n = 19); Group 3: normoinsulinemic, obese (BMI > or = 30) (n = 18); Group 4: hyperinsulinemic (G-I ratio < 4.5 mg/10(-4) U), lean (n = 28); Group 5: hyperinsulinemic, overweight (n = 17); Group 6: hyperinsulinemic, obese (n = 20). Patients in each group were randomized onto placebo or metformin treatments (850 mg two to three times per day according to BMI). The rate of ovulation, biochemical profile, hormonal profile and clinical symptoms of hyperandrogenism were evaluated before and after 6 months of metformin and placebo treatments. RESULT(S): We observed a significant decrease in WHR following metformin therapy in the normoinsulinemic overweight sub-group (P < 0.05). The duration of the menstrual cycle significantly decreased in the normoinsulinemic obese sub-group on metformin therapy (P < 0.05). Metformin had a significant effect on hirsutism scores in hyperinsulinemic lean women (P < 0.05) and decreased DHEAS levels significantly in the lean hyperinsulinemic and normoinsulinemic groups (P < 0.05). Metformin had significant effects on ovulation in only lean hyperinsulinemic women (P < 0.05). CONCLUSIONS: Clinical outcomes of metformin therapy may be categorized on the basis of basal BMI and insulin levels in PCOS patients. PMID- 16316812 TI - Antiphospholipid syndrome during pregnancy: associated with early onset of HELLP syndrome and liver infarctions. PMID- 16316813 TI - Specialist life--Aris Antsaklis. PMID- 16316815 TI - [Psychopathology of stroke]. AB - The psychopathology of stroke encompasses several psychiatric and behavioral disorders that have high prevalence in the geriatric population, reduce the patient autonomy and increase the caregiver's burden. These disorders are usually associated with other cognitive and neurological deficits, and are labelled as neuropsychiatric when the whole clinical picture is consistent with the specific dysfunction of a neural system or brain region. Thus the neuropsychiatry of stroke comprises disorders of the perception/identification of the self and the environment (anosognosia of hemiplegia, misidentification syndromes, confabulations, visual hallucinations, delirium and acute confusional state), amotivational syndromes (apathy and athymhormia), disorders of emotional reactivity (blunted affect, emotional incontinence, irritability, catastrophic reactions), poor impulse or ideation control (mania) and personality changes. The clinical profile of the subcortical vascular dementia also points to specific brain dysfunction (frontal-subcortical pathways) that manifests with behavioral (depression, emotionalism, irritability) and cognitive symptoms (psychomotor retardation, attention, executive and memory deficits). However, post-stroke depression and anxiety, which have a more variable clinical presentation and might be assimilated, for several aspects, to post-traumatic or adaptive disorders, are disorders less characterized in their neural correlates. PMID- 16316816 TI - [Mixed dementia: a neuropathologic point od view]. AB - Alzheimer's disease (AD) and vascular dementia (VaD) are the most frequent causes of dementia in the elderly. Although AD can be diagnosed with a very high degree of accuracy, the distinction between pure AD, VaD and mixed dementia (MD), where both pathologies co-exist in the same patient, remains a controversial issue and one of the most difficult diagnostic challenges. MD represents a very frequent pathology, especially in the elderly, as underlined by the neuropathological studies. However, the respective importance of degenerative and vascular lesions, their interaction in the genesis of dementia and the mere existence of mixed dementia are still debated. Accurate diagnosis of MD is of crucial significance for epidemiologic purposes and for preventive and therapeutic strategies. Until recently, pharmacological studies have generally focused on pure diseases, either AD or VaD, and have provided little data on the best therapeutic approach to MD. This review will provide an overview of neuropathological aspects of MD in the elderly, which appears to be one of the most common forms of dementia. PMID- 16316817 TI - [Animal models of Alzheimer's disease: a road full of pitfalls]. AB - Alzheimer's disease (AD) is a neurodegenerative disorder that affects people slowly, insidiously, progressively but irreversibly. This disease will destroy, little by little, the neurons of the hippocampal formation that sustain episodic memory, and the neurons of the polymodal association areas involved in all other cognitive functions. AD is characterized by two types of brain lesions: amyloid plaques and neurofibrillary degeneration. Three major molecular actors are involved in the dynamic of neurodegeneration, but the precise role of each is still a matter of debate: the first one is APP (amyloid protein precursor) that is cleaved to release Abeta peptide that will aggregate into plaques. The last one is the microtubule-associated protein tau that assembles into paired helical filaments in neurons to constitute neurofibrillary degeneration. The main difficulty to study AD results from the fact that this disease is specific to humans and, therefore, that there is no relevant animal model at our disposal. Transgenic mice merely reflect partial aspects of the physiopathological process, impeding therapeutic approaches such as relevant drug tests on animals. But research is in progress... PMID- 16316818 TI - [Dementia and falls: two related syndromes in old age]. AB - Dementia and cognitive impairment are known as a major risk for falls and subsequent adverse events in the elderly. In addition to result in serious injury, including fractures, falls lead to functional decline due to fear of falling again and self limitation of activity in older adults. All types of dementia and all degrees of severity are involved. Rather than resulting from a single cause, falls are the result of a combination of intrinsic, situational, and environmental factors. The most common risk factors for falls in patients with cognitive impairment and dementia are gait and balance disturbances, behavioral disorders, visual problems, malnutrition, adverse effects of drugs, fear of falling, neurocardiovascular instability (particularly orthostatic hypotension), and environmental hazards. Based on data from studies in cognitively normal people who fall, a multifaceted intervention, including a physical exercise programme and modification of the risk factors may prevent falls in older people with cognitive impairment and dementia. Preliminary research suggests that physiotherapy may have a role for falls prevention in these patients. However, randomized studies need to be performed. PMID- 16316819 TI - [Sexuality and dementia]. AB - Sexuality, love, companionship and intimacy remain important parts of life in older people and demented patients. The most frequent sexual disorder reported by spouses of patients with dementia is sexual indifference related to apathy and blunted affect. Increase of sexual demands is rare and many behaviors considered as inappropriate or expressing hypersexuality actually express affective needs or result from cognitive disturbances. Permanence of sexual activity is most often a factor of adjustment for married dementia sufferers and their caregivers. However, some sexual behaviors are stressing for the spouses, mainly women spouses. In nursing homes or long term care facilities, expressing sexuality by demented subjects and dealing with inappropriate sexual expression are source of concerns for the nursing staff, other residents, and families. Information about sex and dementia and a psychobehavioral approach can decrease the strain of families and caregivers. PMID- 16316820 TI - [post-traumatic stress disorder in the elderly]. AB - Diagnosis of post traumatic stress disorder (PTSD) requires a past history of psychic traumatism and characteristic psychotraumatic symptoms like re experiencing of the traumatic event, avoidance of stimuli associated with the traumatic event and increased arousal. In the elderly, PTSD prevalence rate is about 0.9% after 60 years of age. Clinical features are almost identical to those observed in younger patients. However there are various types of PTSD: PTSD de novo, occurring after exposure to extreme trauma in old age; chronic PTSD when symptoms persist since the time of the trauma; delayed-onset PTSD when patients exhibit signs of the disorder decades after the trauma; complex PTSD, generated by exposure to repeated traumatisms in early development, who could be linked up to changes in personality in older exposed to repeated traumatisms. Pharmacological and psychotherapic interventions used in younger populations can be applied for treatment to older adults, when taking into account the pharmacodynamic and psychic changes associated with aging. PMID- 16316821 TI - [Forgetting: a product of memory]. AB - If someone asks you what your children talk about at breakfast this morning, it is likely that you not able to tell him. This forgetting will not appear like a mark of memory disorder. But if it concerns a 80 years old people! Over the last decade, it has been claimed that aging induces reduced inhibitory processing and this has been put forward to explain forgetting. However, several experiments have proposed that non inhibitory mechanisms can account for the pattern of results. So, aging doesn't involve inhibitory deficit. Recently, a procedure known as the "retrieval practice paradigm," claimed to give a good measure of inhibitory processing whose goal is to suppress competing traces during an episodic retrieval. In this paradigm, participants study exemplars from taxonomic categories (e.g., fruit-orange, fruit-banana, drink-scotch) and then practice retrieving half of the exemplars from half of the categories by recalling those studied items given a category and letter stem as cue (e.g., fruit-or _). Each item is tested several times. After a delay, subjects are tested on all items. Practice impairs recall of the remaining non practiced exemplars of the practiced categories, phenomenon known as "retrieval-induced forgetting". In the present experiment, we would to determine whether retrieving an item from semantic memory inhibits other concepts that compete with it during retrieval. Indeed, studies of lexical ambiguity resolution provide evidence for inhibition in semantic retrieval. For that, we use homographs because their natural characteristic involves inhibiting one of their two meanings. We expected that if participants practice a word related to a homograph's subordinate meaning, word related to its dominant meaning would need to be inhibited. Results show that old people showed normal levels of inhibition. Moreover, results show an interaction between practiced-unpracticed items and homograph's polarity. These results suggest that a deficit in inhibitory process can not explain forgetting. PMID- 16316822 TI - Ethical issues in pediatric oncology phase I-II trials based on a mother's point of view. AB - Phase I-II trials are developing in Pediatrics and raise many complex relational, psychological and ethical issues. We present and discuss these based on an interview in a pediatric oncology setting, with a mother who accepted that her daughter be included in such trials and who expressed why she accepted with great sensitivity and profoundness. She explained that after many years of inefficient treatments she had lost all her landmarks and was ready to accept any proposition, even those she would have considered unacceptable earlier. She did not know whether there is a limit to what is acceptable. Her only objective was to gain any time possible in order to continue living with her daughter. She found it important that the research doctor be different from the doctor involved in patient care, and that the latter remains the major decision-maker and correspondent: thus the child's best interests take precedence over that of research. Interviews with the psycho-oncologist can help the parents and the doctors gain a better insight into the various aspects, rational and irrational, conscious and unconscious, involved in the proposition to participate in a clinical trial and in the parents' or the child's acceptance or refusal. PMID- 16316823 TI - Proteasome inhibition: a new approach for the treatment of malignancies. AB - Since last years, the proteasome has emerged as a real and exciting target for anticancer therapy. Velcade (bortezomib, formerly known as PS341) remains the first selective proteasome inhibitor that has demonstrated significant preclinical activity in several tumor models and a significant efficacy in patients with refractory or relapsed multiple myeloma, resulting in an accelerated approval in US and Europe in such a setting. The major biological effect of bortezomib is the inhibition of the nuclear transcription factor NFkappaB, with subsequent inhibition of the growth tumor cells, induction of apoptosis, inhibition of angiogenesis and of cellular adhesion. The better understanding of the role of proteasome in the regulation of tumor cell growth has led to the development of new therapeutic approaches, notably in patients with multiple myeloma but also seems to hold interesting promises in other hematologic malignancies and solid tumors. This review provides a summary of the rationale for using proteasome inhibitors and an update on available and ongoing clinical studies involving human malignancies. PMID- 16316824 TI - [The 2005 Nobel Prize in Medicine: after stress, bacterial infection]. PMID- 16316825 TI - [The RASSF1 (Ras association domain family protein 1) gene. Category: tumor suppressor gene]. PMID- 16316827 TI - [New insights into cytotoxic effector cells]. AB - Cytotoxic immune cells play a role against infectious or tumour invasion. There are two types of cytotoxic cells : those involved in innate or natural immunity (natural killer cells, Tgammadelta lymphocytes, NKT cells) and those involved in the adaptive or acquired immunity (cytotoxic T lymphocytes). Recent advances in immunology have allowed the characterisation of unconventional cytotoxic populations, to better individualise the real cytotoxic cells within the traditional CD8+ T-cell population, and finally to get more insights into the activation of the cytotoxicity properties, in particular with regard to NK cells. The data recently acquired concerning the effector cytotoxic cells contribute to a better understanding of the interactions between the immune system and tumour cells. This may provide rational basis for combined immunotherapy protocols. PMID- 16316829 TI - [Management of rectal cancer in pregnant women]. AB - We relate 2 cases reports about rectal cancer and pregnancy. This association is rare but is a real problem of management because diagnosis is done latly and it mate have incompatibility between treatments and pregnancy. A medical bibliography has been done, to define the best medical procedure in function of the disease staging and the pregnancy term. It shows that a multi disciplinary decision must be done, which take into consideration the choice of the obstetricals, pediatricians, surgeons, and oncologists, but also the patient's choice. PMID- 16316830 TI - [Asbestos and malignant pleural mesothelioma: molecular, cellular and physiopathological aspects]. AB - Asbestos is known as mutagenic and carcinogenic for human and is responsible for many pulmonary diseases including asbestosis, bronchogenic carcinoma and malignant pleural mesothelioma. Occupational exposure to asbestos is involved in 70-80% of all malignant pleural mesothelioma. The later presents a growing challenge for both researcher and clinician. The diagnosis of malignant pleural mesothelioma is difficult and the current treatments did not show significant improvement of the survival. The increasing incidence of malignant pleural mesothelioma, its gravity and its human, social and financial consequences are of high concern in public health. In this paper we summarize the so far knowledge on cellular, molecular and pathophysiological events involved in genesis and development of malignant pleural mesothelioma. Finally, the paper also report recent data sourced from the study of malignant pleural mesothelioma transcriptome using high-throughput technologies such as gene expression array. These data should improve the accuracy of mesothelioma diagnosis and therapy. PMID- 16316831 TI - [Nasopharyngeal carcinoma in Tunisian children: retrospective epidemiological, clinical and biological study about 48 cases]. AB - OBJECTIVE: Report epidemiological, clinical and biological aspects of nasopharyngeal carcinoma in Tunisian children. PATIENTS AND METHODS: Our retrospective study from 1994 to 2001 included all children treated for nasopharyngeal carcinoma in the Salah Azaiz Cancer Institute of Tunis. Initial investigation consisted of ENT and general examination, nasopharyngeal CT-scan, abdominal echography, chest X-ray and bone scintigraphy. Biological markers included blood-count, erythrocytes sedimentation and serum lactic dehydrogenase. All children received neoadjuvant chemotherapy (adriamycin, cisplatin) and irradiation therapy. RESULTS: There were 48 children with a median age of 13,7 years and a sex ratio of 1,4 (28/20). Lesions are staged T2, T3 and T4 in 2,1 %, 18,7% and 79,2% of cases. All patients have cervical palpable nodes at diagnosis classified as N1 (8,3%), N2 (33.3%) and N3 (58.3%). A significant correlation was found between serum lactic dehydrogenase and the N stage (p = 0.02). After follow up, recurrence of disease was noted for three children, persistent disease for two children and metastatic disease in five cases. The overall and relapse free survival at 5 years were 79.1% and 68.9% respectively. Patients aged 13 or lower had poorer 5 overall survival rate (72.3%) than older age group (84.2%). PMID- 16316832 TI - [Breast adenocarcinoma: critical analysis of sentinel lymph node histopathological results of 542 procedures]. AB - Between February 2001 and March 2003, 542 sentinel lymph node procedures were performed for localised breast carcinoma (T0-T1, N0, M0) without any previous treatment. Frozen sections were performed in 515 cases and they did not reveal metastases in 446 cases. Fifty-two micrometastases < 2 mm and 18 macrometastases were reported by definitive histopathological exam. Axillary clearance was performed in 50/70 patients (38 with micrometastases and 12 for macrometastases). Modalities of histopathological procedure are discussed and particularly number and interval of serial slides with or without immunochemistry ; 81.8% (36/44) of micrometastases were detected on the two first serial sections. Decisional value of axillary clearance performed in case of micrometastases is also evaluated. PMID- 16316833 TI - [Retrospective study of a series of 26 carcinomatous meningitis secondary to lung cancer]. AB - Carcinomatous meningitis occur in approximately 5 to 18% of bronchial carcinoma. We report here the analysis of a retrospective study of 26 cases of carcinomatous meningitis secondary to a bronchial carcinoma. The most frequently involved histological types are adenocarcinoma and small cell lung cancer. The diagnosis is difficult because of the clinical polymorphism and the inconstant presence of neoplasic cells in the cerebrospinal fluid. The prognosis is very poor with a median survival time of 4 weeks. Development of new molecules as well as an earlier diagnosis could contribute to improve the outcome. PMID- 16316834 TI - [Economic evaluation of the new national breast cancer screening programme in France: application to the Bouche-du-Rhone district]. AB - The purpose is to measure the costs of the new national breast cancer screening programme in France and to compare these with those of the previous programme in the Bouches-du-Rhone district. Direct screening costs and costs related to diagnosis and assessment were collected. Costs are presented by screening period, by organisms involved in the screening program and by corresponding phase within the screening process. The total cost of the screening program total cost has increased from 5587487 euros to 9345469 euros between the two campaigns. The main reasons are the investment costs in the new screening program, the increase in the target population and the increased fee for programs. This study presents a first estimate of the costs related to the new national breast cancer screening program. Results of this study may help to guide future decisions on the further development of breast cancer screening in France. PMID- 16316835 TI - [Advanced ovarian cancer: a need for redefining therapeutic objectives and strategy]. PMID- 16316836 TI - [How to consider cancer: implications for the risk due to ionizing radiation]. AB - Cancers induced by ionizing radiation have no particular specificity nor genetic remarkable signature, excepting numerous multideletions. They should therefore be studied in the general field of cancer biology in its broad sense. A gap remains between the initial events like the rather well identified genomic damage and the subsequent emerging cellular clone with cancer characteristics. Intermediate steps are generally described as accumulation of mutations and epigenetic modifications leading at one point to the malignant phenotype. However we have no clear nor understandable model on these steps of malignant transformation till now. It is quite possible that specific causes (tobacco, alcohol, radiations, chemical toxics) which produce different initial abnormalities then lead to (or accelerate the entry in) the common and same way as that resulting of accumulations of damage due to ageing. Genomic instability is certainly an important factor involved in the cellular drift leading to malignant transformation. We postulate that only cells having both a high telomerase activity and a low apoptotic activity may become cancerous. The hypothesis is that cancer results from a genome reprogramming of these cells due to an oncogenic mitotic pressure which induces a loss of the differentiation control. We propose to name anti-apoptosis, in contrast to apoptosis which is the programmed death, the ultimate process by which a cell loses its tissue-related properties. The oncogenic activation may propagate to primordial genes of development resulting in emergence of a subnuclear with tumoral activity. Bio molecular studies of embryonic development and of genome re-programming will probably allow us to better understand the mechanisms of cancer. PMID- 16316838 TI - Sexual dysfunction in women with diabetes mellitus. AB - Sexual problems in men with diabetes mellitus are common and often result from diabetic complications. However, although diabetic complications are similar in both sexes, little attention has been given to the effects of diabetes on female sexuality and sexual function. It is reasonable to suggest that women with diabetes will experience sexual problems due to neuropathy, endocrine changes and vascular complications. However there is little research into the physical effects of diabetes on female sexual function. Reduced vaginal lubrication in women with diabetes has been reported yet this is rarely documented as a sexual problem. Previous studies in women with diabetes may also neglect the importance of the subjective qualities associated with female sexuality and sexual expression. PMID- 16316839 TI - A case-control study of oxidized low density lipoproteins and preeclampsia risk. AB - Diffuse vascular endothelial dysfunction, secondary to oxidative stress, is an important pathological feature of preeclampsia. Oxidative conversion of low density lipoproteins (LDL) to oxidized-LDL (Ox-LDL) is considered an important step in transforming macrophages into lipid-laden foam cells destined to develop into early atherosclerotic-like lesions. In our study of 95 women with preeclampsia and 100 controls, we evaluated the association between maternal plasma Ox-LDL concentrations and preeclampsia risk. Ox-LDL concentrations were measured using a solid phase two-site enzyme immunoassay. Plasma lipids were measured using standard enzymatic procedures. We used logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) adjusted for confounders. Plasma Ox-LDL concentrations were positively correlated with cholesterol, triglyceride (TG), and LDL concentrations in cases and controls, (Spearman's r ranging from 0.39-0.48, p-values all <0.01). There was no evidence of an increased risk of preeclampsia across increasing quartiles of Ox-LDL. The ORs for successive quartiles, with the lowest as the reference group, were as follows: 1.0, 1.1, 0.6, and 1.2. Women with extremely high concentrations of Ox LDL (> or =73 U/L, the upper decile), as compared with those with lower values (<73 U/L) had a 2.7-fold increased risk of preeclampsia (95% CI 1.0-6.8). Women with high Ox-LDL and high TG concentrations (> or =284 mg/dl), as compared with those without these two factors, had a 9.6-fold increased preeclampsia risk (95% CI 2.0-45.6). Elevated Ox-LDL, particularly in conjunction with elevated TG, appears to be a risk factor of preeclampsia. PMID- 16316840 TI - BMI as a modifying factor in the relations between age at menarche, menstrual cycle characteristics, and risk of preeclampsia. AB - To examine the relations between age at menarche and menstrual cycle characteristics and preeclampsia risk, we analyzed data from a case-control study conducted from 1998 to 2002. Usual menstrual cycle characteristics among 286 preeclampsia cases and 471 normotensive controls were assessed using a structured interview during postpartum hospitalization. We used multivariable logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI), adjusting for confounders. We also examined the influence of pre-pregnancy body mass index (BMI) on selected hypothesized relations. Among lean women (pre pregnancy BMI < 25 kg/m(2)), preeclampsia risk was lower in those reporting cycles > or = 36 days in length (adjusted OR 0.78, CI 0.35-1.83) and menarche at > or = 14 years (adjusted OR 0.48, 95% CI 0.28-0.82). In contrast, among overweight or obese women, preeclampsia risk was higher in those with long cycles (OR 3.11, 95% CI 0.62-1.56, p(interaction) = 0.16) and late menarche (OR 1.53, 95% CI 0.59-3.97, p(interaction) = 0.03). These data suggest that adiposity influences the relations between usual menstrual cycle length, age at menarche, and preeclampsia. Overweight women with long cycles are at particularly high preeclampsia risk. PMID- 16316841 TI - Pro12Ala polymorphism of the peroxisome proliferator-activated receptor-gamma gene in first-degree relatives of subjects with polycystic ovary syndrome. AB - AIM: This study was designed to examine the relationship between the Pro12Ala polymorphism of the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) gene and insulin resistance (IR) in first-degree relatives of subjects with polycystic ovary syndrome (PCOS). MATERIALS AND METHODS: One hundred and twenty family members of 55 patients with PCOS and 80 unrelated healthy control subjects without a family history of diabetes or PCOS were studied. IR was assessed by homeostatic model assessment (HOMA-IR) and area under the curve (AUC) for insulin during an oral glucose tolerance test in subjects with normal glucose tolerance and controls. Genetic analysis of the PPAR-gamma gene Pro12Ala polymorphism was performed by restriction fragment length polymorphism. RESULTS: Fasting insulin, HOMA-IR and AUC insulin were significantly higher in first-degree relatives of PCOS subjects than in controls. A significantly different allele distribution of the Pro12Ala polymorphism of PPAR-gamma was observed between the two groups, with the frequency of the variant Ala isoform being significantly reduced in the first degree relatives of PCOS subjects (10.8%, 13 subjects) compared with the control group (22.5%, 18 subjects). All Pro12Ala polymorphisms of the PPAR-gamma gene were heterozygous. Compared with first-degree relatives of PCOS subjects with the Pro12Pro polymorphism of PPAR-gamma, first-degree relatives of PCOS subjects with the Pro12Ala polymorphism had low fasting insulin, HOMA-IR and AUC insulin levels. The combined prevalence rate for impaired glucose tolerance, impaired fasting glucose and diabetes was 40% (16 subjects) in mothers and 52% (20 subjects) in fathers of PCOS women. CONCLUSION: Our findings suggest that Pro12Ala PPAR-gamma gene polymorphism may be protective against IR and might prevent the development of diabetes mellitus in the first-degree relatives of subjects with PCOS. PMID- 16316842 TI - Hepatic lipase gene polymorphism, pre-pregnancy overweight status and risk of preeclampsia among Peruvian women. AB - We examined the association between the hepatic lipase (LIPC) gene promoter polymorphism (-514C/T) and risk of preeclampsia among Peruvian women. We also evaluated whether this association is modified by maternal pre-pregnancy overweight status. Using a case control study design, 157 preeclampsia cases and 180 normotensive controls were enrolled in the study. Genotyping was conducted using PCR amplification, NlaIII enzyme digestion and gel electrophoresis. Logistic regression procedures were used to calculate adjusted odds ratios (ORs) and 95% confidence intervals (CI). After adjusting for confounding by maternal age, parity and pre-pregnancy body mass index (BMI), the relative risks of preeclampsia for women with LIPC -514CT and LIPC -514TT genotypes were 1.0 (95% CI 0.5-2.2) and 1.5 (95% CI 0.7-3.3) respectively, using women with LIPC -514CC genotype as a reference. Women who were both overweight and who had the LIPC 514TT genotype had a significant 3-fold increased risk of preeclampsia (Adj. OR:3.0 95% CI 1.3-6.8) as compared to those women who were not overweight and had the LIPC -514CC/CT genotype. In this study, we found that LIPC -514TT genotype and overweight status, when occurring together, were associated with a 3-fold increase in risk of preeclampsia among Peruvian women. PMID- 16316843 TI - Serum androgen levels in patients undergoing controlled ovarian hyperstimulation for in vitro fertilization cycles. AB - AIM: To investigate androgen behavior during controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF). DESIGN: A prospective, observational study. SETTING: An IVF unit of an academic medical center. PATIENTS AND METHODS: Blood was drawn three times during the COH cycle from 17 consecutive patients undergoing the long gonadotropin-releasing hormone-analog protocol: the day on which adequate suppression was obtained (Day-S); the day of or prior to administration of human chorionic gonadotropin (Day-hCG); and (3) the day of ovum pick-up (Day-OPU). RESULTS: There was a significant increase in serum sex steroid levels during gonadotropin treatment. After hCG administration, there was a significant increase in levels of serum 17-hydroxyprogesterone (17-OHP) and ovarian androgens (total and free testosterone and androstenedione), with no significant change in adrenal androgen (dehydroepiandrosterone sulfate). Significant correlations were observed between plasma estradiol (E(2)) and androgen levels during COH and until hCG administration, but not after hCG administration. The E(2)/testosterone ratio increased significantly during COH and until hCG administration, and then decreased significantly. The number of oocytes retrieved correlated significantly with serum 17-OHP, E(2) and E(2)/testosterone ratio. The number of gonadotropin ampoules used correlated inversely only with serum E(2) levels. CONCLUSION: In patients undergoing COH for IVF, androgen levels increase in response to gonadotropin, and then again after hCG administration. Although the E(2)/testosterone ratio correlates with the number of oocytes retrieved, androgen levels do not. PMID- 16316844 TI - Effects of estrogen replacement therapy on pituitary size, prolactin and thyroid stimulating hormone concentrations in menopausal women. AB - There are few studies in the literature investigating pituitary size at an age consistent with the menopause and the influence of estrogen replacement therapy on pituitary height. We therefore evaluated the effect of estrogen on pituitary size, prolactin and thyroid-stimulating hormone (TSH) levels in menopausal women. Sixty-nine women were evaluated, 47 using estrogen and 22 controls. The measure of pituitary height was obtained from magnetic resonance imaging. Hormone evaluation did not show a statistically significant difference in mean (+/ standard deviation) prolactin level between the group using estrogen (7.6 +/- 6.4 ng/ml) and controls (5.1 +/- 3.4 ng/ml; p = 0.15), yet mean TSH level was significantly higher in the treated group (1.9 +/- 1.5 vs. 1.2 +/- 0.9 microU/ml; p = 0.03). Mean pituitary height in the estrogen-treated group (5.2 +/- 1.4 mm) was greater than in the controls (4.4 +/- 1.4 mm; p = 0.04). However, when such potential confounders as age, prolactin and TSH levels, treatment and duration of estrogen exposure were considered, the magnitude of difference did not attain significance. In conclusion, estrogen may play a role in pituitary size, since a mean difference in pituitary height, estimated as 0.78 mm, was detected between the groups in favor of the estrogen-treated group. This suggestion can be investigated in further studies. PMID- 16316845 TI - A preliminary study on reduced dose (33 or 25 microg) gonadotropin-releasing hormone agonist long protocol for multifollicular ovarian stimulation in patients with high basal serum follicle-stimulating hormone levels undergoing in vitro fertilization-embryo transfer. AB - The aim of the present study was to evaluate the clinical efficacy of half-dose (50 mug) and further reduced dose (33 or 25 mug) gonadotropin-releasing hormone agonist (GnRH-a; triptorelin) long protocols for multifollicular ovarian stimulation (MFOS) for patients with high basal serum follicle-stimulating hormone (FSH) level undergoing in vitro fertilization and embryo transfer (IVF ET). One hundred and two IVF-ET cycles performed in 84 infertile patients with high basal serum FSH (>10.0 mIU/ml) were included in this retrospective study. Study subjects were assigned to two groups: continuous half-dose GnRH-a long protocol (group A, n = 63) vs. further reduced dose GnRH-a long protocol (group B, n = 39) from half-dose at the start of GnRH-a to one-third or one-quarter dose after pituitary downregulation. Exogenous FSH or human menopausal gonadotropin was administered for MFOS in step-down mode, four or fewer embryos were transferred, and the outcomes of MFOS were compared between the two groups. Serum estradiol (E(2)) level on the day of human chorionic gonadotropin administration was significantly higher in group B (mean +/- standard deviation (SD): 1318.3 +/- 1120.4 vs. 2054.9 +/- 1773.5 pg/ml, p = 0.015). The number of transferable and good-quality embryos was also significantly higher in group B (mean +/- SD: 2.9 +/- 1.7 vs. 3.7 +/- 2.0, p = 0.027; 1.8 +/- 1.4 vs. 2.7 +/- 2.0, p = 0.020). No statistically significant difference in the outcomes was observed with respect to the dose of gonadotropins administered, the number of oocytes retrieved or the clinical pregnancy rate. In conclusion, GnRH-a long protocol with a reduced dose, tapered from the starting half-dose to a third or a quarter of the normal dose after pituitary suppression, may be beneficial for MFOS in IVF-ET patients with a high basal serum FSH level. A further prospective randomized controlled study on a larger scale is needed to confirm these findings. PMID- 16316846 TI - Controlled ovarian hyperstimulation: does prolonged stimulation justify cancellation of in vitro fertilization cycles? AB - BACKGROUND: In daily practice, assisted reproductive technology (ART) cycles are often cancelled under the assumption that a prolonged stimulation period lowers the likelihood of an appropriate ovarian response. The aim of the present study was to determine whether a prolonged cycle has an adverse effect on achievement of pregnancy. METHODS: The study sample included consecutive women enrolled in our ART unit between 1999 and 2001 who were treated with the mid-luteal long suppressive gonadotropin-releasing hormone protocol. Data were collected prospectively on a computerized database and evaluated at the end of the study. Prolonged stimulation was defined as a stimulation period of more than two standard deviations (SD) above the mean. Outcome was compared between patients who required prolonged stimulation and those who did not. RESULTS: A total of 1015 consecutive in vitro fertilization (IVF) cycles were performed with the mid luteal long suppressive protocol during the study period. Thirty-four women required prolonged stimulation. No difference in clinical pregnancy rate was detected between women who received prolonged stimulation and those who did not (9/34, 26.5% vs. 291/981, 29.7%), despite the significantly fewer oocytes retrieved in the prolonged-stimulation group (7.1 +/- 5.2 and 11.6 +/- 6.7 (mean +/- SD), respectively, p < 0.001). CONCLUSION: The likelihood of achieving pregnancy is not influenced by the length of stimulation. We recommend that IVF cycles should not be discontinued on the grounds of prolonged stimulation alone. PMID- 16316847 TI - Pathologic findings in hysteroscopy before in vitro fertilization-embryo transfer (IVF-ET). AB - BACKGROUND: The aim of this study was to evaluate hysteroscopy routinely performed prior to in vitro fertilization-embryo transfer (IVF-ET). METHODS: We analyzed in a prospective study 300 patients who underwent hysteroscopy before the first IVF-ET cycle. We analyzed then in a retrospective manner 300 patients who did not perform hysteroscopy. RESULTS: One-hundred-and-eighty (60%) hysteroscopies were normal but 120 (40%) revealed an unsuspected intrauterine abnormality. We did not find statistically significant differences between patients with normal or abnormal hysteroscopy in any characteristic. We found a statistically significant difference in pregnancy rate between women who performed hysteroscopy before IVF-ET cycle and in women who did not perform it. CONCLUSIONS: Hysteroscopy, as a routine examination, should be performed before the first IVF-ET cycle in all patients. PMID- 16316848 TI - Ovarian Leydig cell tumor in a peri-menopausal woman with severe hyperandrogenism and virilization. AB - The authors report a case of Leydig cell tumor in a 46-year-old woman who first presented with severe clinical hyperandrogenism and associated complex medical history. Investigations revealed markedly raised serum concentrations of testosterone (28.3 nmol/l) and free androgen index (54.4), whereas sex hormone binding globulin, random cortisol, androstenedione, 17-hydroxyprogesterone and dehydroepiandrosterone sulphate concentrations were all within the normal range. Transabdominal ultrasound and computed tomography scan of the pelvis and abdomen showed a slightly bulky right ovary, but no other abnormalities. An ovarian source of androgens was suspected and surgery was arranged. Following a three year history of defaulting appointments due to agoraphobia, she underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy and intraoperative selective ovarian venous sampling. Histopathological examination revealed a 2 cm Leydig cell tumor within the right ovary. Successful intraoperative ovarian venous sampling demonstrated significantly elevated testosterone levels (>260 nmol/l) from the right ovarian vein. Hyperandrogenaemia normalized post operatively. The patient showed significant regression of clinical signs and symptoms, including the anxiety disorder. Clinical presentation, biochemistry and imaging modalities should allow to detect androgen-secreting ovarian tumors, while selective venous sampling should be reserved for patients whom uncertainty remains. The present case confirms that androgen-secreting ovarian tumors represent a diagnostic and therapeutic challenge. They have to be considered in the differential diagnosis of severe hyperandrogenism even in peri-menopausal women. Although selective venous sampling is of diagnostic value, however, its impact on future management should be considered on individual basis. PMID- 16316849 TI - Compatibility studies between ibuprofen or ketoprofen with cellulose ether polymer mixtures using thermal analysis. AB - Differential scanning calorimetry (DSC) was used to investigate and detect incompatibilities between drugs such as: ibuprofen (IBU) or ketoprofen (KETO) with cellulose ether derivatives, which are frequently applied on controlled release dosage forms. Binary mixtures concerning methylcellulose (MC25) or hydroxypropylcellulose (HPC) with hydroxypropylmethylcellulose (HPMC) K15M or K100M in different ratios were prepared and evaluated by the appearance, shift, or disappearance of peaks and/or variations in the corresponding DeltaH values. According to the DSC results, binary mixtures between those polymers were found to be compatible, but their mixture with IBU or KETO, promotes a solid-solid interaction mainly with 1:1:1 (w/w) ratio (drug-excipient). However, when the drug:excipient interactions were detected, they were not found to affect the drug bioavailability. DSC was successfully employed to evaluate the compatibility of the drugs with the selected polymers. PMID- 16316850 TI - Press chamber coating as external lubrication for high speed rotary presses: lubricant spray rate optimization. AB - Lubrication of the tooling (punches and dies) is necessary to produce tablets. The most commonly used lubricant is magnesium stearate. Adding and blending magnesium stearate to the tablet mass often has negative effects on the properties of the compressed tablets (e.g., decreasing the tensile strength of the tablet). To avoid these negative effects, external lubrication systems were developed. This study investigated the functionality and the influence of a new press chamber coating system called the PKB II. The major difference between the PKB II and previous systems is its ability to spray a mixture of powdered magnesium stearate and air directly onto the punches and dies which was determined to allow the running of the rotor at higher speeds. The data showed a clear correlation between the spray rate of the lubricant and the concentration of the magnesium stearate per tablet. The PKB II was designed to allow for adjustments, in order to optimize the spray rate, by using the ejection force. The concentration of magnesium stearate was reduced to approximately 0.04% per tablet, using the PKB II. Additionally, the most common negative effects, such as the decrease in tablet tensile strength, were avoided by using this system. PMID- 16316851 TI - The hygroscopicity of moisture barrier film coatings. AB - The hygroscopicity of three commercial moisture-barrier film coatings, namely, Eudragit L30 D-55 (methacrylic acid-ethyl acrylate copolymer), Opadry AMB (polyvinyl alcohol based system), and Sepifilm LP 014 (hypromellose, microcrystalline cellulose, and stearic acid based formulation), was investigated using a dynamic vapor sorption apparatus. Moisture uptake by cast films and uncoated and coated tablet cores, which were designed to be hygroscopic, low hygroscopic, and waxy, was measured following exposure to repeat relative humidity (RH) cycles of 0-50-0-50-0%, 0-75-0-75-0%, and 0-90-0-90-0% RH at 25 degrees C. Eudragit cast film exhibited the fastest equilibration but was also the least hygroscopic. Sepifilm had the fastest sorption and took up the greatest mass of water. The rate of uptake for Opadry film was similar to Sepifilm. However, this film continued to sorb moisture for a longer period. When returned to 0% RH it retained moisture in the film showing that it had a high affinity for moisture within the film. The data for the different cores indicated that there was very little benefit in using a moisture barrier film on cores with low hygroscopicity, the mass gain being a sum of that which would be expected to sorb to the film and that which sorbs to the uncoated core. There was, however, some advantage for hygroscopic cores where, even though the barrier coatings allowed substantial water sorption into the core, the extent of this was less and the rate of uptake lower than for the uncoated sample. PMID- 16316852 TI - Preparation and evaluation of sustained ophthalmic gel of enoxacin. AB - The poor bioavailability and therapeutic response exhibited by conventional ophthalmic solutions due to rapid precorneal elimination of the drug may be overcome by the use of a gel system. The present work describes the formulation and evaluation of an ophthalmic delivery system containing an antibacterial agent, enoxacin, based on the concept of ophthalmic sustained gel, in which 2 hydroxypropyl-beta-cyclo-dextrin (HP-beta-CD) was used as a penetration enhancer in combination with hydroxypropylmethylcellulose (Methocel F4M) which acted as a vehicle. The developed formulation was therapeutically efficacious, nonirritant, and provided sustained release of the drug over 8 h period in vitro and 7 h period in vivo. The developed system is a viable alternative to conventional eye drops. PMID- 16316853 TI - Application of USP inlet extensions to the TSI impactor system 3306/3320 using HFA 227 based solution metered dose inhalers. AB - The objective of this study was to further evaluate the need for a vertical inlet extension when testing solution metered dose inhalers using the TSI Model 3306 Impactor Inlet in conjunction with the TSI Model 3320 Aerodynamic Particle Sizer (APS). The configurations tested using the TSI system were compared to baseline measurements that were performed using the Andersen Mark II 8-stage cascade impactor (ACI). Seven pressurized solution metered dose inhalers were tested using varied concentrations of beclomethasone dipropionate (BDP), ethanol, and HFA 227 propellant. The inhalers were tested with the cascade impactor, and with the TSI system. The TSI system had three different configurations as the manufacturer provided (0 cm) or with inlet extensions of 20 and 40 cm. The extensions were located between the USP inlet and the Model 3306 Impactor Inlet. There were no practical differences between each system for the stem, actuator, or USP inlet. The fine particle mass (aerodynamic mass < 4.7 microm) was affected by extension length and correlated well with the ACI when an extension was present. APS particle size measurements were unaffected by the extension lengths and correlated well to particle size determined from the ACI analysis. It has been confirmed that an inlet extension may be necessary for the TSI system in order to give mass results that correlate to the ACI, especially for formulations having significant concentrations of low volatility excipients. Additionally, the results generated from this study were used to evaluate the product performance of HFA 227 based solution formulations that contain varying concentrations of ethanol as a cosolvent. PMID- 16316854 TI - A new process for drug loaded nanocapsules preparation using a membrane contactor. AB - In this paper, we describe a new process for the preparation of drug loaded nanocapsules using a membrane contactor which may be scaled up for industrial applications. Nanocapsules are prepared according to the nanoprecipitation method. The organic phase (solvent, polymer, oil, and drug) is pressed through the pores of an ultrafiltration membrane via the filtrate side. The aqueous phase (water and surfactant) circulates inside the membrane module, and sweeps away the nanocaspules forming at the pore outlets. Two model drugs are selected for the preparation of drug loaded nanocapsules: indomethacin and vitamin E. It is shown that indomethacin loaded nanocapsules with a mean diameter of 240 nm and vitamin E loaded nanocapsules with a mean diameter of 230 nm are obtained with a 150,000 daltons ultrafiltration membrane, a transmembrane pressure of 3 bar, and a crossflow rate of 1.7 m.s(- 1). High fluxes are also obtained (around 0.6 m3/h.m2), leading to the preparation of 1.8 10(- 3) m3 drug loaded nanocapsules in 8 min. The advantage of this membrane contactor compared to other processes for drug loaded nanocapsules preparation is shown to be its scale-up ability. PMID- 16316855 TI - Comparison of three pharmaceutical products obtained from Mexico and the United States: a case study. AB - In recent years, there has been much debate concerning the relative pros and cons of purchasing medications from foreign markets such as Mexico and Canada. The following study compares the content uniformity and weight variation for three medicinal products, acquired from pharmacies in both Mexico and the United States: amoxicillin capsules (500 mg), amoxicillin/clavulanic acid suspension (400 mg and 57 mg/5 mL, respectively), and furosemide tablets (40 mg). Twenty capsules/tablets were individually weighed and a designated aliquot was taken. Following dissolution in an appropriate solvent and sonication, a sample was taken and analyzed via high performance liquid chromatography (HPLC). The suspensions were prepared according to directions on the label. Five samples of the suspensions were then taken and analyzed via an appropriate HPLC method. The content uniformity for the amoxicillin capsules was found to be 15.4 +/- 2.4% and 99.4 +/- 9.3%, for Mexican and U.S. capsules, respectively. The percent relative standard deviation (% RSD) for weight variation was found to be 8.7% and 1.5% for capsules obtained from Mexico and the United States, respectively. Content uniformity analysis for the Mexican suspension product resulted in an average of 85.5 +/- 1.2% for amoxicillin and 98.6 +/- 1.9% for the clavulanic acid content, while the results for the U.S. suspension product were 104.4 +/- 3.1% and 117.8 +/- 3.6% for amoxicillin and clavulanic acid, respectively. Content uniformity for the furosemide tablets was found to be 90.3 +/- 4.8% and 95.6 +/- 2.1% for Mexican and U.S. tablets, respectively. The % RSD of weight variation for the Mexican tablets was 2.1%, while the % RSD for the U.S. tablets was found to be 1.0%. From the three products tested, content analysis revealed that the amount of active ingredients for two of the products acquired in Mexico were appreciably less than the concentrations for their U.S. counterparts. PMID- 16316856 TI - Reversible Z-E isomerism and pharmaceutical implications for SU5416. AB - SU5416 (Z-isomer), the first in its class of angiogenesis inhibitors, in solution converts to the E-isomer following light exposure and reverts to the Z-isomer in the dark. Kinetics of this Z-E isomerism in pharmaceutical media is reported. Analytical solutions need light protection at 5 degrees C to maintain integrity. While E-isomer in light-exposed product increased to 0.9% in 24 hours, light protected product showed no change (25 degrees C, 18 months). Infusate studies indicated that < 1.9% E-isomer will be dosed to patients and would likely convert to the Z-isomer, following administration. This report implies Z-E isomerism in SU5416 is controllable with no limitations towards ensuring pharmaceutical product quality. PMID- 16316857 TI - Development and mathematical simulation of theophylline pulsatile release tablets. AB - Theophylline pulsatile release tablets consisting of a fast-swelling core with a water-insoluble ethylcellulose were developed. Effects of coating material, the amount of the plasticizer, subcoating, the type of the disintegrant, and coating level on the release profiles were investigated. Results showed that ethylcellulose was the best candidate polymer for pulsatile release tablets. Rupture time increased with increasing the amount of the plasticizer, but 15% plasticizer provided the best release profiles. Tablets with Methocel E50 as subcoating was most optimal in order to achieve a long lag time and followed by a rapid release. The lag time of tablets containing different disintegrants increased in the following order: croscarmellose (Ac-Di-Sol) < sodium starch glycolate (Explotab) < low-substituted hydroxypropyl cellulose (L-HPC) < crospovidone (Kollidon CL). And the rupture time increased with higher coating level. A mathematical model was presented to predict the lag time prior to rupture. Results of the water uptake experiment were used to estimate the apparent diffusion coefficient of the coating tablets. The prediction of the lag time based on the presented model is in good agreement with the experimental results. PMID- 16316858 TI - Artificial neural networks: comparison of two programs for modeling a process of nanoparticle preparation. AB - Artificial Neural Networks (ANNs) were used to predict nanoparticle size and micropore surface area of polylactic acid nanoparticles, prepared by a double emulsion method. Different batches were prepared while varying polymer and surfactant concentration, as well as homogenization pressure. Two commercial ANNs programs were evaluated: Neuroshell Predictor, a black-box software adopting both neural and genetic strategies, and Neurosolutions, allowing a step-by-step building of the network. Results were compared to those obtained by statistical method. Predictions from ANNs were more accurate than those calculated using non linear regression. Neuroshell Predictor allowed quantification of the relative importance of the inputs. Furthermore, by varying the network topology and parameters using Neurosolutions, it was possible to obtain output values which were closer to experimental values. Therefore, ANNs represent a promising tool for the analysis of processes involving preparation of polymeric carriers and for prediction of their physical properties. PMID- 16316859 TI - Bioequivalence evaluation of two formulations of doxazosin tablet in healthy thai male volunteers. AB - The bioequivalence of two doxazosin 2 mg tablets was determined in 24 healthy Thai male volunteers after one single dose in a randomized cross-over study with a one week washout period. The study was conducted at Faculty of Pharmaceutical Sciences and Health Sciences Research Institute, Naresuan University, Phitsanulok, Thailand. Reference (Cardura, Heinrich Mack Nachf. GmbH & Co. GK, Illertissen, Germany) and test (Dozozin-2, Umeda Co., Ltd., Bangkok Thailand) were administered to volunteers after overnight fasting. Blood samples were collected at specified time intervals and plasma was separated. The validated HPLC method with fluorescence detection was used for quantification of doxazosin in plasma samples. The pharmacokinetic parameters, T(max), C(max), AUC(t), AUC(infinity), T(1/2), lambda(z), Cl and V(d), were determined from plasma concentration time profile of both formulations by using non-compartment analysis. The calculated pharmacokinetic parameters were compared statistically to evaluate bioequivalence between the two brands. The analysis of variance (ANOVA) using log-transformed C(max), AUC(t), and AUC(infinity) did not show any significant difference between two formulations. The point estimates and 90% confidence intervals for C(max), AUC(t) and AUC(infinity) were within the acceptance range (0.80-1.25), satisfying the bioequivalence criteria of the Thailand Food and Drug Administration Guidelines. These results indicate that Dozozin-2 is bioequivalent to Cardura and, thus, may be prescribed interchangeably. PMID- 16316860 TI - Topochemical models for prediction of anti-HIV activity of 4-benzyl pyridinone derivatives. AB - Relationship between topochemical indices and anti-HIV activity of 4-Benzyl pyridinone derivatives has been investigated. The values of molecular connectivity topochemical index (an adjacency based topochemical descriptor) Wiener's topochemical index (a distance based topochemical descriptor) and superadjacency topochemical index (an adjacency cum distance based topochemical descriptor) were calculated for each of the 32 compounds comprising the data set using an in-house computer program. The resultant data was analyzed and suitable models were developed after identification of the active ranges. Subsequently, a biological activity was assigned using these models to each of the compounds involved in the dataset which was then compared with the reported anti-HIV activity. Exceptionally high accuracy of prediction was observed using these models. These models offer vast potential for providing lead structures for the development of potent anti-HIV agents. PMID- 16316864 TI - Combination inhalers containing inhaled corticosteroids and long-acting beta2 agonists: improved clinical efficacy and dosing options in patients with asthma. AB - Combination therapy with inhaled corticosteroids (ICS) and long-acting beta2 agonists (LABA) is a recognized treatment for adults with moderate to severe asthma. The introduction of inhalers containing both an ICS and a LABA simplifies treatment and improves asthma control. This review discusses clinical evidence that budesonide/formoterol and salmeterol/fluticasone are effective and well tolerated in asthma treatment. Moreover, the rapid onset of effect and long duration of action of budesonide and formoterol make once-daily dosing, adjustable maintenance dosing, and the novel treatment strategy of using budesonide/formoterol for maintenance and as needed for symptom relief, valuable treatment options for patients with asthma. PMID- 16316865 TI - The relationship between specific anxiety syndromes and somatic symptoms in adolescents with asthma and other chronic diseases. AB - BACKGROUND: The impact of a chronic disease on the emotional well-being of children and adolescents is controversial in the literature. This study tested the hypotheses that 1) a specific approach is required to assess emotional deviations in adolescents with chronic diseases and 2) specific anxiety symptoms are predictive of excessive somatic symptoms. METHODS: Emotional and somatic symptoms were measured in four groups, selected from a community sample of 897 adolescents: 32 with asthma, 20 with other severe chronic diseases, 30 with median scores (the true comparison group), and 29 with minimal scores on common measures of trait anxiety and depression. RESULTS: The asthma and chronic disease groups scored not significantly higher than the true comparison group on trait anxiety, depression, negative affectivity, five anxiety syndromes, anxiety related physical, and miscellaneous somatic symptoms. The asthma and chronic disease groups scored only higher than the true comparison group on panic attacks and respiration symptoms. Regression analyses showed that severity of asthma was no significant factor, and the minimal group scored consistently lower than the other groups, except on physical injury fears. There were no group differences in positive affect. Girls scored higher than boys on specific anxiety syndromes (except on obsessive-compulsive disorder) and also on respiration symptoms. CONCLUSION: Adolescents with severe chronic diseases deviated from a true comparison control group on panic attacks, but not on other negative and positive emotions. PMID- 16316866 TI - Neighborhood racial composition and availability of asthma drugs in retail pharmacies. AB - This study investigated whether there may be differences in the availability of asthma drugs and equipment in retail pharmacies in nonwhite and white neighborhoods in the District of Columbia. We conducted a telephone survey of a random sample of 38 retail pharmacies in predominately black or white neighborhoods. No differences in the reported availability of asthma drugs and equipment by neighborhood were found, although there was variability in availability of certain asthma drugs and limited availability of asthma equipment. Low demand was the most frequent reason cited by pharmacists regarding why asthma drugs or equipment were not in stock. PMID- 16316867 TI - A comparison of virus-associated and multi-trigger wheeze in school children. AB - To examine differences between virus-associated wheeze and wheeze associated with other triggers (multi-trigger wheeze) in elementary school children, we performed a cross-sectional school-based questionnaire study of 5,998 children mainly 7 to 12 years of age, with outliers 6 and 13 years of age. Using parent-completed questionnaires, we identified 522 children who wheezed only during upper respiratory tract infections (virus-associated wheeze), 1,186 children who wheezed on other occasions (multi-trigger wheeze), and 4,290 children with no wheeze. In comparison with children who had multi-trigger wheeze, children with virus-associated wheeze were more likely to be male, to be younger, and to have less frequent wheezy episodes. They were less likely to have night cough, shortness of breath or chest tightness, to have a personal or parental history of atopic disorders, to have a diagnosis of asthma, or to be receiving asthma treatment. Both types of wheeze were associated with social deprivation, a relationship that persisted after controlling for family smoking. Virus associated wheeze is a common but diminishing problem in this age group, and the differences between virus-associated wheeze and multi-trigger wheeze already noted in pre-school children persist in this older age group. PMID- 16316868 TI - Fever, use of antibiotics, and acute gastroenteritis during infancy as risk factors for the development of asthma in Korean school-age children. AB - OBJECTIVE: The hygiene hypothesis postulates that infections early in life might influence the development of asthma later in childhood. However, this hypothesis is controversial. The objective of this study was to determine whether there is a relationship between infection-related risk factors during infancy and development of asthma later in childhood. METHODS: Data were obtained by a nationwide, population-based, cross-sectional study of 26,400 children, 7 to 12 years of age. Parents completed a questionnaire on symptoms of and risk factors for asthma, including the number of fever episodes, acute gastroenteritis, use of antibiotics, and immunization during infancy. The presence of asthma was defined by parental report of "wheeze in the last 12 months" or "asthma ever." Data were analyzed by logistic regression analysis. RESULTS: The occurrence of fever during infancy was weakly associated with the development of asthma (P < 0.05). A higher frequency of episodes of fever was associated with a higher risk of development of asthma. The relationship between antibiotic use during infancy and development of asthma was also significant (P < 0.0001). The adjusted odds ratios of childhood asthma increased in proportion to the number of antibiotic courses during infancy. A history of acute gastroenteritis during infancy increased the risk of developing asthma in later life (P < 0.001). In contrast, immunization in infancy was not related to development of asthma in childhood (P > 0.05). CONCLUSIONS: These data suggest that the development of childhood asthma is associated with episodes of fever, antibiotic use, and acute gastroenteritis during infancy. PMID- 16316869 TI - Perceived control and quality of life in asthma: impact of asthma education. AB - The purpose of this study was to determine the relationship between patients' perception of asthma control and generic and asthma-specific quality of life (QOL) post-completion of a behavior modification-based adult asthma education program. A secondary objective was to examine associations between changes in perceived control of asthma and generic and asthma-specific QOL. Outcome measures were collected via an asthma management questionnaire (AMQ), generic (SF-36) and asthma-specific (AQLQ) QOL questionnaires, and a perceived control of asthma questionnaire (PCAQ). The cohort (n = 55) consisted of predominately female (75%), married (56%), middle income (46%) patients with severe asthma (65%) who had completed a university or college education (20%) and were working full-time (42%). The mean age was 45.2 (SD = 17.5) years. Perceived control of asthma and generic and asthma-specific quality of life significantly improved after completing the behavior modification-based adult asthma education program. Significant associations were found between perceived control of asthma (PCAQ) and both generic (SF-36) and asthma-specific QOL (AQLQ). Baseline PCAQ was related to all four domains and the total score of the AQLQ and 5 of the 8 domains of the SF-36. PCAQ was related to 3 of the 4 AQLQ domains at 3 months and total AQLQ score at both 1 and 3 months post-education. PCAQ was related to all 8 domains of the SF-36 at 1 month; and 4 of 8 domains at 3 months. Change in PCAQ (deltaPCAQ) was related to change in symptom score, emotional functioning, and total AQLQ score from baseline to 1 month and change in symptom score from baseline to 3 months. In conclusion, perceived control of asthma in patients participating in a behavior modification-based asthma education program was related to generic and disease-specific QOL. An improvement in PCAQ was associated with improved QOL following asthma education. Using the PCAQ as part of an asthma educational needs assessment may be a quick, simple way to identify and target education towards asthma patients with low perceived control. PMID- 16316870 TI - A multiple cause-of-death analysis of asthma mortality in the United States, 1990 2001. AB - OBJECTIVES: Most analyses of asthma mortality in the United States have relied solely on underlying cause-of-death data, which may underestimate the magnitude of asthma-related mortality. We used multiple cause-of-death data to examine asthma-related mortality trends in the United States. METHODS: Data were selected from the United States Multiple Cause-of-Death Files, 1990-2001. Mortality rates and 95% confidence intervals were computed to examine differences in asthma mortality over time and by age, race/ethnicity, and gender. Location of death and seasonal variations in asthma mortality were also assessed, as well as the impact of seasonal respiratory infections. RESULTS: We identified 135,668 asthma-related deaths in the United States over the 12-year period, representing an age-adjusted mortality rate of 4.4 per 100,000. Only 45% of the asthma-related deaths had asthma recorded as the underlying cause. Whites and older adults were less likely to have asthma listed as the underlying cause. Asthma mortality rates mirrored underlying cause trends, increasing slightly between 1990 and 1995, declining between 1996 and 1998, and further declining after International Classification of Disease (ICD)-10 implementation in 1999. Mortality was highest among blacks and the elderly and was higher among females than males. Asthma-related deaths peaked in the winter months and were over four times more likely than non-asthma deaths to have acute upper respiratory infections, influenza, or acute bronchitis listed on the death record. The proportion of asthma-related deaths occurring outside a medical setting increased steadily over the period, from 23.3% in 1990 to 29.4% in 2001. CONCLUSIONS: The burden of asthma may be underestimated by relying solely on underlying cause-of-death data. Further research is needed to determine the reasons for the steady increase in out-of-hospital deaths and the continued demographic disparities in mortality. PMID- 16316871 TI - Does specific immunotherapy injection cause an increase in bronchial reactivity? AB - BACKGROUND: Several well-controlled studies have proven the clinical benefit of specific immunotherapy (SIT) for seasonal allergic rhinitis (AR). However, whether subcutaneous SIT injection could cause a transient increase in bronchial reactivity (BR) remains unknown. OBJECTIVE: To investigate whether subcutaneous SIT injection, either during or outside the pollen season, could cause an increase in BR in children with pollen allergy. METHODS: Twenty-two children (mean age 13.6 +/- 0.7 years) with AR who were receiving maintenance SIT for 15 months were included in the study. Pre-injection BR of the patients was evaluated with methacholine provocation test immediately before maintenance dose of SIT during the peak pollen season and outside the season. The post-injection test was administered 24 hours after SIT injection. RESULTS: There was no difference in FEV1 measures recorded during [98(93-109)%] and outside [102(96-111)%] the pollen season. There was no significant difference between pre- [64(7-64) mg/mL] and post-allergen injection [32(7.5-64) mg/mL] BR outside the pollen season (p = 0.9). A trend towards improvement following allergen injection [64(5.4-64)] as compared to pre-allergen injection [14.6(3.5-64)] was shown during the pollen season (p = 0.053). Although PC20 measures in the pollen season were lower than outside the season, the difference was not significant. The percentage of the patients with bronchial hyperreactivity was 62% during and 43% outside the season. CONCLUSION: SIT injections both during and outside the pollen season cause no increase in BR in children with AR. This calls into question the necessity of empirical dose reduction during the pollen season. PMID- 16316872 TI - Patient satisfaction with budesonide Turbuhaler versus triamcinolone acetonide administered via pressurized metered-dose inhaler in a managed care setting. AB - Dissatisfaction with medication may negatively affect compliance and thus the effectiveness of the treatment. However, no prospective well-controlled studies have assessed the relative patient satisfaction with competing inhaled corticosteroids in a real-life setting. The objective of the current study was to compare the relative patient satisfaction with budesonide inhalation powder administered via Turbuhaler (AstraZeneca LP, Wilmington, DE) (200 to 1600 microg/d using one of 3 dosing strengths: 100, 200, or 400 microg per inhalation) and triamcinolone acetonide administered via pressurized metered-dose inhaler (200 to 1600 microg/d) among persons treated in managed care settings. A total of 945 subjects 18 years of age or older diagnosed with asthma and enrolled in 25 managed care organizations participated in this prospective, randomized, open label, parallel-group, 12-month study. As part of the study, subjects completed a self-administered, 17-item patient satisfaction questionnaire that addressed 4 domains: side effects, knowledge/ease of use, convenience, and overall satisfaction. Questionnaire reliability was assessed using Cronbach's alpha, and validity was examined by correlating subscale scores with symptom-free days and Medical Outcomes Study 36-Item Short-Form questionnaire and Asthma Quality of Life Questionnaire scores. The satisfaction questionnaire also included a previously validated section addressing patient compliance. Patients receiving budesonide had significantly higher scores for all four satisfaction subscales throughout the study period than did those receiving triamcinolone acetonide. Similarly, compliance scores were consistently higher for the budesonide group. The difference between the treatment groups in overall satisfaction scores at the end of the study was clinically meaningful. Patients treated with budesonide were significantly more satisfied and compliant with their inhaled corticosteroid regimen compared with patients treated with triamcinolone acetonide. PMID- 16316873 TI - Sex differences in asthma prevalence and other disease characteristics in eight states. AB - OBJECTIVES: We assessed the sex differences in asthma prevalence and asthma control characteristics within eight states. METHODS: We analyzed data from the 2001 Behavioral Risk Factor Surveillance System survey. RESULTS: Lifetime and current asthma prevalence were higher for females in each of the eight states compared to males. Adult onset of asthma was reported more often by females with current asthma, and childhood onset was reported more often by males. Sex differences were identified for the eight asthma-control characteristics. CONCLUSIONS: Females in eight states presented higher asthma risk and poorer asthma profiles than males. State surveillance data can be used to identify disparities and to develop appropriate public health interventions. PMID- 16316874 TI - Relationship between peak expiratory flow rate and shoulders posture in healthy individuals and moderate to severe asthmatic patients. AB - INTRODUCTION: Asthma overloads respiratory muscles causing changes in posture. OBJECTIVES: To evaluate shoulder position in asthmatics and its correlation to the peak expiratory flow rate (PEFR). METHODS: A cross-sectional study involving 19 asthmatics (study group) and 20 healthy volunteers (control group). Assessment of PEFR was used to determine airway obstruction. Shoulder position was quantified as the ratio between the front and back measurements, in centimeters, of the distance between the two outermost points of each scapula. Shoulder position measurement (SPM) ratio data from both groups were compared and correlated to PEFR. RESULTS: The asthmatics presented lower PEFR (279 +/- 64 L/min) and SPM (0.88: range 0.83-0.9) than did control subjects (555 +/- 100 L/min and 0.97: range 0.95-1, respectively) (p < 0.001). There was a correlation between PEFR and SPM (asthmatics r = 0.8; controls r = 0.5). CONCLUSION: Determining SPM ratios allowed us to identify those postural alterations that correlated with pulmonary function in asthmatics and in healthy individuals. Further investigation is needed before SPM can be used as a tool for clinical evaluation of asthma patients. PMID- 16316875 TI - Asthma among rural Minnesota adolescents. AB - This study examined the prevalence of and risk factors for wheezing and asthma in rural Minnesota adolescents. A survey querying about wheezing, asthma, farm residence, and other respiratory-related factors was administered to all 9th to 11th grade students (N = 13,490) in a stratified, random sample of high schools in rural Minnesota. Nearly one in 8 (12.6%) students reported ever-diagnosed asthma, and 9.2% reported current asthma. Students living on farms reported less wheezing and asthma than rural non-farm students. Obesity and smoking were associated with wheezing and asthma in all rural adolescents. Asthma rates increased with age among girls and may be largely accounted for by simultaneous increases in smoking rates. Despite the apparent protective effect of farm residence, asthma remains a significant public health issue among rural Minnesota adolescents. PMID- 16316876 TI - Evaluation of factors affecting peak expiratory flow in healthy adults: is it necessary to stand up? AB - BACKGROUND: Guidelines for the Diagnoses and Management of Asthma recommend that the peak flow meter should be used in a standing position. OBJECTIVE: To determine whether the peak expiratory flow (PEF) varies with the position of the subject and to evaluate the impact of application of nose clip on PEF measurement. METHODS: Using a Mini Wright Peak Flow Meter, the PEF was measured in 33 healthy adults in sitting and standing positions, with and without application of a nose clip. Each subject blew into the peak flow meter three times and best of the 3 blows was recorded. RESULTS: The PEF was not significantly higher in standing than in sitting position. Application of nose clip had no significant impact on the PEF. CONCLUSION: The position of the subject and application of nose clip has no significant impact on PEF measurement. PMID- 16316877 TI - The validity and reliability of two asthma knowledge questionnaires. AB - BACKGROUND: This study aimed to develop and validate two asthma knowledge questionnaires, one for consumers (CQ) and one for health care professionals (HQ) to fill an existing gap in the literature. METHODS: Following development and pilot testing, the revised CQ and HQ were administered to respiratory physicians, pharmacists and people with asthma and without asthma. RESULTS: Both the CQ and HQ were shown to be reliable with Cronbach's alpha of 0.78 and 0.92 respectively. Both were also found to have good content, face, construct and discriminant validity. CONCLUSION: The CQ and HQ were shown to be simple, valid and reliable instruments for the assessment of asthma knowledge of consumers and health care professionals and to direct educational resources more appropriately. Future research will evaluate the utility of these instruments in clinical practice. PMID- 16316879 TI - Editor's preamble: the nature of IBS and the need for a psychological approach. PMID- 16316880 TI - Hypnosis for irritable bowel syndrome: the empirical evidence of therapeutic effects. AB - Irritable bowel syndrome (IBS) is a complex and prevalent functional gastrointestinal disorder that is treated with limited effectiveness by standard medical care. Hypnosis treatment is, along with cognitive-behavioral therapy, the psychological therapy best researched as an intervention for IBS. Eleven studies, including 5 controlled studies, have assessed the therapeutic effects of hypnosis for IBS. Although this literature has significant limitations, such as small sample sizes and lack of parallel comparisons with other treatments, this body of research consistently shows hypnosis to have a substantial therapeutic impact on IBS, even for patients unresponsive to standard medical interventions. The median response rate to hypnosis treatment is 87%, bowel symptoms can generally be expected to improve by about half, psychological symptoms and life functioning improve after treatment, and therapeutic gains are well maintained for most patients for years after the end of treatment. PMID- 16316881 TI - Effective management of irritable bowel syndrome--the Manchester Model. AB - Over the years, researchers have shown that hypnotherapy can be exceptionally helpful in the management of refractory irritable bowel syndrome. However, it is a labor-intensive modality with a finite success rate and is not suitable for everyone. It is therefore best incorporated into a program of graduated care that has a contingency plan for dealing with individuals who do not respond to this particular form of treatment. This paper describes how hypnotherapy has been successfully integrated into the functional gastroenterology service in Manchester. PMID- 16316882 TI - Gut-directed hypnotherapy: the Manchester approach for treatment of irritable bowel syndrome. AB - This article describes the particular approach of using hypnosis as an adjunct to treating irritable bowel syndrome, developed within the Department of Medicine at the University Hospital of South Manchester, UK, since the 1980s. Patients receive up to 12 sessions over a 3-month period, and the majority of patients achieve marked improvement in symptoms and quality of life, an effect that is usually sustained. The therapy has a "gut-directed" framework that aims to teach patients the necessary hypnotic skills to control gut function and reduce symptoms, such as hand warmth on the abdomen and imagery. Other interventions based on particular lifestyle and psychological factors commonly found to influence symptoms are also included as appropriate for the individual patient. PMID- 16316883 TI - Standardized hypnosis treatment for irritable bowel syndrome: the North Carolina protocol. AB - The North Carolina protocol is a seven-session hypnosis-treatment approach for irritable bowel syndrome that is unique in that the entire course of treatment is designed for verbatim delivery. The protocol has been tested in two published research studies and found to benefit more than 80% of patients. This article describes the development, content, and testing of the protocol, and how it is used in clinical practice. PMID- 16316884 TI - Hypnosis for irritable bowel syndrome: the quest for the mechanism of action. AB - Irritable bowel syndrome (IBS) is a very common condition in the Western part of the world, and it accounts for a large proportion of the workload of a gastroenterologist. Unfortunately, the pathogenesis and pathophysiology of the syndrome are incompletely understood, and the treatment options are limited. However, hypnotherapy is one treatment option that has proven to be very useful in IBS. The mechanisms of action explaining why hypnosis is effective for IBS are not altogether known, but recent studies have shed some light on this issue. These studies, and what can be learned from them about how hypnosis impacts IBS, are reviewed in this article. Hypnosis may affect IBS partly through changes in colorectal sensitivity and improvement in psychological factors. The effects on GI motility and the autonomic nervous system are less clear and need further evaluation. PMID- 16316885 TI - Hypnosis home treatment for irritable bowel syndrome: a pilot study. AB - Hypnosis treatment often improves irritable bowel syndrome (IBS), but the costs and reliance on specialized therapists limit its availability. A 3-month home treatment version of a scripted hypnosis protocol previously shown to improve all central IBS symptoms was completed by 19 IBS patients. Outcomes were compared to those of 57 matched IBS patients from a separate study receiving only standard medical care. Ten of the hypnosis subjects (53%) responded to treatment by 3 month follow-up (response defined as more than 50% reduction in IBS severity) vs. 15 (26%) of controls. Hypnosis subjects improved more in quality of life scores compared to controls. Anxiety predicted poor treatment response. Hypnosis responders remained improved at 6-month follow-up. Although response rate was lower than previously observed in therapist-delivered treatment, hypnosis home treatment may double the proportion of IBS patients improving significantly across 6 months. PMID- 16316886 TI - Effects of tailored and manualized hypnotic inductions for complicated irritable bowel syndrome patients. AB - This modest clinical pilot study was intended to provide preliminary data on the effects of hypnotic inductions tailored to an irritable bowel syndrome patient at each session compared to Palsson's manualized protocol. Patients (N = 8) who had not previously responded to any form of treatment were assigned randomly to either a tailored or manualized induction condition. Other than pretesting for hypnotizability, the procedure followed for the manualized group (n = 4) was exactly as prescribed by O. Palsson (1998). The identical procedure was used for the other 4 patients except that the inductions were individualized. All 8 patients showed favorable responses to treatment immediately posttreatment and at 10-month follow-up. Only the tailored group showed no incapacitating pain at posttreatment but greater emotional stress than the manualized group. The tailored group continued to improve and showed better results than the manualized group at 10 months, and the posttreatment emotional distress was significantly attenuated. PMID- 16316887 TI - BASES (British Association of Sport and Exercise Sciences) Annual Conference, 4-7 September 2005, Loughborough University, UK. Abstracts. PMID- 16316890 TI - Therapeutic efficacy of infliximab on active Crohn's disease under nutritional therapy. AB - OBJECTIVE: The aim of this investigation was to elucidate retrospectively the therapeutic effect of infliximab in patients with active Crohn's disease (CD) under nutritional therapy. MATERIAL AND METHODS: Using a review of the clinical records in 24 nationwide institutions specializing in inflammatory bowel disease, the short-term effect of infliximab in 97 patients with active CD was retrospectively investigated. The Crohn's disease activity index (CDAI) at baseline and after 2 weeks of a single infliximab administration (5 mg/kg) was compared among patients under total parenteral nutrition (TPN group, n=36), those following an elemental or polymeric diet (EN group, n=49) and those without TPN and EN (NN group, n=12). A decrease in CDAI >or= 70 or a CDAI value <150 at 2 weeks was regarded as effective. RESULTS: There was no difference in CDAI at baseline among the three groups. In each group, CDAI decreased significantly (from 250 (195-290) [median (interquartiles)] to 152 (123-233) in the TPN group, p<0.0001; from 259 (200-325) to 180 (130-238) in the EN group, p<0.0001; from 278 (222-291) to 164 (132-196) in the NN group, p=0.003). Infliximab was effective in 63.9% of patients in the TPN group, in 55.1% of those in the EN group and in 75% of the NN group. There was no statistical difference in efficacy among the three groups (p=0.4). Multivariate logistic regression analysis revealed younger age to be a significant factor related to the efficacy of infliximab. CONCLUSIONS: Infliximab is effective in patients with CD under TPN or EN. Age at infliximab administration may be predictive of response to infliximab. PMID- 16316891 TI - Preventive effect of nutritional therapy against postoperative recurrence of Crohn disease, with reference to findings determined by intra-operative enteroscopy. AB - OBJECTIVE: The aim of this study was to elucidate the predictive value of intra operative enteroscopy (IOE) and the effect of enteral nutrition (EN) with regard to the postoperative recurrence of Crohn disease (CD). MATERIAL AND METHODS: Forty patients requiring surgery for severe intestinal complications of CD were examined by IOE, and the severity of the remnant small intestine was determined. Patients were subclassified into either an EN group (>1,200 kcal/day) or a non-EN group (<1,200 kcal/day) according to the amount of daily EN intake after surgery. Contributions of IOE findings and EN to postoperative recurrence were analysed retrospectively. RESULTS: IOE identified intestinal lesions in 39 patients and active intestinal lesions in 24 patients. The cumulative rate of postoperative recurrence was significantly higher in patients with cobblestone appearance confirmed by IOE (p=0.006). However, other active intestinal lesions were not related to postoperative recurrence. EN reduced the cumulative rate of postoperative recurrence (p=0.017), especially in patients with penetrating type (p=0.005), in patients who did not have colitis (p=0.051) and in patients who did not have active intestinal lesions confirmed by IOE (p=0.02). CONCLUSIONS: EN is a prophylactic that prevents the postoperative recurrence of small intestinal CD. Patients with the penetrating type of CD, and those who do not have active lesions in the small intestine according to IOE, are candidates for EN after surgery. PMID- 16316892 TI - -137 (G/C) IL-18 promoter polymorphism in patients with inflammatory bowel disease. AB - OBJECTIVE: There is strong evidence that genetic factors contribute to the susceptibility for inflammatory bowel diseases (IBD). Recently, IL-18 promoter polymorphisms were characterized as risk factors for inflammatory diseases such as sepsis, asthma and adult-onset Still's disease. The aim of this study was to determine whether the -137 (G/C) IL-18 promoter polymorphism was associated with IBD susceptibility. MATERIAL AND METHODS: For association analysis, 470 patients with Crohn's disease (CD), 235 unrelated patients with ulcerative colitis (UC) and 347 controls were enrolled. Furthermore, 233 UC and 470 CD trios were included for segregation analysis. Genotyping was performed by application of the TaqMan MGB biallelic discrimination system. RESULTS: When comparing genotype frequencies of CD and UC patients versus controls, no significant difference was found (p=0.089 and p=0.078, respectively). However, the Cochran-Armitage trend test revealed a rising probability for CD and UC with increasing number of G alleles (p=0.030 and 0.028, respectively) for the case-control analysis. On the contrary, the family-based transmission disequilibrium test (TDT) did not show an association of the G allele with CD or UC in 470 CD and 233 UC trios (p=0.53 and p=0.79, respectively). CONCLUSION: The -137 (G/C) IL-18 promoter polymorphism is not a susceptibility factor for IBD in a German cohort. PMID- 16316893 TI - Saliva Interleukin-6 in patients with inflammatory bowel disease. AB - OBJECTIVE: The inflammatory bowel diseases (IBD), Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory conditions that affect the gastrointestinal tract. In regulation of this inflammatory process, Interleukin-6 (IL-6) has a major role. Overproduction of IL-6 by immunocompetent cells contributes to development of the inflammatory condition. Elevated levels of IL-6 in saliva could be expected, because the saliva-producing cells are part of the digestive system. MATERIAL AND METHODS: IL-6 concentrations in saliva and plasma were studied in patients with CD (n=15), UC (n=7) and reference persons (RP) (n=19) by use of an ELISA method. RESULTS: A significant difference in saliva IL 6 concentration between CD patients (median 16.9 ng/L; p<0.05) and RP (median 6.3 ng/L) was found. A significant difference in plasma IL-6 concentration between CD (median 10.3 ng/L; p<0.001) or UC (median 7.8 ng/L; p<0.001) and RP (median 0.8 ng/L) was observed. In patients with CD, plasma IL-6 correlated significantly with C-reactive protein (CRP) as well as albumin. In patients with UC, saliva IL 6 and plasma IL-6 correlated significantly with AI (activity index) scores as well as albumin. In patients with UC, a significant correlation between the saliva and plasma IL-6 concentrations was found. CONCLUSIONS: IL-6 was found in saliva in patients with IBD, documenting the general involvement of the gastrointestinal tract extending to the mouth cavity, and measuring IL-6 may be an additional method for evaluating and monitoring the disease activity. PMID- 16316894 TI - Incidence of colorectal cancer in a population-based cohort of patients with Barrett's oesophagus. AB - OBJECTIVE: Previous studies have shown a positive association between colorectal cancer and Barrett's oesophagus, but this association is disputed. No population based studies have examined the incidence of this cancer in patients with Barrett's oesophagus. MATERIAL AND METHODS: The present study comprised a population-based cohort of patients with Barrett's oesophagus (constructed using pathology reports of all oesophageal biopsies in Northern Ireland 1993-99; cohort subclassified according to whether specialized intestinal metaplasia (SIM) was present, absent, or not commented on in biopsies). Cases of colorectal cancer were identified by linking with the Northern Ireland Cancer Registry. The comparison group used was the general population in Northern Ireland. RESULTS: A total of 2969 patients with Barrett's oesophagus were followed for a total of 14,014 person-years (mean 4.7 years). SIM was present in 1670 patients (56.2%), absent in 545 (18.4%) and not commented on in 754 (25.4%). Colorectal cancer was diagnosed in 39 patients; 22 patients had cancer diagnosed at least 6 months after diagnosis of Barrett's oesophagus. There was no increased risk of colorectal cancer: the standardized incidence ratio (SIR) for cancer diagnosed at least 6 months after entry into the cohort was 0.82 (95% CI, 0.48-1.17); this risk did not alter with SIM status or gender. To assess a possible effect of diagnostic bias, we calculated SIRs for cancers occurring after at least 3 months, after at least 1 month and at any time after diagnosis of Barrett's oesophagus. These were 0.94 (0.57-1.30), 1.09 (0.69-1.48) and 1.46 (1.00-1.92), respectively. CONCLUSIONS: The incidence of colorectal cancer was not elevated in patients with Barrett's oesophagus. Diagnostic bias may explain why previous studies have found an association. PMID- 16316895 TI - Callous-unemotional traits are associated with clinical severity in referred boys with conduct problems. AB - Clinical practice with the heterogeneous group of children that present with conduct problems may benefit from improved formats for diagnostic subtyping. We investigated whether high levels of callous-unemotional traits (i.e. lack of empathy, remorselessness and shallow affects) would differentiate clinic-referred conduct-problem boys from those low on such traits. A consecutive series of 41 boys with conduct problems (6-13 years, mean age=9.60 years) referred to public child psychiatric units in Sweden were studied with data elicited from caregivers. Conduct-problem boys with many callous-unemotional traits had significantly more pervasive, varied and aggressive disruptive behavioural problems than boys low on these traits had. Higher levels of conduct problems in subjects with callous-unemotional traits were not explained by confounding presence of DSM-IV attention-deficit hyperactivity disorder and oppositional defiant disorder/conduct disorder symptoms. Boys with callous-unemotional traits also experienced poorer household circumstances and lived in families under high stress. Interestingly, they less often received help in school from special teachers but were more often diagnosed with dysthymia than boys low on callous unemotional traits. We conclude that callous-unemotional traits might designate a subgroup of boys with different aetiology to their conduct problems and possibly with specific treatment needs. However, the findings need to be replicated with larger samples. PMID- 16316896 TI - Significant others of persons with mental health problems: the testing of a questionnaire on the burden of significant others. AB - The importance of family and friends for patients with mental health problems has been recognized over the past few years. Significant others (SO) of patients admitted to a psychiatric ward specializing in affective disorders and suicide prevention filled in a self-rating questionnaire concerning their burden as SOs, the Involvement Evaluation Questionnaire (IEQ). The aims of this study were twofold: first, to test whether the questionnaire IEQ was applicable in psychiatric services in Sweden and second, to relate the IEQ results to socio demographic data and diagnosis of the patients, and also to the situation of the SOs. We found the questionnaire useful. Nearly half of the SOs had difficulties in accepting and handling the patient's psychiatric problems. More than half had been worried that the patient was going to harm him/herself and because of this had kept close watch on the patient. Almost all SOs worried about the patient's general health and future. Most of them felt obliged to intervene in the patient's activities of daily living. More than half worried about the kind of medical and psychological treatment the patient was receiving. Most SOs concluded that they were burdened by their engagement in the patient. Our results were compared with a European study of schizophrenic patients also investigated with the IEQ. The comparison indicated that the perceived burden on SOs of these different kinds of psychiatric patients can be equally burdensome. SOs need information, help and support in their difficult support role. This questionnaire could be useful in finding new routines for increasing the involvement by SOs in the treatment of patients with mental health problems. PMID- 16316897 TI - Using the Karolinska Scales of Personality on male juvenile delinquents: relationships between scales and factor structure. AB - The aim of the present study was to investigate relationships between scales from the Karolinska Scales of Personality (KSP) and the factor structure of the KSP in a sample of male juvenile delinquents. The KSP was administered to a group of male juvenile delinquents (n=55, mean age 17 years; standard deviation=1.2) from four Swedish national correctional institutions for serious offenders. As expected, the KSP showed appropriate correlations between the scales. Factor analysis (maximum likelihood) arrived at a four-factor solution in this sample, which is in line with previous research performed in a non-clinical sample of Swedish males. More research is needed in a somewhat larger sample of juvenile delinquents in order to confirm the present results regarding the factor solution. PMID- 16316898 TI - Evaluation of diagnostic procedures in Swedish patients with schizophrenia and related psychoses. AB - We aimed to estimate the value of structured interviews, medical records and Swedish register diagnoses for assessing lifetime diagnosis of patients with schizophrenia. Psychiatric records and diagnostic interviews of 143 Swedish patients diagnosed by their treating physician with schizophrenia and related disorders were scrutinized. Based on record analysis only, or a combined record and interview analysis, DSM-IV diagnoses were obtained by the OPCRIT algorithm. Independent of the OPCRIT algorithm, a standard research DSM-IV diagnosis, based on both record and interview analysis, was given by the research psychiatrist. Concordance rates for the different psychosis diagnoses were calculated. DSM-IV diagnoses based on records only, showed a good to excellent agreement with diagnoses based on records and interviews. Swedish register diagnoses displayed generally poor agreement with the research diagnoses. Nevertheless, 94% of subjects sometimes registered with a diagnosis of schizophrenic psychoses (i.e. schizophrenia, schizoaffective psychosis or schizophreniform disorder) displayed a standard research DSM-IV diagnosis of these disorders. For patients in long term treatment for schizophrenia in Sweden, psychiatric record reviews should be optimal, cost effective and sufficient for assessment of lifetime research diagnoses of schizophrenia. For these patients a research interview adds little new information. The results further indicate that a Swedish register diagnosis of schizophrenic psychoses has a high positive predictive power to a standard research DSM-IV diagnosis of the disorders. It is concluded that for future Swedish large-scale genetic studies focusing on a broad definition of schizophrenia, it would be sufficient to rely on the Swedish register diagnoses of schizophrenic psychosis. PMID- 16316899 TI - The Need of Support and Service Questionnaire (NSSQ): a brief scale to assess needs in clients with long-term mental disabilities. AB - We present the Need of Support and Service Questionnaire (NSSQ), a new instrument developed to provide professionals in psychiatric care and community social services with a tool to assess needs of support and service in mentally disabled clients. The final version of the instrument was designed after comments from professionals in psychiatric care, community social services, and the clients and their relatives. A reliability study (test-retest, n=77), inter-rater (n=69) and a validity study (n=529) were performed. In the validity study, comparisons were done with the Camberwell Assessment of Need (CAN) scale, the Global Assessment of Functioning (GAF) scale and the Social and Occupational Functioning Assessment Scale (SOFAS). The test-retest and inter-rater reliability were 0.86 and 0.76, respectively. The percentage of agreement on the individual items ranged from 83.1% to 100.0% (test-retest) and 76.8% to 100.0% (inter-rater reliability). The corresponding kappa coefficients ranged from 0.63 to 1.00 (test-retest) and 0.33 to 1.00 (inter-rater reliability). The comparison between NSSQ and CAN items demonstrated high sensitivity and specificity as well as satisfactory likelihood ratios (LRs). The correlations between GAF and SOFAS scores and the number of needs of support, number of needs of service and the total number of needs of support and service were -0.49, -0.32 and -0.47, and -0.48, -0.26 and -0.45, respectively. All correlations were statistically significant (P<0.01). The results demonstrated that the psychometric characteristics of the NSSQ were satisfactory. Moreover, professionals without any formal training easily used it. PMID- 16316900 TI - Sense of coherence among Finnish young men--a cross-sectional study at military call-up. AB - The aim of this study was to study associations between sense of coherence (based on Antonovsky's theory), and mental health problems, the use of mental health services, substance use, predisposition to suicide and possible psychiatric diagnosis among Finnish young men. The study included 2314 18-year-old young men who attended military call-up in autumn 1999 and completed the 13-item Orientation to Life Questionnaire (SOC-13) questionnaire (89.1% of the study sample attending military call-up). Furthermore, all young men had a medical examination at military call-up, including evaluation of possible psychiatric disorder. Suicidality, perceived mental health problems, psychiatric diagnosis at military call-up medical examination, use of mental health services, excessive alcohol consumption and use of illicit drugs were identified as independent factors associated with sense of coherence. Sense of coherence is a valuable construction in psychiatric evaluation of adolescents. SOC-13 can be used both in clinical practice and in epidemiological research to evaluate the adolescent's health resources. Future research on its applications in clinical practice is warranted. PMID- 16316901 TI - Have psychiatric problems of children in children's homes increased? AB - This time-trend study was conducted during and after the severe economic recession and the period of high unemployment in Finland in the 1990s. The changes in the behavioral and emotional problems of children and adolescents placed in children's homes were studied in the same area using the Child Behavior Checklist questionnaire at three different time points (1993, 1996 and 1999), and the Children's Global Assessment Scale in 1996 and 1999. Internalizing symptoms of the children increased significantly. The level of psychosocial problems among boys was already high at the first rating in 1993, and it remained high during the whole study period. Behavioral and emotional problems of the girls placed in children's homes increased significantly, being at the same level as the boys' by the end of the study period. PMID- 16316902 TI - Major depressive episode related to long unemployment and frequent alcohol intoxication. AB - We studied the association between two major problems--unemployment and major depressive episode--and the impact of different timing of periods of unemployment and risk factors, especially alcohol intoxication, for major depressive episode among the unemployed. Major depressive episode during the last 12 months, plus current and past employment status and frequency of alcohol intoxication, were assessed within the nationally representative, cross-sectional 1996 Finnish Health Care Survey, in which non-institutionalized individuals aged 15-75 years were interviewed by using the Short Form of the University of Michigan version of the Composite International Diagnostic Interview (the UM-CIDI Short Form). Of the 5993 subjects interviewed, 3818 (64%) were occupationally active and included in the logistic regression analysis, showing that even after adjusting for other potentially confounding variables, current unemployment was associated with major depressive episode (odds ratio, OR=1.78, 95% confidence interval, CI, 1.38-2.29). Further analysis revealed that the increased risk of major depressive episode was only related to long-term unemployment. Frequent alcohol intoxication (at least once a week) increased the risk of major depressive episode remarkably. Compared with the group "Constantly employed, no frequent alcohol intoxication", long-term unemployment with no frequent alcohol intoxication had moderately increased risk of major depressive episode (OR=1.72 (95% CI 1.29-2.30) and those with frequent alcohol intoxication had highly increased risk [OR=11.27 (95% CI 5.51-23.09) vs. OR=1.72 (95% CI 1.29-2.30]. Long-term unemployment is associated with increased risk of major depressive episode. Frequent alcohol intoxication among long-term unemployed individuals greatly increases the risk of depression. PMID- 16316903 TI - Effects of long-term benzodiazepine medication. A prospective cohort study: methodological and clinical aspects. AB - Benzodiazepine (BDZ) drug impact on mental functions was explored in a cohort study of 30 psychiatric outpatients on long-term BDZ medication. A new questionnaire, the Drug Impact on Mental Processes (DIMP) was used and evaluated. The patients were rated three times: on inclusion in the study, after about 18 months and, finally, only a few days later. Test-retest reliability was evaluated for the two last ratings and was found acceptable for 19/23 items. The DIMP scores indicated negative effects on crisis reaction, intensified defence mechanisms and reduced cognitive, emotional and conative functions. Long-term BDZ treatment intensified passive coping. The drug impact on mental functions ranged between a mild and a moderate degree. The clinical outcomes of the patients' psychiatric disorders were evaluated at a 1-year follow-up after continued or discontinued BDZ treatment. In the subgroup that had discontinued the BDZ treatment at follow-up, significantly more patients reported reduced severity of their psychiatric disorders and significantly more patients had paid jobs. The overall clinical improvement after discontinued BDZ treatment may be explained by recovery from an addiction syndrome. It may also be related to a shift from passive to active coping. PMID- 16316904 TI - Assessment of early mother-child relation in infants with oesophageal atresia. AB - Fifteen one-year old infants with oesophageal atresia and their parents were used to demonstrate how the Parent-Child Early Relational Assessment (PCERA) can be used to explore the mother-child interaction in a clinical sample of children. The infants and their mothers were videotaped for 5-min episodes during feeding and free play, and the quality of the mother-infant interactions was assessed by PCERA. The results showed areas of strength regarding most of the maternal, infant and dyadic components. There was concern about maternal positive physical contact, maternal quality of verbalizations to or about the child, maternal social initiative, infant's communicative competence and infant's visual contact with mother. This concern of social proximity between infants and their mothers was most prominent in the feeding situation. The present study indicates that oesophageal atresia in infants may constitute stressors impinging on the parent infant relationship. PCERA was found to be an adequate assessment tool with acceptable reliability to evaluate the mother-infant interaction. PMID- 16316905 TI - The prevalence of abuse in Swedish female psychiatric users, the perpetrators and places where abuse occurred. AB - The aims of the study were to investigate self-reported physical, sexual, emotional and economical abuse in Swedish female users of psychiatric services, who the perpetrators were and in which places abuse occurred. An anonymous self administrated questionnaire was answered in the waiting room of the services. The drop-out rate was 21% and n=1382 women completed the questionnaire. Fifty-three per cent of the women had been abused during childhood, 63% during adulthood and 31% during past year. Seventy-four per cent of those exposed during childhood were also exposed later in life. Women subjected to abuse reported longer contact with psychiatric care. Regardless of life period, the majority reported multiple and frequent abuse. Emotional abuse was most frequent reported in both childhood and adulthood followed by physical and sexual abuse. The reported perpetrators were mainly male persons to whom the woman had an intimate relationship. Mostly the abuse occurred in the women's own home. However, other women, strangers, acquaintances and relatives were also stated as perpetrator and abusive acts also took place in other homes, outdoors or down town. The high prevalence of abuse and its multiplicity point to the necessity for the care and support system to prioritize abuse against women with psychiatric illness. PMID- 16316906 TI - Locked entrance doors at psychiatric wards--advantages and disadvantages according to voluntarily admitted patients. AB - Entrance doors at wards where psychiatric care is provided are sometimes locked, which is not the case at wards where somatic care is provided. How locked entrance doors at psychiatric wards are experienced by patients has been investigated to a very limited extent. The aim was to describe voluntarily admitted patients' perceptions of advantages and disadvantages about being cared for on a psychiatric ward with a locked entrance door. Audio-taped, semi structured interviews were conducted with 20 patients voluntarily admitted at psychiatric wards. Content analysis revealed six categories of advantages and 11 categories of disadvantages. Most advantages were categorized as "protects patients and staff against 'the outside' ", "provides patients with a secure and efficient care" and "provides staff with a sense of control over the patients". Most disadvantages were categorized as "makes patients feel confined", "makes patients feel dependent on the staff" and "makes patients feel worse emotionally". Patients perceive a variety of advantages and disadvantages, for themselves, their visitors and staff, connected to locked entrance doors at psychiatric wards. A locked door may make the ward appear as both a prison and a sanctuary. It is important that staff try to minimize patients' concerns connected to the locked door. PMID- 16316907 TI - Ethics and interpreting in psychotherapy with refugee children and families. AB - Basic ethical principles like autonomy, non-maleficence, beneficence and justice should be taken into consideration step by step when treating refugee children and their families. These principles may be considered from the point of view of each of the actors involved--patient, therapist and interpreter. This paper is focused on the role of the interpreter and on different aspects to be considered by the therapist when working with interpreters in psychotherapeutic treatment of refugee children and families. Elements of case histories are used to illustrate situations faced in working with an interpreter. An ethical analysis of a case where a teenage refugee received therapeutic treatment using an interpreter is made. PMID- 16316908 TI - Should catatonia be part of the differential diagnosis of coma? AB - The descriptions of catatonia are complex and unclear. More than 40 different signs and symptoms have been included, but coma is not among them. Recently, a case report of coma in relation to catatonia was presented. We report two further cases where coma was a part of this syndrome of disturbed motility. They were successfully treated with ECT. Consequently, we suggest that catatonia may be a differential diagnosis of coma. PMID- 16316909 TI - Recorded psychiatric comorbidity with bipolar disorder--a Finnish hospital discharge register study. AB - Bipolar disorder is frequently connected to other psychiatric disorders. On the basis of The National Hospital Discharge Register in Finland, we studied the recorded prevalence of psychiatric comorbidity among bipolar inpatients by clinicians, and the factors that were associated with it. Of the 2687 hospital stays in 1998, 82% had no other recorded psychiatric diagnosis except an episode of bipolar disorder. Psychiatric comorbidity was recorded in 18% of hospital stays, of which 20% had two comorbid psychiatric diagnoses. Substance-related disorders (11%) were the most commonly recorded comorbid disorders. Personality disorders were recorded in 6%, and anxiety disorder in 1% of the hospital stays. These figures should be considered far below the expected ones. Recorded comorbidity was associated with the type of episode. Comorbidity in bipolar disorder in psychiatric hospitals in Finland seems to go greatly undetected and may have a deteriorating impact on the course of the illness. PMID- 16316910 TI - Mental disorders and DSM-IV paedophilia in 185 subjects convicted of sexual child abuse. AB - The objective of this study was to explore how sexual child abusers who meet the DSM-IV criteria for paedophilia differ from those who do not. We studied DSM-IV disorders and paedophilic interests in all cases of sexual child abuse referred to forensic psychiatric investigation in Sweden between 1993 and 1997 (n=185). Frequency and severity of other mental disorders did not differ between subjects with and without DSM-IV paedophilia. However, men with paedophilia had more previous paedophilia convictions, same-sex and younger victims and less often intoxicated status when committing the act. The concept of paedophilia as a mental disorder is not supported by the DSM-IV diagnosis, which primarily describes the acting out of attraction. PMID- 16316911 TI - Venlafaxine and asthma. AB - A 54-year-old man of Asian origin with major depression developed an asthma-like reaction during venlafaxine treatment. Two weeks after therapy was initiated, he experienced gradually worsening dry cough at night and periodically dyspnea during the daytime. After 5 weeks, clinical examination revealed marked signs of pulmonary obstruction and the forced expiratory volume (FEV1) was assessed to only 32% of the expected value. The venlafaxine medication was gradually decreased and eventually discontinued 9 weeks after its initiation, resulting in a successive improvement of the patient's respiratory complaints. PMID- 16316912 TI - The association between birth weight, season of birth and latitude. AB - BACKGROUND: Birth weight is known to fluctuate with month of birth and with latitude; however the mechanisms underlying these gradients remain unclear. AIM: The study examined within-year fluctuations in birth weight across four regions in Australia. SUBJECTS AND METHODS: We obtained the birth weight of singleton births of at least 37 weeks gestation from four regions: (a) southeast Queensland, (b) Sydney and surrounding districts, (c) Victoria, and (d) Tasmania. Seasonal fluctuations were examined after long-term trends had been removed. The amplitude, timing and variability of seasonal fluctuations were examined and compared across regions. RESULTS: Each of the four regions had prominent annual periodicity with the peak birth weight occurring in spring. The amplitude of this signal ranged from 1.4 g (Victoria) to 7.7 g (southeast Queensland). There was no significant correlation between the amplitude of these fluctuations and the latitude of four regions. In addition, a second smaller autumn peak was identified. CONCLUSION: Environmental factors that have regular within-year variation influence birth weight in Australia. The evidence suggests that at least two distinguishable seasonal processes contribute to the findings. Identifying the factors underlying these seasonal fluctuations could have implications from a public health perspective. PMID- 16316913 TI - The spectrum of haemoglobin variants in two scheduled tribes of Sundargarh district in north-western Orissa, India. AB - BACKGROUND: Tribal communities in India constitute a major part of the population and are vulnerable to many erythrocytic hereditary and haematological disorders such as haemoglobinopathies. Genetic studies so far undertaken on tribal groups are scanty, patchy and incomplete. No field-based systematic studies of hereditary haemolytic disorders in Orissa are available. Further, the extent of haemoglobin variants among the tribals in the state is not known. The present study was carried out in the Bhuyan and Kharia tribes of Sundargarh district in Orissa. AIM: This study aims to find the prevalence/spectrum of haemoglobin variants in two major tribal groups, namely Bhuyan and Kharia and their subgroups, inhabiting the Sundargarh district in north-western Orissa. SUBJECTS AND METHODS: Following the probability proportionate to size cluster sampling procedure for villages, a randomized sampling procedure was adopted irrespective of the age, sex and individual susceptibility pattern, selecting exclusive villages of each sub-group of tribes in five blocks. A total of 1603 blood samples of 836 Bhuyan and 767 Kharia tribals were screened for haemoglobin variants in the Sundargarh district of Orissa. Laboratory analyses of blood samples were carried out following standard procedures. RESULTS: The study showed a high prevalence of haemoglobin variants in the Bhuyan (9.8%) and Kharia (13.3%) tribes, sickle-cell disorders contributing 2.4% and 5.6%, respectively. The sickle-cell gene was found to be completely absent in the Dudh Kharia tribe, whereas the frequency in the Dhelki Kharia was quite high (12.5%). For the first time, 1.4% prevalence of haemoglobin E disorders (10 traits and one disease case) was recorded in a tribal population, i.e. Delki Kharia in Orissa. No other haemoglobin variant except beta-thalassaemia trait was detected in the Dudh Kharia tribe (8.1%), showing their genetic isolation (p < 0.001) from the Delki Kharia (4.1%), the average being 6.3% in the Kharia tribe. Out of three subgroups of Bhuyan studied, the sickle-cell trait was detected only in Paraja (0.9%) and Paik (7.4%), and not in Paudi (Hill) Bhuyans. However, the beta-thalassaemia trait was detected in an average 6.5% in the Bhuyan tribe: in Paudi (2.1%), Paik (7.8%) and in Paraja (12.7%). For the first time in the tribes of Orissa a family was found with haemoglobin D trait (in Paik Bhuyan) and another with hereditary persistence of fetal haemoglobin (in Paraja Bhuyan). Clinical and haematological features of these disorders were similar to those reported in previous studies carried out in India. CONCLUSION: Isolates of the Bhuyan and Kharia tribes show intra-group variations in prevalence of haemoglobin variants due to founder effect, genetic drift, and the practice of inbreeding in varied geographical and ecological niches in the Sundargarh district of Orissa. PMID- 16316914 TI - Indian ethnic populations characterized by dopamine (D4) receptor VNTR polymorphism. AB - BACKGROUND: The human dopamine receptor D4 gene (DRD4) contains a 48-bp tandem repeat in exon 3 and shows alleles varying between repeats 2 and 11. The gene shows a high level of expression in the prefrontal cortex of the brain and association of particular alleles of this locus with various neuropsychiatric and personality disorders have been reported. OBJECTIVE: The present study reports allele frequency distribution at the DRD4 variable number tandem repeat (VNTR) locus among five ethnic populations of India. This background information is fundamental to the field of pharmacogenetics for disease susceptibility and association studies. SUBJECTS AND METHODS: Three hundred and thirty two healthy unrelated adult individuals belonging to five ethnic groups: Konkanastha Brahmins, Marathas, Ezhavas, Nairs and Muslims, have been typed. Genomic DNA, extracted from peripheral blood, was PCR amplified using a two-enzyme system. The use of ALF Express DNA sequencer was found to be helpful for large-scale population genotyping. Statistical analysis was performed using the POPGENE and DISPAN programs. RESULTS: A total of eight alleles ranging from repeat 2 to repeat 9 were observed. Allele 4 was the predominant allele among all the five populations, consistent with the data on other world populations. A rare allele 9 was detected exclusively among Marathas. The observed heterozygosity was low, ranging from 0.38 to 0.54 while other parameters like Polymorphism Information Content (PIC) and Power of Discrimination (PD) showed moderate values. The populations were in genetic equilibrium when tested under Hardy-Weinberg expectations. CONCLUSION: The allele frequency estimates for DRD4 provided here will contribute towards developing an informative database for this functionally relevant locus. This will prove useful when studying the association between genetic factors and pathogenesis of disease in Indian populations and will address the concern of biased results of association due to population admixtures. PMID- 16316915 TI - Quantitative genetic study of head size related phenotypes in ethnically homogeneous Chuvasha pedigrees. AB - BACKGROUND: It is well established that genetic factors contribute significantly to the determination of head size and shape traits variability. However, the controversies in views and findings with respect to the more specific aspects of this issue have not yet been resolved. AIM: The primary objective of the study was to examine the patterns of the intergenerational familial transmission of 12 head size related traits in a large ethnically homogeneous sample of Chuvasha pedigrees. SUBJECTS AND METHODS: The research was carried out on 1406 individuals belonging to 357 nuclear and more complex families. Univariate and bivariate family-based analyses were performed to establish the pattern of head traits inheritance. RESULTS: Maximum heritability estimates ranged from 0.52 to 0.72 for traits adjusted for significant covariates. No significant sex differences were observed with respect to the genetic determination of the studied traits. Bivariate analysis of horizontal and vertical head size components suggested the existence of common genetic and environmental factors that explained 33.0% and 23.2% of the total variance of the adjusted traits, respectively. CONCLUSIONS: A significant genetic component is involved in inter-individual variation and covariation of various studied craniofacial traits. PMID- 16316916 TI - Anthropometry, lifestyles and fat patterning in Balinese women. AB - BACKGROUND: In many parts of the developing world, modernization has resulted in an increase in the prevalence of overweight and obesity and a subsequent rise in nutrition-related, non-communicable diseases such as cardiovascular disease and Type 2 diabetes. AIM: The study examined the impact of socio-economic and lifestyle changes associated with modernization on the body size and shape of Balinese women. SUBJECTS AND METHODS: Anthropometric data including weight, height, body mass index (BMI) and seven skinfolds were collected from 564 Balinese women, aged between 15 and 45 years. The relationship between the degree of 'modernization' (using such parameters as household wealth and education level) and body size and shape was analysed using multiple regression analysis, as was the effect of age, family size and breast-feeding. RESULTS: Using the WHO (WHO 1995) categories of BMI, 14% of Balinese women displayed Chronic Energy Deficiency (BMI < 18.5 kg m(-2)), 7% were overweight (BMI = 25.0-29.9 kg m(-2)) and less than 1% were obese (BMI > or = 30.0 kg m(-2)). Household wealth (p < 0.0001) and education level (p < 0.05) were significantly associated with fatness, while breast-feeding (p < 0.05) was significantly associated with leanness. Fat distribution was largely independent of biological and socio economic variables. CONCLUSIONS: The trend toward increasing fatness associated with greater household wealth and higher education has important health implications for Balinese women as they continue their rapid transition from subsistence agriculture towards a more 'western' lifestyle. PMID- 16316917 TI - Genetic variations of the apolipoprotein B gene in Turkish patients with coronary artery disease. AB - BACKGROUND: The results of studies that clarify the association of genetic markers at the apolipoprotein B (apo B) gene (EcoRI and XbaI polymorphisms) with coronary artery disease (CAD) are not consistent and suggest that the effect is context dependent (dependent on ethnicity and sex). The present study represents the first investigation of the apo B gene polymorphisms in Turkish patients with CAD and their influence on lipid levels. AIM: The study investigated the association of apo B gene EcoRI and XbaI polymorphisms with CAD and with variation in lipid levels (total cholesterol (T-Chol), high-density lipoprotein cholesterol (HDL-Chol), low-density lipoprotein cholesterol (LDL-Chol), and triacylglycerol (TAG)). SUBJECTS AND METHODS: The study group was composed of 150 individuals with angiographically documented CAD and 100 angiographically proven to be healthy controls. PCR-RFLP was used to determine the DNA polymorphisms of the apo B gene. RESULTS: The frequencies of apo B genotypes detected with EcoRI (AA, AG, GG) and XbaI (CC, CT, TT) did not differ significantly between case and control subjects. A significant association between EcoRI genotypes and T-Chol (p < or = 0.05), and LDL-Chol (p < or = 0.001) was observed only in CAD patients. Patients with the AA genotype had higher levels of serum T-Chol and LDL-Chol compared with AG. With logistic regression analysis the XbaI TT genotype was found to be associated with CAD prevention. However, no significant differences in lipid variables were determined for the XbaI polymorphisms in the patients with CAD. CONCLUSIONS: Apo B EcoRI genotypes were not found as risk factors for CAD, whereas XbaI TT genotype was detected to prevent against CAD in our study group. PMID- 16316918 TI - Menarcheal age and socio-economic factors in Poland. AB - BACKGROUND: Menarcheal age is the most important measure of sexual maturation in girls and a sensitive indicator of environmental conditions during childhood. OBJECTIVE: The study analysed the association between age at menarche and socio economic characteristics (urbanization, population size, education of parents and number of children in the family). SUBJECTS AND METHODS: Questionnaire data were collected from 3271 female schoolchildren born between 1981 and 1984, living in three provinces of southern Poland. Menarcheal age was estimated by the recall method and based on the date of menarche given by the study participants. ANCOVA and multiple regression analyses were applied to test statistical significance of differences between groups. RESULTS: Girls from families with high socio-economic status experience menarche at an earlier age than girls from families with lower socio-economic status. However, depending on the geographical region and the population size, other factors influence menarcheal age. In the Krakow province, factors that significantly differentiate age at menarche are urbanization, father's education and number of children in the family; in the Opole province, these factors include urbanization and number of children in the family, while in the Nowy Sacz province, number of children in the family is significant. CONCLUSIONS: Socio-economic differences are greater in a large urban city (Krakow), and affect variation in age at menarche. However, within smaller populations (Opole, Nowy Sacz) living in towns and villages, the difference in age at menarche is less visible. In addition, variation between areas reveals a lower age at menarche in urban as compared with non-urban areas. PMID- 16316919 TI - Anthropometry of Malawian live births between 35 and 41 weeks of gestation. AB - The aim of this analysis was to construct cross-sectional gestational age specific percentile curves for birthweight, length, head and mid-arm circumference for Malawian babies, and to compare these percentiles with reference values for babies born to women with normal pregnancies, from a developed country. A cross-sectional study which enrolled pregnant women attending two study hospitals between March 1993 and July 1994 was undertaken. Data on maternal socio-economic status, newborn anthropometry, previous obstetric history and current pregnancy were collected. Smoothed percentile values were derived using the LMS method. Malawian reference percentiles were constructed for fetal growth from 35 weeks' gestation for singleton births. Mean birthweight, length and head circumference were lower at all gestational ages for Malawian compared with Swedish newborns. Fetal growth per completed gestational week was higher by 60 g in weight, 0.5 cm in length and 0.2 cm in head circumference in Swedish compared with Malawian babies. Growth restriction was present from 35 to 41 weeks' gestation. The pattern for the 10th percentile suggested that this was occurring from well before 35 weeks' gestation in a proportion of babies. PMID- 16316920 TI - Secular growth changes in body height and weight in children and adolescents in Shandong, China between 1939 and 2000. AB - BACKGROUND: There is strong evidence of a positive secular trend in physical growth in most of the world during the last century. However, no studies on this trend have been reported in Shandong Province, China. AIM: The study assessed the secular trend in body height and weight in Shandong Province during the past few decades and the association of socio-economic status with the intensity of the trend. SUBJECTS AND METHODS: The results of height and weight in 7-18-year-old boys and girls obtained in nine subsequent surveys (1939, 1956, 1962, 1972, 1979, 1985, 1991, 1995, and 2000) were included in this analysis. The differences of mean values between surveys and the increments per decade were compared. Socio economic indicators were collected and analysed. RESULTS: In 18-year-old groups, mean height increased from 164.9 to 172.6 cm for boys and from 155.9 to 160.6 cm for girls, corresponding to 1.75 and 1.07 cm per decade, respectively, and the increments of weight were 7.5 kg for boys and 0.3 kg for girls, corresponding to 1.70 and 0.07 kg per decade, respectively, between 1956 and 2000. The biggest changes of physical growth were in 13-year-old boys and 11-year-old girls. CONCLUSION: Positive secular trends occurred in 7-18-year-old children and adolescents in Shandong Province, China during 1939-2000, which may reflect the secular changes in socio-economic status. PMID- 16316921 TI - COL1A2 (type I collagen) polymorphisms in the Colorado Indians of Ecuador. AB - BACKGROUND: EcoRI, MspI and RsaI restriction fragment length polymorphisms (RFLPs) of the COL1A2 (type I collagen) gene are proving to be extremely informative markers for describing human populations; therefore they hold considerable potential for anthropogenetic research. AIM: The objective of this study was to characterize at the DNA level the Colorado Indians from Ecuador, for whom only blood group frequency information is available, and to investigate their relationships with the Cayapa-another Ecuadoran Native American group belonging to the same linguistic affiliation-and other world populations. SUBJECTS AND METHODS: Colorado Indians (n = 80) were analysed for the three anthropologically informative RFLPs of the COL1A2 gene. To better define the genetic relationship between this group and other populations, principal component analysis (PCA) was performed and genetic distances were estimated. Population genetic structure was tested through analysis of molecular variance (AMOVA) by comparing haplotype frequencies. RESULTS: COL1A2 allele and haplotype frequencies showed a certain degree of heterogeneity between the two Chibchan populations of Ecuador. The AMOVA test detected a significant level of differentiation (Fst = 0.034, p = 0.0049) between Colorado and Cayapa Indians. PC and genetic distance analyses showed a clear-cut separation between African and non-African populations; within the latter, the two Native American groups were differentiated from each other. CONCLUSIONS: The present findings suggest the presence of a low level of genetic relatedness between the Colorado and the Cayapa, despite their supposed common ethnogenesis. This confirms what has been inferred from other genetic data about the high degree of heterogeneity among Native Americans, even within the same linguistic branch, thus supporting the existence of genetic sub-structure within the central and southern American populations. PMID- 16316922 TI - The effect of variability in maturational tempo and midparent height on variability in linear body measurements. AB - BACKGROUND: Information on the effect of variability in maturational tempo on variability in height attained at the same age is not readily available. AIM: The study obtained this information from the data of the First Zurich Longitudinal Study. SUBJECTS AND METHODS: Yearly measurements of standing height, sitting height and leg length, yearly assessments of bone age (RUS (Radius, Ulna and Short bones), TW3 method) and midparent height for 232 children from the First Zurich Longitudinal Growth Study are included in a correlational analysis. RESULTS: The course of the squared correlations of standing height, sitting height and leg length with attained RUS bone age, midparent height and both as a function of age are presented. CONCLUSIONS: During puberty, up to 50% of the height variation in boys and 40% in girls is explained by maturational tempo. PMID- 16316926 TI - CYP2A13 genetic polymorphism in French Caucasian, Gabonese and Tunisian populations. AB - Since human CYP2A13 is expressed in the respiratory tract and is involved in the activation of tobacco-specific nitrosamines, some of the previously reported sequence variations may contribute to inter-individual and inter-ethnic differences in the susceptibility of tobacco-related tumorigenesis. The aim was to compare the frequencies of the 578C > T (Arg101Stop), 3375C > T (Arg257Cys) and 7520C > G (3'-untranslated region) mutations in several populations. The frequencies of the 578C > T polymorphism were 3.8, 0 and 1.0% in French Caucasians, Gabonese and Tunisians, respectively. In the same populations, the frequencies of the 3375C > T mutation were 0, 15.3 and 4.2%, respectively, whereas the frequencies of the 7520C > G mutation were 1.0, 20.8 and 7.3%, respectively. Marked inter-ethnic variations in CYP2A13 were identified and confirmed. These findings provide data for further studies that associate CYP2A13 haplotypes with an incidence of smoking-related tumours in respect of ethnicity. PMID- 16316927 TI - Effect of polymeric black tea polyphenols on benzo(a)pyrene [B(a)P]-induced cytochrome P4501A1 and 1A2 in mice. AB - The chemopreventive activity of green tea polyphenols (GTPs) is, in part, due to modulation of cytochrome P450s (CYPs). To investigate the enzyme modulatory properties of major black tea polyphenols, the effect of decaffeinated black tea extract (DBTE) or polymeric black tea polyphenol (PBP) mix was studied on CYP1A1 and CYP1A2 in mouse tissues. Animals receiving 2.5% DBTE or 1% PBP mix or drinking water (15 days) were challenged with single oral benzo(a)pyrene (B(a)P) (1 mg/mouse) treatment on the 14th day. Liver and lung microsomes isolated after 24 h were analysed for CYP1A1 and CYP1A2, using biochemical substrate(s) and Western blot analysis. Treatment with 2.5% DBTE or 1% PBP mix did not significantly alter the basal activity and level of CYP1A1 and CYP1A2, whereas pretreatment with 2.5% DBTE or 1% PBP mix resulted in a significant decrease in both the activity and the level of B(a)P-induced CYP1A1 and CYP1A2 in liver and lungs. The PBP mix possesses enzyme modulatory properties exhibited by monomeric GTPs. PMID- 16316928 TI - Comparative in vitro metabolism of methoxychlor in male and female rats: metabolism of demethylated methoxychlor metabolites by precision-cut rat liver slices. AB - The in vitro metabolism of demethylated methoxychlor (MXC) metabolites, mono-OH MXC (including (R)- and (S)-isomers) and bis-OH-MXC (mono- and bis-demethylated MXC, respectively), was conducted using precision-cut liver slices to understand the sex-dependent metabolism of MXC in rats. In the study with bis-OH-MXC, the substrate underwent extensive conjugation producing its glucuronide and glucuronide/sulphate diconjugate, and no significant sex differences were found. On the contrary, the metabolism of mono-OH-MXC appeared to exhibit the sex differences in the metabolic profiles. The bis-OH-MXC glucuronide and glucuronide/sulphate diconjugate were major metabolites in male rat, whereas the mono- and bis-OH-MXC glucuronides predominated in the female. The per cent distribution of the demethylated products (sum of bis-OH-MXC derivatives) was approximately 90% for the male (for both isomers) and 81 (R-) to 56% (S-) for the female. The metabolic profiles in (S)-mono-OH-MXC, which is the predominant enantiomer preferentially produced in MXC metabolism in rats, showed a similar pattern to that of MXC compared with the (R)-isomer. The results indicate that the sex differences in oxidative demethylation of the intermediate, (S)-mono-OH MXC, could be one of the probable reasons for the sex-dependent metabolism of MXC in rats, and the stereo-structural preference of the contributing demethylase enzymes appear to be involved. PMID- 16316929 TI - Metabolic activation of a pentafluorophenylethylamine derivative: formation of glutathione conjugates in vitro in the rat. AB - The aim was to investigate the metabolic activation potential of a pentafluorophenylethylamine derivative (compound I) in vitro in the rat and to identify the cytochrome P450 (CYP) enzymes that catalyse these metabolic activation processes. Reduced glutathione (GSH) was fortified in rat hepatocytes and liver microsomes to trap possible reactive intermediates. Four glutathione conjugates (M1-4) were identified by LC-MS(n) following incubation of compound I in GSH-enriched rat hepatocytes and liver microsomes. Three of these conjugates (M2-4) have not been reported previously for pentafluorophenyl derivatives. Elemental composition analysis of these conjugates was obtained using high resolution quadrupole time-of-flight mass spectrometry. The formation of GSH conjugate M1 was rationalized as a direct nucleophilic replacement of fluoride by glutathione, whereas the formation of the GSH conjugates M2-4 was proposed to occur by NADPH-dependent metabolic activation of the pentafluorophenyl ring via arene oxide, quinone and/or quinoneimine reactive intermediates. Formation of these conjugates was enhanced three- to five-fold in liver microsomes obtained from phenobarbital- and dexamethasone-treated rats. In incubations with pooled rat liver microsomes and recombinant rat CYP3A1 and CYP3A2, troleandomycin (TAO) reduced the formation of GSH conjugates M2-4 by 80-90%, but it had no effect on the formation of M1. Incubation of compound I with rat supersomes indicated that only CYP3A1 and CYP3A2 were capable of mediating these metabolic activation processes. PMID- 16316930 TI - Kinetic studies on the intramolecular acyl migration of beta-1-O-acyl glucuronides: application to the glucuronides of (R)- and (S)-ketoprofen, (R)- and (S)-hydroxy-ketoprofen metabolites, and tolmetin by 1H-NMR spectroscopy. AB - Conjugation of carboxylate drugs with D-glucuronic acid is of considerable interest because of the inherent reactivity of the resulting beta-1-O-acyl glucuronides. These conjugates can degrade by spontaneous hydrolysis and internal acyl migration. beta-1-O-acyl glucuronides and their acyl migration products can also react covalently with macromolecules with potential toxicological consequences. The spontaneous degradation of the diastereoisomeric beta-1-O-acyl glucuronide metabolites of the racemic drug ketoprofen, two of its ring hydroxylated metabolites and of tolmetin beta-1-O-acyl glucuronide was investigated by (1)H-NMR spectroscopy in buffer solutions, at pH 7.4 and 37 degrees C. A plot of the logarithm of the peak integrals against time revealed first-order kinetics. Degradation rates and half-lives were calculated for each glucuronide using first-order reaction equations. Tolmetin glucuronide had the fastest degradation rate, whilst all of the ketoprofen-related glucuronides had similar degradation rates. The degradation of the diastereoisomeric glucuronides was stereoselective, with the rate for the (S)-isomer always slower compared with the (R)-isomer by approximately a factor of 2. PMID- 16316931 TI - Impaired glutathione-conjugating capacity by cryopreserved human and rat hepatocytes. AB - The activity of glutathione transferase was measured in sonicates of fresh rat hepatocytes and of cryopreserved rat, human and dog hepatocytes in the presence of added glutathione and by using 1-chloro-2,4-dinitrobenzene (CDNB) as non selective substrate. The glutathione-conjugating capacity was also investigated in the presence of CDNB alone (without glutathione) with intact fresh rat hepatocytes and cryopreserved rat and human hepatocytes. Finally, the intracellular level of glutathione was measured in these hepatocytes. The specific activity of glutathione transferase in sonicates of fresh rat hepatocytes (in the presence of added GSH and CDNB) was about 415 nmol/min/10(6) cells. The corresponding activities in cryopreserved rat, human and dog hepatocytes were approximately 320, 440 and 540 nmol/min/10(6) cells, respectively. In contrast, glutathione conjugation by the intact cryopreserved human and rat hepatocytes in the presence of CDNB alone was less than 10% of the corresponding conjugation by fresh rat hepatocytes, indicating that glutathione was depleted in these cryopreserved hepatocytes. Glutathione depletion was confirmed after analytical measurement of the glutathione levels in fresh and cryopreserved hepatocytes. In fresh rat hepatocytes the level of glutathione was 44 nmol/10(6) cells, whereas it was 2.5 and 4.4 nmol/10(6) cells in cryopreserved rat and human hepatocytes, respectively. In summary, glutathione transferase was active in these cryopreserved hepatocytes but the cryopreservation procedure likely causes depletion in the intracellular level of glutathione, resulting in an overall reduced glutathione conjugating capacity. PMID- 16316933 TI - The application of youth substance use media campaigns to problem gambling: a critical evaluation. AB - Despite the negative impact that problem gambling poses on individuals and society there have been few gambling prevention campaigns specifically targeting youth. The authors review the literature on past and current drug, alcohol, and tobacco use prevention media campaigns, examining the similarities across 25 health communication programs with the aim of viewing their applicability for the prevention of youth problem gambling. Critical features of effective campaigns are identified and assessed in terms of their applicability for youth problem gambling. Recommendations for the design, implementation, and evaluation of a youth gambling media campaign are discussed. PMID- 16316932 TI - Effects of fibrates on human organic anion-transporting polypeptide 1B1-, multidrug resistance protein 2- and P-glycoprotein-mediated transport. AB - The effects of different fibric acid derivatives (bezafibrate, clofibrate, clofibric acid, fenofibrate, fenofibric acid and gemfibrozil) on human organic anion transporting-polypeptide 1B1 (OATP2, OATP-C, SLC21A6), multidrug resistance protein 2 (MRP2/ABCC2) and MDR1-type P-glycoprotein (P-gp/ABCB1) were examined in vitro. Cyclosporin A (a known inhibitor of OATP1B1 and P-gp), MK-571 (a known inhibitor of MRP2) and cimetidine (an organic cation) were also tested. Bezafibrate, fenofibrate, fenofibric acid and gemfibrozil showed concentration dependent inhibition of estradiol 17-beta-D-glucuronide uptake by OATP1B1-stably transfected HEK cells, whereas clofibrate and clofibric acid did not show any significant effects up to 100 microM. Inhibition kinetics of gemfibrozil, which exhibited the most significant inhibition on OATP1B1, was shown to be competitive with a Ki = 12.5 microM. None of the fibrates showed any significant inhibition of MRP2-mediated transport, which was evaluated by measuring the uptake of ethacrynic acid glutathione into MRP2-expressing Sf9 membrane vesicles. Only fenofibrate showed moderate P-gp inhibition as assessed by measuring cellular accumulation of vinblastine in a P-gp overexpressing cell-line. Cyclosporin A significantly inhibited OATP1B1 and P-gp, whereas only moderate inhibition was observed on MRP2. The rank order of inhibitory potency of MK-571 was determined as OATP1B1 (IC50: 0.3 microM) > MRP2 (4 microM) > P-gp (25 microM). Cimetidine did not show any effects on these transporters. In conclusion, neither MRP2- nor P-gp-mediated transport is inhibited significantly by the fibrates tested. Considering the plasma protein binding and IC50 values for OATP1B1, only gemfibrozil appeared to have a potential to cause drug-drug interactions by inhibiting OATP1B1 at clinically relevant concentrations. PMID- 16316934 TI - Parent ads in the National Youth Anti-Drug Media Campaign. AB - The National Youth Anti-Drug Media Campaign aims not only to reduce drug use by teens and preteens, but also to arm parents with knowledge about specific parenting practices known to reduce the risk of teen drug use. Among the documented successes of the campaign to date was a small, but direct effect on some parenting practices, including parent-child discussions about drug use. To reach a deeper understanding about the substance of the parental ads, we content analyzed the message strategies employed in the campaign's parent ads over the inaugural 5 years of the campaign. Each ad was coded for its major theme, minor subtheme, and featured drug. Among seven possible major themes, the parental anti drug ads largely featured four: enhance the risk of their child's drug use, encourage monitoring practices, promote parent-child discussions about drug use, or advocate positive involvement behaviors. Moreover, most parental messages addressed marijuana use or addressed drug use in general. Marijuana and inhalant ads largely were risk based, while general drug messages focused on monitoring, parent-child discussions or positive involvement practices. PMID- 16316935 TI - Factors affecting trust in on-line prescription drug information and impact of trust on behavior following exposure to DTC advertising. AB - Despite growing concerns about the quality and accuracy of Internet-based prescription drug information, there has been very little empirical research on consumers' perceptions of the trustworthiness of on-line drug information. In this article, we report on a study modeled after that of Menon, Deshpande, Perri, and Zinkhan (2002) in Health Marketing Quarterly that reexamines how key demographic, predispositional, and media factors are associated with consumer trust in on-line prescription drug information and the impact of trust in on-line drug information on ad-promoted behavior following exposure to direct-to-consumer (DTC) advertising. Four major findings are reported: (1) on-line drug information is not highly trusted; (2) trust in on-line drug information is not differentially affected by consumer demographic or predispositional characteristics; (3) trust in the traditional media of DTC advertising is predictive of trust in on-line drug information; and (4) trust in on-line drug information is associated directly with specific types of ad-promoted behavior following exposure to DTC advertising. Implications and recommendations are offered based on the results. PMID- 16316936 TI - Designing and implementing E-health Applications in the UK's National Health Service. AB - Telemedicine/e-health applications have the potential to play an important role in Britain's National Health Service (NHS), including the NHS in Scotland. The Scottish Telemedicine Action Forum (STAF) was established by the Scottish Executive Department of Health in 1999 to take a range of applications, targeted on national priorities, into routine service. In the process it has provided insights into how advanced information and communication technologies (ICTs) can be moved from the research stage into routine service. In this article four of the projects are described and analysed focusing on the key issues that have emerged as critical for carrying projects successfully through to implementation in service as follows: 1. A multisite videoconferencing network linking 15 minor injury units to the main accident and emergency (A&E) centre. 2. A single-site neonatal intensive care "cotside" laptop system to assist communication between parents and clinical staff. 3. A single-site outpatient chronic disease management system. 4. A multisite software audit tool to support the care of cleft lip and palate patients from birth onward. PMID- 16316937 TI - Gene cuisine or Frankenfood? The theory of reasoned action as an audience segmentation strategy for messages about genetically modified foods. AB - Genetically modified (GM) foods are currently a controversial topic about which the lay public in the United States knows little. Formative research has demonstrated that the lay public is uncertain and concerned about GM foods. This study (N = 858) extends focus group research by using the Theory of Reasoned Action (TRA) to examine attitudes and subjective norms related to GM foods as a theoretical strategy for audience segmentation. A hierarchical cluster analysis revealed four unique audiences based on their attitude and subjective norm toward GM foods (ambivalent-biotech, antibiotech, biotech-normer, and biotech individual). Results are discussed in terms of the theoretical and practical significance for audience segmentation. PMID- 16316938 TI - Targeting adolescents? The content and frequency of alcoholic and nonalcoholic beverage ads in magazine and video formats November 1999-April 2000. AB - This study compared alcoholic and nonalcoholic beverage advertising to which adolescents are exposed. A census of beverage advertising (N = 757) in popular magazines and television during November 1999-April 2000 was analyzed. Most alcohol ads appeared in Sports Illustrated (110), Rolling Stone (98), and Playboy (75) and outnumbered nonalcoholic beverage advertising by 3 to 1. Alcohol was almost never associated with dining. Alcohol ads emphasized sexual and social stereotypes and lacked diversity. One of every 6 magazine alcohol ads, and 1 of every 14 video-based ads, appeared to target teenagers. Many similarities existed between alcohol and nonalcohol ads. We conclude that alcohol is advertised heavily to youth through placement and appeals. The fact that themes in alcohol ads frequently parallel those in nonalcoholic beverage ads may further increase youths' receptivity. PMID- 16316941 TI - Lack of urinary bladder carcinogenicity of sodium L-ascorbate in human c-Ha-ras proto-oncogene transgenic rats. AB - Sodium L-ascorbate (Na-AsA) is widely known to be a tumor promoter of rat bladder carcinogenesis but tests negative in standard 2-year bioassays. In the present study, bladder-cancer-susceptible transgenic rats designated Hras128 were used to further examine the tumorigenicity of Na-AsA. A total of 40 7-week-old male transgenic (Tg) and 42 littermate nontransgenic (Non-tg) rats were divided into 4 groups and given powdered MF diet with or without 5% Na-AsA for 57 weeks. Tg rats showed significantly short survival compared with Non-tg, independent of Na-AsA treatment. Tg rats treated with Na-AsA showed a slightly higher incidence of carcinoma (29.6%) as compared to those without Na-AsA treatment (15.4%), but this was without statistical significance. Moreover, the total bladder tumor incidences, including papillomas, did not differ statistically (with Na-AsA, 37.0%; without Na-AsA, 30.8%). No bladder tumor was detected in Non-tg rats. Various kinds of other lesions in various organs were noted in Tg rats treated with or without Na-AsA treatment, but no intergroup differences were evident. In conclusion, Na-AsA did not show tumorigenicity in highly bladder-cancer susceptible transgenic Hras128 rats. These results suggest that Na-AsA is a pure promoter but not a complete carcinogen in rats. PMID- 16316942 TI - Global gene expression profiling of chemically induced rat mammary gland carcinomas and adenomas. AB - Chemical carcinogens induce both benign and malignant mammary gland tumors in female Sprague-Dawley rats. To identify gene expression profiles associated with malignancy, cDNA microarray analysis was used to compare gene expression profiles in rat mammary gland carcinomas, adenomas, and normal mammary gland. Tumors were induced with various chemical carcinogens including 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5 f]quinoxaline (MeIQx), 7-12-dimethylbenz[a]anthracene (DMBA), N-nitrosomethylurea (NMU), and 4-aminobiphenyl. The global gene expression profiles in carcinomas and adenomas were distinguishable by hierarchical clustering and multi-dimensional scaling analyses. Permutation analysis revealed 110 clones statistically differentially expressed between benign and malignant tumors (p < 0.0005). Carcinomas showed relatively high expression of several genes associated with mammary epithelial cell growth and proliferation (e.g., cyclin D1, PDGFalpha) and relatively low expression of differentiation marker genes (e.g., beta -casein, whey acidic protein, transferrin). Other categories of genes showing differential expression between carcinomas and adenomas were associated with protein homeostasis, cytoskeleton, extracellular matrix, and cell metabolism (fatty acid metabolism, oxidative phosphorylation, and glycolysis). Major gene families implicated in malignancy by over-expression in carcinomas included the annexins (annexin A1 and A4) and Stat family of transcription factors (Stat3 and Stat5a). The elevated expression of the prolactin receptor in carcinomas concomitant with several components of the mitogenic prolactin signaling pathway implicated prolactin/prolactin receptor/Stat5a/cyclin D1 in rat mammary gland malignancy. PMID- 16316944 TI - Electron microscopy in neuromuscular disorders. AB - Electron microscopy has a strategic position in the diagnosis of neuromuscular disorders. In muscular fibers, the main abnormalities include vacuoles, inclusion bodies, and myofibrillar disorganization with or without abnormal inclusion material. Vacuolar changes include lipidic and glycogenic storage vacuoles, rimmed vacuoles, and lysosomal and autophagic vacuoles. Accumulation of abnormal inclusion material is found in nemaline myopathy, actinopathies, and hyaline body myopathy. Myofibrillar disorganization involves cores, multiminicores, and myosin chain depletion. Myofibrillar myopathies associate a pathologic pattern of myofibrillar dissolution and ectopic protein expression. They can be divided into two groups: myofibrillar myopathies with multiple expression proteins and myofibrillar myopathies with desmin and alphaB-crystallin expression only. In these two conditions, electron microscopy shows accumulation of a granulofilamentous material immunoreactive for desmin. At least three genes are implicated: desmin, alphaB-crystallin, and myotilin. Lastly, electron microscopy serves to identify changes, pathogenic or not, which are not shown up by light microscopy. Moreover, electron microscopy gives insight on pathophysiological mechanisms and can guide molecular genetics analysis. PMID- 16316945 TI - Peritubular capillary changes in alport syndrome, diabetic glomerulopathy, Balkan endemic nephropathy and hemorrhagic fever with renal syndrome. AB - The morphology of peritubular capillary has been mostly studied in relation to chronic transplant rejection, where an association has been found between transplant glomerulopathy and reduplication of peritubular capillary basement membranes (PCBM). This electron microscopy study of peritubular capillaries was done on kidney biopsies performed on patients with conditions involving primarily glomeruli (diabetic glomerulopathy (23), Alport syndrome (37)) or causing more or less isolated changes of nephron structures outside the glomeruli (Balkan endemic nephropathy (15) and hemorrhagic fever with renal syndrome (19)). The aim was to explore the ultrastructural features of the PCBM. In patients with diabetic glomerulopathy, the PCBM was homogeneous, with a width ranging from normal to evidently increased (55-355 nm). In patients with Alport syndrome, the PCBM was homogeneous, with no substantial splitting or prominent thickening. Mean thickness varied between 80 (85-100) nm in children and 120 (46-250) nm in adults. Mean PCBM thickness in patients with Balkan endemic nephropathy was 209 (90-1270) nm. The thickened PCBM was also often split. In patients with hemorrhagic fever with renal syndrome, peritubular capillaries and medular vasa recta were generally extremely congested and focally ruptured, and their basal lamina showed prevailing thinning and focal discontinuities. PMID- 16316946 TI - Value of electron microscopy in kidney biopsy diagnosis. AB - Kidney biopsy reports given during 2003 were collected from the authors' pathology database. A total of 111 biopsies were performed. Five tumor samples were not studied with electron microscopy (EM). Of the remaining 106 biopsies, 85 were studied with EM. EM was not performed in 10/24 transplant biopsies, or in 11/82 cases of suspected primary kidney disease. The role of EM was evaluated by grouping the samples in 3 categories: (1) EM was essential for diagnosis, (2) EM contributed to the interpretation and cleared uncertainties, and (3) EM had no influence on the diagnostic process. In transplant biopsies EM influenced the final diagnosis in 86% of cases (category 2). In biopsies performed for primary kidney disease EM was essential for diagnosis in 18.3% clearly contributed in 53.5%, and had no influence on the final diagnosis in 28.2% of cases. The study suggests that the importance of EM has not decreased during the last few years. Because only about 25% of the EM reports did not have any influence on the diagnostic process, it is recommended that kidney biopsy protocols should include EM in all biopsy cases, or at least tissue should be reserved for EM studies of all cases. Because of the influence of EM on the diagnostic process the need for EM in pathology training should be emphasized. PMID- 16316947 TI - Electron microscopy in the diagnosis of BK-polyoma virus infection in the transplanted kidney. AB - BK polyomavirus has become an important etiologic agent responsible for significant morbidity in renal transplant recipients. This virus can be detected in transitional cells in the urine (decoy cells) using cytology, but correlation with allograft function status and histologic evidence of renal involvement is poor. Accurate diagnosis of BK polyomavirus infection requires a high index of suspicion and utilization of ancillary diagnostic techniques in many cases. Electron microscopy is very sensitive in depicting the presence of BK virions, but the finding of viral particles is not by itself diagnostic of BK interstitial nephritis. Management of patients with polyoma virus nephropathy is difficult since there is no specific antiviral therapy available at this time. PMID- 16316948 TI - Electron microscopy renders the diagnostic capabilities of cytopathology more precise: an approach to everyday practice. AB - Cytology is a powerful diagnostic tool but to make definitive diagnoses, the use of ancillary techniques is imperative. By combining immunohistochemistry (IHC) and electron microscopy (EM), cytologic diagnoses can be as precise as those of surgical pathology. In the authors' daily practice of cytopathology they use all ancillary techniques available to them: histochemistry, IHC, EM, flow cytometry, and molecular pathology. IHC is frequently used as an ancillary technique in their daily practice but EM is many times their technique of choice. By the use of EM the authors can make specific final diagnoses, make the diagnosis more definitive, narrow the differential diagnosis, or determine the origin of a neoplasm with unknown primary site. Specimens obtained by fine-needle aspiration as well as all body fluids are suitable for EM. The limiting factor is to obtain the appropriate material with the diagnostic cells for ultrastructural examination. The common diagnostic dilemmas in the everyday practice of cytology are the following: mesothelioma vs. adenocarcinoma, neuroendocrine differentiation or not, the distinction of melanoma from adenocarcinoma and sarcoma, hepatocellular carcinoma vs. adenocarcinoma, and the origin of adenocarcinomas of unknown primary. The authors discuss how they approach these diagnostic problems in their everyday practice and how they incorporate EM in solving them. PMID- 16316949 TI - Electron microscopic characteristics of beta2-microglobulin amyloid deposits in long-term haemodialysis. AB - The electron microscopic features of beta2-microglobulin amyloid, deposited in the synovial membrane, are presented and discussed. The patient, a 69-year-old woman underwent chronic hemodialysis for 3 years. Because of constant pain and destructive arthropathy, endoprosthesis of the hip joints were implanted. Extra- and intracellular filamentous-fibrillar amyloid deposits have been demonstrated in ultrathin sections. The extracellular amyloid deposits showed a loose, filamentous or fibrillar structure at the periphery and a dense central core. The loose, filamentous structure may represent an early stage of fresh, newly deposited beta2-microglobulin amyloid, while the condensed and fragmented amyloid filaments may be an advanced "mature" stage of amyloid deposition. PMID- 16316950 TI - Pulmonary blastomas of childhood: histologic, immunohistochemical, ultrastructural aspects and therapeutic considerations. AB - Pulmonary blastomas are rare neoplasms typically occurring in patients of pediatric age, clinically characterized by fever, respiratory distress, and radiologic findings of a pulmonary cystic and/or solid mass with partial or complete obliteration of emithorax. Their behavior is aggressive and outcome is poor due to frequent relapses and metastases. The histological, immunohistochemical, and ultrastructural aspects of a personal series of 6 cases of pulmonary blastoma are described and the differences between childhood and adult types are stressed. Due to the aggressiveness of these rare tumors, therapeutic management is quite difficult. The expression of the transmembrane tyrosin kinase receptor c-kit in all the solid cases of this series leads the authors to hypothesize new possible therapeutic implications for these tumors. PMID- 16316951 TI - Ultrastructure of lamellar bodies in congenital surfactant deficiency. AB - Congenital surfactant deficiency (CSD) is a newly identified neonatal lung disorder associated with a variety of molecular defects affecting surfactant synthesis and secretion in alveolar type II cells. The authors present ultrastructural findings of abnormal lamellar bodies in lung biopsies from 4 infants with CSD. All were term infants presenting shortly after birth with severe respiratory failure that was unresponsive to conventional therapy and all died within the first month of life. Lung biopsies were performed between 8 and 25 days of age. Biochemical and molecular studies in 2 unrelated male infants identified SP-B deficiency, one case with 121 ins 2 mutation and the second with a 209 + 4 A > G mutation. Light microscopy in both cases showed features of alveolar proteinosis. Ultrastructurally, alveolar type II cells lacked mature lamellar bodies, and their cytoplasm contained numerous pleomorphic inclusions with membranous and vesicular structures not seen in normal type II cells. The other 2 infants were a pair of siblings in whom molecular studies identified mutations in ABCA3 transporter gene. Light microscopy showed features of acinar dysplasia and desquamative interstitial pneumonitis. TEM studies revealed absence of mature lamellar bodies in type II cells and instead showed a mixture of cytoplasmic electron-dense inclusions with concentric membranes and distinctive electron dense aggregates. The ultrastructural changes in alveolar type II cells correlated well with specific gene defect. In SP-B deficiency, the absence of mature lamellar bodies is consistent with the postulated role for this protein in the formation of lamellar bodies. The lack of mature lamellar bodies in the ABCA3 gene mutations is due to the dysfunction of this endogenous lipid transporter that targets surfactant lipid moieties to the lamellar bodies. The findings demonstrate the importance of TEM studies of lung biopsies from infants with CSD as it is a critical adjunct in the diagnosis of neonatal lung disease and in defining the underlying cellular defects. PMID- 16316952 TI - Silent adenoma subtype 3 of the pituitary--immunohistochemical and ultrastructural classification: a review of 29 cases. AB - The silent adenoma subtype 3 (SAS-3) of undetermined cellular derivation is a seemingly nonfunctioning aggressive pituitary tumor with a high recurrence rate. At the time of diagnosis SAS-3s are macro- or giant adenomas particularly aggressive in young individuals, especially women. They are usually associated with mild hyperprolactinemia and are unremarkable by histology. Immunohistochemistry, demonstrating scattered immunoreactivity mostly for GH, PRL, TSH, and alpha-subunit, is not diagnostic. Presently, only TEM permits conclusive diagnosis. Ultrastructurally, the large polar adenoma cells contain abundant RER, masses of SER, extensive multipolar Golgi apparatus, and unevenly clustered mitochondria, displaced by RER and SER, which may show close spatial relationship to RER. Cell membranes often form plexiform interdigitations. Nuclear pleomorphism and nuclear inclusions are common. The 100- to 200-nm secretory granules accumulate heavily in cell processes, which is a hallmark of glycoprotein hormone cell differentiation. The endothelial cells may contain tubuloreticular inclusions. Complete surgical removal of the large often invasive tumors is difficult necessitating postoperative treatment. SAS-3 is sensitive to conventional radiation. Some tumors express somatostatin receptors and respond well to somatostatin analogues, offering long-term control in patients with residual tumor. Possible derivation of SAS-3 from rostral thyrotrophs, a cell type presently known in rodents is contemplated. PMID- 16316953 TI - Morphological and ultrastructural features of human islet grafts performed in diabetic nude mice. AB - Islet transplantation is a new therapeutic approach to type 1 diabetes mellitus. However, in several patients insulin levels are not restored and the glycemic control is inadequate. To clarify the cause of graft failure, the authors investigated with light and electron microscopy some human islet grafts before and after transplantation under the kidney capsule of streptozotocin-induced diabetic nude mice. In isolated islets, both pre- and post-transplantation, the endocrine component was scarcely represented, the beta/alpha cell ratio was reduced, and beta cells showed degenerative aspects such as apoptosis, immature secretory granules, and amylin fibrils deposition. The authors conclude that islet graft failure may be due to an insufficient beta cell mass related to their distress probably caused by anoxia and/or overstimulation. PMID- 16316954 TI - Ossifying fibromyxoid tumor: modified myoepithelial cell tumor? Report of three cases with immunohistochemical and electron microscopic studies. AB - Ossifying fibromyxoid tumors (OFMT) are rare soft tissue tumors of uncertain histogenesis and clinical behavior. Since Enzinger, Weiss, and Liang first described 59 examples in 1989 (Am Surg Pathol. 13:817-827), approximately 150 cases have been reported. Their clinicopathologic features are fairly well characterized and their histogenesis remains unknown. Three examples of soft tissue tumors with typical histopathologic characteristics of OFMT were studied: case 1, a 43-year-old female with a 2.5-cm tumor of the back; case 2, a 56-year old man with an 8-cm thigh mass; and case 3, an 81-year-old female with a 13.5-cm buttock tumor. For immunohistochemistry, formalin-fixed, paraffin-embedded tissue sections were stained with antibodies against cytokeratin, smooth muscle actin, desmin, vimentin, S-100 protein, EMA, and collagen type IV using standard ABC peroxidase methods. For electron microscopy, tissue samples fixed in EM-grade buffered formalin were processed according to routine methods. Immunohistochemistry showed that the tumor cells were positive for vimentin and S 100 protein in all 3 cases. Stains for collagen type IV revealed diffusely positive staining in the stroma with a tendency for stronger staining around the cell borders in 2 out of 3 cases. Desmin was positive in one and actin was positive in one other case. By electron microscopy, tumor cells were characterized by centrally located round to oval nuclei with varying amounts of cytoplasm containing scanty cytoplasmic organelles. There were rare profiles of rough-surfaced endoplasmic reticulum (RER) and rare mitochondria with areas of condensed intermediate filaments. No tonofilaments or actin filaments were present. There were multiple short web-like processes, some of which were attached to that of neighboring cells by primitive cell junctions. In all 3 cases, lesional cells showed external lamina (EL), which was abundant in case 1, forming redundant scrolls frequently. In case 2, EL was less prominent and incomplete, and interrupted portions of EL were present only along the periphery of cell columns or nests bordering the stroma. In case 3, which behaved as a malignant tumor, the tumor cells were less differentiated spindle cells with primitive cellular features, and EL was rarely found along the short span of tumor cell borders. In this study, tumor cells in OFMT were polygonal to stellate often with multiple short cytoplasmic processes. The tumor cells were found to form cell clusters attached by primitive intercellular junctions between cytoplasmic processes forming intercellular bridges. The cell borders facing the stroma around cell clusters tended to be flat and had incomplete EL, while no EL was present along the cell borders facing the inner aspect of cell clusters. These ultrastructural findings together with immunophenotypic expression of S-100 protein presented closer resemblance to those of modified myoepithelial cells in pleomorphic adenomas of salivary glands and skin appendages rather than peripheral nerve sheath tumors. The authors conclude that these findings render more support to the hypothesis of myoepithelial histogenesis of OFMT. They also conclude that ultrastructural study not only helps accurate diagnosis, but also may aid in predicting malignant behavior by the degree of deviation from the typical examples of OFMT. PMID- 16316955 TI - Cardiology patient page. Atrial flutter. PMID- 16316956 TI - Images in cardiovascular medicine. Ventricular septal defect complicating an acute myocardial infarction. PMID- 16316957 TI - Images in cardiovascular medicine. Ventricular septal rupture after a nonpenetrating chest trauma: findings from real-time three-dimensional echocardiography and cardiac magnetic resonance. PMID- 16316958 TI - Images in cardiovascular medicine. Giant blood cyst of the mitral valve: echocardiographic and intraoperative images. PMID- 16316959 TI - Letter regarding article by Pellikka et al, "Severe asymptomatic aortic stenosis". PMID- 16316960 TI - Apolipoprotein B versus non-high-density lipoprotein cholesterol: and the winner is... PMID- 16316961 TI - Weighing in before the fight: low-density lipoprotein cholesterol and non-high density lipoprotein cholesterol versus apolipoprotein B as the best predictor for coronary heart disease and the best measure of therapy. PMID- 16316962 TI - Association of blood pressure with genetic variation in WNK kinases in a white European population. PMID- 16316963 TI - Combined effects of systolic blood pressure and total cholesterol on cardiovascular disease risk. PMID- 16316964 TI - Non-high-density lipoprotein cholesterol and apolipoprotein B in the prediction of coronary heart disease in men. AB - BACKGROUND: Apolipoprotein B (apoB) plasma levels reflect the concentration of proatherogenic lipoproteins very low-density lipoprotein and low-density lipoprotein (LDL), whereas non-high-density lipoprotein cholesterol (non-HDL-C) levels reflect the concentration of cholesterol transported by these particles. METHODS AND RESULTS: The aim of our study was to compare apoB, non-HDL-C, LDL cholesterol (LDL-C), and other lipid markers as predictors of coronary heart disease (CHD) in a nested case-control study among 18 225 participants in the Health Professionals Follow-up Study. Among men who were free of diagnosed cardiovascular disease at the time of blood collection, 266 had nonfatal myocardial infarction or fatal CHD during 6 years of follow-up. Through the use of risk set sampling, control subjects were selected at a 2:1 ratio and matched with regard to age, date of blood collection, and smoking status. After adjustment for matching factors, the relative risk of CHD in the highest quintile compared with the lowest quintile was 2.76 (95% confidence interval [CI], 1.66 to 4.58) for non-HDL-C, 3.01 (95% CI, 1.81 to 5.00) for apoB, 1.81 (95% CI, 1.12 to 2.93) for LDL-C, 0.31 (95% CI, 0.18 to 0.52) for HDL-C, 2.41 (95% CI, 1.43 to 4.07) for triglycerides (all P trend <0.001), and 1.42 (95% CI, 0.86 to 2.32, P trend =0.19) for lipoprotein(a). When non-HDL-C and LDL-C were mutually adjusted, only non-HDL-C was predictive of CHD. When non-HDL-C and apoB were mutually adjusted, only apoB was predictive; the relative risk was 4.18 (95% CI, 1.30 to 13.49; P trend =0.02) for apoB compared with 0.70 (95% CI, 0.21 to 2.27; P trend =0.72) for non-HDL-C. Triglycerides added significant information to non-HDL-C but not to apoB for CHD risk prediction. CONCLUSIONS: Although non-HDL-C and apoB were both strong predictors of CHD in this male cohort, more so than LDL-C, the findings support the concept that the plasma concentration of atherogenic lipoprotein particles measured by apoB is more predictive in development of CHD than the cholesterol carried by these particles, measured by non-HDL-C. PMID- 16316965 TI - Critical role for monocyte chemoattractant protein-1 and macrophage inflammatory protein-1alpha in induction of experimental autoimmune myocarditis and effective anti-monocyte chemoattractant protein-1 gene therapy. AB - BACKGROUND: Autoimmune myocarditis is a principal cause of heart failure among young adults and is often a precursor of dilated cardiomyopathy. Monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1alpha (MIP 1alpha) are potent chemotactic factors for mononuclear cells. The inflammatory infiltrate observed in myocardial lesions of myocarditis consists of >70% mononuclear cells. To determine their critical role in the pathogenesis of myocarditis, we inhibited mononuclear cell activation and migration to see if it would affect disease severity and disease prevalence in experimental autoimmune myocarditis (EAM). METHODS AND RESULTS: In this report, we demonstrated that blockade of MCP-1 or MIP-1alpha with monoclonal antibodies significantly reduced severity of myocarditis in BALB/c mice immunized with cardiac myosin. Similar results were obtained when CCR2-/- and CCR5-/- mice were used. In CCR2-/- mice, not only disease severity but also disease prevalence was reduced. To further inhibit mononuclear cell activation and migration, we transfected the mice before inducing EAM with a dominant-negative inhibitor of MCP-1 gene (7ND). This transfection significantly reduced the disease severity, decreased mRNA expression levels, especially of the chemokines RANTES, MIP-2, IP-10, MCP-1, T cell activation gene 3, and eotaxin in the myocardium, and resulted in a reduction in cardiac myosin-induced interleukin-1 and interleukin-4 and in an increase in interferon-gamma and interleukin-10 cytokine production by splenocytes. CONCLUSIONS: Overall, these findings suggest that the chemokines MCP 1 and MIP-1alpha, acting through their receptors CCR2 and CCR5, are important in the induction of EAM and that inhibition of MCP-1 with 7ND gene transfection significantly reduced disease severity. This strategy may be a new feasible form of gene therapy against autoimmune myocarditis. PMID- 16316966 TI - Comparison of bare-metal stenting with minimally invasive bypass surgery for stenosis of the left anterior descending coronary artery: a 5-year follow-up. AB - BACKGROUND: Randomized trials comparing stenting with minimally invasive direct coronary artery bypass surgery in patients with isolated proximal left anterior descending lesions have shown a significantly higher reintervention rate for stenting and similar results for mortality and reinfarction at short-term follow up. Long-term follow-up data are sparse. METHODS AND RESULTS: Patients with isolated proximal left anterior descending stenosis were randomized to either surgery (n=110) or bare-metal stenting (n=110). At 5 years, follow-up data were obtained with respect to the primary end point of death, reinfarction, or repeated target vessel revascularization. Clinical symptoms were assessed by the Canadian Cardiovascular Society (CCS) classification. Follow-up information was completed for 216 patients (98.2%), and mean follow-up was 5.6+/-1.2 years. With respect to mortality (surgery, 12%; stenting, 10%; P=0.54) and reinfarctions (surgery, 7%; stenting, 5%; P=0.46), there were no differences between treatment strategies. The need for repeated target vessel revascularization was significantly higher after stenting (32%) compared with surgery (10%; P<0.001). Clinical symptoms improved significantly in both treatment groups compared with baseline; however, there was a favorable trend for surgery (stenting: CCS, 2.6+/ 0.9 to 0.5+/-0.8, P<0.001; surgery: CCS, 2.6+/-0.9 to 0.3+/-0.6, P<0.001; P=0.05, stenting versus surgery). CONCLUSIONS: At the 5-year follow-up, minimally invasive bypass surgery and bare-metal stenting showed similar results for the end points of mortality and reinfarctions. However, the reintervention rate is higher after stenting, and the relief in clinical symptoms is slightly better after surgery. PMID- 16316968 TI - Outcomes and associated risk factors for aortic valve replacement in 160 children: a competing-risks analysis. AB - BACKGROUND: We sought to define patient characteristics, outcomes, and associated risk factors after aortic valve replacement (AVR) in children. METHODS AND RESULTS: Clinical records from children undergoing AVR from 1974 to 2004 at our institution were reviewed. Competing-risks methodology determined the time related prevalence of 3 mutually exclusive end states: death, repeated replacement, and survival without subsequent AVR and their associated risk factors. Longitudinal echocardiographic data were analyzed by mixed linear regression models. Children (n=160) underwent 198 AVRs, with 33 having >1. Competing-risks analysis predicted that 10 years from the initial AVR, 19% had died without subsequent AVR, 34% underwent a second AVR, and 47% remained alive without replacement. Risk factors for death without a second AVR included lower weight (P<0.001) and younger age at AVR (P=0.04), performance of aortic arch reconstruction together with AVR (P=0.03), and nonautograft use (P=0.03). Risk factors for a second AVR included earlier operation year (P=0.04) and implantation of a bioprosthetic or homograft valve (P=0.004). Analysis of serial echocardiographic measurements showed that pulmonary autograft use was associated with slower progression of peak aortic gradient (P=0.002), smaller left ventricular dimension (P=0.04), and decreased prevalence of aortic regurgitation (P=0.04). CONCLUSIONS: Mortality and repeated valve replacement are common after initial AVR in children, especially in younger patients and those with bioprosthetic or homograft valves. Pulmonary autograft use is associated with decreased mortality, slower gradient progression, and smaller left ventricular dimension. PMID- 16316967 TI - Mitochondrial dysfunction and apoptosis underlie the pathogenic process in alpha B-crystallin desmin-related cardiomyopathy. AB - BACKGROUND: Mitochondria and sarcomeres have a well-defined architectural relation that partially depends on the integrity of the cytoskeletal network. An R120G missense mutation in the small heat shock protein alpha-B-crystallin (CryAB) causes desmin-related cardiomyopathy. Desmin-related cardiomyopathy is characterized by the formation of intracellular aggregates containing CryAB and desmin that are amyloid positive, and disease can be recapitulated in transgenic mice by cardiac-specific expression of the mutant protein. METHODS AND RESULTS: To understand the resultant pathology, we explored the acute effects of R120G expression both in vitro and in vivo. In vitro, transfection of adult cardiomyocytes with R120G-expressing adenovirus resulted in altered contractile mechanics. In vivo, as the cytoskeletal network is disturbed but before deficits in organ function can be detected, alterations in mitochondrial organization and architecture occur, leading to a reduction in the maximal rate of oxygen consumption with substrates that utilize complex I activity, alterations in the permeability transition pore, and compromised inner membrane potential. Apoptotic pathways are subsequently activated, which eventually results in cardiomyocyte death, dilation, and heart failure. CONCLUSIONS: Cardiac chaperone dysfunction acutely leads to altered cardiomyocyte mechanics, perturbations in mitochondrial sarcomere architecture, and deficits in mitochondrial function, which can result in activation of apoptosis and heart failure. PMID- 16316969 TI - Intravenous amiodarone for incessant tachyarrhythmias in children: a randomized, double-blind, antiarrhythmic drug trial. AB - BACKGROUND: Intravenous (IV) amiodarone has proven efficacy in adults. However, its use in children is based on limited retrospective data. METHODS AND RESULTS: A double-blind, randomized, multicenter, dose-response study of the safety and efficacy of IV amiodarone was conducted in 61 children (30 days to 14.9 years; median, 1.6 years). Children with incessant tachyarrhythmias (supraventricular arrhythmias [n=26], junctional ectopic tachycardia [JET, n=31], or ventricular arrhythmias [n=4]) were randomized to 1 of 3 dosing regimens (low, medium, or high: load plus 47-hour maintenance) with up to 5 open-label rescue doses. The primary efficacy end point was time to success. Of 229 patients screened, 61 were enrolled during 13 months by 27 of 48 centers in 7 countries. Median time to success was significantly related to dose (28.2, 2.6, and 2.1 hours for the low-, medium-, and high-dose groups, respectively; P=0.028). There was no significant association with dose for any arrhythmia subgroup, including JET, but the subgroups were too small for an accurate assessment. Adverse events (AEs) were common (87%), leading to withdrawal of 10 patients. There were 5 deaths in the 30 day follow-up period (2 possibly related to the study drug). Dose-related AEs included hypotension (36%), vomiting (20%), bradycardia (20%), atrioventricular block (15%) and nausea (10%). CONCLUSIONS: In children, the overall efficacy of IV amiodarone, as measured by time to success, was dose related but not significantly for any arrhythmia subgroup. AEs were common and appeared to be dose related. Although efficacious for critically ill patients, the dose-related risks of IV amiodarone should be taken into account when treating children with incessant arrhythmias. Prospective, placebo-controlled trials would be helpful in assessing antiarrhythmic drug efficacy in children, because their results may differ from retrospective series and adult studies. PMID- 16316970 TI - Multitargeted effects of statin-enhanced thrombolytic therapy for stroke with recombinant human tissue-type plasminogen activator in the rat. AB - BACKGROUND: Microvascular dysfunction posttreatment of stroke with recombinant human tissue-type plasminogen activator (rht-PA) constrains the therapeutic window to 3 hours. Statins (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) promote vascular thrombolysis and reduce the inflammation response. We therefore investigated the neuroprotective effects of a combination of atorvastatin and delayed rht-PA treatment in a rat model of embolic stroke. METHODS AND RESULTS: Rats subjected to embolic middle cerebral artery occlusion were treated with atorvastatin in combination with rht-PA 4 hours after stroke. Magnetic resonance imaging measurements revealed that combination treatment with atorvastatin and rht-PA blocked the expansion of the ischemic lesion, which improved neurological function compared with saline-treated rats. Real-time reverse transcription-polymerase chain reaction analysis of single endothelial cells isolated by laser-capture microdissection from brain tissue and immunostaining showed that combination treatment downregulated expression of tissue factor, von Willebrand factor, protease-activated receptor-1, intercellular adhesion molecule-1, and matrix metalloproteinase-9, which concomitantly reduced cerebral microvascular thrombosis and enhanced microvascular integrity. Combination treatment did not increase cerebrovascular endothelial nitric oxide synthase (eNOS) levels or eNOS activity, and inhibition of NOS activity with N-nitro-L-arginine methyl ester did not block the beneficial effects of combination treatment on stroke. Furthermore, combination treatment compared with thrombolytic monotherapy increased cerebral blood flow and reduced infarct volume in eNOS-null mice. CONCLUSIONS: These data demonstrate that combination treatment with atorvastatin and rht-PA exerts a neuroprotective effect when administered 4 hours after stroke and that the therapeutic benefits are likely attributed to its multitargeted effects on cerebrovascular patency and integrity. PMID- 16316971 TI - Exertional leg pain in patients with and without peripheral arterial disease. AB - BACKGROUND: Although exertional leg pain is a hallmark of peripheral arterial disease (PAD) and can occur in persons without PAD, symptom variation has received inadequate attention. METHODS AND RESULTS: Three cohort studies were combined for cross-sectional analysis. The San Diego Claudication Questionnaire assessed exertional leg pain. PAD was defined as ankle brachial index (ABI) < or =0.90 or history of lower-extremity revascularization. Of 3658 subjects, 3629 were analyzed after exclusions. Of these, 24.1% had PAD in 1 or both legs. There was a stepwise decrease in average ABI, from no pain to pain on exertion and rest, noncalf pain, atypical calf pain, and classic claudication (P=0.002). When stratified by PAD, this trend was no longer significant. Legs with ABIs >0.90 and revascularization had pain distributions intermediate between that of normal legs (ABI, 1.00 to 1.39) and legs with ABIs < or =0.90. Compared with normal legs, legs with low-normal (0.91 to 0.99) and high-normal (> or =1.40) ABIs had higher pain rates, suggesting borderline disease and vascular stiffness, respectively. Multivariable logistic regression models showed that ABI was a strong correlate of pain category throughout the ABI range. Independently of ABI, age, male sex, diabetes, smoking history, high body mass index, myocardial infarction, and previous revascularization were all significant correlates of exertional leg pain. CONCLUSIONS: No category of exertional leg pain was sufficiently sensitive or specific for routine PAD diagnosis. Legs with low-normal and high-normal ABIs appeared to have ischemic leg pain; thus, a "normal ABI" is likely to range from 1.00 to 1.39. In addition to ABI, several risk variables were independent correlates of exertional leg pain. PMID- 16316972 TI - Should patients with acute myocardial infraction be transferred to a tertiary center for primary angioplasty or receive it at qualified hospitals in community? The case for community hospital angioplasty. PMID- 16316973 TI - Long polyamines act as cofactors in PIP2 activation of inward rectifier potassium (Kir2.1) channels. AB - Phosphatidylinosital-4,5-bisphosphate (PIP2) acts as an essential factor regulating the activity of all Kir channels. In most Kir members, the dependence on PIP2 is modulated by other factors, such as protein kinases (in Kir1), G(betagamma) (in Kir3), and the sulfonylurea receptor (in Kir6). So far, however, no regulator has been identified in Kir2 channels. Here we show that polyamines, which cause inward rectification by selectively blocking outward current, also regulate the interaction of PIP2 with Kir2.1 channels to maintain channel availability. Using spermine and diamines as polyamine analogs, we demonstrate that both spontaneous and PIP2 antibody-induced rundown of Kir2.1 channels in excised inside-out patches was markedly slowed by long polyamines; in contrast, polyamines with shorter chain length were ineffective. In K188Q mutant channels, which have a low PIP2 affinity, application PIP2 (10 microM) was unable to activate channel activity in the absence of polyamines, but markedly activated channels in the presence of long diamines. Using neomycin as a measure of PIP2 affinity, we found that long polyamines were capable of strengthening either the wild type or K188Q channels' interaction with PIP2. The negatively charged D172 residue inside the transmembrane pore region was critical for the shift of channel-PIP2 binding affinity by long polyamines. Sustained pore block by polyamines was neither sufficient nor necessary for this effect. We conclude that long polyamines serve a dual role as both blockers and coactivators (with PIP2) of Kir2.1 channels. PMID- 16316974 TI - Electrostatics in the cytoplasmic pore produce intrinsic inward rectification in kir2.1 channels. AB - Inward rectifier K+ channels are important in regulating membrane excitability in many cell types. The physiological functions of these channels are related to their unique inward rectification, which has been attributed to voltage-dependent block. Here, we show that inward rectification can also be induced by neutral and positively charged residues at site 224 in the internal vestibule of tetrameric Kir2.1 channels. The order of extent of inward rectification is E224K mutant > E224G mutant > wild type in the absence of internal blockers. Mutating the glycines at the equivalent sites to lysines also rendered weak inward rectifier Kir1.1 channels more inwardly rectifying. Also, conjugating positively charged methanethiosulfonate to the cysteines at site 224 induced strong inward rectification, whereas negatively charged methanethiosulfonate alleviated inward rectification in the E224C mutant. These results suggest that charges at site 224 may control inward rectification in the Kir2.1 channel. In a D172N mutant, spermine interacting with E224 and E299 induced channel inhibition during depolarization but did not occlude the pore, further suggesting that a mechanism other than channel block is involved in the inward rectification of the Kir2.1 channel. In this and our previous studies we showed that the M2 bundle crossing and selectivity filter were not involved in the inward rectification induced by spermine interacting with E224 and E299. We propose that neutral and positively charged residues at site 224 increase a local energy barrier, which reduces K+ efflux more than K+ influx, thereby producing inward rectification. PMID- 16316975 TI - Separate ion pathways in a Cl-/H+ exchanger. AB - CLC-ec1 is a prokaryotic CLC-type Cl(-)/H+ exchange transporter. Little is known about the mechanism of H+ coupling to Cl-. A critical glutamate residue, E148, was previously shown to be required for Cl(-)/H+ exchange by mediating proton transfer between the protein and the extracellular solution. To test whether an analogous H+ acceptor exists near the intracellular side of the protein, we performed a mutagenesis scan of inward-facing carboxyl-bearing residues and identified E203 as the unique residue whose neutralization abolishes H+ coupling to Cl- transport. Glutamate at this position is strictly conserved in all known CLCs of the transporter subclass, while valine is always found here in CLC channels. The x-ray crystal structure of the E203Q mutant is similar to that of the wild-type protein. Cl- transport rate in E203Q is inhibited at neutral pH, and the double mutant, E148A/E203Q, shows maximal Cl- transport, independent of pH, as does the single mutant E148A. The results argue that substrate exchange by CLC-ec1 involves two separate but partially overlapping permeation pathways, one for Cl- and one for H+. These pathways are congruent from the protein's extracellular surface to E148, and they diverge beyond this point toward the intracellular side. This picture demands a transport mechanism fundamentally different from familiar alternating-access schemes. PMID- 16316976 TI - The glutamate transporter subtypes EAAT4 and EAATs 1-3 transport glutamate with dramatically different kinetics and voltage dependence but share a common uptake mechanism. AB - Here, we report the application of glutamate concentration jumps and voltage jumps to determine the kinetics of rapid reaction steps of excitatory amino acid transporter subtype 4 (EAAT4) with a 100-micros time resolution. EAAT4 was expressed in HEK293 cells, and the electrogenic transport and anion currents were measured using the patch-clamp method. At steady state, EAAT4 was activated by glutamate and Na+ with high affinities of 0.6 microM and 8.4 mM, respectively, and showed kinetics consistent with sequential binding of Na(+)-glutamate-Na+. The steady-state cycle time of EAAT4 was estimated to be >300 ms (at -90 mV). Applying step changes to the transmembrane potential, V(m), of EAAT4-expressing cells resulted in the generation of transient anion currents (decaying with a tau of approximately 15 ms), indicating inhibition of steady-state EAAT4 activity at negative voltages (<-40 mV) and activation at positive V(m) (>0 mV). A similar inhibitory effect at V(m) < 0 mV was seen when the electrogenic glutamate transport current was monitored, resulting in a bell-shaped I-V(m) curve. Jumping the glutamate concentration to 100 muM generated biphasic, saturable transient transport and anion currents (K(m) approximately 5 microM) that decayed within 100 ms, indicating the existence of two separate electrogenic reaction steps. The fast electrogenic reaction was assigned to Na+ binding to EAAT4, whereas the second reaction is most likely associated with glutamate translocation. Together, these results suggest that glutamate uptake of EAAT4 is based on the same molecular mechanism as transport by the subtypes EAATs 1-3, but that its kinetics and voltage dependence are dramatically different from the other subtypes. EAAT4 kinetics appear to be optimized for high affinity binding of glutamate, but not rapid turnover. Therefore, we propose that EAAT4 is a high-affinity/low-capacity transport system, supplementing low-affinity/high-capacity synaptic glutamate uptake by the other subtypes. PMID- 16316977 TI - A unique voltage sensor sensitizes the potassium channel AKT2 to phosphoregulation. AB - Among all voltage-gated K+ channels from the model plant Arabidopsis thaliana, the weakly rectifying K+ channel (K(weak) channel) AKT2 displays unique gating properties. AKT2 is exceptionally regulated by phosphorylation: when nonphosphorylated AKT2 behaves as an inward-rectifying potassium channel; phosphorylation of AKT2 abolishes inward rectification by shifting its activation threshold far positive (>200 mV) so that it closes only at voltages positive of +100 mV. In its phosphorylated form, AKT2 is thus locked in the open state in the entire physiological voltage range. To understand the molecular grounds of this unique gating behavior, we generated chimeras between AKT2 and the conventional inward-rectifying channel KAT1. The transfer of the pore from KAT1 to AKT2 altered the permeation properties of the channel. However, the gating properties were unaffected, suggesting that the pore region of AKT2 is not responsible for the unique K(weak) gating. Instead, a lysine residue in S4, highly conserved among all K(weak) channels but absent from other plant K+ channels, was pinpointed in a site-directed mutagenesis approach. Substitution of the lysine by serine or aspartate abolished the "open-lock" characteristic and converted AKT2 into an inward-rectifying channel. Interestingly, phosphoregulation of the mutant AKT2-K197S appeared to be similar to that of the K(in) channel KAT1: as suggested by mimicking the phosphorylated and dephosphorylated states, phosphorylation induced a shift of the activation threshold of AKT2-K197S by about +50 mV. We conclude that the lysine residue K197 sensitizes AKT2 to phosphoregulation. The phosphorylation-induced reduction of the activation energy in AKT2 is approximately 6 kT larger than in the K197S mutant. It is discussed that this hypersensitive response of AKT2 to phosphorylation equips a cell with the versatility to establish a potassium gradient and to make efficient use of it. PMID- 16316978 TI - Apocrine cysts of the breast: biomarkers, origin, enlargement, and relation with cancer phenotype. AB - Up to one-third of women aged 30-50 years have cysts in their breasts and are presumed to be at increased risk of developing breast cancer. Here we present an extensive proteomic and immunohistochemistry (IHC) study of breast apocrine cystic lesions aimed at generating specific biomarkers and elucidating the relationship, if existent, of apocrine cysts with cancer phenotype. To this end we compared the expression profiles of apocrine macrocysts obtained from mastectomies from high risk cancer patients with those of cancerous and non malignant mammary tissue biopsies collected from the same patients. We identified two biomarkers, 15-hydroxyprostaglandin dehydrogenase and 3-hydroxymethylglutaryl CoA reductase, that were expressed specifically by apocrine type I cysts as well as by apocrine metaplastic cells in type II microcysts, terminal ducts, and intraductal papillary lesions. No expression of these markers was observed in non malignant terminal ductal lobular units, type II flat cysts, stroma cells, or fat tissue as judged by IHC analysis of matched non-malignant tissue samples collected from 93 high risk patients enrolled in our cancer program. IHC analysis of the corresponding 93 primary tumors indicated that most apocrine changes have little intrinsic malignant potential, although some may progress to invasive apocrine cancer. None of the apocrine lesions examined, however, seemed to be a precursor of invasive ductal carcinomas, which accounted for 81% of the tumors analyzed. Our studies also provided some insight into the origin, development, and enlargement of apocrine cysts in mammary tissue. The successful identification of differentially expressed proteins that characterize specific steps in the progression from early benign lesions to apocrine cancer opens a window of opportunity for designing and testing new approaches for pharmacological intervention, not only in a therapeutic setting but also for chemoprevention, to inhibit cyst development as both 15-hydroxyprostaglandin dehydrogenase and 3-hydroxymethylglutaryl-CoA reductase are currently being targeted for chemoprevention strategies in various malignancies. PMID- 16316979 TI - Surfactant sodium lauryl sulfate enhances skin vaccination: molecular characterization via a novel technique using ultrafiltration capillaries and mass spectrometric proteomics. AB - The skin is a highly accessible organ and thus provides an attractive immune environment for cost-effective, simple, and needle-free delivery of vaccines and immunomodulators. In this study, we pretreated mouse skin with an anionic surfactant, sodium lauryl sulfate (SLS), for a short period of time (10 min) followed by epicutaneous vaccination with hen egg lysozyme antigen. We demonstrated for the first time that pretreatment of skin with surfactant SLS significantly enhances the production of antibody to hen egg lysozyme. Short term pretreatment with SLS disorganized the stratum corneum, extracted partial lamellar lipids, induced the maturation of Langerhans cells, and did not result in epidermis thickening. To reveal the mechanism underlying these changes, particularly at the molecular level, we used a novel proteomic technique using ultrafiltration capillaries and mass spectrometry to identify in vivo proteins/peptides secreted in the SLS-pretreated skin. Two secretory proteins, named as calcium-binding protein S100A9 and thymosin beta4, were identified by this novel technique. These two proteins thus may provide new insight into the enhancing effect of surfactants on skin vaccination. PMID- 16316980 TI - Comparative proteomic analysis provides new insights into chilling stress responses in rice. AB - Low temperature is one of the major abiotic stresses limiting the productivity and the geographical distribution of many important crops. To gain a better understanding of chilling stress responses in rice (Oryza sativa L. cv. Nipponbare), we carried out a comparative proteomic analysis. Three-week-old rice seedlings were treated at 6 degrees C for 6 or 24 h and then recovered for 24 h. Chilling treatment resulted in stress phenotypes of rolling leaves, increased relative electrolyte leakage, and decreased net photosynthetic rate. The temporal changes of total proteins in rice leaves were examined using two-dimensional electrophoresis. Among approximately 1,000 protein spots reproducibly detected on each gel, 31 protein spots were down-regulated, and 65 were up-regulated at least at one time point. Mass spectrometry analysis allowed the identification of 85 differentially expressed proteins, including well known and novel cold-responsive proteins. Several proteins showed enhanced degradation during chilling stress, especially the photosynthetic proteins such as Rubisco large subunit of which 19 fragments were detected. The identified proteins are involved in several processes, i.e. signal transduction, RNA processing, translation, protein processing, redox homeostasis, photosynthesis, photorespiration, and metabolisms of carbon, nitrogen, sulfur, and energy. Gene expression analysis of 44 different proteins by quantitative real time PCR showed that the mRNA level was not correlated well with the protein level. In conclusion, our study provides new insights into chilling stress responses in rice and demonstrates the advantages of proteomic analysis. PMID- 16316981 TI - Concanavalin A-captured glycoproteins in healthy human urine. AB - Both the urinary proteome and its glycoproteome can reflect human health status, and more directly, functions of kidney and urinary tracts. Because the high abundance protein albumin is not N-glycosylated, the urine N-glycoprotein enrichment procedure could deplete it, and urine proteome could thus provide a more detailed protein profile in addition to glycosylation information especially when albuminuria occurs in some kidney diseases. In terms of describing the details of urinary proteins, the urine glycoproteome is even a better choice than the proteome itself. Pooled urine samples from healthy volunteers were collected and acetone-precipitated for proteins. N-Linked glycoproteins enriched with concanavalin A affinity purification were separated and analyzed by SDS-PAGE reverse phase LC/MS/MS or two-dimensional LC/MS/MS. A total of 225 urinary proteins were identified based on two-hit criteria with reliability over 97% for each peptide. Among these proteins, 94 were identified in previous urine proteome works, 150 were annotated as glycoproteins in Swiss-Prot, and 43 were predicted as glycoproteins by NetNGlyc 1.0. A number of known biomarkers and disease related glycoproteins were identified. Because changes in protein quantity or the glycosylation status can lead to changes in the concanavalin A-captured glycoprotein profile, specific urine glycoproteome patterns might be observed for specific pathological conditions as multiplex urinary biomarkers. Knowledge of the urine glycoproteome is important in understanding kidney and body function. PMID- 16316982 TI - Distinct genes encode type II Topoisomerases for the nucleus and mitochondrion in the protozoan parasite Trypanosoma brucei. AB - Topoisomerases are essential for orderly nucleic acid metabolism and cell survival and are proven targets for clinically useful antimicrobial and anticancer drugs. Interest in the topologically intricate mitochondrial DNA (kinetoplast or kDNA) of Trypanosoma brucei brucei and related kinetoplastid protozoan parasites has led to many reports of type II topoisomerases that participate in kDNA metabolism (we term the T. brucei brucei gene TbTOP2mt). We have now identified and characterized two new genes for type II topoisomerases in T. brucei brucei, termed TbTOP2alpha and TbTOP2beta. Phylogenetically, they share a common node with other nuclear topoisomerases, clearly distinct from a clade that includes the previously reported kinetoplastid genes, all of which are homologs of TbTOP2mt. Southern blot analysis reveals the new genes are single copy and positioned approximately 1.7 kb apart. Cognate mRNAs are expressed in African trypanosomes, but only a single message is detected in Leishmania or Crithidia. TbTOP2alpha encodes an ATP-dependent topoisomerase that appears as a single approximately 170-kDa band on immunoblots and localizes to the nucleus; RNA interference leads to pleomorphic nuclear (but not kDNA) abnormalities and early growth arrest. The role of TbTOP2beta is unclear. Although transcribed in trypanosomes, TbTOP2beta is not detected by beta-specific antiserum, and RNAi silencing results in no obvious phenotype. These studies indicate that African trypanosomes and related kinetoplastid human pathogens are unusual in having independent topoisomerase II genes to service their nuclear and mitochondrial genomes, and they highlight TbTOP2alpha as a promising target for the development of much-needed new therapies. PMID- 16316983 TI - Oncogenic RAS induces accelerated transition through G2/M and promotes defects in the G2 DNA damage and mitotic spindle checkpoints. AB - Activating mutations of RAS are prevalent in thyroid follicular neoplasms, which commonly have chromosomal losses and gains. In thyroid cells, acute expression of HRAS(V12) increases the frequency of chromosomal abnormalities within one or two cell cycles, suggesting that RAS oncoproteins may interfere with cell cycle checkpoints required for maintenance of a stable genome. To explore this, PCCL3 thyroid cells with conditional expression of HRAS(V12) or HRAS(V12) effector mutants were presynchronized at the G(1)/S boundary, followed by activation of expression of RAS mutants and release from the cell cycle block. Expression of HRAS(V12) accelerated the G(2)/M phase by approximately 4 h and promoted bypass of the G(2) DNA damage and mitotic spindle checkpoints. Accelerated passage through G(2)/M and bypass of the G(2) DNA damage checkpoint, but not bypass of the mitotic spindle checkpoint, required activation of mitogen-activated protein kinase (MAPK). However, selective activation of the MAPK pathway was not sufficient to disrupt the G(2) DNA damage checkpoint, because cells arrested appropriately in G(2) despite conditional expression of HRAS(V12,S35) or BRAF(V600E). By contrast to the MAPK requirement for radiation-induced G(2) arrest, RAS-induced bypass of the mitotic spindle checkpoint was not prevented by pretreatment with MEK inhibitors. These data support a direct role for the MAPK pathway in control of G(2) progression and regulation of the G(2) DNA damage checkpoint. We propose that oncogenic RAS activation may predispose cells to genomic instability through both MAPK-dependent and independent pathways that affect critical checkpoints in G(2)/M. PMID- 16316984 TI - A conserved motif is prerequisite for the interaction of NAC with ribosomal protein L23 and nascent chains. AB - In eukaryotes, newly synthesized proteins interact co-translationally with a multitude of different ribosome-bound factors and chaperones including the conserved heterodimeric nascent polypeptide-associated complex (NAC) and a Hsp40/70-based chaperone system. These factors are thought to play an important role in protein folding and targeting, yet their specific ribosomal localizations, which are prerequisite for their functions, remain elusive. This study describes the ribosomal localization of NAC and the molecular details by which NAC is able to contact the ribosome and gain access to nascent polypeptides. We identified a conserved RRK(X)nKK ribosome binding motif within the beta-subunit of NAC that is essential for the entire NAC complex to attach to ribosomes and allow for its interaction with nascent polypeptide chains. The motif localizes within a potential loop region between two predicted alpha helices in the N terminus of betaNAC. This N-terminal betaNAC ribosome-binding domain was completely portable and sufficient to target an otherwise cytosolic protein to the ribosome. NAC modified with a UV-activatable cross-linker within its ribosome binding motif specifically cross-linked to L23 ribosomal protein family members at the exit site of the ribosome, providing the first evidence of NAC-L23 interaction in the context of the ribosome. Mutations of L23 reduced NAC ribosome binding in vivo and in vitro, whereas other eukaryotic ribosome associated factors such as the Hsp70/40 chaperones Ssb or Zuotin were unaffected. We conclude that NAC employs a conserved ribosome binding domain to position itself on the L23 ribosomal protein adjacent to the nascent polypeptide exit site. PMID- 16316985 TI - Crystal structure of the hypoxia-inducible form of 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase (PFKFB3): a possible new target for cancer therapy. AB - The hypoxia-inducible form of 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase (PFKFB3) plays a crucial role in the progression of cancerous cells by enabling their glycolytic pathways even under severe hypoxic conditions. To understand its structural architecture and to provide a molecular scaffold for the design of new cancer therapeutics, the crystal structure of the human form was determined. The structure at 2.1 A resolution shows that the overall folding and functional dimerization are very similar to those of the liver (PFKFB1) and testis (PFKFB4) forms, as expected from sequence homology. However, in this structure, the N-terminal regulatory domain is revealed for the first time among the PFKFB isoforms. With a beta-hairpin structure, the N terminus interacts with the 2-Pase domain to secure binding of fructose-6-phosphate to the active pocket, slowing down the release of fructose-6-phosphate from the phosphoenzyme intermediate product complex. The C-terminal regulatory domain is mostly disordered, leaving the active pocket of the fructose-2,6-bisphosphatase domain wide open. The active pocket of the 6-phosphofructo-2-kinase domain has a more rigid conformation, allowing independent bindings of substrates, fructose-6 phosphate and ATP, with higher affinities than other isoforms. Intriguingly, the structure shows an EDTA molecule bound to the fructose-6-phosphate site of the 6 phosphofructo-2-kinase active pocket despite its unfavorable liganding concentration, suggesting a high affinity. EDTA is not removable from the site with fructose-6-P alone but is with both ATP and fructose-6-P or with fructose 2,6-bisphosphate. This finding suggests that a molecule in which EDTA is covalently linked to ADP is a good starting molecule for the development of new cancer-therapeutic molecules. PMID- 16316986 TI - Core fucosylation regulates epidermal growth factor receptor-mediated intracellular signaling. AB - alpha1,6-Fucosyltransferase (Fut8) catalyzes the transfer of a fucose residue to N-linked oligosaccharides on glycoproteins via an alpha1,6-linkage to form core fucosylation in mammals. We recently found that disruption of the Fut8 gene induces severe growth retardation and early postnatal death. To investigate the molecular mechanism involved, we have established embryonic fibroblasts of Fut8+/+ and Fut8-/-, derived from wild-type and Fut8-null mice, respectively. Interestingly, the epidermal growth factor (EGF)-induced phosphorylation levels of the EGF receptor (EGFR) were substantially blocked in Fut8-/- cells, compared with Fut8+/+ cells, while there are no significant changes in the total activities of tyrosine phosphatase for phosphorylated EGFR between two cells. The inhibition of EGFR phosphorylation was completely restored by re-introduction of the Fut8 gene to Fut8-/- cells. Consistent with this, EGFR-mediated JNK or ERK activation was significantly suppressed in Fut8-/- cells. Finally, we found that the core fucosylation of N-glycans is required for the binding of the EGF to its receptor, whereas no effect was observed for the expression levels of EGFR on the cell surface. Collectively, these results strongly suggest that core fucosylation is essential for EGF receptor-mediated biological functions. PMID- 16316987 TI - Exopolysaccharides from Burkholderia cenocepacia inhibit neutrophil chemotaxis and scavenge reactive oxygen species. AB - Bacteria belonging to the Burkholderia cepacia complex are important opportunistic pathogens in compromised hosts, particularly patients with cystic fibrosis or chronic granulomatous disease. Isolates of B. cepacia complex may produce large amounts of exopolysaccharides (EPS) that endow the bacteria with a mucoid phenotype and appear to facilitate bacterial persistence during infection. We showed that EPS from a clinical B. cenocepacia isolate interfered with the function of human neutrophils in vitro; it inhibited chemotaxis and production of reactive oxygen species (ROS), both essential components of innate neutrophil mediated host defenses. These inhibitory effects were not due to cytotoxicity or interference with intracellular calcium signaling. EPS also inhibited enzymatic generation of ROS in cell-free systems, indicating that it scavenges these bactericidal products. B. cenocepacia EPS is structurally distinct from Pseudomonas aeruginosa alginate, yet they share the capacity to scavenge ROS and inhibit chemotaxis. These properties could explain why the two bacterial species resist clearance from the infected cystic fibrosis lung. PMID- 16316988 TI - Iodotyrosine deiodinase is the first mammalian member of the NADH oxidase/flavin reductase superfamily. AB - The enzyme responsible for iodide salvage in the thyroid, iodotyrosine deiodinase, was solubilized from porcine thyroid microsomes by limited proteolysis with trypsin. The resulting protein retained deiodinase activity and was purified using anion exchange, dye, and hydrophobic chromatography successively. Peptide sequencing of the final isolate identified the gene responsible for the deiodinase. The amino acid sequence of the porcine enzyme is highly homologous to corresponding genes in a variety of mammals including humans, and the mouse gene was expressed in human embryonic kidney 293 cells to confirm its identity. The amino acid sequence of the deiodinase suggests the presence of three domains. The N-terminal domain provides a membrane anchor. The intermediate domain contains the highest sequence variability and lacks homology to structural motifs available in the common databases. The C-terminal domain is highly conserved and resembles bacterial enzymes of the NADH oxidase/flavin reductase superfamily. A three-dimensional model of the deiodinase based on the coordinates of the minor nitroreductase of Escherichia coli indicates that a Cys common to all of the mammal sequences is located adjacent to bound FMN. However, the deiodinase is not structurally related to other known flavoproteins containing redox-active cysteines or the iodothyronine deiodinases containing an active site selenocysteine. PMID- 16316989 TI - The novel nucleoside analog R1479 (4'-azidocytidine) is a potent inhibitor of NS5B-dependent RNA synthesis and hepatitis C virus replication in cell culture. AB - Hepatitis C virus (HCV) polymerase activity is essential for HCV replication. Targeted screening of nucleoside analogs identified R1479 (4'-azidocytidine) as a specific inhibitor of HCV replication in the HCV subgenomic replicon system (IC(50) = 1.28 microM) with similar potency compared with 2'-C-methylcytidine (IC(50) = 1.13 microM). R1479 showed no effect on cell viability or proliferation of HCV replicon or Huh-7 cells at concentrations up to 2 mM. HCV replicon RNA could be fully cleared from replicon cells after prolonged incubation with R1479. The corresponding 5'-triphosphate derivative (R1479-TP) is a potent inhibitor of native HCV replicase isolated from replicon cells and of recombinant HCV polymerase (NS5B)-mediated RNA synthesis activity. R1479-TP inhibited RNA synthesis as a CTP-competitive inhibitor with a K(i) of 40 nM. On an HCV RNA derived template substrate (complementary internal ribosome entry site), R1479-TP showed similar potency of NS5B inhibition compared with 3'-dCTP. R1479-TP was incorporated into nascent RNA by HCV polymerase and reduced further elongation with similar efficiency compared with 3'-dCTP under the reaction conditions. The S282T point mutation in the coding sequence of NS5B confers resistance to inhibition by 2'-C-MeATP and other 2'-methyl-nucleotides. In contrast, the S282T mutation did not confer cross-resistance to R1479. PMID- 16316990 TI - Phosphoprotein Crh-Ser46-P displays altered binding to CcpA to effect carbon catabolite regulation. AB - In Gram-positive bacteria, the catabolite control protein A (CcpA) functions as the master transcriptional regulator of carbon catabolite repression/regulation (CCR). To effect CCR, CcpA binds a phosphoprotein, either HPr-Ser46-P or Crh Ser46-P. Although Crh and histidine-containing protein (HPr) are structurally homologous, CcpA binds Crh-Ser46-P more weakly than HPr-Ser46-P. Moreover, Crh can form domain-swapped dimers, which have been hypothesized to be functionally relevant in CCR. To understand the molecular mechanism of Crh-Ser46-P regulation of CCR, we determined the structure of a CcpA-(Crh-Ser46-P)-DNA complex. The structure reveals that Crh-Ser46-P does not bind CcpA as a dimer but rather interacts with CcpA as a monomer in a manner similar to that of HPr-Ser46-P. The reduced affinity of Crh-Ser46-P for CcpA as compared with that of HPr-Ser46 P is explained by weaker Crh-Ser46-P interactions in its contact region I to CcpA, which causes this region to shift away from CcpA. Nonetheless, the interface between CcpA and helix alpha 2 of the second contact region (contact region II) of Crh-Ser46-P is maintained. This latter finding demonstrates that this contact region is necessary and sufficient to throw the allosteric switch to activate cre binding by CcpA. PMID- 16316991 TI - Mechanism of proteasomal degradation of inositol trisphosphate receptors in CHO K1 cells. AB - myo-Inositol 1,4,5-trisphosphate receptor (IP3R) degradation occurs in response to carbachol (Cch) stimulation of CHO-K1 cells. The response was mediated by endogenous muscarinic receptors and was blocked by atropine or proteasomal inhibitors. We have used these cells to identify the sites of ubiquitination on IP3Rs and study the role of Ca2+ and substrate recognition properties of the degradation system using exogenously expressed IP3R constructs. Employing caspase 3 for IP3R cleavage, we show that Cch promotes polyubiquitination in the N terminal domain and monoubiquitination in the C-terminal domain. The addition of extracellular Ca2+ to Ca2+-depleted Chinese hamster ovary (CHO) cells initiates IP3R degradation provided Cch is present. This effect is inhibited by thapsigargin. The data suggest that both a sustained elevation of IP3 and a minimal content of Ca2+ in the endoplasmic reticulum lumen is required to initiate IP3R degradation. Transient transfection of IP3R constructs into CHO cells indicated the selective degradation of only the SI+ splice variant of the type I IP3R. This was also the splice form present endogenously in these cells. A pore-defective, nonfunctional SI+ IP3R mutant (D2550A) was also degraded in Cch stimulated cells. The Cch-mediated response in CHO cells provides a convenient model system to further analyze the Ca2+ dependence and structural requirements of the IP3R proteasomal degradation pathway. PMID- 16316992 TI - Proteomic analysis of beta1-adrenergic receptor interactions with PDZ scaffold proteins. AB - Many G protein-coupled receptors possess carboxyl-terminal motifs ideal for interaction with PDZ scaffold proteins, which can control receptor trafficking and signaling in a cell-specific manner. To gain a panoramic view of beta1 adrenergic receptor (beta AR) interactions with PDZ scaffolds, the beta1AR carboxyl terminus was screened against a newly developed proteomic array of PDZ domains. These screens confirmed beta1AR associations with several previously identified PDZ partners, such as PSD-95, MAGI-2, GIPC, and CAL. Moreover, two novel beta1AR-interacting proteins, SAP97 and MAGI-3, were also identified. The beta1AR carboxyl terminus was found to bind specifically to the first PDZ domain of MAGI-3, with the last four amino acids (E-S-K-V) of beta1AR being the key determinants of the interaction. Full-length beta1AR robustly associated with full-length MAGI-3 in cells, and this association was abolished by mutation of the beta1AR terminal valine residue to alanine (V477A), as determined by co immunoprecipitation experiments and immunofluorescence co-localization studies. MAGI-3 co-expression with beta1AR profoundly impaired beta1AR-mediated ERK1/2 activation but had no apparent effect on beta1AR-mediated cyclic AMP generation or agonist-promoted beta1AR internalization. These findings revealed that the interaction of MAGI-3 with beta1AR can selectively regulate specific aspects of receptor signaling. Moreover, the screens of the PDZ domain proteomic array provide a comprehensive view of beta1AR interactions with PDZ scaffolds, thereby shedding light on the molecular mechanisms by which beta1 AR signaling and trafficking can be regulated in a cell-specific manner. PMID- 16316993 TI - Quantitative proteomic analysis of myc-induced apoptosis: a direct role for Myc induction of the mitochondrial chloride ion channel, mtCLIC/CLIC4. AB - Myc is a key regulatory protein in higher eukaryotes controlling important cellular functions such as proliferation, differentiation, and apoptosis. Myc is profoundly involved in the genesis of many human and animal cancers, and the abrogation of Myc-induced apoptosis is a critical event in cancer progression. Because the mechanisms that mediate Myc-induced apoptosis are largely unknown, we analyzed protein expression during Myc-induced apoptosis using an isotope-coded affinity tag quantitative proteomics approach and identified that a proapoptotic mitochondrial chloride ion channel, mtCLIC/CLIC4, is induced by Myc. Myc binds to the mtCLIC gene promoter and activates its transcription. Suppression of mtCLIC expression by RNA interference inhibited Myc-induced apoptosis in response to different stress conditions and abolished the cooperative induction of apoptosis by Myc and Bax. We also found that Myc reduces the expression of Bcl-2 and Bcl-xL and that the apoptosis-inducing stimuli up-regulate Bax expression. These results suggest that up-regulation of mtCLIC, together with a reduction in Bcl-2 and Bcl xL, sensitizes Myc-expressing cells to the proapoptotic action of Bax. PMID- 16316994 TI - Golgi localization determinants in ArfGAP1 and in new tissue-specific ArfGAP1 isoforms. AB - The Arf1-directed GTPase-activating protein ArfGAP1 is a Golgi-localized protein that controls the dynamics of the COPI coat of carriers that mediate transport in the endoplasmic reticulum-Golgi shuttle. Previously the interaction of ArfGAP1 with the Golgi was allocated to a portion of the non-catalytic, carboxyl part of the protein, but the mechanism of this interaction has not been established. In this study we identify a short stretch in the non-catalytic part of ArfGAP1 (residues 204-214) in which several hydrophobic residues contribute to Golgi localization. Even single alanine replacement of two of these residues (Leu-207 and Trp-211) strongly diminished Golgi localization. Mutations in the hydrophobic residues also diminished the in vitro activity of ArfGAP1 on Arf1 bound to Golgi membranes. The stretch containing the hydrophobic residues was recently shown to mediate the binding of ArfGAP1 to loosely packed lipids of highly curved liposomes (Bigay, J., Casella, J. F., Drin, G., Mesmin, B., and Antonny, B. (2005) EMBO J. 24, 2244-2253). Whereas short fragments containing the hydrophobic stretch were not Golgi-localized, a proximal 10-residue in-frame insertion that is present in new ArfGAP1 isoforms that we identified in brain and heart tissues could confer Golgi localization on these fragments. This localization was abrogated by alanine replacement of residues Phe-240 or Trp-241 of the insertion sequence but not by their replacement with leucines. Our findings indicate that ArfGAP1 interacts with the Golgi through multiple hydrophobic motifs and that alternative modes of interaction may exist in tissue-specific ArfGAP1 isoforms. PMID- 16316995 TI - IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. AB - Engagement of the high affinity receptor for IgE (FcepsilonRI) on mast cells results in the production and secretion of sphingosine 1-phosphate (S1P), a lipid metabolite present in the lungs of allergen-challenged asthmatics. Herein we report that two isoforms of sphingosine kinase (SphK1 and SphK2) are expressed and activated upon FcepsilonRI engagement of bone marrow-derived mast cells (BMMC). Fyn kinase is required for FcepsilonRI coupling to SphK1 and -2 and for subsequent S1P production. Normal activation of SphK1 and -2 was restored by expression of wild type Fyn but only partly with a kinase-defective Fyn, indicating that induction of SphK1 and SphK2 depended on both catalytic and noncatalytic properties of Fyn. Downstream of Fyn, the requirements for SphK1 activation differed from that of SphK2. Whereas SphK1 was considerably dependent on the adapter Grb2-associated binder 2 and phosphatidylinositol 3-OH kinase, SphK2 showed minimal dependence on these molecules. Fyn-deficient BMMC were defective in chemotaxis and, as previously reported, in degranulation. These functional responses were partly reconstituted by the addition of exogenous S1P to FcepsilonRI-stimulated cells. Taken together with our previous study, which demonstrated delayed SphK activation in Lyn-deficient BMMC, we propose a cooperative role between Fyn and Lyn kinases in the activation of SphKs, which contributes to mast cell responses. PMID- 16316996 TI - The RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. AB - Epac-1 and -2 (exchange proteins directly activated by cyclic AMP) are guanine nucleotide exchange factors for the GTPases Rap1 and -2. Epac2 but not Epac1 was found to possess a RA (Ras association) domain similar to that found in the Ras effector Ral-GDS. This domain specifically bound Ras-GTP, enabling oncogenic Ras to translocate Epac2 from the cytosol to the plasma membrane. Consequently, a small pool of plasma membrane-bound Rap1 was activated at the expense of bulk Rap1 located on intracellular organelles. Whereas translocation of Epac2 was not mimicked by challenge with epidermal growth factor alone, costimulation with forskolin, prostaglandin E2, or an Epac-selective cyclic AMP analog-induced rapid relocation of GFP-Epac2 but not -Epac1 to the plasma membrane in a Ras-dependent manner. Deletion of the cyclic AMP-binding domain overcame the need for nucleotide, suggesting that this domain normally masked the RA domain in the resting GEF. Thus, Epac2 can respond to costimulation by agonists that jointly elevate Ras-GTP and cyclic AMP levels, activating a specific pool of Rap1 at the plasma membrane. Therefore, despite its previous description as a Ras antagonist or independently functioning GTPase, Rap1/Krev-1 may additionally act downstream of Ras in cells that express the cyclic AMP-regulated GEF, Epac2. PMID- 16316997 TI - Sustained activation of phosphatidylinositol 3-kinase/Akt/nuclear factor kappaB signaling mediates G protein-coupled delta-opioid receptor gene expression. AB - Expression of the delta-opioid receptor gene (dor) is tightly controlled during neuronal differentiation and developmental stages. Such distinct temporal and spatial expression of dor during development suggests a role for the delta-opioid receptor in early developmental events. However, little is known about intracellular signaling pathways that control dor expression. A well established cell line model for the study of gene expression during neuronal differentiation is the rat adrenal pheochromocytoma PC12 cell line. Here we found that the constitutively activated TrkA/phosphatidylinositol 3-kinase/Akt (protein kinase B)/NF-kappaB survival cascade mediates dor expression during nerve growth factor (NGF)-induced differentiation of PC12h cells. Biochemical experiments showed that constitutive phosphorylation of Akt and IkappaBalpha correlates with NGF-induced dor expression. Overexpression of the transcriptional activator NF-kappaB/p65 increased dor promoter activity. Overexpression of the NF-kappaB signaling super inhibitor mutant IkappaBalpha (S32A/S36A) abolished the effect of p65 and blocked NGF-induced activation of NF-kappaB signaling, resulting in a significant reduction in dor promoter activity. Treatment with SN50, an NF-kappaB-specific nuclear translocation peptide inhibitor, inhibited the translocation of NF kappaB, resulting in a reduction of dor mRNA. The gel shift assay supported the fact that there exists an NF-kappaB-binding site on the dor promoter. RNA interference experiments using NF-kappaB/p65 small interfering RNA confirmed that NF-kappaB signaling is required for dor expression. Our findings not only provide a new mechanistic explanation for NGF-induced dor expression but also shed some light on the molecular mechanism of the temporal and spatial expression of dor and the roles of the delta-opioid receptor during neuronal differentiation. PMID- 16316998 TI - Activation of T cell calcium influx by the second messenger ADP-ribose. AB - Stimulation of Jurkat T cells by high concentrations of concanavalin A (ConA) induced an elevation of the endogenous adenosine diphosphoribose (ADPR) concentration and an inward current significantly different from the Ca2+ release activated Ca2+ current (I(CRAC)). Electrophysiological characterization and activation of a similar current by infusion of ADPR indicated that the ConA induced current is carried by TRPM2. Expression of TRPM2 in the plasma membrane of Jurkat T cells was demonstrated by reverse transcription-PCR, Western blot, and immunofluorescence. Inhibition of ADPR formation reduced ConA-mediated, but not store-operated, Ca2+ entry and prevented ConA-induced cell death of Jurkat cells. Moreover, gene silencing of TRPM2 abolished the ADPR- and ConA-mediated inward current. Thus, ADPR is a novel second messenger significantly involved in ConA-mediated cell death in T cells. PMID- 16316999 TI - Induction of apoptosis by the Ste20-like kinase SLK, a germinal center kinase that activates apoptosis signal-regulating kinase and p38. AB - Expression and activity of the germinal center kinase, Ste20-like kinase (SLK), are increased during kidney development and recovery from ischemic acute renal failure. In this study, we characterize the activation and functional role of SLK. SLK underwent dimerization via the C-terminal domain, and dimerization enhanced SLK activity. In contrast, the C-terminal domain of SLK did not dimerize with a related kinase, Mst1, and did not affect Mst1 activity. Phosphorylation/dephosphorylation of SLK were not associated with changes in kinase activity. SLK induced phosphorylation of apoptosis signal-regulating kinase-1 (ASK1) and increased ASK1 activity, indicating that ASK1 is a substrate of SLK. Moreover, SLK stimulated phosphorylation of p38 mitogen-activated protein kinase via ASK1, but not c-Jun N-terminal kinase nor extracellular signal regulated kinase. Chemical anoxia and recovery during re-exposure to glucose (ischemia-reperfusion injury in cell culture) stimulated SLK activity. Overexpression of SLK enhanced anoxia/recovery-induced apoptosis, release of cytochrome c, and activities of caspase-8 and -9, and apoptosis was reduced significantly with p38 and caspase-9 inhibitors. Induction of the endoplasmic reticulum stress response by anoxia/recovery or tunicamycin (monitored by induction of Bip or Grp94 expression, phosphorylation of eukaryotic translation initiation factor 2alpha subunit, expression of CHOP, and activation of caspase 12) was attenuated in cells that overexpress SLK. Thus, SLK is an anoxia/recovery dependent kinase that is activated via homodimerization and that signals via ASK1 and p38 to promote apoptosis. Attenuation of the protective aspects of the endoplasmic reticulum stress response by SLK may contribute to its proapoptotic effect. PMID- 16317000 TI - Crystal structure of a viral FLIP: insights into FLIP-mediated inhibition of death receptor signaling. AB - Death receptor signaling is initiated by the assembly of the death-inducing signaling complex, which culminates in the activation of the initiator caspase, either caspase-8 or caspase-10. A family of viral and cellular proteins, known as FLIP, plays an essential role in the regulation of death receptor signaling. Viral FLIP (v-FLIP) and short cellular FLIP (c-FLIPS) inhibit apoptosis by interfering with death receptor signaling. The structure and mechanisms of v-FLIP and c-FLIPS remain largely unknown. Here we report a high resolution crystal structure of MC159, a v-FLIP derived from the molluscum contagiosum virus, which is a member of the human poxvirus family. Unexpectedly, the two tandem death effector domains (DEDs) of MC159 rigidly associate with each other through a hydrophobic interface. Structure-based sequence analysis suggests that this interface is conserved in the tandem DEDs from other v-FLIP, c-FLIPS, and caspase 8 and -10. Strikingly, the overall packing arrangement between the two DEDs of MC159 resembles that between the caspase recruitment domains of Apaf-1 and caspase-9. In addition, each DED of MC159 contains a highly conserved binding motif on the surface, to which loss-of-function mutations in MC159 map. These observations, in conjunction with published evidence, reveal significant insights into the function of v-FLIP and suggest a mechanism by which v-FLIP and c-FLIPS inhibit death receptor signaling. PMID- 16317001 TI - Structure of the N-terminal calcium sensor domain of centrin reveals the biochemical basis for domain-specific function. AB - Centrin is an essential component of microtubule-organizing centers in organisms ranging from algae and yeast to humans. It is an EF-hand calcium-binding protein with homology to calmodulin but distinct calcium binding properties. In a previously proposed model, the C-terminal domain of centrin serves as a constitutive anchor to target proteins, and the N-terminal domain serves as the sensor of calcium signals. The three-dimensional structure of the N-terminal domain of Chlamydomonas rheinhardtii centrin has been determined in the presence of calcium by solution NMR spectroscopy. The domain is found to occupy an open conformation typical of EF-hand calcium sensors. Comparison of the N- and C terminal domains of centrin reveals a structural and biochemical basis for the domain specificity of interactions with its cellular targets and the distinct nature of centrin relative to other EF-hand proteins. An NMR titration of the centrin N-terminal domain with a fragment of the known centrin target Sfi1 reveals binding of the peptide to a discrete site on the protein, which supports the proposal that the N-terminal domain serves as a calcium sensor in centrin. PMID- 16317002 TI - The presumptive phosphatidylserine receptor is dispensable for innate anti inflammatory recognition and clearance of apoptotic cells. AB - The role of the presumptive phosphatidylserine receptor (PSR) in the recognition and engulfment of apoptotic cells, and the antiinflammatory response they exert, has been of great interest. Genetic deficiency of PSR in the mouse is lethal perinatally, and results to date have been ambiguous with regard to the phagocytic and inflammatory phenotypes associated with that deficiency. Recently, we found that the specific functional recognition of apoptotic cells is a ubiquitous property of virtually all cell types, including mouse embryo fibroblasts, and reflects an innate immunity that discriminates live from effete cells. Taking advantage of this property of fibroblasts, we generated, PSR(+/+), PSR(+/-), and PSR(-/-) fibroblast cell lines to examine definitively the involvement of PSR in apoptotic recognition and inflammatory modulation. Our data demonstrate that PSR-deficient cells are fully competent to recognize, engulf, and respond to apoptotic cells. Signal transduction in the responder cells, including the activation of Akt and Rac1, is unimpaired in the absence of PSR. We confirm as well that PSR is localized predominantly to the nucleus. However, it does not play a role in pro-inflammatory transcription or in the anti inflammatory modulation of that transcriptional response triggered by apoptotic cells. We conclude that PSR is not involved generally in either specific innate recognition or engulfment of apoptotic cells. PMID- 16317003 TI - Endoplasmic reticulum stress induces apoptosis by an apoptosome-dependent but caspase 12-independent mechanism. AB - The endoplasmic reticulum (ER) is the cellular site of polypeptide folding and modification. When these processes are hampered, an unfolded protein response (UPR) is activated. If the damage is too broad, the mammalian UPR launches the apoptotic program. As a consequence, mobilization of ER calcium stores sensitizes mitochondria to direct proapoptotic stimuli. We make use of a mouse Apaf1 deficient cell system of proneural origin to understand the roles played in this context by the apoptosome, the most studied apoptotic machinery along the mitochondrial pathway of death. We show here that in the absence of the apoptosome ER stress induces cytochrome c release from the mitochondria but that apoptosis cannot occur. Under these circumstances, Grp78/BiP and GADD153/CHOP, both hallmarks of UPR, are canonically up-regulated, and calcium is properly released from ER stores. We also demonstrate that caspase 12, a protease until now believed to play a central role in the initiation of ER stress-induced cell death in the mouse system, is dispensable for the mitochondrial pathway of death to take place. PMID- 16317004 TI - Involvement of endoplasmic reticulum stress in hereditary tyrosinemia type I. AB - Hereditary tyrosinemia type I (HTI) is the most severe disease of the tyrosine degradation pathway. HTI is caused by a deficiency of fumarylacetoacetate hydrolase (FAH), the enzyme responsible for the hydrolysis of fumarylacetoacetate (FAA). As a result, there is an accumulation of metabolites such as maleylacetoacetate, succinylacetone, and FAA. The latter was shown to display mutagenic, cytostatic, and apoptogenic activities and to cause chromosomal instability. Herein, we demonstrate that FAA also causes a cellular insult leading to the endoplasmic reticulum (ER) stress signaling. Treatment of V79 Chinese hamster lung cells with an apoptogenic dose of FAA (100 mum) causes an early induction of the ER resident chaperone GRP78/BiP and a simultaneous phosphorylation of the eIF2alpha. FAA treatment also causes a subsequent induction of the proapoptotic CHOP (CEBP homologous protein) transcription factor as well as a late activation of caspase-12. Data obtained from fah(-/-) mice taken off the therapeutic 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3 cyclohexanedione drug are similar. However, in this mouse model, there is also an increase in proteasome activity indicative of ER-associated degradation. This difference observed between the two models may be due to the fact that the murine model measures the effects of all metabolites accumulating in hereditary tyrosinemia type I as opposed to the cellular model that only measures the effects of exogenous FAA. PMID- 16317005 TI - The Schizosaccharomyces pombe replication inhibitor Spd1 regulates ribonucleotide reductase activity and dNTPs by binding to the large Cdc22 subunit. AB - Ribonucleotide reductase (RNR) is an essential enzyme that provides the cell with a balanced supply of deoxyribonucleoside triphosphates for DNA replication and repair. Mutations that affect the regulation of RNR in yeast and mammalian cells can lead to genetic abnormalities and cell death. We have expressed and purified the components of the RNR system in fission yeast, the large subunit Cdc22p, the small subunit Suc22p, and the replication inhibitor Spd1p. It was proposed (Liu, C., Powell, K. A., Mundt, K., Wu, L., Carr, A. M., and Caspari, T. (2003) Genes Dev. 17, 1130-1140) that Spd1 is an RNR inhibitor, acting by anchoring the Suc22p inside the nucleus during G1 phase. Using in vitro assays with highly purified proteins we have demonstrated that Spd1 indeed is a very efficient inhibitor of fission yeast RNR, but acting on Cdc22p. Furthermore, biosensor technique showed that Spd1p binds to the Cdc22p with a KD of 2.4 microM, whereas the affinity to Suc22p is negligible. Therefore, Spd1p inhibits fission yeast RNR activity by interacting with the Cdc22p. Similar to the situation in budding yeast, logarithmically growing fission yeast increases the dNTP pools 2-fold after 3 h of incubation in the UV mimetic 4-nitroquinoline-N-oxide. This increase is smaller than the increase observed in budding yeast but of the same order as the dNTP pool increase when synchronous Schizosaccharomyces pombe cdc10 cells are going from G1 to S-phase. PMID- 16317006 TI - Temperature-dependent biosynthesis of 2-thioribothymidine of Thermus thermophilus tRNA. AB - 2-Thioribothymidine (s(2)T) is a modified nucleoside of U, specifically found at position 54 of tRNAs from extreme thermophilic microorganisms. The function of the 2-thiocarbonyl group of s(2)T54 is thermostabilization of the three dimensional structure of tRNA; however, its biosynthesis has not been clarified until now. Using an in vivo tRNA labeling experiment, we demonstrate that the sulfur atom of s(2)T in tRNA is derived from cysteine or sulfate. We attempted to reconstitute 2-thiolation of s(2)T in vitro, using a cell extract of Thermus thermophilus. Specific 2-thiolation of ribothymidine, at position 54, was observed in vitro, in the presence of ATP. Using this assay, we found a strong temperature dependence of the 2-thiolation reaction in vitro as well as expression of 2-thiolation enzymes in vivo. These results suggest that the variable content of s(2)T in vivo at different temperatures may be explained by the above characteristics of the enzymes responsible for the 2-thiolation reaction. Furthermore, we found that another posttranscriptionally modified nucleoside, 1-methyladenosine at position 58, is required for the efficient 2 thiolation of ribothymidine 54 both in vivo and in vitro. PMID- 16317007 TI - Stathmin strongly increases the minus end catastrophe frequency and induces rapid treadmilling of bovine brain microtubules at steady state in vitro. AB - Stathmin is a ubiquitous microtubule destabilizing protein that is believed to play an important role linking cell signaling to the regulation of microtubule dynamics. Here we show that stathmin strongly destabilizes microtubule minus ends in vitro at steady state, conditions in which the soluble tubulin and microtubule levels remain constant. Stathmin increased the minus end catastrophe frequency approximately 13-fold at a stathmin:tubulin molar ratio of 1:5. Stathmin steady state catastrophe-promoting activity was considerably stronger at the minus ends than at the plus ends. Consistent with its ability to destabilize minus ends, stathmin strongly increased the treadmilling rate of bovine brain microtubules. By immunofluorescence microscopy, we also found that stathmin binds to purified microtubules along their lengths in vitro. Co-sedimentation of purified microtubules polymerized in the presence of a 1:5 initial molar ratio of stathmin to tubulin yielded a binding stoichiometry of 1 mol of stathmin per approximately 14.7 mol of tubulin in the microtubules. The results firmly establish that stathmin can increase the steady-state catastrophe frequency by a direct action on microtubules, and furthermore, they indicate that an important regulatory action of stathmin in cells may be to destabilize microtubule minus ends. PMID- 16317008 TI - Organization of calcium channel beta1a subunits in triad junctions in skeletal muscle. AB - In skeletal muscle, dihydropyridine receptors (DHPRs) in the plasma membrane interact with the type 1 ryanodine receptor (RyR1) at junctions with the sarcoplasmic reticulum. This interaction organizes junctional DHPRs into groups of four termed tetrads. In addition to the principle alpha1S subunit, the beta1a subunit of the DHPR is also important for the interaction with RyR1. To probe this interaction, we measured fluorescence resonance energy transfer (FRET) of beta1a subunits labeled with cyan fluorescent protein (CFP) and/or yellow fluorescent protein (YFP). Expressed in dysgenic (alpha1S-null) myotubes, YFP beta1a-CFP and CFP-beta1a-YFP were diffusely distributed in the cytoplasm and highly mobile as indicated by fluorescence recovery after photobleaching. Thus, beta1a does not appear to bind to other cellular proteins in the absence of alpha1S. FRET efficiencies for these cytoplasmic beta1a subunits were approximately 6-7%, consistent with the idea that <10 nm separates the N and C termini. After coexpression with unlabeled alpha1S (in dysgenic or beta1-null myotubes), both constructs produced discrete fluorescent puncta, which correspond to assembled DHPRs in junctions and that did not recover after photobleaching. In beta1-null myotubes, FRET efficiencies of doubly labeled beta1a in puncta were similar to those of the same constructs diffusely distributed in the cytoplasm and appeared to arise intramolecularly, since no FRET was measured when mixtures of singly labeled beta1a (CFP or YFP at the N or C terminus) were expressed in beta1-null myotubes. Thus, DHPRs in tetrads may be arranged such that the N and C termini of adjacent beta1a subunits are located >10 nm from one another. PMID- 16317009 TI - Inhibition of mRNA deadenylation by the nuclear cap binding complex (CBC). AB - Poly(A)-specific ribonuclease (PARN) is a cap-interacting and poly(A)-specific 3' exoribonuclease. Here we have investigated how the cap binding complex (CBC) affects human PARN activity. We showed that CBC, via its 80-kDa subunit (CBP80), inhibited PARN, suggesting that CBC can regulate mRNA deadenylation. The CBC mediated inhibition of PARN was cap-independent, and in keeping with this, the CBP80 subunit alone inhibited PARN. Our data suggested a new function for CBC, identified CBC as a potential regulator of PARN, and emphasized the importance of communication between the two extreme ends of the mRNA as a key strategy to regulate mRNA degradation. Based on our data, we have proposed a model for CBC mediated regulation of PARN, which relies on an interaction between CBP80 and PARN. Association of CBC with PARN might have importance in the regulated recruitment of PARN to the nonsense-mediated decay pathway during the pioneer round of translation. PMID- 16317010 TI - RhoA modulates Smad signaling during transforming growth factor-beta-induced smooth muscle differentiation. AB - We recently reported that transforming growth factor (TGF)-beta induced the neural crest stem cell line Monc-1 to differentiate into a spindle-like contractile smooth muscle cell (SMC) phenotype and that Smad signaling played an important role in this phenomenon. In addition to Smad signaling, other pathways such as mitogen-activated protein kinase (MAPK), phosphoinositol-3 kinase, and RhoA have also been shown to mediate TGF-beta actions. The objectives of this study were to examine whether these signaling pathways contribute to TGF-beta induced SMC development and to test whether Smad signaling cross-talks with other pathway(s) during SMC differentiation induced by TGF-beta. We demonstrate here that RhoA signaling is critical to TGF-beta-induced SMC differentiation. RhoA kinase (ROCK) inhibitor Y27632 significantly blocks the expression of multiple SMC markers such as smooth muscle alpha-actin, SM22alpha, and calponin in TGF beta-treated Monc-1 cells. In addition, Y27632 reversed the cell morphology and abolished the contractility of TGF-beta-treated cells. RhoA signaling was activated as early as 5 min following TGF-beta addition. Dominant negative RhoA blocked nuclear translocation of Smad2 and Smad3 because of the inhibition of phosphorylation of both Smads and inhibited Smad-dependent SBE promoter activity, whereas constitutively active RhoA significantly enhanced SBE promoter activity. Consistent with these results, C3 exotoxin, an inhibitor of RhoA activation, significantly attenuated SBE promoter activity and inhibited Smad nuclear translocation. Taken together, these data point to a new role for RhoA as a modulator of Smad activation while regulating TGF-beta-induced SMC differentiation. PMID- 16317011 TI - A critical role for calponin 2 in vascular development. AB - Calponin 2 (h2 calponin, CNN2) is an actin-binding protein implicated in cytoskeletal organization. We have found that the expression of calponin 2 is relatively restricted to vasculature from 16 to 30 h post-fertilization during zebrafish (Danio rerio) development. Forty-eight hours after injecting antisense morpholino oligos against calponin 2 into embryos at the 1-4-cell stage, zebrafish demonstrated various cardiovascular defects, including sluggish axial and head circulation, absence of circulation in intersegmental vessels and in the dorsal longitudinal anastomotic vessel, enlarged cerebral ventricles, and pericardial edema, in addition to an excess bending, spiraling tail and twisting of the caudal fin. Knockdown of calponin 2 in the Tg(fli1:EGFP)(y1) zebrafish line (in which a fli1 promoter drives vascular-specific enhanced green fluorescent protein expression) indicated that diminished calponin 2 expression blocked the proper migration of endothelial cells during formation of intersegmental vessels. In vitro studies showed that basic fibroblast growth factor-induced human umbilical vein endothelial cell migration was down-regulated by knockdown of calponin 2 expression using an antisense adenovirus, and overexpression of calponin 2 enhanced migration and hastened wound healing. These events were correlated with activation of mitogen-activated protein kinase; moreover, inhibition of this pathway blocked the promigratory effect of calponin 2. Collectively, these data suggest that calponin 2 plays an important role in the migration of endothelial cells both in vivo and in vitro and that its expression is critical for proper vascular development. PMID- 16317012 TI - Dalteparin for prevention of catheter-related complications in cancer patients with central venous catheters: final results of a double-blind, placebo controlled phase III trial. AB - BACKGROUND: Cancer patients receiving chemotherapy experience thromboembolic complications associated with the use of long-term indwelling central venous catheters (CVCs). This prospective, double-blind, placebo-controlled, multicenter study evaluated whether prophylactic treatment with a low molecular weight heparin could prevent clinically relevant catheter-related thrombosis. PATIENTS AND METHODS: Patients with cancer undergoing chemotherapy for at least 12 weeks (n=439) were randomly assigned, in a 2:1 ratio, to receive either dalteparin (5000 IU) or placebo, by subcutaneous injection, once daily for 16 weeks. Patients underwent upper extremity evaluation with either venography or ultrasound at the time of a suspected catheter-related complication (CRC) or upon completion of study medication. The primary end point, as determined by a blinded adjudication committee, was the occurrence of a CRC, defined as the first occurrence of any one of the following: clinically relevant catheter-related thrombosis that was symptomatic or that required anticoagulant or fibrinolytic therapy; catheter-related clinically relevant pulmonary embolism; or catheter obstruction requiring catheter removal. RESULTS: There was no significant difference in the frequency of CRCs between the dalteparin arm (3.7%) and the placebo arm (3.4%; P=0.88), corresponding to a relative risk of 1.0883 (95% confidence interval 0.37-3.19). No difference in the time to CRC was observed between the two arms (P=0.83). There was no significant difference between the dalteparin and placebo groups in terms of major bleeding (1 versus 0) or overall safety. CONCLUSIONS: Dalteparin prophylaxis did not reduce the frequency of thromboembolic complications after CVC implantation in cancer patients. Dalteparin was demonstrated to be safe over 16 weeks of treatment in these patients. PMID- 16317013 TI - Cost-opportunity analysis in clinical oncology: from the 'wild far-west' to a correct integration of the disciplines, avoiding the 'war of the worlds'. PMID- 16317014 TI - A randomised trial in malignant mesothelioma (M) of early (E) versus delayed (D) chemotherapy in symptomatically stable patients: the MED trial. AB - BACKGROUND: Prior phase II trials have demonstrated the therapeutic activity of cytotoxic chemotherapy in mesothelioma. Currently there are few randomised data assessing the role of chemotherapy versus best supportive care (BSC) in the management of patients with stable symptoms after control of any pleural effusion. A policy of observation is often adopted over initial use of chemotherapy. In this prospective randomised trial we assess the use of early versus delayed cytotoxic therapy. The study opened in 1998, and closed in view of a competing national study (MSO 1) in 2003. METHODS: Eligible patients had a performance status3 months and had stable symptoms for at least 4 weeks prior to randomisation. Patients were randomised to receive immediate chemotherapy or initial BSC with the addition of chemotherapy at time of symptomatic progression. All patients received the same platinum-based chemotherapy regimen, MVP [mitomycin C 8 mg/m2 cycles 1, 2, 4 and 6, vinblastine 6 mg/m2, maximum 10 mg, and cisplatin 50 mg/m2 (or carboplatin AUC 5)], every 3 weeks for up to six cycles. RESULTS: A total of 43 patients were recruited, of which 21 were randomised to the early treatment group and 22 to the delayed treatment group. The median ages were 59 years (range 50-78) and 67 years (range 48-75), respectively (P=0.1); other baseline parameters were well matched between the two groups. All 21 patients in the early group received chemotherapy versus 17 patients in the delayed group. Median time to symptomatic progression was 25 weeks in the early group compared with 11 weeks for the delayed group (P=0.1). Median survival was 14 months (1-year survival 66%) for the early group compared with 10 months (1-year survival 36%) for the delayed group (P=0.1). Quality of life was in general better maintained for early treatment and the health resources use was similar in both arms. CONCLUSIONS: In this patient group, presenting with stable symptoms after control of pleural effusion, the early use of chemotherapy provided an extended period of symptom control, and in this small trial a trend to survival advantage. PMID- 16317015 TI - Health and climate change: a call for action. PMID- 16317016 TI - New international consensus on cardiopulmonary resuscitation. PMID- 16317017 TI - Auditory deficits in rats exposed to an environmental PCB mixture during development. AB - Previous studies have indicated that developmental exposure to polychlorinated biphenyls (PCBs) may result in hearing impairment in rats. The cochlea is the suggested site of action, based upon one study demonstrating a loss of outer hair cells on the basilar membrane, and another demonstrating deficits in distortion product otoacoustic emissions (DPOAEs). The current study was conducted to assess the possible ototoxic effects of a unique PCB mixture formulated to model the congener profile of PCBs found in fish consumed by a human population in northeastern Wisconsin. Female Long-Evans rats were dosed orally with the PCB mixture beginning 28 days prior to breeding and continuing until the pups were weaned. Dams were fed one-half of a cookie onto which was pipetted 0, 1, 3, or 6 mg/kg of the PCB mixture dissolved in a corn oil vehicle. On postnatal day (PND) 21, pups were weaned, and one male and one female from each litter were randomly selected for auditory assessment. DPOAEs were measured to assess cochlear function, and auditory brainstem responses (ABRs) were measured to determine effects on central nervous system auditory pathways. DPOAE amplitudes were decreased, and DPOAE and ABR thresholds were elevated across a range of frequencies in PCB-exposed rats. These results support and extend previous reports of auditory impairment in PCB-exposed rats. Developmental exposure to PCBs may also result in subtle auditory impairments in humans, and if so, this may contribute to some of the cognitive deficits that have been observed in epidemiological studies. PMID- 16317018 TI - Patients with hemianopic alexia adopt an inefficient eye movement strategy when reading text. AB - Patients with an acquired homonymous hemianopia often adapt over a period of a few months to compensate for some of the impairments caused by their visual field defect. Changes in their eye movement patterns have been demonstrated as performance on visual tasks improves with time; however, these patients often complain of persistent text reading problems. Using a video-based eye-movement tracking system, we investigated the text reading behaviour of patients with established hemianopic alexia (>6 months post deficit), a condition affecting left-to-right readers, with a homonymous field defect that encroaches into their right foveal/parafoveal visual field. Word-based analyses of text reading are standard in experiments involving normal readers, but this is the first time these methods have been extended to patients with hemianopic alexia. Using this method, we compared the patients' reading scanpaths to those generated by normal controls reading the same passages, and a random model generated by matching the patients' eye movement data to random permutations of the text they read. We demonstrate that patients adopt an inefficient reading strategy, fixating to the left of the preferred viewing location of words of four letters and longer. Fixating to the left of the normal preferred viewing location not only results in less of the fixated word being processed by the language system; ensuing fixations are also more likely to land within the same word (a refixation). It is this refixation rate that is the main factor in slowing reading times in these patients. Our data suggests that patients are able to extract some useful visual information from text to aid the planning of reading scanpaths as their behaviour differs critically from the random model. Potential reasons for this patient group failing to produce an effective reading strategy are discussed. PMID- 16317019 TI - An immune control model for viral replication in the CNS during presymptomatic HIV infection. AB - The brain is targeted by human immunodeficiency virus type 1 (HIV-1) during the course of untreated infection, leading to cognitive impairment, neurological damage and HIV encephalitis (HIVE). To study early dynamics of HIV entry into the brain, we examined a unique autopsy series of samples obtained from 15 untreated individuals who died in the presymptomatic stages of infection from non-HIV causes. HIV was detected and quantified by limiting dilution PCR and genetically characterized in the V3 region of env. Limiting dilution was shown to be essential for correct estimation of genetic partitioning between brain- and lymphoid-associated HIV populations. While no actively expressing HIV-infected cells were detected by immunohistochemistry, variable and generally extremely low levels of proviral DNA were detected in presymptomatic brain samples. V3 region sequences were frequently genetically distinct from lymphoid-associated HIV variants, with association index (AI) values similar to those observed in cases of HIVE. Infiltration of CD8 lymphocytes in the brain was strongly associated with expression of activation markers (MHCII; R = 0.619; P < 0.05), the presence of HIV-infected cells (proviral load; R = 0.608; P < 0.05) and genetic segregation of brain variants from populations in lymphoid tissue (AI value, R = 0.528; P approximately 0.05). CD8 lymphocytes may thus limit replication of HIV seeded into the brain in early stages of infection. Neurological complications in AIDS occur when this control breaks down, due to systemic immunosuppression from HIV that destroys CD8 lymphocyte function and/or through the evolution of more aggressive neuropathogenic variants. PMID- 16317020 TI - Charcot-Marie-Tooth disease type 1A duplication: spectrum of clinical and magnetic resonance imaging features in leg and foot muscles. AB - MRI is an ideal method for identifying areas of muscle atrophy and fatty infiltration. Studies comparing clinical and MRI features of foot and leg muscle atrophy in Charcot-Marie-Tooth disease type 1A (CMT-1A) duplication are lacking. The aim of this study is to describe clinical and MRI patterns of lower limb amyotrophy in CMT-1A. A total of 10 secondary CMT-1A patients and 1 proband patient with de novo mutation were prospectively evaluated. Ages of patients ranged from 8 to 61 years (median, 24). Disease severity in terms of ability to walk and run was established using a nine-point functional disability scale (FDS). We administered the CMT neuropathy score (CMTNS), based on patient's symptoms, neurological examination and neurophysiological testing. Muscle strength of flexo-extensor ankle and toe muscles was assessed manually with the standard Medical Research Council scale. In all 11 patients, leg MRI study included T1- and T2-weighted spin-echo sequences in coronal and axial planes, and a T1-weighted spin-echo sequence with chemical sift fat suppression before and after paramagnetic contrast agent injection. In seven patients both feet were simultaneously studied in coronal and axial planes. Six patients had pes cavus, an FDS score of 0 (normal), mild CMTNS and normal muscle power of foot flexo extensors. In these six patients, MRI showed muscle fatty infiltration of intrinsic foot muscles mainly involving the lumbricals, all four leg muscle compartments being preserved. The remaining five patients had FDS scores from 1 (cramps or fatigability) to 3 (walking difficulty), mild to moderate CMTNS and variable weakness of peroneal musculature. In these five patients MRI showed, besides intrinsic foot muscle involvement, variable and distally accentuated fatty infiltration of the lateral, anterior and superficial posterior leg muscle compartments and, to a lesser degree, of the deep posterior compartment. In four patients muscle oedema and post-contrast enhancement was noted. MRI demonstrated fatty infiltration of clinically normal muscles. We conclude that clinical-MRI patterns of lower limb muscle atrophy vary with evolution of semiology. Selective involvement of intrinsic foot muscles is the characteristic pattern of CMT-1A cases with minimal disease signs. Afterwards this pattern usually combines variable involvement of leg muscles. Our findings help to clarity the pathogenesis of pes cavus in the disease. PMID- 16317021 TI - Goal-driven selective attention in patients with right hemisphere lesions: how intact is the ipsilesional field? AB - Patients with right hemisphere (RH) lesions often display a spatial bias in attention towards the ipsilesional hemifield. The behavioural manifestations of this spatial bias are typically interpreted as reflecting increased or enhanced attention for stimuli within the 'intact' ipsilesional field, and impaired attentional functioning within the contralesional field. In the healthy brain, goal-driven and stimulus-driven attentional processes interact to determine which stimuli should be prioritized for selection. Although unilateral brain damage increases the relative attentional salience of stimuli within the ipsilesional field, it might also cause problems in filtering or attenuating task-irrelevant information. We examined whether goal-driven attention modulates the processing of ipsilesional and contralesional information in 6 patients with unilateral brain damage following RH stroke (5 male, 1 female; mean age 60.8 years) and a group of age and sex-matched controls. We used a flanker task in which participants made speeded judgements on a central target item (a coloured letter). On each trial the target was flanked by a coloured letter in the left and right hemifields. In separate blocks, participants were instructed to judge either the identity or the colour of the central target and to ignore the flankers. The flanker on one side could be congruent, incongruent or neutral with respect to the target, on either the letter or the colour dimension, whereas the flanker on the other side was always neutral on both dimensions. Healthy controls showed significant interference from incongruent flankers on either side. Crucially, however, this effect only occurred for the task-relevant dimension [F(2,10) = 24.60; P < 0.001]. For patients, however, both the task-relevant and task-irrelevant dimensions of ipsilesional flankers interfered with response times [task-relevant: F(2,10) = 7.50, P < 0.05; task-irrelevant: F(1,5) = 6.20, P < 0.05]. Conversely, contralesional flankers influenced response times only when the target and distractor were incongruent on the task-relevant dimension [F(2,10) = 4.85; P < 0.05]. Our findings demonstrate that following RH damage, goal-driven biases cannot constrain the processing of task-irrelevant features of ipsilesional stimuli. We speculate that a lateralized bias in spatial attention leads to unselective prioritization of all feature-based attributes of stimuli appearing within the ipsilesional hemifield, whether or not they are relevant to performance. Attentional selection for ipsilesional stimuli in disorders such as spatial neglect and extinction may not therefore be entirely normal, as previously assumed. PMID- 16317022 TI - Retinal vessel diameters and cerebral small vessel disease: the Rotterdam Scan Study. AB - The direct visualization of retinal vessels provides a unique opportunity to study cerebral small vessel disease, because these vessels share many features. It was reported that persons with smaller retinal arteriolar-to-venular ratio tended to have more white matter lesions on MRI. It is unclear whether this is due to arteriolar narrowing or venular dilatation. We investigated whether smaller arteriolar or larger venular diameters or both were related to severity and progression of cerebral small vessel disease. We studied 490 persons (60-90 years) without dementia from a population-based cohort study. At baseline (1990 1993), retinal arteriolar and venular diameters were measured on digitized images of one eye of each participant. In 1995-1996, participants underwent cerebral MRI scanning. We rated the severity of periventricular white matter lesions on a 9 point scale, approximated a total subcortical white matter lesion volume (range: 0-29.5 ml) and rated the presence of lacunar infarcts. On average 3.3 years later, 279 persons had a second MRI. Changes in periventricular and subcortical white matter lesions were rated with a semi-quantitative scale, and progression was classified as no, minor and marked. An incident infarct was a new infarct on the follow-up MRI. Neither venular nor arteriolar diameters were related to the severity of cerebral small vessel disease. Larger venular diameters were, however, associated with a marked progression of cerebral small vessel disease. Age and gender adjusted odds ratios (ORs) per standard deviation increase were 1.71 [95% confidence interval (CI): 1.11-2.61] for periventricular, 1.72 (95% CI: 1.09-2.71) for subcortical white matter lesion progression and 1.59 (95% CI: 1.06 2.39) for incident lacunar infarcts. These associations were independent of other cardiovascular risk factors. Only the OR for incident lacunar infarcts was attenuated (1.24; 95% CI: 0.72-2.12). No association was observed between arteriolar diameters and progression of cerebral small vessel disease. In conclusion, retinal venular dilatation was related to progression of cerebral small vessel disease. The mechanisms underlying venular dilatation deserve more attention, as they may provide new clues into the pathophysiology of cerebral small vessel disease. PMID- 16317023 TI - Epileptogenicity of cortical dysplasia in temporal lobe dual pathology: an electrophysiological study with invasive recordings. AB - Hippocampal sclerosis is often associated with macroscopic or microscopic dysplasia in the temporal neocortex (TN). The relevance of such a dual pathology with regard to epileptogenesis is unclear. This study investigates the role of both pathologies in the generation of ictal and interictal activity. Ictal (113 seizures) and interictal data from invasive EEG recordings with simultaneous depth electrodes in the hippocampus and subdural electrodes over the TN were analysed retrospectively in 12 patients with variable degrees of hippocampal sclerosis and different types of histologically confirmed temporal cortical dysplasia [all male, age at epilepsy onset <1-29 years (mean 9.6 years), age when invasive recordings were performed 6-50 years (mean 28.2 years)]. Of the seizures 41.3% arose from the amygdala/hippocampus complex (AHC), 34.7% from the TN, 22% were simultaneously recorded from AHC and TN (indeterminate seizure onset), and 2% from other regions. In three patients, seizure onset was recorded only from the AHC. In patients with severe hippocampal sclerosis only 12% of the seizures arose from the TN, whereas in patients with mild hippocampal sclerosis 58% arose from the TN. The type of cortical dysplasia, however, did not predict seizure onset in the AHC or TN. Propagation time from the TN to the AHC tended to be shorter (mean 7.4 s) than vice versa (mean 13.7 s). The most common initial ictal patterns in the AHC were rhythmic beta activity (<25 Hz) and repetitive sharp waves, and in the TN were fast activity (>25 Hz) and repetitive sharp waves. The interictal patterns over the TN were similar to those seen over extratemporal focal cortical dysplasias. Simultaneous recordings from the hippocampus and the TN strongly suggest that dysplastic tissue in the TN is often epileptogenic. The quantitative contribution of the hippocampus to seizure generation corresponded with the degree of hippocampal pathology, whereas different subtypes of cortical dysplasia did not affect its relative contribution to seizure generation and even mild forms of dysplasia were epileptogenic. PMID- 16317024 TI - Cerebellar damage produces selective deficits in verbal working memory. AB - The cerebellum is often active in imaging studies of verbal working memory, consistent with a putative role in articulatory rehearsal. While patients with cerebellar damage occasionally exhibit a mild impairment on standard neuropsychological tests of working memory, these tests are not diagnostic for exploring these processes in detail. The current study was designed to determine whether damage to the cerebellum is associated with impairments on a range of verbal working memory tasks, and if so, under what circumstances. Moreover, we assessed the hypothesis that these impairments are related to impaired rehearsal mechanisms. Patients with damage to the cerebellum (n = 15) exhibited a selective deficit in verbal working memory: spatial forward and backward spans were normal, but forward and backward verbal spans were lower than controls. While the differences were significant, digit spans were relatively preserved, especially in comparison to the dramatic reductions typically observed in classic 'short term memory' patients with perisylvian brain damage. The patients tended to be more impaired on a verbal version compared to a spatial version of a working memory task with a long delay and this impairment was correlated with overall symptom and dysarthria severity. These results are consistent with a contribution of the cerebellum to rehearsal and suggest that inclusion of a delay before recall is especially detrimental in individuals with cerebellar damage. However, when we examined markers of rehearsal (i.e. word-length and articulatory suppression effects) in an immediate serial recall task, we found that qualitative aspects of the patients' rehearsal strategies were unaffected. We propose that the cerebellum may contribute to verbal working memory during the initial phonological encoding and/or by strengthening memory traces rather than by fundamentally subserving covert articulatory rehearsal. PMID- 16317025 TI - The 'essentials' of essential palatal tremor: a reappraisal of the nosology. AB - Essential palatal tremor (EPT) is an uncommon disorder, distinct from symptomatic palatal tremor (SPT), but characterized by superficially similar rhythmic movements of the soft palate. While the pathophysiology of SPT has been relatively well defined, this is not the case in EPT. Based on an analysis of 103 published cases, we reviewed EPT in the context of other movement disorders with similar features and outline possible pathophysiological mechanisms. Phenomenologically it remains best classified as a tremor. Four major causes, including a central generator, peripheral/mechanical, voluntary/special skill and psychogenic, appear to account for the majority of cases of EPT, although there is considerable overlap in the pathogenic mechanisms underlying these categories. Among the cases reviewed, a large proportion fit into the latter two categories, although there are others where multiple mechanisms are likely at play. Based on our reappraisal, we suggest a change in designation to 'isolated palatal tremor', with primary and secondary subtypes. This retains the distinction from SPT and emphasizes the non-uniform, heterogeneous nature of the disorder. PMID- 16317026 TI - Molecular and cellular mechanisms of pharmacoresistance in epilepsy. AB - Epilepsy is a common and devastating neurological disorder. In many patients with epilepsy, seizures are well-controlled with currently available anti-epileptic drugs (AEDs), but a substantial (approximately 30%) proportion of patients continue to have seizures despite carefully optimized drug treatment. Two concepts have been put forward to explain the development of pharmacoresistance. The transporter hypothesis contends that the expression or function of multidrug transporters in the brain is augmented, leading to impaired access of AEDs to CNS targets. The target hypothesis holds that epilepsy-related changes in the properties of the drug targets themselves may result in reduced drug sensitivity. Recent studies have started to dissect the molecular underpinnings of both transporter- and target-mediated mechanisms of pharmacoresistance in human and experimental epilepsy. An emerging understanding of these underlying molecular and cellular mechanisms is likely to provide important impetus for the development of new pharmacological treatment strategies. PMID- 16317027 TI - Knee ligament mechanical properties are not influenced by estrogen or its receptors. AB - Women are at greater risk of tearing their knee anterior cruciate ligament (ACL) than men participating in similar athletic activities. There is currently no conclusive explanation for this disparity; however, as ACL injuries in women have been linked with estrogen fluctuations during the menstrual cycle, one hypothesis is that estrogen has a direct detrimental effect on knee ligament mechanical properties. This study investigated the influence of estrogen and its receptors (ER alpha and ER beta) on knee ligament mechanical properties. This was achieved by testing the viscoelastic and tensile mechanical properties of knee medial collateral ligaments (MCL) and ACLs from: 1) male Sprague-Dawley rats treated with either estrogen (17alpha-ethynylestradiol; 0.03 mg/kg) or an ER alpha specific agonist (propyl pyrazole triol; 2 mg/kg), and 2) female mice with a null mutation of the gene encoding for ER beta. Estrogen treatment had no significant effects on the viscoelastic or tensile mechanical properties of the rat MCL or ACL. Similarly, pharmacological stimulation of ER alpha using a selective agonist in rats and genetic modulation of ER beta by null mutation of its gene in mice did not influence MCL or ACL properties. These data indicate that estrogen does not have a major direct effect on ligament mechanical properties. Energies for the prevention of the disproportionately high rate of knee ligament injuries in women may be better spent focusing on more established and modifiable risk factors, such as abnormalities in neuromuscular control about the knee. PMID- 16317028 TI - Amelioration of oxidant stress by the defensin lysozyme. AB - Reactive oxidant species (ROS), products of normal metabolism, cause oxidant injury if they accumulate in pathological amounts. Lysozyme (LZ) contains an 18 amino acid domain that binds agents such as advanced glycation end products (AGE) that generate ROS. We examined whether endogenous LZ affected physiological, or baseline, antioxidant balance and provided protection against both acute and chronic oxidant injury, using paraquat and H2O2 as agents of acute injury and AGE for chronic injury. Hen egg LZ-Tg mice had threefold higher serum LZ levels and decreased baseline AGE levels in serum and liver. These findings were linked to an enhanced baseline systemic GSH-to-GSSG ratio. Baseline levels of stress response genes p66(Shc) and c-Jun were also lower in liver tissue of LZ-Tg mice. Survival from severe oxidant injury induced by paraquat was twofold greater in LZ Tg mice. In addition, LZ-Tg mice were resistant to chronic exogenous oxidant stress (OS) induced by AGE administration. Preincubation of hepatocytes (Hep G2) with LZ suppressed redox balance at baseline, as well as OS after added paraquat, AGE, or H2O2. LZ also ameliorated paraquat-enhanced cell apoptosis in a dose dependent manner and suppressed AGE-induced p66(Shc) expression and c-Jun phosphorylation in Hep G2 cells. Thus LZ provides protection against acute and chronic oxidant injury by mechanisms involving suppression of ROS generation and of OS response genes. PMID- 16317029 TI - Mu and delta, but not kappa, opioid agonists induce spastic paraparesis after a short period of spinal cord ischaemia in rats. AB - BACKGROUND: Intrathecal (IT) morphine given after a short interval of aortic occlusion in a rodent model induced transient spastic paraparesis via opioid receptor-predicted actions in spinal cord. To determine the role(s) of spinal opioid receptor subtypes we investigated whether IT administration of various selective opioid receptor agonists can induce paraparesis following a short period of spinal cord ischaemia in rats. METHODS: In Sprague-Dawley rats implanted with an IT catheter, spinal cord ischaemia was induced for 6 min using an intraaortic balloon. Mu ([D-Ala2, N-Me Phe4, Gly-ol5] enkephalin), kappa (U50488H) or delta ([D-Pen(2,5)] enkephalin) selective agonists were injected intrathecally 30 min after reperfusion. A separate group of animals was used to investigate the dose-response effect on this motor dysfunction. For this purpose, three doses of mu, kappa, or delta agonists were injected intrathecally after ischaemia. After IT injection, recovery of motor function was assessed periodically using the motor deficit index (0=complete recovery; 6=complete paraplegia). RESULTS: IT administration of mu and delta but not kappa agonists produced dose-dependent effects in the induction of spastic paraparesis. In addition, this spasticity induced by IT mu and delta agonists was reversed completely by IT naloxone and naltrindole, respectively. CONCLUSION: These results suggest that the effect of various opioids on motor function after a short period of spinal cord ischaemia depends upon individual opioid receptor subtypes. PMID- 16317030 TI - The suppression of spinal F-waves by propofol does not predict immobility to painful stimuli in humans. AB - BACKGROUND: The immobilizing effects of volatile anaesthetics are primarily mediated at the spinal level. A suppression of recurrent spinal responses (F waves), which reflect spinal excitability, has been shown for propofol. We have assessed the concentration-dependent F-wave suppression by propofol and related it to the logistic regression curve for suppression of movement to noxious stimuli and the effect on the bispectral index (BIS). The predictive power of drug effects on F-waves and BIS for movement responses to noxious stimuli was tested. METHODS: In 24 patients anaesthesia was induced and maintained with propofol infused by a target controlled infusion pump at stepwise increasing and decreasing plasma concentrations between 0.5 and 4.5 mg litre(-1). The F-waves of the abductor hallucis muscle were recorded at a frequency of 0.2 Hz. BIS values were recorded continuously. Calculated propofol concentrations and F-wave amplitude and persistence were analyzed in terms of a pharmacokinetic pharmacodynamic (PK/PD) model with a simple sigmoid concentration-response function. Motor responses to tetanic electrical stimulation (50 Hz, 60 mA, 5 s, volar forearm) were tested and the EC(50tetanus) was calculated using logistic regression. RESULTS: For slowly increasing propofol concentrations, computer fits of the PK/PD model for the suppression by propofol yielded a median EC50 of 1.26 (0.4-2.3) and 1.9 (1.0-2.8) mg litre(-1) for the F-wave amplitude and persistence, respectively. These values are far lower than the calculated EC(50) for noxious electrical stimulation of 3.75 mg litre(-1). This difference results in a poor prediction probability of movement to noxious stimuli of 0.59 for the F wave amplitude. CONCLUSIONS: F-waves are almost completely suppressed at subclinical propofol concentrations and they are therefore not suitable for prediction of motor responses to noxious stimuli under propofol mono-anaesthesia. PMID- 16317031 TI - Fluid absorption in endoscopic surgery. AB - Fluid absorption is an unpredictable complication of endoscopic surgery. Absorption of small amounts of fluid (1-2 litre) occurs in 5-10% of patients undergoing transurethral prostatic resection and results in an easily overlooked mild transurethral resection (TUR) syndrome. Large-scale fluid absorption is rare but leads to symptoms severe enough to require intensive care. Pathophysiological mechanisms consist of pharmacological effects of the irrigant solutes, the volume effect of the irrigant water, dilutional hyponatraemia and brain oedema. Other less widely known factors include absolute losses of sodium by urinary excretion and morphological changes in the heart muscle, both of which promote a hypokinetic circulation. Studies in animals, volunteers and patients show that irrigation with glycine solution should be avoided. Preventive measures, such as low-pressure irrigation, might reduce the extent of fluid absorption but does not eliminate this complication. Monitoring the extent of absorption during surgery allows control of the fluid balance in the individual patient, but such monitoring is not used widely. However, the anaesthetist must be aware of the symptoms and be able to diagnose this complication. Treatment should be based on administration of hypertonic saline rather than on diuretics. New techniques, such as bipolar resectoscopes and vaporizing instead of resecting tissue, result in a continuous change of the prerequisites for fluid absorption and its consequences. PMID- 16317032 TI - Preventing malaria in pregnancy: a study of perceptions and policy implications in Mukono district, Uganda. AB - Although the efficacy of insecticide-treated nets (ITNs) in malaria prevention is well documented, the low coverage of ITNs in malaria endemic countries necessitates investigation on factors that limit access to this intervention. An exploratory study was conducted in Mukono district, Uganda, to assess perceptions and use of ITNs. Results show that malaria is perceived as a serious illness among pregnant women and children, and there is high awareness on the benefits of ITNs. However, ITNs are used by few people, mainly because of their high cost and the perception that the chemicals used to treat them have dangerous effects on pregnancy and the foetus. Other factors that influence the use of ITNs include low utilization of antenatal care, husband's lack of interest in malaria prevention and the perception that adolescent girls and primigravidae are at a low risk of getting malaria. The policy implications of these findings include demystifying the negative perceptions on the chemicals used to treat nets and subsidizing the cost of ITNs in order to increase access to them. These findings provide important lessons for malaria control programmes that aim at increasing access to ITNs by pregnant women in developing countries. PMID- 16317033 TI - Positional information and mobile transcriptional regulators determine cell pattern in the Arabidopsis root epidermis. AB - The root epidermis is a model system for deciphering the mechanism underpinning the formation of cellular pattern in planar groups of cells. The epidermis comprises rows of hair cells (H) and non-hair-bearing epidermal cells (N). Laser ablation and clonal analysis have shown that the fates of epidermal cells are flexible through development and that positional information which may be located in the cell wall or extracellular matrix determines cell fate. A leucine-rich repeat protein called SCRAMBLED is required for the development of cell pattern which may be involved in the perception of positional information. It is proposed that positional signals then initiate the cell-specific expression of a number of transcription factors that complete the patterning process, resulting in the expression of hair-promoting genes in hair cells (H) and their repression in the hairless cells (N). PMID- 16317034 TI - Identification of a mitochondrial ATP synthase small subunit gene (RMtATP6) expressed in response to salts and osmotic stresses in rice (Oryza sativa L.). AB - Large areas of northern China have alkaline soil due to the accumulation of sodium carbonates (NaHCO3, Na2CO3). To understand better how plants can tolerate alkaline soil, a cDNA library was prepared from rice (Oryza sativa L.) roots grown in the presence of NaHCO3 stress. A cDNA clone isolated from this library was identified by a homology search as a mitochondrial ATP synthase 6 kDa subunit gene (RMtATP6; GenBank accession nos AB055076, BAB21526). In transformed yeast and tobacco protoplasts, the RMtATP6 protein was localized in mitochondria using the green fluorescent protein (GFP) marker. Analysis of RMtATP6 mRNA levels suggested that the expression of this gene was induced by stress from sodium carbonates and other sodium salts. Transgenic tobacco overexpressing the RMtATP6 gene had greater tolerance to salt stress at the seedling stage than untransformed tobacco. Among the other genes for F1F0-ATPase of rice, some were found to be up-regulated by some environmental stresses and some were not. These data suggest that the RMtATP6 protein acts as a subunit of ATP synthase, and is expressed in response to stress from several salts, with the other genes coding for the subunits of the same ATP-synthase. PMID- 16317035 TI - The novel cytochrome c6 of chloroplasts: a case of evolutionary bricolage? AB - Cytochrome c6 has long been known as a redox carrier of the thylakoid lumen of cyanobacteria and some eukaryotic algae that can substitute for plastocyanin in electron transfer. Until recently, it was widely accepted that land plants lack a cytochrome c6. However, a homologue of the protein has now been identified in several plant species together with an additional isoform in the green alga Chlamydomonas reinhardtii. This form of the protein, designated cytochrome c6A, differs from the 'conventional' cytochrome c6 in possessing a conserved insertion of 12 amino acids that includes two absolutely conserved cysteine residues. There are conflicting reports of whether cytochrome c6A can substitute for plastocyanin in photosynthetic electron transfer. The evidence for and against this is reviewed and the likely evolutionary history of cytochrome c6A is discussed. It is suggested that it has been converted from a primary role in electron transfer to one in regulation within the chloroplast, and is an example of evolutionary 'bricolage'. PMID- 16317036 TI - Dual targeting of xylanase to chloroplasts and peroxisomes as a means to increase protein accumulation in plant cells. AB - One of the limiting factors in the production of recombinant proteins in transgenic plants is the low level of protein accumulation. A strategy was investigated for a high level of protein accumulation in plant cells. A fungal xylanase encoded by XYLII of Trichoderma reesei was chosen as the model protein because xylanases have a high potential for applications in environment-related technologies. Xylanase was expressed in the cytosol or targeted either to chloroplasts or peroxisomes alone, or to both organelles simultaneously. When xylanase was targeted to both chloroplasts and peroxisomes simultaneously the amount of xylanase accumulated was 160% of that in chloroplasts alone and 240% of that in peroxisomes alone although the transcript levels were similar among these constructs. The growth stage of the transgenic plants also affected the total amount of xylanase; the highest level of accumulation occurred at the time of flowering. This study provides genetic and biochemical data demonstrating that a high level of protein accumulation in transgenic plants can be obtained by targeting a protein to both chloroplasts and peroxisomes at the same time. PMID- 16317037 TI - High apoplastic solute concentrations in leaves alter water relations of the halophytic shrub, Sarcobatus vermiculatus. AB - Predawn plant water potential (Psi(w)) is used to estimate soil moisture available to plants because plants are expected to equilibrate with the root-zone Psi(w). Although this equilibrium assumption provides the basis for interpreting many physiological and ecological parameters, much work suggests predawn plant Psi(w) is often more negative than root-zone soil Psi(w). For many halophytes even when soils are well-watered and night-time shoot and root water loss eliminated, predawn disequilibrium (PDD) between leaf and soil Psi(w) can exceed 0.5 MPa. A model halophyte, Sarcobatus vermiculatus, was used to test the predictions that low predawn solute potential (Psi(s)) in the leaf apoplast is a major mechanism driving PDD and that low Psi(s) is due to high Na+ and K+ concentrations in the leaf apoplast. Measurements of leaf cell turgor (Psi(p)) and solute potential (Psi(s)) of plants grown under a range of soil salinities demonstrated that predawn symplast Psi(w) was 1.7 to 2.1 MPa more negative than predawn xylem Psi(w), indicating a significant negative apoplastic Psi(s). Measurements on isolated apoplastic fluid indicated that Na+ concentrations in the leaf apoplast ranged from 80 to 230 mM, depending on salinity, while apoplastic K+ remained around 50 mM. The water relations measurements suggest that without a low apoplastic Psi(s), predawn Psi(p) may reach pressures that could cause cell damage. It is proposed that low predawn apoplastic Psi(s) may be an efficient way to regulate Psi(p) in plants that accumulate high concentrations of osmotica or when plants are subject to fluctuating patterns of soil water availability. PMID- 16317038 TI - Update on ureide degradation in legumes. AB - Warm season N2-fixing legumes move fixed N from the nodules to the aerial portions of the plant primarily in the form of ureides, allantoin and allantoate, oxidation products of purines synthesized de novo in the nodule. Ureides are also products of purine turnover in senescing tissues, such as seedling cotyledons. A combination of biochemical and molecular approaches in both crop and model species has shed new light on the metabolic pathways involved in both the synthesis and degradation of allantoin. Improved understanding of ureide biochemistry includes two 'additional' enzymatic steps in the conversion of uric acid to allantoin in the nodule and the mechanism of allantoin and allantoate breakdown in leaf tissue. Ureide accumulation and metabolism in leaves have also been implicated in the feedback inhibition of N2-fixation under water limitation. Sensitivity to water deficit differs among soybean cultivars. Manganese supplementation has been shown to modify relative susceptibility or tolerance to this process in a cultivar-dependent manner. A discussion of the potential roles for ureides and manganese in the feedback inhibition of N2-fixation under water limitation is presented. The existing data are examined in relation to potential changes in both aerial carbon and nitrogen supply under water deficit. PMID- 16317039 TI - Expression of plant cyclic nucleotide-gated cation channels in yeast. AB - The functional properties of inwardly conducting plant cyclic nucleotide-gated cation channels (CNGCs) have not been thoroughly characterized due in part to the recalcitrance of their functional expression in heterologous systems. Here, K+ uptake-deficient mutants of yeast (trk1,2) and Escherichia coli (LB650), as well as the Ca2+-uptake yeast mutant mid1,cch1, were used for functional characterization of Arabidopsis thaliana CNGCs, with the aim of identifying some of the cultural and physiological conditions that impact on plant CNGC function in heterologous systems. Use of the Ca2+-uptake yeast mutant provided the first evidence consistent with Ca2+ conduction by the A. thaliana CNGC AtCNGC1. Expression of AtCNGC1 in LB650 demonstrated that mutants of Escherichia coli (which has no endogenous calmodulin) can also be used to study functional properties of CNGCs. Expression of AtCNGC2 and AtCNGC4 enhanced growth of trk1,2 in the presence of hygromycin; AtCNGC1 has less of an effect. Deletion of the AtCNGC1 calmodulin-binding domain enhanced growth of trk1,2 at low external K+ but not of LB650, suggesting that yeast calmodulin may bind to, and down-regulate this plant channel. In vitro binding studies confirmed this physical interaction. Northern analysis, green fluorescent protein:AtCNGC1 fusion protein expression, as well as an antibody raised against a portion of AtCNGC1, were used to monitor expression of AtCNGC1 and deletion constructs of the channel in the heterologous systems. In the presence of the activating ligand cAMP, expression of the AtCNGC1 channel with the calmodulin-binding domain deleted increased intracellular [K+] of trk1,2. Trk1,2 is hypersensitive to the toxic cations spermine, tetramethylamine, and NH4+. These compounds, as well as amiloride, inhibited trk1,2 growth and thereby improved the efficacy of this yeast mutant as a heterologous expression system for CNGCs. In addition to characterizing mutants of yeast and E. coli as assay systems for plant CNGCs, work presented in this report demonstrates, for the first time, that a plant CNGC can retain ion channel function despite (partial) deletion of its calmodulin-binding domain and that yeast calmodulin can bind to and possibly down-regulate a plant CNGC. PMID- 16317040 TI - Exploitation of genomic sequences in a systematic analysis to access how cyanobacteria sense environmental stress. AB - The perception and subsequent transduction of environmental signals are primary events in the acclimation of living organisms to changes in their environment. Many of the molecular sensors and transducers of environmental stress cannot be identified by traditional and conventional methods. Therefore, the genomic information has been exploited in a systematic approach to this problem, performing systematic mutagenesis of potential sensors and transducers, namely, histidine kinases and response regulators, respectively, in combination with DNA microarray analysis, to examine the genome-wide expression of genes in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Using targeted mutagenesis, 44 out of the 47 histidine kinases and 42 out of the 45 response regulators of this organism have successfully been inactivated. The resultant mutant libraries were screened by genome-wide DNA microarray analysis and by slot blot hybridization analysis under various stress and non-stress conditions. Histidine kinases have been identified that perceive and transduce signals of low temperature, hyperosmotic, and salt stress, as well as manganese deficiency. PMID- 16317041 TI - Root responses to soil physical conditions; growth dynamics from field to cell. AB - Root growth in the field is often slowed by a combination of soil physical stresses, including mechanical impedance, water stress, and oxygen deficiency. The stresses operating may vary continually, depending on the location of the root in the soil profile, the prevailing soil water conditions, and the degree to which the soil has been compacted. The dynamics of root growth responses are considered in this paper, together with the cellular responses that underlie them. Certain root responses facilitate elongation in hard soil, for example, increased sloughing of border cells and exudation from the root cap decreases friction; and thickening of the root relieves stress in front of the root apex and decreases buckling. Whole root systems may also grow preferentially in loose versus dense soil, but this response depends on genotype and the spatial arrangement of loose and compact soil with respect to the main root axes. Decreased root elongation is often accompanied by a decrease in both cell flux and axial cell extension, and recent computer-based models are increasing our understanding of these processes. In the case of mechanical impedance, large changes in cell shape occur, giving rise to shorter fatter cells. There is still uncertainty about many aspects of this response, including the changes in cell walls that control axial versus radial extension, and the degree to which the epidermis, cortex, and stele control root elongation. Optical flow techniques enable tracking of root surfaces with time to yield estimates of two-dimensional velocity fields. It is demonstrated that these techniques can be applied successfully to time-lapse sequences of confocal microscope images of living roots, in order to determine velocity fields and strain rates of groups of cells. In combination with new molecular approaches this provides a promising way of investigating and modelling the mechanisms controlling growth perturbations in response to environmental stresses. PMID- 16317042 TI - The co-ordination of cell division, differentiation and morphogenesis in the shoot apical meristem: a perspective. AB - Whether morphogenesis is cell division-driven or organismal-based has been a long running debate in plant biology. This article is a summary of a series of experiments aimed at distinguishing these alternate views by local manipulation of parameters of cell division frequency, orientation, and growth within the shoot apical meristem. These data, put in the context of other investigations in this area, support an organismal view of plant morphogenesis and support the idea that the cell wall plays a key role in the mechanism by which this is achieved. At the same time, the data indicate that the intimate but variable relationship between cell growth and division within the organism means that cell proliferation can indirectly influence this process, leading to a context dependent influence on morphogenesis. Finally, cell growth and proliferation are intimately related with the process of differentiation as cells exit the meristem. In the final part of the article the molecular mechanism by which these basic cellular parameters are intertwined is discussed. PMID- 16317043 TI - Role of the subcellular localization of ALK tyrosine kinase domain in neuronal differentiation of PC12 cells. AB - Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase essentially and transiently expressed in specific areas of the developing central and peripheral nervous systems. We previously demonstrated that a membrane-bound and constitutively active form of the ALK protein tyrosine kinase (PTK) domain induced the neuron-like differentiation of PC12 cells through specific activation of the mitogen-activated protein kinase (MAP kinase) pathway. Its PTK domain had been originally identified in a nucleo-cytosolic and constitutively active transforming protein, NPM-ALK. Downstream targets involved in oncogenic proliferation and survival processes have been proposed to include phospholipase Cgamma (PLCgamma), phosphoinositide 3-kinase (PI 3-kinase)/AKT, STAT 3/5 and Src. We therefore postulated that activation of specific signaling pathways leading to differentiation or proliferation can be differently controlled depending on the subcellular localization of ALK PTK domain. To increase knowledge of its physiological role in the nervous system, we focused in the present study on the influence of its subcellular localization on neuronal differentiation. To achieve this goal, we characterized biological responses and transduction pathways in PC12 cells elicited by various constructs encoding membrane-bound (through transmembrane or myristyl sequences) or cytosolic ALK-derived proteins. In order to control the activation of their PTK domain, we used an inducible dimerization system. Here, we demonstrate that membrane attachment of the ALK PTK domain, in PC12 cells, is crucial for initiation of neurite outgrowth and proliferation arrest through a decrease of DNA synthesis. Furthermore, we show that this differentiation process relies on specific and sustained activation of ERK 1/2 proteins. By contrast, activation of the cytosolic form of this domain fails to induce MAP kinase activation and cell differentiation but promotes a PI 3 kinase/AKT-dependent PC12 cell proliferation. These data indicate that subcellular localization of the ALK PTK domain was a determinant for the control and specificity of downstream transduction cascades and was crucial for deciding the fate to which the neuronal cell will be committed. PMID- 16317044 TI - Paxillin is essential for PTP-PEST-dependent regulation of cell spreading and motility: a role for paxillin kinase linker. AB - The tyrosine phosphatase PTP-PEST has been implicated in the regulation of cell spreading and migration through dephosphorylation of focal adhesion proteins and inhibition of Rac GTPase activity. The focal adhesion adaptor protein paxillin is also necessary for normal cell migration and binds directly to PTP-PEST. In this study, we have utilized PTP-PEST(-/-) and paxillin(-/-) fibroblasts to demonstrate that paxillin is essential for PTP-PEST inhibition of cell spreading and membrane protrusion as well as inhibition of adhesion-induced Rac activation. Furthermore, we show that paxillin-binding is necessary for PTP-PEST stimulation of cell migration. Mutation analysis indicates that PTP-PEST function involves binding to the paxillin C-terminal LIM domains, and signaling through the tyrosine 31 and 118 phosphorylation sites, as well as the LD4 motif of the paxillin N-terminus. Using 'substrate trapping' approaches and immunoprecipitation, we show that the ARF GAP paxillin kinase linker PKL/GIT2, a paxillin LD4 binding partner, is a substrate for PTP-PEST. Additionally, the PKL paxillin interaction was necessary for PTP-PEST inhibition of cell spreading. These data provide mechanistic insight into how the paxillin-PTP-PEST interaction contributes to integrin signaling events associated with the spatiotemporal regulation of key modulators of the cytoskeleton and cell motility machinery. PMID- 16317045 TI - TLS facilitates transport of mRNA encoding an actin-stabilizing protein to dendritic spines. AB - TLS (translocation in liposarcoma), an RNA-binding protein, was originally identified as a heterogeneous ribonuclear protein (hnRNP). Recently, we showed that TLS is localized in neuronal dendrites of mouse hippocampal neurons and is translocated to the spines, where local translation takes place, in an mGluR5 activation-dependent manner. However, the specific role of TLS has not been clarified. TLS-null neurons display abnormal spine morphology, suggesting that TLS-deficiency may impair activity-dependent actin reorganization in spines. To address this issue, we screened for mouse brain transcripts by their in vitro binding to TLS, and identified RNAs that associate with TLS, including mRNAs encoding actin-related proteins such as actin-stabilizing protein Nd1-L. Nd1-L transcripts were increased in the dendrites upon mGluR activation and significantly reduced in TLS-null dendrites. Overexpression of Nd1-L in mouse hippocampal neurons prevented damage to spine structure caused by actin destabilization. Our results demonstrate that TLS associates with mRNA encoding an actin-related protein and may be involved in actin reorganization in spines. PMID- 16317046 TI - Histone acetylation increases chromatin accessibility. AB - In eukaryotes, the interaction of DNA with proteins and supramolecular complexes involved in gene expression is controlled by the dynamic organization of chromatin inasmuch as it defines the DNA accessibility. Here, the nuclear distribution of microinjected fluorescein-labeled dextrans of 42 kDa to 2.5 MDa molecular mass was used to characterize the chromatin accessibility in dependence on histone acetylation. Measurements of the fluorescein-dextran sizes were combined with an image correlation spectroscopy analysis, and three different interphase chromatin condensation states with apparent pore sizes of 16-20 nm, 36 56 nm and 60-100 nm were identified. A reversible change of the chromatin conformation to a uniform 60-100 nm pore size distribution was observed upon increased histone acetylation. This result identifies histone acetylation as a central factor in the dynamic regulation of chromatin accessibility during interphase. In mitotic chromosomes, the chromatin exclusion limit was 10-20 nm and independent of the histone acetylation state. PMID- 16317047 TI - Ace2p contributes to fission yeast septin ring assembly by regulating mid2+ expression. AB - The fission yeast Schizosaccharomyces pombe divides through constriction of an actomyosin-based contractile ring followed by formation and degradation of a medial septum. Formation of an organized septin ring is also important for the completion of S. pombe cell division and this event relies on the production of Mid2p. mid2+ mRNA and protein accumulate in mitosis. Recent microarray analyses identified mid2+ as a target of the Ace2p transcription factor, and ace2+ as a target of the Sep1p transcription factor. In this study, we find that Mid2p production is controlled by Ace2p functioning downstream of Sep1p. Consequently, both Sep1p and Ace2p are required for septin ring assembly and genetic analyses indicate that septin rings function in parallel with other Ace2p targets to achieve efficient cell division. Conversely, forced overproduction of Sep1p or Ace2p prevents septin ring disassembly. We find that Ace2p levels peak during anaphase and Ace2p is post-translationally modified by phosphorylation and ubiquitylation. Ace2p localizes symmetrically to dividing nuclei and functions independently of the septation initiation network. PMID- 16317048 TI - Consequences of loss of PINCH2 expression in mice. AB - PINCH2 belongs, together with PINCH1, to a new family of focal adhesion proteins, the members of which are composed of five LIM domains. PINCH1 and PINCH2 interact, through their first LIM domain, with the integrin-linked kinase and thereby link integrins with several signal transduction pathways. Despite their high similarity, it has been shown that PINCH1 and PINCH2 could exert distinct functions during cell spreading and cell survival. To investigate the function of PINCH2 in vivo, we deleted PINCH2 in mouse using the loxP/Cre system. In contrast to the PINCH1-deficient mice, which die at the peri-implantation stage, PINCH2 null mice are viable, fertile and show no overt phenotype. Histological analysis of tissues that express high levels of PINCH2 such as bladder and kidney revealed no apparent abnormalities, but showed a significant upregulation of PINCH1, suggesting that the two PINCH proteins may have, at least in part, overlapping function in vivo. To further test this possibility, we established PINCH1-null mouse embryonic fibroblasts, which express neither PINCH1 nor PINCH2. We found that in fibroblasts with a PINCH1/2-null background, PINCH2 is able to rescue the spreading and adhesion defects of mutant fibroblasts to the same extent as PINCH1. Furthermore, we show that the LIM1 domain only of either PINCH1 or PINCH2 can prevent ILK degradation despite their failure to localize to focal adhesions. Altogether these results suggest that PINCH1 and PINCH2 share overlapping functions and operate dependently and independently of their subcellular localization. PMID- 16317049 TI - Calcium increases endocytotic vesicle size and accelerates membrane fission in insulin-secreting INS-1 cells. AB - In many cells, endocytotic membrane retrieval is accelerated by Ca2+. The effect of Ca2+ on single endocytotic vesicles and fission pore kinetics was examined by measuring capacitance and conductance changes in small membrane patches of insulin-secreting INS-1 cells. In intact cells, elevation of Ca2+ by glucose stimulation induced a 1.8-fold increase in membrane internalisation. This surprisingly resulted from an increased unitary capacitance of endocytotic vesicles whereas the frequency of endocytosis was unaltered. This effect of glucose was prevented by inhibition of L- or R-type Ca2+ channels. Extracellular (pipette) Ca2+ was found to regulate endocytotic vesicle capacitance in a bimodal manner. Vesicle capacitance was increased at intermediate Ca2+ (2.6 mM), but not at high Ca2+ (10 mM). Similar results were obtained upon direct application of 100 nM and 0.5 mM Ca2+ to the intracellular surface of inside-out excised membrane patches, and in these experiments the increase in vesicle capacitance was prevented by the calcineurin inhibitor deltamethrin. Endocytotic fission pore kinetics were accelerated by Ca2+ in both the intact cells and isolated membrane patches; however, the effect in this case was neither bimodal nor deltamethrin sensitive. Membrane retrieval can therefore be upregulated by a Ca2+-dependent increase in endocytotic vesicle size and acceleration of membrane fission in insulin-secreting INS-1 cells. PMID- 16317050 TI - Regulation of two insulin granule populations within the reserve pool by distinct calcium sources. AB - Insulin granule trafficking is a key step of glucose-stimulated insulin secretion from pancreatic beta cells. Using quantitative live cell imaging, we examined insulin granule movements within the reserve pool upon secretory stimulation in betaTC3 cells. For this study, we developed a custom image analysis program that permitted automatic tracking of the individual motions of over 20,000 granules. This analysis of a large sample size enabled us to study micro-populations of granules that were not quantifiable in previous studies. While over 90% of the granules depend on Ca2+ efflux from the endoplasmic reticulum for their mobilization, a small and fast-moving population of granules responds to extracellular Ca2+ influx after depolarization of the plasma membrane. We show that this differential regulation of the two granule populations is consistent with localized Ca2+ signals, and that the cytoskeletal network is involved in both types of granule movement. The fast-moving granules are correlated temporally and spatially to the replacement of the secreted insulin granules, which supports the hypothesis that these granules are responsible for replenishing the readily releasable pool. Our study provides a model by which glucose and other secretory stimuli can regulate the readily releasable pool through the same mechanisms that regulate insulin secretion. PMID- 16317051 TI - Active ERK1 is dimerized in vivo: bisphosphodimers generate peak kinase activity and monophosphodimers maintain basal ERK1 activity. AB - ERK1 and ERK2 are widely involved in cell signalling. Using a recombinant approach, it has been shown that exogenous ERK2 is capable of dimerization and that preventing dimerization reduces its nuclear accumulation on stimulation. Dimerization occurs on phosphorylation; the dimer partner of phosphorylated ERK2 may be either phosphorylated or unphosphorylated. It has been assumed that monophosphodimers are hemiactive. Here we show that ERK1 is capable of dimerization both in vivo and in vitro. Dimerization of human recombinant ERK1 in vitro requires both ERK1 phosphorylation and cellular cofactor(s); it leads to the formation of a high molecular weight complex that can be dissociated by treatment with beta-mercaptoethanol. We demonstrate for the first time in both sea urchin embryos and human cells that native ERK forms dimers and that high ERK kinase activity is largely associated with bisphosphodimers, not with monophosphodimers or phosphorylated monomers. The activity of the bisphosphodimer is about 20-fold higher than that of the phosphorylated monomer in vitro and the bisphosphodimer shows 5- to 7-fold higher in vivo activity than the basal activity attributable to the monophosphodimer. Thus phosphorylation of both partners in the dimer is a hallmark of ERK activation. Judgments made about ERK kinase activity associated with phosphorylated monomers are at best a proxy for ERK activity. PMID- 16317052 TI - The best of times and the worst of times: A conversation with Vicky Gregg. Interview by James C Robinson. AB - Blue Cross and Blue Shield plans have enjoyed enviable financial success, while much of the health care delivery system has faced severe financial strains. The contrast is striking in Tennessee, where BlueCross BlueShield of Tennessee dominates the commercial health insurance market and administers much of the state's Medicaid program, TennCare, while TennCare itself recently dropped hundreds of thousands of enrollees from coverage because of budgetary constraints. CEO Vicky Gregg draws lessons from the TennCare crisis for other states; describes her organization's investment in information technology, pricing policies, and management of new clinical technologies; and explains the continued political popularity of a profitable nonprofit health plan. PMID- 16317053 TI - Strategies for the characterization of disorders in cortisol sensitivity. AB - CONTEXT: The clinical presentation of abnormalities in glucocorticoid (GC) sensitivity is diverse, and therefore it is difficult to diagnose this condition. OBJECTIVE AND DESIGN: The objective of the study was to develop strategies for the characterization of GC sensitivity disorders. SETTING: The study was conducted in an outpatient clinic. PATIENTS: Nine patients with GC sensitivity disorders participated. INTERVENTIONS: Sequence analysis of the GC receptor (GR), determination of GR number per cell, GR ligand-binding affinity, and GR splice regulation were performed in freshly prepared peripheral blood mononuclear lymphocytes and Epstein-Barr virus-transformed lymphoblasts. Cellular GC sensitivity was determined ex vivo by measuring the effect of dexamethasone on GC induced leucine-zipper and IL-2 mRNA levels and on cell proliferation. RESULTS: Differences in GR number per cell, GR affinity, GR splice variants, and effects on transactivation or transrepression of GC-sensitive genes were observed between patients and controls. Epstein-Barr virus transformation of lymphoblasts had no influence on GR affinity but increased the GR number 5-fold in healthy controls. In patients diagnosed as cortisol resistant, however, GR number after transformation was increased significantly less than 5-fold, whereas a higher GR number was observed in a patient suspected of cortisol hypersensitivity. CONCLUSION: This study illustrates several strategies to define abnormalities in GC sensitivity by describing nine patients with affected GC sensitivity, all with a unique clinical course and background. PMID- 16317054 TI - 123I thyroid uptake and thyroid size at 24, 48, and 72 hours after the administration of recombinant human thyroid-stimulating hormone to normal volunteers. AB - CONTEXT: Recombinant human TSH (rhTSH) is used to evaluate thyroid carcinoma patients and off-label for (131)I thyroid ablation and nontoxic goiter therapy. OBJECTIVE: Our objective was to determine the optimal time for (131)I administration after rhTSH. PARTICIPANTS: Twenty-five euthyroid nongoitrous volunteers participated in the study. DESIGN: Baseline 24-h thyroid (123)I uptake (RAIU) was measured, and then 0.1 mg rhTSH was administered. (123)I was administered 24, 48, or 72 h after rhTSH, and a repeat 24-h RAIU was obtained. SETTING: The study was conducted at an academic research center. MAIN OUTCOME MEASURES: Thyroid function tests, thyroid ultrasounds, and electrocardiograms were measured before rhTSH, then daily for 4 d, and finally 7 d after rhTSH. RESULTS: Serum TSH concentrations 24 h after rhTSH increased from 1.7 +/- 0.5 muU/ml (mean +/- sd) to 13.3 +/- 4. The 24-h RAIUs rose from 25 +/- 5 to 47 +/- 8% (88% increase) when the (123)I was given at 24 h after rhTSH and from 29.8 +/- 7 to 40.5 +/- 13% (36% increase) when the (123)I was given at 48 h and were unchanged when the (123)I was given at 72 h. The post-rhTSH RAIU increase was greater at 24 than at 72 h (P < 0.005) and marginally greater than at 48 h (P = 0.057). Thyroid volumes significantly increased 48 h after rhTSH (10 +/- 3.8 vs. 11.1 +/- 3.7 ml; P < 0.009). Electrocardiograms were normal. CONCLUSIONS: Marked increases in RAIU occurred when (123)I was given 24 h after rhTSH administration to euthyroid volunteers. Smaller increases were observed at 48 h and none at 72 h. PMID- 16317055 TI - Clinical presentation and penetrance of pheochromocytoma/paraganglioma syndromes. AB - CONTEXT: The identification of mutations in genes encoding peptides of succinate dehydrogenase (SDH) in pheochromocytoma/paraganglioma syndromes has necessitated clear elucidation of genotype-phenotype associations. OBJECTIVE: Our objective was to determine genotype-phenotype associations in a cohort of patients with pheochromocytoma/paraganglioma syndromes and succinate dehydrogenase subunit B (SDHB) or subunit D (SDHD) mutations. DESIGN, SETTING, AND PARTICIPANTS: The International SDH Consortium studied 116 individuals (83 affected and 33 clinically unaffected) from 62 families with pheochromocytoma/paraganglioma syndromes and SDHB or SDHD mutations. Clinical data were collected between August 2003 and September 2004 from tertiary referral centers in Australia, France, New Zealand, Germany, United States, Canada, and Scotland. MAIN OUTCOME MEASURES: Data were collected on patients with pheochromocytomas and/or paragangliomas with respect to onset of disease, diagnosis, genetic testing, surgery, pathology, and disease progression. Clinical features were evaluated for evidence of genotype phenotype associations, and penetrance was determined. RESULTS: SDHB mutation carriers were more likely than SDHD mutation carriers to develop extraadrenal pheochromocytomas and malignant disease, whereas SDHD mutation carriers had a greater propensity to develop head and neck paragangliomas and multiple tumors. For the index cases, there was no difference between 43 SDHB and 19 SDHD mutation carriers in the time to first diagnosis (34 vs. 28 yr, respectively; P = 0.3). However, when all mutation carriers were included (n = 112), the estimated age related penetrance was different for SDHB vs. SDHD mutation carriers (P = 0.008). CONCLUSIONS: For clinical follow-up, features of SDHB mutation-associated disease include a later age of onset, extraadrenal (abdominal or thoracic) tumors, and a higher rate of malignancy. In contrast, SDHD mutation carriers, in addition to head and neck paragangliomas, should be observed for multifocal tumors, infrequent malignancy, and the possibility of extraadrenal pheochromocytoma. PMID- 16317056 TI - Transient testicular warming enhances the suppressive effect of testosterone on spermatogenesis in adult cynomolgus monkeys (Macaca fascicularis). AB - CONTEXT: The context of the study was to examine whether combined testosterone (T) and heat (H) treatment have additive or synergistic effects on suppression of spermatogenesis. OBJECTIVE: The objective of the study was to determine whether T+H induces a greater suppression of spermatogenesis than either treatment alone in monkeys. DESIGN: The study was a randomized, placebo-controlled study. SETTING: The study was conducted at a primate center in China. PARTICIPANTS: The study population was comprised of 32 adult cynomolgus monkeys. INTERVENTIONS: Groups of eight adult monkeys were treated for 12 wk with: 1) two empty implants (C); 2) two T implants (T); 3) daily testicular heat exposure (43 C for 30 min) for 2 consecutive days (H); or 4) two T implants plus testicular heat exposure (T+H). Treatment was followed by an 8-wk recovery period. MAIN OUTCOME MEASURES: Measures included sperm counts and germ cell apoptosis. RESULTS: Serum T levels were elevated in both the T and T+H groups during treatment but not in the C or H group. Sperm counts were transiently suppressed after heat to 16.4% of baseline at 4 wk and then returned to pretreatment levels. Sperm counts were suppressed slowly after T treatment to nadir of 6.4% of pretreatment levels at 12 wk. T+H rapidly suppressed sperm output as early as 4 wk to 3.9% of pretreatment levels that was maintained throughout treatment. The decreased sperm counts were due to increased germ cell apoptosis in all treatment groups. Sperm counts recovered to the pretreatment levels in all groups by 8 wk after treatment. CONCLUSION: This proof-of-concept study demonstrates that transient testicular warming enhances and hastens the effect of T implant on the suppression of spermatogenesis in monkeys. PMID- 16317057 TI - Limitation of the homeostasis model assessment to predict insulin resistance and beta-cell dysfunction in older people. AB - CONTEXT: Studies in older people have shown inconsistent agreement between homeostasis model assessment of insulin resistance (HOMA-IR) and dynamic measures of insulin action and have not evaluated HOMA beta-cell. OBJECTIVE: We compared measures of insulin sensitivity and beta-cell function from the frequently sampled iv glucose tolerance test (FSIGT) to HOMA models. DESIGN/PATIENTS/SETTING/INTERVENTION: Two hundred fourteen young and old with normal glucose tolerance (NGT) and old with impaired glucose tolerance (IGT) participated in a retrospective analysis of FSIGT data in a university medical setting. MAIN OUTCOME MEASURE: Sensitivity to insulin (S(I)) and acute insulin response to glucose (AIRg) from FSIGT were compared with HOMA models. RESULTS: S(I) and HOMA-IR measures identified similar patterns of increasing insulin resistance in the two older groups, compared with younger people with NGT, with the greatest degree of insulin resistance in older people with IGT (P < 0.05 vs. young and old NGT for both S(I) and HOMA-IR). Agreement between HOMA-IR and S(I) was moderate (weighted kappa = 0.51). AIRg was similar in young and old NGT but was markedly decreased in old IGT (P < 0.05 vs. young and old NGT). HOMA-beta cell was similar in the three groups. Agreement between HOMA beta-cell and AIRg was weak (weighted kappa = 0.35). CONCLUSIONS: HOMA-IR may detect age-related insulin resistance when comparing large populations of older people. However, dynamic testing appears to be necessary to quantitate diminished insulin secretion in older people. PMID- 16317058 TI - Dihydrotestosterone decreases tumor necrosis factor-alpha and lipopolysaccharide induced inflammatory response in human endothelial cells. AB - CONTEXT: An increasing body of evidence suggests that testosterone may exert beneficial effects on the development of atherosclerosis. It was suggested that testosterone may act after conversion into estradiol and activation of the estrogen receptors; however, a direct role of androgens on the vascular wall has been proposed. OBJECTIVE: We investigated the effects of dihydrotestosterone on the proinflammatory response observed in human endothelial cells. DESIGN: Human endothelial cells isolated from umbilical cords were incubated with lipopolysaccharide or TNFalpha in the presence or absence of dihydrotestosterone (DHT). mRNA and cellular proteins were processed for gene expression studies, and transient transfection experiments were performed to investigate molecular mechanisms involved in the effects observed. SETTING: These studies took place at the Department of Pharmacological Sciences, University of Milan, Milan, Italy. RESULTS: Lipopolysaccharide and TNFalpha induced VCAM-1 and ICAM-1 mRNA and protein expression, as detected by real-time quantitative PCR, fluorescence activated cell sorting, and confocal microscopy, but this effect was inhibited when cells were incubated with DHT. In addition, DHT inhibited mRNA expression of IL-6, MCP-1, CD40, TLR4, PAI-1, and Cox-2 and the release of cytokines and chemokines such as GRO, granulocyte-macrophage colony-stimulating factor, and TNF. The DHT effect was counteracted by bicalutamide, an antagonist of the androgen receptor. Furthermore, when cells were cotransfected with a Cox-2 promoter or a 3X-NF-kappaB luciferase reporter vector and a plasmid expressing the human androgen receptor, DHT treatment inhibited the increase of the luciferase activity observed with TNFalpha. CONCLUSION: DHT could positively regulate endothelial function through the control of the inflammatory response mediated by nuclear factor-kappaB in endothelial cells. PMID- 16317059 TI - Short-term effects of growth hormone on sleep abnormalities in Prader-Willi syndrome. AB - CONTEXT: GH was approved for Prader-Willi Syndrome (PWS) in 2000. Fatalities in individuals with PWS soon after beginning GH treatment prompted concern about GH worsening sleep apnea. OBJECTIVE: We sought to determine whether GH affects sleep apnea in individuals with PWS. DESIGN: Twenty-five patients with PWS had overnight polysomnography (PSA) at baseline and 6 wk after starting GH. SETTING: The study was conducted in a sleep lab using a standardized procedure. PATIENTS: The patients studied had genetically confirmed PWS. MAIN OUTCOME MEASURES: PSA results were analyzed for frequency and severity of central and obstructive apnea/hypopnea events and total apnea/hypopnea index. RESULTS: As a group, GH improved apnea/hypopnea index by a mean of 1.2 events per hour (P = 0.02) and central events by a median of 1.7 events per hour (P < 0.001). Fourteen patients had improvement in obstructive events by a mean of 1.7 events per hour. Six patients had worsening of obstructive events on GH. Four of these patients had upper respiratory tract infections at the time of the second PSA and had tonsil/adenoid hypertrophy on otorhinolaryngological evaluation. Two patients with high serum IGF-I levels had increased obstructive events. CONCLUSIONS: Most of our PWS patients had improvement after short-term GH treatment, but 32% had worsening of sleep disturbance. A subset of PWS patients are at risk during this window of vulnerability shortly after initiation of GH. Because it is difficult to predict who will worsen with GH, patients with PWS should have PSA before and after starting GH and should be monitored for sleep apnea with upper respiratory tract infections. Otorhinolaryngological evaluation is warranted if sleep apnea worsens on GH. IGF-I levels should be monitored, with the goal being physiological levels. PMID- 16317060 TI - Leptin replacement therapy modulates circulating lymphocyte subsets and cytokine responsiveness in severe lipodystrophy. AB - CONTEXT: We conducted this study to understand the role of leptin therapy in immunomodulation. OBJECTIVE: Our objective was to study lymphocyte subpopulations and in vitro peripheral blood mononuclear cell (PBMC) activation during a study evaluating the effects of leptin on metabolic functions in severe lipodystrophy (serum leptin levels < 4 ng/ml). DESIGN AND SETTING: We conducted an open-label study with patients serving as their own control at the Clinical Research Center of the National Institutes of Health. PATIENTS: Ten patients (age range, 15-63 yr; one male and nine females) with generalized forms of lipodystrophy were studied. INTERVENTION: Patients were treated with recombinant human leptin to achieve high normal concentrations for 4 to 8 months. RESULTS: Leptin levels increased from 1.8 +/- 0.4 to 16.5 +/- 3.9 ng/dl (P < 0.001), whereas metabolic control improved [glycosylated hemoglobin (HbA(1c)) fell from 9.3 +/- 0.4 to 7.1 +/- 1.4%, P < 0.001, and triglycerides decreased by 45 +/- 11% from a mean of 1490 +/- 710 mg/dl, P = 0.001]. Lymphocyte subsets were studied by flow cytometry at baseline and at 4 and 8 months of therapy. PBMC responsiveness was evaluated by cytokine release and proliferation after stimulation with phytohemagglutinin, phytohemagglutinin plus IL-12, lipopolysaccharide, and lipopolysaccharide plus interferon-gamma at baseline and 4 months. Various T lymphocyte subsets were significantly lower than age- and sex-matched controls at baseline; however, the CD4/CD8 ratio was normal. The relative percentages of B lymphocytes and monocytes were elevated, although the absolute levels were normal. Leptin therapy induced significant changes in T lymphocyte subsets, which normalized both the absolute number of T lymphocyte subsets and relative percentages of all lineages. Additionally, in vitro TNF-alpha secreted from PBMC of patients was significantly increased to normal after 4 months of leptin therapy compared with baseline. CONCLUSION: These data support existing evidence that leptin has a modest immunomodulatory effect in hypoleptinemic humans. PMID- 16317061 TI - Overweight humans are resistant to the weight-reducing effects of melanocortin4 10. AB - CONTEXT: By enhancing energy expenditure and suppressing appetite, melanocortin peptides derived from proopiomelanocortin play a primary role in the hypothalamic regulation of body weight. In a recent study in normal-weight adults, the 6-wk intranasal administration of the MSH/ACTH(4-10) core fragment of proopiomelanocortin resulted in a distinct reduction of body weight and body fat, accompanied by significant decreases in leptin and insulin plasma concentrations. OBJECTIVE: The present study aimed to generalize this finding to overweight patients. DESIGN, SUBJECTS, AND INTERVENTION: MSH/ACTH(4-10) (0.5 mg) and placebo were intranasally administered once in the morning and once in the evening over a period of 12 wk in 23 overweight men (body mass index, mean +/- sem: 29.72 +/- 0.43 kg/m(2)). RESULTS: MSH/ACTH(4-10) did not induce any significant reduction in body weight, body fat, and plasma levels of insulin and leptin as compared with the effects of placebo. Melanocortin treatment was accompanied by reduced cortisol concentrations. CONCLUSIONS: We conclude that contrasting with normal weight humans, overweight subjects are not susceptible to the effects of melanocortin administration on hypothalamic weight regulatory systems. In overweight subjects, a decreased sensitivity to ACTH/MSH peptides may derive from alterations at the level of the melanocortin receptor or at subsequent steps in the processing of the body fat signal. PMID- 16317062 TI - A new variant in the human Kv1.3 gene is associated with low insulin sensitivity and impaired glucose tolerance. AB - CONTEXT: The voltage-gated potassium channel Kv1.3 (KCNA3) is expressed in a variety of tissues including liver and skeletal muscle. In animal models, knockout of Kv1.3 has been found to improve insulin sensitivity and glucose tolerance. OBJECTIVE: We examined whether mutations in the Kv1.3 gene exist in humans and whether they are associated with alterations of glucose homeostasis. DESIGN AND SETTING: We conducted a genotype-phenotype association study at a university hospital. PARTICIPANTS AND METHODS: In 50 nondiabetic subjects, we screened approximately 4.5 kb of chromosome 1 comprising the single exon, the promoter/5'-untranslated region, and the 3'-untranslated region of the human Kv1.3 gene for mutations by direct sequencing. Subsequently, all identified single-nucleotide polymorphisms were analyzed in 552 nondiabetic subjects who underwent an oral glucose tolerance test (OGTT). Of these, 304 had undergone an additional hyperinsulinemic euglycemic clamp. MAIN OUTCOME MEASURES: We assessed postprandial blood glucose during OGTT and insulin sensitivity measured by hyperinsulinemic euglycemic clamp. RESULTS: We identified five single-nucleotide polymorphisms in the promoter region (T-548C, G-697T, A-845G, T-1645C, and G 2069A) with allelic frequencies of the minor allele of 26, 23, 9, 41, and 16%, respectively. The -1645C allele was associated with higher plasma glucose concentrations in the 2-h OGTT (P = 0.03) even after adjustment for sex, age, and body mass index (P = 0.002). In addition, it was associated with lower insulin sensitivity (P = 0.01, adjusted for sex, age, and body mass index). Functional in vitro analysis using EMSA showed differential transcription factor binding to the T-1645C polymorphism. CONCLUSIONS: We show that a variant in the promoter of the Kv1.3 gene is associated with impaired glucose tolerance and lower insulin sensitivity. Therefore, the Kv1.3 channel represents a candidate gene for type 2 diabetes. PMID- 16317063 TI - Administration of Lispro insulin with meals improves glycemic control, increases circulating leptin, and suppresses ghrelin, compared with regular/NPH insulin in female patients with type 1 diabetes. AB - CONTEXT: Overweight and obesity are overrepresented in adolescents with type 1 diabetes mellitus (T1DM). Exogenous insulin administration often poorly reproduces normal insulin patterns and may less effectively regulate leptin and ghrelin, two hormones involved in the control of appetite and adiposity. OBJECTIVE: The objective of the study was to determine whether insulin regimens that better replicate normal insulin patterns and augment postprandial nutrient disposal may help normalize leptin and ghrelin and improve body weight regulation. DESIGN, SETTING, AND PARTICIPANTS: Ten young women with T1DM were studied in this 2-wk prospective, balanced crossover-design study at the University of California, Davis. INTERVENTION: Participants received either a single injection of regular + NPH insulin (R+N) or two mealtime injections of Lispro insulin in randomized order on 2 separate days. Meal composition and total insulin administered were the same on both treatment days. MAIN OUTCOME MEASURES: Plasma glucose, insulin, leptin, and ghrelin concentrations were monitored over the 10-h study period. RESULTS: Lispro produced two distinct mealtime peaks of insulin, compared with one prolonged rise with R+N. Lispro reduced postprandial hyperglycemia and total glucose area under the curve. Leptin increased more on the Lispro (2.7 +/- 0.7 vs. 0.7 +/- 0.5 ng/ml, P = 0.02). Ghrelin was more suppressed after lunch with Lispro (P = 0.004). CONCLUSIONS: Injection of Lispro insulin with meals produces more physiological insulin patterns, better glucose control, and improved leptin and ghrelin regulation than R+N. More closely mimicking normal insulin, leptin, and ghrelin responses to meals with fast-acting insulin may have implications for body weight regulation in T1DM. PMID- 16317064 TI - An evolving view of the eukaryotic oligosaccharyltransferase. AB - Asparagine-linked glycosylation (ALG) is one of the most common protein modification reactions in eukaryotic cells, as many proteins that are translocated across or integrated into the rough endoplasmic reticulum (RER) carry N-linked oligosaccharides. Although the primary focus of this review will be the structure and function of the eukaryotic oligosaccharyltransferase (OST), key findings provided by the analysis of the archaebacterial and eubacterial OST homologues will be reviewed, particularly those that provide insight into the recognition of donor and acceptor substrates. Selection of the fully assembled donor substrate will be considered in the context of the family of human diseases known as congenital disorders of glycosylation (CDG). The yeast and vertebrate OST are surprisingly complex hetero-oligomeric proteins consisting of seven or eight subunits (Ost1p, Ost2p, Ost3p/Ost6p, Ost4p, Ost5p, Stt3p, Wbp1p, and Swp1p in yeast; ribophorin I, DAD1, N33/IAP, OST4, STT3A/STT3B, Ost48, and ribophorin II in mammals). Recent findings from several laboratories have provided overwhelming evidence that the STT3 subunit is critical for catalytic activity. Here, we will consider the evolution and assembly of the eukaryotic OST in light of recent genomic evidence concerning the subunit composition of the enzyme in diverse eukaryotes. PMID- 16317065 TI - Tetracycline inhibits W7FW14F apomyoglobin fibril extension and keeps the amyloid protein in a pre-fibrillar, highly cytotoxic state. AB - A significant number of fatal diseases are classified as protein deposition disorders, in which a normally soluble protein is deposited in an insoluble amyloid form. It has been reported that tetracycline exhibits anti-amyloidogenic activity by inhibiting aggregate formation and disaggregating preformed fibrils. In this work, we examined the effect induced by the presence of tetracycline on the fibrillogenesis and cytotoxicity of the amyloid-forming apomyoglobin mutant W7FW14F. Like other amyloid-forming proteins, early prefibrillar aggregates formed by this protein are highly cytotoxic, whereas insoluble mature fibrils are not. The effect induced by tetracycline on the fibrillation process has been examined by atomic force microscopy, light scattering, DPH staining, and thioflavin T fluorescence. The cytotoxicity of the amyloid aggregates was estimated by measuring cell viability using MTT assay. The results show that tetracycline acts as anti-aggregating agent, which inhibits the fibril elongation process but not the early aggregation steps leading to the formation of soluble oligomeric aggregates. Thus, this inhibition keeps the W7FW14F mutant in a prefibrillar, highly cytotoxic state. In this respect, a careful usage of tetracycline as fibril inhibitor is indicated. PMID- 16317066 TI - Luminally active, nonabsorbable CFTR inhibitors as potential therapy to reduce intestinal fluid loss in cholera. AB - Enterotoxin-mediated secretory diarrheas such as cholera involve chloride secretion by enterocytes into the intestinal lumen by the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel. We previously identified glycine hydrazide CFTR blockers that by electrophysiological studies appeared to block the CFTR anion pore at its lumen-facing surface. Here, we synthesize highly water-soluble, nonabsorbable malondihydrazides by coupling 2,4 disulfobenzaldehyde, 4-sulfophenylisothiocyante, and polyethylene glycol (PEG) moieties to 2-naphthalenylamino-[(3,5-dibromo-2,4-dihydroxyphenyl) methylene] propanedioic acid dihydrazide, and aminoacethydrazides by coupling PEG to [(N-2 naphthalenyl)-2-(2-hydroxyethyl)]-glycine-2-[(3,5-dibromo-2,4-dihydroxyphenyl) methylene] hydrazide. Compounds rapidly, fully and reversibly blocked CFTR mediated chloride current with Ki of 2-8 microM when added to the apical surface of epithelial cell monolayers. Compounds did not pass across Caco-2 monolayers, and were absorbed by <2%/hr in mouse intestine. Luminally added compounds blocked by >90% cholera toxin-induced fluid secretion in mouse intestinal loops, without inhibiting intestinal fluid absorption. These orally administered, nonabsorbable, nontoxic CFTR inhibitors may reduce intestinal fluid losses in cholera. PMID- 16317067 TI - Allergen-induced peribronchial fibrosis and mucus production mediated by IkappaB kinase beta-dependent genes in airway epithelium. AB - In response to inflammation or injury, airway epithelial cells express inducible genes that may contribute to allergen-induced airway remodeling. To determine the contribution of epithelial cell NF-kappaB activation to the remodeling response, we generated CC10-Cre(tg)/Ikkbeta(delta/delta) mice in which NF-kappaB signaling through IkappaB kinase beta (IKKbeta) is selectively ablated in the airway epithelium by conditional Cre-recombinase expression from the Clara cell (CC10) promoter. Repetitive ovalbumin challenge of mice deficient in airway epithelial IKKbeta prevented nuclear translocation of the RelA NF-kappaB subunit only in airway epithelial cells, resulting in significantly lower peribronchial fibrosis in CC10-Cre(tg)/Ikkbeta(delta/delta) mice compared with littermate controls as assessed by peribronchial trichrome staining and total lung collagen content. Levels of airway mucus, airway eosinophils, and peribronchial CD4+ cells in ovalbumin-challenged mice were also reduced significantly upon airway epithelial Ikkbeta ablation. The diminished inflammatory response was associated with reduced expression of NF-kappaB-regulated chemokines, including eotaxin-1 and thymus- and activation-regulated chemokine, which attract eosinophils and Th2 cells, respectively, into the airway. The number of peribronchial cells expressing TGF-beta1, as well as TGF-beta1 amounts in bronchoalveolar lavage, were also significantly reduced in mice deficient in airway epithelium IKKbeta. Overall, these studies show an important role for NF-kappaB regulated genes in airway epithelium in allergen-induced airway remodeling, including peribronchial fibrosis and mucus production. PMID- 16317068 TI - Endocrine self and gut non-self intersect in the pancreatic lymph nodes. AB - The autoimmune cascade that culminates in diabetes initiates within pancreatic lymph nodes (PLNs). Here, we show that developmentally controlled lymphogenesis establishes a preferential trafficking route from the gut to the PLN, where T cells can be activated by antigens drained from the peritoneum and the gastrointestinal tract. Furthermore, intestinal stress modifies the presentation of pancreatic self-antigens in PLNs. The convergence of endocrine and intestinal contents within PLNs has significant implications for type 1 diabetes and may help to explain the link between autoimmune pathogenesis and environmental provocation. PMID- 16317069 TI - Profiling condition-specific, genome-wide regulation of mRNA stability in yeast. AB - The steady-state abundance of an mRNA is determined by the balance between transcription and decay. Although regulation of transcription has been well studied both experimentally and computationally, regulation of transcript stability has received little attention. We developed an algorithm, MatrixREDUCE, that discovers the position-specific affinity matrices for unknown RNA-binding factors and infers their condition-specific activities, using only genomic sequence data and steady-state mRNA expression data as input. We identified and computationally characterized the binding sites for six mRNA stability regulators in Saccharomyces cerevisiae, which include two members of the Pumilio-homology domain (Puf) family of RNA-binding proteins, Puf3p and Puf4p. We provide computational and experimental evidence that regulation of mRNA stability by these factors is modulated in response to a variety of environmental stimuli. PMID- 16317070 TI - Application of compression-based distance measures to protein sequence classification: a methodological study. AB - MOTIVATION: Distance measures built on the notion of text compression have been used for the comparison and classification of entire genomes and mitochondrial genomes. The present study was undertaken in order to explore their utility in the classification of protein sequences. RESULTS: We constructed compression based distance measures (CBMs) using the Lempel-Zlv and the PPMZ compression algorithms and compared their performance with that of the Smith-Waterman algorithm and BLAST, using nearest neighbour or support vector machine classification schemes. The datasets included a subset of the SCOP protein structure database to test distant protein similarities, a 3-phosphoglycerate kinase sequences selected from archaean, bacterial and eukaryotic species as well as low and high-complexity sequence segments of the human proteome, CBMs values show a dependence on the length and the complexity of the sequences compared. In classification tasks CBMs performed especially well on distantly related proteins where the performance of a combined measure, constructed from a CBM and a BLAST score, approached or even slightly exceeded that of the Smith-Waterman algorithm and two hidden Markov model-based algorithms. PMID- 16317071 TI - SimShift: identifying structural similarities from NMR chemical shifts. AB - MOTIVATION: An important quantity that arises in NMR spectroscopy experiments is the chemical shift. The interpretation of these data is mostly done by human experts; to our knowledge there are no algorithms that predict protein structure from chemical shift sequences alone. One approach to facilitate this process could be to compare two such sequences, where the structure of one protein has already been resolved. Our claim is that similarity of chemical shifts thereby found implies structural similarity of the respective proteins. RESULTS: We present an algorithm to identify structural similarities of proteins by aligning their associated chemical shift sequences. To evaluate the correctness of our predictions, we propose a benchmark set of protein pairs that have high structural similarity, but low sequence similarity (because with high sequence similarity the structural similarities could easily be detected by a sequence alignment algorithm). We compare our results with those of HHsearch and SSEA and show that our method outperforms both in >50% of all cases. PMID- 16317072 TI - Comparison of Bayesian and maximum-likelihood inference of population genetic parameters. AB - Comparison of the performance and accuracy of different inference methods, such as maximum likelihood (ML) and Bayesian inference, is difficult because the inference methods are implemented in different programs, often written by different authors. Both methods were implemented in the program MIGRATE, that estimates population genetic parameters, such as population sizes and migration rates, using coalescence theory. Both inference methods use the same Markov chain Monte Carlo algorithm and differ from each other in only two aspects: parameter proposal distribution and maximization of the likelihood function. Using simulated datasets, the Bayesian method generally fares better than the ML approach in accuracy and coverage, although for some values the two approaches are equal in performance. MOTIVATION: The Markov chain Monte Carlo-based ML framework can fail on sparse data and can deliver non-conservative support intervals. A Bayesian framework with appropriate prior distribution is able to remedy some of these problems. RESULTS: The program MIGRATE was extended to allow not only for ML(-) maximum likelihood estimation of population genetics parameters but also for using a Bayesian framework. Comparisons between the Bayesian approach and the ML approach are facilitated because both modes estimate the same parameters under the same population model and assumptions. PMID- 16317073 TI - Identification of plant microRNA homologs. AB - MicroRNAs (miRNAs) are a recently discovered class of non-coding RNAs that regulate gene and protein expression in plants and animals. MiRNAs have so far been identified mostly by specific cloning of small RNA molecules, complemented by computational methods. We present a computational identification approach that is able to identify candidate miRNA homologs in any set of sequences, given a query miRNA. The approach is based on a sequence similarity search step followed by a set of structural filters. PMID- 16317074 TI - Application of latent semantic analysis to protein remote homology detection. AB - MOTIVATION: Remote homology detection between protein sequences is a central problem in computational biology. The discriminative method such as the support vector machine (SVM) is one of the most effective methods. Many of the SVM-based methods focus on finding useful representations of protein sequence, using either explicit feature vector representations or kernel functions. Such representations may suffer from the peaking phenomenon in many machine-learning methods because the features are usually very large and noise data may be introduced. Based on these observations, this research focuses on feature extraction and efficient representation of protein vectors for SVM protein classification. RESULTS: In this study, a latent semantic analysis (LSA) model, which is an efficient feature extraction technique from natural language processing, has been introduced in protein remote homology detection. Several basic building blocks of protein sequences have been investigated as the 'words' of 'protein sequence language', including N-grams, patterns and motifs. Each protein sequence is taken as a 'document' that is composed of bags-of-word. The word-document matrix is constructed first. The LSA is performed on the matrix to produce the latent semantic representation vectors of protein sequences, leading to noise-removal and smart description of protein sequences. The latent semantic representation vectors are then evaluated by SVM. The method is tested on the SCOP 1.53 database. The results show that the LSA model significantly improves the performance of remote homology detection in comparison with the basic formalisms. Furthermore, the performance of this method is comparable with that of the complex kernel methods such as SVM-LA and better than that of other sequence based methods such as PSI-BLAST and SVM-pairwise. PMID- 16317075 TI - Conservation analysis of large biochemical networks. AB - MOTIVATION: Large biochemical networks pose a unique challenge from the point of view of evaluating conservation laws. The computational problem in most cases exceeds the capability of available software tools, often resulting in inaccurate computation of the number and form of conserved cycles. Such errors have profound effects on subsequent calculations, particularly in the evaluation of the Jacobian which is a critical quantity in many other calculations. The goal of this paper is to outline a new algorithm that is computationally efficient and robust at extracting the correct conservation laws for very large biochemical networks. RESULTS: We show that our algorithm can perform the conservation analysis of large biochemical networks, and can evaluate the correct conserved cycles when compared with other similar software tools. Biochemical simulators such as Jarnac and COPASI are successful at extracting only a subset of the conservation laws that our algorithm can. This is illustrated with examples for some large networks which show the advantages of our method. PMID- 16317076 TI - Systems Biology Toolbox for MATLAB: a computational platform for research in systems biology. AB - We present a Systems Biology Toolbox for the widely used general purpose mathematical software MATLAB. The toolbox offers systems biologists an open and extensible environment, in which to explore ideas, prototype and share new algorithms, and build applications for the analysis and simulation of biological and biochemical systems. Additionally it is well suited for educational purposes. The toolbox supports the Systems Biology Markup Language (SBML) by providing an interface for import and export of SBML models. In this way the toolbox connects nicely to other SBML-enabled modelling packages. Models are represented in an internal model format and can be described either by entering ordinary differential equations or, more intuitively, by entering biochemical reaction equations. The toolbox contains a large number of analysis methods, such as deterministic and stochastic simulation, parameter estimation, network identification, parameter sensitivity analysis and bifurcation analysis. PMID- 16317077 TI - Paircoil2: improved prediction of coiled coils from sequence. AB - We introduce Paircoil2, a new version of the Paircoil program, which uses pairwise residue probabilities to detect coiled-coil motifs in protein sequence data. Paircoil2 achieves 98% sensitivity and 97% specificity on known coiled coils in leave-family-out cross-validation. It also shows superior performance compared with published methods in tests on proteins of known structure. PMID- 16317078 TI - Effects of a reduction in maternal renal mass on pregnancy and cardiovascular and renal function of the pregnant ewe. AB - Maternal renal disease is associated with high maternal and fetal morbidity. To establish an animal model to study renal dysfunction in pregnancy and its potential role in programming for renal disease and hypertension in adult life, a kidney was removed from each of 16 nonpregnant ewes, and a branch of the renal artery of the remaining kidney was ligated (STNx ewes). The 16 STNx and 15 intact ewes were time mated 2.5-17 mo later and studied at 119-132 days of gestation. STNx ewes demonstrated renal hypertrophy and glomerular hyperfiltration. They had higher diastolic arterial pressures (P < 0.05) and larger left ventricles (P < 0.0005), drank more water (P < 0.01), were hypochloremic (P < 0.01) and hyperglycemic (P < 0.0005), and had higher plasma creatinine levels (P < 0.0005) than intact ewes. Effective renal plasma flows and glomerular filtration rates were lower (P < 0.01) and protein excretion was greater (P < 0.05) in STNx than in intact ewes. Glomerulotubular balance was impaired in STNx ewes. Proximal tubular Na(+) reabsorption was reduced (P < 0.05), so Na(+) excretion was increased (P < 0.05). In STNx ewes, filtered K(+) loads were reduced (P < 0.005), but K(+) excretion was the same as in intact ewes. There was net K(+) secretion in STNx ewes; in intact ewes, there was net reabsorption. Plasma renin and angiotensinogen concentrations in STNx and intact ewes were similar, so the hypertension in STNx ewes was not renin dependent. STNx fetuses grew normally, and their blood gases, blood pressure, and heart rates were normal. These alterations in maternal fluid and electrolyte balance and the potential risk of maternal salt depletion or hyperkalemia may adversely affect the fetus. PMID- 16317079 TI - Identification of novel transcriptional networks in response to treatment with the anticarcinogen 3H-1,2-dithiole-3-thione. AB - 3H-1,2-dithiole-3-thione (D3T), an inducer of antioxidant and phase 2 genes, is known to enhance the detoxification of environmental carcinogens, prevent neoplasia, and elicit other protective effects. However, a comprehensive view of the regulatory pathways induced by this compound has not yet been elaborated. Fischer F344 rats were gavaged daily for 5 days with vehicle or D3T (0.3 mmol/kg). The global changes of gene expression in liver were measured with Affymetrix RG-U34A chips. With the use of functional class scoring, a semi supervised method exploring both the expression pattern and the functional annotation of the genes, the Gene Ontology classes were ranked according to the significance of the impact of D3T treatment. Two unexpected functional classes were identified for the D3T treatment, cytosolic ribosome constituents with 90% of those genes increased, and cholesterol biosynthesis with 91% of the genes repressed. In another novel approach, the differentially expressed genes were evaluated by the Ingenuity computational pathway analysis tool to identify specific regulatory networks and canonical pathways responsive to D3T treatment. In addition to the known glutathione metabolism pathway (P = 0.0011), several other significant pathways were also revealed, including antigen presentation (P = 0.000476), androgen/estrogen biosynthesis (P = 0.000551), fatty acid (P = 0.000216), and tryptophan metabolism (P = 0.000331) pathways. These findings showed a profound impact of D3T on lipid metabolism and anti-inflammatory/immune suppressive response, indicating a broader cytoprotective effect of this compound than previously expected. PMID- 16317080 TI - Identification and characterization of a novel member of the ATP-sensitive K+ channel subunit family, Kir6.3, in zebrafish. AB - ATP-sensitive K+ (KATP) channels play a crucial role in coupling cellular metabolism to membrane potential. In addition to the orthologs corresponding to Kir6.1 and Kir6.2 of mammals, we have identified a novel member, designated Kir6.3 (zKir6.3), of the inward rectifier K+ channel subfamily Kir6.x in zebrafish. zKir6.3 is a protein of 432 amino acids that shares 66% identity with mammalian Kir6.2 but differs considerably from mammalian Kir6.1 and Kir6.2 in the COOH terminus, which contain an Arg-Lys-Arg (RKR) motif, an endoplasmic reticulum (ER) retention signal. Single-channel recordings of reconstituted channels show that zKir6.3 requires the sulfonylurea receptor 1 (SUR1) subunit to produce KATP channel currents with single-channel conductance of 57.5 pS. Confocal microscopic analysis shows that zebrafish Kir6.3 requires the SUR1 subunit for its trafficking to the plasma membrane. Analyses of chimeric protein between human Kir6.2 and zKir6.3 and a COOH-terminal deletion of zKir6.3 indicate that interaction between the COOH terminus of zKir6.3 and SUR1 is critical for both channel activity and trafficking to the plasma membrane. We also identified zebrafish orthologs corresponding to mammalian SUR1 (zSUR1) and SUR2 (zSUR2) by the genomic database. Both Kir6.3 and SUR1 are expressed in embryonic brain of zebrafish, as assessed by whole mount in situ hybridization. These data indicate that Kir6.3 and SUR1 form functional KATP channels at the plasma membrane in zebrafish through a mechanism independent from ER retention by the RKR motif. PMID- 16317081 TI - High-affinity peptide transporter PEPT2 (SLC15A2) of the zebrafish Danio rerio: functional properties, genomic organization, and expression analysis. AB - Solute carrier 15 (SLC15) membrane proteins PEPT1 (SLC15A1) and PEPT2 (SLC15A2) have been described in great detail in mammals. In contrast, information in lower vertebrates is limited. We characterized the functional properties of a novel zebrafish peptide transporter orthologous to mammalian and avian PEPT2, described its gene (pept2) structure, and determined mRNA tissue distribution. An expressed sequence tag (EST) cDNA (Integrated Molecular Analysis of Gene Expression; IMAGE) corresponding to zebrafish pept2 was completed by inserting a stretch of 75 missing nucleotides in the coding sequence to obtain a 3,238-bp functional clone. The complete open reading frame (ORF) was 2,160 bp and encoded a 719-amino acid protein. Electrophysiological analysis after cRNA injection in Xenopus laevis oocytes suggested that zebrafish PEPT2 is a high-affinity/low-capacity transporter (K(0.5) for glycyl-L-glutamine approximately 18 microM at -120 mV and pH 7.5). Zebrafish pept2 gene was 19,435 kb, thus being the shortest vertebrate pept2 fully characterized so far. Also, zebrafish pept2 exhibited 23 exons and 22 introns, whereas human and rodent pept2 genes contain 22 exons and 21 introns only. Zebrafish pept2 mRNA was mainly detected in brain, kidney, gut, and, interestingly, otic vesicle, the embryonic structure that develops into the auditory/vestibular organ, homolog to the higher vertebrate inner ear, of the adult fish. Characterization of zebrafish pept2 will contribute to the investigation of peptide transporters using a well-established genetic model and will allow the elucidation of the evolutionary and functional relationships among vertebrate peptide transporters. Moreover, it can represent a useful marker to screen mutations that affect choroid plexus and inner ear development. PMID- 16317082 TI - Divergence between motoneurons: gene expression profiling provides a molecular characterization of functionally discrete somatic and autonomic motoneurons. AB - Studies in the developing spinal cord suggest that different motoneuron (MN) cell types express very different genetic programs, but the degree to which adult programs differ is unknown. To compare genetic programs between adult MN columnar cell types, we used laser capture microdissection (LCM) and Affymetrix microarrays to create expression profiles for three columnar cell types: lateral and medial MNs from lumbar segments and sympathetic preganglionic motoneurons located in the thoracic intermediolateral nucleus. A comparison of the three expression profiles indicated that approximately 7% (813/11,552) of the genes showed significant differences in their expression levels. The largest differences were observed between sympathetic preganglionic MNs and the lateral motor column, with 6% (706/11,552) of the genes being differentially expressed. Significant differences in expression were observed for 1.8% (207/11,552) of the genes when comparing sympathetic preganglionic MNs with the medial motor column. Lateral and medial MNs showed the least divergence, with 1.3% (150/11,552) of the genes being differentially expressed. These data indicate that the amount of divergence in expression profiles between identified columnar MNs does not strictly correlate with divergence of function as defined by innervation patterns (somatic/muscle vs. autonomic/viscera). Classification of the differentially expressed genes with regard to function showed that they underpin all fundamental cell systems and processes, although most differentially expressed genes encode proteins involved in signal transduction. Mining the expression profiles to examine transcription factors essential for MN development suggested that many of the same transcription factors participate in combinatorial codes in embryonic and adult neurons, but patterns of expression change significantly. PMID- 16317083 TI - AT1 receptors are necessary for eccentric training-induced hypertrophy and strength gains in rat skeletal muscle. AB - This study was undertaken to measure the response of skeletal muscle to eccentric contractions (EC) in the presence of the angiotensin type 1 (AT1) receptor blocker, losartan. It was hypothesized that blocking AT1 receptors prior to an initial bout of EC would prevent the muscle from developing the normal adaptation to EC as demonstrated by the repeated bout effect. It was also hypothesized that continuous AT1 receptor blockade during EC training would significantly reduce muscle hypertrophy and strength gains that occur with repeated EC. Rats received losartan in their drinking water at either a low dose (20 mg (kg body weight)-1 day-1) or a high dose (40 mg (kg body weight)-1 day-1). Each bout of EC consisted of a total of 24 contractions. Rats were assigned to four groups: a single acute bout of EC (n=6); two bouts of EC separated by 14 days (n=8); and 4 weeks of training twice a week on the low dose (n=5) or the high dose (n=9). There was no effect of AT1 receptor blockade on the initial loss of function following a single acute bout of EC, or on the repeated bout effect following a second exposure to EC. AT1 receptor blockade did alter the results of EC training, in both the low and high dose groups. Losartan treatments prevented EC training induced increases in muscle wet and dry weights compared to untreated rats. Finally, the low and high dose losartan treatments also prevented an increase in muscle contractile force following EC training compared to the untreated group. Functional AT1 receptors are therefore not necessary for an acute adaptation to EC as demonstrated by the repeated bout effect, but are necessary for muscle hypertrophy and increased contractile force associated with EC training. PMID- 16317084 TI - Transient NMDA receptor-mediated hypoperfusion following umbilical cord occlusion in preterm fetal sheep. AB - Exposure to severe hypoxia leads to delayed cerebral and peripheral hypoperfusion. There is evidence in the very immature brain that transient abnormal glutaminergic receptor activity can occur during this phase of recovery. We therefore examined the role of N-methyl-D-aspartate (NMDA) receptor activity in mediating secondary hypoperfusion in preterm fetal sheep at 70% of gestation. Fetuses received either sham asphyxia or asphyxia and were studied for 12 h recovery. The specific, non-competitive NMDA receptor antagonist dizocilpine maleate (2 mg kg-1 bolus plus 0.07 mg kg h-1i.v.) or saline (vehicle) was infused from 15 min after asphyxia until 4 h. In the asphyxia-vehicle group abnormal epileptiform EEG transients were observed during the first 4 h of reperfusion, the peak of which corresponded approximately to the nadir in peripheral and cerebral hypoperfusion. Dizocilpine significantly suppressed this activity (2.7+/ 1.3 versus 11.2+/-2.7 counts min-1 at peak frequency, P<0.05) and markedly delayed and attenuated the rise in vascular resistance in both peripheral and cerebral vascular beds observed after asphyxia, effectively preventing the initial deep period of hypoperfusion in carotid blood flow and femoral blood flow (P<0.01). However, while continued infusion did attenuate subsequent transient tachycardia, it did not prevent the development of a secondary phase of persistent but less profound hypoperfusion. In conclusion, the present studies suggest that in the immature brain the initial phase of delayed cerebral and peripheral hypoperfusion following exposure to severe hypoxia is mediated by NMDA receptor activity. The timing of this effect in the cerebral circulation corresponds closely to abnormal EEG activity, suggesting a pathological glutaminergic activation that we speculate is related to evolving brain injury. PMID- 16317085 TI - The unfolded protein response: a novel component of the hypoxic stress response in tumors. AB - Hypoxia is a physiologically important endoplasmic reticulum (ER) stress that is present in all solid tumors. Numerous clinical studies have shown that tumor hypoxia predicts for decreased local control, increased distant metastases, and decreased overall survival in a variety of human tumors. Hypoxia selects for tumors with an increased malignant phenotype and increases the metastatic potential of tumor cells. Tumor cells respond to hypoxia and ER stress through the activation of the unfolded protein response (UPR). The UPR is an adaptive response to increase cell survival during ER stress. XBP-1 is a critical transcriptional regulator of this process and is required for tumor growth. Pancreatic ER kinase (PKR-like ER kinase) regulates the translational branch of the UPR and is also important in the growth of tumors. Although the exact mechanism has yet to be elucidated, recent data suggest that the UPR affects tumor growth through protection from apoptosis and may influence angiogenic signaling pathways. Targeting various components of the UPR is a promising therapeutic strategy. Understanding the relationship between hypoxia, the UPR, and tumor growth is crucial to improving current cancer therapies. PMID- 16317086 TI - Nicotine induces cyclooxygenase-2 and vascular endothelial growth factor receptor 2 in association with tumor-associated invasion and angiogenesis in gastric cancer. AB - Blockade of angiogenesis is a promising strategy to suppress tumor growth, invasion, and metastasis. Vascular endothelial growth factor (VEGF), which binds to tyrosine kinase receptors [VEGF receptors (VEGFR) 1 and 2], is the mediator of angiogenesis and mitogen for endothelial cells. Cyclooxygenase-2 (COX-2) plays an important role in the promoting action of nicotine on gastric cancer growth. However, the action of nicotine and the relationship between COX-2 and VEGF/VEGFR system in tumorigenesis remain undefined. In this study, the effects of nicotine in tumor angiogenesis, invasiveness, and metastasis were studied with sponge implantation and Matrigel membrane models. Nicotine (200 microg/mL) stimulated gastric cancer cell proliferation, which was blocked by SC-236 (a highly selective COX-2 inhibitor) and CBO-P11 (a VEGFR inhibitor). This was associated with decreased VEGF levels as well as VEGFR-2 but not VEGFR-1 expression. Topical injection of nicotine enhanced tumor-associated vascularization, with a concomitant increase in VEGF levels in sponge implants. Again, application of SC 236 (2 mg/kg) and CBO-P11 (0.4 mg/kg) partially attenuated vascularization by approximately 30%. Furthermore, nicotine enhanced tumor cell invasion through the Matrigel membrane by 4-fold and promoted migration of human umbilical vein endothelial cells in a cocultured system with gastric cancer cells. The activity of matrix metalloproteinases 2 and 9 and protein expressions of plasminogen activators (urokinase-type plasminogen activator and its receptor), which are the indicators of invasion and migration processes, were increased by nicotine but blocked by COX-2 and VEGFR inhibitors. Taken together, our results reveal that the promoting action of nicotine on angiogenesis, tumor invasion, and metastasis is COX-2/VEGF/VEGFR dependent. PMID- 16317087 TI - Both hypomethylation and hypermethylation in a 0.2-kb region of a DNA repeat in cancer. AB - NBL2 is a tandem 1.4-kb DNA repeat, whose hypomethylation in hepatocellular carcinomas was shown previously to be an independent predictor of disease progression. Here, we examined methylation of all cytosine residues in a 0.2-kb subregion of NBL2 in ovarian carcinomas, Wilms' tumors, and diverse control tissues by hairpin-bisulfite PCR. This new genomic sequencing method detects 5 methylcytosine on covalently linked complementary strands of a DNA fragment. All DNA clones from normal somatic tissues displayed symmetrical methylation at seven CpG positions and no methylation or only hemimethylation at two others. Unexpectedly, 56% of cancer DNA clones had decreased methylation at some normally methylated CpG sites as well as increased methylation at one or both of the normally unmethylated sites. All 146 DNA clones from 10 cancers could be distinguished from all 91 somatic control clones by assessing methylation changes at three of these CpG sites. The special involvement of DNA methyltransferase 3B in NBL2 methylation was indicated by analysis of cells from immunodeficiency, centromeric region instability, and facial anomalies syndrome patients who have mutations in the gene encoding DNA methyltransferase 3B. Blot hybridization of 33 cancer DNAs digested with CpG methylation-sensitive enzymes confirmed that NBL2 arrays are unusually susceptible to cancer-linked hypermethylation and hypomethylation, consistent with our novel genomic sequencing findings. The combined Southern blot and genomic sequencing data indicate that some of the cancer-linked alterations in CpG methylation are occurring with considerable sequence specificity. NBL2 is an attractive candidate for an epigenetic cancer marker and for elucidating the nature of epigenetic changes in cancer. PMID- 16317088 TI - p53-independent regulation of p21Waf1/Cip1 expression and senescence by Chk2. AB - The Chk2 kinase is a tumor suppressor and key component of the DNA damage checkpoint response that encompasses cell cycle arrest, apoptosis, and DNA repair. It has also been shown to have a role in replicative senescence resulting from dysfunctional telomeres. Some of these functions are at least partially exerted through activation of the p53 transcription factor. High-level expression of virally transduced Chk2 in A549 human lung carcinoma cells led to arrested proliferation, apoptosis, and senescence. These were accompanied by various molecular events, including p21(Waf1/Cip1) (p21) transcriptional induction, consistent with p53 activation. However, Chk2-dependent senescence and p21 transcriptional induction also occurred in p53-defective SK-BR-3 (breast carcinoma) and HaCaT (immortalized keratinocyte) cells. Small interfering RNA mediated knockdown of p21 in p53-defective cells expressing Chk2 resulted in a decrease in senescent cells. These results revealed a p53-independent role for Chk2 in p21 induction and senescence that may contribute to tumor suppression and genotoxic treatment outcome. PMID- 16317089 TI - Mammalian target of rapamycin promotes vincristine resistance through multiple mechanisms independent of maintained glycolytic rate. AB - Deregulation of the phosphoinositide 3-kinase-Akt pathway is a major contributor to oncogenesis and resistance to cancer therapy. Recent work has shown mammalian target of rapamycin (mTOR) to be a major target downstream of Akt that contributes to both transformation and therapeutic resistance. Although inhibitors of Akt are not yet clinically available, rapamycin, a mTOR-specific inhibitor, has long been used as an immunosuppressant, and several rapamycin analogues are now in clinical trials in oncology. Recent data indicate that a mTOR complex phosphorylates Akt, and this complex is insensitive to rapamycin. We show that dominant-negative mTOR diminishes phosphorylation of endogenous Akt and exogenous myristoylated Akt (mAkt), that prolonged exposure to rapamycin also inhibits Akt activation, and that this inhibition is dependent on new protein synthesis. These data suggest that mTOR facilitates Akt activation through mechanisms other than direct phosphorylation. A constitutively active mTOR mutant that fails to enhance Akt phosphorylation nevertheless promotes resistance to multiple antimicrotubule agents, indicating that mTOR also mediates survival independent of Akt. Although Akt- and mTOR-mediated survival has been linked to regulation of cellular metabolism, we also show that survival and metabolic control are separable. The hexokinase inhibitor 5-thioglucose markedly inhibits glycolytic rate but does not diminish vincristine resistance mediated by mAkt or mTOR, and it has only a minor effect on mTOR- or mAkt-mediated resistance to growth factor withdrawal, suggesting that Akt-mTOR-mediated resistance is largely independent of maintenance of glycolytic rate. We conclude that mTOR activity can promote resistance through multiple mechanisms independent of maintained glycolytic rate. PMID- 16317090 TI - Delta-like1-induced Notch1 signaling regulates the human plasmacytoid dendritic cell versus T-cell lineage decision through control of GATA-3 and Spi-B. AB - Human early thymic precursors have the potential to differentiate into multiple cell lineages, including T cells and plasmacytoid dendritic cells (pDCs). This decision is guided by the induction or silencing of lineage-specific transcription factors. The ETS family member Spi-B is a key regulator of pDC development, whereas T-cell development is critically dependent on GATA-3. Here we show that triggering of the Notch1 signaling pathway by Delta-like1 controls the T/pDC lineage decision by regulating the balance between these factors. CD34+ CD1a- thymic progenitor cells express Notch1, but down-regulate this receptor when differentiating into pDCs. On coculture with stromal cell lines expressing either human Delta-like1 (DL1) or Jagged1 (Jag1) Notch ligands, thymic precursors express GATA-3 and develop into CD4+ CD8+ TCRalphabeta+ T cells. On the other hand, DL1, but not Jag1, down-regulates Spi-B expression, resulting in impaired development of pDCs. The Notch1-induced block in pDC development can be relieved through the ectopic expression of Spi-B. These data indicate that DL1-induced activation of the Notch1 pathway controls the lineage commitment of early thymic precursors by altering the levels between Spi-B and GATA-3. PMID- 16317091 TI - Migration inhibitory factor up-regulates vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 via Src, PI3 kinase, and NFkappaB. AB - Cell adhesion molecules are critical in monocyte (MN) recruitment in immune mediated and hematologic diseases. We investigated the novel role of recombinant human migration inhibitory factor (rhMIF) in up-regulating vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) and their signaling pathways in human MNs. rhMIF-induced expression of VCAM-1 and ICAM-1 was significantly higher compared with nonstimulated MNs. rhMIF induced MN VCAM-1 and ICAM-1 expression in a concentration-dependent manner (P < .05). Antisense oligodeoxynucleotides (ODNs) and inhibitors of Src, PI3K, p38, and NFkappaB significantly reduced rhMIF-induced MN VCAM-1 and ICAM-1 expression (P < .05). However, Erk1/2 and Jak2 were not involved. Silencing RNA directed against MIF, and inhibitors of Src, PI3K, NFkappaB, anti-VCAM-1, and anti-ICAM-1 significantly inhibited rhMIF-induced adhesion of HL-60 cells to human dermal microvascular endothelial cells (HMVECs) or an endothelial cell line, HMEC-1, in cell adhesion assays, suggesting the functional significance of MIF-induced adhesion molecules (P < .05). rhMIF also activated MN phospho-Src, -Akt, and -NFkappaB in a time dependent manner. rhMIF induced VCAM-1 and ICAM-1 up-regulation in 12 hours via Src, PI3K, and NFkappaB as shown by Western blotting and immunofluorescence. MIF and MIF-dependent signaling pathways may be a potential target for treating diseases characterized by up-regulation of cell adhesion molecules. PMID- 16317092 TI - Impaired class switch recombination (CSR) in Waldenstrom macroglobulinemia (WM) despite apparently normal CSR machinery. AB - Analysis of clonotypic isotype class switching (CSR) in Waldenstrom macroglobulinemia (WM) and IgM monoclonal gammopathy of undetermined significance (MGUS) reveals a normal initial phase of B-cell activation as determined by constitutive and inducible expression of activation-induced cytidine deaminase (AID). Switch mu (Smu) analysis shows that large deletions are not common in WM or IgM MGUS. In CD40L/IL-4-stimulated WM cultures from 2 patients, we observed clonotypic IgG exhibiting intraclonal homogeneity associated with multiple hybrid Smu/Sgamma junctions. This suggests CSR had occurred within WM cells. Nevertheless, the estimated IgG/IgM-cell frequency was relatively low (1/1600 cells). Thus, for the majority of WM B cells, CSR does not occur even when stimulated in vitro, suggesting that the WM cell is constitutively unable to or being prevented from carrying out CSR. In contrast to WM, the majority of IgM MGUS clones exhibit intraclonal heterogeneity of IgH VDJ. Furthermore, most IgM MGUS accumulate more mutations in the upstream Smu region than do WM, making them unlikely WM progenitors. These observations suggest that switch sequence analysis may identify the subset of patients with IgM MGUS who are at risk of progression to WM. PMID- 16317093 TI - De novo synthesis of early growth response factor-1 is required for the full responsiveness of mast cells to produce TNF and IL-13 by IgE and antigen stimulation. AB - Early growth-response factor 1 (Egr-1) is a zinc-finger transcription factor that plays a regulatory role in the expression of many genes important for inflammation. Whether Egr-1 is involved in IgE-dependent mast-cell activation was investigated. We demonstrated that IgE and antigen (TNP) stimulation induced a rapid expression of Egr-1 mRNA in mouse bone marrow-derived mast cells (BMMCs). As early as 15 to 20 minutes after IgE + TNP stimulation, Egr-1 protein was detectable in the nucleus of BMMCs by immunofluorescence or electrophoretic mobility shift assay. To examine a role for Egr-1 in IgE-dependent cytokine production by mast cells, Egr-1-deficient (Egr-1-/-) BMMCs were developed from the bone marrow cells of Egr-1 knockout mice. Egr-1-/- BMMCs express similar levels of surface c-kit and IgE receptor as compared with those on Egr-1+/+ BMMCs. Importantly, IgE + TNP-induced TNF and IL-13 expression was significantly reduced at both mRNA and protein levels in Egr-1-/- BMMCs as compared with those in Egr-1+/+ BMMCs. Thus, our results suggest that de novo synthesis of Egr-1 represents a novel mechanism in FcepsilonRI signaling and is required for the full responsiveness of IgE-dependent TNF and IL-13 production by mast cells. PMID- 16317094 TI - Graft rejection after unrelated donor hematopoietic stem cell transplantation for thalassemia is associated with nonpermissive HLA-DPB1 disparity in host-versus graft direction. AB - The success of allogeneic hematopoietic stem cell transplantation (HSCT) from matched unrelated donors (UDs) for beta-thalassemia may be hampered by the occurrence of graft rejection. Here, we show that the rate of this complication can be reduced by selecting 5-loci HLA-matched donors without nonpermissive mismatches at HLA-DPB1, defined according to an algorithm previously described and based on principles of central T-cell tolerance. Seventy-two consecutive patients and their UDs, prospectively selected for matching at the allelic level for HLA-A, -B, -C, -DRB, and -DQB1 loci, were enrolled in the analysis. These pairs were either DPB1 matched/permissively mismatched (n = 45, control group) or had at least one nonpermissive DPB1 mismatch in the host-versus-graft (HvG; n = 17) or in the graft-versus-host (GvH; n = 10) direction. In multivariate analysis, the risk of rejection was significantly increased in the group with HvG disparity (RR = 7.42; 95% CI = 1.29-42.68; P = .02) as compared to the control group. A lower, statistically significant, probability of thalassemia-free survival was found in patients belonging to the HvG group as compared to controls (RR = 5.15; 95% CI = 1.58-16.82; P = .01). These data suggest that in patients with thalassemia, the incidence of graft failure after HSCT may be reduced by appropriate selection of UDs, with such selection taking into account the functional rules of immunogenetics. PMID- 16317095 TI - The extracellular adherence protein (Eap) of Staphylococcus aureus inhibits wound healing by interfering with host defense and repair mechanisms. AB - Staphylococcus aureus is a major human pathogen interfering with host-cell functions. Impaired wound healing is often observed in S aureus-infected wounds, yet, the underlying mechanisms are poorly defined. Here, we identify the extracellular adherence protein (Eap) of S aureus to be responsible for impaired wound healing. In a mouse wound-healing model wound closure was inhibited in the presence of wild-type S aureus and this effect was reversible when the wounds were incubated with an isogenic Eap-deficient strain. Isolated Eap also delayed wound closure. In the presence of Eap, recruitment of inflammatory cells to the wound site as well as neovascularization of the wound were prevented. In vitro, Eap significantly reduced intercellular adhesion molecule 1 (ICAM-1)-dependent leukocyte-endothelial interactions and diminished the consequent activation of the proinflammatory transcription factor nuclear factor kappaB (NFkappaB) in leukocytes associated with a decrease in expression of tissue factor. Moreover, Eap blocked alphav-integrin-mediated endothelial-cell migration and capillary tube formation, and neovascularization in matrigels in vivo. Collectively, the potent anti-inflammatory and antiangiogenic properties of Eap provide an underlying mechanism that may explain the impaired wound healing in S aureus infected wounds. Eap may also serve as a lead compound for new anti-inflammatory and antiangiogenic therapies in several pathologies. PMID- 16317096 TI - Upon viral exposure, myeloid and plasmacytoid dendritic cells produce 3 waves of distinct chemokines to recruit immune effectors. AB - Host response to viral infection involves distinct effectors of innate and adaptive immunity, whose mobilization needs to be coordinated to ensure protection. Here we show that influenza virus triggers, in human blood dendritic cell (DC) subsets (ie, plasmacytoid and myeloid DCs), a coordinated chemokine (CK) secretion program with 3 successive waves. The first one, occurring at early time points (2 to 4 hours), includes CKs potentially attracting effector cells such as neutrophils, cytotoxic T cells, and natural killer (NK) cells (CXCL16, CXCL1, CXCL2, and CXCL3). The second one occurs within 8 to 12 hours and includes CKs attracting effector memory T cells (CXCL8, CCL3, CCL4, CCL5, CXCL9, CXCL10, and CXCL11). The third wave, which occurs after 24 to 48 hours, when DCs have reached the lymphoid organs, includes CCL19, CCL22, and CXCL13, which attract naive T and B lymphocytes. Thus, human blood DC subsets carry a common program of CK production, which allows for a coordinated attraction of the different immune effectors in response to viral infection. PMID- 16317097 TI - Array comparative genomic hybridization reveals genomic copy number changes associated with outcome in diffuse large B-cell lymphomas. AB - To identify, in high-resolution regions of DNA, the copy number changes associated with outcome in patients with diffuse large B-cell lymphoma (DLBCL), a disease with an approximately 50% mortality rate, we performed array comparative genomic hybridization (array-CGH) on specimens from 64 patients with newly diagnosed DLBCL treated with anthracycline-based chemotherapy. For the entire cohort, 55 commonly gained/lost regions, ranging in size from less than 1 Mbp to entire chromosomes, were identified using 1- to 2-Mbp and 2- to 4-Mbp resolution BAC arrays. Copy number changes of 9 minimal regions significantly correlated with overall survival, of which 6 were 10 Mbp or smaller. On multivariate analysis, loss of chromosomes 2 (2.4-4.1 Mbp) and 16 (33.8-35.6 Mbp) were found to be prognostic indicators of poor survival, independent of clinical features routinely used to predict outcome. Loss of chromosome 1 (78.2-79.1 Mbp) was predictive of good outcome. For a subset of 55 specimens classified according to cell-of-origin expression signature subtype, gain of chromosome 12 (45.4-53.8 Mbp) was found to be significantly associated with the germinal center B-cell like DLBCL subtype. Overall, array-CGH identified relatively small genomic regions associated with outcome, which, along with follow-up expression studies, may reveal target genes important in DLBCL clinical behavior. PMID- 16317098 TI - Expression of rearranged TCRgamma genes in natural killer cells suggests a minor thymus-dependent pathway of lineage commitment. AB - Natural killer (NK) cells are thought to develop from common lymphoid progenitors in the bone marrow. However, immature thymocytes also retain NK potential. Currently, the contribution of the thymus-dependent pathway in normal steady state NK-cell development is unknown. Here, we show that TCRgamma genes are rearranged in approximately 5% of neonatal and 1% of adult mouse splenic NK cells, and similar levels are detected in NK cells from TCRbeta,delta double knockout mice, excluding the possibility of T-cell contamination. NK-cell TCRgamma gene rearrangement is thymus dependent because this rearrangement is undetectable in nude mouse NK cells. These results change the current view of NK cell development and show that a subset of NK cells develops from immature thymocytes that have rearranged TCRgamma genes. PMID- 16317099 TI - A novel murine model of fetal and neonatal alloimmune thrombocytopenia: response to intravenous IgG therapy. AB - Fetal and neonatal alloimmune thrombo cytopenia (FNAITP) is a life-threatening bleeding disorder caused by maternal antibodies directed against fetal platelet antigens. The immunoreactive epitopes in FNAITP are primarily located in the extracellular regions of the platelet glycoprotein IIIa (beta3 integrin). Here we have established a novel animal model of FNAITP using beta3 integrin-deficient (beta3-/-) mice. We demonstrated first that these mice are immunoresponsive to beta3 integrin; beta3-/- mice transfused with wild-type platelets generated specific anti-beta3 antibodies which were able to induce thrombocytopenia in wild type mice. Subsequently, beta3-/- female mice (both naive and immunized) were bred with wild-type male mice to recapitulate the features of FNAITP. The titer of generated maternal antibodies correlated with the severity of FNAITP. High titer maternal anti-beta3 anti-bodies caused severe fetal thrombocytopenia, intracranial hemorrhage, and even miscarriage. Furthermore, maternal administration of intravenous immunoglobulin G (IgG) ameliorated FNAITP and down regulated pathogenic antibodies in both the maternal and fetal circulations. PMID- 16317100 TI - Circulating thrombopoietin as an in vivo growth factor for blast cells in acute myeloid leukemia. AB - Thrombopoietin (TPO), the major growth factor for cells of the megakaryocytic lineage, is removed from circulation by binding to c-mpl receptors present on platelets and megakaryocytes. We studied patients with acute lymphoblastic leukemia (ALL) or acute myeloblastic leukemia (AML) and used TPO-induced c-fos protein up-regulation as a marker of c-mpl functionality and observed that c-mpl presenting blast cells were present in 62% (37 of 60) of patients with ALL but that c-mpl was nonfunctional in 0 of 28 patients and that they were present in 56% (22 of 39) of patients with AML and were functional in 43% (12 of 28). Adequate increases in serum TPO level in response to thrombocytopenia were seen in patients with ALL and with c-mpl-deficient (c-mpl-) AML. In contrast, in patients with c-mpl-proficient (c-mpl+) AML, TPO levels were found to be inappropriately low but increased to expected values during induction chemotherapy as blasts disappeared. In vitro significant TPO-associated blast cell proliferation or decreased apoptosis was observed only in patients with c mpl+ AML compared with ALL or c-mpl- AML and was highly correlated with low in vivo TPO levels (P < .001). These data suggest that, in patients with AML, inadequate TPO levels are secondary to TPO clearing by functional c-mpl receptor myeloid blast cells and that TPO may serve as an in vivo myeloid leukemic growth factor in a significant number of patients. PMID- 16317101 TI - Mast cells and neutrophils proteolytically activate chemokine precursor CTAP-III and are subject to counterregulation by PF-4 through inhibition of chymase and cathepsin G. AB - The CXC chemokines platelet factor 4 (PF-4/CXCL4) and connective tissue activating peptide III (CTAP-III) are released by activated human platelets in micromolar concentrations. So far, neutrophils have been recognized to cleave the precursor CTAP-III to form the active chemokine neutrophil-activating peptide 2 (NAP-2/CXCL7) through limited proteolysis by membrane-associated cathepsin G. Here we show for the first time that activated human skin mast cells (MCs) convert CTAP-III into biologically active NAP-2 through proteolytic cleavage by released chymase. A direct comparison on a cell number basis revealed that unstimulated MCs exceed the CTAP-III-processing potency of neutrophils about 30 fold, whereas MCs activated by IgE cross-linking exhibit even 1000-fold higher CTAP-III-processing capacity than fMLP-stimulated neutrophils. Intriguingly, PF-4 counteracted MC- as well as neutrophil-mediated NAP-2 generation at physiologically relevant concentrations. Addressing the underlying mechanism, we obtained evidence that PF-4 acts as an inhibitor of the CTAP-III-processing enzymes cathepsin G and chymase without becoming cleaved itself as a competitive substrate. Because cleavage of the CTAP-III-unrelated substrate substance P was also affected by PF-4, our results suggest a regulatory role for PF-4 not only in NAP-2 generation but also in neutrophil- and MC-mediated processing of other physiologically relevant inflammatory mediators. PMID- 16317102 TI - SLAM/SLAM interactions inhibit CD40-induced production of inflammatory cytokines in monocyte-derived dendritic cells. AB - Signaling lymphocyte activation molecule (SLAM, CD150, or SLAMF1) is a self ligand receptor on the surface of activated T- and B-lymphocytes, macrophages, and dendritic cells (DCs). Here we examine the effect of SLAM/SLAM interactions on CD40L-induced CD40 signaling pathways in human DCs. CD40L-expressing L929 cells induced DCs to produce interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and IL-12, which was strongly inhibited by coexpression of SLAM on the surface of the L929 cells. Similarly, transfection of DCs with SLAM strongly reduced CD40L-induced IL-12 production. Furthermore, the negative effect of SLAM/SLAM interactions on CD40L-induced DC activation was also detected in the presence of lipopolysaccharide (LPS). LPS-induced IL-12 secretion, however, was not inhibited by SLAM engagement. CD40L-activated DCs affected by exposure to SLAM/SLAM engagement were impaired in their ability to induce differentiation of naive T lymphocytes into interferon-gamma (IFN-gamma)-producing T-helper 1 (Th1) effector cells. These inhibitory effects were not the result of a general unresponsiveness of DCs to CD40L, as SLAM/SLAM interactions did not prevent CD40L induced up-regulation of CD83, CD86, or human leukocyte antigen (HLA)-DQ on the surface of DCs. Taken together, the results indicate that SLAM/SLAM interactions inhibit CD40-induced signal transduction in monocyte-derived dendritic cells, an effect that was not detectable in earlier studies using anti-SLAM monoclonal antibodies. PMID- 16317103 TI - Strikingly homologous immunoglobulin gene rearrangements and poor outcome in VH3 21-using chronic lymphocytic leukemia patients independent of geographic origin and mutational status. AB - We recently reported that Swedish VH3-21-using chronic lymphocytic leukemia (CLL) patients showed restricted immunoglobulin gene features and poor prognosis despite VH mutation status. To investigate this further, we analyzed the VH and VL gene rearrangements in 90 VH3-21+ patients from Sweden, Germany, Italy, United States, Finland, and Australia and correlated these data with survival and other prognostic markers. Sixty-three percent exhibited mutated VH genes and 37% unmutated VH genes. Fifty (56%) patients displayed a short and homologous heavy chain CDR3, many of these with the amino acid motif DANGMDV. Also, a highly biased Vlambda2-14 use was evident in 72% of patients with a restricted light chain CDR3, QVWDS(S/G)SDHPWV. Combined restricted heavy- and light-chain CDR3s were found in patients from all included countries. Although VH3-21+ CLLs have a remarkably predominant lambda expression, analyses of kappa deleting element indicated a conserved light-chain rearrangement order. The overall survival was poor in the VH3-21+ cohort (median survival, 88 months), with no significant difference in relation to mutation status or CDR3 homology. High ZAP-70 and CD38 expression was found in both mutated and unmutated VH3-21+ cases as well as a slight increase of 11q-aberrations. In summary, highly restricted B-cell receptors and worse outcome characterize VH3-21+ CLLs independent of geographic origin and mutation status. PMID- 16317104 TI - EBV latency III immortalization program sensitizes B cells to induction of CD95 mediated apoptosis via LMP1: role of NF-kappaB, STAT1, and p53. AB - Epstein-Barr virus (EBV) induces CD95 expression and the CD95 gene (FAS) is regulated by NF-kappaB, STAT1, and/or p53. To understand the contribution of these factors in the regulation of CD95 by EBV in lymphoblastoid cell lines (LCLs), we cloned dominant-active IkappaBalpha, active (STAT1alpha) and inactive (STAT1beta) forms of STAT1, p53, a dominant-negative mutant of LMP1, and wild type LMP1 into a novel double-inducible episomal vector, pRT-1. These plasmids were stably transfected either into wild-type LCLs or EREB2-5 cells, an LCL with an estrogen-regulatable EBNA2 protein. Inhibition of LMP1 signaling decreased expression of CD95, whereas overexpression of LMP1 markedly increased it. Induction of the latency III program in EREB2-5 cells correlated with activation of NF-kappaB, STAT1, and p53. CD95 expression was regulated by these 3 transcriptional systems. STAT1 and p53 activation were secondary to NF-kappaB activation. CD95 surface expression sensitized EBV-infected B cells to the induction of CD95-mediated apoptosis. In vitro inhibition of CD95-CD95 ligand interaction was found to reverse T-cell killing of EBV-infected B cells. Therefore, LMP1 activation of NF-kappaB sensitizes infected B cells to CD95 mediated apoptosis and renders EBV latency III-immortalized B cells susceptible to elimination by the immune system, contributing to the establishment of a host/virus equilibrium. PMID- 16317105 TI - Neoplastic circulating endothelial cells in multiple myeloma with 13q14 deletion. AB - In multiple myeloma (MM), circulating endothelial cells (CECs) represent a vascular marker of angiogenesis and may reflect tumor mass. In this report, we showed that, in 5 MM patients with 13q14 deletion, CECs carried the same chromosome aberration as the neoplastic plasma cells (11%-32% of CECs with 13q14 deletion). Most of the CECs displayed immunophenotypic features of endothelial progenitor cells as they expressed CD133, a marker gradually lost during endothelial differentiation and absent on mature endothelial cells. To the contrary, in 3 patients with monoclonal gammopathy of undetermined significance and 13q14 deletion, CECs were cytogenetically normal and had a mature immunophenotype. In MM CECs, immunoglobulin genes were clonally rearranged. These findings suggest a possible origin of CECs from a common hemangioblast precursor that can give rise to both plasma cells and endothelial cells and point to a direct contribution of MM-derived CECs to tumor vasculogenesis and possibly to the spreading and progression of the disease. PMID- 16317106 TI - Blood pressure and heart rate effects following a single dose of bitter orange. AB - BACKGROUND: The ingredients of numerous "ephedra-free" dietary supplements used for weight loss include bitter orange, which contains sympathomimetic alkaloids such as synephrine. Due to the similarity in chemical structure to ephedrine and the potential sympathomimetic effects of synephrine, it is hypothesized that bitter orange may increase blood pressure (BP) and heart rate (HR). OBJECTIVE: To determine the effects on BP and HR after a single dose of bitter orange in healthy adults. METHODS: In a prospective, randomized, double-blind, placebo controlled, crossover study, 15 young, healthy, adult subjects received either a single dose of Nature's Way Bitter Orange--a 900 mg dietary supplement extract standardized to 6% synephrine--or matching placebo, with a one week washout period. Systolic BP (SBP), diastolic BP (DBP), and HR were measured at baseline and every hour for 6 hours after administration. RESULTS: SBP after bitter orange was significantly increased versus placebo at hours 1-5 (p < 0.0001); the peak difference was 7.3 +/- 4.6 mm Hg. Although the baseline DBP was higher than after administration of both placebo and bitter orange, DBP after bitter orange was significantly increased versus placebo at hours 4 and 5 (p < or = 0.02); the peak difference was 2.6 +/- 3.8 mm Hg. HR was significantly increased after bitter orange versus placebo for hours 2-5 (p < 0.01); the peak difference was 4.2 +/- 4.5 beats/min. CONCLUSIONS: SBP, DBP, and HR were higher for up to 5 hours after a single dose of bitter orange versus placebo in young, healthy adults. PMID- 16317107 TI - Hepatotoxicity associated with acarbose therapy. AB - OBJECTIVE: To report a case of acarbose-induced hepatotoxicity and compare other reported cases from the literature. CASE SUMMARY: A 57-year-old woman with type 2 diabetes mellitus for about 10 years had been treated with insulin glargine 20 units/day since December 19, 2002. Acarbose 100 mg 3-times-daily add-on therapy for inadequate glycemic control was started on June 5, 2003. Six months later, the woman complained of gastrointestinal discomfort; the acarbose dose was decreased to 50 mg 3 times daily thereafter. Laboratory examination later revealed alanine aminotransferase (ALT) 640 U/L (upper reference value 55). To elucidate the possibilities of adverse reactions caused by concurrent use of nutritional supplements and medication, we discontinued propolis extract, Ginkgo biloba, placeta extract, and estrogen. Although no remarkable symptoms were noted thereafter, the abnormal ALT values persisted, and no definite viral or autoimmune etiologies were identified. Acarbose was discontinued in August 2004; aspartate aminotransferase and ALT values returned to normal in October 2004. DISCUSSION: In addition to ruling out other possible etiologic factors, we assessed the probability of acarbose-induced hepatotoxicity by observing the close time relationship between drug administration and the development of signs and symptoms, as well as the close time relationship between drug withdrawal and the normalization of abnormal liver function test values. An objective causality assessment revealed that an adverse drug reaction was probable as determined by both the Naranjo probability scale and the Roussel Uclaf Causality Assessment Method score. CONCLUSIONS: Although acarbose-induced hepatotoxicity appears to be uncommon, diabetic patients receiving long-term acarbose therapy should be closely monitored for this adverse effect. PMID- 16317108 TI - Determinants of the differences in LDL-cholesterol after initiation of statin treatment. AB - BACKGROUND: Randomized clinical trials may not accurately predict drug benefit in clinical practice because the patients and conditions of therapy almost always differ between those settings. OBJECTIVE: To determine whether differences in low density lipoprotein cholesterol (LDL-C) time curves between clinical trial results and usual care experience after initiation of statin therapy could be explained fully in terms of observable characteristics of patients and practice in usual care. METHODS: We compared LDL-C measurements for 3 years after initiation of statin treatment in individuals enrolled in a health maintenance organization (HMO), with enrollees in the active treatment arm of the CARE (Cholesterol and Recurrent Events) trial. Analysis of the determinants of variation in LDL-C in the HMO cohort was used to adjust the crude results to the distribution of patient and treatment characteristics in the trial. RESULTS: The mean percent decrease in LDL-C was lower in the HMO cohort (n = 1245) than in the clinical trial at the end of each 6-month period, with the difference diminishing over time. Adjustment of the HMO time curve to the baseline characteristics of the clinical trial cohort did not significantly change the mean estimates at any time point. Assuming optimal adherence in the HMO cohort raised the curve over time, with 95% confidence intervals including the means observed in CARE after 18 months. Fixing treatment to pravastatin 40 mg/day, as in CARE, brought the estimates in the HMO cohort very close to those of the clinical trial, with 95% confidence intervals including the means observed in CARE at all time points. CONCLUSIONS: Treatment selection, dosing, and adherence under usual care were the primary reasons for which improvements in LDL-C in practice fell short of expectations that are based on clinical trial findings for statin therapy. Beginning with a low dose of statins and titrating to a satisfactory response can delay effective treatment by 18 months or more. Poor adherence accounts for a further substantial shortfall from maximal effect. Differences between trial populations and the general population of statin users with respect to age, gender, and baseline LDL-C have no measurable impact on discrepancies between predicted and observed LDL-C improvement in usual practice. PMID- 16317109 TI - 1,4-Diazabicyclo[2.2.2]octane derivatives: a novel class of voltage-gated potassium channel blockers. AB - Voltage-gated potassium (Kv) channels are targets for therapeutic drugs in the treatment of pathologic conditions including cardiac arrhythmia and epilepsy. In this study, we synthesized three classes of novel polyammonium compounds incorporating the bicyclic unit 1,4-diazabicyclo[2.2.2]octane (DABCO) and tested their action on three representative mammalian Kv channels (Kv2.1, Kv3.4, and Kv4.2) expressed in Xenopus laevis oocytes. Nonsubstituted DABCO did not block the Kv channels tested. Simple DABCO monostrings and diDABCO strings inhibited Kv2.1 and Kv3.4 channels, with potency increasing with string length for both these DABCO classes. Both Kv2.1 and Kv3.4 were most sensitive to C16 monostrings, with IC50 values of 1.9 and 0.6 microM, respectively. For compounds comprising two DABCO groups separated by an aromatic ring, inhibition depended upon relative positioning of the two DABCO groups, and only the para form (JC638.2alpha) was active, blocking Kv2.1 with an IC50 of 186 microM. Kv4.2 channels were relatively insensitive to all compounds tested. Unlike the tetraethylammonium ion (TEA), neither JC638.2alpha nor C16 monostring TA279 produced block when applied intracellularly via the recording electrode to Kv2.1 channels expressed in Chinese hamster ovary cells, suggesting against an internal site of action. However, JC638.2alpha protected an introduced cysteine (K356C) in the Kv2.1 outer pore from permanent modification by methanethiosulfonate ethyltrimethylammonium (MTSET). These data suggest that JC638.2alpha occupies an external binding site similar to that of TEA in the Kv2.1 outer pore, but with much higher affinity than TEA. These DABCO salts represent a new class of Kv channel blockers, some with higher potencies than any previously described quaternary ammonium ions. The potential for synthesis of an array of modular derivatives suggests that DABCO compounds hold promise as probes of Kv channel structure and identity and as potential therapeutic agents. PMID- 16317110 TI - Increased divalent metal transporter 1 expression might be associated with the neurotoxicity of L-DOPA. AB - Based on the available data, we speculated that changes in brain iron metabolism induced by L-DOPA might be associated with the neurotoxicity of L-DOPA. To investigate this possibility, the effects of L-DOPA on the expression of iron influx proteins [transferrin receptor (TfR) and divalent metal transporter 1 (DMT1)], iron efflux protein (ferroportin 1), and iron uptake in C6 glioma cells were determined in this study using Northern blot and Western blot analysis and the calcein method. The findings showed that treatment of C6 cells with different concentrations of L-DOPA (0-100 microM) did not affect the expression of mRNA and protein of TfR and DMT1 with iron-responsive element (+IRE) and protein of ferroportin 1. However, a significant increase in the expression of DMT1(-IRE) mRNA and protein was found in cells treated, respectively, with 10 and 30 microM L-DOPA (mRNA) and 1, 5, 10 and 30 microM L-DOPA (protein). The increase in DMT( IRE) protein induced by L-DOPA treatment was in parallel with the increase in DMT(-IRE) mRNA. The levels of DMT1(-IRE) mRNA and protein peaked in the cells treated with 10 microM L-DOPA and then decreased progressively with increasing concentrations of L-DOPA. Further study demonstrated that treatment of the cells with 10 microM L-DOPA induced a significant increase in ferrous uptake by C6 glioma cells. The findings suggested that the increased DMT1(-IRE) expression might be partly associated with the neurotoxicity of L-DOPA. Clinical relevance of the findings needs to be investigated further. PMID- 16317111 TI - The role of Rho-associated kinase in differential regulation by statins of interleukin-1beta- and lipopolysaccharide-mediated nuclear factor kappaB activation and inducible nitric-oxide synthase gene expression in vascular smooth muscle cells. AB - An optimal level of NO has protective effects in atherosclerosis, whereas large amounts contribute to septic shock. To study how statins, the potent inhibitors of cholesterol synthesis, regulate NO in the vascular wall, we determined their effects on interleukin-1beta (IL-1beta)- and lipopolysaccharide (LPS)-induced NO production in aortic vascular smooth muscle cells (VSMCs). Compared with the large amounts of NO and inducible NO synthase (iNOS) protein expression induced by LPS, the responses of IL-1beta were modest. Various statins were found to inhibit LPS-induced iNOS expression and NO production, although they potentiated IL-1beta responses. In addition, fluvastatin increased IL-1beta-induced p65 nuclear translocation and nuclear factor kappaB (NF-kappaB) activity, although it inhibited those induced by LPS. To address the role of small G proteins in statin's actions, farnesyl transferase inhibitors [alpha-hydroxyfarne sylphosphonic acid and (2S)-2-[[(2S)-2-[(2S,3S)-2-[(2R)-2-amino-3 mercaptopropyl]amino]-3-methylpentyl]oxy]-1-oxo-3-phenylpropyl]amino]-4 (methylsulfonyl)-butanoic acid 1-methylethyl ester (L-744382)], Rac inhibitor (NSC23766), and Rho-associated kinase (ROCK) inhibitor [N-(4-pyridyl)-4-(1 aminoethyl)cyclohexanecarboxamide dihydrochloride (Y-27632)] were used. We found that Y-27632 potentiated IL-1beta-induced iNOS expression, p65 nuclear translocation, IkappaB kinase (IKK), and NF-kappaB activation, whereas it had minimal effects on LPS-induced responses. In contrast, farnesyl transferase inhibitors blocked iNOS protein expression induced by LPS and IL-1beta, whereas NSC23766 had no effect. Further studies showed that LPS down-regulated Rho and ROCK activity, whereas IL-1beta increased them, suggesting a negative role of Rho and ROCK signaling, which is regulated in contrary manners by IL-1beta and LPS, in IKK/NF-kappaB activation. Through abrogating this negative signaling, statins differentially regulate iNOS expression induced by LPS and IL-1beta in VSMCs. These differential actions of statins on iNOS gene regulation might provide an additional explanation for the pleiotropic beneficial effects of statins. PMID- 16317112 TI - Involvement of cAMP/cAMP-dependent protein kinase signaling pathway in regulation of Na+,K+-ATPase upon activation of opioid receptors by morphine. AB - The depolarization of neurons induced by impairment of Na+,K+-ATPase activity after long-term opiate treatment has been shown to involve the development of opioid dependence. However, the mechanisms underlying changes in Na+,K+-ATPase activity after opioid treatment are unclear. The best-established molecular adaptation to long-term opioid exposure is up-regulation of the cAMP/cAMP dependent protein kinase (PKA) signaling pathway; this study, therefore, was undertaken to investigate the role of up-regulation of cAMP/PKA signaling pathway in alteration of the mouse hippocampal Na+,K+-ATPase activity. The results demonstrated that short-term morphine treatment dose dependently stimulated Na+,K+-ATPase activity. This action could be significantly suppressed by adenylyl cyclase activator 7beta-acetoxy-8,13-epoxy-1alpha,6beta,9alpha-trihydroxylabd-14 en-11-one (forskolin), or the cAMP analog dibutyryl-cAMP. Contrary to short-term morphine treatment, long-term treatment significantly inhibited Na+,K+-ATPase activity. Moreover, an additional decrease in Na+,K+-ATPase activity was observed by naloxone precipitation. The effects of both short- and long-term morphine treatment on Na+,K+-ATPase activity were naltrexone-reversible. The regulation of Na+,K+-ATPase activity by morphine was inversely correlated with intracellular cAMP accumulation. N-[2-(4-Bromocinnamylamino)ethyl]-5-isoquinoline (H89), a specific PKA inhibitor, mimicked the stimulatory effect of short-term morphine but antagonized the inhibitory effect of long-term morphine treatment on Na+,K+ ATPase activity. However, okadaic acid, a protein phosphatase inhibitor, suppressed short-term morphine stimulation but potentiated long-term morphine inhibition of Na+,K+-ATPase activity. The regulation of Na+,K+-ATPase activity by morphine treatment seemed to associate with the alteration in phosphorylation level but not to be relevant to the change in abundance of Na+,K+-ATPase. These findings strongly demonstrate that cAMP/PKA signaling pathway involves regulation of Na+,K+-ATPase activity after activation of opioid receptors. PMID- 16317113 TI - The amino terminus and the third extracellular loop of CX3CR1 contain determinants critical for distinct receptor functions. AB - The G protein-coupled receptor CX3CR1 is a specific receptor for the CX3C chemokine fractalkine (CX3CL1 according to the new chemokine nomenclature). The aim of this study was to identify receptor elements that contribute independently to agonist binding and receptor activation. Targeted mutation of selected acidic amino acid residues demonstrated that the binding activity of CX3CR1 was critically dependent on the two negatively charged residues Asp25 and Glu254 located on the N-terminal domain and third extracellular loop, respectively. In addition, mutation of the uncharged polar residue Tyr14 in the amino terminus caused a reduction in the ligand binding affinity. In contrast, the three acidic residues Glu13, Asp16, and Asp266 did not contribute to ligand binding but were crucial for receptor activation. The mutant receptors E13A, D16A, and D266A bound fractalkine with high affinity but were unable to induce signaling events necessary to support chemotaxis. These acidic residues may engage in electrostatic interactions with basic residues on fractalkine that are necessary for receptor function but not for binding. Our data are consistent with a model of chemokine receptor activation consisting of a multi-step mechanism. Step one mediates the high-affinity fractalkine binding involving Tyr14, Asp25, and Glu254. The initial interaction then triggers the engagement of Glu13, Asp16, and Asp266, which are necessary for CX3CR1 activation. PMID- 16317114 TI - The hexosamine signaling pathway: deciphering the "O-GlcNAc code". AB - A dynamic cycle of addition and removal of O-linked N-acetylglucosamine (O GlcNAc) at serine and threonine residues is emerging as a key regulator of nuclear and cytoplasmic protein activity. Like phosphorylation, protein O GlcNAcylation dramatically alters the posttranslational fate and function of target proteins. Indeed, O-GlcNAcylation may compete with phosphorylation for certain Ser/Thr target sites. Like kinases and phosphatases, the enzymes of O GlcNAc metabolism are highly compartmentalized and regulated. Yet, O-GlcNAc addition is subject to an additional and unique level of metabolic control. O GlcNAc transfer is the terminal step in a "hexosamine signaling pathway" (HSP). In the HSP, levels of uridine 5'-diphosphate (UDP)-GlcNAc respond to nutrient excess to activate O-GlcNAcylation. Removal of O-GlcNAc may also be under similar metabolic regulation. Differentially targeted isoforms of the enzymes of O-GlcNAc metabolism allow the participation of O-GlcNAc in diverse intracellular functions. O-GlcNAc addition and removal are key to histone remodeling, transcription, proliferation, apoptosis, and proteasomal degradation. This nutrient-responsive signaling pathway also modulates important cellular pathways, including the insulin signaling cascade in animals and the gibberellin signaling pathway in plants. Alterations in O-GlcNAc metabolism are associated with various human diseases including diabetes mellitus and neurodegeneration. This review will focus on current approaches to deciphering the "O-GlcNAc code" in order to elucidate how O-GlcNAc participates in its diverse functions. This ongoing effort requires analysis of the enzymes of O-GlcNAc metabolism, their many targets, and how the O-GlcNAc modification may be regulated. PMID- 16317115 TI - The influence of dairy product consumption on body composition. AB - Recent epidemiologic research suggests that dairy product intake or its components (calcium, vitamin D, and amount or source of protein) are associated with lower body weight or body fat. Clinical intervention trials designed to test this association during weight loss are promising, but still controversial. Few data are available on the effect of calcium or dairy products on prevention of weight gain in long-term trials. The mechanisms proposed to mediate the putative effect of dietary calcium are primarily the formation of fecal fatty acid complexes to reduce fat absorption and the regulation of energy metabolism, including lipolysis from adipocytes and fatty acid oxidation, through the calciotropic hormones, parathyroid hormone, and 1,25-dihydroxyvitamin D. Increased energy expenditure, increased satiety, or a shift from fat to lean mass must accompany these changes in lipid metabolism to achieve changes in fat mass; however, measurable changes in these other parameters either have not been tested or have not been noted uniformly. If dairy products or their components have an effect on altering fat mass, it is likely to be a small change that may have a substantial effect on the incidence of obesity over time. PMID- 16317116 TI - The VSL# 3 probiotic mixture modifies microflora but does not heal chronic dextran-sodium sulfate-induced colitis or reinforce the mucus barrier in mice. AB - The mucus layer covering the epithelium is one of the main lines of defense of the colonic barrier. Both mucus gel and mucin expressions are altered during colonic inflammation and could be involved in epithelial repair. We postulated that modulating colonic mucus and mucins by probiotic supplementation could contribute to healing inflammatory mucosa. Our aim in this study was to determine whether probiotics could repair dextran-sodium sulfate (DSS)-induced chronic colitis in mice, and whether modifications of the colonic mucins could be involved. For that purpose, the VSL#3 probiotic mixture of 8 lactic acid bacteria probiotic strains was administered daily for 2 wk to mice with a mucosa impaired by a mild DSS treatment, and to mice with a normal mucosa. Probiotic strains survived in the gastrointestinal tract, increased the cecal concentrations of bifidobacteria, and modified cecal microflora metabolic activity in both DSS treated and healthy mice. However, probiotic supplementation did not reverse the inflammation induced by DSS at either the macroscopic or histological level. Concurrently, probiotics did not modify the colonic mucus barrier, in terms of either mucin gene expression or adherent mucus layer thickness. In conclusion, the modification of microflora by supplementation with the VSL#3 probiotic mixture did not help to repair the colonic barrier breakdown caused by DSS treatment. The potential healing roles of mucins were neither confirmed nor invalidated by this study. PMID- 16317117 TI - Copper exposure induces trafficking of the menkes protein in intestinal epithelium of ATP7A transgenic mice. AB - The final steps in the absorption and excretion of copper at the molecular level are accomplished by 2 closely related proteins that catalyze the ATP-dependent transport of copper across the plasma membrane. These proteins, ATP7A and ATP7B, are encoded by the genes affected in human genetic copper-transport disorders, namely, Menkes and Wilson diseases. We studied the effect of copper perfusion of an isolated segment of the jejunum of ATP7A transgenic mice on the intracellular distribution of ATP7A by immunofluorescence of frozen sections. Our results indicate that ATP7A is retained in the trans-Golgi network under copper-limiting conditions, but relocalized to a vesicular compartment adjacent to the basolateral membrane in intestines perfused with copper. The findings support the hypothesis that the basolateral transport of copper from the enterocyte into the portal blood may involve ATP7A pumping copper into a vesicular compartment followed by exocytosis to release the copper, rather than direct pumping of copper across the basolateral membrane. PMID- 16317118 TI - Resveratrol inhibits TNF-alpha-induced proliferation and matrix metalloproteinase expression in human vascular smooth muscle cells. AB - Resveratrol (RV), a polyphenolic substance found in grape skin, was suggested to play a role in preventing the development of atherosclerotic disease. Although RV has antiatherogenic effects on vascular smooth muscle cells (VSMC), the molecular mechanisms associated with tumor necrosis factor (TNF)-alpha-induced VSMC are unclear. The goal of this study was to determine the effect of RV on the modulation of cell proliferation, cell-cycle regulation, and matrix metalloproteinase (MMP)-9 expression in TNF-alpha-induced human VSMC. RV treatment inhibited DNA synthesis in cultured VSMC in the presence of TNF-alpha. These inhibitory effects were associated with reduced levels of extracellular signal-regulated kinase (ERK) 1/2 activity and G(1) cell-cycle arrest. Treatment with RV, which blocks the cell cycle in the G(1) phase, downregulated the expression of cyclins and cyclin-dependent kinases (CDKs) and upregulated the expression of p21/WAF1, a CDK inhibitor. RV did not upregulate p27. Moreover, RV increased the promoter activity of the p21/WAF1 gene. Immunoblot and deletion analysis of the p21/WAF1 promoter showed that RV induced the expression of p21/WAF1 and that this expression was independent of the p53 pathway. Furthermore, zymographic and immunoblot analyses showed that RV dose dependently suppressed the TNF-alpha-induced expression of MMP-9. This inhibition was characterized by the downregulation of MMP-9, which was transcriptionally regulated at the activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) sites in the MMP-9 promoter. Collectively, these results suggest that RV inhibits cell proliferation, G(1) to S phase cell-cycle progress, and MMP-9 expression through the transcription factors NF-kappaB and AP-1 in TNF-alpha-induced VSMC. PMID- 16317119 TI - Vitamin A status in mice affects the histone code of the phosphoenolpyruvate carboxykinase gene in liver. AB - Vitamin A deficiency decreases hepatic phosphoenolpyruvate carboxykinase (PEPCK) gene expression in mice, and expression is restored with retinoic acid (RA) treatment in vivo. In the studies reported here, we examined changes in histone modification and coregulator association with the regulatory domains of the PEPCK gene in response to alterations in vitamin A status. We identified nuclear receptors that bind to retinoic acid response elements (RAREs) in the PEPCK promoter by electrophoretic mobility shift assay and verified these in vivo using chromatin immunoprecipitation in mouse liver. Hypothetically, nuclear receptors at PEPCK RAREs recruit specific coactivator molecules that contribute to the acetylation of core histones and/or serve as bridging molecules between nuclear receptors and basal transcription factors at the transcription start site. We identified 3 coactivator molecules, cAMP-response element binding protein (CBP), steroid receptor coactivator (SRC)-1, and peroxisome-proliferator activated receptor (PPAR)-gamma-coactivator (PGC)-1alpha, that bound in association with the PEPCK RAREs in vivo. Furthermore, there was differential binding of these coactivators in vitamin A-deficient mice. Related to this, specific lysine residues were acetylated on histones H3 and H4 at the 3 RAREs of the PEPCK promoter, consistent with the action of the above coactivators, and acetylation of certain lysines was significantly decreased with vitamin A deficiency. These results demonstrate the associated changes that occur in nuclear receptor binding, coactivator recruitment, and histone acetylation in response to vitamin A status, identified at specific RAREs in the PEPCK gene in vivo. PMID- 16317120 TI - The glycine N-methyltransferase (GNMT) 1289 C->T variant influences plasma total homocysteine concentrations in young women after restricting folate intake. AB - Glycine N-methyltransferase (GNMT) is a key regulatory protein in folate metabolism, methionine availability, and transmethylation reactions. Perturbations in GNMT may lead to aberrations in homocysteine metabolism, a marker of numerous pathologies. The primary objective of this study was to examine the influence of the GNMT 1289 C-->T alone, and in combination with the methylenetetrahydrofolate reductase (MTHFR) 677 C-->T variant, on plasma total homocysteine concentrations in healthy young women (n = 114). Plasma total homocysteine was measured at baseline (wk 0) and after 2 wk of controlled folate restriction (135 microg/d as dietary folate equivalents). Plasma homocysteine concentrations did not differ among the GNMT C1289T genotypes at baseline. However, after folate restriction, women with the GNMT 1289 TT genotype (n = 16) had higher (P = 0.019) homocysteine concentrations than women with the CT (n = 51) or CC (n = 47) genotype. The influence of the GNMT 1289 C-->T variant on homocysteine was dependent on the MTHFR C677T genotype. In subjects with the MTHFR 677 CC genotype, homocysteine was greater (P < or = 0.05) for GNMT 1289 TT subjects relative to 1289 CT or CC subjects. However, in subjects with the MTHFR 677 TT genotype, plasma homocysteine concentrations did not differ among the GNMT C1289T genotypes. Overall, these data suggest that the GNMT 1289 C-->T polymorphism influences plasma homocysteine and is responsive to folate intake. PMID- 16317121 TI - Isoflavones and functional foods alter the dominant intestinal microbiota in postmenopausal women. AB - Dietary phytoestrogens, such as isoflavones, are used as food additives to prevent menopause-related disorders. In addition to other factors, their bioavailability strongly depends on the activity of intestinal bacteria but the underlying interactions remain poorly understood. A randomized, double-blind, placebo-controlled study was undertaken with 39 postmenopausal women to characterize changes in the dominant microbial communities of the intestinal tract after 2 mo of isoflavone supplementation with and without pro- or prebiotic. The diversity and composition of the dominant microbiota were analyzed by temporal temperature-gradient gel electrophoresis (TTGE) and fluorescent in situ hybridization. Isoflavones alone stimulated dominant microorganisms of the Clostridium coccoides-Eubacterium rectale cluster, Lactobacillus-Enterococcus group, Faecalibacterium prausnitzii subgroup, and Bifidobacterium genus. The stimulation of the Clostridium coccoides-Eubacterium rectale cluster depended on the women's equol excretion and was transient, with the exception of a prolonged bifidogenic effect. Lasting changes in the diversity of the dominant species were also observed. The probiotic strain supplied could be detected by TTGE during its passage through the intestinal tract, and ingestion of fructooligosaccharides triggered a marked and specific bifidogenic effect. In conclusion, this is the first human study that shows changes in the diversity and composition of dominant bacterial communities in response to dietary supplementation with hormone-related compounds combined with functional foods. PMID- 16317122 TI - Maintenance of the LDL cholesterol:HDL cholesterol ratio in an elderly population given a dietary cholesterol challenge. AB - We previously evaluated the responses to dietary cholesterol in children and young adults. In this study, the effects of dietary cholesterol on plasma lipids and LDL atherogenicity were evaluated in 42 elderly subjects (29 postmenopausal women and 13 men > 60 y old). Our exclusion criteria were diabetes, heart disease, and the use of reductase inhibitors. The study followed a randomized crossover design in which subjects were assigned to consume the equivalent of 3 large eggs (EGG) daily or the same amount of a cholesterol-free, fat-free egg substitute (SUB) for a 1-mo period. After a 3-wk washout period, subjects were assigned to the alternate treatment. The concentration of plasma cholesterol after the EGG period varied among subjects. When all subjects were evaluated, there were significant increases in LDL cholesterol (LDL-C) (P < 0.05) and HDL-C (P < 0.001) for both men and women during the EGG period, resulting in no alterations in the LDL-C:HDL-C or the total cholesterol:HDL-C ratios. In addition, the LDL peak diameter was increased during the EGG period for all subjects. In contrast, the measured parameters of LDL oxidation, conjugated diene formation, and LDL lag time did not differ between the EGG and the SUB periods. We conclude from this study that dietary cholesterol provided by eggs does not increase the risk for heart disease in a healthy elderly population. PMID- 16317123 TI - The dietary alpha-linolenic acid to linoleic acid ratio does not affect the serum lipoprotein profile in humans. AB - Alpha-linolenic acid [ALA, 18:3(n-3)] and linoleic acid [LA, 18:2(n-6)] have comparable effects on serum lipid and lipoprotein concentrations, but their effects on lipoprotein subclass distributions and particle sizes are unknown. It is also not known whether these effects are changed by the ALA:LA ratio in the diet. To address these questions, healthy subjects (n = 54) consumed a control diet providing 7% of energy (En%) as LA and 0.4 En% as ALA during a 4-wk run-in period. For the following 6 wk of intervention, each diet was consumed by 18 subjects: the control diet, a low-LA diet (3 En% LA, 0.4 En% ALA), or a high-ALA diet (7 En% LA, 1.1 En% ALA). The ALA:LA ratio for the control diet was 1:19 and was 1:7 for the other 2 diets. Compared with the control group, LDL cholesterol decreased significantly in the ALA group (-0.32 mmol/L, P = 0.024), as did total cholesterol, apolipoprotein (apo) B, and the total:HDL cholesterol ratio. None of the dietary interventions affected HDL cholesterol, apo A-1, or triacylglycerol concentrations. The decrease in total VLDL particle concentrations in the low-LA group was due mainly to a decrease in medium VLDL (-16 nmol/L, P = 0.018) and in the high-ALA group to a decrease in small VLDL (-14 nmol/L, P = 0.044). We conclude that the ALA:LA ratio does not affect the serum lipoprotein profile. Compared with the control and LA diets, ALA lowered LDL cholesterol concentrations, possibly caused by the decrease in small VLDL. PMID- 16317124 TI - Stearic, oleic, and linoleic acids have comparable effects on markers of thrombotic tendency in healthy human subjects. AB - Because human studies concerning the effects of stearic acid on thrombotic tendency are inconsistent, we compared the effects of stearic acid with those of its unsaturated derivatives, oleic acid and linoleic acid. In this randomized, crossover study, 45 subjects (27 women and 18 men) consumed, in random order, 3 experimental diets, each for 5 wk. Diets contained approximately 38% of energy as fat. Dietary compositions were the same except for 7% of energy from stearic, oleic, or linoleic acids. At the end of each period, ex vivo and in vitro platelet aggregation, and variables of coagulation, fibrinolysis, and hematology were evaluated. In men, ex vivo platelet aggregation time as measured by filtragometry (P = 0.036 for diet effects) was favorably prolonged during consumption of the linoleic acid diet compared with the stearic acid diet (P = 0.040), but there was no difference with consumption of the oleic acid diet (P = 0.198). In vitro platelet aggregation induced by collagen and ADP, and variables of coagulation (factor VII amidolytic activity and concentrations of fibrinogen and prothrombin fragment 1 and 2) and fibrinolysis [plasminogen activator inhibitor (PAI) activity and concentrations of tissue plasminogen activator (tPA)/PAI-1 complexes] did not differ among the 3 diets. The mean platelet volume of the subjects decreased during consumption of the stearic acid diet by 0.32 fL compared with the oleic acid diet (P < 0.001) and by 0.35 fL compared with the linoleic acid diet (P < 0.001). In conclusion, our results do not suggest that stearic acid is highly thrombogenic compared with oleic and linoleic acids. PMID- 16317125 TI - The relative bioavailability of enterolignans in humans is enhanced by milling and crushing of flaxseed. AB - Flaxseed is one of the richest sources of lignans and is increasingly used in food products or as a supplement. Plant lignans can be converted by intestinal bacteria into the so-called enterolignans, enterodiol and enterolactone. For a proper evaluation of potential health effects of enterolignans, information on their bioavailability is essential. The aim of this study was to investigate whether crushing and milling of flaxseed enhances the bioavailability of enterolignans in plasma. In a randomized, crossover study, 12 healthy subjects supplemented their diet with 0.3 g whole, crushed, or ground flaxseed/(kg body weight . d). Each subject consumed flaxseed for 10 successive days separated by 11-d run-in/wash-out periods, in which the subjects consumed a diet poor in lignans. Blood samples were collected at the end of each run-in/wash-out period, and at the end of each supplement period. Plasma enterodiol and enterolactone were measured using LC-MS-MS. The mean relative bioavailability of enterolignans from whole compared with ground flaxseed was 28% (P < or = 0.01), whereas that of crushed compared with ground flaxseed was 43% (P < or = 0.01). Crushing and milling of flaxseed substantially improve the bioavailability of the enterolignans. PMID- 16317126 TI - The concentration of free holo-retinol binding protein is higher in vitamin A sufficient than in deficient Nepalese women in late pregnancy. AB - Free holo-retinol binding protein (RBP) [i.e., unbound to transthyretin (TTR)] plays a role in transporting vitamin A across the placenta during pregnancy. In a cross-sectional study of clinically healthy urban women, we assessed the association among clinical and biochemical factors on estimated concentrations of free holo-RBP during the last trimester of pregnancy. Serum samples obtained from a subsample of women (n = 259), who had participated in the Night Vision Threshold Test study in Nepal, were analyzed for determinations of retinol by HPLC, and RBP, TTR, and alpha-1 acid glycoprotein by radial immunodiffusion. Free holo-RBP concentrations were calculated using dissociation constants for free holo- and apo-RBP. Among these women, 30% were vitamin A deficient based on either the RBP:TTR index < or = 0.36 or serum retinol < 1.05 micromol/L. Using stepwise regression analyses, the RBP:TTR index explained 75% of the variance in free holo-RBP concentrations, whereas retinol explained only 14%. Women were classified as vitamin A sufficient (n = 185) or deficient (n = 74) using the RBP:TTR index and were stratified into 3 gestational groups (I: 24-28 wk, II: 29 33 wk, III: >33 wk). Concentrations of free holo-RBP were higher in vitamin A sufficient women than in vitamin A-deficient women (mean +/- SEM, 48.1 +/- 1.2 vs. 27.6 +/- 0.8 nmol/L; P < 0.001), and in a 3 x 2 factorial analysis, the interaction between gestational group and vitamin A status was significant. These results demonstrate that the RBP:TTR index is a useful proxy for free holo-RBP concentration and that vitamin A status affects its distribution. PMID- 16317127 TI - Iron-biofortified rice improves the iron stores of nonanemic Filipino women. AB - Iron deficiency is endemic in much of the world, and food system-based approaches to eradication may be viable with new plant breeding approaches to increase the micronutrient content in staple crops. It is thought that conventional plant breeding approaches provide varieties of rice that have 400-500% higher iron contents than varieties commonly consumed in much of Asia. The efficacy of consuming high-iron rice was tested during a 9-mo feeding trial with a double blind dietary intervention in 192 religious sisters living in 10 convents around metro Manila, the Philippines. Subjects were randomly assigned to consume either high-iron rice (3.21 mg/kg Fe) or a local variety of control rice (0.57 mg/kg Fe), and daily food consumption was monitored. The high-iron rice contributed 1.79 mg Fe/d to the diet in contrast to 0.37 mg Fe/d from the control rice. The 17% difference in total dietary iron consumption compared with controls (10.16 +/ 1.06 vs. 8.44 +/- 1.82 mg/d) resulted in a modest increase in serum ferritin (P = 0.10) and total body iron (P = 0.06) and no increase in hemoglobin (P = 0.59). However, the response was greater in nonanemic subjects for ferritin (P = 0.02) and body iron (P = 0.05), representing a 20% increase after controlling for baseline values and daily rice consumption. The greatest improvements in iron status were seen in those nonanemic women who had the lowest baseline iron status and in those who consumed the most iron from rice. Consumption of biofortified rice, without any other changes in diet, is efficacious in improving iron stores of women with iron-poor diets in the developing world. PMID- 16317128 TI - Food insecurity affects school children's academic performance, weight gain, and social skills. AB - Food insecurity has been associated with diverse developmental consequences for U.S. children primarily from cross-sectional studies. We used longitudinal data to investigate how food insecurity over time related to changes in reading and mathematics test performance, weight and BMI, and social skills in children. Data were from the Early Childhood Longitudinal Study-Kindergarten Cohort, a prospective sample of approximately 21,000 nationally representative children entering kindergarten in 1998 and followed through 3rd grade. Food insecurity was measured by parent interview using a modification of the USDA module in which households were classified as food insecure if they reported > or =1 affirmative response in the past year. Households were grouped into 4 categories based on the temporal occurrence of food insecurity in kindergarten and 3rd grade. Children's academic performance, height, and weight were assessed directly. Children's social skills were reported by teachers. Analyses examined the effects of modified food insecurity on changes in child outcomes using lagged, dynamic, and difference (i.e., fixed-effects) models and controlling for child and household contextual variables. In lagged models, food insecurity was predictive of poor developmental trajectories in children before controlling for other variables. Food insecurity thus serves as an important marker for identifying children who fare worse in terms of subsequent development. In all models with controls, food insecurity was associated with outcomes, and associations differed by gender. This study provides the strongest empirical evidence to date that food insecurity is linked to specific developmental consequences for children, and that these consequences may be both nutritional and nonnutritional. PMID- 16317129 TI - Growth indices, anemia, and diet independently predict motor milestone acquisition of infants in south central Nepal. AB - The acquisition of bipedal locomotion is an important aspect of gross motor development that ultimately affects the cognition of young children. Evidence for associations between nutrition-related variables and walking acquisition exist; however, questions remain about the importance of weight-for-length and dietary factors and the independent contribution of anemia and growth to walking. We examined the effect of nutritional factors on the acquisition of walking in a cross-sectional cohort of 4- to 17-mo old Nepali children (n = 485) adjusting for age, sex, caste, and socioeconomic status (SES). Participants were identified from census data collected in 1 village development committee in Sarlahi District and enrolled in a cross-sectional, community-based study between January and March 2002. Hemoglobin and erythrocyte protoporphyrin (EP) were measured at baseline using a heel-prick technique. The mean hemoglobin concentration was 101 +/- 12.5 g/L; 58% were anemic (hemoglobin < 105 g/L), 2.1% were severely anemic (hemoglobin < 70 g/L), and 43% of the children had iron-deficiency anemia (hemoglobin < 105 g/L; EP > or = 90 micromol/mol heme). Growth was delayed, i.e., 33.7% were stunted and 20.6% were wasted. Multivariate logistic models that controlled for age, sex, caste, and SES revealed that children with higher length for-age and weight-for-length Z-scores, no anemia, and meat consumption walked at an earlier age than children with lower scores, anemia, and no meat consumption. We conclude that growth, anemia, and diet are independently associated with delays in the onset of bipedal locomotion among young Nepali children. PMID- 16317130 TI - Colonic fermentation of inulin increases whole-body acetate turnover in dogs. AB - Metabolism of acetate from colonic fermentation was investigated in dogs. Beagle dogs (n = 9) were fed a control diet for 17 d followed by a 3% inulin-enriched diet (from chicory) for 4 and 21 d. On 3 occasions, the dogs were administered simultaneously infusions of [1-(13)C]acetate i.v. and [1,2-(13)C(2)]acetate intrarectally. Peripheral acetate concentration and turnover did not change over time after consumption of an inulin-enriched diet for 4 d. After 21 d of consuming the inulin-enriched diet, the whole-body acetate turnover increased significantly by 31% from (mean +/- SEM) 15.6 +/- 2.2 to 20.4 +/- 2.9 micromol/(kg . min) without a change in concentration. The rate of colonic acetate production that reached the peripheral circulation was 4.8 +/- 1.8 micromol/(kg . min). However, no [1,2-(13)C(2)]acetate tracer was recovered in the peripheral circulation. The fraction of oxidized tracer was higher in the gut (64 +/- 3%) than in peripheral circulation (46 +/- 3%) in dogs fed an inulin enriched diet for 21 d. In conclusion, colonic fermentation of inulin occurred and indirectly stimulated whole-body acetate turnover in dogs fed an inulin enriched diet for 21 d. PMID- 16317131 TI - Soy protein containing isoflavones reduces the size of atherosclerotic plaques without affecting coronary artery reactivity in adult male monkeys. AB - The cardiovascular effects of dietary soy on men or adult male experimental animals have received little attention. We determined the effects of long-term (31 mo) consumption of a commercially available soy protein concentrate containing experimentally varied concentrations of isoflavones on the development of atherosclerosis and vascular reactivity in adult male monkeys. The monkeys were fed atherogenic diets that differed only in the source of protein: Control (n = 30), casein and lactalbumin; low-isoflavone soy (n = 30), a mixture of unmodified soy protein isolate and isoflavone-depleted soy protein isolate containing 0.94 mg of isoflavones/g protein; and high-isoflavone soy (n = 31), unmodified soy protein isolate containing 1.88 mg of isoflavone/g protein. Plasma LDL cholesterol was reduced, whereas HDL cholesterol and apolipoprotein A-1 (P < 0.05) were increased in both groups that consumed soy protein. Atherosclerosis (mean plaque size in the coronary arteries) was reduced by approximately 34% (P < 0.05) in both groups fed soy protein. There were no effects of dietary soy on endothelium-dependent or -independent reactivity of coronary arteries. The results indicate that long-term consumption of soy protein containing a modest amount of isoflavones inhibits the early progression of coronary artery atherosclerosis without affecting endothelium-dependent or -independent arterial function. PMID- 16317132 TI - Isomalto-oligosaccharides polarize Th1-like responses in intestinal and systemic immunity in mice. AB - Isomalto-oligosaccharides (IMO) belong to a group of prebiotics that significantly increase the number of protective gut microflora. In the present study, we investigated the effects of IMO on intestinal and systemic immunity in mice. When mice were fed a diet supplemented with 20% IMO for 4 wk, the number of lactobacilli and the levels of IgA in feces were greater than those of mice fed the control diet (P < 0.05). Interferon-gamma (IFN-gamma) production by intestinal intraepithelial lymphocytes (i-IEL) in response to T-cell receptor (TCR) triggering was greater in mice fed IMO than in controls (P < 0.05), indicating T helper-1 (Th1) polarization of intestinal immunity by IMO. The proportion of natural killer (NK) T cells in the liver mononuclear cells (MNC), and the production of IFN-gamma by the liver MNC in response to TCR triggering were greater in mice fed IMO than in controls (P < 0.05), suggesting that the Th1/Th2 balance was shifted toward the Th1 lineage by IMO in systemic immunity. Furthermore, the proportion and activity of NK cells were greater in the spleens of the mice fed IMO than in the controls. Dietary IMO protected the mice from gamma-irradiation-induced lethality, accompanied by an inhibition of the translocation of Enterobacteriaceae. Notably, when mouse macrophage-like J774.1 cells were cultured with Lactobacillus gasseri in the presence of IMO, interleukin (IL)-12 production was greater than in the absence of IMO. These results suggest that IMO, in synergy with lactobacilli, upregulate the Th1 response and beneficially modulate host defense. PMID- 16317133 TI - Fat intake is associated with serum estrogen and androgen concentrations in postmenopausal Japanese women. AB - A reduction in fat intake has been associated with decreased estrogen levels in dietary intervention studies. However, previous cross-sectional studies conducted mainly among Western populations did not find a positive association between fat intake and postmenopausal estrogen levels. This study examined the cross sectional association of fat intake with serum levels of estrone, estradiol, testosterone, and dehydroepiandrosterone sulfate (DHEAS) in women. Study subjects were 324 healthy postmenopausal Japanese women. Diet including fat intake was assessed by a validated semiquantitative FFQ. After controlling for age and other potential breast cancer risk factors, serum estrone was positively associated with the percentage of energy from total fat (P = 0.04). The associations of serum estrone with monounsaturated fat and polyunsaturated fat were of borderline significance (P = 0.05). Serum DHEAS was positively associated with the percentage of energy from total fat (P = 0.007), saturated fat (P = 0.009), monounsaturated fat (P = 0.006), and polyunsaturated fat (P = 0.04). Serum estrone and DHEAS concentrations increased 11.8 and 9.3%, respectively, with a 5% increase in the percentage of energy from total fat. These data suggest that a high intake of fat is associated with higher serum levels of estrone and DHEAS in postmenopausal women. PMID- 16317134 TI - Development of the indicator amino acid oxidation technique to determine the availability of amino acids from dietary protein in pigs. AB - Standardized ileal ("true") digestibility is currently the best estimate of amino acid digestibility, but it does not measure bioavailability. Growth assays to determine amino acid bioavailability are expensive and laborious; thus, a rapid method is needed. Applying the principle of slope-ratio assay to the indicator amino acid oxidation (IAAO) method, we hypothesized that the reduction in indicator oxidation per gram of lysine in feedstuffs relative to that per gram of free lysine represented the bioavailability of lysine, here termed "metabolic availability." Indicator oxidation in pigs was linear over increasing lysine intakes (r = 0.90, P = 0.001) when the dietary lysine contents were 2 SD below the mean lysine requirement of the pigs. Peas were treated (raw, heated to reduce lysine availability, or heated with added lysine) to test the responsiveness of the IAAO to differing lysine availability. Free lysine reduced indicator oxidation by 3.16% of dose oxidized per gram added lysine, whereas the addition of protein lysine as raw (-2.81%) and heated peas (-1.73%) reduced oxidation to a lesser degree. Adding free lysine to heated peas decreased indicator oxidation, evidence that heating had worsened the utilization of pea protein for protein synthesis by reducing the bioavailability of lysine alone. Pea diets differed only in the availability of lysine; therefore IAAO detected differences in lysine bioavailability. Because the IAAO technique responds to lysine available at the sites of protein synthesis, the metabolic availability covers all losses during digestion, absorption, and utilization of lysine. This method can determine the metabolic availability of amino acids of a feedstuff within 2 wk. PMID- 16317135 TI - Orally administered green tea polyphenols prevent ultraviolet radiation-induced skin cancer in mice through activation of cytotoxic T cells and inhibition of angiogenesis in tumors. AB - Green tea polyphenols (GTPs) show promise as anticarcinogenic agents and may prevent the development of solar UV radiation-induced skin cancer. Here we investigated the mechanisms by which GTPs prevent UVB-induced skin cancer in mice. Two groups of 6- to 7-wk-old female SKH-1 hairless mice were UVB irradiated (180 mJ/cm(2)) 3 times each week for 24 wk. One group consumed water and the other, water containing 2 g/L GTPs. A control group drank water and was not exposed to UVB radiation. UVB-induced tumors and skin biopsies from the control group were analyzed using immunostaining, Western blotting, and gelatinolytic zymography. Oral administration of GTPs reduced UVB-induced tumor incidence (35%), tumor multiplicity (63%), and tumor growth (55%). The GTPs+UVB group had reduced expression of the matrix metalloproteinases (MMP)-2 and MMP-9, which have crucial roles in tumor growth and metastasis, and enhanced expression of tissue inhibitor of MMP in the tumors compared with mice that were treated with UVB alone. The GTPs+UVB group also had reduced expressions of CD31 and vascular endothelial growth factor, which are essential for angiogenesis, and inhibited expression of proliferating cell nuclear antigen in the tumors compared with the UVB group. Additionally, there were more cytotoxic CD8(+) T cells in the tumors of the GTPs+UVB group than in the UVB group and their tumor cells exhibited greater activation of caspase-3, indicating the apoptotic death of the tumor cells. Taken together, these data suggest that in mice, administration of GTPs affects several biomarkers that are involved in UV-carcinogenesis, including inhibition of angiogenic factors and recruitment of cytotoxic T cells in the tumor microenvironment. PMID- 16317136 TI - Oral administration of butyrivibrio fibrisolvens, a butyrate-producing bacterium, decreases the formation of aberrant crypt foci in the colon and rectum of mice. AB - Butyrivibrio fibrisolvens, a butyrate-producing ruminal bacterium, was evaluated for use as a probiotic to prevent colorectal cancer. Oral administration to Jcl:ICR mice of a new strain of B. fibrisolvens (MDT-1) that produces butyrate at a high rate (10(9) cfu/dose) increased the rate of butyrate production by fecal microbes, suggesting that MDT-1 can grow in the gut. The number of colorectal aberrant crypt foci (ACF), putative preneoplastic lesions induced by 1,2 dimethylhydrazine, was reduced after MDT-1 administration (10(9) cfu/dose, 3 times/wk for 4 wk). The number of aberrant crypts (ACs), number of foci having 3 or 4 ACs per focus, and the percentage of mice having 3 or 4 ACs per focus were also reduced, suggesting that the progress of lesions was suppressed by MDT-1. Interestingly, the MDT-1 cell homogenate did not have a similar beneficial effect. MDT-1 had low beta-glucuronidase activity, and administration of MDT-1 reduced the beta-glucuronidase activity in the colorectal contents. The numbers of natural killer (NK) and NKT cells in the spleen were markedly enhanced in response to MDT-1. Decreased beta-glucuronidase activity and increased numbers of NK and NKT cells and butyrate production may explain in part why MDT-1 administration suppressed ACF formation. These results suggest that colorectal cancer may be prevented or suppressed by the utilization of MDT-1 as a probiotic. Administration of MDT-1 had no harmful effect on the health of mice at least for 3 mo. PMID- 16317137 TI - Fisetin inhibits the activities of cyclin-dependent kinases leading to cell cycle arrest in HT-29 human colon cancer cells. AB - Fisetin, a natural flavonol present in edible vegetables, fruits, and wine, was reported to exert anticarcinogenic effects. The objective of the current study was to examine the effect of fisetin on the cell cycle progression of the human colon cancer cell line HT-29. HT-29 cells were cultured in serum-free medium with 0, 20, 40, or 60 micromol/L fisetin. Fisetin dose dependently inhibited both cell growth and DNA synthesis (P < 0.05), with a 79 +/- 1% decrease in cell number observed 72 h after the addition of 60 micromol/L fisetin. Perturbed cell cycle progression from the G(1) to S phase was observed at 8 h with 60 micromol/L fisetin treatment, whereas a G(2)/M phase arrest was observed after 24 h (P < 0.05). The phosphorylation state of the retinoblastoma proteins shifted from hyperphosphorylated to hypophosphorylated in cells treated with 40 micromol/L fisetin. (P < 0.05). Fisetin decreased the activities of cyclin-dependent kinases (CDK)2 and CDK4; these effects were likely attributable to decreases in the levels of cyclin E and D1 and an increase in p21(CIP1/WAF1) levels (P < 0.05). However, fisetin also inhibited CDk4 activity in a cell-free system (P < 0.05), indicating that it may directly inhibit CDk4 activity. The protein levels of cell division cycles (CDC)2 and CDC25C and the activity of CDC2 were also decreased in fisetin-treated cells (P < 0.05). These results indicate that inhibition of cell cycle progression in HT-29 cells after treatment with fisetin can be explained, at least in part, by modification of CDK activities. PMID- 16317139 TI - Workshop executive summary report. PMID- 16317150 TI - Opportunities for research. PMID- 16317151 TI - Review of the International Research Conference on Food, Nutrition, and Cancer, 2005. PMID- 16317152 TI - Integrating the ideas of life course across cellular, individual, and population levels in cancer causation. AB - Cells, individuals, and societies are complex systems in which the integrity of structure and function is protected through tight regulation and control. For each level of organization, health represents the ability to maintain integrity in response to the wider environment. Critical stages during growth and development act as checkpoints, where choice is exercised, and help determine future direction. Important among factors influencing the checkpoints include the availability of nutrients or foods within the immediate environment. At the cellular and whole-body levels, this information can be communicated to future generations. Recent work on the developmental origins of adult disease indicate specific factors that set limits on structure and function and potentially limit the capacity of the cell and individual to respond to environmental stressors that represent potential risk factors for neoplastic change. Epigenetic mechanisms modulate structure and function at the cellular and tissue levels, reflecting the potential for the growth and development of individuals, and reflect the food and nutrients available to the body as a whole and within the wider society. Understanding the nature and the interaction of the critical factors that determine and regulate variable stable and unstable gene expression will be increasingly important in characterizing abnormal cellular function and risk of disease for individuals and populations. This will require the ability to synthesize large data sets within and between different levels of organization to develop and refine a deeper understanding of how the systems are effectively integrated and regulated within and across generations and where this fails in the genesis of cancer. PMID- 16317153 TI - Mechanisms mediating the effects of prepubertal (n-3) polyunsaturated fatty acid diet on breast cancer risk in rats. AB - Dietary exposures during childhood may influence later breast cancer risk. We tested in an animal model the hypothesis that prepubertal intake of (n-3) PUFAs, present mainly in fish, reduces susceptibility to breast cancer. Between postnatal days 5 to 25, rat pups were fed (n-3) PUFA-containing diets at a 2:1 ratio of (n-6):(n-3) PUFAs (typical of prehistoric societies) or a control (n-6) PUFA diet at a 17:1 ratio of (n-6):(n-3) PUFAs (comparable with current Western societies). These fatty acids were given in a low- or high-fat context (16 or 39% energy from fat). The low-(n-3) PUFA diet reduced while the high-(n-3) PUFA diet increased carcinogen-induced mammary tumorigenesis. The low-(n-3) PUFA diet reduced mammary cell proliferation and increased apoptosis, particularly in the terminal end buds (the mammary source of malignant breast tumors). The high-(n-3) PUFA diet had opposite effects on these 2 key biomarkers and increased phospho Akt levels, a survival factor. Microarray analyses identified genes that were permanently upregulated in the low-(n-3) PUFA-exposed glands and function in oxidative damage repair. Serum levels of 8-hydroxy-2'deoxyguanosine, a marker of DNA damage, were significantly reduced in these low-(n-3) PUFA-fed rats, and increased in the high-(n-3) PUFA-exposed group. The latter group exhibited reduced expression of BRCA1, a DNA repair gene. Our results indicate that the opposing susceptibilities to mammary tumorigenesis between the low- versus high fat (n-3) PUFA-exposed groups were associated with altered DNA damage repair and gene expression linked to proliferation, survival, and differentiation. PMID- 16317154 TI - Chemoprevention of breast cancer, proteomic discovery of genistein action in the rat mammary gland. AB - Genistein, the primary isoflavone component of soy, consumed in the diet during the prepubertal period only, and the combined prepubertal and adult periods, suppresses chemically induced mammary cancer in rats. Gestational or adult-only exposures do not provide protection. An inverse relation exists between cancer susceptibility and mammary gland differentiation. The current study used proteomic technology to investigate genistein mechanisms of action as related to programming against chemically induced mammary cancer. Rats were injected subcutaneously with 500 microg genistein/g body weight on d 16, 18, and 20 postpartum. At d 21, mammary glands were subjected to 2-dimensional polyacrylamide gel electrophoresis. After gel scanning, image analysis, and MS, 6 proteins were determined to be differentially regulated and identified. One protein, GTP-cyclohydrolase 1 (GTP-CH1), was confirmed as being significantly upregulated at d 21 by immunoblot analysis. Investigation of downstream signaling from GTP-CH1 showed that tyrosine hydroxylase was upregulated and vascular endothelial growth factor receptor 2 (VEGFR2) was downregulated in the mammary glands of 50-d-old rats treated with genistein in the prepubertal period. This and previous work suggest that early prepubertal exposure to genistein enhances cell proliferation by upregulating GTP-CH1 and the epidermal growth factor (EGF) signaling pathway, and hence cell differentiation and gland maturation. This unique developmental maturation leads to a new biochemical blueprint, whereby the cells have reduced EGF signaling and VEGFR2, which renders the mature mammary gland less proliferative and less susceptible to cancer. This study demonstrated the usefulness of proteomics for the discovery of novel pathways that may be involved in cancer prevention. PMID- 16317155 TI - Interaction among folate, riboflavin, genotype, and cancer, with reference to colorectal and cervical cancer. AB - Epidemiological studies have linked low folate intake with an increased risk of epithelial cancers, including colorectal cancer and cervical cancer. Riboflavin has received much less attention, but there is increasing interest in the well established role that flavins play in folate metabolism and the possible synergy of a protective effect between these 2 vitamins. Folate plays a key role in DNA synthesis, repair, and methylation, and this forms the basis of mechanistic explanations for a putative role for folate in cancer prevention. The role of folate in these processes may be modulated by genotype for the common C677T thermolabile variant of methylene tetrahydrofolate reductase (MTHFR), homozygosity for which is associated with lower enzyme activity, lower plasma and red blood cell folate, and elevated plasma homocysteine. Riboflavin, as FAD, is a cofactor for MTHFR and there is evidently some interaction among riboflavin status, folate status, and genotype in determining plasma homocysteine, a functional marker of folate status. The MTHFR C677T polymorphism appears to interact with folate and riboflavin in modulating cancer risk in a manner that varies according to cancer site. Most evidence points to a protective effect of this polymorphism for risk of colorectal cancer, but the effect on cervical cancer risk is not clear. The effect of this polymorphism on cancer risk seems to be further modulated by other factors, including alcohol and, in the case of cervical cancer, infection with the human papilloma virus. An additional factor determining the effect of diet and genotype interactions on cancer risk may be the stage of cancer development. PMID- 16317156 TI - Genetic and epigenetic interactions between folate and aging in carcinogenesis. AB - Folate is among the most strongly implicated dietary components to convey protection against colon cancer, and diminished folate status is associated with an enhanced risk of colon cancer. Age is also regarded as one of the most important risk factors for colonic carcinogenesis. It is therefore of considerable interest to determine whether the process of aging influences folate metabolism in the colon and whether folate supplementation might prevent the procarcinogenic effects associated with aging. Recent studies in our laboratory demonstrated that the colonic mucosa of elder rats is more susceptible to folate depletion than that of young rats. Depletion of folate results in a shift in the forms of folate in the colon as well as increased uracil incorporation into DNA, a purported mechanism for colonic carcinogenesis. However, modest folate supplementation eliminates evidence of inadequate folate status in the colons of elder rats, suggesting that the relation between age and folate status in the colon might be one mechanism by which aging modulates colorectal cancer risk. Interactions between folate and aging also affect a spectrum of epigenetic and genetic phenomena such as uracil misincorporation, DNA methylation, protein methylation, mitochondrial deletion, and critical gene expression, which could be related to carcinogenesis. Aging and inadequate dietary folate may interact and collectively induce derangements in folate metabolism, thereby provoking subsequent molecular aberrations, which may enhance carcinogenesis. However, folate supplementation appears to reverse these adverse effects of aging, which is potentially of substantial import because the latter is an unmodifiable risk factor. PMID- 16317157 TI - Synergy among phytochemicals within crucifers: does it translate into chemoprotection? AB - The association between cruciferous vegetables and cancer prevention has been linked to glucosinolate derivatives. These phytochemicals enhance endogenous detoxification, leading to inactivation of potential carcinogens before initiation occurs. Two derivatives, indole-3-carbinol (I3C) and 1-cyano-2-hydroxy 3-butene (crambene) were shown in rats to induce a synergistic enhancement of detoxification enzyme activity. To follow up on these findings, a short-term carcinogenicity study using aflatoxin B1 (AFB(1)) was performed in which male F344 rats were fed diets supplemented with these 2 compounds alone or in combination. Groups included a negative control group (no AFB(1), crambene, or I3C), a crambene group (diet 0.150% crambene), an I3C group (diet 0.165% I3C), a high-dose group (diet 0.150% crambene, 0.165% I3C) a low-dose group (diet 0.030% crambene, 0.033% I3C), and a positive control group (AFB(1) treatment only). AFB(1) was administered after 2 wk of dietary pretreatment. Liver sections were scored for lesions including karyomegaly, apoptosis, and biliary hyperplasia and evaluated for expression of the preneoplastic marker glutathione S-transferase-pi (GSTP). I3C and crambene groups were protected against AFB(1) toxicity whereas the low-dose group was not. The high-dose group had scores close to those of the negative controls. For log(10) transformed 2- and 3-dimensional GSTP data, the high-dose group demonstrated synergistic reduction in GSTP-positive area and an additive reduction in GSTP-positive volume compared with the crambene and I3C groups. The low-dose group had no effect. In conclusion, high combination dietary doses of I3C and crambene demonstrated enhanced protection from AFB(1). Low combination doses, as might be realistically in the diet, were not effective. PMID- 16317158 TI - Green tea polyphenol epigallocatechin-3 gallate (EGCG) affects gene expression of breast cancer cells transformed by the carcinogen 7,12-dimethylbenz[a]anthracene. AB - Since the 1980s, the incidence of late-onset breast cancer has been increasing in the United States. Known risk factors, such as genetic modifications, have been estimated to account for approximately 5 to 10% of breast cancer cases, and these tend to be early onset. Thus, exposure to and bioaccumulation of ubiquitous environmental chemicals, such as polycyclic aromatic hydrocarbons (PAHs), have been proposed to play a role in this increased incidence. Treatment of female Sprague-Dawley rats with a single dose of the PAH 7,12-dimethylbenz[a]anthracene (DMBA) induces mammary tumors in approximately 90 to 95% of test animals. We showed previously that female rats treated with DMBA and given green tea as drinking fluid displayed significantly decreased mammary tumor burden and invasiveness and a significantly increased latency to first tumor. Here we used cDNA microarray analysis to elucidate the effects of the green tea polyphenol epigallocatechin-3 gallate (EGCG) on the gene expression profile in a DMBA transformed breast cancer cell line. RNA was isolated, in quadruplicate, from D3 1 cells treated with 60 mug/mL EGCG for 2, 7, or 24 h and subjected to analysis. Semiquantitative RT-PCR and Northern blot analyses confirmed the changes in the expression of 12 representative genes seen in the microarray experiments. Overall, our results documented EGCG-altered expression of genes involved in nuclear and cytoplasmic transport, transformation, redox signaling, response to hypoxia, and PAHs. PMID- 16317159 TI - Zingiberaceous and citrus constituents, 1'-acetoxychavicol acetate, zerumbone, auraptene, and nobiletin, suppress lipopolysaccharide-induced cyclooxygenase-2 expression in RAW264.7 murine macrophages through different modes of action. AB - In the present study, we explored the suppressive activities of 1' acetoxychavicol acetate (ACA), auraptene, nobiletin, and zerumbone toward LPS induced cyclooxygenase (COX)-2 mRNA expression in mouse macrophages and the underlying molecular mechanisms. Pretreatment of RAW264.7 cells with LPS led to the activation of mitogen-activated protein kinase (MAPK)s [p38, extracellular signal-regulated kinase (ERK)1/2, c-Jun NH2-terminal kinase (JNK)1/2] and Akt, together with degradation of the inhibitor of nuclear factor-kappaB (IkappaB) alpha protein and nuclear translocation of nuclear factor (NF)-kappaB p65, and the resultant activation of activator protein (AP)-1, NF-kappaB, and cAMP responsive element-binding protein (CREB) transcription factors. ACA abrogated ERK1/2 and JNK1/2, but not p38 MAPK, as well as the activation of those transcription factors. Although it allowed LPS-triggered phosphorylation of those MAPKs and NF-kappaB nuclear translocation, nobiletin suppressed the activation of AP-1, NF-kappaB, and CREB. Zerumbone had no effect on those transcription factors, though it attenuated COX-2 mRNA expression, suggesting that it disrupts the stabilization of COX-2 mRNA. Conversely, zerumbone significantly accelerated spontaneous COX-2 mRNA decay, the potency of which was comparable with that of SB203580, an inhibitor of p38 MAPK, whose activation has key roles in the proinflammatory mRNA stabilization processes. Because SB203580 but not zerumbone suppressed LPS-induced p38 MAPK activation, the molecular targets of zerumbone may be MAPK-activated protein kinase-2 or located downstream. However, auraptene suppressed the expression of COX-2 protein but not mRNA, implying that it targets translation. We propose that these phytochemicals are promising chemopreventive agents for inflammation-associated carcinogenesis. Their use in combination may enhance their efficacy because of their different modes of action. PMID- 16317160 TI - Redox-sensitive transcription factors as prime targets for chemoprevention with anti-inflammatory and antioxidative phytochemicals. AB - Oxidative stress has been implicated in various pathological conditions including cancer. However, the human body has an intrinsic ability to fight against oxidative stress. A wide array of phase 2 detoxifying or antioxidant enzymes constitutes a fundamental cellular defense system against oxidative and electrophilic insults. Transcriptional activation of genes encoding detoxifying and antioxidant enzymes by NF-E2 related factor 2 (Nrf2), a member of the cap'n'collar family of basic leucine zipper transcription factors, may protect cells and tissues from oxidative damage. Many chemopreventive and chemoprotective phytochemicals have been found to enhance cellular antioxidant capacity through activation of this particular transcription factor, thereby blocking initiation of carcinogenesis. A new horizon in chemoprevention research is the recent discovery of molecular links between inflammation and cancer. Components of the cell signaling pathways, especially those that converge on redox-sensitive transcription factors, including nuclear factor-kappaB (NF-kappaB) and activator protein 1 (AP-1) involved in mediating inflammatory response, have been implicated in carcinogenesis. A wide variety of chemopreventive and chemoprotective agents can alter or correct undesired cellular functions caused by abnormal proinflammatory signal transmission mediated by inappropriately activated NF-kappaB and AP-1. The modulation of cellular signaling by anti inflammatory phytochemicals hence provides a rational and pragmatic strategy for molecular target-based chemoprevention. PMID- 16317161 TI - Impact of physical activity on intestinal cancer development in mice. AB - Observational epidemiology supports the hypothesis that variation in diet and other lifestyle exposures accounts for a large part of the variation in incidence of colorectal cancer (CRC). Physical inactivity is associated strongly with enhanced CRC risk, but no human intervention studies have shown causality. This paper reviews data from all available studies of the effects of exercise interventions on intestinal neoplasia using rat and mouse models. All 5 published studies of effects of increased physical activity (both forced and voluntary) using carcinogen-treated rat models show strong protection against CRC by greater physical activity. In contrast, there is little convincing evidence of reduced intestinal neoplasia after increased physical activity in the 3 published studies using Apc(Min) mice (which develop multiple intestinal polyps spontaneously) although the nature and amounts of physical activity imposed in rats and mice were similar. Major differences in protocol between the 2 groups of studies are that the rat studies were much longer (at least 20 wk and in most cases 38 wk compared with < or =9 wk for the mouse studies) and the primary endpoint was colorectal carcinoma (rats) rather than small bowel adenomas (mice). The epidemiological evidence for protection against adenoma formation by increased physical activity is weaker than that for carcinoma. The limited evidence available suggests that, compared with rats, mice may show a greater compensation for energy expenditure in exercise through reduction in nonexercise physical activity, thus ameliorating effects. The resulting smaller effects on body weight and body fatness may limit changes in intestinal neoplasia in Apc(Min) mice. PMID- 16317162 TI - Challenges in prostate cancer research: animal models for nutritional studies of chemoprevention and disease progression. AB - Prostate cancer is the second leading cause of cancer-related death in the United States. The American Cancer Society estimates that there will be over 232,000 new cases of prostate cancer in 2005. Evidence suggests that diet can act as a chemopreventive agent to reduce the incidence of prostate cancer as well as to reduce the mortality of the disease. Epidemiologic studies suggest that diets rich in specific vitamins, grains, fruits, and vegetables may be associated with lower cancer rates than high-fat diets, yet the molecular bases for these positive nutritional actions are largely unknown. The interactions of diet in combination with genetic determinants of disease progression are unclear because prostate cancer is also a disease resulting from abnormal gene expression. Hence, the biology of normal prostate development and the mechanisms underlying the initiation, progression, and metastatic spread of prostate cancer must be understood at the molecular level to develop effective nutritional prevention and intervention strategies to control and treat this malignant disease. However, progress toward understanding the biology of prostate cancer and the development of new therapies has been hampered by the lack of in vivo model systems that adequately capitulate the spectrum of benign, latent, aggressive, and metastatic forms of the human disease. In this review we discuss the diverse animal models of prostate cancer available and their applicability for nutritional studies of cancer prevention. PMID- 16317163 TI - Nutrient-gene interaction: metabolic genotype-phenotype relationship. AB - The U.S. Department of Health and Human Services (DHHS)/USDA Dietary Guidelines for Americans is a science and population evidence-based guide on diet and physical activity, providing advice and recommendations to promote a healthier lifestyle and reduce the risk of chronic diseases, including cancer. These recommendations are supported by the comprehensive evidence-based review on diet and cancer prevention conducted by the American Institute for Cancer Research, National Cancer Institute, World Health Organization/International Agency for Research on Cancer, and others. However, influencing dietary effects are the individual genetic predispositions that are the basis for considerable interindividual variations in cancer risk within the population and in nutrient homeostasis, which is maintained by genomic-nutrient and metabolic-phenotype interactions. Although genetics is an important component, it accounts for only a portion of this variation. An individual's overall phenotype, including health status, is achieved and maintained by the sum of metabolic activities functioning under differing circumstances within the life cycle and the complex interactions among genotype, metabolic phenotype, and the environment. In this postgenomic era, high-throughput groups of technologies in genomics, proteomics, and metabolomics measure and analyze DNA sequences, RNA transcripts, proteins, and nutrient-metabolic fluxes in a single experiment. These advances have transformed biomarker studies on nutrient-gene interactions from a reductionist concept into a holistic practice in which many regulated genes involved in metabolism, along with its metabolic phenotypes, can be measured through functional genomics and metabolic profiling. The overall integration of data and information from the building blocks of metabolism-based nutrient-gene interaction can lead to future individualized dietary recommendations to diminish cancer risk. PMID- 16317164 TI - Allelic loss of the gene for the GPX1 selenium-containing protein is a common event in cancer. AB - Selenium has been shown to reduce cancer incidence in animal models and more recent data indicate that it may be protective in humans as well. However, little is known about the mechanism by which selenium prevents cancer. Cytosolic glutathione peroxidase (GPX1), a selenium-containing antioxidant enzyme, has been implicated in the development of cancer of the head and neck, lung, and breast, in part because of allelic loss at the GPX1 locus. The study of allelic loss at the GPX1 locus in colon cancer was investigated by examining loss of heterozygosity (LOH) in DNA extracted from both tumor and adjacent histopathologically normal tissue obtained by laser capture microdissection. Tissue samples were obtained from 53 colon cancer patients. Two highly polymorphic markers, alanine codon repeats and a proline-leucine polymorphism (198P/L) present in the GPX1 gene, were used to examine LOH at this locus. Analysis of both polymorphisms identified LOH at GPX1 in a significant percentage of colorectal cancer (42%). These results indicated that LOH at the GPX1 locus is a common event in cancer development and that GPX1 or other tightly linked genes may be involved in the etiology of this disease. PMID- 16317165 TI - Pathways in nutritional modulation of homeostasis and tumorigenesis. PMID- 16317166 TI - Nutrient-gene interaction: tracer-based metabolomics. AB - Understanding nutrient-gene interaction requires tools for both the study of nutrigenomics and the characterization of phenotype. Metabolomics or metabolite profiling is a powerful tool for characterizing metabolic phenotype, and tracer based metabolomics is a subset of metabolomics that focuses on metabolite distribution and flux determination using tracers. In this review, the characterizations of metabolic phenotype by metabolite profiling and by metabolic flux measurements are compared. The rationale and methodologies of tracer-based metabolomics are explained. Tracer-based metabolomics provides a relational database of metabolites linked by the relationship of shared metabolic pathways, common substrates, and cofactors. Such a collection of flux measurements provides precise and accurate information on the operation of the cellular metabolic network and its response to genetic and nutrient environment changes. Nutrient gene interaction can be studied using the concept of constraint-based modeling, which states that the observed metabolic phenotype is a consequence of constraints from genetic factors and the nutrient environment. Thus, genetic inheritance (genomic constraints) confers a wide range of possible phenotypes whereas selection by metabolic (structural and pathway relationship) and environmental (physical environment and nutrient availability) constraints determines the final observed phenotype. The study of the contribution from nutrient and genetic factors to the survival advantage of cancer cells using flux measurements is a critical first step in our understanding of the relationship between nutrient intake and cancer risk. PMID- 16317167 TI - International Conference on Diet, Nutrition, and Cancer, July 14-15, 2005, Washington, DC, USA. Proceedings and abstracts. PMID- 16317171 TI - Contribution of regulatory and structural variations in APOE to predicting dyslipidemia. AB - The objective of this study was to evaluate 1) whether non single nucleotide polymorphisms-coding (non-cSNP) in the apolipoprotein E gene (APOE) identified by resequencing studies contribute to statistically explaining dyslipidemia if variations in the two cSNPs in exon 4 that define the 2, 3, and 4 alleles are ignored, and 2) whether the contribution of these additional SNPs persists when variations in the cSNPs are considered. We used an ecological, multiple population, data-mining strategy to identify single-SNP and two-SNP genotypes that distinguish between high and low levels of plasma lipids in three training samples, European-Americans from Rochester, MN, African-Americans from Jackson, MS, and Europeans from North Karelia, Finland. We found that a pair of SNPs located in the 5' region define genotypes A560T832/A560T832, A560T832/A560G832, and A560T832/T560T832, which distinguish between high and low levels of HDL cholesterol (HDL-C), triglycerides (TG), and/or total cholesterol (T-C). The A560T832/- genotypes predicted high TG and high T-C in both genders in a large independent test sample from Copenhagen, Denmark. Prediction of high T-C in the Danish females was dependent on genotypes defined by the cSNPs. Our study suggests that both regulatory and structural variations should be considered when evaluating the utility of APOE for predicting dyslipidemia in the population at large. PMID- 16317173 TI - The politics of developing research methods. PMID- 16317172 TI - Identification of cholesteryl esters in human carotid atherosclerosis by ex vivo image-guided proton MRS. AB - Vulnerable atherosclerotic plaques may be identified by their large lipid component, particularly liquid cholesteryl ester (CE), covered by a fibrous cap. We hypothesized that image-guided 1H proton magnetic resonance spectroscopy (MRS) would identify mobile CE in discrete, preselected regions of atherosclerotic plaque. Human carotid endarterectomy specimens (n = 10) were imaged ex vivo by magnetic resonance imaging (MRI) at high field (11.7 T) utilizing standard T1- and T2-weighted spin echo protocols. MRS spectra were acquired from 1 mm3 voxels, localized to plaque regions that we judged by MRI to be lipid rich or lipid poor. The spectra revealed methyl and methylene resonances of fatty acyl chains with relative intensities and linewidths characteristic of pure CE, by comparison with lipid standards. Regions judged to be lipid rich by MRI showed much more intense CE resonances than did lipid-poor regions. The integrated intensities of lipid peaks were 5.5 +/- 2.0% (lipid-rich regions) versus 0.9 +/- 0.6% (lipid-poor regions) of the unsuppressed water peak (P < 0.0001). Lipid distribution by histology, MRS, and MRI showed strong correlation. Image-guided proton MRS accurately identified CE in selected regions of atherosclerotic plaque as small as 1 mm3 in an ex vivo setting. This procedure may permit the noninvasive detection and quantification of CE in atherosclerotic plaque in vivo. PMID- 16317174 TI - The inextricable link in caring for families of critically burned children. AB - Advances in burn care have made recovery possible for children with burns on up to 99% of their body surface, which has prompted investigation of the psychosocial well-being of survivors. Although therapies have been suggested, no researchers have acknowledged or recognized nurses' potential contributions to children's and families' recovery. In this article, the author examines the lived experiences of 16 pediatric nurses on a burn intensive care unit. The purpose of the study was to articulate practices that helped critically burned children to heal holistically. Data showed that while helping burned children to heal holistically, the nurses simultaneously supported families to heal holistically. Nurses were observed laughing, talking, and crying with families, as well as role modeling ways of being with and caring for the children. These practices illustrated the nurses' understanding that care of the children was inextricably linked to care of the families. PMID- 16317175 TI - The construction of troubled and credible patients: a study of emotion talk in palliative care settings. AB - In this article, the authors select two categories of dying patients, "troubled" and "credible," from two larger studies conducted in three palliative care settings. They explore how nurses construct dying patients' moral identities and how they use emotion talk to interpret patients' behavior. The authors carried out a microanalysis of talk-in-action using discourse analysis and conversation analysis. Strategies used for the construction of moral identities include the production of atrocity stories and emotional editing. The authors identify moments when emotions are made relevant in palliative care nurses' daily practices, which serve to smooth social interaction and give a voice to dying patients' words and actions. The dying trajectory, the deteriorating emotional body, and the sound mind are resources used in the characterization of the credible and troubled patient. The authors argue that emotion talk is significant because it reveals how nurses manage conflict and tension in talk-in-interaction. PMID- 16317176 TI - When family-centered care is challenged by infectious disease: pediatric health care delivery during the SARS outbreaks. AB - In this ethnographic study, the authors examined the experiences and perspectives of children hospitalized because of SARS (severe acute respiratory syndrome), their parents, and pediatric health care providers. The sample included 5 children, 10 parents, and 8 health care providers who were directly affected by SARS during the time of the outbreaks and extreme infection control procedures. The data analyses illuminated a range of perceived experiences for this triadic sample. Issues related to social isolation due to infection control precautions were predominant. Themes included emotional upheaval, communication challenges, and changes in parental and professional roles. These findings reveal the cogent effects of SARS on family-centered care. The notion of providing family-centered care within an environment plagued by an infectious outbreak suggests an omniously difficult task. Efforts must be made to optimize family-centered care despite obstacles. The authors suggest effective clinical approaches in the event of future outbreaks. PMID- 16317177 TI - Images of cloning and stem cell research in editorial cartoons in the United States. AB - Through semiotic analysis of manifest and latent meanings in editorial cartoons, the author uncovers how cloning and stem cell research are represented in a popular mass medium. She identified 86 editorial cartoons published in the United States between 2001 and 2004 that referred to cloning and 20 that referred to stem cell research. Cartoonists portrayed people individually 224 times and 4 times in groups of more than 10. Men were portrayed in 64% of cartoons. Stem cell research was depicted as having a potential positive value, and cloning was depicted negatively. Some major messages are that cloning will lead to the mass production of evil, cloning creates monsters, and politics will influence who or what will be cloned. Analyzing popular images can allow access to public understanding about genetic technology and evaluation of public beliefs, preconceptions, and expectations as the public is educated on the use and value of services. PMID- 16317178 TI - Striving for emotional survival in palliative cancer nursing. AB - In this grounded theory study, the authors analyze interviews and participant observation data related to palliative cancer nursing in hospitals. Striving for Emotional Survival emerged as the pattern of behavior through which nurses deal with their main concern, the risk of being emotionally overloaded by their work. It involved three main strategies: Emotional Shielding through Professional Shielding or Cold Shielding; Emotional Processing through Chatting, Confirmation Seeking, Self-Reflecting, or Ruminating; and Emotional Postponing through Storing or Stashing. Emotional Competence is a property of Striving for Emotional Survival that explains more or less adequate ways of dealing with emotional overload. The theory Striving for Emotional Survival can be useful in the nurses' daily work and provides a comprehensive framework for understanding how nurses deal with emotional difficulties. The authors suggest that health care organizations encourage self-care, prioritize time to talk, and offer counseling to nursing staff with emotionally difficult working conditions. PMID- 16317179 TI - Genetic testing for hereditary breast and ovarian cancer: responsibility and choice. AB - Genetic testing for hereditary breast-ovarian cancer has become an important part of clinical genetics practice. Although considerable work has focused on the psychological impact of this technology, there has been little research into the moral implications of genetic information on hereditary cancer families. In this article, the author examines moral issues related to individuals' decisions to seek or decline testing. In-depth interviews with 53 participants make up the core of the research. Analysis of participants' accounts illustrates how the decision to be tested (or not) interconnects with moral agency and aspects of self (embodied, familial-relational, and civic self). The findings form the foundation for inquiry into conceptualization of moral responsibility, autonomy, and choice. They also provide insight that might assist clinicians to understand more fully the needs and responses of those who seek genetic testing for hereditary breast-ovarian cancer. PMID- 16317180 TI - Waiting for a liver transplant. AB - The waiting period for liver transplantation is a difficult time fraught with uncertainty and associated with a high rate of morbidity and mortality. To understand better what it means for a transplant patient to wait, the authors explore in this phenomenological study the meaning that people with liver failure ascribe to the experience of waiting for a transplant. They conducted 9 interviews using phenomenological methods of inquiry as a guide for analysis. Eight core themes emerged from 146 significant statements and corresponding meaning units. The experience of waiting includes transformations, doctors, teams and trust, elation to despair, loss, questioning the process, searching, coping, and the paradox of time. The essence of the experience is discussed in light of the theory of chronic illness as a disruption of biographical narrative. The authors highlight implications for the transplant teams and other health care providers. PMID- 16317181 TI - Clinical application research: a hermeneutical approach to the appropriation of caring science. AB - Clinical caring science researchers contribute, by means of various participatory research efforts, to bring clinical practice closer to the ideals of caring. These research efforts have in the main been developed from classical action research rooted in critical theory. In this article, the authors launch an alternative research approach called clinical application research, the basis of which can be traced to the interpretative paradigm, or hermeneutics. The basic cornerstones of this research approach are ontology, context, and appropriation as well as understanding, interpretation, and application. Using an example from ongoing clinical research, the authors demonstrate the utility of this approach. Their aim in this article is to contribute to the development of methods within clinical research. PMID- 16317182 TI - Carspecken's five-stage critical qualitative research method: an application to nursing research. AB - In this article, the authors provide an account of Carspecken's (1996) five-stage approach to "doing" critical ethnography, or what he has termed critical qualitative research (CQR). They provide the reader with an overview of the concepts presented in Carspecken's book Critical Ethnography in Educational Research and describe how they applied several of his ideas within a research project that explored renal nurses' decision making using a critical ethnographic approach. They briefly describe the five stages of CQR and incorporate within the article an example of how they applied the stages. They propose this approach as a useful method for nursing and other health-related research. PMID- 16317183 TI - Developing optimal search strategies for retrieving clinically relevant qualitative studies in EMBASE. AB - Qualitative researchers address many issues relevant to patient health care. Their studies appear in an array of journals, making literature searching difficult. Large databases such as EMBASE provide a means of retrieving qualitative research, but these studies represent only a minuscule fraction of published articles, making electronic retrieval problematic. Little work has been done on developing search strategies for the detection of qualitative studies. The objective of this study was to develop optimal search strategies to retrieve qualitative studies in EMBASE for the 2000 publishing year. The authors conducted an analytic survey, comparing hand searches of journals with retrievals from EMBASE for candidate search terms and combinations. Search strategies reached peak sensitivities at 94.2% and peak specificities of 99.7%. Combining search terms to optimize the combination of sensitivity and specificity resulted in values over 89% for both. The authors identified search strategies with high performance for retrieving qualitative studies in EMBASE. PMID- 16317184 TI - The influence of mood on the search for supporting versus conflicting information: dissonance reduction as a means of mood regulation? AB - Focusing on similarities between the mood regulation approach and dissonance theory, this article addresses the interplay between dissonance and mood by examining how individuals search for information after making a decision while under the influence of positive versus negative mood. Study 1 suggested that negative mood increased the preference for consonant over dissonant information after decisions, whereas positive mood led to a more balanced information search. In Study 2, participants in negative mood rated consonant information as more pleasant and dissonant information as more annoying than participants in positive mood. In addition, the results suggested that mood regulation processes took place. In Study 3, the findings from Study 1 were replicated with a paradigm in which higher stakes were involved. PMID- 16317185 TI - An ingroup advantage for confidence in emotion recognition judgments: the moderating effect of familiarity with the expressions of outgroup members. AB - The confidence we have in our assessment of an interaction partner's emotional state can have important consequences for the quality of the interaction. Two studies assessed the hypothesis that immigrants are more confident in their judgment of others' emotional facial expressions if the expresser is a member of their cultural ingroup rather than a member of the host community or another cultural group. In addition, the effects of the perceived familiarity with the type of expression, the length of residence in the host country, the quality of cross-cultural contact, the level of acculturation, and the intensity of the facial expressions were assessed. Overall, the results revealed an ingroup advantage effect for confidence ratings as well as support for the notion that individuals are more confident when judging expressions that they consider as more frequently displayed in everyday life. Furthermore, individuals were more confident when judging happiness expressions as well as more intense expressions in general. PMID- 16317186 TI - Different selves have different effects: self-activation and defensive social comparison. AB - Three studies show that different forms of self-activation have differential influences on the processing of social comparison information. Activating neutral self-conceptions results in defensive processing of threatening social comparison information (Study 1). Participants maintain favorable self-evaluations in the face of upward comparison and rate the upward target of comparison negatively. Activating positive self-conceptions results in non-defensive processing of threatening social comparison information (Study 2). Participants endorse negative self-evaluations following upward comparison and rate the upward target of comparison positively. Activating negative self-conceptions maximizes defensive processing of threatening social comparison information (Study 3). Participants maintain favorable self-evaluations in the face of upward comparison and rate both upward and downward targets of comparison negatively. These results are discussed in terms of their implications for strategies to maintain self esteem in the face of threatening comparisons. PMID- 16317187 TI - Marginalized relationships: the impact of social disapproval on romantic relationship commitment. AB - Little research has examined the effects of prejudice and discrimination on people's romantic relationships. The authors explored whether belonging to a socially devalued relationship affects consequential relational phenomena. Within the framework of the Investment Model, the authors (a) tested the association between perceived relationship marginalization and relationship commitment, (b) compared investment levels of individuals involved in marginalized versus nonmarginalized relationships, and (c) explored ways in which couples may compensate for decreased investments to maintain high commitment. Consistent with hypotheses, marginalization was a significant negative predictor of commitment. Moreover, individuals in marginalized relationships invested significantly less than individuals in nonmarginalized relationships. Despite investing less, marginalized relationship partners were significantly more committed than were their nonmarginalized counterparts. Thus, marginalized partners appeared to compensate for their reduced investments, with evidence suggesting that compensation occurs via reduced perception of relationship alternatives rather than via increased perception of relationship satisfaction. PMID- 16317188 TI - Why is enacted social support associated with increased distress? Using simulation to test two possible sources of spuriousness. AB - Although perceiving that social support is available is often associated with psychological benefits in times of stress, there is evidence that actually receiving support is associated with increased distress. To assess whether this latter association could be spurious, the authors conducted computer simulation studies to examine two theoretical models that could give rise to it. Study 1 examined a process whereby distress leads to provision of support rather than the reverse. Study 2 examined a process whereby an adverse event increases both support and distress. In both cases, substantial spurious associations were produced only when parameter values were markedly unrealistic. It was concluded that these possible sources of spuriousness are insufficient to explain the association between support and distress found in naturalistic studies. PMID- 16317189 TI - "Keep your eyes on the prize": reference points and racial differences in assessing progress toward equality. AB - White Americans tend to perceive greater progress toward racial equality than do ethnic minorities. Correlational evidence (Study 1) and two experimental manipulations of framing (Studies 2 and 3) supported the hypothesis that this perception gap is associated with different reference points the two groups spontaneously use to assess progress, with Whites anchoring on comparisons with the past and ethnic minorities anchoring on ideal standards. Consistent with the hypothesis that the groups anchor on different reference points, the gap in perceptions of progress was affected by the time participants spent deliberating about the topic (Study 4). Implications for survey methods and political conflict are discussed. PMID- 16317190 TI - Cultural estrangement: the role of personal and societal value discrepancies. AB - Study 1 examined whether cultural estrangement arises from discrepancies between personal and societal values (e.g., freedom) rather than from discrepancies in attitudes toward political (e.g., censorship) or mundane (e.g., pizza) objects. The relations between different types of value discrepancies, estrangement, subjective well-being, and need for uniqueness also were examined. Results indicated that personal-societal discrepancies in values and political attitudes predicted estrangement, whereas mundane attitude discrepancies were not related to estrangement. As expected, value discrepancies were the most powerful predictor of estrangement. Value discrepancies were not related to subjective well-being but fulfilled a need for uniqueness. Study 2 replicated the relations between value discrepancies, subjective well-being, and need for uniqueness while showing that a self-report measure of participants' values and a peer-report measure of the participants' values yielded the same pattern of value discrepancies. Together, the studies reveal theoretical and empirical benefits of conceptualizing cultural estrangement in terms of value discrepancies. PMID- 16317191 TI - Tempting today, troubling tomorrow: the roots of the precarious couple effect. AB - The precarious couple effect occurs when men pair with women who are both critical and more verbally disinhibited than them. Evidence that dissatisfaction runs high in such relationships makes one ask why people enter them in the first place. In Study 1, respondents recalled that past partners who were verbally disinhibited were relatively active in initiating the relationship. In Study 2, verbally inhibited men evidenced ambivalence in that they disliked disinhibited women more than inhibited ones but these feelings of disliking did not translate into expectations of feeling tense during the interaction. Study 3 revealed that initial interactions between inhibited men and disinhibited women go smoothly unless (a) the women are critical and (b) the pair discusses a stressful topic. The authors suggest that members of precarious couples are drawn to one another because, in initial encounters, their communication styles are relatively symbiotic. Alas, this initial chemistry does not always endure. PMID- 16317192 TI - Narcissism and childhood recollections: a quantitative test of psychoanalytic predictions. AB - Different psychotherapeutic theories provide contradictory accounts of adult narcissism as the product of either parental coldness or excessive parental admiration during childhood. Yet, none of these theories has been tested systematically in a nonclinical sample. The authors compared four structural equation models predicting overt and covert narcissism among 120 United Kingdom adults. Both forms of narcissism were predicted by both recollections of parental coldness and recollections of excessive parental admiration. Moreover, a suppression relationship was detected between these predictors: The effects of each were stronger when modeled together than separately. These effects were found after controlling for working models of attachment; covert narcissism was predicted also by attachment anxiety. This combination of childhood experiences may help to explain the paradoxical combination of grandiosity and fragility in adult narcissism. PMID- 16317193 TI - A multilevel approach to the relationship between birth order and intelligence. AB - Many studies show relationships between birth order and intelligence but use cross-sectional designs or manifest other threats to internal validity. Multilevel analyses with a control variable show that when these threats are removed, two major results emerge: (a) birth order has no significant influence on children's intelligence and (b) earlier reported birth order effects on intelligence are attributable to factors that vary between, not within, families. Analyses on 7- to 8 - and 13- to 14-year-old children from the National Longitudinal Survey of Youth support these conclusions. When hierarchical data structures, age variance of children, and within-family versus between-family variance sources are taken into account, previous research is seen in a new light. PMID- 16317194 TI - Usage of "MSM" and "WSW" and the broader context of public health research. PMID- 16317195 TI - US Department of Health and Human Services: a need for global health leadership in preparedness and health diplomacy. PMID- 16317196 TI - Homicide and Native Americans. PMID- 16317197 TI - Minimum purchasing age for alcohol and traffic crash injuries among 15- to 19 year-olds in New Zealand. AB - OBJECTIVES: In 1999, New Zealand lowered the minimum purchasing age for alcohol from 20 to 18 years. We tested the hypothesis that this increased traffic crash injuries among 15- to 19-year-olds. METHODS: Poisson regression was used to compute incidence rate ratios for the after to before incidence of alcohol involved crashes and hospitalized injuries among 18- to 19-year-olds and 15- to 17-year-olds (20- to 24-year-olds were the reference). RESULTS: Among young men, the ratio of the alcohol-involved crash rate after the law change to the period before was 12% larger (95% confidence interval [CI]=1.00, 1.25) for 18- to 19 year-olds and 14% larger (95% CI=1.01, 1.30) for 15- to 17-year-olds, relative to 20- to 24-year-olds. Among young women, the equivalent ratios were 51% larger (95% CI=1.17, 1.94) for 18- to 19-year-olds and 24% larger (95% CI=0.96, 1.59) for 15- to 17-year-olds. A similar pattern was observed for hospitalized injuries. CONCLUSIONS: Significantly more alcohol-involved crashes occurred among 15-to 19-year-olds than would have occurred had the purchase age not been reduced to 18 years. The effect size for 18- to 19-year-olds is remarkable given the legal exceptions to the pre-1999 law and its poor enforcement. PMID- 16317198 TI - The ethics of industry experimentation using employees: the case of taste-testing pesticide-treated tobacco. AB - In the United States, companies that use their own funds to test consumer products on their employees are subject to few regulations. Using previously undisclosed tobacco industry documents, we reviewed the history of that industry's efforts to create internal guidelines on the conditions to be met before employee taste testers could evaluate cigarettes made from tobacco treated with experimental pesticides. This history highlights 2 potential ethical issues raised by unregulated industrial research: conflict of interest and lack of informed consent. To ensure compliance with accepted ethical standards, an independent federal office should be established to oversee industrial research involving humans exposed to experimental or increased quantities of ingested, inhaled, or absorbed chemical agents. PMID- 16317199 TI - Tobacco control, stigma, and public health: rethinking the relations. AB - The AIDS epidemic has borne witness to the terrible burdens imposed by stigmatization and to the way in which marginalization could subvert the goals of HIV prevention. Out of that experience, and propelled by the linkage of public health and human rights, came the commonplace assertion that stigmatization was a retrograde force.Yet, strikingly, the antitobacco movement has fostered a social transformation that involves the stigmatization of smokers. Does this transformation represent a troubling outcome of efforts to limit tobacco use and its associated morbidity and mortality; an ineffective, counterproductive, and moralizing approach that leads to a dead end; or a signal of public health achievement? If the latter is the case, are there unacknowledged costs? PMID- 16317200 TI - Junking good science: undoing Daubert v Merrill Dow through cross-examination and argument. AB - For more than 40 years, the tobacco industry prevailed in lawsuits brought by injured smokers, despite overwhelming epidemiological evidence that smoking caused lung cancer. Tobacco lawyers were able to create doubt about causation. They sought to persuade jurors that "everybody knew" smoking was harmful but "nobody knows" what causes cancer by recreating in court the scientific debate resolved by the 1964 Surgeon General's Report. The particularistic structure of jury trials combined with the law's mechanistic view of causation enables a defendant to contest virtually any claim concerning disease causation. Despite judicial efforts to eliminate "junk science" from lawsuits, a well-financed defendant may succeed in persuading jurors of the epidemiological equivalent of the proposition that the earth is flat. PMID- 16317201 TI - Key challenges to achieving health for all in an inequitable society: the case of South Africa. AB - The health inequalities in South Africa are rapidly worsening. Since 1994, the new democratic government has initiated a number of large-scale policies and programs with explicit pro-equity objectives that have improved access to health care and other social resources. However, these policies and programs have been constrained by macroeconomic policies that dictate fiscal restraint and give priority to technical rather than developmental considerations. We propose an approach to improving health for all that focuses on equity in the allocation of health resources. The implementation of pro-equity policies requires, in addition to technically efficacious interventions, both advocacy initiatives and communication with, and the involvement of, affected communities. The Cape Town Equity Gauge project is presented as one example of a response to the challenge of inequity. PMID- 16317203 TI - Tobacco industry influence on science and scientists in Germany. AB - Using tobacco industry documents, we examined how and why the tobacco industry sought to influence science and scientists in Germany as a possible factor in explaining the German opposition to stricter tobacco regulation. Smoking and health research programs were organized both separately by individual tobacco companies and jointly through their German trade organization. An extensive network of scientists and scientific institutions with tobacco industry links was developed. Science was distorted in 5 ways: suppression, dilution, distraction, concealment, and manipulation. The extent of tobacco industry influence over the scientific establishment in Germany is profound. The industry introduced serious bias that probably influenced scientific and public opinion in Germany. This influence likely undermined efforts to control tobacco use. PMID- 16317204 TI - Contextual influences on the use of health facilities for childbirth in Africa. AB - OBJECTIVES: Previous studies of maternal health-seeking behavior focused on individual- and household-level factors. We examined community-level influences on the decision to deliver a child in a health facility across 6 African countries. METHODS: Demographic and Health Survey data were linked with contextual data, and multilevel models were fitted to identify the determinants of childbirth in a health facility in the 6 countries. RESULTS: We found strong community-level influences on a woman's decision to deliver her child in a health facility. Several pathways of influence between the community and individual were identified. CONCLUSIONS: Community economic development, the climate of female autonomy, service provision, and fertility preferences all exert an influence on a woman's decision to seek care during labor, but significant community variation remains unexplained. PMID- 16317205 TI - Cost-effectiveness of free HIV voluntary counseling and testing through a community-based AIDS service organization in Northern Tanzania. AB - OBJECTIVES: We evaluated the cost-effectiveness of fee-based and free testing strategies at an HIV voluntary counseling and testing (VCT) program integrated into a community-based AIDS service organization in Moshi, Tanzania. METHODS: We waived the usual fee schedule during a 2-week free, advertised VCT campaign; analyzed the number of clients testing per day during prefree, free, and postfree testing periods; and estimated the cost-effectiveness of limited and sustained free testing strategies. RESULTS: The number of clients testing per day increased from 4.1 during the prefree testing interval to 15.0 during the free testing campaign (P<.0001) and remained significantly increased at 7.1 (P<.0001) after resumption of the standard fees. HIV seroprevalence (16.7%) and risk behaviors were unchanged over these intervals. Modeled over 1 year, the costs per infection averted with the standard fee schedule, with a 2-week free VCT campaign, and with sustained free VCT year-round were $170, $105, and $92, respectively, and the costs per disability-adjusted life year gained were $8.72, $5.40, and $4.72, respectively. CONCLUSIONS: The provision of free VCT enhances both the number of clients testing per day and its cost-effectiveness in resource-limited settings. PMID- 16317206 TI - FIDELIS--innovative approaches to increasing global case detection of tuberculosis. AB - Tuberculosis was declared a global public health emergency in 1993. In 2003, only 45% of the world's estimated new smear-positive tuberculosis cases were detected well below the 70% global case detection target set by the World Health Organization. The FIDELIS (Fund for Innovative DOTS Expansion Through Local Initiatives to Stop TB) initiative is a new global disease control initiative that has been developed to rapidly assess and implement innovative approaches to increase tuberculosis case detection. To date, 32 projects have been approved covering approximately 378 million people in 13 countries-24 (75%) of which are in the world's 6 highest-burden countries. A wide range of target populations and interventions have been incorporated into successful FIDELIS projects. The FIDELIS initiative may serve as a model to discover best practices to address other urgent global public health problems. PMID- 16317207 TI - Inequities in mental health care after health care system reform in Chile. AB - OBJECTIVES: We compared differences in mental health needs and provision of mental health services among residents of Santiago, Chile, with private and public health insurance coverage. METHODS: We conducted a cross-sectional survey of a random sample of adults. Presence of mental disorders and use of health care services were assessed via structured interviews. Individuals were classified as having public, private, or no health insurance coverage. RESULTS: Among individuals with mental disorders, only 20% (95% confidence interval [CI]=16%, 24%) had consulted a professional about these problems. A clear mismatch was found between need and provision of services. Participants with public insurance coverage exhibited the highest prevalence of mental disorders but the lowest rates of consultation; participants with private coverage exhibited exactly the opposite pattern. After adjustment for age, income, and severity of symptoms, private insurance coverage (odds ratio [OR]=2.72; 95% CI=1.6, 4.6) and higher disability level (OR=1.27, 95% CI=1.1, 1.5) were the only factors associated with increased frequency of mental health consultation. CONCLUSIONS: The health reforms that have encouraged the growth of the private health sector in Chile also have increased risk segmentation within the health system, accentuating inequalities in health care provision. PMID- 16317208 TI - Decreasing socioeconomic inequalities and increasing health inequalities in Spain: a case study. AB - OBJECTIVES: We examined the evolution of income inequalities and health inequalities in Spain from the time of the country's entry into the European Union. METHODS: We estimated distributions of provincial income and household income, relations of provincial income with mortality and disability, and relations of household income with disability in 1984-1986 and 1999-2001. RESULTS: Inequalities in average provincial income and household income were lower in 2000 than in 1985. Differences in mortality and disability according to income were greater in 2000 than in 1985, in both absolute and relative terms, except for differences in mortality among individuals aged 25 to 44 years. In most cases, differences in mortality from leading causes of death and differences in major types of disabilities were also greater in 2000. CONCLUSIONS: Our results show that redistribution of income might achieve greater social justice but probably does not lead to reduced health inequalities, despite observed improvements in material circumstances as well as in most health indicators among disadvantaged population groups. PMID- 16317209 TI - The roles of teaching hospitals, insurance status, and race/ethnicity in receipt of adjuvant therapy for regional-stage breast cancer in Florida. AB - OBJECTIVES: We examined the roles of teaching hospitals, insurance status, and race/ ethnicity in women's receipt of adjuvant therapy for regional-stage breast cancer. METHODS: Data were taken from the Florida Cancer Data System for cases diagnosed from July 1997 to December 2000. We evaluated the impact of health insurance status and hospital type on use of adjuvant therapy (after adjustment for age, race/ethnicity, and marital status). Interaction terms for hospital type, insurance status, and race/ethnicity were entered in each model. RESULTS: Teaching facilities diagnosed 12.5% of the cases; however, they cared for a disproportionate percentage (21.3%) of uninsured and Medicaid-insured women. Among women who received adjuvant chemotherapy only, those diagnosed in teaching hospitals were more likely than those diagnosed in nonteaching hospitals to receive therapy regardless of insurance status or race/ethnicity. Among women who received chemotherapy with or without hormonal therapy, Hispanics were more likely than White non-Hispanic women to receive therapy, whereas women with private insurance or Medicare were less likely than uninsured and Medicaid insured women to receive this type of therapy. CONCLUSIONS: Teaching facilities play an important role in the diagnosis and treatment of regional-stage breast cancer among Hispanics, uninsured women, and women insured by Medicaid. PMID- 16317210 TI - Structure and functions of state public health agencies. AB - OBJECTIVES: We assessed the structure and functions of state health departments throughout the United States and compared our findings with those from a previous national assessment conducted in 1990. METHODS: In 2001, we sent a survey to the state health officers of all 50 states. The survey asked about the structure and functions of the state health agency. RESULTS: The survey was completed by state health officers from 47 states (a 94% response rate). More than half of the states responding had a freestanding state public health agency and a state board or council of health. Forty-four percent had a regional or district structure. Although some traditional public health functions have been curtailed, important new public health functions have emerged since 1990. CONCLUSIONS: Our current findings confirm core changes in the structure and functions of state public health systems over the past decade and emphasize the need for more research into these systems to maximize their organizational performance. PMID- 16317211 TI - Cardiovascular disease and associated risk factors in Cuba: prospects for prevention and control. AB - OBJECTIVES: An adequate description of the trends in cardiovascular disease (CVD) is not available for most of the developing world. Cuba provides an important exception, and we sought to use available data to offer insights into the changing patterns of CVD there. METHODS: We reviewed Cuban public health statistics, surveys, and reports of health services. RESULTS: CVD has been the leading cause of death since 1970. A 45% reduction in heart disease deaths was observed from 1970 to 2002; the decline in stroke was more limited. There are moderate prevalences of all major risk factors. CONCLUSIONS: The Cuban medical care system has responded vigorously to the challenge of CVD; levels of control of hypertension are the highest in the world. Nonindustrialized countries can decisively control CVD. PMID- 16317212 TI - Urban-rural inequalities in ischemic heart disease in Scotland, 1981-1999. AB - OBJECTIVES: We sought to describe the pattern and magnitude of urban-rural variation in ischemic heart disease (IHD) in Scotland and to examine the associations among IHD health indicators, level of rurality, and degree of socioeconomic deprivation. METHODS: We used routine population and health data on the population aged 40-74 years between 1981 and 1999 and living in 826 small areas (average population=5600) in Scotland. Three IHD health indicators mortality rates (deaths per 100,000 population), rates of continuous hospital stays (discharges per 100,000 population), and rates of mortality in the hospital or within 28 days of discharge (MH+) were analyzed with multilevel Poisson models. A 4-level rurality classification was used: urban areas, remote small towns, accessible rural areas, and remote rural areas. RESULTS: Rates of mortality, continuous hospital stays, and MH+ increased with area socioeconomic deprivation. After adjustment for population age, gender, and deprivation, the relative risk of IHD mortality in remote rural areas was similar to that of urban areas in 1981; the relative risk of a continuous hospital stay was significantly lower (relative risk [RR] = 0.70; 95% confidence interval [CI] = 0.64, 0.76) and the relative risk of MH+ was higher (RR=1.18; 95% CI=1.04, 1.35) in remote rural areas. Mortality and MH+ declined for all ruralities over time. However, MH+ remains highest in remote rural areas and remote towns. CONCLUSIONS: Low standardized ratios of IHD continuous hospital stays and mortality in remote rural areas mask health problems among rural populations. Although absolute and relative differences between urban and rural rates of MH+ have diminished, the relative risk of MH+ remains high in remote rural areas. PMID- 16317213 TI - Individual and contextual determinants of domestic violence in North India. AB - OBJECTIVES: We examined individual- and community-level influences on domestic violence in Uttar Pradesh, North India. METHODS: Multilevel modeling was used to explore domestic violence outcomes among a sample of 4520 married men. RESULTS: Recent physical and sexual domestic violence was associated with the individual level variables of childlessness, economic pressure, and intergenerational transmission of violence. A community environment of violent crime was associated with elevated risks of both physical and sexual violence. Community-level norms concerning wife beating were significantly related only to physical violence. CONCLUSIONS: Important similarities as well as differences were evident in risk factors for physical and sexual domestic violence. Higher socioeconomic status was found to be protective against physical but not sexual violence. Our results provide additional support for the importance of contextual factors in shaping women's risks of physical and sexual violence. PMID- 16317214 TI - Integrating behavioral and social science research into microbicide clinical trials: challenges and opportunities. AB - It has been argued that rigid thinking about the types and progression of research needed to evaluate health promotion interventions has stymied the process by which research is translated to action. This argument is particularly salient in the field of HIV/AIDS prevention. We examined microbicide research and identified challenges that obstruct the integration of clinical trial and behavioral and social science research, thereby reinforcing linear programs of research. We found that behavioral and social science research can both support microbicide clinical trial performance and anticipate the information most needed for a rapid and successful introduction of future microbicide products. PMID- 16317215 TI - Environmental-structural interventions to reduce HIV/STI risk among female sex workers in the Dominican Republic. AB - OBJECTIVES: We assessed the effectiveness of 2 environmental-structural interventions in reducing risks of HIV and sexually transmitted infections (STIs) among female sex workers in the Dominican Republic. METHODS: Two intervention models were implemented over a 1-year period: community solidarity in Santo Domingo and solidarity combined with government policy in Puerto Plata. Both were evaluated via preintervention-postintervention cross-sectional behavioral surveys, STI testing and participant observations, and serial cross-sectional STI screenings. RESULTS: Significant increases in condom use with new clients (75.3% 93.8%; odds ratio [OR]=4.21; 95% confidence interval [CI]=1.55, 11.43) were documented in Santo Domingo. In Puerto Plata, significant increases in condom use with regular partners (13.0%-28.8%; OR=2.97; 95% CI=1.33, 6.66) and reductions in STI prevalence (28.8%-16.3%; OR = 0.50; 95% CI = 0.32, 0.78) were documented, as were significant increases in sex workers' verbal rejections of unsafe sex (50.0% 79.4%; OR=3.86; 95% CI=1.96, 7.58) and participating sex establishments' ability to achieve the goal of no STIs in routine monthly screenings of sex workers (OR=1.17; 95% CI=1.12, 1.22). CONCLUSIONS: Interventions that combine community solidarity and government policy show positive initial effects on HIV and STI risk reduction among female sex workers. PMID- 16317216 TI - Pattern of diabetic foot at Koja Regional General Hospital, Jakarta, from 1999 to 2004. AB - AIM: To determine the prevalence of diabetic foot according to severity of disease, causing microorganism, resistance pattern of microorganism, and blood glucose level of diabetes mellitus (DM) patients at Koja Regional General Hospital, to provide additional information to improve patient follow-up. METHODS: This study is designed as a descriptive cross-sectional study. The population in this study consists of patients with diabetes mellitus patients admitted and treated at the internal medicine outpatient clinic of Koja Regional General Hospital from January 1999 to December 2004. RESULTS: We obtained a prevalence of diabetic foot according to the causal microorganisms as follows: Pseudomonas (40.9%), followed by Staphylococcus (29.5%). The prevalence of diabetic foot according to the blood glucose groups were: 9.5% having random blood glucose concentration of 150-200 mg/dl, 9.3% 201-250 mg/dl, 30% 251-300 mg/dl, and 30% >300 mg/dl. According to Wagner classification, most cases were classified as 0 degree, highly effective antibiotics treatment followed by fifth degree at 13.5%. In addition, the Pseudomonas were Cefotaxim 53.8% and Ceftriaxon 41.5%. CONCLUSION: According to the results of this study, the most common cause of diabetic foot is Pseudomonas, and the effective antibiotics treatment is Ceftriaxone and Cefotaxim. The prevalence of diabetic foot has increased along with increased blood glucose level, and according to Wagner classification, 0 degree was the most common, but fifth degree was also higher than 2nd to 4th degree. PMID- 16317217 TI - Comparison of tumor density by CT scan based on histologic type in lung cancer patients. AB - AIM: To investigate the differences of tumor density of lung cancer by CT scan based on histopathologic type of cancer cell. METHODS: Subjects were lung cancer patients at Dr. Kariadi hospital Semarang between January 2002 and December 2003. Diagnosis was made based on clinical symptoms, histologic examination and lateral chest radiograph, and thorax CT scan. Density of tumor was measured within area which had most enhancements before and after contrast administration. Then, it was followed by transthoraxic fine needle aspiration (TNFNAB) and histopathologic examination. Study design was a cross sectional study. Inclusion criteria were primary lung cancer and exclusion criteria were non lung cancer, metastatic cancer, histopathologic non interpreted. Estimated number of samples was 41 patients. Density value before and after contrast were analyzed using Wilcoxon signed rank test. Difference of tumor density for each type of cancer cell was analyzed using chi-square test with significant level at p < 0.05 and confidence interval of 95%. RESULTS: Forty-one patients with lung cancer were enrolled in this study. There were 30 males (73.2%) and 11 females (26.8%). Histopathologic types of cell cancer were epidermoid in 17 patients (14.5%), adenocarcinoma 8 patients (19.5%), large cell 7 patients ( 17.1%), small cell 7 patients ( 17.1%) and undifferentiated cell 2 patients ( 4.5%). Mean of increasing HU before and after contrast in epidermoid carcinoma was 26.8 (10.4) HU, adenocarcinoma 17.2 (7.1) HU, large cell 8.7 (4.5) HU, small cell 9.9(4.1) HU and undifferentiated cell 23.5 (1.5) HU with overall increase of all cancer cell types was 17.3(9.2) HU, p < 0.001. Histopathologic type of non small cell lung carcinoma (NSCLC) with density < 60 HU was found in 30 patients (73.2%) and small cell lung carcinoma with density > 60 HU was found in 5 patient (12.2%), p=0.01. CONCLUSION: There was evident increase of density in all histopathologic type of cell cancer, p < 0.0001, with mean of density after contrast administration was 17.2 (7.1). There was significant difference between NSCLC and SCLC group and tumor density > 60 HU and < 60 HU, p=0.01. PMID- 16317218 TI - The efficacy of granisetron for cancer patients undergoing platinum-based chemotherapy: comparison of 1 miligram versus 3 miligram doses in preventing nausea and vomiting. AB - AIM: To compare the efficacy of anti-emetic and prophylactic effects of 1 milligram (mg) versus 3 mg granisetron in cancer patients. METHODS: In this double blind, randomized, parallel study, 2-dose regimens of intra venous (IV) granisetron were evaluated in 39 cancer patients who were treated with platinum based chemotherapy. Patients who met the inclusion criteria were randomized to receive granisetron 1 mg IV plus dexamethasone 20 mg (group A) or granisetron 3 mg iv plus dexamethasone 20 mg (group B). A questionnaire was used to evaluate the anti-emetic effects of granisetron. RESULTS: Subjects consisted of 31 men and 8 women. In group A (19 patients) 57.9% showed complete response from vomiting, 10.5% of major response, and 31.6% of failure to anti-emetic therapy. There were 47.4% of patients free from nausea and 52.6% complained of nausea (mild, moderate, and severe nausea). Among group B (20 patients), 90% showed complete response from vomiting, 5% of major response, and 5% of failure to anti-emetic therapy. Eighty percent of patients were free from nausea, while 15% complained of mild nausea and 5% of moderate nausea. The differences were statistically significant for vomiting (p = 0.02) as well as for nausea (p = 0.03). CONCLUSION: Intravenous granisetron 3 mg has better efficacy than granisetron 1 mg in preventing cisplatin- induced acute emesis. PMID- 16317219 TI - Troponin in heart disorder and chronic renal failure. PMID- 16317220 TI - Insomnia in geriatrics. PMID- 16317221 TI - Osteoarticular tuberculosis. PMID- 16317222 TI - Epidemiology of hepatitis B. PMID- 16317223 TI - Stratification of risk factors in venous thromboembolism: use of the D-dimer test. PMID- 16317224 TI - Characterization of four-class motor imagery EEG data for the BCI-competition 2005. AB - To determine and compare the performance of different classifiers applied to four class EEG data is the goal of this communication. The EEG data were recorded with 60 electrodes from five subjects performing four different motor-imagery tasks. The EEG signal was modeled by an adaptive autoregressive (AAR) process whose parameters were extracted by Kalman filtering. By these AAR parameters four classifiers were obtained, namely minimum distance analysis (MDA)--for single channel analysis, and linear discriminant analysis (LDA), k-nearest-neighbor (kNN) classifiers as well as support vector machine (SVM) classifiers for multi channel analysis. The performance of all four classifiers was quantified and evaluated by Cohen's kappa coefficient, an advantageous measure we introduced here to BCI research for the first time. The single-channel results gave rise to topographic maps that revealed the channels with the highest level of separability between classes for each subject. Our results of the multi-channel analysis indicate SVM as the most successful classifier, whereas kNN performed worst. PMID- 16317225 TI - Multi-site incorporation of bioactive matrices into MEMS-based neural probes. AB - Methods are presented to incorporate polymer-based bioactive matrices into micro fabricated implantable microelectrode arrays. Using simple techniques, hydrogels infused with bioactive molecules are deposited within wells in the substrate of the device. This method allows local drug delivery without increasing the footprint of the device. In addition, each well can be loaded individually, allowing spatial and temporal control over diffusion gradients in the microenvironment of the implanted neural interface probe. In vivo testing verified the following: diffusion of the bioactive molecules, integration of the bioactive molecules with the intended neural target and concurrent extracellular recording using nearby electrodes. These results support the feasibility of using polymer gels to deliver bioactive molecules to the region close to microelectrode shanks. This technique for microdrug delivery may serve as a means to intervene with the initial phases of the neuroinflammatory tissue response to permanently implanted microelectrode arrays. PMID- 16317226 TI - Improving correct switching rates in a 'hands-free' environmental control system. AB - One potential negative impact on the quality of life of a spinal cord injured person is the loss of the ability to control devices in their immediate environment. Consequently, research and development has been conducted on technology designed to restore some measure of independence by providing means of control over these devices. A previous assistive device using changes in brain signals from eye closure as its switching system was created. Brain signals were processed using spectral analysis and although this was a successful technique, there were limitations that resulted in higher than desired switching errors. This paper presents results of an alternative method for processing brain signals as the basis for switching, called fractal dimension. In comparison to the spectral technique, the fractal dimension technique was successful in reducing the number of false positive and false negative errors. Additionally, it eliminated the need for a baseline setup for this system. This suggests that fractal dimension is a potentially viable method for analysing brain signals for use in assistive control systems. PMID- 16317227 TI - Development of a cortical visual neuroprosthesis for the blind: the relevance of neuroplasticity. AB - Clinical applications such as artificial vision require extraordinary, diverse, lengthy and intimate collaborations among basic scientists, engineers and clinicians. In this review, we present the state of research on a visual neuroprosthesis designed to interface with the occipital visual cortex as a means through which a limited, but useful, visual sense could be restored in profoundly blind individuals. We review the most important physiological principles regarding this neuroprosthetic approach and emphasize the role of neural plasticity in order to achieve desired behavioral outcomes. While full restoration of fine detailed vision with current technology is unlikely in the immediate near future, the discrimination of shapes and the localization of objects should be possible allowing blind subjects to navigate in a unfamiliar environment and perhaps even to read enlarged text. Continued research and development in neuroprosthesis technology will likely result in a substantial improvement in the quality of life of blind and visually impaired individuals. PMID- 16317228 TI - Beyond sensory substitution--learning the sixth sense. AB - Rapid advances in neuroscience have sparked numerous efforts to study the neural correlate of consciousness. Prominent subjects include higher sensory area, distributed assemblies bound by synchronization of neuronal activity and neurons in specific cortical laminae. In contrast, it has been suggested that the quality of sensory awareness is determined by systematic change of afferent signals resulting from behaviour and knowledge thereof. Support for such skill-based theories of perception is provided by experiments on sensory substitution. Here, we pursue this line of thought and create new sensorimotor contingencies and, hence, a new quality of perception. Adult subjects received orientation information, obtained by a magnetic compass, via vibrotactile stimulation around the waist. After six weeks of training we evaluated integration of the new input by a battery of tests. The results indicate that the sensory information provided by the belt (1) is processed and boosts performance, (2) if inconsistent with other sensory signals leads to variable performance, (3) does interact with the vestibular nystagmus and (4) in half of the experimental subjects leads to qualitative changes of sensory experience. These data support the hypothesis that new sensorimotor contingencies can be learned and integrated into behaviour and affect perceptual experience. PMID- 16317229 TI - A wavelet-based time-frequency analysis approach for classification of motor imagery for brain-computer interface applications. AB - Electroencephalogram (EEG) recordings during motor imagery tasks are often used as input signals for brain-computer interfaces (BCIs). The translation of these EEG signals to control signals of a device is based on a good classification of various kinds of imagination. We have developed a wavelet-based time-frequency analysis approach for classifying motor imagery tasks. Time-frequency distributions (TFDs) were constructed based on wavelet decomposition and event related (de)synchronization patterns were extracted from symmetric electrode pairs. The weighted energy difference of the electrode pairs was then compared to classify the imaginary movement. The present method has been tested in nine human subjects and reached an averaged classification rate of 78%. The simplicity of the present technique suggests that it may provide an alternative method for EEG based BCI applications. PMID- 16317230 TI - Dilation of the oropharynx via selective stimulation of the hypoglossal nerve. AB - The functional effects of selective hypoglossal nerve (HG) stimulation with a multi-contact peripheral nerve electrode were assessed using images of the upper airways and the tongue in anesthetized beagles. A biphasic pulse train of 50 Hz frequency and 2 s duration was applied through each one of the tripolar contact sets of the nerve electrode while the pharyngeal images were acquired into a computer. The stimulation current was limited to 20% above the activation threshold for maximum selectivity. The images showed that various contact sets could generate several different activation patterns of the tongue muscles resulting in medial and/or lateral dilation and closing of the airways at the tongue root. Some of these patterns translated into an increase in the oropharyngeal size while others did not have any effect. The pharyngeal sizes were not statistically different during stimulation either between the two different positions of the head (30 degrees and 60 degrees), or when the lateral contacts were compared with the medial ones. The contacts that had the least effect generated an average of 53 +/- 15% pharyngeal dilation relative to the best contacts, indicating that the results are marginally sensitive to the contact position around the HG nerve trunk. These results suggest that selective HG nerve stimulation can be a useful technique to produce multiple tongue activation patterns that can dilate the pharynx. This may in turn increase the size of the patient population who can benefit from HG nerve stimulation as a treatment method for obstructive sleep apnea. PMID- 16317231 TI - Biomechanical analysis of silicon microelectrode-induced strain in the brain. AB - The ability to successfully interface the brain to external electrical systems is important both for fundamental understanding of our nervous system and for the development of neuroprosthetics. Silicon microelectrode arrays offer great promise in realizing this potential. However, when they are implanted into the brain, recording sensitivity is lost due to inflammation and astroglial scarring around the electrode. The inflammation and astroglial scar are thought to result from acute injury during electrode insertion as well as chronic injury caused by micromotion around the implanted electrode. To evaluate the validity of this assumption, the finite element method (FEM) was employed to analyze the strain fields around a single Michigan Si microelectrode due to simulated micromotion. Micromotion was mimicked by applying a force to the electrode, fixing the boundaries of the brain region and applying appropriate symmetry conditions to nodes lying on symmetry planes. Characteristics of the deformation fields around the electrode including maximum electrode displacement, strain fields and relative displacement between the electrode and the adjacent tissue were examined for varying degrees of physical coupling between the brain and the electrode. Our analysis demonstrates that when physical coupling between the electrode and the brain increases, the micromotion-induced strain of tissue around the electrode decreases as does the relative slip between the electrode and the brain. These results support the use of neuro-integrative coatings on electrode arrays as a means to reduce the micromotion-induced injury response. PMID- 16317232 TI - Separation of spinal cord motor signals using the FastICA method. AB - Evoked motor signals descending down the corticospinal tract can be recorded selectively with multi-contact electrodes from the spinal cord surface. This method of extracting motor signals from the spinal cord may provide a means of communication for people with spinal cord injury. The information rate obtained with such an interface will improve if the separation of neural channels can be increased. In this study, the feasibility of increasing the channel separation was investigated using the blind source separation (BSS) technique. Neural signals recorded with multi-contact surface electrodes were treated as a linear mixture of independent neural sources located inside the spinal cord. Principal component analysis (PCA) was applied to estimate the dimensionality of the raw signals, and then the fixed-point FastICA algorithm was used to separate the primary neural sources from the secondary (smaller) ones. In all trials but one, the separation between the neural channels has increased by eliminating the secondary sources. These results suggest that the information rate of a spinal cord interface can be improved by separating the neural recordings into their independent components and selecting the ones with the largest distance between them. Comparison of independent component analysis (ICA) and PCA reveals that ICA performs better in this application. PMID- 16317233 TI - Visualization of the intact interface between neural tissue and implanted microelectrode arrays. AB - This research presents immunohistochemical strategies for assessing the interactions at the immediate interface between micro-scale implanted devices and the surrounding brain tissue during inflammatory astrogliotic reactions. This includes preparation, microscopy and analysis techniques for obtaining images of the intimate contact between neural cells and the surface of implantable micro electromechanical systems (MEMS) devices. The ability to visualize the intact interface between an implant and the surrounding tissue allows researchers to examine tissue that is unchanged from its native implanted state. Conversely, current popular techniques involve removing the implant. This tends to cause damage to the tissue immediately surrounding the implant and can hinder one's ability to differentiate inflammatory responses to the implant versus physical damage occurring from removal of the implant from the tissue. Due to advances in microscopy and staining techniques, it is now possible to visualize the intact tissue-implant interface. This paper presents the development of imaging techniques for visualizing the intact interface between neural tissue and implanted devices. This is particularly important for understanding both the acute and chronic neuroinflammatory responses to devices intended for long-term use in a prosthetic system. Non-functional, unbonded devices were imaged in vitro and in vivo at different times post-implantation via a range of techniques. Using these techniques, detailed interactions could be seen between delicate cellular processes and the electrode surface, which would have been destroyed using conventional histology processes. PMID- 16317234 TI - A finite-element model of the mechanical effects of implantable microelectrodes in the cerebral cortex. AB - The viability of chronic neural microelectrodes for electrophysiological recording and stimulation depends on several factors, including the encapsulation of the implant by a reactive tissue response. We postulate that mechanical strains induced around the implant site may be one of the leading factors responsible for the sustained tissue response in chronic implants. The objectives of this study were to develop a finite-element model of the probe-brain tissue interface and analyze the effects of tethering forces, probe-tissue adhesion and stiffness of the probe substrate on the interfacial strains induced around the implant site. A 3D finite-element model of the probe-brain tissue microenvironment was developed and used to simulate interfacial strains created by 'micromotion' of chronically implanted microelectrodes. Three candidate substrates were considered: (a) silicon, (b) polyimide and (c) a hypothetical 'soft' material. Simulated tethering forces resulted in elevated strains both at the tip and at the sharp edges of the probe track in the tissue. The strain fields induced by a simulated silicon probe were similar to those induced by a simulated polyimide probe, albeit at higher absolute values for radial tethering forces. Simulations of poor probe-tissue adhesion resulted in elevated strains at the tip and delamination of the tissue from the probe. A tangential tethering force results in 94% reduction in the strain value at the tip of the polyimide probe track in the tissue, whereas the simulated 'soft' probe induced two orders of magnitude smaller values of strain compared to a simulated silicon probe. The model results indicate that softer substrates reduce the strain at the probe tissue interface and thus may also reduce tissue response in chronic implants. PMID- 16317235 TI - Poly-HEMA as a drug delivery device for in vitro neural networks on micro electrode arrays. AB - Delivery of pharmacological agents in vitro can often be a difficult, time consuming and costly process. In this paper, we describe an economical method for in vitro delivery using a hydrogel of poly hydroxyethyl methacrylate (PHEMA) that can absorb up to 50% of its weight of any water-solubilized pharmacological agent. This agent will then passively diffuse into surrounding media upon application in vitro. An in vitro test of PHEMA as a drug delivery device was conducted using dissociated rat-cortical neurons cultured on micro-electrode arrays. These micro-electrode arrays permit the real-time measurement of neural activity at 60 different sites across a network of neurons. Neural activity was compared during the application of PHEMA saturated with cell culture media and PHEMA saturated with bicuculline, a widely used pharmacological agent with stereotypical effects on neural activity patterns. Application of PHEMA saturated with bicuculline produced a gradual increase in concentration in vitro. When the minimum effective concentration of bicuculline was reached, which was found to be 0.59 microM using the diffusion properties of PHEMA, it produced the rapid almost periodic synchronized bursting characteristically associated with this agent. In contrast, the application of PHEMA saturated in culture media alone had no effect on neural activity reinforcing its inherent inert properties. Since PHEMA is nontoxic, can be molded into a variety of shapes, quickly manufactured in any laboratory and is inexpensive to produce, the material represents a promising alternative to drug delivery systems on the market today. PMID- 16317236 TI - Steady-state visual evoked potential (SSVEP)-based communication: impact of harmonic frequency components. AB - Brain-computer interfaces (BCIs) can be realized on the basis of steady-state evoked potentials (SSEPs). These types of brain signals resulting from repetitive stimulation have the same fundamental frequency as the stimulation but also include higher harmonics. This study investigated how the classification accuracy of a 4-class BCI system can be improved by incorporating visually evoked harmonic oscillations. The current study revealed that the use of three SSVEP harmonics yielded a significantly higher classification accuracy than was the case for one or two harmonics. During feedback experiments, the five subjects investigated reached a classification accuracy between 42.5% and 94.4%. PMID- 16317237 TI - Classification of the intention to generate a shoulder versus elbow torque by means of a time-frequency synthesized spatial patterns BCI algorithm. AB - In this paper, we attempt to determine a subject's intention of generating torque at the shoulder or elbow, two neighboring joints, using scalp electroencephalogram signals from 163 electrodes for a brain-computer interface (BCI) application. To achieve this goal, we have applied a time-frequency synthesized spatial patterns (TFSP) BCI algorithm with a presorting procedure. Using this method, we were able to achieve an average recognition rate of 89% in four healthy subjects, which is comparable to the highest rates reported in the literature but now for tasks with much closer spatial representations on the motor cortex. This result demonstrates, for the first time, that the TFSP BCI method can be applied to separate intentions between generating static shoulder versus elbow torque. Furthermore, in this study, the potential application of this BCI algorithm for brain-injured patients was tested in one chronic hemiparetic stroke subject. A recognition rate of 76% was obtained, suggesting that this BCI method can provide a potential control signal for neural prostheses or other movement coordination improving devices for patients following brain injury. PMID- 16317238 TI - Current density distributions, field distributions and impedance analysis of segmented deep brain stimulation electrodes. AB - Deep brain stimulation (DBS) electrodes are designed to stimulate specific areas of the brain. The most widely used DBS electrode has a linear array of 4 cylindrical contacts that can be selectively turned on depending on the placement of the electrode and the specific area of the brain to be stimulated. The efficacy of DBS therapy can be improved by localizing the current delivery into specific populations of neurons and by increasing the power efficiency through a suitable choice of electrode geometrical characteristics. We investigated segmented electrode designs created by sectioning each cylindrical contact into multiple rings. Prototypes of these designs, made with different materials and larger dimensions than those of clinical DBS electrodes, were evaluated in vitro and in simulation. A finite element model was developed to study the effects of varying the electrode characteristics on the current density and field distributions in an idealized electrolytic medium and in vitro experiments were conducted to measure the electrode impedance. The current density over the electrode surface increased towards the edges of the electrode, and multiple edges increased the non-uniformity of the current density profile. The edge effects were more pronounced over the end segments than over the central segments. Segmented electrodes generated larger magnitudes of the second spatial difference of the extracellular potentials, and thus required lower stimulation intensities to achieve the same level of neuronal activation as solid electrodes. For a fixed electrode conductive area, increasing the number of segments (edges) decreased the impedance compared to a single solid electrode, because the average current density over the segments increased. Edge effects played a critical role in determining the current density distributions, neuronal excitation patterns, and impedance of cylindrical electrodes, and segmented electrodes provide a means to increase the efficiency of DBS. PMID- 16317239 TI - Mechanical trauma induces immediate changes in neuronal network activity. AB - During a traumatic insult to the brain, tissue is subjected to large stresses at high rates which often surpass cellular thresholds leading to cell dysfunction or death. The acute response of neurons to a mechanical trauma, however, is poorly understood. Plasma membrane disruption may be the earliest cellular outcome from a mechanical trauma. The increase in membrane permeability due to such disruptions may therefore play an important role in the initiation of deleterious cascades following brain injury. The immediate consequences of an increase in plasma membrane permeability on the electrophysiological behavior of a neuronal network exposed to the trauma have not been elucidated. We have developed an in vitro model of traumatic brain injury (TBI) that utilizes a novel device capable of applying stress at high rates to neuronal cells cultured on a microelectrode array. The mechanical insult produced by the device caused a transient increase in neuronal plasma membrane permeability, which subsided after 10 min. We were able to monitor acute spontaneous electrophysiological activity of injured cultures for at least 10 min following the insult. Firing frequency, average burst interval and spikes within burst were assessed before and after injury. The electrophysiological responses to the insult were heterogeneous, although an increase in burst intervals and in the variability of the assessed parameters were common. This study provides a multi-faceted approach to elucidate the role of neuronal plasma membrane disruptions in TBI and its functional consequences. PMID- 16317240 TI - Veno-active drugs in the management of chronic venous disease. An international consensus statement: current medical position, prospective views and final resolution. AB - BACKGROUND: Veno-active drugs (VAD) have effects on edema and symptoms related to chronic venous disease (CVD), especially so-called venous pain. VAD's effectiveness, although well established, is regularly debated. OBJECTIVE: Our purpose was to select all randomized controlled trials (RCTs) and meta-analyses devoted to VAD and symptoms in CVD, to submit them to a group of international experts in CVD and to vote with secrete ballot to determine the level of efficacy of each drug, according to EBM (Evidence-Based Medicine) rules and critical analysis. METHODS: Publications in any language devoted to VAD and venous symptoms were searched for in different databanks and submitted to the experts prior to the meeting. RESULTS: 83 papers were analyzed, including 72 RCTs or meta analyses. Experts determined the level of EBM of each drug, according to the literature and personal experience, using 3 levels of recommendation, A, B and C (from large RCTs to non-randomized trials). CONCLUSIONS: VAD are effective and may be applied in CVD when symptomatic, from C0s to C6s. However, etiological treatment of venous reflux and venous hypertension has always priority. In some cases VAD may replace compression and/or complement its effects. If respecting these prerequisites, VAD are safe and effective. PMID- 16317241 TI - The fluidity of blood in African elephants (Loxodonta africana). AB - The large cellular volume of erythrocytes and the increased plasma concentration of proteins in elephants are factors which potentially affect blood rheology adversely. To verify blood rheology, routine hemorheologic variables were analyzed in four African elephants (Loxodonta africana), housed in the zoo of Vienna. Whole blood viscosity at three different shear rates (WBV at low shear rate: WBV 0.7 s(-1) and WBV 2.4 s(-1); WBV at high shear rate: WBV 94 s(-1) done by LS30, Contraves) and erythrocyte aggregation (aggregation indices AI by LS30; aggregation indices M0, M1 by Myrenne aggregometer) were high (WBV 94 s(-1): 5.368 (5.246/5.648); WBV 2.4 s(-1): 16.291 (15.605/17.629); WBV 0.7 s(-1): 28.28 (25.537/32.173) mPa s; AI 2.4 s(-1): 0.25 (0.23/0.30); AI 0.7 s(-1): 0.24 (0.23/0.28); M0: 7.8 (6.4/8.4); M1: 30.2 (25/31)). Plasma viscosity (PV) was increased as well (1.865 (1.857/1.912) mPa s) compared to other mammalian species. These parameters would indicate a decrease in blood fluidity in elephants. However, erythrocyte rigidity (LORCA, Mechatronics) was decreased, which in contrast, has a promotive effect on peripheral perfusion. Blood rheology of the elephants was determined by a high whole blood and plasma viscosity as the result of pronounced erythrocyte aggregation and high plasma protein concentration. Thus, in the terminal vessels the resistance to flow will be increased. The large erythrocytes, which might impede blood flow further due to geometrical reasons, however, had a pronounced flexibility. We conclude that the effect of the increased inner resistance to peripheral blood flow was counteracted by the decreased rigidity of the erythrocytes to enable an adequate blood flow in African elephants. PMID- 16317242 TI - Analysis on the erythrocyte shape changes using wavelet transforms. AB - The mechanics of red blood cell shape changes under normal and deformed conditions are analyzed using wavelet based approach. Images of intact and deformed human red blood cells obtained from normal adults are subject to morphological image processing and the corresponding shape descriptions at two different levels of approximations using different wavelet functions are analyzed. The results demonstrate that using wavelets it is possible to classify normal and deformed red blood cell shapes. Uniform and consistent results are obtained for cells with similar shapes, for all chosen wavelet functions. The variation indices are significant (p < 0.005) for all the chosen wavelet functions at both the approximation levels. Further it seems that this approach could be useful for identifying closely identical cell shapes. As cell shape deformations are significant in describing the flow behavior in micro or macro vessels the study seems to be clinically relevant. The methodologies, algorithms and observations based on wavelet based analysis are discussed in detail. PMID- 16317243 TI - Glycaemic control influences peripheral blood flow and haemorheological variables in patients with diabetes mellitus. AB - Since persistent uncontrolled hyperglycaemia predisposes to vascular complications in diabetics, this study aimed at assessing the relationship of glycaemic control to plasma fibrinogen concentration, relative plasma viscosity and ankle arterial blood flow in diabetic patients with (N = 28) and without neuropathy (N = 34) compared with non-diabetic controls (N = 21). Glycaemic control was determined by total glycated haemoglobin (GHb) levels. Patients were placed into three categories of glycaemic control, namely good (GHb 4 -< 8%), fair (GHb 8-12%) and poor (GHb > 12%).Compared with non-diabetics, blood flow was significantly higher (p < 0.05) in patients with good but not poor glycaemic control. Fibrinogen was significantly higher in patients with fair and poor glycaemic control than in non-diabetic subjects (p < 0.05). In non-neuropathic patients, viscosity was higher (p < 0.05) in those with fair control and significantly different (p < 0.05) between those with fair and poor control. The results suggest that the initial vasodilatation in the periphery is attenuated by poor glycaemic control, contributing to the decrease in ankle arterial blood flow as a consequence of the simultaneous increase in plasma fibrinogen and viscosity. These adverse changes may contribute to the development of the diabetic foot. PMID- 16317244 TI - Alterations in hemorheological properties of erythrocytes in nude mice with erythroleukemia and the treatment effects of etoposide. AB - The purpose of this study was to examine the changes of hemorheological properties of erythrocytes in the nude mice with erythroleukemia and the treatment effects of etoposide (VP16). Thirty mice were randomly divided into three groups: the control group (C group), injected with 1 ml saline solution, the MEL group (M group) injected with 1 ml MEL (murine erythroleukemia cell line) and the MEL + VP16 group (V group) injected with 1 ml MEL and from the 8th day after injection, 20 microl VP16 (1 microg/microl) was injected intraperitoneally every five days. One week after MEL injection, erythroblastic cells increased in the bone marrow and proerythroblasts were found in the peripheral blood, suggesting that erythroleukemia was induced. Abnormalities were also found in spleens and livers later. At around twenty days after injection, the mice in M group died and about four weeks after injection, the mice in V group also died. Compared with C group, the hemorheological indexes [the deformation index DI, orientation index (DI(or)), and the small deformation index (DI(d))], electrophoretic mobility, membrane fluidity as well as osmotic fragility of red blood cells (RBC) in M and V groups changed significantly. But after VP16 administration, the changes of above parameters in V group were less significant than those of M group. The results above suggested that intraperitoneal injection of MEL cells could cause erythroleukemia in nude mice, VP16 could alleviate the erythroleukemia symptom and improve the hemorheological properties, and could prolong V group nude mice survival. PMID- 16317245 TI - Modulation of density-fractionated RBC deformability by nitric oxide. AB - The role of nitric oxide (NO) in maintaining normal mechanical behavior of red blood cell (RBC) has been previously demonstrated. The effects of NO donor and NOS inhibitor on the mechanical properties of density fractionated RBC were tested in this study. A non-specific NOS inhibitor, N-omega-nitro-L-arginine methyl ester (L-NAME) at a concentration of 10(-3) M and sodium nitroprusside (SNP), a nitric oxide donor at a concentration of 10(-6) M was added to blood samples with hematocrit adjusted to 0.4 l/l and RBC deformability was measured by an ektacytometer in the density fractionated RBC after one hour incubation at 37 degrees C. There was no significant effect of the NO donor SNP on cellular deformability in the older (denser) RBC fraction in contrast with the younger (least dense) fraction. Alternatively, the sensitivity of cellular deformability to competitive NOS inhibition by L-NAME was greater in the older fraction. These findings suggest that older RBC are characterized by diminished internal NO synthesis and are also less sensitive to external NO indicating that the target mechanisms for NO may also be deteriorated. PMID- 16317246 TI - Short-term isocapnic hypoxia and coagulation activation in patients with sleep apnea. AB - Hemostatic changes might contribute to the increased risk of cardiovascular and cerebrovascular events in patients with obstructive sleep apnea (OSA). We investigated the effect of a short-term isocapnic hypoxic challenge on coagulation activation markers thrombin/antithrombin III complexes (TAT) and D dimer in OSA. Thirty-two OSA patients (mean age 48 +/- 11 years) inhaled a gas mixture containing 10% O(2) and 90% N(2) and further adjusted to yield pulse oximetry saturation of 80-85% for 5 minutes. Plasma levels of TAT and D-dimer were measured immediately before and immediately after the hypoxic challenge. The hypoxic challenge provoked a significant increase in TAT (p < 0.001) and in D dimer (p = 0.037). Mean nocturnal oxygen saturation from the sleep recordings correlated with D-dimer increase (r = -0.37, p = 0.041). Also, OSA patients with a history of hypertensive parents had greater D-dimer increase in response to hypoxia than patients having normotensive parents (p = 0.035). Parental hypertension independently explained 15% of the variance in D-dimer increase after hypoxia (p = 0.035). Oxygen desaturation during sleep may predispose OSA patients, in particular those with a parental history of hypertension, to a hypercoagulable state providing one explanation for the increased risk of atherothrombotic events in this population. PMID- 16317247 TI - Comparative evaluation of two newly developed devices for capillary viscometry. AB - Viscometry is an often applied method in clinical chemistry. A variety of studies demonstrate an association of parameters related to blood viscosity with human pathology of varying origin. Whole blood and plasma viscosity are considered to be clinically useful indicators in the diagnostic workup and therapy monitoring of certain diseases. In this study, we compare the "Waegeviskosimeter" (WV) described in previous publications with a newly developed device, the "Reverse Flow Viscometer" (RFV). Both viscometers are capillary flow viscometers. Both overcome the disadvantage of common viscometers of the Ubbelohde and Cannon Fenske type which require large amounts of plasma and which can be only applied to Newtonian fluids. The accuracy of the measurements of both viscometers, requiring less than 1.0 ml sample volume, is superior to most conventional methods. The major distinction in the functionality of the WV and the RFV is that the WV measures the kinematic viscosity whereas the RFV directly estimates dynamic viscosity without the requirement of additional density measurement. We found good reproducibility of viscosity with coefficient of variation CV < or =1.1% for both viscometers. Quality assurance measures have been carried out. Because no quality assurance scheme according to the guidelines proposed by the German Medical Association exists for plasma or whole blood viscosity, we tested reference material Lyphochek Unassayed Chemistry Control Level 1 and Level 2 (Bio Rad Laboratories). We determined the viscosities 1.40 mPa s and 1.08 mPa s (37 degrees C) and the between-run precision from daily quality control runs with CV of 1.4% and 1.2% for the WV, and 1.7% and 1.4% for the RFV. For direct comparison reasons, we determined the viscosity in seventy human plasma and serum samples by both methods. Using the regression analysis described by Passing-Bablok, the RFV and the WV methods are highly correlated and show only little variations (r = 0.990, tau = 0.896). The regression equation is y(WV) = 1.035x(RFV) - 0.056 with a mean deviation of 0.4+/-3.6%. We conclude that both new devices for viscosity assessment fulfill all quality requirements as prescribed for clinical chemical laboratories. One advantage RFV is to measure the dynamic viscosity directly. PMID- 16317248 TI - Erythrocyte aggregation in Behcet's disease determined with the Sefam and Myrenne aggregometer. Lack of association with thrombosis and uveitis. AB - Behcet's disease (BD) is a chronic systemic vasculitis characterised by recurrent oral and genital ulcers and uveitis, in which 25-30% of patients develop thrombotic events of unknown etiology. In order to ascertain whether erythrocyte aggregation (EA) may be involved in thrombotic events and or uveitis in BD patients, we determined using two erythrocyte aggregometers i.e. Myrenne and Sefam (which provides the total disaggregation threshold, needed for erythrocytes to disaggregate), EA in 77 BD patients (42 male, 35 female, aged 44 +/- 12 years) and 77 controls (41 male, 36 female, aged 43 +/- 11 years). BD patients showed higher EA determined with both aggregometers: Myrenne (EA(0): P = 0.035; EA(1): P < 0.001) or the Sefam (Ta: P < 0.001, AI(10): P < 0.001, gammaD: P = 0.014) as well as higher fibrinogen and triglyceride levels (P < 0.001, P = 0.003, respectively) compared with the control group. However no differences were observed in any of the aggregation parameters determined either with the Myrenne (EA(0), EA(1)) or the Sefam (Ta, AI(10), gammaD) aggregometer when BD patients with thrombotic events (n = 23) or uveitis (n = 21) were compared with those who did not (P > 0.05). These results reinforce previous findings of our group, suggesting that EA does not seem to be involved in thrombotic events or in uveitis in BD patients. PMID- 16317249 TI - Hemodynamic alteration in diabetic nephropathy measured by renal Doppler ultrasonography: does it predict the outcome of the disease? PMID- 16317251 TI - Frontotemporal dementia: introduction. PMID- 16317252 TI - Psychiatric symptoms and their psychosocial consequences in frontotemporal dementia. AB - Based on a retrospective study of 19 neuropathologically verified cases with frontotemporal dementia (FTD), neuropsychiatric symptoms related to behavioral disturbances and their psychosocial consequences were studied. The results indicate that frontotemporal dementia is often misdiagnosed early in the clinical course. Behavioural features with impaired social interactions, impaired personal regulation, and loss of insight were seen in all patients. The psychosocial consequences reported in this paper challenge future research in frontotemporal dementia. PMID- 16317253 TI - Frontotemporal dementia: one disease, or many?: probably one, possibly two. AB - Accumulating evidence suggest that frontotemporal dementia is best viewed as a clinical syndrome even though there are distinct presentations of the behavioral variety, progressive aphasia, semantic dementia, corticobasal degeneration and progressive supranuclear palsy. Similarly the pathology should be regarded as a spectrum even though histological varieties are distinguished. More than half of FTD pathology is associated with ubiquitin positive and tau negative inclusions that are common in ALS. However the majority of FTD cases do not have ALS clinically and relatively few ALS cases develop FTD. The pathological and biochemical varieties can be dichotomized as tau positive and tau negative pathology and biochemistry. The genetics of the tau positive variety is associated with tau mutations and so far the tau negative variety is not, although some are linked to chromosome-17 also. There is a corresponding clinical dichotomy combining the behavioral variety of FTD presentation with semantic dementia and usually ubiquitin positive tau negative pathology on one hand and the association of primary progressive aphasia and cortical basal degeneration/PSP syndrome with tau positive pathology on the other. The overlap between them is too great to establish two separate diseases. PMID- 16317254 TI - The correlation of cognitive decline with frontotemporal dementia induced annualized gray matter loss using diffeomorphic morphometry. AB - This study uses large deformation medical image registration to analyze, in a disease-specific normalized space, the annual rate of gray matter atrophy caused by frontotemporal dementia (FTD) and its correlation with cognitive decline. The analysis consists of three parts. First, a labeled structural MRI atlas is deformed into the shape of an average FTD brain. Second, annualized FTD-related atrophy of gray matter structures is estimated for each patient in the database. Third, the group-wise annualized atrophy rate caused by FTD is correlated, for each gray matter voxel, with declining performance on cognitive tests. This study gives insight into the relationship between FTD-related progressive cortical atrophy and loss in cognitive function. PMID- 16317256 TI - Clinical features of frontotemporal dementia. AB - What was once called Pick's disease has three major anatomic variants. With all three, frontotemporal brain is selectively injured whereas posterior cortical regions are spared. These three clinical patterns include a bifrontal, slightly asymmetric subtype with more involvement of the right frontotemporal region called frontotemporal dementia or the frontal variant of FTD (fvFTD), a temporal predominant subtype called the temporal variant of FTD or semantic dementia (SD), and a left frontal-predominant subtype called progressive nonfluent aphasia (PNFA). The three anatomic groups help to classify distinctive clinical syndromes with unique features. Careful study of these subtypes of frontotemporal dementia, using combinations of new quantitative neuroimaging, behavioral and physiological measures are yielding important information about the functioning of the brain's frontal and temporal regions. As we come to better understand the biologic basis for the three FTD clinical syndromes, new classification schemas may emerge, but our current clinical criteria serve as a strong guide to the diagnosis and separation of FTD from Alzheimer disease and other dementias. PMID- 16317255 TI - Tau alternative splicing and frontotemporal dementia. AB - A number of neurodegenerative diseases are characterized by the presence of abundant deposits containing Tau protein. Expression of the human tau gene is under complex regulation. Mutations in the tau gene have been identified in patients with frontotemporal lobe dementia. These mutations affect either biochemical/biophysical properties or the delicate balance of different splicing isoforms. In this review, we summarize recent advances in our understanding of genetics and molecular pathogenesis of tauopathies with the focus on frontotemporal lobe dementia. We review published studies on tau pre-mRNA splicing regulation. Understanding molecular mechanisms of tauopathies may help in developing effective therapies for neurodegenerative tauopathies and related disorders, including Alzheimer disease. PMID- 16317257 TI - Ubiquitin immunohistochemistry of frontotemporal lobar degeneration differentiates cases with and without motor neuron disease. AB - Frontotemporal lobar degeneration (FTLD) without tau pathology is clinically and pathologically heterogeneous. The present report describes the neuropathology of 52 brains with FTLD without tau pathology compared with 10 brains of amyotrophic lateral sclerosis (ALS) without dementia using ubiquitin immunohistochemistry. The 52 cases were classified into 47 cases of FTLD with motor neuron disease (MND)-type inclusions but without MND (FTLD-MNI), three cases of FTLD with MND (FTLD-MND), and two cases of dementia lacking distinctive histopathology (DLDH) based on the features of ubiquitin-immunoreactive (ubiquitin-ir) structures in the caudate, frontotemporal cortices and dentate fascia, and presence or absence of neuronal loss in lower motor neurons. Many ubiquitin-ir neuronal inclusions and neurites in the caudate nucleus, frontotemporal cortices, and ubiquitin-ir crescent-or ring-shaped neuronal inclusions in the dentate fascia characterized FTLD-MNI. Ubiquitin-ir neuronal intranuclear inclusions (NII) were observed in 26 of 43 cases and associated with many neurites in the caudate nucleus as well as a familial history in most cases. A subset of cases had Pick-body-like inclusions in the dentate fascia and caudate nucleus with paucity of neuritic pathology and no NII; another had crescent-shaped inclusions in the dentate fascia and neuritic pathology with NII in the caudate. FTLD with MND was characterized by a few or no ubiquitin-ir inclusions in the caudate nucleus and frontotemporal cortices and ubiquitin-ir granular inclusions in the dentate fascia, as well as loss of lower motor neurons. These features were similar to ALS, but different from FTLD-MNI. The findings suggest that FTLD-MNI has a different pathogenesis from FTLD-MND and ALS. PMID- 16317258 TI - Autosomal dominant inclusion body myopathy, Paget disease of bone, and frontotemporal dementia. AB - Autosomal dominant proximal limb girdle or inclusion body myopathy, associated with Paget disease of bone and frontotemporal dementia (IBMPFD) is a recently described disorder that maps to chromosome 9p21.1-p12. We refined the critical locus and identified the gene as the Valosin Containing Protein (VCP) gene, a member of the AAA-ATPase superfamily using a candidate gene approach. Six missense mutations were found to co-segregate with affected individuals only, two of these representing mutation hot spots. We report the clinical and molecular findings in 99 individuals in 13 families. VCP is associated with a variety of cellular activities, including the control of cell cycle, membrane fusion, and the ubiquitin-proteasome degradation pathway. Previous studies have associated VCP mutants in cell lines with vacuole formation and aggregate formation. Identification of VCP as the gene causing IBMPFD has important implications for understanding the pathogenesis of neurodegenerative disorders. PMID- 16317259 TI - Semantic dementia and primary progressive aphasia: a problem of categorization? AB - The relationship between semantic dementia (SD) and primary progressive aphasia (PPA) has been the subject of debate ever since the syndromes were first described, in converging streams of research from the neuropsychological and neurologic communities. The most salient clinical features of SD are anomia with circumlocution and semantic paraphasia, single-word comprehension deficit, and reduced category fluency. Of critical importance is the fact that patients also show deficits on non-verbal tasks using visual, auditory, and other modalities, suggesting that the key impairment in SD is a breakdown in conceptual knowledge rather than a specific problem with language. The finding of item consistency between the various tests supports this view. The order in which the features appear can be explained by the variable degree of redundancy in access to semantic knowledge from the different perceptual modalities. Atrophy is seen in the anterior and inferior temporal lobe rather than in classic language areas, further distancing SD from aphasic syndromes. Semantic dementia and progressive non-fluent aphasia (PNFA) share some clinical and pathologic characteristics with frontal variant frontotemporal dementia, but there are also clear differences between the three syndromes. We believe that many patients described as having fluent primary progressive aphasia in fact have early SD. Semantic dementia is a well-defined syndrome, distinct from PNFA but related to it within the spectrum of frontotemporal lobar degeneration syndromes. PMID- 16317260 TI - Radiotherapy in the treatment of dermatofibrosarcoma protuberans. AB - OBJECTIVE: To evaluate the role of postoperative radiotherapy (RT) for patients with dermatofibrosarcoma protuberans (DFSP). METHODS AND MATERIALS: Ten patients were treated with surgery and postoperative RT and had follow-up from 21 to 185 months. Nine patients had DFSP and 1 patient had a fibrosarcomatous component (DFSP-FS). No patient had gross disease at the time of RT. RESULTS: Nine patients with DFSP remained disease free after surgery and postoperative RT. The patient with DFSP-FS experienced a local recurrence after surgery, external beam RT, and a brachytherapy boost, and died with disease. There were no severe complications. CONCLUSION: Postoperative RT may reduce the risk of local recurrence in patients with DFSP who have a high risk of residual disease after surgery. PMID- 16317261 TI - Pegylated liposomal doxorubicin-based combination chemotherapy as salvage treatment in patients with advanced hepatocellular carcinoma. AB - OBJECTIVES: Effective chemotherapy with the least toxicity is important for patients with inoperable or advanced hepatocellular carcinoma. Single use of pegylated liposomal doxorubicin has been reported to be safe and effective with varying treatment response. The authors evaluate its activity in combination with capecitabine or gemcitabine as salvage therapy in these patients. METHODS: At first, intravenous administration of 30 mg/m2 pegylated liposomal doxorubicin for 60 minutes on day 1, with oral capecitabine 1500 mg twice daily from day 1 to day 14 every 4 weeks (trial A) was conducted. Following unfavorable results, a second trial (trial B) was performed to subsequent patients with the same pegylated liposomal doxorubicin schedule, but in combination with 1000 mg/m2 gemcitabine over 30 minutes on day 1 and day 8, followed by a 2-week rest. RESULTS: Both trials showed no objective response, with 2 patients with stable disease in each trial. In trial A, the disease control rate of all evaluative patients (complete response + partial response + stable disease) and progression-free survival of 2 responders were 20%, 164 days and 240 days versus 22%, 75 days and 73 days in trial B. The median overall survival for all patients in trials A and B were 161 days and 84 days respectively. Generally, toxicities were well managed without toxic death. CONCLUSION: Pegylated liposomal doxorubicin-based combination chemotherapy with capecitabine or gemcitabine was not effective as salvage therapy in advanced hepatocellular carcinoma. Further effective systemic chemotherapy for patients with advanced hepatocellular carcinoma is warranted. PMID- 16317262 TI - A multicenter phase II study of "adjuvant" irinotecan following resection of colorectal hepatic metastases. AB - OBJECTIVES: This study evaluates the efficacy and toxicity of single-agent irinotecan following hepatectomy for metachronous colorectal metastases, and examines the predictive value of p27 and p53 expression and of microsatellite instability (MSI) status. METHODS: Twenty-nine patients, previously treated with 5-fluorouracil, with operable hepatic colorectal metastases underwent hepatectomy and received adjuvant irinotecan (thrice weekly) for 6 planned cycles. Metastases were examined for p53 and p27 expression by immunohistochemistry and for MSI using mono- and dinucleotide markers. RESULTS: The starting dose of irinotecan was 350 mg/m2 (in 3 patients), 300 mg/m2 (n = 14), and 250 mg/m2 (n = 12). Four patients failed to complete 6 cycles (2 progressive disease and 2 toxicity). Grade > or =3 toxicity was experienced in 8% of cycles (13 of 165). The estimated median relapse-free survival (RFS) was 45.2 months. RFS at 18 months was estimated to be 59% (95% confidence interval [CI], 43-80), 2-year overall survival (OS) was 85% (95% CI, 72-99.8), and the median follow-up was 27.9 months. Six patients (21%) have died; median OS has not been reached. In univariate analyses, p27 and MSI status were not predictive for RFS while p53 approached statistical significance (P = 0.051). Duration of chemotherapy was the only significant predictive factor (P = 0.006). CONCLUSION: The tolerability of this regimen after major liver resection supports further evaluation of irinotecan-based adjuvant chemotherapy in this group of patients. PMID- 16317263 TI - The experience of concurrent chemoradiation for Japanese patients with superficial esophageal squamous cell carcinoma: a retrospective study. AB - PURPOSE: To evaluate the treatment outcome of concurrent chemoradiation therapy (cCRT) for 13 clinically confirmed T1 esophageal cancer patients. METHODS: Between June 2000 and February 2004, patients with T1 esophageal cancer (tumor invading lamina propria or submucosa) received cCRT (50.4 Gy; CDDP: 75 mg/m2, day 1, bolus; 5-FU: 1000 mg/m2, days 1 to 4, continuous) (n = 13, T1 group). This treatment regimen was compared with the following 2 other groups treated during the same period: one was treated with radiation therapy alone for T1 disease (n = 5, RT-alone group); and the other group of patients consisted of those with T2 disease (tumor invading muscularis propria) who received the same cCRT regimen as the first T1 group (n = 9, T2 group). RESULTS: The overall survival rates at 1 and 3 years were in the T1 group, 75% and 75%; in the T2 group, 75% and 45% (P =0.2890); and in the RT-alone group, 60% and 40% (P = 0.2978). No treatment related mortalities occurred in the T1 and RT-alone groups, but one patient in the T2 group died of radiation-induced pneumonitis. CONCLUSIONS: Although this study was not randomized, the results showed that cCRT is a safe and effective method for treating patients with superficial esophageal cancer. PMID- 16317264 TI - Cyclooxygenase-2 (COX-2) levels before and after chemotherapy: a study in rectal cancer. AB - OBJECTIVES: Induction of cyclooxygenase-2 (COX-2) by inflammatory mediators, oncogenes, and carcinogens has been demonstrated in preclinical models. However, there are limited clinical data regarding COX-2 induction by chemotherapy or radiation. Experimental data suggest cross-talk between the EGFR and COX-2 pathways. The aim of this study was to analyze the expression of COX-2 before and after chemoradiation (CRT) and correlate the same with tumor (T) down-staging and survival. Similar data were obtained for EGFR expression before and after chemoradiation. METHODS: Archival paraffin-embedded tumor specimens from patients undergoing CRT between 1995 and 2001 were analyzed. COX-2 expression was measured by immunohistochemistry (IHC), using the 160112 COX-2 mouse monoclonal antibody. For EGFR, we used mouse monoclonal Ab-10. Standard immunoperoxidase technique was used to detect the avidin- biotin peroxidase complex. Staining in tumor tissue was visually scored and confirmed by an image analyzer (ACIS; ChromaVision Medical Systems, Inc, San Juan Capistrano, CA). RESULTS: Twenty pretreatment biopsy samples from rectal cancer patients and their paired, post-CRT surgical specimens (n = 17) were analyzed. Three cases had no primary tumor after CRT. COX 2 expression was noted in 19 of 20 pretreatment samples and 17 of 17 surgical specimens. EGFR expression was noted in 10 cases pretreatment. Six patients with weakly positive COX-2 expression pretreatment had increased COX-2 expression after CRT, whereas in 1 patient the expression decreased after CRT. No EGFR induction was noted. There was no statistical association between EGFR and COX-2 expression in this data set. Median survival for the entire cohort was 38.9 months. There was no difference in survival between the COX-2 induced and noninduced groups. CONCLUSIONS: COX-2 induction was seen with CRT in this population of rectal cancer patients. Prognostic significance of this induction remains to be defined in a larger cohort. PMID- 16317265 TI - Efficacy and tolerability of oxaliplatin plus irinotecan 5-fluouracil and leucovorin regimen in advanced stage colorectal cancer patients pretreated with irinotecan 5-fluouracil and leucovorin. AB - OBJECTIVES: Oxaliplatin has been introduced in the treatment of advanced colorectal cancer during the past few years. The pre-existing treatment of leucovorin-5-fluorouracil-irinotecan (IFL), although reasonably effective, has needed novel, active agents to increase the response rate and overall survival. We planned this phase 2 study in patients pretreated with IFL, adding oxaliplatin as second-line treatment: our objectives were to determine response rate and overall survival. METHODS: All patients (median age 65) were designated to receive 6 cycles of chemotherapy: leucovorin 200 mg/m2 infused for 60 minutes, 5 fluorouracil 500 mg/m2 bolus at 30 minutes from the start of the previous infusion, irinotecan 135 mg/m2 infused for 90 minutes, and oxaliplatin 135 mg/m2 for 90 minutes, infused sequentially on day 1 and repeated every 3 weeks. Standard ondansetron antiemetic treatment and dexamethasone 8 mg were administered to all patients. No prophylactic recombinant human granulocyte colony-stimulating factor was permitted. RESULTS: Fifty-seven patients were recruited and 54 were evaluable for response, survival, and toxicity. All patients had advanced, inoperable, metastatic disease in the liver and/or lungs, abdominal cavity, and multiple sites. All patients had undergone IFL pretreatment and had no response; 40 had disease progression and 14 had stable disease when entering the present study; 302 chemotherapy cycles (mean 5.92) were administered. There was no treatment delay caused by toxicity (either neutropenia or diarrhea). Irinotecan and oxaliplatin were reduced by 25% in 6 (11.1%) patients. No complete responses were observed; 21 (38.9%) patients achieved partial response, 26 (48.2%) had stable disease, and 7 (13%) had disease progression. Median duration of response was 6 months, time to tumor progression (TTP) 8 months, and median overall survival after the initiation of second-line treatment was 10 months (95% confidence interval [CI], 7.5-12.6). CONCLUSION: The addition of oxaliplatin to IFL as second-line treatment rendered a prolongation of survival and a response rate of 38.9% in patients in whom IFL pretreatment had failed. PMID- 16317266 TI - Erlotinib and chemoradiation followed by maintenance erlotinib for locally advanced pancreatic cancer: a phase I study. AB - OBJECTIVES: A phase I trial was conducted to determine the maximally tolerated dose of erlotinib with concurrent gemcitabine, paclitaxel, and radiation for patients with locally advanced pancreatic cancer and to gather preliminary data on maintenance erlotinib after chemoradiation. METHODS: Patients received gemcitabine, 75 mg/m2, and paclitaxel, 40 mg/m, weekly for 6 weeks with 50.4 radiation to the primary tumor and draining lymph nodes with a 2- to 3-cm margin. Erlotinib was administered over 3-dose levels (50-100 mg/d) with chemoradiation then all patients received 150 mg/d maintenance until disease progression. RESULTS: Seventeen patients were assessable for toxicity; 13 with locally advanced disease and 4 who had undergone resection but had positive margins. At erlotinib dosages > or =75 mg/d with chemoradiation the dose-limiting toxicities were diarrhea, dehydration, rash, myelosuppression, and small bowel stricture. Maintenance erlotinib, 150 mg/d, was well tolerated. The median survival of the 13 patients with locally advanced disease was 14.0 months and 6 of 13 (46%) had a partial response. CONCLUSIONS: The maximum tolerated dose of erlotinib with gemcitabine, paclitaxel and concurrent radiation is 50 mg/d for patients with locally advanced pancreatic cancer. Full dose maintenance erlotinib is well tolerated. Promising preliminary activity and overall survival were demonstrated. PMID- 16317267 TI - Retrospective clinicopathologic correlation of gross tumor size of hepatocellular carcinoma: implications for stereotactic body radiotherapy. AB - PURPOSE: To determine the degree of correlation between radiographic size and true gross pathologic size for subjects with primary hepatocellular carcinoma (HCC). METHODS AND MATERIALS: This analysis included 18 patients with 27 tumors who underwent either partial hepatectomy or orthotopic liver transplantation for HCC at the University of Colorado Hospital between 1997 and 2002. Preoperative imaging was performed using computed tomography (CT) or magnetic resonance imaging (MRI). After surgical resection the size of each tumor on gross pathologic examination was recorded. The maximal measurement in one dimension on axial imaging and pathologic examination was extracted for statistical analysis. The clinical and pathologic sizes were compared using a percent size difference (%Deltasize) as an end point for each patient. A regression analysis was applied to study the association between pathologic and clinical size. RESULTS: The median radiographic size was 2.90 cm (range 1.2-4.9). The median pathologic size was 2.50 cm (range 1-4.8). The radiographic size was larger than or equal to the pathologic size in 22/27 tumors (81%) and smaller in 5/27 (19%) tumors. The median %Deltasize was 17.5% (range -20-144%). Overall, the radiographic and pathologic sizes were positively correlated (r = 0.8). This correlation was not affected by choice of imaging modality (CT versus MRI, P = 0.71) or time of preoperative imaging (0-4 weeks versus 4-8 weeks before surgery, P = 0.61). CONCLUSIONS: Our study shows that in most instances (81%), imaging by CT or MRI overestimates true gross pathologic size of HCC. Nineteen percent of tumors appeared smaller on preoperative imaging than on the final pathologic specimen. Radiation therapy utilizing a 0.5 or 1.0 cm margin around the radiographic tumor would have encompassed the gross pathologic tumor in 93% and 100% of cases, respectively. PMID- 16317268 TI - Irinotecan (CPT-11) and mitomycin-C (MMC) as second-line therapy in advanced gastric cancer: a phase II study of the Gruppo Oncologico dell' Italia Meridionale (prot. 2106). AB - OBJECTIVE: The aim of this study was to evaluate the activity and toxicity of a combination regimen of CPT-11 and mitomycin-c as second-line chemotherapy for pretreated patients with advanced, metastatic, or both, gastric adenocarcinoma. MATERIALS AND METHODS: Patients with pretreated metastatic disease or early relapsed after adjuvant chemotherapy were enrolled. Entry criteria included histologic/cytologic diagnosis of gastric adenocarcinoma, age 18 to 75 years, performance status > or =70 (Karnofsky scale), bi-dimensionally measurable disease. Patients received CPT-11 and mitomycin-c at the dosage of 150 mg/m2 on days 1 and 15, and 8 mg/m2 on day 1, respectively, every 4 weeks. The disease evaluation was done every 3 cycles. RESULTS: Among the 38 patients we observed, 1 (3%) complete response and 11 (29%) partial responses for an overall response rate of 32% according to an intent-to-treat analysis. The median duration of response was 6.5 months. The median time to progression was 4 months with a median overall survival 8 months. All patients were evaluable for toxicity and the only grade 3-4 observed toxicities were leukopenia (8%), neutropenia (21%), and anemia (5%). CONCLUSIONS: The combination of CPT-11 and mitomycin-c is an active and well tolerated second-line treatment in pretreated gastric cancer patients. Further studies are needed to test its role in first-line treatment. PMID- 16317269 TI - Neuron-specific enolase and lung cancer. AB - OBJECTIVE: Serum and bronchoalveolar lavage fluid (BALF) neuron-specific enolase (NSE) levels in lung cancer have been investigated widely; however, their diagnostic values have not yet been clarified. The authors investigated the diagnostic validity of NSE in BALF and serum in lung cancer. MATERIALS AND METHODS: In this prospective case-control study, NSE levels in BALF (B-NSE) and serum (S-NSE) of 3 groups of subjects were analyzed: control subjects (group 1, n = 15), patients with chronic obstructive pulmonary disease (COPD; group 2, n = 15), and lung cancer (group 3, n = 35). RESULTS: The differences in S-NSE and B NSE levels between the groups were not significant (P > 0.05). S-NSE and B-NSE levels did not show any difference between smokers and nonsmokers, small cell lung cancer and nonsmall cell lung cancer patients, and stage I-II and stage III IV patients in group 3 (P > 0.05). B-NSE or B-NSE/urea did not show any significance in comparison with S-NSE in the diagnosis and/or staging of malignancy (P > 0.05). S-NSE and B-NSE were well correlated with each other (r = 0.84, P = 0.000). The sensitivity of the S-NSE was 60% and the specificity was 40%. CONCLUSION: The authors conclude that, although elevation of B-NSE is a well known parameter in small cell lung cancer, it can also be elevated considerably in nonsmall cell lung cancer and COPD. Because of the significant correlation between S-NSE and B-NSE, it may be sufficient to measure S-NSE activity because it is easier and less invasive. However, NSE has no role in the exact diagnosis of lung cancer; it can only be investigated in a scientific setting. PMID- 16317270 TI - Stereotactic body radiation therapy of lung tumors: preliminary experience using normal tissue complication probability-based dose limits. AB - OBJECTIVES: To assess the feasibility and toxicity of stereotactic body radiotherapy (SBRT) for patients with locally advanced or metastatic tumors in lung. METHODS: Twenty-five tumors in 17 patients were treated. All treatments were delivered in 3 daily fractions of 9 to 15 Gy per fraction. Normal tissue complication probability (NTCP) calculations (using the Lyman model) were performed to facilitate dose prescription, and doses were prescribed with a maximum allowable NTCP risk of pneumonitis of up to 20%, not to exceed 15 Gy per fraction. Planning target volumes were designed to allow for respiratory variation in tumor location. RESULTS: The median dose prescribed was 35 Gy (range, 24 to 45 Gy). Twenty-three of 25 tumors remained controlled at median follow-up of 14 months. Four patients experienced grade 1-2 acute toxicity. Late toxicity developed in 2 patients who received treatment to peri-hilar tumors, including one patient in whom bronchial stenosis developed with complete occlusion and lobar atelectasis 6 months after treatment. No patient had grade 3 or 4 radiation pneumonitis. CONCLUSIONS: SBRT prescribed within the confines of NTCP-restricted dosing on this protocol resulted in no radiation pneumonitis. Tissues other than lung parenchyma which are unaccounted for by NTCP may be dose limiting when performing hypofractionated SBRT in the lung. PMID- 16317271 TI - Re-examining the role of elective nodal irradiation: finding ways to maximize the therapeutic ratio. AB - OBJECTIVE: Elective nodal irradiation (ENI) of regional lymphatics has been a foundational paradigm for radiation oncologists in the treatment of nonsmall-cell lung cancer (NSCLC), but its utility has recently been called into question. This review summarizes the controversies surrounding ENI and reviews the therapeutic options available to treat regional lymphatics in NSCLC. METHODS: Local failure after conventional radiotherapy (RT) occurs in 40% to 80% of patients fueling the investigation of more aggressive RT regimens. As the dose is increased and accelerated the volume of normal lung tissue treated becomes a limiting factor. Thus elimination of ENI followed by further dose escalation has become a commonly pursued solution. When ENI is excluded, treatment is restricted to clinically positive disease and negative lymph node stations are left untreated. RESULTS: Radiographic and surgical data suggest our ability to determine the true extent of disease is imperfect and therefore the elimination of ENI likely leaves microscopic NSCLC untreated. CONCLUSIONS: At our institution we have concluded that the prophylactic treatment of regional lymph nodes is best reserved for patients most likely to achieve local control and are designing treatment protocols including chemotherapy to take advantage of this improvement in local control. PMID- 16317272 TI - Cataracts among cancer survivors. AB - OBJECTIVES: To determine the frequency of cataracts in cancer survivors and their association with patients' demographic characteristics, treatment, and cancer type and the perceived effect cancer had had on their overall health. METHODS: This was a descriptive, cross-sectional review of the results of a survey of long term cancer survivors who had been treated at the authors' institution. RESULTS: Of 3936 respondents to the survey, 168 (4%) reported having had cataracts. Cataracts were most frequent in survivors of hematologic diseases (chronic leukemia, 17%; myeloma, 13%; acute leukemia, 9%; and lymphoma, 7%). There were no notable associations between cataracts and gender, age, ethnicity, marital status, education completed, or work status. Cancer survivors with cataracts were more likely to report that cancer had affected their overall health (56% vs 39%). There was a trend of association between cataracts and prior treatment with bone marrow transplantation, but not corticosteroids. CONCLUSION: Cataracts affect a minority of long-term cancer survivors, but those who reported them were more likely to report that cancer had had a lasting effect on their overall health. Vision evaluation for all cancer survivors is recommended. PMID- 16317273 TI - Prospective trial of a video educational tool for radiation oncology patients. AB - OBJECTIVES: Prospective assessment of the informational needs of radiation oncology patients and efficacy of an educational video in meeting them. METHODS: Subjects completed baseline self-administered questionnaires and subsequently viewed the patient education video. Post-testing was performed after initiation of therapy and subjects rated their satisfaction with the video, its relevance, and their emotional response. Analyses were performed with respect to patient and disease characteristics. RESULTS: Fifty-three subjects were enrolled and completed both before and after video measures. The mean age of participants was 58 years (range, 33 to 83). Pretreatment, >90% of patients reported specific information needs. One hundred percent of patients watched the video and 77% rated it as highly relevant. High levels of satisfaction (>90%) were reported with video information describing radiation and simulation. Older subjects (58 years and older) found video information significantly more relevant than those younger (55% versus 27%, P = 0.04) and rated greater satisfaction with side effect information (78% versus 41%, P = 0.006). Subjects with breast cancer exhibited a trend towards feeling better informed by the video. CONCLUSIONS: Radiation oncology patients reported informational needs unmet by standard educational measures. High levels of satisfaction were reported with video education. It promoted better understanding of radiotherapy. Older patients found the video to be significantly more relevant and informative. PMID- 16317274 TI - Phase I study of piritrexim and gemcitabine in patients with advanced solid tumors. AB - OBJECTIVES: In this phase I study, the combination of piritrexim and gemcitabine was given to establish the maximum tolerated dose and the recommended phase II dose, and to determine a toxicity and efficacy profile. METHODS: Fifty-two patients with normal and impaired renal function were enrolled on this phase I study. The starting dose was piritrexim 10 mg 3 times daily (5 days of the week for 3 weeks and 1 week off each 28-day cycle) and gemcitabine 1000 mg/m2 on days 1, 8, and 15. The piritrexim was escalated in a stepwise fashion with this dose of gemcitabine and then with gemcitabine 1000 mg/m2 for days 1 and 15. RESULTS: The recommended phase II dose of this combination was felt to be piritrexim 50 mg/day (10 mg every morning, 20 mg every noon, and 20 mg every evening) with gemcitabine 1000 mg/m2 on days 1, 8, and 15, and piritrexim 75 mg/day (25 mg thrice daily) with gemcitabine 1000 mg/m2 on days 1 and 15. Neutropenia and thrombocytopenia were the most often reported toxicity. Dose-limiting toxicity was thrombocytopenia in both groups. The number of renal-impaired patients enrolled was too small to establish a maximum tolerated dose for this group (piritrexim became unavailable), but the combination was tolerated in the patients with impaired renal dysfunction. There was 1 complete response, 1 partial response, and 1 minimal response. CONCLUSION: The combination of piritrexim and gemcitabine was determined to be tolerable in heavily pretreated patients for use in solid tumors. PMID- 16317275 TI - Comparison of treatment volumes and techniques in prostate cancer radiation therapy. AB - OBJECTIVE: To compare dose-volume histograms (DVHs) for 3 target volumes (group 1, prostate + seminal vesicles + pelvic lymph nodes; group 2, prostate + seminal vesicles; group 3, prostate only) to determine the difference in dose to normal structures (rectum, bladder, and femoral heads) while controlling for target dose using 3-dimensional conformal radiation therapy (3DCRT) versus intensity modulated radiation therapy (IMRT). METHODS: Ten patients with localized prostate cancer were randomly selected. 3DCRT and IMRT planning were done to deliver 75.6 Gy to the prostate, 50.4 Gy to the pelvic nodes, and 55.8 Gy to the seminal vesicles at a standard fractionation of 1.8 Gy. An additional plan delivering 75.6 Gy to the seminal vesicles using IMRT was run. DVHs were compared for 3DCRT and IMRT. RESULTS: In all 3 groups, the percent rectum receiving > or =70 Gy, > or =60 Gy, and > or =40 Gy was significantly less for IMRT than for 3DCRT. Increasing target volumes, as necessary for pelvic nodal irradiation, overall did not result in higher rectal doses for IMRT. With 3DCRT, however, larger target volumes did increase the amount of rectum irradiated. Similar results were obtained for the femoral heads whereas results for the bladder were mixed. CONCLUSION: When compared with 3DCRT, IMRT delivered equivalent or higher doses to the target volume with greater sparing of critical organs. Because dose-volume parameters have been shown to relate to toxicity, IMRT would appear to be the favored technique for prostate cancer radiation, particularly with regard to nodal treatment. PMID- 16317276 TI - Head and neck mucosal melanoma. AB - PURPOSE: The purpose of this article is to discuss the optimal treatment and outcomes for head and neck mucosal melanoma. METHODS: Review the pertinent literature. RESULTS: Head and neck mucosal melanoma is a rare entity comprising less than 1% for all Western melanomas. It usually arises in the nasal cavity, paranasal sinuses, and oral cavity. The optimal treatment is surgery. The likelihood of local recurrence after resection is approximately 50%. Radiotherapy (RT) reduces the likelihood of local failure but probably does not enhance survival, which is primarily impacted by advanced T stage and the presence of regional metastases. The 5-year survival rates vary from approximately 20 to 50%. Although the median time to relapse is roughly 1 year or less, late failures are common and cause-specific survival continues to decline after 5 years. CONCLUSION: The optimal treatment is surgery. Postoperative RT improves local regional control but may not impact survival. Definitive RT may occasionally cure patients with unresectable local-regional disease or at least provide long-term palliation. PMID- 16317277 TI - Multitargeted therapy in estrogen receptor-positive, human epidermal growth factor receptor-2-positive breast cancer. PMID- 16317278 TI - Re: The effects of ondansetron and granisetron on electrocardiography in children receiving chemotherapy for acute leukemia. PMID- 16317279 TI - Adjuvant chemotherapy for Merkel cell carcinoma. PMID- 16317280 TI - Presentation of esophageal cancer with solitary splenic metastasis. PMID- 16317281 TI - Transarterial chemoembolization in a patient with recurrent hepatocellular carcinoma and portal vein thrombosis: a case report and review of the literature. AB - Primary hepatocellular carcinoma (HCC) accounts for more than 5% of all cancers in the world, and causes between 250,000 and one million deaths globally each year. The incidence of portal vein thrombosis in HCC patients is around 8%. These patients are regarded as unsuitable for chemoembolization, especially if there is portal vein occlusion. The authors describe a patient with HCC and partial portal vein thrombosis who successfully underwent transarterial chemoembolization. PMID- 16317282 TI - Benign pleural schwannoma secondary to radiotherapy for Hodgkin disease. PMID- 16317283 TI - Thyroid metastasis of malignant melanoma. AB - Clinically significant metastases to the thyroid gland are very rare; however, they can present as the initial malignancy. The authors report a 53-year-old man who underwent surgery for malignant melanoma 5 years earlier, and recently presented with a thyroid nodule that turned out to be a metastasis of the primary malignancy. Fine-needle aspiration showed malignant cells, but was unable to indicate that their origin was malignant melanoma. Total thyroidectomy was the therapy chosen for the patient. PMID- 16317284 TI - Interleukin-2 for the treatment of solid tumors other than melanoma and renal cell carcinoma. AB - Interleukin-2 (IL-2) is a lymphokine produced by T cells whose main function is to stimulate the growth and cytotoxic response of activated T lymphocytes. It has been used to stimulate the immune system for the treatment of multiples tumors. This article is intended to review the reports published from 1990 to 2004 on the IL-2 treatment of tumors other than melanoma and renal carcinoma. A literature search was made in various databases (MEDLINE, EMBASE and BioAssay), focused on IL-2 clinical efficacy in such tumors. A selection was made over 150 publications reporting on administration of IL-2 in multiple tumors: lung carcinoma (small cell and non-small cell), colorectal, gastric, pancreatic, ovarian and breast cancer, sarcomas, hepatocarcinoma, mesothelioma, and brain, urological, and head and neck tumors. IL-2 was mainly used in metastatic disease, associated with other immunotherapy or chemotherapy schedules. We conclude that adjuvant IL-2 may be of value in early stages combined with standard treatment for colon and pancreas cancers. In other neoplasms, the indication for adjuvant IL-2 has been sporadic and does not allow conclusions to be drawn. Assessment of the efficacy of IL-2 combined with chemotherapy as treatment for advanced stages is complex, due to the lack of a control, and the variety of dosages and schemes. The activity of IL-2 in monotherapy or in association with immunotherapy is clinically relevant in hepatocarcinoma, mesothelioma and in malignant overflows as palliative treatment. Randomized trials would be required in order to be able to draw conclusions about its indication in other tumors. PMID- 16317285 TI - Molecular targeting of growth factor receptor-bound 2 (Grb2) as an anti-cancer strategy. AB - Growth factor receptor-bound 2 (Grb2) is a ubiquitously expressed adapter protein that provides a critical link between cell surface growth factor receptors and the Ras signaling pathway. As such, it has been implicated in the oncogenesis of several important human malignancies. In addition to this function, research over the last decade has revealed other fundamental roles for Grb2 in cell motility and angiogenesis--processes that also contribute to tumor growth, invasiveness and metastasis. This functional profile makes Grb2 a high priority target for anti-cancer drug development. Knowledge of Grb2 protein structure, its component Src homology domains and their respective structure-function relationships has facilitated the rapid development of sophisticated drug candidates that can penetrate cells, bind Grb2 with high affinity and potently antagonize Grb2 signaling. These novel compounds offer considerable promise in our growing arsenal of rationally designed anti-cancer therapeutics. PMID- 16317286 TI - Hypoxia-targeted bioreductive tyrosine kinase inhibitors with glutathione depleting function. AB - Tyrosine kinase inhibitors may serve as ligands for kinases that are involved in normal cell differentiation or repair, thereby leading to toxicity. It may be possible to target such inhibitors to tumor cells by coupling them to hypoxia activated bioreductive molecules. Such coupling can utilize or incorporate bonds that have a propensity to be preferentially oxidized by thiols such as intracellular glutathione (GSH). The resulting depletion of GSH may increase redox-mediated apoptosis. The resultant molecule is hence projected to act via multiple cell killing mechanisms: (i) inhibition of tumor kinases, (ii) tumor DNA disruption and (iii) causing increased redox-mediated apoptosis. PMID- 16317287 TI - NPI-2358 is a tubulin-depolymerizing agent: in-vitro evidence for activity as a tumor vascular-disrupting agent. AB - The diketopiperazine NPI-2358 is a synthetic analog of NPI-2350, a natural product isolated from Aspergillus sp., which depolymerizes microtubules in A549 human lung carcinoma cells. Although structurally different from the colchicine binding site agents reported to date, NPI-2358 binds to the colchicine-binding site of tubulin. NPI-2358 has potent in-vitro anti-tumor activity against various human tumor cell lines and maintains activity against tumor cell lines with various multidrug-resistant (MDR) profiles. In addition, when evaluated in proliferating human umbilical vein endothelial cells (HUVECs), concentrations as low as 10 nmol/l NPI-2358 induced tubulin depolymerization within 30 min. Furthermore, NPI-2358 dose dependently increases HUVEC monolayer permeability--an in-vitro model of tumor vascular collapse. NPI-2358 was compared with three tubulin-depolymerizing agents with vascular-disrupting activity: colchicine, vincristine and combretastatin A-4 (CA4). Results showed that the activity of NPI 2358 in HUVECs was more potent than either colchicine or vincristine; the profile of CA4 approached that of NPI-2358. Altogether, our data show that NPI-2358 is a potent anti-tumor agent which is active in MDR tumor cell lines, and is able to rapidly induce tubulin depolymerization and monolayer permeability in HUVECs. These data warrant further evaluation of NPI-2358 as a vascular-disrupting agent in vivo. Currently, NPI-2358 is in preclinical development for the treatment of cancer. PMID- 16317288 TI - Farnesylthiosalicylic acid: inhibition of proliferation and enhancement of apoptosis of hormone-dependent breast cancer cells. AB - Farnesyltransferase inhibitors (FTIs) are being developed to block Ras-mediated actions, but current data suggest that the FTIs act through other non-Ras pathways. A new agent, farnesylthiosalicylic acid (FTS), blocks the binding of Ras to membrane acceptor sites and causes a marked reduction in Ras levels. Accordingly, FTS could be a useful new agent for the treatment of hormone dependent breast cancer. We examined the dose-response effects of FTS on the growth of MCF-7 breast cancer cells in vitro and in vivo. Further, we dissected out its specific effects on cell proliferation and apoptosis by measuring BrdU incorporation into DNA and by using an ELISA assay to quantitate the magnitude of apoptosis. FTS and its solubilized conjoiner FTS-cyclodextrin markedly inhibited cell growth in MCF-7 breast cancer cells in culture and in xenografts. This agent exerted dual effects to reduce cell proliferation as assessed by BrdU incorporation and to enhance apoptosis as quantitated by ELISA assay. These data suggest that FTS is a promising agent to be developed for treatment of hormone dependent breast cancer. PMID- 16317289 TI - Correlations between the activities of 19 anti-tumor agents and the intracellular glutathione concentrations in a panel of 14 human cancer cell lines: comparisons with the National Cancer Institute data. AB - The aim of this work was 2-fold: (i) to identify correlations between the activities of pairs of 19 anti-tumor agents in a mini-panel of 14 human cancer cell lines of diverse origins with the goal of validating the panel, and (ii) to look for correlations between the activities of 19 standard anti-tumor agents and the intracellular concentrations of glutathione (GSH). Validation with analogous data from the National Cancer Institute (NCI) Developmental Therapeutics Program was made. The cell growth inhibition potencies of the anti-tumor agents [cisplatin, carboplatin, oxaliplatin, DACH-Pt, melphalan, chlorambucil, thiotepa, busulfan, doxorubicin, etoposide, camptothecin, vinblastine, podophyllotoxin, colchicine, taxol, hydroxyurea, methotrexate, 5-azacytidine and 5-fluorouracil (5 FU)] were estimated in 14 cancer cell lines by their GI50 values. An enzymatic assay based on the method of Tietze was employed to measure intracellular total GSH concentrations. The Delta method was used to compare pairs of anti-tumor agents; similarities and differences in activity profiles (mean graphs) were evaluated by regression analysis. Most, but not all, of the correlations could be explained based on similarities in the mechanisms of action and many correlations/non-correlations were also observed in the NCI data. Some correlations were unexpected however, and not seen in the NCI data. For example, strong positive correlations (P < 0.01) were found between the GI50 values of melphalan/chlorambucil and the anti-mitotic agents. Similarly unexpected, a strong positive correlation was observed between methotrexate and cisplatin (P < 0.01). Interestingly, moderate to strong negative correlations (P < 0.01-0.05) were found between the GI50 values of 5-FU and the anti-mitotic agents/melphalan/chlorambucil. Significant positive correlations between intracellular GSH concentrations and GI50 values were found only for thiotepa (P < 0.05) and doxorubicin (P < 0.01). Data from a NCI panel of 34 cancer cell lines showed no correlations between GSH levels and the GI50 values of the same 19 compounds. In conclusion, a panel of 14 human cancer cell lines of diverse origin was used to identify similarities and differences in the activities of standard anti-tumor agents. The level of significance was stronger with the 34 cell lines of the NCI, however. Our results indicate that GSH intracellular concentrations correlate with resistance only with doxorubicin and thiotepa in these cell lines. PMID- 16317290 TI - In-vitro and in-vivo anti-cancer activity of a novel gemcitabine-cardiolipin conjugate. AB - Our objectives were to study the biological activity of a novel gemcitabine cardiolipin conjugate (NEO6002) and compare that with gemcitabine. Cytotoxicity in vitro was determined against several gemcitabine-sensitive parental and gemcitabine-resistant cancer cell lines using the sulforhodamine B assay. The in vivo toxicity was examined by changes in body weight and hematologic indices of conventional mice. Immunodeficient SCID mice bearing P388 and BxPC-3 tumor xenografts were used to evaluate the in-vivo therapeutic efficacy. Both NEO6002 and gemcitabine showed pro-apoptotic and cytotoxic effects against all gemcitabine-sensitive cell lines tested. Unlike gemcitabine, the cytotoxicity of NEO6002 was independent of nucleoside transporter (NT) inhibitors, indicating a different internalization route of NEO6002. The conjugate demonstrated a favorable activity not only in ARAC-8C, a NT-deficient gemcitabine-resistant human leukemia cell line, but also in several other gemcitabine-resistant cell lines. At the in-vivo level, a comparative toxicity study showed a significant body weight loss and a decrease in white blood cell counts in gemcitabine-treated mice, whereas the influence of NEO6002 was mild. Treatment of NEO6002 at 27 micromol/kg increased the median survival of CD2F1 mice bearing P388 cells by up to 73%, while at the same doses and schedule of gemcitabine resulted in toxic deaths of all treated mice. At a dose of 18 micromol/kg, NEO6002 inhibited the growth of BxPC-3 xenografts by 52%, while only 32% of tumor inhibition was achieved with gemcitabine. We conclude that NEO6002 may be an effective chemotherapeutic agent with improved tolerability and can potentially circumvent NT-deficient, gemcitabine-resistant tumors. PMID- 16317291 TI - Quantification of [3H]docetaxel in feces and urine: development and validation of a combustion method. AB - Most radiolabeled biological samples require extensive sample preparation to reduce quenching interference before quantification of radioactivity is possible. Clearly, a more rapid and simple method ensuring a constant count rate and optimal counting efficiency has important advantages. We report on the development and analytical method validation of a rapid and simple combustion method to quantify [3H]docetaxel excreted in human feces and urine. A 3-day validation procedure was performed; quality control (QC) samples, prepared in blank feces and urine, were combusted 5 times and aliquots of the produced tritiated combustion water were counted in a liquid scintillation counter. The validation runs demonstrated adequate precision (below 7.6%) across all QC levels. Sensitivity at the lowest QC level was excellent and recovery of radioactivity constant (ranging from 85 to 91.8%). Clinical applicability of the method was tested in a cancer patient receiving docetaxel and a tracer amount of [3H]docetaxel; during the first 72 h after [3H]docetaxel infusion, 60% of total radioactivity was excreted in the collected feces and urine, which is within the expected range. Combustion of tritiated feces and urine samples is a simple, rapid, sensitive, precise and reproducible method with high recovery. It can be applied to quantify [3H]docetaxel excretion after i.v. administration. PMID- 16317292 TI - Effects of idarubicin and idarubicinol on rat coronary resistance and vasoconstrictor responsiveness of isolated aorta and mesentery. AB - It has been hypothesized that coronary vasoconstriction is involved in the cardiotoxic action of anthracyclines. The purpose of this study was to determine whether an increase in coronary resistance induced by idarubicin (IDA) or its primary circulating metabolite idarubicinol (IDOL) is correlated with a decrease in vascular sensitivity to vasoconstrictor agonists. Coronary vasoconstriction was studied in single-pass perfused rat hearts after a 10-min infusion of 0.5 mg IDA or IDOL. In the endothelium-intact rat thoracic aorta and mesentery we measured the inhibition of phenylephrine (PE)- and KCl-induced contraction in the presence of IDA and IDOL, respectively. The increase in coronary vascular resistance evoked by IDOL (121%) exceeded that of IDA (75%). IDA (10-100 micromol/l) concentration-dependently diminished vascular sensitivity to PE and KCl due to a reduction in maximal contractile response (Emax), i.e. the antagonism by IDA of PE- or KCl-induced vasoconstriction was non-competitive, indicating a post-receptor cellular mechanism. These reductions of PE or KCl efficacy elicited by IDOL were significantly larger than those elicited by the corresponding doses of IDA. The decrease in efficacy of PE in the presence of IDA and IDOL was characterized by IC50 estimates of 44.3 and 30.7 mumol/l, respectively. With a 10-fold lower IC50, IDA inhibited the reactivity of small mesenteric arteries to noradrenaline with 10-fold higher potency. The correlation between the increase in coronary resistance and the decrease in vasoconstrictor responsiveness may suggest that these anthracyclines act through a common cellular mechanism. PMID- 16317293 TI - Plasma kinetics of procarbazine and azo-procarbazine in humans. AB - The plasma kinetics of procarbazine (PCB) and its major metabolite azo procarbazine (azo-PCB) were systematically investigated in humans for the first time. Eight therapy-refractory tumor patients with normal liver and renal function were given a single oral dose of 300 mg PCB hydrochloride as a drinking solution under fasting conditions. With the exception of the single i.v. administration of 10 mg ondansetron hydrochloride immediately before the administration of PCB, the patients were free of any co-medication 4 weeks before and during the study. PCB and azo-PCB were determined by a specially developed HPLC-UV method. PCB was absorbed very rapidly. Mean maximum plasma concentration was 12.5 min. A high elimination rate of PCB from plasma was found. The mean apparent oral systemic clearance and the plasma elimination half-life were estimated at 35.8 l/min and 9.2 min, respectively. Considerable amounts of azo PCB are found in the plasma of the eight tumor patients. The mean Cmax and AUC ratios of azo-PCB/PCB were estimated at 5.5 and 45.2. Azo-PCB is formed very rapidly from PCB, but eliminated much more slowly from plasma than PCB. Considerable interindividual differences in the conversion rate of azo-PCB to its further metabolites were observed which should have consequences for the individual tumor therapeutic efficiency of PCB. No toxic side-effects or symptoms such as nausea or vomiting were observed during the entire study. PMID- 16317294 TI - Phase I trial of gemcitabine, doxorubicin and cisplatin (GAP) in patients with advanced solid tumors. AB - A phase I study was conducted to determine the recommended phase II dose, safety profile and anti-tumor activity of a combination regimen of gemcitabine, doxorubicin and cisplatin (GAP). Gemcitabine (G) and doxorubicin (A) were administered on days 1 and 8 at increasing doses (starting level 800 and 15 mg/m, respectively). Cisplatin (P) was given at a fixed dose of 50 mg/m2 (day 1). Treatment cycles were repeated every 3 weeks. Nineteen patients received 76 cycles of treatment. A and G were escalated up to 20 and 1000 mg/m2, and finally de-escalated to 15 and 800 mg/m2. The dose-limiting toxicity was neutropenic fever that was observed in 21% of the patients. Non-hematological toxicities included mild/moderate nausea, vomiting, diarrhea and fatigue, observed in 58, 37, 21 and 95% of the patients, respectively. Of 19 patients with evaluable disease, six patients had a partial response yielding an overall response rate of 31.6 % (95% confidence interval 12.6-56.6%) by intention-to-treat. We conclude that GAP is an active and tolerable treatment combination, with minimal visceral organ toxicities. PMID- 16317295 TI - Irinotecan (CPT-11) combined with bolus 5-fluorouracil/leucovorin (Saltz regimen) as first-line chemotherapy of patients with advanced colorectal cancer. AB - Concerns about the safety of irinotecan (CPT-11) plus bolus 5-fluorouracil (5 FU)/leucovorin (LV) (the so-called Saltz regimen) have been previously reported. This prospective, multicenter, non-randomized study evaluated the anti-tumoral effect and toxicity of the Saltz regimen as first-line chemotherapy of 130 patients with advanced colorectal cancer (CRC). The median numbers of treatment cycles and infusions received per patient were 3 and 12, respectively. Eight (6.1) and 37 patients (28.5%) showed complete and partial responses, respectively [overall response rate=34.6% (95% confidence interval=20.7-48.5%)]. After a median follow up period of 9 months, 70 patients had died. The median progression free survival and overall survival were 6.78 (0.3-33.8) and 8.26 months (range 0.3-33.8), respectively. The combined CPT-11/5-FU/LV treatment was well tolerated and no toxic deaths were reported. The most common grade 3/4 hematological toxicity was neutropenia (28% of patients and 3% of infusions), but no febrile neutropenia was reported. Delayed diarrhea was the most reported grade 3/4 non hematological toxicity (21% of patients and 2% of infusions). Other non hematological toxicities showed very low incidences. During the study five patients died due to factors not associated with disease progression. We conclude that the Saltz regimen administered on an outpatient basis was safe and well tolerated in patients with advanced CRC. Close monitoring of external patients together with an early treatment of toxicity was found to be essential to prevent severe and potentially fatal gastrointestinal or thromboembolic events previously reported with this CPT-11 combined regimen. PMID- 16317296 TI - Epidermal growth factor receptor tyrosine kinase inhibitor, erlotinib, and concurrent 5-fluorouracil, cisplatin and radiotherapy for patients with esophageal cancer: a phase I study. AB - This phase I trial investigates the safety of combining radiation, 5-fluorouracil (5-FU) and cisplatin with the epidermal growth factor receptor tyrosine kinase inhibitor, erlotinib, in patients with esophageal carcinoma. From April 2000 to January 2005, 11 patients with squamous or adenocarcinoma of the esophagus were enrolled. Patients received either 50, 100 or 150 mg oral erlotinib/day beginning on the first day of radiation (three patients in each dose cohort). Concurrent cisplatin (75 mg/m2 i.v., days 8 and 36) and 5-FU (1000 mg/m2 i.v., days 8-11 and 36-39) were also given with 50.4 Gy thoracic radiation, delivered at 180 cGy/day, 5 days/week. Toxicity was evaluated using the National Cancer Institute Common Toxicity Criteria (version 3.0). Erlotinib with concurrent 5-FU, cisplatin and thoracic radiation was well-tolerated at 50, 100 and 150 mg/day. The major toxicities were diarrhea (grade 1=18%, grade 2=18%), skin rash (grade 4=54.5%), nausea (grade 1=18%, grade 2=54%, grade 3=9%) and dehydration (grade 3=27%). All patients experienced esophagitis during treatment (grade 1=55%, grade 2=32%, grade 3=9%, grade 4=9%). Two patients were discontinued from the study secondary to non-erlotinib-related toxicities. We conclude that the phase I study demonstrates the safety and tolerability of erlotinib delivered at 150 mg/day with concurrent 5-FU, cisplatin and thoracic radiation. The major toxicities encountered were grade 1-2 diarrhea, grade 1 skin rash, grade 1-3 nausea and grade 3 dehydration. A phase II study is planned. PMID- 16317297 TI - Delayed oxaliplatin-associated neurotoxicity following adjuvant chemotherapy for stage III colon cancer. AB - Oxaliplatin-containing chemotherapy regimens are utilized commonly for metastatic colorectal cancer and increasingly in the adjuvant setting following surgical resection. The dose-limiting toxicity is neurotoxicity. Acute neurotoxicity is cold induced and transient. Chronic neurotoxicity usually has a predictable clinical course. It is manifested by paresthesias and dysesthesias of gradually prolonged duration that occur between treatment cycles, and increase in intensity and duration with the cumulative dose. We report here a case of a patient who developed significant grade 3 chronic neuropathy following completion of 6 months of adjuvant oxaliplatin-containing chemotherapy for stage III colon cancer. The neurotoxicity was not preceded by any transient symptoms characteristic of chronic oxaliplatin neuropathy and its onset was unpredictable. Delayed neurotoxicity is a complication which must be considered for patients receiving adjuvant therapy and attempts to utilize the minimum effective cumulative dose of oxaliplatin are warranted. PMID- 16317298 TI - Gemcitabine-related radiation recall in a patient with pancreatic cancer. AB - Radiation recall refers to inflammatory reactions triggered by chemotherapeutic agents and develops cutaneously in the previously irradiated areas. Such agents include anthracyclines, taxanes and capecitabine. Radiation recall related to gemcitabine has been reported in lung and breast cancer. Similar phenomenon associated with gemcitabine, the only FDA-approved drug for pancreatic cancer, is rarely reported. We report a patient with inoperable pancreatic cancer who developed gastrointestinal bleeding secondary to radiation-recall related to gemcitabine and review literature. A 57-year-old white male with unresectable pancreatic cancer received capecitabine in combination with radiation therapy followed by capecitabine alone given over approximately a 3-month time period. Computed tomography re-evaluation demonstrated a new liver lesion. The patient was then treated with gemcitabine and irinotecan. On day 15 of cycle 1, he reported progressive worsening of weakness and fatigue, and melena. Physical examination revealed hypotension (84/47 mmHg) and heme-positive stool on rectal examination. He denied aspirin or non-steroidal anti-inflammatory drug use. Chemotherapy was held. Hematocrit was 20% (previously 33%). He was transfused with 3 units of packed red blood cells. An esophago-gastro-duodenal examination was performed which showed antritis and duodenitis consistent with radiation therapy. A single site of oozing was injected with epinephrine. The diffuse gastritis was aggressively treated with proton pump inhibitors. The patient's hematocrit eventually stabilized and was 30% at discharge. Gemcitabine was not resumed. Radiation recall from gemcitabine is rare, but can potentially arise in any site that has been previously irradiated. Gemcitabine should be added to the list of drugs known to cause radiation recall. Treating physicians must be aware of this potential toxicity from gemcitabine either given concomitantly or followed by radiation. We suggest discontinuing gemcitabine if radiation recall is observed. Further studies are warranted into the pathogenesis of this unique phenomenon. PMID- 16317299 TI - Chromosome 7 aberrations in a young girl with myelodysplasia and hepatoblastoma: an unusual association. AB - We report a 30-month-old female with intrauterine growth retardation, postnatal failure to thrive, pancytopoenia and myelodysplasia with monosomy 7 in the marrow. The child succumbed to overwhelming sepsis, following a bone marrow transplant to facilitate chemotherapy for metastatic hepatoblastoma--a tumour that has not been previously reported in myelodysplasia syndromes. Cytogenetic, molecular and microarray analysis of peripheral blood, skin fibroblasts and bone marrow revealed unusual results, suggestive of somatic chromosome instability. A normal peripheral blood karyotype was documented in infancy. Monosomy 7 was found in the bone marrow. Molecular (microsatellite marker) results for a later peripheral blood specimen were suggestive of partial maternal isodisomy 7q, and this was supported by microarray data on single-nucleotide polymorphisms. Microarray data on gene copy number, collected for the same blood specimen, indicated cryptic mosaicism for the monosomy 7 cell line, with the monosomic line lacking the paternal copy. In fibroblasts, cytogenetic data showed mosaic partial trisomy for distal 7p. PMID- 16317300 TI - Supernumerary marker chromosome 7 and maternal uniparental disomy 7 in a boy with growth retardation and triangular face. AB - We report on a 12-month-old boy with pre and post-natal growth retardation, triangular face and mild psychomotor retardation. Karyotyping revealed a supernumerary marker chromosome (SMC) in 36% of cells. Using fluorescence in situ hybridization and BAC clones, the supernumerary marker chromosome was found to be a highly deleted chromosome 7 with breakpoints within the pericentric euchromatin (partial trisomy 7). Microsatellite typing indicated maternal uniparental disomy of chromosome 7 (matUPD7). The patient showed only few signs of the Silver Russell syndrome. He therefore reflects the mild end of the phenotypic spectrum in patients with matUPD7. The case supports the argument that karyotyping is warranted in patients with short stature and only few additional features, and that UPD studies are required in patients with a SMC(7). Considering that the SMC(7) contributed very little, if any, to the phenotype of this boy, we propose that UPD7 studies should be carried out in children with pre- and postnatal growth retardation of unknown cause even in the absence of a SMC. PMID- 16317301 TI - Mosaic 13q13.2-ter deletion restricted to tissues of ectodermal and mesodermal origins. AB - The '13q-' syndrome shows widely variable manifestations. Investigation of the involvement of different tissues has never been reported in patients with 13q- syndrome previously. We describe a patient with mosaicism for del(13q) and clinical features of 13q- syndrome. The mother of the patient was professionally exposed to aniline colorants and glue components during the whole pregnancy. The patient had dysmorphic features, skeletal anomalies and brain malformations with agenesis of the corpus callosum, vermian hypoplasia and IVth ventricular system abnormalities. Eye examination revealed chorioretinal coloboma and irregular dispersion of retinal pigment in the right eye. The karyotype analyses and the molecular studies performed on peripheral lymphocytes, oral swab and cells of urinary tract were normal whereas a deletion of the long arm of chromosome 13 (13q13.2) was found in skin fibroblasts and in hair cells. We hypothesized that the 13q deletion arose during the third week after conception possibly due to a teratogenic effect and that tissue of mesodermal and ectodermal origin are involved. We suggest analysing a fibroblast karyotype when a diagnosis of 13q- syndrome is suspected on clinical ground. The role of teratogens in causing this type of mosaic chromosome abnormality also warrants further investigation. PMID- 16317302 TI - Duplication of distal 20q: clinical, cytogenetic and array CGH. Characterization of a new case. AB - Trisomy of the long arm of chromosome 20 is rare. We describe an 18-month-old male who was born at 36 weeks via Caesarian section after an uneventful pregnancy. During the newborn period he was found to have a right-sided cleft lip and cleft palate, hypertelorism, strabismus and mildly over-folded ears with cupping. Cardiovascular examination was consistent with the diagnosis of severe aortic coarctation, which was confirmed by echocardiogram. Additionally, hypothyroidism was diagnosed. Neurological evaluation at 18 months revealed a hypotonic infant with delayed acquisition of motor milestones. Cytogenetic analysis showed additional material on the long arm of chromosome 20, confirmed by fluorescence in situ hybridization (FISH) analysis as being of chromosome 20 origin. Because of the indistinct GTG-banding pattern it was not possible to distinguish between a proximal [dup(20)(q11.2q13.1)] or distal duplication [dup(20)(q13.1q13.3)]. To further define the duplication we used array comparative genomic hybridization (CGH) which demonstrated a 7.8 Mb interstitial duplication in distal 20q. Thus, the proband's karyotype was interpreted as 46,XY,dup(20)(q13.2q13.2). The proband is the first reported case of a pure duplication of this region. This case further highlights the utility of array CGH in characterizing aneusomies and, in particular, for accurate breakpoint designation and quantitation of ambiguous rearrangements. PMID- 16317303 TI - Trisomy 10p with clinical features of facio-auriculo-vertebral spectrum: a case report. AB - We report a male child born with complete absence of his external ear, hemifacial microsomia of the right side, high arched palate, a down-turned upper lip and slightly up-slanting palpebral fissures. The features were suggestive of facio auriculo-vertebral spectrum. Investigations showed a tandem duplication of the short arm of one chromosome 10 with apparent breakpoints at p14 and p15. This case extends the list of chromosomal abnormalities associated with the facio auriculo-vertebral phenotype and also adds useful clinical information to possible trisomy 10p phenotypes. PMID- 16317304 TI - Transient progeroid phenotype and lipodystrophy in mosaic polyploidy. AB - Wiedemann-Rautenstrauch syndrome is a rare disorder with a progressive course and early lethality. Severe mental and growth retardation, muscle hypotonia, a progeroid face, wrinkled skin, relative macrocephaly with late closure of the anterior fontanel, arachnodactyly and congenital heart defects are also typical. We report on a female infant with all the characteristic features of this syndrome after birth. Chromosomal studies on peripheral leukocytes showed a normal karyotype. In view of an abnormal lipid distribution and lipodystrophy, metabolic studies for congenital disorders of glycosylation have been performed with normal results. At the age of 2 years 6 months the progeroid signs were no longer present, and the patient had a striking improvement in her psychomotor development. As there are overlapping features in Wiedemann-Rautenstrauch syndrome and in mosaic polyploidy, including psychomotor retardation, reduced peripheral muscle bulk, arachnodactyly and lipodystrophy, chromosome analysis was performed in the fibroblast culture of our patient. A mosaic triploidy/tetraploidy was detected in 60% and 14% of the cells, respectively. We therefore recommend chromosome analysis of fibroblasts from patients with a neonatal presentation of progeroid features and lipodystrophy. PMID- 16317305 TI - Description of a new syndrome: auricular abnormalities and cleft lip in two sibs, in association with cleft palate and unusual opthalmological findings. AB - We describe the association of auricular abnormalities and cleft lip with or without cleft palate in two siblings. One sibling has postauricular pits, profound myopia, nystagmus and retinal pigment abnormalities. The second sibling was a fetus of 23 weeks gestation with severe cleft lip, cleft palate and external ear abnormalities. As this constellation of features has not been described together before, we believe this is a new syndrome. PMID- 16317306 TI - A case of congenital jejunal atresia associated with bilateral athelia and choanal atresia: new syndrome spectrum. AB - We present a case of bilateral choanal atresia, cleft of the posterior palate. Cardiac, respiratory and central nervous system examinations were normal. An examination of the chest revealed aplasia of the bilateral absence of the bilateral nipple-areola complex. Skin, soft tissue and bilateral pectoral muscles of the chest wall were normal. The patient did not have scalp nodules or abnormal ears. Four hours after birth, an X-ray examination showed the distended intestinal loops, suggesting small bowel obstruction. Exploration was performed and congenital jejunal atresia was surgically identified. A primary anastomosis was performed. At 3 years of age, the patient had mild mental and growth retardation. Chromosome analysis was normal (46,XX). Magnetic resonance imaging showed no abnormal finding in the spinal cord; however, mild atrophy of the left cerebrum was pointed out. An abdominal echo examination showed no abnormal findings in the kidney. These associations may represent a previously undescribed syndrome spectrum. PMID- 16317307 TI - Cerebro-oculo-facio-skeletal syndrome complicated by congenital ichthyosis. AB - We report a male patient with typical features of cerebro-oculo-facio-skeletal syndrome, diagnosed on the basis of the characteristic facies, joint contractures and microcephaly, but complicated by congenital ichthyosis on his face, trunk and limbs. PMID- 16317308 TI - Pseudotrisomy 13 syndrome: a case with left ventricular hypoplasia and duodenal stenosis. AB - We report a case of a female child born at 32 weeks of gestation. Birth weight was 1200 g (<3rd centile), length 40 cm (10th-50th centile) and head circumference 23.5 cm (<3rd centile). Clinical examination revealed microcephaly, hypotelorism, microphthalmia, a flat rudimentary nose with a single nasal cavity, high palate, thick dysplastic low-set ears, a short neck, postaxial polydactyly of the upper limbs, and single palmar creases. Investigations showed alobar holoprosencephaly, absence of the third ventricle and midline structures of the brain, microphthalmia, hypotelorism, left ventricular hypoplasia, a large atrial septal defect, and duodenal stenosis. The karyotype was 46,XX. A hypoplastic left ventricle and duodenal stenosis have not been previously reported in pseudotrisomy 13 and this case might aid in the further delineation of this syndrome. PMID- 16317309 TI - Another patient with pulmonary hypoplasia, microphthalmia and diaphragmatic hernia. PMID- 16317310 TI - Facial asymmetry with colon atresia. AB - This case report describes an unusual combination of birth defects in a black South African child presenting with colon atresia, facial symmetry due to facial microsomia and brain abnormalities. The suspected aetiology is thought to be vascular disruption in the late embryonic period. PMID- 16317311 TI - A placebo-controlled, randomized withdrawal study of sertraline for major depressive disorder in Japan. AB - The objective of this double-blind, placebo-controlled, multicentre randomized withdrawal study was to evaluate the efficacy and tolerability of sertraline for 16 weeks in treating Japanese patients with major depressive disorder (MDD) who had achieved a response to 8 weeks of sertraline treatment. Patients (n=361) were initially treated with 8 weeks of open-label sertraline treatment followed by 16 weeks of double-blind treatment with either sertraline (50-100 mg/day) or placebo. Responders during the open-label phase were eligible to be entered into the double-blind phase. A total of 235 patients (65.1%) were entered to the double-blind phase and randomly assigned to receive sertraline (n=117) or placebo (n=118). A significantly (P=0.016) lower relapse rate was found for sertraline (8.5%) compared to placebo (19.5%) during the double-blind phase. Examination of time-to-relapse showed that the relapse free rate curve was significantly higher for sertraline (log-rank test, P=0.026) than placebo. Mean changes from beginning to end of the double-blind phase on measure of depressive symptoms, quality of life and global improvement also significantly favoured sertraline over placebo. Sertraline was well-tolerated, with similar adverse events as found in previous studies. These results confirm the efficacy of sertraline in preventing the relapse of MDD in Japanese patients. PMID- 16317312 TI - An open-label extension trial of risperidone monotherapy in the treatment of bipolar I disorder. AB - The primary objective of this 9-week open-label extension trial was to assess the effects of risperidone monotherapy in patients with acute bipolar I disorder who completed treatment in two preceding 3-week double-blind trials. Patients with DSM-IV bipolar I disorder, experiencing an acute manic episode, received a flexible dose of risperidone (1-6 mg/day) or placebo in two independent double blind, randomized, 3-week monotherapy trials. Completers who required ongoing treatment were eligible to enter this open-label 9-week extension trial during which all patients received risperidone. The primary efficacy measure was the mean change in the Young Mania Rating Scale (YMRS) total score. Secondary efficacy measures included the Clinical Global Impressions-Severity Scale, Montgomery-Asberg Depression Rating Scale, Positive and Negative Syndrome Scale and Global Assessment Scale. Safety assessments included adverse event reports, laboratory tests, and the Extrapyramidal Symptom Rating Scale (ESRS). Of the 283 patients who entered the extension study, 160 had previously received risperidone (RIS/RIS) in the acute treatment trial and 123 had received placebo (PLA/RIS). This study was completed by 71% of these patients. The mean+/-SE modal dose of risperidone was 4.6+/-1.5 mg/day. Patients in both the RIS/RIS and PLA/RIS groups improved significantly at the endpoint of the 9-week open-label study compared to their open-label baseline scores (-5.2+/-0.69, P<0.001 and -9.12+/-1.44, P<0.001, respectively) on the YMRS. Furthermore, changes from double-blind baseline to open-label endpoint were -29.4+/-1.0 in the RIS/RIS group and -23.9+/-1.4 in the PLA/RIS group. Significant improvements from both double-blind and open-label baseline were seen at week 1 of the open-label trial (P<0.001) and at each subsequent timepoint. A similar pattern was observed on the secondary measures of efficacy. Most frequent adverse events were extrapyramidal disorder (18%) and somnolence (12%). Most adverse events were mild or moderate in severity. The mean score for the Parkinsonism subscale of the ESRS was 1.1 at open-label baseline, and decreased by 0.1 at endpoint. Mean increase in body weight from open-label baseline was 0.6 kg in patients treated with placebo in the preceding double blind trial and 1.2 kg in patients previously treated with risperidone. Risperidone treatment was well tolerated and resulted in further improvement during the 9-week extension, beyond the 3 weeks of acute treatment. Patients switched from placebo to risperidone improved markedly. Risperidone treatment did not induce depression. PMID- 16317313 TI - Efficacy and tolerability of ziprasidone in patients with treatment-resistant schizophrenia. AB - This multicentre, parallel-group study compared the efficacy and tolerability of ziprasidone and chlorpromazine in treatment-resistant schizophrenia (at least three treatment periods of at least 6 weeks each with two or more antipsychotic agents during the past 5 years without significant response) that was unresponsive to 6 weeks of open-label haloperidol (30 min (z = -2.3, p = .02). Not receiving eptifibatide (Integrilin) contributes to less ecchymosis (z = -2.9, p = .004). CONCLUSIONS: Nurses need to be vigilant when caring for patients who have a VC prior to femoral sheath removal and have received antiplatelet medications. Further research is needed to identify patient characteristics that may contribute to VCs. PMID- 16317361 TI - Application of the CuSum technique to evaluate changes in recruitment strategies. AB - BACKGROUND: While lagging subject enrollment in longitudinal clinical trials is a complex problem, the best recruitment strategy has not been established. Cumulative summation (CuSum) is a statistical process control procedure often applied in quality improvement efforts to detect trend shifts in highly variable serial data. OBJECTIVES: To describe changes in efforts to increase referrals and enrollment in a longitudinal quality-of-life breast cancer study, determine effects of changes in referral strategies on enrollment using a novel application of CuSum, and discuss implications of CuSum as a tool for prospectively managing the subject recruitment process. METHOD: Ten referrals and eight enrollments per month for a total of 31 months were estimated to meet study subject accrual requirements in the clinical trial. The estimates were used as standards in performing CuSum calculations. CuSum was applied to monthly referral and enrollment data and trend graphs were generated. Alterations in recruitment tactics and strategies were evaluated as to whether changes in trend occasioned such alterations. Unplanned changes in trend were noted. RESULTS: While monthly data were highly variable, an average of 8.42 referrals and 5.92 enrollments were realized during Months 1-12. Based on these figures, projected accrual for 31 months would have enrolled only 184 subjects, 66 subjects short of target. CuSum illustrated this trend. Subsequent shifts in enrollment trends were shown with improvements in referral. DISCUSSION: Indications for use of CuSum include (a) earlier detection of enrollment trend shifts, and (b) earlier discrimination between effective and ineffective recruitment. Thus, CuSum has implications for both evaluating the effects of planned and unplanned process changes and for managing the recruitment process. PMID- 16317362 TI - Multilevel modeling of a clustered continuous outcome: nurses' work hours and burnout. AB - BACKGROUND: Multilevel models were designed to analyze data generated from a nested structure (e.g., nurses within hospitals) because conventional linear regression models underestimate standard errors and, in turn, overestimate test statistics. OBJECTIVES: To introduce 2 types of multilevel models, the random intercept model and the random coefficient model, to describe the correlation among observations within a cluster, and to demonstrate how to identify the superior model. METHOD: The conceptual and mathematical bases for the 2 multilevel model types are presented. Intraclass correlation is defined and assessment of model fit is detailed. An empirical example is presented in which average work hours per week and burnout are analyzed using data from 4,320 staff nurses clustered in 19 hospitals. RESULTS: Average work hours were positively associated with nurse burnout. The multilevel models corrected the problem of underestimated standard errors in conventional linear regression models. Graphs displaying the hospital-level differences illustrated the 2 multilevel model types. Although the multilevel models corrected the underestimation of standard errors, the results did not differ substantively for the conventional or the 2 multilevel models. The intraclass correlation coefficient was .044, indicating that the extent of shared variance among nurses in a hospital was low. The random intercept model fit the data better than did the random coefficient model. CONCLUSIONS: Multilevel models provide a more accurate and comprehensive description of relationships in clustered data than do conventional models, by correcting underestimated standard errors, by estimating components of variance at several levels, and by estimating cluster-specific intercepts and slopes. PMID- 16317363 TI - Evaluating content validity for children's self-report instruments using children as content experts. AB - BACKGROUND: The development and evaluation of instruments to index cognitive and emotional processes from the perspectives of children is a priority for pediatric nursing research. OBJECTIVE: To describe the procedures used in employing children as content validity experts in the development of a self-report instrument. METHODS: Following published recommendations for moving from qualitative research to quantitative measurement with adults and for maximizing content validity in self-report instruments, six children aged 8-16 years undergoing treatment for cancer constituted the panel of content experts for review of a measure of children's illness-related uncertainty derived from qualitative interviews. Children were provided with an explanation of the project, an explanation of their role as experts, and explicit instructions on how to evaluate the representativeness of individual items and the total scale. RESULTS: Generally, the children performed the review tasks effectively, although two children (ages 8 and 16 years) had initial difficulty in going beyond their own experience when considering the relevance of individual items. Twenty items were deemed acceptable by at least five out of the six children and two additional items were revised based on their input. DISCUSSION: Employing children as content validity experts adds a critical dimension to establishing psychometrically sound measures for studying the processes affecting the health of children and families. PMID- 16317368 TI - Perspective: why was our academy formed? PMID- 16317369 TI - Reliability and consistency of visual acuity and contrast sensitivity measures in advanced eye disease. AB - PURPOSE: The purpose of this article is to establish reliability of standard visual function measures in patients with severe vision loss. METHODS: Legally blind subjects were enrolled in a repeated-measures study to determine the reproducibility of a battery of psychophysical vision measures. Tests reported here include ETDRS visual acuity (VA) under regular and dim illumination and Pelli-Robson letter contrast sensitivity (CS) under regular, dim, and glare conditions. Two to five test repetitions were performed at monthly intervals. Subject groups included retinitis pigmentosa (RP) (n = 29), macular disease (MD) (n = 17), optic nerve disease (ON) (n = 4), diabetic retinopathy (DR) (n = 5), and other retinal diseases (OR) (n = 10). RESULTS: For most subject groups, VA measures had mean 95% coefficient of repeatability (CR.95) of 0.20 (group means between 0.13 and 0.36) log units, and CS measures had mean CR.95 of 0.35 (group means between 0.19 and 0.48) log units. CONCLUSIONS: The established reliability of VA and CS tests in subjects with severe visual impairments will allow investigators to determine whether a significant change in vision has occurred during future clinical trials to reverse, halt, or slow vision loss. The criterion for change needs to be larger than for normally sighted controls, i.e., approximately two to three lines for VA and between eight to 11 letters for CS. PMID- 16317370 TI - The mydriatic effect of multiple doses of tropicamide. AB - PURPOSE: We investigated whether two drops of 0.5% tropicamide produced a wider pupillary dilation when compared with a single drop and saline control in subjects with dark-colored irides. METHODS: Twelve young adult subjects with dark irides and who were free from ocular disease participated in the study. One eye of each subject, selected at random, was designated as experimental and the fellow eye as control. A single drop of 0.5% tropicamide was instilled in the lower cul-de-sac of each eye in turn. Immediately after, a second drop of 0.5% tropicamide was instilled in the experimental eye and a single drop of unpreserved saline into the control eye. A double-masked experimental protocol was followed. Digital images of both pupils of each subject were taken at baseline (predrug instillation) and then every 5 minutes up to 30 minutes postdrug instillation and at 40, 50, and 60 minutes. The images were subsequently viewed on a PC monitor, and pupil size was measured using a calibrated millimeter scale. RESULTS: Pupil diameter increased as a function of time for both conditions reaching a maximum between 20 and 30 minutes after drop instillation. Pupil diameter was greater under the experimental condition compared with the control (analysis of variance p < 0.001). Pupil diameter reached at least 6 mm under both the experimental and control conditions. CONCLUSION: In young healthy subjects, compared with a single drop of tropicamide, two drops were associated with a greater degree of pupillary dilation on average over the 60-minute study period. However, the magnitude of the difference was small and not clinically significant. A single drop of tropicamide produced a pupillary diameter of at least 6 mm, which should be sufficient for the conduct of a thorough dilated fundus examination. PMID- 16317371 TI - The frequency of dilated diabetic eye examinations by optometrists in the state of ohio. AB - PURPOSE: The purpose of this study was to assess optometric care of diabetic patients. The American Optometric Association recommends dilated eye examinations at least once a year for people with diabetes. METHODS: Onsite chart reviews of 1497 diabetic patient charts were performed at a volunteer sample of Ohio-based optometric practices (n = 82). Four percent of all optometrists in the state of Ohio were surveyed. Data were collected from a 10-year period and included doctor age, membership in the Ohio Optometric Association (OOA) Diabetes Initiative, patient age, date of examinations, chief complaints, and dilation at examination. The primary outcome of the study was the frequency of dilated eye examination for diabetic patients by optometrists. RESULTS: Optometrists dilated 85.8% of diabetic patients at their last comprehensive examination. The percentage of diabetic patients dilated by optometrists increased from 40.0% in 1994 to 91.5% in 2003. The average time since last comprehensive eye examination for diabetic patients was 1.0 +/- 1.8 (mean +/- standard deviation) years. Diabetic patients who were dilated at a comprehensive eye examination returned to care on average 0.7 years sooner than those who were not dilated. Doctor-level factors related to a higher average percentage of dilations were younger doctor age (beta = -6.1%/10 years of doctor age; 95% confidence interval [CI], -10.1 to -2.1) and membership in the OOA Diabetes Initiative (beta = 11.6%; 95% CI, 2.3 to 20.8). CONCLUSIONS: There is variability in diabetic eye care by optometrists. However, the majority of optometrists surveyed in this study provided dilated eye examinations for their diabetic patients. Programs desiring to increase compliance to diabetic standard of care guidelines may benefit by targeting both doctors and patients. PMID- 16317372 TI - The effect of multiple antireflective coatings and center thickness on resistance of polycarbonate spectacle lenses to penetration by pointed missiles. AB - PURPOSE: Previous work has shown that the impact resistance to blunt missiles is affected by coatings applied to either CR-39 or polycarbonate lenses. We investigated the effects of multiple antireflection (minimum angle of resolution [MAR]) coatings on the resistance of polycarbonate lenses to puncture on impact by sharp, high-speed missiles. METHODS: Four groups of surfaced plano polycarbonate lenses were investigated. Two groups had a scratch-resistant (SR) coating applied to both surfaces. One of these groups had a 2-mm center thickness and the other had a 3-mm center thickness. The other two groups of 2-mm and 3-mm thick lenses had a MAR coating applied over the SR coating. The lenses were impacted by a missile consisting of an industrial sewing machine needle mounted in a cylindrical aluminum carrier. RESULTS: The sharp missiles were able to pierce the lenses at speeds between 29.6 m/s and 46.2 m/s. Impact resistance was lowest for the thinner lenses and lenses with a MAR coating. The effect of the MAR and lens thickness was subadditive. CONCLUSIONS: We have confirmed previous observations that polycarbonate lenses are more susceptible to penetration by sharp, high-speed missiles than blunt missiles. We have also found that reducing lens center thickness and applying a MAR coating further reduces the penetration resistance. Therefore, the use of 2-mm center thickness and MAR-coated polycarbonate lenses should be discouraged for industrial eye protectors where sharp missile hazards are possible. PMID- 16317373 TI - An evaluation of the Mars Letter Contrast Sensitivity Test. AB - PURPOSE: The Mars Letter Contrast Sensitivity Test (initially known as the Lighthouse Letter Contrast Sensitivity Test) is similar in design to the Pelli Robson Test but may offer several advantages. This study evaluates the repeatability of the Mars test and its agreement with the Pelli-Robson test in normal and low-vision subjects. METHODS: Fifty-four subjects were tested (age 22 86 years), including 20 normally sighted young adults, 17 normally sighted older adults, and 17 adults with low vision (20/16 to 20/250). Subjects were tested with both contrast sensitivity tests and with the ETDRS visual acuity chart. After a short break, subjects were retested with an alternate form of each contrast sensitivity test. The chart forms used (two Pelli-Robson and three Mars) and the order of testing were varied systematically. Testing was monocular with habitual correction and, for subjects over 40 years of age, included appropriate near add. Letter-by-letter scoring was used for both tests. Repeatability and agreement were assessed by determining the 95% limits of agreement (LoA): +/- 1.96 standard deviations of the differences between administrations or tests. RESULTS: The Mars test showed excellent agreement with the Pelli-Robson test, with 95% LoA of +/- 0.21 log units for all subjects. The Mars test was similarly repeatable (95% LoA = +/-0.20 log units) to the Pelli-Robson test (95% LoA = +/ 0.20 log units) among all subjects. CONCLUSION: The new Mars Letter Contrast Sensitivity Test shows excellent agreement with the Pelli-Robson test and has similar repeatability. There are subtle differences in the actual contrast levels on different forms of the Mars test, and adjusting for these differences leads to superior repeatability of the Mars test. Thus, the Mars test may be a useful alternative to the Pelli-Robson test offering several advantages, including smaller size, improved durability, and ease of use. PMID- 16317374 TI - A method for differentiating ocular higher-order aberrations from light scatter applied to retinitis pigmentosa. AB - PURPOSE: The purpose of this study is to report a method for differentiating ocular higher-order aberrations and intraocular light scatter based on a deconvolution technique. METHODS: An optical system was used to image a laser slit on the retina and also to perform Shack-Hartmann wavefront sensing. From the laser slit image, the line spread function, incorporating both ocular higher order aberrations and light scatter, was derived. The laser slit image was deconvolved with a point spread function obtained from the Shack-Hartmann image. The area under the line spread function that was derived from the laser slit image after deconvolution provided a measurement of intraocular light scatter. The deconvolution technique was applied to images obtained in a group of 13 patients (mean age +/- 1 standard deviation: 42 +/- 12 years) with retinitis pigmentosa (RP), a retinal disease in which, by clinical examination, changes in the lens of the eye can be manifested. Measurements were compared with those obtained from 20 visually normal control subjects (mean age +/- 1 standard deviation: 43 +/- 17 years). RESULTS: Combined higher-order aberrations and light scatter, measured as the area under the line spread function derived from the laser slit image, were increased significantly in the patients with RP as compared with the control subjects (p = 0.004). Ocular higher-order aberrations obtained from the Shack-Hartmann images were higher in the patients with RP than in the control subjects (p = 0.05). Intraocular light scatter derived from the deconvolved laser slit image was significantly higher in the patients with RP than in the control subjects (p = 0.009). Minimizing the contribution of ocular higher-order aberrations by deconvolution reduced the area under the line spread function in the control subjects and patients with RP, denoting an improvement in retinal image quality. CONCLUSIONS: A method for differentiating ocular higher order aberrations and intraocular light scatter based on deconvolution was developed that may be useful for determining the level of improvement in retinal image quality that can be anticipated by the application of adaptive optics to aging and diseased human eyes. PMID- 16317375 TI - Advantages of terminating Zippy Estimation by Sequential Testing (ZEST) with dynamic criteria for white-on-white perimetry. AB - PURPOSE: A number of automated perimeters use the Zippy Estimation by Sequential Testing (ZEST) algorithm, which is an adaptive Bayesian method, for determining sensitivity measures. There are two popular rules for deciding when to terminate Bayesian procedures: (1) after a fixed number of presentations; or (2) when the probability density function (pdf) over all thresholds modified by the procedure becomes sufficiently narrow (a dynamic termination criterion). It has recently been argued that fixed termination criteria perform equally as well as dynamic criteria when applied in a fashion typical of laboratory-based visual psychophysics. Perimetry, however, has specific requirements; the tests must be very short, there is a wide range of possible sensitivities, and erroneous responses from the patient must be tolerated. This study used computer simulation to compare fixed and dynamic termination criteria for the ZEST algorithm using conditions typical of white-on-white perimetry. METHODS: Eight ZEST procedures were compared using the following termination criteria: fixed termination after 4, 5, 6, 7, and 8 presentations; dynamic termination when the standard deviation of the pdf was 1 dB, 1.5 dB, and 2 dB. Four patient error models were used: ideal, typical false-positive, typical false-negative, and unreliable patients. We also ran a version of ZEST that set the likelihood function exactly equal to the patient's frequency of seeing curve. RESULTS: The mean absolute error and standard deviation of error in threshold measurement was higher for the fixed termination criteria than for dynamic termination criteria of the same average number of presentations. CONCLUSIONS: The results of our simulations indicate that dynamic procedures have some distinct benefits over fixed termination procedures when a minimum of presentations are required and response errors are made as in a white-on-white perimetric setting. Dynamic termination criteria are at least partially successful in expending more presentations when required to enhance test precision. PMID- 16317376 TI - Horizontal fixation disparity measures with nonius lines. AB - PURPOSE: It has been reported that changes in visual direction and retinal correspondence may limit the validity of nonius lines as a subjective measure of vergence, at least in particular viewing conditions, e.g., dynamic or forced vergence. Nonius lines may be valid at larger spatial separation between fusion contour and nonius lines. Therefore, we measured fixation disparity varying the amount of a vertical gap between nonius lines. METHODS: A static central fusion stimulus was given at a 100-cm viewing distance. The nonius lines were presented with various vertical gaps either continuously (and adjusted to subjective alignment) or flashed in a series of trials (in adaptive testing). Twelve subjects with normal binocular vision were tested repeatedly to identify individual vergence characteristics. RESULTS: When the vertical gap between nonius lines was increased up to 7.9 deg, the amount of subjective fixation disparity tended to increase in part of the subjects, predominantly in those with an exo fixation disparity. Most subjective fixation disparity measures were correlated with each other and with tonic vergence (i.e., dark vergence tested subjectively without fusion stimulus); however, flashed nonius lines at larger nonius gaps were an exception. CONCLUSIONS: We found physiologically plausible measures of idiosyncratic fixation disparity with continuous nonius lines at any amount of nonius gap or with flashed nonius lines at small gaps. In these conditions, the intersubject variability of fixation disparity was much larger than effects of the spatial separation between fusion stimulus and nonius lines. PMID- 16317377 TI - What's new in shock, December 2005. PMID- 16317378 TI - From the experimental laboratory to the patient: how does it happen? AB - Many potential therapeutic agents or drugs are evaluated in experimental animal laboratories, but in spite of interesting effects, they infrequently come into common clinical use. The reasons for this are reviewed. Agents studied previously and many being studied now are cited. Perhaps the biggest benefit of study of drugs in animals (other than for safety and toxicity) is to help us better understand the pathophysiology of disease. PMID- 16317379 TI - Vascular endothelial growth factor is increased during the first 48 hours of human septic shock and correlates with vascular permeability. AB - Meningococcal septic shock is an important cause of morbidity and mortality in children and young adults worldwide and is the prototypical gram-negative septic shock. One of the key factors in the development of shock is increased microvascular permeability. Vascular endothelial growth factor (VEGF) is a central factor in angiogenesis and is an important mediator of vascular permeability. Thirteen patients with meningococcal infection (eight presenting with shock) were investigated in the early phase of invasive meningococcal disease. Cytokines, complement activation, and VEGF plasma concentrations were measured during the first 48 h on the pediatric intensive care unit. Increased cytokine concentrations and activation of the complement system were observed. VEGF plasma concentrations were increased (median 193 pg/mL, range 71-1082) and were highest in the presence of shock (208 pg/mL, 169-1082) compared with patients presenting without shock (92 pg/mL range 71-299). VEGF concentration at admission correlated with the severity of disease (pediatric risk of mortality score, R=0.90 [Spearman], P=0.0001) and the amount of fluids administered within the first 24 h (R=0.90, P<0.0001). In all patients, a decrease in VEGF was associated with a decrease in fluid intake during t=24 to 48 h. The results suggest that apart from correlation with IL-1 beta, -10, -12, and complement activation, microvascular permeability in sepsis is also closely linked to the plasma concentration of VEGF. The role of VEGF in sepsis-associated increased microvascular permeability needs further exploration and may represent a new therapeutic target. PMID- 16317380 TI - Diagnostic accuracy of left ventricular function for identifying sepsis among emergency department patients with nontraumatic symptomatic undifferentiated hypotension. AB - The hypothesis of this study states that in emergency department (ED) patients with non-traumatic symptomatic hypotension, the presence of hyperdynamic left ventricular function (LVF) is specific for sepsis as the etiology of shock. We performed a secondary analysis of patients with non-traumatic symptomatic hypotension enrolled in a randomized, clinical diagnostic trial. The study was done in an urban tertiary ED with a census over 100,000 visits per year. Inclusion criteria were non-trauma ED patients aged >17 years, initial vital signs consistent with shock (systolic blood pressure <100 mm Hg or shock index >1.0), and agreement of two independent observers for one sign and symptom of circulatory shock. All patients underwent focused ED echocardiography (echo) during initial resuscitation. Echos were reviewed post-hoc by a blinded physician and categorized by qualitative LVF as hyperdynamic (ejection fraction [EF] >55%), normal to moderate impairment (EF 30%-55%), and severe impairment (EF <30%). Main outcome was the criterion standard diagnosis of septic shock. Analyses include the diagnostic performance of LVF, Cohen's kappa for interobserver agreement of LVF, and logistic regression for independent predictors of sepsis. There were 103 echos that were adequate for analysis. The mean age was 57+/-16.7 years, 59% were male, and the mean initial systolic blood pressure was 83+/-11.3 mm Hg. A final diagnosis of septic shock was made in 38% (39/103) of patients. Seventeen of 103 (17%) patients had hyperdynamic LVF with an interobserver agreement of kappa=0.8. The sensitivity and specificity of hyperdynamic LVF for predicting sepsis were 33% (95% CI 19%-50%) and 94% (85%-98%), respectively. Hyperdynamic LVF had a positive likelihood ratio of 5.3 for the diagnosis of sepsis and was a strong independent predictor of sepsis as the final diagnosis with an odds ratio of 5.5 (95% CI 1.1-45). Among ED patients with non-traumatic undifferentiated symptomatic hypotension, the presence of hyperdynamic LVF on focused echo is highly specific for sepsis as the etiology of shock. PMID- 16317381 TI - Genetic predisposition for a compromised immune system after multiple trauma. AB - Severe trauma induces sustained changes of the immune response, which are thought to be related to secondary organ dysfunction. Despite a similar injury severity, the extent of the inflammatory response may vary between polytraumatized patients. It is unclear whether inflammatory variability is associated with genetic variations. In this prospective cohort study, patients were included when the following criteria were fulfilled: Injury Severity Score >16, age 18 to 60 years, and a survival >48 h after injury. Four different polymorphisms (TNF-Nco1, IL-1-Taq1, IL-6-174G/C, and IL-8-251A/T) were determined. Patients were separated according to the severity of the systemic inflammatory response syndrome (SIRS; ACCP/SCCM criteria: >2 criteria at 2 consecutive days or at 3 days of the observation period: group +SIRS; /=70 years) with small cell lung cancer. AB - Platinum-based treatment for small cell lung cancer (SCLC) has been established since 1995. This study investigates treatment outcome of elderly patients (age >/=70 years) with SCLC over the past 20 years in a large UK cancer centre. Comparison of all-cause survival was assessed in patients presenting between two predefined time periods: 1982-1994 and 1995-2003. All the survival analysis were adjusted for stage and performance status and age if appropriate. Survival between different chemotherapy treatment regimens was compared. A total of 322 elderly patients (31% of all) registered between 1982-2003 received chemotherapy for SCLC. Patients presenting in 1995-2003 had an overall better median survival (43 vs 25 weeks) and a 1-year survival (37 vs 14%) than patients presenting in 1982-1994 (P<0.001). This applied to patients with both limited and extensive stage disease and all age groups. There was a trend towards the use of more platinum-based treatments in the later cohort but the use of radiotherapy remained constant. Patients who received platinum combinations (Carboplatin or Cisplatin) had significantly improved survival over those who received single agents or other combinations (P<0.001) and there was no significant difference between carboplatin and cisplatin (P=0.7). The analysis demonstrates that there has been a significant improvement in survival for elderly patients with lung cancer treated by chemotherapy in the past 20 years despite more very elderly patients being treated with a poorer performance status. This change is probably multifactorial and may be due to the increased use of platinum-based treatment and improved supportive care. PMID- 16317432 TI - Factors associated with emotional and behavioural problems among school age children of breast cancer patients. AB - To identify factors linked with emotional and behavioural problems in school age (6- to 17-year-old) children of women with breast cancer. Reports of children's emotional and behavioural problems were obtained from patient mothers, their healthy partners, the children's teacher and adolescents using the Child Behaviour Checklist and Mental Health subscale of the Child Health Questionnaire. Parents reported on their own level of depression and, for patients only, their quality of life. Family functioning was assessed using the Family Assessment Device and Cohesion subscale of the Family Environment Scale. Using a cross sectional within groups design, assessments were obtained (N=107 families) where the patients were 3-36 months postdiagnosis. Risk of problems in children were linked with low levels of family cohesion, low affective responsiveness and parental over-involvement as reported by both child and mother. Adolescents reported family communication issues, which were associated with externalising behaviour problems. Maternal depression was related to child internalising problems, particularly in girls. Whether the mother was currently on or off chemotherapy was not associated with child problems nor was time since cancer diagnosis. These findings held across child age. Where mothers have early stage breast cancer, a substantial minority of their school-aged children have emotional and behavioural problems. Such cases are characterised by the existence of maternal depression and poor family communication, rather than by the mother's treatment status or time since diagnosis. Targeted treatments, which focus on maternal depression and family communication may benefit the children and, through improved relationships, enhance the patients' quality of life. PMID- 16317433 TI - Patterns of care and survival for patients aged under 40 years with bone sarcoma in Britain, 1980-1994. AB - The purpose of the study was to calculate population-based survival rates for osteosarcoma (OS) and Ewing's sarcoma (ES) in Great Britain during 1980-1994, determine proportions of patients treated at specialist centres or entered in national and international clinical trials, and investigate effects of these factors on survival. Data on a population-based series of 1349 patients with OS and 849 with ES were compiled from regional and national cancer registries, UK Children's Cancer Study Group, regional bone tumour registries and clinical trials. Follow-up was through population registers. Survival was analysed by actuarial analysis with log-rank tests and by Cox's proportional hazards analysis. Five-year survival rates during 1980-1984, 1985-1989 and 1990-1994 were 42% (95% CI: 37, 46), 54% (95% CI: 50, 59) and 53% (95% CI: 48, 57), respectively, for OS and 31% (95% CI: 26, 37), 46% (95% CI: 40, 51) and 51% (95% CI: 45, 57) for ES. Proportions of patients treated at a supraregional bone tumour centre or a paediatric oncology centre in the three quinquennia were 36, 56 and 67% for OS and 41, 60 and 69% for ES. In 1983-1992, 48% of OS patients were entered in a national trial; for ES, 27% were entered in 1980-1986 and 54% in 1987-1994. Survival was similar for trial and nontrial patients with OS. For ES, trial patients had consistently higher 5-year survival than nontrial patients: 1980-1986, 42 vs 30%; 1987-1992, 59 vs 42%; 1993-1994, 54 vs 43%. During 1985-1994, patients with OS or ES whose main treatment centre was a nonteaching hospital had lower survival rates. In multivariate analyses of patients diagnosed during 1985-1994 that also included age, sex, primary site, surgical treatment centre, the results relating to main treatment centre for both OS and ES retained significance but the survival advantage of trial entry for ES became nonsignificant. For both OS and ES diagnosed since 1985, patients whose main treatment centre was a nonspecialist hospital had a lower survival rate. PMID- 16317434 TI - No relationship between thymidine phosphorylase (TP, PD-ECGF) expression and hypoxia in carcinoma of the cervix. AB - The expression of hypoxia-regulated genes promotes an aggressive tumour phenotype and is associated with an adverse cancer treatment outcome. Thymidine phosphorylase (TP) levels increase under hypoxia, but the protein has not been studied in association with hypoxia in human tumours. An investigation was made, therefore, of the relationship of tumour TP with hypoxia, the expression of other hypoxia-associated markers and clinical outcome. This retrospective study was carried out in patients with locally advanced cervical carcinoma who underwent radiotherapy. Protein expression was evaluated with immunohistochemistry. Hypoxia was measured using microelectrodes and the level of pimonidazole binding. There was no relationship of TP expression with tumour pO(2) (r=-0.091, P=0.59, n=87) or pimonidazole binding (r=0.13, P=0.45, n=38). There was no relationship between TP and HIF-1alpha, but there was a weak borderline significant relationship with HIF-2alpha expression. There were weak but significant correlations of TP with the expression of VEGF, CA IX and Glut-1. In 119 patients, the presence of TP expression predicted for disease-specific (P=0.032) and metastasis-free (P=0.050) survival. The results suggest that TP is not a surrogate marker of hypoxia, but is linked to the expression of hypoxia-associated genes and has weak prognostic power. PMID- 16317435 TI - The measurement and therapeutic implications of circulating tumour cells in breast cancer. AB - Circulating tumours cells (CTCs) represent an important biologic link in the spread of breast cancer from primary to metastatic disease. CTCs are strong predictors of prognosis in patients with metastatic breast cancer. Research to date has focused on development of methods with adequate sensitivity and specificity to reproducibly identify these rare events. Future research will focus on the biologic phenotypes of these cells with goals to understand mechanisms of metastasis, to identify novel therapeutic targets, and to monitor response to therapy. PMID- 16317437 TI - Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. Conclusions. AB - This paper summarises and presents in context the main findings of an extensive series of studies of early training lasting 13 weeks in which the tissue responses of 2-year-old Thoroughbred horses were assessed using a combination of methods. Negligible clinical injury was detected and thus the study fulfilled the intention of investigating adaptive change rather than injury. Cancellous and cortical bone, some digital tendons, and articular cartilage responded to early training exercise to a greater or lesser degree. Clinical examination and ancillary diagnostic aids currently in veterinary clinical use are not sufficient to detect early abnormalities in metacarpo-phalangeal joint (MCPJ) cartilage found in both trained and untrained horses. Future work should centre on detection of such changes, on the precise registration of training workload, and on the manipulation of the responses of musculoskeletal tissues by careful investigation of the effects of introducing conditioning exercise at a young age. PMID- 16317438 TI - Breeding dairy cows for the future in New Zealand. AB - A brief history of the breeding of dairy cattle in New Zealand is provided. Dairy farming in New Zealand is unique compared with the majority of dairy systems in the developed world. New Zealand has a dependence on grass-based diets and a strict requirement for a 365-day calving interval. Four main areas are discussed: future traits to evaluate, advances in genetic evaluation technologies, impacts of crossbreeding, and future progeny testing schemes. These areas are not independent, e.g. the trend of increasing numbers of crossbred cattle in the national herd will have major impacts on the design of breeding schemes. It is foreseeable that in the future there will be improvements in the national breeding goal to better reflect on-farm profitability, and in the definition of traits and methods of data capture within the national breeding goal. Methods of selection and genetic evaluation that are currently feasible for a small population will become feasible for large populations as computing power improves. Genetic improvement of cows in New Zealand will continue to be a critical component of the increased economic efficiency achieved on dairy farms in this country. PMID- 16317439 TI - Emerging technologies for identifying superior dairy cows in New Zealand. AB - The performance of animals is determined by the interaction of their genes with environmental circumstances. Accordingly, animals exhibiting superior performance are not necessarily the animals with the best genes nor are they the best choice of parents. Statistical analyses of production records for repeated traits, e.g. lactation yields and reproductive performance, show that part of the variation in performance among animals in the same herd and year is due to genetic differences, and the remainder is due to so-called residual or environmental factors that are not passed on to offspring. These within-herd environmental factors can be partitioned into a component that affects performance throughout an animal's lifetime, and a part that is unique to each observation. The process of animal evaluation from pedigree and performance records partitions the superiority of each cow into these three components. Reliable assessment of the genetic merit of bulls has required progeny testing, and for cows has required observation of their own individual performance. Selection on the genetic or breeding value component has systematically improved animal performance over recent decades, but has been limited by the age at which assessments of genetic merit are available. Emerging molecular technologies can read DNA sequences or measure RNA expression and have allowed the identification of a number of chromosome regions, and a few specific genes in those regions, that influence economic performance. This information allows better characterisation of the relationships between animals and more accurate predictions of genetic merit in bulls without progeny information and in cows that have yet to produce their own performance record. At some stage, enough genes responsible for variation in performance will be identified to allow faster genetic progress through selection of animals at young ages and therefore more rapid turnover of the generations. Mechanisms that modify gene expression have been identified and these may ultimately allow animals to be selected at an early age for lifetime productivity, accounting for processes that modify gene expression and lead to differences in performance that are not reflected by DNA sequence information. This review describes the status of these emerging technologies and their likely role in the improvement of dairy cattle. PMID- 16317440 TI - Lactational traits of importance in dairy cows and applications for emerging biotechnologies. AB - New molecular and quantitative genetic technologies are the latest in a long list of technologies that have been introduced to dairy industries over many decades to improve the performance of cows. The catalysts for future advances will be sequencing of the bovine genome and development of high-throughput technologies to identify and exploit relevant variation in DNA sequences. The new technologies will allow the selection of animals based on specific genotypes that suit specific applications such as once-daily (OAD) milking or extended lactation. These technologies will also allow exploitation of between-cow variation in milk composition, which is currently hidden by bulking of milk on and between farms. Thus, there are opportunities to produce milk from herds of cows selected for specific milk composition that might be, for example, more suitable for cheese making or have milkfat with specific properties to enhance human health. Identification of genes and gene polymorphisms associated with improved resistance to specific diseases in cows and other farm species also represents a real opportunity to improve animal health over the coming decade. New levels of genetic regulation have been identified, e.g. microRNA and epigenetics, the impacts of which on the performance of cows and humans are only just beginning to be understood. There is a potential role for veterinarians to provide or be the interface for provision of genetic advice to farmers in much the same way that nutritional advice is currently given. PMID- 16317441 TI - Non-lactational traits of importance in dairy cows and applications for emerging biotechnologies. AB - Dairy cattle have traditionally been selected for their ability to produce milk and milk components. The traditional single-minded approach to selection of dairy cattle has now changed and secondary traits are being included in selection indices by decreasing the emphasis on production. Greater emphasis on non production traits reflects the industry's desire for functional dairy cattle. Six broad categories of non-lactational traits are discussed in this review. They are: type; growth, body size and composition; efficiency of feed utilisation; disease resistance, e.g. udder health as measured by somatic cell score; reproduction; and management. Most of these traits can be found within selection indices worldwide, although relative emphasis varies. The non-lactational traits mentioned above are quantitative, meaning that the phenotype in the whole animal represents the sum of lesser traits that cannot be easily measured. The physiological mechanisms that underlie quantitative traits are extremely complex. Genetic selection can be applied to quantitative traits but it is difficult to link successful genetic selection with the underlying physiological mechanisms. The importance that the bovine genome sequence will play in the future of the genetics of dairy cattle cannot be understated. Completing the bovine genome sequence is the first step towards modernising our approach to the genetics of dairy cattle. Finding genes in the genome is difficult and scanning billions of base pairs of DNA is an imperfect task. The function of most genes is either unknown or incompletely understood. Combining all of the information into a useable format is known as bioinformatics. At the present time, our capacity to generate information is great but our capacity to understand the information is small. The important information resides within subtle changes in gene expression and within the cumulative effect that these have. Traditional methods of genetic selection in dairy cattle will be used for the foreseeable future. Most non lactational traits are heritable and will be included in selection indices if the traits have value. The long-term prognosis for genome science is good but advances will take time. Genetic selection in the genome era will be different because DNA sequence analysis may replace traditional methods of genetic selection. PMID- 16317442 TI - Vaccination of pregnant ewes against infection with Salmonella Brandenburg. AB - AIMS: To develop a challenge model for Salmonella Brandenburg infection in pregnant ewes. To compare efficacies of a live attenuated Salmonella Typhimurium mutant, a subunit preparation from a virulent S. Brandenburg isolate, and a commercial multivalent inactivated vaccine in their ability to prevent experimental S. Brandenburg infection. To assess the efficacy of the live attenuated S. Typhimurium mutant against natural S. Brandenburg infection in lambs. METHODS: Two-year-old ewes were immunised with either a live attenuated vaccine (eye-drop; n=20), a subunit vaccine (n=20) or an inactivated bacterin vaccine (n=20), both administered subcutaneously, or served as unvaccinated controls (n=21). Four weeks later, the sensitising regime was repeated as a booster vaccination, and the ewes were challenged 6 weeks later with a virulent S. Brandenburg isolate, approximately 6 weeks prior to lambing. The presence of clinical signs, abortion or death was noted following challenge. The presence and number of Salmonella spp in faecal samples taken throughout the trial, and in organs collected post mortem, were determined using an enrichment selection procedure, and confirmed by serology and pulsed-field gel electrophoresis (PFGE). Half of the surviving lambs were vaccinated with the live attenuated vaccine and all (n=39) were exposed to natural infection from contaminated pasture. RESULTS: There was no significant protection against mortality and abortion following vaccination with the live attenuated, subunit and inactivated vaccines following experimental challenge with S. Brandenburg. There was a significant but transient decrease in the number of ewes shedding S. Brandenburg (live attenuated, p=0.05; subunit, p=0.05; inactivated, p=0.01), and in the quantity of these bacteria in the sheep from the vaccinated groups (p<0.05) compared with controls, 6 weeks after challenge. Lambs from the challenged ewes did not shed Salmonella spp after being vaccinated with the live attenuated vaccine, in contrast to some of the controls, when grazed on pasture contaminated with S. Brandenburg. CONCLUSIONS: The use of live attenuated, subunit and inactivated vaccines did not significantly protect sheep against lethal experimental challenge with S. Brandenburg. PMID- 16317443 TI - Using urea dilution to standardise components of pleural and bronchoalveolar lavage fluids in the dog. AB - AIM: To develop a technique to estimate the volume of epithelial lining fluid (ELF) obtained during bronchoalveolar lavage (BAL) and pleural lavage (PL) in the dog, using the urea dilution method. METHODS: BAL and PL fluids were obtained by saline lavage of pulmonary and pleural cavities of nine clinically healthy mixed breed dogs immediately after euthanasia. Cell counts in the BAL and PL fluids were measured using standard techniques. The concentration of ELF in each lavage fluid was calculated from the relative concentration of urea in plasma and in each type of lavage fluid. Cell counts in ELF were then calculated. RESULTS: There were substantially higher cell counts in ELF compared to BAL or PF fluid. However, nucleated cell counts in ELF could not be predicted from cell counts in BAL or PL fluid. CONCLUSIONS AND CLINICAL RELEVANCE: These results suggest that accurate assessment of cellular or non-cellular components in lavage fluids should include a calculation of the proportion of ELF recovered, using a method such as urea dilution. PMID- 16317444 TI - Serological survey of pre-weaned New Zealand fur seals (Arctocephalus forsteri) for brucellosis and leptospirosis. AB - AIM: To conduct a longitudinal serological survey for evidence of Brucella spp and Leptospira spp infection of pre-weaned New Zealand fur seals in a colony on the Otago Peninsula. METHODS: Seal pups were repeatedly captured on a monthly basis from February through to July 2001. Pups were tagged at first capture and a blood sample was taken at each capture event. A total of 163 sera were collected from 118 seal pups. Where sufficient volume was collected, the sera were tested for leptospirosis using the microscopic agglutination test (MAT), and for brucellosis using the competitive enzyme-linked immunosorbent assay (ELISA) for Brucella abortus. RESULTS: None of 128 sera from 101 seals tested positive to the ELISA for B. abortus. All tests for Leptospira interrogans serovars Grippotyphosa, Copenhageni, Bratislava and Leptospira borgpetersenii serovar Ballum were negative at a cut-off of <1/100 dilution. Positive or suspicious titres were found to L. interrogans serovars Canicola and Pomona and L. borgpetersenii serovar Hardjo. The highest titres (12,800) were found to serovar Pomona. The titre to serovar Pomona in one seal rose from <1/50 in March to 12,800 in April and was <1/50 when re-sampled in July. The titre to serovar Pomona in another seal dropped from 12,800 in May to <1/50 in June. These seals also had titres to serovar Hardjo, which rose or fell in the same manner. All suspicious or positive titres occurred in late April and early May, when the pups were approximately 4-5 months old. In June and July, all seals tested were negative. CONCLUSIONS: There was no serological evidence of Brucella infection in the pre-weaned fur seals at the colony. Positive titres to serovars Pomona, Hardjo, or Canicola suggest that a Leptospira species was present at the colony, however isolation or visualisation of the organism is required to confirm this. Care should be exercised when handling New Zealand fur seals to prevent human infection or inadvertent transfer of leptospirosis to another marine mammal species. PMID- 16317445 TI - Effects of age on the electroencephalographic response to castration in lambs anaesthetised using halothane in oxygen. AB - AIM: To use the electroencephalogram (EEG) to ascertain whether the response of the cerebral cortex to the noxious stimulus of castration varied with age in lambs. METHODS: Two groups of East Friesian lambs were selected according to age; the mean age of the younger group (n=21) was 12 (SD 2) days and the older group (n=20) was 29 (SD 1) days. Anaesthesia was induced via mask using 4% halothane in oxygen, and maintained using 1.5% halothane in oxygen at a flow rate of 4 L/min. Once a stable plane of anaesthesia had been achieved, data collection of EEG and electrocardiographic (ECG) readings commenced, and the lambs were castrated 15 min later, using rubber rings. Median and 95% spectral edge frequencies (F95) and total EEG power (ptot) were derived from data from the EEG. RESULTS: Following castration, there was an increase in the median frequency (F50) in the younger lambs (p=0.002), and an increase in ptot in both groups (p=0.05), which was of greater magnitude in the older lambs. There were no significant changes in the F95. Both younger and older lambs exhibited a transient bradycardia (p=0.001 and p=0.01, respectively). CONCLUSIONS: These differences in the cortical response between the two groups suggest that 2-week-old lambs undergo a qualitatively different perception of the noxious stimulus of castration compared to 4-week-old lambs. PMID- 16317446 TI - Enlargements of the distal third metacarpus and metatarsus in Thoroughbred foals at pasture from birth to 160 days of age. AB - AIMS: To assess the relationship between the radiographic and microscopic appearance of the physeal regions of the distal third metacarpal (Mc3) and metatarsal (Mt3) bones of Thoroughbred foals at 160 days of age, and to clinically assess changes in contour of the distal Mc3 and Mt3 physeal regions from birth to 160 days of age. To assess relationships between maximum clinical physis scores and age, time of year, foal's sex, condition score, growth rate, copper (Cu) concentration in the liver of the foal, and supplementation of the dam with Cu in late gestation. METHODS: Dams were given Cu (n=5 and n=11, in Years 1 and 2, respectively) or saline (n=5 and n=12, in Years 1 and 2, respectively) injections in late gestation. Liver biopsies were harvested from foals in the first week of life, and the whole liver homogenised after the animals were euthanised at around 160 days of age, to determine Cu concentrations. Pasture samples were collected every 4-8 weeks for analysis of mineral composition. During Year 1, 10 foals were weighed and examined every 2 weeks from birth to 160 days of age for evidence of pain and lameness in the distal Mc3/Mt3. In Year 2, 23 foals were weighed, condition-scored and examined for evidence of pain and lameness weekly for the first 5 weeks of life, then every 2 weeks from birth to 160 days of age, and a clinical physis score for the distal Mc3/Mt3 given. Cabinet radiographs of frontal slices of the physeal region of the distal Mc3/Mt3 at around 160 days of age were given a radiographic physis score. Physes were then examined histologically for evidence of abnormal endochondral ossification. RESULTS: Gross enlargements of the distal Mc3 and Mt3 were observed in all foals in this study, but were not associated with lameness, pain or inflammation. The most severe clinical physis scores occurred over 2 months in late summer/autumn, and were not influenced by the foal's growth rate, sex, or Cu concentration in the liver, or treatment of dams with Cu in late gestation. The clinical physis score was highly correlated to radiographic evidence of shouldering in the forelimb and hindlimb (both p<0.001). Focal disturbances in endochondral ossification were evident radiographically and histologically in the some of the physes at 160 days of age. The mineral composition of pasture was similar in Years 1 and 2, and concentrations of Cu and zinc were below those currently recommended for growing horses. CONCLUSIONS: Gross enlargements of the distal Mc3 and Mt3 were not consistent with previous descriptions of physitis. Results suggest that while many Thoroughbred foals at pasture will have visible boney enlargements of the distal Mc3/Mt3 in the first 5 months of life, few have physeal cartilage abnormalities or significant compromise of endochondral ossification. The importance of these clinical swellings may be overestimated, and they may more appropriately be called physiological enlargements associated with remodelling of bone. PMID- 16317447 TI - Articular / epiphyseal osteochondrosis in Thoroughbred foals at 5 months of age: influences of growth of the foal and prenatal copper supplementation of the dam. AB - AIMS: To determine the influence of copper (Cu) supplementation by injection of mares in late gestation on the frequency and severity of osteochondrosis (OC) lesions in their foals at around 160 days of age. To determine if there was any influence of the concentration of Cu in the liver, growth rate, birthweight, weight at 160 days of age, fatness, sex, or year of birth of the foal on the frequency and severity of OC lesions. To determine the influence of dam's age, and sex and birthweight of the foal on the growth rate from birth to 160 days of age, and weight at 160 days of age. METHODS: Thirty-three Thoroughbred foals, born in two consecutive years, were weighed every 2 weeks from birth. The dams had been supplemented with parenteral Cu or saline during late gestation, and the supplementation regimens were different in each year. Foals had liver biopsies harvested at birth for determination of Cu concentration. Pasture samples were collected every 4-8 weeks for analysis of concentration of Cu and zinc (Zn). At 160 days of age, articular cartilage of long bones was examined. Gross lesions were counted and scored, then sawn and radiographed, and processed for histopathology. Lesions were given radiographic scores and histopathological scores. Maximum scores for each lesion were combined to give a total OC score for each joint and each foal. The fatness of 20 foals (10 each from Years 1 and 2) at 160 days of age was determined chemically. RESULTS: Supplementation of dams with Cu had no significant effect on the concentration of Cu in the liver of foals at birth, or on the frequency or severity of lesions in articular cartilage at 160 days of age. The Cu and Zn concentrations of pasture were similar in Years 1 and 2, and were lower than current recommendations. All foals in Year 2, and 9/10 foals in Year 1 had irregularities in cartilage that was confirmed histologically to be indicative of OC. The average number of lesions per foal was 4.7 (SD 1.1) and 5.7 (SD 1.1) in Years 1 and 2, respectively. However, the severity of the lesions was considered mild, and no foals showed any clinical evidence of OC while alive. The number of lesions in the tarsocrural (TC) joint and the TC OC score at 160 days were positively associated with average daily weight gain (ADG) in the previous 4 weeks (p=0.005 and p=0.001, respectively). There was no significant effect of sex, fatness, birthweight, weight at 160 days of age, or year of birth of the foal on the frequency and severity of OC lesions. CONCLUSIONS: Many of the lesions classified as OC, using classification systems described by other authors, were likely to be normal variations of the process of endochondral ossification. Despite the high frequency of such lesions, they were considered to be of minor significance and none were clinically evident. The distribution of lesions was not typical, and most probably reflected the subtlety of the lesions. These results support the hypothesis that Cu is an over emphasised factor in the aetiopathogenesis of OC. The relationship between subtle macroscopic lesions and lesions resulting in clinical signs of disease requires further investigation. PMID- 16317448 TI - Rapid detection and characterisation of infectious bronchitis virus (IBV) from New Zealand using RT-PCR and sequence analysis. AB - AIMS: To develop a reverse transcriptase-polymerase chain reaction (RT-PCR) assay to detect infectious bronchitis virus (IBV) from commercially-raised poultry in New Zealand and compare results with those from virus isolation. To characterise the IBV isolates using sequence analysis. METHODS: Pooled tissue samples (trachea, kidney, caecal tonsils and cloacal swabs) from 164 broiler and 53 layer flocks located throughout New Zealand were collected in transport medium containing antibiotics. Tissues were homogenised and the resultant supernatant used directly in a RT-PCR assay, and also inoculated into the allantoic cavity of 10-day-old embryonated eggs for virus isolation. Primers for the RT-PCR were selected from an area close to the N-terminus of the S1 (spike) gene and bracketed the hypervariable region 1 (HVR 1). The RT-PCR amplimers were sequenced from both termini, and alignment was constructed and analysed. RESULTS: From the 217 field samples that were subjected to RT-PCR, 42 (19%) were positive. Twenty nine (69%) of these RT-PCR-positive, and none of the RT-PCR-negative, samples yielded virus by isolation in chicken embryos. A phylogenetic tree constructed from these amplimers, that spanned the HVR of the S1 gene, revealed the IBV isolates clustered into two demarcated groups which had <60% homology. It is likely that the isolates of one group were derived from the live attenuated vaccine commonly used in New Zealand. CONCLUSIONS: The RT-PCR assay exhibited higher sensitivity than virus isolation and could be used for rapid diagnosis of IBV in the field. The prevalence of IBV appears to be surprisingly high in New Zealand although the use of pooled samples in the study did not allow accurate calculation of the prevalence in birds. Sequence analysis of a hypervariable region from the S1 gene was informative for the differentiation of closely related strains. PMID- 16317449 TI - Aspergillosis in a North Island robin (Petroica longipes). AB - CASE HISTORY: A 3-year-old female North Island robin (Petroica longipes) was found dead on Tiritiri Matangi Island during the breeding season. CLINICAL FINDINGS: The bird was in poor condition, and there was a 13 x 8 mm granulomatous mass in the thoracic cavity causing displacement of the heart and left lung. Histologically, the mass was a large granuloma infiltrated with fungal hyphae, and the liver contained multifocal aggregates of inflammatory cells. DIAGNOSIS: Thoracic aspergillosis and multifocal hepatitis. CLINICAL RELEVANCE: Determining the causes of death in populations of wild birds is often hampered by a lack of recovery of carcasses, autolysis and poor clinical history. In this case, the life history of the bird was known and recovery of the body was relatively swift. This is the first published description of aspergillosis in a free-living North Island robin. PMID- 16317450 TI - Ectopic hepatocellular carcinoma in a dog. AB - CASE HISTORY: A 14-year-old neutered male Bearded Collie was presented with a history of recurrent, intermittent urinary incontinence of 7 years duration. CLINICAL FINDINGS: A large, firm, non-painful mass was found in the mid-abdominal region on palpation. Ultrasonography of the mass revealed a compartmentalised structure with mixed echogenicity, and which did not appear to be associated with any of the abdominal organs. Ultrasound-guided fine needle aspirates contained several clusters of epithelial cells with cytological features of hepatocytes. At exploratory laparotomy, the mass was found in the gastrosplenic ligament within the greater omentum. PATHOLOGICAL FINDINGS AND DIAGNOSIS: Histopathologically, the mass consisted of sheets of hepatocytes, but without the characteristic hepatic architecture. The cells showed moderate variation in nuclear size and were sometimes binucleate. A diagnosis of hepatocellular carcinoma (HCC) in the mesentery was made. CLINICAL RELEVANCE: The presence of ectopic hepatic tissue has been reported rarely in man and cats, but not in the dog. Neoplastic transformation of ectopic hepatic tissue is seen in man. This is the first report of the presentation, clinical findings and treatment of a dog with ectopic HCC. PMID- 16317451 TI - Polymerase chain reaction (PCR) survey for rickettsias and bartonellas in ticks from New Zealand. PMID- 16317452 TI - Self-assembly and generation of complexity. AB - Supramolecular synthesis, by combining molecules through non-covalent interactions, makes it possible to build up extremely sophisticated supramolecular assemblies in the solid state and offers endless architectural possibilities. PMID- 16317453 TI - Engineering crystals by the strategy of molecular tectonics. AB - Detailed structures of molecular crystals cannot yet be predicted with consistent accuracy, but the strategy of molecular tectonics offers crystal engineers a powerful tool for designing molecules that are predisposed to form crystals with particular structural features and properties. PMID- 16317454 TI - Design of crystalline molecular networks with charge-assisted hydrogen bonds. AB - Empirical guidelines for steering molecular assembly into prescribed crystal architectures via hydrogen bonding continue to emerge, with recent developments demonstrating that charge-assisted hydrogen bonds introduce both strength and compliance that can facilitate solid state design. PMID- 16317455 TI - Structure-function correlations in oxygen activating non-heme iron enzymes. AB - A large group of mononuclear non-heme iron enzymes exist which activate dioxygen to catalyze key biochemical transformations, including many of medical, pharmaceutical and environmental significance. These enzymes utilize high-spin Fe(II) active sites and additional reducing equivalents from cofactors or substrates to react with O2 to yield iron-oxygen intermediates competent to transform substrate to product. While Fe(II) sites have been difficult to study due to the lack of dominant spectroscopic features, a spectroscopic methodology has been developed which allows the elucidation of the geometric and electronic structures of these active sites and provides molecular level insight into the mechanisms of catalysis. This review provides a summary of this methodology with emphasis on its application to the determination of important active site structure-function correlations in mononuclear non-heme iron enzymes. These studies provide key insight into the mechanisms of oxygen activation, active site features that contribute to differences in reactivity and, combined with theoretical calculations and model studies, the nature of oxygen intermediates active in catalysis. PMID- 16317456 TI - Solvent-induced supramolecular isomerism in [Pt(S=C(NH2)2)4]2+ croconate salts. AB - The role of solvent (and other species) in the formation of networks from tetrakis(thiourea)platinum(II) cations and croconate anions is examined, with crystallisations from DMSO giving rise to a structure containing solvent filled channels. PMID- 16317457 TI - A green to blue fluorescence switch of protonated 2-aminoanthracene upon inclusion in cucurbit[7]uril. AB - The inclusion of protonated 2-aminoanthracene in the cavity of cucurbit[7]uril increases its pKa values in the ground and excited states, resulting in the disappearance of the green emission of the neutral excited state and the significant enhancement of the blue emissions from the protonated excited state guest. PMID- 16317458 TI - Concomitant dimorphs of tri-O-[p-halobenzoyl]-myo-inositol 1,3,5-orthoformates with different halogen bonding contacts: first order crystal-to-crystal thermal phase transition of kinetic form to the thermodynamic form. AB - Crystallization of tri-O-[p-halobenzoyl]-myo-inositol 1,3,5-orthoformates from ethyl acetate-petroleum ether solution produced concomitant dimorphs that have different halogen bonding contacts; the kinetic form with C-Br...O-C contacts upon heating to 185 degrees C, converts completely to the thermodynamic form with C-Br...O=C contacts via crystal-to-crystal first order phase transition. PMID- 16317459 TI - Dependence of enzyme reaction mechanism on protonation state of titratable residues and QM level description: lactate dehydrogenase. AB - We have studied the dependence of the chemical reaction mechanism of L-lactate dehydrogenase (LDH) on the protonation state of titratable residues and on the level of the quantum mechanical (QM) description by means of hybrid quantum mechanical/molecular-mechanical (QM/MM) methods; this methodology has allowed clarification of the timing of the hydride transfer and proton transfer components that hitherto had not been possible to state definitively. PMID- 16317460 TI - Synthesis and properties of a stable 6-stannapentafulvene. AB - The first donor-free 6-stannapentafulvene stable at ambient temperature was synthesized and isolated, exhibiting the shortest tin-carbon bond length among those previously reported. PMID- 16317461 TI - Selective site controlled nucleophilic attacks in 5-membered ring phosphate esters: unusual C-O vs. common P-O bond cleavage. AB - Clean endocyclic C-O bond cleavage has been achieved in the reactions of 5 membered phosphate triesters with various nucleophiles. PMID- 16317462 TI - Collagen fiber immobilized Fe(III): a novel catalyst for photo-assisted degradation of dyes. AB - A novel catalyst for the Fenton reaction was prepared by immobilizing Fe(III) onto collagen fiber and its high activity as a catalyst for degradation of dyes under irradiation of UVC was proved. PMID- 16317463 TI - Reduction of alkynes into 1,2-dideuterioalkenes with hexamethyldisilane and deuterium oxide in the presence of a palladium catalyst. AB - A combination of hexamethyldisilane and deuterium oxide was found to work as a deuterium transfer reagent for alkynes in the presence of a catalytic amount of a palladium complex to give (E)-1,2-dideuterioalkenes selectively through the corresponding (Z)-isomer. PMID- 16317464 TI - Design, synthesis and evaluation of near-infrared fluorescent pH indicators in a physiologically relevant range. AB - Removal of a hydroxylsulfonylbutyl arm from indocyanine green dye produced a pH sensitive near-infrared (NIR) fluorescent indicator that is useful at physiological range. PMID- 16317465 TI - Reactivity patterns of thermally stable, terminal, electrophilic phosphinidene complexes towards diazoalkanes: oxidation at the phosphorus centre and formation of P-bound eta1-phosphaazine, eta1-phosphaalkene and eta3-diazaphosphaallene complexes. AB - The thermally stable, terminal phosphinidene complexes [CpM(CO)2(eta1 PNiPr2)]AlCl4(Cp= Cp, Cp*; M = Fe) and [Cp*M(CO)3(eta1-PNiPr2)]AlCl4 (M = Cr, Mo, W) react with Ph2C=N=N to form terminal P-coordinated eta1-phosphaazine and eta3 diazaphosphaallene ligands, respectively, whereas [CpFe(CO)2(eta1-PNiPr2)]AlCl4 reacts with Me3SiCHN2 affording a terminal phosphorus bound eta1-phosphaalkene complex. PMID- 16317466 TI - Reactions of P/S-containing proligands with coinage metal salts: a new route to polynuclear complexes with unusual structural types. AB - The reactions of P/S-precursors with coinage metal alkoxides and carboxylates offer new perspectives for the synthesis of complex metal aggregates. PMID- 16317467 TI - Porous carbon-carbon composite replicated from a natural fibre. AB - Highly porous carbon composites, suitable to be used as electrodes in electrochemical double layer capacitors, could be produced by pyrolysis of resorcinol-formaldehyde resins impregnated onto a natural fibre material (celullosic fabric), without additional template agents or special drying of the polymer gels. PMID- 16317468 TI - Cross-coupling of benzylic acetates with arylboronic acids: one-pot transformation of benzylic alcohols to diarylmethanes. AB - Benzylic acetates reacted with arylboronic acids in the presence of a DPEphos [Pd(eta3-C3H5)Cl]2 catalyst when tert-amyl alcohol was used as a solvent, and the catalytic cross-couplings produced diarylmethanes in high yields (up to 94% isolated yield). PMID- 16317469 TI - The solvated fluoride anion can be a good nucleophile. AB - Computations show how the solvated fluoride ion can be a good nucleophile in spite of its high solvation energy. PMID- 16317470 TI - Latent reactive groups unveiled through equilibrium dynamics and exemplified in crosslinking during film formation from aqueous polymer colloids. AB - The concept of using equilibrium dynamics to provide for both protection and unveiling of latent functional groups at appropriate times in aqueous polymer colloid coatings designed for crosslinking only during film formation is introduced; the new functional monomer, 4-hydroxyethylsulfonylstyrene (HESS), readily undergoes emulsion copolymerization with acrylates to form stable latexes, followed by crosslinking by loss of water during film formation. PMID- 16317471 TI - Rod-like Cu/La/O nanoparticles as a catalyst for phenol hydroxylation. AB - Rod-like La/Cu/O nanoparticles, synthesized by a simple coprecipitation reaction with a sonication process, are found to be highly active as a catalyst for the hydroxylation of phenol. Compared to the 4-6 h, 40% yield reported in the literature, our nanoparticle catalyst demonstrates a nearly 100% conversion within 4 h based on gas chromatography. PMID- 16317472 TI - Controlling the speed of rotation in molecular motors. Dramatic acceleration of the rotary motion by structural modification. AB - Substitution of a 6-membered by a 5-membered ring upper half in the light driven second generation molecular motors resulted in a dramatic increase of the speed of rotation. PMID- 16317473 TI - Enzyme responsive polymer hydrogel beads. AB - We report on a new class of enzyme responsive polymer hydrogels, the molecular accessibility of which can be changed selectively by enzymes present in a sample fluid. PMID- 16317474 TI - Inactivation of human angiotensin converting enzyme by copper peptide complexes containing ATCUN motifs. AB - The copper complex [KGHK-Cu]+ demonstrates catalytic inactivation of human angiotensin converting enzyme at sub-saturating concentrations, under oxidative conditions, with an observed rate constant k approximately 2.9 +/- 0.5 x 10(-2) min(-1). PMID- 16317475 TI - Hydrogen evolution from organic "hydrides". AB - Benzimidazolines (dihydrobenzimidazoles) are shown for the first time to eliminate hydrogen (H2) by catalyzed reaction with protic compounds. PMID- 16317476 TI - [LnIIAlIII2(alkyl)8]x: donor addition instead of donor-induced cleavage. AB - (SmAl2Me8)x and (SmAl2Et8)x are obtained via a silylamide elimination reaction from Sm[N(SiMe3)2]2(THF)2 and excess AlR3 (R = Me, Et); (LnAl2Et8)x (Ln = Sm, Yb) react with THF, pyridine, and 1,10-phenanthroline to form the first donor adducts of homoleptic peralkylated Ln-Al heterobimetallic complexes. PMID- 16317477 TI - Samarium versus aluminium Lewis acidity in a mixed alkyl carboxylate complex related to alkylaluminium activation in diene polymerization catalysis. AB - [(C5Me5)2Sm(mu-O2CPh)]2 reacts with iBu3Al to form a mixed bridge samarium aluminium complex [(C5Me5)2Sm(mu-O2CPh)(mu-iBu)Al(iBu)2], that displays two different carboxylate orientations toward the metals in a single crystal. PMID- 16317478 TI - Generation and ring opening of aziridine N-carbonyl radicals. AB - Aziridine N-carbonyl radicals, generated by irradiating the corresponding S oxalyl xanthates, undergo ring opening to give 2-isocyanato radicals, which can be trapped by an external olefin. PMID- 16317479 TI - [Spinal anaesthesia]. AB - In spinal anaesthesia, surgical analgesia and in most cases motor block is achieved by injecting one or more drugs into the cerebrospinal fluid. As one of the earliest methods of anaesthesia it was introduced into clinical practice in the late nineteenth century. Although later on it was more or less replaced by "modern" general anaesthesia, it has regained popularity due to its benefits for certain patient populations. In spite of being a technically simple procedure, performing spinal anaesthesia requires a sound knowledge of applied physiology and pharmacology, especially in high-risk groups such as pregnant women or former preterm babies. For some patients even under anticoagulant therapy, spinal anaesthesia might be the best option, considering the individual risk of alternative methods. PMID- 16317480 TI - Morphology and productivity of filamentous fungi. AB - Cultivation processes involving filamentous fungi have been optimised for decades to obtain high product yields. Several bulk chemicals like citric acid and penicillin are produced this way. A simple adaptation of cultivation parameters for new production processes is not possible though. Models explaining the correlation between process-dependent growth behaviour and productivity are therefore necessary to prevent long-lasting empiric test series. Yet, filamentous growth consists of a complex microscopic differentiation process from conidia to hyphae resulting in various macroscopically visible appearances. Early approaches to model this morphologic development are recapitulated in this review to explain current trends in this area of research. Tailoring morphology by adjusting process parameters is one side of the coin, but an ideal morphology has not even been found. This article reviews several reasons for this fact starting with nutrient supply in a fungal culture and presents recent advances in the investigation of fungal metabolism. It illustrates the challenge to unfold the relationship between morphology and productivity. PMID- 16317481 TI - Uptake and persistence of the vertebrate pesticide, sodium monofluoroacetate (compound 1080), in plants of cultural importance. AB - Field research was undertaken to determine if naturally occurring plants utilised by a Maori (the indigenous people of New Zealand) community for food and medicine would take up the toxin sodium monofluoroacetate (Compound 1080) from baits used to control the brush-tailed possum Trichosurus vulpecula. Single baits were placed at the base of individual plants of two species, pikopiko (Asplenium bulbiferum) and karamuramu (Coprosma robusta). Plants were sampled at various times up to 56 days, and samples analyzed for 1080 content. No 1080 was detected in any of the pikopiko samples, whereas 1080 was detected in karamuramu, at a maximum concentration of 5 ppb after 7 days, and 2.5 ppb after 14 days. This concentration decreased to zero at 28 days, indicating that while karamuramu was shown to take up 1080, it was not persistent. The results of this study suggest there is negligible risk of humans being poisoned by consuming plants that have taken up 1080 from baits. To allay community concerns that minute concentrations of 1080 might influence the medicinal properties of plants, it is suggested a withholding period of 30 days after 1080 control operations could be adopted. PMID- 16317482 TI - Site specific PCB-correlated interspecies differences in organ somatic indices. AB - We correlated site specific differences in the organ somatic indices of nestlings of five passerine species (tree swallow, red-winged blackbird, house wren, Carolina chickadee, and eastern bluebird) with the degree of polychlorinated biphenyls (PCB) exposure in ovo and post-hatching. The birds were exposed to PCBs at or downstream of four PCB-contaminated sites. Of the organs evaluated for this paper, brain, bursa, heart, kidneys, lungs, pancreas, spleen, stomach, and thyroid varied significantly (p<0.05) or marginally significantly (0.05or=3 mm was observed in 47 patients (16%) and 100 microM) decrease level of GSH was noticed. Increased levels of metallothionein in PBMCs were observed in response to zinc. DNA fragmentation studies also showed that copper produced DNA fragmentation at LD(50) (115 microM). Subsequently, zinc showed protection against DNA fragmentation caused by copper. Cell structure of PBMCs at LD(50) (115 microM copper) showed membrane bound cystic spaces and mitochondria having disrupted cristae and few myelin figures. In presence of zinc at LD(50) of copper (115 microM) cells showed improvement in mitochondrial structure and membrane bound cystic spaces. Taken together, the results of our study demonstrates that zinc play an important role in prevention of copper toxicity in peripheral blood mononuclear cells. PMID- 16317508 TI - Lupeol and its ester ameliorate the cyclophosphamide provoked cardiac lysosomal damage studied in rat. AB - Cyclophosphamide (CP), an alkylating agent widely used in cancer chemotherapy causes fatal cardiotoxicity. Lupeol, a pentacyclic triterpene, isolated from Crataeva nurvala stem bark and its ester, lupeol linoleate possess a wide range of medicinal properties. The effect of lupeol and its ester was evaluated in CP induced myocardial toxicity in rats. Male albino rats of Wistar strain were categorized into six groups. Group I served as control. Rats in groups II, V and VI animals were injected intraperitoneally with a single dose of CP (200 mg/kg body weight) dissolved in saline. CP-treated groups V and VI received lupeol and lupeol linoleate (50 mg/kg body weight), respectively, dissolved in olive oil for 10 days by oral gavage. CP-administered rats showed a significant increase (p < 0.001) in the activities of lysosomal hydrolases in serum and heart, a decrease (p < 0.001) in the levels of cellular thiols and myofibres were swollen with loss of myofilaments in electron microscopical analysis in heart. Lupeol and its ester showed reversal of the above alterations induced by CP. These findings demonstrate that the supplementation with lupeol and its ester could preserve lysosomal integrity, improve thiol levels, highlighting their protective effect against CP-induced cardiotoxicity. PMID- 16317509 TI - Prevention of doxorubicin-induced cardiotoxicity by melatonin. AB - Anthracyclines, such as doxorubicin and daunorubicin, are highly effective anticancer agents. Cardiotoxicity made by these agents develops as a complication of the cancer chemotherapy. Melatonin, the chief secretory product of the pineal gland, was recently found to be a free radical scavenger and antioxidant. We decided to evaluate the tissue-protective effect of melatonin against myocardial toxic effects of doxorubicin in six groups of rats. Rats were given doxorubicin (Dx) (45 mg/kg dose) and melatonin (MEL) (10 mg/kg), first doxorubicin and then melatonin (DM), first melatonin and then doxorubicin (MD). The degree of cardiac muscle cell alterations were examined either histologically (mean total score technique) or biochemically. In doxorubicin-treated group, malondialdehyde (MDA) levels of the heart tissue were significantly increased, glutathione (GSH) levels were decreased compared to the control rats. In the group in which first doxorubicin and then melatonin was given, MDA levels were significantly decreased and glutathione (GSH) levels were increased compared to the doxorubicin-treated group. During ultrastructural analysis, in doxorubicin-treated group, cellular edema, mitochondrial deformation, decreased glycogen stores, and disordered myofibrillary structure were observed. In contrast, in first doxorubicin and then melatonin-treated group, normal cellular structure was observed. But, first melatonin and then doxorubicin-treated group was not significantly preserved from the doxorubicin-induced changes. By preventing lipid peroxidation and myocardial lesions, melatonin may be highly effective in protecting against doxorubicin induced cardiotoxicity. PMID- 16317510 TI - Nucleotides induce higher order chromatin degradation. AB - Higher order chromatin degradation (HOCD) is a stepwise dismantling of the genome through the excision of chromatin loops and their oligomers at matrix attachment regions (MARs) during the early stages of programmed cell death. Although HOCD ultimately leads to the inactivation of the genome and cell death, a partial HOCD in cells receiving sublethal signals may result in the loss of genetic stability leading to neoplasia, degeneration, and aging. The present study was undertaken to determine the role of protein poly(ADP-ribosyl)ation in HOCD. Nuclei isolated from rat glioma C6 cells were able to carry poly(ADP-ribosyl)ation as assessed by the incorporation of (32)P-NAD(+) into TCA-insoluble fraction. Under the same experimental conditions, millimolar NAD(+) induced rapid HOCD in nuclei. However, while poly(ADP-ribosyl)ation was totally abrogated by specific inhibitor, benzamide, NAD(+)-induced HOCD was unaffected. Benzamide also failed to inhibit HOCD induced by H(2)O(2) exposure in intact cells. These results indicate that HOCD is not mediated through chromatin poly(ADP-ribosyl)ation, and that NAD(+) activates MAR-associated endonuclease or facilitates the access of the enzyme to DNA by other mechanisms. Furthermore, other nucleotides including NADP(+), ATP, UTP, GTP, and CTP were also found to induce HOCD in isolated nuclei indicating that HOCD is controlled by nucleotide-related ligands. PMID- 16317511 TI - Alcohol-induced bone degradation and its early detection in the alcohol-fed castrated rats. AB - The objective of this study was to examine alcohol-induced changes of bone in hormone-deficient males using the developed method. In the process of bone resorption, type I collagen crosslinking molecules, pyridinoline (PYD), are released into the circulation and cleared by the kidneys. (2)H(2)O as a tracer has been applied to measure the synthesis rates of slow-turnover proteins and successfully applied to bone collagen synthesis in our hormone deficiency rats. This study demonstrated for the first time, the early changes of the femur bone degradation in hormone-deficient male individuals, more influenced by alcohol through histopathological study, serum PYD assay, and (2)H(2)O labeling. We also observed that serum PYD was a sensitive pathological marker of bone degradation in castrated osteoporosis males and the unique features of (2)H(2)O labeling to measure the bone turnover collagen synthesis rates were excellent markers of bone degradation and aging. PMID- 16317512 TI - Reduced sarcoplasmic reticulum Ca2+ -ATPase activity and dephosphorylated phospholamban contribute to contractile dysfunction in human hibernating myocardium. AB - Human hibernating myocardium (HHM) is characterized by reversible contractile dysfunction during chronic ischemia. A disturbed calcium-homeostasis is a decisive factor for reduced functional capacity in heart diseases. We therefore investigated calcium-handling proteins in HHM. In 12 patients suffering from multi-vessel coronary artery disease and contractile dysfunction with indication for bypass surgery, HHM was detected preoperatively by thallium scintigraphy, radionuclide ventriculography and dobutamine echocardiography. Transmural biopsies of these regions were taken and analyzed by immunohistochemistry and electron microscopy. Furthermore, SR-calcium ATPase (SERCA2a), phospholamban (PLN), the phosphorylated forms of PLN (PLN-Ser16, PLN-Thr17) as well as sodium calcium exchanger (NCX) and ryanodine receptor (RyR2) were investigated by RT-PCR and Western-blotting. Additionally, SERCA2a activity was measured by an enzyme coupled assay. In all patients complete functional recovery could be documented 3 months after revascularization by repeating all preoperative investigations. In HHM maximal SERCA2a activity was significantly reduced (HHM: 424.5 +/- 33.9, control: 609.0 +/- 48.5 nmol ATP mg protein(-1) min(-1), p